editorial board...v brazilian journal of pharmaceutical sciences the 53th pharmacy and biochemistry...

Editorial BoardThe Editorial Committee, which determines the editorial policy of the BJPS, is composed of four members three of them elected by Faculty of Pharmaceutical Sciences of the University of São Paulo.

Editorial Office

Brazilian Journal of Pharmaceutical SciencesDivisão de Biblioteca e Documentação do Conjunto das Químicas da USP • Serviço de Publicações e CirculaçãoP.O. Box 66083 • 05315-970 • São Paulo, SP • BrasilTel +55 11 3091-3804 • Fax +55 11 3867-8627E-mail: [email protected]

Proof and ReprintsThe author who submit the manuscript receive them from the Editorial Office. The proofs must be corrected and sent back up to 48 hours.

Subscription Brazil Outside BrazilInstitution R$ 350,00 US$ 200,00Personal R$ 175,00 US$ 100,00

GiftOnly to public organizations of teaching and research.

ExchangeWith those journals of interest to the Library of Conjunto das Químicas/USP.

Brazilian Journal of Pharmaceutical Sciences, São Paulo, v.53, suppl.1, (2018) – São Paulo: Faculdade de Ciências Farmacêuticas, USP, 1999- v., il.; 28cm.

Trimestral Continuação de: Revista Brasileira de Ciências Farmacêuticas/ Brazilian Journal of Pharmaceutical Sciences, São Paulo, v.35, n.1, (1999). ISSN1984-8250

1. Farmácia. I.Universidade de São Paulo. Faculdade de Ciências Farmacêuticas 615 CDD

Back IssuesPreviously published volumes can be available directly from the Editorial Office.

Copyright © 2008 - Faculdade de Ciências Farmacêuticas da USP. All rights reserved.

When submitting a manuscript, the authors agree with the transference of the copyright to the Publisher. The copyright holder’s consent comprehends the right of reproducing and distributing papers, photographic, microform or any other kind of similar reproduction, besides translation. It does not include distribution of copies for any purpose and by any means, unless a written permission from the Publisher is obtained.

Production DetailsPrinted on acid-free paper.

The Brazilian Journal of Pharmaceutical Sciences is edited quartely by the Faculty of Pharmaceutical Sci-ences, University of São Paulo and it continues the “Revista Brasileira de Ciências Farmacêuticas/Brazilian Journal of Pharmaceutical Sciences”, which has its origin in the “Anais de Farmácia e Odontologia da USP”, that began on 1939.The Brazilian Journal of Pharmaceutical Sciences publishes research papers and reviews in all fields of Pharmaceutical Sciences and it reflects the advance that has been made in this area.It is mandatory that manuscripts submitted to BJPS have neither yet been published nor submitted simultaneously to any other journal. It is worth to note that the contents of the papers are exclusively the authors responsibility.

