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III ABCF Congress | Frontiers of Pharmaceutical Sciences in the Omics Era
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COMMITTEES
ABCF DIRECTORY BOARD
President: João Carlos Palazzo de Mello (UEM)
Vice-President: Teresa Dalla Costa (UFRGS)
Secretary: Lúcio Mendes Cabral (UFRJ)
First Secretary: Flávio da Silva Emery (USP-RP)
Treasurer: Leonardo Régis Leira Pereira (USP-RP)
First Treasurer: Armando da Silva Cunha Júnior (UFMG)
ORGANIZING COMMITTEE
Teresa Dalla Costa (UFRGS) – President
Helder Teixeira (UFRGS) – Secretary
Mauro Silveira de Castro (UFRGS) – Treasurer
Alexandre Macedo (UFRGS)
Clarice Rolim (UFSM)
Clésio Soldateli Paim (UNIPAMPA)
Gilsane Lino Von Poser (UFRGS)
Tiana Tasca (UFRGS)
SCIENTIFIC COMMITTEE
Renata Vianna Lopez (USP/RP) - President
Celso Nakamura (UEM)
Claudia Simões (UFSC)
Isabela Heineck (UFRGS)
Lídia Moreira Lima (UFRJ)
Maira Galdino da Rocha Pitta (UFPE)
Norberto Lopes (USP/RP)
Raquel Giordani (UFRN)
Sâmia Joca (USP/RP)
Stela Maris Kuze Rates (UFRGS)
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POSTER EVALUATING COMMITTEE
Clarice Rolim (UFSM) – Coordinator
Clésio Soldateli Paim (UNIPAMPA) - Coordinator
Adriana Raffin Pohlmann (UFRGS)
Ana Luisa Palhares de Miranda (UFRJ)
Ana Maria Bergold (UFRGS)
Andrea Diniz (UEM)
Andréa Inês Horn Adams (UFSM)
Andreas Loureiro Mendez (UFRGS)
Angela Machado de Campos (UFSC)
Angélica Garcia Couto (UNIVALI)
Bibiana Verlindo de Araújo (UFRGS)
Carlos Alexandre Carollo (UFMS)
Carolina de Miranda Silva (UFRGS)
Clesio Soldateli Paim (UNIPAMPA)
Cristiana Lima Dora (FURGS)
Cristiane de Bona Da Silva (UFMS)
Cristiane dos Santos Giuberti (UFES)
Cristiane Franco Codevilla (UFSM)
Denise Brentan (UFMS)
Diogo Pilger (UFRGS)
Divaldo Lyra Junior (UFSE)
Eduardo Luis Konrath (UFRGS)
Eliana Elisabeth Diehl (UFSC)
Fávero Reisdorfer Paula (UNIPAMPA)
Fernão Castro Braga (UFMG)
Francine Johansson Azeredo (UFBA)
Gabriela Garrastazu Pereira (UFRGS)
Gabriele Dadalt Souto (UFRGS)
Humberto Gomes Ferraz (USP)
Juliana Bidone (UFRGS)
Karina Paese (UFRGS)
Leandro Tasso (UCS)
Maiara Cássia Pigatto (UFRGS)
Marcelo Donadel Malesuik (UNIPAMPA)
Marco Vinícius Chaud (UNISO)
Maria Palmira Daflon Gremião (UNESP)
Miriam Apel (UFRGS)
Moacyr Jesus Barreto de Melo Rêgo (UFPE)
Mônica Cristina Oliveira (UFMG)
Nelilma Romero (UFRJ)
Roberto Pontarolo (UFPR)
Rodrigo José Freddo (UNIPAMPA)
Sandra Elisa Haas (UNIPAMPA)
Saulo Fernandes de Andrade (UFRGS)
Silvya Stuchi Maria-Engler (USP)
Simone Cristina Baggio Gnoato (UFRGS)
Tadeu Uggere de Andrade (UVV)
Tânia Alves Amador (UFRGS)
Thiago Caon (UFSC)
Tiana Tasca (UFRGS)
Vitor Todeschini (UNIPAMPA)
ABSCTRACT EVALUATING COMMITTEE
Gilsane Lino von Poser (UFRGS) – Coordinator
Analytical methods for drugs and medicines
Celso Nakamura (UEM)
Andréia Adams (UFSM)
Cássia Garcia (UFRGS)
Cleber Schmidt (UFBA)
Vítor Todeschini (UFRJ/Macaé)
Keyller Borges (UFSJ)
Marcelo Malesuik (UNIPAMPA)
Martin Steppe (UFRGS)
Simone Cardoso (UFSC)
Biological Assays, Biomarkers and Diagnostics
Maira Pitta (UFPE)
Tiana Tasca (UFRGS)
Silvia Berlanga (USP-SP)
Moacyr Jesus Barreto de Melo Rêgo (UFPE)
Silvya Stuchi Maria-Engler (USP/SP)
Drug discovery, Medicinal Chemistry and
Molecular Medicine
Daniel Kawano (UNICAMP)
Fávero Paula (UNIPAMPA)
Flávio Emery (USP/RP)
Marcelo Zaldini Hernandes (UFPE)
Giuliano Cesar Clososki (USP/RP)
Lídia Moreira Lima (UFRJ)
Natural Products and Toxinology
Eduardo Konrath (UFRGS)
Fernão Braga (UFMG)
Norberto Peporini Lopes (USP/RP)
Rachel Castilho (UFMG)
Raquel Giordani (UFRN)
Tania Bresolin (UNIVALI)
Pharmaceutical and cosmetic technology
Adriano Antunes de Souza Araújo (UFS)
Airton Monza da Silva (PUCRS)
Angela Campos (UFSC)
Cristiane De Bona da Silva (UFSM)
Edison Carvalho (UFRJ/Macaé)
Elenara Lemos-Senna (UFSC)
Fernanda Bruxel (UNIPAMPA)
Kellen Borges (UFSCPA)
Letícia Colomé (UNIPAMPA)
Leticia Cruz (UFSM)
Letícia Köester (UFRGS)
Maria Ismenia Lionzo (UFSCPA)
Renata Vianna (USP-RP)
Scheila Schaffazick (UFSM)
Sandra Haas (UNIPAMPA)
Pharmaceutical services
Diogo Pilger (UFRGS)
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Divaldo Pereira de Lyra Júnior (UFS)
Isabela Heineck (UFRGS)
Leonardo Régis Leira Pereira (USP-RP)
Tadeu Uggere de Andrade (UVV – Vila Velha)
Tatiane da Silva Dal Pizzol (UFRGS)
Pharmacology and Toxicology
Claudia Simões (UFSC)
Marilu Fiegenbaum (UFSCPA)
Sâmia Joca (USP)
Stela Rates (UFRGS)
Andreia Diniz (UEM)
Francine Azeredo (UFBA)
ABCF YOUNG INVESTIGATOR AWARD 2016
Committee
Tania Mari Bellé Bresolin – President
Letícia Cruz
Maira Galdino da Rocha Pitta
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Afranio Lineu Kristi
Federal University of Rio de Janeiro
Brazil
NEGLECTED AND EMERGING INFECTIOUS DISEASES
The “Global End TB Strategy” aims to reduce TB deaths and incidence in all countries, to levels that are now
observed in high-income countries. The strategy is based in three pillars: Pillar 1 (integrated, patient-centered care
and prevention), Pillar 2 (bold policies and supportive systems), and Pillar 3 (intensified research and innovation).
Brazil as one of few “model” countries that already have substantial TB research capacity and could achieve these
goals rapidly, and the existing Brazilian TB Research Network (Rede TB), working closely with the National TB
Program (NTP), Ministry of Health (MoH) in the last 10 years will be instrumental to the development of the
strategy in the country. The need to develop a National TB Research Agenda through different Research Platforms
that could cover several disciplines and include it in the National TB Research Strategy Plan was a consensus
reached in several meetings organized by Rede TB. Working groups organized around key TB research activities
identified research gaps and research priorities. These Research Platforms were consolidated as key steps in
developing and implementing a National TB Research Strategic Plan that will be presented and discussed with
several national and international stakeholders. It will be presented at the Brazilian Association of Pharmaceutical
Sciences (ABCF) how can be developed in Brazil the Pillar 3 of the new “End TB Strategy” proposed by WHO and
integrate it with the other two Pillars. This agenda will be an integral part of the National Tuberculosis Research
Strategy Plan to be launched in 2016.
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Bernd Meibohm
University of Tennessee
USA
PK/PD MODELING OF THERAPEUTIC PROTEINS
Therapeutic proteins, particularly monoclonal antibodies are the largest and most rapidly growing class of
therapeutics and are at the forefront of many efforts related to precision medicine. The success of protein-based
therapeutics can be attributed to the numerous advantages that this class of biologics carries including the ability
to create therapeutics with high affinity and selectivity for a wide range of targets, the potential to modulate the
native properties of proteins for customized therapeutic applications and the usually high tolerability and limited off
target toxicity. Despite their many favorable properties, however, there are also multiple challenges associated
with the development of proteins as therapeutics. Their large size hinders tissue penetration and delivery to
intracellular targets. Immunogenicity has led for numerous therapeutic proteins to the formation of anti-drug
antibodies which was subsequently responsible for a reduced systemic exposure, loss of efficacy, and/or adverse
events. Recent advances in protein engineering have opened up new avenues of modulating the desired
properties of therapeutic proteins. This includes not only the optimization of existing proteins by approaches such
as glycoengineering, but also the creation of new scaffolds such as antibody-drug conjugates (ADCs), antibody
fragments, antibody-based fusion proteins, nanobodies, and bispecifics. The presentation will focus on how
pharmacometrics and quantitative pharmacology approaches can provide a scientific framework that facilitates,
supports, and ultimately accelerates the successful development of therapeutic proteins, particularly through
PK/PD-based modeling and simulation approaches.
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Bibiana Verlindo de Araújo
Federal University of Rio Grande do Sul
Brazil
HOW THE PK-PD MODELING IN PRE-CLINICAL STUDIES CAN HELP US TO BETTER
UNDERSTAND HOST-MICROORGANISM-DRUG RELATIONSHIP?
In spite of the improvement in drug therapy observed in the last decades, infections are still a great problem that
affects the whole world in different degrees, independent of social or economic conditions. Good results after
antimicrobial therapy depends on the triad: microorganism-host-drug and as a consequence it is crucial to analyze
the connections between these key factors. Recognizing the importance of studies linked to further comprehension
of this interaction and its impact on the drug efficacy, our research group has been working with experimental
models of infection, investigating their influences on tissue drug distribution by microdialysis and connecting the
results by pharmacokinetics-pharmacodynamics modeling. The aim is to clarify some aspects related to the failure
in antimicrobial therapy observed in clinical scenarios and propose new directions. Three cases will be discussed
in this lecture: a) differences observed in the PK/PD parameters for the same drug and microorganism when were
used in vitro or in vivo models to predict the effect of piperacillin against infections caused by Escherichia coli; b)
the impact of infection on antifungal’s distribution in the brain after experimental disseminated cryptococcosis in
rats and c) how the population pharmacokinetics can be applied in pre-clinical studies allowing a better
understanding of host-microorganism-drug.
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Carlos Alexandre Carollo
Federal University of Mato Grosso do Sul
Brazil
METABOLOMICS AS A TOOL IN NATURAL PRODUCTS RESEARCH
Natural products stand out in the search for new drugs leads, mainly due to the abundant structural variety of
compounds. This search occurs for many years in the classic way, which involves extractions and chromate
graphic separation, allowing only the isolation of the majority compounds. Despite the success along the years
decreased the number of new molecules discovered and was necessary the inclusion of new rational analysis
tools. Since then, we observed an increasing number of articles that use metabolomics to accelerate the discovery
of new compounds, using hyphenated techniques as HPLC-MSand statistical approach. This strategy avoid the
isolation of known compounds, allow the identification of patterns of distribution of secondary metabolites, assists
in the understanding of species evolution and in the identification of compounds responsible for biological activity
in plant extracts without necessity of isolation and re-evaluations.
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Cesar Augusto Souza Andrade
SECTEC/PE, Federal University of Pernambuco,
Brazil
PARTNERSHIPS FOR THE PRODUCTIVE DEVELOPMENT: GOVERNMENT ACTIONS TO
REDUCE NATIONAL DEPENDENCE ON HEALTH PRODUCTS
Brazilian govern mental unched a set of policies to improve the competitivit yof nation al industry and reduce the
dependence o nimported medicines and medical devices. Of note, Partnerships for the Productive Development
(PDPs) are a result of govern mentef forts to expand domestic production through agree ments between publicand
private firms. The Ministry of Health by means of PDP saim sexpand the local production of medicines and medical
devices considered strategicto the Brazilian National Health Service (SUS). Therefore, PDPs are analternative to
reduce prices of innovative drugsand medical devices, guarantee patientaccess to innovative treatments, and
promote savings to the system. In spite of Brazilian internal difficulties and recession, the future commercial
outlook of the Brazilian domestic industry looks positive. In addition, international pharma companies have seen
the PDPs as a use fulopp or tunity to consolidate their position in the Brazilian Market. The increase in the
production capacity of the public laboratories may negatively affect the international pharmaceutical industry. Thus,
in this talk we will discuss the effectivity of PDPsto improve the competitivey of the public laboratories to produce
medical device sand medicines.
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Chantra Eskes
SeCAM Services & Consultation for Alternative
Methods, Switzerland
CURRENT ACCEPTANCE AND FUTURE PROSPECTS ON THE USE OF ALTERNATIVE
TEST METHODS FOR TOXICITY TESTING
The last years have been marked by the international acceptance of a number of in vitro alternative methods to
animal testing adopted by the OECD program on Test Guidelines. In order for alternative methods to gain such
international regulatory acceptance, they traditionally need to undergo a process of independent scientific
validation.
A number of in vitro test methods have been validated and adopted in the last decade, in particular in the area of
topical toxicity hazard assessment. In some cases it is possible to fully replace the animal test, such as for skin
irritation and corrosion by using Integrated Approaches for Testing and Assessment (IATAs). In other cases, it is
possible to partially replace the animal test such as for eye hazard and skin sensitization identification. In particular
for skin sensitization, Defined Approaches that combine e.g., in chemico and in vitro test methods will most likely
be needed to address the various key events of the Adverse Outcome Pathway (AOPs) leading to allergic contact
dermatitis in humans or contact hypersensitivity in rodents.
The use of these new approaches for safety assessment such as the IATAs and the AOPs, bring a number of
questions related to the application of alternative methods such as their possible combination in testing strategies
and/or defined approaches. The latest progress and new challenges facing the application of alternative test
methods for regulatory safety assessment will be presented.
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Denise Brentan da Silva
Federal University of Mato Grosso do Sul,
Brazil
MALDI MASS SPECTROMETRY: FROM METABOLOMICS TO SPATIAL LOCALIZATION
OF METABOLITES IN TISSUE
MALDI-MS has been applied to identify the chemical structures, metabolomic studies, quantification and, recently,
to analyze the distribution of metabolites within tissues. MALDI imaging mass spectrometry combines the
molecular mass analysis and spatial, which was firstly used to analyze animal tissue but currently includes other
tissue analyses, such as plant, microorganism, insect and others. MALDI imaging, unlikely classical histochemical
methods (unspecific), can distinguish between individual compounds and confirm it by MS/MS data and high
accuracy, which both reduce the problems related to isobaric matrix ions and create images data more reliable.
From new technologies overcoming the bottlenecks of MALDI analyses, it is possible have access to all its benefits
enlarging its application in different areas, mainly because of MALDI technique exhibits several advantages, such
as high sensitivity, high ability to analyze complex mixtures with less ion suppression compared to ESI, high
tolerance of salts and contaminants, high throughput, low sample consumption, low time consumption to obtain the
spectra (≈ 60 s).
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Fabrício de Araujo Moreira
Federal University of Minas Gerais,
Brazil
CANNABIDIOL AS A MULTI-TARGET DRUG FOR TREATING NEURO-PSYCHIATRIC
DISORDERS
Cannabidiol (CBD) is a compound from Cannabis sativa, an herb widely consumed as a drug of abuse under the
names of marijuana, hemp or hashish. Delta-9- tetrahydrocannabinol (THC) is the main compound responsible for
cannabis psychoactive effects. Although both compounds are interesting from a therapeutic standpoint, THC has
addictive, memory-impairing and psycho tomimetics properties that limits its applications. In this context, CBD has
emerged as a major target for pharmacological investigation. This compound is devoid of the deleterious effects of
THC and may have therapeutic efficacy in a various neurological and psychiatric disorders.
The pioneering studies on the chemistry and pharmacology of CBD were performed by Prof. Raphael Mechoulam
in Israel, around 50 years ago. Since then, various Brazilian scientists have also played a major role in
investigating this compound.
Most of these studies focused on experimental models in laboratory animals. More recently, human studies have
also been performed in healthy volunteers and patients.
In this context, the aim of this talk is to provide a short historical background related to the pharmacology of CBD
and to present recent original data focusing on its therapeutic potential for brain disorders, including schizophrenia,
anxiety and epilepsy.
As it will be discussed, decades of research suggest that CBD is a promising compound that may lead to new
pharmaceuticals with higher efficacy and more favourable side effect profiles for the treatment of neuro-psychiatric
disorders.
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Fernando Batista da Costa
University of São Paulo,
Brazil
PLANT METABOLOMICS IN PHARMACEUTICAL SCIENCES
Metabolomics is a discipline dedicated to the study of low--molecular--weight metabolites of organisms or
biological matrices by comprehensive and sensitive analytical techniques. The analytical techniques that are most
often used for this purpose are nuclear magnetic resonance (NMR), high--performance liquid chromatography
coupled with mass spectrometry (HPLC--MS) and gas chromatography coupled with mass spectrometry
(GC--MS). Metabolomics is combined with suitable multivariate statistical analysis and in silico tools to classify
the samples into different groups or to determine which compounds are correlated with the investigated property or
characteristic of the sample.
In Pharmaceutical Sciences, plant metabolomics may have different applications, such as quality control of herbs,
determination of geographic origin and discovery of biomarkers. The Aster BioChem research group has been
studying species from the sunflower family (Asteraceae) for several years and recently incorporated plant
metabolomics. A pure compound library, an extract library and an in house database have been developed to
support extract dereplication. Therefore, an interesting platform was established and made possible to focus on
plant metabolites. In this presentation, two examples will be presented. In the first, several plant extracts have
been systematically investigated by LC--MS and the information concerning the direct inhibition of the enzymes
cyclooxygenase--1 (COX--1) and 5--lipoxygenase (5--LOX) using these extracts was obtained. Then, the
extracts with previously available information from LC--MS analysis and in vitro dual inhibition of COX--1
and 5--LOX were selected for in silico studies using machine learning algorithms. The chromatographic peaks
of the extracts were extracted, processed, selected and analyzed by in silico methods. As a result, 11 compounds
were determined to be biomarkers due to their anti--inflammatory potential. In the second example, the influence of
different abiotic environmental factors (climate and soil) on plant metabolites has been investigated by LC--MS and
1H NMR. Samples of leaves, stems, roots and inflorescences of the medicinal plant Tithonia diversifolia were
collected along with the corresponding monthly environmental data, throughout a 24--month period. A
metabolomic approach was proposed to determine the relationship between environmental changes and plant
metabolism. The results obtained from both analytical techniques were complementary in this work. A seasonal
pattern was observed for the leaf and stem metabolites, which correlated to the climate data. The inflorescence
and root metabolites were mainly affected by the presence of some soil nutrients. We therefore conclude that plant
metabolomics is an important field of research with interesting applications in Pharmaceutical Sciences. Parasites
epigenome rises as an interesting target to drug discovery programs. Epigenetics comprises a series of chemical
modifications of DNA and their associated histone proteins and it is known to be an especially important aspect of
parasite biology, although it is underexplored in drug discovery programs. In this talk, I will cover the main aspects
of epigenetic drug discovery strategies for neglected diseases, focusing on compounds design and library
development and characterization. I will be focusing in parasitic Sirtuin family of enzymes to provide examples of
the Laboratory of Heterocyclic and Medicinal Chemistry - QHETEM.
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Flavio Guimaraes da Fonseca
Federal University of Minas Gerais,
Brazil
ALTERNATIVE TECHNOLOGIES IN THE DEVELOPMENT OF A DENGUE VACCINE –
LOOKING OUT OF THE BOX
Dengue is globally considered as the most important human disease caused by an arbovirus on an
epidemiological context. Dengue virus (DENV) present four genetically and antigenically distinct serotypes (DENV
1-4) and is estimated to be responsible for 390 million infections per year, worldwide. Despite these numbers,
effective antivirals and vaccines are still elusive. Recently, human trials on a particular vaccine candidate have led
to the possible licensing of this vaccine for human use. However, limitations of this vaccine application in certain
specific populations have raised questions about its real effectiveness. In such scenario, it is prudent to keep on
going research on alternative dengue vaccine strategies. We have developed two different approaches looking for
an effective dengue vaccine. The use of Modified Vaccinia virus Ankara (MVA) as a vaccine vector has showed
promising results in the past and, thus, we constructed and tested four recombinat MVA variants expressing the
DENV-3 E protein and tested their immunogenicity in mice. In such settings, good antibody and cell-mediated
responses were obtained, and protection was observed after mouse challenging with a mouse-adapted DENV
strain. Alternatively, we have also designed a nanoconjugate immunogen based on the association of DENV
recombinant proteins to Gold Nanorods (GNR). Inoculation of this vaccine into mice generated important antibody
and cell mediated responses. Together, these represent alternative dengue vaccines that should be further
developed while a final and cost-effective vaccine solution to dengue is still uncertain.
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Giseli Klassen
Federal University of Paraná,
Brazil
DNA METHYLATION IN CANCER AND IMPLICATIONS FOR THERAPEUTICS
DNA methylation is and enzymatically catalyzed covalent modification of DNA, occurring typically in the context of
cytosine-guanine (CpG) dinucleotides. In general, regions with high CpG content, named CpG islands are
demethylated in normal cells, whereas regions with high or low methylation ratio are involved in genes silencing or
uregulation respectively in cancer. CpG islands are located majorly in gene promoters, but recently many
methylated CG sites were found in intergenic and intragenic regions. The role of DNA methylation in body genes
remains poorly understood. The methylation in DNA is dynamic during the life course. It can be responsive to
environmental exposures such as socio-economic positioning, stress and dietary supplements. DNA methylation is
a reversible process and two cytosine analogues 5-azacytidine (5-aza) and 5-aza-2’-deoxycytidine (5-azaD) are
currently in use like anti-tumour agents. However, these agents might cause activation of a panel of pro-metastatic
genes in addition to tumour suppressor genes, which might lead to increased metastasis. In our laboratory we
have studied 5-azaD in breast cancer cell lines and observed up regulation of MMP9 gene by DNA demethylation.
Furthermore fibronectin that is a matrix extracellular protein cause DNA demethylation in MMP2 gene promoter in
culture. Both MMP2 and MMP9 are genes involved in metastasis process in many cancer types. So we are facing
the challenge of understanding how epigenetic modifiers drugs can interacte in cell function and facilitate the
leading cause of death from cancer that are metastasis.
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Glauber Wagner
Federal University of Santa Catarina,
Brazil
ROLE OF PROTEOMICS IN DIAGNOSIS
Proteomic studies are influencing the development of diagnostic methods in different areas through the discovery
of keydisease biomarkers. Proteomics is an array of complex technologies that incorporates protein separation
methods, mass spectrometry (MS), and bioinformatics on a massive scale. Recent advances in MS techniques,
together with whole genome sequencing and the development of powerful bioinformatics platforms are essential
factors for the success of present proteomics studies. Different proteomic platforms have been used for biomarker
discovery, from gel-based to liquid chromatography (LC)-based separations of proteins and peptides, alongside
the corresponding MS detection. We describe here a combination of top-down and bottom-up proteomic
approaches applied by our group to the study of microorganisms and toxins. We applied gel-based proteomics and
a top-down whole organism for rapid identification of biomarkers using MALDI-TOF MS. We then applied bottomup gel-independent shotgun proteomics approaches based on LC-MS/MS studies as the next step for biomarker
identification. The complexity of the proteomes is due to sequence polymorphisms, posttranslational modifications
and other protein-processing mechanisms. In addition, a proteome’s proteins span a concentration range that
exceeds the dynamic range of any single analytical method or instrument and sample fractionation, which is very
beneficial. Of the many fractionation technologies available, we applied three: affinity purification, LC-based and
GeLC-based separation. The different fractions were digested and the resulting peptides were analyzed by MS
and tandem-MS; label-free quantitative data was obtained. MS/MS data were searched against the entire NCBI
database using the Mascot search algorithm to generate a list of valid proteins. Either common or unique
biomarkers detected were validated. Proteomics will profoundly improve the diagnosis, prognosis, treatment and
prevention of diseases.
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Ian Castro-Gamboa
UNESP,
Brazil
THE FASCINATING WORLD FROM SENNASPECTABILIS RHIZOSPHERE: EXPLORING
MOLECULAR DIVERSITY USING NMR DEREPLICATION TECHNIQUES.
The establishment of new and innovative analytical methods that may let to molecular information towards the
composition of complex natural mixtures is critical on bioprospection programs. Our research group “NuBBE” has
incorporated the use of molecular virtual design using Nuclear Magnetic Resonance (NMR) aiming to increase the
understanding of molecular relationships on dynamic natural matrixes and synergism effects of highly active crude
extracts. Additionally, to speed up the selection of promising biologically active molecules, we have incorporated
the use of chemometricsand multivariate analysis towards dereplication. Recently we have produced a collection
of microorganisms from Sennaspectabilis’srhizosphere, an interesting plant studied along the years that afforded a
series of piperidine alkaloids that, when chemically modified, showed promising acetylcholinesterase activity.
Interestingly, some filamentous fungi isolated from this fascinating niche, produced some metabolites that may be
considered building blocks for those alkaloids and thus, opening new possibilities towards the production of these
promising compounds.
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Jorge Carlos Santos da Costa
Oswaldo Cruz Foundation,
Brazil
HEALTH AND DEVELOPMENT: THE STRATEGIC ROLE OF FIOCRUZ
Health should not be handled outside the context of national development and industrial policy. The Federal
Constitution afirms that “Health is everyone’s right and duty of the State, guaranteed by social and economic
policies aimed at reducing the risk of disease and other health problems and the universal and equal access to
actions and services for its promotion, protection and recovery. “ In this context the Ministry of Health (MH) has
established policies to strengthen national production base, in order to increase access to medicines and supplies
to health, in addition to strengthening the Unified Health System, the so-called SUS.
The Brazilian Productive Development Partnerships (PDP) program was created in 2007 with a mission of
produce, disseminate and share knowledge and technologies to strengthening and consolidation of the SUS and
contribute to the promotion of health and quality life of the population, the national dynamics of innovation
and to reduce social inequalities, with the defense of the right to health as core values. PDPs stimulates the
technology transfer (TT) of strategic products for SUS from private sector to the public laboratories. Since 2007
The Oswaldo Cruz Foundation, federal institution linked to the MH has been actively participating of this on going
public health policy, with drugs and vaccines PDPs of national interest.
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José Fernando Perez
Recepta | Biophama
THE CHALLENGES AND OPPORTUNITIES OF BIOTECHNOLOGY IN BRAZIL: THE
RECEPTA BIO CASE
With activities dedicated to research, development and clinical trials of monoclonal antibodies for use in cancer
treatment, Recepta Biopharma seeks to use technological innovation in biotechnology to meet the needs of society
and the Brazilian market and produce effective medicines and less expensive treatments.
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Joseph Nicolazzo
Monash Institute of Pharmaceutical Sciences,
Australia
BRAIN BARRIER TRANSPORT IN ALZHEIMER’S DISEASE: IS IT INCREASED OR
DECREASED?
The blood-brain barrier (BBB), the endothelial lining of cerebral microvessels, forms one of the major obstacles to
the delivery of therapeutics into the central nervous system (CNS). Due to the existence of inter-endothelial tight
junctions and efflux transporters, many therapeutics fail to reach their site of action within the CNS. However,
during various diseases, the integrity of the BBB has been demonstrated to be perturbed, potentially resulting in
altered CNS disposition of drugs. In Alzheimer’s disease (AD), there are multiple reports of decreased BBB
expression of membrane transporters, altered expression of tight junction proteins and thickening of the
cerebrovascular basement membrane. However, the overall impact of these changes on the BBB transport of
drugs remains to be fully characterized. This presentation will focus on studies we have undertaken in mouse
models of AD, where we assessed the BBB transport of various drugs and endogenous molecules which traverse
the BBB via different mechanisms. We have demonstrated that the BBB transport of various drugs and
endogenous molecules is reduced in mouse models of AD, and this is correlated to thickening of the
cerebrovascular basement membrane and decreased expression of intracellular carrier proteins, which we have
recently found to be important in BBB transport. These results contradict the general dogma that BBB transport of
drugs is increased in AD, suggesting that CNS exposure of drugs may actually be decreased in AD.
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Jürgen Bulitta
University of Florida,
USA
NEXT-GENERATION ANTIBIOTIC COMBINATION DOSING STRATEGIES TO COMBAT
MULTIDRUG-RESISTANT BACTERIAL ‘SUPERBUGS’
Background: There is a serious global health crisis caused by a severe lack of available antibiotic treatment
options and a dearth of new antibiotics under development against multidrug-resistant (MDR) bacteria (i.e.
‘superbugs’). The US President Executive Office, EU, WHO, IDSA, and the CDC recognize antibiotic resistant
bacteria as one of the three most serious threats to global health. As it is evident that the development of new
anti-biotics cannot keep pace with increasing bacterial resistance, it is essential that we find innovative ways of
how to use our available antibiotics more effectively.
Methods: To provide a tangible option for combating these ‘superbugs’, we employed an integrate experimental
and Quantitative and Systems Pharmacology (QSP) approach. Informed by mechanistic receptor binding data and
in vitro infection models, we developed QSP models to predict the most efficacious combination dosing strategies
to combat MDR Pseudomonas aeruginosa, Acinetobacter baumannii, and Escherichia coli via innovation antibiotic
combination therapies.
Results: Mechanistically optimized double β-lactam combination therapy substantially killed resistant P.
aeruginosa without emergence of resistance due to synergistic receptor binding. Optimized combination dosage
regimens of a carbapenem plus and aminoglycoside yielded substantial bacterial killing and prevention of
resistance of carbapenem-resistant P. aeruginosa and A. baumannii. Our QSP models predicted that the
aminoglycoside enhanced the target site penetration of the carbapenem. Both of these combination dosage
regimens were prospectively validated in the hollow fiber in vitro infection model with prevention of resistance over
7 days. Static in vitro infection model studies in E. coli producing extended-spectrum β-lactamases showed that
mechanistically informed combination therapies of oral antibiotics hold excellent promise to combat these resistant
isolates.
Conclusions: The proposed novel QSP approach successfully predicted mechanistically optimized combination
dosage strategies against MDR Gram-negative ‘superbugs’ which provided substantial killing and prevented
resistance. These combination dosage regimens are based on clinically available antibiotics and should be
prospectively validated in animal infection model for ultimate testing in future clinical trials.
24
Kildare Rocha de Miranda
Federal University of Rio de Janeiro,
Brazil
NANOSCALE DOMAINS OF POLYPHOSPHATES POLYMERS IN ION-RICH ORGANELLES
Understanding mechanisms involved inosmoregulation control in protozoan parasites has been a challenge for
many research groups. Over the past years, a number of key players in cell signaling in trypanosomatid parasites
have been identified. Among these, inorganic polyphosphate (PolyP) polymers have proven to play important roles
in cell physiology, both as an energy source, stored in its constituent phosphoanhydride bonds, and as a polyanion
that might activate a number of physiological processes. A number of methodsfor PolyP localization and
quantification are available, including DAPI-staining followed by microscopic visualization and quantification, PNMR analysis, enzymatic assay using recombinant exopolyphosphatases and analytical electron microscopy
(AEM).From the AEM point of view, X-ray microanalysis combined with elemental mapping as well as energy
filtered TEM have been the most employed techniques carried out to explore the two-dimensional composition and
distribution of (poly)ions (including polyphosphate stores) within cells. In this work, we combined different threedimensional electron microscopy techniques with X-ray microanalysis using more sensitive detectors to generate
three-dimensional nanoscale elemental maps of polyphosphate-rich organelles present in the protozoan parasite
Trypanosoma cruzi. We showed a heterogeneous three-dimensional distribution of ions within the shell of
polyphosphate polymersforming segregated nanochemical domains with an auto exclusion pattern for the cations.
This is the first direct evidence for the asymmetric distribution of cations bound to a polyphosphate polymer, raising
questions about polyphosphate assembly mechanisms and its influence on the functional role of polyphosphate in
cell physiology. In addition, these strategies were used here to explore the three-dimensional elemental distribution
are novel for biological materials and may be applied to future studies in a wide variety of biological samples.
Analytical electron tomography of T. cruziion-rich organelle at nanos cale showing the 3D distribution of
phosphorus, magnesium and calcium. The upper two lines show a central slice from the 3D mapping segmented
by Voltex (Amira) of each element and the combination between these elements to visualize the degree of
localization. 3D models from the same elements are also shown.
25
Mário Steindel
Federal University of Santa Catarina,
Brazil
EXPLOITING THE POLYAMINE METABOLISM OF KINETOPLASTID
INSIGHTS TOWARDS RATIONAL DISCOVERY OF NEW DRUG TARGETS.
PARASITES:
Human diseases such as Leishmaniasis, Chagas disease and African sleeping sickness that affects million people
in developing and under developed countries are caused by pathogenic try panosomatids. Due the availability of
these parasites genomes, species-specific metabolic pathwaysare promising targets to be explored towards
development of specific inhibitors. Nowadays, twoma in approaches for seeking newanti-parasiticdrugs are being
used: the whole-cell screening and the target-based discovery.
Polyaminesare essential precursors for try panothione, the central molecule in volved on metabolic detoxification
of reactiveoxygen species (ROS) in trypanosomatids. Therefore, enzymes of the polyamine
biosynthesispathwaysuch as Trypanothionereductase (TR), Trypanothionesynthetase (TryS) and Spermidine
synthase (SpS), as well as enzymesinvolved on the cysteinemetabolismsuch as cysteinesynthase(CS)
andcystathionineβ-synthase (CβS)are promisingdrug targets towards development of novel therapeutic agents
against leish maniasisand try panosomiasis.
Gene expression, activity, functionalrescue and over expression of CSandCβSwere evaluated
invitrousing
both promastigotes and intracellularamastigotes of Leishmaniabraziliensisunder stress condition sinduced
byhydrogen peroxide(H2O2), S-nitroso-N-acetylpenicillamine (SNAP) orantimonialcompounds.
Our results demonstratea stage-specific increaseof protein expression and activity levels of LbrCS and
LbrCβSresultingin high total thiol levelin responseto bothoxidative andnitrosativestress. Over expression of
LbrCSLbrCβSbyintracellularamastigotesresultedina phenotypetwo-fold more resistant toSbVtreatmentin vitro.
Therescue of CSactivityinT. rangeli, a non-pathogenic trypanosome that lackscysteinebiosyn the sisde novo,
resulted inincreased survival rates of epimastigotesexpressingLbrCSunder stress conditions when compared to
wild-typeparasites. Considering that mammals alsolack thede novo cysteine biosynthesispathway and the fact
thatup-regulationof protein expression and increased levels of LbrCSactivity lead to increased resistance to SbV,
the Leishmania spp. make CSa promising candidatefor rational drug designof selective the rapeuticinhibitors of
New World Leishmania parasites.
26
Maria Pilar Vinardell
University of Barcelona,
Spain
COURSE ON ALTERNATIVE METHODS FOR BIOLOGICAL TESTS
In 1959, Russell and Burch published the book “The Principles of Humane Experimental Technique.” In this book,
they defined humane science with the concept of the 3Rs – replacement, reduction and refinement. Replacement
refers to the use of in vitro and other non-whole animal techniques to address a scientific question. Reduction
expresses the concept of using the lowest number of animals without compromising scientific soundness and
focuses on the experimental designs. Refinement refers to using techniques and approaches that eliminate or
decrease pain and distress experienced by the experimental animals used in research or testing.
Russell and Burch also pointed that good science and animal welfare should go hand in hand. This concept is now
more generally accepted and is a driving force for appropriate experimental designs.
Today, the ideas from “The Principles of Humane Experimental Technique” have been incorporated into national
and international regulations. Russell and Burch changed the debate on the use of animals from a philosophical
matter to a scientific issue. By introducing the principles of refinement, reduction and replacement (3Rs) as goals
and guidelines for the work with animal subjects, they put a high value on the integrity of living beings in scientific
practice.
This is a three days course, were we will discuss the principles of the 3Rs and particularly, the application of the
replacement on the evaluation of the activity and toxicity of new products developed for pharmaceutical
applications.
In this course, we will talk about the validation process and the acceptance of alternative methods by regulators
with different examples. We will study different alternatives developed to replace animals in biological tests and we
will see the last advances in this field with the incorporation of new technologies.
27
Nelilma Correria Romeiro
Federal University of Rio de Janeiro,
Brazil
CHEMOGENOMICS APPLIED TO THE PREDICTION OF THE MODE OF ACTION OF
BIOACTIVE SUBSTANCES FROM NATURAL SOURCES
Chemogenomics is an interdisciplinary strategy for drug discovery that involves the investigation of molecular
libraries searching for ligands able to interact with biological targets of interest, among other applications. In the
process of searching for new ligands or new targets, molecular similarity strategies with fingerprints are used,
among others, including approximations based on targets’ structure, such as docking. In this context, we will
discuss recent examples of the application of chemogenomics coupled with other molecular modeling methods in
the design and discovery of bioactive molecules and the current work under development at LICC-Laboratório
Integrado de Computação Científica/UFRJ Macaé.
28
Nina Karpova
Brazil
TARGETING EPIGENETICS
Growing burden of neuropsychiatric disorders induced by early-life stress requires understanding the mechanisms
of stress influence on brain plasticity and functioning. Alterations in environmental conditions during early stages of
development are implicated in the onset of many pathological conditions. Since recently, much attention has been
focused on environmental risk factors that establish long-lasting epigenetic marks in the genome, impair neural
plasticity and lead to the development of neurological and psychiatric diseases. Anxiety and pathological fear are
among the most common neurological disorders, but their developmental origin is yet to be discovered. Clinical
data reveal that anxiety and fear disorders are most effectively treated by psychotherapy (such as extinction
training) in youth but, if not treated or treatment-resistant, have high co-morbidity with the psychiatric diseases in
adulthood. In rodents, fear memories can be completely erased by extinction training during a postnatal critical
period for development of fear memory network (age P16-P18, the period of high neuronal plasticity) but not after
P21(the period of decreasing neuronal plasticity). We aim at discovering new epigenetic biomarkers (DNA
methylation and microRNAs) of anxiety/fear disorders induced by the recognized perinatal stressor - environmental
contaminant methylmercury. We show that perinatal exposure to the low-dose of methylmercuryalters the critical
period of development of fear memory network and leads to the enhanced context and cue-induced fear memories
after extinction of conditioned fear in young mice. A microRNAs screen identified specific microRNAs, brain
expression profiles of which parallel the state of neuronal plasticity in the fear-conditioned mice. The bioinformatic
analysis detected several common gene targets of identified microRNAs. The analysis of identified gene targets
may allow for discovering new molecular pathways implicated in development of anxiety/fear disorders induced by
prenatal stress, such as the exposure to methylmercury.
29
Norberto Peporine Lopes
Brazil
MASS SPCCTROMETRY TECHNIQUES FOR METABOLOMICS AND TISSUES IMAGING
OF NATURAL PRODUCTS
The structural elucidation of increasingly small quantities of organic compounds is a routinely required task of the
chemist, especially in natural product, synthetic organic, biogeochemical, and medicinal chemistry research
laboratories. Among the analytical tools available, there is a wide variety of spectroscopic and spectrometric
techniques. In recent years, developments in these techniques have allowed the analyst to obtain valuable
structural information of a given compound at low concentrations (at high sensitivity) and in an increasingly short
time period.
Each analytical method provides different, complementary structural information about a given compound. When it
comes to mass spectrometry (MS), it is possible to obtain data regarding the molecular weight and/or the
molecular formula of compounds, the presence of heteroatoms, and the presence of functional groups, depending
on the characteristics of the mass spectrometer. Tandem mass spectrometry (MS/MS) is also able to provide
additional structural information through fragmentation of the compound of interest. Therefore, MS and MS/MS are
powerful tools for the structural elucidation of a wide range of organic compounds. In this work we present an the
basic decomposition reaction for natural products from plant and marine organism species and the perspective for
the use in metabolomic studies and the tissues imaging generation for single molecules.
30
Per Artursson
Uppsala University,
Sweden
NEW CELL BASED APPROACHES FOR BETTER PREDICTIONS OF DRUG TRANSPORT
AND CELLULAR DRUG EXPOSURE
Per Artursson, Department of Pharmacy, Uppsala University, 75123 Uppsala, Sweden
Drug transporting proteins influence the ADME profiles and target access of many drugs. It is therefore of great
interest to develop methodologies for quantification of drug transporter expression to understand their influence on
membrane permeability and intracellular drug exposure. In this lecture, approaches for quantitative assessment of
transporter impact will be presented. Methodologies for measurement of transporter abundance and intracellular
free concentrations will be covered and combined with transporter kinetics for more quantitative predictions of
transporter contribution to membrane permeability, intracellular drug exposure and drug-drug interactions. So far,
ADME scientists have mainly used so called targeted proteomics to quantify the expression of individual
transporters after isolation of membrane proteins of variable purity. Technological advancements in mass
spectrometry now allow label free quantification of entire tissue/cellular proteomes without prior isolation of the cell
membranes. Thus, with the label-free approach, not only selected membrane transporters, but also all other
membrane and cytosolic proteins will be covered. By combining protein abundance data with in vitro studies of
transporter kinetics, the individual contribution of multiple transporters can be used to predict e.g. intrinsic
clearance in more complex tissues such as the liver. New cell based methodologies for assessment of intracellular
(free) drug concentrations have recently been developed and will be presented. These methodologies can be
used to study the influence of transport proteins on drug exposure. Recent results will be used to illustrate the
impact of individual uptake and efflux transporters on intracellular drug concentrations and hence access to the
target of interest, whether it is a metabolic enzyme or a receptor. In summary, by taking protein abundance, in vitro
kinetics of individual transporters and intracellular (free) drug concentrations into account, more precise predictions
of transporter impact will result.
31
Peter X. Ma
University of Michigan,
USA
NANOFIBERS AS SCAFFOLDING MATERIALS FOR TISSUE REGENERATION
Regenerative medicine aims to regenerate tissues and organs by utilizing the highly regenerative potentials of
various stem cells, such as embryonic stem cells, multipotent adult stem cells, tissue specific stem cells, and
induced pluripotent stem cells. There is a growing recognition of 3D matrix microenvironments on the fate and
function of stem cells. A key challenge facing regenerative medicine is to rationally design 3D microenvironments
that can recapitulate those in a developmental or healing program to maintain stemness, to accelerate
proliferation, or to direct stem cells to differentiate along a specific therapeutic lineage. Our lab takes a biomimetic
approach to design biomaterial-based 3D microenvironments on the nano- and micro-scales (matrix, signals, and
supporting cells etc.) for stem cells to regenerate tissues. This lecture will focus on nanofiber based scaffolds for
cell and biomolecule delivery to facilitate tissue regeneration. Examples of regenerating dental, craniofacial,
orthopaedic, and cardiofascular tissues will be presented to demenstrate the effectiveness of biomimetic
approaches on the nano- and micro-scales.
32
Pieter Dorrestein
University of California,
USA
DIGITIZING THE CHEMISTRY OF MICROBES AND PEOPLE THROUGH MOLECULAR 3D
CARTOGRAPHY
It is becoming clear that the microbiome plays critical roles in human health but to understand the roles microbes
play and their relationship to the chemistries in our bodies is still poorly understood. UCSD has recently launched
the microbiome center. The Collaborative Mass Spectrometry Innovation Centerplays a key and integrated role in
this center to begin unraveling the chemistry of the microbiome. In this presentation we will highlight the latest
mass spectrometry based tools, including our crowd source molecular annotation platform, to study the chemistry
of microbial interactions and microbiome of the skin, gut and lungs of mice and people in relationship to clinical
information.
33
Rachel Auzély-Velty
CERMAV
France
SMART DELIVERY SYSTEMS BASED ON POLYSACCHARIDE HYDROGELS AND
NANOGELS
The important technological advantages of drug delivery systems and the growing demand for materials with
biocompatibility, biodegradability, and non-toxicity have simulated efforts for engineering new materials based on
new synthetic strategies. In this regard, several approaches have been proposed to synthesize hydrogels to
encapsulate different classes of hydrophilic therapeutic agents such as peptides, proteins and nucleic acids, using
polymers that are biodegradable or that can respond and release their payload upon well-defined stimuli. However,
delivering controlled amounts of small hydrophobic molecules from these hydrophilic matrices remains a
challenge. Water-insoluble organic compounds constitute a great part of the currently available drugs especially in
anti-cancer therapy.
The lecture presents results of our recent studies on the development of smart hydrogels and nanogels (hydrogels
confined to submicrometric dimensions)from biocompatible polysaccharides for hydrophobic drug delivery.
We thus demonstrated that grafting onto hyaluronic acid (HA), a polysaccharide ubiquitous in the body,
thermosensitive ethylene-glycol based copolymers allowed temperature-triggered assembly of HA into nanogels
with diameters < 200 nm.1 These gel particles possess many interesting features for drug delivery, like: facile
formation, tunable size, easy loading of hydrophobic molecules, high selectivity and binding affinity for cancer cells
expressing the CD44 receptor of HA, degradation behavior due to the inherent biodegradability of HA. In addition,
after intraveneous injection in mice, they were shown to enter the blood circulation.2
Moreover, by combining lipid nano particles (LNPs) and carboxymethylcellulose (CMC) hydrogels, we developed
original hybrid biomaterials that are able to provide local delivery of hydrophobic therapeutic agentsin a predictable
and sustained manner.
These new delivery systems offer promising platforms for the controlled release of various drugs under certain
external stimuli (mechanical stress, light irradiation, etc.).
[1]
J. Jing, D. Alaimo, E. De Vlieghere, C. Jérôme, O. De Wever, B. G. De Geest and R. Auzély-Velty, J.
Mater. Chem. B2013, 1, 3883-3887.
[2]
T. F. Stefanello, A. Szarpak-Jankowska, F. Appaix, B. Louage, L. Hamard, B. G. De Geest, B. van der
Sanden, C. V. Nakamura and R. Auzély-Velty, ActaBiomater.2014, 10, 4750-4758.
34
Ricardo Bentes de Azevedo
University of Brasília,
Brazil
NANOMEDICINE – HOW FAR ARE WE?
Nanoscience and nanotechnology are the study and application of things that are in the scale of 1 billionth of a
meter and can be used across in virtually all science areas, such as chemistry, biology, physics, materials science,
engineering, etc…In the last decades nanotechnology has developed in almost all fields of the modern society,
including communication, technology of information, transport, beauty, health care, medicine, among others.
For the field of medicine, nanotechnology has received special attention, since it can be used for drug and gene
delivery, diagnostic and even a combination of treatment and diagnostic at the same time, field called as ther
agnostic. Several nanomedicines are already at the market for the treatment of different diseases, mainly cancer.
However, several other that promises a breakthrough in the way we handle with the people health, are in
development, such as “nanosponges” to absorb toxins and remove them from the bloodstream, use of magnetic
nanoparticles for magnetohyperthermia, light activated drugs, ultrasound activated drugs, nanoparticles to absorb
free radicals, among others.
35
Robert Verpoorte
Leiden University,
Holland
SYNERGY: EASIER TO SAY THAN TO PROVE
In the discussion on medicinal plants often synergy is used as an argument to explain activity, and even to argue
that they are superior to single pure compounds. But how much real evidence is there for synergy? Synergy in
simple words means that 1+1>2. To proof synergy between two compounds for a certain biological activity
isobolograms are used in which one can see that the activity of the mixtures of two compounds is higher than the
sum of the two. Most papers on synergy describe this method to prove synergy of two compounds, however, this
requires that one knows the active compound(s) of a medicinal plant. If these are not known it becomes difficult,
particularly when in bioassay guided fractionation the activity is lost, e.g. when activity is fully dependent on the
presence of two or more compounds. The only solution is a systems biology approach. By measuring the
metabolic profile of different extracts of a medicinal plant or fractions thereof and combining that information with
the results of the bioassays of these samples one may identify the signals that correlate with activity. These
signals may be due to one or more compounds. After identification of these compounds, e.g. after isolation via
metabolomics guided fractionation, one can test these compounds for synergy.
If synergy would play an important role in medicinal plants, the synergistic effect is on the system as a whole, and
thus may have many forms. That means for studies on synergism one should use preferably in-vivo bioassays,
and if possible even apply this approach in clinical trials, as besides synergy also prodrugs may be present in
medicinal plants. Ideal would be clinical trials, which should be possible with already generally used medicinal
plants. Complicating factor could be the principle of personalized medicine, which means that every patient will
have a different treatment. At the same time that is an interesting challenge for a systems biology approach in not
only understanding the mode of action, but also about the symptoms of an ailment and its diagnosis. The omics in
combination with various bioinformatics and statistics tools offer us a great opportunity to enter a completely novel
way to drug development.
36
Thiago Mares Guia
Brazil
NEW TRENDS IN BIOTECHNOLOGY
Global and Brazilian scenarios in pharmaceutical biotechnology – historic overview, market, perspectives
From biosimilars to innovative biologics – is Brazil prepared to catch up and compete globally?
37
Valtencir Zucolotto
Brazil
NANOMEDICINE AND NANOTOXICOLOGY: NOVEL NANOMATERIALS FOR DIAGNOSIS
AND THERAPY
Nanomaterials including nanoparticles, nanowires and nanotubes have been widely applied in biomedical
applications due to their unique/enhanced properties. Manipulation of nanocomposites in conjunction with
biomolecules, for example, is crucial for the development of novel bio-conjugates for applications in
biotechnological areas, especially in Nanomedicine. These nanocomplexes have been explored as novel analytical
tools for diagnosis and therapy, upon conjugation with biomolecules capable of targeting specific organs and cells.
Although the nano-biocomposites may exhibit excellent performance with high affinity and specificity, the
interpretation of the interactions between the biomolecules and the nanoscale sized materials may be
controversial, as well as the potential toxicity exhibited by the latter materials against human health and
environment. This talk focus on the development of novel nanohybrid materials for sensing and immunosensing,
including nanorods and graphene complexes for infectious diseases diagnosis, and for cancer cell imaging and
therapy based upon hyperthermia techniques. Functionalized Au nanoparticles, for example, exhibited excellent
selectivity and affinity toward K562 and HepG2 cancer cells, acting as nanoprobes for optical diagnosis. The
toxicity exhibited by the nanocomplexes against healthy human cells and aquatic organisms will also be
addressed.
38
Vanessa Mosqueira
Federal University of Ouro Preto,
Brazil
NANOTHERAPEUTICS IN THE TREATMENT OF INFECTIOUS DISEASES
Nanomedicine is a large field of interdisciplinary science focused in developing intelligent new medicines in
nanometrical scale of size. This technology is largely applied in cancer research, however it can also be useful in
parasitic diseases chemotherapy. It has been demonstrated that nanocarriers providedless toxic and more
efficacious treatments compared with free drugs in experimental models. The biodistribution of the drugs are
significantly modified by those carriers. The objective of this conference is to show some of the applications of
polymeric biodegradable nanoparticles on the experimental treatment of malaria, Chagas disease, experimental
cancer, infections in cattle, and other diseases by using drugs associated to those nanocarriers. Particularly,
stealth nanosystems, which are able to escape from the massive uptake of the immune system and stay longer in
the blood compartment, without being rapidly degraded or eliminated, will be presented. These advanced
properties of the nanoparticles have been used in our laboratory to produce nanocarriers of antiparasitic drugs and
anti-infectives tested in several experimental models. It involves a plethora of physicochemical techniques and
protocols in biological area to investigate the characteristics of the bioactive nanocarriers and its interactions with
the biological systems in vitro and in vivo. We have also used stealth nanoparticles for the diagnosis of infections
and inflammations using radioactive and fluorescent labeling. One of the recent areas of research is devoted to
study in vivo the toxicological effects of nanoparticles and their risk-benefit relationship. The different polymers and
the chemical nature of the surface of the nanoparticles play an important role on their biological fate. Their effects
on drug pharmacokinetic and biodistribution will be also presented. The advantages and the main challenges of
the nanomedicine in such diseases will be discussed.
39
William R. Folk
University of Missouri,
USA
USING ‘OMICS TECHNOLOGIES FOR VALUING THE SAFETY AND EFFICACY OF
ETHNOMEDICINES – A CASE STUDY
Natural products and related structures will continue to be essential sources of pharmaceuticals and medicines for
much of the world, because of the immense variety of functionally relevant secondary metabolites of microbial and
plant species, and their familiarity and accessibility. The development of powerful 21st century ‘Omics technologies
are greatly expediting characterization of these natural products and derivatives. When combined with robust
ethnomedical studies of traditional medicines, these advances can help ensure critically needed and improved
treatments that are inexpensive, accessible, safe and reliable. However, with growing access to a larger
pharmacopoeia, there is need for special attention to potential antagonisms and risks caused by concurrent use of
natural products used traditionally and newly introduced medicines – and for ensuring good communication occurs
between healthcare providers and the public about them. I will illustrate these points with a case study of
Sutherlandiafrutescens, traditionally used in South Africa for treatment of symptoms of HIV and Mtb infections, and
forchronic diseases such as diabetes and cancer, and about which we have learned much using ‘Omics
technologies. Recent clinical and laboratory experiments suggest that metabolites in Sutherlandiamight interfere
with TB medicines that rely upon oxidants for activity. If confirmed, such interactions would increase development
of Mycobacterial drug resistance, and underscore the need for searching for analogous interactions between other
botanicals and natural products and TB medicines.
40
41
FACTORS ASSOCIATED WITH POTENTIALLY INAPPROPRIATE MEDICATIONS AMONG
HYPERTENSIVE ELDERLY PATIENTS IN THE BRAZILIAN PRYMARY CARE SETTING
Fabiane Raquel Motter
Universidade do Vale do Rio dos Sinos (UNISINOS)
Vera Maria Vieira Paniz
Maria Teresa Anselmo Olinto
Hypertension affects 65% of the brazilian elderly population. Elderly hypertensive patients often require
multiple medications to adequately and appropriately treat their hypertension and associated comorbidities The
use of multiple medications may also lead to the use of potentially inappropriate medications (PIMs), which place
medication users at higher risk for adverse drug reaction. Purpose: To investigate the prevalence and
determinants of PIMs among elderly hypertensive patients. Method: A cross-sectional study was conducted with
elderly (≥ 60 years), who were users of antihypertensive drugs dispensed by the Public Pharmacy of São
Francisco de Paula city in the State of Rio Grande do Sul, Brazil. PIMs were identified in accordance with 2015
Beers Criteria. The association between PIMs and independent variables were analyzed by poisson regression.
The study was approved by Ethics Committee of UNISINOS University (UNISINOS; CEP 10/133). Results: A
total of 336 patients was interviewed, 49,7% (IC95% 44,3 -55,1) of patients had at least one prescribed PIM. The
most commonly PIM was omeprazol (26,0 %) followed by ibuprofen (16,4%) and diclofenac (6,3%). Significant
predictors of PIMs were cardiovascular diseases [adjusted PR 1,63 IC95% 1,11; 2,40], and taking more than five
prescribed medications [adjusted PR 1,63 IC95% 1,18; 2,25]. Conclusion: Our study showed a high prevalence of
PIMs. These results may indicate either physicians' unsatisfactory knowledge of which drug are appropriate to
elderly as well as the limited availability of appropriate drugs in Brazilian primary care. Therefore, the education of
physicians and the establishment of public policies to access to appropriate drugs are necessary to ensure the
elderly population safety and rational pharmacoterapy
Financial Support
CNPQ- Conselho Nacional de Desenvolvimento Científico e Tecnológico
Ethical approval
yes
42
METABOLIMICS AS A TOOL FOR UNDERSTANDING THE EVOLUTION IN
TABEBUIA SENSU LATO
Vanessa Samúdio dos Santos
Universidade Federal de Mato Grosso do Sul (UFMS)
Jean Silva do Vale
Flávio Alves Macedo
In the last decades taxonomists are looking for alternative methods to solve classification controversy.
Advances in chemosystematic and analytical instrumentation, in particular hyphenated systems for
quantitative/qualitative analysis, have speeded these studies and are allowing the application of metabolomic
profiling to the global comprehension of metabolism during the evolution. In this work were used metabolomics
approaches, followed by multivariate analysis methods and phytochemical characterization in Tabebuia s.l.
species that need clarification in classification with the following goals: to compare our results with traditional
taxonomy and molecular phylogeny classification; to evaluate the use of metabolomics approaches as a
chemotaxonomic identification tool of the genera; to interpret the evolution of chemical characters based on the
resulting phylogeny. A high-resolution mass spectrometry was used to monitore the compounds in twenty seven
Tabebuia s.l. specimens, with fifteen Handroanthus and twelve Tabebuia s. str. The results showed that Tabebuia
and Handroanthus have different secondary metabolites storage capacity, the genus Tabebuia has higher levels of
iridoid glycosyl esters linked to a phenylpropanoid as specioside, verminoside and minecoside
while
Handroanthus has accumulated iridoids linked to a simple phenol, neolignans and verbascoside derivatives.
These results reinforce molecular studies that split the Tabebuia genus in a polyphyletic group, the variation in
metabolism must be related to genetic changes during the separation of the genera.
Financial Support
CNPq, CAPES, FUNDECT
43
FLAVONOID-AGLYCONES FRACTIONS FROM Achyrocline satureioides BY HIGH
PERFORMANCE COUNTERCURRENT CHROMATOGRAPHY
Sara Elis Bianchi
Universidade Federal do Rio Grande do Sul (UFRGS)
Eduarda Doneda
Valquiria Linck Bassani
Introduction: Achyrocline satureioides is a native Brazilian plant that presents biologically active
polyphenol constituents, such as quercetin (QCT) luteolin (LUT), 3-O-methylquercetin
(3OMQ)
and
achyrobichalcone (ACB), a new molecule recently isolated by our research group. The therapeutic potential of
these flavonoid-aglycones reveals the interest to obtain an enriched fraction or to isolate them in higher quantity
by High Performance Countercurrent Chromatography (HPCCC) for further biological tests. The aim of this study
is to propose both extraction (Sohxlet) and isolation (HPCCC) methods for obtainment enriched flavonoidaglycones fractions from A. satureioides inflorescences, especially with high ACB content. Methods: Flavonoidaglycones extraction was carried out using Sohxlet apparatus, where inflorescences from A. satureioides were
successively extracted with the following solvents: hexane, ethyl acetate and methanol. Extractive solutions were
concentrated by evaporation under reduced pressure. The isolation from the dry extract was carried out using
HPCCC with solvent systems composed of hexane:ethyl acetate:methanol:water. The fraction containing ACB
was evaporated under reduced pressure and the ACB concentration evaluated. Results and Discussion: The
fractions obtained with ethyl acetate, after prior extraction with hexane (Soxhlet) presented higher flavonoidaglycone concentrations (29.50%, w/w). HPCCC was able to separate the flavonoid-aglycones. A fraction
containing 6.63% (QCT), 2.84% (LUT), and 15.43% (3OMQ) was sucessfuly obtained. ACB was separated in a
single cycle, producing a fraction containing 60% (w/w) of this molecule. Conclusions: The proposed extraction
method followed by HPCCC isolation showed to be able to produce high content flavonoid-aglycone fractions,
especially ACB. These excellent results reveal the potential of the method for produce enriched fraction or
isolated flavonoids for biological tests.
Financial Support
CNPq/Brazil and doctoral scholarship from CAPES/Brazil.
44
ANTI-Trichomonas vaginalis, ANTIBIOFILM AND ANTIMICROBIAL ACTIVITIES OF
45
BIOACTIVE METABOLITES PRODUCED BY MARINE
SPONGE-ASSOCIATED FUNGI
Rodrigo Campos
Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil. Centro de Biotecnologia do Estado de Rio
Grande do Sul, Porto Alegre, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil.
Franciane Rios Senger
Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil and Centro de Biotecnologia do Estado de
Rio Grande do Sul, Porto Alegre, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil.
Melissa Landel
Universidade Federal de Alagoas, Alagoas, Brazil
Camila Braz Menezes
Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil
Graziela Vargas Rigo
Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil
Laura Nunes Silva
Danielle Silva Trentin
Alexandre José Macedo
Tiana Tasca
In recent years, metabolites isolated from marine fungi has attracted considerable attention due to
unique chemical structures with diverse biological activities, encouraging further research in the area.
Trichomoniasis is the most prevalent non-viral sexually transmitted disease worldwide, with an increased number
of metronidazole-resistant clinical isolates. Infections caused by bacteria with different resistance mechanisms
represent a major challenge to the current public health. Biofilm formation is considered an important virulence
factor that contributes with persistence of many infections. Taking in consideration these problematic infections,
the aim of this study was to evaluate the anti-T. vaginalis, antimicrobial and antibiofilm activities of metabolites
produced by fungi associated with marine organisms. After screening of 14 fungi, we found that organic fractions
of the fungus Aspergillus niger and Trichoderma harzianum/Hypocrea lixii complex were active against: (i) T.
vaginalis ATCC 30236 and metronidazole-resistant clinical isolate (TVLACM2R); and (ii) Staphylococcus
epidermidis ATCC 35984. Only the organic fraction of A. niger and A. tubingensis demonstrated activity against
Pseudomonas aeruginosa ATCC 27853. These fractions presented antibiofilm activity related with cell death. The
ability to eradicate S. epidermidis biofilms was seen only for Aspergillus flavus organic fraction. Although no
hemolysis was observed in all fractions, cytotoxicity evaluation against HMVII and Vero cell lines showed that
only the organic fraction of A. niger presented cytotoxic effect. In the evaluation of in vivo toxicity, using the
Galleria mellonella larvae model, none of the tested samples caused a reduction in the insect survival. Preliminary
chemical studies points to the presence of peptides. Overall, the range of activities reported confirms the potential
of marine fungi to produce bioactive molecules.
Financial Support
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq/Brazil) Marine Biotechnology Program (Rede MarAtivo, grant 408578/2013-0) and CAPES
for fellowships.
Ethical approval
UFRGS Ethical Committee, number 18923
46
Pharmacokinetic profile of LASSBio-1736 cysteine protease leishmanicidal in rats
Alcides José Martins Parisotto
Fundação Universidade Federal do Pampa - UNIPAMPA (UNIPAMPA)
Carlos Eduardo da Rosa Silva
Barbra Katiucia Moraes
Sandra Elisa Haas
Marina Alves Amaral
Laboratório de Avaliação e Síntese de Substâncias Bioativas - Universidade Federal do Rio de Janeiro,UFRJ, Brasil
Eliezer de Jesus Barreiro
Lidia Moreira Lima
Teresa Dalla Costa
Introduction: Chemotherapy of the Leishmaniasis disease is based on drugs which have low efficacy
and/or high toxicity, besides the increasing development of resistance. Among the compounds synthesized in
Brazil highlights the compound LASSBio-1736. This study aimed to characterize the pharmacokinetics of the
compound LASSBio-1736 in rats. Methods. The protocol of this animal study was approved by the Animal Use of
Ethics Committee of the Federal University of Pampa (No. 23/2013). . LASSBio-1736 was administered to male
Wistar rats in the doses of 3.2 mg/kg bolus intravenous. Individual plasma-concentration were determinated by
HPLC-UV and the profiles were evaluated by non-compartmental and compartmental analysis (Scientist 2.1,
Micromath®). Results and discussion. Model discrimination was assessed by the individual analysis of plasma
profiles and most of the profiles were best fitted to a three-compartment model resulting in an MSC of 2.7 ± 1.3
and correlation coefficient of 0.9463 ± 0.056. The pharmacokinetic parameters estimated by the compartmental
analysis were not statistically different (α = 0.05) from those determined by the independent analysis model,
confirming the suitability of the three-compartment model to describe the experimental data. LASSBio-1736 had a
long half¬-life (t1/2 = 25 ± 8.62 h), low clearance (CL = 0.05 ± 0.007 Lhkg-1), and low volume of distribution (Vd =
0.9 ± 0.21 Lkg-1). Conclusion: The pharmacokinetic parameters showed by LASSBio-1736 was characterized
high half-life time, low clearance and low volume of distribution comparable to conventionally used drugs.
Financial Support
CNPq/Brazil, INCT-INOFAR, UNIPAMPA, PPGCF and FAPERGS/Brazil
Acknowledgments
Grupo de pesquisa em nanotecnologia e toxicologia e grupo de pesquisa em farmacoologia
Ethical approval
Animal Use of Ethics Committee of the Federal University of Pampa (No. 23/2013).
47
INHIBITORY POTENTIAL EFFECT OF Trichilia catigua A. JUSS ON IL-6 SECRETION
Fernanda Gomes Beserra
Universidade Federal de Pernambuco (UFPE)
Marina Ferraz Cordeiro
Gabriela Souto Vieira de Mello
Renata Longhini
Universidade Estadual de Maringá (UEM)
João Carlos Palazzo de Mello
Ivan da Rocha Pitta
Moacyr Jesus Barreto de Melo Rêgo
Background: Popularly known as Catuaba or Catiguá, the Trichilia catigua A. Juss is an abundant plant
in Paraná, Brazil. This kind of plant generally includes secondary metabolites, such as tannins which presents
some biological activities, for instance antifungal, anti-microbial and anti-inflammatory.Thus, this study aims to
evaluate IL-6 cytokine inhibition after treatment with crude extract (CE), aqueous extract (AqE) and ethyl-acetate
fraction (EAF) of Trichilia catigua. CE, AqE and EAF were obtained from barks of this plant specie. Methods:
Nine animals (BALB/c mice) with 45day-old were used to evaluation of IL-6 inhibition. Cytotoxic assay with
BALB/c isolated splenocytes were evaluated by MTT method. After this analysis, splenocytes were cultured for
48 hours (2x106 cells/ml/well) and exposed to 5µg/ml, 10µg/ml, 50µg/ml and 100µg/ml of CE, AqE and EAF.
Untreated well was used as negative control. Concavaline A (ConA) was used as stimuli. Methylprednisolone
was used as positive control at 100μM. After culturing, the supernatant removed was used for measurement of IL6 cytokine expression by sandwich ELISA. Right after experiment the cytokine were analyzed using
nonparametric Wilcoxon test with p significant values < 0.05. Results and discussion: CE, AqE and EAF were
not toxic to splenocytes until 100µg/mL. CE at 50µg/mL, AqE and EAF at 100 µg/mL significantly reduces IL-6 (p
= 0,0117; p = 0,0078; p = 0,0195, respectively). At 50 µg/mL and 100 µg/mL, the activity of CE, AqE and EAF
were most effective in relation to ConA and demonstrated IL-6 inhibition. These extract and fractions were
previously analyzed and presented tannins in its compositions. Previous studies revealed tannins immunodulatory
activity. These results can be associated with tannins presence in CE, AqE and EAF. Conclusion: CE, AqE and
EAF were able to significantly reduce IL-6 levels.
Financial Support
CNPq, INCT_if, CAPES, FACEPE
Ethical approval
CEUA CB-UFPE: 23076.041556/2015-62
48
EVALUATION OF STABILITY AND ANTIFUNGAL ACTIVITY OF NANOEMULSIONS
CONTAINING Eucalyptus globulus OIL
Aline Ferreira Ourique
Programa de Pós-Graduação em Nanociências, Centro Universitário Franciscano, UNIFRA, Santa Maria, RS, Brazil.
Samantha Nunes de Godói
Curso de Biomedicina, Centro Universitário Franciscano, UNIFRA, Santa Maria, RS, Brazil.
Priscilla Maciel Quatrin
Michele Rorato Sagrillo
Curso de Biomedicina, Centro Universitário Franciscano, UNIFRA, Santa Maria, RS, Brazil.
Roberto Christ Vianna Santos
Introduction: The Eucalyptus globulus essential oil has a great antimicrobial potential against a broad
spectrum of microorganisms that instigate a constant search for new therapeutic modalities. However, this oil is
susceptible to processes such as oxidation and thermal degradation, as well as having volatile and waterinsoluble substances, its making their use in pharmaceutical formulations a challenge. In this context, an
alternative to overcome this problem may be the association this oil in nanoparticles. So the objective of this
study was to evaluate the stability and antifungal activity of nanoemulsions containing Eucalyptus globulus oil.
Methods: For this, a formulation stability evaluation was performed to verify its durability and better weather
conditions of storage. After preparation, the formulations (n=3) were stored at three temperature (under
refrigeration, in a climatic chamber and ambient temperature), and evaluated the following parameters from 0 to 90
days: average particle size, polydispersity index, zeta potential and pH . Later they applied the macrodilution and
death curve techniques for antifungal activity check. Results and discussion:With respect to the stability study
our results showed that the best weather conditions of storage was under refrigeration, maintaining its size and
polydispersity index of approximately 75 nm and 0.2, respectively, zeta potential around -7 mV and acidic pH.
Also it was found that Eucalyptus globulus-loaded nanoemulsions were able to inhibit the growth of Candida
albicans, Candida tropicalis and Candida glabrata in concentrations ranging between 22.5 and 1.4 mg.ml-1.
Conclusions: In conclusion, is was observed that the best storage condition of the formulations was refrigerated
and that this nanoparticles demonstrate antifungal activity, consisting of potential therapy using essential oils.
Financial Support: CNPq, CAPES.
Financial Support
CNPq, CAPES
Acknowledgments
Centro Universitário Franciscano, Programa de Pós-Graduação em Nanociências
49
MICRO/NANOSTRUCTURED SYSTEMS FOR COLON SPECIFIC RELEASE OF
METHOTREXATE: EXPERIMENTAL AND COMPUTATIONAL STUDIES
Aline Martins dos Santos
School of Pharmaceutical Sciences, São Paulo State University, Araraquara, SP, Brazil
Flávia Chiva Carvalho
School of Pharmaceutical Sciences, Federal University of Alfenas, Alfenas, MG,Brazil
Deiver Alessandro Teixeira
Federal Institute of Mato Grosso, Cuiabá, MT, Brazil
Maria Palmira Daflon Gremião
School of Pharmaceutical Sciences, São Paulo State University, Araraquara, SP, Brazil
Methotrexate (MTX) is pH-dependent solubility drug. Development of micro/nanosuspensions is a
promising strategy for formulations with drugs that have limited solubility, due to benefits as size reducing of drug
which may lead to the increase of the bioavailability. The aim of this work was to characterize
micro/nanosuspensions of MTX, as well as evaluate the influence of different acid and base proportions and
interaction energy on particle size. Micro/nanoparticles were obtained by Bottom-up method using acid-base
reactions and were characterized by size, polydispersity index and zeta potential. Dissolution profiles were
obtained using USP apparatus II (paddle) at 50 rpm, temperature of 37 ± 0.4 ºC and 900 mL of medium (0.1 M
phosphate buffer pH 6.8). Interaction energy of the particles was evaluated by means of computational
calculations. Experimental results showed that MTX nanoparticles were formed using the lowest HCl proportion
during the drug precipitation, exhibiting a mean particle size between 131.60-186.53 nm with a good homogeneity.
However, the higher HCl proportion during the drug precipitation, the greater the particle size values resulting on
micrometric particles. Zeta potential suggests depending on the micro/nanosuspension pH, the surface charge of
particles can vary from positive to negative. In vitro release profiles showed that micro/nanosuspensions
contributed to the increase of the drug release rates when compared with raw material. Computational results
showed that the MTX clusters in zwitterionic and cationic states (acidic pH) presented higher energy of interaction
between particles, favoring aggregation. The set of results indicate that the different conditions of precipitation
was possible to modulate the particle size to the micro/nano range. Although molecular simulation showed a
microaggregation trend, it was found in this study experimental conditions for obtaining nanosuspension, which is
important to target the colon release.
Financial Support
FAPESP and CAPES.
Acknowledgments
FAPESP and CAPES.
Ethical approval
FAPESP and CAPES.
50
SOLID DISPERSION WITH HECOGENIN ACETATE AND HPMC: AN ALTERNATIVE TO
IMPROVE SOLUBILITY AND ANTIHYPERALGESIC EFFECT IN NEUROPATHIC PAIN
MICE MODEL
Carlos Demócedes Luís de França Almeida Moreira
Universidade Federal do Rio Grande do Norte (UFRN)
Walter Ferreira da Silva Júnior
Jonas Gabriel de Oliveira Pinheiro
Euzébio Guimarães Barbosa
Lucindo José Quintans Júnior
Universidade Federal de Sergipe (UFS)
Jullyana de Souza Siqueira Quintans
Adriano Antunes de Souza Araujo
Ádley Antonini Neves de Lima
Introduction: The literature already reports Hecogenin Acetate (HA) analgesic profile that can be
enhanced with the formation of Solid Dispersion (SD) with hydrophilic polymers. The present study aims the
development of SD with HA and Hydroxipropylmethylcellulose (HPMC) evaluating physicochemical and in vivo
parameters. Methods: The samples were obtained by two methods: physical mixture (PM) and Kneading (KND) in
1:1 (w/w) proportion. After this, physicochemical characterization was made by Infrared Spectroscopy (FTIRATR) with Spectral Correlation by ad hock algorithm, Scanning Electronic Microscopy (SEM) and Powder X-Ray
Diffraction (PXRD) and after evaluating in vivo study. This study was made with Swiss Mice inducing Nerve
Crush Injury (NCI) and treating it pharmacologically to evaluate the measurement of thermal and mechanical
hyperalgesia in mice and also assess the muscular strength on grip strength apparatus. Results and Discussion:
The PXRD showed the crystalline profile of HA while HPMC presents low intensity crystalline reflections,
showing an amorphous behavior that is evident in SD obtained by the KND method. Proving that this method is
efficient to reduce crystalline profile and finally enhance HA solubility. The SEM images refutes the simple
monoclinic aspect of HA crystals and the amorphous morphology of HPMC. SD obtained by KND notorious
changed HA crystals proving an interaction between HA and HPMC. FTIR analysis evidences a strength
interaction between HA and HPMC with significant changes in HA spectra, proved by the spectral correlation with
that overlaps the HA and SD's spectra and shows the low similarity. The measurement of mechanical and
thermal antihyperalgesia was notorious improved when HA was incorporated with HPMC. This effect of HA
seems to act through opioid receptors which SD improved these pharmacological activity. Conclusion: Thus, SD
is an interesting alternative to improve both solubility and antihyperalgesic profile of HA.
Financial Support
CNPq, FAPITEC
Acknowledgments
Pedro Hentique Antunes de Azevedo (in memorian)
Ethical approval
Approved by the Animal Care and Use Committee at UFS/Brazil (40/14)
51
DEVELOPMENT OF SEMISOLID FORMULATION CONTAINING NANOENCAPSULATED
SUNSCREENS AND COATED-MICRONIZED TITANIUM DIOXIDE AS DRYING EXCIPIENT
Karina Paese
Pharmaceutical Nanotechnology Graduate Program, University of Rio Grande do Sul, Porto Alegre, RS, Brazil
Carolina Araujo Cirne
Department of Production and Control of Medicines, University of Rio Grande do Sul, Porto Alegre, RS, Brazil
Denise Soledade Jornada
Pharmaceutical Sciences Graduate Program, University of Rio Grande do Sul, Porto Alegre, RS, Brazil
Julia Scherer Santos
Pharmaceutical Nanotechnology Graduate Program, University of Rio Grande do Sul, Porto Alegre, RS, Brazil
Adriana Raffin Pohlmann
Silvia Stanisçuaski Guterres
Introduction: The application of polymeric nanostructures in photoprotective formulations is of great
interest in cosmetic industries because of its several potential benefits, such as improving SPF and photostability,
controlling release profile, increase sunscreens performance for promoting the formation of a film on the surface of
the skin and reducing side effects. The aim of this work was to obtain a spray-dried nanostructured system
containing simultaneously the sunscreens octyl methoxycinnamate (OMC), octyl triazone (OCT), bemotrizinol
(BMT) and titanium dioxide (TiO2). Methods: The nanocapsules, prepared by interfacial deposition of polymer,
were characterized prior to spray-drying process. The suspension was dried in a spray-dryer with TiO2 as drying
excipient. The diameter was determined by laser diffraction, the sunscreens content was measured by HPLC and
the presence of nanocapsules in the produced powder was observed by scanning electron microscopy. The
powder was incorporated in emulsions and the rheologic behavior, sunscreens content, pH and photoprotective
capacity were determined. Results and discussion: The powder was successfully prepared and characterized
before incorporation in the emulsion formulation. Emulsion containing spray-dried nanoencapsulated sunscreens
presented a pH value of 6.37 ± 0.08; a sunscreen content (mg mL-1) of 0.99 ± 0.13 for OMC, 2.38 ± 0.34 for OCT
and 1.37 ± 0.063 for BMT; and pseudoplastic rheological behavior. The UV blocking activity study under UV
radiation showed that the nanocoating of TiO2 microparticles increased significantly (p<0.05) the photoprotective
activity of the formulation. Conclusion: The novelty of our strategy was showed by its combining advantages of
producing nanocoated powders instead of only nanocapsule suspensions, and presenting the potentiality of TiO2
as a feasible spray-drying excipient for this application, which may also act as photoprotective agent.
Financial Support
CNPq
Acknowledgments
CNPq, CAPES, FAPERGS
52
Polymer concentration effect in poly(caprolactone) nanoparticles surface chemistry
evaluated by Time-Domain Nuclear Magnetic Resonance
Marina Rodrigues Tavares
Instituto de Macromoléculas Professora Eloisa Mano, Universidade Federal do Rio de Janeiro - IMA, UFRJ
Lívia Rodrigues Menezes
Diego de Holanda Saboya Souza
Maria Inês Bruno Tavares
The development of controlled drug delivery systems (DDS) using polymers has been growing on the
last years. In addition, polymeric nanoparticles became an important topic discussed in pharmaceutical issues
aiming to control drug release, to target these systems to a specific tissue besides reducing therapy adverse
effects. Moreover, particles surface chemistry has been studied due to its influence in avoiding macrophage
recognition in addition to prolong nanoparticles circulation time, which increases the chance of nanosystems to
reach the target tissue. In this context, surface modifications became necessary. In this study, nanosystems with
different poly(caprolactone) (PCL) concentrations were prepared by nanoprecipitation method and its coating with
pluronic, a triblock amphiphilic copolymer made of hydrophilic ethyleneoxide (EO) and hydrophobic propyleneoxide
(PO) units, was evaluated aiming to observe alterations in systems mobility. All systems were evaluated by XRay Diffraction (XRD), Time-Domain Nuclear Magnetic Resonance (TD-NMR) and Dynamic Light Scattering
(DLS). We could observe that as PCL concentration increases, particle size and zeta potential modulus also
increases. Furthermore, dipolar interactions between PCL and pluronic have influence in systems mobility, which
increases with PCL concentration. Larger particles may lead to slower release rates and, furthermore, increasing
PCL rigid fraction may also delay drug release, if hydrophobic drugs are used. Finally, the increase in zeta
potential modulus indicates a possibility to obtain a greater stability since it reduces the chance of nanoparticles
aggregation.
Financial Support
CNPq and CAPES
Acknowledgments
CNPq and CAPES
53
SEMI-SYNTHESIS OF IRIDOIDS PROSPECTED IN PLANTS NATIVE TO RIO GRANDE
DO SUL
Maria Helena Vendruscolo
Maria Luisa Nunes Diehl
Glória Narjara Santos da Silva
Simone Cristina Baggio Gnoato
Gilsane Lino von Poser
Introduction: Iridoids are secondary metabolites restricted to dicotyledonous angiosperms. These
compounds are divided into carbocyclic iridoids and secoiridoids and usually occur in the glycosylated form. From
various plants native to Brazil several iridoids were isolated and biological activities, such as neuroprotective,
cardiovascular, antitumor and immunomodulatory have been reported. In this study, we focus on developing
potential neuroprotective compounds through pharmacomodulations in iridoid skeleton. Methods: The species
Escallonia bifida Link & Otto (Escalloniaceae) was subjected to maceration exhaustively with ethanol. The
aqueous fraction of ethanolic extract was submitted to fractionation by column chromatography over silica gel 60
for the isolation of a main iridoid (1). This compound was submitted to acylation using acetic anhydride/DMAP in
dichloromethane, leading to the formation of 1a. The acylated compounds were subjected to bromination reaction
at C-7 position of the cyclopentane ring using NBS to afford the compound 1b. Results and Discussion: The main
compound isolated from this plant was identified as asperuloside (1). The compound obtained by semi-synthesis
(1a) did not show the characteristic 1H NMR spectrum (D2O, 60 MHz) signals of βDglucose (3.92 - 3.20 and
4.67) indicating that the sugar unit was successfully acetylated. In the 1H NMR spectrum of the compound 1b the
non-appearance of the signal at D 5.75 ppm indicates the bromine substitution at the C-7 position. Conclusions:
Asperuloside had successfully been modified at the C-7 position and the glycosyl moiety, resulting in the
derivatives 1a and 1b. The pharmacomodulation performed aims to obtain more lipophilic compounds in order to
development new compounds derivated from natural sources for neurodegenerative diseases.
Financial Support
The authors are grateful to the funding agencies FAPERGS, CAPES, CNPq for the financial support.
54
Chemogenomics Studies Aiming at the Identification of Natural Products as Potential
Ligands of Matrix Metalloproteinase 9 (MMP-9)
Priscila Baltazar Gonçalves
Universidade Federal do Rio de Janeiro (UFRJ)
Vitor Won-Held Rabelo
Stella Schuenck Antunes
Nelilma Correia Romeiro
Title: Chemogenomics Studies Aiming at the Identification of Natural Products as Potential Ligands of
Matrix Metalloproteinase 9 (MMP-9). Authors: Gonçalves, PB; Rabelo VW; Antunes, SS; Romeiro, NC.
Affiliation: PPG-ProdBio-Programa de Pós-Graduação em Produtos Bioativos e Biociências, Universidade Federal
do Rio de janeiro, Campus Macaé Professor Aloísio Teixeira, RJ, Brasil. Introduction: Chemogenomics has been
used to predict the mechanism of action (MOA) of molecules for which only phenotypes are known. In this work,
we searched for Natural Products (NPs) from the Brazilian Biodiversity which could hit targets related to Chronic
Obstructive Pulmonary disease (COPD), such as Matrix Metalloproteinase 9 (MMP-9). The search for new
bioactive NPs for COPD is justified by the need of new drugs to treat this multifactorial disease and by the
reported activity of NPs against COPD. Methods: The compounds were obtained from NUBBE and standardized
using Standardizer v. 6.2.1 for chemogenomics analysis. MOAs were predicted using PIDGIN and Docking
Studies were performed with AutoDock Vina v. 1.5.6. The 3D structure of MMP-9 was obtained from PDB, ID
4XCT. Visualization of the docking results was done with Pymol v. 0.99 and Discovery Studio 3.1. Results and
Discussion: Chemogenomics analysis suggested 100 potential MMP-9 ligands. Initially, 20 molecules were
selected for Docking, including Caffeic, Salicylic and Protocatechuic acids, which have already been reported as
MMP-9 inhibitors. However, Arboreumine and Dibenzyl sulphide, for example, have not. Docking studies showed
similar interactions in the active site for the analyzed NPs. Conclusions: Our studies showed some results that
are supported in the literature, validating the methodology and pointed out new NPs as potential MMP-9 ligands,
for which the experimental assays are not available yet, such as Arboreumine and Dibenzyl sulphide. These NPs
can have their MOAs further confirmed in enzyme inhibition tests.
55
TOXICITY ASSAY OF ETHANOLIC EXTRACT OF Xanthium cavanillesii THROUGH
Artemia salina TEST
Thainara de Andrade Fortes
Faculty of Pharmacy, Integrated Regional University of High Uruguay and Missions, Santiago Campus
Augusto Cezar Neves Mazzui
Thayse de Freitas Oliveira
Lenise de Lima Silva
Health Sciences Departament, Faculty of Pharmacy, Integrated Regional University of High Uruguay and Missions, Santiago Campus
Amanda Leitão Gindri
Pharmaceutical Sciences Graduate Program, Federal University of Santa Maria; Health Sciences Departament, Faculty of Pharmacy,
Integrated Regional University of High Uruguay and Missions, Santiago Campus
The leaves of the species Xanthium cavanillesii (Asteraceae), popularly known as 'carrapicho', are used
to treat ulcers. This plant has high toxicity in their fruits that if ingested by cattle cause death associated with
acute liver failure. The aim of this study was to analyze the potential toxicity of the crude extract of the leaves of
X. cavanillesii on Artemia salina. The leaves of the plant were collected in the city of Itacurubi (RS, Brazil), in
October 2014. The leaves were dried, crushed and extracted in Soxhlet apparatus with ethanol 95% until
exhaustion. The ethanolic extract was dried in rotaevaporator (<40°C). The toxicity test was performed with larvae
of A. salina hatched under the temperature of 30°C in artificial sea water (23 g/l sea salt and 0.7 g/l of sodium
bicarbonate in distilled water). After 24 hours, ten A. salina nauplii were transferred to tubes containing 9 ml of
saline and 1 ml of the sample to be tested. The test was performed in triplicate with two replications. Samples
were tested at different concentrations (from 2000 µg/ml to 10 µg/ml). As a negative control was used 0.5 ml of
ethanol diluted in saline 9.5 ml (simulating the extract dilution), and artificial saline only. As a positive control we
used sodium lauryl sulfate (100, 10 and 1 μg/ml). The yield obtained from the crude extract was 20.21 ± 2.48 %.
Through toxicity test was obtained a value of LC50 (lethal concentration for 50% of A. salina) of 1368.94 µg/ml
(confidence interval: 1289.12 ± 1448.76 µg/ml ). Therefore, this specie has a low toxicity to A. salina, which
motivates the realization of different toxicity studies in order to confirm the safety of its leaves. Additionally, this
result indicates that the leaf chemical constitution may be different from the fruits that have lethal effect in the
cattle.
Financial Support
URI, Campus de Santiago
Acknowledgments
Integrated Regional Foundation (Furi)
56
Echinodorus grandiflorus (CHAPÉU-DE-COURO): A PROMISSING SOURCE OF ANTIARTHRITIS AGENTS
Fernão Castro Braga
Universidade Federal de Minas Gerais (UFMG)
Eliana de Faria Garcia
Mariana Assíria de Oliveira
Luana Pereira Antunes Dourado
Celso Martins Queiroz-Junior
Flávio Almeida Amaral
Danielle Glória de Souza
Mauro Martins Teixeira
Introduction: the leaves of Echinodorus grandiflorus are traditionally used in Brazil to treat several
inflammatory conditions, including arthritis. This study aimed to investigate the anti-arthritis activity of the species
and the association between bioactivity and phytochemical composition. Methods: the effect of extracts and
fractions from E. grandiflorus leaves on TNFα release by LPS-stimulated THP-1 cells was evaluated. A HPLCDAD method was developed and validated to quantify seven compounds in the assayed samples. The antiarthritis activity was investigated in an antigen-induced arthritis model in mice. Results and Discussion: all
assayed samples inhibited TNFα production; extracts prepared from 50% ethanol, water and dichloromethane
elicited the most potent responses, along with a standardized flavonoid-rich fraction (FLAV). trans-Aconitic acid
and isoorientin were the major compounds in some preparations. Polynomial regression analysis showed the
association between the contents of swertiajaponin, swertisin, trans-aconitic and chicoric acids with the anti-TNFα activity of the extracts and fractions. None of the compounds tested alone abolished TNFα release completely,
however some extracts and fractions reached this result, suggesting a synergistic effect between the
constituents. The oral administration of FLAV (0.7-7.2 mg/kg) to previously immunized mice significantly reduced
neutrophil recruitment to the joint cavity and in periarticular tissue. The levels of CXCL1, TNFα and IL-1beta
quantified by ELISA in the periarticular tissue were also diminished in mice treated with FLAV, as well as
mechanical hypernociception. Histological analysis confirmed that FLAV suppressed joint inflammation and
inhibited cartilage and bone destruction. Conclusion: E. grandiflorus has significant in vitro antiTNFα activity and
its flavonoids elicited potent anti-inflammatory activity in experimental arthritis model.
Financial Support
FAPEMIG, CNPq and the European Community's Seventh Framework Programme [FP7-2007-2013]
Ethical approval
Ethics committee certificate number 165/2009
57
METFORMIN POPULATION PHARMACOKINETIC MODEL IN DIABETIC RATS INDUCED
BY STREPTOZOTOCIN
Andressa Braga
Pharmaceutical Sciences Graduate Program of UFRGS
Jaqueline Scheneider Izolan
Daiane Maria Fonseca de Lima
College of Pharmacy- UFRGS
Teresa Dalla Costa
Bibiana Verlindo de Araujo
Introduction: Metformin is the most prescribed drug to diabetic type 2 patients. However, some
pharmacokinetic features in diabetic patients remain not clarified. The population pharmacokinetic (Pop-PK)
modeling is a tool to study the subject variability as well as understand the disease influence on drug's
pharmacokinetics. This work aims to develop a Pop-PK model to evaluate the diabetes mellitus influence on
metformin pharmacokinetic. Methods: Diabetes was induced in male Wistar rats according previous study, which
assumed plasma glucose levels > 250 mg/dL. Healthy (n=5) and diabetic (n=5) rats were anesthetized with ethyl
carbamate (1.25 g/kg, i.p.) and their carotid artery was cannulated, allowing 9 points of blood collection between
0.08 up to 12 hours after i.v. administration of metformin (50 mg/kg). The samples were analyzed using a
validated LC-MS/MS method. Non linear mix effects Pop-PK model was performed using Monolix. Results and
Discussion: A structural Pop-PK model of two compartments with first order elimination process gave the best
goodness of fit diagnostic graphs as well as parameters precision estimate. Pop-PK parameters estimated was
CL 1.17 L/h, V1 1.29 L, V2 1.44 L and Q 0.24 L/h. The inter-individual variability was 23, 31, 30 and 45%
respectively. The covariate glycaemia could explain the inter-compartmental variability (beta=0.007), Wald test
p=0.003. Metformin is dependent of membrane transporters (Organic Cation Transporter - OCT) to be uptaken and
secreted. However, diabetes may decrease OCT expression which could explain the influence of glycaemia on
metformin inter-compartmental clearance in this model. Conclusion: The results of this experiment conducted so
far demonstrated the influence of glycaemia on metformin pharmacokinetic in diabetic rats. On the next steps of
this study the metformin tissue distribution will be evaluated o complement the model described before, in order to
optimize the drug dosing.
Financial Support
CNPq/Brazil and FAPERGS
Acknowledgments
CNPq/Brazil and FAPERGS
Ethical approval
Committee of Ethics in Animal Use - UFRGS UFRGS # 25780
58
DISSEMINATIVE INFECTION MODEL BY Cryptococcus neoformans IN MALE WISTAR
RATS
Keli Jaqueline Staudt
Universidade Regional Integrada do Alto Uruguai e das Missões (URI)
Gabriela Anjos Colombo
Universidade Regional Integrada do Alto Uruguai e das Missões (URI)
Izabel Almeida Alves
Introduction: C. neoformans is an opportunistic pathogen that affects the respiratory tract and CNS,
being able to reach other tissues causing disseminative infection, becoming the major cause of hepatic injury in
HIV patients. Animal models of infections can be useful to better understating the mechanisms involved on tissue
changes and its correlation with failures in the treatment with antifungals. Aims: To develop an in vivo
disseminative infection model caused by C. neoformans, designed to pharmacokinetics studies of drugs.
Methods: Wistar rats 45 days old were used. The infection was induced by injection of 100 µL of inoculum (1-105
CFU) intravenously in the animal tail's lateral vein (n=6). The infection was established after 10 days post
innoculation. The presence of C. neoformans was investigated in two tissues (the Brain and the Liver) where
histological and microbiological investigations were done. The presence of fungal in tissues was evaluated by
hematoxylin/eosin staining method and the viable cells were determined by counting of colony-forming unit. Liver
function and brain vascular permeability were evaluated, in healthy and infected groups. Liver function was
evaluated (ALT, AST, urea and albumin). Results and Discussion: The CFU courting was homogeneous in both
tissues, with mean inoculation equal to 6.4 ± 0.4 log CFU/gbrain and 4.1 ± 0.5 log CFU/gliver. During histological
analyses have been observe yeasts with characteristic of Cryptococcus. The biochemical data it has been
possible to observe liver damage, because of differences of ALT, albumin and urea between healthy and infected
(p <0.01). The Evans Blue allowed to characterize the improvement on vascular permeability in the brain, possible
sue to the inflammatory response related to the experimental infection. Conclusions: The model was validated
with characteristics similar to observed in the clinical scenario, causing changes in plasma proteins, urea and
vascular permeability.
Financial Support
CNPq.
Ethical approval
Ethics Committee in the Use of Animals UFRGS # 26605; Project name ¿Modelagem Farmacocinética-Farmacodinâmica e avaliação da penetração cerebral de
antifúngicos azólicos em animais sadios e com meningite induzida por Cryptococcus neoformans¿.
59
Identification, characterization and in silico ADMET prediction of Roflumilast
degradation products
mariana santos pinheiro
Gil M. Viana
Caroline da S. Pereiraa
Bárbara de A. Abrahim Vieira
Rita de Cássia E. E. Marins
Lucio M. Cabral
Valeria P. de Sousa
The drug stability in medicines is a matter of concern since it affects the safety and efficacy of
pharmaceuticals. ICH guidelines about stability testing and impurities in new drug substances and final products
describes forced degradation as strategy to determine the stability of the molecule. This approach is even more
relevant when it comes to the long term use of drug therapy as Roflumilast (RFL). RFL belongs to the second
generation of selective inhibitors of phosphodiesterase-4 developed for the treatment of chronic obstructive
pulmonary disease and it does not have monographs in official compendia. In this way, the purpose of this study
was to identify, characterize and in silico ADMET predict RFL degradation products (DPs). RFL was subjected to
stress conditions of photodegradation, alkaline and acidic hydrolysis, oxidative and metallic degradation. After
stress, the DP resulted from the first condition was analyzed by UFLC-QTOF methodology. The DPs resulted
from the other three conditions were subjected to the liquid-liquid extraction with ethyl acetate. All those fractions
were analyzed by TLC and HPLC-DAD-CAD. The products isolated by preparative TLC were characterized by
NMR spectroscopy, HRMS and UFLC-QTOF. Besides that, three kinds of RFL tablets were prepared with the
addition of solid stressing substances: tartaric acid, urea hydrogen peroxide and sodium carbonate. All tablets
were stored into accelerated stability chamber during sixteen months. DPs resulting from the tablets were
analyzed by HPLC-DAD-CAD and UFLC-QTOF. The forced degradation study allowed identify and characterize
eleven DPs. Finally, in silico pharmacokinetics and toxicological parameters of all eleven DPs were performed
using ADMET Predictor software. The identification and characterization in vitro and in silico of RFL DPs enabled
us to understand not only the drug's behavior in stressful conditions, but also its DPs formed.
Acknowledgments
The authors thank the Program for Technological Development in Tools for Health-PDTIS FIOCRUZ for use of its facilities.
60
BIPHASIC SOLVENT SYSTEM SELECT FOR CHIRAL SEPARATION OF PRAZIQUANTEL
BY HIGH PERFORMANCE COUNTERCURRENT CHROMATOGRAPHIC TECHNIQUE
Elisa de Saldanha Simon
Sabrina Laíz Büttenbender
Nadia Maria Volpato
Introduction: High Performance Countercurrent Chromatography (HPCCC) is an interesting technique to
analytical and preparative enantioseparation, by mean of a chiral selector added to the stationary phase.
Praziquantel (PZQ) is racemic anthelmintic drug, however the S-(+)-enantiomer is inactive and associated with
side effects and bitter taste. The aim of this work was to determine the biphasic solvent system (BSS) for
application in enantiomeric separation of PZQ by HPCCC. Methods: To determinate the partition coefficient (k) of
PZQ, diverse BSS were tested changing the volume ratio of the solvents in the following: hexano/ethyl
acetate/MeOH/water and EtOH instead MeOH. An amount of PZQ was dissolved in test tubes containing the preequilibrated BSS. Tubes were shaken and allowed to settle. A volumetric aliquot of the lower phase was diluted
with acetonitrile:water. The amount of PZQ present (Wl) was determinated by HPLC and the drug present in the
upper phase (Wu) was estimated by difference. The k value was calculated as Wu/Wl. Solubility of chiral selector
hydroxypropylβcyclodextrin (HPβCD) was determined in both pre-equilibrated phases. Results and Discussion:
BSS hexane/ethyl acetate/MeOH/water ratios tested 1:1:0.5:1.5, 0.8:1.2:0.8:1.2, 1:1:0,8:1.2, 1:1:1:1, and 9:1:9:1
resulted in k values of: : 0.01, 0.16, 0.20, 0.21 and 5.33, respectively. It was also tested system composed of
EtOH. A BSS in which the drug k value is close to one is considered promisor for future separation by HPCCC.
Solubility tests for chiral selector HPβCD demonstrated that it was soluble in lower phase (stationary) and
insoluble in upper phase (mobile). Conclusions: PZQ k value and solubility of HPβCD chiral selector in the
different BSS tested demonstrated that the composition hexane/ethyl acetate/EtOH/water (1:1:1.1:0.9) is more
suitable for the separation of PZQ through HPCCC technique, using HPβCD as chiral selector.
Acknowledgments
Authors wish to thank Pharmaceutical Sciences Graduate Program, Faculty of Pharmacy, Federal University of Rio Grande do Sul, CNPq and the Ashland for
the provision of HPβCD.
61
Cellular Carriers for Medical Diagnostic Applications
Fernanda Zamboni
Department of Civil Engineering and Materials Science, University of Limerick, Limerick, Ireland
Maurice Collins
Stokes Institute, University of Limerick, Limerick, Ireland
Marie Keays
Stokes Institute, University of Limerick, Limerick, Ireland
Introduction: Microfluidics is an emerging technology used for chemical synthesis and drug screening. It
displays advantages regarding time-efficiency, sampling and waste reduction. Recently, this technology is gaining
prominence as medical diagnostic tools. The aim of this project was to produce hybrid cell carriers from synthetic
and natural polymers designed to create a suitable microenvironment for cellular adhesion and proliferation within
microfluidics droplets, thereby enabling the potential of higher throughput and faster disease diagnosis. Methods:
Porous hybrid cell carriers of PLGA 50:50 ester terminated 26 wt% and hyaluronic acid (HA) 1% were produced
by freeze drying and characterized regarding their physical, chemical, thermal and mechanical properties. Biocompatibility was assessed using L929 fibroblast cells and Alamar Blue proliferation assay at 4h, 24h and 48h of
static culture. Cell laden scaffolds containing 1×106 cells/mL in suspension were injected into the microfluidics
device. Droplets (600nL) were then incubated at 37oC in 5% CO2 and monitored after 24 hours. Results: Within
the microfluidics droplet, scaffolds tend to attract and attach cells to their surface after 24h of incubation by which
two cell morphologies were observed, a round shaped cell and a flat spread shaped cell. Once cells are attached
they pass through the quiescent state of the cell cycle and start proliferating. It is further envisaged that HA,
which is a component of the extracellular matrix, enhance the attachment of fibroblasts to the surface of the
scaffold, by binding to specific cell membrane receptors, such as RHAMM and CD44. Conclusions: HA improves
the biological performance of PLGA cell carriers, demonstrating the possibility of culturing cells within a
microfluidic device. These findings are expected to contribute to the production of high throughput diagnostic
device.
Financial Support
Government of Ireland International Scholarship
62
DIHYDROMYRICETIN-LOADED NANOCAPSULES: ACTIVITY ANTI BIOFILM ON
URINARY CATHETER INFECTED BY Pseudomonas aeruginosa
Ana Julia Dalcin
Centro Universitário Franciscano
Renata Platcheck Raffin
Roberto Christ Vianna Santos
Patrícia Gomes
INTRODUCTION: Nosocomial infections associated with biofilm formation in urinary catheters are
among the leading causes of complications due to high antimicrobial resistance. An alternative is
Dihydromyricetin (DMY), a flavonoid compound extracted mainly from Ampelopsis grossedentata plant, which has
different pharmacological properties. Among them stands out a strong antimicrobial activity of various
microorganisms. However DMY, it has high lipid solubility and consequently low bioavailability. The
nanoencapsulation can contribute to the improvement of characteristics of drugs, for increasing the apparent
solubility and sustained release among other advantages have been reported. The aim of this study was to
evaluate the antibiofilm activity of DMY-loaded nanocapsules in infected urinary catheters with Pseudomonas
aeruginosa. METHODS: Nanocapsule suspensions were prepared by the interfacial deposition of a preformed
polymer. The various formulations were added to the medium containing the infected urinary catheters with
Pseudomonas aeruginosa and incubated for 72 and 96 hours. The antibiofilm activity was evaluated by reading
the absorbance microplate reader at a wavelength of 570 nm. RESULTS AND DISCUSSION: The antibiofilm
activity of the formulations resulted in the eradication of biofilms in both formulations (nanocapsules without DMY
and with drug) during the time of this study. However, within 96 hours the results of the inhibition of biofilm by
DHM nanocapsules were more effective compared with free DHM eradicating 67% of biofilm population in
hospital devices. CONCLUSION: Thus, the nanocapsules of DMY may be used in the eradication of biofilms on
urinary catheters, it may be used as an innovative strategy.