CREDENCIAMENTO E APOIO FINANCEIRO DO PROGRAMA DE APOIO ÀS PUBLICAÇÕES CIENTÍFICAS PERIÓDICAS DA USP - 2008

Finnancial Support

On line versionhttp://www.bjps.br

http://www.scielo.br

XXIII PHARMACEUTICAL SCIENCE AND TECHNOLOGY MEETING OF THE FACULTY OF PHARMACEUTICAL

SCIENCES, UNIVERSITY OF SÃO PAULO10th ANNUAL RESEARCH SYMPOSIUM ON

PHARMACEUTICAL SCIENCES

ABSTRACTS

October 22-31, 2018

São Paulo – Brazil

v

Brazilian Journal ofPharmaceutical Sciences

The 53th Pharmacy and Biochemistry University Week (SUPFAB) is a pioneer annual event that was created in 1965 by the students of the academic center of pharmacy-biochemistry of the School of Pharmaceutical Science of University of São Paulo (FCF/USP). In 1995, the SUPFAB joined the Pharmaceutical Science and Technological Meeting (SFCT), an event organized by the research and the post-graduation committees of the FCF/USP. We have already established 23 years of a fruitful partnership between students and professors that has brought excellent results so far. Respecting each category sovereignty, everyone acquires more experience and is rewarded by updated knowledge gain during the whole process. This is worthy and we hope it remains for the next events. Over the years, we could conciliate various themes of general interest to students from different levels of knowledge. Lectures and courses were addressed to include classic and fundamental themes of the pharmaceutical profession to themes that are in the frontiers of knowledge. Students present their work in oral or poster sessions in which brings a unique opportunity for learning and sharing information developed in FCF/USP. The traditional XVIII diabetes, hypertension and hyperercholesteronemia campaign was created and coordinated by Professors Mario Hiroyuki Hirata and Rosario Dominguez Crespo Hirata. Patients from internal and external university community were assembled and received guidance and laboratory checkup. All students have the opportunity to meet and talk to the patients, improving their experience as future professionals and citizens. We would like to thank the dedication and commitment of all organizers, the academic center of pharmaceutical biochemistry, all students, employees and staff of all divisions of the FCF/USP, which make this week possible. We also thank all the speakers who have shared their knowledge and experience with us.

Prof. Dr. Joilson de Oliveira MartinsPresident of the Scientific Committee

Prof. Dr. Primavera BorelliDirector - Faculty of Pharmaceutical Sciences

PREFACE

vi


XXIII PHARMACEUTICAL SCIENCE AND TECHNOLOGY MEETING OF THE FACULTY OF PHARMACEUTICAL SCIENCES, UNIVERSITY OF SÃO PAULO

President of the Scientific Committee: Prof. Dr. Joilson de Oliveira MartinsVice-President of the Scientific Committee: Prof. Dr. João Paulo Fabi

Members: Profa. Dra. Cristina Helena dos Reis SerraProfa. Dra. Gisele Monteiro de SouzaProfa. Dra. Lygia da Veiga Pereira (IB/USP)Profa. Dra. Catarina da Fátima Pereira Teixeira (Instituto Butantan)Eduardo Alves (Assistente Técnico Acadêmico)Cleonice Estrela Cabral Gonçalves (Assistente Técnica Financeira)Yara Maria Lima Mardegan (Assistente Técnica Administrativa)

Committee for Evaluation of Abstracts and Oral Presentations Prof. Dr. Jorge Mancini Filho (FBA, Chairman)Prof. Dr. Uelinton Manoel Pinto (FBA)Profa. Dra. Sabrina Epiphanio (FBC)Prof. Dr. Eduardo Lani Volpe da Silveira (FBC)Prof. Dr. Gustavo Henrique Goulart Trossini (FBF)Prof. Dr. Humberto Gomes Ferraz (FBF)Profa. Dra. Susana Marta Isay Saad (FBT)Profa. Dra. Juliana Neves Rodrigues Ract (FBT)Prof. Dr. Marcelo Macedo Rogero – FSP-USPProf. Dr. Marco Aurélio Ramirez Vinolo – IB/UNICAMP

Committee for Evaluation of Poster Presentations Prof. Dr. Jeanine Giarolla Vargas (FBF, Chairman)Prof. Dr. Christian Hoffmann (FBA)Profa. Dra. Lígia Bicudo de Almeida Muradian (FBA)Profa. Dra. Ana Paula de Melo Loureiro (FBC)Profa. Dra. Carla Taddei de Castro Neves (FBC)Profa. Dra. Elizabeth de Souza Nascimento (FBC)Profa. Dra. Dominique Corinne Hermine Fischer (FBF)Prof. Dr. Gabriel Lima Barros de Araújo (FBF)Prof. Jarlei Fiamoncini (FBA)Profa. Dra. Cristina Northfleet de Albuquerque (FBT)Profa. Dra. Marina Ishii (FBT)Profa. Dra. Alice Cristina Rodrigues – ICB-USPProf. Dr. Fernando Rodrigues de Moraes Abdulkader – ICB-USP

vii


10th ANNUAL RESEARCH SYMPOSIUM ON PHARMACEUTICAL SCIENCES

Symposium of the Graduate Program in Food ScienceCoordinators: Prof. Dr. João Roberto Nascimento, Prof. Dr. João Paulo Fabi, Dra. Samira Bernardino Ramos do Prado

5th Symposium Drugs Planning and Development Symposium for Neglected DiseasesCoordinators: Profa. Dra. Jeanine Giarolla Vargas and Prof. Dr. Roberto Parise Filho

SECRETARY Maria Claudia PernaElza Silva GoesShirley Bornia MachadoKrisley Bornia Ghilardi

FINANCIAL SUPPORT Foundation for Research Support of São Paulo (FAPESP) Coordination of Higher Education (CAPES) – PROEXGraduate program in Pathophysiology and ToxicologyGraduate Program in Food Science


XXIII Pharmaceutical Science and Technology Meeting of the Faculty of Pharmaceutical Sciences,

University of São Paulo

ABSTRACTS

Brazilian Journal of Pharmaceutical Sciencesvol. 54, suppl. 1, 2018 3

FCF001-2018CUBOSOMES AS NANOPARTICLES FOR DRUG DELIVERY, BIOPHYSICAL CHARACTERIZATION AND INTERACTION WITH A MODEL DRUG: MILTEFOSINE

BÁRBARA MALHEIROS (M), RAPHAEL DIAS DE CASTRO (M), MAYRA CRISTINA GOMES LOTIERZO (M), GIOVANA FIRPO RODRIGUES (IC) LEANDRO RAMOS SOUZA BARBOSA

Department of Pharmaceutical and Biochemical Technology, FCF/USP

Introduction and Objectives: Biophysical study of a nanoparticle called cubosomes (CUB) in water, 2.25% glycerol media and PBS buffer, morphological characterization of the colloidal system and interaction with varied quantities of a drug, miltefosine (MILT). Material and Methods:Phytantriol (PHY), a lipid, self assembles in cubic structure when in excess of water. In order to make CUB nanoparticles a polymer, pluronic F127 (F127) is added to provide colloidal stability. CUB dispersion was prepared from a bottom up approach. In summary, 100 mg of PHY were solubilized in ethanol and drop-wised in a solution of F127 at 1.11 mg/ml. The mixture was kept under magnetic sitting for 15min at 45°C, then it was brought to a rotary evaporator in order to eliminate the ethanol. The final sample volume was 5ml. MILT was incorporated into cubosomes from 0.5%w/w to 20%w/w (~0.2mM to ~10mM, respectively). Samples were analyzed fresh and over time.Results and Conclusions: From SAXS experiment, the cubosomal dispersion presented Pn3m cubic structure. When loading small amounts of MILT (up to 5%w/w), there was no change of crystallographic structure but it was found that the drug enlarges the water channels of the CUB in 3 nm, when loading 10%, 15% and 20% w/w of MILT the nanoparticles assumed a new cubic structure: Im3m. TEM and Cryo-EM revealed particles in varied sizes with internal structure in coexistence with liposomes and micelles. DLS analysis revealed nanoparticles of approximate 350 nm and moderate polydispersion, MILT does not influence significantly particle size. Samples were also submitted to lyophilization and extrusion, no structural changes occurred with the nanoparticles. Financing: FAPESP; CNPq