Financial Support
CAPES
Acknowledgments
63
PRELIMINARY STUDY ON THE APPLICATION OF NANOFIBRILLATED CELLULOSE
FROM PAPER INDUSTRY RESIDUES AS EXCIPIENT IN TABLETS
Keth Ribeiro Garcia
Universidade de Caxias do Sul (UCS)
Sara Elis Bianchi
Venina dos Santos
Valeria Weiss Angeli
Letícia Scherer Koester
Rosmary Nichele Brandalise
Introduction: Cellulose nanofibers (CNF) separation from fibrils of cellulose has aroused interest
because of its characteristics of high surface area, strength and stifness combined with high porosity,
biodegradability and renewability. CNF also shows potential to be used as excipient in pharmaceutical industry as
binders, tablets coating and as agents release modulators in the preparation of extended-release tablets. In order
to break down the cellulose structure, grinding and high-pressure homogenization process are usually employed to
obtain CNF. The aim of this paper is to obtain CNF by grinding in order to acquire a product for application in
pharmaceutical area in place of the most used microcrystalline cellulose (MC). Methods: Cellulose waste
(Eucalyptus sp.) from paper industry was dispersed in water (0.5 and 2%, w/v) and the suspension was fed in a
stone grinding mill adapted to a recirculation pump to promote fibrillation until a gel was formed. The gel obtained
was dried in a freeze-drying equipment. The freeze-dried (FD) sample was analyzed in a Field Emission Scanning
Electron Microscopy (FESEM) to evaluate the morphology of the fibers and that was compared with MC powder.
Results and discussion: The micrographs of FD, at both concentrations showed a net of individualized nanofibrils
with mean diameters of 67+-2nm and 40+-2nm, respectively, differently than MC that showed a diameter of
19.52+-3um. The difference between dimensions may be explained due to hydrogen bonds and van der Waals
forces that are expected to hold the fibrils together and form agglomerates after the removal of water. FD yields
were 1% and 2.05%, for the lower and higher concentration, respectively. Conclusions: The use of CNF, replacing
MC in tablet development has innovative character for drug matrix production. Hence, the biomass processing is
relatively simple; the design of pharmaceutical form takes into account principles of sustainability and indicates a
possible application
Financial Support
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES).
Acknowledgments
The authors are thankful to University of Caxias do Sul (UCS), to Laboratório de Microbiologia Aplicada (UCS), to Financiadora de Estudos e Projetos (FINEP)
(agreement 01.13.0359.00) and Laboratório Central de Microscopia Professor Israel Baumvol (UCS) for field emission scanning electron microscopy and to
Faculty of Pharmacy from Federal University of Rio Grande do Sul (UFRGS).
64
COMBINED EFFECTS OF NANOFIBROUS POLY-L-LACTIC ACID SCAFFOLDS AND
SIMVASTATINS ON DENTAL PULP STEM CELLS
Diana Gabriela de Sousa Soares
Department of Biologic and Materials Sciences, School of Dentistry, University of Michigan, Ann Arbor, MI, USA; Department of Physiology
and Pathology, Araraquara School of Dentistry/UNESP, Araraquara, SP, Brazil.
Zhanpeng Zhang
Department of Biologic and Materials Sciences, School of Dentistry, University of Michigan, Ann Arbor, MI, USA.
Fatma Mohamed
Carlos A. de Souza Costa
Department of Physiology and Pathology, Araraquara School of Dentistry/UNESP, Araraquara, SP, Brazil.
Peter X. Ma
Introduction: In the present study, the combination of a nanofibrous poly(l-lactic acid) (NF-PLLA)
scaffold and simvastatin, a lipid lowering molecule that has anabolic effects on osteoblastic precursors, was
evaluated for the odontogenic differentiation of human dental pulp stem cells (DPSCs) and dentin-like tissue
regeneration. Methods: The NF-PLLA scaffold was obtained by casting a solution of PLLA into interconnected
sugar spheres template (250 to 420 um diameter) under mild vacuum, followed by phase separation at -80oC and
freeze-drying. The DPSCs were seeded onto NF-PLLA scaffold and cultivated in osteogenic medium
supplemented with or without 0.1 uM/L simvastatin for two weeks. Then, the cell/scaffold constructs were
implanted subcutaneous into nude (nu/nu) mice. The samples were retrieved 8 weeks later, and stained with
hematoxylin-eosin (H&E), Masson's trichrome, Von Kossa and anti-DSP antibody. Results: The H&E staining
exhibited abundant cells in the macropores of the scaffolds in both groups. Vascular structures were formed inside
the scaffolds, with significantly higher vessel density in simvastatin group than in the control group. A greater
amount of collagen-rich matrix was found in the simvastatin group than in the control group. The
immunohistochemical analysis demonstrated significantly more intense DSP staining in the simvastatin group.
Finally, significantly higher mineral density was revealed in the simvastatin group. Conclusion; Simvastatin at a
low concentration enhanced the odontogenic differentiation of DPSCs and mineralized dentin-like tissue
regeneration.
Financial Support
CNPq # 303599/2014-6; FAPESP # 2014/23520-0; US NIH/NIDCR R01DE022327.
Ethical approval
The animal study was performed following a protocol (PRO00006076) approved by the Institutional Animal Care and Use Commit tee of the University of
Michigan, Ann Arbor, USA.
65
66
67
DEVELOPMENT OF A RP-LC METHOD FOR THE ANALYSIS OF RECOMBINANT
HUMAN INTERFERON BETA 1b
Alice Rosa da Silveira
Department of Industrial Pharmacy, Federal University of Santa Maria
Francielle Santos da Silva
Postgraduate Program in Pharmaceutical Sciences, Federal University of Santa Maria
Valquíria Guedes Perlin
Maurício Elesbão Walter
Douglas Franco Cardoso
Sérgio Luiz Dalmora
Introduction: Human interferons (IFNs) are natural proteins produced by the cells of the immune system
with antiviral, antiproliferative and immunomodulatory properties. Human interferon beta is a glycoprotein with 166
amino acids produced by fibroblasts. Interferons, IFNß-1a and IFNß-1b, are used as therapeutic agents.
Recombinant human interferon beta 1b, rhIFNß-1b, produced by DNA technology in E .coli consists of a 165
amino acids with molecular mass of 18.5 kDa and isoeletric point of 9.6, containing a genetically engineered
substitution of serine by cysteine at position 17. It is currently being used to treat multiple sclerosis, reduces the
relapse rate, relapse severity and progression of central nervous system. The aim of this study was to develop
and validate parameters of a reversed-phase liquid chromatography (RP-LC) method for the assessment of
rhIFNß-1b in biopharmaceutical formulations. Methods: The LC method was carried out on a Jupiter C4 column
(250 mm x 4.6 mm i.d.), maintained at 30°C. The mobile phase A consisted of 0.1% trifluoroacetic acid (TFA) in
water and mobile phase B consisted of 0.1% TFA in acetonitrile. The gradient elution was performed at a flow rate
of 1 mL/min with UV detection at 214 nm. The biological potency was evaluated using the bioassay based on the
antiproliferative activity of the A549 cells, reading the absorbances at 595 nm. Results and Discussion: The RPLC method demonstrated acceptable results for the accuracy, precision, specificity and robustness.
Biopharmaceutical batches of rhIFNß-1b were analysed by the RP-LC and by the in vitro bioassay, giving
potencies between 93.23 and 103.83% of the stated potency, with significant correlation (pD0.05). Conclusions:
The RP-LC method represents an alternative which can be used in combination with the bioassay, contributing to
improve the quality control and to assure the therapeutic efficacy of the biotechnology-derived product.
Financial Support
CAPES and FATEC
68
QUALITY PHYSICOCHEMICAL EVALUATION OF FUROSEMIDE TABLETS DISPENSED
IN A PUBLIC PHARMACY IN THE NORTHWEST OF RIO GRANDE DO SUL
Alisson Felipe de Oliveira
Universidade de Cruz Alta (UNICRUZ)
Gabriela Bonfanti
Regis Augusto Norbert Deuschle
Josiane Woultheres Botolotto
Viviane Cecilia Kessler Nunes Deuschle
Furosemide is classified as a benzenesulfonamide, acting on selective reabsorption of sodium chloride
in the ascending limb of Henle loop, as a loop diuretic. It is widely used in the treatment of hypertension, mainly
due to its fast action in acute cases. The quality control of medicines is important to prevent health risk, mainly in
Brazil public health network where medicines is purchased through bidding, when lower cost is taken into
account. Thus, the aim of this study was to evaluate the physicochemical quality of tablets content 40 mg of
furosemide, dispensed in a Public Pharmacy in northwest of Rio Grande do Sul. Therefore, the following tests
were performed in furosemide 40 mg tablets: average weight, friability and disintegration time, according the
procedures described in the Brazilian Pharmacopoeia, V edition. The results obtained in this study presented 168
mg ± 2.04 of average weight. The test has shown that the tablets have uniform weight, since the range of
variation allowed for tablets having weight within 80 mg and 250 mg is ± 7.5%. Thus, the lower and higher limits
are 155 mg and 180 mg, respectively, and each tablet unit tested in this study is within these established limits.
The total disintegration time for the tablets was 1 minute and 10 seconds, with no residue of the tested units after
this time. The result of the disintegration test is within the limits preconized by the Brazilian Pharmacopoeia,
which states that the tablets must be disintegrated within 30 minutes. The result from friability test was 0.26%,
again within the criteria preconized by the aforementioned Pharmacopoeia, where a maximum of 1.5% of weight
difference is allowed. The tablets analyzed follow the specifications, ie, the result reflects its capacity to
withstand mechanical stresses. In conclusion, all the results found in this study are within the established limits,
providing evidence of quality in furosemide tablets.
Financial Support
69
Qualitative analysis of substances with medicinal interest from stems and leaves of
specie Piper cernuum by gas chromatography coupled to mass spectrometer
Ana Cristina Gon
Pharmacy Graduate, University of Vale do Itajaí - UNIVALI, Itajaí, SC, Brazil.
Andressa Cristine Schaefer
Pharmacy Graduate, University of Vale do Itajaí - UNIVALI, Itajaí, SC, Brazil.
Tiago José Bonomini
Graduate program in Pharmaceutical Sciences and Center for Chemical-Pharmaceutical Research, University of Vale do Itajaí - UNIVALI,
Itajaí, SC, Brazil.
Theodoro Marcel Wagner
Tania Mari Belle Bresolin
Ruth Meri Lucinda da Silva
Angela Malheiros
The species Piper cernuum is one of the Piperaceae family species that has been evaluated for its
antimicrobial properties and disorders related to the central nervous system, but there are still few studies on the
phytochemical profile of the same. In this work it was aimed to perform qualitative analysis by gas
chromatography coupled with mass spectrometry (GC/MS) of the extracts of leaves and stems from the specie
Piper cernuum. The hydroalcoholic extracts (90 ºGL) of stems and leaves were obtained by static maceration for
7 days, filtrated and the solvent evaporated under reduce pressure in a rotary evaporator. Then the extracts were
submitted to a liquid-liquid partition, and the dichloromethane fraction was assessed by GC/MS (Shimadzu
GCMS-QP2010S). The identification of the constituents occurred by comparing the mass spectra with the NIST
library version 8.0. In stems it was identified bornyl coumarate (29.69%) as the major constituent, while in the
leaves this compound was not observed. The compound identified as dihydroagarofuran (24.55%) was the major
sesquiterpene found in the leaves, while in the stems were just 2.86%. In the leaves was possible to identify the
cubebin (7.76%), hinokinin (16.29%) and kusonokinin (1.86%), while in the stems were detected the cubebin
(2.46%), hinokinin (5.16%) and asarinin (3.74%). The bornyl ferulate (1.56%) was only detected in the stems. The
substances elemol, guaiol and beta-eudesmol, have been identified in both parts of the plant, but in different
concentrations. With this study can be concluded that the stems and leaves for Piper cernuum have variation in
chemical composition and concentration of secondary metabolites, for this reason for the continuation of studies
with this plant should be used stems and leaves separately, since this species has great potential as source of
bioactive molecules.
Financial Support
Scholarship Program of Scientific Initiation - PROBIC/UNIVALI
Acknowledgments
Public Call FAPESC/CNPq No. 06/2012 Support Program for Emerging Centers - PRONEM
70
VALIDATION AND APPLICATION OF SPECTROPHOTOMETRIC METHOD TO
DETERMINATION OF ATORVASTATIN CALCIUM IN TABLETS.
ANA JULIA PEREIRA SANTINHO GOMES
Federal University of São João del-Rei
Marina Goulart da Silva
CARLOS EDUARDO DE MATOS JENSEN
SABRINNA TASSIELE DA COSTA
Introduction. The atorvastatin calcium (ATC) is a drug used as a lipid-lowering agent. Today the ATC
has an exceptional dispensation, included in the specialized component dispensing program from the Sistema
Único de Saúde (SUS). It presents a relevant role on the treatment of patient who needs to reduce high LDL
cholesterol levels. The aim of this work was validate a spectrophotometric method, in UV absorption region, to
ATC and apply on tablets. Methods. This work describes the validation of an analytical method according to
Brazilian regulation (RE 899/2003, National Agency of Sanitary Vigilance) for determination of atorvastatin
calcium in tablets by spectrophotometry using phosphate buffer solution at pH 6.8. Three different samples of
ATC tablets were submitted at content uniformity (CU) and dissolution assays employing the developed method.
Results and Discussion. The results display that method is practical, selective, accurate, precise and linear
(r=0.9991) from 10 to 24 µg/mL of ATC measured at 242 nm. Moreover, the factorial planning indicated that the
method is robust in relation to temperature (7º or 25 ºC) and pH value (6.8 or 7.4). All samples presented ATC
dissolved at 30 minutes. CU results showed 97.3%, 102.4% and 108.1% of the labeled value, in mg, of ATC in
each tablet. However, the acceptance values found were 13.4, 7.08 and 15.85, respectively. Conclusions. The
results of content uniformity and in vitro dissolution assays, carried out with tablets, showed that the proposed
UV spectrophotometric method, previously validated, may be applied for ATC determination in pharmaceutical raw
material and solid dosage forms.
Financial Support
UFSJ
71
MICROBIOLOGICAL BIOASSAY BY AGAR DIFFUSION FOR DETERMINATION OF THE
ANTIFUNGAL POSACONAZOLE
Andressa da Silva Bitencourt
Laboratório de Controle de Qualidade Farmacêutico, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil
Sendy Sales Oliveira
Tércio Paschke Oppe
Andreas Sebastian Loureiro Mendez
Programa de Pós-Graduação em Ciências Farmacêuticas, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil
Cássia Virginia Garcia
Programa de Pós-Graduação em Ciências Farmacêuticas, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil
Introduction: Posaconazole is an extended-spectrum triazole antifungal agent for prevention and
treatment of invasive fungal infections. There are few reports of methods for its determination, especially when it
comes to bioanalytical, which justifies the importance of developing alternative methods for this finality. The
objective of this study was to develop and validate a microbiological bioassay by agar diffusion for determination
of posaconazole in raw material. Methods: The analytical conditions established were: Saccharomyces cerevisiae
ATCC MYA 1942 used as the test microorganism, methanol and pH 7.0 phosphate buffer as solvents, Sabouraud
Dextrose agar as culture medium, incubation at 30 °C for 48h, inoculum concentration of 2.0%. The
microbiological assay described in this study followed a 3×3 design. The method was appropriately validated by
determination of the parameters specificity, linearity, precision, accuracy and robustness. Results and
discussions: Preliminary tests using Candida albicans ATCC 10231 as test microorganism indicated that it was
not adequate to the conditions applied. In assessing the specificity, using the sample matrix, no significant
interference was found. For linearity, 9 calibration curves were obtained, with correlation coefficient of r = 0.999.
ANOVA showed the proposed method has significant regression. Precision was determined by repeatability (intraassay) and intermediate precision (inter-assay) and showed that values of relative standard deviation (RSD) were
below 5%, considered satisfactory. The mean amount found was 101.28%. Accuracy, determined by recovery
test, showed 101.61% of mean recovery. Robustness evaluation, performed through small modifications in the
analytical conditions, did not interfere with the results. Conclusions: Results showed that the proposed
microbiological method can be used as routine analysis for the quantitative determination of posaconazole in raw
material.
Financial Support
CNPq
72
DEVELOPMENT AND VALIDATION OF STABILITY INDICATION METHOD BY LC TO
DETERMINE LINAGLIPTIN IN THE PRESENCE OF POTENTIAL IMPURITIES
ANDRESSA TASSINARI DA SILVA
Laboratory of Research and Development in Quality Control, University Federal of Pampa - UNIPAMPA, Uruguaiana, RS, Brazil
GABRIELA ROSSI BRABO
RENATA MEDEIROS HILGERT
LISIANE BAJERSKI
CLESIO SOLDATELI PAIM
Postgraduate Program in Pharmaceutical Sciences, University Federal of Pampa - UNIPAMPA, Uruguaiana, RS, Brazil
Introduction. Bilastine is a novel non-sedating histamine H1-receptor antagonist developed for the
treatment of allergic rhino conjunctivitis and urticaria without show sedative or cardiotoxic effects in clinical trials
and in post-marketing experience. Although there are papers describing the determination of bilastine in
pharmacokinetic studies, there are no studies describing quantification methods to bilastine in tablets and no
information regarding the drug stability. Due to this, a stability indicating analytical method was developed and
validated by LC to determine bilastine in tablets. Methods. The chromatographic separation was performed in a
Shim-pack® RP-18 column (150 x 4.6 mm, 5 µm) using a Shimadzu Prominence® liquid chromatograph. The
mobile phase used was composed of a mixture of 0.3% triethylamine (pH 3.0) and acetonitrile (80:20,v/v) at a
flow-rate of 1.0 mL min-1. The injection volume was 10 µL and the run time was 5 min. Bilastine was determined
at 207 nm using photodiode-array detection. The developed chromatographic method was validated by evaluating
the following analytical parameters: specificity, linearity, precision, accuracy, and robustness. Furthermore, the
stability-indicating capability was determined by forced degradation conditions. Results and Discussion. Results
showed that there was no interference of the excipients and degradation products, obtained from stress
conditions, in the determination of the active pharmaceutical ingredient. The method showed suitable recovery
and intra- and inter-day precision (RSD < 2%). The response was linear over a range from 15.0 to 25.0 μg mL-1
(r= 0.9995) and the method showed robustness to small modifications in the analytical conditions. Conclusion.
The results confirm that the proposed method can be used for quantitative determination and to verify the stability
of this drug.
Financial Support
PROJETO UNIVERSAL CNPQ (Processo 457629/2014-2), PDA UNIPAMPA
Acknowledgments
CNPq and UNIPAMPA
73
DEVELOPMENT AND VALIDATION A STABILITY INDICATING METHOD FOR
SIMULTANEOUS DETERMINATION OF IBANDRONATE AND ITS IMPURITIES BY HIGH
PERFORMANCE ION EXCHANGE CHROMATOGRAPHY AND CHARGED AEROSOL
DETECTOR.
Bianca Gonzalez Fernandes Machado
Lúcio Mendes Cabral
Valéria Pereira de Sousa
Introduction: High performance liquid chromatography method can be especially challenging for very
polar compounds, such as ibandronate and its related impurities, which both are poorly retained on reverse-phase
columns and lack a sufficient UV chromophores or fluorophore. The aim of this present study was to develop and
validate a stability-indicating isocratic anion-exchange chromatography method for the quantitative analysis of
ibandronate in the presence of its main impurities generated from forced degradation studies using a charged
aerosol detector. Methods: Separation was achieved with an aqueous mobile phase composed of formic acid at
the flow rate 0.8mL.min-1. CAD was performed using a nitrogen pressure of 35 psi and a range of 100 pA.
Validation parameters such as specificity, linearity, limit of detection and quantification, precision, accuracy and
robustness were analyzed. Results and Discussion: The specificity and stability-indicating of the method were
evaluated though the forced degradation studies. The drug was subject to acid and alkaline hydrolytic conditions,
oxidative and metal decomposition, thermal and photolytic conditions. The drug degrades under oxidative and
metal conditions, but was stable in the other conditions. Degradation products resulting from the stress studies
did not interfere with the detection of ibandronate. The retention time of the drug was approximately 5 min. The
resolution of the drug and degradants peaks was > 2.0. The limit of quantification values were found to be 25.165;
0.889; 0.421 and 2.377 µg.mL-1 for ibandronate, phosphate, phosphite and 3-methylpentylaminopropionic acid,
respectively. Conclusions: The method was validated with respect to linearity, precision, accuracy, selectivity,
specificity and robustness. Thus, the proposed method was simple, rapid, selective, accurate, stability indicating
and suitable for routine quality control of ibandronate in drug substance or in its dosage form.
Financial Support
FAPERJ, CAPES and CNPq.
74
COMPARISON OF THE HPLC-UV AND UFLC-DAD TECHNIQUES TO DETERMINATION
OF COUMARINS OF Pterocaulon balansae IN AQUEOUS EXTRACT
Bruna Medeiros-Neves
Marina Cardoso Nemitz
Pharmaceutical Sciences Graduate Program, University of Rio Grande do Sul, Porto Alegre, RS, Brazi
Helder Ferreira Teixeira
Pharmaceutical Sciences Graduate Program, University of Rio Grande do Sul, Porto Alegre, RS, Brazi
Introduction: The literature described different biological activities of coumarin-rich extracts of
Pterocaulon (Astereceae). In this context, the aim of this study was to develop and validate a method a high
performance liquid chromatography (HPLC-UV) and an ultra fast liquid chromatography (UFLC-DAD) for the
determination of coumarins of Pterocaulon balansae in aqueous extract. Methods: Chromatographic separation
was performed on a Phenomenex-C18 RP column for HPLC-UV and Shim-pack XR ODS column for UPLCDAD. The mobile phase consisted of 0.1% formic acid (A) and acetonitrile (B) using gradient elution, flow rate and
column oven was HPLC (1 mL/min; 30ºC) and UFLC (0.55 mL/min; 55ºC) for 45 min and 8 min, respectively. The
linearity curve used as standard 5-methoxy-6,7-methylenedioxycoumarin (5MMDC) in a concentration range from
0.1 to 7.5 μg/mL. The accuracy was evaluated at three levels: 0.5, 2.5, and 5.0 μg/mL for 5MMDC. The mean,
standard deviation, RSD and 95% confidence interval for all concentration levels were determined for the three
days. Results and Discussion: The coefficient of determination (r2) obtained was 0.9999 for both methods, which
demonstrates the high correlation between the concentration of 5MMDC and peak area. The LOD and LOQ
demonstrating a satisfactory sensitivity for the both methods. The recoveries were 99.31 - 106.11%, without
statistical difference and RSD < 2.0% for all determinations. These methods showed good accuracy at the three
concentrations, within the linearity of the method. The results showed RSD lower than 2% for experiments on the
same day or three different days, indicating that these methods were precise, according to the official codes.
Conclusions: The methodologies proposed proved to be linear, precise, and accurate to estimate coumarins from
aqueous extracts of P. balansae, without significant differences between methods.
Financial Support
CNPq/Brazil and CAPES/Brazil.
75
DEVELOPMENT CHROMATOGRAPHIC METHOD OF ANALYTICAL ORGANIC
IMPURITIES OF SITAGLIPTIN
Camila Ferrazza Alves Giordani
Sarah Campanharo
Elfrides Eva Sherman Shapoval
Martin Steppe
Introduction: The control and evaluation of impurities in the pharmaceutical industry are critical to the
quality, safety and efficacy of drugs developed. Regulatory authorities have incorporated the limits with regard to
permissible levels of impurities present in the raw material and the finished product where it's necessary to
correct selection of analytical methods with sufficient sensitivity to measure small amounts of impurities. Among
the diseases that require continuous use drugs, there is the type 2 diabetes mellitus being the class of gliptins
features rare studies for the assessment and quantification of impurities. Until now, there isn't any research of
impurity synthesis of sitagliptin. Materials and Methods: The reference standard and impurities were purchased
from Sequoia Research Products and Sigma Aldrich. The LC method was carried out on na HPLC Agilent® liquid
chromatograph. The UV detector was operated at 207 nm. Results and Discussion: in order to obtain the best
chromatographic conditions, several parameters were tested based in previous analysis. Different reversed-phase
columns, mobile phase composition, pH values and isocratic mobile phase systems were used in order to
separate sitagliptin, impurity 1 and impurity 2. With the intent to elute them as symetrical peaks, the optimization
of parameters was performed based on resolutions among drug and impurities, asymetric factor and theoretical
plates. The retention time observed were 3.86, 4.89 and 7.35 for impurity 1, 2 and sitagliptin, respectivelly. The
resolution were 6.33, 4.91 and 8.96, the asymetric factor were 0.73, 0.72 and 0.88, the theoretical plates were
7729, 8969 and 6366 for the impurity 1, 2 and sitagliptin, respectivelly. Conclusions: The method development
showed good results for the simultaneous analysis of sitagliptin and their impurities synthesis and have prospects
for conduct the studies about validation of analytical methodology.
Financial Support
Financial support from CNPq/Brazil and master scholarship from CAPES/Brazil.
76
PHOTOSTABILITY AND DEGRADATION KINETICS OF ANTIFUNGAL POSACONAZOLE
IN RAW MATERIAL
Caren Gobetti
Andressa da Silva Bitencourt
Sendy Sales Oliveira
Vanise Coty Rodrigues
Introduction: Posaconazole is an antifungal drug belonging to triazole class, characterized by a wide
range of action, being indicated for the prophylaxis of invasive infections from Aspergillus spp. and Candida spp.
Few reports about drug stability are available, mainly exploring the behavior on degradation kinetics. The present
study purposed to establish the effect of UVC photoradiation on the decomposition of this antifungal in raw
material, determining the degradation kinetics by HPLC and microbiological assay. Methods: HPLC method was
performed as described in an earlier publication, whose validation was described. Microbiological analysis was
conducted applying agar diffusion-cylinder-plate assay, previously validated on concentration range of 2.5 - 10.0
µg/ml. Photostability study was carried out through employment of stress conditions for different time periods (0,
15, 30, 45, 60, 90, 120 and 180 minutes). Kinetics decomposition was determined by plotting quantitative data on
graphical performance and observing the linearity tendency. Results and Discussion: Approximately 80% of drug
degradation was observed after exposure for 180 minutes. Two degradation product peaks were found at 7.0 and
9.1 min (drug retention time = 7.8 min). The degradation rate kinetics, determined by plotting log of drug
concentration remaining versus time, could be described by first-order profile. The drug content was
simultaneously determined by microbiological assay, which indicated that posaconazole decomposition follows a
second-order reaction model. This different kinetics profile can be justified by particularities intrinsic to described
bioassay. Conclusions: Posaconazole demonstrated to be very sensitive to UVC radiation. Despite of equivalent
on potency determination, the bioassay must be better evaluated when kinetics is studied. HPLC method is a
reference for stability profile, since involves a refined monitoring focused on drug peak, without interferences.
Acknowledgments
CNPq; BICPROPESQ (UFRGS)
77
CHROMATOGRAHIC METHOD DEVELOPMENT FOR PHOTOSTABILITY EVALUATION
OF ANTI-INFLAMATORIES-LOADED NANOCAPSULES
Carlos Eduardo da Rosa Silva
KELLY AYUMI NAKAMA
SIMONE PINTON
SANDRA ELISA HAAS
Introduction: In the treatment of Alzheimer's disease, the therapy based on anti-inflammatory is
emergent, and the association of Curcumin (CM) and Meloxicam (MX) in nanocapsules (NC) is innovative and an
interesting strategy to improve their solubility, permeability, bioavailability and consequently their efficacy. We
intend to develop an HPLC-PDA method able to quantify CUR and MLX simultaneously aiming to determine the
photostability (PS) of both drugs free and nanoencapsulated. Methods: NC were prepared by nanoprecipitation
method. Encapsulation rate (ER) was determined through the ultrafiltration/ultracentrifugation technique using
Ultrafree® units. Drug loading (DL) was obtained from dissolution of NC in methanol. The PS was evaluated
using a photostability chamber with 2 UV lamps. Samples were collected, diluted in methanol, filtered (0.45 um)
and injected into HPLC. HPLC-PDA (Shimadzu) was used with a C18-RP column, triethylamine
water/methanol/acetonitrile as mobile. The validation procedures were performed according to ICH guidelines and
RE nº899/2004-ANVISA. Results and Discussion: Method was validated according to Linearity r>0.999, precision
with maximum RSD of 1.78%, accuracy with recovery between 98.44 and 101.66%, specificity showed not
interference, peak purity was 1.00, robustness with small variations in mobile phase pH and flow rate the results
obtained are between 98.20% and 104.77%. The ER were 100.01 % and 99.88% for CM and MX respectively,
and the DL was 94.43% and 95.56% for CM and MX. In the PS we obtained a degradation of 47.65 for MX and
55.38% for CM in solution comparing with 31.38% and 38.84% when drugs were entrapped into NC. No
interference was observed in the chromatograms during analysis. Conclusion: The method was developed and
validated for CM and MX quantification and it is applicable for a PS study. The ER and DL were close to 100% for
both drugs and NC can protect the compounds of photodegradation.
Financial Support
CNPQ/Brazil; FAPERGS/Brazil
Acknowledgments
Science Pos-Graduate Program
78
A new assay method for calcium carbonate tablets by using Thermal Infrared
Enthalpimetry
Cristiane Franco Codevilla
Universidade Federal de Santa Maria (UFSM)
Bruna Tischer
Alessandra Stangherlin Oliveira
Daniele de Freitas Ferreira
Sandra Kunde Schlesner
Flávia Michelon Dalla Nora
Bruna Nichelle Lucas
Cristiano Ragagnin de Menezes
Roger Wagner
Juliano Smanioto Barin
The enthalpimetry is a technique based on quantitative determination of a reactant or catalyst through
the enthalpy change of a chemical reaction. In this study, the infrared thermal imaging was used as a tool for
measurement of temperature change of several reactions simultaneously. Therefore, the potentiality of thermal
infrared enthalpimetry (TIE) was demonstrated for the determination of calcium followed by comparison with the
conventional technique. The TIE was performed in 24-well microplates using multichannel pipette and an infrared
camera for monitoring the temperature released inside the wells after addition of the reagents. For the
determination of calcium by TIE 30 tablets of calcium carbonate were triturated, homogenized and the solution
was adjusted at pH 9. For construction of the calibration curve used was calcium chloride dihydrate (0.1 to 1.0
mol L- 1) as reference solutions and tetrasodium EDTA excess (0.6 mol L-1) as reagent in excess. In this way
1200 µL of reference solution or sample were inserted into each well and 1200 µL of tetrasodium EDTA was
added. For determination with conventional method, titration with 0.05 mol L-1 disodium EDTA solution, around
0.1 g of the sample was weighed and diluted in 50 mL of water plus 2 mL of HCl (0.1 mol L-1), with blue hydroxyl
naphthol as indicator. The calibration curve obtained by TIE showed an excellent coefficient and determination
(R2 = 0.9978) and the proposed method found calcium content (in mg of CaCO3 / tablet) of 1281 ± 18 by TIE and
1268 ± 22 by conventional technique with no significant differences between them. The results showed good
agreement with conventional techniques, making analysis quickly, effectively and with the potential to be applied
to routine analysis, due to the simplicity, speed and high frequency analysis. Thus, it was shown in this work that
the TIE may be considered an important tool for the determination of analytes often determined by titration.
Financial Support
Capes
79
SPECTROSCOPIC, MORPHOLOGICAL AND CRYSTALLINE EVALUATION OF SOLID
DISPERSED SYSTEMS WITH 6CN AND SOLUPLUS
Danielle Lima Bezerra de Menezes
Walter Ferreira da Silva Junior
Nathália Ribeiro Calazans Brandão da Silva
Zênia Maria Maciel Lavra
Francisco Jaime Mendonça Bezerra Júnior
Universidade Estadual da Paraíba (UEPB)
Introduction: 6CN is a new derivative of 2-amino thiophene with indicative anti-inflammatory activity, but
has low solubility in water. The use of hydrophilic polymers to increase solubility of hydrophobic drugs in solid
dispersion (SD) has been widely used, being Soluplus a viable alternative due to its amphiphilic and not ionic
properties. This study aimed to get SD with hydrophilic copolymer Soluplus characterized by Fourier transform
infrared spectroscopy (FTIR), X-ray diffraction (XRD) and scanning electron microscopy (SEM). Methods:
Physical mixtures (PM) and the SD have been developed by kneading (KN) and rotary evaporation (EV) methods
in a 1:1 w/w ratio. Results and Discussion: The 6CN spectra has shown characteristic bands at 2194 cm-1 region
of axial deformation of the C≡N group, and two bands about a 3320 and 3440 cm-1 corresponding to the axial
deformation N─H. There are bands in 1614 cm-1 related to C═C bond and in 1517 cm1 to the C─H connection.
The spectra were similar to 6CN, compared with PM. Both KN and EV has shown a decrease in the intensity of
the characteristi bands, but the EV of 6CN showed narrowing of the bands indicating a sign of greater interaction.
The micrographs have shown the 6CN crystals of well-defined ways and Soluplus is characterized by irregular,
porous, spherical and amorphous particles. In the PM it is still possible to observe 6CN between the polymer
crystals, but in the KN and EV the crystals have disappeared, indicating the dispersion on matrix carrier. The
diffractogram of 6CN has shown a crystallyne profile, though Soluplus have been amorphous profile. All systems
show a reduction in the crystalline profile 6CN, most evidently in EV. Conclusions: With these results, it is
possible to suggest that the DS obtained by rotary evaporator with Soluplus is na alternative for increasing the
solubility of prototype.
Financial Support
This study has financial support from CNPq.
80
(2E) -1-phenyl-3- (4-methoxy-phenyl) -2-propen-1-one CHALCONE: SYNTHESIS,
CHEMICAL, PHYSICOCHEMICAL CHARACTERIZATION AND BIOCOMPATIBILITY
Danyela Francine Benvenutti
Pharmaceutical Sciences Graduate Program, Universidade do Vale do Itajaí, Itajaí, SC, Brasil
Angelica Garcia Couto
Pharmaceutical Sciences Graduate Program, Universidade do Vale do Itajaí, Itajaí, SC, Brasil; NIQFAR, CCS, Universidade do Vale do
Itajaí, Itajaí, SC, Brasil;
Maurizio Ricci
Dep. of Pharmaceutical Sciences, University of Perugia, Perugia, Umbria, Italy
Serena Casagrande
Giada Ceccarelli
Fatima de Campos Buzzi
Valdir Cechinel Filho
Rogério Corrêa
Tania Mari Bellé Bresolin
Introduction: The chalcone (2E) -1-phenyl-3- (4-methoxy-phenyl) -2-propen-1-one is a promissory
molecule, due to its anti-inflammatory action and osteogenic activity However, there is no information about its
solubility, polarity, stability or toxicity, whose data are relevant to the establishment of a new drug and to the
development of a rational delivery system. Methods: The synthesis of chalcone was performed according to
Claisen-Schmidts method. The reaction was monitored by TLC. The chalcone was purified by column
chromatography, monitored by TLC and recrystallization. Chemical characterization was performed using CG-MS
and FTIR. The solubility was evaluated according to pharmacopeial method, using different solvents like DMSO,
acetonitrile, methyl alcohol, ethyl alcohol, dichloromethane, PBS buffer and water. The thermodynamic solubility
was evaluated as well. The thermal behavior was analyzed (DSC and TG), and also the toxicity effect on Chick
Chorioallantoic Membrane (CAM) assay was performed at 1 and 10 µM of chalcone. Results: The CG-MS of
synthesized chalcone showed a high purity, with the molecular ion at m/z 238, base peak at m/z 77 and the peak
at m/z 161, referring to fragments resulting from the breakage of the carbon. The FTIR showed absorption bands
characterized according to literature. DSC curve showed an endothermic event with Tonset at 74,9 ºC,
characteristic of the melting process of the substance. The decomposition process begins in temperature 196,8
ºC, with mass loss of 88.26%. The purity, by equation Van't Hoff, was of 100.2 mol%. The compound was
practically insoluble in water, slightly soluble in ethanol and methanol and easily soluble in DMSO and
dichloromethane. The preliminary CAM results showed no toxicity up to 10 µM of chalcone, after 3 days of
incubation. Conclusion: The chalcone was successfully synthetized with high purity, proving to be a hydrophobic
and relatively biocompatible compound.
Financial Support
Capes
Acknowledgments
The authors thank CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior), Brazil, for providing financial support in the form of a doctoral¿s
degree scholarship to Benvenutti D.F. and to SCIENCE PROGRAM WITHOUT BORDERS - SCHOLARSHIPS IN THE COUNTRY, TYPE RESEARCHER SPECIAL
VISITOR ¿ PVE (process n. 88881.064960/2014-01)
81
NMR QUANTIFICATION IN COMPLEX MIXTURES - OSMAC METHODOLOGY TO
INCREASE METABOLITE PRODUCTION IN Fusarium oxysporum EXTRACT ISOLATED
FROM Senna spectabilis's RIZHOSPHERE
Denise Medeiros Selegato
Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP)
Rafael Teixeira Freire
Universidade de São Paulo
Ian Castro-Gamboa
Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP)
Introduction F. oxysporum, is majorly isolated from S. spectabilis's rizhosphere and produces several
bioactive metabolites. Among those, fusaric acid is abundantly produced and plays an important role on microbe's
protection. Furthermore, it also displays antimycobacterial activity and potent inhibition of dopamine-B-hydrolase
in humans. Traditional culture techniques restrict metabolic pathways, silencing genes responsible for metabolite
production. In this sense, one strain, many compounds (OSMAC) emerges as a reliable approach for activing
these pathways, usually by altering cultivation parameters in a design of experiment. This work aims to optimize
fusaric acid production by altering cultivation variables in a factorial planning that uses NMR quantification as
response. Additionally, this work provides accurate mycotoxin measurement in crude extractas, as well as
dereplication of other derivates by LC/HRMS/MS. Method Complete factorial planning evaluated three variables of
cultivation, each tested on two different levels, totalling 8 experiments. Variables tested were (a) temp. 28C (+1);
14C (1); (b) agitation at 120 rpm (+1) and static (1); (c) presence (+1) or absence of light (-1). Ethyl acetate
extracts were analysed by 1H-NMR for quantification. Results/Discussion qNMR displayed different fusaric acid
production, yielding 11.89 mg (exp.1), 36.18 mg (exp2), 15.91 mg (exp.3), 42.73 (exp.4), 1.25 mg (exp.5), 1.22 mg
(exp.6), 2.15 mg (exp.7), 1.73 mg (exp.8). Factorial planning showed that agitation and temperature were the most
significant variables and the production reaches its highest at 28C and 120rpm, producing up to 42% of fusaric
acid. Conclusion Factorial planning successfully enabled the maximization of fusaric acid production by qNMR in
crude extract and, through fragmentation analysis, it was possible to further dereplicate two derivates (1) 5propylpyridine-2-carboxylic acid and (2) 5-(3-buten-1-yl)-pyridine-2-carboxylic.
Financial Support
This work was funded by grants from Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP): a Ph.D scholarship awarded to D.M.S (2014/059350), the Biota-FAPESP Program (2011/50816-1) and, CEPID Program though CIBFar Project (2013/07600-3).
Acknowledgments
Additional research grant awarded to I.C-G (449523/2014-4) funded by Conselho Nacional de Desenvolvimento Científico e Tecnológico ¿ CNPq is also
acknowledged.
Ethical approval
N/A
82
NANOCAPSULES CONTAINING CHIA OIL FOR WOUND CARE .
ELENICE SPAGNOLO RODRIGUES MARTINS
Centro Universitário Franciscano- Santa Maria- RS
Cayane Genro Santos
Isabel Roggia
Solange Cristina da Silva
INTRODUCTION: The wound healing is a process that can be accelerated through the use of products
such as Essential Fatty Acids (EFA). The chia (Salvia hispanica L.), originating in Mexico and Colombia, has
been used in skin lesion treatments because their oil is rich in EFA, such as linoleic acid (LA) and linolenic acid
(ALA). AL plays an important role for macrophage chemotactic, promoting autolytic debridement of the wound,
inducing granulation and consequently their healing. The nanocapsules enhance the stability of drugs, be for the
degradation by pH, the action of light, oxygen or other components of the formulations. This study aimed to
develop, characterize and evaluate the stability of nanocapsules with Pluronic® F127, containing chia oil for the
treatment of wounds. METHODOLOGY: The preparation of the nanocapsules was performed according to
Esposito et al (2012) with some modifications. The components of the organic phase were kept under magnetic
stirring at a temperature of 60 ° C until complete dilution of the components. The suspension with a final volume
of 50 ml was placed in the Rotary evaporator to the elimination of the organic solvent. The size, polydispersity
index and zeta potential of the nanocapsules were verified by dynamic light scattering using equipment
Zetasizer® nano-ZS model ZEN 3600, Malvern®. RESULTS:The nanocapsules containing chia oil, were
assessed over a period of 28 days for a 7-day interval where the sample was kept in the refrigerator. The size of
the nanocapsules was less than 260 nm, the polydispersity index is less than 0.3 and Zeta potential varied
between -4.0 and -5.0 mV. CONCLUSIONS: The results indicate that the nanocapsules containing chia oil,
encapsulated with Pluronic® F127, showed a desirable size, polydispersity within the parameters after analyzing
a period of 28 days.
83
IN VITRO-DISSOLUTION PROFILE OF ZIPRASIDONE HYDROCHLORIDE CAPSULES:
COMPARISON BETWEEN TRADITIONAL ANALYSIS versus MODELING
Fabricia Ré
State University of Maringá - Depart. of Pharmacy
Bruna Luiza Pelegrini
Fábio Pinheiro de Souza
Prati-donaduzzi - Pharmaceutical Industry
Andréa Diniz
Josmar Mazuchelli
State University of Maringá - Depart. of Statistics
Marli Mirian de Sousa Lima
In vitro-dissolution tests provides essential information for drug development. Ziprasidone, an
antipsychotic drug, is largely used in Brazil. To build a better knowledge about this drug behavior, the aim of this
research was characterize the dissolution profile of ziprasidone by traditional dissolution and flow cell apparatus.
The dissolution profile of reference (Geodon®) and test was conducted in apparatus 2 (1000 mL of medium, 5
rpm) and flow cell (apparatus 4) (open system, 22.6 mm cells, flow 12 mL/min, pH gradient). Five media were
used on apparatus 2 (Simulated Gastric Fluid (SGF) + 1 or 0.5% sodium lauril sulphate (SLS), Sodium acetate
buffer (SAB) pH 1.2 and 6.5 (plus 0.5% SLS) and Feed fluid simulated (FeSSIF) pH 5.0. The dissolution profiles
were compared by independent models (F1, F2, dissolution efficiency - ED%) and by dependent models methods.
The F2 detected similarity between the profiles in all the experimental conditions, but F1 not approved the profile
comparison to FeSSIF pH 5,0. The dissolution efficiency showed statistically significant differences in gastric
fluid and apparatus 4 profile. The model Korsmeyer-Peppas showed acceptable adjustments and indicated that
the release mechanism involved in all experimental conditions in apparatus 2 is Fick diffusion process. The
presence of residual water in the pregelatinized starch and the low solubility of ziprasidone can attribute the profile
as pseudo-matrix. The Gompertz and logistic models presented acceptable adjustments at the profile apparatus 4,
however, kinetic meaning of these two models is unclear. Additional statistical approaches showed differences
between two tested formulations, which were not detected by the method currently required by regulatory
agencies. The study of in vitro- dissolution associated to accurate statistical analysis showed an interesting
approach to guide drug development.
Financial Support
CNPq, MCTI/FINEP/CT-INFRA.
Acknowledgments
Prati-donaduzzi - Pharmaceutical Industry.
84
PHOTOLYTIC DEGRADATION OF FLUOROQUINOLONES: COMPARISON OF
DIFFERENT STRUCTURES AND IN SILICO STUDIES
Fávero Reisdorfer Paula
Universidade Federal do Pampa
André Valle de Bairros
Pharmaceutical Sciences Post Graduate Program, Federal University of Pampa, Uruguaiana, RS, Brazil
Danillo Baptista Pereira
Course of Pharmacy, Federal University of Pampa, Uruguaiana, RS, Brazil
Everson Willian Fialho Cordeiro
Clésio Soldateli Paim
Fabiana Ernestina Barcellos da Silva
Pharmaceutical Pharmaceutical Sciences Post Graduate Program, Federal University of Pampa, Uruguaiana, RS, Brazil
Marcelo Donadel Malesuik
Introduction: Fluoroquinolones (FQs) are antibacterial agents widely used around the world. These
compounds may present instability in light environment and degradation products generated may show some
adverse effects according the distinct chemical structures. These compounds present differences in the behavior
of structure degradation, such as paths of degradation or photochemistry reactions, when exposed to UV light.
This fact may indicate the necessity of understanding the differences among the FQs reactivity. In this work, the
photolytic hydrolysis of four FQs, such as ciprofloxacin, danofloxacin, gemifloxacin, and norfloxacin, and
molecular modeling of chemical structures were studied aiming to observe the differences among molecular
reactivity. Methods: Forced degradation was performed using a chamber UVC (254 nm) at 5-30 minutes using
diluted compounds in 100 µg/mL. All degradation was analyzed by HPLC-DAD, with reverse phase column C-18
(250 mm x 4.6 mm x 5 µm ID), using pharmacopeial and validated methods. In silico studies the software
Spartan'08 for Windows and DFT/B3LYP-6.31G* method were used to perform the FQs geometry optimization.
The physicochemical properties (Energies of HOMO, LUMO and GAP), generation of space filling and maps of
EHOMO and E LUMO were calculated and also used to assist the understanding the structure chemical
degradation. Results and discussion: Gemifloxacin and danofloxacin, showed more degradation perceptual
indices in comparison with ciprofloxacin and norfloxacin. Norfloxacin is the more stable compound than other
three FQs. It was observed some structural features which may be influencing on degradation, such as the
presence of ring of five members attached in quinolone ring and physicochemical properties as EHOMO.
Conclusion: Gemifloxacin was more instable compound in photolytic conditions analyzed. Structural features and
physicochemical properties studied may be used in the research of effective and more stable FQs.
Financial Support
CAPES (PNPD 2014), CNPq, FAPERGS, PDA-UNIPAMPA 2015
Acknowledgments
PPGCF-UNIPAMPA
85
PHYSICOCHEMICAL CHARACTERIZATION OF SIMVASTATIM-CHITOSAN FILMS
FELLIPE FERNANDES SANTOS
VANDIARA MARTINS MOREIRA
AMARO CÉSAR LIMA DE ASSIS
NATAN EMANUEL SOBRAL DE SILVA
PEDRO GOMES SILVEIRA NETO
YURI BASILIO GOMES PATRIOTA
MALU MARIA LUCAS DOS REIS
BRUNA PEREIRA DA SILVA
MYSRAYN YARGO DE FREITAS ARAUJO REIS
BOLÍVAR PONCIANO GOULART DE LIMA DAMASCENO
Characterization of new drug delivery systems became of extreme importance, since the greater the
amount of information obtained on a system, the easier it becomes to understand its mechanisms such as
solubility increase, crystalline modification and product stability. The aim of this study was to utilize techniques
such as Fourier transformed infrared (FTIR) and x-ray diffraction (XRD) to characterize simvastatin (SINV)chitosan (CTS) films. An FTIR analyses utilized a 4000 to 650 cm-1 scan and for the XRD analysis an angular
scan of 5° < 2θ < 60° was performed. The FTIR films demonstrated SINV bond suppression, which suggested
the drug incorporation. This kind of system arrangement can occur through the formation of non-covalent bonds
such as hydrogen bond and van der Waals forces. Moreover, there is a possibility of the SINV forming
intermolecular hydrogen bonds with CTS, what it leaves its hydroxyls free and able to forming bonds with water,
which result in an increase in aqueous solubility. The CTS film XRD analysis presented crystalline reflections
starting between 9° and 10° and 19° and 20°, which is characteristic of pure CTS diffractorgrams, what indicate
drug protection by the polymeric matrix. In the SINV-CTS film, physicochemical interactions occurred, which
caused the loss of the drug structure crystallinity, consequently increasing its aqueous solubility, which agreed
with the FTIR assay. Since it is less thermodynamically stable, the drug amorphous form is readily released.
Therefore, it can be inferred that the SINV crystalline modification could improve it aqueous solubility, which
supports its incorporation into the polymeric film and drug release from the polymeric matrix.
Acknowledgments
CAPES, CNPq, CERTBIO.
86
EVALUATION OF DIFFERENT MATRICES FOR 3D ORGANOTYPIC ORAL MUCOSA
MODEL
Fernanda Gonçalves Basso
Araraquara School of Dentistry - UNESP
Taisa Nogueira Pansani
Cynthia Marcelo
University of Michigan
Stephen Feinberg
University of Michigan
Josimeri Hebling
Carlos Alberto de Souza Costa
Introduction: Three-dimensional (3D) cell culture models may better simulate in vivo conditions for cells
interactions and phenotypes. The main objective of this study was to compare three different scaffolds for the
fabrication of a cellular 3D connective tissue model with gingival fibroblasts for development of a full thickness in
vitro oral mucosa model. Methods: Three different dermal matrices were selected: acellular cadaveric human
dermis (AlloDerm®); acellular porcine dermis (Strattice®); and collagen type I. Human oral keratinocytes and
gingival fibroblasts were obtained by enzymatic digestion using trypsin 0.04% and collagenase type II,
respectively. Gingival fibroblasts were seeded (1x105 cells/matrix) on the dermal side of the AlloDerm® and
Strattice® or within the type I collagen matrices using complete culture medium (DMEM), which was replaced
every 24 hours. After 72 hours, the matrices were individually transferred to transwell inserts and oral
keratinocytes were seeded onto them (1x105 cells/ matrix). Matrices were kept submerged in EpiLife culture
medium supplemented with epidermal growth factor (EGF) for 4 days. Then, samples were transferred to an airliquid interface for additional 7 days. Results and Discussion: Fibroblasts were able to adhere to the dermal
surface of all dermal matrices; however, poor migration of cells was observed within them. Higher fibroblast ratio
was observed when these cells were seeded into the type I collagen matrix, in which the distribution of gingival
fibroblasts was seen to be more homogeneous. Epithelial cells were able to adhere and proliferate on all three
matrice's surfaces; however, higher stratification and differentiation was obtained only for the two dermal
matrices. Conclusion: Type I collagen matrix provides a better environment for fibroblasts and epithelial cells
survival and proliferation, thus providing a more suitable model for fabrication of a full 3D oral mucosa equivalent.
Financial Support
São Paulo Research Foundation (FAPESP: # 2012/179478; 2013/058790; 2014/060577); National Counsel of Technological and Sci entific Development (#
442637/201404; 157779/20157).
Acknowledgments
the LifeCell Corporation for the donation of matrices, the São Paulo Research Foundation and the National Council for Scientific and Technological Development.
Ethical approval
Cells were isolated according to protocol approved by the Ethics Committee of University of
Michigan and Araraquara Dental School
87
VALIDATION OF A SIMPLE AND FAST UV METHOD FOR QUANTIFICATION OF
MANIDIPINE DIHYDROCHLORIDE FROM PELLETS APPLIED TO IN VITRO
DISSOLUTION AND STABILITY ASSAYS
Fernanda Malaquias Barboza
Pharmaceutical Sciences Graduate Program, Department of Pharmaceutical Sciences, State University of Ponta Grossa, Ponta Grossa, PR,
Brazil.
Vanderson Galan
Pharmaceutical Sciences Graduate Program, Department of Pharmaceutical Sciences, State University of Ponta Grossa, Ponta Grossa, PR,
Brazil.
Fabio Pinheiro De Souza
Prati-Donaduzzi Pharmaceutical Company, Toledo, PR, Brazil.
Sarah Moherdaui Martins
Vinícius Müller
Jessica Mendes Nadal
Department of Pharmaceutical Sciences, State University of Ponta Grossa, Ponta Grossa, PR, Brazil.
Paulo Vitor Farago
Department of Pharmaceutical Sciences, State University of Ponta Grossa, Ponta Grossa, PR, Brazil.
Validation is essential to define if a developed method is completely adjusted to the intended objectives,
in order to get trustworthy results that can be satisfactorily interpreted. The aim of this study was to develop and
validate a method to determinate manidipine in controlled release pellets innovative formulations by UV-VIS
spectrophotometry method. Two different routes for obtaining pellets containing manidipine have been proposed,
the first was used coating technique in the fluidized bed for both the production of the active pellets and functional
layer. The second path containing the active pellets were prepared by extrusion-spheronization route wet
granulation and subsequent coating steps were done in a fluidized bed. The proposed method was validated as
per the ICH guidelines and Resolution Nº 899 of 2003 (Brazil). The method was validated for specificity, linearity,
accuracy, precision and robustness. Both formulations prepared were successful obtained, however, that one
produced by extrusion-spheronization technique presented lower yield rates. This UV method was found to be
specific, precise, accurate, and robust besides being fast and simple and cheap. The UV scanning showed that
the excipients did not interfere with the analysis. The correlation coefficient (0.9999) indicated an excellent
linearity which allows establishing relations between absorbance and manidipine concentration. Regarding
precision, all tests presented a standard deviation lower than 5.0%. The method showed accuracy when
demonstrated a great proximity between sample solution and nominal concentration. Furthermore, the low
variation of results of the robustness test indicates that this purposed method is not significantly responsive to
small variations in analytical conditions. All parameters evaluated were in agreement with international
standardization, demonstrating that this UV method can be feasible applied to quantify manidipine in dissolution
and stability assays.
88
LC-MS DETERMINATION OF ANTIDEPRESSANTS IN LACTANTS MILK
Fernanda Rodrigues Salazar
Felipe D'Avila
Marcella H de Oliveria
Pamela Lukasewickz Ferreira
Ana Maria Bergold
Introduction: The use of medications during lactation is a common practice; however pharmacological
treatments impose serious doubts about the safety of drugs use during this time. Antidepressants can be
prescribed in this period and most of these drugs are excreted in breast milk. A validated bioanalytical method
using liquid chromatography coupled to mass spectrometry was applied to determine the excretion of five
antidepressants (fluoxetine, sertraline, citalopram, paroxetine and bupropion) in the human milk and estimate the
percentage of the ingested maternal drug that is being received by the infant through the breast milk. Materials
and Methods: Samples of human milk were obtained from mothers in lactation period and collected at two
hospitals - Santa Casa de Caridade de Bagé (Aproval 11-2012) and Hospital Materno Infantil Presidente Vargas
(Aproval 831302). LC conditions were validated in isocratic conditions with total run time of 15 min. A protein
precipitation cleaning procedure was applied for sample preparation using acetonitrile as precipitation agent. The
supernatant was removed and filtered directly to a vial and analyzed. Results and Discussion: A total of nine
samples were collected and analyzed. Concentrations found in the breast milk ranged from 11,8 to 156,5 ng/mL
for fluoxetine and 37,1 to 105,4 ng for sertraline. In all samples, values for relative dose higher than 6% were not
obtained. It is possible to say that the babies were getting only ~6% of the maternal drug dose. Values of relative
dose up to 10% can be considered of safe use during lactation period. Conclusions: Law concentration values of
the analyzed drugs were obtained in this study breast milk and for relative dose; so the use of fluoxetine and
sertraline
Acknowledgments
Capes/Brazil
Ethical approval
UFRGS, Hospital Materno Infantil Presidente Vargas, Santa Casa de Caridade de Bagé
89
DEVELOPMENT AND VALIDATION OF AN ULTRA-FAST LIQUID CHROMATOGRAPHY
METHOD FOR DETERMINATION OF ROSMARINIC ACID IN PORCINE SKIN
Flávia Nathiely Silveira Fachel
¹ Pharmaceutical Sciences Graduate Program, Federal University of Rio Grande do Sul, Porto Alegre, RS, Brazil.
Bruna Medeiros Neves
Introduction: Rosmarinic acid (RA) is a polyphenolic compound commonly found in several medicinal
plants. Since the presence of reactive species of oxygen has been suggested to be one of the main factors for
the skin aging, RA has been investigated due to its ability to eliminate these species. In this context, the present
study aim to develop and validate an Ultra-Fast Liquid Chromatography (UFLC) method for determination of RA in
porcine skin matrix, for further studies of new formulations exploring antioxidant activities and anti-aging process.
Methods: RA analyses were performed using an UFLC system device coupled to photodiode array detection.
Chromatographic separation was carried out in a C18 column (100 x 2.1 mm i.d.; 2.6 µm). The mobile phase,
which was composed of 17% acetonitrile and 83% water, acidified with 0.1% trifluoroacetic acid (v/v), was set at
isocratic mode, carried out at 55 ºC with a flow rate of 0.55 mL.min-1. The validation was performed according to
FDA guideline. Porcine ears were obtained from a local slaughterhouse. Results and discussion: Matrix solution
was extracted from full-thickness porcine ear skin with 2 mL of methanol in an ultrasonic bath for 45 min. The
UFLC method specificity was confirmed by injecting blank matrix and by the peak purity evaluation of RA in
presence of the matrix. The linearity in the range of 0.5-10.0 µg.mL-1 was proven by analysis of the calibration
curve of RA standards diluted in the mobile phase (r=0.999) and RA spiked with matrix (r=0.998). No matrix effect
(ME) was observed when the slopes of calibration curves were compared (ME%=3.61). The method was precise
and accurate, with respective relative standard deviation between 1.02-5.37% and recovery within the 85.21% to
88.63% range for the four levels analyzed. The methods robustness was confirmed using the Plackett-Burman
experimental design. Conclusions: The developed method was successfully validated for RA bioanalytical assay
in porcine skin.
Financial Support
Financial support from CNPq, FAPERGS and grants from CAPES.
90
DISSOLUTION TEST FOR DRONEDARONE IN TABLETS AND COMPARISON OF IN
VITRO DISSOLUTION PROFILES
Francieli Bellé
Department of Industrial Pharmacy, Federal University of Santa Maria, Santa Maria, RS, Brazil
Ana Isa Pedroso Marcolino
Postgraduate Program in Pharmaceutical Sciences, Federal University of Santa Maria, Santa Maria, RS, Brazil
Laís Engroff Scheeren
Priscila M. Bernardi
Postgraduate Program in Pharmaceutical Sciences, Federal University of Rio Grande do Sul, Porto Alegre, RS, Brazil
Joana Rodrigues Fernandes
Letícia Bueno Macedo
Daniele R. Nogueira-Librelotto
Department of Industrial Pharmacy and Postgraduate Program in Pharmaceutical Sciences, Federal University of Santa Maria, Santa
Maria, RS, Brazil
Clarice M. Bueno Rolim
Maria, RS, Brazil
Introduction: Dronedarone (DRO), an analogue of amiodarone, is a new antiarrhythmic agent. The
present study aims to develop a dissolution test for tablets containing 400 mg of DRO, as well as to apply
different comparison methods to the dissolution profiles obtained. Methods: The dissolution test was carried out in
a PharmaTest multi-bath (n = 6) dissolution system, in accordance with the United States Pharmacopeia under
the general chapters. The release characteristics of the film-coated tablets were determined using USP Apparatus
II (paddle) at 75 rpm in 900 ml of medium maintained at 37 ± 0.5°C. The dissolution media consisted of sodium
acetate buffer (pH 4.5) (Dissolution test I) and 1.0% (w/v) polysorbate 80 in water (Dissolution test II). Each
dissolution test was performed with 12 film-coated tablets, using manual sampling aliquots of 5 mL which were
withdrawn at time intervals of 15, 30, 45, 60, 75 and 90 min. The samples solutions were analyzed by a validated
UV-spectrophotometric method to characterize the dissolution profile of DRO. Results and Discussion:
Dissolution of DRO tablets was faster in pH 4.5 sodium acetate buffer with 89% of DRO released at 60 min. The
drug release profiles of Dissolution Test I and Dissolution Test II were compared by a model-independent
approach using the similarity factor (f2), which resulted in 56.47, suggesting that the two dissolution profiles were
similar. However, the dissolution profiles were also compared by calculation of dissolution efficiency (DE%) in 60,
75 and 90 min time points. One-way ANOVA of the DE% values showed that the percent of drug released was
not similar between the two media, being the sodium acetate buffer pH 4.5 the one with the higher DE% (65.83 ±
3.9 versus 58.59 ± 3.2 in 90 min). Conclusions: The dissolution test demonstrated to be adequate to evaluate the
drug release profile, and could be used in quality control of DRO film-coated tablets, collaborating to the official
codes.
Financial Support
CNPq (grant#401069/2014-1 and 447548/2014-0) and FAPERGS (grant#2293-2551/14-0)
Acknowledgments
The authors thank the Brazilian foundations CNPq, CAPES and FAPERGS for scholarships
91
EVALUATION OF INSULIN GLARGINE BY SIZE EXCLUSION LIQUID
CHROMATOGRAPHY METHOD
Gabriel Lunardi Remuzzi
Fernanda Pavani Stamm Maldaner
Leonardo Correa Cardoso
Vanessa Grigoletto Schramm
Ingrid Pereira da Silva
Introduction: Human insulin is synthesized from beta cells of the islets of Langerhans in the pancreas
and is the most important peptide hormone regulator of glucose homeostasis. The insulin glargine is a long-acting
recombinant human insulin analogue, produced by DNA technology in Escherichia coli, that contains 53 amino
acids in two polypeptide chains (A and B), linked by two disulfide bonds. It is currently being used worldwide for
the treatment of diabetic mellitus. The aim of this study was to validate parameters of a size-exclusion liquid
chromatography (SE-LC) method for the assessment of insulin glargine in biopharmaceutical formulations.
Methods: The SE-LC method was carried out on a BioSep-SEC-S 2000 column (300 mm X 7.8 mm i.d.)
maintained at 25 °C. The mobile phase consisted of 0.03 M MES acid, pH 2.5, run isocratically at a flow rate of
0.6 mL/min with PDA detection at 200 nm. Results and discussion: The chromatographic separation was
obtained with the retention time of 9.9 min and was linear over the concentration range of 0.02 - 180 µg/mL
(R2=0.9999). The accuracy was 99.38% with bias lower than 0.86%. The specificity was established in
degradation studies, which also showed that there was no interference of the excipients. The method was applied
for the analysis of biopharmaceutical formulations giving content of aggregates < 0.28%, and potencies between
96.25% and 102.67%. Conclusions: The SE-LC method was applied for the analysis of insulin glargine,
demonstrating the quality of the preparations in clinical use, contributing to improve the quality control, and to
assure the therapeutic efficacy of the biotechnology-derived product.
Financial Support
CAPES and FATEC.
92
DEVELOPMENT OF STABILITY-INDICATION METHOD BY LIQUID CHROMATOGRAPHIC
WITH FLUORESCENCE DETECTION TO DETERMINE GEMIFLOXACIN AND IMPURITIES
GABRIELA ROSSI BRABO
ANDRESSA TASSINARI DA SILVA
LISIANE BAJERSKI
MARTIN STEPPE
Postgraduate Program in Pharmaceutical Sciences, University Federal of Rio Grande do Sul - UFRGS, Porto Alegre, RS, Brazil
ELFRIDES EVA SCHERMAN SCHAPOVAL
Postgraduate Program in Pharmaceutical Sciences, University Federal of Rio Grande do Sul - UFRGS, Porto Alegre, RS, Brazil
Introduction. Gemifloxacin mesylate is a synthetic broad-spectrum antibacterial agent that provides
Gram-positive, Gram-negative and atypical pathogen coverage with a single agent taken daily. The drug is also
active against drug-resistant Streptococcus pneumoniae and multidrug-resistant Streptococcus pneumonia. The
literature researched describes the determination of the drug in biological fluids and pharmaceutical formulation,
but few studies aiming the impurities determination. Then, the main objective of this studies was the development
of an analytical method by high-performance liquid chromatography using fluorescence detection for quantitative
determination of the gemifloxacin - A, the main synthesis impurity - B and a degradation product formed on
alkaline hydrolysis conditions - C. Methods. The chromatographic separation was performed in a Supelco ® RP18 column (150 x 4.6 mm, 5 µm) at 30°C using a Shimadzu Prominence ® liquid chromatograph. The separation
was performed by means of a gradient elution (eluent A, acetonitrile 80%; eluent B, 0.3% triethylamine pH 3.0).
The gradient was a follows: 70% B (0 - 7 minutes), 70-40% B (7.01 - 10 minutes), 40% B (10.01 - 13 minutes), 4070% B (13.01 - 15 minutes) and 70% B (15.01 - 17 minutes) at a flow-rate of 1.2 mL min-1. The drugs were
evaluated using fluorescence detection at 344 nm as excitation wavelength and 399 nm as emission wavelength.
Results and Discussion. The results showed that the analytical method developed has excellent parameters of
system suitability. The capacity factor ranged between 5.6 and 12.9 and the asymmetry peaks in the range of 1.5
and 1.8. The lowest resolution between the peaks was 3.8 minutes and the theoretical plates were 3786, 5204
and 52643 for the drug A, B and C, respectively. Conclusion. The results confirm that the proposed method can
be used for routine analysis for quantitative determination and to verify the stability of this drug, but a complete
validation is necessary.
Financial Support
PDA UNIPAMPA
Acknowledgments
UNIPAMPA
93
Assessment of lisdexamfetamine dimesylate stability and identification of its
degradation product by NMR spectroscopy
Graciela carlos
Pharmacy Graduate Program, University of Rio Grande do Sul, Porto Alegre, RS
Alvicler Magalhães
UState University of Campinas, Chemistry Institute, Campinas, SP, Brazil
Ana Cristina Isler
State University of Campinas, Chemistry Institute, Campinas, SP, Brazil
Eloisa Comiran
Pedro Eduardo Fröehlich
Introduction: It is fundamental in drug therapy that pharmaceuticals products are safe and effective. A
key component of the quality and safety of pharmaceutical drugs is the control of impurities. Lisdexamfetamine
dimesylate (LDX), a long-acting prodrug stimulant indicated for the treatment of the attention-deficit/hyperactivity
disorder (ADHD), was subjected to forced degradation studies by acid and alkaline hydrolysis and the degradation
profile was studied. Methods: To obtain between 10-30% of degraded product, acid and alkaline conditions were
assessed with solutions of 0.01 M, 0.1 M, 0.5 M and 1 M of DCl and NaOD and these solutions were analyzed
through 1H NMR spectra. Results and Discussion: NMR spectra (1H NMR, 13C NMR, HSQC and HMBC) of
LDX solutions subjected to 1M (4+12 h) acid and alkaline hydrolysis were evaluated and one degradation product
(DP) was identified as a diastereoisomer of LDX. Acid and alkaline hydrolysis produced, respectively, 17.75% and
22.91% of the DP. Conclusions: The first structure DP of LDX was elucidated as a diastereoisomer of LDX
without their physical separation and it is very importante for LDX control of impurities.
Financial Support
CAPES
94
GREEN CHEMISTRY FOR THE DEVELOPMENT AND VALIDATION OF THE
MEASUREMENT SPIRONOLACTONE METHOD
Helder Lopes Vasconcelos
Universidade Estadual do Oeste do Paraná (UNIOESTE)
Ana Paula Pivotto
Isabela Angeli de Lima
Universidade Estadual do Centro Oeste (UNICENTRO)
Introduction: The Brazilian Pharmacopeia establishes the spectrophotometric method of molecular
absorption to quantify spironolactone using methanol as solvent. However, this compound is toxic to both handler
and environment, as well as flammable since it produces wastes that ask for specific and difficult treatment.
Thus, this trial aimed at replacing methanol by other solvents of lower toxicity and cost in order to minimize such
effects, and validate the method. Methods: Spironolactone solubility tests were carried out with various solvents
(diluted solutions of HCl, H2SO4, HNO3, NaOH, and Na2CO3; ethanol and acetone). Then, they were taken to
the spectrophotometer to determine the wavelength of maximum absorbance. To validate this alternative method,
the method was submitted to the analyses already established by RE 899/2003 to prove that its effectiveness in
quantifying such drug is equivalent to methanol effectiveness. Parameters such as linearity, limits of detection
(LOD) and quantification (LOQ), precision, accuracy and robustness were determined. Results and Discussion:
The results concerning solubility and scan spectrum showed the best answers when ethanol was used as
solvent, whose maximum absorption was at 238nm. The method was linear (r=0.996), with low LOD and LOQ
(0.0003 and 0.0009%, respectively) and high accuracy (RSD 1.3%). The accuracy was satisfactory, and its
recovery values varied from 91.2 to 95.5%. When there was some temperature variation (22 to 26°C), it was
observed that the obtained spironolactone recovery levels (102.4 and 103.3%, respectively) were satisfactory and
there was no significant difference between them. This can imply that the applied methodology was robust.
Conclusion: The method with ethanol as a solvent showed satisfactory results and it was under legislation for all
figures of merit. Consequently, it has been a viable, safer and cheaper alternative to analyze the target drug in
accordance with green chemistry principles.
Financial Support
FUNDAÇÃO ARAUCÁRIA
Acknowledgments
FUNDAÇÃO ARAUCÁRIA and UNIOESTE
95
SIMULTANEOUS QUANTIFICATION OF THREE PHLOROGLUCINOL IN THE EUGENIA
UMBELLIFLORA GREEN FRUITS EXTRACT BY LC-UV AND EXTRACTION
OPTIMIZATION
Ingrid Vicente Farias
Universidade do Vale do Itajaí (Univali)
Clarrissa M. AMORIM
Silvana S. R. NORBERTO
Valdir CECHINEL-FILHO
Tania M. B. BRESOLIN
Christiane MEYRE-SILVA
Eugenia umbeliflora belongs to the family Myrtaceae, popularly known as baguaçu. Eugenial A, B, C, D
and E had been previously isolated from green fruits, which presented antibacterial activity, being Eugenial C, D
and E the most active compounds. Therefore, this work aims to develop and validate a LC-UV method to the
simultaneous quantification of Eugenial C, D and E in hidroethanolic extract of green fruits of E. umbeliflora and
apply it in the extraction optimization process. It has applied the factorial design, with two variables, the ratio
drug:solvent (1:10, 1:15 and 1:20) and extraction time (4, 6 and 8 h); the extracts were prepared in triplicate
totaling 15 extracts in 5 levels. For the LC methodology it was used a Kinetex C18 column (150 x 4.6 mm x 2.6
µm) of Phenomenex®, at 40 °C. The mobile phase was a gradient of acetonitrile (0.125% formic acid), methanol
and acidified water at 0.7 mL/min, monitoring at 300 nm. The method was validated according to the parameters
of precision, linearity, limit of detection (LD) and limit of quantitation (LQ). The analytical methodology exhibited
good linearity with a correlation coefficient (r) higher than 0.99 and precision (RSD% < 5%). The LD and LQ for
Eugenial C were 0.55 and 1.68 µg/mL, for Eugenial D were 0.44 and 1.33 µg/mL and for Eugenial E were 0.28
and 0.84 µg/mL, respectively. The best conditions to extraction Eugenial C and Eugenial D was the level 3 (1:15;
6h) which showed 4.79 mg/g and 3.48 mg/g, respectively and to extracted the Eugenial E (7.01 mg/g) was level 4
(1:20, 4h). With these results it can be concluded that the method was reliable and observe that the ratio drug:
solvent and the extraction time influence the extraction of isolated E. umbelliflora compounds.
Financial Support
Programa de Bolsa de Iniciação Científica ¿ ProBIC/UNIVALI, CNPq.
Acknowledgments
Programa de Bolsa de Iniciação Científica ¿ ProBIC/UNIVALI, CNPq.
96
STABILITY-INDICATING LC METHOD FOR THE DETERMINATION OF DRONEDARONE
IN β AND HP βCYCLODEXTRIN INCLUSION COMPLEXES
Francieli Bellé
Daniele Rubert Nogueira-Librelotto
Maria, RS, Brazil
Clarice Madalena Bueno Rolim
Maria, RS, Brazil
Introduction: Dronedarone (DRO) is a new antiarrhythmic drug for the treatment of atrial fibrillation.
Cyclodextrins (CD) are cyclic oligosaccharides, which can form complexes with a variety of organic molecules.
This study aimed to develop and validate a simple stability-indicating liquid chromatographic (LC) method for
determination of dronedarone in inclusion complexes with βCD and HPβCD. Methods: The LC system was
equipped with a photodiode array detector. Separation and quantitation was made on a C18 analytical column
(250 × 4.6 mm i.d., 5 µm particle size). The detector was set at 289 nm. A mobile phase consisting of buffer
solution pH 4.9 (glacial acetic acid 0.3%) and acetonitrile (35:65, v/v) was used. The injection volume was 20 µL.
Forced degradation studies were carried out by subjecting standard and sample solutions to acidic, basic,
oxidative and photolytic conditions. The present method was validated following the ICH guidelines and
parameters such as specificity, linearity, precision, accuracy and robustness were evaluated. The inclusion
complexes were prepared by lyophilization, co-lyophilization and kneading followed by spray-drying. Results and
Discussion: Chromatographic separation was obtained within 6 min. Specificity of the method was evaluated by
verifying the purity of drug peak in stressed samples by PDA analysis, and no interference was found. The
analytical curve was linear in the 2.5-25 µg/mL concentration range. The precision was determined based on
relative standard deviation (RSD) and all repetitions presented values of RSD < 2.0%. Accuracy was assessed
and mean recovery was 99.77% (RSD = 1.38%; n = 9). Variation in system parameters did not have significant
effect in chromatographic pattern, indicating the robustness of the method. Conclusions: The validated LC method
was successfully applied to the quantitative analysis of DRO in inclusion complexes and can be employed for
drug analysis during stability studies.
Financial Support
Acknowledgments
The authors thank the Brazilian foundations CNPq, CAPES and FAPERGS for scholarships.
97
Analysis of organic impurities of besifloxacin by high-performance liquid
chromatography with gradient elution
Joanna Wittckind Manoel
Pharmaceutical Sciences Graduate Program, Federal University of Rio Grande do Sul
Julia Girardi
Faculty of Pharmacy, Federal University of Rio Grande do Sul
Martin Steppe
Nádia Maria Volpato
Introduction: Analysis of active and inactive ingredients is very important to ensure quality and safety of
ulk material and final products. Therefore, drug impurities research becomes an important requirement for assuring
total quality. Few studies were found regarding possible impurities in besifloxacin (B) raw material. In order to
evaluate these impurities, very sensitive analytical techniques should be employed due to low concentrations of
analytes. The HPLC is a method that contemplates the needs of this work. Methods: The analysis were
performed in a LC apparatus Shimadzu 20 A using a cyano column. For gradient mode operation, the mobile
phase was composed by 0.5% solution of triethylamine pH 3.0 and acetonitrile. The detection wavelength was
set at 330 nm and the column temperature was maintained at 25°C. Results and Discussion: The procedures
have been validated according to international guidelines, obtaining results within acceptable limits. The run time
was 13 min and the methods presented a linear response from 140 to 260 µg/mL for B and from 0.3 to 2.3 µg/mL
for the impurity A. With the injection volume of 20 µL, the limit of detection and quantitation were, respectively,
0.07 and 0.3 µg/mL. The RSD for inter-day precision was 6.36% for impurity A. Accuracy and robustness showed
satisfactory results. In raw material of B and pharmaceutical product of the drug used in this study, three related
impurities more polar than B were present but the impurity A was not detected. Conclusions: The proposed
method, considering the limit of quantitation, can be used in quantitative determination of impurity A in the
analysis of B raw material, as well as in ophthalmic suspension of the drug.
Acknowledgments
CNPq
98
VALIDATION OF A SIMPLE REVERSED PHASE-HPLC METHOD FOR DETERMINATION
OF BACLOFEN IN TABLETS
Juliana dos Santos
Priscila Rosa
Andréa Inês Horn Adams
Introduction: Baclofen is a muscle relaxant used as a first option in the spasticity treatment and muscle
spasms in patients with spinal cord injuries, being marketed as tablets of 10 mg. The official assays recommend
an ion pairing in situ method, which requires special reagents, turns the analysis longer and more expensive. The
objective of this study was to validate a simple reversed phase method for determination of baclofen in tablets.
Methods: The method employed a Shimadzu chromatographic system, equipped with PDA detector, mobile
phase composed by triethylamine solution 10 mM pH 7.0, methanol and acetonitrile (80:15:5), flow of 1.0 mL/min,
detection at 220 nm and RP-18 column. The linearity was studied in the concentration range of 2.5-100 µg/mL.
The accuracy was carried out by the spiking method and the precision was evaluated in two levels: repeatability
and intermediate precision. Sample solutions were exposed to stress conditions to study the method specificity.
The robustness was evaluated by a factorial design 23 and the chosen factors were the pH of aqueous phase,
proportion of methanol and flow rate of mobile phase. Results and discussion: The assay conditions resulted in a
baclofen retention time of around 6 minutes. The method was linear in the concentration range studied (r = 0.9999,
with linear regression and absence of deviation from linearity, α = 5%). The mean recovery was 99.3% (n= 9,
RSD = 0.8%), confirming the method accuracy. Likewise, the method is precise, since in both studied levels the
RSD values were < 2%. In all situations of stress test (acidic, basic, oxidative, thermal and UV radiation) the
baclofen peak was pure, demonstrating the method specificity. The robustness assessment showed no significant
effect of individual factors or of their interaction. Conclusions: It was developed a simple, fast and stability
indicative method, suitable for dosing baclofen in tablets. Financial support: Fipe Sênior/UFSM.
Financial Support
Fipe Sênior/UFSM
99
PHOTODEGRADATION KINETIC OF SILDENAFIL
Karine Favero Nicorena
Universidade Federal do Pampa, Uruguaiana, RS, Brasil.
Willian Ricardo da Rosa Almeida
Fabiana Ernestina Barcellos da silva
Stability studies are an important part of preformulation in dosage-form development and forced
degradation studies provide data to support identification of possible degradants or validation of stability indicating
analytical procedures. In this context, photostability studies provide information on effects of UV radiation on
pharmaceutical substances/products and support the choice of packaging material¹. The influence on the
photochemical decomposition of SLD was investigated in two different solvent, methanol and water. Aqueous and
methanolic solutions containing 500 µg mL-1 of SLD were transferred for quartz cuvettes and exposed to UV
Lamp 254 nm for two hours. Degradation rate kinetics was determined by plotting concentration of the remaining
drug versus time (zero-order process) and logarithm concentration of remaining drug versus time (first-order
process). Kinetic parameters such as apparent order degradation rate constant, half-life and t90 (time where 90%
of original concentration of the drug is left unchanged) were obtained from slopes of straight lines at each
condition. A Shimadzu LC equipped with an SPD-M20A PDA detector was used to analyze the degraded
samples on a Thermo Scientific C18 column (4.6 x 100 mm i.d) at 254 nm. A mixture of acetonitrile:triethylamine
60:40, v/v was used as the mobile phase at a flow rate of 1 mL/min. Plot of the logarithm of the percentage parent
compound remaining versus time was linear r > 0.992; k= 1.35 x10-4 s-1 using aqueous solvent, indicating that
the decomposition followed a first-order reaction. For methanolic solution, plot of the percentage parent compound
remaining versus time was linear r > 0.998; k= 4.83 x 10-3 µg mL-1s-1, indicating that the decomposition followed
and zero-order reaction. In conclusion, this approach demonstrated the influence of two solvents with different
polarities at photodecomposition of SLD and can be guidance in studies to define phosphodiesterase 5 inhibitors
degradation pathways.
100
DEVELOPMENT AND VALIDATION OF ANALYTICAL METHOD FOR 4PHENYLTELLURO-7-CHLOROQUINOLINE IN PHARMACEUTICAL DOSAGE FORM
Kelly Ayumi Nakama
Federal University of Pampa
Carlos Eduardo Rosa da Silva
Willian Goulart Salgueiro
Bruna Goldani
Federal University of Pelotas
Diogo Alves
Federal University of Pelotas
Daiana da Silva Avila
Sandra Elisa Haas
Introduction: Many pathological conditions remain without an effective treatment and cure, but the ones
whose molecular pathogenesis is linked to redox imbalance, are especially prevalent and dangerous. Thus, the
study of new molecules is an important step towards effective novel therapies. Among many candidates to act as
a free radical neutralizer, organochalcogens (OG) have been described for having a great spectrum of action over
different disease models. 4-phenyltelluro-7-chloroquinoline (TQ), a novel OG that is capable to exert antioxidant
action not only in vitro, but also in vivo by modulating key molecular pathways in Caenorhabditis elegans. The
aim of this work is to develop an HPLC - PDA method able to quantify TQ in solution. Methods: Stock solutions
of standards were prepared by dissolving TQ in acetonitrile. HPLC-PDA (Shimadzu) was used with a C18-RP
column, triethylamine water/acetonitrile as mobile phase. The validation procedures were performed according to
ICH guidelines and RE nº899/2003-ANVISA. Results and Discussion: Method was validated according to
Linearity (r>0.999), precision with maximum RSD of 1.54% and accuracy was 103.60%. Specificity showed not
interference and peak purity was 1.00. Robustness were evaluated with small variations in mobile phase, pH and
flow rate the result obtained was 100.64% for all conditions. No interference was observed in the chromatograms
during analysis. Conclusion: The method was developed and validated for TQ quantification in pharmaceutical
dosage forms.
101
VALIDATION OF LC METHOD TO ASSAY ROSMARINIC ACID/ CYCLODEXTRIN
COMPLEXES
Kleyton Santos Veras
Pharmaceutical Sciences Graduate Program, Federal University of Rio Grande do Sul, Porto Alegre, RS, Brazil
Flávia Nathiely Silveira Fachel
Valquíria Linck Bassani
Amélia Teresinha Henriques
Introduction: Rosmarinic acid is a hydroxycinnamic phenolic compound, presenting antioxidant potential
as its remarkable activity. The oral route is the main pathway of drugs/medicines administration since it is
comfortable, easy and cheap. Nevertheless, rosmarinic acid has a low aqueous solubility, a feature that
decreases its dissolution in gastrointestinal fluids, consequently affecting its bioavailability. A technological
approach to overcome this physicochemical property is its complexation with cyclodextrins (CD). Thus, the aim
of the work is develop and validate a liquid chromatography (LC) method to assay rosmarinic acid complexes for
further applications. Methods: Selectivity (βCD, HPβCD and MβCD), linearity, precision, accuracy, detection and
quantification limits were assessed according to ICH guidelines. The analyses were performed on a reversedphase C18 column (150 x 4.6 mm; 5µm) at 30 °C and under UV detection at 330 nm. The mobile phase was
composed of acetonitrile: trifluoroacetic acid 0.1% (25%) and water: trifluoroacetic acid 0.1% (75%) (pH 2.3) and
was eluted at isocratic flow rate of 1.0 mL.min-1. Results: Rosmarinic acid presented retention time of 6.1
minutes, theoretical plates over 5,000 and tailor factor under 1.1. The method showed to be selective and linear in
the concentration range 0.5 - 10 µg/mL with R-squared higher than 0.999. The intra-day and inter-day precision
have demonstrated R.S.D lower than 5%, accuracy between 90 to 110% and low detection (0.04 µg/mL) and
quantification (0.13 µg/mL) limits. Conclusion: The method was simple, quick and able to quantify rosmarinic
acid/CD complexes with precision and accuracy.
Financial Support
FAPERGS
102
FOURIER TRANSFORM INFRARED SPECTROSCOPY ANALYSIS AND
PHOTOSTABILITY STUDIES OF DOXORUBICIN-LOADED CHITOSAN NANOPARTICLES
LAIS ENGROFF SCHEEREN
Universidade de Barcelona
Introduction: Nanoparticles (NPs) have demonstrated greater potential to trigger the encapsulate drug
according to pH. Thus, the pH-sensitive lysine-based surfactant 77KS has been studied as an adjuvant into NP
matrix to promote doxorubicin (DOX) release at acidic tumor tissue and intracellular compartments. Furthermore,
adjuvants like PEG and poloxamers have been used to extend the residence time of NPs in blood stream and
overcome the multidrug resistance phenomena, respectively. This work aims to evidence the incorporation of
DOX, 77KS, and the copolymers PEG and poloxamer 188, into the pH-responsive chitosan nanoparticles (CSNPs) previously developed by our group. Moreover, to verify the ability of the NP suspensions and lyophilized
samples to protect DOX from UVA radiation. Methods: CS-NPs were prepared by an ionotropic gelation method,
using 77KS, PEG and poloxamer as adjuvants into the NP matrix. Each NP suspension was submitted to
lyophilization process to obtain the dry formulations, which were then analyzed by Fourier Transform Infrared
Spectroscopy (FT-IR). Moreover, the lyophilized samples and NP suspensions were compared concerning their
photostability after exposition to UVA radiation during 48h. Results and Discussion: DOX-CS-NPs were optimized
with a final CS:TPP:77KS:DOX ratio equal to 5:2:0.5:0.5 (w/w/w/w). FT-IR analysis showed the cross-linking
between CS and TPP, as well as the load of DOX into the NPs. Likewise, the incorporation of 77KS, PEG and
poloxamer was also evidenced. Finally, liquid chromatography analysis of the samples from stability studies
showed that the dry formulations were able to protect DOX from photodegradation, with t1/2 7.5-fold greater than
for NP suspensions. Conclusions: The overall findings demonstrated that the method to prepare the NPs
including different adjuvants was efficient, as proved by FT-IR analysis. Moreover, the lyophilized NPs were able
to improve DOX photostability, suggesting a higher storage time.
Financial Support
CNPq and FAPERGS
Acknowledgments
CNPq and FAPERGS
103
VALIDATION OF A RP-LC METHOD AS A SUITABLE APPROACH FOR THE
CHARACTERIZATION OF METHOTREXATE-LOADED CHITOSAN NANOPARTICLES
Maria, RS, Brazil
Francieli Bellé
Maria, RS, Brazil
Introduction: Methotrexate (MTX) is well established for the treatment of solid tumors and leukemias. In
order to improve the delivery of this drug in its specific site of action, we designed pH-responsive MTX-loaded
chitosan-based nanoparticles (CS-NPs). Moreover, here we developed and validated a reversed-phase liquid
chromatography (RP-LC) method to quantify the encapsulated drug. Methods: CS-NPs were spontaneously
obtained by ionotropic gelation technique, using tripolyphosphate (TPP) as a cross-linker agent. The anionic
surfactant derived from lysine (77KS, with sodium counterion) was used as a pH-sensitive adjuvant. NPs were
characterized using a Malvern Zetasizer equipment. The RP-LC method was carried out on a C18 column, with
mobile phase consisted of 0.05 M potassium phosphate buffer pH 3.2 and acetonitrile (86:14, v/v), with UV
detection set at 303 nm. The method was validated with evaluation of the parameters specificity, linearity,
precision, accuracy and robustness. Results and Discussion: The pH-responsive NPs were optimized with
CS:TPP:77KS ratio of 5:1:0.5 (w/w/w). They showed mean size in the range of 162-183 nm, and positive zeta
potential values (> 22 mV). The proposed analytical methodology was validated and provided results within the
specifications recommended by official guidelines. The specificity tests showed that there was no interference
from the polymer or surfactant. The RP-LC method was successfully applied for the analyses of MTX in different
batches of NPs. Moreover, it was evidenced as a suitable approach to determine the drug entrapment efficiency,
as well as to quantify MTX during in vitro release assays and photostability studies. Conclusions: Altogether, the
results proved that the validated method is useful for the quantitative analysis of MTX in complex matrices of
nanotechnology-based polymeric formulations. Moreover, it can be inferred that these NPs have potential
application as new vectors for cancer therapy.
Financial Support
Acknowledgments
The authors thank the Brazilian foundations CNPq and CAPES for scholarships.
104
Development and validation of a novel stability-indicating HPLC method for the
quantitative determination of fucoxanthin, a marine carotenoid
Leticia Malgarim Cordenonsi
Pharmaceutical Sciences Graduate Program, College of Pharmacy, Federal University of Rio Grande do Sul
Andressa Santer
Rafaela Martins Sponchiado
Martin Stepp
Renata Platcheck Raffin
Postgraduate Program in Nanoscience, Centro Universitário Franciscano
Elfrides Eva Scherman Schapoval
Introduction: Recently, use of natural products available from marine sources are receiving more
attention. Fucoxanthin (FUCO) is a marine carotenoid exhibiting several health benefits and the anti-cancer effect
is well documented. The aim of this work was to establish a simple and rapid stability-indicating high performance
liquid chromatograph (HPLC) method to evaluate simultaneously FUCO in power and oil. Methods: The validation
of the method was accomplished of agreement the parameters of the main Official Guides. The analyses were
performed using HPLC system equipped with PDA detector. During the development of the method, variations on
analytical conditions such as pH, proportion of solvents and different temperature columns had been tested
seeking the best chromatographic conditions. The evaluation of the system suitability was done through the
observation of analytical parameters such as retention time, tailing factor and efficiency. Results and Discussion:
A successful resolution and retention times were obtained with on a C18 column kept at 40 °C, at a 1 mL min-1
flow rate, wavelength of 450 nm and the injection volume was 50 µl. The mobile phase consisting a isocratic
system of acetonitrile:water (pH 7.0) (95:5 v/v). The method was found to be linear in the 1.0- 40.0 µg/ml
concentration range (r> 0.99) (α=0.05) and the limits of quantification were 0.1 μg mL-1. Repeatability was
determinate at three concentration levels obtaining a RSD always lower than 0.7% and the accuracy of the
method was greater than 97%. For robustness was evaluated using a factorial Box-Behnken experiment design,
producing results with showed a good correlation, there were no significant interaction effects and no evidence of
inadequacy were detected by the lack-of-fit test. Conclusion: The proposed conditions were successfully applied
for the analysis of FUCO in power and oil.
Financial Support
Capes
Acknowledgments
Pharmaceutical Sciences Graduate Program
105
ASSESSMENT OF STREPTOKINASE BY REVERSED-PHASE LIQUID
CHROMATOGRAPHY METHOD
Luis Gustavo Jung Motta
Rafaela Ferreira Perobelli
Clóvis Dervil Appratto Cardoso Júnior
Bruna Xavier
Sérgio Luiz Dalmora
Introduction: Streptokinase (STK) consists of a 414 amino acids polypeptide chain that does not contain
disulfide bonds, with a molecular mass of 47 kDa. It is clinically used as a thrombolytic agent to treat patients
with acute myocardial infarction, deep vein thrombosis, arterial thrombosis and embolism. The chromogenic
substrate assay (SCSA) has been recommended as the bioassay for the potency evaluation of STK in
biopharmaceutical formulations. The aim of this study was to develop a reversed-phase liquid chromatography
(RP-LC) method for the evaluation of STK correlating the results with the SCSA. Methods: A RP-LC method was
carried out on a Jupiter C4 column (250 mm x 4.6 mm i.d., 5 µm), Phenomenex, maintained at 25 °C. The mobile
phase consisted of 50 mM sodium sulphate buffer pH 7.0 and methanol (90:10, v/v), run isocratically at a flow
rate of 0.8 mL/min. with photodiode array (PDA) detection at 220 nm. The SCSA was applied for the evaluation of
the biological potencies of STK in biopharmaceutical products, calculated against the 3rd international standard of
streptokinase. Results and discussion: The chromatographic separation was obtained with the retention time of
19.3 min and was linear over the concentration range of 85 - 25 000 IU/mL (r = 0.9999). The limits of detection
and quantitation were 26 and 85 IU/mL, respectively. The accuracy was 100.24% with bias lower than 0.99%.
Moreover, the RP-LC method demonstrated acceptable results for precision and robustness. Biopharmaceutical
products of STK were analysed by the RP-LC and by SCSA, giving potencies between 95.32% and 101.75% of
the stated potency, with significant correlation (p>0.05). Conclusions: The method developed was applied for the
analysis of streptokinase in biopharmaceutical formulations, demonstrating the quality of the preparations, and
contributing to establish alternative which improve the quality control assuring the therapeutic efficacy of the
biotechnology-derived product.
Financial Support
CAPES and FATEC
106
ANALYSIS OF PHOTODEGRADATION KINETICS OF FINASTERIDE IN
PHARMACEUTICAL PREPARATIONS
Marcelo Donadel Malesuik
Rafael Henrique Dias Reis
Introduction: Finasteride is a synthetic drug widely used for the treatment of androgenetic alopecia,
benign prostatic hyperplasia, and prostate cancer. It´s an antiandrogen which acts by inhibiting 5αreductase, the
enzyme that converts testosterone to dihydrotestosterone. A daily dose of 5 mg has been used for the treatment
of benign prostatic hyperplasia and prostate cancer, and a 1 mg dose has been used for the treatment of
androgenetic alopecia. Stability of these drugs is a critical point of quality control and pharmacology of therapeutic
agents, since a non-stable or reactive molecule may influence their structure, efficacy and safety. Stability tests,
known as forced degradation studies, are the important tools used to detect, quantify and identify the generation
of degradation products. The photodegradation of finasteride was studied in order to investigate the degradation
kinetics of this drug. Methods: The degradation was carried out in acetonitrile, with raw material and tablets, in
quartz cells under UVC light at 254 nm. The analyses of the degraded samples were performed by validated
stability-indicating liquid chromatographic method. Results and Discussion: The photodegradation of finasteride
for both samples in acetonitrile solution shows a zero-order kinetics under the applied experimental conditions.
The kinetics parameters such as rate constants, half-life (t1/2) and the time when 90% of the drug original
concentration is left were determined. The obtained results confirm the reliability of the chromatographic method
for determining the kinetics run of finasteride in the presence of its degradation products. Conclusions: The
present study reveals the photolability of the drug in the tested conditions. Therefore, appropriated photoprotection
is recommended during the manipulation of the drug.
Financial Support
PDA-UNIPAMPA
107
DEVELOPMENT AND VALIDATION OF A HPLC METHOD TO ASSAY KAEMPFEROL IN
DIFFERENT MATRICES
Mariana Colombo
Gabriela de Lima Melchiades
Introduction: Kaempferol (KPF) is a flavonoid with different pharmacological properties, including
antineoplastic activity. Despite the cytotoxic capability, KPF has reduced water solubility and the incorporation
into a nanoemulsion seems to be a good strategy to improve its activities. Thus, the aim of this study was to
develop and validate a high-pressure liquid chromatography (HPLC) analytical and bioanalytical method to assay
KPF in different matrices for further application. Methods: The method was validated according to ICH and FDA
guidelines. Selectivity, linearity, limit of detection (LOD) and limit of quantification (LOQ), precision and accuracy
were evaluated in different matrices: KPF standard solution, KPF loaded-nanoemulsion, phosphate buffered saline
(pH 6.4) containing 2% polysorbate 80 (receptor fluid for in vitro release and ex vivo diffusion experiments) and
porcine nasal mucosa extract. The analyses were performed on a reversed-phase C18 column at 35 °C and under
UV detection at 368 nm. The mobile phase was composed of methanol:formic acid 0.1% (75:25, v/v) and was
eluted at an isocratic flow rate of 1.0 mL.min-1. Results and Discussion: The method was selective with no
matrix-related interference in the KPF retention time. The analysis was linear (0.25-7.5 μg.mL1) with a coefficient
of determination higher than 0.99; precise (relative standard deviation values for intra- and inter-day analysis were
0.10-1.97% and 0.28-3.37%, respectively and accurate (98.40-109.18%). The values of LOD (0.03-0.07) and LOQ
(0.10-0.22) were calculated using the respective calibration equations for each matrix. There was low matrix effect
during analysis (lower than 5%) and the concentration of KPF in all matrices remained constant after 48h of
storage at room temperature. Conclusions: The developed HPLC method is simple, quick and can be used to
determine KPF even at low concentrations in all investigated matrices.
Financial Support
This work was supported by CNPq.
108
DEVELOPMENT AND VALIDATION OF AN ANALYTICAL METHOD FOR ATROPINE
QUANTIFICATION IN LEAVES OF Atropa belladonna (L.) BY HIGH-PERFORMANCE
THIN-LAYER CHROMATOGRAPHY (HPTLC)
Mariana Koetz
Introduction: Atropine is an alkaloid known for acetylcholine inhibition and as antimuscarinic and is
present in leaves of Atropa belladonna (L). This plant is mainly employed in homeopathic preparations. The
importance of medicinal plants in pharmaceutical industry has driven scientific studies to develop analytical
methods for quantification of chemical markers, present in herbal drugs, and that meet the quality control
parameters of current legislation. A simple method of HPTLC has been developed and validated to quantify
atropine in belladonna leaves. Methods: Method was performed using silica gel 60 F254 HPTLC plates, toluene:
ethyl acetate: diethylamine (55:35:10) as mobile phase and atropine detection by densitometry in wavelength of
217 nm. The validation was performed by evaluating the parameters of selectivity and specificity, linearity, limits
of detection and quantification (LOD, LOQ), precision, accuracy and robustness. Results and Discussion: In our
analysis we obtained atropine spots with Rf = 0.36. Selectivity has been proven by adding a reference substance
in the sample solution, which produced an increase only peak area of the sample, without changing the area of
the adjacent peaks. Specificity was verified by comparing the UV profiles of reference and sample solutions.
Linearity was determined of reference solutions with concentrations range 320-814 µg/mL, with r2 = 0.9978. LOD
and LOQ were 43.05 and 143.5 µg/ml, respectively. Repeatability and reproducibility were RSD of 4.2 and 3.2%,
respectively. The average recovery level for accuracy was 97.3%. Robustness was evidenced by changes in
composition of mobile phase, plate scanning time, wavelength, and type of plate and all results showed a RSD
lower than 5%. The average content of atropine quantified in the material analyzed was 0.29%. Conclusions: The
developed method is effective for quantification of atropine in beladona leaves and meets the validation
requirements of current legislation.
Acknowledgments
I thank Professor Amelia T. Henriques, the graduate fellow pharmacognosy laboratory and the Faculty of Pharmacy at
UFRGS.
109
QUALITY CONTROL OF TIPRANAVIR CAPSULES BY CAPILLARY ELECTROPHORESIS
Matheus Wagner Lago
Pharmaceutical Sciences Graduate Program, Federal University of Santa Maria, Santa Maria, RS, Brazil
Mariane Lago Friedrich
Ana Paula Christ
Introduction: Tipranavir (TPV) is a drug of the protease inhibitors class, used in the treatment of AIDS. It
is considered the first option when HIV presents resistance to other protease inhibitors. There are no
pharmacopeial monographs of this drug, which justify the development of analytical methods to assay TPV
capsules. Methods: An Agilent 7100 equipment was used, with the following conditions: background electrolyte
composed by a mixture (95:5) of solution A and B (A, sodium borate, 50 mM pH 9.0, and B, methanol), a fused
silica capillary of 40 cm x 50 µm (effective length x inner diameter), kept at 25°C, voltage of 28 kV and detection
at 282 nm. The method validation was carried out according the ICH Guidelines. Furosemide was used as internal
standard. Results and discussion: The migration time of TPV was around 3 minutes in the conditions used. The
specificity was evaluated by stress test, and the residual content were 92.0% (HCl 0.1 M/10 days), 91.3%
(NaOH 0.1 M/10 days) and 89.3% (H2O2 3%/8 hours). The TPV peak was well resolved of the other peaks
observed and purity was observed in all conditions. The method was linear (r=0.999) in the range of 20 to 200
µg/mL with no significant linearity deviation and significant linear regression (α = 5%). Precision was evaluated by
intraday (RSD values of 1.1% and 1.7%) and inter-day assays (RSD=1.7%, n=12). The mean recovery (100.3%,
RSD=1.7%, n=9) indicated the accuracy of the method. The robustness was evaluated through factorial test 23,
and no influence of single factor or combination of them was observed in the method response. Conclusion: A
fast, environmentally friendly and stability indicating method to assay TPV capsules was developed, contributing
to improve the quality and safety of this pharmaceutical form. Financial Support: Capes. Acknowledgements:
Cristalia Brazil for providing tipranavir standard.
Financial Support
CAPES
Acknowledgments
Cristalia Brazil for providing tipranavir standard
110
Development and characterization curcumin nanoparticles Physical Chemistry
incorporated in a system semisolid
MURIEL PANDO PEREIRA
UNIVERSIDADE FEDERAL DO PAMPA
LISIANE BAJERSKI
FELIPE FERRARINI MACHADO
JOSÉ VALDIR DA SILVA CORRÊA JÚNIOR
SANDRA ELISA HAAS
Curcumin, a natural compound extracted of the saffron's rhizome (Curcuma longa L.), is a popular spice
of the Asian cuisine and has been studied due its wide range of pharmacologicals properties, altough it presents
poor stability and bioavailability. Nanotechnology is the area of pharmaceutical science involved in the
development, characterization and application of therapeutic systems at the nanometric or micrometric scale, as
an alternative to improve the physicochemical and pharmacokinetic characteristics of drugs. Delivery systems,
such as polymeric nanocapsules, can promote the release of drugs in a modified form via routes other than oral
and topical route. The aim of this study was to develop polymeric nanocapsules containing curcumin in order to
improve its therapeutic efficacy and stability, with subsequent incorporation into a semissolid system as an
pharmaceutical form. The polymeric nanocapsules were prepared by the dispersion of preformed polymers
method, obtaining the blank formulation (FB) and the formulation containing curcumin (HR). After that, pH, particle
diameter, dosing, encapsulating rate and zeta potential were evaluated. The formulations were incorporated in 3%
hydroxyethylcellulose gel to obtain the blank gel formulation (GFB) and the gel formulation containing curcumin
(GFC). Blank gel (GB) and free curcumin gel (CG) were prepared too. It was evaluated pH, particle diameter,
dosing and rheology. The polymeric nanocapsules containing curcumin had pH around 7.78, average diameter
around 0.255 µm, 122.17% dosing, encapsulating rate of 90.92% and zeta potential of -13.73 mV. The gels
containing curcumin formulation had pH around 6.13 , average diameter around 190.37 µm, dosing of 29.64 % and
pseudoplastic and non-newtonian behavior. The encapsulation of curcumin in polymeric nanocapsules proved to
be feasible, and its incorporation into semisolid systems is an innovative and promising form of modified release
of drugs for the topic route.