FCF002-2018SYNTHESIS OPTIMIZATION OF TWO ACTIVE PHARMACEUTICAL INGREDIENTS APPLYING PROCESS INTENSIFICATION

GABRIELA CONSOLINI (M), CAROLINE ARASKIRO PESSOA (IC), MAURI SERGIO ALVES PALMA

Department of Biochemical and Pharmaceutical Technology, FCF/USP

Introduction and Objectives: Diabetes cases increased in Brazil in the last decade and have been encouraging the search for new drugs. Process intensification is important in the search for less harmful process and safer reactions; an example is the application of microreactors. In this work, the application of capillary microreactors is studied in the synthesis of (Z)-5-(4-hydroxy-3- methoxybenzylidene)2,4-thiazolidinedione (HMTZD) and (Z)-5-(3-hydroxy-4-methoxybenzylidene)2,4-thiazolidinedione (MHTZD), obtained from the reaction of 2,4- thiazolidinedione (TZD) with Vanillin and Isovanillin, which can be used in the synthesis of molecules with biological activity.Material and Methods: Two solutions were prepared, 0.0667 M of TZD with 0.0267 M of pyrrolidine in 30 mL ethanol and 0.0667 M of aldehyde in 30 mL ethanol. They were pumped separately to the microreactor. The temperatures studied were 78, 98, 120, 140 and 160°C. Samples were collected in the mean residence times of 2, 4, 8, 12, 16, 20, 30 and 50 min. The reaction yield was obtained by HPLC-UV analysis.Results and Conclusions: In the HMTZD reaction, maximum product yield, 100%, was obtained at 160°C in only 50 min of reaction. In the MHTZD reaction, the maximum product yield obtained was 85% in 50 min at 120°C and did not show an enhance with the increase of temperature, suggesting that the reaction achieved the equilibrium. Considering the results discussed above, the advantage of the microreactor is evident, which facilitates the work with higher temperatures, promoting an increase in the process productivity with safety, being this one of the main advantages of the microreactor technology.Financing: FAPESP; CAPES; PUB-PRGUSP.


FCF003-2018EFFECT OF GLUTAMINE AND ALANINE SUPPLEMENTATION ON PLASMA AND TISSUE GLUTATHIONE OF RATS SUBMITTED TO RESISTANCE TRAINING

AUDREY COQUEIRO (D), RAQUEL RAIZEL (D), ANDREA BONVINI (D), MARCELO ROGERO (PD)*, JULIO TIRAPEGUI (PD)

Department of Food and Experimental Nutrition, FCF/USP, *Department of Nutrition, FSP/USP

Introduction and Objectives: Exhaustive physical exercise promotes oxidative stress, which contributes to muscle damage and fatigue, impairing the exercise performance; thus, nutritional strategies are applied to attenuate these situations. Evidence indicates that glutamine supplementation may increase the concentrations of glutamate, an amino acid essential to the synthesis of the most important non-enzymatic antioxidant in the cell, glutathione. Therefore, this study aimed to investigate the effect of glutamine and alanine supplementation in the concentrations of plasma and tissue glutathione of rats submitted to resistance training (RT). Material and Methods: Forty male Wistar rats (n=8/group) were submitted to eight weeks of RT, which consisted of climbing a ladder with progressive loads increase (25 to 100% of body weight). In the last 21 days of RT, animals were supplemented with glutamine and alanine, as dipeptide (DIP) or in their free form (G+A and ALA groups), or water (SED and CON groups). Plasma, skeletal muscle (tibialis anterior), liver and heart glutathione were analyzed by colorimetric kit (Sigma-Aldrich). Results and Conclusions: RT and amino acids supplementation did not affect plasma, liver and heart glutathione concentrations, but all trained groups (CON, ALA, G+A and DIP) presented higher levels of muscle glutathione, by approximately 40%, compared with sedentary animals (p=0.003). In conclusion, glutamine and alanine supplementation, in their free or conjugated form, did not affect the content of plasma and tissue glutathione, while RT increased muscle glutathione concentrations of rats after eight weeks of training.Financing:FAPESP

FCF004-2018REPORTED FUNDING INCREASES CLINICAL GUIDELINES QUALITY: A SYSTEMATIC ASSESSMENT OF METHODOLOGICAL QUALITY AND TRANSPARENCY