111
Development and validation of an analytical method for dosing azithromycin in
microemulsion systems
Mysrayn Yargo de Freitas Araújo Reis
State University of Paraiba
Maria Salete da Silva Rodrigues
Joyce Cordeiro Borges
Amaro César Assis de Lima
Larissa Pereira Alves
Joandra Maisa da Silva Leite
Ana Cláudia Gonçalves Santos
Airlla Laana de Medeiros Cavalcanti
Geovani Pereira Guimarães
Bolívar Ponciano Goulart de Lima Damasceno
Introduction: Azithromycin (AZ) is a semi-synthetic macrolide antibiotic that belongs to the azalides
class. It has a broad spectrum of action against Gram-negative bacteria. Brazilian and American Pharmacopoeias
recommend dosing of AZ by microbiological testing and high-performance liquid chromatography, respectively.
Microemulsions (MEs) are drug release vehicles widely used for solubilization of drugs. MEs can increase the
therapeutic efficacy of drugs and minimize side effects. This study aimed to validate a spectrophotometric
method UV-Vis that proved faster, lower cost and trustworthy for dosing of AZ in EMs. Methods: The method was
validated according to Resolution 899/2003 of the Agência Nacional de Vigilância Sanitária (ANVISA). The
parameters of selectivity, linearity and range, precision, accuracy and robustness were evaluated. The two ME
formulations (ME1= 40% aqueous phase; 55% surfactants, 5% oil phase/ME2= 30% aqueous phase; 65%
surfactants; 5% oil phase, both with 30 μg.mL1 of AZ) were diluted in triplicate using as solvent a mixture of
phosphate buffer and H2SO4 (1: 9) for the dosing of the AZ. The dilutions were incubated in a thermostatic bath
at 50ºC for 30 minutes. For the encapsulation efficiency 1 g of ME was centrifuged at 7000 rpm for 1 hour. After
centrifugation, aliquots of the supernatant were taken and diluted in the same conditions proposed before. Results
and discussion: The analytical method showed to be selective, with linearity in the range of 10-50 μg.mL1 with
correlation coefficient (r) 0.9936, detection and quantification limits of 1.85 and 6.17 µg ml-1, respectively. The ME
formulations showed encapsulation efficiency 23,99 e 29,91 µg.mL-1, for to ME1 and ME2 respectively.
Conclusion: The method was rapid, selective, linear, accurate and precise. Therefore the method can be used for
the dosing of the AZ in microemulsion systems.
Financial Support
CAPES, UEPB
112
IN VITRO DISSOLUTION METHOD FOR RIVAROXABAN TABLETS BASED ON IN VIVO
ABSORPTION PROFILE
Nathalie Ribeiro Wingert
Graduation Program in Pharmaceutical Sciences (PPGCF), UFRGS, Porto Alegre, RS, Brazil
Natália Olegario dos Santos
Sarah S. Campanharo
Nadia Maria Volpato
Martin Steppe
Introduction: Oral drugs with high permeability but poorly soluble in water, such as the anticoagulant
rivaroxaban (RIV) in instant-release tablets, have a major potential to reach high level in vitro and in vivo (IVIV)
correlation. Currently, there is no information on scientific literature regarding the evaluation of RIV dissolution
based on in vivo properties. Objective(s): This study aimed the development and validation of an in vitro
dissolution method based on in silico-in vivo data to determine whether an IVIV relationship could be established
for RIV tablets. Materials and Methods: RIV dissolution was monitored by HPLC along several sampling points.
Through method optimization, different conditions were evaluated in order to obtain a continuous and controlled
dissolution of RIV IR tablets. In silico modelling was applied to plasmatic concentrations and deconvolution
technique employed to obtain RIV fraction absorbed. The level of relationship between fractions absorbed (FA)
and dissolved (FD) was evaluated by linear regression. Results and Discussion: Plasmatic concentration (Cp) in
vivo values were modelled using computational simulation with adjustment of pharmacokinetic properties. Cp was
converted into FA by Wagner-Nelson deconvolution approach. Gradual and controlled dissolution of RIV tablets
was obtained with 30 rpm basket on 50 mM sodium acetate + 0.2 % SDS, pH 6.5 media. Dissolution was
conducted for up to 180 min. FA was plotted against fraction dissolved, and a linear point-to-point regression
(R2=0.9961) was obtained. Conclusions: A point-to-point linear in vitro-in vivo relationship was stablished for RIV
20 mg tablets based on modelled in silico values, equation that correlates FA and FD is: FA = 1.005FD + 0.0994.
Developed in vitro dissolution method was successfully validated and can be applied as a prognostic tool for drug
absorption performance and represents valuable data for quality control and drug development.
Financial Support
CAPES/Brazil
113
OPTIMIZATION OF CONDITIONS FOR THE BIOTRANSFORMATION OF
AMBRISENTANA BY CUNNINGHAMELLA ELEGANS
Rafaela Martins Sponchiado
Julia Medeiros Sorrentino
Leticia Malgarim Cordenonsi
Natália Olegário
Nadia Maria Voltato
Martin Steppe
Alexandre Meneghello Fuentefria
Introduction: Biotransformation is the chemical modification(s) made by an organism or enzymes on a
chemical compound and has been applied in metabolism studies, structural modification and synthesis of active
compounds. Drug biotransformation using fungi are considered an alternative approach to the asymmetric
synthesis and conventional in vivo and in vitro metabolism studies. Fungi of the genus Cunninghamella stand out
amongst other genera because can catalyze reactions mimicking the mammalian metabolism. The aim of this
study is the optimization of conditions for the biotransformation of ambrisentan using filamentous fungi. Methods:
Biotransformation study has been optimized with fungi Cunninghamella elegans ATCC 9245 in potato broth a
temperature 27 ºC. In this work, the parameters under evaluation are: influence of pH, potato broth concentration
and shaken condition. Every 24 h, a 3 mL aliquot of the liquid culture medium containing the drug and positive
(only fungus) and negative (only drug) control are aseptically collected and immediately extracted with 8 ml
dichloromethane and analyzed by HPLC. Results and discussion: Biotransformation medium was tested at two
different conditions: 100% and 50%. No difference was observed between concentrations, thus, the lower was
adopted. Another condition evaluated was drug stability with broth pH modification, since this drug is susceptible
in acid medium. The pHs 4.0 to 9.0 were verified and the pH 9.0 demonstrated the greatest result. However, at
this value, the fungus does not have the same metabolomics ability; therefore, pH 6 was chosen to ensure the
skill of fungi metabolizing and avoid significant degradation signal. Shaken tests are being conducted.
Conclusions: In this optimization study, important parameters were evaluated and demonstrated an improvement
in the formation of ambrisentan metabolites.
Financial Support
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
114
DEVELOPMENT AND VALIDATION OF A STABILITY INDICATING HPLC METHOD FOR
DETERMINATION OF DAPAGLIFLOZIN IN TABLETS
Rafaela Zielinski Cavalheiro de Meira
Universidade Estadual do Centro-Oeste
Larissa Sakis Bernardi
Paulo Renato de Oliveira
MEIRA, RZC*, BERNARDI, LS and OLIVEIRA, PR Post-Graduation program in Pharmaceutical
Sciences, Department of Pharmacy, UNICENTRO, Guarapuava-PR, Brazil. Introduction Dapagliflozin (DAPA) is
a highly potent, selective and reversible inhibitor of sodium-glucose cotransporter 2 (SGLT2). Its commercial
formula, Farxiga®, was approved by the FDA in January 2014 and by ANVISA in July 2013. So far, there is no
published stability indicating HPLC method for DAPA. Methods The chromatographic conditions were as follows:
Synergi Fusion C18 column (150 x 4.6 mm) maintained at 30 °C, injection volume of 20 µL, flow rate of 1.0
mL/min, running time of 10 min, mobile phase composed of acetonitrile and water acidified with 0.1% formic acid
(42:58, v/v) and detection at 245 nm. Reference sample solution was submitted to forced degradation in acid
media, basic, oxidative, heat, and photolytic. Results and Discussion Degradation products and significant
reduction of active area could be observed after exposure to UV light and daylight. The excipients did not show
any interference in the analysis. The developed method was linear in the range of 1 to 100 µg/mL (r2=0.9997). In
repeatability and between-analysts precision studies, DAPA showed RSD values lower than 5%. The method
demonstrated to be accurate within the range of interest, showing a mean recovery of 81.66, 100.47 and 119.35%.
The limit of detection and quantitation were 0.09 and 0.28 µg/mL, respectively. The method was robust across the
randomized variations (p>0.05). The analysis of tablets containing 5 and 10 mg of DAPA resulted in an assay of
99.47 and 101.33%, respectively. Conclusions Stress studies revealed that drug was most susceptible to
photolytic degradation, revealing new degradation products, whose analysis is underway to their identification.
The methodology was validated, and was successfully applied for the quantitative analysis of dapagliflozin in
pharmaceutical formulations.
115
DEVELOPMENT AND VALIDATION OF STABILITY INDICATING METHOD BY LC TO
DETERMINE LINAGLIPTIN IN THE PRESENCE OF POTENTIAL IMPURITIES
Raquel Balestri Heleno Ferreira
University Federal of Pampa (UNIPAMPA)
Flávio Dias Ferreira
Introduction. Linagliptin is an inhibitor of DPP-4 used for glycemic control of type 2 mellitus diabetes.
Since the drugs used in the production process of the pharmaceutical formulations are not considered totally pure,
the Regulatory Guides describing the need of the drug quantification and impurities potentials. Therefore, the goal
of this study was to develop a method by high-performance liquid chromatography to quantify the drug in the
presence of potential impurities. Methods. The chromatographic separation was performed in a Thermo
Scientific® RP-8 column (150 x 4.6 mm, 5 µm), at 30°C, using a Dionex Ultimate 3000® liquid chromatograph.
The separation was performed by means of a linear gradient elution (eluent A: formic acid 0.1% pH 3.5; eluent B:
acetonitrile). The gradient obeys the following sequence: 30-70% of B (0 - 4 minutes) and 70-30 of B (4,01 - 8
minutes) at a flow-rate of 0.6 mL min-1 and run time of 10 minutes. The injection volume was 20 μL and detection
at 293 nm using photodiode array. Results and Discussion. The method showed selectivity, since there was no
interference of degradation products formed in stress conditions (UV-A, basic and acid hydrolysis, base and acid
hydrolysis at 80 °C, and oxidation), excipients and three synthesis impurities. The response was linear over a
range from 0.87 to 144.0 μg mL-1 (r = 0.9997). The method showed suitable recovery (99.14%) and intra- and
inter-day precision (RSD 1.41%). Besides that, the method showed robustness to small modifications in the
analytical conditions (pH, mobile phase, temperature, flow and detector). Conclusion. Results confirm that the
proposed method can used for to quantify this drug in routine analysis and stability studies in the presence of
degradation products and synthesis impurities.
Financial Support
Projeto Universal CNPq (Processo 457629/2014-2) and UNIPAMPA (first author's masters fellowship)
116
DEVELOPMENT AND VALIDATION OF LC METHOD AND MICROBIOLOGICAL ASSAY
TO DETERMINE DANOFLOXACIN IN INJECTION SOLUTION
RENATA MEDEIROS HILGERT
EVERSON WILLIAM FIALHO CORDEIRO
CHEILA DENISE OTTONELLI STOPIGLIA
Laboratory de Microbiology, University Federal of Pampa - UNIPAMPA, Uruguaiana, RS, Brazil
Introduction. Danofloxacin is a synthetic fluoroquinolone, analogous to ciprofloxacin, available in
pharmaceutical market for veterinary use as injectable solution. This drug is used in the treatment of diseases
related to the respiratory and gastrointestinal tracts of birds, pigs and cattle, with a broad action spectrum against
Gram negative and Gram positive. The literature researched shows some physicochemical methods to quantify
this drug, but microbiological studies are limited. Then the objective of this study was to develop and validate a
microbiological assay by agar diffusion and a liquid chromatography method to quantify danofloxacin using forced
degradation studies to simulate the formation of degradation products. Methods. For the microbiological assay
were used Staphylococcus epidermidis as test microorganisms and danofloxacin standard solutions at 5, 10 and
20 µg/mL. The chromatographic separation was performed in an Ascentis® column (150 x 4.6 mm, 5 µm) using a
Shimadzu Prominence® liquid chromatograph. The mobile phase used was composed of a mixture of 0.3%
triethylamine (pH 3.0) and acetonitrile (80:20, v/v), at a flow-rate of 1.0 mL min-1. The determination of drug was
performed at 283 nm using photodiode-array detection. Linearity, precision, accuracy, robustness, and specificity
were performed to validate the analytical methods. Results and Discussion. The methods showed a suitable
linearity, precision (intermediate precision and repeatability), accuracy, and robustness. However, the
microbiological method did not show specificity to quantify the drug in the presence of the photodegradation
product. Conclusion. These results suggest that the photodegradation products exhibit biological activity, limiting
the quantification of the drug in stability studies using the microbiological assay However, the liquid
chromatography method showed suitable for routine analysis and stability studies of danofloxacin.
Financial Support
PDA UNIPAMPA
Acknowledgments
PDA UNIPAMPA
117
VALIDATION OF A CAPILLARY ELECTROPHORESIS METHOD FOR THE ASSAY OF
DOXYCYCLINE HICLATE SUPPOSITORY
Suelen Leticia Burin
Ana Paula Christ
Introduction: Doxycycline hiclate (DOX) is an antibiotic from the tetracycline class used in human and
veterinary patients. Our group developed a suppository aiming the treatment of infections in pets. The use of
suppository has advantages in animals because of the ease of administration and the possibility of fewer adverse
effects when compared to oral administration. The objective of this work was the validation of a method to
determine DOX in the developed suppositories. Methods: The assay method was performed in an Agilent 7100
equipment, with the following conditions: running buffer composed by a mixture (80:20) of solution A and B (A,
sodium carbonate and EDTA, 25 mM + 5 mM, pH 10.6, and B, acetonitrile), a fused silica capillary of 40 cm x 50
mm (effective length x inner diameter), kept at 24°C, voltage of 25 kV and detection at 260 nm. The sample used
was suppositories containing a DOX quantity equivalent to 50 mg of doxycycline. The method was validated
according to the ICH Guideline. Results and discussion: The method showed linearity (r=0.999) in the range of 20
to 160 µg/mL with no significant linearity deviation and linear regression (p < 0.05). The specificity was
determined by stress test, and the DOX residual content were: 95.3% (HCl 0.1 M/26 h), 81.6% (H2O2 3%/5h),
76% (NaOH 0.1 M/14h), and 88.9% (40°C/72h). Although degradation, DOX peak purity was observed in all
conditions. The robustness was evaluated thought factorial test 23. No influence of single factor or combination of
them was observed in the method response. Precision was evaluated by intraday (RSD values of 1.84% and
1.21%) and inter-day assays (RSD=1.48%, n=12). The accuracy was inferred based on compliance of linearity,
specificity and precision parameters. Conclusions: The results demonstrated that the assay is efficient for the
determination of DOX in the developed suppositories. Financial Support: Capes and
FIT-UFSM.
Acknowledgements: Cristalia Brazil for providing DOX.
Financial Support
FIT/UFSM
Acknowledgments
Cristalia Brazil
118
OPTIMIZATION OF ANALYTICAL CONDITIONS FOR GC ASSAY OF THE MAIN
COMPOUNDS OF Aniba canelilla (H.B.K.) MEZ OIL IN SKIN MATRICES
Tainá Kreutz
Graduate Program in Pharmaceutical Sciences, Federal University of Rio Grande do Sul, Porto Alegre, RS, Brazil
Juliane Freitas Bica
Letícia Grolli Lucca
Renata Pereira Limberger
Valdir Florencio da Veiga Junior
Chemistry Department, Federal University of Amazonas, Manaus, AM, Brazil
Introduction: Aniba canelilla (H.B.K.) Mez is an aromatic plant of the Amazon region, which presents 1nitro-2-phenylethane (NF) and methyleugenol (ME) as main compounds. The oil presents anti-inflammatory
activity and has a historical evidence of topical use. However, the skin retention/permeation of its main
compounds is not known up to date. Therefore, this study aims to develop a bioanalytical method using solid
phase microextraction (SPME) gas cromatography (GC) in headspace mode (HS) for the direct assay of both
compounds after in vitro permeation studies of an oil-based emulsion under development by our research group.
Methods: A HS-SPME parameters were optimized to extract the main compounds using a Box Behnken 3³
factorial design (Minitab 17®). The two main compounds were analyzed in the receptor fluid, in tape strips
(stratum corneaum) and skin layers (epidermis and dermis) by GC with Flame Ionization Detector (GC-FID)
(Shimadzu 2010) after the skin permeation study on Franz cells mounted with porcine skin (VhTex®) (n=5).
Results and Discussion: According to the optimization study, the best conditions for extraction were 21 minutes,
53ºC and addition of NaCl 5% (w/v). The analysis of skin and fluid samples showed a great permeation of 1-nitro2-phenylethane and methyleugenol since 42.81±55.08 µg/ml (NF) and 2.01±1.24 µg/ml (ME) were found in the
receptor fluid. The compounds retention was in the order dermis >> epidermis >> stratum corneaum, as follows:
179.56±35.15 µg/cm² (NF) and 125.44±48.79 µg/cm² (ME); 26.11±11.00 μg/cm² (NF) and 44.04±19.95 μg/cm2
(ME); 5.52±4.63 µg/ml (NF) and 1.89±1.78 µg/ml (ME), respectively. Conclusions: The results were found
promising due to the high retention of the volatile compounds in the dermis, which is the skin layer where cells
and inflammatory mediators are located.
Financial Support
Financial support from CNPq/Brazil and master scholarship from CNPq/Brazil
119
CO-ENCAPSULATION OF QUININE AND CURCUMIN IN POLYMERIC NANOCAPSULES:
DEVELOPMENT AND CHARACTERIZATION
Tamara Ramos Maciel
Luana Roberta Michels
Kelle Velasques
Daiana Silva de Ávila
Sandra Elisa Haas
Introduction. Malaria is caused by intracellular protozoan called Plasmodium. Brazil is responsible for
52% of malaria cases and 59% of deaths in America. Quinine (QN) is the drug of choice for the treatment of
chloroquine-resistant malaria, but it has serious adverse effects such as cardiotoxicity, neurotoxicity and
hypoglycemia. The curcumin (CUR) is extracted from Curcuma longa and has antimalarial activity, but it has low
aqueous solubility, low bioavailability and susceptible to photodegradation. In order to improve the antimalarial
activity and avoid the main problems for these drugs, nanocapsules (NC) become promising to co-encapsulation
of QN and CUR. The aim of this work was to develop and characterize three formulations designed as NC-QN
(QN-loaded NC), NC-CUR (CUR-loaded NC) and NC-QN-CUR (co-encapsulation of QN and CUR-loaded NC).
Methods. Formulations were developed by interfacial deposition method of pre-formed polymer. The particle
diameter was determined by laser diffraction technique (Mastersizer 2000 Malvern®), the zeta potential was
determined by electrophoretic migration technique, the pH values was determinate with potentiometer (HANNA®),
the drug loading and encapsulation rate were quantified by HPLC-PDA. Results and discussion. The particle size
showed nanometric (NC-QN = 376 nm, NC-CUR = 406 nm and NC-QN-CUR = 435 nm). The zeta potential for
all formulations was negative. The pH values ranged from 6.8 to 8.0 for all suspensions. The drug content were
close to the theorical concentration. The encapsulation rate for NC-QN, NC-CUR and NC-CUR-QN were 68%,
97%, 90% (QN) and 97% (CUR), respectively. Conclusions. With these results, we demonstrate that is possible
to develop nanocapsules entrapping QN and CUR with appropriate physical and chemical characteristics.
Financial Support
FAPERGS
Acknowledgments
FAPERGS
120
()BORNEOL / βCYCLODEXTRIN HOSTGUEST SYSTEM: PHYSICOCHEMICAL
CHARACTERIZATION AND DISSOLUTION BEHAVIOUR
TATIANNY DE ARAÚJO ANDRADE
Federal University of Sergipe
PAULA DOS PASSOS MENEZES
BRUNO DOS SANTOS LIMA
IGOR ARAÚJO SANTOS TRINDADE
ISABELLA GONÇALVES MATOS
LUCINDO JOSÉ QUINTANS JÚNIOR
LUIZA ABRAHÃO FRANK
University of Rio Grande do Sul
SÍLVIA STANISÇUASKI GUTERRES
University of Rio Grande do Sul
ADRIANO ANTUNES DE SOUZA ARAÚJO
MAIRIM RUSSO SERAFINI
Introduction: The (-)-borneol is a monoterpene alcohol present in the essential oils of several medicinal
plants. With the perspective of conserving the therapeutic potential and improve the solubility, this study has to
perform the inclusion of this compound in βcyclodextrin (βCD). Methods: The samples were prepared with βCD
by physical mixture (PM) and slurry complexation (SC) methods. Subsequently, these samples were analysed by
thermogravimetry (TG) and scanning electron microscopy (SEM) as molecular inclusion. Furthermore, an
analytical method was developed and validated by high-performance liquid chromatography (HPLC) as per the
determinations of International Conference on Harmonization guidelines (ICH). After, in vitro dissolutions tests
were performed using capsules containing 10 mg of (-)-borneol free, PM and SC samples. Thus, water was used
as dissolution means and the test was maintained during six hours in a water bath and at certain time intervals (5360 minutes) aliquots were collected and quantified by HPLC analysis. Results and discussion: From the
obtained results the SC sample showed better profile of molecular inclusion than PM. In the second step of TG
analysis (140-280 ºC) SC lost 3.9% of (-)-borneol complexed while PM lost only 0.3%. In addition, the SEM
pictures showed the large crystal size of PM, similar to βCD sample, and SC exhibited smaller crystals with
different morphology. In the HPLC validation, all parameters analyzed showed significant results, with relative
standard deviation less than 15%, R2 0.9963. The amount of (-)-borneol dissolved was observed that free drug
dissolved 27%, PM 28.5% and SC 90%. The PM particles showed low complexing capacity, which was
confirmed by the dissolution test, where by this method only drug free was dissolved. Conclusion: The SC
method showed better efficient complexation and dissolution profile performed a good alternative for oral route
administration.
Financial Support
The authors are grateful to CAPES, CNPq, FAPITEC/SE for financial support and fellowships.
121
Investigation of adulterants in dietary supplements by Ion Exchage Chromatography
with Conductometric Detection
Thais Ramos Dal Molin
Gabriela Camera Leal
Larissa Sabo Müller
Diana Tomazi Muratt
Carine Viana Silva
Leandro Machado Carvalho
The dietary supplements are products suitable for athletes in order to replenish nutrients and other
substances lost over the intense physical activities, or else to supplement a diet when this is insufficient. In
Brazil there is no official legislation for dietary supplements. Sometimes the same product can be regulated by
more than one resolution, as result of this many adulteration cases in these products has been described in the
literature. The Ion Exchange Chromatography with Conductivity Detection (IEC-CD) consists in the electrostatic
interaction between the interesting analytes and binding sites of the column combined with a universal detector,
and it has been considered an analytial method in potential to investigation of adulterations. Thus, this work aimed
the optimization, validation and application of a screening method for the investigation of presence of
amfepramone, femproporex, sibutramine, bisacodyl and amiloride in dietary supplements samples obtained from
Brazilian websites. The method was developed with a cation exchange column Metrosep C4 100/4.0. The mobile
phase consisted of 1.8 mM HNO3 containing 2% of acetonitrile, with flow rate of 0.9 mL.min-1 with conductivity
detection without suppression. The limits of detection (LOD) varied from 1.01 up to 3.62 mg.mL-1, whereas the
limits of quantification (LOQ) were in the range of 1.48 up to 8.72 mg.mL-1. The proposal method was applied in
78 solids samples of dietary supplements, and two showed be suspected of adulteration. In one of the sample
was detected 9.877 ± 0.05 mg.g-1 of sibutramine and another one 8.076 ± 0.01 mg.g-1 of bisacodyl. The method
presented be linear, specific, selective and applicable for large number of samples. The possible presence of
these adulterants suggests more investigations about the real information described on the labels of dietary
supplements, pre-market safety and efficacy tests and manufactor quality control according current legislation.
Financial Support
CAPES
Acknowledgments
The authors wish to acknowledge the financial support given by the Brazilian foundations CNPq, FAPERGS and CAPES
122
IN VITRO RELEASE OF SIMVASTATIN FROM CHITOSAN POLYMERIC FILMS
Vandiara Martins Moreira
Amaro César Lima de Assis
Natan Emanuell de Sobral e Silva
Pedro Gomes da Silveira Neto
Yuri Basílio Gomes Patriota
Fellipe Fernandes Santos
Ana Cláudia Gonçalves dos Santos
Bruna Pereira da Silva
Polymer systems drug release stage results from the influence of dissolution mechanisms such as
diffusion and erosion of the polymer. This complex release mechanism depends on the exposure time in water,
because there is a transition between the dry and malleable state. The purpose of this study was to evaluate the
in vitro kinetics release of simvastatin (SIMV) from chitosan (CTS) polymeric film for topical use. Four types of
CTS polymeric films containing SINV were used [two produced with CTS from Sigma-AldrichTM (10 and 20 mg
of SIMV) and two produced CTS from PhytomareTM (10 and 20 mg of SIMV). For in vitro release, a Franz
apparatus was used, in which the receiver compartment has been filled with phosphate buffer 0.01M pH 7.0 and
the film was employed as a selective membrane and giver compartment. The SIMV quantification was performed
in UV spectrophotometer (238 nm). From the release profiles of SIMV, four kinetic models were applied to the
system (zero order, first order, Higuchi and Korsmeyer-Peppas) and a correlation coefficient (r) was used to
identify which kinetic model was more suitable for describing the drug release. All films of CTS polymeric film
containing SINV showed similar release profiles.They showed a notable control of drug release along the 24
hours, regulated by an anomalous transport, reported through the Korsmeyer-Peppas kinetic model. Therefore, it
can be suggested that the anomalous release mechanism occurs as a result of the combined action of the
processes of diffusion and erosion of the polymer matrix, what results in the kinetic release of SIMV from the
films for this model.
Acknowledgments
CAPES and CNPq.
123
DISSOLUTION METHOD FOR DELAPRIL AND MANIDIPINE COMBINATION TABLETS
BASED ON ABSORPTION PROFILE OF MANIDIPINE
Vítor Todeschini
Maximiliano da Silva Sangoi
Gustavo Krumel Goelzer
Jaison Carlosso Machado
Nadia Maria Volpato
Introduction: The association of delapril (DEL) and manidipine (MAN) is an important alternative to
arterial hypertension treatment. However, there is no available dissolution method for simultaneous evaluation of
drugs in pharmaceutical formulations. Objective(s): The aim of the present study was the development and
validation of a dissolution method for DEL and MAN combination tablets, using a simulated absorption profile
based on in vivo data for MAN. Methods: After sink conditions and drugs stability evaluations, the suitable in vitro
dissolution profile for this formulation were achieved using 900 mL of pH 3.2 citrate buffer at 37 °C ± 0.5 °C as
dissolution medium and USP apparatus 2 (paddle) at 75 rpm. All samples were analyzed by a validated liquid
chromatography method. Results and Discussion: Under these conditions a significant linear relationship between
the absorbed (calculated by deconvolution approach) and dissolved fractions of MAN was obtained (R = 0.997)
and a level-A in vivo-in vitro correlation could be established for this drug. Validation parameters for dissolution
methodology such as the specificity, linearity, accuracy and precision were also evaluated according to
international guidelines, giving results within the acceptable range. Conclusions: Therefore, the proposed
dissolution conditions can be applied for the simultaneous release analysis of DEL and MAN from the solid
dosage form, contributing to improvement of the quality control of pharmaceutics and to minimize the number of
bioavailability studies.
Financial Support
CAPES/Brazil
Acknowledgments
CAPES/Brazil for the financial support.
124
125
AMBULATORY HYPERTENSION IN BLACKS IN SOUTHERN BRAZIL
Ana Claudia Colombo de Oliveira
Patricia Maurer
Lyana Feijo Berro
Jacqueline Costa Escobar Piccoli
Introduction: The black population presents a high cardiometabolic risk and prevalent hypertension. The
best way to evaluate the effectiveness of treatment it is by ambulatory blood pressure monitoring (ABPM) 24
hours. Therefore, the objective of the study was to evaluate the prevalence of hypertension in a sample of the
black population of southern Brazil and the effectiveness of drug treatment. Methods: The ethics committee in
research of the Federal University of Pampa approved the project (977.827). Participated black self-declared
individuals that signed the consent form and answered a structured questionnaire. Blood pressure measurements
were obtained. Later, the subjects were classified as hypertensive x nonhypertensive. The hypertensive group
was invited to participate in an evaluation with the ABPM and we use the V Brazilian Guidelines to classify
patients. Results and discussion: Participated 203 people declared themselves black, average age 46.7 years old,
most female (162, 79.8%). 113 (54.3%) were hypertensive. Of this, 60 (53.1%) had uncontrolled hypertension and
53 (46.9%) had blood pressure controlled. After the initial interview, 31 hypertensive made the evaluation with the
ABPM. The results of 24-hour monitoring indicated that 54.8% had ambulatorial hypertension (above the limits
recommended by V guideline) and most had high blood pressure during sleep (nocturnal dipping). Conclusion: The
Afro-descendant population studied presents a high rate of ambulatorial hypertension indicating difficulty in blood
pressure control despite the use of more than one antihypertensive class. Pharmacotherapeutic interventions are
needed to optimize the causes of not controlling blood pressure in the black population.
Financial Support
CNPq
Ethical approval
977.827
126
REDOX STATUS IN MUCOPOLYSSACHARIDOSIS IVA PATIENTS UNDER ENZYME
REPLACEMENT THERAPY
Bruna Donida
Biological Science: Biochemistry Postgraduate Program, University of Rio Grande do Sul, Porto Alegre, RS, Brazil.
Desirèe Padilha Marchetti
Carlos Eduardo Diaz Jacques
Caroline Paula Mescka
Medical Genetics Service, Clinical Hospital of Porto Alegre, Porto Alegre, RS, Brazil.
Roberto Giugliani
Medical Genetics Service, Clinical Hospital of Porto Alegre, Porto Alegre, RS, Brazil.
Carmen Regla Vargas
Introduction: Mucopolysaccharidosis type IVA (MPS IVA) is a rare lysosomal storage disorder caused
by a deficiency in the N-acetylgalactosamine-6-sulfatase enzyme, causing an accumulation of
glycosaminoglycans (GAGs) in body fluids and lysosomes of many organism´s cells. Considering the involvement
of oxidative damage in other MPSs, the aim of this study was to evaluate redox status and GAGs levels in
patients with MPS IVA under enzyme replacement therapy (ERT). Methods: This study was approved by The
Ethics Committee of the Clinical Hospital of Porto Alegre, Brazil (No.13-0246). Heparinized blood and urine were
obtained from patients (n=17) and from healthy age-matched controls (n=10-15). All MPS IVA patients were
receiving ERT treatment (Vimizim® 2mg/kg) every week by intravenous infusion, for 32 weeks approximately.
Reduced glutathione (GSH), a non-enzymatic antioxidant, was measured by a fluorescence assay in
erythrocytes. Antioxidant enzymes superoxide dismutase (SOD) and glutathione peroxidase (GPx) were
measured in erythrocytes by commercial kits. Concentration of plasma sulfhydryl groups and urinary GAGs levels
were determined by spectrophotometric techniques. Results and Discussion: MPS IVA patients presented a
reduction of antioxidant defense levels, assessed by a decrease in GSH content when compared to control group.
SOD activity was significantly increased in patients, while there was no significant difference in GPx activity
between patients and controls. It was verified a decreased sulfhydryl content in MPS IVA patients compared to
controls, suggesting protein oxidative damage. GAGs levels were still high in patients compared to the control
group. The data presented propose that disturbances in redox status occur in MPS IVA patients even under ERT.
Conclusions: Considering the results, supplementation of antioxidants in combination with ERT can be a
promising therapeutic approach for MPS IVA.
Financial Support
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq).
Ethical approval
This study was approved by The Ethics Committee of the Clinical Hospital of Porto Alegre,
Brazil (No.13-0246)
127
EVALUATION OF ANTINEOPLASTIC POTENTIAL OF NOVEL OXAZOLIDINE
DERIVATIVES: CYTOTOXICITY AND MODULATION IN EXPRESSION OF APOPTOSISRELATED GENES IN ACUTE T-CELL LEUKEMIA
Caio Gomes de Barros Martins
Laboratório de Imunomodulação e Novas Abordagens Terapêuticas - LINAT/UFPE
Wanessa Layssa Batista de Sena
Laboratório de Imunomodulação e Novas Abordagens Terapêuticas - LINAT (UFPE)
Júlia Furtado Campos
Marina da Rocha Galdino-Pitta
Laboratório de Planejamento e Síntese de Fármacos - LPSF (UFPE)
Ivan da Rocha Pitta
Maria do Carmo Alves de Lima
Michelly Cristiny Pereira
Intro: Concern about cancer has risen since the treatment is yet not completely effective. Toxicity and
development of resistance to the drugs in clinical use are additional problems that became targets for international
research. Therefore, an advance in studies involving new potential antineoplastic drugs such as the oxazolidine
derivatives has become crucial. Methods:Jurkat cells were cultured in complete RPMI medium and treated with
10, 25, 50 and 75μM of each of the following derivatives: NB-2, NB-4, NB-5 and NB-6. Their cytotoxic potential
was assessed by the MTT assay. Gene expression of BID, p53 and BCL-2 was evaluated by qRT-PCR and
normalized to 18S values. In order to do that, cDNA was synthesized using RNA extracted from cells treated
with IC50 doses, determined by the previous assay. Statistical analysis of the two independent experiments was
performed using t-student test and differences were considered significant when p<0.05. Results/Discussion: All
oxazolidine derivatives showed cytotoxic activity against Jurkat cells. The IC50 for NB-2, NB-4, NB-5 and NB-6
were calculated as 24.18, 15.19, 22.54 and 28.99μM, respectively. The NB-6 was able to significantly up-regulate
genes such as BID (p=0.0092) and BCL-2 (p=0.0013). The same was observed for the NB-5 which was able to
induce expression of p53 (p=0.0171). Increased levels of BCL-2, an antiapoptotic gene, might be explained as a
negative feedback response of tumour cells to increasing apoptosis stress induced by activation of both BID and
p53. The treatment with remaining derivatives did not show statistically differences in expression of p53, BID or
BCL-2. Conclusion: All oxazolidines derivatives showed cytotoxic activity against Jurkat. In addition, NB-5 and
NB-6 were able to modulate the expression of apoptosis-related genes indicating the involvement of BID and p53
in cell death. Nevertheless, further studies must be performed in order to thoroughly interpret cell death
mechanisms.
Financial Support
UFPE, CNPq, INCT_IF
128
ANTIBACTERIAL ACTIVITY OF MUSA SPP. PEEL EXTRACTS
Camila de Oliveira Melo
Roxana Costa Nobrega Acioli
Pablo Queiroz Lopes
Edeltrurdes de Oliveira Lima
Aratã Oliveira Cortez
Anna Emannuela Medeiros de Brito
Mysrayn Yargo De Freitas Araujo Reis
Elquio Eleamen Oliveira
Introduction: Due to the relevance of the knowledge of traditional folk medicine on the use of banana
peel for therapeutic purposes: antimicrobial, antioxidant, anti- inflammatory, among others, the present study
evaluated the antimicrobial activity of banana peel extracts to predict the viability to develop a topical formulation
based on banana peel extract for the treatment of microbial infections. Methods: Determination of Minimum
Inhibitory Concentration (MIC) of two extracts (ethanolic extract 70% and acetone:water (1:1) extract) was
performed by broth microdilution technique using microdilution plates containing 96 wells-bottomed "U" (all tests
were made in duplicate). In each well of the plate was added 100 µL CN (bacteria) doubly concentrated.
Subsequently, 100µL of sample, also doubly concentrated, were added into the first wells row of the plate and by
means of a serial dilution of two fold were obtained the concentration of 1024 µg/mL to 32 µg/mL of the extracts.
Finally, it was added 10uL of the inoculum of the microorganisms in the wells. The follow strains were used in the
trials of antibacterial activity in vitro of the extracts: Staphylococcus aureus ATCC-25923 and LM-177, S.
epidermidis ATCC-12228, P. aeruginosa ATCC-25853 and P-03, Bacillus subtillis ATCC- 6633, Echerichia coli
ATCC- 10436 and EC-12. Results and Discussion: Bacterial strains of Staphylococcus aureus ATCC 25923 and
LM- 177, P. aeruginosa ATCC- 25853 and P03 were sensitive to extracts obtained from maceration of banana
peels in acetone:water and ethanol 70 %. The ethanolic extract of banana peels showed the best antimicrobial
activity (256 µg/mL). Moreover, the acetone extract: water showed MIC of 1024 µg/mL. Conclusions: Both
extracts showed activity against all bacteria tested in this work, with better results for the ethanolic extract,
allowing further studies in the development of a formulation based on the banana pell with antibacterial activity.
129
PHENOTYPIC EXPRESSION OF BIOFILM IN ENTEROBACTERIA AND ANTIBACTERIAL
SUSCEPTIBILITY
Clarissa Figueredo Braga
Narcisa Caroline de Oliveira Silva
Ana Beatriz Sotero Siqueira
Among the virulence factors, biofilm formation has been widely cited as a promoting agent of
pathogenicity in clinical complications. Besides facilitating the production of other virulence factors, this type of
organization provides antimicrobial resistance to bacteria, which takes to limited or absent therapeutic options and
is a risk factor for persistence and worsening of an infection. Thus, the aim of this study was to analyze the
phenotypic expression of biofilm in bacteria from the family Enterobacteriaceae and to verify in vitro the
susceptibility of bacterial isolates to commercial drugs. For the experiments, 19 isolates of Escherichia coli and
19 of Klebsiella pneumoniae were used, which were provided by the Bacteriology Department of the Clinical
Hospital - UFPE. Biofilm formation was determined by absorbance test in polystyrene microplate, and
susceptibility tests were performed with 13 commercial drugs according to the procedures adopted by the Clinical
and Laboratory Standards Institute. For each species, it was found that 84.2% of the isolates formed biofilms.
Regarding susceptibility, 6 isolates of E.coli and 11 of K. pneumoniae were resistant to more than half of the
drugs used, and presented one isolated resistant to all, with penicillins, quinolones and cephalosporins as
predominant groups. The relation between virulence factor expression and antibiotic susceptibility for isolates that
expressed biofilm and were also resistant to 50% of the drugs was checked. That relation was already found in
many clinical studies. Our results showed that both species expressed a similar potential in forming biofilm, and
showed resistance to the drugs mainly by Klebsiella pneumoniae. All resistant isolates had biofilm formation,
which indicates it as an actual preponderant factor for the resistance. This reveals the importance of the
continuous search for new therapeutic strategies against these bacteria.
130
FUNGICIDAL ACTIVITY OF MILTEFOSINE AGAINST Paracoccidioides brasiliensis AND
Paracoccidioides lutzii ISOLATES.
cristina de castro spadari
USP
Diego Conrado Pereira Rossi
USP
Carlos Pelleschi Taborda
USP
Kelly Ishida
USP
Paracoccidioidomycosis (PCM) is a systemic and endemic mycosis in Latin America caused by the
dimorphic fungus Paracoccidioides spp. The treatment of PCM is based in the administration of itraconazole
(ITZ), amphotericin B (AMB) or combination of sulfamethoxazole (SMX) and trimethoprim (TMP); however, it is
long and can last from six months to two years and relapses may occur. Thus, the search for new therapeutic
options is necessary for the treatment of this mycosis. Miltefosine (MLT), an analogue of alkylphospholipids, has
displayed a good antifungal activity against different species of yeast and filamentous fungi. The aim of this study
is to evaluate the activity of MLT on Paracoccidioides brasiliensis and Paracoccidioides lutzii isolates. The
viability was determined by the time-kill assay. Yeasts of P. brasiliensis (Pb18) or P. lutzii (Pb01) (1 x 105 cfu/ml)
were treated with different concentrations of MLT (0, 0.5xMIC, MIC, 2xMIC, 4xMIC and 8xMIC) in MVM medium
for 1, 3, 5, 7 and 14 days at 37 ºC. At each time, treated and untreated yeasts were diluted in PBS and 100 µl
were plated on the BHI agar medium supplemented with 4% of fetal calf serum and 5% of P. brasiliensis culture
filtrate at 37ºC for 14 days before colony forming unit (c.f.u.) counting. For both species, concentrations above
MIC value of MLT (>1 µg/ml) were able to reduce the viable yeasts to no detectable levels by the c.f.u. counting
after 1 day of the drug exposition. Yeasts from Pb18 and Pb01 were eliminated when treated with 1 µg/ml MLT
after 5 and 7 days of incubation, respectively. Sub-inhibitory concentration of MLT (0.5 µg/ml) inhibited about 50%
of yeast growth of Pb01 after 14 days of incubation in relation to the untreated yeasts; in contrast, Pb18 growth
was not inhibited by the same MLT concentration. In conclusion, time-kill assays revealed the fungicidal activity
of MLT and more studies should be conducted for possible use in the treatment of the PCM.
Financial Support
CAPES, CNPq, FAPESP
131
IN VITRO EVALUATION OF MCF-7 CELLS SURVIVAL EXPOSED TO HYPOXIA AND/OR
NUTRIENT DEPRIVATION MEDIATED BY GAL-3
Edso Henrique Batista Barros
Federal University of Pernambuco
Maíra Galdino da Rocha Pitta
Ivan da Rocha Pitta
Antônio Félix da Silva Filho
Introduçtion: Cancer represents one of the leading causes of death in the world. Breast cancer
microenvironment features low oxygen tension and nutrients starving, Nearby tumor cells adapt itself to hypoxia
via HIF1α wich induces the transcription of several genes, including Galectin-3. Thus, this work aims to
characterize the in vitro model of Gal-3 on survival of neoplastic cells from breast on this adverse environment.
Methods: MCF-7 cells were cultured in RPMI 1640 supplemented with 10% fetal bovine serum (FBS) incubated in
a humid atmosphere of 5% CO2 at 37°C. Cells were subjected to the conditions of study N10 (normóxia with
10% of FBS); N1 (normoxia with 1% SFB); H10 (hypoxia with 10% FBS) and H1 (hypoxia with 1% FBS). The
oxygen deprivation was established in hipoxic chamber in 24 and 48 hours. Cell death was evaluated in flow
citometry staining with PI. The analysis of Gal-3 transcripts was perfomed by quantitative real time PCR in and
western blotting to investigate the expression and translation of Gal-3 to the intracellular medium. Results and
Discussion: In MCF-7 cells, hypoxia combined with nutrient deprivation (H1) demonstrated the highest death rate
(47% ± 0.1), followed by nutritional deprivation (18% ± 11.1) in 48 hours. Analyzing the expression of transcripts of
Gal-3 under the conditions of study proposals, N1 and H1 determined the highest levels of expression of Gal-3:8.6
and 8.2, respectively, at the time of 24 hours. The western blotting concluded increased amount of Gal-3
expressed in MCF-7 cells, where H1 and N1 showed increase of 4 and 2.3 times, respectively in 48 hours. The
level of transcripts and proteins of Gal-3 is increased in the conditions N1 and H1, suggesting the participation of
Gal-3 metabolic adaptation and survival of these cells in the hostile environment. Conclusion: The results indicate
that Gal-3 is modulated in transcriptional and protein levels in N1 and H1 conditions.
Financial Support
CNPq
132
TIAZACRIDINE DERIVATIVES INDUCED CYTOTOXICITY AND INHIBITED CELL
MIGRATION IN BREAST CANCER MCF-7 CELLS
Flaviana Alves dos Santos
Laboratory for Immunomodulation and New Therapeutic Approaches, Federal University of Pernambuco (UFPE), Recife, PE, Brazil.
Maria do Carmo Oliveira Lima
Laboratory for Planning and Drug Synthesis, Federal University of Pernambuco (UFPE), Recife, PE, Brazil.
Marina Galdino da Rocha Pitta
Ivan da Rocha Pitta
Laboratory for Immunomodulation and New Therapeutic Approaches, Federal University of Pernambuco (UFPE), Recife, PE, Brazil.;
Breast cancer is the most incident cancer in women worldwide. Therefore, design and development of
new bioactive agents has become important. The molecular hybridization strategy involves the integration of two
pharmacophoric groups with different mechanisms of action in the same molecule. Thiazacridine derivatives are
formed by the thiazolidine and acridine ring junction which are known to suppress tumour growth and intercalate
with DNA. MCF-7 cells were cultured in RPMI supplemented with 10% FBS. Cytotoxic activity was performed by
the MTT assay. The compounds tested were LPSF/AC-99, LPSF/AC-119 and LPSF/AA-29 in concentrations
between 1 µM and 75 µM. We also performed in vitro scratch assay with cells treated with complete medium,
0.1% DMSO and LPSF/AA-29 (IC50 and IC25 values). Cells were photographed in t=0h and 48h. Data of
migration assays were obtained in relation to percentage of DMSO treated cells. We observed that only the
tiazacridine LPSF/AA-29 derivative reduced cell viability, with IC50 of 40.76µM and doxorubicin was 6.51
µM(positive control). The derivatives LPSF/AC-99 and LPSF/AC-119 presented IC50 >75µM. We observed that
treatment with LPSF/AA-29 during 48h is highly effective against MCF-7 cell motility in both concentrations of
IC50 (p=0.014±5.21) as the IC25 (p=0.032±3.65).Only LPSF/AA-29 has the 5-bromo-indole ring in their chemical
structure. This chemical grouping may have increased cytotoxic potential of this derivative and some articles
suggest that it increases the cytotoxic activity against bladder, colon rectal and breast cancer. Cell invasion, entry
and exit from the circulation and colonization at the distal sites, known as metastasis, is the cause of increased
mortality in patients with cancer. To find new drugs that inhibit migration and invasion is a major on going effort for
cancer therapeutics. The derivative thiazacridine LPSF/AA-29 inhibited cell proliferation and migration of MCF-7
cells.
Financial Support
CAPES, CNPQ, FACEPE
133
ENTEROPARASITOSIS CONTROL IN CHILDREN OF THE DE SAINT FRANCIS ASSIS
CITY - RS
Flaviana Zanini Marin
Faculty of Pharmacy, Universidade Regional Integrada do Alto Uruguai e das Missões-URI, Santiago, RS, Brasil.
Saionara Benvenhú Machado
Nilce Ione Vielmo
Mirian Caetano
Thaylise Vey Parodi
Parasitic infections are caused by intestinal parasites, which affect mainly developed / underdeveloped
countries. These parasites are among the pathogens most often found in humans, affecting mainly children and
adolescents. Their proliferation of media are basically related to sanitation, climate, nutritional status of individuals
and the level of hygiene. This study aimed to evaluate the prevalence of parasitic infections in 61 children aged 0
to 5 years old, assisted by the Family Health Strategies (ESFs) in the city of St. Francis of Assisi, under the
approval of the Research Ethics Committee through number 1,124,706. Once approved, the guardians of the
children signed the consent form and then answered a socio-cultural questionnaire, which through this we can
identify the key hygiene habits that people had. Is extremely important, because only by knowing the relevant
factors for transmission of these parasites, is that we can set goals, plan means of prevention and control of
parasitic diseases. Samples were analyzed macro / microscopically resulting in 53 samples negative for parasitic
infections and 8 samples. Of the positive samples, 6 corresponded to cysts of Giardia lamblia and Entamoeba
coli 2. After the analysis, health education actions were developed in order to discuss with the subject matters
related to the prevention, treatment and sanitation, providing greater understanding of them. Finally we conclude
by the results that the prevalence of parasitosis was significantly less than 13% from the total sample. This low
rate is associated to good daily hygiene and pharmaceutical care work provided during project execution. It is still
relevant to be implemented to population, educational measures in their daily lives, to improve awareness and
improvement in quality of life.
Financial Support
Acknowledgments
Os autores do presente trabalho são gratos pelo apoio financeiro da URI.
Ethical approval
Nº. 1124 .706.
134
ASSESSMENT OF THE ANTIMICROBIAL POTENTIAL FROM DIFFERENT EXTRACTS
FROM THE LEAVES OF THE Eugenia pyriformis Cambess.
Jéssica Aparecida Coelho
State University of Western Paraná (UNIOESTE)
Simoni Cristina Saling
Catholic University of Paraná (PUCPR)
Introduction: The decoction, in water, of the leaves of Eugenia pyriformis Cambess, also known as
uvaia can be useful in the control of hypertension, cholesterol lowering, inflammatory and infectious processes. It
also shows high antimicrobial and antioxidant capacity in vitro. Methods: The extracts ethyl acetate (EAE),
ethanol (EE) and hexane (SH) were obtained from the powder of the leaves of E. pyriformis by the addition of the
solvents at the proportion of 1: 5 (plant: solvent) and the aqueous extract (AE) at the proportion of 1:10 (plant:
water). The mixtures were into maceration or infusion up to 24 hours, filtered and lyophilized. The antimicrobial
activity evaluation was carried out by a comparative analysis of the inhibition halos (diffusion-disk), along with the
reference antibiotic discs. Three concentrations of the extract were tested: 6, 12 and 18 UI/mL, besides the
positive and negative controls. The statistical treatment was carried out by the analysis of variance (ANOVA)
followed by the Tukey test, considering a significance level of 5%. Results and Discussion: All four extracts'
concentrations used in evaluating potential antimicrobial against the tested organisms (Staphylococcus aureus,
Escherichia coli, Salmonella typhimurium, Pseudomonas aeruginosa e Streptococcus pyogenes) were able to
generate a halo growth inhibition of the microorganisms. In a general context, the statistical differences showed
that the extracts have greater antibacterial potential against Gram-positive than against Gram-negative bacteria.
Furthermore, the EA and the EAE were more effective against the growth of the tested organisms, they were, in
fact, more efficient than the antibiotic vancomycin 30 μg, and the penicillin G 10 μg alike. Conclusions: This
study contributes to justify the use of uvaia, in popular medicine, for the treatment of infections, once the tested
extracts showed antimicrobial potential against some of the major pathogens that affect humans.
Acknowledgments
Catholic University of Paraná (PUCPR)
135
EVOLUTION OF ENTEROPARASITOSIS FREQUENCY IN STUDENTS OF PONTA
GROSSA- PR (2008 - 2014)
Júlio César Miné
Universidade Estadual de Ponta Grossa (UEPG)
Priscilla de Brito Dória Jorge
The prevalence of Intestinal parasites requires constant monitoring, as it is responsible for many health
problems in individuals; especially children, who are the most affected - and their physical and intellectual
development can be compromised. The extension project: "Enteroparasitosis in children in the region of Ponta
Grossa - PR" exists for years in order to assess the frequency of intestinal parasites in schoolchildren from the
city of Ponta Grossa, PR, and promotes both technical education to students coursing Pharmacy and guidance to
the schoolchildren and their parents in parasitic diseases prophylaxis. The aim of this study was to compare the
frequency of intestinal parasites in Ponta Grossa students in the period between 2008 and 2014. The
parasitological analyzes were performed by the methods of Hoffman, Pons and Janer and Faust et al. 2096
examinations were performed, of which 26.38% had positive results for the presence of intestinal parasites. In
2008, the rate of positive tests was 30.48%; it rose to 42.64% in 2009 and since 2010 there were reductions in
the prevalence reaching 16.45% in 2012. In 2013, however, the rates returned to rise (22.04%) and in 2014 the
lower percentage was observed (13.00%). Protozoa and helminths most commonly found in these seven years
were Entamoeba coli, Giardia lamblia and Trichuris trichiura and Ascaris lumbricoides, respectively. The nonpathogenic protozoa are indicative of fecal contamination of the environment where the students are located. The
high frequency of positive fecal examinations in 2013 can be explained by the fact that several schools were
invited to participate in the project, which resulted in a greater number of parasitized schoolchildren, also because
they had never taken part of these project's activities before. It is necessary to monitor the schools' health
conditions through fecal examinations and implement procedures which will help, guide and aware the population.
Financial Support
Araucaria Foundation and PROEX / UEPG
Acknowledgments
Profa. Stella de Medeiros Ivahi badaró
Ethical approval
#501.171 /2013 COEP - UEPG
136
DEVELOPMENT OF A SENSOR PLATFORM BASED ON GOLD NANOPARTICLESPOLYANILINE HYBRID COMPOSITE FOR ELECTROCHEMICAL DETECTION OF
BRC/ABL FUSION GENE
Karen Yasmim Pereira dos Santos Avelino
Programa de Pós-Graduação em Bioquímica e Fisiologia, Universidade Federal de Pernambuco, Recife, PE, Brasil.
Isaac Aaron Morales
Programa de Pós-Graduação em Inovação Terapêutica, Universidade Federal de Pernambuco, Recife, PE, Brasil.
Norma Lucena Cavalcanti Licínio da Silva
Centro de Pesquisas Aggeu Magalhães (CPqAM), Fundação Oswaldo Cruz, Recife, PE, Brasil.
Celso Pinto de Melo
Laboratório de Polímeros Não-Convencionais, Departamento de Física, Universidade Federal de Pernambuco, Recife, PE, Brasil.
César Augusto Souza de Andrade
Laboratório de Biodispositivos Nanoestruturados, Departamento de Bioquímica, Universidade Federal de Pernambuco, Recife, PE, Brasil.
Maria Danielly Lima de Oliveira
Laboratório de Biodispositivos Nanoestruturados, Departamento de Bioquímica, Universidade Federal de Pernambuco, Recife, PE, Brasil.
Introduction: The genetic translocation between chromosomes 9 and 22, also called of BCR/ABL fusion
gene, is a leading biomarkers used for diagnosis of chronic myelogenous leukemia and acute lymphocytic
leukemia. Currently, the methods used for identification of this oncogene present some limitations. Thus, the
development of an effective molecular assay is essential for the identification of early-stage cancer. This study
has the objective of develop a biosensitive platform based on hybrid composite of gold nanoparticles (AuNps) and
polyaniline (PANI) for detection of BCR/ABL fusion gene. Methods: Cyclic voltammetry and electrochemical
impedance spectroscopy techniques were used to characterize the different steps of construction of the
biosensor. The cyclic voltammograms were obtained in a potential range of -0.2 to +0.7 V at a scan rate of 50
mV/s. The impedance spectra were recorded in a frequency range of 100 mHz to 100 KHz with a sine wave
potential of 10 mV. Biosensitive platform was obtained through the electrostatic immobilization of a DNA probe on
a gold surface modified with AuNpsPANI composite. Results and Discussion: Due to the synergism between the
physic-chemical properties of the AuNPs and PANI was possible to obtain a biocompatible nanostructured
platform with amplification of surface area. The interaction of the biosensor with different concentrations of
recombinant plasmid containing the BCR/ABL fusion gene resulted in changes in the values of amperometric
current and charge transfer resistance, indicating the biospecific recognition process. The biosensor showed high
sensibility, selectivity and a detection limit of 69.4 atomolar. Furthermore, this system presented excellent
analytical performance for detection of the oncogene in clinical samples of patients with leukemia. Conclusions:
Thus, the developed biosensitive platform can be an alternative tool for the molecular diagnosis of cancer and
monitoring of minimal residual disease.
Financial Support
CNPq, PROPESQ-UFPE e CAPES-Rede nanobio/ELINOR.
137
PROTEOMIC EVALUATION OF THE MECHANISM OF ACTION OF A NOVEL
OXAZOLIDINE DERIVATIVE IN SENSITIVE AND CHEMOTHERAPY-RESISTANT
LEUKEMIA CELL LINES
Lidiane Vasconcelos do Nascimento Carvalho
Júlia Furtado Campos
Luciana Pizzatti Barboza
Brazilian National Cancer Institute
Eliana Saul Furquim Werneck Abdelhay
Brazilian National Cancer Institute
Ivan da Rocha Pitta
Moacyr Jesus Barreto de Melo Rego
Introduction: Leukemias are diseases of progressive course in hematopoietic progenitor cells. The
widely used treatment combines several drugs that are associated to many side effects. This work aimed to
evaluate through proteomic analysis the mechanism of action induced by a novel oxazolidine derivative, in
sensitive and chemotherapy-resistant leukemia cells. Methods: Acute leukemia cell lines HL-60 and HL-60/MX1
(chemotherapy-resistant) were cultured and treated with the oxazolidine derivative LPSF/NB-2 at IC50
concentration in each cell line. After 48 hours, protein extraction was performed. Label-free quantitative proteomic
analysis was made using two-dimensional liquid chromatography coupled to a mass spectrometry. Results: In
proteomic analysis they were only considered modulation of proteins found in all experimental replicates of each
sample and with a modulation difference higher than twice differentially in HL-60 and HL-60/MX1 cells treated with
LPSF/NB-2 when compared to non-treated cells. After that, 330 proteins in sensitive cell line and 260 in resistant
cells were evaluated by their interaction and modulated pathways using MetaCoreD. Pathways involved in
cytoskeleton remodeling, cell cycle and regulation are differentially expressed in both cell lines. Proteins evolved
in apoptosis and survival pathways were modulated only in sensitive cell line, while epigenetic regulation of gene
expression were modulated only in resistant cells. Analysis of interaction networks indicates that both cell lines
express an important regulatory gene (c-Myc). Protein encoded by this gene plays a role in cell cycle progression,
apoptosis and cellular transformation. Conclusion: Oxazolidine derivative LPSF/NB-2 is capable to modulate the
expression of proteins and pathways involved in tumorigenesis.Further studies will allow the validation of these
proteins and the understanding about their participation in leukemia progression and resistance process.
Financial Support
FACEPE (Pernambuco State Research Support Foundation) and INCT_if (National Institute for Science and Technology in Pharmaceutical Innovation).
Acknowledgments
FECEPE, INCT_if, NUPIT, INCA.
138
EVALUATION OF GALECTIN-3 SECRESSION IN PANCREATIC CANCER CELL LINES
UNDER HYPOXIA AND/OR STARVING CONDITIONS
Lucas Batista Tavares
Ivan da Rocha Pitta
Antônio Félix da Silva Filho
Introduction: Therapy resistance and high aggressiveness characterize pancreatic cancer. These
malignancy features are associated with intratumoral hypoxia that leads to hypoxia-inducible factor stabilization
(HIF1α), which has influence in the proteomic profile of cancer progression. Galectin-3 (Gal-3) shows altered
expression in hypoxia and performs many functions in this microenvironment including angiogenesis and
metastasis promotion. This study evaluated the extracellular expression of Gal-3 in pancreatic cancer cell lines
under hypoxic microenvironment and starving condition. Methods: Two pancreatic cancer cell lines (PANC-1 and
MiaPaCa-2) were cultivated in DMEM supplemented with 10% FBS in a humidified atmosphere at 37⁰C with 5%
CO2. Cells were plated at 1x106 concentration and conditioned in hypoxic chambers under 1.5% pO2, in
presence and absence of serum. Then they were maintained at 37⁰C for 24 and 48 hours. Supernatants were
collected and Gal-3 levels were determinate by sandwich ELISA. Statistical analysis were performed using the
nonparametric test Mann-Whitney (GraphPad Prism® software version 6) and p values <0.05 were considered
significant. Results/Discussion: Gal-3 evaluation from supernatant of MiaPaCa-2 culture showed that its
extracellular levels were significantly elevated in simultaneous oxygen and nutrient deprivation conditions after 24
hours (p <0,05). None oxygen or nutrient deprivation isolated were capable of change significantly Gal-3 levels.
No significant changes were found in 48 hours conditions. Gal-3 in PANC-1 showed similar results. In 24 hours,
levels were increased in associated hypoxic and deprived nutrients condition (p<0.01) when compared to control
and isolated oxygen deprivation. Moreover, Gal-3 levels decreased in hypoxic conditions within 48 hours.
Conclusions: Simultaneous hypoxia and starving conditions lead to Gal-3 extracellular secretion, which does not
occur for longer time.
Financial Support
Fundação de Amparo à Ciência e Tecnologia do Estado de Pernambuco- FACEPE and Instituto Nacional de Ciência e Tecnologia para Inovação FarmacêuticaINCT-IF.
139
EVALUATION OF CELLULAR DISRUPTION AND ANTIOXIDANT ACTIVITY IN
DIFFERENT FRACTIONS OF Lactobacillus acidophilus
Luciana Oliveira de Fariña
Professor - Paraná West State University
Suzana Bender
Graduate Course - Pharmaceutical Sciences - Paraná West State University
Helder Lopes Vasconcelos
Professor - Pharmaceutical Sciences - Paraná West State University
Ana Carolina Costa
Undergraduate Course - Pharmacy - Paraná West State University
Introduction: Numerous investigations show that the lactic acid bacteria, have antioxidant systems,
especially Lactobacillus acidophilus, both in its intact cell lysate as in the extract of cells. Aim: The aim of this
study was to evaluate the efficiency of cell lysis to obtain the lysate extract cells and its antioxidant activity as
well as the intact cell, obtained from Lactobacillus acidophilus cultures, isolated from a strain originating from the
pharmaceutical industry. Material and Methods: Cell lysis was performed using sonication and centrifugation
processes recommended in technical literature and its efficiency was evaluated by poor plating of the samples
tested. The antioxidant activity was measured using the method of reducing free radical 2,2-diphenyl-1-picrylhidrazila (DPPH) with different concentrations of the sample. The result was compared to the positive BHT as a
standard. Results and Discussion: The results indicated that there was no lysis in cells culture evaluated and the
processes used only separated the colonies. It was noted that antioxidant activity increased in DPPH test,
proportionally to the concentration of the sample and was greater in sample obtained by sonication and
centrifugation (49.68%) than in intact cells (32.95%) and BHT standard at a concentration of 75μg / ml (42%).
Conclusion: Absence of cell lysis could be explained considering the thickness of Lactobacillus acidophilus cell
wall, which may have hindered the action of ultrasonic forces. Sonication separated the colonies, increasing the
number of intact cells present in the sample and the antioxidant activity. This activity can be attributed to the
exopolysaccharide present in their cell wall, composed of polymers with different physical properties able to
reduce DPPH, especially when more cells are present (sonicated sample).
Financial Support
CAPES; FUNDAÇÃO ARAUCÁRIA; UNIOESTE
Acknowledgments
To the financial agencies that support this research.
Ethical approval
Non aplicable
140
IN VITROEVALUATION OF LPSF/CR35 AS A PERSPECTIVE FOR FIBROSIS
TREATMENT
MARIA CLARA PINHEIRO DUARTE SAMPAIO
ANDERSON RODRIGUES DE ALMEIDA
MAIRA GALDINO DA ROCHA PITTA
MOACYR JESUS BARRETO DE MELO RÊGO
IVAN DA ROCHA PITTA
MARINA GALDINO DA ROCHA PITTA
MARIA DO CARMO ALVES LIMA
ANDREA TAVARES DANTAS
MICHELLY CRISTINA PEREIRA
It has been demonstrated that IL-6 induces extracellular matrix protein synthesis by fibroblasts in
fibrosis pathogenesis. Strategies to block the IL-6 response resulted in a significant reduction of pro-fibrotic
molecules in cultured fibroblasts. Some studies suggest that thiazolidinederivatives have antifibroticeffect. Our
study aims to evaluate the antifibrotic potential of a novel thiazolidine derivative. Fibroblasts cell line HFF-1 was
maintainedaccording to the datasheet provided by ATCC and plated for cytotoxicity by MTT assay. Cells were
plated at a concentration of 5x105 and treated with LPSF/CR35 at doses of 10 and 100 DM and with GW9662, an
antagonist of PPARy (10 DM). Non-treated cells,cells treated with Methylprednisolone at concentrations of 10 and
100 DM or Rosiglitazone at a concentration of 10 DM were used as controls. After 24 hours of incubation,
supernatants were collected and used to evaluate IL-6 levels by sandwich ELISA. Data were analyzed by
Wilcoxon test, with differences considered significant at p < 0.05.Derivative LPSF/CR35 showed no cytotoxicity
in HFF1 human fibroblasts. LPSF/CR35 significantly inhibited IL-6 production at doses of 10DM(p=0.043) and
100DM (p=0.025). IL-6 has an important association with several fibrotic diseases. Thus, molecules that inhibit IL6 action could represent an interesting alternative on fibrotic diseases treatment. The effects of LPSF/CR35 on IL6 production was not reverted by treatment with GW9662. Although thiazolidine derivatives are known to act
through PPARy, our results suggest that LPSF/CR35 acts independently of PPAR gamma, since its effect was
not affected by GW9662, an antagonist of PPAR gamma.Thiazolidine derivative LPSF/CR35 showed low toxicity
in HFF1 human fibroblasts and was able to inhibit significantly the production of IL-6 in these cells. Our results
also suggest that LPSF/CR35 acts independently of PPARy.
Financial Support
Foundation in Support of Science and Technology in the State of Pernambuco- FACEPE andNational Institute of Science and Technology for Pharmace utical
Innovation- INCT_if.
141
PLEIOTROPIC EFFECTS OF SIMVASTATIN WITH HUMAN DENTAL PULP CELLS
Maria Luísa de Alencar e Silva Leite
Department of Dental Materials and Prosthodontics, Araraquara School of Dentistry, UNESP, Araraquara, SP, Brazil
Diana Gabriela Soares
Fernanda Gonçalves Basso
Josimeri Hebling
Department of Orthodontics and Pediatric Dentistry, Araraquara School of Dentistry/UNESP, Araraquara, SP, Brazil.
Carlos Alberto de Souza Costa
Introduction. Simvastatin has been shown to feature anabolic effects with bone cells, increasing the
deposition of mineralized tissue. However, the pleiotropic effect of this substance on the regenerative potential of
dental pulp cells is not completely known. Therefore, the objective of this study was to assess the bioactivity of
simvastatin (SV) applied on human dental pulp cells (HDPCs) in vitro. Methods. The HDPCs were seeded on 96well plates at 80% confluence and exposed to the following treatments: NC - negative control (culture medium);
PC - positive control (10 ng/mL TGFβ1); SV1 1 µM/L SV; SV2 0.1 µM/L SV; and SV3 - 0.01 µM/L SV. Cell
viability (alamar blue), ALP activity (colorimetric assay) and mineralized matrix deposition (alizarin red) were
quantified after 1, 7, 14 and 21-day periods of incubation of cells in contact with the substances. Cell migration
was assessed up to 48 hours (wound healing/ transwell devices). Data was subjected to ANOVA and Tukey's
test; α=0,05. Results. SV1 decreased significantly the HDPCs viability at all time-points as well as reduced ALP
activity and mineralized matrix deposition. Significant lower percentage of cell viability than NC was detected for
SV2 and SV3 at 14 and 21-day periods. However, SV3 group featured significant higher ALP activity than NC at
the 7- and 14-day time-points. Intense mineralized matrix deposition was detected at 21 days for SV2 and SV3
groups related with NC and PC, with SV3 featuring significantly higher values than SV2. Wound healing assay
demonstrated that all SV concentrations reduced significantly the wound area compared to NC and PC, with the
best results being found for SV3. This concentration also induced intense active cell migration (transwell) after 4
and 24 hours in comparison to control groups. Conclusion. Simvastatin at low concentration induced dental pulp
cells migration and differentiation, culminating with deposition of high amounts of mineralized matrix.
Financial Support
CNPq grants # 303599/20146; FAPESP grants # 2014/235200 and # 2015/156357.
Ethical approval
Research Ethics Committee of the Araraquara Dental School/UNESP, São Paulo, Brazil - Proc. No 30939314.5.0000.5416
142
ASSESSMENT ACTIVITY OF CYTOTOXIC TIAZACRIDINES COUMPOUNDS IN PBMC´S
AND LEUKEMIA T-CELLS: IN VITRO STUDY
Mário Sérgio de Souza Albuquerque
Mário Sérgio de Souza Albuquerque
Artur Felipe Santos Barbosa
Pablo Ramon Gualberto Cardoso
Thércia Mayara Oliveira Feitoza
Ivan da Rocha Pitta
Introduction: Cancer is a term used to characterize multifactorial genetic changes that lead to
uncontrolled growth of cells and formation of malignant tumors. These cells can become invasive to tissue and
organs, spreading to other parts of the body. As a result of increased morbidity and mortality, cancer is a chronic
disease of social relevance. New clinical and pharmacological interventions are developed for prevention,
diagnosis and treatment of cancer. For example, tiazacridines compounds synthesized from the scaffold
tiazolidines and acridínicos cores, are potential candidates for anti-tumor drugs. Methods: The MTT assay was
used as is a colorimetric analysis based on conversion mitochondrial enzymes present only in metabolically
active cells. The Thiazacridine derivatives used in this experiment were: AA3, AA4, AA6, AA7, AA16 and AA19
The tiazacridines derivatives were tested at 1, 50 and 100 DM. All experiments were performed in triplicate and
viability calculation against DMSO. Results and Discussion: The cytotoxicity of the coumpounds was tested on
peripheral blood mononuclear cells and the compounds tested did not affect their cell viability. When we analyzed
the leukemic cells (Jurkat line), the AA-4 and AA-6 compounds showed reduced viability of tumor cells, in a
concentration of 50 µM, resulting in decreased cell on the order of 87.71% and 45.5%, respectively, compared to
untreated cells. The remaining compounds no showed reduction, in cell viability, above 25%, when compared to
cells that don't receiving treatment. Conclusion: The initial results demonstrate that, among the tiazacridinic
derivatives tested, the AA-4 and AA-6 compounds exhibited biological activity against tumor cell line (Jurkat),
and, they don't showed cytotoxic effect on the PBMC, at the same concentrations tested, becoming candidates
to the next assays.
Financial Support
FACEPE, INCT_if, CNPq
Acknowledgments
FACEPE, INCT_if, CNPq, UFPE, LINAT, NUPIT-SG, LPSF
143
ANTIBACTERIAL ACTIVITY OF BETULINIC AND URSOLIC ACID DERIVATIVES
AGAINST BIOFILM-FORMING BACTERIA
Muriel Primon-Barros
Faculty of Pharmacy, Federal University of Rio Grande do Sul, Porto Alegre, Rio Grande do Sul, Brazil
Gloria N.S. da Silva
Grace Gosmann
Simone C.B. Gnoatto
Alexandre J. Macedo
Faculty of Pharmacy, Federal University of Rio Grande do Sul, Porto Alegre, Rio Grande do Sul, Brazil; Centro de Biotecnologia, Federal
University of Rio Grande do Sul, Porto Alegre, Rio Grande do Sul, Brazil
About 80% of human infections are associated with bacterial biofilms, mainly those related to medical
implants, such as catheters, implants and prostheses, being a constant cause of nosocomial infection. Among
the strategies to combat biofilm formation on medical devices there is the search for new molecules with
antimicrobial and/or antibiofilm action. The triterpenes, as betulinic and ursolic acid, isolated from Platanus
acerifolia and Malus domestica, respectively, and derivatives can be potential active candidates against bacterial
biofilms. The aims of this study were to evaluate the antimicrobial and antibiofilm activities of betulinic and ursolic
acid and derivatives, and cellular toxicity. Experiments were performed using ATCC strains of E. fecalis, S.
aureus and S.epidermidis which were incubated with betulinic and ursolic acid and 20 derivatives solutions in
three different concentrations (100, 25 e 5 µM) for 24 h at 37 ºC and then antimicrobial and antibiofilm activities
evaluated and the crystal violet technique, respectively. The toxicity against VERO cells was test using the MTT
to evaluate the cell proliferation after 24 h in contact with the molecules solution. Among the 22 different
molecules tested the derivative GLB3 from betulinic acid and the derivative GLU3 from ursolic acid, that possess
the same structural modification showed against all gram-positive strains the most effective activity in
concentration of 100 µM. The MTT cell proliferation assay to test the toxicity of these molecules showed that
GLB3 and GLU3 inhibited 46 and 41% of cell proliferation, respectively. The results obtained showed that GLB3
and GLU3 from betulinic and ursolic acid have potential antimicrobial and antibiofilm against the Gram-positive
strains E. fecalis, S. aureus and S. epidermidis. More studies are needed to verify the potential application of
these molecules in the coating of surfaces of medical devices and to enhance aspects of toxicity.
Financial Support
CNPq and CAPES
Acknowledgments
This work was supported by grants and financial support from Brazilian Agencies CNPq and CAPES (Rede de Pesquisa e Formação em Biofuncionalização de
Superfícies ¿ Rede NANOBIOTEC-Brazil).
144
N-TERMINAL PRO-BRAIN NATRIURETIC PEPTIDE (NT-proBNP) LEVEL PRIOR THE
TREATMENT FOR HEAD AND NECK CANCER
Nazare Souza Bissoli
Department of Physiological Sciences, Federal University of Espírito Santo, Vitória-ES, Brazil
Karine Gadioli Oliveira
Daniely Souza de Nardi
Sandra Venturin von Zeidler
Nucleus of Biotechnology, Federal University of Espírito Santo, Vitória-ES, Brazil
Sonia Alves Gouvea
Introduction: The biomarker NT-proBNP is an important tool for diagnosis and prognosis and its level in
cancer patients are poorly studied, especially in head and neck cancer. Our study is aimed at evaluating whether
there is an increase in NT-proBNP levels in patients with unknown risk factors for elevated NT-proBNP. Methods:
Fifty three patients diagnosed with head and neck squamous cells carcinoma have been interviewed before
starting the anti-cancer treatment. Through the questionnaire, it was possible to evaluate gender, age, tumor site,
TNM-stage and self-reported of smoking/alcohol.Serum NT-proBNP levels were measured on the Cobas 6000
modular analyzer. The data were analyzed by the SPSS 17.0 software using Student t-test (p < 0.05). Results:
We found predominance of men (81.13%) and average age 58.7 ± 10. The primary site of tumor was usually oral
cavity, followed by the oropharynx, larynx and hypopharynx. Almost half of the cases (49%) were in advance
stage of cancer (stages III/IV) and there was no significant difference between the levels of NT-proBNP in
patients in early stage (89.03 pg/mL, range: 16.09-256.2 pg/mL) or advanced stage (98.44 pg/mL) of cancer.
Discussion and Conclusion: Although there was no significant difference in NT-proBNP levels between the
stages, the value of this marker remains higher than normal for head and neck cancer patients.As the main
treatment of these patients is chemoradiotherapy, this high value of NT-proBNP may bring further damage to the
patient leading them to worse prognosis. This result may assist the physicians in indicating a less aggressive
treatment for these patients whenever possible
Financial Support
CAPES, CNPq and FAPES
Ethical approval
Ethics Committee of the Espírito Santo Federal University nº 99.242/2012)
145
ANTIBACTERIAL AND MODIFYING-ANTIBIOTIC ACTIVITIES OF EXTRACTS AND
FRACTIONS OF Zornia reticulate Sm. AGAINST MULTIRESISTANT Streptococcus
RAFAELA CLEMENTINO HIBERNON
MANOELA FREIRE CLEMENTINO
Faculdade Maurício de Nassau (FMN)
ANA CLÁUDIA DANTAS DE MEDEIROS
RENALY IVYNA DE ARAÚJO RÊGO
Itavielly Layany França Feitosa
Thiago Pereira Chaves
Felipe Hugo Alencar Fernandes
Introduction:The emergence of multiresistant bacterial strains has led researchers to look for new
therapeutic alternatives, overcoming the resistance mechanisms. Amongst them, that is included the use of
natural products such as antibacterial or modifiers of bacterial resistance. Zornia reticulata Sm. (Fabaceae) is a
vegetal specie used in traditional medicine for treatment of various diseases, highlighting infections. This study
aimed to realize a phytochemical screening and evaluate extracts and fractions of Z.reticulata antibacterial and
modifiers of the resistance of genus Streptococcus bacterial to antimicrobial drugs. Methods: The crude ethanolic
extract (CEE) was obtained from the aerial parts of the plant, from which proceeded the fractionation by liquidliquid system partition with hexane system (FHex), chloroform (FChl) and ethyl acetate (FEthac).Phytochemical
tests were performed by addition of specific reagents and UV-VIS spectrophotometry. The minimum inhibitory
concentration (MIC) of natural products (PN) and antimicrobial drugs in the presence and absence of PN on
multiresistant strains of S. mutans and S. oralis was determined by microdilution.Results and discussion: It was
found steroids, tannins, flavonoids and saponins in CEE in all fractions. The FEtchac have higher concentrations
of polyphenols and flavonoids, while FHex showed higher concentration of tannins. The CEE, FChl and FHex
demonstrated significant antimicrobial activity against S. mutans (CIM < 1000 µg mL-1). When combined with
antimicrobial drugs insubnitory concentration (CIM/8), the PN significantly reduced the CIM of cefepime for both
strains.Conclusion: The results suggest that Z. reticulata presents PN with antibacterial activity and the ability to
modify the resistance of S. mutans and S. oralis to Cefepime, being a promising species for antibacterial therapy.
Financial Support
1Laboratory of Development and Assays of Drugs, Universidade Estadual da Paraíba, Campina Grande, PB, Brazil; 2Department of Biology, Universidade
Federal do Piauí, Bom Jesus, PI, Brazil3Faculty of Pharmaceutical Sciences, Universidade Estadual Paulista, Araraquara, SP, Brazil.
146
FREE-LIVING AMOEBA ISOLATED FROM COOLING TOWERS WATERS NATURALLY
INFECTED WITH Pseudomonas spp.
Scheila Soares Oliveira
Vlademir Vicente Cantarelli
Fundação Universidade Federal de Ciências da Saúde de Porto Alegre (UFCSPA)
Marilise Brittes Rott
Free-living amoebae (FLA) play an important role in the maintenance, evolution and act as carriers of
pathogens to humans and animals. FLA feed primarily on microorganisms and some have evolved to become
resistant to FLA. Pseudomonas spp. has intracellular growing capacity and will grow inside protozoans and
macrophages, which protects the bacteria against antibiotics and disinfectants. Respiratory vesicles released by
amoebae can contain hundreds of infective bacteria, and inhalation of even a few vesicles can lead to infection.
Artificial aquiferous systems, such as water supply facilities, especially heated reservoirs and cooling towers,
represent potential sources of environmental human infection. In this study, water from cooling towers was
collected and, after filtering, samples were cultivated on non-nutrient agar. DNA from cultivated amoeba were
extracted by a commercial system prior to submitting to PCR amplification with primers targeting Pseudomonas
spp. PCR reactions were performed and amplicons were separated by a 1.2% agarose gel. In total, 36 samples
were collected, from which 33 were positive by FLA. Pseudomonas spp was detected by PCR in 15 (45.4%) of
them. DNA sequence will be used to confirm these results and identify the microorganisms to the species level.
In summary, our preliminary work shows that FLA is very common in cooling towers waters and that
Pseudomonas spp constitutes an important intracellular organism found in these FLA. Further studies are under
way to determine the species involved and to detect other important intracellular candidate organisms associated
with human infections.
Financial Support
CAPES
147
EVALUATION OF in vitro ANTINEOPLASTIC ACTIVITY OF THIOSEMICARBAZONES
DERIVATIVES AGAINST K-562 CELLS
Thuanny Maryna dos Santos
Valéca de Cassia Mendonça da Costa
Ana Daura Travassos de Oliveira
Paulo André Teixeira de Moraes Gomes
Ana Cristina Lima Leite
Ivan da Rocha Pitta
Introduction: Cancer is characterized by dysregulated cell growth, result of alterations of genes involved
on cellular proliferation, migration and differentiation. Nowadays, cancer is the second cause of death in Brazil.
Currently available treatments are still associated to low selectivity, side effects and tumor resistance to
chemotherapy. Thiosemicarbazone derivatives has been described as potential anticancer agents. The purpose of
this work was to evaluate the anticancer activity in vitro of ten aryl thiosemicarbazone derivatives (LpQM/DTs) on
K-562 cells (chronic myelogenous leukemia). Methods: Cytotoxicity activity of LpQM/Dt-04, 08, 10, 12, 14, 22,
28, 32, 38 and 48 were performed using MTT method. Initially the toxicity in healthy fibroblasts was evaluated at
10, 50 and 100 µM. After that, it was performed cytotoxicity assays in K-562 cells at 1, 5, 10, 25 and 50 µM.
Evaluation of apoptosis inducing was performed by DNA electrophoresis after exposure at 25 µM of compounds.
Based on the results obtained, it was determinate the concentration of drug required for 50% inhibition of cell
viability (IC50) and selectivity index (SI). Results and discussion: All compounds are not toxic to healthy
fibroblasts in all doses tested. Cytotoxicity assays reveal that derivatives LpQM/Dt-04, 08, 10, 12, 14, 22, 28, 32,
38 and 48 shows antineoplastic activity in K-562 cells (IC50 = 11.40, 13.39, 14.05, 14.94, 9.17, 13.87, 11.32,
15.43, 17.29 and 14.80 µM respectively; SD < 10%). All selectivity indexes were higher than 3 and it is desirable
to have a high selectivity index, giving a maximum antitumor activity with minimal cell toxicity. Among them,
LpQM/Dt-08, 28 and 48 exhibited DNA fragmentation in electrophoresis gel, suggesting apoptosis induction.
Conclusion: Thiosemicarbazone derivatives display antineoplastic activity in K-562 cells, with high selectivity
indexes. Furthermore, LpQM/Dt-08, 28 and 48 seem to induce apoptosis.
Financial Support
CNpQ, INCT_if ,
148
149
SYNTHESIS OF NOVEL CYSTEINE PROTEASE INHIBITORS BY MOLECULAR
SIMPLIFICATION
Angélica Rocha Joaquim
Elany Barbosa da Silva
Isadora Serraglio Fortes
Débora Assumpção Rocha
Rafaela Salgado Ferreira
Saulo Fernandes de Andrade
Introduction: Chagas disease and the african trypanosomiasis are a problem of public health. The
cysteine proteases class is a promising therapeutic target for the development of new antitrypanosomal drugs.
Cruzain and rhodesain are the major cysteine proteases from T. cruzi and T. brucei, respectively, and these
enzymes present homology. In a High Throughput Screening campaign with a library of almost 200 thousand
compounds, it was identified a peptidomimetic chiral inhibitor bearing two stereocenters that was commercially
available in unknown ratio. The synthesis of the four stereoisomers of this is complex, thus in the present work it
is proposed the synthesis of simpler analogues of this lead compound, in addition to the evalutation of its activity.
Methods: Treatment of 2-furoic acid with L or D-valine methyl ester hydrochloride in presence of EDC, DMAP and
DIPEA gave S or R-amide, respectively. After the formation of the amide bond, the methyl ester of amides was
hydrolyzed in basic conditions and these were coupled to different amines to provide eight novel compounds. This
series was evaluated against cruzain and rodhesain using reported protocols by our group. Results and
discussion: By comparing the activities of the analogues synthesized, it became possible to establish a likely
structure-activity relationship. Regarding stereochemistry, in general, the most potent compounds possess R
configuration. The homologation of the amine chain or addition of a second ring resulted in an increase in activity.
Furthermore, the benzothiazole derivative showed relevant activity. Conclusion: Since the benzothiazole
derivative is considerably simpler than the lead compound and it can be synthesized in only three steps, it is
concluded that it was possible to identify a new lead compound by molecular simplification, which can be further
modified to obtain a new candidate for pharmaceutical drug.
Financial Support
CAPES, CNPq, FAPERGS.
150
SYNTHESIS AND EVALUATION OF NEW N-SUBSTITUTED-1,2-BENZISOXAZOLIN-3ONES AS ANTIFUNGAL DERIVATIVES
Antonio Maciel Fregnan
Universidade Federal de Alfenas (UNIFAL-MG)
Antonio Ávila Fernandes Junior
Josidel Conceição Oliver
Naiara Chaves Silva
Amanda Latércia Tranches Dias
Luiz Felipe Leomil Coelho
Thiago Belarmino de Souza
Danielle Ferreira Dias
Diogo Teixeira Carvalho
Introduction: Fungal infections caused by resistant microorganisms are constantly increasing and
severe side effects limit chemotherapy following. The search for new antifungal drugs is an urgent issue and
research on new structural scaffolds may lead to new substances with innovative mechanisms of action. The
enzyme H+-ATPase is an vital transport protein in the membranes of yeasts and some authors have shown that
eugenol, an important natural bioactive product, is able to inhibit its catalytic action and lead yeast cell to death.
Moreover, some benzisoxazolinones also show inhibitory action on this target. In view of this, our aim was to
synthesize a series of new N-substituted benzisoxazolinones designed from eugenol and analogues and to
evaluate them as antifungal candidates against Candida species. Methods: The desired benzisoxazolinones were
obtained in a six-step concise route from eugenol in a sequence that consisted of formylation, carboxylic acid
synthesis, phenol esterification, hydroxamic acid formation and Mitsunobu cyclization. All the substances were
characterized by melting point analyses, mass, IR and NMR spectroscopies. The anti-Candida sp. and cytotoxic
activities of final products and eugenol were checked in vitro by standard methods. Results and discussion: The
tests revealed that most of the synthesized compounds have higher antifungal activity than eugenol. N-phenyl
substituted benzisoxazolinones were the most promising ones, with IC50 values sometimes comparable to that of
standard drug fluconazole against C. krusei and C. glabrata and with good selectivity indexes. However, other
derivatives showed to be too cytotoxic to be considered in further studies of antifungal molecules. Conclusions:
We found that eugenol-based benzisoxazolinones are interesting derivatives to be optimized as antifungal drug
candidates and the next step is to evaluate their action on the target enzyme.
Financial Support
FAPEMIG (APQ-01209-13) and CAPES (scholarship, process 1416533)
151
In vitro and in vivo anti-melanoma effects of betulinic acid derivatives
Artur Stramari de Vargas
Elenilson Figueiredo da Silva
Jheini Lis Antunes Fernandes
Pontifícia Universidade Católica do Rio Grande do Sul (PUCRS)
Krist Helen Antunes Fernandes
Laura Trevisan Corrêa
Cristiane Bernardes de Oliveira
Grace Gosmann
Ana Paula Duarte de Souza
Rafael Fernades Zanin
Simone C.B. Gnoatto
Melanoma is one of the most important fatal skin diseases nowadays. Is has a high potential of
metastasize and there are a small number of pharmacological treatments available for it. An important triterpenoid
natural product, betulinic acid, exhibits several anticancer properties against a variety of cancer cell lines. In this
work, based on literature data, we selected the position C-3 of the betulinic acid, in order to exploit the
contribution of changes in this position against, melanoma cell line. Betulinic acid was extracted from Platanus
acerifolia. The semi-synthesis reactions were performed by substitutions in the C3-OH position by oxidation
reaction, oxime formation and reduction to amine. Three new derivatives were synthesized and tested against
melanoma (B16F10), VERO and HEP2 cells for 24h and the cell viability was assessed via MTT assay. A death
cell assay using FITC-conjugated annexin-V and propidium iodide was performed. Also, the antimetastatic effects
of BA derivatives in C57BL/6 mice treated with B16F10 melanoma cells were assessed. The oxime derivative
showed to be the most active compound. It caused a decrease of 96,8% of the melanoma cell viability after 24h
with a IC50 of 3,2μM and it was capable of really significant decrease of the tumor growth at 1.5 mg/Kg
concentration. So, the derivatives have shown to possess important potential for melanoma treatment and are
possible candidates to becoming new drugs.
Financial Support
CNPq/Brazil and master scholarship from CAPES/Brazil.
Acknowledgments
PPGCF/Faculty of Pharmacy, UFRGS.
Ethical approval
Consent of Ethics Committee on Animal Research (CEUA-PUCRS) (protocol 14/00394).
152
ALLYLIC THIOCYANATES AS A BROAD-SPECTRUM ANTIFUNGAL AGENTS
Daiane Flores Dalla Lana
Gabriela Machado
Bruna Pippi
Vanessa Bergamo
Bruna Batista
Letícia Danielli
Saulo Andrade
Marcus Sá
Universidade Federal de Santa Catarina (UFSC)
Gustavo Silveira
Alexandre Fuentefria
Fungal diseases are a global public health problem. The treatment of the fungal infections is often
considered long, expensive, and limited in terms of therapeutic options. The development of new alternatives for
the treatment of mycoses is extremely relevant. Herein, we present the screening of a series of allylic
thiocyanates against dermatophytes and yeasts including multidrug-resistant clinical isolates. Allylic thiocyanates
bearing halogenated aryl groups were readily obtained from the Morita-Baylis-Hillman adducts followed by bromide
nucleophilic substitution. The Minimum Inhibitory Concentration (MIC) was determined by microdilution method
following the CLSI guidelines. Especially chloro and bromine derivatives (four compounds) exhibited moderate-togood action (MIC ≤ 50 µg/mL), inhibiting in vitro growth of filamentous fungi and/or yeasts of Candida genus, as
well as multidrug-resistant species. These findings corroborate with preliminary antimicrobial results published by
our collaborators and other authors for this class of compounds. The synthesis of this collection of thiocyanate
allylic has been previously published. However, in synthetic terms there is no news, in other studies the collection
was tested against some other pathogenic fungi and some bacteria, with an excellent result. In addition, it was
also shown that the molecules are promising for the treatment of tuberculosis. In our study, these compounds
were evaluated for the first time against multidrug-resistant species, with action also for these clinical isolates of
difficult treatment. The antimicrobial activities of these allylic thiocyanates are very remarkable since some
compounds presented a broad spectrum of action. The straightforward preparation of these allylic thiocyanates
from inexpensive will allow further exploitation of this promising class of compounds as future leads for drug
development.
Financial Support
CAPES, CNPq, PPGCF, UFRGS
Acknowledgments
CAPES, CNPq, PPGCF, UFRGS
153
Interactions of insulin with calcium alginate from molecular dynamics simulations
Daniela Nadvorny
UFPE
Nanostructured systems formed by polymer such as calcium alginate, dextran sulfate and chitosan are
being studied as alternative drug delivery systems. These composites permit oral administration by protecting the
drug from intestinal enzymes and allowing absorption through the gastrointestinal tract. This is especially
interesting for insulin delivery, as the conventional method of subcutaneous administration is invasive and can
lead to side effects and dosage issues. Computational methods can assist in the understanding of the
interactions and mechanisms present in these systems, providing information that may be experimentally costly
or difficult to access. In this work, molecular dynamics methods were used to study nanoparticle systems formed
by insulin and calcium alginate. The objective of this work was to understand the interactions between the insulin
and calcium alginate. Calcium alginate oligomers in complex with insulin were simulated using the AMBER
packpage with ff14SB and GLYCAM force fields. Parameters for the guluronic acid in alginate were obtained by
modification of the 0GB and 4GB monomers from GLYCAM. The insulin structure used was obtained from Protein
Data Bank (PDB:1MSO). We show the key residues involved in this interaction, and the dynamics of insulin in
the calcium alginate matrix. Understanding these interactions will assist in the development of new alginatebased, oral insulin delivery systems.
Acknowledgments
CAPES, CENAPAD-PE
154
MECHANISM AND SELECTIVITY OF CYP2E1 INHIBITION BY CHLORMETHIAZOLE:
POTENTIAL USE IN ACETAMINOPHEN OVERDOSE
DANILLO EWERTON DOS SANTOS OLIVEIRA
Pharmaceutical Science Department, Federal University of Pernambuco, Recife, PE, Brazil
ROBERT PELLETIER
Medicinal Chemistry Department, University of Washington, Seattle, WA, United States
KENT KUNZE
Medicinal Chemistry Department, University of Washington, Seattle, WA, United States
Adverse drug reactions (ADRs) are an appreciably harmful reaction, resulting from an intervention
related to the use of a medication. They can cause harm to the patient and are also responsible for increased in
medical cost. Many ADRs are a consequence of drug-drug interactions (DDIs) that promote alterations in drug
metabolism. In fact, evidence suggests that the majority of DDIs are mediated through the induction or inhibition
of Cytochrome P450 enzymes isoforms (P450). Therefore, understanding P450 mediated DDIs and their impact
on drug metabolism is critical in preventing ADRs in humans. Chloromethiazole (CMT) is a FDA acceptable
inhibitor of CYP2E1 for in vitro studies; however, little is known about its utility as probe inhibitor. We are
interested in determining the mechanism and selectivity of CYP2E1 inhibition by CMT; we also seek to
demonstrate its utility as an inhibitor of acetaminophen (APAP) bioactivation by CYP2E1, which is relevant to the
treatment of overdose. To understand CMT's inhibitory mechanism, our goal is to synthesize deuterated analogs
of CMT and to elucidate its mechanism of irreversible enzyme inactivation via thiazole bioactivation.
Replacement of selective hydrogen atom with deuterium should help to identify metabolic sites that are
responsible for inactivation due to changes in the inactivation rate arising from kinetic isotope effects at these
metabolic sites. Further, selectivity of inhibition will be assessed in vitro for irreversible and reversible inhibition of
major CYP isoforms by experiments with accepted probe substrate and LC/MS/MS analysis. If the data support
these hypotheses, the drug could be tested with cryopreserved human hepatocytes as a model system to assess
acetaminophen toxicity. The completion of this project will help characterize CMT as a selective CYP2E1 inhibitor
and as a potentially beneficial drug to treat CYP2E1-mediated acetaminophen toxicity.
Acknowledgments
Professor Dr. Kent Kunze; Professor Dr. Ricardo Brandão; Professor Dr. Dalci Brondani; Robert Pelletier; Ryan Seguin
155
ASYMMETRIC SYNTHESIS OF FRAGMENT C1.C6 OF COMPETITIVE CHIRAL
INHIBITOR OF CRUZAIN.
Débora Assumpção Rocha
Programa de Pós-Graduação em Ciências Farmacêuticas - Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil.
Isadora Serraglio Fortes
Faculdade de Farmácia - Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil
Angélica Rocha Joaquim
Programa de Pós-Graduação em Ciências Farmacêuticas - Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil;
Elany Barbosa Silva
Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas - Universidade Federal de Minas Gerais, Belo Horizonte, MG,
Brazil
Rafaela Salgado Ferreira
Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas - Universidade Federal de Minas Gerais, Belo Horizonte, MG,
Brazil
Programa de Pós-Graduação em Ciências Farmacêuticas - Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil;
Introduction: Chagas disease is a neglected tropical illness, whose etiological agent is the protozoan
Trypanosoma cruzi. One of the validated drug targets for the treatment of the disease is cruzain, the major
cysteine protease of T. cruzi which plays essential role in parasite survival. In a recent study using docking and
High-throughput screening (HTS), the chiral compound N-[1-[[[2-(2-furanyl)-2-(1-piperidinyl)ethyl]amino]carbonyl]-2methylpropyl]-2-furancarboxamide (1) was identified as a non-covalent inhibitor of cruzain. In the present work the
asymmetric synthesis of the fragment (R)-2-(2-furanyl)-2-(1-piperidinyl)-1-azidoethane, which is the key
intermediate in the synthesis of the chiral cruzain inhibitor 1, is reported. Methods: The fragment was synthesized
in several steps starting from furfural and using commercially available reagents. Compounds were characterized
by its IR and NMR spectra. Results and Discussion: Initially, furfural was converted to the furanylpropenoic acid
by Doebner reaction. This acid was decarboxylated in reaction with Cu2O providing vinylfuran that was submitted
to asymmetric Sharpless dihydroxylation conditions to give the S enantiomer glycol. Protection of the hydroxyl
group using dimethyl carbonate followed by reaction with sodium azide provided the azido alcohol derivative
stereospecifically with inversion of configuration. The reduction of the azido group with H2 in the presence of
Lyndlar catalyst afforded the known amino alcohol that was treated with 1,5-dibromopentane to give the piperidinyl
derivative. Finally, the hydroxyl group was converted into the azido group by Mitsunobu conditions, giving the
desired fragment. Conclusion: The asymmetric synthesis of azidoethane fragment was achieved with moderate to
good yields. This compound is an important intermediate and its preparation is a limiting step in cruzain inhibitor
synthesis.
Financial Support
FAPERGS, CAPES, CNPq.
156
SYNTHESIS AND CYTOTOXICITY ACTIVITY OF LAPACHOL GLYCOSYL TRIAZOLES
Flaviano Melo Ottoni
Departamento de Produtos Farmacêuticos, Faculdade de Farmácia, Universidade Federal de Minas Gerais
Lucas Lopardi Franco
Departamento Química, Instituto de Ciências Exatas, Universidade Federal de Minas Gerais
Lucas Bonfim Marques
Departamento de Fisiologia e Biofísica, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais
Elaine Maria de Souza Fagundes
Departamento de Fisiologia e Biofísica, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais
Ricardo José Alves
Departamento de Produtos Farmacêuticos, Faculdade de Farmácia, Universidade Federal de Minas Gerais
Introduction: Lapachol is a naphthoquinone that can be isolated from plants of several species, like
Handroanthus impetiginosus (Purple ipê) and has been shown to be a potential antitumor agent. However, high
enough blood levels were not achieved to show the therapeutic expected effect, which preclude its clinical
development as medicine. Due to its antitumor activity, it is a good lead for the design of derivatives with
improved properties. We designed glycosyl triazoles derived from lapachol as potential antineoplastic agents.
Methods: The derivatives were synthesized from "click", reaction of peracetylated and deacetylated carbohydrate
azides (D-glucose, D-galactose, D-N-acetylglucosamine and L-fucose) with O-propargyl lapachol. The obtained
derivatives were evaluated in vitro against six tumor cell lines (Jurkat, HL60, MCF-7, MDA, THP-1 and HCT) and
normal cells (VERO). Results and Discussion: The derivatives were obtained in yields ranging from 49 to 84%
and showed general low cytotoxicity in normal cells. Almost all were active against at least one tumor cell line.
Derivatives of D-galactose and L-fucose were active against all strains, but were cytotoxic to normal cells. The
peracetylated derivative of D-glucose showed the lowest IC50 among all tested derivatives (IC50 = 11.6 µM
against HL60), as well as being the most active against THP-1 (IC50 = 20.8 µM). Among deacetylated lapachol
glycosyl triazoles, only D-galactose and L-fucose showed activity (D-galactose with IC50 = 53.1 µM against
HL60 and L-fucose with IC50 = 18.1 µM against HL60, IC50 = 43.2 µM against MCF-7 and IC50 = 22.4 µM
against HCT). Conclusions: a series of glycosyl triazoles derivatives of lapachol were prepared and evaluated
against six cancer cell lines. All the compounds were more active than lapachol and some displayed low
micromolar activity against at least one cancer cell line and showed general low citotoxicity against normal cells.
Financial Support
CAPES, FAPEMIG and CNPq.
157
5-AZA-2'-DEOXYCYTIDINE AS AN ANTIMETASTATIC AGENT IN BREAST CANCER
CELLS
Giseli Klassen
Federal University of Parana
Andressa Chequin
Mariana Busato Toledo
Graciele Cristine More Manica
Liliane Maria Bacaro Klassen
Yara de Oliveira Brandrão
Isis Venturi Biembengut
Edneia Amancio de Souza Ramos
Epigenetic changes have been increasingly recognized as a driving force in human breast cancer.
Aberrant methylation, for example, it's closely associated with tumorigenesis by increasing genomic instability or
by silencing tumor suppressor genes. These observations have led to recovery the interest in DNA methylation
inhibitors as antineoplastic agents in clinical trials. The prototypical 5-aza-2'-deoxycytidine (5-azadC) has recently
been approved by the US Food and Drug Administration as antitumor agent, because it incorporates into DNA
and traps DNA methyltransferase in the form of a covalent protein. As a result, cellular DNA methyltransferase is
rapidly depleted, and concomitantly genomic DNA is hypomethylated during continued DNA replication. The aim
of this study was investigate the effect of the antitumor agent 5-azadC in aggressive, mesenchymal and
metastatic breast cancer cell line (MDA-MB-231). Thereby both mock and 5-azadC treated cells were analyzed
for migration by wound healing assay, gene expression and methylation specific PCR (MSP) after bisulfite DNA
treatment. As a consequence of 5-azadC treatment, MDA-MB-231 showed less migratory activity than mock
cells. In a panel of 12 genes associated with breast cancer progression only CDH1 (E-cadherin adhesion protein)
was significantly activated after 5-azadC. The MSP analyzes showed that in the treated cells CDH1 promoter
gene was significantly hypomethylated. E-cadherin is closely related to the ability to hold together cancer cells
and consequently preventing metastasis. This is the first description showing that 5-azadC agent seems to be
promise in the treatment of breast cancer cells, by promoting DNA hypomethylation and consequent expression
of key genes capable of controlling the cells migration and metastasis. Financial Support: CNPq, Capes.
Financial Support
CNPq
Acknowledgments
CNPq, CAPES
158
ANTIFUNGAL AND CYTOTOXIC ACTIVITIES OF PIPLARTINE SYNTHETIC ANALOGUES
Guilherme Andrade Brancaglion
Antonio Maciel Fregnan
Naiara Chaves Silva
Stella Maria de Souza Morais
Josidel Conceição Oliver
Amanda Latércia Tranches Dias
Luiz Felipe Leomil Coelho
Danielle Ferreira Dias
Thiago Belarmino de Souza
Diogo Teixeira Carvalho
Introduction: Piplartine is an important piperamide found in Piper sp., which inhibits the phytopathogenic
Pyricularia grisea and shows activity comparable to nystatin and miconazole against Cladosporium
sphaerospermum and C. cladosporioides. Some piplartine-like piperamides showed promising activity against
Aspergillus flavus and Candida albicans. Following this, it is described here the synthesis of 16 piplartine
analogues and their evaluation against Candida sp. and cytotoxicity on a healthy cell line. Methods: Piplartine
analogues were obtained in few objective steps, starting by the acyl chlorides preparation from four cinnamic
acids (cinnamic, 4-methoxycinnamic, 3,4-dimethoxycinnamic and 3,4,5-trimethoxycinnamic acids), their reaction
with hydrazine hydrate and the reaction of the respective hydrazides with phthalic anhydride or benzoyl chloride to
afford the first group of derivatives. The reaction of the same acyl chlorides with morpholine or Nhydroxysuccinimide led to the other group of derivatives. These products and piplartine were evaluated in vitro
against Candida sp. by the microdilution method and fluconazole was employed as control of fungistatic action.