CAROLINE DE GODOI REZENDE COSTA MOLINO (D)1, NATHALIA CELINI LEITE SANTOS (M)1, FRANCIELE CORDEIRO GABRIEL (M)1, SHEILA KALB WAINBERG (M)1, LUCIANA PEREIRA DE VASCONCELOS (M)2, RAFAEL AUGUSTO MANTOVANI SILVA (M)2, DANIELA OLIVEIRA DE MELO (COORIENTADORA)2, ELIANE RIBEIRO (ORIENTADORA)1

1Department of Pharmacy, FCF, USP, 2Department of Pharmaceutical Sciences, UNIFESP

Introduction and Objectives:Clinical practice guidelines (CPGs) affect the quality of care, since they translate the scientific knowledge into evidence-based recommendations. However, the development of high quality CPGs is expensive. To date, few studies reported the impact of reported funding in CPGs quality. Aims: To estimate the impact of reported funding in the quality of CPGs.Material and Methods: This is a systematic review of CPGs published between 2011-2016. Detailed search strategies, eligibility criteria, and data extraction are published elsewhere (CRD42016043364). We conducted a comprehensive search in MEDLINE, Embase, and the Cochrane Library. Additionally, an extensive search was conducted on 12 specific CPGs websites. CPGs quality were assessed by 3 appraisers using the AGREE II instrument (6 domains). Discrepancies at any stage were solved by consensus or by another appraiser. Results and Conclusions: CPGs that reported funding had greater number of authors, applied an instrument to classify the supporting evidence, were updated, and performed systematic review. Contrary, CPGs that did not report funding had fewer number of authors, not classified the evidence, were the first version, and did not mention the methods to search the literature. CPGs that reported funding rated higher mean scores in all AGREE domains (p<.0001) when compared to CPGs that did not report funding, even after adjustments for potential confounders. Therefore, CPGs that reported funding were more likely to have higher quality. Financing: CAPES; CNPq; FAPESP


FCF005-2018GUT MICROBIOTA FROM PRETERM INFANTS SUBMITTED TO COLOSTRUM THERAPY

LUANA DO NASCIMENTO MOREIRA (M)*, ANDREA PENHA SPÍNOLA FERNANDES**, RUBÉNS FEFERBAUM***, CARLA TADDEI*

*Department of Clinical and Toxicological Analyses, School of Pharmaceutical Sciences, University of São Paulo, São Paulo, SP, Brazil; **Hospital Maternidade Leonor Mendes de Barros, São Paulo, SP, Brazil; ***Instituto da Criança do Hospital das Clínicas, University of São Paulo, São Paulo, SP, Brazil

Introduction and Objectives: Exclusive breastfeeding has an important role in neonatal development, and is important for the initial establishment of intestinal microbiota. Colostrum therapy is well tolerated even to the smallest newborns, and can decrease in parenteral nutrition and hospitalization time. Thus, the aims of present study are to characterize the intestinal microbiota of preterm newborns submitted to colostrum therapy.Material and Methods: Preterm infants from Hospital Maternidade Leonor Mendes de Barros, submitted to raw colostrum (RC, n=12) and pasteurized colostrum (PC, n=13). Stool samples were collected at: first evacuation (S1), 7º day of life (S2) and 22º (S3). The gut microbiota was characterized by 16S rRNA sequencing and quantitative real time PCR analysis. Results and Conclusions: The group RC had more abundance of Firmicutes, while PC had more abundance of Proteobacteria. At family level, RC group had more abundance of Clostridiaceae compared with PC group in the last two samples times. Total bacteria levels tend to increase over time in both groups and no difference in the amount of total bacteria between the groups were observed. The PC group tends to present larger amounts of Bifidobacterium compared to the RC group, and all infants had the presence this important genus for the initial establishment of gut microbiota. The gut microbiota profile from preterm infants submitted to colostrum therapy, even with pasteurized colostrum, showed similarities with published studies of healthy newborns on exclusive breastfeeding. Financing: FAPESP, CNPq.