The cytotoxicity of these compounds were checked on healthy hamster kidney cell (BHK-21) by the MTT method.
Results and discussion: Among these piplartine analogues, morpholine 3,4,5-trimethoxylated derivative was
almost 4-fold more potent and about 5-fold less toxic than piplartine against C. krusei and showed a selectivity
index greater than 3. Furthermore, this derivative was 2-fold more effective than fluconazole against this strain.
This derivative also showed activity against C. tropicalis, while piplartine was inactive. Other analogues had
moderate activity against different fungal strains. Conclusions: We found that a piplartine analogue, namely (E)-1morpholino-3-(3,4,5-trimethoxyphenyl)prop-2-en-1-one, is an interesting derivative to be optimized as antifungal
drug candidate.
Financial Support
This work was supported by FAPEMIG (APQ-01209-13), CAPES, CNPq and FINEP.
159
EFFICIENT SYNTHESIS OF NEW THIAZOLIDINEDIONE GLYCOSIDES
Maralise Perigolo de Oliveira Carvalho
Flaviano Melo Ottoni
Mardonny Bruno de Oliveira Chagas
Maira da Rocha Galdino Pitta
Ricardo José Alves
Ivan da Rocha Pitta
Introduction: Thiazolidine-2,4-diones have been extensively studied due to their involvement in various
physiological processes such as cell proliferation, angiogenesis, inflammation, and glucose metabolism. Pitta et
al. prepared new benzylidene thiazolidine-2,4-diones with promising glucose lowering capability. However, these
compounds have low water solubility. As the attachment of hydrophilic moieties can improve this property, we
decided to investigate the synthesis of thiazolidinedione glycosides. Methods: thiazolidine-2,4-dione was reacted
with appropriately substituted 3-aryl-2-cyanoacrilate glycosides (method 1) or formylaryl glycosides (method 2) in
the presence of piperidine and ethanol at 95 ºC for 2 h to afford the corresponding thiazolidinedione glycosides.
Results and Discussion: in method 1, first the 3-aryl-2-cyanomethacrilate glycosides were prepared by reaction of
the appropriately substituted formyl- peracetylβDglycopyranoside with ethyl 2-cyanoacrilate in the presence of
piperidine and pyridine at 50 ºC for 2 h. The obtained glycosides were reacted with thiazolidine-2,4-dione to afford
the desired thiazolidenedione glycosides in 34% overall yield (2 steps). In method 2 we investigated the synthesis
of the thiazolidinedione glycosides by reacting the thiazolidine-2,4-dione directly with the formyl glycosides
employing the same conditions used in method 1. The title compounds were obtained in 45-64% yield. Method 2
has the advantage that the final compounds are obtained in one single step, in higher yields, simply and quickly,
by filtration, whereas method 1 requires two steps and product purification by column chromatography.
Conclusions: a series of thiazolidine-2,4-dione based aryl glycosides were successfully prepared by two methods,
opening the way for the evaluation of their physicochemical and biological properties. Financial Support: CNPq,
INCT-if.
Financial Support
CNPq, INCT-if.
Acknowledgments
INCT-if
160
SYNTHESIS AND CYTOTOXIC ACTIVITY OF NITROINDOLES AS BIOREDUCTIVE
ANTICANCER AGENTS
Marcela Silva Lopes
Jonas Pereira Ramos
Elaine Maria de Souza Fagundes
Renata Barbosa de Oliveira
Introduction: Hypoxia is a feature of most tumors and contributes for chemoresistance
and
radioresistance in cancer therapy. Bioreductive agents are prodrugs that are activated by reduction selectively
under hypoxic conditions being active only in tumor tissues. Nitro compounds have the potential to be
metabolized under hypoxia acting as bioreductive prodrugs. Based on this, we decide to investigate the cytotoxic
effect of nitroindoles against cancer cell lines under normoxia and model of hypoxia (3D culture) in vitro. Methods:
Nitroindoles were synthesized starting from acetone and phenylhydrazine, in 7 steps. These compounds were
assayed in vitro against three cancer cell lines (MDA-MB, MCF-7 and HCT) using 2D and 3D cell culture, at 50
µM. Compounds were also tested against normal cells (VERO) as parameter of toxicity. Results and Discussion:
Compounds were obtained in yields ranging from 16 to 89%. In general, nitroindoles exhibited an increased
activity in hypoxia model compared to normoxia against MCF-7 and HCT. However, they showed modest values
of percentage of inhibition. Among the 7 indoles evaluated, two exhibited activity under hypoxia at least 15-fold
greater than under normoxia against HCT, with inhibition values of 54% and 44%. The increase of activity in
hypoxic conditions may be due to bioreduction of nitro group, with formation of reactive radical species.
Specifically for these 2 compounds that bear a substituent in side chain at 3-position, an alkylating intermediate
can be formed after reduction, which explains the best results. Furthermore, compounds were not active against
normal cells indicating low toxicity. Conclusions: Two nitroindoles synthesized showed good selectivity profile to
hypoxic cells and low toxicity to normal cells being promising in the development of a new class of bioreductive
agents. Further studies are needed to find analogues with improved activity.
Financial Support
FAPEMIG, CAPES and CNPq
161
Marine Natural Products with Potential Anticholinesterase Activity: Molecular
Modeling Studies
NELILMA CORREIA ROMEIRO
UNIVERSIDADE FEDERAL DO RIO DE JANEIRO, CAMPUS MACAÉ PROFESSOR ALOISIO TEIXEIRA
KARINA GODARTH GONÇALVES
LIGIA MARIA DIAS DAUD
ANGELICA RIBEIRO SOARES
Title: Marine Natural Products with Potential Anticholinesterase Activity: Molecular Modeling Studies.
Authors: Godarth, K; Daud, LMD; Soares, AR; Romeiro, NC. Affiliation: PPG-ProdBio-Programa de PósGraduação em Produtos Bioativos e Biociências, Universidade Federal do Rio de janeiro, Campus Macaé
Professor Aloísio Teixeira, RJ, Brasil. Introduction: Alzheimer's Disease (AD) is a pathology related to age,
characterized by early symptoms like short-term memory loss, attention deficit disorder, mood swings and
behavioral problems. The causes are unknown but the amounts of acetylcholine, which is degraded by
acetylcholinesterase (AChE), are reduced, and AChE inhibitors are used to avoid this action. Objectives: This
project aims to investigate the inhibitory potential of the natural product elatol isolated from Laurencia seaweed
and derivatives by molecular docking with AChE as a basis for synthesis and pharmacological evaluation.
Methodology: Crystal structures of AChE were obtained from PDB. The reference ligand was retrieved from the
crystal and redocked. Spartan'08 was used to build and optimize Elatol and derivatives. Pharmacophoric groups
were added and modeled according to physiological pH. GOLD Suite version 4.1.2 was used to perform molecular
docking and Pymol v. 0.99 and Discovery Studio v. 3.1 were used to evaluate ligand-protein interactions. Finally,
ADMET descriptors were calculated with Marvin v. 6.2.1. Results and Discussion: This study suggested 28
candidates as AChE inhibitors. The best docking results showed relevant interactions, such as hydrogen bond
and pi-stacking, at two of the enzyme's active sites, suggesting a dual action which would relieve cognitive deficit
through the increase in neurotransmitter levels and prevention of Aß peptide aggregation. Conclusions: The
docking analysis suggested that the planned Elatol derivatives may be novel dual inhibitors of AChE. Financial
Support: FAPERJ; CNPq. Acknowledgements: GPNOA; LASSBio
Financial Support
FAPERJ, CNPq
Acknowledgments
GPNOA, LASSBio
Ethical approval
YES
162
NEW THIAZOLIDINEDIONES DERIVATIVES AND THEIR EFFECTS ON TH1 CYTOKINES
FROM PERIPHERAL BLOOD MONONUCLEAR CELLS OF PATIENTS WITH PSORIASIS
Mariana Modesto de Andrade Lima
Therapeutic Innovation Postgraduate Program, Laboratory of Immunomodulation and New Therapeutic Approaches (LINAT), Federal
University of Pernambuco, Recife, Brazil
Emerson de Andrade Lima
Clinics Hospital, Federal University of Pernambuco, Recife, Brazil.
Michelly Pereira Cristiny
Laboratory of Planning and Drug Synthesis (LPS), Center for Research in Therapeutic Suely Galdino (NUPIT-SG), Federal University of
Pernambuco
Claudia Diniz Lopes Marques
Ivan da Rocha Pitta
Laboratory of Planning and Drug Synthesis (LPS), Center for Research in Therapeutic Suely Galdino (NUPIT-SG), Federal University of
Pernambuco
Angela Luzia Branco Pinto Duarte
BACKGROUND: Psoriasis (Pso) is a chronic dermatitis that affects about 2-3% of the population. The
etiology and pathogenesis of Pso are still unclear, and has been treated as an inflammatory disease characterized
by Th1/Th17 imbalance. The Th1 pathway is known for the production of Interferonγ and TNF cytokines. These
cytokines act in keratinocytes proliferation which aggravates the psoriatic lesion. TNF is given as a crucial factor
on the onset of inflammatory diseases. IFNγ inhibits apoptosis of the keratinocytes and thus increases their
proliferation. We decided to study three new thiazolidinediones derivatives in PBMC from patients with Pso and
evaluating their potential to reduce Interferonγ and TNF levels. METHODS: PBMCs cultures stimulated or not
with PMA/Ionomycin were incubated in the presence of the new Thiazolidines derivate LPSF-SF33, -SF34 and SF35 compounds. Methylprednisolone (100µM) was the standard drug. After 48h at 37°C and 5% CO2, the
supernatants were used for Interferonγ and TNF evaluation by ELISA. The statistical test used was D'Agostino
omnibus normality test and Wilcoxon signed-rank test. RESULTS AND DISCUSSION: The compound LPSFSF33 was able to reduce Interferonγ at 50µM (P=0.0258) and TNF at 10µM (P=0.0353), 50µM (P=0,0052) and
100µM (P=0.0031). The LPSF-SF34 also reduced Interferonγ at 50µM (P=0.0182) and TNF levels at 50µM
(P=0.0012). The LPSF-SF35 was unable to reduce Interferonγ and TNF inflammatory cytokines. CONCLUSION:
The LPSF-SF33 and LPSF-SF34 compounds show promising results in decreased levels of Interferonγ and TNF
cytokines in Pso patients. The LPSF-35 needs more studies to show satisfactory results. ETHICAL APPROVAL:
Human Research-CEP-CCS-UFPE: 528/11 FUNDING SUPPORT: INCT_if, FACEPE, CNPq and CAPES
Financial Support
INCT_if, FACEPE, CNPq and CAPES
Acknowledgments
INCT_if, FACEPE, CNPq and CAPES
Ethical approval
Human Research-CEP-CCS-UFPE: 528/11
163
DETERMINATION OF ANTIFUNGAL ACTIVITY OF 8-HYDROXYQUINOLINE
DERIVATIVES ON PATHOGENIC FUNGAL SPECIES
Paula Reginatto
Bruna Pippi
Vanessa Zafaneli Bergamo
Gabriella da Rosa Monte Machado
Daiane Flores Dalla Lana
Lucas Lopardi Franco
Ricardo José Alves
Introduction: Since the late 60s, when antibacterial therapies were developed, there was a drastic
increase in fungal infections. Today, the resistance to antifungal agents is a major limitation for the treatment of
these infections.Thus, it is necessary to introduce new antifungals to combat persistent infections. In this
context, 8-hydroxyquinoline derivatives may have an important role in fungal therapy. The objective of this study
was to evaluate the anti-Candida and anti-dermatophyte activity 8-hydroxyquinoline derivatives: 8-hydroxy-5quinolinesulfonic acid (1), 8-hydroxy-7-iodo-5-quinolinesulfonic acid (2) and clioquinol (3) in order to gain insight
into the potency and the spectrum of activity of these compounds and to find novel antifungical hits to drug
design. Methods: For this, it checked the Minimum Inhibitory Concentration (MIC) of these compounds for
isolates of C. albicans, C. glabrata, C. krusei, C. parapsilosis, C. tropicalis, M. canis, M. gypseum, T.
mentagrophytes and T. rubrum. The MIC was determined by the broth microdilution method according M27-A3
and M38-A2 protocols (CLSI 2008). Results and Discussion: 8-hydroxyquinoline derivatives showed antifungal
activity against all isolates tested. The MIC values for dermatophytes were higher than for Candida sp.
Compound 3 was the most active for all species with MIC of 0.0625-2µg/mL, but there are many reports in the
literature on its toxicity. Compound 2 presents MIC values ranging 2-1024µg/mL and the MIC for the compound 1
ranged 1-512 µg/mL. The advantage of this latter substance in relation to other analogues is the absence of a
halogen substituent, which could decrease the toxicity. Conclusions: Finally, analogues of 8-hydroxyquinoline
showed satisfactory results of in vitro susceptibility testing, confirming its potential as a novel hits for antifungal
drug design. However, it is necessary more studies about mechanism of action and verification of toxicity.
Financial Support
CAPES, CNPq.
164
SYNTHESIS OF NOVEL THIAZACRIDINE DERIVATIVES AS ANTICANCER AGENTS
RENATA RODRIGUES DE CARVALHO
Michael Williams Leal
Moacyr Jesus de Melo
Ivan da Rocha Pitta
INTRODUCTION:Acridine and their derivatives represent a very interesting class of molecules due to
their broad spectrum of activity against bacteria, protozoa, malaria, cancer and HIV. They occur mainly through
molecular hybridization, which combines the pharmacophoric centers: thiazolidine acridine and, within the same
chemical entity obtaining more power, safety and efficacy. The mutagenic properties of these compounds depend
on the ability to intercalate with DNA bases pairs, leading to cell cycle arrest due to inhibition of the enzyme
topoisomerase I, II and apoptosis. METHOD:Novel synthetized thiazacridines derivatives were synthetized
through three reaction steps: n-alkylation reaction, reaction of condensation of Knoevenagel and Michael addition
reaction. They were evaluated in vitro as potent anticancer agents in different cancer cell lines. These cells were
subjected to anti-proliferative and apoptosis assays to elucidate the mechanism of cytotoxicity. RESULTS AND
DISCUSSION:Only the compound LPSF/AA-04 exhibited antiproliferative activity against acute T cell leukemia
with IC50 value of 30.14 ± 0.65µM. In cell death assays, LPSF/AA-04 derivative showed a significant increase in
the percentage of apoptosis compared with the untreated controls cells. In cell cycle analysis, compound
LPSF/AA-04 did not alter cell cycle in tumor cell evaluated. Therefore, these results suggest that apoptosis
induction was cytotoxicity mechanism of thiazacridine derivatives LPSF/AA-04. CONCLUSION: In conclusion,
the thiazacridine LPSF/AA-04 compound might be potential candidate for anticancer drug development, presenting
relevant biological results and good product based in synthesis technique.
Financial Support
UFPE, FACEPE, CNPq/INCT_if
Acknowledgments
UFPE, FACEPE, CNPq/INCT_if
Ethical approval
not needed
165
A COMBINED DOCKING-PHARMACOPHORE APPROACH FOR CYCLOOXYGENASE
INHIBITION PROFILE FROM ASTERACEAE FAMILY.
Ricardo Pereira Rodrigues
Faculty of Pharmaceutical Sciences, University of Sao Paulo (FCFRP-USP), Ribeirao Preto, SP, Brazil.
Fernando Batista da Costa
Faculty of Pharmaceutical Sciences, University of Sao Paulo (FCFRP-USP), Ribeirao Preto, SP, Brazil.
Cyclooxygenase (COX) metabolizes arachidonic acid into prostaglandin (PG)H2, the precursor for the
biosynthesis of various PGs, thromboxanes and prostacyclin. A low rate of gastrointestinal dysfunction caused by
a selective COX-1 inhibitor, SC-560, indicates that COX-1 selective inhibitors are still useful without causing
gastric damage. On the other hand, COX-2 selective inhibitors exhibit undesirable side effects, including
cardiovascular risk. Natural products are still a rich source to find novel structures and the anti-inflammatory
activity
from
Asteraceae
family
were
investigated
using
the
AsterDB
database
(http://www.asterbiochem.org/asterdb). Although COX-1 and COX-2 have distinct and preferable binding modes,
there is evidence that COX-1 inhibitors can also interact in the selective pocket proposed for COX-2 inhibitors
depending on the inhibitor's chemical structure. Analyzing 24 COX-1 PDB protein complexes, a docking protocol
in MOE software that includes a pharmacophore model as the first placement method to fit the ligand in the
binding site was performed and two pharmacophore models were developed to describe the two binding modes for
COX-1: the first, related to the indomethacin binding mode and the other one, describing the interaction in the
selective side pocket. Molecular interaction field were calculated: aromatic (energy -2.9 kcal.mol1); carbonil
(energy -3.7 kcal.mol-1) and hydroxyl (energy -7.5 kcal.mol-1). The combined docking-pharmacophore protocol
explores the binding mode of COX-1 inhibitors taking into account the 3D information of the two pharmacophore
models during the docking calculations to find new potential hits for COX-1 in the Asteraceae family.
Acknowledgments: Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP). Financial Support:
FAPESP (2013/26060-0).
Financial Support
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP): 2013/26060-0.
Acknowledgments
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP).
166
PM6D3 ENERGY EVALUATION OF INCLUSION COMPLEXES BETWEEN β
CYCLODEXTRIN AND 2HYDROXYPROPYLβCYCLODEXTRIN WITH β
CARYOPHYLLENE
SOFIA SANTOS DA SILVA
Federal University of Rio Grande do Norte
JONAS GABRIEL DE OLIVEIRA PINHEIRO
EUZÉBIO GUIMARÃES BARBOSA
ÁDLEY ANTONINI NEVES DE LIMA
Introduction: From Copaifera multijuga Hayne trees is extracted an oil resin known commonly as "óleo
de copaíba". It has βcaryophyllene (BCP) as major compound with anti-inflammatory, antitumoral and
antinociceptive proven activities. However, the compound hydrophobicity does not contribute to its bioavailability
for oral use. Cyclodextrins are a group of trunk-shaped oligosaccharides being widely used due to its well-defined
structure, low toxicity and capacity of forming inclusion complexes with hydrophobic molecules in its cavity. A
very useful method to study the interaction at atomic level is molecular modeling. Besides predicting possible
interactions, it is a powerful tool to give information such as interaction energy, geometry and stoichiometry
between a host and a guest molecule on molecular complex formation. The present work aimed to make a
theoretical complexation study between βcyclodextrin (BCD) and 2hydroxypropylβcyclodextrin (HPBCD) with
BCP in order to estimate the usefulness to improve the molecule solubility profile. Methods: BCP molecular
models were built and a conformational search was performed by molecular dynamics simulations. The two most
stable conformations were steered into the host molecules computing PM6-D3 semi-empirical
energy
minimizations every 1.0 Å steps. A potential energy plot was obtained to estimate the best route of entry as well
as the most stable complexes. Results and discussion: The lowest energy for the complex with BCP and BCD
was -20.93 kcal/mol and BCP with HPBCD was -45.65 kcal/mol. In terms of energetic values, the complexation
of the βCP in the HPBCD is easier to occur. Conclusion: The lowest interaction energy provided by BCP and
HPBCD complex indicates that, perhaps, this system will have a better performance in experimental studies.
Subsequently, experimental assays are going to be performed to support the in silico evidences of complexation.
167
STRUCTURAL AND STABILITY ANALYSIS OF LUNASIN PEPTIDE: A PROMISING
AGENT WITH CHEMOPREVENTIVE AND CHEMOTERAPEUTIC ACTIVITY AGAINST
CANCER
Stephanny Miranda Alves de Souza
Dario Eluan Kalume
Fundação Oswaldo Cruz
Luis Mauricio Trambaioli da Rocha e Lima
Theo Luiz Ferraz de Souza
Lunasin is a peptide discovered in soybean that has arisen as a promising chemopreventive and
chemotherapeutic agent against many cancer cell lines. In view of its possible use as a biopharmaceutical, a
better understanding of its structure and stability can allow a better understanding of its physiological action.
Here, we analyzed lunasin secondary and tertiary structures besides oligomerics states formation in simulated
physiological conditions, such as gastric and plasmatic pH. For this, we used circular dichroism (CD),
fluorescence spectroscopy, electrospray ionization-ion mobility spectrometry-mass spectrometry (ESI-IMS-MS),
high performance liquid chromatography (HPLC) and bioinformatics. Analyses of lunasin secondary structure by
CD has suggested that this peptide is majority disordered having a small content of βstrand that is more
stabilized in acid pH. CD experiments with TFE addiction suggested lunasin propensity to αhelix structure.
Denaturation process analysis suggested that lunasin has different conformations at pH 1.5 and 7.4.
Hydrophobicity scale of Kyte and Doolittle and IDPs predictors suggested that lunasin is an IDP. ESI-IMS-MS
and HLPC analysis indicated that lunasin is monomeric in solution, although ESI-IMS-MS has suggested
oligomeric states formation in low amount rising pH, which were not observed by HPLC. Light scattering analysis
suggested that lunasin has low tendency to aggregation that occured only in low intensity in its p.I. (pH 4.4).
BLASTP alignment has not suggested lunasin homology with human proteins and indicated similarity with seed
storage proteins. Therefore, our results suggests that lunasin is monomeric in solution having a majority
disordered structure with a βstrand content that suffer pH influence. Our results also indicate that lunasin is an
IDP with high structural flexibility with capacity to adopt different conformations what can explain its different
actions reported.
Financial Support
CNPq and FAPERJ
168
LPSF/AC-34 + LPSF/AC-129 THIAZACRIDINIC MIXTURE INDUCES CYTOTOXIC EFFECT
AND CAUSES POSSIBLE GENIC AND PROTEIN EXPRESSION MODULATION IN
LEUKEMIC CELLS
THIAGO UBIRATAN LINS E LINS
LINAT / NUPIT / UFPE
MICHELLY CRISTINY PEREIRA
MARDONNY BRUNO DE OLIVEIRA CHAGAS
KAMILA DE MELO VILAR
RICARDO OLÍMPIO DE MOURA
LPSF / UFPE
LPSF / UFPE
MARIA DO CARMO ALVES DE LIMA
LPSF / UFPE
IVAN DA ROCHA PITTA
LPSF / UFPE
Introduction: Searching for an antineoplastic compound that presents less cytotoxicity against normal
cells and more effectiveness against leukemic cells, we tested the LPSF/AC-34+LPSF/AC-129 thiazacridinic
mixture effect against hematopoietic tumor cell lineages HL-60 (human myeloid leukemia) and CCRF-CEM
(human lymphoid leukemia) and peripheral blood mononuclear cells (PBMC). Methods: Initially, the cytotoxicity
and selectivity were evaluated through MTT assay. After, possible changes in PPARγ, p21, BID, GADD153 and
RIP-3 gene expression caused by mixture compound was investigated through RT-PCR using GAPDH reference
gene as control. Additionally, it was elucidated pPTEN, NFκB p65, Bax, GADD 153 and PPARγ protein
expression in a 24h period through western blotting analysis using βactin as control. Values are showed as three
independent experiments mean in triplicate. Software Origin 8 was used to determine IC50 values. Unpaired
Student t-test (p<0.05) was performed for statistical analysis. Results and Discussion: The LPSF/AC34+LPSF/AC-129 mixture showed high selectivity to inhibit cell growth of HL-60 (IC50 = 8.05 μM) and CCRFCEM (IC50 = 14.27 μM) lineages when compared to PBMC. In HL-60, the genes GADD153 and PPAR presented
p values of p=0.0808 and p=0.2277, respectively. Both of genes had their expression compared to GAPDH
reference gene: 12.4701 fold and 4.8202 fold, respectively. In CCRF-CEM gene expression, the p value obtained
for SOD1 was p=0.0172 and 1.2995 fold expressed related to GAPDH. In HL-60 protein expression the p value
obtained for Bax and NFkB was p=0.9221 and p=0.3012, respectively. Conclusions: Considering the cytotoxicity
results, the mixture showed effective to inhibit the hematopoietic tumors proliferation without affecting normal
cells. But for other results, more tests will be assayed to elucidate the mixture role on apoptotic process induction
and genic and protein expression modulation.
Financial Support
Fundação de Amparo à Ciência do Estado de Pernambuco (FACEPE)
Acknowledgments
Instituto Nacional da Ciência e Tecnologia-Industria Farmacêutica (INCT-IF)
Ethical approval
CEP/CCS/UFPE 11006/2012
169
METABOLITE PRODUCED BY Fusarium oxysporum INHIBITS CAPSULE AND BIOFILM
FORMATION OF Cryptococcus spp.
William Lopes
Rafael Gomes Von Borowski
Simone Cristina Baggio Gnoatto
Marilene Henning Vainstein
INTRODUCTION: The use of medical devices continues to increase; yet their utilization are all often
complicated by infections with biofilm-forming microorganisms. Biofilms are communities of microorganisms
attached to a solid surface enclosed in an exopolymeric matrix. Cryptococcus spp. yeasts produce a
polysaccharide capsule and can form biofilms on medical devices. It was estimated 957.900 cryptococcal cases
and approximately 624.725 deaths in 2006. This work aims to evaluate the inhibition of C. neoformans and C.
gattii biofilms by using metabolites produced by filamentous fungi isolated from tree bark. METHODS: For
production of metabolites, 37 filamentous fungi were grown in two different liquid culture media for 21 days. The
supernatants were filtered with 0,22 μm membrane. The biofilm modulation assays were performed using a
microtiter plate model, microscopic examinations, and XTT reduction. RESULTS: Antibiofilm activity was found in
11 filamentous fungi supernatant. Sequencing of 18S rDNA was used for strain identification. To elucidated
antibiofilm mechanism, the supernatant produced by Fusarium oxysporum was selected. The minimum biofilm
inhibitory concentration (MBIC) was determined and the concentration used to evaluate the effect on the C. gatti
and C.neoformans capsules size. The results showed 100% capsule inhibition when Cryptococcal cells were
treated with MBIC concentration. The inhibition was confirmed by immunofluorescence employing an anti-GXM
antibody and scanning electron microscopy, supporting the relation between capsule inhibition and inhibition of
adhesion to surfaces. The isolation and characterization of bioactive compounds through a set of
spectrophotometric and chromatographic techniques are in progress. CONCLUSIONS: This fact suggests an
option for the prevention of fungal biofilms on medical devices. Our findings also demonstrate the potential of
filamentous fungi in the production of bioactive compounds against biofilms
Financial Support
CAPES, CNPQ
Acknowledgments
Laboratório de Fungos de Importância Médica e Biotecnológica and Centro de Biotecnologia da UFRGS
170
171
ANTI-TRYPANOSOMA CRUZI ACTIVITY OF ROOT AND SEEDS EXTRACTS FROM
LONCHOCARPUS CULTRATUS
ALINE GRIEBLER
Center for Medical and Pharmaceutical Sciences, State University of West Paraná - UNIOESTE, Cascavel, PR, Brazil.
ALINE ANTUNES MACIEL BORTOLUZZI
PATRICIA KAROLINE MATOS
RAFAEL ANDRADE MENOLLI
LÍVIA GODINHO TEMPONI
Center of Biological Sciences and Health, State University of West Paraná - UNIOESTE, Cascavel, PR, Brazil.
EDSON ANTONIO ALVES DA SILVA
Center of Exact Sciences and Technology, State University of West Paraná - UNIOESTE, Cascavel, PR, Brazil.
TEREZA CRISTINA MARINHO JORGE
Introduction: The American trypanosomiasis is caused by the protozoan Trypanosoma cruzi and
classified as a neglected tropical disease, although it has high rates of mortality and is endemic in 21 countries in
Latin America, with about 65 million people at risk of contracting the disease. The trypanosomiasis treatment
therapy is scarce, inefficient and numerous side effects. Taking into account these data, the objective of this
study was to investigate the anti-Trypanosoma cruzi activity of root and seeds extracts from the Lonchocarpus
cultratus. Methods: A voucher specimen of the plant was identified as L. cultratus (Vell.) A.M.G. Azevedo & H.
C. Lima and deposited in the herbarium of UNIOESTE (UNOP no. 20). Samples of roots and seeds were
collected, dried, grinded and subjected to exhaustive and successive extractions with hexane. After filtration and
evaporation of the solvent there was obtained the extract of the roots (LHR) and seeds (LHS). The antiTrypanosoma activity test was conducted by employing forms epimastigotes of T. cruzi, Y strain, grown in Liver
Infusion Tryptose (LIT) and the extracts evaluated front and Benznidazole control at concentrations of 1, 10, 15,
50, 100, 150 and 175 μg.mL1. The parasite growth was observed after 72 hours of incubation, parasite by direct
counting in a Neubauer chamber and the assessment of anti-T. cruzi activity, by concentration of the extract able
to inhibit the growth of protozoa 50% (IC50). Results and Discussion: The values of IC50 of LHR (13.94 µg.mL-1)
and LHS (42.58 µg.mL-1) were superior to the Benznidazole (2.97 µg.mL-1), positive control of the test and drug
used in the treatment of trypanosomiasis. These results indicated that the extracts have anti-T. cruzi activity and
should be studied because despite the Benznidazol medicine have lower IC50 value, it produces many side
effects. Conclusion: LHR and LHS extracts have anti-Trypanosoma cruzi activity.
Financial Support
Foundation Araucaria and Higher Education Personnel Improvement Coordination (CAPES)
172
TOXICITY OF Urera baccifera (URTICACEAE) LEAVES IN DNA DOUBLE-STRAND
BREAKS TEST
Faculty of Pharmacy, Integrated Regional University of High Uruguay and Missions, Santiago Campus, Santiago, Rio Grande do Sul, Brazil.
And Pharmaceutical Sciences Graduate Program, Federal University of Santa Maria, Santa Maria, Rio Grande do Sul, Brazil.
Fernanda Barbisan
Pharmacology Graduate Program, Federal University of Santa Maria, Santa Maria, Rio Grande do Sul, Brazil
Ivana Beatrice Mânica da Cruz
Pharmacology Graduate Program, Federal University of Santa Maria, Santa Maria, Rio Grande do Sul, Brazil
Margareth Linde Athayde
Pharmaceutical Sciences Graduate Program, Federal University of Santa Maria, Santa Maria, Rio Grande do Sul, Brazil.
Sydney Hartz Alves
Pharmaceutical Sciences Graduate Program, Federal University of Santa Maria, Santa Maria, Rio Grande do Sul, Brazil.
Introduction: Urera baccifera is a shrub known as red nettle or stinging nettle, due the presence of
stinging hairs that cause irritating effect when they touch the human skin. This plant is popularly used mainly to
rheumatic pains, arthritis, ringworm and prostatic hyperplasia. The aim of this study is to evaluate the action of U.
baccifera leaves in DNA double-strand. Methods: The leaves were taken to maceration (70% ethanol) and the
ethanol was eliminated producing the crude extract (CE), that was fractionated with chloroform (CHCl3), ethyl
acetate (EtOAc) and butanol (BuOH).The genomodified capacity test used pure dsDNA (calf thymus DNA) and a
dsDNA dye (PicoGreen®). The dsDNA, diluted in a TE buffer (10mM Tris-HCl, 1mM EDTA, pH 7.5) was exposed
to CE and fractions (1000 - 1 µg/ml) and oxalic acid standard (400, 1000 and 1800 µg.ml-1), substance already
quantified in this nettle by our research group. The fluorescence was read at room temperature in a SpectraMax
M2/M2e Multi-mode Plate reader (excitation: 480nm, emission: 520 nm. Results and discussion: The most part of
U. baccifera extracts promoted damage in dsDNA. BuOH (1000 - 1 µg/ml - 59.853±0.99% - 75.180±1.55%) and
EtOAc (100 and 1 µg/ml - 92.660±4.65% - 97.042±1.19%) fractions was be classified as moderately genotoxic
(dsDNA fluorescence higher than 50%). Highly genotoxic (dsDNA fluorescence lower than 50%) were the CE
(1000 - 1 µg/ml - 26.798±0.61% - 39.233±0.19%), CHCl3 (1000 and 100 µg/ml - 26.886±5.91% and
31.777±2.92%) and oxalic acid (3.481±0.13% - 3.945±0.21%). The CHCl3 at 10 (91.325±1.50%) and 1 µg/ml
(90.289±2.15%) and EtOAc 10 µg/ml (100.89±0.55%) are statistically similar to negative control (non-treated
dsDNA sample), according to Dunnett test, presenting no genomodification capacity. Conclusion: An high
genotoxicity was observed to the extracts and fractions of the leaves of this plant, what motivated to more
studies concerning the antitumor activities of this nettle be executed.
Financial Support
Federal University of Santa Maria
Acknowledgments
-
Ethical approval
-
173
OBTAINING AND SPECTROPHOTOMETRIC EVALUATION OF CONSTITUENTS
PRESENT IN Allium sativum L. WAX
Ana Cláudia Funguetto Ribeiro
Ana Cláudia Funguetto Ribeiro
Rodrigo José Freddo
Elton Luis Denadin
Jefferson Soares
From the time of the Pharaohs, garlic (Allium sativum L.) Is used to treat various diseases. Various
volatile sulfur compounds are found in its bulb, such as aliin, ajoene and allicin. The obtaining of the garlic wax
was developed through grinding, centrifugation and heating until a greenish color wax with a characteristic scent
was achieved. The compounds were analyzed by "Headspace" and identified by gas chromatography-mass
spectrometry and NIST library and subsequently compared with commercial garlic oil. The chromatogram of the
sample containing garlic wax identified a number of sulfur compounds, such as: Diallyl disulphide, 3-Vinyl-1,2dithiacyclohex-5-ene, tetrasulfide, di-2-propenyl, 1-Propene, 3,3 ' -thiobis-, 3-Vinyl-1,2-dithiacyclohex-4-ene,
Dimethyl trisulfide, 1,3-Dithiane, sulfide, allyl and methyl hydroperoxide, 1,4-dioxan-2-yl. In the commercial
sample no volatile compound was identified. The aliin maceration process gives rise to allicin which converts
itself to various sulfur compounds, when subjected to heating, which explains the result obtained. The method of
obtaining therapeutically active compounds in the garlic bulb, proved to be effective for this protocol and may be
an alternative form of administration. Further studies will be conducted to confirm the same compounds in
different samples and microbiological testing, as well as commercial oil analysis developed by other
manufacturers aiming for a new comparison.
174
PHYTOCHEMICAL PROFILE OF STEM BARK EXTRACTS FROM Drimys brasiliensis
Miers
Andréa Michel Sobottka
Universidade de Passo Fundo (UPF)
Lenise de Bairro Antunes
Caroline Boff da Luz
Charise Dallazem Bertol
Introduction: Called as casca d'anta or cataia, the Drimys brasiliensis plant is known due to its various
medicinal uses. Phytochemical screening and thin layer chromatography (TLC) were performed with fractions
from the stem bark extracts of this species. Methods: Two extracts were obtained: extract 1 by maceration of the
bark with methanol:water (1:1) for ten days and concentration in rotary evaporator; extract 2 by turboextraction
with water for ten minutes and drying in Spray Dryer. The resulting extracts were subjected to liquid-liquid partition
with solvents: hexane (providing fractions FH1 and FH2), dichloromethane (FD1 and FD2), ethyl acetate (FAC1
and FAC2) and butanol (FB1 and FB2). The fractions were submitted to general tests to alkaloids, tannins and
flavonoids, and TLC. Results and discussion: Precipitate formation occurred with Dragendorff, Mayer, Bertrand
and Bouchardat reagents in FH1, and with the Dragendorff in FH2. It was observed the formation of precipitated
with iron salts and lead acetate in FAC1, FAC2, FB1 and FB2. Fluorescence under UV light was obtained by
oxalo-boric reaction in all fractions, being more intense in FAC1 and FAC2. Through the TLC a distinct
chromatographic profile was observed in each fraction. The developer NP/PEG for flavonoids detected fluorescent
spots in FAC1 (blue, green and yellow), FB1 (green), FD2 (weak green), FAC2 (green) and FB2 (blue). FAC1
presented similar spot to standard rutin. After the use of the developer ferric chloride for phenolic acids, dark
spots were detected on FAC1, FB1, FAC2 and FB2, characterizing these compounds. The developer KOH did not
detect coumarin. Conclusions: Different phytochemical profiles were observed between the fractions, suggesting
the presence of alkaloids in hexane fractions, tannins in ethyl acetate and butanol fractions, and flavonoids in
hexane, dichloromethane, ethyl acetate and butanol fractions, with the highest concentration in ethyl acetate and
butanol fractions.
175
CHEMICAL COMPOSITION AND IN VITRO ANTIMICROBIAL ACTIVITY OF THE
ESSENTIAL OIL OF Aloysia gratissima (GILLIES ET HOOK) TRONCOSO
(VERBENACEAE)
Anelise Levay Murari
Pharmacology Graduate Program, Federal University of Santa Maria, Santa Maria, RS, Brazil
Thiago Acosta Oliveira
Simone Cristina Benovit
Letícia Trevisan Gressler
Veterinary Medicine Graduate Program, Federal University of Santa Maria, Santa Maria, RS, Brazil
Agueda Palmira Castagna de Vargas
Veterinary Medicine Graduate Program, Federal University of Santa Maria, Santa Maria, RS, Brazil
Solon Jonas Longh
Forest Sciences Graduate Program, Federal University of Santa Maria, Santa Maria, RS, Brazil
Berta Maria Heinzmann
Pharmacology Graduate Program, Federal University of Santa Maria
Introduction: Aeromonas hydrophila, an aquatic Gram-negative bacillus, distributed globally and
ubiquitously in the natural environment, which may be implicated in a variety of human diseases. Aeromonas
species can produce various βlactamases which confer resistance to a broad spectrum of βlactams, and
therefore it is important to search for new drugs. Aloysia gratissima (Gillies & Hook) Troncoso, an aromatic plant
rich in essential oil (EO), is known in Brazil as erva-santa, and is widespread in South American folk medicine.
Methods: Plant material of A. gratissima was collected in Santa Maria, RS, Brazil (voucher specimen nº SMDB
13.077). The EO was obtained from the aerial parts by hydrodistillation in Clevenger type apparatus for 3 h
(BRITISH PHARMACOPOEIA, 2007) and subsequently analyzed by GC-MS. The evaluation of the antibacterial
activity against Aeromonas hydrophila ATCC7966 was based on the document M31-A3/CLSI/ 2008. The
concentrations tested ranged from 3,200 to 3.125 μg/mL. Results and Discussion: A total of 70 compounds were
identified in the EO. The major components were 1,8-cineole (18.54%), sabinene (9.5%), guaiol (6.79%) and
bicyclogermacrene (5.12%). The EO did not inhibit bacterial growth at the concentrations tested. Literature reports
a low susceptibility of Gram-negative bacteria to plant extracts and their components, including EO constituents.
The greatest strength of these bacteria lies in the outcome of the interaction between several factors, including
differences in their interface and a greater physicochemical complexity of the glycoprotein in their cell wall.
Conclusions: The EO is a complex blend, and some of its constituents may have antibacterial activity, while
others might antagonize this effect.
Financial Support
CAPES, HUSM/UFSN
176
Neonothopanus gardneri: ANTILEISHMANIAL ACTIVITY ASSOCIATED WITH in vitro
GROWTH INHIBITION OF Leishmania amazonensis PROMASTIGOTES FORMS
BÁRBARA CRISTINA SILVA HOLANDA QUEIROZ
FEDERAL UNIVERSITY OF PIAUI
MICHEL MUÁLEM DE MORAES ALVES
LUCAS MOREIRA BRITO
FEDERAL UNIVERSITY OF CEARA
THAYNARA PARENTE DE CARVALHO
ADRIANA CUNHA SOUZA
FERNANDO AECIO DE AMORIM CARVALHO
MARIA DAS DORES ALVES DE OLIVEIRA
JOAQUIM SOARES DA COSTA JÚNIOR
FEDERAL INSTITUTE OF PIAUI
Introduction: Leishmaniasis are zoonoses that affect approximately 12 million people worldwide and
about 1 to 2 million people are infected annually. Conventional treatments are limited and have a range of side
effects. Natural products have been investigated in the search for new alternatives associated with low toxicity.
Neonothopanus gardneri is the largest bioluminescent fungi in Brazil and is found mainly in the North and the
Northeast of the country. There are no reports of its possible biological activities. The aim of this study was to
evaluate the in vitro activity of N. gardneri extracts on promastigotes forms of Leishmania amazonensis.
Methods: Specimens were collected in the city of San Francisco, Maranhao, Brazil, then were cleaned and
frozen. Lyophilized most of the material from which was prepared the Ethyl Acetate (EtOAc ext.) and Methanolic
(MetOH ext.) extracts. With non-lyophilized part was prepared Ethanolic extract (EtOH ext.). To assess the
antileishmanial activity, 1x106 promastigotes were seeded into 96 wells plate with the extracts in concentrations
ranging from 3.200 to 12.5 μg/mL and incubated in a B.O.D. incubator at 26 °C for 48 hours. After, cell viability
was assesed by the colorimetric method of Rezasurina®. Results and discussion: The MetOH, EtOH and EtOAc
extracts were able to inhibit parasite growth. There was 100% inhibition at a concentration of 3,200 μg/mL for
AcOEt ext., and above 50% at concentrations of 3,200 μg/mL for MetOH and EtOH ext. The IC50 of MetOH,
EtOH and EtOAc ext. was 3530.57; 3112.53 and 1147.07 μg/mL, respectively. Natural products derived from
mushrooms have shown excellent results on antileishmanial activity, as described for Agaricus blazei.
Conclusions: N. gardneri showed potential to inhibit L. amazonensis promastigotes forms in vitro. Future research
should be done to explore its cytotoxic potential as well as its mechanism of action.
Financial Support
CAPES, FAPEPI
Acknowledgments
Medicinal plants research center, Laboratory of antileishmanial activity, Federal University of Piaui, Federal Institute of Piaui
177
INULINE AND WHEY AS BIOACTIVE COMPONENTS AND TECHNOLOGICAL AMORA
ICE CREAM OF CREAM (morus nigra)
Bruna Brasil Rodrigues Furtado
Fernanda Zanchet Saraiva
Faculdade Assis Gurgacz (FAG)
Luciana Oliveira de Fariña
Gabrielle Racoski Custodio
INTRODUCTION: Ice creams are high-calorie products with low nutritional value. Alternatives such as
the addition of bioactive fibers and protein sources to its formulation can improve their nutritional and functional
value, increasing acceptance and consumption. Thinking about it, the aim of this study was to develop an ice
cream with reduced fat content using inulin as fat substitute associated with whey powder aimed to improve
protein and nutritional profile while maintaining a good acceptance of the final product. METHODS: The ice cream
was prepared with traditional adjustment formula of creamy ice cream with replacement of fat per 8% inulin, 4% of
non-fat solids by whey milk powder and blackberry flavored syrup. The chemical characterization, microbiological
control was done using descriptive statistics. Sensory analysis was performed with a sample of 83 tasters of the
academic community of the institution and evaluated the acceptability index (% IA) of the product in relation to the
attributes color, texture, taste, aroma and appearance. RESULTS AND DISCUSSION: The chemical analysis
showed that the ice cream had made 33% more than the minimum required by law (2.5%) and 6% fiber, which
would lead to classification of the product as "high in fiber". The product met the microbiological requirements of
the current legislation. Sensory analysis demonstrated that the use of inulin associated to the whey powder was
satisfactory, keeping the acceptability of the product with respect to color, flavor, aroma and appearance, but did
not improve the texture of the final product. Only the texture attribute had poor acceptance rate (53.28%).
CONCLUSION: despite the good nutritional and functional quality achieved, adjustments in the components will
be needed to improve the final product texture. Financial Support: Faculty Assis Gurgacz and UNIOESTE.
Thanks: to the project team.
Financial Support
Assis Gurgacz Universitary Center and UNIOESTE
Acknowledgments
To project team.
Ethical approval
Comitê de Ética em Pesquisa do Centro Universitário Assis Gurgacz n° 141-09
178
ANTI-TRYPANOSOMAL ACTIVITY OF FLAVANONES ISOLATED FROM A PROPOLIS
SAMPLE NATIVE TO RIO GRANDE DO SUL (RS), BRAZIL
Bruna Roberta Grunwald
Aline Irala Vieira
Fabiana Gomes Nascimento Soares
Vera Lúcia Eifler-Lima
Saulo de Andrade Fernandes
Introduction: Propolis is a resinous material collected by bees (Apis mellifera) and its chemical
composition depends on the phytogeographic characteristics. Its composition is complex and diverse, being a
valuable source of different natural organic compounds with several biological activities. Some compounds such
as flavanones demonstrated activity against Trypanossoma cruzi, the causative agent of Chagas disease, one of
the major endemic diseases in Latin America. The currently available treatments are often ineffective. Therefore,
efforts in the search for new drugs are essential. Methods: A native propolis sample was purchased in the Apiary
Adams Company Comercial Exportation Ltda, Taquara, RS, Brazil. For the compounds isolation, the material was
subjected to maceration with hexane until exhaustion and then with dichloromethane. The extracts were
partitioned by column chromatography with hexane/ dichloromethane in increasing polarity. The compounds were
purified by crystallization and identified by spectroscopic methods. For the biological analysis, the epimastigote
form of T. cruzi was used, in BHI medium and fetal bovine serum. The compounds were tested at 25 μM, the
same concentration of standard drug nifurtimox (Nfx). Results and Discussion: Two main compounds were
obtained: pinostrobin (18%), and pinocembrine (5,15%). The compounds inhibited T. cruzi epimastigotes between
72.9 + 1.34% and 46.8 + 4.95, respectively. For pinostrobin, the most potent compound, an IC50 of 16.8 + 0.7 µM
was obtained. Besides, its IC50 was in the same order that Nfx (IC50=7.7 µM). Conclusions: Flavanones are not
commonly found in propolis samples from Brazil. The presence in the samples analyzed in RS can be justified by
the distinctive temperate climate, which favors the development of plants synthesizing these compounds. Further
studies are still necessary, but the isolated molecules can be considered as promising anti-T. cruzi hits.
Financial Support
CAPES, CNPq, FAPERGS
Acknowledgments
FACFAR, LRNANO
179
Box-Behnken design to optimize the phenolic content in methanolic extracts from
peels of Punica granatum L.
Bruno Sleifer Alonso
Melissa Schwanz
Introduction: The Punica granatum L. species popularly known in Brazil as pomegranate, is
characterized as a large shrub or small tree. Ethnopharmacological accounts report the use of shells as antidiarrheal, and studies show the antioxidant activity, antiviral and antibacterial, anti-diarrheal and anthelmintic.
Among the major secondary metabolites responsible for these activities, it highlights the isomers punicalagin
punicalagin A and B in the fruit peel extracts. Thus, optimization studies of extraction and analysis of phenolic
compounds shown to be important, aiming at the adequacy of the species as a drug source. Methods:
Experiments in order to investigate the most appropriate extraction conditions to optimize phenolic content of
pomegranate dry fruit peels were conducted. The method of extraction by reflux was used and three variables
were evaluated: temperature (60 - 80 ºC), mass ratio of the plant and solvent (1:75 - 1:150, m/v) and methanol
concentration (75 - 95%). The experimental design was obtained by response surface methodology Box-Behnken,
which generated 16 independent experiments. The extracts were analyzed by HPLC, and the response observed
was the sum of the areas of phenolic compounds. Results and Discussion: From the analysis of results, it was
noted that temperature and concentration parameters of the solvent influenced statistically significantly (p <0.5)
on the response assessed, positively. The variable ratio of the vegetable mass and solvent do not demonstrate a
statistically significant influence (p <0.5). Conclusions: Optimization of extraction of phenolic compounds from the
fruit peel P. granatum, using experimental design Box-Behnken indicated that the optimal parameters are found at
a temperature of 80 ° C; the mass/solvent 1: 125 w/v; and concentration of 95% methanol.
Financial Support
CAPES
180
INVESTIGATION IN VITRO TOXILOGICAL OF THE ETHANOLIC EXTRACTS FROM
PHYSALIS PERUVIANA
Carolina Ziegler Ribeiro
Universidade Regional Integrada do Alto Uruguai e das Missões - URI/Santiago
Kelly Bastos Feksa
Saionara Benvenhú Machado
Auryene Nunes do Rozário
Thaylise Vey Parodi
The plant Physalis peruviana has medicinal properties, with wide folk use as anticancer,
immunomodulatory and antibacterial and it belongs the Solanaceae family which includes about 100 species,
some related toxicity. In this sense, the in vitro test using nauplii of Artemia salina, a kind of microcrustacean
used as toxicity biomarker, is an important tool to preliminary assessment of the fraction and chemical
characterization of the extracts and optimization in vivo tests. The study aimed to verify the toxicity in a pilot test
in relation to concentration and exposure time of extracts of different plant parts. Leaves, calyx and fruit
separately were macerated with ethanol under agitation daily during a week. The final extracts were concentrated
in rotaevaporator and tested at different concentrations (0, 10, 50, 100, 150, 350, 500, 750 and 1000 µL L-1)
previously diluted in DMSO (dimethyl sulfoxide) at proportion 1:10 in order to check a survival percentage of
nauplii during 24, 48 and 72 hours (h) and the controls: saline, solution of potassium dichromate at 2% and DMSO
(in higher concentration used to dilution). Potassium dichromate showed 100% mortality after 5h and no mortality
to DMSO over time 72h. Tests revealed survival over 65% in the exposure to the leaf extract (150 µL L-1) and
rate upper to 70% for calyx extract (50 µL L-1) during 48h to both while the survival rate to the fruit extract upper
to 40% after 72h (50 µL L-1). Alkaloids and lactones are characteristic constituents of the ethanol extract of the
leaf and calyx from P. peruviana in many studies and this is the reason for the toxicity of this species. In
conclusion, preliminarily, plant chemotype of our study showed low toxicity which may be related to the
concentration and length of exposure time and not just the presence of the characteristic constituents. This work
had financial support of the Universidade Regional Integrada do Alto Uruguai e Missões - URI / Santiago.
Financial Support
181
Plinia cauliflora: DRY EXTRACT PREPARATION AND DEVELOPMENT OF MEDICATED
GUM
Carolynne Pazini Schaydegger
Pharmacy Graduate, Departament of Pharmacy and Nutrition, University of Espírito Santo, Alegre, ES, Brazil
Stela Givisiez Melo Fernandes
Wesley Careri Muller
Juliana Aparecida Severi
Professor, Departament of Pharmacy and Nutrition, University of Espírito Santo, Alegre, ES, Brazil
Janaina Cecília Oliveira Villanova Konishi
Introduction: hydroalcoholic extract obtained from Plinia cauliflora leaves (jaboticaba) was shown to be
active against Candida albicans. The present work aimed to prepare a dosage form containing this extract in order
to use it as a buccal antiseptic. Medicated gums are semisolid dosage forms that can deliver the drug into the oral
cavity. Methods: fresh leaves of P. cauliflora were washed under tap water, dried (40 oC), powdered using a knife
mill, and subjected to extraction by maceration at r.t. in 70% aqueous methanol (1:5 w/v, 21 days). The organic
solvent was evaporated at 40 °C under reduced pressure and the aqueous residue lyophilized, yielding a green
crude extract. An aliquot of this crude extract was subjected to qualitative phytochemical assays aiming to detect
the presence or absence of the main types of secondary metabolites. Moreover the solubility in water was also
evaluated. Dried extract was incorporated (5% w/w) into the medicated gums (3.5 g) and the dosage forms were
obtaining by molding under different formulations. Results and discussion: The phytochemical analysis indicated
the presence of tannins, flavonoids and saponins in the extracts, that showed be insoluble in water. The
formulations named A was prepared with gelatin, collagen and sucrose as sweetener. In the formulae named B,
the excipients were gelatin, xanthan gum, sodium carboxymethyl cellulose, microcrystalline cellulose and sodium
saccharin as sweetener. Medicated gums containing 14% w/w of gelatin (A1) showed high consistency, dificulting
the molding and the chew. In the presence of 11% gelatin, these parameters were appropriate. B2 formulation
containing 5% w/w of sodium saccharin and 14% w/w gelatin presented proper texture and pleasant taste.
Conclusions: medicated gums were properly handled on a small scale in the presence of gelatin or gelatin and
xanthan gum as thickener. The dry extract and medicated gums will be subjected to biological assays against C.
albicans strains.
Acknowledgments
The authors thank FAPES.
182
FRAGMENTATION STUDIES OF CASCAROSIDES A, B, C AND D AND TWO NEW
CASCAROSIDES BY ESI-ION TRAP MASS SPECTROMETRY
Daniel Pecoraro Demarque
Núcleo de Pesquisa em Produtos Naturais e Sintéticos, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São
Paulo, Ribeirão Preto-SP, Brazil
Danielle Rocha Pinho
Laboratório de Produtos Naturais e Espectrometria de Massas, Centro de Ciências Biológicas e da Saúde, Universidade Federal de Mato
Grosso do Sul, Campo Grande-MS, Brazil
Laboratório de Produtos Naturais e Espectrometria de Massas, Centro de Ciências Biológicas e da Saúde, Universidade Federal de Mato
Grosso do Sul, Campo Grande-MS, Brazil
Norberto Peporine Lopes
Abstract: Mass spectrometry is a powerful technique to identify organic compounds and to help in
structural elucidation. The structural elucidation by mass spectrometry must be based on fragmentation proposes,
which should be establish considering organic chemistry principles. Thus the aim of this study was isolated and
characterize fragmentation patterns of isomeric cascarosides (A/B, C/D) and two new cascarosides isomers (oxoA/oxo-B) isolated from Rhamnus purshiana (Cascara Sagrada). The powder of cascara bark was extracted by
percolation process. The crude extract (CASCrExt) obtained was dried, resuspended in methanol/water (1:1 CASMeWFas) and submitted to partition with hexane and ethyl acetate. After lyophilized, 1.5137 g of
CASMeWFas was chromatographed in classical column using silica gel and ethyl acetate: methanol as mobile
phase. The enriched fraction (672.7 mg) was fractionated in HPLC-semiprep using a C18 column and flow of 9
mL.min-1. Cascarosides were isolated, identified by NMR (1H, 13C, DEPT, NOESY) and submitted to
fragmentation study by direct infusion using an ESI-Ion Trap mass spectrometer (AmazonSL, Bruker) in positive
mode. In positive mode appears the sodiated adduct for all cascarosides. The first fragmentation was the sugar
loss (-162Da) followed by dehydration of the second sugar unit. For cascarosides A-D Retro-Diels-Alder (RDA)
reaction occurred in the sugar moiety after dehydration and the methyl acyl group was then loosed by radical
fragmentation. The new cascarosides oxo-A and oxo-B have an extra hydroxyl group in the C-10 and after RDA in
the second sugar unit occurs McLafferty-type rearrangements to form the quinone group. Differences in
fragmentation reactions were not founded between Cascarosides A-D, although the additional oxygen in C-10
allow a different fragmentation. The difference in the fragmentation pattern of the two groups of cascarosides is
the McLafferty-type rearrangement observed for the C-10 oxygenated compounds.
Financial Support
FAPESP 2014/18052-0
Acknowledgments
FAPESP, CNPq, FCFRP
183
Optimization extraction process of phenolic compounds from cranberry using
response surface methodology
Dayane Cristina de Moraes
Universidade de Sao Paulo
Sonia Aparecida Figueiredo
Michelle de Andrade
Luis Alexandre Pedro de Freitas
Maria José Vieira Fonseca
In this study, were determined the optimal conditions for phenolic compounds extraction from cranberry
powder by dynamic maceration using the surface response methodology with four independent variables: (i)
extraction time (hours); (ii) ratio plant : solvent; (iii) temperature and (iv) speed agitation (RPM) , according to BoxBehnken experimental design (4 factors and 3 levels) with analysis of three answers: total polyphenol content
(TPC); antioxidant activity (IC50; DPPH) and total solids content (TSC). The ideal conditions obtained for
extraction of cranberry powder were: extraction time 5 hours; ratio plant: solvent 1: 2; agitation speed 80 RPM
and temperature 45 ° C, corresponding to the values predicted of 433.1 mg/g; 0,100 mg/ml and 19.9% for TPC;
IC50 DPPH and TSC respectively. To check the validity of the method, the cranberry powder was extracted
under the optimized conditions. The values obtained for the answers were TPC: 418.3 mg/g ± 1.78; IC50 DPPH:
0,109 mg/mL ± 0.006 and TSC 21.88% ± 1.09. The values were similar to those predicted, indicating adequacy of
the employee model. Then, the optimized cranberry extract was evaluated with respect to its
photoprotective/photochemoprtotective activity. Preliminary results presented in this paper demonstrate the ability
of optimized cranberry extract to scavenge free radicals, with a high antioxidant potential to inhibit satisfactorily
chemiluminescence generated in the system xanthine/XOD/luminol; presenting IC50 0.202 ± 0.32 ug/ml; lipid
peroxidation using yolk egg as lipid substrate, with an IC50 of 32.6 ± 5,8ug/ml. In DPPH test, the optimized
extract showed an IC50 of 136.7 ± 6.8 mg/mL. The extract was also able to reducing UVB radiation-induced
damage at the cellular level in fibroblasts mice of L929 line, with a 41% reduction in lipid peroxidation in cells
compared to irradiated control.
184
ANTIOXIDANT ACTIVITY BY PHOSPHOMOLYBDENUM METHOD OF Erythroxylum
daphnites MART. LEAVES COLLECTED AT DIFERENTE SEASONS OF THE YEAR
Diegue Henrique Nascimento Martins
Health Science Graduate Program, Faculty of Health Sciences, University of Brasília, Brasília, DF, Brazil
Yuri Yabu Barros
Amanda de Assis Carneiro
Christopher William Fagg
Faculty of Ceilândia, University of Brasília, Ceilândia, DF, Brazil
Damaris Silveira
Pérola de Oliveira Magalhães
Yris Maria Fonseca-Bazzo
Introduction: Erythroxylum daphnites Mart., "chapadinho" or "fruto de tucano" commonly found in
Cerrado, belongs to Erythroxylaceae Family. Erythroxylum species are used as antibacterial, anti-inflammatory
and diuretic. This study aimed to investigate the antioxidant activity of aqueous extracts of E. daphnites leaves
collected at different periods of year. This specie is not well known and scientific reports are scarce, making its
research fundamental, not only as a factor of preserving biome biodiversity, but also as an identification tool and
record of this species due to its scientific potential. Methods: E. daphnites leaves were collected in Cerrado
biome - Brazil in May and August of 2013, and November, February and May of 2014. The specie was identified
by comparison with a voucher specimen deposited at the Herbarium of Universidade de Brasília under registry
Fagg CW2305. Leaves were dried at temperature of 37°C. Aqueous extracts preparation were based on the
Brazilian Pharmacopoeia 5th ed. in proportion 1:10, the material was filtered and lyophilized. Antioxidant activity
was investigated by phosphomolybdenum method. The antioxidant activity was established by comparison with
the activity of vitamin C and expressed as vitamin C equivalents. The statistical analysis were performed using
GraphPad Prism software. Results and discussion: It was observed antioxidant activity for all aqueous extract of
E. daphnites. The antioxidant capacities were expressed as vitamin C equivalents: 5.23 μg/mL (May/2013), 6.508
μg/mL (Aug/2013), 6.508 μg/mL (Nov/2013), 6.508 μg/mL (Feb/2014), 6.522 μg/mL (Feb/2014). Kruskal-Wallis
test showed no significant difference between the activity of aqueous extracts. Conclusion: E. daphnites aqueous
extract presented antioxidant activity by phosphomolybdenum method and the collection period showed no
influence on the antioxidant activity of the leaves.
Financial Support
Coordination for the Improvement of Higher Education Personnel (CAPES); National Council for Scientific and Technological Development (CNPq), Decanato de
Pesquisa e Pós-Graduação-UnB (DPP/UnB) and Fundação de Apoio à Pesquisa do Distrito Federal (FAPDF) for financial support.
Acknowledgments
Laboratório de Agroclimatologia da Fazendo Água Limpa ¿ Universidade de Brasília
185
EVALUATION OF IN VITRO RELEASE AND PERMEATION PROFILE TO TERPINEN-4-OL
OF TOPICAL CANCER SKIN THERAPY
FLAVIA LIMA RIBEIRO MACCARI
Faculty of Pharmacy - UNESP, Araraquara, SP, Brazil
FABIOLA GARCIA PRAÇA
Faculty of Pharmacy - University of São Paulo, Ribeirão Preto, SP, Brazil.
MARCO ANTONIO CORREA
MARIA VIRGINIA SCARPA
ANSELMO GOMES DE OLIVEIRA
Introduction: The terpinen-4-ol is a terpene compound produced by the secondary metabolism of several
plants, mainly of Melaleuca alternifolia, that has anti-inflammatory, antifungal and antimicrobial properties.
Recently, in vitro studies have demonstrated the effectiveness of Melaleuca alternifolia and isolated terpinen-4-ol
in the inhibition of melanoma (M14 line) and lung tumor cells (A549), breast tumor cells (MCF-7) and prostate
tumor cells (PC-3). Topical chemotherapy for the treatment of cutaneous malignancies, as basal cell carcinoma
(BCC) and squamous cell carcinoma (SCC), could be an alternative to reduce drug systemic toxicity. Topical
chemotherapy using terpinen-4-ol (TPL) could be an alternative to treat skin cancer. Methods: The aim of this
work was the development and evaluation of in vitro release and permeation profile to TPL from two different
cheap and simple medicines to topical application. TPL (relative to 1525 µg/cm2) was incorporated in either of two
formulations containing a mixture of surfactant with thickener and stabilizer in the right quantity, the difference
between them was the Cosmowax J®. TPL release rate and topical delivery were evaluated in vitro using acetate
cellulose membrane and porcine ears skin, respectively, mounted in a Franz vertical diffusion cell. Results and
discussion: Ours research demonstrated that formulation containing more hydrophilic character (Formulation A,
without Cosmowax J®.), were able to promote high amount of TPL permeated and retained in the deeper layers of
skin (where are usually located the skin cancers) at 8 hours post topical application. Conclusion: This effect the
formulation A was a simple approach to obtain higher amounts of delivery TPL into skin, which might be useful to
increase the effectiveness of topical cancer skin therapy.
Financial Support
FAPESP
186
Characterization of volatile compounds of flowers to Bouganvillea glabra choyse
Gilieli Sthefani Rodrigues Florindo
Elton Luis Gasparotto Denardin
Jefferson De Jesus Soares
Rafael Rohers
The Bougainvillea glabra choisy belongs to nyctaginacea family, are native to Latin America. This plant,
is often used to beautify streets and gardens, but according to the literature keeps a huge therapeutic potential.
Studies using leaves, have presented promising results with respect to the treatment of diabetes, diarrhea, ulcer
and bactericidal action, as well as the species has, demonstrated effect front to free radicals, one of the causes
of oxidative stress, but only few studies using flowers was observed. The object of this study was, to
characterize the volatile compounds of flowers of Bougainvillea glabra choyse, through of GCMS-HS. The flowers
were collected in March - 2016, washed with distilled water and subjected to analysis in a GCMS-HS Shimadzu
equipment. Through this analysis, were identified terpenes as phytol, 2-pentadecanone,6,10,14-trimethyl, Betulin,
3,7,11,15-tetrametyl-2-hexadecen-1-ol,
lup-20(29)-en-3-ol,acetate,(3.beta),
9,19-cyclo-9.beta-lanostane-3beta,25diol, Phytosterol as Beta-sitoesterol, Phenolic compound like vitamin E. Fatty acids like hexadecanoic
acid 2-hydroxy-1-(hydroxymethyl) ester, octadecanoic acid,2,3-dihidroxypropyl ester, n-hexadecanoic acid; methyl
17-methyl-octadecanoate, 9,12,15-octadecatrienoic acid, ethyl ester, (z,z,z). Isoprenoid like tetrahydroedulan
Glicosideos like Benzilbeta-d- glucoside, carboxylic acids like 1,2-benzenedicarboxylic acid mono(2ethylhexyl)ester. Some of these compounds present in the flowers, are already described in the literature as
potential pharmacological application. But, more tests are necessary to evaluate the efficacy pharmacology of the
Bougainvillea glabra choyse flowers.
Financial Support
UNIPAMPA,CAPES
Acknowledgments
UNIPAMPA
187
INVESTIGATION ON THE EFFECT OF ALKALOIDS FROM Lycopodiella cernua L.
(LYCOPODIACEAE) ON THE NITRIC OXIDE PRODUCTION IN THE C6 ASTROGLIAL
LINEAGE CELLS
Hellen Knecht
Universidade Federal do Rio Grande do Sul
Janaína Solomón
Larissa Bobermin
João Paulo A. dos Santos
Carlos Alberto S. Gonçalves
Eduardo Konrath
Introduction: Neuroinflammation is a part of neurodegenerative disorders such as Alzheimer's disease
(AD), and the process is triggered by inflammatory mediators (e.g. nitric oxide) released by reactive astrocytes. In
this sense, decoctions of Lycopodiella cernua are used in folk medicine to treat inflammatory conditions. The aim
of this study is to evaluate the in vitro effect of an enriched fraction of alkaloids from Lycopodiella cernua and the
isolated alkaloids cernuine and lycocernuine on the C6 astroglial lineage towards the cell viability parameters
(MTT) and the inhibition of nitric oxide production (NO) induced by LPS (lipopolysaccharide). Methods: The aerial
parts of L. cernua were subjected to an acid-base extraction to obtain an enriched fraction of alkaloids, which was
analyzed by GC-MS to allow the identification of the chemical profile. Later, the extract was subjected to
chromatographic procedures in order to isolate the compounds cernuine and lycocernuine. The enriched fraction of
alkaloids and the isolated products were tested for biochemical parameters in a C6 astroglial lineage. Results and
Discussion: After treatment on the C6 cell lineage at concentrations of 25, 100 and 150 μg/mL of the alkaloid
fraction, no significant effects on the cell viability at the MTT assay was verified, and a significant inhibition of NO
production after 24 h incubation was observed. The isolated alkaloids assayed in the concentration of 150 μM did
not induce significant changes in cell viability, but lycocernuine caused inhibition of NO production. Conclusions:
The inhibition of NO production induced by LPS associated with anticholinesterase effect previously described for
the alkaloid fraction of L. cernua makes this species promising for the treatment of AD.
Financial Support
FAPERGS
188
INHIBITION OF NOCICEPTIVE AND INFLAMMATORY RESPONSE IN MICE BY ORAL
ADMINISTRATION OF Sida tuberculata EXTRACT
Hemerson da Rosa
Andréia Caroline Fernandes Salgueiro
Morgana Duarte da Silva
Igor do Santos Coelho
Marcio Tavares Costa
Adair Roberto Soares dos Santos
Andreas S. Loureiro Mendez
Vanderlei Folmer
Introduction - Sida tuberculata (Malvaceae) is an herbaceous plant popularly used in south Brazil for the
treatment of inflammatory disorders. In order to confirm the traditional use, we aimed to assess the phytochemical
composition, analgesic and anti-inflammatory effects of the S. tuberculata extract. Methods - Dried leaves were
extracted using methanol (1:10, w,v) for 4 hours under magnetic stirrer at 40 °C. After, chemical composition was
analyzed by LC/ESI-MS system, operated in positive ion mode. For in vivo assays, dried extracts were evaluated
using experimental chemical nociception induced by intraperitoneal acetic acid (AA) or subplantar formalin
injection and by carrageenan-induced (i.p) visceral inflammatory process. The animals (Swiss mice) were orally
pretreated with different doses of extract 1hour before the induction. After injection of AA or formalin, was
recorded the number of writhing (20 min) and time spent licking or biting the injected hind paw (0-5 min neurogenic
phase and 15-30 min inflammatory phase) respectively. For carrageenan assay, was investigated cell migration
and myeloperoxidase activity in peritoneal exudate. Results and Discussion - Phytochemical characterization
detected phytoecdysteroids derivatives as major compounds in extract. In addition, extract (from 10 mg/kg)
significantly reduced the amount of writhing assessed by AA and decreased the licking time of the injected paw in
two phases of formalin test. In peritonitis, extract at 10 mg/kg concentration reduced the polymorphonuclear
leukocytes influx and decreased myeloperoxidase activity. Therefore, these results suggest a possible
contribution of phytoecdysteroids to the analgesic and anti-inflammatory effect observed. Conclusions - Our
findings suggest the leaves extract of S. tuberculata are able to inhibit acute inflammation and nociception on
chemical behavioral models, confirming the traditional use of this plant.
Financial Support
Capes, CNPq, UNIPAMPA
Acknowledgments
UNIPAMPA, UFSC, UFRGS
Ethical approval
027/2014
189
EVALUATION OF ANTIOXIDANT ACTIVITY IN LIPOSOMES OF GUARANÁ (Paullinia
cupana var. sorbilis).
Isabel Roggia
Centro Univeristário Franciscano
Ana Júlia Figueiró Dalcin
Gerson Fernandes de Brum
Kátia Nascimento
Michele Rorato Sagrillo
Ivana Beatrice Mânica da Cruz
Patrícia Gomes
Introduction: Guarana is one of the most promising compounds of brazilian flora. It has many
therapeutic properties including antioxidant activity. When Guarana is exposed to unfavorable environmental
conditions, degradation of actives compounds, loss of stability and/or effect may occur. The nanotechnology can
be an alternative path to protect drugs against the degradation. Thus, the objective of this study was to determine
the antioxidant activity of liposomes containing 1 mg.mL-1 of guarana. Methods: The antioxidant activity was
evaluated by DPPH radical scavenging activity method. The guarana powder was supplied by Embrapa. Guarana
samples were prepared in raw material (RM), white liposomes (no guarana) produced by reverse phase
evaporation method (BRF), ethanol injection (BEI), liposomes containing 1 mg.mL-1 of guarana powder prepared
by reverse phase evaporation method (LRF) and ethanol injection (LEI). The samples were evaluated at
concentrations of 1, 10, 25 and 50 mg.mL-1. The absorbance was determined in spectrophotometer at wavelength
of 518 nm. The assay was performed in triplicate. The calculation of the antioxidant activity was determined by
percentage of inhibition of DPPH. Furthermore, it was determined of IC50. The DPPH solution was used as a
negative control. Results and discussion: White liposomes (BRF and BEI) did not show antioxidant activity in
different concentrations. For liposomes LRF, LEI and RM, scavenger activity of DPPH at the concentration of 1
mg.mL-1 was not observed. For other concentrations, increasing activity with increasing concentration of the
compound was observed. Conclusions: Liposomes of guarana were prepared by LRF and LEI showing
scavenging activity and may be an antioxidant potential, but further studies are necessary for a more precise
conclusion. The LEI liposomes when compared to LRF showed higher scavenging activity. Likewise, the
scavenging activity in the LEI was similar to RM.
Financial Support
CAPES
Acknowledgments
190
SUSCEPTIBILITY EVALUATION OF ETIL ACETATE EXTRACT OF Hyptis mutabilis
(RICH.) BRIQ. AGAINST BACTERIA CAUSING FOOD-BORNE DISEASE
Isadora Aguirre Rosa
Faculty of Pharmacy, Universidade Regional Integrada do Alto Uruguai e das Missões-URI, Santiago, RS, Brasil
Cristiele Fiuza Soares
Faculty of Pharmacy, Universidade Regional Integrada do Alto Uruguai e das Missões-URI, Santiago, RS, Brasil
Vanessa Oliveira Domingues
Faculty of Pharmacy, Universidade Federal de Santa Maria, Santa Maria, RS, Brasil
Thaís Felli Kubiça
Health Sciences Departament, Faculty of Pharmacy, Universidade Regional Integrada do Alto Uruguai e das Missões-URI, Santiago, RS,
Brasil
Sydney Hartz Alves
Microbiology and Parasitology Departament, Universidade Federal de Santa Maria, Santa Maria, RS, Brasil
Brasil
Introduction: The incidence of bacterial infections has increased over the last years and is considered
an important cause of mortality due to bacterial resistance to conventional drugs. Thus, new antimicrobial agents
have been researched in plants. The species Hyptis mutabilis (Rich.) Briq. (Lamiaceae), known as cidreira-defolha and sambacoite, is used to treat flu, gastrointestinal diseases and as antiseptic. Its essential oil showed
antiulcerogenic, antimicrobial and sedative effects. This study evaluated the antibacterial activity of ethyl acetate
extract of H. mutabilis against bacteria causing food-borne disease. Methods: Aerial parts were collect in March
2013 in Santa Maria (voucher n° SMDB 13076, UFSM) and submitted to sequential extraction (dichloromethane,
ethyl acetate, methanol 70%) using a Soxhlet apparatus during 12 hours. The antibacterial activity was carried out
using the broth microdilution method as established by M07-A9 protocol (CLSI, 2012) in triplicate. The extract was
tested in concentrations range of 2000-3,906 μg/mL. Results and Discussion: The ethyl acetate extract showed a
Minimal Inhibitory Concentration (MIC) of 2000 μg/mL against Salmonella enteritidis ATCC 13076. The strains of
Bacillus cereus ATCC 14579, Escherichia coli ATCC 25422 and Salmonella pullorum ATCC 9140 was not
inhibited by the extract (MIC > 2000 μg/mL). Previous research with essential oil of this plant demonstrated
antibacterial activity against B. cereus by diffusion technique, while E. coli showed resistance. The differences
between results could be related to distinct samples, methodologies and bacterial strains used. Conclusion: The
antibacterial activity of ethyl acetate justifies partially the popular plant use to treat gastrointestinal diseases, such
as salmonellosis. However, the involvement of constituents of other polarities should not discarded.
Financial Support
Secretaria de Desenvolvimento Econômico, Ciência e Tecnologia (Convênio SCIT 73/2013), FURI-URI and Conselho Nacional de Dese nvolvimento Científico e
Tecnológico (CNPq)
Acknowledgments
The authors are grateful to Secretaria de Desenvolvimento Econômico, Ciência e Tecnologia (Convênio SCIT 73/2013) and FURI-URI for financial support. The
author I.A. Rosa is grateful to Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) for her their undergraduate scholarship.
191
CYTOTOXIC EVALUATION OF ESSENTIAL OIL ANACAUÍTA (Schinus molle L.) IN CELL
CULTURE OF HUMAN LYMPHOCYTES.
Jonathaline Apollo Duarte
Léa Augusta de Bairros Zambrano
Elizandra Gomes Schmitt
Luciane Dias Quintana
Mariana Balhego Rocha
Luís Flavio Souza de Oliveira
Michel Mansur Machado
The Anacauíta (Schinus molle L.) has also been employed in popular medicine to treat various diseases
standing out its use in particular for the treatment of respiratory infections. Despite its wide use in therapy, there
are no studies in the literature focused on its possible cytotoxic effects, demonstrating the importance of our
study. For this it was performed the extraction of essential oil of leaves in nature by hydrodistillation in Clevenger
apparatus for eight hours. After extraction determine the LD50 of essential oil in the culture of human lymphocytes
using proliferation assay (Neubauer chamber) in agreement with previous studies. Thus, the negative control was
considered (PBS pH 7.2), positive control (Bleomycin 3mg/mL) and test (LD50, LD50/10, LD50/100, LD50/1000
and LD50/10000). All data were statistically analyzed by ANOVA followed by "post hoc" Bonferroni (p<0.05). The
LD50 was found to be 30μg/mL The positive control showed more than 80% damage compared to the negative
control. Already the concentrations of 30 and 3μg/mL damaged in 70% and 17.71% respectively. However, the
lowest concentrations expressed counteraction once the concentration of 0.3μg/ml promoted greater than 95%
proliferation, while those of 0.03 and 0.003μg/mL favored over 100% proliferation. Based on the results, observed
that the concentrations of 30 and 3μg/mL demonstrated lower cytotoxic activity than the positive control,
however, 0.3, 0.03 and 0.003μg/mL did not affect cell proliferation, characterizing a possible protective effect of
essential oil at low concentrations. Thus, it is evident the need for further research to assess toxicity and
bioavailability corroborating to the safety of the plant in question.
192
CHEMICAL AND BIOLOGICAL STUDIES OF ORGANIC EXTRACTS OF ROOTS OF
SCHINOPSIS BRASILIENSIS
Juceni Pereira David
Universidade Federal da Bahia (UFBA)
Jorge Mauricio David
Bruno Oliveira Moreira
Mariluze Cruz
Regiane Yatsuda
Introduction Schinopsis brasiliensis Engl. (Anacardiaceae) is an endangered species which is used in
Brazilian folk medicine to treat impotence, sore throat, cough and diarrhea. This work describes the
phytochemical study and antioxidant evaluation, lethality of brine shrimp and, anticholinesterase, antinociceptive
and anti-inflammatory activities of the organic extracts of the roots. Methods The CHCl3, EtOAc and BuOH
extracts were obtained from the partition of the crude MeOH extract of the roots of S. brasiliensis. The EtOAc and
BuOH extracts were submitted to different chromatographic techniques to isolate the secondary metabolites. The
isolates were identified by different spectrometric techniques. The extracts and pure compounds were submitted
to in vivo Writhing Test and Neutrophils migration, and in vitro antioxidant (DPPH and carotene/linolenic acid),
AChE s and BST tests. Results and Discussion The MeOH extract of S. brasiliensis showed good antioxidant
activity and the CHCl3 and EtOAc extracts showed inhibition of AChE (68.77 ± 1.20/86.91 ± 1.76%) and in BST
(IC50<500 ppm). From the extracts were isolated and identified sitosterol, daucosterol, α/βamyrin, gallic and
ellagic acids, quercetin3OβDxylopyranoside, the new chalcone dimer 2', 4, 4', 5-tetrahydrohydroxychalcone-(27',8-8')-4'-hydroxyetenylbenzene besides (7"*R, 8"*S)-2', 4, 4', 5-tetrahydroxychalcone-(2-7",8-8") - 2''', 4", 4'''trihydroxy- 7", 8"-dihydrochalcone. The evaluation of the antinociceptive and anti-inflammatory activities showed
that both extracts were actives at doses of 50 and 25 mg/kg. Conclusions From the roots of S. brasiliensis were
isolated a new chalcone dimer and the extracts and isolated compounds were evaluated in biological and
pharmacological assays. They permitted to indicate this plant is a potential species for further biological and
pharmacological studies.
Financial Support
CNPq and FAPESB
Ethical approval
Ethical Committee of for Animal Research of the University of Uberaba (protocol 0107/2009)
193
THE ROLE OF DIOSMIN IN GLIOBLASTOMA CELLS
Juliana Moreira Soares
ANTONIO GOMES SOARES
EMBRAPA
LUCIANA FERREIRA ROMÃO
Introduction: Glioblastoma is a high malignant tumor characterized by rapid cell proliferation, tissue
invasion and production of angiogenic factors. Despite advances, treatment possibilities are still not efficient and
many studies seek for alternatives to control tumor growth. In this context, flavonoids are identified as natural
substances capable of acting against some tumors. Flavonoids are polyphenolic compounds extracted from
plants. Some of them have an antitumor effect and could induce apoptosis on tumor cells by interacting with
proteins that control the cell cycle and change expressed genes of the tumor. Thus, they can act on proliferation,
differentiation, angiogenic, inflammation pathways and in the metastatic evolution. Methods: This study evaluates
the effect of the flavonoid diosmin on Glioblastoma cells GBM95, GBM02 (CONEP number 2340) and U87 in
order to search new anticancer compounds. The cells were cultured in medium DMEM-F12 with 10% of fetal
bovine serum, until reach confluence. The MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)
assay were performed for the analysis of the cells viability after the treatment with diosmin in different
concentrations (10, 50, 75, 100 and 150 μM) and time (24, 48 and 72 hours). The Tunel assay (terminal
deoxynucleotidyl transferase) was performed to analyze DNA fragmentation after the treatment with diosmin 100
μM for 24 and 48 hours. Results: Treatment with diosmin reduced viability of tumor cells GBM95, GBM02 and
U87 in 24 and 48 hours. Preliminary results from Tunel assay suggest that diosmin induced GBM95 cell death by
DNA fragmentation in 24 and 48 hours. Additional studies are still required to determine the effect of diosmin on
cell proliferation and death. Conclusion: In summary, the role of diosmin, was evaluated on Glioblastoma
progression and viability but the mechanism of action of this flavonoid still need to be investigated.
Financial Support
FAPERJ and CNPq
Ethical approval
CONEP number 2340
194
ANTIBACTERIAL ACTIVITY OF MENTHOL AND ESSENTIAL OILS OF Mentha sp.
GROWING IN HYDROPONIC CULTIVE.
Kelly Bastos Feksa
Aline Monteiro Moscato
Thaís Felli Kubiça
Brasil.
Eduardo Pereira Shimóia
Faculty of Agronomy, Universidade Regional Integrada do Alto Uruguai e das Missões-URI Câmpus Santiago, Santiago, RS, Brasil.
Vânius Ventorini Veiga
Faculty of Agronomy, Universidade Regional Integrada do Alto Uruguai e das Missões-URI Câmpus Santiago, Santiago, RS, Brasil.
Brasil.
Introduction: Aromatic plants Mentha piperita e a Mentha spicata (Lamiaceae) are known as hortelãpimenta (peppermint) e hortelã-peluda (spearmint), respectively. These species are used as carminative,
antispasmodic, antiseptic and antipruritic. Their pharmacological effects, such as antimicrobial activity, have been
related to the presence of essential oils and their compounds. Thus, the aim of this study was to evaluate the
antibacterial potential of the essential oils obtained from these plants grown in hydroponic culture. Furthermore,
antibacterial activity of menthol was also evaluated. Methods: Aerial parts of M. piperita and M. spicata grown on
the Campus of URI, Santiago, were collected in June and July 2014, dried in a ventilated oven at 40ºC and
submitted to hydrodistillation procedure by 3 hours in a Clevenger-type apparatus. The antibacterial activity was
carried out using the broth microdilution method as established by M07-A9 protocol (CLSI, 2012) in triplicate. The
samples were tested in concentrations range of 2000-3,906 μg/mL. Results and Discussion: The yields of the
essential oils obtained from M. piperita and M. spicata were 0.69 ± 0.03% e 0.16 ± 0.03 % (Mean ± SEM),
respectively. Only Staphylococcus aureus ATCC 29213 and Escherichia coli ATCC 25422 were inhibited by
essential oils with minimal inhibitory concentration (MIC) of 2000 µg/mL. The strains of Enterococcus faecalis
ATCC 29212 and clinical isolated of Staphylococcus saprophyticus and Salmonella pullorum were not inhibited by
the essential oils tested (MIC > 2000 µg/mL). Menthol did not show activity against the strains tested. The lack of
menthol activity, when tested alone, can be due necessity of synergistic effect of other essential oil compounds.
Conclusion: Mentha sp. cultivated in hydroponic systems do not produce essential oils with significant
antibacterial activity.
Financial Support
FURI-URI
Acknowledgments
The authors K.B. Feksa and E.P. Shimóia are grateful to PIIC-URI for her their undergraduate scholarship.
195
ANTIFUNGAL ACTIVITY OF PROANTHOCYANIDIN POLYMERIC TANNINS FROM
Stryphnodendron adstringens ON Candida non-albicans AND SYNERGISTIC EFFECT
IN COMBINATION WITH FLUCONAZOLE
Kelly Ishida
Departamento de Microbiologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo/SP, Brasil
Aline Luiza Duarte de Freitas
Departamento de Microbiologia, Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo/SP, Brasil
Departamento de Farmácia, Centro de Ciências da Saúde, Universidade Estadual de Maringá, Maringá/PR, Brasil
Celso Vataru Nakamura
Departamento de Ciências Básicas da Saúde, Centro de Ciências da Saúde, Universidade Estadual de Maringá, Maringá/PR, Brasil
Stryphnodendron adstringens is often used in the Brazilian folk medicine as astringent, cicatrizing, and
to treat vaginal infections. Candida albicans is the main etiological agent of candidiasis, however Candida nonalbicans (CNA) species had increased in the last decades, mainly those resistant to the antifungals. In previous
studies we showed that fractions (F2 and F2.4) rich in proanthocyanidin polymers extracted from S. adstringens
interfere with the growth of C. albicans isolates. Thus, the aim of this work is to determine the antifungal effect of
F2 and F2.4 obtained from S. adstringens stem bark on CNA species and to evaluate the sinergistic effect of F2
or F2.4 with standard antifungals (Amphotericin B - AMB, fluconazole - FCZ, and miconazole - MCZ). The
antifungal susceptibility of CNA strains was assessed by the broth microdilution assay; and the antifungal
combination was performed using checkerboard technique and the interaction was defined by the Fractional
Inhibitory Concentration index (FIC index). Clinical isolates of C. glabrata were more susceptible to F2 and to
F2.4 (1.9 - 3.9 µg/mL) followed by C. parapsilosis and C. krusei (7.8 - 31.2 µg/mL). In contrast, C. tropicalis
isolates presented lesser susceptibility to F2 and to F2.4 (7.8 - 125 µg/mL). Additionally, isolates resistant to AMB
and/or FCZ and/or MCZ were susceptible to F2 and to F2.4. Both F2 and F2.4 showed fungistatic activities
against CNA strains. All antifungal drug interactions tested against Candida spp. were considered indifferent
(0,5<FICI?4), except subfraction F2.4 (0.49 µg/ml) combined with FCZ (1 µg/mL) on C. glabrata ATTC 2001 (FIC
Index = 0.5) featuring as synergistic effect. Taken togheter, our results indicate that the proanthocyanidin
polymeric tannins presents into the fraction F2 and subfraction F2.4 from Stryphnodendron adstringens could be
an alternative to candidiasis treatment caused by the non-albicans species alone or in combination with FCZ.
Financial Support
CAPES, CNPq, FAPESP
196
PHYTOCHEMICAL PROFILE AND ANTIOXIDANT ACTIVITY OF ESSENTIAL OILS FROM
Schinus weinmannifolius AND Schinus polygamus
Krissie Daian Soares
UFRGS
Leticia D. Jacobi
Sérgio A. L. Bordignon
La Salle University Center, Canoas, RS, Brazil
Miriam A. Apel
The genus Schinus belongs to the family Anacardiaceae and studies have shown anti-inflammatory,
antimicrobial and antioxidant activities to species of this genus. One of the secondary metabolites in this genus
are the essential oils that have been the target in search for new substances with antioxidant action. This study
aimed the chemical characterization and antioxidant activity of the essential oil from leaves and fruits of Schinus
weinmannifolius and leaves of Schinus polygamus from Rio Grande do Sul. The essential oils were obtained by
hydrodistillation in Clevenger apparatus for 4 hours. The chemical analysis of leaves and fruits was carried out by
gas chromatography coupled to mass spectrometry (GC/MS). The antioxidant activity of the essential oils was
determined by DPPH radical scavenging assay and ability to inhibition lipid peroxidation (TBARS method), using
the BHT and Rutin as positive control, for both assays. The yield of essential oil from leaves and fruits of S.
weinmannifolius was 0.5% and 2.0%, respectively, and 0.4% for leaves of S. polygamus. The major compounds
identified in the leaves of S. weinmannifolius were αcadinol (21.2%), spathulenol (11.0%) and cubenol (9.8%). In
the fruits αcadinol (20.5%), spathulenol (9.9%) and limonene (9.8%). The main components for the leaves of S.
polygamus were n-nonane (33.5%) and αcadinol (15.4%). In the DPPH assay none of the species of leaves and
fruits presented antioxidant activity in different concentrations. For the leaves and fruits of S. weinmannifolius and
S. polygamus, the results of the inhibition of lipid peroxidation (TBARS) assay was 59% in the concentration of
250µg/mL, for both the species tested. Schinus, essential oils, antioxidant Acknowledgements: Financial support
from Capes.
Financial Support
CAPES
197
PHARMACOKINETIC ABSORPTION MODELING OF FLAVONOID CHRYSIN
Larissa Lachi Silva
Elza Kimura
Andréa Diniz
Flavonoids are secondary metabolites widely distributed in plants, commonly present in human food and
dietary supplements. Chrysin belongs to subclass flavone and presents aintioxidant and anticancer activities.
Propolis and honey have high levels of chrysin and it is correlated to their pharmacological/biological effects. In
spite of many activities described, the pharmacokinetic parameters remain poorly explored and barely
characterized. The aim of this study was to estimate the intestinal absorption pharmacokinetic profile of chrysin
by closed-loop intestinal perfusion (Dalouisio's model). Chrysin purchased from Sigma (purity 97%). For the
perfusion, male Wistar rats (n=6) were implanted with an intestinal cannula, all experiment were approved by the
Ethical Committee of University of Maringa (protocol nº010/2011). The solution (10 mL) containing chrysin (1.0
mg/kg) was introduced directly into the small intestine and samples (300 µL) of this solution in the intestinal
lumen were collected at 0; 1.5; 3; 5; 10; 15; 20; 25 and 30 minutes by a syringe attached to the cannula. The
chrysin remaining amount in intestinal perfusate was analyzed by HPLC-UV. Data were analysed by nonlinear
mixed effects model using software Monolix432s. The final percentage absorbed (%Abs) was determined by
application of five models for the absorption data set. The first order absorption was the most suitable for this data
set, different from the more complex model suitable for another c-glycoside, vicenin-2, which involved degradation
and efflux constants. The %Abs was more than 45%. The absorption constant (ka) was estimated as 0.0214 min1(±0.0005). This pharmacokinetic model was efficient to describe decreasing of intestinal concentration of chrysin
after to apply simple animal surgery and quantification techniques.
Financial Support
Fundação Araucária
Ethical approval
Ethical Committee of University of Maringa (protocol nº010/2011)
198
ANTIDERMATOPHYTIC EFFECT OF Schinus lentiscifolius ESSENTIAL OIL AND ITS
SYNERGY WITH TERBINAFINE AND CICLOPIROX
Letícia Jacobi Danielli
Ana Júlia Maciel
University of Rio Grande do Sul, Porto Alegre, RS, Brazil
Bruna Pippi
Agricultural and Environmental Microbiology Graduate Program, University of Rio Grande do Sul, Porto Alegre, RS, Brazil
Sérgio Augusto de Loreto Bordignon
La Salle University Center, Canoas, RS, Brazil
Miriam Anders Apel
Introduction: Increasing the resistance, limited therapeutic arsenal and high levels of toxicity, justify the
search for new substances as an alternative to antifungal conventional therapy, such as compounds isolated from
plants. Essential oils are one of the most promising groups of compounds for the development of new
antimicrobial agents whose activity is widely reported. Thus, the aim of this study was to evaluate the antifungal
activity of S. lentisicifolius essential oil and determine its synergistic effect, combined with classical antifungal
drugs. Methods: The oil was obtained by hydrodistillation with subsequent identification of the chemical
composition by GC-MS. Antifungal activity against filamentous fungi, was determined by minimum inhibitory
concentration (MIC), while effects of the oil combined with terbinafine and ciclopirox were evaluated by fractional
inhibitory concentration indices (FICIs) from checkerboard method and time-kill assay. Results and Discussion:
Chemical analysis of oil presented the predominance of sesquiterpenes, with γeudesmol (12.8%), elemol
(10.5%), βeudesmol (10.2%), βcaryophyllene (9.9%) and αpinene (8.2%) as major compounds. The oil exhibited
effect against dermatophytes - Trichophyton rubrum, Microsporum canis and M. gypseum, with MICs around 125
to 500 µg/mL. The antifungal activity of combinations of oil with terbinafine and ciclopirox, indicated synergistic
and additive effects, with a FIC index under or equal 0.75. Combination of subinhibitory concentrations of
essential oil and terbinafine demonstrated greater fungal inhibition growth compared to ciclopirox. Time-kill curve
method confirmed efficacy of oil and terbinafine combination, with reduction in fungal counts after 6h. Conclusion:
The observed synergistic effect between essential oil and the antifungals indicates an opportunity for the
development of promising strategies to improve the conventional therapy.
Financial Support
Capes
199
EXTRACT OF Litchi chinensis LEAVES EXERTS PHOTOPROTECTIVE EFFECTS
AGAINST UVA AND UVB IRRADIATION IN MURINE FIBROBLASTS (L929)
Liliani Carolini Thiesen
Universidade do Vale do Itajaí
Christiane Meyre Silva Bittercourt
Angelica Garcia Couto
Tania Mari Bellé Bresolin
José Roberto Santin
Introduction: The identification of novel compounds that exert a biological UV-protective effect in skin
cells represents an important strategy for preventing ultraviolet (UV)-induced skin damage. The L. chinensis (LC)
leaf soft extract (5% herbal drug: solvent/ethanol 70°GL), previously quantified by LC-UV (33.8 and 56.6 mg/g of
epicatechin and procyanidin A2, respectively) presents promissory effects, taking into account its antiradicalar
and anti-inflammatory properties. The present study aimed to assess the photoprotection effects of LC extract.
Methods: The sun protection factor (SPF). Photoprotection was evaluated by exposure L929 cells to UVA (1.1,
2.20 or 3.3 J/cm2) or UVB (0.014, 0.021 or 0.035 J/cm2) irradiation. The cells were treated with the LC extract (1,
10 or 100 μg/mL) for 24 hours, after this period, the cells were rinsed and exposed to UVA or UVB irradiation to
measure the viability and ROS production by MTT and DCFH-DA methods, respectively. Safety evaluation was
complemented by agarose overlay-test. Results: The results showed that LC extract presents UVA and UVB
absorption profile, and the SPF was 18.9 ± 0.4. The extract reduced the UVA and UVB-induced cytotoxicity in
L929 cells. Data from UVA irradiation showed that the extract (10 μg/mL) promotes 80% and 65% of cell viability
at 1.1 J/cm2 and 2.2 J/cm2 irradiation, respectively. The untreated cells already showed viability of 49% and
40%, respectively. In the UVB irradiation the extract showed photoprotective effect in all concentrations in
different irradiation doses, while the untreated cells showed a viability of around 50%. Furthermore, the extract
protected L929 cells against reactive oxygen species produced by UVA and UVB irradiation. It was not observed
irritative effect exert by LC. Conclusion: Data obtained show that LC extract presents photochemoprotective
effect against UVA and UVB irradiation with promising potential applications.
200
ANTINOCICEPTIVE ACTIVITY OF STACHYS LAVANDULIFOLIA VAHL. (LAMIACEAE)
ESSENTIAL OIL IN MICE
LUCINDO QUINTANS JUNIOR
ROSANA S.S. BARRETO
JULLYANA S.S. QUINTANS
MARCIO R.V. SANTOS
MARCELO C. DUARTE
IRWIN R.A. MENEZES
HENRIQUE D.M. COUTINHO
Universidade Regional do Cariri (URCA)
GOKHAN ZENGIN
Selcuk University, Konya, Turkey
ABDURRAHMAN AKTUMSEK
Selcuk University, Konya, Turkey.
ADRIANO A.S. ARAÚJO
Stachys lavandulifolia is a medicinal plant widely used in Turkey folk medicine due its pharmacological
property, but little is known about its essential oil. So, we assessed the antinociceptive effect of S. lavandulifolia
essential oil (EOSl) and ()αbisabolol (BIS), main compound, in orofacial nociceptive behavior induced by
algogen in mice. Experimental protocols were approved by the regulatory bodies of the Institutional Animal Care
and Use Committee at UFS/Brazil (CEPA/UFS # 72/2015). The GC-FID/GC-MS assessment of EOSl showed
presence of ()αbisabolol (56.4%), bicyclogermacrene (5.3%) and δcadinene (4.2%) as the main compounds.
Male Swiss mice were pretreated with EOSl (25 or 50 mg/kg, p.o.), BIS (25 or 50 mg/kg, p.o.), (morphine (3
mg/kg, i.p.) or vehicle (saline 0.9% with two drops of tween 80, 0.2%), before formalin- (20 µl, 2%), capsaicin- (20
µl, 2.5 µg) or glutamate- (20 μL, 25 μM, s.c.) injection into the right upper lip (perinasal area) in mice. We showed
that p.o. treatment with EOSl or BIS displayed significant inhibitory (p<0.05 or p<0.01 or p<0.001) effects in
different orofacial pain tests on mice, but BIS proved to be most effective reducing significantly the nociceptive
behavior in all tests including both phases of the formalin test. The analgesic effect is not related to any
abnormality since EOSl- or BIS-treated mice exhibited no performance alteration on the grip strength meter.
Moreover, EOSl and BIS exhibited significant reduction (p<0.05 and p<0.01) of the pro-inflammatory cytokine
TNFα. Together, our data corroborate the traditional medicine use of S. lavandulifolia to analgesic applications.
Financial Support
CNPq and FAPITEC/SE
Acknowledgments
FAPITEC/SE, CNPq, CAPES and FINEP
Ethical approval
CEPA/UFS # 72/2015
201
PRESENCE OF QUERCETIN DERIVATIVES IN HYDROETHANOLIC EXTRACTS
OBTAINED FROM Cuphea glutinosa CHAM & SCHLTDL (LYTHRACEAE)
Marí Castro Santos
Introduction: Popularly known in Brazil as "sete-sangrias", Cuphea genus is used in folk medicine in the
form of infusion, as diuretic, antipyretic, anti-inflammatory and anti-hypertensive. The chemical composition of
some Cuphea species has been reported, presenting triterpenes, steroids, fatty acids, carbohydrates, tannins and
flavonoids, mainly quercetin derivatives. Aims: The present study aimed to investigate the species Cuphea
glutinosa Cham & Schltdl, determining its chemical composition using UPLC-ESI-MS. Methodology: The leaves
were selected and processed by exhaustive maceration using ethanol 40% (v/v). The chromatographic analyses
were performed on an Acquity® UPLC system (Waters Co., MA, USA) equipped with detectors by DAD-UV and
mass spectrometer Q-TOF. The assay was conducted using a reverse-phase system, by fast C18 analytical
column Shim-pack XRODS; mobile phase of acetonitrile:methanol (4:1, v/v) and water with 0.1% formic acid (pH
3.0); elution by gradient; flow of 0.2mL/min; injection volume of 5.0 µL. Mass spectrometry analyses were
conducted in positive-ion mode (ESI+). Results and Discussion: The chromatographic assay was able to
separate the chemical constituents, allowing the identification by MS detector. Through a detailed evaluation of
the mass spectra, four different quercetin derivatives could be suggested: quercetin-3-O-glucoside (m/z 465),
quercetin-3-arabinoside (m/z 435), quercetin-3-glucuronide (m/z 479) and isorhamnetin (m/z 317). The presence of
phenolic compounds and flavonoids in Cuphea is reported in some works, with emphasis in the chemical
derivatives of kaempferol, myricetin and quercetin. Conclusions: Based on our results, it is suggested that
chemical composition of C. glutinosa leaves is predominant in flavonoids, mainly quercetin derivatives. These
findings contribute to understand the chemical profile of this species, prospecting new researches with
phytochemical focus.
Acknowledgments
FAPERGS; PBDAUNIPAMPA
202
Evaluation of Antinociceptive Activity of Methanolic Fractions of Sugarcane Juice
(Saccharum officinarum L.)
Mariana Alves Soares
Natália Linhares Coutinho Silva
Laboratório de Estudos em Farmacologia Experimental - FF/UFRJ
Anne Caroline Candido Gomes
Laboratório de Fitoquímica - IPPN/UFRJ
Anne Katherine Candido Gomes
Naomi Kato Simas
Ricardo Machado Kuster
Ana Luísa Palhares de Miranda
Laboratório de Estudos em Farmacologia Experimental - FF/UFRJ
Jorge Luiz Mendonça Tributino
Laboratório de Farmacologia Molecular - ICB/UFRJ
Different sugarcane (Saccharum officinarum L.) segments are popularly used for the treatment of
pathological conditions. Some components identified in the sugarcane stalk juice (SSJ) are flavonoids (FL)
derived from apigenin, luteolin and tricine, for which there are biological actions described, such as antiinflammatory activity. Furthermore, some FL are described as ligand of opioid receptors, key pharmacological
targets for analgesia. The methanol fraction of natural (SJ) and fermented (FSJ) sugarcane juice were obtained
from reverse chromatography of the SSJ (SP71-1406) which has high concentration of FL. Considering the
actions described for isolated FL and extracts of plants rich in FL, the ease of obtaining the sugar cane juice as a
source of FL and the need to develop new approaches for the treatment of painful disorders, we evaluated the
analgesic potential of SJ and FSJ (100mg/kg po) using animal models of nociception and inflammation: writhing
induced by acetic acid 0.1N (i.p.), nociception induced by formalin 2.5% (i.pl.), paw edema induced by
carrageenan 1% (i.pl.) and hot plate assay for evaluation of thermal hyperalgesia (CEUA FARMACIA01/02/03).
SJ shows a 41% reduction in the number of writhes compared to the control group, while FSJ shows 31%
reduction, which points to an analgesic effect of these substances. In the formalin test, SJ and FSJ reduce the
licking time only in neurogenic phase, with inhibition of 44.2% and 56.7%, respectively. This effect was reversed
by naloxone, which shows that their analgesic effect is related to modulation of the opioid system. SJ and FSJ
not significantly reduced either paw edema or the carrageenan-induced thermal hypersensitivity. Conclusion:
These results indicate analgesic effect for SJ and FSJ, which is related to modulation of the opioid system
without significant activity on inflammatory components. This work contributes to the ethnopharmacological study
of Saccharum officinarum, potential source of FL.