FCF006-2018TRANS FATTY ACIDS IN CROISSANTS: NUTRITION ADEQUACY, LABELING AND LEGISLATION

MARINA CARDINOT MAFIOLETTI (IC), LUCIANA TEDESCO YOSHIME (PD), JOSÉ AUGUSTO GASPAROTTO SATTLER (D), ILLANA LOUISE PEREIRA DE MELO (PD), JORGE MANCINI FILHO

Department of Food Science and Experimental Nutrition

Introduction and Objectives: The excessive ingestion of trans fatty acids (TFA) has been related to the development of cardiovascular disease (CDV). Recent data show that the Brazilian population diet exceeds the recommendations of TFA intake, due to the high consumption of industrialized food, among then the croissants, biscuits, breads and other bakery products. The World Health Organization (WHO) recommends the elimination of TFA from the food chain until 2023. The aim is to determine and evaluate the composition of trans fatty acids of croissants. Material and Methods: Eight samples of croissants were marketed in the city of Sao Paulo, and collected information about nutritional table: amount per serving (g), total fat (g), saturated fat (g) and trans fat (g). The total fat content was determined by acid hydrolysis and the lipid profile by gas chromatography according official method with some adaptations. The results were compared to the nutrition facts.Results and Conclusions: The total fat show expected variation, however, on five samples the contents of TFA were high, and on two samples the contents of saturated fatty acids (SFA) were high, both varying more than 20% of the declared. The results of the composition disagree with the nutrition information on the label and current legislation. Inconsistencies such as these entail the underestimation of the actual intake of TFA and may exceed the recommended limit for daily intake without consonance of the consumer. To reduce the intake of TFA and control to CVD risk factors, it is necessary attention to the label, reduce the flexibility variation of nutritional information, standardize the portion to 100 g, and replace technologies used in the use of oils and fats.Financing: CNPq


FCF007-2018POLYPHENOLS FROM CAGAITA FRUIT (Eugenia Dysenterica DC) ATTENUATE HEPATIC INFLAMMATION AND GLUCONEOGENESIS IN DIET-INDUCED OBESITY

CARLOS MARIO DONADO-PESTANA (PD), LARISSA RODRIGUES (PG), ÉRIKA VICÊNCIA MONTEIRO PESSOA (PG), MARIA INÉS GENOVESE

Departamento de Alimentos e Nutrição Experimental, Faculdade de Ciências Farmacêuticas – Universidade de São Paulo

Introduction and Objectives: Polyphenol-rich cagaita (Eugenia dysenterica. DC) extracts (PCE) have previously shown to prevent body weight and adiposity induced by high-fat/high-sucrose (HFS) diet in mice. In face of its effectiveness to prevent the development of diet-induced obesity and complications, we raised the question whether PCE could also be beneficial in an already developed obesity.Material and Methods: Male C57BL/6J obese mice (previously feed with a HFS diet for six weeks) were treated with PCE at two doses, 7 mg gallic acid equivalent (GAE)/kg body weight (PCE I group), and 14 mg GAE/kg body weight (PCE II group) or water (HFS group) by oral gavage for eight weeks. One healthy group (lean mice) was fed with balanced chow diet and gavaged with water. After 14 weeks, mice were euthanized and blood and tissues including liver and adipose tissue (inguinal, retroperitoneal, epididimal and brown) were collected for analysis. Results and Conclusions: PCE did not affect body weight and adiposity of obese mice. However, PCE did protect against dyslipidemia, fasting hyperglycemia, and glucose intolerance, and attenuated both hepatic inflammation and gluconeogenesis as observed by the expression of tumor necrosis factor-? and transcriptional factor NF- ?B. These results indicate that PCE improves glucose homeostasis of obese mice by attenuating both hepatic inflammation and gluconeogenesis. Financing: FAPESP; CNPq

FCF008-2018NUTRITIONAL COMPOSITION AND CARACTERIZATION OF FLAVONOIDS AND PHENOLIC ACIDS OF MAMA-CADELA (Brosimum gaudichaudii; MORACEAE)