Financial Support
FAPERJ, CAPES, PIBIC/UFRJ
Ethical approval
CEUA FARMACIA 01/02/03
203
DETERMINATION OF ISOFLAVONE AGLYCONES AND FURANIC COMPOUNDS IN
SOYBEAN EXTRACTS AND GENOTOXICITY EVALUATION OF THE MAIN IMPURITY
FORMED AFTER ACID HYDROLYSIS PROCESS
Pharmacy Graduate Program, University of Rio Grande do Sul, Porto Alegre, RS, Brazil
Juliana da Silva
PostGraduate Program in Cell and Molecular Biology Applied to Health (PPGBioSaude), Lutheran University of Brazil (ULBRA), Canoas,
RS, Brazil
Jaqueline Nascimento Picada
RS, Brazil
Ivana Grivicich
RS, Brazil
Ana Letícia Hilario Garcia
RS, Brazil
Débora Kuck Mausolff Papke
RS, Brazil
Martin Steppe
Introduction: Soybean extraction followed by acid hydrolysis is an usual process to obtain bioactive
isoflavones. However, the furanic compounds hydroxymethylfurfural (HMF) and ethoxymethylfurfural (EMF) are
also formed during the process. These compounds can be considered impurities and must be assessed when the
intention is the pharmaceutical use of the soybean acid hydrolyzed extract (SAE). In this context, the goals of this
study were the development and validation of an ultra-fast liquid chromatography (UFLC) method for determination
of isoflavones (daidzein, glycitein, genistein) and impurities (HMF, EMF) in SAE, as well as the evaluation of the
in vitro genotoxicity of the major impurity (EMF) present in the extract. Methods: Soybeans were defatted,
extracted with ethanol 80% and acid hydrolyzed. The samples were neutralized, diluted and analyzed by the
validated UFLC method, using a C18 column (100 x 2.0 mm i.d.; 2.2 µm) and gradient system of acetonitrile and
water with 0.1% trifluoroacetic acid. The EMF genotoxicity in HepG2 cells was performed by the alkaline comet
assay at non-cytotoxic concentrations. Results and Discussion: The UFLC method showed to be linear, specific,
precise, accurate and robust in the range of 0.1 - 10.0 µg/mL for isoflavones and 10.0 - 100.0 µg/mL for furanic
compounds. After the hydrolysis process, each 100.0 g of defatted soybeans can produce about of 0.20 g of
isoflavone aglycones and 2.0 g furanic compounds. During the EMF toxicity studies, concentrations between 0.1
to 1.3 mg/mL were not cytotoxic for HepG2 cells after 3 h of exposure, but induced a significant DNA damage,
mainly by oxidative stress mechanisms. Conclusions: An UFLC method was validated for the determination of
bioactive isoflavones and impurities present in SAE. The major impurity (EMF) can induce DNA damage in
HepG2 cells. Therefore, purification procedures to remove this compound from the SAE are recommended during
healthcare products development.
Financial Support
This work was supported by the Brazilian funding agencies CNPq and FAPERGS. MCN wishes to thank CAPES for her scholarship.
204
Antineoplastic Evaluation of Limonium brasiliense (Boiss) Kuntze (Plumbaginaceae)
Fernanda Gomes Beserra
Gabriela Souto Vieira de Mello
Andressa Blainski
Maringa State University
Maringa State University
Ivan da Rocha Pitta
BACKGROUND: Limonium brasiliense is an herb that inhabits salt marshes on Brazilian coast and
southern states. Rhizomes of L. brasiliense often contain tannins that are responsible for some biological
responses including antineoplastic activity. Thus, this works aims to evaluate antineoplastic activity of L.
brasiliense rhizomes crude extract (CE) and ethyl-acetate fraction (EAF). METHODS: L. brasiliense rhizomes
were acquired in Rio Grande do Sul, Brazil in 2013. CE was prepared using a mixture of acetone: water (70:30,
v/v) 10% extractive solution and the remaining material were lyophilized. Cytotoxic assay with healthy donors
Peripheral Blood Mononuclear Cells (PBMCs) were performed by MTT to verify if these plant materials are
cytotoxic to normal cells. CE and EAF were added in a 96-well plate containing 5.5 x 10E05 cell/well at 10, 50,
100 and 200 µg/mL. After 72h MTT was added and the absorbance was measured at 570nm after 24h. HEP-G2
(hepatocarcinoma), T47-D (breast cancer) and HL-60 (acute promyelocytic leukemia) viability was also evaluated
by MTT assay. IC50 (µg/mL) was calculated by OringPro8 and the Selectivity Index (SI) by the ratio between
PBMC IC50 and Tumor Cell IC50. RESULTS AND DISCUSSION: CE and EAF were less toxic at PBMCs
(IC50=96.78±3.75; 75.82±10.81, respectively). CE was toxic to T47-D (IC50 = 90.68±10.64; SI = 1.08) and HL-60
(IC50 = 61.69±8.5; SI = 1.56). EAF was toxic to HEP-G2 (IC50 = 59.47±5.55; SI = 1.27) and HL-60 (IC50 =
17.26±0.28; SI = 4.39). CE and EAF were considered as promisors when SI values were higher than 3. CE and
EAF were previously analyzed and showed phenolic compounds in CE and condensed tannins (gallocathechin
and epigallocathechin) in EAF composition. Antineoplastic activity of tannins has been reported and these results
can be associated with these molecules at CE and EAF composition. CONCLUSION: CE and EAF of L.
brasiliense rhizomes showed an antineoplastic activity at HEP-G2, T47-D and HL-60 cells.
Financial Support
CNPq, CAPES, INCT_if.
Acknowledgments
Therapeutic Innovation Postgraduate Program, Federal University of Pernambuco.
Ethical approval
CEP CCS-UFPE: 1.285.288
205
HEALING OF Mimosa scabrella Benth EFFECT
Matheus Martins Falcão
Franciscan University Center, Santa Maria, RS, Brazil.
Ilaine Lourdes Hanauer
Gerusa Paz Porto
Camila Franco
Gisele Scotti do Canto
Federal University of Santa Maria, Santa Maria, RS, Brazil
INTRODUCTION: Pressure ulcers (PU) reach 2.7 to 29.5% of patients admitted to hospitals, with a
prevalence of 66% in elderly patients with femoral neck fractures, 33% in critically ill patients and 40% in
bedridden patients with spinal cord injuries. Among the consequences of them, are increased hospitalization
period, increased risk of deadh at 4.5 times and elevated use of economic resources. Considering these
consequences, the difficulty of access and the high cost of treatments, this study aims to identify the healing
potential of the plant Mimosa scabrella Benth which is commonly used as healing. METHODS: This study was
conducted with 24 adult male Wistar rats, which were subjected to a skin lesion of 1.5 cm². These animals were
treated for 8 days with 3% Hydroxypropyl Methyl Cellulose gel containing or not Mimosa scabrela Benth extract
at concentrations of 12% and 36%. In this period, the lesion was measured and evaluated daily for crusts and
hypertrophic scar development. On the seventh day was carried biometric evaluation of pH of lesion. The
experimental protocol was approved by the Institutional Animal Ethical Committee (number 004/2014). RESULTS
AND DISCUSSION: It was found that there was a reduction in the size of the lesion in addition to an acceleration
in the development of crusts and formation of hypertrophic scarring in animals that were treated with 3%
Hydroxypropyl Methyl Cellulose gel containing Mimosa scabrella Benth extracts although the extracts did not
reduce the pH of the lesion these animals, which indicate a preventive effect of bacterial colonization. The
hydroxypropyl Methyl Cellulose gel had not effect per se on size of lesion and did not influence the effect of the
extract, but increased the pH from the injured skin. CONCLUSION: This study showed that the extract of
Mimosa scabrella Benth has a potential healing effects. Additional studies involving evaluation of histological
parameters are needed to confirm this effect
Financial Support
Ethical approval
number 004/2014
206
PARAQUAT CAUSE LOCOMOTOR DEFICIT AND DECREASE DOPAMINE LEVELS IN
Drosophila melanogaster: PROTECTIVE ROLE OF Bougainvillea glabra choisy
LEAVES EXTRACT
Mayara Batú Gonçalves
Jefferson de Jesus Soares
Matheus Bianchini
Mateus Cristofari Gayer
Rafael Roehrs
Elton Luis Gasparotto Denardin
The herbicide paraquat (PQ) has been increasingly reported in epidemiological studies because of their
neurotoxic effects. This compound causes destruction of neuronal cells, causing a deficiency in the production of
dopamine which increases the risk of developing neurodegenerative diseases, including Parkinson's disease
(PD). Thus, PQ is widely used to induce symptoms of PD in different experimental models including Drosophila
melanogaster (DM). The fly fruit DM has been used as an alternative animal model for screen of natural
therapeutic agents for the treatment of PD. Plants possess many bioactive compounds that may be useful for
slowing down or preventing the onset of PD symptoms. Bougainvillea glabra choisy (BG) is a plant species of the
Nyctaginaceae family, originally from southern Brazil. Literature reports indicate that BG leaves have antiinflammatory, anti-ulcer and anti-hyperglycemic activities. The objective of this study was to evaluate the
neuroprotective effect of ethanolic extract of BG leaves against the neurotoxicity induced by paraquat in
Drosophila melanogaster. The ethanolic extract (EE) was prepared by maceration for 15 days. Thereafter, the
extract was filtered concentrated on rotaevaporator and store at -18 ºC until use. Flies (male, 1-3 days old) treated
with PQ and EE were divided into four groups: (1) control; (2) PQ; (3) EE (4) PQ + EE. PQ and EE were added
into the food's flies at final concentration of 3,5 mM and 120μL/mL, respectively. After 96 h, the negative
geotaxis, open field and analysis of dopamine content in the head of flies were performed. Flies treated only with
PQ showed locomotor deficits and had lower dopamine content compared to the control group. The co-treatment
with EE reduced the locomotor deficits and avoided the reduction of dopamine levels caused by paraquat. The
flies had a locomotor activity similar to flies in the control group clearly showing a neuroprotective effect of the
EE.
Financial Support
UNIPAMPA, CAPES
Acknowledgments
UNIPAMPA
207
ADMINISTRATION OF THE HYDROALCOHOLIC YACON ROOT AND LEAF EXTRACT
DO NOT ALTER THE HEPATIC AND RENAL FUNCTIONS OF DYSLIPIDEMIC RATS.
Micaela Federizzi de Oliveira
Patrícia Martinez Oliveira
Ritiéle Pinto Coelho
Alessandra Melise Golke
Bruna Cocco Pilar
Angélica Aparecida Güllich
Lyana Feijoó Berro
Melissa Schwanz
Vanusa Manfredini
Introduction: A plant that has been highlighted due to its medicinal properties is the Yacon (Smallanthus
sonchifolius), that have been linked to beneficial effects on human health. Studies have shown that the Yacon
have hypoglycemic activity, however little is known about other properties. This study evaluated the liver and
kidney function in hypercholesterolemic rats treated with hydroalcoholic Yacon root and leaf extract. Material and
Methods: For this study it were used male wistar rats, divided into 6 groups of 6 rats each: G1: normal diet
(healthy) (NaCl 0,9%); G2: hypercaloric diet (control) (NaCl 0,9%); G3: Yacon leaf extract 20mg/Kg (YLE20); G4:
Yacon leaf extract 40mg/Kg (YLE40); G5: Yacon root extract 20mg/Kg (YRE20); G6:Yacon root extract 40mg/Kg
(YRE40). All experiments were approved by the Ethics Committee on Animal Use Experimentation of the Federal
University of Pampa, CEUA, Uruguaiana, Rio Grande do Sul, Brazil (Protocol 034/2013). The lyophilized extracts
were administered once daily by gavage for 14 consecutive days. Enzymatic markers for liver function (aspartate
transaminase - AST, alanine transaminase - ALT) and renal (creatinine, uric acid) were determined by automated
equipment (A25 Biosystem) for in vitro diagnostics. Results and Discussion: Serum biochemical markers
evaluating the hepatic) and kidney function of the animals were within the normal limits. There were the significant
differences (p <0.05) In These parameters. Conclusion: Analysis of serum biochemical markers Evaluating the
renal and hepatic function of the animals demonstrated, during the study period, que this extract was not
hepatotoxic or nephrotoxic.
Financial Support
FAPERGS, CNPq and CAPES.
Ethical approval
Approved by the Ethics Committee on Animal Use Experimentation of the Federal University of Pampa, CEUA, Uruguaiana, Rio Grande do Sul, Brazil (Protocol
034/2013).
208
QUANTITATIVE AND QUALITATIVE VARIATION FOR VOLATILE OILS OF Piper
gaudichaudianum, Piper regnellii AND Piper umbellatum POPULATIONS FROM RIO
GRANDE DO SUL
Michele Andréia Rambo
Krissie Daian Soares
Sérgio Augusto de Loreto Bordignon
Centro Universitário La Salle
Miriam Anders Apel
Introduction: The Piperaceae family belongs to Piperales order and is one of the earliest families of
Angiosperms (1). The Piper genus comprises about 2000 species and only 12% of their species were studied
from the chemical point of view (2). Objective: The objective of this work is the analysis of the chemical
composition of the volatile oil from leaves of Piper gaudichaudianum, Piper regnellii and Piper umbellatum native
populations of Rio Grande do Sul. Methods: The aerial parts of both species were collected in Três Cachoeiras. A
voucher specimen was deposited in the Herbarium of the Universidade Federal do Rio Grande do Sul (ICN). The
oil from the leaves of the species was obtained by hydrodistillation, using the Clevenger-type apparatus for 4
hours. The chemical analysis was carried out by gas chromatography coupled to mass spectrometry (GC/MS).
The identification of compounds was based by comparison of both retention indices and mass spectra with
authentic samples and data from literature (3). Results and Discussion: The yield of volatile oil from leaves of P.
gaudichaudianum was 0.4%, P. regnellii was 0.5%, and for P. umbellatum was 0.1%. The major compounds
identified in the leaves of P. gaudichaudianum were characterized by the monoterpenes hydrocarbons αpinene
(32.2%) and βpinene (67.9%). P. regnellii were the sesquiterpenes dill apiole (16.7%) and apiole (42.0%) and for
P. umbellatum were the sesquiterpenes βcaryophyllene (25.2%), germacrene D (36.4%) and bicyclogermacrene
(12,2%). Conclusion: These results show that populations of P. regnellii and P. umbellatum derived from the
biosynthetic pathway of the group caryophyllane while P. gaudichaudianum were characterized by the presence of
monoterpenes, which are derived from geranyl pyrophosphate initial precursor of monoterpenes, that after some
cyclization reactions giving rise to compounds of structural type pinanos (αpinene and βpinene).
Financial Support
Capes
Acknowledgments
Capes, CNPQ
209
CYTOSTATIC AND CYTOTOXIC EFFECTS OF CONVALLATOXIN IN A549 CELLS
Naira Fernanda Zanchett Schneider
NAIRA FERNANDA ZANCHETT SCHNEIDER
LARA CRUZ PERSICH
LUCAS LOURENÇO MAROSTICA
RODRIGO MAIA PÁDUA
WOLFGANG KREIS
Friedrich-Alexander Universität
FERNAO CASTRO BRAGA
CLAUDIA MARIA OLIVEIRA SIMOES
Introduction: Despite significant progress in oncology therapeutics in the last decades, the urge to
discover new cytotoxic agents still remains. Recently, several groups have shown the antitumor properties of
cardenolides against cancer cells and in several in vitro and in vivo assays. In this sense, the aim of this study
was to evaluate the cytostatic and cytotoxic effects of the cardenolide convallatoxin (CON) on human non-smallcell lung cancer cells (A549). Methods: To evaluate CON effects in A549 cells several assays were carried out.
Trypan blue exclusion assay, cell cycle analyses and clonogenic capacity were used to evaluate proliferation and
metabolism effects. Phosphatidylserine exposition by annexin-V/PI assay and DAPI nucleus staining were used
to evaluate induction of cell death. Results and Discussion: Concerning to the Trypan blue assay, A549 cells
proliferation and viability reduction was observed after CON treatment at 10, 50 and 100 nM. The reproductive cell
integrity assay to establish colonies showed cells that almost complete loss their clonogenic capacity. Cell cycle
analyses after 24, 48 and 72 h showed G2/M arrest and accumulation of cells in subG0, mainly at 72 h. These
effects on cell cycle were accompanied by modulation of cyclins B1, D1 and p53 expression, also observed at
100 nM. Additionally, annexin-V positive cells and DAPI staining evaluation suggested 30% of cell death by
apoptosis. The nucleus shrinkage and fragmentation induced by CON was attenuated by a pan-caspase inhibitor
indicating that at least part of the ffects caused in A549 cells was dependent of caspase. Taken these results
together, the strongest effects on cell death was observed at 100 nM in 24h, while at 10 nM a significant cell
death was observed at 72 h. Conclusion: In conclusion, these results suggested that CON was able to stop cell
proliferation by inducing cell death in A549 cells, thus overcoming multiple anti-cell death mechanisms present in
cancer cells.
Financial Support
CAPES and CNPQ
210
PHYTOCHEMICAL CHARACTERIZATION OF Cunila microcephala benth INFUSION:
PILOT STUDY
Natacha Cossettin Mori
UNICRUZ
Roberta Cattaneo Horn
Gabriela Tassotti Gelatti
Tamiris Felippin
Ana Caroline Tissiani
Mariana Spanamberg Mayer
Diego Pascoal Golle
Jana Koefender
Candida Elisa Manfio
Nídia Ledur Müller Castro
Introduction: The Cunila microcephala Benth is a creeping herb belonging to the Lamiaceae family and
popularly known as "poejo". The infusion of the leaves and flowers are empirically used as a stimulant, antispasmodic and in the treatment of respiratory infections and chronic cough. In view of the current situation, we're
increasing research to the application of this plant for the treatment, prevention and cure of diseases. Besides the
widespread popular use of "poejo", it is important to investigate the antioxidant potential of this plant through the
phytochemical investigation. Methods: Fresh leaves of Cunila microcephala Benth from the garden of the Cruz
Alta University (UNICRUZ), Rio Grande do Sul, were used for the preparation of infusion 50g/L. The
measurement of total polyphenols, flavonoids and condensed tannins were performed by the method described by
Chandra & Mejia (2004), Woisky & Salatino (1998) and Morrison et al. (1995), respectively. The results were
expressed by media ± standard deviation. Results and discussion: It was found a concentration of 1.42±0.025g of
gallic acid equivalents/g of fresh plant, 0.69±0.015 g of quercitin equivalents/g of fresh plant and 0.124 ±0.0015g
of catechin equivalents/g of fresh plant. This results corroborated with Felisbino (2014) studies, that revealed the
presence of phenolic compounds in Cunila microcephala Benth extract. Conclusion: The presence of antioxidant
phytochemicals in "poejo" infusion indicates that this plant has an important antioxidant potential and may
eventually present as a therapeutic alternative for the conditions of oxidative stress.
211
TOXICITY OF Urera baccifera GAUDICH (URTICACEAE) ROOTS IN LEUKOCITES
Pâmila Pinheiro da Fontoura
Pharmacy Undergraduate program, Integrated Regional University of High Uruguay and Missions, Santiago Campus, Santiago, Rio Grande
do Sul, Brazil.
Michel Mansur Machado
Postgraduate program of Pharmaceutical Sciences, Federal University of Pampas, Uruguaiana, Rio Grande do Sul, Brazil
Margareth Linde Athayde
Postgraduate program of Pharmaceutical Sciences, Federal University of Santa Maria, Santa Maria, Rio Grande do Sul, Brazil
Sydney Hartz Alves
Postgraduate program of Pharmaceutical Sciences, Federal University of Santa Maria, Santa Maria, Rio Grande do Sul, Brazil
Pharmacy Undergraduate program, Integrated Regional University of High Uruguay and Missions, Santiago Campus, Santiago, Rio Grande
do Sul, Brazil; Postgraduate program of Pharmaceutical Sciences, Federal University of Santa Maria, Santa Maria, Rio Grande do Sul,
Brazil
Introduction: Popularly known as "Urtigão", Urera baccifera (Urticaceae) leaves and roots infusions are
used to treat skin and urinary infections. Oxalic acid was already quantified in this species by our research group.
Methods: The roots were collected in São Francisco de Assis, Brazil (May, 2010). The material was taken to
maceration (70% ethanol). The ethanol was eliminated producing the aqueous extracts which part of was
completely dried (crude extract - CE), and part were fractionated with chloroform (CHCl3), ethyl acetate (EtOAc)
and butanol (BuOH). Leukocyte cultures were prepared using samples collected from human donors by vein
puncture. The blood collected was immediately transferred to a culture medium containing 1 ml of RPMI 1640,
supplemented with 10% fetal bovine serum and 1% streptomycin/penicillin. The cells were placed in an incubator
at 37°C in a microaerophilic environment for 72 hours. Each sample (extract and fractions) and 0.8 mg/ml oxalic
acid (Sigma standard), diluted in PBS buffer, was added to the blood to get a final concentration of 100 μg/ml.
The amount of total leukocytes (proliferation) was determined by counting in a Neubauer chamber. Results and
Discussion: Oxalic acid (OA) caused a 30.7% reduction in the leucocyte proliferation and 15% leucocyte deaths
and the BuOH displayed the highest values in these assays (24.15% and 5.0%, respectively), followed by the CE
(2.33% leukocyte deaths).The mitotic index in the leukocyte culture was lower in the BuOH fraction of roots
(8.3%), similar to the OA index, which was 6.0%, considering the negative control. However, there were no
statistically significant differences between these treatments. Conclusion: BuOH presented the highest quantities
of OA and displayed results comparable with the standard, indicating that this compound can be responsible for
the toxic effects of the plant and could explain its stinging effect.
Financial Support
Federal University of Santa Maria and Federal University of Pampas
Ethical approval
23081.005770/2009-38
212
EVALUATION BODY WEIGHT GAIN IN RATS WITH HYPERCHOLESTEROLEMIA
SUPPLEMENTED WITH YACON ROOT (SMALLANTHUS SONCHIFOLIUS)
Patricia Martinez Oliveira
Ritiéle Pinto Coelho
Angélica Aparecida Güllich
Bruna Cocco Pilar
Patrícia Maurer
Micaela Federizzi de Oliveira
Jacqueline da Costa Escobar Piccoli
Vanusa Manfredini
Introduction: Yacon (Smallanthus sonchifolius) has been considered a food with multiple functions
because it has bioactive compounds que may have beneficial effects on human health. Most of the benefits of
consuming Yacon have been attributed to its content of fructooligosaccharides (FOS), resisting the hydrolysis of
digestive enzymes and fermented in the colon. This study investigated the effects of Yacon root extract on body
weight gain in rats with hypercholesterolemia. Material and Methods: The animals were divided into: G1: Normal
diet (healthy) (NaCl 0.9%); G2: hypercaloric diet (control positive); G3: Oral suspension of simvastatin 10 mg / kg
(SIM); G4: Yacon root extract 20 mg / kg (YRE20); G5: Yacon root extract 40 mg / kg (YRE40); G6: YRE20 +
SIM; and G7: YRE40 + SIM. The lyophilized extracts were administered once daily by gavage for 14 consecutive
days. Body weight registration of the animals was performed during all experimental periods. All experiments were
approved by the Ethics Committee on Animal Use Experimentation of the Federal University of Pampa, CEUA,
Uruguaiana, Rio Grande do Sul, Brazil (Protocol 034/2013). Results and Discussion: At the end of the
experiment, all groups showed body weight gain, but there was a small weight gain in animals supplemented with
the Yacon root extract compared to G2. Conclusion: Yacon root controlled a weight gain of animals suggesting
that this effect may be due to the high content of FOS.
Financial Support
Ethical approval
Approved by the Ethics Committee on Animal Use Experimentation of the Federal University of Pampa, CEUA, Uruguaiana, Rio Gran de do Sul, Brazil (Protocol
034/2013)
213
BACTERICIDAL AND MORPHOLOGICAL EFFECTS OF ISOCOUMARIN PAEPALANTINE
ON Helicobacter pylori
Rodrigo Rezende Kitagawa
Federal University of Espirito Santo
Joao Paulo Loureiro Damasceno
Otalibio Castiglioni Nunes
Rita de Cassia Ribeiro Gonçalves
Introduction: Helicobacter pylori, a bacterium that infects half the world's population, is recognized as
the main cause of chronic gastritis and peptic ulcers. Helicobacter pylori infection induces an inflammatory
response and changes in the gastric micro-environment. Host immune cells, especially neutrophils and
macrophages, release inflammatory mediators and large quantities of reactive oxygen and nitrogen species (ROS
and RNS, respectively), which are related to an increased risk of gastric cancer. In the present study, we
evaluated the anti-H. pylori activity and effects on bacterial morphology of the isocoumarin paepalantine isolated
from the capitula of Paepalanthus bromelioides. Methods: The antibacterial activity was assessed by the
spectrophotometric broth microdilution method, and the studies of bacterial morphology were assessed by
scanning electron microscopy. In this study we used minimal inhibitory concentration (MIC) and sub-MIC
paepalantine on H. pylori. Results and Discussion: Paepalantine exhibited significant anti-H. pylori activity at a
minimum inhibitory concentration (MIC) of 128 μg/mL and at a minimum bactericidal concentration (MBC) of 256
μg/mL, and scanning electron microscopy revealed significant morphological changes of the bacterial cell. The
MIC and sub-MIC of the paepalantine induced blebbing and sphere formation, and pronounced filamentation in H.
pylori. These modifications suggest the interaction of paepalantine with Penicillin-Binding Proteins (PBPs), which
are associated with the helical form of the bacterium. Conclusions: The results of the present study indicate that
paepalantine is a promising compound for additional studies involving the pathogenesis of the gastric injury
caused for H. pylori, as studies in vivo.
Financial Support
CAPES and FAPES
Acknowledgments
Laboratory of Cell Ultra Structure/CCS/UFES
214
NEW SEMI-SYNTHETIC BETULIN DERIVATIVES AND THEIR ANTICANCER AND ANTIINFLAMMATORY ACTIVITY
Sayonara Maria Calado Gonçalves
Gloria Narjara Santos da Silva
Simone Cristina Baggio Gnoatto
Ivan da Rocha Pitta
INTRODUCTION: The betulin is a lupane from pentacyclic terpenes, obtained from plants of the
species Betula sp. It is a class of compounds studied extensively because they have a range of biological
activities including anti-inflammatory and anticacer. Our interest in this study with new betulin derivatives was to
determine the cytotoxic activity against neoplasic cell lines and their effects on the modulation levels of
inflammatory cytokines. METHODOLOGY: Cytotoxic MTT test was used for tumor cell lines prostate cancer
(DU145), and breast cancer (MCF-7) and peripheral blood mononuclear cells (PBMC) in different doses (1, 10 and
100µM). The cytokines IFN and IL-17 were measured by ELISA from the culture supernatant of healthy
volunteers. RESULTS AND DISCUSSION: The novel derivatives obtained from betulin had IC50> 100 in PBMC
and tumor cell lines DU145 and MCF-7. However betulin showed IC50 = 49.36 (DU145) and 45.21 (MCF-7). The
positive control, doxorubicin was not toxic to normal cells and possessed a high selectivity for neoplastic cells
contrary to betulin and their derivatives, where it was not possible to calculate the SI, since it was not possible to
find the IC50 of these molecules. The derivative 2a was able to significantly reduce cytokine INFg at 100μM, as
well as new derivative 2c had an inhibitory effect on IFNg all tested doses. The cytokine IL-17 production was
reduced by the new derivative 2e at a dose of 10μM. CONCLUSION: In this study we conclude that the new
derivatives of betulin did not have cytotoxic activity against tumor cell lines. However, some new derivatives
were able to reduce the production of inflammatory cytokines INF and IL17 in PBMC.
Financial Support
INCT_if and FACEPE
Acknowledgments
Postgraduate Program in Therapeutic Innovation, Federal University of Pernambuco, 2Organic Synthesis Laboratory of Phytochemistry, Federal University of
Rio Grande do Sul
Ethical approval
CEP-CCS-UFPE: 1.351.357.
215
Evaluation of hypoglycemic activity of aqueous extract of bark of C. speciosa in Wistar
rats with hypercholesterolaemia induced by Tyloxapol
Thiane Martins Messina
Mylena da Silva Vargas
Natiéle Figueiredo da Rosa
Francieli Moro Stefanello
Universidade Federal de Pelotas (UFPel)
Fabiane Moreira Farias
Weliton Baldin
Introduction: Ceiba speciosa (Malvaceae) is a tropical tree species with broad geographic distribution.
The tea of its bark is used in the northwestern region of Rio Grande do Sul for reducing and maintaining blood
levels of cholesterol both by healthy people and those with high cholesterol levels. Despite the regional popularity
of this species, scientific literature shows no pharmacological data on it, but indicates hypoglycemic activity for a
taxonomically related species (Ceiba pentandra). Thus the purpose of this study was to evaluate the glucose
levels in wistar rats with hypercholesterolaemia induced by Tyloxapol treated with C. speciosa. Methods: This
study was approved by the Ethics Committee on Animal Use of UFPel under number 5747/2015. The aqueous
extract of C. speciosa was obtained by decoction and lyophilisation. For the assessment of glucose levels, the
animals were treated with the aqueous extract of C. speciosa (200mg/kg and 400mg/kg) for 14 days.
Subsequently, hypercholesterolaemia was induced by intraperitoneal administration of Tyloxapol (200mg/kg), and
48 hours after the animals were sacrificed, their blood was collected and glucose levels were evaluated. Results:
The aqueous extract of C. speciosa (200 mg/kg) was able to promote a statistically significant reduction (p ≤
0.05) in blood glucose levels in treated animals compared to positive control group that received only induction of
Tyxolapol. Although population uses this species for reducing cholesterol levels, the results obtained in this
experiment are important to given that many individuals with hypercholesterolemia also exhibit hyperglycemia.
Conclusion: Considering the intense regional use of this species as a therapeutic resource further investigation of
possible hypoglycemic activity of C. speciosa is necessary having in mind that the literature indicates
hypoglycemic activity for C. pentandra.
216
VITAMIN C CONTENT IN TUBERS CREM
Vanessa Bernardi Braga
Program of Graduate Studies in Plant Science from the Federal University of Rio Grande do Sul, Porto Alegre, RS, Brazil.
Ingrid Bergman Inchausti de Barros
Program of Graduate Studies in Plant Science from the Federal University of Rio Grande do Sul, Porto Alegre, RS, Brazil.
Jean Carlo Machado
Faculty of Agronomy of the Federal University of Rio Grande do Sul , Porto Alegre, RS, Brazil.
The crem (Tropaeolum pentaphyllum) it is an unconventional food , native to southern Brazil and show
versatility of uses: its leaves and flowers are eaten in salads and its reserve organ, known as " crem potato ",
grated and added to red wine vinegar is used as a traditional condiment for meats and soups. On the other hand,
crem is used in popular therapies to prevent vitamin C deficiency, body detoxification and hypercholesterolemia.
As the condiment has stood out for its taste, aroma and bioactive compounds. The present study aimed to
evaluate the vitamin C content of the crem tubers and compares it with data from the scientific literature for other
condiments derived from storage organ. The determination was carried out at the Laboratory of Horticulture and
Forestry of the UFRGS. The material used was a sample of tubers (varying in sizes and weights) acquired from
the Public Market of Porto Alegre, RS, Brazil. Tubers were washed, sliced and grated for obtain a small material
fragments. The analyzes of vitamin C were performed in triplicate by the method of 2,4-dinitrophenylhydrazine (
DNPH ). The average content showed 18.61 mg of vitamin C per 100 g crem tubers. Comparing with literature
data for other seasonings prepared similarly, it is apparent that this content can contribute significantly to the
nutritional intake from crem tubers. But the vitamin C content of crem is far below of vitamin C horseradish
content (178mg / 100g). Therefore, from the results it was possible to elucidate the vitamin C content in crem
tubers with unpublished data.
Acknowledgments
CAPES, FAURGS
217
HPLC ANALYSIS AND FREE RADICAL ACTIVITY OF Persea americana LEAF EXTRACT
AND FRACTIONS
Viviane Cecilia Kessler Nunes Deuschle
Mariana Tatsch
Ritiel Corrêa da Cruz
Regis Augusto Norbert Deuschle
Carine Viana Silva
Persea americana, popularly known as the avocado, belongs to the Lauraceae family. The objective of
this research is to identify and quantify the phenolic compounds in Persea americana leaves extract and relate
them to their anti-radical action. The extracts obtained were: hydroethanolic, ethyl acetate, and n-butanol. High
performance liquid chromatography (HPLC-DAD) was performed in reverse phase. All chromatographic operations
were carried out at ambient temperature, and in triplicate. Assessment of the ability to scavenge free radicals in
relation to the superoxide radical is based on the generation of radical O2 - throughout the system using an
enzymatic reaction catalyzed by the enzyme hypoxanthine xanthine oxidase. For hydroxyl radicals were
performed using the 2-deoxy-D-ribose method. The results indicated the presence of the following compounds:
hydroethanolic extract: chlorogenic acid (Tr = 20.208): 9.86 mg/mL ± 0.11 and rutin (Tr = 32.548): 11.21 ± 0.18
mg/mL; Ethyl acetate extract: chlorogenic acid (Tr = 20.194): 0.37 ± 0.009 mg/mL, quercetin (Tr = 41.020): 3.77 ±
0.029 mg/mL and rutin (Tr = 32.548): 2.20 ± 0.02 mg/mL; For the Butanolic extract: chlorogenic acid (Tr = 20.185):
8.72 ± 0.04 mg/mL, quercetin (Tr = 40.975): 6.13± 0.066 mg/mL and rutin (Tr =32.487): 20.68 ± 0.05 mg/mL. The
inhibition of superoxide and hydroxyl radicals was determined only for the hydroethanolic extract. The results were
superior to the superoxide anion (81% ± 0.013) as compared to the hydroxyl radicals (75 ± 0.06%). This ability to
inhibit free radicals, shown by the extract demonstrates direct relationship to the identified phenolic compounds,
since they have known antioxidant activity, from the literature.
218
ANTIOXIDANT ACTIVITY FROM Erythroxylum daphnites MART. AQUEOUS EXTRACT
AND FRACTIONS
YRIS MARIA FONSECA
Eduardo de Jesus Lima Nepomuceno
DIEGUE HENRIQUE NASCIMENTO MARTINS
Yuri Yabu de Barros
Christopher William Fagg
Faculty of Ceilândia, University of Brasília, Ceilândia, DF, Brazil
Pérola Oliveira Magalhães
Dâmaris Silveira
Erythroxylum daphnites belongs to Erythroxylaceae family, and it is popularly known as "chapadinho" or
"fruta-de-tucano". It can be found in Cerrado. Erythroxylum species has been used as antibacterial, antiinflammatory and diuretic. This study aims to evaluate antioxidant activity and obtain fingerprint by HPLC of E.
daphnites leaves aqueous extract and fractions. Plants were collected in August and November (2013) and
February (2014) in Cerrado. A sample was deposited at Herbarium of Universidade de Brasilia (UB) under the
voucher number Fagg CW2305. Aqueous extracts were prepared based on the Brazilian Pharmacopeia 5th ed.
The fractionation of crude aqueous extract (CAE) was performed using hexane and ethyl acetate as solvent to
obtain two fractions (F1-ethyl acetate fraction and F2-aqueous fraction). The extracts fingerprint was obtained by
HPLC-DAD. Antioxidant activity was investigated by DPPH assay (ascorbic acid and rutin were used as positive
control). The statistical analysis was performed using GraphPad Prism software. The statistical analysis (ANOVA
one-way, followed Tukey) showed significant difference (p<0.0001) between IC50 values of CAE (7.96 μg/mL), F2
(6.79 μg/mL) and positive controls (2.69 μg/mL ascorbic acid and 3.42 μg/mL rutin). However, F1 (3.29 μg/mL)
showed no significant difference (p<0.0001) compared to positive controls. F1 showed the highest activity when
compared to CAE and F2. The CAE fingerprints obtained by HPLC-DAD showed phenolic compounds (flavonoid
type). The F1 fraction was the most active and contain a high concentration flavonoids presenting intermediate
polarity. The activity of this fraction can be attributed to the presence of flavonoid aglycones commonly extracted
with ethyl acetate. The extractive process with ethyl acetate was efficient in produce fraction with high
antioxidant activity. Such activity seems to be due to non-glycoside flavonoids.
Financial Support
Coordination for the Improvement of Higher Education Personnel (CAPES); National Council for Scientific and Technological Development (CNPq), Decanato de
Pesquisa e Pós-Graduação-UnB (DPP/UnB) and Fundação de Apoio à Pesquisa do Distrito Federal (FAPDF) for financial support.
219
220
NANOEMULSION SYSTEM TO ENHANCE THE PHOTOSTABILITY OF PIPERINE
Aline Colling Schneider
Mariana Heldt Motta
Cristiane Franco Codevilla
Cristiane de Bona da Silva
Introduction Piperine is an alkaloid found in several species of the genus Piper, but in greater abundance
in Piper nigrum (black pepper). These compounds show important pharmaceutical properties such as antiinflamatory, antitumoral and antioxidant; is yellowish and sensitive towards light. Thus, it was proposed in this
work the encapsulation of piperine in a nanoemulsion system in order to protect the drug against
photodegradation. Methods Piperine nanoemulsion (P-NE) was prepared by spontaneous emulsification method.
The organic phase consisted in sorbitan monooleate, caprylic/capric triglyceride mixture, piperine (1 mg.mL-1) and
acetone, and the aqueous phase in polysorbate 80. The photodegradation assay was performed by exposing the
formulations to artificial UVA light (UVA lamp, 30 W). Samples of both the piperine solution (1 mg.mL-1) and
piperine-loaded nanoemulsion were placed individually in translucent cuvettes, at ambient temperature, and
exposed to UVA light for predetermined periods of time. The drug content was determined by the methodology
previously validated by high-performance liquid chromatography (HPLC). Results and discussion The exposition
of the piperine solution to UVA lamp demonstrated faster decrease of drug content than the piperine-loaded
nanoemulsion. After 15 minutes, the P-NE drug content remained close to 100% (1,00 mg.mL-1), while the free
piperine not associated decline to 0,90 mg.mL-1. After 4 h, the piperine solution exposed to UVA light resulted in a
drug content of 0,60 mg.mL-1, whereas in 8 h, the drug content in piperine-loaded nanoemulsion, decreased to
0,64 mg.mL-1, and in 12 h, to 0,56 mg.mL-1. These results showed the increase of stability of piperine against
UVA light. Conclusion In this study, it was demonstrated the ability of the nanoemulsion system to protect
piperine against photodegradation.
Financial Support
FAPERGS, CAPES
221
DEVELOPMENT OF LIPID-CORE NANOCAPSULES SURFACE FUNCTIONALIZED WITH
GOLD AND PHENYLALANINE
Aline de Cristo Soares Alves
Paula Sabin Raddatz
Sílvia Stanisçuaski Guterres
Introduction: Lipid-core nanocapsules (LNCs) are versatile systems widely studied with potential
biomedical applications. Bender et al. (2014) developed a new system named metal complex multi-wall
nanocapsules (MCMN). It is a system with chitosan-coated LNCs complexed with metallic ions of iron or zinc,
aiming the bond of anti-LDL(-) single-chain variable fragment. The purpose of the present study was to develop
MCMN with gold as metallic ion and phenylalanine as ligand. Phenylalanine was chosen as a model of ligand,
since it may be replaced by peptides or amino acids. Furthermore, this surface modification can contribute in the
transport across the blood-brain barrier via L-amino acid transporter i.e. LAT1. Methods: LNCs were prepared by
interfacial polymer deposition method followed by surface functionalization with ions gold and phenylalanine
constituting the MCMN nanocarrier system. This system was characterized in relation to mean and distribution
size by laser diffraction and dynamic light scattering. The zeta potential was determined by electrophoretic
mobility and particle number by nanoparticle tracking analysis (NTA). The previous physical stability of this
formulation was evaluated after 45 days of storage at room temperature and 4°C by determination of mean size
and zeta potential. Results and Discussion: The nanocapsules were successfully obtained and displayed a
volume-based mean size of 130 nm, average size of 132.28 ± 13.88 nm (n=3), polydispersity of 0.25 ± 0.05 (n=3)
and zeta potential of +13.82 ± 2 mV. The particle number found by NTA technique was of 1.53 x 10 12 / mL. The
storage at room temperature demonstrated not to be ideal, since the size and polydispersity increased to 166.80
nm and 0.43, respectively. This increase has not been observed at 4ºC. Conclusion: The developed system
showed appropriate physical-chemical characteristics indicating that the use of gold is an interesting strategy to
bind molecules on surface nanoparticles.
Financial Support
Capes / CNPq
Acknowledgments
UFRG and Capes / CNPq
222
PHENYTOIN-LOADED LIPID-CORE NANOCAPSULES: DEVELOPMENT AND
PRELIMINARY STABILITY STUDIES
Aline Marquez Cardoso
Pharmaceutical Sciences Program, Federal University of Rio Grande do Sul
Edilene Gadelha de Oliveira
Ruy Carlos Ruver Beck
Introduction: The topical therapy can be used to accelerate skin healing when structure of tissue is
injured. Nanocapsules, as delivery system to change the drug skin permeation, have been widely studied for
dermatological products. Phenytoin is an anticonvulsant drug with potential for induction of tissue healing, as an
off label use. Objective(s): The aim of this work was to develop and to characterize phenytoin-loaded lipid-core
nanocapsules for future studies regarding the skin permeation and in vivo effect. Methods: Nanocapsules were
prepared by the interfacial deposition of a preformed polymer. Their size distribution profiles were determined by
laser diffraction and dynamic light scattering. The pH of the suspensions was measured using a potentiometer.
The drug content was determined through a validated LC (liquid chromatography) method. The presence of drug
nanocrystals in the formulations was evaluated by laser diffraction and drug content (LC) during 28 days. All
samples were prepared and analyzed in triplicate. Results and Discussion: Formulations showed monomodal
particle size distribution and a mean size of 163±02 nm with a narrow particle size distribution, negative zeta
potential (−19.1±1.37 mV) and slightly acid pH (5.21±0.24). Drug content was 0.25 mg/ml, with an excellent
encapsulation efficiency of phenytoin (95±1.45%). No changes were observed during 28 days regarding the drug
content and particle size profiles. Formulations did not show the presence of drug nanocrystals. Conclusions:
Lipid-core nanocapsules containing phenytoin were effectively obtained with suitable nanotechnological properties.
They are stable at least for 28 days. Studies are in progress to produce hydrogels for skin permeation studies.
Financial Support
CNPq, CAPES and Fapergs.
Acknowledgments
Cardoso, AM thanks CAPES/Brazil for her scholarship. Authors thank CNPq, CAPES and Fapergs
for the Financial support.
223
ANTI-INFLAMMATORY AND HEALING ACTIONS OF CHITOSAN FILMS WITH
SIMVASTATIN
Laboratory of Development and Characterization of Pharmaceuticals (LDCPF). Pharmacy Department, State University of Paraíba (UEPB),
Campina Grande, PB, Brazil.
NATAN EMANUELL DE SOBRAL E SILVA
Pharmaceutical Science Graduate Program, State University of Paraiba, Campina Grande, PB, Brazil.
JOSÉ ALEXSANDRO SILVA
PEDRO GOMES SILVEIRA NETO
JOÃO WALTER DE SOUSA DA SILVEIRA
YURI BASÍLIO GOMES PATRIOTA
Pharmaceutical Science Graduate Program, State University of Paraiba, Campina Grande, PB, Brazil.
ROSEMARY SOUSA CUNHA LIMA
THARCIA KIARA BESERRA OLIVEIRA
Medical Sciences Faculty (FACISA), Campina Grande, PB, Brazil.
BOLÍVAR PONCIANO GOULART DE LIMA DAMASCENO
Chitosan (CTS) is a polysaccharide of great interest in the pharmaceutical industry primarily for
controlled release of drugs such as simvastatin (SINV). This drug is used clinically as a hypocholesterolemic
agent, but recent studies have reported significant anti-inflammatory action (pleiotropic effect of SINV). The aim of
this study was to evaluate the anti-inflammatory and healing of CTS films containing SINV (FCSINV). Four
groups of mice were analyzed utilizing film without SINV, FCSINV 20mg (3.63mg/cm2), FCSINV 10mg
(1.82mg/cm2) and saline 0.9% as control. An induced injured utilizing a punch with a diameter of 1cm was made
on the dorsal skin of mice and the films were applied immediately and replaced 2 on 2 days until euthanasia of the
animals. The analyses of the wound were made after 4, 8 and 12 days. They have been assessed by measuring
the wound area and a total number of the animal white blood cells (WBC) was performed at the end. The results
showed significant anti-inflammatory healing of 20mg FCSINV on the 8th day of onset of skin lesions, that was
observed through the reduction of wound size compared to animals treated with saline or film without SINV. A
decrease in the total number of WBC in the blood of mice was also observed in the course of time. Local
application of FCSINV in the wounds were effective in reducing the amount of WBC in all stages of lesion
evaluated. Therefore, the results demonstrate significant anti-inflammatory and healing activity of FCSINV,
favoring practical implementation of this pleiotropic effect of SINV when incorporated in chitosan films.
Financial Support
CNPq
Acknowledgments
CAPES, CNPq, FACISA.
Ethical approval
5002042015 CEUA-CESED
224
DEVELOPMENT AND ASSESSMENT OF FLOATING GASTRORETENTIVE SYSTEM
NANOPARTICLES OBTAINED BY THE EMULSION SOLVENT DIFFUSION METHOD
Ana Carolina Ferreira Pinto
University of Sorocaba
Cecília Torqueti de Barros
Katiusca da Silva Pontes
Juliana Ferreira de Souza
Marco Vinícius Chaud
Nathália Cristina Gonçalves de Rosa
Thais Francine Ribeiro Alves
Introduction: Gastroretentive systems are used for drugs better absorbed in the upper digestive tract or
to treat gastric infections, such as those caused by the Helicobacter pylori. Method: The nanoparticles were
produced by the emulsion solvent diffusion method. The process was developed in controlled conditions by using
the Easymax Basic 102 reactor. The reactional medium was stirred by using the Helix (300 rpm), magnetic (300
rpm) and Ultra-Stir (3200 rpm) type of impeller. The resulting particles of the preparing process were kept in the
reacting medium and under cooling (9°C) for 30 days. The formation of the particles and their shape and size were
observed by using optical microscopy with objective lenses of 4x, 10x, 40x and 100x magnification values. The
physical-chemical properties and the thermal-depending stability of the nanoparticles were assessed by using
zeta potential levels, diameter and polidispersity index. Results: Throughout the 30 days of storage, no
morphologic changes of the particles could be observed in function of the stirring type. The particles showed
regular size and shape, without clots. The stirring type has influenced on the zeta potential levels, diameter and
polidispersity index though. The best results were detected with the ultra-stirring type, being the zeta potential
-16,88±1,05 mV, diameter 242,77±2,68 nm and polidispersity index 0,230. After 30 days, there was an increasing
of the zeta potential levels, diameter and polidispersity index. Discussions: The analysis of the results shows that
the zeta potential levels, diameter and polidispersity index were affected by the stirring process. Conclusions: The
technique used is appropriate for obtaining nanoparticles with the potential to deliver drugs on floating
gastroretentive systems. Metronidazole is a possible drug to be used in this system as a treatment to the
infections caused by the Helicobacter pylori.
Financial Support
Finep, Fapesp
225
ANTIOXIDANT ACTIVITY AND PRESENCE OF PROBIOTIC BACTERIAS IN COSMETIC
FORMULATIONS
Ana Caroline da Costa
Paraná West State University
John Lenon Romani
Cezar Augusto Santos Souza
Fernanda Tonding Paraná
West State University
Aline Carletto
Suzana Bender
Andressa Antunes
Luciana Fariña
Introduction Photoaging process is related to the action of free radicals and the use of cosmetic
formulations containing probiotics can reduce the effects caused by this process. The aim of this study was to
evaluate the antioxidant activity of three formulations containing probiotics, two domestic and the other imported
(X, Y, Z) to determine the presence and amount of viable cells per gram of sample and identify them through their
morpho-tintorial and catalase test. Materials and Methods To evaluate the antioxidant activity was used the
method of DPPH using BHT as a positive standard. Samples were diluted and added to methanol solution of
DPPH 50mg.L-1, incubated in the dark (30 minutes) and the absorbance measured at 517nm (UV / VIS). The
antioxidant activity was calculated according to a particular formulation and expressed in %AA. To determine the
presence and quantity of viable cells, 1g of each sample was diluted in 99ml of saline containing 3% Tween 80,
used as the inactivating formulations of preservatives. To identify strains the samples were activated in TSB broth
and then centrifuged, homogenized, diluted and pour plated in series. Results and Discussion The samples had
antioxidant activity, and Z showed greater activity in the highest concentration (72%), and inferior to BHT (97%).
The X samples (64.56%) and Y (37.0%) had less activity. It was not possible to identify the presence of
probiotics, nor its amount per gram in any of the evaluated samples, indicating that the microorganisms present
were inactive. Only after activation was determined that Z formulation containing lactococcal and the others
containing lactobacilli. Conclusion: The evaluated creams showed antioxidant activity, but it was not possible to
say that this was coming exclusively from found lactobacilli and lactococcal identified.
Financial Support
CAPES; FUNDAÇÃO ARAUCÁRIA; UNIOESTE
Acknowledgments
To the financial agencies that support this research.
226
DEVELOPMENT AND CHARACTERIZATION OF CELLULOSE ACETATE
MICROPARTICLES SYNTHESIZED FROM FORAGE CACTUS (OPUNTIA FICUS-INDICA
(L) MILLER)
ANA CLÁUDIA GONÇALVES DOS SANTOS
EDUARDO BEZERRA DE ALMEIDA
ALANA RAFAELA ALBUQUERQUE BARROS
MALU MARIA LUCAS DOS REIS
MYSRAYN YARGO DE FREITAS ARAUJO REIS
FELLIPE FERNANDES SANTOS
AIRLLA LAANA DE MEDEIROS CAVALCANTI
BOLÍVAR PONCIANO GOURLART DE LIMA DAMASCENO
The modified drug release technology is one of the most promising areas of pharmaceutical science,
offering several advantages over conventional dosage forms. Polymeric microparticles have been widely used for
drug encapsulation and present increased bioavailability, less side effects and better patient compliance with
treatment. The cellulose acetate polymer is a derivative of cellulose with great commercial importance due to their
low cost, good biocompatibility and biodegradability in humans. This polymer can be extract from many plants,
among them is the forage cactus Opuntia ficus-indica (L) Miller. This study aimed the production by spray drying
and characterization of microparticulate matrix systems of cellulose acetate obtained from the cellulose extracted
from forage cactus. The cellulose acetate and the microparticles were characterized by techniques such as
Infrared Spectroscopy Fourier Transform (FTIR), Scanning Electron Microscopy (SEM) and X-ray Diffraction
(XRD). The yield of extraction of cellulose by the method adopted was 8,41%. Morphological analysis (MA)
obtained by the SEM for the cellulose acetate presents a granular appearace, material fact in defining their flow
and compression properties. To the microparticles, the MA showed like a spherical structures. The cellulose
acetate XRD showed predominantly amorphous characteristics, which is justified by increase of the degree of
hydroxyl substitution, and the microparticles have mainly amorphous regions, but they have some regions of
semi-crystallinity. FTIR showed that the cellulose acetate produced from cellulose was characteristics of a
cellulose triacetate, the microparticles for analysis proved that the methodology does not modify the polymer
profile. Thus, the obtained cellulose acetate of the forage cactus had good technological properties for application
in the production of microparticles, allowing future studies as a new alternative pharmaceutical.
Financial Support
CNPq
Acknowledgments
CAPES, CNPq, INSA
227
DEVELOPMENT AND CHARACTERIZATION OF DOXYCYCLINE HYCLATE
SUPPOSITORIES FOR VETERINARY USE
Ana Paula Christ
Pharmacy Graduate Program, Federal University of Santa Maria, RS, Brazil
Suelen Leticia Burin
Andréa Ines Horn Adams
Introduction: Doxycycline (DOX) is an antibiotic from the tetracycline class, used in human and
veterinary treatment of infections, as tablets and injection. The administration of drugs by oral route in animals is
difficult, due to the great sensibility of them. Thus, the use of suppositories can be an alternative, with the
advantage of fewer adverse effects in comparison to oral administration. The objective of this work was the
development of suppositories containing DOX to animal use. Methods: The suppositories were developed by the
molding method with different bases: hydrophilic (polyethylene glycol-PEG) and hydrophobic (cocoa butter). After
complete melting of them, doxycycline hyclate was added in order to prepare suppositories containing 50 mg of
DOX. Suppositories were characterized by mean weight, dissolution and drug content. DOX content was
determined by an electrophoretic method previously developed by our research group, by the following conditions:
a running buffer composed by solution A (sodium carbonate 25 mM and EDTA 5 mM, pH 10.6, 1:1) and B
(acetonitrile) (8:2), voltage 25 kV, temperature 24°C, detection at 260 nm using capillary with 48.5 cm (40 cm of
effective length). Dissolution test was performed using phosphate buffer pH 6.8 (500 mL), kept at 36.5ºC, and
apparatus 2, at 75 rpm. Results and discussion: Hydrophilic suppositories showed a mean weight of ± 1.18 g
(RSD=1.99%), DOX content of 99.2% and dissolution rate of 99.9% (10 min). Hydrophobic suppositories
presented mean weight of 0.88g (RSD=1.23%), DOX content of 99.4% and 79.7% of dissolution (20 min).
Conclusions: Suppositories containing 50 mg of DOX, with acceptable characteristics, were developed. Both
formulations can be considered promising for future trials. Financial support: FIT-UFSM. Acknowledgements:
Cristalia Brazil for providing doxycycline hyclate.
Financial Support
FIT-UFSM
Acknowledgments
Cristalia Brazil for providing doxycycline hyclate
228
DEVELOPMENT, STABILITY AND LEISHMANICIDAL ACTIVITY OF NANOEMULSIONS
CONTAINING ESSENTIAL OIL FOR CUTANEOUS LEISHMANIASIS TREATMENT
Ana Paula dos Santos Matos
Taisa Gomes Leiras
Vânia Emerich Bucco de Campos
Alessandra Lifsitch Viçosa
Farmanguinhos - Fundação Oswaldo Cruz
Eduardo Ricci Júnior
Carla Holandino
Introduction: Leishmaniasis belongs of the group of neglected diseases and is one of the major health
problems in the world. The current treatment for cutaneous leishmaniasis (CL) has many side effects, low patient
adhesion and parasites resistance. Nowadays, essential oils have widely application mainly their various
pharmacological effects. Therefore, this work consists in the development of nanoemulsions containing essential
oil X for CL treatment. Methods: Four nanoemulsions (N1-N4) were prepared using ultrasonication, in which
different percentages of polymer, essential oil, and surfactant were evaluated. The physical chemical properties of
nanoemulsions were characterized by droplets size distribution, polydispersity index (PdI) and pH. The
nanoemulsions were submitted to stability study during 30 days (t30), at two storage temperatures (28ºC; 4ºC).
Moreover, nanoemulsion leishmanicidal activity in L. amazonensis was investigated using resazurin assay.
Results and discussion: N1, N2 and N3 showed homogenous droplet size distribution during all the stability study
in two storage conditions (range 26.70-33.00nm, 29.50-37.60nm and 27.00-32.10nm, respectively) and lower
values of PdI (range 0.070-0.183, 0.040-0.166 and 0.060-0.234, respectively). However, N4 presented higher
droplet size (t0= 37.92±0.19, t30= 49.97±0.64 at 4ºC and t30= 39.71±0.50 at 28ºC), and higher PdI (t0=
0.151±0.00, t30= 0.363±0.01 at 4ºC and t30= 0.162± 0.01 at 28ºC). All samples presented pH=5. The resazurin
assays showed that 5µg/mL of N1-N3 were able to promote parasites death after 24 hours of incubation.
Cytotoxic assays with N4 were not conducted because of unfavorable physico-chemical results. Conclusion: Our
results indicated that is possible to obtain nanoemulsions of essential oil X with enough stability and promising
leishmanicidal potential. Additional in vitro studies are in development to confirm the nanoemulsion leishmanicidal
activity. Financial support: CAPES.
Financial Support
CAPES
Acknowledgments
IMA-UFRJ
229
EVALUATION OF THE CHEMICAL, PHYSICAL AND MICROBIOLOGICAL STABILITY OF
NANOEMULSIFIED SYSTEMS CONTAINING EXTRACT OF FRUITS OF RAPANEA
FERRUGINEA
Angélica Garcia Couto
Odair José Custodio Junior
Ana Cristina Gon
Josiane de Carvalho Vitorino
Mariana Cristina Cechetto
Tania Mari Belle Bresolim
Angélica Garcia Couto
Introduction: The ethanolic extract of Rapanea ferruginea fruits has shown promising analgesic and antiinflammatory effects, when incorporated in nanoemulsions. This study aimed to evaluate the stability of a
nanodispersed oil in water (o/w) system containing 1% of R. ferruginea fruits extract (NRF). Methods: Both NRF
and vehicle were prepared by phase inversion method with stirring (600 rpm) under heating, using 10% of
surfactant (Ultramona RH 400® and Span®), 5% of oil (Polymol®) and 1% of preservative (Phenonip®). The
nanoemulsions were storage in amber glass bottles and submitted to accelerated stability study at 40 ºC, 30 ºC
and 5 ºC ± 2 ºC, for 60 days with sampling at 0 (24 h), 45 and 60 days. The systems were characterized in
triplicate by the mean droplets size, polidispersibility index (PDI), zeta potential, measured in Malvern Zetasizer®
and pH. The mirsinoic acid A (AMA) content was used as a chemical marker, and quantified by HPLC validated
method. Results and Discussion: The pH of NRF was slightly more acid (pH = 5.8) than vehicle (pH = 6.1),
keeping relatively constant over the study. The mean droplets size was similar (29.86 ± 3.59 nm) to vehicle (34.98
± 7.31 nm, but with higher PDI (0.476 ± 0.16) than vehicle (PDI = 0.196 ± 0.05). However, NRF had a zeta
potential of -36.85 ± 7.85, against -11.05 ± 1.34 of vehicle. Indeed, the NRF showed to be much more stable than
vehicle with minor variations in mean droplet size, PDI and zeta potential. The microbiological analysis of NRF
showed less than < 102 CFU/g for bacteria, fungi and yeast and absence of pathogenic bacteria, in accordance
with acceptance criteria of Brazilian pharmacopeia, The NRF was characterized with AMA content of 0.744 ±
0.017 mg/ml after 24 h, keeping relatively constant over 60 days at different temperatures of storage. Conclusion:
NRF showed a good stability with variations between 5% and 10% over 60 days, with promising characteristics
for future clinic trials.
Financial Support
Article 170 from State Government of Santa Catarina; FAPESC (Fundação de Amparo à Pesquisa e Inovação do Estado de Santa Catarina), Edital PRONEM
(06/2012) FAPESC/CNPq, grant number 2708/2012.
Acknowledgments
Angela Malheiros and Taylin Zermiani for providing the herbal drug and chemical marker
230
DEVELOPMENT OF PLGA NANOPARTICLES: A COMPARATION OF TWO METHODS
OF PRODUCTION
Anna Emmanuela Medeiros de Brito
Alexsandra Conceição Apolinário
Universidade de São Paulo (USP)
Thayse Silva Medeiros
Mysrayan Yargo de Freitas Araujo Reis
Malu Maria Lucas dos Reis
Carlota de Oliveira Rangel Yagui
Adalberto Pessoa Junior
José Alexsandro da Silva
Introduction: Polymeric nanoparticles (NP) are largely produced by the double emulsification method and
poly (lactic-co-glycolic acid) (PLGA) is one of the most used polymers, due its biodegradable and biocompatible.
In this study, we investigated the preparation and characterization of PLGA-NP to future application for Lasparaginase protein encapsulation. Methods: Two different molecular weight (MW) of PLGA polymers were
used: A-MW = 30-60 kDa and B-MW = 24-38 kDa. PLGA (0.05 g) was diluted in chloroform (2.5 ml) and
dispersed in 250 µl Milli-QTM water followed by sonication over an ice bath for three cycles of 60 s. This first
emulsion was added into 10 ml of 1% polyvinyl alcohol solution (PVA) and it was repeated sonication cycle. The
solvent was evaporated (24 hours) under reduced pressure and the emulsions were divided into two parts: 1) noncentrifuged; 2) submitted to two centrifugation steps at 4000 rpm for 15 minutes at 25 °C and resuspended with 2
ml of water. The hydrodynamic diameter of the PLGA-NP was messure by dynamic light scattering (DLS)
technique before and after lyofilization. Results and Discussion: The DLS measurement yielded hydrodynamic
diameters of 565.4 nm and 847.6 nm for NP of PLGA-A non-centrifuged and centrifuged, respectively. For the
system formed by NP of PLGA-B, the hydrodynamic radius was 465.5 nm and 747.7 nm for system noncentrifuged and centrifuged, respectively. Therefore, it can infer that the spin may result in aggregation of NP,
what result in increasing the apparent hydrodynamic radius. All NP in this study were homogenous with
acceptable values of polydispersity index (<0.3), and there was no significant variation in size between the
lyophilized and non-lyophilized systems. Conclusion: It can be concluded that NP obtained with PLGA-B were
smaller and less polydisperse and the centrifugation step interfered in the stability of preparations potentiating
their aggregation.
Financial Support
CNPq, FAPESP, UEPB
231
HOSTGUEST SYSTEM OF CARVONE AND βCYCLODEXTRIN: PREPARATION AND
CHARACTERIZATION
Antonio Guilherme de Carvalho Neto
Departament of Pharmacy, Federal University of Sergipe, São Cristóvão, SE, Brazil
Yasmim Maria Barbosa Gomes de Carvalho
Bruno dos Santos Lima
Gabriela das Graças Gomes Trindade
Paula dos Passos Menezes
Bruna Maria Hipólito de Sousa
Dayanne Valéria Soares Santana
Lucindo José Quintans Júnior
Departament of Physiology, Federal University of Sergipe, São Cristóvão, SE, Brazil
Adriano Antunes de Souza Araújo
Mairim Russo Serafini
Introduction: βcyclodextrin (βCD) is a cyclic oligosaccharide that present ability to form non-covalent
inclusion complexes with lipophilic molecules such as carvone (CA), monoterpene, unsaturated, slightly soluble in
water, have the properties of anti-inflammatory, antinociceptive, anticonvulsant and anxiolytic. Methods:
Complexes were obtained by physical mixture (PM), kneading (KD), slurry complex (SC) and freeze drying (FD)
were investigated by differential scanning calorimetry (DSC), thermogravimetry/derivative thermogravimetry
(TG/DTG), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), X-ray diffraction
(XRD) and entrapment efficiency (EE). Results and Discussion: DSC curves corresponding of KD, SC and FD
showed disappearance of event characteristic CA's volatilization (93.8 °C). TG curves of FD showed mass loss
(5.18%) between 34-136 ºC, indicating the substitution of water molecules cavity βCD by molecules of the CA.
FTIR results of KD, SC and FD presented characteristic bands (1680 cm-1) CA's ketone group. XDR analysis
the βCD exhibited crystalline peaks between 3° and 40° (2θ = 4.48, 8.87, 10.55, 12.46 and 15.41°) confirming its
crystalline form. KD, SC, FD's diffractogram exhibited the disappearance βCD's spectral lines at 4.48° and
15.41° (2θ), new peaks of emergence 5.92° and 7.07° (2θ) was observed, suggests the formation of inclusion
complex. SEM results showed for βCD crystalline structure and FD decrease size and structure crystalline loss.
The EE results showed that FD method was the best, because presented EE 84,26%, confirming the formation of
inclusion complex. Conclusions: CA/βCD complexes are considered as a promising carrier system to preserve
volatile and hydrophobic drugs enlarging their application in cosmetic and pharmaceutical industries.
Financial Support
FAPITEC/SE
Acknowledgments
CAPES, CNPq, FINEP, FAPITEC/SE
232
NANOCAPSULES SUSPENSIONS DEVELOPMENT CONTAINING Psidium cattleinum
Sabine ESSENTIAL OIL
Ariane Schiavenin
Fabiana Agostini
Melissa Schwanz
Ronaldo Adelfo Wasumᵻ
Valéria Weiss Angeli
Barbára de Antoni Zoppas
Introduction: Psidium cattleinum Sabine (Myrtaceae) among all its properties the essential oil (EO)
application against fungal infections is highlighted. The EO is unstable therefore measures to increase the
chemical stability are required. The aim of this work is the obtainment and characterization of the EO, evaluation
of its antifungal capacity and the association of the EO in polymeric nanocapsules (NC). Methods: The oil was
obtained by hydrodistillation and the characterization was conducted by HP®6890GC/MS. Only the oil antifungal
activity was determined in this preliminary study using the diffusion method and C. albicans strain. Medium
Sabouraud Dextrose Agar solidification, the culture of C. albicans was seeded and 100µL of EO and 100 µL of
0.3% solution nystatin were applied on cannulas. The samples were incubated (37ºC ± 2ºC/24-48 hours). The
inhibition zone was determined by measuring the clearance zone diameter. NC were prepared by the interfacial
deposition of the preformed polymer method. Poly (Ɛcaprolactone), sorbitan monoestearate, and the EO were
dissolved in acetone. The polysorbate 80 was dissolved in water. The organic solution was added into of aqueous
phase under magnetic stirring. Acetone was evaporated and the suspension concentrated under reduced pressure
(10mg/mL of EO). The NC were pH based on pH meter Digimed®. The zeta potential, z-average and
polydispersity index (PI) were based on Microtrac® Zeta Sizer Nano. Results and Discussion: The major oil
compound found was 1,8-cineol (63.41%). Antifungal oil essays showed an inhibition halo of 15mm, against
12mm for nystatin. NC presenting pH value 6.18±0.02, z-average 314±37.36 nm diameters, zeta potential -35 mV
and PI higher than 0.25. Conclusions: It was possible to obtain and perform the chemical EO characterization,
and also to evaluate its antifungal properties. Nanoencapsulation of EO seems to be promising; however, it is a
preliminary study requiring further testing.
Acknowledgments
Universidade de Caxias do Sul
233
IN VITRO RELEASE OF COUMESTROL/HYDROXYPROPYLβCYCLODEXTRIN
ASSOCIATION FROM NONIONIC AND CATIONIC HYDROGELS
Barbara Elisabeth Kummer Machado
Sara Elis Bianchi
Cristiana Lima Dora
Fundação Universidade Federal do Rio Grande (FURG)
Valquíria Linck Bassani
Introduction: Coumestrol is found in many Fabaceae species which presents antioxidant and estrogenic
activity. However, its use in pharmaceutical and cosmetic formulations is limited by its low aqueous solubility,
hence the interest of its association with cyclodextrins. The aim of the present study is to compare two different
methods of association of coumestrol with hydroxypropylβcyclodextrin (HPβCD), to incorporate the association
into nonionic (HPMC) and cationic (chitosan) hydrogels and evaluate the coumestrol in vitro release in comparison
to free coumestrol. Methods: Two methods of coumestrol:HPβCD association were evaluated, as follows:
microwave method followed by freeze drying (MWL), organo-aqueous method followed by spray drying (OASD).
Physical mixturewas prepared in a mortar. The associations and free coumestrol were incorporated into hydrogels
in a mortar and in vitro release tests were performed using Franz type cells. Results and Discussion: The
association coumestrol:HPβCD obtained by MWL method resulted in a heterogeneous powder instead that
obtained by OASD. The last one resulted in spherical and homogeneous particle being selected to be incorporated
in the hydrogels. The coumestrol release from hydrogels containing the physical mixture shows similar release
profile than free coumestrol (0,1%). HPβCD increased the coumestrol release from both hydrogels reached 100%
in 12 hours. Conclusions: The association coumestrol/HPβCD obtained using OASD presented better
technological caracteristics to be incorporated into hydrogels. HPβCD revealed clear and significant promotion of
the release of coumestrol from both hydrophilic matrices, nonionic (HPMC hydrogel) and in cationic (chitosan
hydrogel).
Financial Support
Financial support from Brazilian government (CNPq) and master scholarship from CAPES.
234
IN VITRO RELEASE PROFILE AND ANTI-INFLAMMATORY ACTIVITY OF
PENTOXIFYLLINE-LOADED MICROEMULSION
Mysrayn Yargo de Freitas Araújo Reis
Geilza Carla de Lima Silva
Karina Lidianne Alcântara Saraiva
Pentoxifylline (PTX) is involved in modulation of immunological processes as inflammatory skin
diseases. Conventional oral dosage form of this drug often fails to achieve suitable concentrations in the skin to
generate maximum pharmacological effect. Incorporation of PTX into microemulsion (ME), could allow the
optimization of aspects such as bioavailability and efficacy in therapy. This study aimed to develop PTX-ME for
topical use and evaluate its in vitro release profile and in vivo antiedematogenic activity. The formulation was
selected based on pseudo-ternary phase diagram. The in vitro release kinetics assays from PTX-ME was
determined using the model of Franz cells. And the in vivo antiedematogenic activity was determined using the
technique of paw edema induced by carrageenan, in which PTX solutions (36 and 72 mg/kg) were injected
intraperitoneally and the treatment with the blank-ME and PTX-ME (7.2 and 14.4 mg/kg) were done by topical via.
ME selected was composed of distilled water, acid capric/caprylic triglycerides, a mixture of TweenTM 80 and
BrijTM 52 and PTX 1%. The amount of PTX in vitro released from ME was 66% and followed the Higuchi kinetic
model suggesting that the PTX diffusion mechanism through the vehicle was the limiting step in the release.
Steady state flux was 582.37 µg/cm2 and the PTX Iag time from the ME was 3.18 min. These values were
promising to develop a permeation study. PTX-ME topical application showed significant anti-inflammatory activity
in paw edema in all stages of the assay and it was able to develop a superior activity when compared to the PTX
solution reducing up to 88% the paw edema. Therefore, these data suggested that PTX-ME is a promising
alternative for PTX topical use and a potential therapeutic agent for skin diseases.
Financial Support
CAPES and CNPq.
Acknowledgments
CETENE and CERTIBIO.
Ethical approval
0038/18122013
235
DEVELOPMENT OF INDOMETHACIN ALGINATE MICROPARTICLES
Joilly Nilce Santana Gomes
Kevin da Silva Oliveira
Rosemary Sousa Cunha Lima
Joandra Maísa da Silva Leite
Ana Cláudia Gonçalves dos Santos
Bolivar Ponciano Goulart de Lima Damasceno
The development of indomethacin (IND)-alginate microparticles as an IND controlled release system
can be an alternative to reduce gastric irritation and ulcerative lesions caused by chronic use of this antiinflammatory. This study aimed to produce alginate microparticles with and without IND and evaluate their yield
and morphology. The microparticles were produced in two alginate concentrations, M1 (1.5%) and M2 (2%), with
300mg of IND. After homogenizing the solution by magnetic stirring, the samples were nebulized using a spray
dryer, following standardized conditions (0.3 L/h, 40 L/min; 75°C; 3.5 m³/min). The product was weighed before
and after atomization to determine the yield and the morphology of samples were analyzed by fluorescence
microscopy. The yield of M2 was higher than M1, 39±0.5 and 35±0.5%, respectively, and for IND microparticles
there was an increase in the yield to both M2 and M1, 49±0.5% 47±0.5%, respectively. By naked eyes, the
samples with IND presented more yellowish. The microscopy analysis of microparticles show them deformed and
with presence of agglomerates. The M1 samples with and without IND presented rougher and less spherical than
M2. Beside of this, M2 has greater presence of agglomerates. Therefore, it can be seen that the samples with
higher polymer concentration have become more feasible, because it has a higher yield and more spherical and
less rough form, which require further optimization of the method for standardization of microparticles proposed in
the study.
Financial Support
Cetene, Certbio
Acknowledgments
Capes, Cnpq
236
INVESTIGATION OF MYRTENOL INCLUSION COMPLEXES WITH BETACYCLODEXTRIN: DEVELOPMENT AND CHARACTERIZATION
Bruno dos Santos Lima
Dayanne Valéria Soares Santana
Department of Pharmacy, Federal University of Sergipe, São Cristóvão, SE, Brazil
Isabella Gonçalves Matos
Antonio Guilherme de Carvalho Neto
Valléria Matos Andrade
Lucindo Jose Quintans Júnior
Department of Physiology, Federal University of Sergipe, São Cristóvão, SE, Brazil
Introduction: (-)-Myrtenol is an alcoholic monoterpene is used to treat stomach problems, insomnia and
nervous disorders. This compound has low solubility in water. Hence, the present study aimed to prepare the
favorable complexation of this compound with beta-cyclodextrin (beta-CD). Methods: The complexes were
prepared by physical mixture (PM), paste complex (PC) and slurry complex (SC) method and the sample was
characterized using differential scanning calorimetry (DSC), thermogravimetry/derivative thermogravimetry
(TG/DTG), X-ray diffraction (XRD) and scanning electron microscopy (SEM) methods. Results and Discussion:
The myrtenol DSC curves showed endothermic events between 200-240ºC relative the volatilization and beta-CD
curves demonstrated three endothermic events. The first (30-113ºC) one is correspondent of the water release,
the second one is (210-230ºC) characteristic of the crystalline phase transition and the third is (280-340 ºC)
attributed the fusion. The PCand SC DSC curves exhibited the disappearance of crystalline phase transition and
a reduction in the melting peak temperature of beta-CD. The PC and SC TG/DTG curves showed weight loss
decrease (11,68% and 14,25%) between 30-170ºC and increase (4,55% and 3,88%) between 170-280ºC,
respectively. This happens, because of the complexation process some of water molecules are replaced by
hydrophobic molecules. The SC XRD results reveale the decreasing peak intensity characteristics of crystallinity,
suggesting amorphization of beta-CD crystals for the interaction myrtenol/beta-CD, corroborating with the SEM
results that showed particle size reduction and alteration in rectangular-shaped crystals. Conclusions: According
to the present results, suggested that the inclusion complexes formation between myrtenol/beta-CD for the SC
method is the best of complexation.
Financial Support
CAPES, CNPq, FINEP and FAPITEC/SE
237
PREPARATION OF DRIED PRODUCTS FROM CHRYSIN-LOADED COLLOIDAL
SUSPENSIONS
Camila Reck Rampelotto
Alessandra Scherer Lorenzoni
Gabriele dos Santos Depieri
Sabrina Negrini Bolson
Milena Libardoni
Scheila Rezende Schaffazick
Introduction. Chrysin is a natural compound and possesses antioxidant and antitumor activities.
However, this flavonoid has poor solubility in water and its bioavailability in the body is low. Polymeric
nanocapsules can be alternative systems to delivery of this substance. Hence, the aim of the present study was
to prepare freeze-dried chrysin-loaded nanocapsules, using ethylcellulose and medium chain triglycerides, in order
to obtaining stable solid dosage forms. Methods. Nanocapsule suspensions were prepared by interfacial
deposition of preformed polymer. The nanocapsule suspensions containing chrysin were frozen and lyophilized
(LIOTOP-L101, Liobras), using trehalose (10% w/v) as cryoprotectant. The nanocapsule suspensions were
evaluated for their mean particle sizes, polydispersity indexes (PdI) and zeta potentials (Zetasizer® NanoseriesMalvern). The encapsulation efficiency was also performed (Amicon®). The lyophilized products were analyzed
by scanning electron microscopy (SEM) and dynamic light scattering (Zetasizer®) after redispersion in water and
filtration of the samples. The stability of lyophilized products was also evaluated (HPLC; 30 days/room
temperature; protection from light). Results and Discussion. The chrysin-loaded nanocapsule suspensions
presented average diameter lower than 120 nm, adequate PdI (<0.2) and negative zeta potential. Besides, high
encapsulation efficiency was verified. After preparation, the chrysin content of dried product was close to the
theoretical value. Analysis by SEM showed the presence of colloidal spherical particles in freeze-dried samples.
After redispersion in water, the mean particle sizes of dried products were similar to the sizes of the original
suspensions (p>0.05). The dried formulations showed stable chrysin content during 30 days of the study (p>0.05).
Conclusion. It was possible to obtain redispersible products with suitable characteristics by freeze-drying of
chrysin-loaded nanocapsule suspensions.
Financial Support
CAPES; FIPE Sênior/UFSM program
238
SUPERCRITICAL CARBON DIOXIDE FOR POLYMER PROCESSING AND SCAFFOLD
MANUFACTURING
Cecília Torqueti de Barros
Pharmacy Graduate Program, University of Sorocaba, Sorocaba, SP, Brazil; Laboratory of Biomaterials and Nanotechnology. University of
Sorocaba. Sorocaba, SP. Brazil.
Katiusca Da Silva Pontes
Sorocaba. Sorocaba, SP. Brazil
Sorocaba. Sorocaba, SP. Brazil
Laboratory of Biomaterials and Nanotechnology. University of Sorocaba. Sorocaba, SP. Brazil
Nathalia Cristina Gonçalves de Rosa
Marco Vinicius Chaud
Introduction: Supercritical carbon dioxide (ScCO2) processing may be used to form foamed scaffold
(SCF). These processing methods possess advantage over standard processing methods for the production of
SCF for use in tissue engineering. The process allows control over. ScCO2 processing allows control over the
morphology and the architecture of the scaffold pores. Simultaneous change in phase in the CO2 and polymers
results in rapid expansion of surface area and changes in the polymers properties. Objective: The aim of this
study was obtained SCF by ScCO2 processing. Methodology: Processing parameters under investigation were
temperature and pressure. The Supercritical Anti Solvent (SAS) method was utilized as supercritical processing.
A factorial design with duplication of central point (32 + 2) was utilized for altered the supercritical system
parameters. The PLLA polymer was used dissolved in dichloromethane (DMC) and sprayed to the inside of a
chamber precipitation of the device at a controlled rate. The structure and porosity of SCF was characterized by
microtomography computed (µTC). Results and Discussion: The scaffolds were obtained with 98% porosity and
80% interconnectivity. The average volume of the pores in the scaffold was 31.17 ± 5.60mm3. CO2 from
plasticized polymer melt generates gas bubbles that shape the developing process. Higher pressures followed by
a long immersion time leads to a higher nucleation density and pore formation. Higher temperatures facilitate the
diffusion and increase the porosity. The reduction in the depressurization rate increases the amount and volume of
pores. Conclusion: This study has demonstrated that the pore size, volume and structure of the supercritical
PLLA scaffolds can be tailored by careful control of ScCO2 processing conditions.
Financial Support
CNPq, FAPESP. FINEP. FAPIC - UNISO
239
EXPERIMENTAL DESIGN TOWARDS AN OPTIMAL LIPID NANOCARRIER FOR
TUBERCULOSIS TREATMENT
Cristiana Lima Dora
Universidade Federal de Rio Grande
PRISCILA C. B. HALICKI
UNIVERSIDADE FEDERAL DE RIO GRANDE
GABRIELA HADRICH
DANIELA F. RAMOS
L ANDRADE
RAPHAEL BOSCHERO
JULIANA BIDONE
HELDER FERREIRA TEIXEIRA
ANA LUIZA MUCCILLO-BAISCH
PEDRO E. A. SILVA
Introduction: Tuberculosis (TB) is an infectious disease that is still considered a global public health
emergency. The long duration of the treatment combined with toxic effects can hamper patient life-style and
induces patient non-compliance or microbial resistance. Strategies using nanotechnology represent a new tool for
the TB treatment. Additionally, many excipients have pharmacological activities and exploring their history of safe
use in therapeutics is a smart strategy to obtain improved pharmaceutical products. This study aims to evaluate
the anti-TB activity of some excipients to further design an optimal lipid nanocarrier for tuberculosis treatment.
Methods: Castor oil (CO), PEG 660-stearate and soy lecithin was choosing as excipients. The excipients were
first solubilized in proper diluent following the particular chemical characteristic of each one. All this components
were evaluated against one multidrug resistant clinical isolate of M. tuberculosis (Strain 1), with mutations in katG
(S315T) and rpoB (S531L) genes, and a pan-susceptible strain H37Rv (ATCC 27294), by Resazurin Microtiter
Assay method. Results and Discussion: The minimum inhibitory concentration (MIC) of CO and PEG 660stearate was, respectively, 117.2 and 468.8 µg/ml to strain 1 and H37Rv. Soy lecithin showed no activity against
M. tuberculosis strains up to a concentration of 250 µg/ml. The antimycobacterial potential in vitro suggests that
castor oil and PEG 660-stearate are important candidates to design a lipid nanocarrier for TB treatment since they
have similar activities for two strains with different susceptibility profiles. Conclusions: Castor oil and PEG 660stearate are interesting excipients to be used to design a drug delivery system for the treatment of Tuberculosis.
Financial Support
National Council for Technological and Scientific Development (CNPq) Casadinho/Procad 552457/2011-6 and CAPES
240
DEVELOPMENT OF MOUTHWASH CONTAINING CETYLPYRIDINIUM CHLORIDE FOR
CANCER PATIENTS.
Cristiane Del Bello
Marina Goulart da Silva
Ana Julia Pereira Santinho Gomes
Introduction. Basic oral care is considered a mainstay of supportive care in cancer patients, since it
may decrease noninfectious oral complications (mucositis and bleeding). It is known that cetylpyridinium chloride
is a quaternary ammonium cationic surfactant used as an antiseptic agent for oral and throat care due to inhibit
plaque formation. It reduces the influence of oral bacterial flora, symptoms of pain and bleeding related to anticancer therapy besides to preventing infections in tissue. Nonetheless, available mouthwashes have caused pain,
burning and oral discomfort for patients undergoing radio- or chemotherapy. Objective. To develop and evaluate a
mouthwash free of substances with hydroxylic structures (Ethanol, Glycerin, Menthol, Propylene Glycol and
Sorbitol), adjusting the composition for management of oral mucositis in cancer patients. Methods. After
organoleptic and physicochemical comparisons with others mouthwashes, it was prepared a formulation (n=3)
containing cetylpyridinium chloride, polysorbate 20, sucralose, sodium hydroxide (pH 6.5±0.5), soothing flavor and
water. Samples were submitted monthly to determinations of pH, specific gravity and viscosity. Based on an
analytical curve, determination of cetylpyridinium chloride in solution was carried out at 259 nm using acid water.
Results and discussion. Prepared formulation maintained its initial properties such as appropriate pH value
(6.6±0.20), specific gravity (1.001±0.001 g.cm-3) and very low viscosity. Spectrophotometric method showed that
solution contained not less than 95% and not more than 105% of the labeled amount of cetylpyridinium chloride.
Conclusion. It is suggested that the prepared formulation presents suitable properties to be used uninterruptedly
as mouthwash between others basic oral care products for the management of oral mucositis in cancer patients,
since the degree of mucositis can be crucial to the continuation of the therapy or if it will need to be stopped.
Acknowledgments
UFSJ
241
MECHANISMS UNDERLYING IN VITRO ANTINEOPLASTIC ACTIVITY OF
DOXORUBICIN-LOADED pH-RESPONSIVE NANOPARTICLES
Daniele Rubert Nogueira Librelotto
Francieli Bellé
Universitat de Barcelona
Introduction: Stimuli-responsive delivery systems, in particular those that are sensitive to pH variations,
might allow the exploitation of the various pH gradients within the body, i.e. between healthy and tumor tissue, or
between the extracellular tissue and some cell compartments. Doxorubicin (DOX) is a hydrophilic
chemotherapeutic drug that binds to DNA by intercalation, leading to apoptosis in tumor cells. Here, our study
focuses on a strategy to improve the efficacy of DOX assisted chemotherapy, by developing pH-responsive DOXloaded chitosan'tripolyphosphate nanoparticles (CS-NPs). Methods: CS-NPs with pH-responsive behavior were
obtained by ionotropic gelation method, using the surfactant 77KS, derived from the amino acid lysine, as a pHsensitive adjuvant. HeLa and MCF-7 tumor cells were used to assess the in vitro antiproliferative activity of either
CS-NPs or free drug. Two different endpoints (MTT and NRU assays) were applied to monitor the viability
responses. The mechanisms underlying the cytotoxic responses were studied by means of apoptosis and cell
cycle experiments, using flow cytometry. Finally, the cellular uptake of CS-NPs was also assessed by flow
cytometry. Results and Discussion: The in vitro antiproliferative assays indicated that the NPs themselves had
no associated significant cytotoxicity, while DOX-loaded pH-responsive NPs displayed higher cytotoxicity than
free drug. Flow cytometry analysis evidenced that CS-NPs were internalized by HeLa cells. Likewise, it was
evidenced that the combined physicochemical and pH-responsive properties of CS-NPs allowed the entrapped
drug to induce greater cell cycle arrest and apoptotic effects. Conclusions: The insights into some cytotoxicity
mechanisms of DOX-loaded CS-NPs contribute to understanding the behavior underlying their antiproliferative
activity. Moreover, the overall results suggest that these pH-responsive NPs could be potentially useful as a drug
delivery platform for cancer therapy.
Financial Support
Grants 447548/2014-0 and 401069/2014-1 of CNPq and 2293-2551/14-0 of FAPERGS.
Acknowledgments
Nogueira-Librelotto DR and Scheeren LE thank CNPq and FAPERGS for the Postdoctoral and Masters¿ fellowships, respectively.
242
Evaluation of Rheological Properties and antifungal activity of the Sapindus saponaria
L. extract on formulation for the treatment of Vulvovaginal Candidiasis
Danielle Rodrigues de Souza
State University of Maringá
Enikeyla Azevedo Sudati
Bruna Luiza Pelegrini
Janine Pinheiro Marques
Thaisa Yumi Takeda Violin
Marli Miriam de Souza Lima
Izabel Cristina Piloto Ferreira
Introduction: Vulvovaginal candidiasis is a fungal infection of vulva and vagina, caused by several
species of Candida, mainly by C. albicans. The conventional treatment is accomplished through the
administration of imidazole and triazole compounds, and polyenic agents, leading to serious problems of
intolerance and toxicity in the gastrointestinal tract. Recently, many studies using plants has been carried out in
search of new antifungal agents, currently by the localized action of the drug. Methods: In this study, a gel using
carboxivinilic acid polymers (Carbopol®) at concentrations of 0.5 and 0.7% (w/w) containing 5% (w/w) of extract
of pericarp of Sapindus saponaria L. was developed for the treatment of vulvovaginal candidiasis. Fruits of S.
saponaria L. was collected in the State University of Maringá. The rheological properties were carried out in
rheometer MAARS-HAKE with cone plate geometry and spindle C35/2º-Ti. Results and Discussion: All
formulations exhibited non-Newtonian, pseudoplastic and thixotropic characteristics and there
was
a
predominance of elastic character, which resulted in a gel behavior. Antifungal activity were characterized and the
two vaginal formulations were evaluated against two strains of Candida albicans, one C. glabrata, one C.
tropicalis and two strains of C. albicans ATCC 90028 and C. parapsilosis ATCC 22019. Both formulations
showed antifungal activity with minimum inhibitory concentrations ranging from 780 to 1.560 µg/mL and the
minimum fungicidal 780 to 3.125 µg/mL. Conclusion: According to the results, vaginal gel of Carbopol® containing
S. saponaria extract in 5% becomes a therapeutic alternative for the treatment of vulvovaginal candidiasis.
Financial Support
CAPES
Acknowledgments
CAPES
243
Development and characterization of methotrexate-loaded functionalized PLA
nanoparticles
Deise Lidiane de Sousa
Iaponira Roque Barboza
Emanuell dos Santos Silva
Renata de carvalho Feitosa
Alaine Maria dos santos Silva
Lília Basílio de Caland
Arnóbio Antônio da Silva Júnior
Introduction: Methotrexate (MTX) is an antimetabolite drug used in the chemotherapy of cancer,
autoimmune diseases and uveits. Their pharmacokinetic limitations include short half-life time and difficulty to
across biological barriers. Cationic nanoparticles (NP) can improve the drug efficacy due to the ability to
overcome biological barriers reaching specific tissues. This study focused in the preparation and optimization of
cationic and fluorescent functionalized MTX-loaded polylactide (PLA) NP by the emulsification with solvent
evaporation method. Methods: The effect of different surfactants (polysorbate 80, sorbitan monooleate 80,
polyvinyl alcohol (PVA), Poloxamer 188 and Poloxamer 407), as well as distinct surfactant/polymer ratios were
tested. The particle size, zeta potential, Atomic Force Microscopy (AFM) images and encapsulation efficiency
were used as performance parameters to select the best formulation. The selected formulation was functionalized
with polyethyleneimine (PEI 25 kDa) in PEI:PLA proportion of 1:1 and fluorescent labeled with fluorescein
isothiocyanate (FITC). Results and Discussion: MTX-loaded nanoparticles exhibited size of 260.53 nm ± 1.80 and
zeta potential of +34.13 mV ± 2.13, with encapsulation efficiency of 83.89% ± 3.56. AFM images demonstrated
spherical and smooth particles, which have the fluorescence assessed by fluorescence microscopy images,
Fourier transforms Infrared (FTIR)-ATR and flow cytometry. Conclusions: Thus, cationic and fluorescent
functionalized MTX-loaded PLA nanoparticles were successfully optimized by using emulsification with solvent
evaporation method.
Financial Support
Capes PNPD (23038.007487/2011-91) and CNPq (481767/2012-6)
Acknowledgments
Capes/Brazil (Sousa DL, Barboza IR, dos Santos-Silva E, Silva AMS, Caland LB,
244
DEVELOPMENT OF SPRAY-DRIED PHENYTOIN-LOADED CATIONIC NANOCAPSULES
Edilene Gadelha de Oliveira
Aline Marquez Cardoso
Introduction: Nanocapsules have been studied for brain delivery of drugs. However, they can present
limited physical stability during storage. Spray-drying has been widely used to produce dried nanocapsules. In this
study, spray-dried cationic nanocapsules containing phenytoin were developed for further in vitro and in vivo
studies. Methods: Cationic nanocapsules were produced by the interfacial deposition method, using poly(ε
caprolactone), as polymer. Cationic coating was done using a chitosan solution at 0.6%. They were characterized
according to the following properties: particle size, zeta potential, pH and drug content. Nanocapsules were spraydried in Mini-Spray Dryer (Büchi, B-290), using these operational conditions (Inlet temperature, 120°C; outlet
temperature, 68°C; air flow rate, 600 NL/h; aspirator, 100% and feed rate 4.5 ml/min). Maltodextrin was used as
drying adjuvant at 10% (w/v). The powders were characterized by yield, loss of drying, particle size distribution,
and morphological analysis. Results and Discussion: Phenytoin-loaded nanocapsules had mean particle size of
168 nm ± 04, zeta potential of +16 mV ± 1.51 and pH of 4.00 ± 0.08. These properties were not influenced by the
presence of the drug. Drug content was 0.25 mg/ml and encapsulation efficiency of 93%. They were stable for 14
days at room temperature. Spray-dried powders presented high yields (60-80%) and low loss of drying (up to 4%).
Moreover, SEM images showed nanostructures on the surface of the microparticles. After aqueous dispersion on
a silicon wafer, nanocapsules were visualized. It can also be observed by size particle distribution, demonstrating
a good aqueous redispersion. Conclusions: The powders containing phenytoin-loaded cationic nanocapsules
showed suitable properties regarding the production process and good deagglomeration profile, which is essential
for oral redispersible powders. Further studies will be carry out to evaluate their in vitro and vivo performance.
Financial Support
CNPq/Brazil, FAPERGS and CAPES.
Acknowledgments
Thanks CAPES/Brazil for the PhD scholarship.
245
PREPARATION AND PHYSICOCHEMICAL CHARACTERIZATION OF METHOTREXATELOADED CATIONIC FUNCTIONALIZED PLGA NANOPARTICLES
Renata de Carvalho Feitosa
Alaine Maria dos Santos Silva
Lília Basílio de Calland
Introduction: Methotrexate (MTX) is an antimetabolite commonly used in the treatment of cancer and
autoimmune diseases. However, its pharmacokinetic characteristics limitations such as poor absorption and short
half-life time limits the therapeutic efficacy of drug. Polymeric nanoparticles (NP) are used as drug delivery
system for improving the solubility, stability and bioavailability of drugs, especially the cationic NPs that easily
overcomes some biological barriers. This study focused in the preparation small sized cationic functionalized
MTX-loaded poly (lactic-co-glycolic acid) (PLGA) nanoparticles by using nanoprecipitation method. Method: The
polymer was added into the acetone organic phase, which was injected into the aqueous phase containing the
surfactant (polaxamer 407 or 188) in different concentrations. Different PLGA:polyethylenimine (PEI 25 kDa )
ratios (1:7, 1:10, 1:14) were tested to obtain cationic particles. The drug loading efficiency of distinct formulations
was tested for MTX:PLGA ratio of 1:10 (w/w). Size and zeta potential were monitored by Dinamic Light
Scattering, while the shape and surface of NPs were assessed by using Atomic Force Microscopy (AFM)
images. Results and Discussion: Polaxamer 407 exhibited best performance as surfactant, producing particles in
the range of 99 to 146nm. The formulation containing 0.5% of polaxamer 407, 0.75% of PLGA and PEI: PLGA
ratio of 1:10 exhibited narrowed sized particles (171 nm ± 4.4, PdI= 0.116) with desired cationic character (zeta
potential about +25). However, the highest encapsulation efficiency level was reached about 30% ± 0.2. AFM
images showed nanoparticles with spherical shape and smooth surface. Conclusion: Cationic PLGA
nanoparticles MTX loaded were successfully obtained by nanoprecipitation. Results implied that PLGA-PEI NP
could be a promising drug carrier MTX. The encapsulation efficiency smaller than 50%, occurred due to the
nanoprecipitation method.
Financial Support
Acknowledgments
Capes/Brazil (Barboza IR, Sousa DL, dos Santos-Silva E, Caland LB,)
246
COMPLEXATION AND SPECTRAL, MORPHOLOGICAL AND CRYSTALLOGRAPHIC
CHARACTERIZATION OF Copaífera multijuga HAYNE OIL WITH CYCLODEXTRINS
Emanuella de Aragão Tavares
Sofia Santos da Silva
Wliana Alves Vitorino da Silva
Magda Rhayanny Assunção Ferreira
Arnobio Antônio da Silva Junior
Luiz Alberto Lira Soares
Waldir Florencio da Veiga Junior
Universidade Federal do Amazonas (UFAM)
Adley Antonini Neves de Lima
Introduction: Commonly known as copaíba oil (CO), the extract of Copaifera multijuga Hayne trees is
an oilresin used due its anti-inflammatory and antinociceptive properties. Complex with cyclodextrins (CDs) is a
strategy to improve the low solubility of oils, subsequently, its oral bioavailability. This work aimed to form
inclusion complexes with B-ciclodextrin (BCD) and 2-Hydroxypropyl-B-ciclodextrin (HPBCD) and CO comparing
them by physical chemical assays. Methods: Gas Chromatograph (CG) with FID using standard B-cariophyllene
quantified this compound in C. multijuga oil. Complexes of CO:BCD and CO:HPBCD were obtained by physical
mixture (PM), kneading (KN) and slurry (SL), in 1:1 molar ratio, considering the molar weight of B-cariophyllene.
The samples were analyzed by Fourrier Transform Infrared Spectroscopy (FTIR) with an ATR and spectral
correlation by ad hoc algorithm (SC). X-Ray powder diffraction (XRD) and Scanning Electron Microscopy (SEM)
methods evaluated the crystallinity and morphology of the mixtures. Results: CG analysis indicates that the
concentration of B-caryophyllene in the volatile fraction (32.54% v/v) of copaiba oil is equal to 45.93% ± 0.2437
(0.54%). CO spectral analysis identified bands in 884 cm-1(=CH) and in 2927 and 2856cm-1 (-CH2). Differences
between CO and the complexes spectra are evident, but, the contrast between the spectra of PM, KN and SL are
less noticeable in all complexes. SC with both methods showed similar profiles with significant correlations in
3356 cm-1 (0.77) in KN, and 3298 cm-1 (0.58) in SL. In XRD evaluation the tree complexes has shown similar
amorphous profile to HPBCD and decrease in the crystalline peaks of BCD. These results corroborate with the
SEM micrographs and show differences in the characteristics of cyclodextrins and complexes formed.
Conclusion: The obtained results by FTIR, XRD and SEM analyses shown that SL has a greater number of
interactions between the BCD and CO, indicating more complexation.
247
INCREASE SOLUBILITY OF NEW ANTIFUNGAL NAPHTOQUINONE IN INCLUSION
COMPLEXES WITH CYCLODEXTRINS
Fabia Julliana Jorge de Souza
UFRN
Elen Diana Dantas
William Nascimento Litaiff Nogueira
Cláudia Cândida Silva
Universidade Estadual do Amazonas
Emerson Silva Lima
Vitor Francisco Ferreira
Universidade Federal Fluminense (UFF)
Introducion: The ciclofuronodiona (CFND) is a new quinone with antifungal potential and has shown low
aqueous solubility. This study has the intention to obtain inclusion complexes (IC) with B-cyclodextrin (BCD) and
methyl-B-cyclodextrin (MBCD) to increase solubility of this new quinone. Methods: Phase diagram was performed
to determine IC stoichiometry. It was obtained by physical mixture, kneading and evaporation (EV). Characterized
by Thermo gravimetric analysis, Differential thermal analysis and Differential Scanning Calorimetry
(TGA/DTA/DSC), Fourier transform infrared spectroscopy (FTIR) with spectral correlation by ad hoc algorithm,
Scanning electron microscopy and X-ray powder diffraction. Results and Discussion: The phase diagram
presented a linear profile. The DSC of CFND showed two initial events, one endothermic of fusion (194°C),
followed by a exothermic event of recrystallization (204°C). The decomposition happened in an endothermic
event, between 245-251°C, followed by a weight loss on TG. DSC showed a decrease in enthalpy of fusion and
recrystallization events and displace the degradation event, characteristics of CFND, changes more evident with
MBCD IC especially by EV. FTIR showed interactions in IC, changing the finger print CFND spectrum. The
spectral modifications were showed when analyzed with the ad hoc algorithm, more significative in MBCD IC.
EV-MBCD showed the best results with 74.4% of crystallinity decrease compared to CFND and the micrographs
showed the most amorphous morphology compared to other IC obtained. Conclusion: The results indicated
physical-chemical, spectroscopic, thermodynamics and morphologic modifications and decrease on crystalline
form in relation to CFND. The best results were obtained with the MBCD IC EV and showed solubility increase of
this CFND.
Financial Support
CNPQ
Acknowledgments
Pedro Henrique Antunes de Azevedo (in memoriam)
248
MUCOADHESIVE PROPERTY AND SERYNGEABILITY OF THERMORESPONSIVE
SYSTEMS CONTAING HYPERICIN
Fernanda Belincanta Borghi Pangoni
Mariana Volpato Junqueira
Marcos Luciano Bruschi
Hypericin (Hyp) is a natural photoactive pigment originated from plants of the Hypericum perforatum L.
which has been extensively studied for many photobiological activities. In vivo and in vitro studies have
demonstrated the antitumoral activity from Hyp using Photodynamic Therapy (PDT). Polymeric systems
composed by Carbopol 934P (Carb) was extensively used due to their mucoadhesive properties. In this sense,
the use of Carb associated with Poloxamer 407 (Polox) presented to be a good system to be tested in order to
collaborate to colorectal cancer treatment with PDT association. The formulations with and in absence of Hyp
were prepared with polymeric dispersion of Carb (0.15%, w/w) in distilled water with following addition of Polox
(20%, w/w), triethanolamine as the neutralizing agent and then the addition of Hyp 0.01% (w/w). Mucoadhesive
strength was evaluated using TA-XT2 plus Texture Analyser, in tension mode with mucin disc attached to the
lower end of the probe. The force required to detach the mucin disc from surface of formulations was determined
from the force-distance graph. The same equipment was used for syringeability test. In compression mode, a
syringe was then vertically clamped, and the probe lowered until initial contact with the plunger of the syringe was
observed. The resistance to expression of the syringe contents was determined by force - distance plot. The
influence of Hyp presence on the properties were statistically evaluated using ANOVA (p<0.05). The presence of
Hyp increased the mucoadhesive force (0.508 to 0.365 N). The work necessary to expel the system from a
syringe was 58.64 N.mm for the system without Hyp, and 70.81 N.mm for the Hyp system. This result showed to
be was satisfatory, since the suitable values are between 20 and 380 N.mm. Therefore, the results displayed that
Hyp formulation was appropriate to be applied by a probe in colorectal region, and exert good adhesion.
Financial Support
CAPES, CNPq, Fundação araucária, FINEP, UEM.
Acknowledgments
CAPES, CNPq, Fundação araucária, FINEP, UEM.
Ethical approval
OK
249
Characterization of mucoadhesive nanotechnological formulations containing
imiquimod for vaginal application.
Flávia Pires Peña
Luiza Abrahão Frank
Rafaela Pletsch Gazzi
Introduction:Imiquimod is a drug used in the treatment of cervical câncer, but it has adverse effects
such as irritation and pain.Nanotechnology proposes a way to deal with the obstacles caused
by
drug.Formulations administrated vaginally need to be mucoadhesive so the drug can achieve ahigher effect.So,
this work aims to develop adhesive nanoparticles (coated or not with chitosan) and incorporate this new
formulation in different hydrogels for vaginal application.Methods:Polymer formulations were prepared by
nanoprecipitation method and named as NCimiq (nanocapsules with imiquimod) and NCimiq-chit (nanocapsules
coated with chitosan).The diameter of the particles was analyzed by laser diffraction technique (MastersizerR
2000,Malvern) and electrophoretic mobility (Nano-ZS model ZEN 3600,Malvern, USA).The pH of the formulations
was obtained using Micronal B-474 pot. The hydrogels of chitosan (GELchit) containing NCimiq and hydrogels of
hydroxyethylcellulose (GELhec) containing NCimiq-chit were obtained by the manual method.Results and
Discussion:The results show that the average particle size was 337±8nm for NCimiq and 268±5nm for NCimiqchit.50% of the particles of NCimiq have 118±5nm and NCimiq-chit have 78±3nm.90% of the particles of NCimiq
have 226±3nm and NCimiq-chit have 160±2nm.It was observed that both formulations have adequate
homogeneity and unimodal behavior.The zeta potential obtained for NCimiq was -2mV±0.4 and for NCimiqchit,10,6mV±0.3.The pH obtained was 5,9±0,4 for NCimiq and 4,6±0,4 for NCimiq-chit. It was possible to verify
the presence of coating on the wall of the particles, since there was a change of zeta potential after coating
formulation NCimiq-chit.The pH of the GELchit was 4.3±0.3 and GELhec was 5.5±0.4 and is compatible for
vaginal administration.Conclusions:The polymeric nanocapsules developed have appropriate nanotechnological
features as well as the hydrogel formulations and have a potential use for the treatment of cervical cancer.
Financial Support
CAPES, CNPq
250
STABILITY AND ANTIFUNGAL ACTIVITY OF AN AMPHOTERICIN B MICROEMULSION
SYSTEM
Francine Johansson Azeredo
Faculty of Pharmacy, Federal University of Bahia, Salvador, Ba, Brazil.
Sarah Rafaelly dos Santos
Pharmacy Graduate Program, Federal University of Rio Grande do Norte, Natal, RN, Brazil.
Caroline Aoqui
Departament of Pharmacy, Dispersed Systems Laboratory (LasiD), University of Rio Grande do Norte, Natal, RN, Brazil.
Jéssica da Silva
Departament of Pharmacy, Dispersed Systems Laboratory (LasiD), University of Rio Grande do Norte, Natal, RN, Brazil.
Walicyranison da Silva
Guilherme Chaves
Eryvaldo Sócrates Tabosa do Egito
Introduction: Microemulsions systems (MEs) have a high importance mainly because they can act as
an alternative pharmacological treatment that is more efficient and with reduced adverse effects, which allows the
incorporation of drugs that are not widely used anymore due to their potential toxicity. The aim of this study was
to investigate the stability and antifungal activity of a ME containing Amphotericin B (AmB) and to compare its
performance with micellar formulations of AmB. Methods: AmB was directly incorporated in the ME containing a
water phase, an oil phase and co-surfactants under magnetic stirring. AmB-ME, blank ME, M-AmB-Sol, and MAmB-Powder were used in this study. Samples were kept in storage at two different temperature (25 and 45 ºC)
for 90 days. At predetermined time intervals, AmB content and its efficacy were evaluated. The quantification of
AmB was performed by HPLC with UV detection at 408 nm. Drug degradation models were tested by fitting the
experimental data to their appropriate equations. The efficacy of all formulations was evaluated against C.
parapsilosis ATCC 22019 using the broth microdilution method in triplicate. Results and Discussion: When
comparing different models, the best correlation (r2 = 0.97) was observed when the data were adjusted to the
first-order equation. AmB-ME showed a shelf life of approximately 26 days when stored at 25 °C and protected
from light, whereas M-AmB-Sol and M-AmB-Powder was stable for 17 and 8 days, respectively. In high storage
temperature the degradation was higher for all formulations. M-AmB-Sol formulations were able to provide fungus
inhibition until day 7 while M-AmB-Powder and AmB-ME formulations were able to provide inhibition of fungal
growth through all the experiment. Conclusions: AmB-ME has higher stability than M-AmB-Sol and M-AmBPowder formulations. Moreover, the activity against C. parapsilosis was higher for AmB-ME, providing a growth
inhibition during the 90 days of this study.
Financial Support
Cnpq and Capes
251
DEVELOPMENT AND CHARACTERIZATION OF ATORVASTATIN NANOPARTICLES
Gabriel pedroso Viçozzi
Universidade Federal Do Pampa-UNIPAMPA
Eduardo Andre Bender
Letícia Marques Colomé
INTRODUCTION: Cardiovascular diseases are getting more frequent in the global population. These
pathologies have many factors, such as stress, incorrect eating habits and smoking. From this perspective, more
methods to prevent and treat these types of pathologies need to be developed. The use of drugs statins as
hypocholesterolemic agents is new and useful. Among the different types of statins, atorvastatin is a drug that is
most effective, acting as an inhibitor of the early steps of cholesterol biosynthesis by inhibiting the enzyme HMGCoA reductase converting the HMG- CoA substrate mevalonic acid. METHODS: Among the possible
alternatives, there are the preparations of polymeric nanoparticles containing atorvastatin by precipitating
technique of preformed polymer. Thus, nanocapsules containing atorvastatin were made and the average particle
size was evaluated by laser diffraction, The pH of the particles was measured in phmetrer, the quantification of
the particle was made by HPLC in the following parameters: column C18 size 12cm, mobile phase acidified water
PH 3 with phosphoric acid + Acetonitrile + methanol (30 %, 10 % and 60 % respectively) running time of 6min
and flow 1mL/min. The parameters were, dosing of atorvastatin and encapsulation profile. RESULTS AND
DISCUSSION: The average particle diameter of the particles were D [0.5] 190nm and polydispersity (SPAN) 1.9,
these results demonstrate a satisfactory size of particles and a proper polydispersity, demonstrating how the
particle keeps stable even encapsulated and with a nanosize. The obtained calibration curve has a R²=0.9987,
demonstrating a linear analytic curve, the dosing of atorvastatin was 91% and the encapsulation profile was 93%.
The Ph of the particle was 7.1. CONCLUSIONS: In this study we conclude that the preparation of nanocapsules
containing atorvastatin has a reduced size, without destabilizing the particle structure and the dosing of
atorvastatin and encapsulation profile are adequate to.
Financial Support
Programa de Desenvolvimento Acadêmico - PDA 2015 UNIPAMPA
Acknowledgments
Programa de Desenvolvimento Acadêmico - PDA 2015 UNIPAMPA
252
CARVACROL/B-CYCLODEXTRIN INCLUSION COMPLEXES: PHYSICOCHEMICAL
CHARACTERIZATION AND IN VITRO ANTILEISHMANIAL ACTIVITY
Pharmacy Department,Federal University of Sergipe, Sergipe, Brazil.
Igor Araújo Santos Trindade
Lucindo José Quintans-Júnior
Physiology Department, Federal University of Sergipe, Sergipe, Brazil.
Daniel Pereira Bezerra
Gonçalo Moniz Research Center, Oswaldo Cruz Foundation, Salvador, Bahia, Brazil.
Danielle Lazarin-Bidóia
Graduate Program in Pharmaceutical Sciences, State University of Maringá, Paraná, Brazil.
Celso Vataru Nakamura
Graduate Program in Pharmaceutical Sciences, State University of Maringá, Paraná, Brazil.
The inclusion of carvacrol into B-cyclodextrin (B-CD), its physicochemical characterization and
antileishmanial activity was studied. Solid inclusion complexes were prepared by physical mixture (PM),
ultrasound (US) and freeze drying (FD) methods. Physicochemical characterization of complexes was carried out
using differential scanning calorimetry (DSC), thermogravimetric analysis (TG/DTG), Karl Fischer titration (KFT)
and scanning electron microscopy (SEM). The entrapment efficiency (EE%) of the inclusion complexes was
determined using high performance liquid chromatography (HPLC). Finally, the study evaluated the in vitro
antileishmania effects of free carvacrol and its inclusion complexes against the promastigote and amastigote
forms of Leishmania amazonensis. The results of DSC for US and FD methods did not show any endothermic
peak in the temperature range of carvacrol's volatilization, indicating the inclusion of carvacrol inside βCD cavity
and their TG/DTG curves showed that were able to incorporate great amount of carvacrol, as evidenced by the
mass loss of 7.27 and 8.70%, respectively (169-305°C). SEM images showed decreased size of the crystals,
especially in the US. HPLC analyses demonstrated that higher EE was obtained for FD method (81.2%). In vitro
studies of free carvacrol indicated an IC50 of 55.7 ± 3.2 µg/mL against the promastigote form and 2.9 ± 0.5 µg/mL
against the axenic amastigote form of Leishmania amazonensis. However, the calculated IC50 values to
inclusion complexes against the promastigote form was >100 µg/mL and only FD method showed significant
activity against amastigote forms. Cytotoxicity in J774A1 macrophages analysis to free carvacrol exhibited a
CC50 of 230 ± 7.3 µg/mL, which is more cytotoxic than the FD method values (CC50 >1000 µg/mL). Therefore,
the inclusion of carvacrol in βCD positively influences it's in vitro antileishmanial activity, represent a promising
alternative for the treatment of leishmaniasis.
Financial Support
CAPES, CNPq and FINEP
Acknowledgments
CNPq, CAPES, Oswaldo Cruz Foundation and Tecnologic Innovation Laboratory for Development Drugs and Cosmetics of the State University of Maringá
253
SIMVASTATIN-LOADED CHITOSAN NANOCAPSULES FOR AN INNOVATIVE
ADMINISTRATION OF STATINS
Gabriela Garrastazu Pereira
Gabriele Dadaut Souto
Fabio Sonvico
Università degli Studio di Parma
I. Background Statins when given orally undergo extensive hepatic metabolism leading to low
bioavailability II. Aim The purpose of the present study was to prepare and evaluate physico-chemical properties
of simvastatin-loaded nanoparticles suitable to oral and trans-mucosal delivery in view of an improved
bioavailability III. Methods Chitosan and lecithin and two oily excipients (medium chain triglicerides MCT and
glycerol monlinoelate GML kind gift of Trapeze) were used to prepare nanocapsules loaded with simvastatin (SIM,
1 mg/ml). Nanoparticles were characterized for particle size and surface charge (ZetaSizer NS), morphology
(STEM, EVO), drug encapsulation efficiency and drug release (HPLC). IV. Results Nanoparticles showed average
particle size between 185 and 272 nm. Chitosan/lecithin nanospheres produced without oil showed the largest
size (272 nm). All other SIM-loaded nanocapsules showed particle size below 200 nm, independently from oil type
and concentration. Furthermore, formulations prepared had positive surface charge and narrow size distribution.
SIM encapsulation efficiency was low 22% in absence of the oil components. Adding MCT as oily phase
increased the encapsulation up to 74%; while using GML, the encapsulation efficiency attained 86%. However, it
was the combination of both excipients that achieved the highest encapsulation efficiency 94%. This latter
formulation showed a prolonged release in the conditions selected (40% SIM released after 48 h in PBS at 37oC)
V. Summary Chitosan nanocapsules for simvastatin drug delivery have been developed and will be studied for
improving statins bioavailability by oral or alternative transmucosal routes
Financial Support
Cnpq
254
SIMVASTATIN-LOADED POLYMERIC NANOPARTICLES FOR PULMONARY DELIVERY
SYSTEM
Gabriele Dadalt Souto
Fabio Sonvico
Universita' degli Studi di Parma
Introduction: Recent research indicates that statins have additional anti-inflammatory and
immunomodulatory properties, which are being studied for their possible role in the treatment of various respiratory
disorders. Polymeric nanocapsules (NC) are delivery systems composed of an oily core surrounded by a thin
layer of biodegradable polymer, which allows controlled drug release, drug stabilization and increased
bioavailability. In this way, the purpose of the present study was to prepare and evaluate pulmonary drug delivery
systems composed of simvastatin-loaded polymeric nanocapsules (NCSV). Methods: NCSV were prepared by
interfacial deposition of preformed polymer, containing poly(εcaprolactone), sorbitan monostearate, capric/caprylic
triglycerides, simvastatin and polysorbate 80. Results and Discussion: NCSV exhibited average particle size of
205.5±0.5 nm, with slightly acid pH (6.06±0.08) and negative zeta potential (-19.5±0.13 mV). The analysis by
fourier transformed infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC) indicated that no
chemical reactions occurred among components during the development of the formulations. The DSC analysis
showed that sorbitan monostearate was dissolved in the oil core of nanoparticles. The X-ray diffractometry,
scanning electron microscopy (SEM), and optical microscopy indicated that SV is molecularly dispersed in the
NC. The loading efficiency of SV was 99.18±0.72%. The aerosolization performance was determined using Next
Generation Impactor (NGI) at a flow rate of 15 L/min. Different nebulizers (Pari e-flow and Pari Turbo-boy) were
employed to deliver and to aerosolize NCSV and SV suspension. The mass median aerodynamic diameter
(MMAD) and the fine particle fraction (FPF < 5 µm) were determined according to the USP 34. Conclusion:
NCSV were successfully delivered with a FPF = 61.9% using a jet-neb nebulizer while the nano-suspension could
not be aerosolized using a mesh technology nebulizer.
Financial Support
CNPq, CAPES
255
OVERCOMING UNPLEASANT TASTE OF METFORMIN HYDROCHLORIDE BY COLD
EXTRUSION/SPHERONIZATION EMPLOYING SOLID LIPID BINDERS
Gustavo Freire Petrovick
Institute of Pharmaceutics and Biopharmaceutics, Heinrich-Heine-University, Düsseldorf, Germany
Miriam Pein
Jörg Breitkreutz
Metformin is used in treatment of type II diabetes mellitus. Tablet sizes and bitter taste of this API,
especially when tablets are crushed, have a negative impact on patient compliance. Solid lipids as excipients for
oral dosage forms have advantages of lack of toxicity and good taste-masking attributes. In the present study, a
recently improved extrusion/spheronization method for solid lipids should be applied to produce taste-masked
pellets containing metformin HCl. Extrusion of solid lipid mixtures containing 50-80% of metformin were performed
using a co-rotating twin-screw extruder. Batches of extrudates were rounded using a spheronizer. An IR light was
used to heat the material until 33 °C. Taste masking assessment was performed using a taste sensing system.
API solutions were analyzed prior to measurement for equipment calibration. Pellets were stirred in water for 30
and 60 s, were filtered and measured. A linearity was observed for all sensors responses, starting at
concentrations of 0.50 mg/mL, indicating good sensor sensitivity towards API. A principal component analysis
(PCA) was performed using the sensor responses to the pellets. The PCA map showed that pellets based on
higher amounts of lipids presented more similar responses to the drug-free formulation than to pure API, indicating
more look-like taste properties. Samples that released API over 60 s were located closely to the API than those
released over 30 s. To simplify the differentiation of the formulations, Euclidean distances were calculated. The
distance between API and placebo was considered as 100% taste variation. Pellets based on hard fat only
present better taste-masked properties compared to pellets with ternary lipid mixtures. Results of an electronic
tongue evaluation proved that the investigated extrusion/spheronization technique was suitable to overcome the
unpleasant taste of metformin HCl producing taste-masked lipid pellets containing high drug load of the API.
Financial Support
DAAD ¿ German Academic Exchange Service
Acknowledgments
none
Ethical approval
was is not necessary, the experiments were in vitro
256
DEVELOPMENT OF LIPID NANOCARRIER NASAL DRUG DELIVERY SYSTEM
CONTAINING CURCUMIN FOR BRAIN TARGETING
GUSTAVO RICHTER VAZ
Universidade Federal do Rio Grande (FURG)
GABRIELA HÄDRICH
JULIANA BIDONE
HELDER FERREIRA TEIXEIRA
JEAN-LUC PUTAUX
Université Grenoble Alpes (UGA)
JOSÉ MARÍA MONSERRAT
ANA LUIZA MUCCILLO-BAISCH
ANA PAULA HORN
ANTÔNIO SÉRGIO VARELA JUNIOR
CRISTIANA LIMA DORA
Introduction: Antioxidant compounds, such as curcumin (CUR), have interesting properties that can be
useful in the treatment of neurodegenerative diseases. However, studies revealed that CUR presents low
bioavailability and the development of curcumin-loaded nanocarrier systems may be considered as a promising
way to exploit its therapeutic properties. The aim of this study was to develop lipid nanocarriers nasal drug
delivery systems containing curcumin for brain targeting. Methods: The nanocarriers (NE) were prepared by the
hot solvent diffusion method associated with the phase inversion temperature. Physico-chemical and
morphological characterizations were performed and in vitro antioxidant activity was evaluated by DDPH method.
The permeation of CUR was analyzed in Franz type diffusion cell using porcine nasal mucosa and confocal laser
scan and histologiccal studies were also performed. Results and Discussion: The results indicated that the NEs
size were ranging between 18 nm and 44 nm and the zeta potential was negative. The TEM and Cryo-TEM
analysis indicated that the formulations were homogeneous and had the sizes in accordance to the dynamic light
scattering analysis. The CUR content ranged from 0.24 to 1.50 mg/mL, and the encapsulation efficiency was
greater than 99%. The DPPH assay demonstrated that the antioxidant capacity of CUR remained unchanged
after the nanoencapsulation process. CUR-NE permeation across the porcine nasal mucosa was higher than free
curcumin. The analyzes carried out on confocal microscopy allowed to prove that the permeation CUR-NE was
better than the free CUR and no toxicity for nasal mucosa was observed in histological analysis. Conclusion:
These results suggest that it was possible to develop nanocarriers with high content of curcumin and small
particle size and that the encapsulation improved its potential of permeation across the porcine nasal mucosa.
Financial Support
Edital MCTI/CNPq/MEC/Capes ¿ Casadinho/ Procad 2011. Processo CNPQ: 552457/2011-6
257
Development and characterization of surface modified PLGA
nanoparticles for potential application in cancer therapy
Renata de Carvalho Feitosa
Lília Basílio de Caland
Introduction: Colloidal nanocarriers improves the efficacy and reduces the side effects of drugs and
biomolecules applied for cancer therapy. Some functionalized cationic biodegradable nanoparticles (NP) offer the
possibility to overcome biological barriers and target specific tissues. Parameters such as size, shape and
surface characteristics of particles seems to play a critical role in the binding to cell membrane and cellular
uptake, especially with respect to tumors treatment. Among the drugs used for this purpose, doxorubicin (DOX) is
a potent antineoplastic used against a wide spectrum of solid tumors. Method: In this study, NP were prepared
and optimized by nanoprecipitation. Different surfactants (polysorbate 80, 85, sorbitan monooleate 80, polyvinyl
alcohol (PVA), Poloxamer 188 and Poloxamer 407), as well as distinct poly (lactic-co- glycolic acid) (PLGA)/ratios
were tested to obtain small and narrowed sized nanoparticles. The best formulation was cationic functionalized by
using polyethyleneimine (PEI 25 kDa) in a PEI:PLGA ratio of 1:5. After assure the biocompatibility, fluorescent
NP were prepared with PLGA labeled with fluorescein isothiocyanate (FITC). Results and Discussion: The best
exhibited particle size of 59.73 nm ± 7.63 (PdI= 0.148 ± 0.003) and zeta potential of +17.09 mV ± 3.66, which was
biocompatible in VERO cells. This formulation was selected for the encapsulation efficiency experiments.
Furthermore, the fluorescence of the particles were demonstrated evidenced by fluorescence microscopy image,
Fourier Transforms Infrared (FTIR)-ATR, and by using flow cytometry. Conclusions: The fluorescent and cationic
biodegradable nanoparticles (PLGA-PEI-FITC NP) were successfully optimized for antitumor drug encapsulation.
Financial Support
Acknowledgments
Capes/Brazil (Barboza IR, Sousa DL, dos Santos-Silva E, Caland LB,)
258
DEVELOPMENT, CHARACTERIZATION AND EVALUATION OF BIOADHESIVE
DELIVERY SYSTEMS (BDS) WITH OILRESIN (OR) IN THE TREATMENT OF
PERIODONTAL DISEASES
Irina dos Santos Miranda Costa
Mônica Freiman de Souza Ramos
Osvaldo de Freitas
Maria Paula Garofo Peixoto
Carolina Patrícia Aires
Introduction: Periodontal diseases have high prevalence and incidence, being considered a public health
problem. They are an infectious process which if not treated can lead to tooth loss, affecting the gums and bone
that support the teeth. Treatment consists of removal of the biofilm by mechanical debridement through scaling
and root planning and bacterial elimination using antimicrobial agents. Metronidazole (MTZ) is a 5-nitroimidazole
with potent antimicrobial activity. The systemic use of antibiotics causes various adverse effects and bacterial
resistance. The copaiba oil (OC) is an OR widely used in folk medicine, and studies have shown its promising
effect on the inhibition of the main bacteria in dental plaque, besides having film property. Thus, a local BDS
would be an effective alternative, providing greater retention, allowing the drug dosage control, and promoting
prolonged release. The objective of this work was the development of a bioadhesive polymeric film with MTZ and
OR as an alternative to conventional therapy. Methods: Films were prepared using the solvent casting technique.
The mixture consisting in dispersion of gelatin and chitosan; 25% MTZ; and OR (1; 2.5; and 5% OC; 10% of
diterpene fraction (FD)). Drug crystallization was observed under a microscope and the mechanical properties
were measured using a texture analyzer. Thickness was obtained using a digital micrometer and the mass was
determined. Results and Discussion: The microscopy showed that there was incorporation of MTZ, OC and FD in
the polymer dispersions, however, at higher concentrations of OC crystallization of MTZ was observed in the film
surface. The films with 1% OC and FD were the most promising regarding morphology, demonstrating the
influence of OR on the mechanical properties of the films as well as the thickness measurements and mass.
Conclusions: The polymer films containing 1% OC and FD have shown promise as BDS for the treatment of
periodontal diseases.
Financial Support
Capes
259
INCLUSION COMPLEXES OF beta-CYCLODEXTRIN AND Lippia pedunculosa
ESSENTIAL OIL
ISLA ALCÂNTARA GOMES
FRANCIELLY DE OLIVEIRA ARAÚJO
BRUNA MARIA HIPÓLITO DE SOUSA
IGOR ARAÚJO SANTOS TRINDADE
YASMIM MARIA BARBOSA GOMES DE CARVALHO
EMMANOEL VILAÇA COSTA
Departament of Chemistry, Federal University of Sergipe, São Cristóvão, SE, Brazil.
SÍLVIA STANISÇUASKI GUTERRES
Faculty of Pharmaceutical Sciences of Federal University of Rio Grande do Sul, Porto Alegre, RS, Brazil.
MAIRIM RUSSO SERAFINI
Introduction: The essential oil extracted from Lippia pedunculosa (EOLP) consists of secondary
metabolites of pharmacological interest, however essential oils are photosensitive and volatilize quickly.
Accordingly, cyclodextrins (CDs) are technological alternatives to improve stability and solubility. Methods: The
complexes were obtained by physical mixture (PM), paste complex (PC) and slurry complex (SC). The
identification of the chemical constituents of EOLP was performed by gas chromatography coupled to mass
spectrometry (GC/MS) and the samples were characterized using differential scanning calorimetry (DSC),
thermogravimetry/derivative thermogravimetry (TG/DTG), scanning electron microscopy (SEM), Fourier transform
infrared spectroscopy (FTIR) analysis and particle size. Results and Discussion: The analysis obtained by
GC/MS revealed that the rotundifolone (72.02%) was the major constituent of EOLP. The oxidative DSC curve
demonstrated the oxidation of the oil, in detriment of the other samples, suggesting that beta-CD protects it from
oxidation. The TG/DTG suggests that the best method of inclusion was the SC, because of its greater weight
loss (11.6 %) in the second stage (171-312 °C), while the PM had the lowest mass loss (1.9 %) in the same
range, suggesting low complexation. The SEM showed changes of the typical crystalline form of PC and SC
methods. The particle size distribution obtained by laser diffraction showed that the PC (2.87±0.51) and SC
(3.00±1.87) particle size was smaller when compared to PM (1.37±0.06). Moreover, the infrared spectra showed
the presence of the rotundifolone functional groups in PC and SC. The TG/DTG curves showed increase weight
loss in the second stage, relative the increase evaporation temperature, indicating replacement of the water
molecules present in the internal cavity of the beta-CD by EOLP. Conclusions: Based on these results, was
suggested complexation between EOLP and beta-CD by SC.
Financial Support
The authors are grateful to CAPES, CNPq, FINEP and FAPITEC/SE for financial support and fellowships.
Acknowledgments
We thank Maria Lúcia Vieira Moreno, Cláudio Pereira Figueira and Adriana Lanfredi Rangel for the images of SEM from the Gonçalo Moniz Research Center,
Oswaldo Cruz Foundation, Bahia, Brazil.
260
POLYMERIC NANOCAPSULES CONTAINING PEQUI OIL (Caryocar brasiliense
Cambess)
Jader Büttner-Pires
Pharmacy Graduate Program, University of Mato Grosso, Sinop, MT, Brazil
Faculty of Pharmacy, University of Rio Grande do Sul, Porto Alegre, RS, Brazil
Faculty of Pharmacy, University of Rio Grande do Sul, Porto Alegre, RS, Brazil
Adilson Paulo Sinhorin
ICNHS, University of Mato Grosso, Sinop, MT, Brazil
Stela Regina Ferrarini
Pharmacy Graduate Program, University of Mato Grosso, Sinop, MT, Brazil
Introduction The pequi oil has proven antioxidant activity. Polymeric nanoparticles have been
extensively studied as potential drug carriers, reflecting the need for drug release in specific body areas, reducing
the therapeutic dose of the drugs, decreasing side effects and improving the therapeutic ones. Methods The
lipophilic nanoparticles were prepared by interfacial deposition of poly(εcaprolactone), caprylic/capric triglyceride
and sorbitan monoestearate as the lipid core and pequi oil (1.0 mg.mL-1), after coated with chitosan. Formulations
were characterized according to their size distribution (laser diffraction, dynamic light scattering, zeta potential
(electrophoretic mobility) and TEM. Thereafter, semissolid formulations were produced (gel) containing pequi oil
(0.10%) and/or the nanostructured ingredient (5%). These formulations were characterized by their organoleptic
characteristics, pH, rheological behavior. Results and Discussion The values of D[4.3], obtained by laser
diffraction, ranged between 138 nm and 141 nm. The mean size, obtained by DNS and zeta potential ranged
between 141 nm and 159 nm and 19,6 mV and 25,3 mV, respectively. The lipophilic nanoparticles, as a milky
white suspension, showed a Newtonian rheological behavior. All gel formulations presented pseudoplastic
rheological behavior. The addition of nanoparticles did not alter their rheological behavior. The semi-solid
formulations presented pH adequate for cutaneous application, being around pH 4,12, regardless the presence of
the nanostructures and pequi oil. Conclusions The nanostructured natural ingredient presented particle size at the
nanoscale, stable inputs being suitable of use in formulations like semissolids. The semissolid formulations
containing or not the actives showed suitable physic-chemical characteristics for cutaneous application. The
rheological behavior was not affected by the presence of the nanoparticles in semissolid formulations.
Financial Support
FAPEMAT-Fundação de Amparo à Pesquisa do Estado de Mato Grosso
Acknowledgments
FAPEMAT-Fundação de Amparo à Pesquisa do Estado de Mato Grosso, UFMT-University of Mato Grosso and UFRGS-University of Rio Grande do Sul
261
RHEOLOGICAL INTERACTION PARAMETER OF BIOADHESIVE THERMORESPONSIVE
SYSTEMS COMPOSED BY POLOXAMER 407 AND CARBOPOL 974P
Jéssica Bassi da Silva
State University of Maringa
Sabrina Barbosa de Souza Ferreira
Polymeric blends containing bioadhesive and thermoresponsive polymers are considered as potential
drug delivery systems. The rheological synergism has been reported as a form to evidence the adhesive
interactions between the polymers. Therefore, the aim of this study was to evaluate the rheological interaction
parameter of formulations composed by poloxamer 407 (P407) and Carbopol 974P (C974P) in order to select the
best systems to be used in the development of drug delivery systems. Monopolymeric systems and binary
polymeric systems containing P407 (15 or 20%, w/w) and C974P (0.10, 0.15, 0.20, 0.25 or 0.50%, w/w) were
prepared, totalizing seventeen formulations. Oscillatory rheometry was performed using a MARS II (Haake®)
controlled stress rheometer in oscillatory mode at 5, 25 and 37 ± 0.1 °C. After the determination of the linear
viscoelastic region, frequency sweep analysis was evaluated from 0.1 to 10.0 Hz. The viscoelastic properties of
at least five replicates were calculated using RheoWin 4.10.0000 (Haake®) software. A possible rheological
interaction parameter between the polymers was evaluated by the difference between the dynamic modulus of the
polymeric blends and the sum of the individual parts at the frequency of 10.0 Hz. Only the rising C974P
concentration leads to a significant increase the rheological interaction parameter, which not observed to the P407
concentration. Thus, formulations with 0.25 and 0.50% (w/w) C974P resulted in higher rheological value for blends
with 15% (w/w) P407. The binary polymeric systems 15/0.20, 15/0.25, 15/0.50 and 20/0.10, at 37 °C, showed
positive interaction parameter. Higher temperature values were associated with higher interaction parameter. The
results evidenced the 15/0.25 formulation was the most suitable to the development of pharmaceutical systems
and was choosing to validate mucoadhesive methodologies, since presented the higher rheological parameter
interaction value at 37 °C.
Financial Support
CAPES, CNPq, FINEP and UEM
262
MICROSPHERES CONTAINING ANTIVIRAL DRUGS: DEVELOPMENT,
CHARACTERIZATION AND IN VITRO DRUG RELEASE PROFILES EVALUATION
Jéssica Brandão Reolon
Universidade Federal do Pampa (UNIPAMPA)
Maicon Brustolin
Thainá Accarini
Sandra Elisa Haas
Eduardo André Bender
Letícia Marques Colomé
The herpes simplex virus (HSV) is a major public health problem in several countries causing ulceration.
Acyclovir (ACV) is a drug that belongs to the therapeutic arsenal for the treatment of HSV infection. However
there are limitations in its use due to its short half-life and low water solubility. Curcumin (CUR) is a compound
which has demonstrated various therapeutic activities such as antiviral activity, but their oral use is also
compromised by the low solubility. In this context, the microspheres (MP) are a pharmacotechnical approach
which modifies the pharmacokinetics of the compound and offers the possibility of overcoming the difficulties of
the therapeutic use. Methods: MP formulations containing ACV and CUR were prepared by the spray drying
technique, and was composed of Eudragit®RS and HPMC as polymeric materials, and mannitol as drying
material. The obtained MP were characterized in terms of yield and of the technological flow properties of
powders, particle size by laser diffraction, morphological analysis in atomic force microscopy, and ACV and CUR
content by HPLC. The evaluation of the in vitro release profile was conducted by the method of the closed vials
using as a means of releasing phosphate buffer pH 6.8 (1% Tween 80). Results: All MP formulations presented
as an apparently fine and yellow powder. The yield was 44%, and a reduced post technological flow. MPs show
unimodal profile in size distribution evaluation, with an average size 13µm, corroborating morphological analysis
which showed the presence of particles roughened. The drug content evaluation shows mean values of 99% to
92%, to ACV and CUR, respectively. The in vitro release profile observed that the MP was able to control the
release to ACV and improve the solubility of CUR. Conclusions: This study demonstrated that the spray drying
technique was effective for obtaining MP containing ACV and CUR. The MP proved promising system for oral
administration of the compounds.
Financial Support
CAPES / Brazil, PBDA/UNIPAMPA
263
INVESTIGATION OF ELECTROSPINNING PARAMETERS FOR NON-WOVEN
PVA/CALCIUM ALGINATE CONTAINING SILVER SULFADIAZINE
Jessyca Aparecida Paes Dutra
Suzana Gonçalves Carvalho
Sarah Oliveira Lamas Souza
Postgraduate student, Department of Metallurgical and Materials Engineering, Federal University of Minas Gerais, Belo Horizonte, MG,
Brazil
Rodrigo Lambert Oréfice
Professor, Department of Metallurgical and Materials Engineering, Federal University of Minas Gerais, Belo Horizonte, MG, Brazil
Janaina Cecília Oliveira Villanova
Introduction: wound dressings are covers that provide suitable conditions for rapid healing and they act
as drug delivery system. The purpose of these work was developed a wound dressings containing silver
sulfadiazine (SSD) based on blends of calcium alginate (CaAlg) and poly(vinyl alcohol) (PVA) in order to enhance
both antibacterial activity and mechanical strength. The material were processed by electrospinning technique.
Methods: PVA and CaAlg solutions were prepared in different concentrations: PVA at 5, 10 and 12%w/v and
CaAlg at 4 and 8%w/v. The blends were obtained in different proportions of PVA and CaAlg (90/10 and
80/20%w/w). The concentrations of SSD were 0.25 and 1.0 %w/w in the blends. The mixtures were electrospun
and the diameter and the morphology of the nanofibers were characterized by scanning electron microscopy
(SEM) in order to define the ideal conditions for the process. Results and discussion: to PVA and CaAlg
concentration at 5%v/v and 4%w/v the fibers presented many beads and large diameter. CaAlg increase
concentration to 8%w/v originated fibers with smaller diameter but without beads. PVA increase concentration to
10 and 12%w/v presented no beads, oriented fibers and fiber with small dimeters. The diameter of the fibers
decreases as the concentration of the polymers in the blends increases, which suggests that the system
viscosity interfere with the formation and morphology of fibers. Furthermore, less viscous solutions originate larger
number of beads in the fibers. SSD concentration also interferes with the formation of beads and the number is
greater in higher drug concentrations. Conclusions: according the results was possible to define the
electrospinning conditions: flow rate of 1.7 mL/hour; spindle with 0.7 mm diameter; distance between the needle
tip and the ground electrode of 15 cm; and, positive/negative voltage applied of 26 kV/0 kV.
Financial Support
FAPES
Acknowledgments
The authors would like to thank FAPES, FAPEMIG and INIT/UFES.
264
ANTIMICROBIAL POTENTIAL INVESTIGATION OF NEW FLAVONE DELIVERY
SYSTEMS
Joandra Maísa Da Silva Leite
Túlio Chaves Mendes
Bruna Pereira Da Silva
Airlla Laana de Medeiros Cavalcante
Camila De Oliveira Melo
Renaly Ivyna De Araújo Rêgo
Introduction New drug delivery systems have characteristics that stand out when compared to
conventional systems, such as versatility and control the release of drugs. Because of this, many drugs have
been incorporated into these systems to improve this action against microorganisms in order to combat the
resistance of bacterium strains. Flavone is a flavonoid derivative that presents an intrinsic antimicrobial activity.
Thus, the aim of this study was to incorporate the flavone into nanosystems and analyze its minimum inhibitory
concentration against several strains. Methods Formulations with lower amounts of surfactant were selected from
the pseudoternary phase diagram developed. This study selected three systems (O/A nanoemulsion, bicontinuous
nanoemulsion and nanoemulgel), in which the flavone was incorporated (1mg/ml). For antimicrobial analysis it
was used the disc diffusion method on agar Müeller-Hinton culture medium. Four strains were used:
Pseudomonas aureginosa (ATCC 27853), Staphylococcus aureus (ATCC 25923), Enterococcus faecalis (ATCC
29212), and Staphylococcus epidermidis (ATCC 12228). The discs were impregnated with 20 µL of formulations
with and without the drug, previously placed in Petri dishes seeded and incubated at 37°C for 24h. Results and
Discussion The formulations were not effective against Staphylococcus aureus (ATCC 25923), Enterococcus
faecalis (ATCC 29212) and Staphylococcus epidermidis (ATCC 12228), but showed bacteriostatic activity against
the Pseudomonas aureginosa strain (ATCC 27853), demonstrating inhibition zones ≥ 8 mm. The bacteriostatic
activity can be observed from the no growth of bacteria on the plate. Conclusion Despite the short time analysis
to a lipid system which retains the drug for longer time, these formulations have proved to be promising
antimicrobial activity.
Financial Support
CETENE
Acknowledgments
CAPES, CNPQ
265
DEVELOPMENT AND CARACTERIZATION OF A TRICLOSAN NANOCAPSULES
SUSPENSION AIMING CERVICAL CANCER CELLS
João Guilherme Barreto De Marchi
Denise Soledade Jornada
Introduction: Triclosan is an antimicrobial and antifungal agent, which has been commonly used in
personal care products. It has been recently reported as a potential anti-cancer drug, due to its capacity of inhibits
FAS, which is relatively low expressed by normal cells, but over-expressed in cancer cells. Nevertheless its low
bioavailability and toxicity are limitations to its application. In order to improve its safety, pharmacotechnical and
pharmacological properties, we propose the use of nanotechnology to overcome all the drawbacks aiming
antitumoral formulation. Methods: Nanocapsules were obtained by nanoprecipitation followed by chitosan coating.
Physico-chemical characterizations were performed taking into account size, zeta potential, morphology, pH,
encapsulation efficiency, stability and release profile. Cell growth inhibition was determined cultivating a
standardized cervical cancer cell line (SiHa) in the presence of nanoencapsulated triclosan (NCCQ) or free drug
(TF) for 24 hours. The viable cells were detected by MTT method. All tests were performed in triplicate. Results:
Characterization results showed that nanocapsules suspension had particle diameter (131.2±1.0 nm), absence of
micrometer contamination, cationic zeta potential (13.65±0.49 mV), slightly acid (pH = 4.13±0.09), with a high
encapsulation efficiency (≈ 100%), stable within 30 days (no statistical difference between T=0 and T=30) and a
small retention of triclosan in 24 hours, when compared NCCQ (60% released) and TF (80% released).
Preliminary data showed a reduction in tumoral cells viability (≈10x) in all tested concentration (1.08 - 5.4 μg/mL),
when compared to TF or reported on literature. Conclusion: The system obtained has showed an adequate pH,
releasing profile, and antimicrobial effect for an intermediary formulation. It was able act against cervical cancer
cell lines, even when tested in lower concentration than reported on literature.
Financial Support
CNPq, Capes and FAPERGS
Acknowledgments
UFRGS and PPGCF
266
CHARACTERIZATION, in vitro ANTIOXIDANT POTENCIAL AND TOXICITY OF LAYERED
DOUBLE HYDROXIDE AND OLANZAPINE BINARY SYSTEMS
jose lamartine soares sobrinho
João Pontes Neto
Magaly Andrezza Marques Lyra
MONICA FELTS DE LA ROCA SOARES
Pedro José Rolim-Neto
The pharmaceutical industry yet faces the low bioavailability challenge of poorly water-soluble drugs in
the last decades. In this matter, Layered Double Hydroxides (LDH) present themselves as an alternative to
increasing the drug solubility rate by stabilizing its amorphous form. Olanzapine (OLZ), classified as class II in the
Biopharmaceutical Classification System, has low aqueous solubility, hindering its absorption after oral
administration. It is used in the treatment of bipolar disorder and schizophrenia, diseases that present oxidative
stress in their etiology and pathophysiology. The aim of this work was the characterization by x-Ray diffraction
and thermal analysis, in vitro evaluation of the antioxidant potential by 2,2-Diphenyl-1-picrylhydrazyl free radical
(DPPH) and thiobarbituric acid reactive species (TBARS), and toxicity evaluation by Artemia salina test. The
LDH:OLZ binary systems, obtained by the solvent technic with different drug concentrations (05, 20 and 30%),
were tested, as well as their isolated components. Comparing the binary systems, its isolated components, as
well as the physic mixture, the diffractograms patterns showed the absence of peaks related to isolated drug, in
the lower concentration (LDH:OLZ 05%), agreeing with the thermal analysis, which showed the absence of the
melting peak of OLZ, indicating the stabilization of its amorphous form. The in vitro antioxidants experiments with
the binary systems and isolated components demonstrated antioxidant capability for all samples, but the
association between LDH and OLZ promoted improvement in the drug's performance. In the Artemia salina test,
the LDH alone showed no toxicity and the binary systems decreased the toxicity of OLZ compared to the isolated
drug. These results provide good expectations for future in vivo experiments and favor the proposed use of LDHs
as a functional excipient in the pharmaceutical industry.
Financial Support
FACEPE, CNPq and CAPES
267
CHARACTERIZATION OF CLAYS FROM SAND EXTRACTION WASTE AND
INCORPORATION IN GEL AND EMULSION
Juliana da Silva Favero
Venina dos Santos
Valeria Weiss-Angeli
Rosmary Nichele Brandalise
Carlos Péres Bergmann
Introduction. The clays used in most of cosmetic products come from the clay mining process in which
occurs the dismantling of deposits and after the clay is transported to beneficiation plant. This process causes a
large environmental impact as changes in topography, erosion, deforestation, changes in water pH, etc. To
minimize these impacts, different types of clay from sand extraction waste were characterized and subsequently
associated with semi-solids vehicles. Methods. Clays (I, II, III and IV) were characterized by X-ray fluorescence
(XRF) and X-ray diffraction (XRD). The clays were incorporated into a hydrophilic gel and a non-ionic emulsion
and stored for 90 days at 20 °C ± 2 °C, 2 °C ± 2 °C and 45 °C ± 2 °C with the respective patterns (emulsion and
gel without clay). The viscosity of these systems were analyzed in the beginning and in the end of the 90 days.
Results and discussion. The XRF analysis showed silicon in the major amount followed by aluminum for all types
of clay. The XRD characterization identified kaolinite typical reflections in all samples. Clays II, III and IV
indicated illite. Clay III also presented smectite. All formulations, including the patterns, showed non-Newtonian
behavior by the linear relationship absence between stress and velocity shear values, besides thixotropy. It was
possible to observe viscosity changes in both systems, with addition of clays. Clays II, III and IV increased
emulsion viscosity while clay I decreased this parameter. The gel viscosity decreased with addition of clays II, III
and IV and increased with clay I. After 90 days of storage, the emulsions viscosity increasead, and to the gels
systems the viscosity increased or decreased according to tested conditions. Conclusion. The clays have
potential to be used in the cosmetic or pharmaceutical field by its mineralogical composition. The presence of
clays in the tested systems showed changes in viscosity.
Financial Support
Research Support Foundation of the Rio Grande do Sul State (FAPERGS)
Acknowledgments
University of Caxias do Sul (UCS) and Research Support Foundation of the Rio Grande do Sul State (FAPERGS)
268
STUDY OF NANOSTRUCTURE OF LIQUID CRYSTALLINE WITH POTENTIAL
APPLICATIONS FOR DRUG DELIVERY SYSTEM
Universidade de Sorocaba (UNISO)
Márcia Araujo Rebelo
Nathália Cristina Gonçalves de Rosa
Cecília Torqueti Barros
Francielly Cristina Côrrea Lopes
Introduction:Liquid Crystal (LC) is a physical nanostructure that behaves simultaneously as crystalline
solid and liquid. Different parameters must be considered during the obtaining LC process. This proposes work to
optimize and evaluate different processes of nanostructure systems in LC form preparation (NSLC). Methods:This
work NSLC was composed of Myverol 18-92(MO), water (18.2 MΩ.cm1) and surfactants (Tween 20 and Span
60). Different obtaining LC processes were evaluated to achieve a stable formulation. The processes differ in the
manner in which water was added, mechanical agitation and temperature. Two formulations were selected: FT
consisting of MO:tween:water (0.3:0.15:20), obtained at 45°C, the water drip process and agitation by ultrasound
and FTS with MO:tween/span:water (0.15:0.3:20) obtained the water added by in bolus process and agitation by
ultrasound at 50°C. Both formulations were stored (24h) at 25°C and analyzed polarized light microscopy. The pH,
zeta potential (ZP), particle diameter (PD) and polydispersity (PDI) were monitored (35 days). Results and
Discuss:The results correspond to the Mean±SD (n=6). The formulations FT and FTS the pH was respectively
5.74±0.43 and 6.45±0.38. For FT ZP and PD was respectively -25.04±2.18mV, 277.38±39.89nm and PDI was
between 0.261 and 0.332. For FST, ZP and PD was respectively -19.16±2.73mV, 331.35±6.78nm and PDI
between 0.302 and 0.325. For both formulations the absence of significant changes to the pH, ZP and DP is
indicative of system stability. For both formulations was observed anisotropic crystalline nanostructures that is
shaped to resemble the cubic liquid crystals. Conclusion:The structure of LC did not change depending on the
formulation composition. The pH and ZP was considered optimal for both formulations and remained stable over
35 days. For the dimensions of the LC, the FT diameter is smaller than FTS diameter. The PDI, for both, is
considered suitable for the LC structured.
Financial Support
PROSUP/CAPES, FAPESP, FINEP
269
INFLUENCE OF TEMPERATURE AND DRYING PROCESS IN THE CROSSLINKING
BIOCOMPATIBLE POLYMERS BY BUCAL ADMINISTRATION
Cecília Torquetti de Barros
Nathalia Cristina de Rosa
Franciely Cristina Correia Neves Lopes
Laboratory of Biomaterials and Nanotechnology - University of Sorocaba
Introduction: The temperature and drying processes of hydrogel films are critical factors in the
manufacture process and influence in the mechanical properties and formulations biopharmaceuticals
properties.The present study deals of influence of temperature and drying processes about the crosslinking of
blends of polymers and mechanical properties of bilayer hydrogel films (BHF).These films were designed for
administration and modified release drug. The apical layer is for a fast released and the basal layer is provided
with mucoadhesive properties and controlled drug release. Method: The polymers used for the preparation of BHF
were K4M hydroxypropyl methylcellulose (HPMC), carboxymethylcellulose (CMC) and chitosan (CH).The apical
layer was prepared with HPMC (3%) and the basal layer was prepared by mixing HPMC/CMC/CH (6:3:1
m/m).The film was obtained by extension of the hydrogels on a glass plate with extender aid.The drying
temperature of the films was adjusted to 20, 25 or 30°C.The drying conditions were environment temperature (TA),
vacuum environment (AV) and not saturated vapor pressure environment (PVNS).The influence of the drying
conditions were evaluated relative to the residual moisture content, swelling capacity, tensile strength (elongation
test), compression (mucoadhesion, relaxation and resilience) and differential scanning calorimetry (DSC). Result:
The drying time of films ranged between 2 and 312 hours. At the end of the drying process, the moisture content
ranged from 6.79% (TA 25°C) and 11.22% (AV 20°C).The swellability of BHF ranged between 0.08 ml (TA 30°C)
and 0.15 ml (TA 20°C).The mechanical properties of tensile and compression BHF were influenced by drying
conditions. The DSC results indicate absence of composite formation. Conclusion: The extension technique for
hydrogels afforded bilayer hydrogel films. The drying conditions of the formulations influence the physical,
physico-chemical and mechanical of BHF.
Financial Support
University of Sorocaba, Fapesp, Finep
270
Functionalized nanoparticles for potential benznidazole targeting for infected cells
Lannya Clara Santos Tavares Pessoa
Alaine Maria dos Santos Silva
Lília Basílio de Calland
Introduction: Benznidazole (BNZ) is the only one drug currently available for the treatment of patients
with Chagas' disease in Brazil. However, its side effects and relative low efficacy in the chronic phase is
explained mainly due to the difficulty to overcome biological barriers and low aqueous solubility. The purpose of
this study was the preparation and optimization of cationic polymethylmethacrylate (PMMA) nanoparticles for
administration route parenteral decorated with sialic acid or cholesterol for potential BNZ targeting to infected cells
by Trypanosoma cruzi parasite. The PMMA used was the Eudragit E PO. Methods: Nanoparticles were prepared
by emulsification-solvent evaporation method. The particle size, zeta potential, polydispersity index and atomic
force microscopy (AFM) images were used as performance parameters for selecting the best formulation. Drugcopolymer interactions with sialic acid and cholesterol were assessed by the encapsulation efficiency (EE%) and
Fourier Transforms Infrared spectroscopy (FTIR). The in vitro release studies were perfomed in the Franz difusion
cells using fosfate buffer pH 7,4 as mean of difusion. Results and Discussion: The nanoparticles exhibited
spherical shape and positive zeta potential values ranging about +25 mV. The dichloromethane/ethanol (50:50 v/v)
was established to produce the smallest particles (~150nm) and with drug loading efficiency ranging of 36% to
86%. The selected formulations demonstrated physicochemical stability over than six weeks. In addition, the
sialic acid or cholesterol decoration of particles changed substantially the EE% and drug release rate. Conclusion:
Thus, the emulsion-solvent evaporation method was efficiently optimized to produce stable BNZ-loaded PMMA
nanoparticles. The particles successfully decorated with sialic acid or cholesterol affected the drug release
properties, suggesting to be an interesting and promisor drug delivery system for BNZ targeting to infected cells.
Financial Support
Acknowledgments
Capes/Brazil (Pessoa LCST, Santos-Silva AM, Sousa DL, dos Santos-Silva E, Caland LB)
271
PHYSICOCHEMICAL CHARACTERIZATION OF HYDROGELS CONTAINING
NANOCAPSULES FOR TOPICAL DELIVERY OF TIOCONAZOLE
Lara Colles de Oliva Araújo
Fernanda Cramer Flores
Roberta da Silva Rosso
Federal University of Porto Alegre
Introduction: Superficial mycoses are fungal infections that occur on skin and are caused by
dermatophyte fungi and some yeasts. The topical therapy is impaired by low residence time of antifungal at the
site of action and the variable permeability of such substances. The use of nanotechnology for topical drug
delivery, due to the advantages, like drug targeting, providing a control of drug release, has been extensive
explored. This work aimed to prepare hydrogels containing polymeric nanocapsules for tioconazole delivery.
Methods: Tioconazole-loaded anionic and cationic polymeric nanocapsules (1 mg/mL, TIO-NC and TIO-C-NC,
respectively) were prepared by interfacial deposition of pre-formed polymer. The hydrogels containing the
nanocapsules (HG-TIO-NC and HG-TIO-C-NC) were prepared using an anionic polymer. The semisolid
formulations were characterized according to their rheological properties, particle size, pH and drug content. The
evaluation of tioconazole release from those hydrogels and from a hydrogel containing a tioconazole solution (1
mg/g, HG-TIO) was conducted using a Franz cells. In addition scanning electronic microscopy (SEM) of
hydrogels was performed. Results and Discussion: The rheological behavior of HG-TIO-NC and HG-TIO-C-NC
followed a non-Newtonian flow with pseudoplastic characteristics. The nanocapsules dispersed in the semisolid
showed nanometric particle size and the hydrogel presented slightly acid pH, with drug content close to the
theoretical values. After 72 h, HG-TIO-C-NC delivered 27% of the drug, and HG-TIO-NC released 20%, while
HG-TIO released about 90% of the drug in 24 h. SEM analysis showed the presence of submicrometric particles
in the hydrogels, inferring the maintenance of the nanocapsules. Conclusion: This work showed the feasibility in
obtaining hydrogels containing cationic and anionic nanocapsules for tioconazole delivery. The semisolid showed
adequate physicochemical properties and control of drug release.
Financial Support
CAPES, CNPq and FAPERGS
Acknowledgments
The authors thank for the support.
272
MICROSTRUCTURAL INVESTIGATION OF A MICROEMULSION CONTAINING A
THIOPHENE DERIVATIVE
Yuri Basilio Gomes Patriota
Natan Emanuell de Sobral e Silva
Vandiara Martins Moreira
Amaro César Lima de Assis
Mysrayn Yargo de Freitas Araujo Reis
Thiophenes are an important class of heterocyclic compounds, specifically the 2-amino substituted,
having a wide variety of biological properties including antibacterial, antifungal, analgesic and anti-inflammatory
activity. 2-[(2,4-dichloro-benzylidene)-amino]-5,6-dihydro-4H-cyclopenta [b] thiophene-3-carbonitrile (5CN06) is a
drug with highly lipophilic and poor water solubility. Microemulsion (ME) is a colloidal drug carrier system,
thermodynamically stable, containing oil, water and surfactant, having a small droplet size (10-100nm). This study
aimed to investigate the microstructure of a ME and the physical state of 5CN06 incorporated into the ME. The
ME was selected from a pseudoternary phase diagram (PTPD) containing isopropyl myristate (IM) as oil phase,
Labrasol® (LAS) as surfactant and distilled water (W), obtained by titration method. Thus, 5CN06 (0.02% w/v)
was characterized and incorporated into ME by three methods and their microstructures were investigated by
differential scanning calorimetry (DSC) and X-ray diffraction (XRD). From the PTPD, it was selected a ME
formulation containing 5% IM, 35% LAS and 60% W (w/w). All the 5CN06-MEs obtained from different methods
were clear, single phase and translucent dispersions. In the 5CN06-MEs diffractograms no peaks representing
crystals of 5CN06 were presented, even though the 5CN06 alone exhibited peaks in XRD analysis, indicating a
crystalline state. This result suggests an amorphization and incorporation of the drug into ME droplets. Both the
5CN06-ME and blank-ME thermograms showed two peaks representing the melting and freezing of water near 0
and -25 ºC, respectively. ME systems that exhibit only thermal behavior of water present an oil-in-water (O/W)
microstructure. Thus, we suggest that our system present this microstructure and these results indicated that
5CN06 was incorporated and probably protected in O/W ME droplets.
Financial Support
CETENE
Acknowledgments
CAPES, CNPq, CETENE
273
POLYMERIC NANOPARTICLES CONTAINING STANDARDIZED EXTRACT OF Ilex
paraguariensis A. St.-Hil. (ERVA-MATE)
Larissa Probst dos Santos
Flavio Henrique Reginatto
Angela Machado de Campos
Introduction: Ilex paraguariensis A. St.-Hil. is a vegetal species that has a significant antioxidant
properties attributed to its high content of phenolic compounds, specially chlorogenic acid (CGA). The
nanoencapsulation of standardized extract (SE) could protect the polyphenol CGA (chemical marker) from
hydrolysis degradation, however its hydrophilic character (Log Ko/w = -1) difficult this process. With this purpose,
two sequential formulation studies were performed. Methods: Initially obtaining of nanoparticles by a solvent
emulsification-evaporation method based on a w/o/w double emulsion was evaluated considering the following
variables: type of polymer (polycaprolactone, PCL, or poly(lactic-co-glycolic)acid, PLGA), type and amount of
surfactant (poloxamer 188, PLU, or polyvinyl alcohol , PVA, 0,5, 0,75, 1,0 and 5,0 %, w/v) and aqueous internal
phase volume of the first emulsion (200 or 400 µl). In a second study, it was evaluated: amount of SE (20 or 30
mg) and aqueous external phase volume (96 or 196 ml). The responses analyzed were particle size (PS, dynamic
light scattering), polydispersity index (PDI), zeta potential (ZP, laser-doppler anemometry) and CGA encapsulation
efficiency (EE, ultrafiltration/centrifugation, HPLC). The statistical evaluation of the results was performed using
the software Prism® 5. Results and Discussion: Nanoparticles with particle size less than 200 nm, PDI less than
0.150, ZP around -18 mV without precipitates were obtained using PCL as polymer (50 or 100 mg), PLU as
surfactant (1.0%, w/v) and 200 µl of aqueous internal phase volume. An EE of CGA about 40% was obtained in
the formulation containing 30 mg of SE and 196 ml of aqueous external phase volume, which is considered high
for a hydrophilic compound. Conclusion: The formulations studies allowed obtaining PCL-nanoparticles able to
encapsulate the standardized extracts of Ilex paraguariensis A. St.-Hil. with adequate physical-chemical
characteristics and high EE of CGA.
Financial Support
FAPESC/CNPq (PRONEM)
274
WASTE MATERIAL OF PROPOLIS AND GELATIN MICROPARTICLES CONTAINING
ASCORBIC ACID: MORPHOLOGICAL ANALYSIS, SIZE DISTRIBUTION AND
POLYDISPERSITY INDEX
Lizziane Maria Belloto de Francisco
Hélen Cássia Rosseto
Lucas de Alcântara Sica de Toledo
Rafaela Said Santos
Pharmaceutical inputs can be obtained by various processes, generating often byproducts or wastes,
which are not of interest. Many of these can be investigated for use in the preparation of medicaments.
Furthermore, the particulate systems have allowed an increase in therapeutic efficacy of side effects of many
drugs. Propolis is a strongly adhesive substance of complex chemical composition, showing important biological
activities. During the preparation of its extract, there is the formation of byproduct, usually discarded. However,
this waste material of propolis (WP) has shown the ability to form particles. Accordingly, to evaluate the kinetics
of release of the particles was used as model drug the ascorbic acid (AA), an excellent antioxidant, that has
problems of stability and thus, microencapsulation can be a strategy to solve this problem. Therefore, this study
aimed to obtain and characterize microparticles using the WP containing AA. Particles were prepared by
dispersing the WP extract 50% (w/w), gelatin and AA in purified water. The preparation was dropped under
mechanical stirring cold. The final dispersions were dried in a minispray dryer. Morphological analysis was
performed by SEM. The determination of the size distribution of the microparticles was performed using as
parameter for measuring second the Feret diameter, which approximately 1000 particles were counted. From
these results of the polydispersity index was determined. Most of the microparticles prepared exhibited small
size, spherical shape, some with depressions in the surface and different levels of agglomeration. Moreover, the
particles showed mean diameter between 2.41(± 1.54) and 4.26 (± 1.70) µm, with polydispersity index ranging
from 0.857 to 1.20. The present results indicated that is possible to prepare microparticles by spray drying method
using the WP containing ascorbic acid. In this sense, the characteristics of particles suggest than them can be
evaluated for AA delivery.
Financial Support
The authors thank the funding agencies CAPES, INTC_if, CNPq, FINEP.
Acknowledgments
At the State University of Maringá (UEM) and the Postgraduate Program in Pharmaceutical Sciences.
275
DEVELOPMENT, CHARACTERIZATION, EFFICACY IN VITRO, IN VIVO AND
PHAMACOKINETICS OF SURFACE-MODIFIED QUININE-LOADED NANOCAPSULES
Luana Roberta Michels
Tamara Ramos Maciel
Graziela Gomes Scheuer
Eduarda Piegas Martini
Sandra Elisa Haas
Introduction: The increase of Plasmodium falciparum resistance difficult the treatment of malaria.
Quinine (QN) is second line drug for the therapeutic, but its use is limited by side effects. The coating of
nanocapsules (NC) have been studied with the purpose of improve the in vivo performance of drugs. The aim of
this study was to develop, characterize and evaluate the efficacy in vitro, in vivo and pharmacokinetics of QN
loaded-PCL NC with surface modified by P80, PEG or cationic NC (Eudragit). Methods: Suspensions were
prepared by nanoprecipitation method and characterized according to diameter, polydispersity, pH, zeta potential,
drug content entrapment efficiency, atomic force microscopy and QN partition coefficient (D) into P. berghei
infected RBC (red blood cells). Pharmacokinetics was evaluated after iv administration of free or QN-loaded P80,
PEG or Eudragit NC (20 mg/kg). Peter's 4-day suppressive test against P. berghei infection in mice was used to
evaluate the efficacy of all NC. All experiments using animals had protocols approved by the ethical committee of
UNIPAMPA (010/2013). Results and discussion: PEG-coated NC showed smaller particle sizes when compared
to other formulations. Positive zeta potential was demonstrated for all formulations with Eudragit. The D doubled
when the drug was nanoencapsulated compared with the free drug. The increase on Vdss was responsible for the
prolongation of circulation half-life time observed for QN-loaded P80 coated NC and QN-loaded Eudragit-NC. To
PEG-coated NC, the decrement on Cltot increased QN plasmatic half-life. On the efficacy in vivo, cationic NC
increased the survival rate 9 days compared to saline and 7 days in survival compared to the free QN.
Conclusion: These results demonstrate that it is possible to develop appropriate NC-QN and that the drug
incorporated in the suspension with cationic characteristics can alter the efficacy of QN presenting as a potential
alternative for the treatment of malaria.
Financial Support
CNPq and FAPERGS.
Acknowledgments
CNPq and FAPERGS.
Ethical approval
UNIPAMPA (Protocol 010/2013)
276
COSOLVENT INTERACTIONS WITH HYDROXYPROPYL-BETA-CYCLODEXTRIN FOR
MODIFING AQUEOUS SOLUBILITY AND DISSOLUTION RATE OF BENZNIDAZOLE
Lucas Azevedo do Nascimento
Undegraduate Course of Pharmacy, Federal University of Rio Grande do Norte, Natal, RN, Brazil
Polyanne Nunes de Melo Doro
Graduate Program on Development and Technological Innovation of Medicines, Federal University of Rio Grande do Norte, Natal, RN,
Brazil
Arnóbio Antônio da Silva Junior
Laboratory of Pharmaceutical Technology and Biotechnology (TECBIOFAR), Pharmacy Department, Federal University of Rio Grande do
Norte, Natal, RN, Brazil
Introduction: Chagas disease is a neglected disease caused mainly by the parasite Trypanosoma cruzi
that affects about 7 million people around the world. Benznidazole (BNZ) is the only one drug commercially
available in Brazil for its treatment, but the poor aqueous solubility limits the drug dissolution rate. The present
study investigated the influence of the interaction of cosolvent triethanolamine (TEA) with hydroxypropyl-betacyclodextrin (HPβCD) in the aqueous solubility of BNZ. Methods: The increment on BNZ solubility and the
stoichiometry of inclusion complexes were found by phase solubility diagram and Job's plot. The in vitro
dissolution performance and physicochemical aspects of the freeze-dried solid complexes were carefully
assessed. Results and Discussion: Inclusion complexes with 1:1 stoichiometry were formed with or without TEA
at the same molar ratio of CD. The aqueous solubility of drug enhanced about seven times. FTIR, XRD and SEM
images confirmed the formation of amorphous solid complexes. The interactions among the compounds due to
the self-assembling of supramolecular aggregates were also identified in solid phase. In addition, the third
compound TEA enhanced the in vitro drug dissolution rate, compared with that from binary complex. This fact
occurred due to the stabilization effect of TEA in the BNZ:HPβCD interactions. Conclusion: Thereby, a potential
new solid raw material containing BNZ:HPβCD:TEA was successfully purposed for possible use as solid
dosage form or liquid dispersion for the Chagas disease treatment.
Financial Support
Capes/Brazil (P.N. Melo), Capes (23038.007487/2011-91) and CNPq (481767/2012-6)
277
CATIONIC NANOEMULSION ENHANCES ZINC PHTHALOCYANINE SKIN PENETRATION
Luciana Facco Dalmolin
University of São Paulo
Renata Fonseca Vianna Lopez
Nanoemulsions are emulsified systems with size ranging from 50 to 200 nm that are effective for
solubilization of poorly water soluble drugs. Their small droplet size and large superficial area can facilitate the
skin penetration of dispersed drugs. The aim of this work was to develop an oil-in-water cationic nanoemulsion for
topical administration of photosensitizer zinc phthalocyanine (ZnPc) for further photodynamic therapy and
compare its in vitro penetration to an emulsion. Nanoemulsion composed of medium chain triglyceride (MCT), egg
phosphatidylcholine, N-(2,3-Dioleoyloxy-1-propyl)trimethylammonium methyl sulfate, polysorbate 80, poloxamer
and water was produced by ultrasonication. ZnPc was solubilized in MCT and both phases were warmed to 70°C,
vortex mixed and ultrasonicated. Emulsion was produced as described above without ultrasonication. Both
formulations contained 15,41±1,25 µg/mL. Characterization was done by size, polydispersity index (PDI) and zeta
potential. In vitro skin penetration studies were carried out using dermatomed pig's ear skin fixed on Franz
diffusion cells and nanoemulsion/emulsion containing ZnPc (15µg/mL) was applied topically on the exposed skin
area. After 12h, the stratum corneum, removed by 15 tape strips and the remaining skin were processed. ZnPc
was assayed by fluorescence spectroscopy. Nanoemulsion obtained has size, PDI and zeta potential of
189.6±4.8 nm, 0.205±0.01 and 45±2 mV, respectively. In vitro penetration of ZnPc into the stratum corneum after
12h from the nanoemulsion (855 ± 106 ng/cm2) was 6.3 times higher than the emulsion (136±85 ng/cm2). ZnPc
penetration into the viable epidermis from the nanoemulsion was 77±14 ng/cm2 while it was not detected using
the emulsion. Cationic nanoemulsion containing ZnPc was successfully developed. In vitro skin penetration study
confirmed that the nanometric size of the nanoemulsion droplets favored and enhanced passive penetration of
ZnPc into the skin.
Financial Support
CNPq grant #141122/2015-3 and FAPESP #2014/22451-7
Acknowledgments
CNPq, FAPESP, FCFRP-USP
278
In vitro RELEASE STUDY OF TIOCONAZOL FROM OF REDISPERSIBLE POWDERS
AND FROM OF THE VAGINAL SUPPOSITORIES CONTAINING OF DRUG ASSOCIATED
AT NANOSPHERES
LUCIANA FILIPPIN COSSETIN
Jéssica Fernanda Wolf
Jocelene Filippin Cossetin
Introduction: The development of nanostructured materials has provided better control of the release and
targeting of active substances to the treatment site. These carriers are an alternative for the development of
products administered by vaginal route to further the distribution and penetration of drugs and reduced time spent
on site. From this, the present work aimed to study the in vitro release of vaginal suppositories and redispersible
powders containing tioconazole associated with nanospheres. Methods: Nanospheres suspensions containing
tioconazole (3 mg/mL) were prepared by nanoprecipitation method and were fed into a spray dryer. The vaginal
suppositories (25 mg/suppository) are prepared by molding method. In vitro release studies were evaluated by the
dialysis bag method, by using ethanol:water and simulated vaginal fluid as a release medium at 37 °C. Results
and Discussion: A mathematical model used to describe the best release profile fitting was biexponential model
for the both formulations. The rate constants observed for the burst (K1) were 0.185 ± 0.02 h-1 (TIO-SV), 0.02 ±
0.0009 h-1 (TIO-SV-NSP) and 0.052 ± 0.02 h-1 (TIO-NSP) and the observed rate constants for the sustained
phase (K2) were 0.0084 ± 0,00 h-1, 0.010 ± 0.00 h-1 and 0.0163 ± 0.0408 for SV-TIO, SV-TIO-NSP and TIO-NSP,
respectively. From this, suppositories consisting of the nanospheres enabled greater control of the release of
tioconazole compared with suppositories containing the drug not associated. Conclusion: These lower releases
showed that polymeric nanostructured system revealed a considerably prolonged TIO release profile even after
incorporating in solid formulation. Thus formulations proposed in this work may constitute an alternative for the
administration by vaginal route tioconazole, better acceptability compared to the existing dosage forms (creams,
gels) enabling, by definition, a greater therapeutic effect of the drug.
Financial Support
CAPES
Acknowledgments
CAPES
279
DEVELOPMENT OF A LONG-DURATION BIOTELEMETRY GEL FOR ECG SIGNAL
DETECTION - PRELIMINARY STUDY
Luciria de Freitas Correa
1Joan Vernikos Aerospace Pharmacy Laboratory, Pontifical Catholic University, Porto Alegre, RS, Brazil.
Isabel Rocha
Physiology Institute, University of Lisbon, Lisbon, Portugal.
Thais Russomano
Microgravity Centre - School of Engineering, Pontifical Catholic University, Porto Alegre, RS, Brazil.
Temis Furlanetto Corte
School of Pharmacy, Pontifical Catholic University, Porto Alegre, RS, Brazil.
Júlio César Marques de Lima
Endrigo Carvalho
Marlise Araújo dos Santos.
Joan Vernikos Aerospace Pharmacy Laboratory, Pontifical Catholic University, Porto Alegre, RS, Brazil.
Introduction: The analysis of bioelectrical signals has contributed to a better physiological monitoring of
individuals. The products available on the market permit the acquisition of physiological signals for approximately
2h30min, as they are mainly developed for quick medical exams, like ECG performance. However, monitoring for
a longer time period is necessary in many cases, such as athletes during heavy exercise, human physiology
studies, and patients suffering from a range of different diseases. Objectives: To develop a new conductive gel
formulation that allows the acquisition of cardiovascular physiological signals for 12 hours. Methods: Six
conductive gel formulations were developed that were tested using the TEMIS system, which consists of a Tshirt with sensors connected to a mobile platform for data acquisition. The ECG signals were registered from
healthy volunteers, comparing the new formulations with water and a commercial gel (Lectromed). The tests were
performed on consecutive days during 12 hours during which the new gels, water and Lectromed gel were applied
using the TEMIS system and registered on a tachogram. The signals obtained were analyzed using the FisioSinal
program and the degree of reliability evaluated. Results and Discussion: Data analysis showed that gel number 1
presented a longer durability by maintaining the electrical properties for approximately 4h30min, which proved to
be more effective than the commercially available gel. It was verified that the type, polymer concentration, and
humectant employed influenced the conduction time of the gel. Furthermore, the findings demonstrated that
silicone negatively interfered in the contact time. Conclusion: This preliminary study showed that the gel
formulation 1 presented an improvement in relation to the commercial gel against which it was tested. However,
the durability of 12h expected fromthe new gel formulations was not obtained, motivating continuation of this
research.
Ethical approval
334/14
280
INNOVATIVE MUCOADHESIVE FORMULATION CONTAINING IMIQUIMOD-LOADED
NANOCAPSULES
Paula dos Santos Chaves
Flávia Pires Peña
Introduction: Prior studies have shown the efficacy of imiquimod in the treatment of human
papillomavirus (HPV). However, some adverse effects are reported for women using the commercial formulation.
This could be avoided with the use of nanotechnology. There is also a need of adhesive systems so that the drug
can remain more time in the mucosa. The use of polymeric nanocapsules can help to increase the drug
permanence in the mucosa, leading to an improvement of its formulation effect. Objective: To develop
nanotechnology formulations containing imiquimod involving polymeric nanocapsules with and without the cationic
polymer chitosan. The formulations performance are compared in order to increase the drug mucoadhesion and
penetration in the vaginal tissue pursuing the most promising system for treatment of HPV. Materials and
Methods: Two types of nanocapsules were developed by nanoprecipitation: NCimiq (nanocapsules containing
imiquimod) and NCimiq-chit (nanocapsules containing imiquimod with chitosan on the surface). Mucoadhesion
was determined by texture analysis. In vitro drug permeability profiles from nanocapsules and free drug (IMIQfree)
were evaluated using fresh vaginal mucosa and modified manual Franz diffusion cell. Results and Discussion:
The work of 22.3 mN.mm (NCimiq) and 17.15 mN.mm (NCimiq-chit) was necessary to detach the formulations
from the vaginal mucosa (not representing statistical differences between them; i.e. p>0.05). This result confirmed
the mucoadhesive properties of nanocapsules. In vitro drug permeability showed that imiquimod is able to cross
sublingual mucosa and that the nanoencapsulation controlled the permeability, resulting in less adverse effects.
Conclusions: Nanoencapsulation presents adequate mucoadhesion on the vaginal mucosa leading to an
increasing effect in the treatment of HPV. This formulation controlls imiquimod permeation and, consequently, can
be alternative for the treatment of adenocarcinoma.
Financial Support
CNPq/Brazil and CAPES/Brazil
Acknowledgments
PPGCF
281
FLAVONE NANOEMULSION SYSTEM: DEVELOPMENT AND CHARACTERIZATION
JOANDRA MAÍSA DA SILVA LEITE
TÚLIO CHAVES MENDES
BRUNA PEREIRA DA SILVA
LARISSA PEREIRA ALVES
ANA CLÁUDIA GONÇALVES DOS SANTOS
AIRLLA LAANA DE MEDEIROS CAVALCANTI
ANNA EMANUELA MEDEIROS DE BRITO
CAMILA DE OLIVEIRA MELO
BOLÍVAR PONCIANO DE LIMA DAMASCENO
Nanoemulsions are transparent, isotropic and kinetically stable systems. They are also versatile
systems able to carry lipophilic, amphiphilic and hydrophilic drugs and to control its release, thus reducing the
possibility of adverse effects. Thus, the aim of this work was the development and characterization of a
nanoemulsion (NE) system for the incorporation of a flavonoidic derivative with antimicrobial potential. The
systems were formulated from the study of the hydrophilic lipophilic balance (HLB) oil (isopropyl myristate) and
then defined as surfactants Span 80 and Kolliphor HS15 (9: 1) by HLB similarity. The pseudoternary phase
diagram (PTPD) were constructed and the point that showed transparent and compatible with the NE O/A was
selected and characterized by the pH analysis, polarized light microscopy and droplet size, polidispersion index
and Zeta potential by DLS. The formulation proportions obtained has 10.8% oil, 25.87% surfactants and 63.33%
water, typical of O/A systems. The study revealed that the formulation has slightly acidic character (pH =
6.78±0.028). In the photomicrographs derived from the polarized light microscopy, it was observed isotropy
characteristic present in the NE. Through analysis by light scattering it is possible to classify the formulation as
polymodal, with droplet size of 0.01593 µm and zeta potential of -72,1 mV. It can be concluded that the selected
formulation shows potential for incorporation of lipophilic drugs, and demonstrate consistent properties for topical
administration.
Financial Support
CETENE
Acknowledgments
CAPES, CNPQ.
282
ANTIOXIDANT ACTIVITY, LIPID PEROXIDATION AND ANTIPROLIFERATIVE EFFECT OF
CO-ENCAPSULATED QUERCETIN AND RESVERATROL INTO LIPOSOMES ON HL-60
CELLS
Marcela Araújo Pereira
Daniel Charles dos Santos Macêdo
Rafaela Siqueira Ferraz Carvalho
Paulo Euzebio Cabral Filho
Jaciana dos Santos Aguiar
Teresinha Gonçalves Silva
Adriana Fontes
Universidade Federal Rural de Pernambuco (UFRPE)
Pabyton Gonçalves Cadena
Nereide Stella Santos Magalhães
Phytochemicals are natural compounds found in plants that present numerous pharmacological
activities. Quercetin(QUE)and resveratrol(RES)have been directly associated with anticancer effects due to its
antioxidant activity. Their use in therapy is still limited because of the low aqueous solubility, which leads to a low
bioavailability. An approach able of overcoming these limitations is their incorporation in drug controlled release
systems such as liposomes. The aim of the present study is to evaluate the effectiveness of QUE and RES
nanoencapsulated into elastic liposomes (QUE/RES-Lipo) by antioxidant activity, content of lipid peroxidation by
TBARS (thiobarbituric acid reactive species) method, antiproliferative activity, determined by MTT (3-(4.5dimetiltiazol-2il) -2.5-diphenyl tetrazoline bromide) assay and elucidation of the cell death mechanism with dyes
annexin and propidium iodide marking by flow cytometry. The antioxidant activity, determined using ABTS [2.2'azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) salt diammonium] method, showed that,
at the
same
concentration, QUE had greater activity (TEAC 1091.93±15.4µM) comparing with RES (TEAC 867.98±31.75µM).
Moreover, the encapsulation of QUE/RES into liposomes increased antioxidant activity in 12% compared with
free QUE or RES and reduced the content of lipid peroxidation in 56%(TBARS).In addition, QUE/RES-Lipo
presented the best antiproliferative activity, reducing cell viability in 81% with a reduction of 25% on IC50 (with
IC50 values of 144.23 µM for QUE/RES and with 85.43 µM for QUE/RES-Lipo). As mechanism of cell death, it
was verified that QUE and RES act by apoptosis with 81.8 % of Annexin-V marcation for QUE/RES-Lipo, with an
increase of 60% in relation to free drugs. Based on the results, we can conclude that the liposomal formulation
containing QUE and RES nanoencapsulated presented a good activity against HL-60 cell line, leading to future
studies aiming to improve their efficacy.
Financial Support
CNPq and FACEPE
Acknowledgments
LIKA, UFPE
283
VALIDATION OF AN HPLC-UV METHOD FOR ANALYSIS OF CURCUMIN C3 COMPLEX
Marcela Brito Oliveira
State University of Maringá - UEM
Marcela Brito Oliveira
Introduction: Curcumin (CUR) is a compound extracted from Curcuma longa Linn, which belongs to the
Zingiberaceae family. CUR is present in curry and turmeric (very common in typical foods from India and China),
which gives a yellow-orange color of striking odor. It has been extensively studied, because has activity against
various types of diseases including bladder cancer. The aim was to validate an analytical methodology to
evaluate the CUR. Methods: CUR standard (purity ≥ 98%) and a donated sample of CUR C3 complex (Sabinsa
Corporation) in HPLC with UV-visible detector and manual injector was used. The analysis conditions were:
isocratic condition; RP18 column (5μm, 25 cm x 4.6 mm), at 25 °C, wavelength of 425 nm, total run time of12
min, flow rate of 2.0 mL/min, and injection volume of 20 μL. The mobile phase consisted of acetonitrile and acetic
acid solution 1.5% (V/V) in the ratio 42:58 (V/V). Analytical curve was obtained from CUR stock solution (1.0
mg/mL) prepared in methanol in order to obtain dilutions from 4-200 µg/mL. The following parameters were
evaluated: selectivity; limit of detection (LD) and limit of quantification (LQ); linearity; precision, accuracy and
robustness. Results and Discussion: The method displayed selectivity for detecting the drug and showed linearity
between 6-200 µg/mL (except 4 and 32 µg.mL-1). The curve did not present lack of fit and R = 0.9985. The LD
and LQ were 2.69 e 8.14 μg/mL, respectively. The precision was satisfactory with RSD values below 5% and
accuracy between 98.38 and 99.73%, concerning the repeatability of the method intra- and inter-day respectively.
The robustness showed statistically significant differences (p<0.05) between the variation of the wavelength and
flow compared to standard conditions. Conclusions: The results showed that the analytical methodology is valid
to quantify CUR. We thereafter shall be quantified the curcumin content present in the nanoparticles.
Financial Support
CNPq, CAPES and FINEP (Brazil).
Acknowledgments
Sabinsa Corporation for having donated the sample of curcumin C3 complex.
284
OPTIMIZATION OF CURCUMIN LOADED LIPID NANOPARTICLES FORMULATED USING
SOLID DISPERSION
Franciely Cristina Corrêa das Neves Lopes
Rodrigo Villares Portugal
Brazilian Nanotechnology National Laboratory
Marcelo Alexandre de Farias
Patricia Severino
Universidade Tiradentes (UNIT)
Cecília Torquetti de Barros
Introduction: Solid lipid nanoparticles (SLN) are drug carriers possessing advantages with respect to
stability, drug release profile and biocompatibility. Curcumin (Cur) is a liposoluble polyphenolic compound that
possesses great therapeutic potential. Its clinical application is, however, limited by poorly water solubility. One
key strategy for improving the solubility and bioavailability of CUR is solid dispersion technology. This work aimed
at improving the solubility and stability of Cur by the preparation of fusion binary solid dispersions (SD) and loaded
in SLN. Methods: The CUR-SD were prepared by microwave method using polyvinylpyrrolidone K30 (PVP K30)
as carrier. CURSD-SLN were formed by lipid recrystallization. The resulting hot o/w nanoemulsion was dumped
into ice cold distilled water (2°C), quickly under high speed stirring (12,000 rpm/5 min). The SD were
characterized by solubility study, FTIR spectroscopy, XR diffraction, DSC and SEM. The CURSD-SLN were
characterized by particle size (PS), polydispersity index (PI), zeta potential (ZP), encapsulation efficiency (EE),
transmission electron microscope (TEM). Results and Discussion. The solubility of CUR-SD increased ~1,000x.
The absence of peaks characteristic reflection in the CUR-SD suggest reducing the crystallinity of curcumin,
XRD data were in good agreement with the results depicted by DSC and MEV photography. The PS, PI, ZP, EE
of the SLN optimised formulation (CURSD-SLN) were 140 nm, 0.23, −19.9 mV, and 90%, respectively. TEM
showed the presence of nanoparticles with two different structures, one is multilamellar and the other with a
dense core formed by the polymer which extend in loops, in the shape of petals of a flower formed by the lipid
bilayer. The release profile, in vitro, demonstrated curcumin-loaded SLN can control Cur release. Conclusions:
Curcumin-loaded solid lipid nanoparticles may be a promising drug delivery system to control drug release and
improve bioavailability.
Financial Support
FINEP. CNPq
Acknowledgments
285
PECTIN/PROPOLIS BYPRODUCT MICROPARTICLES CONTAINING L-ALANYL-LGLUTAMINE: DEVELOPMENT AND EVALUATION OF THE IMMUNOSTIMULANT
ACTIVITY IN NEUTROPHILS
Mônica Villa Nova
Bianca Altrão Ratti
Natália de Carvalho Scharf Santana
Sueli de Oliveira Silva Lautenschlager
Introduction: Catabolic states such as cancer are associated with immunosuppression. L-alanyl-Lglutamine (AGP) enhances immune function by acting as a substrate for lymphocytes and neutrophils. Moreover,
propolis also has immunomodulatory activity. Because of the fast metabolism of AGP after oral administration,
the aim of this work was to prepare microparticles (MP) containing AGP using pectin and the byproduct of
propolis extraction as matrices of encapsulation and evaluate the morphology, encapsulation efficiency (EE) and
the ability of these MP to induce the production of hypochlorous acid (HOCl) by human neutrophils. Methods: An
extractive solution (30%, w/w) of propolis was prepared using 96 °GL ethanol and turboextraction and the
resultant byproduct was used to prepare a 50% (w/w) hydroethanolic extract of byproduct (HEB) with 96 °GL
ethanol. Afterwards, the HEB and pectin were used to prepare blank and AGP-loaded particles by spray drying
technique. Scanning electron microscopy was used for morphological analysis and the EE was determined by a
validated HPLC method (C18 column (Supelco®), mobile phase composed of acetonitrile/0.1% aqueous
trifluoracetic acid solution (57.5:42.5%), flow rate of 0.8 mL/min, UV-VIS detector at 210 nm and temperature of
30 ºC). The formation of HOCl was followed by spectrophotometry based on the formation of taurine chloramine
resulting by the reaction of HOCl with taurine. Results and Discussion: Small spherical MP with smooth surface
were obtained and an EE of 69.39 ± 3.10% was achieved. The blank MP were able to increase the production of
HOCl by neutrophils (p<0.001) and the results with the AGP-loaded particles was comparable to the positive
control (p<0.0001). Conclusion: Pectin and byproduct of propolis showed suitable as materials to encapsulate the
dipeptide with excellent results of EE and great morphological characteristics. The particulate systems showed
promising results in the immunostimulation of neutrophils.
Financial Support
CAPES, CNPq, and FINEP
Acknowledgments
286
DEVELOPMENT AND CHARACTERIZATION OF HYBRID LIPID-POLYETHYLENIMINE
(PEI) NANOCARRIERS FOR DELIVERY TOPICAL SIRNA: IN VITRO STUDIES
Margarete Moreno de Araujo
Faculdade de Ciencias Farmaceuticas de Ribeirão Preto - USP
Priscyla Daniely Marcato Gaspari
Graduate Program in Pharmaceutical Nanotechnology, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo,
Ribeirão Preto, São Paulo, Brazil
Maria Vitoria Lopes Badra Bentley
Graduate Program in Pharmaceutical Sciences, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão
Preto, São Paulo, Brazil
Lipid-polymeric nanocarriers are the new generation of nanoparticulate that have been used as siRNA
carrier system and are attracting attention as novel colloidal drug carrier for topical use. The application of
nanoparticles on a sub-cellular level still needs in depth studies of their biocompatibility. However, a complete
characterization of the particles under the physiological conditions is still lacking. The goal of this study was to
develop of hybrid lipid-polyethylenimine nanocarriers (NC) as topical delivery systems for siRNA and to determine
the ability of the NC complexes to internalize the siRNA molecules and visualize the cellular localization of the
complexes. The composition and method of production of NC, using Compritol 888 ATO®, Polaxamer 188 and
PEI (0.15 and 0.25%), were optimized by turrax homogenizer. This delivery systems has complexed the siRNA
and was characterized (by Zeta potential, Dynamic Light Scattering-DLS, Differental Scanning Calorimetry-DSC
and Stability Studies). The nanoparticles had a mean size of 210 nm and an average zeta potential of 15 mV and
remained stable for 90 days at 4°C; it was demonstrated by DSC, the surfactante and the lipid interacts with PEI
on the surface of the nanoparticles. In vitro studies, it was worked with humans keratinocytes (HaCat cells), the
nanoparticles showed high viability by the resazurin reduction assay and high transfection evidenced by flow
cytometry (about 75% of transfected cells) and by confocal microscopy. The penetration NC complexed with
siRNA-FAM into the skin was assessed using an in vitro model of porcine ear skin, was observed the
nanoparticles did not reach the epidermis by fluorescence microscopy. Based on these results, it can be
concluded that the developed formulations are promising delivery systems for topical administration of siRNA for
the treatment of skin diseases in gene therapy.
Financial Support
FAPESP, CAPES e CNPq
287
SOLID LIPID NANOPARTICLES CONTAINING HESPERIDIN: OBTAINING AND
CHARACTERIZATION
Mariana Correia Kossatz
Department of Pharmaceutical Sciences, State University of Ponta Grossa, Ponta Grossa, PR, Brazil
Clarissa Elize Lopes
Andressa Novatski
Department of Physics, State University of Ponta Grossa, Ponta Grossa, PR, Brazil
Lidiane Vizioli Castro
Department of Physics, State University of Maringá, Maringá, PR, Brazil
Priscileila Colerato Ferrari
Introduction: Hesperidin, a bioflavonoids glycoside, has a wide variety of pharmacological applications,
in particular, antioxidant activity, as protective effects in neurodegenerative diseases. Solid lipid nanoparticles
(SLN) carrying hesperidin are a promising tool to increase their bioavailability by target the drug delivery. This
work consists in the development of SLN carrying hesperidin and their physicochemical characterization.
Methods: SLN were prepared by hot microemulsion technique constituted by an lipid phase composed by stearic
acid (SA) or cetostearyl alcohol (CA) and polyvinyl alcohol (PVA) or polysorbate 80 (PS), as surfactant, added in
aqueous phase. The mean diameter, polydispersity index and potential zeta were measured by Photon Correlation
Spectroscopy (Zetasizer Nanoserier) and SLN, free drug, lipids and physical mixtures are evaluated by Raman
Spectroscopy with Fourier Transformed (FT-Raman) (Nd:YAG laser, 1084 nm) and Photoacoustic Spectroscopy
(Home Made Setup). Results and Discussion: SLN presented mean diameter varying from 177 to 260 nm and
polydispersity index lower than 0.4 to all samples; however SA and PS samples showing more particle size
homogeneity (values < 0.3) than samples composed by CA and PVA. Negative zeta potential was obtained, due
to lipids properties, and values between -18.5 and -26.3 mV, which indicate the electrical stability of SLN to avoid
nanoparticles aggregation. FT-Raman spectrums showed some peaks displacement comparing nanoparticles and
physical mixture evidencing the drug encapsulation. Photoacoustic spectrums also confirm the drug encapsulation
due to the peaks of drug were more evident in 284 and 370 nm, regions in which the drug absorbs. Conclusions:
SLN containing hesperidin were successfully obtained and their constituted by SA and PS showed better
characteristics. Physicochemical evaluation confirmed the drug encapsulation.
Financial Support
Fundação Araucária
Acknowledgments
Profa. Dra. Josiane Padilha (DEFAR-UEPG), C-LABMU-UEPG, COMCAP-UEM
288
THE INFLUENCE OF HYPERICIN IN A BINARY POLYMERIC SYSTEM: CONTINUOUS
SHEAR RHEOLOGY
Mariana Volpato Junqueira
Fernanda Belincanta Borghi-Pangoni
Hypericin (Hyp) is a natural photoactive pigment originated from the plants of the Hypericum genus,
especially in Hypericum perforatum species, commonly known as St. John's wort. Many studies have been
conducted with in vivo and in vitro assays and demonstrated the antitumor activity from Hyp using Photodynamic
Therapy (PDT). Polymeric systems composed by Poloxamer 407 (Polox), Carbopol 934P (Carb) and Hyp can be
administered in order to benefit the colorectal cancer treatment with PDT association. The formulation with and in
absence of Hyp were prepared with polymeric dispersion of Carb (0.15%, w/w) in distilled water with following
addition of Polox (20%, w/w), triethanolamine as the neutralizing agent, and 0.01% (w/w) of Hyp. Continuous
shear rheograms were obtained using a rheometer (MARS II, Thermo-Haake) at 25 ºC and 37 ºC (n=5). The
obtained results were analysed using the Ostwald-the-Waele equation (Power Law). The influence of temperature
(T°C) and the presence of Hyp on the flow properties were statistically evaluated using ANOVA (p<0.05).The
formulations presented shear thinning flow behavior, being classified as plastic fluids at both T°C. The increase of
T°C resulted in an increase of shear stress and hysteresis area. The presence of Hyp decreased the n value (flow
index) from 0.716 to 0.18 at 25 °C, and from 0.955 to 0.19 at 37 °C. However, the consistency index (κ) value
increased from 3.701 to 179.720 at 25 °C, and from 0.425 to 273.800 at 37 °C. The same results were observed
with the increase of T°C (from 179.720 to 273.800, in the presence of Hyp). All these results displayed that the
formulation becomes more fluid at 25 °C, but at 37 °C (at the intestine region) the systems has restructured and
become thicker. These characteristics allow the system to remain in the intestine and the PDT can be performed.
Financial Support
CAPES, CNPq, Fundação Araucária, FINEP, UEM.
289
INVESTIGATION OF THE WOUND HEALING ACTIVITY OF COUMESTROL IN
DIFFERENT FORMS: FREE OR ASSOCIATED WITH HYDROXYPROPYLΒ
CYCLODEXTRIN
Marília Garcez Corrêa da Silva
Health Sciences Post-Graduate Program, University of Rio Grande, Rio Grande do Sul, RS, Brazil
Barbara Elisabeth Kummer Machado
Pharmaceutical Sciences Post-Graduate Program, University of Rio Grande do Sul, Porto Alegre, RS, Brazil
Michelle Maidana Altenhofen da Silva
Lidiane Dal Bosco
Magno Marques
Comparative Animal Physiology Post-Graduate Program, University of Rio Grande, Rio Grande do Sul, RS, Brazil
Ana Paula Horn
Comparative Animal Physiology Post-Graduate Program, University of Rio Grande, Rio Grande do Sul, RS, Brazil
Cristiana Lima Dora
Valquiria Linck Bassani
Pharmaceutical Sciences Post-Graduate Program, University of Rio Grande do Sul, Porto Alegre, RS, Brazil
Introduction: Chronic wounds and severe burns lead to exposure of the body and can compromise the
spontaneous recovery of the skin. Therefore, the discovery of new formulations capable of enhancing tissue
healing can improve the patient quality of life. Coumestrol (COU) is a potent antioxidant with estrogenic activity.
However its use in pharmaceutical formulations is limited by their low water solubility. In this context, the
hydroxypropylβcyclodextrin (CD) appears as an excipient capable to increase COU solubility. The aim of this
study is to evaluate the wound healing activity of coumestrol in different forms: free or associated with CD by the
spray drying method (COU: CD-SD), incorporated in hypromellose hydrogel (HPMC) by using Wistar rat
cutaneous excision wound model. Methods: The animals were divided in 4 groups: Dersani® (positive control),
HPMC hydrogel (negative control), coumestrol associated with CD incorporated into the hydrogel of HPMC
(HPMC + COU: CD-SD) and coumestrol incorporated into HPMC hydrogel. Skin samples were collected on 2th,
7th and 12th days after wounding. The wound contraction and histological analysis were evaluated. The
experimental procedure was approved by the Institutional Animal Care and Use Committee (CEUA-FURG),
protocol code 23116.003350/2015-11. Results and Discussion: The results from wound contraction over 12 days
of treatment indicated that the treatment carried out with HPMC + COU: CD-SD was similar in efficacy as
compared with the positive control (Dersani®), but more efficient, once 50% of the wound closure was achieved
in less time. Furthermore, histological analysis indicated that both treatments showed adequate tissue
organization. Conclusion: The association of coumestrol/cyclodextrin seems to accelerate wound healing,
probably due to their involvement with the dermal remodeling.
Financial Support
CNPq/Brazil and master scholarship from CAPES/Brazil.
Ethical approval
The experimental procedure was approved by the Institutional Animal Care and Use Committee (CEUA-FURG), protocol code 23116. 003350/2015-11
290
QUALITY EVALUATION OF COMPOUNDED SEMISOLID FORMULATIONS CONTAINING
NIMESULIDE
MARINA GRABOSKI
UNIVERSITY OF PASSO FUNDO
MIRIAM TEREZINHA KNORST
UNIVERSITY OF PASSO FUNDO
Topical administration of the nonsteroidal anti-inflammatory drug (NSAID) nimesulide may be a safe and
effective alternative to oral and rectal routes. Topical administration of NSAIDs offers the advantage of locally
enhance drug delivery to affected tissues. Therefore, in the case of regional inflammatory disorders such as
muscle pain and osteoarthritis, the topical application of NSAIDs has been preferred. The purpose of this study
was to evaluate the quality of semisolid formulations containing nimesulide compounded in pharmacies of the
state of Rio Grande do Sul. Five samples of semisolid formulations, containing 2.0 % of nimesulide, were
obtained from different pharmacies in Rio Grande do Sul. The samples, assigned as A, B, C, D and E, were
evaluated in relation to labelling (based on the Resolution RDC 67/2007 of ANVISA/MS), organoleptic
characteristics, pH and nimesulide content, according to the Brazilian Pharmacopoeia. The results show that only
three of the five evaluated formulations (Products A, B and E) fulfilled all the requirements for labelling. All
semisolid formulations showed a homogeneous light yellow coloration and presented pH values compatible with
the skin. The products A, B, C and D were satisfactorily conditioned in opaque plastic tubes. The nimesulide
content was within the specification only in the A and C semisolid formulations (between 90 and 110%). Only
product A fulfilled all the official specifications. The results show that the Good Practices of Manipulation in
Pharmacy are not being fulfilled by some pharmacies of the state of Rio Grande do Sul, since products without
the required quality are being marketed.
Financial Support
-
Acknowledgments
-
Ethical approval
-
291
POLYMERIC FILM COMPOSITION: A STRATEGY TO MODULATE THE AMPHOTERICIN
B RELEASE
Marjorie Caroline Liberato Cavalcanti Freire
Thiago Gomes Silva
Francisco Alexandrino Júnior
Paulo Henrique de Souza Picciani
Kattya Gyselle de Holanda e Silva
Julieta Genre
Eryvaldo Sócrates Tabosa do Egito
Introduction: Drug delivery systems are able to control active ingredients release, enhancing their
physicochemical properties and bioavailability. Understanding the factors that influence drug release will
determine the effectiveness of delivery system and is mandatory for the development of new and effective
therapeutics. The aim of this study was to assess Amphotericin B (AmB) release kinetics profiles from film
polymeric systems with different composition, hydrophilic (PVA) or hydrophobic (PLA), and also to correlate these
profiles with thermodynamic parameters using mathematical approaches. Methods: Films were produced by the
casting technique and evaluated at distinct temperatures for AmB encapsulation efficiency, porosity by Scanning
Electronic Microscopy (SEM), superficial wettability and AmB release. At the end, a mathematical modeling was
applied to evaluate release profiles. Then, the activation energy of the systems was determined using the
Arrhenius equation, and enthalpy, entropy and Gibbs free energy were calculated through the Eyring equation.
Results and Discussion: SEM analysis of PVA and PLA films showed absence of pores and no phase separation
between the polymer and the drug. PVA films showed greater release control, with maximum percentage of drug
released after 120 hours (84.32 ± 0.70%). PLA films showed low drug release profile possibly due to the
hydrophobicity of the polymer, verified by superficial wettability, presenting maximum AmB release of 4.12 ±
0.35% after 72 hours. PLA and PVA systems fitted to Higuchi's model suggesting that the major mechanism of
AmB release from these systems was based on Fickian diffusion. Thermodynamic parameters corroborated the
release kinetics data, revealing that AmB release is an endothermic and non-spontaneous process. Conclusion:
The composition of polymeric films can be used as a strategy to enhance AmB controlled release and
thermodynamic parameters can be used to explain its kinetic release profile.
Financial Support
CAPES, CNPq, PROPESQ, PPgCF, PPgCSA and UFRN
292
EVALUATION OF DISSOLUTION PROFILE OF METOPROLOL SUCCINATE
FORMULATIONS PRODUCED BY HOT MELT EXTRUSION
Michele Georges Issa
Natalia Vieira de Souza
Vinícius Texeira da Silva
Jéssika da Silva Lopes
Marcelo Dutra Duque
Muqdaq Alhijjaj
University of East Anglia
Sheng Qi
University of East Anglia
Humberto Gomes Ferraz
Hot melt extrusion (HME) is an interesting technique for pharmaceutical companies, as it makes
possible to obtain drug products faster than other methods and in few steps. Furthermore, depending on
excipients, mainly polymers, different drug delivery systems can be obtained. Studies show that some
hydrophobic polymers commonly used in matrix systems present great potential to control the release of highly
soluble drugs when submitted to HME. The objective of this study was to produce formulations containing
metoprolol succinate (MS) and Kollidon SR® by HME and evaluate drug dissolution. The formulations were
obtained according to a full factorial two level experimental design. Independent variables were percent of drug
into formulation (10% and 30%), process temperature (100 °C and 130 °C) and extruded size (< 2 mm and 5 to 7
mm). The obtained formulations were characterized by differential scanning calorimetry (DSC) and packed into
capsules to dissolution profile evaluation using U.S. Pharmacopeia apparatus 2 at 50 rpm, 500 mL of phosphate
buffer pH 6.8 during 20 hours. DSC curves showed suppression of the endothermic event, correspondent to
melting of the drug, in the formulations containing 10% of MS, which means drug solubilized in the polymer or
amorphous state. For the formulations containing 30% of MS, the melting event was observed, but less evident,
indicating that part of the drug in crystalline form was dispersed in the polymer. According to dissolution results,
extruded size and process temperature are directly related to the potential of drug release and formulations
containing 30% of MS, produced at 100 °C with extruded size between 5 and 7 mm, showed an interesting profile
modulation.
Financial Support
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES/Brazil)
293
INVESTIGATION OF THE ANTITUMOR ACTIVITY OF LONG-CIRCULATING AND pHSENSITIVE LIPOSOMES CONTAINING DOXORUBICIN IN EXPERIMENTAL ANIMAL
MODEL
Mônica Cristina Oliveira
Universidade Federal de Minas Gerais
Andre Luís Branco de Barros
[email protected]
Sávia Caldeira de Araújo Lopes
[email protected]
Elaine Amaral Leite
[email protected]
Geovanni Dantas Cassali
[email protected]
Diego dos Santos Ferreira
[email protected]
Fernanda Alves Boratto
[email protected]
Introduction: The encapsulation of doxorubicin (DXR) in long-circulating liposomes helped to reduce the
incidence of DXR cardiac toxicity, which led to the approval by FDA in 1995 of the Doxil®. Despite its
advantages, generally long-circulating liposomes tend not to be able to merge with the endosomal membrane upon
cell internalization. Therefore, pH-sensitive liposomal formulations containing anticancer drugs have been
developed to take advantage of the endosomal acidification, which leads to destabilization of the liposomal
membranes followed by release of its internal content.Objective: To develop and characterize physicochemically a
long-circulating and pH-sensitive liposomal formulation containing DXR (SpHL-DXR) and evaluate the cytotoxicity
against the murine breast tumor cell line 4T1 and the antitumor activity in a breast cancer animal model.
Methodology:SpHL-DXR was prepared by thin film hydration method and it was characterized by average
diameter, polydispersity index (IP), zeta potential and encapsulation percentage. Cytotoxicity against 4T1 cell line
was evaluated by MTT assay. For antitumor activity studies, breast tumor-bearing mice were intravenously
treated with HEPES-buffered saline pH 7.4, blank liposomes, free DXR, liposomal formulation similar to Doxil® or
SpHL-DXR (4 mg of DXR/Kg) every two days, in a total of four administrations.The tumor volume was evaluated
during the treatment period. Results and Discussion:The average diameter of SpHL-DXR was below 200 nm with
IP no higher than 0.2 and zeta potential close to neutrality. Encapsulation percentage was equal to 100%. IC50
obtained for SpHL-DXR treatment was equal to 1563 nM and for free DXR was equal to 2413 nM. Considering
the tumor volume analyses, SpHL-DXR was able to stabilize tumor growth in contrast to free DXR.
Conclusions:SpHL-DXR showed physicochemical characteristics that allowed its application for in vivo studies
and better antitumor activity when compared with free DOX.
Financial Support
Fundação de Amparo a Pesquisa do Estado de Minas Gerais
Acknowledgments
The authors would like to thank Fundação de Amparo à Pesquisa do Estado de Minas Gerais - FAPEMIG for the financial support (REDE-40/11). The authors
also thank FAPEMIG for supporting Fernanda Alves Boratto with a scholarship.
Ethical approval
The in vivo studies were conducted under approval of the Ethics Committee on Animal Use (CEUA) of the UFMG - Protocol 41/2015.
294
DEVELOPMENT OF SOLID DISPERSIONS SYSTEMS FOR PH DEPENDENT RELEASE
OF TAMOXIFEN
MONICA FELTS DE LA ROCA SOARES
Dayanne Tomaz Casimiro da Silva
Jose Lamartine Soares Sobrinho
Tamoxifen (TMX) is classified as class II in the Biopharmaceutical Classification System and has low
aqueous solubility, hindering its absorption after oral administration. As solution, solid dispersion systems are an
interesting approach to enhance solubility and oral bioavailability. The aim of this work was to obtain and
characterize solid dispersion systems by x-Ray diffraction and to evaluate drug delivery systems. Solid
dispersions were obtained by the solvent evaporation method, using the Eudragit® polymers L100 and RL100 and
TMX at three different concentrations. It was performed X-ray Diffraction and saturation concentration in the
means of pH 1.2, pH 7.4 and pH 7.4 with Tween80 (1%w/v). The drug delivery approach was performed under
non-sink conditions and were evaluated in UV-vis spectrophotometer at 274 nm and 307 nm. The dose was
calculated by Sink Index (SI). TMX presented a crystalline pattern which was not observed at solid dispersion
systems, when several peaks were absents. At a 0.33 SI and pH 1.2, TMX displayed a prompt dissolution and at
pH 7.4 there is a supersaturating rate. For the SD10%, SD20% and SD30% there was a slower release, in 2
hours of dissolution, for pH 1.2 of 6.0µg/mL (5%), 12.0µg/mL (10%), 27.0µg/ml (23%), in which the former are
satisfactory to USP requirements for release ≤ 10% at pH 1.2. For the systems at pH 7.4, it was possible to
verify a control of the drug release, at 6 hours, SD10%, SD20%, SD30%, release 5%, 29% and 31%,
respectively, coming upon the slower generation of delayed supersaturation, which retard nucleation events, in
addition to having na increased solubility in SD20% and SD30% up to 200%.The association of polymers had the
ability to turn into the amorphous form all systems studied, besides increasing the solubility and slowly release
the drug pH dependent in pH 7.4. Therefore, relevant results were obtained to execution of future studies at this
issue.