GRAZIELI BENEDETTI PASCOAL (PD)1,2,3, JÉSSICA AITA GIACOMOLLI (IC)1, JULIANA FREITAS CHIARETO (IC)1, DANIELLE OLIVEIRA BORGES (M)1, FLORENÇA MARIA BORGES (M)2,3, ERIC DE CASTRO TOBARUELA (D)2,3, ISABEL LOURO MASSARETTO (PD)2,3, LAIS MORO (D)2,3, SILVIA LETICIA RIVERO MEZA (D)2,3, EDUARDO PURGATTO2,3

1Federal University of Uberlândia, Faculty of Medicine; 2University of São Paulo, School of Pharmaceutical Sciences; 3Food Research Center (FORC), CEPID-FAPESP

Introduction and Objectives: Mama-cadela (Brosimum gaudichaudii) is a native fruit from the Brazilian Cerrado biome, which has an orange-yellow coloring, a sweet flavor and may be consumed in natura. Overall, it can be considered as a good source of dietary fiber and bioactive componds as others Cerrado’s fruits, such as pequi, jenipapo and jatobá, for example. The objective is to analyze the nutritional composition and characterize the flavonoids and phenolic acids of mama-cadela. Material and Methods: Liquid chromatography (LC) coupled to mass spectrometry (MS) and AOAC methods. Results and Conclusions: The mama-cadela supplied: 69.69% moisture, 3.25% proteins; 2.03% fats; 0.45% ash; 18.02% “available” carbohydrates (by difference); 1.23% soluble dietary fiber (DF); 5.71% insoluble DF; 6.94% total DF; 1021.17mg/100g (mg%) soluble sugars (glucose, fructose and sucrose); 1546.11mg% organic acids (malic acid, citric acid and tartaric acid); 18.21mg% carotenoids and 74.04mg% phenolic compounds. The characterization of flavonoids and phenolic acids showed the following compounds: 3-O-caffeoylquinic acid, trans-5-O- caffeoylquinic acid, 4-O-caffeoylquinic acid, cis-5-O-caffeoylquinic acid, hesperidin, quercetin-4’-glucoside and quercetin-3-O-glucuronide. As conclusion, the mama-cadela provided good nutritional quality, highlighting the total DF and bioactive compounds (carotenoids and total phenolics). Financing: FAPEMIG


FCF009-2018PHENOLIC JABOTICABA EXTRACT PREVENTS EXCESSIVE GAIN OF BODY WEIGHT AND WHITE ADIPOSE TISSUES OF OBESE MICE

MÁRCIO HÉRCULES CALDAS MOURA (D), DENISE MATOS DE OLIVEIRA (IC), MARIA INÉS GENOVESE

Department of Food and Experimental Nutrition, FCF/USP

Introduction and Objectives: Phenolic compounds from jaboticaba, a Brazilian native fruit, has been associated to several benefits to health, such as improving the glucose and lipid metabolism, reducing inflammation and preventing the excessive gain of body weight and fat depots. This work aimed to evaluate whether phenolic compounds from Sabara jaboticaba (Plinia jaboticaba(Vell.) Berg) (PCSJ), in the form of a phenolic extract (PRJE), can halt the body weight gain of mice with pre-established obesity.Material and Methods: Seventy-two male 8-wk-old C57BL/6J mice were randomly distributed in two groups: negative (Ch, 20 animals) and positive (HF, 52 animals) control group. The Ch group was fed with a standard diet for mice and the HF group was fed with a high-fat-sucrose (HFS) diet for 14 weeks. After this step, 10 animals of each group were properly euthanized and the white adipose tissues (WAT) were collected. The HF group was redistributed in three groups: HF, J50 and J100, each one with 14 animals. In the last step with duration of 14 weeks, the Ch group was fed with the standard diet and water, the HF group was fed with the HFS diet and water, the J50 and J100 groups were fed with the HFS diet and PRJE, obtained by solid phase extraction with octadecylsilane, at the dose of 50 and 100 mg gallic acid equivalent/kg body weight respectively. Water and PRJE were administered by daily gavage. After 28 weeks the animals were properly euthanized, and the WAT were collected.Results and Conclusions: The animals that received the PRJE, regardless of the dose, showed gain of body weight and WAT approximately 50% lower than the HF group. These results suggest that the PCSJ have antiobesogenic properties and could be used as adjuvant in treatment against obesity, avoiding greater weight gain. Financing: CNPq