Financial Support
CAPES and CNPq
295
PREPARATION AND CHARACTERIZATION OF GASTRORETENTIVE FLOATING
SYSTEM OBTAINED BY BINARY SOLID DISPERSION
Nathalia Cristina Gonçalves De Rosa
College of Engineering of Bioprocess and Biotechnology of the University of Sorocaba, São Paulo, Brazil
Vanessa Romero da Silva
College of Engineering of Bioprocess and Biotechnology of the University of Sorocaba, São Paulo, Brazil;
Biomaterials and Nanotechnology Laboratory University of Sorocaba, Sorocaba, São Paulo, Brazil
Biomaterials and Nanotechnology Laboratory University of Sorocaba, Sorocaba, São Paulo, Brazil
Introduction: Gastric emptying is a physiological event that interferes with the drug bioavailability from
stomach and proximal duodenum. A gastroretentive floating system (GFS) can be defined as a system which is
retained in the stomach for sufficient time against all the physiological barriers and contribute to improve the
therapeutic activity and the bioavailability of drugs.To fulfill all these objectives, an optimized GFS should
possess physical properties like being smaller in size, high buoyancy, and encapsulation efficacy along with the
controlled release of drug in the gastric environment. Methodology: The technique of solvent diffusion and coprecipitation of Eudragit S100 (ES100) and polyvinyl alcohol (PVA) was used for the preparation of the GFS. The
process was developed in an isolated system with control of temperature (60ºC), pressure (1atm) and agitation
(300rpm). These are the experimental factors which were selected from the characterization of the particles:
morphology (shape), size (Dm), zeta potential (ZP), polydispersity (PI) and pH. The responses parameters were
monitored for 30 days to assess system stability. Results and Discussion: The spherical-shaped particles were
uniform and had smooth surface. At time zero, there were identified the ZP -12.99mV (± 1.04); Dm 255.7nm and
PI 0.271. During a 30-day storage time, the ZP reduced and remained stable in 20.84mV; the Dm decreased from
255 nm to 200nm and the PI remained unchanged between 0.13 and 0.2. The pH remained in the rage of 5 and 6.
Conclusion: The obtained results of the qualitative screening study were according to the binary solid dispersion
theory and, the results show that the binary solid dispersions prepared by co-precipitation are an alternative to
obtain floating gastroretentive particles.
Financial Support
FAPESP, FINEP
296
SOLUBILITY EVALUATION AND THERMAL CHARACTERIZATION OF DISPERSED
SOLID SYSTEMS WITH PVP / PEG AND NEW DERIVATIVE 2-AMINOTHIOPHENE
Nathália Ribeiro Calazans Brandão da Silva
Danielle Lima Bezerra de Menezes
Walter Ferreira da Silva Júnior
Cícero Flávio Soares Aragão
Francisco Jaime Bezerra Mendonça Júnior
Introduction: 6CN is a prototype derived 2-aminothiophene with low water solubility and high
crystallinity. Indication of anti-inflammatory activity was obtained by in vitro test previously conducted. The use of
solid dispersions (SD) has proved effective in increasing the solubility of poorly soluble substances. PVP K-30
and PEG 6000 are polymers with low toxicity and high solubility that are used to form SD. This study aimed to
evaluate the solubility and the thermal charactheristics of systems. Methods: Physical mixture (PM) and SD have
been developed by kneading (KN) and rotatory evaporator (EV) methods in 1: 1 w/w ration. These systems were
characterized by Differential Scanning Calorimetry (DSC), Termogravimetry (TG) and Derivative Termogravimetry
(DTG) and a phase solubility test was performed with the samples quantified by UV/Vis spectrophotometry.
Results and Discussion: The DSC data showed the thermal behavior of 6CN by two endothermic events: the first
observed between 138 - 153°C indicating the melting of the crystalline structure and the second between 188 253°C corresponding to degradation of the prototype, confirmed by TG as a single mass loss step in the
temperature range 165 - 252°C. In the PM and SD analysed in DSC was observed the disappearance of
characteristic endothermic events 6CN, indicating that the prototype can be solubilized in the polymer matrix. The
thermogravimetric curves of PMs and SD showed an increase in 6CN degradation temperature, indicating that the
polymers promoted a thermal protection to the prototype. The solubility studies showed non-linear relationship and
the polymers were not able to significantly alter the solubility of the 6CN. In vitro dissolution test will be performed
to check if there was an increase in the dissolution rate. Conclusion: Although polymers do not influence the
solubility of 6CN, thermal analysis showed that the formation of solid dispersions promoted increased thermal
protection to the prototype.
Financial Support
This study has financial support from CNPq.
297
VALIDATION OF ANALYTICAL METHODOLOGY AND CHARACTERIZATION OF
POLYMERIC NANOCAPSULES CONTAINING HESPERETIN
Bruno Santos Lima
Introduction: Hesperetin (HT) is a flavonoid found abundantly in citrus fruits and it has pharmacological
effects such as antioxidant, anti-inflammatory, antiplatelet and vasoprotective. However, in plants, HT is often
glycosylated (hesperidin), and therefore has low solubility. In this context, the incorporation of HT in polymeric
nanocapsules (NC), can improve your solubility, bioavailability and stability. Methods: The analytical method to
HT quantification was developed and validated according with parameters (linearity, limit of detection and
quantification, precision, accuracy and robustness) established by International Conference on Harmonization
guidelines. The NC-HT (1 mg/ml) were obtained by interfacial deposition pre-formed polymer method and
characterized by pH, zeta potential, polydispersity, particle diameter, stability and in vitro permeation. Results and
Discussion: All validation parameters showed excellent relative standard deviations (RSD) and R2 0.9991 proving
that the method is linear, sensitive, accurate and precise. The NC-HT dimensions, obtained by laser diffraction,
showed an average particle diameter of 277.33 ± 98.42 nm, diameter 50% of the particles D(0.5)=94.33±40.42
nm, span value of 1.39±0.16. In addition, the particles were analyzed by dynamic light scattering having a
diameter 96.19±4.51 nm, polydispersity index 0.098±0.030 and zeta potential -7.64±1.0. Additionally, in the trace
analysis of nanoparticles were observed 2.87x1012 particles/mL e D(0.5)=225±51 nm. The encapsulation
efficiency was 98.65 ± 0.80% and the formulations were stable for 14 days with pH 6.52 ± 0.27. With the in vitro
permeation study, it was observed that the NC-HT were able to penetrate and permeate into all skin layers, acting
as a good formulation for topical use, suitable for relaying the HT to the via systemic action. Conclusions: The
studied formulation, showed a high potential to the incorporation in a suitable pharmaceutical form for topical use.
Financial Support
CAPES, CNPq, and FAPITEC/SE
298
INFLUENCE OF THE SPRAY-DRYING PROCESS ON THE MUCOADHESIVE
PROPERTIES OF CARVEDILOL-LOADED NANOCAPSULES
Paula dos Santos Chaves
Programa de Pós-Graduação em Ciências Farmacêuticas, Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul, Porto
Alegre, RS, Brazil.
Alegre, RS, Brazil.
Departamento de Química Orgânica, Instituto de Química, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil.
Alegre, RS, Brazil.
Alegre, RS, Brazil.
Introduction: The administration of drugs by the sublingual route can be an interesting alternative to
improve the bioavailability of molecules with liver metabolism. The use of a mucoadhesive carrier may be a
strategy to avoid the drug removal by the salivary flux. In this study the influence of the spray-drying process on
the mucoadhesive properties of polymeric nanocapsules for sublingual administration of carvedilol were
investigated. Methods: Carvedilol-loaded nanocapsules containing Eudragit® RS100 (EUD-NC) or poly(ε
caprolactone) (PCL-NC) were produced by interfacial deposition of the preformed polymer. Their mean size and
polydispersity (laser diffraction-LD) and zeta potential (electrophoretic mobility) were measured. The suspensions
were spray-dried using 10% of polyvinilpyrolidone:lactose (1:1 g/g). The recovery of the original nanoparticles after
aqueous redispersion of powders was checked by LD and scanning electron microscopy. Mucoadhesive
properties of nanocapsules were evaluated by interaction with mucin (colorimetric method) or sublingual mucosa
(tensile stress). Results and discussion: The mean size of EUD-NC and PCL-NC were 149±1 and 191±27 nm,
respectively. Both formulations showed narrow particle size distribution. Zeta potential values were +7±3 and -8±1
mV, for EUD-NC and PCL-NC, respectively. Original nanoparticles were recovered by both powders.
Nanocapsules were able to interact with mucin and this ability was not influenced by the spray-drying process.
EUD-NC interacted with a higher quantity of mucin than PCL-NC. A higher work was necessary to detach the
interaction between sublingual mucosa and EUD-NC or the respective powder. Conclusion: Polymeric
nanocapsules showed promising mucoadhesive properties. EUD-NC showed a better adhesive property than
PCL-NC. The spray-drying process did not alter their adhesive properties and may be further studied for the
development of innovative sublingual dosage forms.
Financial Support
Fapergs/RS/Brazil, CNPq/Brazil and CAPES/Brazil.
299
Preparation and characterization of inclusion complexes between βcaryophyllene and
methylβcyclodextrin
Pauline Sousa dos Santos
Federal University of Piauí
Rui Albuquerque Carvalho
University of Coimbra, Portugal
Francisco José Baptista Veiga
Ana Rita Figueiras
Lívio César Cunha Nunes
Federal University of Piauí
βcaryophyllene (BCP) is a sesquirterpene that has attracted attention from pharmacologists because it
presents a variety of biological activities. The main goal of this study was to investigate the formation of inclusion
complexes between BCP and methylβcyclodextrin (MβCD) in aqueous solution and in solid state and to
characterize the performed complexes using different methods. Methods: Phase solubility studies were performed
using MβCD following the method described by Higuchi and Connors, 1965. Solid systems were prepared in an
equimolar ratio BCP:MβCD using: kneading (KN), rotatory evaporation (ROE) and lyophilization (Lph). The
obtained binary systems were characterized using Fourier transform-infrared spectroscopy (FTIR), differential
scanning calorimetry (DSC) and nuclear magnetic resonance spectroscopy (NMR). Results and Discussion:
Phase solubility studies showed that BCP solubility increased 9.85-fold when compared with the BCP intrinsic
aqueous solubility, in the presence of the maximum of MβCD (40 mM) and the ks value calculated was 218.76 M,
diagram type: AL, and stoichiometry: 1:1. In the FTIR spectrum the most intense peak for BCP observed was at
886 cm-1 and it was not observed when the spectra of the inclusion complexes were analyzed, probably due to a
restriction of the vibration related to the complexation process. In DSC curves the absence of volatilization peak
of BCP in the thermograms of the ROE and Lph systems also indicates the possible formation of inclusion
complexes. The calculated upfield shifts in 1H-NMR indicated that the methylated part of BCP is involved in the
complexation, however the calculated values indicated a weak interaction. Conclusions: The results presented in
this study prove that the BCP solubility was increased in the presence of MβCD, stable inclusion complexes can
be prepared at a 1:1 BCP:MβCD and the complexation occurs preferentially by the doubly methylated part of
BCP.
Financial Support
CAPES
300
DEVELOPMENT AND STABILITY STUDY OF A PEDIATRIC ORAL SOLUTION OF
BACLOFEN 2 mg/mL
Priscila Rosa
Juliana dos Santos
Introduction: Baclofen is a skeletal muscle relaxant effective in reducing spasticity and muscle spasm
in patients with spinal cord injuries. Studies in Brazilian hospitals revealed the lack of some drugs in liquid form
for oral administration, which are necessary for patients that require non-standard doses, such as pediatric
patients. Baclofen is marketed as tablets and was indicated as a drug required in liquid form, for oral use. Thus,
the purpose of this study was to develop an oral solution of baclofen suitable for pediatric administration.
Methods: The oral solution was composed by potassium sorbate, citric acid, sucrose and baclofen at 2 mg/mL.
To the stability study, the formulation (n=3) was stored in a climatic chamber at 25°C/75% UR and under
refrigeration, during 30 days. The appearance, pH, baclofen content and microbial limits of the formulation were
evaluated just after preparation (zero time) and after 30 days of storage. To assay the preparation, we used a LC
stability indicating method previously validated. The microbial limits were performed by the estimation of the
number of viable aerobic microorganism. Results and Discussion: At zero time, the formulation was clear and
colorless, presented pH of 4.45 ± 0.02 and content of 100.21 ± 2.00%. After 30 days of storage, no changes in
the appearance of the solution were observed, as well as the pH did not differed statistically (p>0.05) from the
initial value. The baclofen content were 99.71% and 98.29%, stored at 25 °C/75% UR and refrigerator,
respectively. The microbiological assay indicated that the solution remained within acceptable microbial limits.
Conclusion: A simple and new formulation of baclofen 2.0 mg/mL was proposed. The solution remained stable for
30 days in both studied conditions, and represents an alternative to the administration of baclofen by oral route.
Financial Support: CAPES, FIPE-UFSM.
Financial Support
CAPES, FIPE-UFSM
301
Nanoencapsulated imiquimod action on cervical cancer cell line (SiHa)
Paola Mello
Flávia Pires Peña
Andreia Buffon
Silvia Staniscuaski Guterres
Introduction: Cervical cancer is related to human papillomavirus infection (HPV), the drug used in
treatment is imiquimod, which causes adverse effects. Nanoencapsulation strategy aims to decrease adverse
effects related to the drug. This work aims to evaluate cytotoxic action of the new formulation during a
determinate time interval and acute and chronic effects on cervical cancer cell line. Methods: Two polymeric
nanocapsules suspensions were developed by the method of nanoprecipitation and named as: NCimiq and NC
(nanocapsules without imiquimod). The free drug dissolved in dimethyl sulfoxide (IMIQfree) was also evaluated for
comparison with the developed nanosystem. The formulations were tested in cell culture (SiHa), plated 4x103
cells per well for 24 hours to achieve semi-confluence. The results were assessed by flow cytometry technique
(BD Biosciences, San Jose, CA, USA) after treatment in 24, 48 and 72 hours. Results and discussion: After 24
hours it is noted that NCimiq decreased 60% of cell viability when compared to control and decreased cell
viability five times more than IMIQfree. NC also decreased cell viability (about 37%), probably due to copaíba oil
component, which has anti-inflammatory action. After 48 hours, it is noted that NCimiq decreased 87% of cell
viability when compared to control, whereas IMIQfree only 22%. Finally, after 72 hours of treatment with NCimiq
there were no more viable cells in the well, whereas IMIQfree decreased only 44% of cell viability. Therefore, it
can be observed acute (after 24 hours) and chronic effects (after 72 hours) of the nanoencapsulated drug on
cervical cancer cells and it is concluded that nanoencapsulated drug can decrease more cell viability than free
drug. Conclusions: The proposed formulation has potential to be used in treatment of cervical cancer since it was
more effective than the free drug.
Financial Support
CAPES, CNPq
302
DEVELOPMENT, CHARACTERIZATION AND EVALUATION OF THE ANTIFUNGICAL
ACTIVITY OF PLGA AND AMPHOTERICIN B NANOFIBERS COMPARED TO DIFFERENT
SPECIES OF ATCC CANDIDS AND CLINICAL STRAINS
Ramon de Oliveira Souza
Postgraduate Program in Biotechnology, Universidade Federal de São João del-Rei, Divinópolis, MG, Brazil
Marcelo Gonzaga de Freitas Araújo
Universidade Federal de São João del-Rei, Divinópolis, MG, Brazil
Tadeu Henrique de Lima
Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil
Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil
Juliana Teixeira de Magalhães
Postgraduate Pharmaceutical Sciences, Universidade Federal de São João del-Rei, Divinópolis, MG, Brazil
Gisele Rodrigues da Silva
Postgraduate Program in Biotechnology, Universidade Federal de São João del-Rei, Divinópolis, MG, Brazil
Introduction: The Amphotericin B (APB) is an antifungical drug which has several collateral effects to
the patient, due to its high toxicity. As an alternative, the polymeric nanofibers has been evaluated as interesting
drug delivery systems, considering the possibility of active principle vectorization, reducing the collateral effects
linked to the parenteral administration of APB. Methods: Nanofibers based on PLGA e APB were produced by
electrospinning technique. The characterization was accomplished by Fourier transform infrared spectroscopy
(FTIR) and Scanning electron microscopy (SEM). It was determined the uniformity of APB content in nanofibers
by spectroscopy at ultraviolet region, besides its profile of in vitro of liberation. The method of Agar diffusion
evaluates the antifungical action of these systems according to Candida albicans ATCC 26790, Candida glabrata
ATCC 2001, Candida krusei ATCC 34135, Candida tropicalis ATCC 28707 cultures, and also five clinical strains
of patients with vulvovaginal candidiases. Results and Discussion: The results obtained by FTIR showed that the
drug and polymeric nanofibers did not chemically interact. Eletromicrography using SEM made possible to identify
nanofibers production characterized by nanometric polymer fibers network. The uniformity test of content showed
homogeneity on a distribution of drug on the fibers. The kinectics of the in vitro release revealed the drug liberation
over time for a period of approximately 192 hours. Antifungal activity was verified by the formation of inhibition
zone to all the candida species present in the study. Conclusion: Thereby, these systems containing APB has
potentiality in biomedical applications, which allows the minimization of side effects to the patient, and can reduce
the need for frequent administration of a drug, due to its sustained and controlled liberation.
Financial Support
The authors wish to thank the UFSJ (Brazil), CNPq/MCT (Brazil) and Fapemig (Brazil) for the financial support.
303
PHYSICOCHEMICAL CHARACTERIZATION OF BETA-SITOSTEROL LOADEDPOLYMERIC NANOCAPSULES
Raquel Lautenchleger
Tatiele Katzer
Cristiane De Bona Da Silva
Ruy Beck
Introduction: Androgenetic alopecia is the most common cause of progressive hair loss. Testosterone is
converted to dihydrotestosterone (DHT) as a result of 5αreductase (5αR) activity. DHT activity and a chronic
perifollicular inflammation are responsible by the miniaturization of the hair follicles. This work presents a partial
characterization of polymeric nanocapsules containing βsitosterol (NCPCLBS), an anti-inflammatory substance
able to inhibit the activity of 5αR. Methods: NC-PCL-BS formulations were prepared (n=3) by the precipitation of
pre-formed polymer method and characterized after preparation (T0) and during 30 days (T30). The particle size
distribution was determined by laser light diffraction. For a more specific of particle size, polydispersity index
(PDI) and zeta potential the formulations were analyzed by photon correlation spectroscopy. High performance
liquid chromatography was used to evaluate the drug content and encapsulation efficiency (ultrafiltrationcentrifugation method) of the NC-PCL-BS. The pH of the formulations was analyzed by a calibrated pH meter.
Results and discussion: NC-PCL-BS showed particles exclusively within nanometric range, with mean size of
231 ± 34 nm and 211 ± 8 nm at T0 and T30, respectively, with a narrow particle size distribution (PDI at T0 = 0.18
± 0.08 and T30 = 0.14 ± 0.01). The βsitosterol content of NCPCLBS was 0.97 ± 0.03 mg/ml at T0 and 0.99 ±
0.03 mg/ml at T30. The entrapment efficiency was close to 100%.The pH decreased from 6.09 ± 0.17 to 5.27 ±
1.11 as a function of time. The zeta potential was negative at T0 (-9.1 ± 1.2 mV) and remained unchanged at T30
(-9.7 ± 2.3 mV). Conclusions: NC-PCL-BS presented suitable physicochemical characteristics during 30 days.
Considering the biodegradability and biocompatibility of the components to obtain the NC-PCL-BS, this
formulation will be further characterized and its potential on androgenetic alopecia treatment evaluated.
Financial Support
CAPES
Acknowledgments
The authors thanks CAPES for the financial support.
304
PRODUCTION, CHARACTERIZATION AND ASSESSMENT OF THERMAL
HYPERALGESIA OF CHITOSAN HYDROGELS COMPOSED BY POLYMERIC
NANOCAPSULES LOADING CAPSAICINOIDS
Ricardo Lorenzoni
Cleverton Kleiton Freitas de Lima
Ana Luísa Palhares de Miranda
Teresa Dalla Costa
Introduction: The peripheral neuropathic pain (PNP) is a complex disease, its treatments using creams
containing capsaicinoids are low effective and cause several side effects. Polymeric nanocapsules (NC) have
been used to improve therapeutic efficacy and reduce the side effects of drugs. Thus, we produced, characterized
and assessed in animal model of PNP, chitosan hydrogel containing NC loading capsaicinoids (CH-NCCaps),
comparing with commercial cream of reference on pharmaceutical market. Methods: NC suspensions were
prepared by interfacial deposition of preformed polymer. The characterization were realized by pH determination,
average particle size, polydispersity index (PDI), zeta potential (ζ) and drug encapsulation efficiency. The CHNCCaps were produced with manual mixing, and were characterized in terms of pH, presence of nanoparticles,
viscosity, and capsaicinoids content. The test which evaluates thermal hyperalgesia was performed in animal
model of PNP (approved by CEUA; UFRJ; number FARMÁCIA 04). Results and Discussion: The NC showed
pH acid; with average dimensions near 140 nm; low PDI, indicating unimodal size distribution; positive ζ; and
encapsulation efficiency near 100%. The CH-NCCaps showed pH acid; consistency of 20.38 Pa.sn; flow index of
0.70; and pseudoplastic behavior, which is suitable for cutaneous applications. The capsaicinoids content in CHNCCaps and in cream were 1.15 and 1.17 mg/g, respectively. The CH-NCCaps was more effective in reduction
of thermal hyperalgesia 24 hours after the first dose, effect that did not happen with the commercial cream.
Conclusions: The NC containing capsaicinoids showed nanometric size, small PDI and the drugs were highly
associated the NC. There was maintenance of nanometric structures in CH-NCCaps, which showed suitable
characteristics for topical application. CH-NCCaps was more effective against thermal hyperalgesia when
compared to commercial cream.
Financial Support
CNPq, CAPES and PPFCF.
Acknowledgments
CNPq, CAPES and PPFCF.
Ethical approval
approved by CEUA; UFRJ; number FARMÁCIA 04
305
DEVELOPMENT OF DHC LOADED-NANOSTRUCTURED LIPID CARRIER WITH
POTENTIAL ANTITUMOR ACTIVITY
Ruth Meri Lucinda da Silva
Thamiris Yumi Inoue
Tania Mari Belle Bresolin
Angela Malheiros
Introduction: Nanostructured lipid carriers (NLC) are composed of solid and liquid lipids, surfactants and
vehicle. These systems showed biocompatibility and improvement stratum corneum nanoparticles retention and
drug penetration. 2',6'-dihydroxy-4'-methoxydihydrochalcone (DHC) is isolated from Piper aduncum leaves, which
showed antitumoral and antimicrobial activities. In order to increase DHC cytotoxicity, DHC-loaded NLC was
developed. Methods: NLCs were prepared using microemulsion method and Glyceryl Behenate (GB) as lipid. To
DHC-loaded NLC optimization, three different variables were analyzed: surfactants (Polissobate - PO and
Ethoxylated Castor oil - ECO) and liquid lipid (Caprylic Capric Triglycerides - CCT or Isopropyl Myristate - IPM)
used and DHC:lipid ratio (1:20 and 1:40). NLC without DHC was prepared also. NLC properties of distribution
size, polydispersity index, morphology, zeta potential and encapsulation efficiency (EE) were determined. In vitro
release profile was evaluated using modified dissolution apparatus and hydroalcoholic solution 30% (v/v) as
release medium. Results and discussion: ECO was chosen as surfactant because NLC containing ECO had
better physical aspect, translucent colour during 14 days, higher zeta potential and PI between 0.3 and 0.4. NLC
containing CCT and Isopropyl Myristate IPM showed satisfactory results to particle size, PI and zeta potential.
Nonetheless, NLC containing IPM showed better PI than NLC containing CCT. Blank NLC size was about 15.75
to 23.35 nm over 30 days and of the DHC-NLC was about16.58 to 17.88 nm. The DHC encapsulation efficiency
was >99%. The release profile showed initially a quick release or burst effect, which can be related to DHC an
outer layer or NLC surface, then after 30 min, a slower release was observed, with 36% of DHC released after 24
h. Conclusion: DHC-NLC were successfully obtained with PI inferior to 0.4, no visual agglomeration, translucent
colour and stable formulation.
Financial Support
PRONEM/FAPESC/CNPq, UNIVALI.
306
OSCILLATORY RHEOMETRY OF POLYMER BLENDS COMPRISING POLOXAMER 407
AND CARBOPOL 974P®
Jéssica Bassi da Silva
The development of blends containing thermoresponsive and bioadhesive polymers can provide new
drug delivery systems. Thus, the present work describes oscillatory rheometry of bioadhesive thermoresponsive
systems containing poloxamer 407 (P407) and Carbopol 974P® (C974P), to be used as platform of
pharmaceutical systems. Ten polymer blends were prepared containing P407 (15 or 20%, w/w) and C974P (0.10,
0.15, 0.20, 0.25 or 0.50%, w/w). Oscillatory rheometry was performed using a MARS II (Haake®) controlled
stress rheometer, in oscillatory mode, at 5, 25 and 37 ± 0.1 °C. After the determination of the linear viscoelastic
region, frequency sweep analysis was evaluated from 0.1 to 10.0 Hz. The storage modulus (G'), loss modulus
(G"), dynamic viscosity (η') and the loss tangent (tan δ) were calculated by RheoWin 4.10.0000 (Haake®)
software. In each case, the oscillatory rheological properties of at least five replicates were determined. All
formulations showed viscoelastic properties that were oscillatory frequency, temperature and
polymer
concentration dependent. Thereby, the increase of oscillatory frequency was accompanied by G' and G" increase
for all systems, except G" of polymer blends containing 20% (w/w) P407 evaluated at 37 °C. Moreover, the
increase of oscillatory frequency decreased η' and tan δ for most systems, whereas, tan δ of polymer blends
containing 20% (w/w) P407 at 37°C were not influenced by oscillatory frequency. Furthermore, the increase of
temperature and polymer content increased G', G", η', while tan δ was decreased. Importantly, the formulations
15/0.50, 20/0.25, 20/0.50 at 5 °C, 15/0.50, 20/0.10, 20/0.15, 20/0.20, 20/0.25 and 20/0.50 at 25 °C and all
formulations evaluated at 37 °C exhibited G' higher than G", so are classified as viscoelastic. Polymer blends
with viscoelastic behavior showed rheological properties suitable to be used as pharmaceutical systems.
Financial Support
CAPES, CNPq, Fundação araucária, FINEP.
307
In vitro release test of clozapine from nanocapsules prepared with different surface
characteristics
Sandra Elisa Haas
College of Pharmacy, PostGraduate Program in Pharmaceutical Sciences, Federal University of Pampa, Uruguaiana, RS, Brazil.
Eduarda Martini
College of Pharmacy, Federal University of Pampa, Uruguaiana, RS, Brazil.
Renata Bem
College of Pharmacy, Federal University of Pampa, Uruguaiana, RS, Brazil.
Introduction. The clozapine (CZP) is an atypical antipsychotic which has some serious side effects
such as hepatotoxicity, cardiotoxicity and agranulocytosis. Then, its clinical use is reduced to cases of refractory
schizophrenia. Furthermore, CZP undergoes extensive presystemic metabolism, reducing the systemic
bioavailability. In this context, we prepared CZP loaded-nanocapsules (NC) with different coatings aiming improve
the absorption. In this work, Polysorbate 80 (P80), PEG or chitosan (CS) were used as coating and the ability of
these coatings on modulate the clozapine release of in vitro simulated gastrointestinal medium were evaluated.
Methods. NC were prepared by nanoprecipitation method and characterized in terms of particle size distribution,
zeta potential, pH, drug loading and encapsulation rate. The in vitro release profiles were evaluated by dialysis
bag technique. Briefly, the NCs or CZP solution were added inside the bag and in contact with the media pH 1.2
during 120 min. After that, the bag was placed inside the 6.8 pH solution for 180 min approximately. Samples
were collected in pre-determinate times and the drug was evaluated at Shimadzu spectrophotometer. Results and
discussion. The results showed that the NC presented particle size between 2000 and 238 nm. The zeta potential
values were negative for P80 and PEG-coated NC and positive when CS was used. pH values were acid from
CS-formulations and unloaded-NC. CZP-loaded P80 and PEG-coated NC showed pH=7. CZP were completely
released in pH acid while NC had slower release between 54 and 61% in this pH and ranged from 62 to 72% after
3 h in pH 6.8. Conclusions. The nanoparticulate systems showed a more controlled release and we intend to
evaluate the in vivo absorption process of CZP from coated-NC.
Financial Support
FAPERGS; CNPq, UNIPAMPA
Acknowledgments
UNIPAMPA
308
IN VITRO EVALUATION OF PHOTOPROTECTIVE EFFICACY OF COMMERCIAL
SUNSCREENS BY BIOLOGICAL PARAMETERS
Sônia Aparecida Figueiredo
University of Sao Paulo
Dayane Cristina de Moraes
Ana Luiza Scarano Aguillera Forte
Fernanda Maria Pinto Vilela
Juiz de Fora Federal University
Maria José Vieira Fonseca
Introduction: The use of sunscreens increases worldwide. Therefore, there is concern with the
sunscreens effectiveness and safety. These factors are usually determined by measure: i) protection factor (SPF,
UVA-PF) and ii) skin pigmentation (IPD, PPD) in human volunteers. However, these measure do not reflect the
damaging effects in cellular structures and components induced by ultraviolet radiation (UV), such as, DNA, lipids
and proteins. In this context, the major goal was analyze different cell lines and biological parameters, intending to
stablish an accurate and precise in vitro method to which assesses the photoprotective efficacy of sunscreen and
guarantees product safety. Methods: Firstly, different skin cell cultures (L929, HaCaT and MRC5) were exposed
to several UVA and UVB radiation doses and biological parameters were assessed to set the best radiationresponse model. Next, the influence of UV radiation on cell viability, formation of lipid peroxides and production of
reactive oxygen species (ROS) were assessed. The samples of commercial sunscreens were applied in a quartz
plate placed on top a microplate filled by cell cultures. Results: Among the cell lines and biological parameters
tested, the cell viability and lipid peroxidation measured in L929 cells were the most promising to evaluate the
efficacy of sunscreens, as well as, allowed to discriminate the photoprotective potential from different SPFs
formulations against UVB radiation. The ROS generation, expressed in HaCaT lineage, proved to be a promising
biological test to discriminate the photoprotective potential from sunscreens with different SPF or SPF/PPDs
exposed to UVA radiation. Conclusion: Thus, after confirming the accuracy and precison of these methods, it
could be suggested that the results open up high expectatives for in vitro assessments of sunscreen efficacy
based on different and complementary parameters from those evaluated in vivo.
Financial Support
CNPq, FAPESP and CAPES
Acknowledgments
CNPq for the scholarship granted
309
CONTROL OF QUALITY AND PERFORMANCE TESTS FOR EVALUATION OF
INHALATION FORMULATION CONTAINING SILDENAFIL
Taízia Dutra Silva
Universidade Federal dos Vales do Jequitinhonha e Mucuri (UFVJM)
Ana Carolina Guimarães Ribeiro
Jackson Antônio Lamounier Camargos Resende
Universidade Federal Fluminense (UFF)
Raquel Braga Dutra Gonçalves
Universidade Iguaçu (UNIG)
Cristina Duarte Vianna-Soares
Introduction. Pulmonary arterial hypertension (PAH) is a severe disease responsible for a high mortality
in the affected population. The treatment has limitations due to severe adverse effects, high cost and limited
access, especially for iloprost and bosentan drugs. Hence, we intend to develop a new therapeutic option using
sildenafil citrate (SILC), in the dry powder form for inhalation. The pulmonary administration is an excellent
alternative for the treatment of severe lung diseases providing maximized local effect, little or no systemic
absorption and reduction of adverse effects. Methods. In order to reach the target organ, the drug particle size
was reduced a few micra by milling. The SILC of reduced particle size was mixed with appropriate diluent and
dispensed in capsules. The formulation was characterized by techniques: powder X-ray diffraction, laser
diffraction and scanning electron microscopy, focusing on the distribution and size of a proper particle inhalation
(1-5 µm). To ensure the quality and efficacy of inhaled formulation, performance tests such as delivered-dose
uniformity (DDU) and the aerodynamic size distribution (ASD) were accomplished according to ANVISA or official
compendia requirements for inhalation dosage forms. Results and discussion. The results of the particles
micronization showed a size 2.93 µm and 9.08 µm for the percentiles P50 and P90%, respectively. For DDU, a
range of 92.42-109.05% of dose released was obtained from the inhaler device, results in agreement with the
range 75.0-125.0%, specified by the US Pharmacopeia. For ASD, a fraction of 35% of fine particles (<5 µm, FFP)
was satisfactorily obtained. Conclusions. The developed formulation, in the dry powder form, achieved the
appropriate characteristics for pulmonary administration with suitable FFP and aerodynamic size for SILC.
Additionally, it can be an excellent alternative for the treatment of PAH.
Financial Support
FAPEMIG, Brazil.
Acknowledgments
The authors are grateful to FAPEMIG, CNPq (Brazil) for the research financial support and Instituto Vita Nova (Hortolândia, S P, Brazil) for the kind donation of
sildenafil citrate raw materials.
310
STABILITY STUDY OF EFF-Cg-LOADED PLGA NANOPARTICLE SUSPENSIONS AT
AMBIENT TEMPERATURE
Talitha Caldas dos Santos
Mariana Alves Batistti
Caroline Ortmann
Flavio Henrique Reginato
Claudia Maria Oliveira Simões
Angela Machado de Campos
Introduction: The development of novel drug delivery system for herbal extracts has remarkable
advantages over conventional phytopharmaceuticals such as enhancement of bioavailability and pharmacological
activity. Furthermore, the long term storage stability of the nanoparticles is an important parameter to development
a formulation. Thus, one major requirement is the development of formulations, which can be reproducibly
prepared and stored for an extended period of time with full maintaining of particle integrity and biological activity.
The aim of this study was to prepare PLGA nanoparticles containing enriched flavonoid fraction of Cecropia
glaziovii Snethl (EFF-Cg) and to assess the stability for a period of 60 days. Methods: Six nanoparticles
formulations prepared with different stabilizer type of the organic phase and concentration of poloxamer 188 in the
internal aqueous phase were stored under static conditions at ambient temperature. The physical stability was
monitored by visual inspection of appearance of particles agglomerates, sediments and phase separation. Also,
the nanoparticles were evaluated for particle size, polydispersity index (PDI) and zeta potential by dynamic light
scattering, and the free chemical marker in the aqueous external phase was quantified by HPLC. Results and
Discussion: Visually was not detected phase separation phenomenon. After 60 days of storage, all formulations
presented particles sedimentation and the size and PDI values indicated the presence of the two size
populations. The zeta potential values of formulations were greater than -30 mV during all study. It was observed
an increase of free chemical marker on the external phase. However, all changes were less pronounced in the
formulation prepared with sorbitan monooleate. Conclusions: The more stable formulation in the assessed
conditions was tht it contained sorbitan monooleate and the 0.5% (w/v) of poloxamer 188 added in the organic and
aqueous phases, respectively.
Financial Support
CAPES
311
Development and evaluation of the physiomechanical properties of
thermoresponsives hydrogels for vaginal administration.
Márcia Araújo Rebelo
Vaginal retention, release time and the absence of local irritation are the main challenges for the
development of pharmaceutical forms for
local
administration.
Thermoresponsives
systems
with
physiomechanical properties appropriate and prepared with biocompatible material may be useful to increase the
retention time, control the drug release, reducing the local irritation and improve adherence to treatment. The
physiomechanical properties and the release profile of thermoresponsives hydrogels composed of poloxamer 407
(P407), hydroxypropyl methylcellulose K4M (HPMC K4M), and chitosan (Ch) has been developed for vaginal
application and evaluated in vitro. Curcumin (CUR), derived from Curcuma longa, was used as model drug. The
hydrogel was prepared by cold dispersion of the polymers in buffer citrate-phosphate pH 4.5. The CUR's load in
hydrogels it was 400ug.ml-1 in free form or in solid dispersions (SD) with P407 (1:2). The dissolution rate of CUR
and CUR's stability in the SD was evaluated. The physiomechanical properties of the hydrogel (pH, viscosityTsol-gel, hardness, compressibility, adhesiveness, cohesiveness, mucoadhesion and erosion rate) were
evaluated. The best results were obtained for SD CUR:P407 (1:2) obtained by co-precipitation. The results for
pH, viscosity and erosion rate were respectively 4.05±0.01, 8,100cP and 51.3% (90min). The results for the
physiomechanical properties of the hydrogel for hardness, compressibility, adhesiveness, cohesiveness and
mucoadhesion were respectively 24.1±0.45g, 286.21±10.64g.s-1, 30.00±1.22g.s-1, 20.81±1.65g e 8.53±0.41g. The
solid dispersion (1:2) with P407 increased dissolution rate and chemical stability of CUR. The dissolution profile
of CUR contained in the hydrogel showed a controlled release. The thermoresponsives hydrogels obtained with
P407, HPMC K4M and Ch have adequate physiomechanical properties for vaginal administration with modified
release for CUR.
Acknowledgments
UNISO, FAPESP, CNPQ
312
EVALUATE OF DRUG-DRUG INTERACTION BETWEEN APIs RECOMMENDED IN
ANTIRETROVIRAL PAEDIATRIC THERAPY
Thaisa Cardoso de Oliveira
Isabella Gonçalves Matos
José Lamartine Soares Sobrinho
Mônica Felts de La Roca Soares
The aim of this study was to evaluate the thermal behavior of zidovudine (AZT), nevirapine (NVP) and
lamivudine (3TC) in order to predict possible physical-chemical interactions among these APIs. Analysis with
DSC and TG/DTG were performed with APIs alone besides binary physical mixtures in the proportions 1:1 (M/M)
(AZT:3TC, AZT:NVP, NVP:3TC) and ternary physical mixture 1:1:1 (M/M/M) (AZT:3TC:NVP). DSC curve shows
the endothermic melting of AZT at 120°C, TG/DTG curves shows decomposition events at 182°C, respectively.
DSC curves of 3TC and NVP shows melting in 173 and 242°C, respectively and TG/DTG curves of them shows
events of decomposition at 205 and 202°C. DSC curve of AZT:3TC showed a single endothermic peak with
Tonset 108,9°C, but 3TC melting range became extended leading to a suspicion of an eutectic mixture formation.
For AZT:NVP the AZT melted at 108°C, however it wasn't possible to detect NVP endothermic melting peak
because it is in the same range of AZT decomposition exothermic peak. According to TG/DTG curves, the order
of stability for the isolated drugs is 3TC> NVP>AZT and for the binary mixtures is NVP+3TC > AZT+3TC >
AZT+NVP. Thus, it was observed that 3TC had a major influence on the stability of the binary mixtures. In the
binary mixtures, it is possible to see that NVP is easily influenced by the others drugs, which is proved by the
weight loss that happened in next to 3TC and AZT, respectively. DSC curve of AZT:3TC:NVP showed an
endothermic event Tonset in 107°C which solubilization of 3TC after AZT fusion, besides that, it cannot verify any
melting peak referring to NVP. It was observed that TG/DTG curve of AZT:3TC:NVP contain thermal events of
each drug isolated with Tonset 197,3°C. In conclusion, it is supposed that the process of drug melting in binary
and ternary mixtures were influenced according to the components of the system.
Financial Support
FACEPE
Acknowledgments
The authors acknowledge FACEPE for financial support in this research. We also thank professor Adriano Antunes de Souza Araújo from Laboratório de
Ensaios Farmacêuticos e Toxicidade (LEFT) of the UFS, Aracajú-SE, Brazil.
313
DEVELOPMENT OF POLY NANOPARTICLES (LACTIC-CO-GLYCOLIC ACID) (PLGA)
ENCAPSULATION FOR PROTEINS: A STUDY WITH L-ASPARAGINASE
Thayse Silva Medeiros
Sonaly Lima Albino
Ísis Valeska Freire Lins
Maria Salete da Silva Rodrigues
Alexsandra Conceição Apolinário
Carlota de Oliveira Rangel Yagui
Adalberto Pessoa Júnior
Introduction: L-asparaginase (ASNase) is an antineoplastic agent used in the treatment of acute
lymphoblastic leukemia, which has disadvantages as immunogenicity and short half-life. Nanoparticles produced
by the double emulsification method consist of an interesting alternative to increase the circulation time of
ASNase and decreasing immunogenicity. One of the most used polymers in obtaining this pharmaceutical form is
poly (lactic-co-glycolic acid) (PLGA), due it is biocompatible and biodegradable. In this study, we investigated the
encapsulation ASNase in PLGA nanoparticles. Methods: Initially it was prepared an aqueous solution of the
enzyme at 10 mg / ml. For the first emulsion, it was weighed 0.05 g of PLGA molecular weight MW = 24-38 kDa,
and added 2.5 mL of chloroform plus 250 uL of enzyme solution. The resulting system was subjected to
sonication for three cycles of 60 seconds at 50% power. To obtain the second emulsion, the system was poured
into 10 mL of aqueous solution of polyvinyl alcohol (PVA) at 1% and subjected to sonication for three cycles of 60
seconds at 50% power. The system was then submitted to evaporation of the solvent under reduced pressure for
24 hours. The obtained nanoparticles (100-fold diluted samples) were analyzed by dynamic light scattering on
Nanotrac wave device (MN Model 401). Results and Discussion: average size of nanoparticles were obtained
from 363nm and polydispersity index (PDI) 0.4. Conclusion: Based on these preliminary results, we can conclude
that the double emulsification method was effective in producing PLGA nanoparticles in the presence of ASNase
enzyme. However, it is necessary to quantify the ASNase encapsulated in the system so that one can then
evaluate the therapeutic efficacy of the formulation.
Financial Support
UEPB, FAFESP e CNPq
314
TRANSDERMAL DELIVERY OF ISONIAZID AS AN APPROACH TO OVERCOME
HEPATOTOXICITY EVENTS
Thiago Caon
Federal University of Santa Catarina
Cláudia Maria Oliveira Simões
Marcos Antônio Segatto Silva
INTRODUCTION The isoniazid (INH) is highly effective in anti-tuberculosis therapy, but its oral
administration may lead to poor patient compliance due to undesirable side effects such as hepatotoxicity events.
In view of the fact that the transdermal administration of drugs is well-known in order to overcome hepatic first
pass effects, this route was exploited for delivering INH. Furthermore, cases of cutaneous tuberculosis have been
recently reported, expanding the research interests in this field. METHODS INH was applied to porcine ear skin in
Franz diffusion chambers alone and with 5% (w/V) of three chemical penetration enhancers (menthol, limonene or
Transcutol®). Thermal and spectroscopic studies were performed to elucidate the mechanism of action of
chemical enhancers and drug-related transport type. RESULTS AND DISCUSSION The permeation rate of INH
was sufficient to ensure a systemic therapeutic effect, which have been shown by combining mathematical and
experimental models. The high hydrophilicity of the drug explains its lower partitioning into the skin compared to
intestinal mucosa. A follicular transport is preferably used for INH, which require certain action of excipients on
this transport route or extraction of lipids from skin. Menthol did not affect the permeation parameters of drug and
increased INH retention in skin (1.4-fold). Transcutol® reduced significantly the cutaneous permeation of INH,
probably due to a depot effect and an increase in degree of order in lipid and protein domains. Limonene increased
the diffusivity (2.8-fold) and permeation flux (1.5-fold) of INH in skin, which may be associated to higher degree of
disorder in lipid bilayers as suggested by DSC and FTIR analyses. CONCLUSION INH can be efficiently
delivered by skin route and topical or systemic effects may be achieved depending the selected excipients. A
positive correlation between log P of chemical enhancers and permeation rate was observed.
Financial Support
FAPESC/CNPq/CAPES
Acknowledgments
We would like to acknowledge the Brazilian funding agencies FAPESC, CNPq/MCTI and CAPES/MEC (PNPD fellowship¿001/2010) for fi nancial support
Ethical approval
Not applicable
315
EVALUATION OF THE INCLUSION COMPLEXES OF ISOPULEGOL WITH α AND ß
CYCLODEXTRINS: STRUCTURAL CHARACTERIZATION AND MOLECULAR DOCKING
Pharmacy Department, Federal University of Sergipe
Igor Araújo dos Santos Trindade
Physiology Department, Federal University of Sergipe
Isopulegol (ISOP) is a monoterpenoid alcohol which is present in the essential oils of several
therapeutic plants. The aim of this work was to increase the solubility of ISOP by formation of inclusion
complexes with α and βcyclodextrins (α and βCD) through three different methods: physical mixture (PM),
paste method (PC) and slurry complexation (SC). Complexes were prepared and characterized using differential
scanning calorimetry (DSC), thermogravimetric analysis (TG/DTG), Karl Fischer titration (KFT) and scanning
electron microscope (SEM). The amount of ISOP entrapped in the inclusion complexes was determined by gas
chromatography-mass spectrometry (GC/MS). Guest-host interactions have been investigated using nuclear
magnetic resonance and molecular docking. Thermal analysis results of the α or βCD/ISOP revealed the
formation of a complex mainly through the PC and SC methods for α and βCD, respectively, because the
second step of mass loss in the TG/DTG (114-312 °C) exhibited mass losses of 3.0 and 5.7%, respectively. KFT
results suggested that the formation of the inclusion complexes improves the thermal stability of ISOP molecule.
SEM images showed decreased size of the crystals, especially in the SC products. The GC/MS analyses
demonstrated that the PC method was the best to form complexation with αCD (48.8%); for βCD, the SC
method exhibited strong complexation (68.3%). Furthermore, 1D and 2D 1H NMR revealed chemical shift
changes and intermolecular cross-peaks observed between protons of ISOP and cyclodextrins suggesting
inclusion process by non-covalent interactions, except for physical mixture utilizing αCD. The docking study
demonstrated that αCD/ISOP inclusion complex was found to be highly probable and energetically favorable
model than the βCD/ISOP inclusion complex.
Financial Support
316
PRELIMINARY CHARACTERIZATION OF CARRAGEENAN AND XATHAN GUM
SEMISOLID FORMULATIONS CONTAINING KETOPROFEN-LOADED POMEGRANATE
SEED OIL NANOEMULSIONS
Verônica Ferrari Cervi
Luana Mota Ferreira
Letícia Cruz
INTRODUCTION Nanoemulsions (NE) are stable systems with characteristics such as small size,
drug protection against degradation and controlled release. In previous works, pomegranate seed oil NE
containing ketoprofen (KP) were prepared. However, intending a topical administration, it is necessary to develop
a semisolid formulation. Thus, polysaccharides like carrageenan (GC) and xanthan (GX) gum have interesting
properties such low toxicity, gel formation and biocompatibility. So, the aim of this work was the KP
nanoemulsions incorporation in a semisolid preparation employing these gums. METHODS NEs were prepared by
spontaneous emulsification method. After this, GC and GX were employed to develop a semisolid formulation
(SF), at 3% concentration, with a 2:1 ratio, respectively (n=3). Characterization were performed in terms of mean
droplet size and polydispersity index (PDI), measured by photon correlation spectroscopy, using a ZetaSizer
(Zetasizer Nanoseries, Malvern Instruments). The pH values were determined by directly immersion of an
electrode potentiometer and the spreadability test was assessed using an apparatus plate. Moreover, High
Performance Liquid Chromatography (HPLC) was used to determine the total KP content in NEs and SF.
RESULTS AND DISCUSSION NEs droplets diameter was below 250 nm, PDI was less than 0.2 and pH values
were in the acid range. Furthermore, SF mean droplet size were 257 nm, which shows that NE droplet size was
not altered. Therefore, PDI was 0.35 and pH values were in the neutral range. Spreadability results showed a
mean factor of 3.19 mm²/g, considered suitable for topical formulations. Thus, HPLC analyses showed a mean
KP total content around 90% in SF and 95% for the NE, according to KP theoretical amount (1 mg/g).
CONCLUSION As a conclusion, this preliminary characterization study showed that GC and GX could be a
potential material for semisolid formulation development and further nanostructured systems incorporation.
Financial Support
CNPq
317
OCULAR BIOCOMPATIBILITY OF NANOFIBERS COMPOSED ON
POLY(CAPROLACTONE) AND DEXAMETHASONE ACETATE
Wellesin Antonio Carvalho Araújo
Universidade Federal de São João Del Rei (UFSJ)
Welleson Antonio Carvalho Araujo
Universidade Federal de São João Del Rei (UFSJ)
Tadeu Henrique Lima
Gisele Rodrigues da Silva
Francine Behar-Cohen
Université René Descartes Sorbonne Paris Cité, Paris 75006, France, Assistance Publique Hôpitaux de Paris, Hôtel-Dieu de Paris, Paris
75004, France
Min Zhao
Université René Descartes Sorbonne Paris Cité, Paris, INSERM, U872, Team 17, Centre de Recherche des Cordeliers, Paris 75006,
France
Armando Silva-Cunha
Introduction: Biocompatibility is a requirement for the development of nanofibers for ophthalmic
applications. In this study, nanofibers composed on poly(e-caprolactone) and dexamethasone acetate (DX-PCL
nanofibers) were synthesized using the electrospinning technique. The ocular biocompatibility of the polymeric
nanofibers, the by-products of the polymers and the dexamethasone released from the nanofibers was
investigated. Methods: The MIO-M1 and ARPE-19 cell cultures were incubated in direct contact with DX-PCL
nanofibers and cellular responses were monitored by viability and morphology. The DX-PCL nanofibers were
implanted into the vitreous cavity of rat's eyes and the in vivo biocompatibility was investigated by Optical
Coherence Tomography (OCT) and histology for 10 days. Results and Discussion: The in vitro biocompatibility
test revealed that neither the DX-PCL nanofibers nor the degradation products of the polymer and DX lixiviated
from the systems were cytotoxic to the ocular cells. These cells were capable of proliferating and forming an
organized monolayer. MIO-M1 cells expressed GFAP, demonstrating their functionality. The in
vivo
biocompatibility test indicated that the nanometric drug delivery systems were well tolerated by the ocular tissues,
since they did not alter the architecture of the neurosensorial retina and other ocular structures. Conclusions: The
DX-PCL nanofibers demonstrated high degree of cellular biocompatibility and short-term ocular tolerance. They
represent potential non-conventional pharmaceutical dosage forms for ophthalmologic applications. Ethical
approval: All experiments were performed in accordance with the European Community's Council Directive
86/609/EEC and approved by ethical committees of the René Descartes University (134/2015). Financial Support:
The authors wish to thank the UFSJ (Brazil), CNPq/MCT (Brazil), Fapemig (Brazil) and CAPES (Bolsistas da
CAPES-Brasília/Brazil) for the financial support.
Financial Support
UFSJ (Brazil), CNPq/MCT (Brazil), Fapemig (Brazil) and CAPES (Bolsistas da CAPES-Brasília/Brazil)
Acknowledgments
Academic
Ethical approval
Yes
318
ELUCIDATING THE GUEST-HOST INTERACTIONS AND COMPLEX FORMATION OF
HECOGENIN ACETATE AND BETA-CYCLODEXTRIN BY ANALYTICAL AND
MORPHOLOGICAL TECHNIQUES
YASMIM MARIA BARBOSA GOMES DE CARVALHO
BRUNO DOS SANTOS LIMA
LUCINDO JOSÉ QUINTANS JÚNIOR
Department of Physiology, Federal University of Sergipe, São Cristóvão, SE, Brazil
DANIEL PEREIRA BEZERRA
Gonçalo Moniz Research Center, Oswaldo Cruz Foundation, Salvador, BA, Brazil
CELSO VATARU NAKAMURA
Department of Basic Sciences, State University of Maringa, Maringá, PR, Brazil
FLÁVIO MACHADO DE SOUZA CARVALHO
Departament of Mineralogy and Geotectonic, University of São Paulo, São Paulo, SP, Brazil
THIAGO MENDONÇA DE AQUINO
Laboratory of Nuclear Magnetic Resonance, Institute of Chemistry and Biotechnology, Federal University of Alagoas, Maceió, AL, Brazil
ADLEY ANTONINI NEVES DE LIMA
Departamento f Pharmacy, Federal University of Rio Grande do Norte, Natal, Rio Grande do Norte, Brazil
Introduction: Hecogenin acetate (HA) is steroidal sapogenin-acetylated with pharmacological activities
already been elucidated in the literature, for example: anti-inflammatory, analgesic and gastroprotective, but has
low solubility in aqueous media. Therefore, the formation of inclusion complexes (IC) with beta-cyclodextrin (betaCD) is in favor. Methods: The complexes were prepared by physical mixture (PM) and freeze dryer (FD) in the
molar ratios of 1:1 and 1:2, characterizing them by scanning electron microscopy (SEM), X-ray diffraction (XRD),
mass spectrometry (MS), laser nuclear magnetic resonance (NMR) and docking. Results and Discussion: SEM
analysis of HA, beta-CD and PM in both molar ratios showed square and rectangular-shaped crystals in different
sizes, in the FD in both molar ratios was observed loss of the crystal structure and decrease particle size,
corroborating with XRD results that showed decreased intensity (4,76º) and the appearance of new peaks (17,50°
and 18,66°), suggesting change of crystalline phase. MS and docking results showed interaction between HA and
beta-CD and, through measurement the molecular mass and energy of complex formation, respectively. NMR
analysis confirmed the formation of IC by the FD on the molar ratio 1:2, through chemical shift changes for H-3
and H5 βCD protons that were higher compared with protons located outside the cavity. Conclusions: The
guest-host interactions and complex formation of HA and beta-CD was investigated and confirmed for 1HNMR.
Thus, complexation technique by FD in the molar ratio 1:2 is promising in order to optimize the biological and
physical-chemical characteristics the low solubility substances, as HA, considering as an important step in the
development of novel safer and more effective formulations.
Financial Support
Acknowledgments
We thank Maria Lúcia Vieira Moreno, Cláudio Pereira Figueira and Adriana Lanfredi Rangel for the images of SEM in the Gonçalo Moniz Research Center,
Oswaldo Cruz Foundation, Bahia, Brazil.
319
320
DROPOUT PREVALENCE AND ASSOCIATED FACTORS IN RANDOMIZED CLINICAL
TRIALS OF ADOLESCENTS TREATED FOR DEPRESSION: SYSTEMATIC REVIEW AND
META-ANALYSIS
Adriane Isabel Rohden
Mariana Canellas Benchaya
Roger Santos Camargo
Tais de Campos Moreira
Helena Maria Tannhauser Barros
Maristela Ferigolo
Introduction: Depression currently affects 350 million people. Prevalence among adolescents is 4-5%.
Treatment with antidepressants assists in healing and remission of symptoms, improving life in various aspects.
In the case of abandonment (dropout) of treatment or a clinical trial, the adolescent is no longer followed by the
medical team. We analyzed the dropout rates of randomized clinical trials with depressed adolescents receiving
antidepressant drugs and the factors associated with non-adherence by summarizing this information in a
systematic review and meta-analysis. Method: Articles were retrieved from Medline, Embase, Cochrane, Clinical
Trial, Psycinfo and Web of Science using the MeSH terms "depressive disorder", "randomized trials" and
"adolescents". The evaluation of study quality was performed using the Cochrane Handbook for Systematic
Reviews of Interventions and Jadad scales. Results: The final sample included 50 articles. The overall dropout
prevalence was 23% (95% CI 2027; p < 0.0001). The highest dropout prevalence was associated with
adolescents over 16 years of age (33%), group treated with serotonin norepinephrine reuptake inhibitors (SNRIs)
(45%), and those treated with only medication (15%). Dropout prevalence was most associated with adverse
effects, followed by problems relating to clinical trials and family arbitration. Discussion: Autonomy or substance
abuse may be associated with high prevalence of dropout in adolescents over 16 years. The SNRIs class is not
indicated for adolescents. Cognitive-behavioral therapy associated with pharmacotherapy can change lifestyle,
increase encouragement, and prevent relapse. Adverse effects are most prevalent in the medication with low
selectivity. Conclusion: Older adolescents, SNRIs class and treatment with only medication was showed the
highest dropout. Adverse effects was the factor most associated with non-adherence. This research was financed
by AMTEPA and SENAD.
Financial Support
Secretaria Nacional Antidrogas e Associação Mario Tannhauser de Pesquisa e Ensino
Acknowledgments
Secretaria Nacional Antidrogas e Associação Mario Tannhauser de Pesquisa e Ensino
321
IMPACT OF DISTANCE EDUCATION ON ACADEMIC PERFORMANCE IN
PHARMACEUTICAL CARE COURSE
Agnes Nogueira Gossenheimer
Mauro Silveira de Castro
Mára Lúcia Fernandes Carneiro
Introdution: Many changes have been occurred in the last decade in the health education area, in order
to graduate professionals who can have a humanistic look and can work in Brazilian Public Health System.
Pharmacy Course also has changed due to curricular reforms, including subjects such as Pharmaceutical Care.
The objective of this study was to compare performance of pharmacy students from the Pharmaceutical Care
course, taught in both the Distance Education (DE) and campus-based formats using active methodologies.
Methods: For two semesters, students (n=82) taking the course studied half the course in the DE format and half
face-to-face. Questionnaires were applied at the beginning of the semester aimed to outline the demographic
profile of the students. Their grade in the course was evaluated to determine their performance. Results and
Discussion: The Module 1 (Information on Medications) average in the face-to-face was 7.1225 and in DE was
7.5519, (p=0.117). The Module 2 (Pharmaceutical Services) average in the face-to-face was 7.1595 and in DE
was 7.7025, (p=0.027*). There was difference in learning outcomes in Pharmaceutical Care Course between faceto-face and distant education. Than the Student performance was better in the DE module, indicating DE can be
satisfactorily used in the Pharmacy Program. Conclusions: Significant difference in grade outcomes was found
between pharmacy students who attended a student-centered, active-learning pharmaceutical care course in a
traditional classroom setting and those who attended the same course via Distance Education. Suggesting that
students on both received an equivalent curricular experience, but performance was better in Distance Education.
Although these conclusions should propagate confidence among the pharmacy education community that novel
media are equal and better in effect to traditional methods of teaching.
Financial Support
CNPq
322
ACTIVE METHODOLOGY IN BLENDED LEARNING: AN EXPERIENCE IN
PHARMACEUTICAL CARE
Alexandra Ingrid dos Santos Czepula
Universidade Federal do Paraná (UFPR)
Cassyano Januário Correr
Roberto Pontarolo
Introduction. The aim of this study was to analyze the implementation of an active methodology in a
blended model of education in the teaching-learning process applied to students enrolled in the disciplines
Pharmaceutical Care I and II of the Bachelor of Pharmacy at the Federal University of Paraná. Methods.
Implementation and comparison of results of diagnostic assessments (pre-test) and summative (post-test), based
on Bloom's Taxonomy, multiple choice format. Ethical approval: As required by the Research Ethics Committee
(CEP) / UFPR, this research was submitted to board consideration and was approved under the number 317 676.
Results and discussion. In the discipline Pharmaceutical Care I occurred modifications in the pre-test scores and
the overall post-test with an increase in average between the tests (4.8 to 6.3), with a percentage change of
30.2%, which was statistically significant (p <0.05). It was also observed increases in the average percentage
change in the three levels of Bloom's Taxonomy: level I (29.8%), level II (21.2%) and level III (37.2%). In the
discipline Pharmaceutical Care II occurred modifications in the pre-test scores and the overall post-test with an
increase in average between the tests (4.1 to 5.5), with a percentage change of 34.1%, which was statistically
significant (p <0.05). Similarly, it was observed increases in the average percentage change in the three levels of
Bloom's Taxonomy: the level I (16.7%), level II (39.5%) and III level (37.8%). Conclusion. The active methodology
used in the teaching-learning process had positive results regarding the performance of acquiring knowledge. With
the implementation of this methodology was evaluated improved learning among these students, answering our
question problem. This is the first study that relates Pharmaceutical Care and Blended Learning.
Financial Support
CAPES
Acknowledgments
Students of the degree course in Pharmacy UFPR and the Post Graduate in Pharmaceutical Sciences at
UFPR.
Ethical approval
As required by the Research Ethics Committee (CEP) / UFPR, this research was submitted to board consideration and was approve d under the number 317 676.
323
MEDICATION ADHERENCE IN TYPE 2 DIABETES MELLITUS INSULINIZED PATIENTS
IN A FAMILY HEALTH UNIT IN PONTA GROSSA-PR
Ana Paula Veber
DAYANE BOBATO
GERUSA CLAZER HALILA POSSAGNO
ROSILEA CLARA WERNER
Introduction: Diabetes mellitus (DM) is a non-communicable chronic disease that is increasingly
prevalent in the population. Type 2 DM is characterized by resistance to insulin action, due to environmental and
genetic factors. Adequate control of disease is essential to improve the quality of life of patients, and one of the
main factors to be considered is the adherence to treatment. The aim of this study was to verify the nonadherence of patients with type 2 diabetes mellitus to oral medication and insulin. Methods: The Morisky Scale
modified (eight items) was applied through structured interviews, in home visits to DM2 patients, in treatment with
insulin, registered in a Family Health Unit in the city of Ponta Grossa, PR. Patients were classified adherent to
treatment when they reached the maximum sum of 8 points and nonadherent if the values were below 8 points.
Ethical approval for the study was obtained from the State University of Ponta Grossa Ethical Committee by the
number 890157/2014. Results and discussion: We interviewed 30 insulinized patients, 4 (13.3%) male and 26
(86.7%) female. The results of non-adherence were 75% (23) to insulin therapy and 82% (25) to oral medications.
The patients reported that oral medications are forgotten more easily than insulin, probably due to dosing issues.
Oral medications most often require multiple daily doses, and although insulin administration is uncomfortable for
most patients, it was prescribed only once a day. Conclusion: It was concluded that the rate of non-adherence to
treatment was quite substantial in both oral medications as insulin and this is directly reflected in inadequate
control of blood sugar levels. This leaves users more susceptible to secondary complications of this disease.
Ethical approval
890157/2014 - Universidade Estadual de Ponta Grossa
324
IS IT STRATEGICAL THE LEPROSY IN THE STRATEGIC COMPONENT OF
PHARMACEUTICAL SERVICES? AN ANALYSIS IN TERESINA, PIAUÍ, BRAZIL
André Igor Oliveira Prado
Eliana Elisabeth Diehl
Introduction: Leprosy is one of the diseases included in the Strategic Component of Pharmaceutical
Services, and remains an important public health problem in the North and Northeast of Brazil. To understand the
strategy of Pharmaceutical Services (PS) for the control of leprosy, we describe and analyze the PS chain in the
city of Teresina, Piauí, Brazil. Methods: From a qualitative perspective, the data were collected through
direct/participant observation, semi-structured interviews with managers and coordinators of state and municipal
health programs of control of leprosy, and open ethnographic interviews with people affected by the disease. The
study was approved as ethics (Law n. 824385/CEP/SES-SC). Results and Discussion: The chain of PS in
Teresina is focused on the distribution and delivery of medicines for multidrug therapy schemes in health facilities.
There is little participation of pharmacists in the process, summarized in the steps of programming, acquisition
and distribution of medicines. Medications for the treatment of complications of the disease or of the multidrug
therapy do not get much importance in this chain, with reports of users that is common their lack in services. The
evaluation of adhesion to the treatment and pharmacotherapy follow-up could contribute to a better health care for
these people. Conclusions: The participation of PS in the fight against leprosy in Teresina is timid and lacks of
strategy, requiring a stronger acting in all stages of the drugs supply chain, especially in the dispensation.
Financial Support
Not applicable
Ethical approval
Law n. 824385/CEP/SES-SC
325
MOTIVATIONAL INTERVIEW IN CONTEXT OF PHARMACEUTICAL CARE AND
PATIENTS WITH HYPERTENSION
Carine Raquel Blatt
Camila Zandona Bisognin
Adriane Isabel Rohden
Motivational interview (MI) is a coherent protocol and developed to be a collaborative conversation style
for strengthening a person's own motivation and commitment to change. MI takes a patient-centered approach
could be useful to treatment adherence or to promote changing behavior in chronic life-related diseases like
hypertension. The aim is identify studies in which the pharmacist used MI with patients with hipertension.
METHODS: A systematic review was performed using PubMed, Web of Science and Lilacs database and the
mesh terms "hypertension" and "motivational interviewing". The exclusion criteria were: (1) systematic review (2)
economic evaluation (3) other intervention that MI (3) observational studies. RESULTS: There were 83 articles,
only 25 using motivational interviewing to patients with hypertension. Of these, only 2 MI-based patient interview
was conducted by pharmacist. Both study have aim to improve adherence to antihypertensive medicines. The
first only one interview was done and the outcomes were measure six months later. In second study the
intervention included medication review, an MI-based patient interview and at least two follow-up telephone calls
during a time of 6 months. DISCUSSION: The pharmaceutical intervention using MI technique improved
adherence to antihypertensive medication and reduced blood pressure. The intervention was well accepted both
by the pharmacists and the patients, thereby increasing the likeliness of successful implementation in routine
practice. CONCLUSION: MI has been implemented in mainly areas of health, but there was few studies in which
the pharmacist is carried out this intervention. MI could be effective in the context of pharmaceutical care.
Financial Support
CNPQ - Universal 2014
326
STUDIES ON DRUG INFORMATION: A SYSTEMATIC REVIEW
Carlos Adriano Santos Souza
Laboratory of Pharmacognosy, Department of Pharmacy, Federal University of Sergipe, São Cristóvão, SE, Brazil
Thaís dos Santos Ramos
Fernanda Bispo Rezende
Maria Deisianne de Andrade
Lincoln Marques Cavalcanti Santos
Laboratory of Teaching and Research in Social Pharmacy (LEPFS), Department of Pharmacy, Federal University of Sergipe, São
Cristóvão, SE, Brazil
Fernando Henrique Oliveira Almeida
Wellington Barros da Silva
Introduction: The amount of available information on drugs is boundless, the audience who receives the
information is diversified and offers a wide range of knowledge, and so healthcare professionals have different
needs for information on drugs. Methods: A systematic review was performed in the databases Lilacs and Scielo
using the terms "drug information", "drugs information service", "drugs information center". Subsequently, in order
to supplement and expand the search, the following combinations were used: ("drug information" AND "drugs
information service"); ("drug information" AND " drugs information center"); ("drugs information service" AND
"drugs information center"). Results and Discussion: The initial screening made with the terms drug information,
drugs information service and drugs information center led to the identification of 575 articles, of which 176 were
indexed in more than one database. Among the selected articles, 20 were carried out in Brazil and only one was
held in Costa Rica. Regarding scientific production, 76,1% of the papers were published between 2000 and 2014.
For the design of the study, it was observed that authors named their articles as descriptive (62%; n=13),
transversal and descriptive (14%; n=3), prospective (9%; n=2). Studies described as pilot, case control and
literature review amounted to 3% (n=3). Conclusion: Despite the studies assessed the sources of information on
drugs, they evaluated almost entirely the tertiary health level of attention. Although the selected studies presented
high heterogeneity within their design, they do not have strength as the methodology applied. Given the above,
the incipient publication of articles on the subject is observed as well as the lack of appropriate methodologies for
carrying out the studies.
Financial Support
Coordination for the Improvement of Higher Education Personnel (CAPES).
327
PROFILE OF WARFARIN USERS -RELATED HOSPITAL ADMISSIONS
Christiane de Fátima Colet
Tânia Alves Amador
Isabela Heineck
Introduction: Warfarin is a drug with a low therapeutic index and is associated with adverse events and
preventable hospital admissions. The effectiveness of the treatment depends on several factors. The objective of
this research is to identify factors associated with the occurrence of hospitalizations in patients of primary health
care using warfarin. Methods: A cohort study was conducted for a period of eighteen months with outpatients
from the city of Ijuí/RS. The data was colected by interviews and were supplemented with information obtained
from the medical records of the primary and tertiary attention. The statistical analysis used was the Chi-square
test. The project was approved by the Ethics Committee of the UFRGS, on the report 336.259/2013. Results and
Discussion: 69 patients was accompanied, 55.1% were female, with a mean age of 64.3 ± 13.7 years. The
average weekly dose was 30.69 ± 15,19mg. It was observed that hospitalization was associated with age less
than 65 years (p=0.003), use of more than ten continuous medication (p=0.052), presence of more than four
interactions with warfarin (p=0.0132) and VKORC1 gene polymorphism (p=0.0151). There was no statistical
association of hospitalization with: use of plants (p=0.329), self-medication (p=0.549), smoke (p=0.371),
hypertension (p=0.511), diabetes mellitus (p=0.259), BMI (p=0.705), Time-in-Therapeutic Range (p=0.816), renal
dysfunction (p = 0.421) and liver (p=0.545), polymorphism of CYP2C9 (p=0.391), CHADSVASC score (p=0.778),
sex (p=0.541) and educational level (p= 0,623). Conclusions: The results indicate that health professionals who
treated these patients should pay more attention to young patients on multiple medications with potential drug
interactions to prevent hospitalizations.
Financial Support
FAPERGS
Acknowledgments
PPGCF, FAPERGS, SMS de Ijuí
Ethical approval
336.259/2013
328
Pharmaceutical care for patient with nephrotic and metabolic syndromes: a case study
Cintia Maria Carvalho Ribeiro
Universidade Federal do Maranhão (UFMA)
Maria Luiza Cruz
Universidade Federal do Maranhão (UFMA)
Introduction: Nephropathy is characterized by progressive decline in renal function, resulting in physical,
social and emotional limitations that affect the quality of life of patients. In this context, the pharmacist can help
ensuring adequate pharmacotherapy, safe and effective. The aim of this study was to develop pharmaceutical
care in a user with nephropathy and with a polypharmacy regimen in order to improve their quality of life. Methods:
The study design was descriptive study, observational and qualitative using the SOAP method for monitoring
pharmacotherapeutic interventions. Adherence was assessed by Treatment Adherence Measure (MAT) and the
Beliefs about Medicines Questionnaire (BaMQ), which considers the beliefs, needs and user concerns. To
evaluate the quality of life, we used Brazilian version of SF-36 questionnaire. To evaluate pharmacist's skills to
the consultations we used the Medication-Related Consultation Framework (MRCF). Results and Discussion: We
followed up a female, 48, with nephrotic and metabolic syndrome. There were 20 pharmaceutical consultations
between May and November 2015. The patient used 8 drugs: captopril, cyclosporine, metformin, deflazacort,
fenofibrate, NPH insulin, losartan and simvastatin. The main interventions performed were to monitor water
consumption and drugs administration by a mobile application, counseling on food selection, and the weight
reduction. During pharmacotherapeutic follow up the patient lost weight and laboratory findings were within normal
levels. It was classified as an "adherent" to treatment by MAT and "Greater tendency to adhere" by BaMQ. SF-36
questionnaire showed improvement in mental health and vitality. Pharmacist was considered fully skilled of
communicating with the patient in pharmaceutical care process. Conclusion: The pharmaceutical care was
essential to improve the user quality of life. It is expected to encourage other pharmacists to establish a
relationship of care to their patient.
Financial Support
none
Ethical approval
Research Ethics Committee of the Federal University of Maranhão under No. 1,140,692.
329
Responses to litigation for access to medicines in Brazil and Colombia
Claudia Marcela Vargas-Peláez
Postgraduate Program in Pharmacy, Federal University of Santa Catarina, Florianópolis, Santa Catarina, Brazil.
Marina Raijche Mattozo Rover
Mareni Rocha Farias
Introduction: Litigation has risen as an alternative pathway to access to medicines in the Latin American
region. This study aims to characterize and compare the States' responses to litigation for access to medicines in
both Brazil and Colombia. Methods: The regulations related to the measures taken in response to litigation for
access to medicines between 2000 and 2014 were identified by means of an integrative literature review. This
information was supplemented with semi-structured interviews conducted with representatives of the stakeholders
involved in the phenomenon. Results: The Colombian and Brazilian Executive branches have taken similar
measures in response to litigation for access to medicines. These measures include a change in the definition of
essential medicines, the creation of health technology assessment agencies, the extension of the covered
medicines list, and the restriction of the medicines coverage to specific indications. Although not all respondents
agreed on the measures taken, there is a consensus over the fact that the extension of the list on its own is
insufficient to control litigation for access to medicines. Discussion: The analysis about the measures taken by
Brazil and Colombia, in response to litigation for access to medicines allows inferring that measures focused
solely on the incorporation of new technologies in the health systems' coverage are insufficient to control the
phenomenon. Additionally, despite all efforts in this regard, other aspects, such as appropriate overseeing in order
to make stakeholders comply with their obligations, have also an important role in the rising of litigation.
Conclusion: The Brazilian and Colombian experiences show that the control of litigation for access to medicines
requires the implementation of measures that comprise other aspects factors, such as the values and the
relationships among stakeholders, so as to find a comprehensive solution to the phenomenon.
Financial Support
Claudia Vargas received financial support from the Departamento Administrativo de Ciencia, Tecnología e Innovación, Colciencias from Colombia in the form of a
doctoral scholarship abroad, Call No. 529/2011. The research project was funded by CNPq under the Call No. 41/2013 MCTI / CNPq / CT-Health / MS / SCTIE /
Decit - National Network for Research on Health Policies: Knowledge Production for Recognition of the Universal Right to Health, Line 3 - Monitoring and
analysis of decisions and regulati
Acknowledgments
The authors thank to Francisco Rossi, Maria Cristina Latorre, Luis Guillermo Restrepo, José Julián López from Colombia; Teresa Burgin from Argentina and
Consuelo Silva from Chile, by the support to the research project.
Ethical approval
This research project was assessed and approved by the Research with Human Beings Ethics Committee (CEPSH by its name in Portuguese) of the Federal
University of Santa Catarina under Report No. 712.031/2014.
330
UNLICENSED AND OFF-LABEL USE OF MEDICINES IN AN UNIVERSITY HOSPITAL
Cristiane dos Santos Giuberti
Federal University of Espírito Santo
Gabrielle Costa Nascimento
Pâmella Martins da Costa
Higor Henrique Aranda Cotta
Eliana Zandonade
Introduction: Several drugs used in hospitalized pediatric patients are being prescribed outside the terms
of their product licence that apply for indication, age, dose, or route of administration (off-label) or they are not
specifically licensed for use in children (unlicensed). The risks incurred because of unlicensed and off-label
medication (UOM) are related to inaccuracy rates, contamination during preparation, loss of stability, use of
excipients that have not been tested in children and interactions that can lead to lack of efficacy, adverse drug
effects or even death. This study aims to determine the extension of off-label and unlicensed drugs use in an
university hospital. Methods: A retrospective study, quantitative and qualitative, which analyzed the prescriptions
of Pediatrics sector and Neonates intensive care unit of an University Hospital at Vitória, Espírito Santo, during
30 days. Drugs were coded at the Anatomical Therapeutic Chemical (ATC) and were classified in licensed,
unlicensed and off label based on the following reference sources: the list of licensed drugs of the Brazilian
National Health Surveillance Agency, the database Micromedex® Drugdex® and those available on the literature.
This study was approved by the Ethics Committee on Human Research of Federal University of Espirito Santo
(N. 1141541). Results and Discussion: A total of 716 prescriptions were analysed. The average was 3.99
medicines per prescription. Among the 89 medicines, 42 were classified as off label (47.19%) and 7 as unlicensed
(7.87%). Analgesics, anti-inflammatory and antibiotics for systemic use constituted the most widely prescribed
ATC classes. Metamizole sodium was the most widely prescribed off-label medicine. Conclusions: The large use
of UOM is an usual clinical practice, since there are others studies with high prevalence of use of these drugs in
brazilian hospitals. Measures to rationalize the use of medicines in children must be considered.
Acknowledgments
LESTAT
Ethical approval
Ethics Committee on Human Research of Federal University of Espirito Santo (N. 1141541)
331
EVALUATION OF GERIATRICS CONTENT IN THE CURRICULUM OF PHARMACY OF
BRAZIL COURSES
Daniel Tenório da Silva
Laboratory of Teaching and Research in Social Pharmacy (LEPFS), Department of Pharmacy (LEPFS), Federal University of Sergipe, São
Cristovão, SE, Brazil; Federal University of Vale do São Francisco, Petrolina, PE, Brazil
Fernando de Castro Araujo Neto
Cristovão, SE, Brazil
Sheila Ramos Feitosa
Elisdete Maria Santos de Jesus
Adriana Andrade Carvalho
Pharmacy Department of Lagarto, Federal University of Sergipe, Brazil
Vanessa Alves da Conceição
Divaldo Pereira de Lyra Júnior
Introduction: Advances in health and epidemiological profiles transitions justify the aging population and
the need for geriatrics study in health courses.Thus, the aim of this study was to evaluate aspects of geriatrics
education in Brazil Pharmacy courses. Method: A documentary research was performed to evaluate the geriatrics
content present in Pharmacy courses of Federal Universities in Brazil; then we conducted a cross-sectional study
to measure the experience and views of students on the topic. Results and Discussion: Out of 40 courses, 26
had their curricular matrices available in electronic sites of universities, and only nine had geriatric content. Of
these, seven were in required courses, three electives and in any specific discipline of geriatrics or gerontology
found. Of these, seven were in required courses, three electives and in any specific discipline of geriatrics or
gerontology found. Of the 196 participants, 110 (56.1%) reported having frequent contact with the elderly. Most
were in the fifth year of the course (31.6%) and 80 (40.8%) students said they had owned professional contact
with the elderly. However, 165 (84.2%) said they had no contact with geriatrics themes in their courses. 122
(61.4%) stated that their graduation contributed little or did not contribute to the professional work with the elderly
and 181 (92.3%) agreed or agreed in part that more theoretical-practical geriatrics and gerontology content should
be included in the courses pharmacy. Conclusion: The findings allowed us to visualize the lack of geriatrics and
gerontology content in the curriculum matrices of Pharmacy courses.
Financial Support
This research wassupported by Foundation of Support to Research and TechnologicalInnovation of theState of Sergipe (FAPITEC).
Ethical approval
The study was approved by the Research Ethics Committee (CAAE: 45405715.5.0000.5546) and all students, agreeing to participate in the study were
previously informed about the goals and nature of research, by signing a consent form and clear.
332
DISEASE RISK ASSESSMENT IN CORONARY MEN IN SOUTHERN BRAZIL
Debora de Mello Gonçalves Sant Ana
Universidade Estadual de Maringá
GUERRA-SILVA, NMM
Universidade Estadual do Norte do Paraná (UENP)
SANTUCCI, FS
SANTUCCI, FS
MOREIRA, RC
TASHIMA, CM
MELO, SCCS
TEIXEIRA, LRL
Universidade de São Paulo - Ribeirão Preto.
SANT'ANA, DMG
Introduction: Human health is in focus due to the low demand of this population for health services.
Hypertension is the leading cause of death in Brazil and therefore aimed to evaluate the risk of developing
coronary heart disease in men. Methods: An exploratory quantitative study, conducted with men in the city of
Bandeirantes / PR. Data were collected by spontaneous demand from March 2014 to May 2015, through semistructured questionnaire, clinical examination and blood collection. Results and Discussion: The study included
637 men, mean age was 39 years. Increased incidence of changes with increasing age (p <0.05) for blood
pressure (BP), waist circumference, Body mass index, glucose, total cholesterol, LDL and triglycerides. Of the
252 men who had high BP, 25.39% reported having a previous diagnosis and 43.75% being in treatment. When
analyzed hypercholesterolaemia data, it was found that 34.54% were men with elevated total cholesterol, 19.94%
had elevated LDL, 46.78% presented with low HDL, 36.42% had elevated triglycerides and they had no previous
diagnosis The presence of metabolic syndrome was found in 24.96% men. For the Framingham scale it was
found that 12.56% were intermediate-risk, and 5.49% higher risk for heart disease over the next 10 years. The
scale of ASCVD 553 men had assessed the cardiac risk, and 7.05% had moderate risk. As the average age was
less than 40 years, the high cardiac risk did not appear frequently, as with advancing age the risk factors and the
prevalence of the diseases studied tend to increase. Conclusions: the development of actions in primary care is
needed to diagnose men at intermediate risk and promote health promotion activities so that they do not develop
high cardiac risk.
Financial Support
Financial Support: Project funded by the Araucaria Foundation for the covenants 932/2013 and
Acknowledgments
Acknowledgments: The all nursing students who contributed to the collection of blood.
Ethical approval
Ethical approval: The project was approved by the ethics committee on human research of the Faculty of Pharmaceutical Scien ces of USP under number
707.179.
333
The perception of the tutor on the interaction in a Distance Learning Course on the
Pharmaceutical Care Management
Francisca Maria de Almeida Vargas
Francisca Maria de Almeida Vargas
Mareni Rocha Farias
Introdution: The Distance Education is important in the training of pharmacists of Brazilian Public Health
System (SUS). The Federal University of Santa Catarina (UFSC) and the Brazilian Ministry of Health promoted a
Specialization Course in Pharmaceutical Services Management (2010-2014) for pharmacists who work in SUS. In
the course, management is understood as a technical, political and social process capable of producing results
and occurs in complex contexts involving different understandings, interests and conflicts. The course proposed a
practical activity of prioritization of problems related to pharmaceutical assistance and the preparation of an
Operational Plan as exercise of Situational Strategic Planning. This activity provided the driving ability, leadership
and proactivity in the intervention of concrete problems in reality. The Course also aimed the formation of
networks of collaborative learning and exchange of experiences. In this respect, the tutor has an important
contribution by encouraging interaction that fosters sharing and knowledge building. This study aimed to analyze
the perspective of the tutors about the interaction that has permeated the development of the Operational Plan.
Methods: Qualitative study with data collection by mean three open questions; the technique for data analysis
was collective Subject Discourse, with the help of Software Qualiquantisoft. Results and Discussion: The
speeches formed revealed that the Operational Plan was relevant as a practical activity planning and
management; in view of the tutors and that there was a level of unwanted interaction towards all-all. Conclusion:
The report of the tutors indicates that the Operating Plan, resulted in positive changes in the workplace of
pharmaceutical professionals. Theoretical knowledge about planning and management were meant by reality with
sharing of knowledge and experiences. Approved by the Ethics Committee in Research of the UFSC (252519).
Financial Support
Capes
Ethical approval
Ethics Committee in Research of the UFSC (252519).
334
PHARMACISTS' SKILLS IN CONDUCTING CLINICAL SERVICES IN COMMUNITY
PHARMACIES IN URBAN AREAS OF NORTHEAST BRAZIL
Izadora Menezes da Cunha Barros
Lincoln Marques Cavalcante Santos
Kérilin Stancine Santos Rocha
Tiago Marques Reis
School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil
Felipe Pimentel Paixão
Fernando Henrique Oliveira Almeida
Leonardo Régis Leira Pereira
School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil
Introduction: The pharmacist is responsible for promoting rational medication use and conducting clinical
services in community pharmacies. In order to properly perform such functions, the pharmacist must have
specific technical and scientific training. The aim of this study was to evaluate pharmacists' skills in conducting
clinical services in community pharmacies in urban areas of Northeast Brazil. Methods: A descriptive study was
conducted with pharmacists working in community pharmacies in Aracaju (Sergipe) from July to September 2013.
In the instrument used for data collection, six questions sought to assess pharmacists' knowledge about
dispensing medications. Based on the number of correct answers, it was possible to classify the practitioners'
knowledge as unsatisfactory (0-2 correct answers), regular (1-2 correct answers) and satisfactory (3-4 correct
answers). Results and Discussion: In this study, 221 community pharmacies were visited, and of these, 182 were
excluded. The participants' ages ranged from 20 to 56 years old (average rate: 38), esp females (76.9%). Most
pharmacists (76.9%) presented regular knowledge. The main difficulties described by pharmacists were informing
about medication interactions (41%), correctly advising about medications (28.2%), and explaining medications'
mechanisms of action (28.2%). Moreover, 41% of respondents reported they would need to improve patient
orientation and drug therapy monitoring. These findings suggest the need for investments in training providing
pharmacists with the skills necessary for the proper exercise of their clinical duties. Conclusion: Pharmacists
showed gaps in the knowledge and skills utilized to perform clinical duties. Therefore, it is necessary to devote
supplementation of this knowledge, improving clinical and communication skills, which will also improve public
trust in the prevention and resolution of problems related to medication.
Financial Support
Coordination for the Improvement of Higher Education Personnel (CAPES) and Foundation of Support to Research and Technological Innovation of the State of
Sergipe (FAPITEC).
Acknowledgments
The authors acknowledge pharmaceutical Naiane Dantas and Claudenice Alves for their collaboration in the development this work and all pharmacists who
agreed to participate this research.
Ethical approval
The research project was approved by the Research Ethics Committee of the Faculty of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo
(FCFRP-USP), under the protocol number 114.532 and CAAE 03872312.1.0000. 5403.
335
ASCORBIC ACID USE IN BRAZILIAN CHILDREN
Juliana do Amaral Carneiro Diel
Programa de Pós Graduação em Assistência Farmacêutica, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brasil
Tatiane da Silva Dal Pizzol
Introduction: Ascorbic acid (AA) in case of vitamin deficiency is recommended. It has off-label use as
an effective adjunct in iron replacement therapy. In Brazil, we do not have population-based data on the use of AA
supplementation in children. The goal is to characterize the use of AA in children up to 12 years in Brazil through
the National Survey data Access, Use and Promotion of Rational Use of Drugs (PNAUM). Methods: The
PNAUM is a cross-sectional population-based study. The study population were residents of urban areas. The
data analyzed refers to subjects <12 years. Questionnaires were used answered by the caregiver of the child
during the interview. Information on the use of AA and iron salts was obtained from the question: "In the last 15
days, the 'child's name' used some vitamin, mineral supplement, appetite stimulant and tonic?" Descriptive and
bivariate analyzes were performed, and the variables expressed by relative frequencies and confidence intervals
(CI95%). Results and Discussion: Data of 7528 children were analyzed. AA was the fifth most used drug, with a
prevalence of global use of 1.6% (CI95% 1.3-2.0). In the group of infants <2 years the drug was the second most
used, with a prevalence of 6.9% (CI95% 5.5-8.6), below only acetaminophen, used by 10.1% (CI95% 8.6-11.8). It
was the second most used drug in acute health conditions, with prevalence of 8.9% (CI95% 7.1-11.0) in <2 years
and the fifth most used drug in pre-school (2-5 years), with 3.4% (CI95% 2.5-4.6). AA assists in iron absorption,
which could explain the high prevalence of use in <2 years (6.9%). However, the concomitant use of these two
drugs was less than 1% of cases. Conclusions: AA is a widely used drug in Brazil for children, but it is not yet
clear the reason of their employment. The data available so far dealt mainly on the AA intake in the diet, not a
drug, so the importance of characterizing their use to understand and discuss the importance of its use in children
Financial Support
Secretaria de Ciência, Tecnologia e Insumos Estratégicos Departamento de Assistência Farmacêutica e Insumos Estratégicos Departamento de Ciência e
Tecnologia
Ethical approval
the project was approved by CONEP (398,131 of 9/16/2013).
336
OFF-LABEL DRUG USE IN BRAZILIAN CHILDREN
Juliana do Amaral Carneiro Diel
Tatiane da Silva Dal Pizzol
Introduction: The off-label drug use is the use not approved by a regulatory agency, which is not on the
label, but that does not mean that it is incorrect. In children, it is sometimes necessary to use due to the absence
of formulations and pharmaceutical form specific to this group, or by the lack of evidence about their efficacy and
safety in children. The objective of this study was to check the off-label use of drugs in children ≤ 2 years in
Brazil through data from the National Survey data Access, Use and Promotion of Rational Use of Drugs
(PNAUM). Methods: The analysis data extracted from the PNAUM, population-based cross-sectional study. Data
in this paper refer to individuals with ≤ 2 years of age. It was specific forms answered by the caregiver of the child
present at the time of the interview. Information about the use of drugs was through questions about chronic and
acute drug use by the child. After it was verified the record of these drugs with the National Health Surveillance
Agency (ANVISA), and the Food and Drug Administration (FDA). Performed descriptive analysis and the main
variables were expressed as relative frequencies and their respective confidence intervals. Results and
Discussion: Of all drugs used in chronic diseases, 42.6% (CI95% 31.5-54.6) are approved by the FDA and 65.1%
(CI95% 53.3-75.3) by ANVISA. Of all drugs used in acute diseases, the drugs used were approved by FDA in
53.4% (CI95% 49.8-57.0) are approved by the FDA and 82.9% (CI95% 80.3-85.2) by ANVISA. It is noted that the
drugs for chronic conditions are more used without approval of health authorities probably by the lack of suitable
therapeutic alternatives. Conclusions: Despite the growing incentive for drug research in pediatric populations,
this group remains an orphan population. The industry still prefers to invest in research and development of
pediatric drugs for acute diseases, not showing interest in chronic diseases, causing often use is adapted.
Financial Support
Secretaria de Ciência, Tecnologia e Insumos Estratégicos Departamento de Assistência Farmacêutica e Insumos Estratégicos Departamento de Ciência e
Tecnologia
Ethical approval
the project was approved by CONEP (398,131 of 9/16/2013).
337
DETERMINING CAUSES OF PROBLEMS RELATED TO PHARMACOTHERAPY BY A
PHARMACOTHERAPY FOLLOW-UP PROGRAM FOR PATIENTS WITH CHRONIC
OBSTRUCTIVE PULMONARY DISEASE
JULIANE FERNANDES MONKS DA SILVA
MAURO SILVEIRA DE CASTRO
ANA PAULA DE OLIVEIRA BARBOSA
RAQUEL SOLDATELI VALENTE
EMANUEL VALDEMERI
MARIA ANGÉLICA PIRES FERREIRA
HOSPITAL DE CLÍNICAS DE PORTO DE ALEGRE (HCPA)
LEILA BELTRAMI MOREIRA
Introduction: The pharmacotherapy follow-up (PF) is a methodology of pharmaceutical care scenario
developed to be applied in cases of chronic diseases. Patients with chronic obstructive pulmonary disease
(COPD) need multidisciplinary follow up with an education program in order to understand their condition and
treatment. The objective was to identify causes of problems related to pharmacotherapy (PRP) by a PF program
for patients hospitalized for COPD exacerbation. Methods: a randomized, controlled, not blinded, parallel-group
clinical trial was realized. Inclusion: hospitalized for COPD exacerbation, from July 2012 to May 2014. Exclusion:
poor prognostic comorbidity, communication disability, psychiatric illness, exclusive stay in the emergency.
Patients were randomized to receive intervention pharmaceutical visits during hospitalization and phone calls for a
year after discharge, or as control group, receiving the usual care. Five areas have been established to classify
the causes of PRP: social and economic; related staff; health system; condition of the patient; treatment; patient.
Results: Of the 80 randomized patients, 49 (61.2%) were men, mean age ± SD of 68 ± 9 years, 16 (20%)
illiterate, 51 (63.8%) with a family income of a minimum wage. A smoking history was 74.3 ± 38.7 pack-years.
They identified 576 PRP, 444 (77.1%) in the intervention group and 132 (22.9%) in control. The median (minimummaximum) per patient was 11 (2-30) vs. 3 (0-13) (P <0.001), respectively. The identified causes were related care
team on 297 PRP (44.3%), health care in 134 (20.0%) patients in 129 (19.3%), treatment in 54 (8.1%), social and
economic factors in 50 (7.5%) and patient condition in 6 (0.9%). Conclusion: We identified more PRP in the
intervention group due the PF carried out by the pharmacist. The most prevalent causes were related to the team
and to the health system, which demonstrated the deficiencies in the structure and process of care offered in the
country.
Financial Support
CAPES and Incentive Fund Research of HCPA
Acknowledgments
Thank you for CAPES and Incentive Fund Research of HCPA for the support
received for the research.
Ethical approval
The study was registered in the Brazil Plataform and approved by the Ethical Committee of the Hospital de Clínicas de Porto Alegre (HCPA) (Rio Grande do Sul,
Brazil), with number of protocol 11-0452. All patients provided written informed consent. A copy of the document was given to the participant and one copy
remained with the research team. The study was registered in the Brazilian Registry of Clinical Trials (primary identifier RBR - 5bw2wt).
338
PROFILE OF QUESTIONINGS RECEIVED BY PHONE BY CLINICAL PHARMACY
SERVICE IN A PRIVATE HOSPITAL IN PORTO ALEGRE, RS
Luciana Mello de Oliveira
Professor, Pharmacy course, Universidade de Caxias do Sul, RS
Cintia Pinto Machado
Pharmacist, Hospital Dom João Becker , Gravataí, RS
Dauana Pitano Eizerik
Clinical Pharmacist, Hospital Moinhos de Vento, Porto Alegre, RS
Lisiane Leal
Clinical Pharmacist, Hospital Moinhos de Vento, Porto Alegre, RS
INTRODUCTION: Clinical pharmacists must develop actions aiming medication safe use. In hospital
setting this can be achieved in different manners, including phone contact, but most of the times this practice is
not evaluated. We aimed to quantify and classify the solicitations received through phone calls by the clinical
pharmacy service in the Hospital Moinhos de Vento, Porto Alegre, RS, Brazil. METHODS: Registration of
questions performed through phone calls in the period between 02/05/2014 to 05/05/2014. Data and variables
evaluated were professional requester, type of question performed, and time needed to provide an answer. This
project was approved by the Ethics Comitee of Hospital Moinhos de Vento, CAAE 34036414.7.0000.5330
RESULTS AND DISCUSSION: 689 phone calls were analyzed. Frequent doubts were related to prescription
delivery (15,16%) and medication preparation before infusion (12,57%). Questions about drug compatibility and
drug-drug interactions accounted for 7,82% of the consultations. Difficulty in using electronic prescription system
accounted with 6,57% of consultations. Almost half (46,37%) of the remaining solicitations were miscellaneous;
about a third of these (31,87%) were related to diverse technical instructions about medication preparation and
use, indicating the need of more information and training about medicines. Nursing team made most of the
requests, with a mean of 177 solicitations/month, with nursing technicians and nurses performing, respectively, a
mean of 111 and 67 consultations/month. Total time dedicated to give responses for these 689 solicitations was
17 hours and 29 minutes; each questioning was responded in a mean of 4,56 ±0,48 minutes. CONCLUSION:
This study has shown that there is a need of information among healthcare team in Hospital Moinhos de Vento.
After its performance, the institution developed strategies of education such as trainings to improve knowledge
about medicines.
Financial Support
none
Acknowledgments
none
Ethical approval
Hospital Moinhos de Vento, CAAE 34036414.7.0000.5330
339
The construction of the health needs of patients with severe asthma: a qualitative
study.
Maria Eduarda Nunes
Universidade da Região de Joinville
Maíra Lindomar Pires Toazz
Sandro José Pereira
Universidade da Região de Joinville (UNIVILLE)
Luciano Soares
Universidade da Região de Joinville (UNIVILLE)
Mareni Rocha Farias
Introduction: Needs are important characteristics to understand the use of drugs. Health needs are
understood as a means to achieve a valuable purpose, a result of desired health. This work aimed to analyze the
health needs construction related to pharmacotherapy from severe asthma patients. Methods: Qualitative
research with exploratory approach, which data are based on interviews with patients receiving medicines in a
pharmacy of the Specialized Component of Pharmaceutical Service from the Brazil's Health Ministry. The service
is provides in a municipality of the Santa Catarina state. Interviews were conducted between June and July of
2015 about patient experience on health services and medicines use. Transcripts were analyzed qualitatively
using content analysis through the ATLAS.ti software. The ethics committee from Univille approved the project
(protocol 650.157). Results and Discussion: We interviewed 22 subjects (12 women), mean age 59.5 years (range
18-79 years), mostly white people (n=14), with incomplete primary education, married (n=15). The perception and
knowledge about the own health condition, the presence of symptoms, pain or suffering (related to discomfort and
the limitation of daily activities), the existence or the results of tests or the consideration of medical diagnosis
modulate the formation of the needs expressed. The disease cannot be interpreted independently of
pharmacotherapy, which reinforces the perception of respiratory deficit and its consequences as the essential
factor of individual experience of suffering. This causes the resignification of the need of health, as a means to
reduce the suffering caused by respiratory exacerbations ("visible" or understandable phenomenon), while
diminishes the role of inflammation ("invisible" or incomprehensible phenomenon) in the patient view from the
clinical reality that perceives. Conclusions: Needs impact on the clinical process and the organization of the
health system.
Financial Support
Support Fund Research / Univille; Foundation for Research and Innovation of the State of Santa Catarina.
340
PERCEPTIONS AND PRACTICES OF A TEAM OF A PSYCHOSOCIAL CARE CENTER
ABOUT THE RATIONAL USE OF MEDICINES
MARINA WEIZENMANN
Ministério da Saúde, Secretaria da Gestão do Trabalho e da Educação na Saúde
Tânia Alves Amador
Programa de Pós-Graduação em Assistência Farmacêutica, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brasil
Introduction: The Psychosocial Care Centres (CAPS) are services of daily attention, which enables to
the outpatients, the daily permanence of mental health users in the service and attention to the crisis. Objective:
To describe the concepts that the staff has in relation to the rational use of medicines (URM). Methods: The study
followed a descriptive model, exploratory, qualitative with the workers of the CAPS I from Lajeado-RS and the
sample was intentional. The participants were workers who performed therapeutic intervention directly with
patients, and the groups were composed of the following professionals: doctors, nurses, psychologists, social
workers, occupational therapists, physical educators, nursing technicians, a musician and a person working in a
workshop. The data was collected in focal group to identify attitudes and perceptions. The total number of 15
participants. The discourses were transcribed and the data were analysed by the method of "content analysis".
The project was approved by the Ethics Committee of UFRGS under No. 850924. Results and Discussion: Only
one professional replied that did not know about rational use of medicines. From the discourses of focal groups
emerged 32 categories of initial analysis and of these, it was extracted six final categories: "medicalization of the
society", "the health system structure", "factors that interfere in mental health", "staff conceptions of need for
medication", "suggestions of practices to demedicalization" and "professional qualification and the need to the
health service". Conclusion: The concept of URM was partially showed for the health staff, considering what is
recommended from WHO. The main discourses related the proper dosage and need to use. It was also debated
elements from the health system organization which are necessary to guarantee the URM directly or indirectly,
i.e. the organization of health and care network, the system management and the professional qualification.
Ethical approval
CEP No. 850924
341
PROBLEM PRIORITIZATION IN THE CONTEXT OF PHARMACEUTICAL ASSISTANCE
MANAGEMENT
Monica Cristina Nunes da Trindade
Mareni Rocha Farias
Introduction: The Federal University of Santa Catarina and the Ministry of Health offered two editions
(2010 and 2013) of the distance education Specialization Course in Pharmaceutical Assistance Management to
pharmacists working in the SUS. The course addresses management as a technical, political and social process
capable of producing results, involving different understandings and conflicts. In order to practice situational
strategic planning, a problem-prioritization activity was proposed to the students regarding pharmaceutical
assistance-related problems at their work place. Operational Plans (OPs) were developed so as to tackle the
priority problems (PP). This study aims to comparatively analyze the PPs listed in the two editions of the course.
Method: Data were collected from the OPs uploaded on the learning platform. The problems comprised the 4
categories shown below. Results and Discussion: A total of 1445 OPs were devised in the first edition of the
course, and 1043 in the second edition. Considering the categories, the results of the first and second editions
were respectively: access to medicines 24% and 34%; infrastructure 25% and 26%; human resources 11% and
12%; and pharmaceutical services 40% and 28%. As can be seen, there is a constant of proportionality between
the two editions of the course as regards the categories infrastructure and human resources. In the category of
pharmaceutical services, there was a percentage decrease, while in access to medicines there was an increase.
The analysis considering the municipalities from where these problems stem shows that less than 1% come from
the same municipality, reflecting the policies of information dissemination and care training. Conclusion: The
results show a significant difference in the PPs in the time period under analysis, and issues related to access to
medicines and pharmaceutical services are prioritized over issues of infrastructure and human resources.
Financial Support
Specialization Course in Pharmaceutical Assistance Management, Pharmaceutical Assistance Department and Health Ministry
Acknowledgments
Samara Jamile Mendes, Fabiola Bagatini Buendguens, Andre Felipe Vilvert e Equipe do Curso de Gestão da Assistência Farmacêutica
Ethical approval
Ethical committee of Federal University of Santa Catarina. Protocol number:1.231.402
342
A COMPUTERIZED SYSTEM TO PHARMACOTHERAPEUTIC FOLLOW-UP OF PATIENTS
RECEIVING MEDICINES BY COURT ORDER
SILVIO BARBERATO-FILHO
Pharmacy Graduate Program, University of Sorocaba, Sorocaba, SP, Brazil
CRISTIANE DE CÁSSIA BERGAMASCHI
FERNANDO DE SÁ DEL FIOL
LUCIANE CRUZ LOPES
MARIA INÊS DE TOLEDO
Health Science Faculty, University of Brasília, Distrito Federal, DF, Brazil
EDILMA MARIA DE ALBUQUERQUE VASCONCELOS
Pharmacy Undergraduate Course, University of Sorocaba, Sorocaba, SP, Brazil
TÂNIA REGINA FERREIRA
Pharmacy Undergraduate Course, University of Sorocaba, Sorocaba, SP, Brazil
MARIANA DONATO PEREIRA
Pharmacy Undergraduate Course, Integrated Faculty of the Ribeira Valley, Registro, SP, Brazil
VIVIAN FERRARI DE LIMA SCARANELLO MACHADO
Introduction: Managing health litigation represents a challenge for the Brazilian Public Health System.
However, little research has been conducted that contributes to the management of this need, mainly in relation to
drug efficacy and patient safety. We previously developed a computerized system (Jud Sys System) for recording
and monitoring judicial suits that involve medicines, nutritional therapy, as well as other products and procedures
for health. The aim of this study was to develop a computerized system to assist with and monitor information
related to the pharmacotherapeutic follow-up of patients receiving medicines by court order. Methods: The system
development comprised the following phases: planning, requirements definition, system design and development,
testing, and implementation. The planning phase took into account references on pharmaceutical care methods,
patient counseling techniques, and disease management needs that included compliance, effectiveness, and
medicine safety. After systematizing these parameters, the requirements and the system architecture was
defined. The system testing stage employed simulated clinical cases. Results and Discussion: We opted to have
the system in a Web server, facilitating the process of installing and maintaining. The computerized system
design and development allows for the recording of patient information, their health problems and treatments,
suspected adverse drug events, as well as drugs related problems and pharmaceutical interventions. The
feasibility of implementation of the model in the Brazilian Public Health System considered the constraints faced
by the municipalities, since most of them do not have a pharmacotherapeutic monitoring service. Conclusions:
This computerized system can create a favorable environment to establish pharmacotherapeutic monitoring
services that meet the real needs of both the population and public health system.
Financial Support
The São Paulo Research Foundation (FAPESP)
343
Evaluation of Implementation of a Medication Review Service at an Outpatient Clinic at
the University Hospital in Northeastern Brazil
Tatiane Cristina Marques
Luana Andrade Macêdo
Rafaella de Oliveira Santos Silva
Genival Araújo dos Santos Júnior
Francisco Carlos de Jesus Júnior
Giselle de Carvalho Brito
Introduction: The increase of morbidity and mortality related to pharmacotherapy is considered a
relevant public health issue in many countries. In this context, the implementation of clinical pharmacy services
can solve and prevent drug related problems (DRPs). Thus, this study aimed to evaluate the process in the
implementation of a collaborative Medication Review service between pharmacists and physicians in an
outpatient clinic. Methods: An observational pilot study with longitudinal design was carried out between March
2013 and February 2014. The study population was patients treated at Medication Review service at the
outpatient clinic of a University Hospital in Aracaju-SE, in Brazilian Northeast. Data were collected through tools
developed by the researchers, and patients were assessed for socio-demographic and drug therapy profile.
Pharmaceutical interventions were classified according to the type of degree of acceptance. Results and
Discussion: 146 patients were seen as mean age of 53.4 ± 12.2 years, most females (n=111, 76.0%). The
number of visits per patient ranged from 1 to 5 with an average of 2.1 ± 1.1. Their prescriptions contained 3.3 ±
1.9 drugs and 24.5% of them had five or more medications. The study identified 366 DRPs being most of
necessity (67.5%) and 180 (49.18%) of them were solved. This study also reported 907 pharmaceutical
interventions, of which 734 (80.92%) were intended for patients, being orientations on medication administration
techniques the most frequent (n=208, 28.3%). Moreover, 173 (19.08%) interventions were intended for physicians
and for medical students, being most of them accepted (n=160, 98.26%). Conclusion: Collaborative practice
between pharmacists and physicians was effective in the identification and solving of DRPs and may represent a
strategy to promote the rational medication use.
Financial Support
Coordination for the Improvement of Higher Education Personnel (CAPES/Brazil) and National Counsel of Technological and
(CNPq/Brazil).
Scientific
Development
Ethical approval
This study was authorized by the directors of the participating institutions and approved by the Research Ethics Committee of the Federal University of Sergipe
(CAAE: 11735412.5.0000.5546). The collected data were for the exclusive use of the researchers having secured the confidentiality of information obtained in
accordance with resolution CNS nº. 466/2012
344
EVALUATION OF POSSIBLE RISK FACTORS FOR DRUG INTERACTIONS IN
PRESCRIPTIONS OF RESIDENTS OF LONG TERM CARE FACILITIES IN NORTHEAST
BRAZIL
Vanessa Alves da Conceição
1Laboratory of Teaching and Research in Social Pharmacy (LEPFS), Department of Pharmacy, Federal University of Sergipe, São
Cristovão, SE, Brazil.
Daniel Tenório da Silva
2Federal University of Vale do São Francisco, Petrolina, PE, Brazil.
Vanessa Lima de Santana
Edileide Guimarães Santos
Lincoln Marques Cavalcante Santos
Introduction: the use of multiple drugs is a reality in Long Term Care Facilities. In this context,
polypharmacy is pointed to as the main risk factor for drug interactions and increased mortality rates.
Nevertheless, other risk factors may be determining for the prevalence of possible interactions, which have not
been explored in this scenario. Thus, the aim of this study was to assess possible risk factors for drug
interactions in prescriptions of residents of Long Term Care Facilities. Methods: a cross-sectional study was
conducted in three Long Term Care Facilities in two cities of Sergipe, during 12 months. A sample with 132
elderly residents were assessed by three pharmacists. Potential drug interactions were classified according to the
database Micromedex®. Mantel Haenzel's chi-square test (p<0.05) was used to evaluate the statistical
significance of drug interactions with the following topics: potentially inappropriate medications, therapeutic
duplication and polypharmacy. Results and Discussion: one hundred and twenty-five elderlies were included in
this study with a greater proportion of females (64.4%) and participants' mean ages of 81.8 (standard deviation:
9.6 years). Five hundred and eleven drugs were prescribed identifying 256 potential drug interactions in seventyeight (62.4%) study participants. The elderly has prescriptions of five or more medicines, of potentially
inappropriate medications and of therapeutic duplication presented higher probability for drug interactions.
Conclusion: this study identifies other risk factors, besides polypharmacy, that may affect the prevalence of drug
interactions in residents of Long Term Care Facilities. Future studies should measure the impact of clinical
interventions in reducing the Medication Regimen Complexity Index in elderly assisted in complex Long Term
Care Facilities.
Financial Support
This research was financially supported by the Institutional Program of Extension Initiation Scholarship (PIBIX) and and Foun dation of Support to Research and
Technological Innovation of the State of Sergipe (FAPITEC).
Acknowledgments
We wish to thank all health teams and the elderly residents
in the studied long-term care facilities
Ethical approval
This study was authorized by the directors of the participating institutions and approved by the Research Ethics Committee of the Federal University of Sergipe
(CAAE: 14435913.3.0000.5546). The collected data were for the exclusive use of the researchers having secured the confidentiality of information obtained in
accordance with resolution CNS nº. 466/2012
345
PHARMACEUTICAL CARE FOR A PATIENT WITH RHEUMATOID ARTHRITIS AND
HYPERTENSION IN A POLYPHARMACY REGIMEN: A CASE STUDY
Verônica Maria da Silva Eidan
Federal University of Maranhão
Maria Luiza Cruz
Federal University of Maranhão
Introduction: Rheumatoid arthritis is an autoimmune disease characterized by inflammation of synovial
tissues and extra articular manifestations. Hypertension is defined by high and sustained levels of blood pressure
above 140/90 mmHg. The treatment involves both educational and measures mainly pharmacotherapy. It is in this
follow-up care that the pharmacist is included, to ensure appropriate, safe and effective pharmacotherapy.
Methods: The study design was descriptive study, observational and qualitative using Dader method for
monitoring pharmacotherapeutic follow up. To assess adherence, we used Treatment Adherence Measure (MAT)
and the Beliefs about Medicines Questionnaire (BaMQ). To evaluate the quality of life, we used SF-36
questionnaire. The pharmaceutical skills to carry out the consultations were evaluated. Results and Discussion:
Female, 59, with rheumatoid arthritis and hypertension. The study took place between July / 2015 and February /
2016. The polypharmacy regimen comprises: hydroxychloroquine, prednisone, naproxen, ranitidine, ibandronate
sodium, cholecalciferol, calcium carbonate, golimumab, potassium losartan and hydrochlorothiazide. The
interventions included self-monitoring daily water consumption, tracking record blood pressure, pill organizer for
easy access to needed medications and encouraging the practice of daily stretching. There was a reduction in
blood pressure levels, the activity of rheumatoid arthritis and weight loss. She has "greater tendency to adhere to
treatment" according to BaMQ, was classified as "adherent" to treatment through the MAT and improved in all
domains of the SF-36 questionnaire during follow-up. The pharmacist was considered fully skilled in providing this
service to the user. Conclusion: The pharmaceutical care process was essential to improve the user's quality of
life, which demonstrates the importance of the pharmacist in patient care.
Ethical approval
The study was approved by Research Ethics Committee of the Federal University of Maranhão, No.
1,165,149.
346
347
ATORVASTATIN INHIBITS INTERLEUKIN 17A (IL-17A) SECRETION IN PERIPHERAL
BLOOD MONONUCLEAR CELLS FROM RHEUMATOID ARTHRITIS PATIENTS
Adson Belém Ferreira da Paixão
Laurindo Ferreira da Rocha Júnior
Moacyr Jesus Barreto Melo Rêgo
Angela Luzia Branco Pinto Duarte
Introduction Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by joint damage
and comorbidities at systemic level. Many cells and molecules are responsible for the inflammatory response that
leads to joint tissue injury. Interleukin 17A (IL-17A) is a cytokine elevated on rheumatoid arthritis patients that acts
elevating other pro-inflammatory cytokines such as IL-6 and TNFα. It also activates osteoclasts, fibroblasts and
chondrocytes, leading to bone destruction, collagen deposition and fibrosis. RA treatment uses association of
anti-inflammatory drugs to bring relief in symptoms and improve quality of life. Atorvastatin (ATV) is a statin and
has inhibitory action in cholesterol synthesis pathway providing an improvement in lipid profile. A better clinical
picture has been observed on RA patients treated with ATV. However, these mechanisms are not clear. This
study evaluates ATV effects on IL-17A secretion in peripheral blood mononuclear cells (PBMC) from RA patients.
Methods 17 patients were diagnosed with RA according to American College of Rheumatology (1987). PBMC
were purified from whole blood using density gradient method (Ficoll-Paque Plus) and cultured at 106cells in
RPMI medium supplemented with bovine fetal serum (10%). Cells were stimulated with phorbol myrystate
acetate+ionomicin, exposed or not to ATV (110μM) and incubated for 48h. Supernatants were collected and IL17A dosed through Cytometric Bead Array Th1/Th2/Th17 kit. Data were analyzed using Wilcoxon test and
differences considered significant when p<0.05. Results and discussion ATV significantly reduced IL-17A levels
in PBMC culture from RA patients at 0.01μM (p=0.0049), 0.1μM (p=0.042), 1μM (p=0.0015) and 10μM (p=0.0002)
doses when compared to stimulated control. Conclusions Our findings showed that ATV therapy inhibits IL-17A
on RA PBMC culture. Further experiments are necessary to quantify other cytokines related to RA pathogenesis.
Financial Support
CAPES, FACEPE, INCT_if, PROPESQ_UFPE
Ethical approval
CEP/CCS/UFPE Nº 339/10
348
A STUDY ON GASTROPROTECTIVE AND ANTISECRETORY EFFECTS OF ETHANOLIC
EXTRACT FROM LEAVES OF Spondias mombin L.
ALMIR GONÇALVES WANDERLEY
UNIVERSITY FEDERAL OF PERNAMBUCO
Samara Alves Brito
Cynthia Layse Ferreira de Almeida
Jessica Carla Bezerra do Nascimento
Allison Rodrigo da Silva Oliveira
Isabela da Silva Barbosa
Maria Joanellys dos Santos Lima
Lécio Leone de Almeida
UNIVERSITY RURAL FEDERAL OF PERNAMBUCO
Germana Freire Rocha Caldas
UNIVERSITY FEDERAL OF MARANHÃO
INTRODUCTION: Spondias mombin L, cajazeira, is popularly used for stomach pain relief. The
purpose of this study was to investigate the gastroprotective and antisecretory activities of the ethanolic extract
from leaves of S. mombin (SmEE). METHODS: The leaves were collected in Cajazeirinhas, PB and identified by
experts of the Agronomic Institute of Pernambuco, and the voucher specimen received the number #89987. The
dehydrated leaves were macerated with ethanol (100 g/100 mL) yielding 7.3%. The in vivo experiment were
performed with male Wistar rats (2-3 months) coming from vivarium of the Department of Physiology and
Pharmacology of UFPE, maintained at controlled conditions and received water and ration (Presence®) ad
libitum. The gastroprotective activity was evaluated by inducing ulcer with ethanol and the animals were pretreated (6 animals/group, p.o.) with lanzoprazole (30 mg/kg), NaCl 0.9% (10 mL/kg) and SmEE (50, 100 e 200
mg/kg). The ulcers were quantified by planimetry (ImageJ®), computed in mm2 and analyzed by histopathology.
The antisecretory activity was evaluated by the ligation of pylorus (6 animals/group) using pre-treated animals
(i.d.) with SmEE (100 mg/kg), NaCl 0.9 % and ranitidine (60 mg/kg). Differences between groups were analyzed
by ANOVA and Dunnett's post-test (p<0.05). The experimental protocol was approved by CEUA of UFPE.
RESULTS AND DISCUSSION: The SmEE (50, 100 and 200 mg/kg) caused a significant decrease of the
ulcerated area to 175.0 ± 17.78, 22.20 ± 3.22 and 22.35 ± 2.14 mm2 respectively, when compared to the control
group (229.8 ± 11.01 mm2). Through pylorus ligature, it was found that the SmEE (100 mg/kg) reduced the gastric
secretory volume (0.82 ± 0.08 mL), when compared to the saline group (1.12 ± 0.06 mL). This extract inhibited
significantly the total gastric acidity and increased the pH of the gastric juice. CONCLUSION: The ethanolic
extract of S. mombin presented antiulcerogenic and antisecretory activities.
Financial Support
CAPES
Acknowledgments
CAPES
Ethical approval
PROCESS NUMBER 23076.013615/2015-11
349
CHEMICAL COMPOSITION AND PSYCHOPHARMACOLOGICAL PROFILE OF Citrus
aurantium L. ESSENTIAL OIL IN MICE EXPOSED BY INHALATION
Amanda Zamboni
Janaína Lucas de Oliveira Salomón
Ana Cláudia Fagundes
Maíra Kerpel dos Santos
Adriana Nunes Wolffenbüttel
Mirna Bainy Leal
Renata Pereira Limberger
Introduction: Available treatments for anxiety have been associated with significant adverse effects or
are not effective for some patients. Therefore, the evaluation of new compounds with potential anxiolytic activity
can bring new alternatives for more effective and safe treatments. Methods: Commercial sample of Citrus
aurantium L. essential oil obtained from leaves (EOCa) was analyzed by gas chromatograph coupled to mass
spectrometer. Groups of 8-12 swiss, adults, male mice were subjected to inhalation exposure. Tween 80 1% or
EOCa 10% were nebulized for 30 min in an inhalation chamber. For psychopharmacological analysis, light-dark
(LDT), locomotor activity (LA) and tale suspension (TST) tests were performed. In LDT, control groups were
treated intraperitoneally (i.p.) with saline and diazepam (2 mg / kg) and after LTD, they were placed in LA box. In
TST control groups, they were treated i.p. with saline and imipramine (20mg / kg). Results were analyzed by
ANOVA / SNK. Results: Liquid EOCa presented as major constituents limonene (2.6 %), linalool (22.4%), linalyl
acetate (28.0%) and myrcene (7.8%), proving its legitimacy and purity. Linalool vaporized concentration was 1%.
LA was not altered after treatment with EOCa (p = 0.111). In LDT, there was no significant difference in the
number of crossings (p = 0.479), diazepam group showed significant difference in latency to the first intersection
(p = 0.023) and the time spent in the light compartment (p <0.001). In TST, there was no significant difference
between the control groups and treated with EOCa (p> 0.05). Imipramine showed a significant difference (p
<0.001) in immobility time compared to the saline group. Conclusions: Results did not indicate anxiolytic or
antidepressant activities of EOCa for vaporized concentration of 1% linalool, counteracting to the activity shown
by a previous study of 3% vaporized linalool. The results of these analyses will serve as a guide for future
studies.
Financial Support
CAPES, CNPq, FAPERGS, PROPESQ/UFRGS
Ethical approval
CEUA/UFRGS 18717
350
GLYCOSYLATION WITH OLinked βNacetylglucosamine (OGlcNAc) INDUCES
PRODUCTION OF SUPEROXIDE ANION IN THE VASCULATURE.
Andressa Almeida Franco
Universidade Federal de Mato Grosso (UFMT)
Hellida da Costa Alves Teixeira
Fernanda Regina Casagrande Giachini
Victor Vitorino Lima
Background: Reactive oxygen species (ROS), such as superoxide anion (●O2), which is formed by an
electron reduction of molecular oxygen by enzymes such as NADPH oxidase, can modulate intracellular signaling
pathways and generate others sources of ROS. The NADPH oxidase is an important source of ●O2 in vascular
cells, which has its subunits attached to the membrane (Nox-1Nox-4 and, p22phox) and cytosolic subunits
(p47phox and p67phox). Whereas important proteins with vascular function are targets of O-Linked βN
acetylglucosamine (O-GlcNAc) post-translational modification, it is not known the association between ROS
production and O-GlcNAc modulation in the vasculature. The present study aims to evaluate whether the increase
in O-GlcNAc positively modulates ROS through the activation of NADPH oxidase in the vasculature. Methods:
Aortas from 14-16 weeks-old male Wistar rats, were incubated with vehicle or PUGNAc (100 µM), a selective
inhibitor of OGA, for 12 or 24 hours. ●O2 production was evaluated by using dihydroethidium stain and subunits
of NADPH oxidases subunits where evaluated by western blot. Results and Discussion: We found that the
production of ●O2 was increased in aortas treated with PUGNAc, both for 12 or 24 hours, when compared with
control. PUGNAc incubation also resulted in increased expression of Nox- 1, Nox-4 and p22phox subunits.
Additionally, translocation of p47phox subunit was observed, contributing to NADPH oxidase activation.
Conclusions: Our results suggest that augmented levels of O-GlcNAc contribute to NADPH oxidase activation,
favoring the production of ● O2-. This finding may represent an additional mechanism to explain the frequent
association of O-GlcNAc, ROS and vascular dysfunction.
Financial Support
UFMT, CNPq, FAPEMAT, CAPES.
Ethical approval
protocol 013/2013
351
IL-10 EXOGENOUS PROTECTS VASCULAR FUNCTION IN HYPERTENSIVE ANIMALS
Anielly Almeida Rocha
Victor Vitorino Lima
Fernanda Regina Giachini
Introduction: Interleukine (IL) -10 acts limiting and terminating the inflammatory response by inhibiting a
variety of immune parameters. Recent studies show an inverse relation between IL-10 and Angiotensin (Ang II)
during hypertension. We hypothesized that IL-10 limits increased vascular reactivity to contractile stimuli in
arteries from Ang II hypertensive mice. Methods: Osmotic mini pumps (0.25 μl/hour 14 days - model 1002, Alzet
Co., CA) were implanted subcutaneously to continuously infuse Ang II (1.8 mg/14 days), IL-10 (10 μg/14 days) or
vehicle (saline), for 14 days. Wild-type (WT) mice were infused with Ang II (90 ng/min) or vehicle. Additionally,
some mice were infused with exogenous IL-10 (0.5 ng/min), in the presence or absence of Ang II infusion. After
14 days of infusion, aortic rings were mounted in a myograph and concentration response curve to phenylephrine
(PE) were evaluated. Results After 14 days, plasma IL-10 levels (pg⋅mL) were smaller in Ang II- infused mice,
compared to vehicle-infused mice (4.9±2.3 vs. 10.6±1.5, respectively). Exogenous IL-10 infusion increased
plasma IL-10 levels both in vehicle-infused (18.3±2.8) and in Ang II- infused mice (4.4±2.3). Ang II-infusion
increased PE-induced contraction (Emax; % KCl) in aortas, compared to control (212±9 vs 175±9). Augmented
PE-induced contraction was reduced in aortas from mice co-infused with exogenous IL-10 and Ang II (167± 5%),
as well as in aortas from mice infused exclusively with exogenous IL-10 (139±3%). Conclusion: These results
demonstrate that IL-10 counteracts vascular dysfunction associated with hypertension. Strategies to enhance IL10 levels during hypertension may enhance the benefits provided by regular treatments.
Financial Support
Loreal, CNPq, CAPES e FAPEMAT
Acknowledgments
NO
Ethical approval
23108.010943/14-0
352
FOR ANTIPLASMODIAL ACTIVITY OF Rhinella marina FROM SOUTHERN AMAZON
Bryan Wender Debiasi
Domingos de Jesus Rodrigues
Institute of Natural, Humanities and Social Sciences, Federal University of Mato grosso, 78557-267, Sinop, MT
Gerardo Magela Vieira Júnior
Departament of Chemistry, Federal University of Piauí, 64049-550, Teresina, PI
Bruno Antonio Marinho Sanchez
Institute of Health Sciences, Federal University of Mato grosso, 78557-267, Sinop, MT
Stela Regina Ferrarini
Institute of Health Sciences, Federal University of Mato grosso, 78557-267, Sinop, MT
Introduction: The 2015 Nobel Prize in Physiology or Medicine was awarded to three scientists who
made pioneering discoveries of drugs currently used to treat neglected tropical diseases. A portion of the award
was to Youyou Tu, guided by ancient Chinese texts, isolated a component from the plant Artemesia annua, which
proved to be highly effective against the malaria parasite. The use of animals, plants, fungi and bacteria are
important sources of biologically active substances. Our objectives has been conducted bioprospecting in
extracts and pure compound from R. marina venom, occurring in the Southern Amazon in toxic, antiplasmodial
and cytotoxic activities Methods: Samples of toad venom of R. marina were dried, powdered and extracted three
times with CHCl3/MeOH. The extract from R. marina venom was chromatographed on Sephadex LH-20 column,
yielding the compound 1. Results and Discussion: Extract and 1 were evaluated in human hepatoma (HepG2) and
normal kidney glomerular (BGM) cell lines using the colorimetric MTT and Neutral Red assays. Chloroquine (CQ)
resistant W2 strain of P. falciparum was used for in vitro blood stage culture to test the antiplasmodial efficacy of
toad venom extract and 1. The telocinobufagin (1) showed a high reduction of parasitemia in vitro. Venom extract
and 1 showed no cytotoxic activity against HepG2 and BGM cells in MTT assay. Besides, when was analyzed
the cytotoxic activity of the compounds of Neutral Red assay, it was observed that all samples showed high
LC50 values in the HepG2 and BGM cells, not being thus cytotoxic to both cell. Conclusions: In summary, toad
venom extract showed high toxicity against A. salina. The extracts of R. marina venom and 1 isolated from R.
marina showed pronounced lethal effects in P. faciparum chloroquine resistant strain and low cytotoxic effect,
highlighting toad parotoid gland secretions as a promising source of novel lead antimalarial compounds. Lipid
Nanocapsules Core are being developed
Financial Support
CNPq e UFMT
Acknowledgments
CNPq, FAPEMAT and UFMT
353
CEFAZOLIN RAT'S PLASMA PROTEIN BINDING DEPENDECY ON DRUG
CONCENTRATION IN OBESE RATS
Faculty of Pharmacy, Federal University of Rio Grande do Sul
Eduardo Célia Palma
João Victor Laureano
Bibiana Verlindo de Arújo
Teresa Dalla Costa
Introduction: Cefazolin (CFZ) is the first choice antimicrobial for prophylaxis of surgical wound infection
in bariatric surgery. Obesity can reduce CFZ free (pharmacologically active) subcutaneous concentrations in
rodents (data no published). Furthermore, it has been shown that CFZ binding to human's plasma protein is
concentration-dependent. This study aimed to investigate CFZ protein binding in obese and non-obese rat plasma
in the concentration range observed following i.v. bolus dose (30 and 45 mg/kg) to Wistar rats. Methods:
Experiments approved by UFRGS Ethics in Animal Use Committee (25463). Obesity was induced by
hypercaloric diet. Polled plasma from obese and non-obese male Wistar rats were spiked with CFZ solution to
obtain final concentrations of 15, 30, 75, 150, 300 and 600 µg/mL. After incubation, aliquots of plasma were
transferred to a Millipore Centrifree® YM-30 device and centrifuged for 30 min at 1.000 g, 21 ± 1 ºC. The filtrate,
following a cleanup procedure, was used for quantifying CFZ by a validated HPLC/UV method. Quantification of
CFZ total plasma concentration was conducted using the same cleaning up procedure in incubated plasma,
previous to ultrafiltration. Experiments were conducted in duplicate. Results and Discussion: CFZ binding to rat´s
plasma protein showed concentration-dependency, varying from 73.3% (15 µg/mL) to 31.5% (600 µg/mL) in both
studied groups. There was no significant difference between CFZ binding to plasma protein of obese and nonobese rats (p < 0.05). The mathematical modeling of free vs. bound concentrations using Scientist® allowed
determining a dissociation constant of 79.9 µg and a number CFZ binding sites to plasma proteins of 0.4.
Conclusion: Obesity did not affect CFZ protein binding to rat's plasma. The modeling of plasma protein binding
dependency on CFZ concentration will allow the construction of a more adequate model to describe the drug free
interstitial concentrations in non-obese and obese subjects.
Financial Support
PPSUS/FAPERGS 2013 (#1298-255/13-8).
Acknowledgments
Financial support from PPSUS/FAPERGS 2013 (#1298-255/13-8).
Ethical approval
UFRGS Ethics in Animal Use Committee (25463)
354
PHARMACOKINETICS AND SUBCUTANEOUS PENETRATION OF 2 g AND 3 g OF
CEFAZOLIN IN MORBIDLY OBESE PATIENTS
Eduardo Celia Palma
Pharmaceutical Sciences Graduate Program, Federal University of Rio Grande do Sul, Porto Alegre - RS, Brazil.
Nelson Guardiola Meinhardt
Center for Obese Class III Care, Nossa Senhora Conceição Hospital (HNSC), Porto Alegre - RS, Brazil.
Isabela Heineck
Maria Isabel Fischer
Airton Tetelbom Stein
Center for Obese Class III Care, Nossa Senhora Conceição Hospital (HNSC), Porto Alegre - RS, Brazil.
Teresa Dalla Costa
INTRODUCTION: Surgical site infections are common after bariatric surgery. Antibiotic prophylaxis
protocol recommends the same cefazolin (CFZ) dose (2 g) for obese and non-obese patients. A 3 g dose has
been suggested for morbidly obese patients based on Monte-Carlo simulations. OBJECTIVE: To evaluate the
pharmacokinetics (PK) and subcutaneous distribution of CFZ after 2 g and 3 g single i.v. bolus dose to morbidly
obese patients. METHODS: Patients undergoing laparoscopic gastric bypass were included in the study. Before
incision, patients received CFZ intravenously. Blood and abdominal subcutaneous microdialysate (CMA 66 probe,
30 mm) samples were collected up to 4 h post-dose. Microdialysis probes were calibrated by retrodialysis. Total
and free (ultrafiltration, Millipore Centrifree®) CFZ plasma levels, and free tissue levels were assayed by validated
LC/UV method. PK profiles were evaluated by non-compartmental analysis (Phoenix®) and parameters compared
by Student´s t test. RESULTS AND DISCUSSION: Patients receiving 2 g (n = 3, BMI 46.7 ± 2.2) or 3 g (n = 4,
BMI 42.7 ± 4.3) of CFZ. Total plasma PK parameters after 2 g dose were: t1/2 of 3.4 ± 2.1 h, AUC0∞ of 556.0 ±
342.2 ug.h/mL, CLtot of 6.8 ± 2.0 L/h/kg, and Vdss of 25.5 ± 8.8 L. No statistical difference was observed in t1/2,
CLtot and Vdss after 3 g dose (p > 0.05). A 50% increase on CFZ dose increased free plasma exposure in 59%.
Free tissue parameters after 2 g and 3 g were, respectively: t1/2 of 2.7 ± 1.0 h and 1.8 ± 1.0 h, AUC0∞ of 124.6
± 88.1 ug.h/mL and 242.2 ± 60.6 ug.h/mL, MRT of 4.4 ± 1.9 h and 3.3 ± 1.9 h. CFZ free tissue penetration
(AUCtissue,free/AUCplasma,free) were 0.60 ± 0.33 and 0.84 ± 0.58, respectively. CONCLUSION: Preliminary
data showed higher subcutaneous CFZ penetration after 50% increase in dose. The hypothesis under
investigation is whether this is due to saturation of plasma protein binding.
Financial Support
PPSUS/FAPERGS 2013 (#1298-255/13-8).
Ethical approval
Ethics in Research Committee approval from UFRGS (572-000) and HNSC (12-058). UFRGS (572-000) e GEP-HNSC (12-058).
355
INHIBITION ASSESSMENT IL-17A IN SUPERNATANT CULTURE FOR A NEW
OXAZOLIDINE IN ASTHMATICS
EUDES GUSTAVO CONSTANTINO CUNHA
GABRIELA SOUTO VIEIRA DE MELLO
JOSÉ ÂNGELO RIZZO
VALÉCIA DE CASSIA MENDONÇA DA COSTA
RENATA VIRGÍNIA CAVALCANTI SANTOS
MARIA DO CARMO ALVES LIMA
IVAN DA ROCHA PITTA
Currently, about 300 million of people worldwide suffer from asthma, a chronic inflammatory disease
that affects the airways of individuals of different ages. It is noted that, in some instances, asthma shows
increased levels of interleukin 17 (IL-17A). Because patients present an increased refractoriness to inhaled
corticosteroids, the development of new substances to improve the quality of life of these patients is essential.
The oxazolidines are compounds of recognized anti-inflammatory activity and have been tested against
respiratory diseases nature. To evaluate the immunomodulatory action of LPSF-NB13 oxazolidine in the
decreasing IL-17A levels in peripheral blood mononuclear cell (PBMC) of adults with moderate to severe asthma.
At the Pneumology ambulatory of Clinics Hospital-UFPE, was collected blood from 5 patients to analyze the
effect of LPSF-NB13 on IL-17A. PBMCs were isolated from blood by Ficoll centrifugation, then they were
stimulated with PMA and ionomycin and finally treated with LPSF-NB13, in concentrations of 1, 10 and 100 μM,
and methylprednisolone (100μM) as a reference drug. IL-17A levels were quantified by ELISA in the supernatants
collected after 48 hours. The synthetic compound showed a reduction of IL-17A levels in patients in 1μM (median
91.88 pg/ml, max 370.06 pg/ml, min 2.12 pg/mL), 10μM (median 100.15 pg/ml, max 287.31 pg/ml, min 1.84
pg/mL) and 100μM (median of 89.59 pg/ml, max 196.27 pg/ml, min 38.74 pg/mL) doses when compared to
stimulated condition. IL-17A is considered closely related to the disease development of asthma, since its
participation is so active in the inflammatory processes . Thus lower levels of this cytokine may lead to an
improvement on quality of life of these patients. This work suggests that the oxazolidine LPSF-NB13 has a
considerable therapeutic potential for the disease in question.
Financial Support
INCT_if , CNPq, FACEPE and CAPES
Ethical approval
711.514/CCS/UFPE
356
EFFECTS OF ESPECÍFICO-PESSÔA ON BIOCHEMICAL PARAMETERS OF MALE
WISTAR RATS
Fabiana Sari Ferreira
Fernanda Coleraus Silva
Alana Meira Reichert
Carla Brugin Marek
Ana Maria Itinose
Assistence Center in Toxicology (CEATOX), Hospital University of Western Paraná (HUOP)
Priscila Da Caz
Suélyn Koerich
INTRODUCTION: The Específico-Pessôa is a hydroalcoholic extract produced by a root of a plant
popularly called 'cabeça de negro', used in folk medicine for treatment of snakebites which occur frequently in
tropical and subtropical countries. Some studies show that is efficient against Bothrops atrox. venoms. However,
the safety of this product is not proven in a healthy organism. Therefore, this study was designed to investigate
some effects of Específico-Pessôa on biochemical parameters of a healthy physiological system. METHODS:
Male Wistar rats were divided in groups. The hydroalcoholic extract Específico-Pessôa was administrated by
gavage at a dosage of 0.25 and 0.75 mg/Kg body weight. The animals were wacthed daily. At the end of this
study, the rats were anesthetized for blood collection. In sequence, they were euthanasied and the internal organs
were removed, weight and gross lesions were observed. The blood was used to evaluate biochemical parameters.
RESULTS AND DISCUSSION: It was observed changes in cholesterol levels and a decrease in the activity of
aspartate aminotransferase (AST). There was also gross lesions in the kidneys of some animals treated with low
dose of the alcoholic extract. Animals treated with the extract, showed agitation, drowsiness and grooming.
These effects show that Específico-Pessôa acts on a physiologic organism by a complex mechanism involving
different pathways and different systems by altering kidney function in an unclear manner. CONCLUSION: More
detailed studies should be conducted to understand the toxic mechanisms involved. However, the available data
indicate that this product is active on the physiological system, suggesting caution in the use of this product,
even at low dose as recommended by the label.
Financial Support
Higher Education Personnel Improvement Coordination (CAPES) and Foundation Araucaria (FA).
Acknowledgments
This work was supported by Laboratory of Cellular Toxicology and The Master's program in Pharmaceutical Sciences from University of Western Paraná.
Ethical approval
Ethics Committee in the use of animals (CEUA) from the State University of West Paraná (Number:
054/11).
357
EVALUATION OF HIPPOCAMPAL DISTRIBUTION OF UNBOUND QUETIAPINE BY
BRAIN AND BLOOD MICRODIALYS IN WISTAR RATS
Fernando Carreño
Carolina de Miranda Silva
Karina Paese
Stela Maris Kuze Rates
Teresa Dalla Costa
INTRODUCTION: The ability to deliver drugs effectively across the blood brain barrier (BBB) is a
challenge when developing new central nervous system (CNS) treatments. Quetiapine (QTP) variable response in
schizophrenia treatment might be related to the drug's transport mechanisms across BBB. Brain microdialysis
(MD) allows to investigate the pharmacokinetics (PK) of drugs with the advantage of providing free
pharmacologically active concentrations at the site of action. OBJECTIVE: To investigate the PK of unbound
QTP in plasma and its hippocampal (HIP) distribution by MD. METHODS: Experiments approved by UFRGS
Ethics Committee in Animal Use (25737). Male Wistar rats were submitted to surgery for MD HIP probe (CMA
12, 3 mm) implantation (A:-5.20 mm; L:+4.80 mm; V:-4.50 mm), and MD probe (CMA 20, 4 mm) implantation in
the jugular vein. Forty eight hours later, unbound HIP and blood concentrations were determined after a single
QTP 10 mg/kg i.v. bolus dose (n = 4). Probes recovery were previously determined by retrodialysis in vivo. MD
samples were collected every 30 min up to 5 h after dosing. QTP was determined in microdialysate samples by a
validated LC/UV method. PK parameters were determined by non-compartmental analysis (NCA) using the
Phoenix® v. 64 (Pharsight®). Free brain penetration factor (fT) was determined as the ratio between AUC05h,free,HIP/AUC0-5h,free,plasma and it was used as a measure of QTP brain penetration. RESULTS: In vivo
probe recovery was 22.0 ± 2.8% for brain and 34.7 ± 3.2% for blood. QTP showed an AUC0-5h,free,plasma of
1370 ± 125 ng¿h/mL, t1/2 of 2.6 ± 1.5 h with a mean clearance of 1.15 ± 0.15 L/h/kg and a volume of distribution
of 3.5 ± 1.74 L/kg. HIP exposure to QTP was 2248 ± 499 ng¿h/mL with t1/2 of 6.2 ± 2.1 h. The fT was 1.64 ±
0.15. CONCLUSIONS: A fT higher than one indicates that influx transporters are involved in QTP penetration
across BBB. The identification of influx the transporter(s) involved remains to be determined.
Financial Support
Master scholarship from CNPq/Brazil
Ethical approval
CEUA/UFRGS #25737
358
EVALUATION OF CENTRAL ACTIVITY OF HIDROETANOLIC EXTRACT FROM JUSTICIA
PECTORALIS (HEJP) IN AN ANIMAL MODEL OF ALZHEIMER'S DISEASE (AD)
Glenda Laíssa Oliveira de Melo Candeia
Department of Physiology and Pharmacology, Federal University of Pernambuco, PE, Brazil
Rayanne Vitória Oliveira da Costa Tavares
Department of Pharmaceutical Sciences, Federal University of Pernambuco, PE, Brazil
Nataly Souza Lima
Department of Pharmaceutical Sciences, Federal University of Pernambuco, PE, Brazil
Maria Bernadete Souza Maia
Filipe Silveira Duarte
INTRODUCTION: AD is a neurodegenerative disorder characterized by a decline in cognitive function.
One of the possible mechanisms involved in the pathophysiology of AD is the oxidative stress. Unfortunately, AD
still lacks effective treatments. The phytochemical study of J. pectoralis revealed the presence of substances
with potential pharmacological activity, including flavonoids and coumarins, known by antioxidant properties. The
aim of this study was to investigate the possi-ble neuroprotective effect of HEJP against streptozotocin (STZ)induced memory impairment in rats. METHODS: All experiments were approved by the local ethics committees
(CEUA, 23076.013878/2015-11). Adult male Wistar rats were stereotaxically implanted with a guide cannu-la
directed to the lateral ventricles (i.c.v.). After three days, the animals were divided into two ex-perimental
protocols: 1) rats received vehicle (VE) or HEJP 100 mg/kg by oral route and after 1 h were i.c.v. treated with
PBS or STZ 0,1 mg (4 µL); 2) rats received VE or HEJP 100 mg/kg by oral route and after 1 h were i.c.v. treated
with PBS or STZ followed by the repeated treatment with HEJP or VE for 2 more days. The working memory
(WM), short-term (STM) and long-term memo-ries (LTM) were evaluated by the step-down inhibitory avoidance
test after receiving a scrambled foot shock. RESULTS: STZ impaired the WM (C=127±25; STZ=27±7), STM
(C=150±20; STZ+34±8) and LTM (C=142±22; STZ=34±6). The acute treatment with HEJP only protected the
STZ-induced WM impairment, without affecting the STM and LTM. In the repeated treatment, HEJP prevented the
STZ-induced memory impairment on the WM (STZ=82±29; HEJP+STZ=133±25), STM (STZ=53±22;
HEJP+STZ=151±21) and LTM (STZ=24±6; HEJP+STZ=158±14). CONCLUSION: Our results propose that HEJP
is capable of preventing the loss of memory induced by STZ, suggesting that J. pectoralis has potential
neuroprotective effect in animal model of DA. FINANCIAL SUPPORT: FACEPE.
Financial Support
FACEPE
Ethical approval
CEUA, nº 23076.013878/2015-11
359
IMPACT OF MENINGITIS CAUSED BY Cryptococcus neoformans IN WISTAR RATS IN
AZOLE'S PENETRATION IN THE BRAIN
Izabel Almeida Alves
Graziela Lock
Stela Rates
Bibiana Verlindo Araújo
Introduction: Meningitis caused by Cryptococcus neoformans is an important disease associated to high
levels of mortality, which is usually treated by Amphotericin B (ANF). Due to side effects such as nefrotoxicity,
this drug is being replaced by safer ones like fluconazole (FLU) and voriconazole (VRC). In this scenario it is
pretty important to know the drug's ability to reach the biophase, in this specific case, the brain. In the present
study we applied the microdialysis as a tool to determine the free levels reached by two drugs used in brain's
infections, during the experimental disseminative criptococosis infection in rats. To evaluate free levels of VRC
and FLU in brain of healthy and infected Wistar rats employing MD, and to establish the relationship between free
cerebral/free plasma concentrations (Ft) in both conditions. Methods: The infection was induced by the i.v.
administration of 100µL of inoculum (1-105 CFU). The animals were submitted to surgery to insert a MD probe
(CMA® 12) into the brain tissue in the region of the motor cortex (A:2.2mm; L:2.8mm; P:2.6mm). The doses was
5mg/kg i.v. of VRC or 20mg/kg i.v. of FLU. Plasma and MD concentration in the samples were determined by
LC-MS/MS and LC-UV methods. Results and Discussion: Tissue penetration of VRC was higher then FLU, in
both groups, with average Ft of 1.07 and 0.75 (p=0.03) in healthy and Ft of 2.15 and 1.02 (p=0.02) in infected,
respectively. This fact may be explained by the lipophilicity of the VRC (Log P 1.82 versus Log P 0.56) and by the
efflux transporters able to remove FLU from the brain. The increase penetration of infected tissue is due to
vascular injury produced by C. neoformans and changes in the pH of the tissue (7.4 to 5.4) causing ion trapping
(pKaVRC = 11.52; pKaFLU = 12.71). Conclusion: VRC and FLU present a good potential to be used in treatment
of brain infections caused by C. neoformans.
Financial Support
CNPq
Ethical approval
Ethics Committee in the Use of Animals UFRGS # 26605; Project name ¿Modelagem Farmacocinética-Farmacodinâmica e avaliação da penetração cerebral de
antifúngicos azólicos em animais sadios e com meningite induzida por Cryptococcus neoformans¿.
360
Cytotoxic effects of PCL (poly-ɛ-caprolactone) nanocapsules and nanospheres in
HepG2 and Vero cell lines
Izabel Cristina Trindade
Universidade Federal de Ouro Preto (UFOP)
Douglas Gualberto Sales Pereira
Giane Martins Garcia
Vanessa Carla Furtado Mosqueira
Introduction: Polymeric nanoparticulated systems show promise as active vectors due to their capacity
to release drugs, subcellular size and high intracellular uptake. Moreover, have advantage to be biocompatible and
biodegradable. However, there are few studies about its safety. The aim of this work is to assess the citotoxicity
of poly (ɛ-caprolactone) (PCL) nanoparticles on HepG2 and Vero cells, which represents liver and kidney tissues
subjected to high parenteral and oral drug exposure. Methods: Nanocapsules (NC) and nanospheres (NS) were
produced by the method of nanoprecipitation. HepG2 and Vero cells were cultivated in complete DMEM media
and maintained at 37 °C, 5% of CO2. Cells were transferred to 96-well plate and 24h later, the medium was then
replaced with 200 μl medium with NC and NS of PCL in concentrations range of 0-1000 µg/ml for 24 and 48h.
After, the medium were removed and the cells viability was assessed by MTT assay. Results and discussion:
Vero cells showed viability below 70% for all concentration and time tested, except in 1µg/ml. On the other hand,
in HepG2 cells, only PCL-NC was citotoxic at concentrations above 500 and 100 µg/ml, in 24 and 48h,
respectively. It is probable that the differences between viability profiles of these cells occurred due to the
characteristics of the tissue origin of HepG2 (human hepatocellular carcinoma) and Vero (normal monkey kidney
epithelium). The higher toxicity of NC in comparison to NS can be occurred due to the presence of
phosphatidylcholine in their composition. Conclusion: Despite PCL nanoparticles are extensively studied, the
present study showed that these particles were toxics for Vero cells, even under low concentrations. On the other
hand, its effects on HepG2 occurred at a lower rate. Although research in nanomaterials is increasing no clear
regulatory guideline on the testing/evaluation is available. Thus, in vitro toxicological studies become very
important on this area.
Financial Support
FAPEMIG, CAPES, CNPQ, UFOP, REDE NANOBIOMG
Acknowledgments
UFOP, Cipharma, REDE NANOBIOMG
361
Lycopodiella cernua L. (Lycopodiaceae) alkaloids extract as a new source of bioactive
molecules targeting Alzheimer's disease in vitro on C6 astroglial cells
Janaína Lucas de Oliveira Salomón
Hellen Knecht
Larissa Bobermin
João Paulo A. dos Santos
Carlos Alberto S. Gonçalves
Eduardo L. Konrath
Introduction: The alkaloids belonging to Lycopodiaceae species have been long investigated as potential
new biological targets, aiming multiple strategies for the treatment of Alzheimer's disease (AD). In this sense, the
in vitro evaluation of the effects of the Lycopodiella cernua alkaloids enriched fraction on a C6 astroglial lineage
cells regarding cell viability parameters (neutral red and MTT), membrane integrity (LDH), glutamate uptake and
secretion of S100B were evaluated. Materials and Methods: The aerial parts of L. cernua, were submitted to acidbase extraction and the alkaloids fraction obtained was analyzed by GC-MS to enable the identification of the
chemical profile. This enriched fraction was tested under different concentrations in a C6 astroglial lineage, during
a 24 h incubation period and different biochemical parameters were analyzed. Results and discussion: Analysis
performed by TLC allowed the visualization of two spots revealed by Dragendorff. Later, these alkaloids were
identified by GC-MS as cernuine and lycocernuine, through their fragmentation profiles, products previously
described in the literature for this species. After treatment with the alkaloids fraction on the C6 cells (25, 100 and
150 µg/mL), it was possible to verify the absence of toxic effects (MTT assay), but for neutral red assay,
cytotoxicity was verified at 150 µg/mL. No significant effects were observed on membrane integrity (LDH assay)
and over S100B secretion, while all concentrations promoted an increase in the extracellular glutamate uptake.
Such an effect may be associated to a protection of the neurons against neurotoxicity, also observed in the AD,
indicating a beneficial effect of these alkaloids to neural cells. Conclusions: The alkaloids present in L. cernua
possess anticholinesterase activity, and their effects on astroglial cells indicate that this species seems to be a
good source of molecules with multi-target potential therapeutic activity for AD.
Financial Support
CAPES and CNPq.
362
POPULATION PHARMACOKINETIC INVESTIGATION OF GEMIFLOXACIN IN PLASMA
RAT AFTER DIFFERENT DOSES
João Victor Laureano
Teresa Dalla Costa
Introduction: Gemifloxacin (GEM) is a 4th generation fluoroquinolone with a broad-spectrum of
antimicrobial activity for treatment of community-acquired respiratory tract infections used mainly against
Streptococcus pneumonia MDR. This study aimed to develop a population model (POPPK) for GEM in healthy
rats and to identify the pharmacokinetic variability (IIV) in different doses. Materials and Methods: Blood were
collected in pre-determined time intervals up to 12h. Concentration of GEM in plasma were investigated after
administration of a single 6, 20 and 40mg/kg i.v bolus dose to male Wistar rats. The samples were analyzed
using a validated HPLC/FL method. Non linear mix effects POPPK model was performed using Monolix and noncompartimental (NCA) using Phoenix®. Results and Discussion: The POPPK parameters and microconstants
were determined by fitting plasma concentration-time profiles to two-compartment model with first order
elimination process. The typical population parameters estimated were: V1 0.484 L/Kg, K10 0.277 1/h, K12 0.615
1/h and k21 0.789 1/h. The inter-individual variability was 8, 6, 14 and 14%. The parameters calculated was
clearance (CL) 0,54 L/h, volume of distribution (Vd) 3,48 L, elimination rate constant (ke) 0,151/h and half-life
(t1/2) 4,45 h. The NCA were performed to compare with the typical values determined by the POPPK modeling,
inserting the interindividual variability. For NCA the values were: CL 0,45 ± 0,10 L/h, Vd 2,90 ± 0,78L, ke 0,16 ±
0,01h and t1/2 4,35 ± 0,21h. No statistical differences between POPPK and NCA parameters determined were
observed (α = 0.05), showing that the population was adequate model to describe the data. Conclusion: A
population pharmacokinetic model of GEM was developed for healthy rats. The population pharmacokinetic
parameters of GEM estimated in the present study may be useful for studying the concentration-effect
relationship of the drug in pre-clinical scenarios.
Financial Support
FAPERGS
363
TOPICAL APPLICATION OF MICROEMULSIONS BASED ON COPAIBA OIL RESIN
ALLEVIATES DNCB-INDUCED ATOPY-LIKE DERMATITIS
JOAO WALTER DE SOUZA DA SILVEIRA
Programa de Pós-Graduação em Ciências Farmacêuticas, Universidade Estadual da Paraíba, Campina Grande, PB, Brazil
Dorian Cordeiro Lima Júnior
Departamento de Farmácia, Universidade Estadual da Paraíba, Campina Grande, PB, Brazil
Tharcia Kiara Beserra de Oliveira
Faculdade de Ciências Medicas de Campina Grande, FCM-CG, Campina Grande, PB, Brasil
Pollianna Muniz Alves
Programa de Pós-graduação em Odontologia, Universidade Estadual da Paraíba, Campina Grande, PB, Brazil
Karina Lidianne Alcântara Saraiva
Introduction: Copaiba oil-resin is extracted from the Copaiba plant, a common tree to Latin America and
West Africa. More than 20 species is found in Brazil among them the Copaifera multijuga. In popular medicine,
this oil is used by topical and oral routes for the treatment of infections and inflammations. Microemulsions are
isotropic, thermodynamically stable transparent systems of oil, water and surfactant that can improve the
solubilization of lipophilic products. Atopic dermatitis is a chronic, relapsing, and inflammatory skin disease
associated with eczematous symptoms and IgE hyperproduction. It has been recently increasing in industrialized
countries and onset is usually during early infancy and childhood, but can also occur in adulthood. This work
purposed to develop and characterize a topical microemulsion based on Copaiba oil-resin and to evaluate its antiinflammatory activity. Methods: Pseudo-ternary diagrams phase was elaborated to screening microemulsions with
copaiba oil-resin, which were characterized and then submitted to the preliminary stability test. Anti-inflammatory
activity was evaluated using DNCB-induced atopy-like dermatitis in mice. Results and Discussion: Treatment of
microemulsions containg copaiba oil-resin 1% or 5% onto DNCB-induced atopy-like skin lesions in mice
ameliorated lesion intensity scores (erythema, hemorrhage, edema, excoriation and erosion), moreover
histological analyses demonstrated decreased thickening of the epidermis. Conclusions: Microemulsions based
on copaiba oil resin alleviates DNCB-induced atopy-like dermatitis.
Financial Support
CAPES
Ethical approval
01.001.2012 (CEUA-CESED)
364
COMPUTER SIMULATIONS TO PREDICT CHANGES IN PLASMA CONCENTRATIONS
OF CARBAMAZEPINE IN DIFFERENT LOW ADHERENCE SCENARIOS
LEANDRO TASSO
Camila Cervi Pires
Introduction: Partial adherence with a prescribed or randomly assigned dose gives rise to unintended
variability in actual drug exposure in clinical practice. There are tremendous costs associated with incomplete
and/or improper drug intake. The objective of this study was to evaluate the impact of poor medication adherence
(missed, delayed and lost doses) and replacement doses on the pharmacokinetics of carbamazepine (CBZ).
Methods: Concentration-time profiles in different scenarios were simulated on the basis of pharmacokinetic
parameters of CBZ in regimens of 200 and 400 mg three times daily (TID) and 400 mg every 12h (BID).
Simulations in Microsoft Excel® software were performed. Results and Discussion: The concentrations were
below the therapeutic range (TR) independent on the number of missed doses, except in the 400 mg TID regimen.
Doses replacements resulted in concentrations higher than the TR only at the highest dose (400 mg). The delay
of up to 6 hours at a dose of 200 mg TID regimen did not affect plasma levels of CBZ. The 400 mg TID regimen,
the interval of only 2 hours between administration delayed dose and the administration of the usual dose
exceeded the TR. In 400 mg BID regimen, any delay simulated up to 8 hours did not commit the plasma levels of
CBZ. Conclusion: Simulations based on the pharmacokinetics of CBZ in Microsoft Excel is a useful tool to
investigate the impact of poor medication adherence and provide information about replacement doses.
365
CHOLECALFICEROL MODULATES LIPOPOLYSACCHARIDE-INDUCED IMMUNE
RESPONSE IN RAW 267.4 MACROPHAGES
Leonardo Mendes Bella
Leonardo Mendes Bella
São Paulo University
Thatyane de Castro Quirino
Fernando Henrique Galvão Tessaro
Thaís Soprani Ayala
Eduardo Lima Nolasco
Carolina Bassoli de Azevedo
Federal University of São Paulo
Joilson de Oliveira Martins
Introduction Macrophages are key cells in infectious and inflammatory processes. The magnitude of
these phenomena can be regulated by modulation of cytokine release. Immunomodulatory effect of vitamin D has
been assigned by the activated form, as known as calcitriol. However, the cholecalciferol, an inactive form of
vitamin D, activity is not described as an immunomodulatory agent. Material and Methods Murine RAW 264.7
macrophages were cultured in hormone-free medium (DMEM) supplemented with penicillin (40 U/mL) and
streptomycin (50 µg/mL). Cells (2x106) were plated in 12-well plates for 4h and 24 h at 37°C with 5% CO2
atmosphere and divided in four groups (n= 6, duplicate): control; LPS (100ng/mL); vitamin D (100nM/mL); LPS +
vitamin D (100nM/mL). IL 1β, IL-6 and IL-10 were measured by ELISA; NO by Griess and cell viability by MTT
assay. Results Cell viability not showed differences between four groups. When compared IL1β, IL-6, IL-10 and
NO released, there were no differences between cells threated with cholecalciferol and control group after 4h and
24h. However, cells stimulated with LPS released more IL 1β, IL-6, IL-10 and NO than control group after 4h and
24h of stimulation. Interestingly, cells stimulated with LPS and threated with cholecalciferol released less IL 1β,
IL-6 and NO than cells stimulated with LPS after 4h (IL1β: 227.4+15.37 vs 170.4+37.11, p<0,01; IL-6:
388.8+32.47 vs 313.9+14.39, p<0,001) and 24h (IL1β: 129.2+24.39 vs 65.52+15.79, p<0,001; IL6: 985.8+38.55
vs 814.5+53.58, p<0,001 and NO: 47.55+6.58 vs 28.67+3.69). When compared IL-10 released, there were no
differences between cells stimulated with LPS and threated with cholecalciferol and cells stimulated with LPS
after 4h and 24h. Conclusion These results suggest that cholecalciferol might modulate the release of IL 1β, IL6
and NO by RAW 267.4 cell.
Financial Support
Pró-reitoria de pesquisa; FAPESP;CNPq
Ethical approval
CEUA FCF/USP 464
366
OXIDATIVE STRESS PARAMETERS IN PATIENTS WITH VIRAL HEPATITIS C IN
TREATMENT WITH RIBAVIRIN AND PEGYLATED ALPHA INFERFERON
Lyana Feijoó Berro
Juliana Mezzomo
Patrícia Martinez Oliveira
Jacqueline da Costa Escobar Piccoli,
Raqueli Bittencourt
Secretaria Municipal de Saúde de Uruguaiana, RS, Brazil.
Mariana Ziolkowski
Secretaria Municipal de Saúde de Uruguaiana, RS, Brazil.
Vinicius Tejada Nunes
Vanusa Manfredini
Introduction: Viral hepatitis performs a major public health problem, and the hepatitis C virus (HCV), the
most prevalent. Recent studies indicate that patients with viral hepatitis C may develop serious liver damage and
impairment of oxidative phosphorylation processes. It is believed that damage to the liver, mediated cytotoxicity
by different viral products, and oxidative stress have been associated with the etiology and progression of chronic
hepatitis C. Other studies also show that patients with hepatitis C treated with pegylated interferon alpha and
ribavirin have greater oxidative damage to biomolecules such as proteins and lipids. However, little is known
about the effects of these drugs in the long run. Therefore, this study aimed to evaluate the parameters of
oxidative stress in patients with viral hepatitis C during treatment with ribavirin and pegylated alpha inferferon.
Material and Methods: For this study 43 patients with viral hepatitis C (33 men and 10 women) were recruited
near to the reference service in Uruguaiana city, RS. The patients were divided into 4 groups: group 1 patients
diagnosed with viral hepatitis C; group 2: patients in early treatment; group 3: patients in the middle of treatment
and group 4: patients at the end of the treatment. After the free and informed consent term was signed, venous
blood was collected and placed in EDTA tube for further analysis. Lipid peroxidation and protein carbonyls
parameters were determined using classical methods. Results and Discussion: The time of diagnosis of TBARS
levels are significantly elevated (p <0.05) compared to the other groups and along the combined treatment of
ribavirin and pegylated alpha inferferon TBARS levels tend to decrease levels. Nevertheless the amount of
carbonylated protein increases significantly (p <0.05) during treatment. Conclusions: Thus, it is suggested that
there is oxidative damage to biomolecules along the antiviral therapy in patients with viral hepatitis C.
Financial Support
Ethical approval
Approved by the Ethics Research Committee (CEP) of the Federal University of Pampa, CEP, Uruguaiana, Rio Grande do Sul, Brazil (001/2012).
367
POPULATION PHARMACOKINETIC MODELING OF GLIMEPIRIDE IN HEALTHY AND
TYPE 2 DIABETIC RATS
Maiara Cássia Pigatto
Pharmaceutical Sciences Graduate Program, College of Pharmacy, Federal University of Rio Grande do Sul, Porto Alegre, Brazil
Jaqueline Schneider Izolan Fabricio
Andressa Braga
Centro Bioanalítico de Medicamentos, College of Pharmacy, Federal University of Rio Grande do Sul, Porto Alegre, Brazil
Teresa Dalla Costa
Introduction: Diabetes may affect the pharmacokinetics of different drugs by changing absorption,
distribution, biotransformation and excretion processes. In this way, the goal of this study was to develop a
population pharmacokinetic (popPK) model of glimepiride and to investigate the influence of type 2 diabetes (T2D)
on the popPK of glimepiride in Wistar rats. Methods: The T2D model in male Wistar rats was obtained by
administration of nicotinamide (100 mg/kg, i.p.) and streptozotocin (65 mg/kg, i.v.). Healthy (n = 11) and diabetic
(n = 9) rats received glimepiride 5 mg/kg i.v. bolus. Blood samples were taken from the tail vein up to 6 h and
quantified by HPLC-UV. The non-compartmental analysis (NCA) was done using the Phoenix® software. The
popPK modeling was conducted using MONOLIX v.4.3.3 software. The effect of categorical covariate diabetes on
glimepiride PK parameters was investigated and the final model was obtained based on Akaike's information
criterion (AIC), improvement of the precision of the parameter estimates, visual assessment of goodness-of-fit
plots, and VPCs. Results and Discussion: PK parameters determined by NCA showed t1/2 of 1.77 ± 1.23 and
2.70 ± 1.99 h, AUC0∞ of 20.49 ± 5.26 and 46.54 ± 16.35 μg.h/mL, and CLtot of 0.079 ± 0.02 and 0.036 ± 0.01
L/h for healthy and diabetic rats, respectively. Statistical difference was observed for CLtot and AUC0∞ (p <
0.05), indicating a reduction of glimepiride elimination in diabetic rats. A 2-compartment structural model with firstorder elimination was the most adequate model to describe simultaneously the glimepiride plasma concentrationtime profiles derived from healthy and diabetic rats. The T2D was an influential covariate on the parameter k10 (p
< 1.0-10, Wald test). Conclusions: The popPK model described that T2D strongly influence the interindividual
variability in the PK of glimepiride and the model can be useful to predict the changes in exposure due to disease.
Financial Support
CNPq/Brazil
Acknowledgments
CAPES Foundation/Brazil for the scholarships
Ethical approval
CEUA/UFRGS (#27892)
368
EVALUATION OF Philodryas patagoniensis VENOM TOXICITY IN Artemia salina AND
HUMAN LEUKOCYTE MODELS
Márcio Tavares Costa
Biochemistry Graduate Program, University of Pampa, Uruguaiana, RS, Brazil
Aline Flores da Silva
Faculty of Natural Science, University of Pampa, Uruguaiana, RS, Brazil
Luciana de Mello Viana
Faculty of Physiotherapy, University of Pampa, Uruguaiana, RS, Brazil
Andréia Caroline Fernandes Salgueiro
Hemerson Silva da Rosa
Vanderlei Folmer
In 1918 started researches with opisthoglyphous snakes and their venom in Brazil. Among rear-fanged
snakes, there is genus Philodryas of Dipsadidae family. In Brazilian Pampa Biome, the specie that stands out by
abundance is P. patagoniensis. Its venom produced by Duvernoy gland is target of researches in several areas.
Therefore, the aim of this study is to evaluate the P. patagoniensis venom toxicity in nauplii of Artemia salina and
in human leukocyte, and establishing the LD50, as well as correlate these data. For this, snakes captured in
Uruguaiana/RS had their poison extracted for analyses. Concentration of venom proteins were measured by
Bradford method. Evaluation of toxicity in nauplii was performed using a microtiter 96 well plates. Nauplii were
individually arranged and exposed to venom in different concentrations. After 24 hours, the brine shrimp mortality
was measured. For LD50 determination was used logarithmic regression curve. To cell viability bioassays,
mononuclear leukocytes were exposed to snake venom in different concentrations, and after incubation time,
were evaluated the cell viability by trypan blue method. Statistical analysis were performed by one way ANOVA
and Bonferroni post hoc test. Value of p ≤ 0.05 were used to determine statistical significance. Venom protein
determined was 115.7 mg/mL. Tests in A. salina indicated that best doses to be worked were 1:100, 1:200, 1:500,
1:1000, and 1:2000. LD50 determined in this model is 461 µg/mL. Cell viability tests illustrated damage caused by
venom in two dilutions doses: 1:100 and 1:200. No significant differences were observed in sublethal doses.
These data show medium toxicity venom in A. salina and direct relationship between nauplii mortality and cellular
viability in toxicological evaluation of P. patagoniensis venom. Here, we emphasize these models as an
alternative to testing ophidians toxins.
Financial Support
CNPq
Ethical approval
Ethics Committee on Animal Use through the protocol 031/2014. Chico Mendes Institute for Biodiversity Conservation registered by the number 4569 1-1
process.
369
CYTOTOXIC EVALUTION AND APOPTOSIS INDUCTION OF IMIDAZACRIDINE
DERIVATIVE LPSF/AC05 IN LYMPHOMA AND LEUKEMIA CELLS
Valécia de Cassia Mendonça da Costa
Ivan da Rocha Pitta
Moacyr de Jesus Barreto de Melo Rêgo
Introduction: Cancer has become a major cause of morbidity and mortality worldwide. Most currently
available treatments are associated to severe adverse effects and drug resistance, which stimulates the
development of novel selective antineoplastic drugs. The present abstract describes the citotoxicity activity and
apoptosis induction of the imidazacridine derivative (LPSF/AC05). Methods: Cytotoxicity assays were performed
in Peripheral Blood Mononuclear Cells (PBMCs) and in six lymphoma (Raji and Toledo) and leukemia (Jurkat, HL60, K-562 and CCRF-CEM) cell lines using MTT method. Cells were treated with LPSF/AC05 (1 to 75 μM). Nontreated cells were used as negative control, amsacrine was used as positive control and DMSO (0.1%) as vehicle
control. Cell apoptosis assays were performed by flow cytometry using Apo-BrdU Apoptosis Detection Kit (BD,
USA) according to manufacturer's instructions. Results and discussion: Cytotoxicity assays performed in
neoplastic cells showed best results of LPSF/AC05 in Toledo, Jurkat, Raji and CCRF-CEM (IC50=27.18, 31.04,
33.36 and 54.94 μM, respectively). LPSF/AC05 showed no toxicity against HL-60, K-562 and PBMCs (IC50>75
μM). Amsacrine showed IC50 value against PBMCs (IC50=3.7 μM). Anticancer properties of acridinas
derivatives have been related to their planar aromatic structure that enables interaction with DNA. LPSF/AC05
increased significantly (p<0.05) the percentage of apoptotic cells in all cell lines tested when compared to nontreated cells (Toledo, p=0.015; Jurkat, p=0.004; Raji, p=0.004; CCRF-CEM, p=0.0003) and also when compared
to amsacrine (Toledo, p=0.045; Jurkat, p=0.003; Raji, p=0.006; CCRF-CEM, p=0.0008), indicating that apoptosis
is probably a major cell death mechanism induced by LPSF/AC05. Conclusions: LPSF/AC05 showed selectivity
for PBMCs, cytotoxic effects in tumoral cells in vitro and apoptosis induction. Therefore, the compound shows
potential as a new candidate for cancer therapy.
Financial Support
CNPq, FACEPE, INCT_if
Acknowledgments
PPGIT, UFPE
Ethical approval
CEP: 11006 CCS/UFPE
370
LYSINE-BASED SURFACTANTS WITH LOW SKIN IRRITATION EFFECT ASSESSED
WITH CULTURED KERATINOCYTES AND A HUMAN EPIDERMIS EQUIVALENT MODEL
Montserrat Mitjans
Introduction. Surfactants are among the most common and versatile components of pharmaceutical and
cosmetic formulations. The search of surfactants with non-irritant effects on the skin is of great interest. In this
sense, anionic dyacyl lysine-based surfactants prepared as mimics of lecithins and with different counterions
constituted a promising strategy. Methods. The potential skin irritation induced by lysine-based surfactants with
Potassium, Lithium, Sodium, or Lysine as counterions has been studied in vitro using cell culture of HaCaT
keratinocytes and Episkin as a human epidermis equivalent model as alternatives to the in vivo Draize test.
HaCaT cells were incubated for 24 h under 5% CO2 at 37ºC in presence of the surfactants (7.8-500 µg/ml). Each
concentration was tested in triplicate and control cells were exposed to medium. Surfactants were added to the
Episkin model at concentration of 1 mg/ml and incubated for 24 h. The viability of the cells after the incubation
period with the surfactants was assessed in the two models by the MTT reduction assay. Results are compared
to sodium dodecyl sulphate (SDS) as a known skin irritant surfactant. Results and Discussion. The
concentration-response curves obtained from the MTT assay in HaCaT cells were obtained for each surfactant
and the concentrations that inhibited 50% of viability (IC50) were determined. The lysine-based surfactants
presented IC50 many times higher than SDS. The mean viability in the Episkin model was from 84 to 100% for
the lysine-based surfactants and 18% for the SDS. Conclusions. According to the results obtained we concluded
that the lysine-derived surfactants are less irritants than the commercial SDS and could be classified as low
irritants. For this reason they may be a good alternative for use in topical pharmaceutical and cosmetic
formulations.
Financial Support
Ministerio de Economía y Competitividad - Spain and European Union (FEDER) (Project MAT2012-38047-C02-01)
371
INFLUENCE OF INHALATION EXPOSURE TO TOLUENE ON THE PHARMACOKINETICS
OF PRAVASTATIN AS AN IN VIVO PROBE DRUG OF OATP1B1 IN RATS
Mariana Mauro
Departamento de Princípios Ativos Naturais e Toxicologia, Faculdade de Ciências Farmacêuticas, Universidade Estadual Paulista,
Araraquara, SP, Brazil
José Salvador Lepera
Bruno Borsari
Jorge Manuel Vieira Capela
Departamento de Físico-Química, Instituto de Química, Universidade Estadual Paulista, Araraquara, SP, Brazil
Natália Valadares de Moraes
Introduction: Occupational exposure to chemicals are potentially able to modify the kinetic disposition
and drug metabolism. However, studies showing the impact of occupational exposure to chemicals in the activity
of drug transporters are scarce. This study evaluates the influence of inhalation exposure to toluene on the in vivo
activity of the organic anion transporter polypeptide 1B1 (OATP1B1) in rats, using pravastatin as a probe drug.
Clinical and in vitro data have demonstrated that alterations in the kinetics of pravastatin are observed when the
OATP1B1 activity is inhibited. Methods: Male Wistar rats exposed to filtered air (control group) or to toluene (85
mg/m3) in a nose-only inhalation chamber during 6h/day, 5 days/week, 4 weeks received a single oral dose of 20
mg/kg pravastatin. Plasma concentrations of pravastatin were analysed by high-performance
liquid
chromatography coupled to mass spectrometry detection (LC-MS). The LC-MS method for pravastatin analysis in
rat plasma was validated by evaluation of matrix effect, linearity, lower limit of quantification (LOQ), within-run and
between-run precision and accuracy and stability studies. The method was successfully applied in the
investigation of pravastatin single dose pharmacokinetics in rats. Results and Discussion: The areas under the
plasma concentration versus time curves (AUC0∞) were calculated by the Gauss-Laguerre Quadrature and total
apparent clearance (CLT/F) was calculated from the equation: CLT/F=dose/AUC0∞. Non-exposed animals
showed AUC0∞ of 726 ng¿h/mL and CL/F of 49.1 L/h.kg. There was no statistical difference in the AUC0∞
when toluene-exposed rats (AUC: 681.8 ng¿h/mL) were compared to control group. Conclusion: The exposure to
85 mg/m3 toluene by inhalation (6h/day, 5 days/week, 4 weeks) in rats did not alter the activity of OATP1B1
evaluated using pravastatin as a probe drug.
Financial Support
CNPq, CAPES
Ethical approval
Ethics Committee for Use of Animals of School of Pharmaceutical Sciences, UNESP (Protocol number: CEUA/FCF/CAr nº 21/2014)
372
ANTINOCICEPTIVE AND ANTI-INFLAMMATORY ACTIVITY OF Aspidosperma pyrifolium
Mart - APOCYNACEAE
Pablo Rayff da Silva
Renaly Ivyna de Araújo Rêgo
Raiff dos Santos Dantas
Hilton César Pereira de Albuquerque
Jhonatta Alexandre Brito Dias
Maciel Araujo Oliveira
Ingrid Laiz de Oliveira Cabral
Ana Claudia Dantas de Medeiros
Vanda Lúcia dos Santos
Pain is a sensory experience that occurs over the inflammatory process, caused by cell or tissue
damage. The folk use of medicinal plants for therapeutic purposes such as analgesic and anti-inflammatory
functions is well known and provided some useful drugs. This research aimed to investigate the antinociceptive
and anti-inflammatory activity of the ethanolic extract from the peel of Aspidosperma pyrifolium, popularly known
as Pereiro, registered in voucher specimen under the number 20104. Males and Females Swiss mice were used
for in vivo tests. Experimental protocols were approved by the Ethics Committee on Animal Use of Higher
Education and Development Center (No.0020/16102012). Models of abdominal writhes induced by acetic acid and
Formalin Test were used to determine the antinociceptive activity, as well as the Paw Edema Test and Peritonits
induced by carrageenan were applied for the anti-inflamatory activity. The data were analyzed using the analysis
of variance followed by the Test of Dunnet through the GraphPadPrism 5.0. software. The extract was used in
doses of 62.5, 125 or 250 mg/g(via oral). Analgesic activity's results shows that the extract cause 100% inhibition
of abdominal writhing, and a significant inhibition of the nociception induced by formalin in 83.3, 97.2 and 65.7%
for each dose, respectively. For inflammatory activity, the extract inhibited in 45.0, 60.6 and 54.8% the Paw
Edema, and the leukocyte migration to the inflammatory site were inhibited in 29.0, 49.5 and 59.1%, for the dose
of 62.5, 125 or 250mg /g, respectively, when compared to the negative control. These activities are related to the
presence of indole alkaloids (vincadifformine, pirifolidinmine, aspidospermine) representative of Aspidosperma
genre identified by chromatography. It is concluded that were antinociceptive and anti-inflammatory activities for
the peel of Aspidosperma pyrifolium extract, constituting a promising source for the development of new drugs.
Financial Support
CNPq and UEPB
Ethical approval
No. 0020/16102012
373
ANALYSIS OF COCAINE/CRACK BIOMARKERS IN MECONIUM BY LC-MS
Pamela C Lukasewicz Ferreira
Felipe B D'Avila
Fernanda R Salazar
Andrea G Pereira
Maira K Santos
Flavio Pechansky
Hospital de Clinicas de Porto Alegre
Renata P Limberger
Pedro E Froehlich
Introduction Fetal exposure to illicit drugs is a worldwide problem, since many addicted women do not
stop using it during pregnancy and it may cause many adverse effects to the newborns. Cocaine consumed in
powder (snorted or injected) or smoked (crack cocaine) is harmful for the baby and its side effects are not
completely known. Meconium, the first stool of a newborn, is a matrix usually discarded, that may contain
amounts of substances consumed in the last two trimesters of pregnancy. Analyzing this biological matrix it is
possible to detect the unaltered molecule of cocaine (COC) or its major metabolite benzoylecgonine (BZE) as well
as its pyrolytic products from crack use anhydroecgonine methyl ester (AEME) and anhydroecgonine (AEC).
Method A liquid chromatography mass spectrometry (LC-MS) method was validated for meconium samples after
solvent extraction followed by direct injection. Linearity covered a concentration range of 15 to 500 ng/mg with a
lower limit of quantification (LLOQ) of 15 ng/mg for all analytes. Results Validation was performed following
mainly the FDA bioanalytical method validation guideline and complemented by SWGTOX guide. Selectivity, limit
of detection (LOD), limit of quantification (LLOQ), linearity, precision, accuracy, matrix effect and stability were
accessed. Discussion Detection of illicit substances usage can be crucial for the newborn, since knowing that
can help provide adequate medical care as fast as possible. The developed method proved to be simple and fast,
and was applied to 17 real meconium samples. On eleven was detect COC and BZE suggesting the consumption
in powdered form, while AEME and AEC were detect in three and two samples, respectively. Conclusion Simple
and fast cleaning procedure using small quantities of solvents was applied successfully for the complex biological
matrix meconium. An LC-MS method was fully validated allowing differentiating the form of consumption
depending on the analyte found.
Financial Support
The authors want to thank
CNPq and CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior) for the financial s upport
Acknowledgments
Dr. Gabrielle Bochese, Dr. Flávio Pechansky (HCPA), Dr. Bruno Todeschini and Dr. Adriano Maldaner (Federal Police)
Ethical approval
The protocol of the study was formally accepted by the Ethics Committee of Hospital de Clínicas de Porto Alegre (110329), Hospital Materno Infantil Presidente
Vargas (25/11) and Santa Casa de Caridade de Bagé, Rio Grande do Sul ¿ Brazil. Informed consent was given for the subjects after explanations about the
possible harms and purposes of the research, assuring that all data are confidential
374
ANTIOXIDANT ACTIVITY AFTER SUBACUTE TREATMENT WITH Olea europaea L. IN
LIVER AND KIDNEY OF wistar RATS
Patricia Romualdo de Jesus
Camille Gaube Guex
Fernanda Ziegler Reginato
Edimara Machado Lopes
Pedro Piovesan Lago
Kássia Caroline Figueredo
Andreia Regina Haas da Silva
Gilberti Helena Hübscher Lopes
Liliane de Freitas Bauermann
Introduction: The production of reactive oxygen species (ROS) is necessary for biological functions of
the body. However, when produced in excess can cause a damaging process to the cells. The body has an
antioxidant defense system for balance of ROS and preventing oxidative stress. Several plants have antioxidant
activity, including the Olea europaea L. species, known as olive. The aim of this study was to assess the lipid
peroxidation by the thiobarbituric acid reactive substances (TBARS) method and antioxidant activity by vitamin C
levels after the treatment with tincture of olive leaves (TOL) for 28 days. Methods: TOL was obtained from the
company FLORIEN - Flores e Ervas Com. Farm. Ltda - EPP. Wistar rats (weighing 150-250 g) were obtained
from animal house of Universidade Federal de Santa Maria (UFSM). The study was approved by Comitê de Ética
no Uso de Animais from UFSM under the number 121/2014. Following the OECD 423 protocol, animals were
divided into 4 groups: Group I. Control - Ethanol 51% (tincture vehicle -10 mL/kg) Group II. TOL (100 mg/kg)
Group III. TOL (200 mg/kg) and group IV. TOL (400 mg/kg) and treated for 28 days by oral gavage. On the 29th
day, animals were euthanized by cardiac puncture. After, kidney and liver were collected and homogenized for
analysis by the TBARS method and vitamin C. Data were analyzed by one-way ANOVA, followed by Tukey.
Differences between groups were significant when p<0.05. Results and Discussion: Treatment with TOL showed
a significant difference in vitamin C levels between the control and treated groups (p<0.05). TBARS method
quantifies the levels of malondialdehyde, end product of lipid peroxidation, and vitamin C acts to increase the
absorption of iron and inactivating ROS, being important markers of oxidative damage and antioxidant activity.
Conclusions: The results indicated an increase in antioxidant defense. However, further studies are needed to
elucidate the antioxidant activity of the plant.
Financial Support
CAPES
Acknowledgments
UFSM and Bioclin
Ethical approval
The study was approved by Comitê de Ética no Uso de Animais from UFSM under the number 121/2014.
375
INFLUENCE OF ALCOHOL PRESENT IN ESPECÍFICO-PESSÔA TINCTURE ON RENAL
AND LIVER FUNCTION IN RATS.
PRISCILA DA CAZ
UNIOESTE - University of Western Paraná.
PRISCILA DA CAZ
FERNANDA COLERAUS SILVA
ANA MARIA ITINOSE
CEATOX - Assistence Center in Toxicology, Hospital University of Western Paraná.
CARLA BRUGIN MAREK
SUÉLYN KOERICH
FABIANA SARI FERREIRA
INTRODUCTION: Específico-Pessôa is a hidroalcoholic phytotherapic tincture that has been used for
more than 30 years as additional therapy for snake bites in Brazil. The presence of the alcohol, which is important
to extract bioactive substances from plants, can interfere in action of bioactive components presents in herbal
medicines. The study investigated the possible changes with the alcohol presence on the lipid profile and renal
function of rats as well as their pharmacodynamics effects. METHODS: Three groups of male Wistar rats, with
five animals randomly allocated in each, were treated orally once a day for 10, 15 and 30 days: 1.Control group
(water); 2.Alcohol diluted in water (2.77 mg?kg?1, referring to this dose of alcohol in the tincture) and 3.Tincture
(0.25mL?kg?1, that was diluted in 14 ml of water), a total of 9 groups in experiment. At the end of the study the
animals were anesthetized for blood collection and euthanized. The blood colection for biochemical determinations
(cholesterol, triglyceride, uric acid and creatinine) was obtained by intracardiac puncture. For the statistical
analysis Dunnett test was used and the statistical significance was accepted at p < 0.05. RESULTS AND
DISCUSSION: Microscopy analysis showed renal damage. Uric acid levels have been changed and the
creatinine increased in alcohol and tincture groups after 15 and 30 days of experiment. In liver, was observed
polymorphonuclear cells and hydropic multifocal degeneration with sligh lymphohistiocytic infiltration. The levels
of cholesterol increased 25% in alcohol group (p < 0.05) and 26% in tincture group (p < 0.05) both with 10 days of
treatment. The triglyceride levels decreased in tincture group 36% (p < 0.05) and 30% (p < 0.01) after 15 and 30
days of experiment respectively. CONCLUSION: The effect of the alcohol presence in Específico-Pessôa
demonstrated serious biochemical alterations, with renal function in lipid profile changed as well as liver damage.
Financial Support
HIGHER EDUCATION PERSONNEL IMPROVEMENT COORDINATION (CAPES).
Acknowledgments
I THANK MY COLLEAGUES IN THE LABORATORY OF TOXICOLOGY AT UNIOESTE, THAT HELPED ME AT EXECUTION OF THIS S TUDY, ESPECIALLY THE
TEACHERS ANA MARIA ITINOSE, FERNANDA COLERAUS SILVA AND CARLA BRUGIN MAREK.
Ethical approval
ETHICS COMMITTEE IN THE USED OF ANIMALS (CEUA) FROM THE STATE UNIVERSITY OF WEST PARANÁ (NUMBER 067/11)
376
POPULATIONAL PHARMACOKINETIC MODELING OF PLASMA AND TISSUE
CONCENTRATION OF TOBRAMYCIN IN HEALTHY AND Pseudomonas aeruginosa
BIOFILM-FORMING INFECTED WISTAR RATS
Priscila Martini Bernardi
Pharmaceutical Sciences Graduate program, Federal University of Rio Grande do Sul, Porto Alegre, RS, Brazil.
Fabiano Barreto
Laboratório Nacional Agropecuário do Rio Grande do Sul (LANAGRO/RS), Porto Alegre, RS, Brazil.
Teresa Dalla Costa
Introduction: Biofilms reduce tobramycin (TOB) bactericidal activity in vitro, but the influence of this
infection on the drug pharmacokinetics (PK) is unknown. Objective: To investigate the impact of P. aeruginosa
biofilm-infection on TOB's lung penetration and to establish a population PK model (popPK) to describe lung and
plasma data in healthy and infected rats. Methods: Biofilm-forming P. aeruginosa (PA14) was intratracheally
inoculated (109 CFU/mL) to male Wistar rats. Seven days after, anesthetized and ventilated infected rats (n = 5)
received TOB 10 mg/kg i.v. bolus dose. Control healthy group (n = 5) received the same dose. Blood and lung
microdialysate (CMA/20 probe) samples were collected up to 12 h post-dose. TOB binding to healthy and infected
rats' plasma protein was determined by microdialysis. Samples were quantified by LC-MS/MS. Noncompartmental analysis (NCA) and the popPK modeling were done using Phoenix® and Monolix®, respectively.
Results and Discussion: TOB protein binding (11.3 ± 1.9%) was independent of plasma origin. Infection did not
affect plasma PK. Pneumonia reduced TOB lung penetration factor (AUCfree,lung/AUCplasma¿fu) from 0.94 to
0.25. A two-compartment popPK model described simultaneously TOB total plasma and free lung concentrations
in both groups. Plasma parameters determined by popPK (CL: 0.02 L/h; ASC0∞: 150 µg·h/mL and Vdss: 0.13 L)
were similar to those determined by NCA (CL: 0.021 L/h; ASC0∞: 136 µg·h/mL and Vdss: 0.14 L). The
categorical covariate "infection" allowed to describe the increase in the volume of the peripheral compartment
(from 0.084 L to 0.396 L), and in the reduction on the elimination rated constant from the central compartment
(from 0.568 h-1 to 0.397 h-1). Conclusion: Total plasma levels overstimate active TOB lung concentrations in P.
aeruginosa biofilm-forming pneumonia. The popPK model allows predicting TOB free concentrations in pnemonic
lung using total plasma data.
Financial Support
CNPq/Brazil, CAPES/Brazil.
Ethical approval
Approved by UFRGS CEUA (27847)
377
INFLUENCE OF DULOXETINE AND AMITRIPTYLINE IN THE BIOLOGICAL EFFECT OF
PREGABALIN FOR CHRONIC PAIN TREATMENT IN ANIMALS
Rafaela Figueiredo Rodrigues
RODRIGUES, R.F
Pharmaceutical Sciences Graduate Program, Federal University of Alfenas, Alfenas, MG, Brazil; Alfenas, MG, Brazil
SANTOS, R. S
Multicentre Program in Physiological Sciences - Graduate Program, Federal University of Alfenas, Alfenas, MG, Brazil
GALDINO, G. S.
Multicentre Program in Physiological Sciences - Graduate Program, Federal University of Alfenas, Alfenas, MG, Brazil
CARDOSO, M. H. M.
BARROS, C.M.
Santa Casa de Alfenas, Alfenas, MG, Brazil
BORALLI, V.B
Pain is an unpleasant sensory and emotional experience that occurs in varying degrees of intensity. The
pain can be classified as acute and chronic, including neuropathic pain. Neuropathic pain is more complex mainly
due to its resistance to common analgesics. The Brazilian Ministry of Health advocated for treating this type of
pain in humans with pregabalin(PREG), duloxetine(DULOX) and amitriptyline(AMITRIP). The main purpose of this
work was to evaluate if medicaments association could reduce nociceptive perception in rats. Wistar male rats
(220-250g, n=12 for each group), approved by Ethical Comite (UNIFAL-MG) - number 670/2015, with neuropathic
pain induced by chronic constriction injury of sciatic nerve; pain assessment was done by the von Frey hair test.
The biological effect tests were performed in six groups: PREG10mg/kg; PREG10mg/kg+AMITRIP1mg/kg;
PREG10mg/kg+DULOX30mg/kg; AMITRIP1mg/kg; DULOX30mg/kg; vehicle (ultrapure water) (VEH). The data
was evaluated using ANOVA. The results showed that PREG, PREG+AMITRIP and PREG+DULOX treated
groups showed better analgesic effect compared to VEH group, specially at 2:15 and 4h for PREG; 2:15, 4 and
8h for PREG+AMITRIP; 4 and 8h for PREG +DULOX; after substances administration. Otherwise, AMITRIP and
DULOX groups showed no antinociceptive effect. Compared to VEH, the PREG, PREG+AMITRIP and
PREG+DULOX groups had increased nociceptive threshold in 10.4; 12.5 and 19.4 times, respectively, after
administration of the substances. Besides, PREG+AMITRIP and PREG+DULOX groups had prolonged analgesic
effect (maximum effect 8 hours after administration) better than the PREG group (maximum effect 4 hours after
administration). Additionally, all false-operated group (sham) presented hypoalgesia after substances
administration when compared to control group. Therefore, coadministration showed benefits against the use of
pregabalin alone for the relief of nociceptive effect.
Financial Support
CAPES
Ethical approval
Ethical Comite (UNIFAL-MG) by the number 670/2015
378
PHARMACOLOGICAL ACTIVITY OF Spondias purpurea Mart - ANACARDIACEAE
Renaly Ivyna de Araújo Rêgo
Hilton César Pereira de Albuquerque
Ingrid Laiz de Oliveira Cabral
Jhonatta Alexandre Brito Dias
Maciel Araújo Oliveira
Pablo Rayff da Silva
Raiff dos Santos Dantas
Ana Claudia Dantas de Medeiros
Vanda Lúcia dos Santos
The inflamatory process is an organism reaction towards an infection or tissue damage, triggered by
vasodilation and hyperemia, that are caused by chemical mediators (histamine, bradykinin and prostaglandin). The
usage of derived substances from natural products to combat the inflammatory process has been gaining
emphasis and currently many products has value in the therapeutic use. The objective of this work was to
evaluate the toxicological, anti-inflammatory, antinociceptive and antiulcer activity of ethanolic extract (EEtOHSp), fractions of ethyl acetate (FaAce-Sp) and dichloromethane (FaDic-Sp) of the S. purpurea leaves (Seriguela).
Males and females Swiss mice were used. The experimental protocols were approved by the Ethics Committee
on Animal Use of Higher Education and Development Center (n° 0020/16102012). For the toxicological evaluation,
the test of acute toxicity was made, with a dosage of 2g/kg. For the analysis of pharmacological activities were
performed the abdominal contortion tests induced by acetic acid, peritonitis induced by carrageenan and the ulcer
by HCl/ethanol. The results shows the low toxicity of the extract, since that there was no death, no physiological
changes (weight and water consume or food) neither macroscopic changes of the animals organs when compared
to the control group. The antinociceptive activity results shows the inhibition of 72,1; 88,4 and 27,5% to the
EEtOH-Sp, FaDic-Sp and FaAce-Sp, respectively. In the anti-inflammatory activity the leukocyte migration
inhibition to the inflammatory site was 22,7; 32,0 and 44,6% respectively. In the antiulcer activity the inhibition
was 71,1 and 77,2% to EEtOH-Sp and FaDic-Sp, respectively, corroborating with the structure-activity
relationship of the phytochemical compounds of this species. Therefore, these results demonstrate that the
extract and fractions of the S. purpurea are promising sources for the production of a new herbal medicine.
Financial Support
CNPq and UEPB.
Ethical approval
Ethics Committee on Animal Use of Higher Education and Development Center (n° 0020/16102012).
379
INTEGRATED TESTING STRATEGIES FOR SKIN SENSITIZATION EVALUATION OF
HAIR DYES COMPONENTS
Renato Ivan de Ávila
PhD student, Faculty of Pharmacy, Federal University of Goiás, Goiânia, GO, Brazil
Gabriel Campos Teixeira
Faculty of Pharmacy, Federal University of Goiás, Goiânia, GO, Brazil
Thaisângela Rodrigues Lopes e Silva Gomes
Danillo Fabrini Maciel Costa Veloso
Eliana Martins Lima
Marize Campos Valadares
INTRODUCTION Skin sensitization (SS) evaluation has historically been relied on animal models, in
particular the murine local lymph node assay (LLNA). Although SS is a complex process, its key biological events
have recently been demonstrated in an adverse outcome pathway, supporting the development of non-animal
assessment strategies. Given the above, this study performed integrated testing strategies (ITS) involving three
key-events of SS for hazard evaluation of hair dyes ingredients (HDIs). METHODS Ten HDIs, being nine
sensitizers for humans, were tested for the following assays: Key-event (1): according to OECD 442C (2015),
direct peptide reactivity assay (DPRA) was carried out by incubation of HDIs with synthetic cysteine- or lysinecontaining peptides and, after 24 h, peptide depletion were analyzed by HPLC; Key-event (2): human HaCaT
keratinocytes were exposed to non-cytotoxic concentrations of HDIs for 24 h and intracellular IL-18 level was
evaluated (Corsini et al., 2013); Key-event (3): Dendritic cell-like U937 human cells were exposed to noncytotoxic concentrations of HDIs for 45 h and CD86 expression were evaluated (Natsch et al., 2013). RESULTS
AND DISCUSSION In DPRA assay, the HDIs were classified as sensitizers with an accuracy=80%, in
comparison with its classification based on human data, contrasting with the LLNA assay that reached an
accuracy of 60%. In relation to HaCaT keratinocytes-associated IL-18 and U937 assays, only one test-substance
was misclassified, and both assays showed accuracy=90%, when compared to human data. CONCLUSIONS
Based on key-events 1-3 of the SS, the strategy used here was able to identifying allergenic potential of HDIs
with accuracy superior to that obtained in the LLNA assay. Further studies are in progress to confirm these data
and to investigate the performance of this strategy, based on key-events of SS for hazard evaluation of HDIs
mixtures.
Financial Support
This work was supported by Fundação de Amparo à Pesquisa do Estado de Goiás (FAPEG), Fundação de Apoio à Pesquisa (FUNAPE) from Federal University
of Goiás; Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq); Financiadora de Estudos e Projetos (FINEP); and Coordenação de
Aperfeiçoamento de Pessoal de Nível Superior (CAPES).
380
PHYTOCHEMICAL CHARACTERIZATION OF Physalis angulata JUICE: PILOT STUDY
Gabriela Tassotti Gelatti
Tamiris Felippin
Ana Caroline Tissiani
Mariana Spanamberg Mayer
Nidia Ledur Muller Castro
Natacha Cossettin Mori
Diego Pascoal Golle
Jana Koefender
Introduction: The Physalis angulata is increasingly used in folk medicine as an anticoagulant,
antileukemic, antimutagenic, anti- inflammatory, antispasmodic, antiseptic and antidiabetic. However, little is
known about the antioxidant activity of this plant. Thus, the aim of this study was to investigate phytochemical
characterization and the antioxidant potential of Physalis angulata juice. Methods: This is a pilot study with the
fruit of Physalis angulata, which was grown in the University Campus of UNICRUZ in northwestern Rio Grande
do Sul. For the production of juice, fruits were harvested at the end of maturation and weighed (250 g), followed
by milling of the fresh fruit, dilution with water for the concentration of 250 g/L and lyophilization. Measurement of
total polyphenols was performed by the method described by Chandra and Mejia (2004) and condensed tannins
were measured by the method described by Morrison et al. (1995). Results and Discussion: There was a
concentration of 3.99 ± 0.24 mg/g of total polyphenols and 1.78 ± 1.68 mg/g of condensed tannins in Physalis
angulata juice at a concentration of 250 g/L. The total polyphenol concentration was lower than that found by
Vargas et al. (2008) in grape juice (440.11 mg/L). Similarly, the content of tannins was lower when compared to
the study by Rocha et al. (2011) in Campomanesia sp juice (81 ± 4 mg/g). Conclusions: The results demonstrated
that the juice from Physalis angulata has low concentrations of antioxidant compounds in the composition, thus it
becomes necessary to perform in vivo and in vitro studies to confirm its antioxidant potential.
Financial Support
FAPERGS
Ethical approval
15510413.3.0000.5322
381
OXIDATIVE STRESS CAUSED BY TAMOXIFEN DURING THE GLUCONEOGENESIS IN
PERFUSED RAT LIVER
Suélyn Koerich
UNIOESTE - University of Western Paraná
Suélyn Koerich
Fernanda Coleraus Silva
Carla Brugin Marek
Ana Maria Itinose
CEATOX - Assistence Center in Toxicology
Fabiana Sari Ferreira
Priscila Da Caz
INTRODUCTION: Tamoxifen is classified as a selective modulator of estrogen receptor (SERM) used
in the treatment and prevention of breast cancer. Although it has potential therapeutic effects, your side effects
are intense and are related to different mechanisms of toxic action, such as changes in the electrons flow of the
respiratory chain and changes in glycolytic pathway, causing oxidative stress, lipid peroxidation, and
consequently serious liver damage. Therefore, this study was designed to evaluate the tamoxifen influence on
hepatic oxidative stress during the gluconeogenesis. METHODS: Hepatic perfusion were performed in the male
wistar rats livers; we used fasted rats for the study of gluconeogenesis using glycerol 5µM of substrate of this
glycolytic pathway. Tamoxifen 100 µM with glycerol 5µM was perfused (TAM group); a group which was perfused
only glycerol was added as a control group (CON group). After perfusion, liver homogenate was prepared. The
dosage of thiols (Ellman et al., 1961), lipid peroxidation (Lpx) (Ohkawa et al., 1979) and glutathione (GSH) (Hissin
and Hilf, 1976) was measured (Significative p < 0.05). RESULTS AND DISCUSSION: The results show a
statistically significant decrease of GSH (p = 0.0298). Although, there was no significant variation in thiols (p =
0.0556), it was observed 26,55% of decrease in the TAM group compared to CON group, demonstrating the
antioxidant capacity expensed because GSH and thiols are part of system of cellular defense. A statistically
significant increase was showed in the Lpx (p = 0.0013) of TAM group over the CON group, confirming the
oxidative stress induction by tamoxifen during the gluconeogenesis. CONCLUSIONS: These findings confirm that
tamoxifen may cause oxidative stress and lipid peroxidation. Justified by multiple adverse effects of tamoxifen
and its ability to induce the formation of species reactive oxigen, leading to liver cell damage.
Financial Support
Higher Education Personnel Improvement Coordination (CAPES).
Acknowledgments
This work was supported by Laboratory of Cellular Toxicology and The Master's program in Pharmaceutical Sciences from University of Western Paraná.
Ethical approval
Ethics Committee in the use of animals (CEUA) from de State University of Maringá (Number: 036/13)
382
EVALUATION OF THE ESTROUS CYCLE OF RATS TREATED WITH CRUDE EXTRACT
OF Limonium brasiliense
Tânia Mara Antonelli Ushirobira
Andressa Blainski
Flávia Vitorino Constantino
Eneri Vieira de Souza Leite Mello
Introduction: Treatments with medicinal plants for female reproductive health has gained ground among
by women. Objective: This study aimed to evaluate, in an experimental model in vivo, the hormonal activity of the
crude extract (CE) from rhizomes of Limonium brasiliense, a brazilian biodiversity plant, used in southern Brazil
for treating menstrual disorders. Methods: Immature female Wistar rats, with 21 days old, were divided into
groups (n=6) which received by gavage for 28 days, CE at different doses (12.5, 25, 50, 100 or 200 mg/kg), water
(NC), and 400 µg/kg of βestradiol 17-valerate (PC). The following parameters were assessed: body weight, feed
intake, vaginal canal aperture, vaginal secretion cytology to determine the estrous cycle phase, uterotrophic
analysis, estrogen and progesterone dosages, and histology of the reproductive organs, uterus, ovaries, and
vagina. The experimental procedure was approved by the Ethics Committee on the Use of Experimental Animals
of UEM (Number 064/2011). Results and Discussion: The results showed no statistical difference among the
groups for body weight assessment and feed intake. The evaluation parameters of hormonal activity showed an
increased in days to the age of the vaginal canal aperture in all groups treated with CE compared to the PC group.
The analysis set: a) increase of the relative uterus weight; b) the estrous cycle predominantly in the diestrus
phase; c) the estrogen and progesterone dosages related to the estrous cycle phase; e) histological analysis of
the organs uterus, ovaries, and vagina, showed a tendency of progesterone action in a possible interference of
estrogen action at doses of 25, 50, or 100 mg/kg. Conclusion: The results point to a possible effect on the female
reproductive system, and it is suggested relationship with the popular use of Limonium brasiliense as a hormonal
regulator.
Financial Support
CAPES, CNPq, INCT_if, Pronex/Fundação Araucária.
Ethical approval
Number 064/2011
383
CIPROFLOXACIN ANTIBACTERIAL EFFICACY AGAINST Pseudomonas aeruginosa
BIOFILMS AND PLANKTONIC SHEDDING CELLS
Victória Etges Helfer
¹Faculty of Pharmacy, Federal University of Rio Grande do Sul, Porto Alegre, Brazil
Bruna Gaelzer Silva Torres
²Pharmaceutical Sciences Graduate Program, Federal University of Rio Grande do Sul, Porto Alegre, Brazil
Alexandre José Macedo
²Pharmaceutical Sciences Graduate Program, Federal University of Rio Grande do Sul, Porto Alegre, Brazil
Teresa Cristina Tavares Dalla Costa
¹Faculty of Pharmacy, Federal University of Rio Grande do Sul, Porto Alegre, Brazil; ²Pharmaceutical Sciences Graduate Program, Federal
University of Rio Grande do Sul, Porto Alegre, Brazil
Introduction: The chronic nature of some Pseudomonas aeruginosa infections is in part due to its ability
to form biofilms which confers intrinsic resistance to drugs, allowing the bacteria to survive to concentrations
hundreds of times higher than the concentrations able to kill planktonic cells of the same strain. We evaluated the
time-kill curves of ciprofloxacin (CIP) on P. aeruginosa biofilms aiming to compare the susceptibility of planktonic
and biofilm growing cells. Methods: Minimal Inhibitory Concentration (MIC) of CIP against P. aeruginosa ATCC®
28753 was determined by the broth microdilution method (CLSI M07 - A9). Minimal Biofilm Inhibitory
Concentration (MBIC) was determined following REITER et al. (Diagn Microbiol Infect Dis, 2012). For the time-kill
curves construction, P. aeruginosa biofilms were previously formed on the top of acrylic blocks. The blocks were
placed in culture flask containing 40 mL of Müeller-Hinton (MH) broth and exposed to several CIP concentrations,
multiples of MIC and MBIC, ranging from 0.0625 to 1000 µg/mL. Aliquots of the broth (planktonic cells) and a
block (biofilm cells) were sampled at different time points up to 24 h and bacteria were counted after processing
and incubation at 37 ± 1º C. Results and Discussion: The MIC and MBIC determined were 0.125 and 0.5 μg/mL,
respectively. Planktonic cells were eradicated with concentrations above 0.5 µg/mL, while, for biofilm, twice this
concentration was needed (above 1 µg/mL). Interestingly, with CIP concentrations higher than 10 µg/mL a
decreased efficacy was observed, which can be explained by the paradoxical effect reported for quinolones.
Conclusions: Higher CIP concentrations are needed to eradicate biofilm cells (2 x MBIC) while the planktonic
cells were eradicated with concentrations four times the MIC (1 x MBIC). The observed paradoxical effect
indicates that CIP has an optimum bactericidal concentration, highlighting the need to elucidate its efficacy in
vivo.
Financial Support
CNPq/Brazil (Grant 480366/2012-8)
Acknowledgments
Scholarship from CNPq/Brazil and CAPES/Brazil
384
CYTOTOXICITY AND APOPTOSIS INDUCTION OF IMIDAZACRIDINE DERIVATIVE
LPSF/AC05 IN BREAST CANCER CELLS
Valécia de Cassia Mendonça da Costa
Ivan da Rocha Pitta
Introduction: Breast cancer is the most frequent type of cancer in female population and presents a high
mortality rate. Many currently available treatments have a low therapeutic index, especially in triple-negative
breast cancer (TNBC) cases. Thus, this study aimed to evaluate the potential of LPSF/AC05, an imidazacridine
derivative, for breast cancer therapy. Methods: Breast cancer cell lines T47-D (ER/PR+), MCF-7 (ER+), MDAMB-231 (TNBC) and MCF-10A (non-tumoral breast cells) were used for tests. Cytotoxicity assays were
performed by MTT method, in which cells were exposed to AC05 (1 to 75DM) for 72 hours. Non-treated cells,
doxorubicin and DMSO (0,1%) were used as controls. Analysis of the potential induction of cell death was
performed using Apo-BrdU Apoptosis Detection kit. Cells were treated with AC05 at IC50 values during 48 hours
and analyzed by flow cytometry. Statistical analysis were performed using Student's t-test and differences were
considered significant when p<0.05. Results and discussion: AC05 showed cytotoxicity against MDA-MB231 and
MCF-7 (IC50 = 27.54 and 42.03DM, respectively). Although AC05 showed an IC50 value of 28.53 µM in MCF10A, it was still 30 times less toxic than doxorubicin (IC50<1µM). AC05 treatment significantly increased the
percent of apoptotic cells when compared to untreated cells in MDA-MB-231 (0.8 to 48.95%, p=0.0002) and MCF7 (1.1 to 28.52%, p=0.0018) and also compared to doxorubicin (p=0.001 and 0.0025 respectively). AC05 did not
significantly increased the percent of MCF-10A, whereas doxorubicin showed significantly increased of apoptotic
cells when compared to untreated cells (p=0.0113) and compared to AC05 treatment (p=0.016). Conclusions:
AC05 showed toxicity and apoptosis induction in two breast cancer cell lines, mainly in triple negative cells. It
showed lower toxicity in non-tumor breast cells when compared to doxorubicin, showing its selective potential.
Financial Support
FACEPE, INCT_if.
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