FCF010-2018DEVELOPMENT OF ECO-FRIENDLY CHROMATOGRAPHIC METHODS FOR THE SEPARATION OF SEVEN ANTIHISTAMINES: APPLICATION IN THE ANALYSIS OF AZELASTINE IN NASAL SOLUTION

LUCAS MACIEL DA COSTA (IC), ANIL KUMAR SINGH

Department of Pharmacy, FCF/USP

Introduction and Objectives: Antihistamines are widely used to alleviate the symptoms caused by allergic disorders. Most of these drugs have zwitteriónicas and/or amphoteric characteristics, which confer analytical challenges. The aim of this work was to develop highly efficient and greener chromatographic methods for separation and quantification of seven antihistamines: Azelastine, Desloratadine, Ebastine, Fexofenadine, Ketotifen, Loratadine and Olopatadine. Material and Methods: The separations were obtained with Shimadzu® HPLC system, using a RP C-18 LUNA (150x4.6mm, 5µm) column. The mobile phase consisted of acetonitrile and acidified water in following portions: desloratadine (15:85 v/v), ketotifen and olopatadine (25:75 v/v), azelastine (30:70 v/v), fexofenadine (32:68 v/v), loratadine (35:65 v/v) and ebastine (45:55 v/v). The prototype method was fully validity according to the guidelines of International Council for Harmonisation of Technical Requirements for Pharmaceuticals (ICH) and RDC No. 166 of 2017 and applied in the analysis of the azelastine, in nasal solution.Results and Conclusions: The system suitability parameters were adequate for proposed methods. Highly efficient separations were obtained for seven antihistamines, with following retention time (minutes): azelastine (5.2), ketotifen (3.7), desloratadine (5.5), ebastina (6.1), fexofenadine (3.7), loratadine (4.1) and olopatadina (6.5). The proposed method for azelastine showed linearity from 10-60 µg/mL (r² = 0.9962). The relative standard deviation in inter and intra-day analysis was < 0.72%. A near 100% recovery support accuracy. The DL and QL limits were 1.09 and 3.32 µg/ml, respectively. All methods abide by the green chemistry principles, with total organic waste < 2.5 mL / analysis. Financing: Undergraduate Pro-Rectory of USP & FAPESP


FCF011-2018MEASUREMENT UNCERTAINTY IN THE PERFORMANCE EVALUATION OF LIQUID CHROMATOGRAPHY ANALYTICAL PROCEDURES

LUCIANA SEPAROVIC (M), FELIPE REBELLO LOURENÇO


Introduction and Objectives: Liquid chromatography is one of the main techniques used in pharmaceutical quality control procedures. Moreover, analytical procedures should be validated according to the criteria of national and international guides. However, because of measurement uncertainty, even if a procedure meets the required validation criteria, its performance may not be adequate for its application in conformity assessment. In other words, the procedure may not fit-for-purpose. Thus, the aim of this study was to evaluate the performance of liquid chromatography analytical procedures based on their measurement uncertainty.Material and Methods: Articles regarding the validation of cited procedures were selected. The uncertainties were estimated based on the results of analytical procedures validation (accuracy and precision). Then, the ratio between overall uncertainty and tolerance range (U/T%) was calculated.Results and Conclusions: It was noted that in most cases (79%), precision was the critical parameter in the final uncertainty, being more related to pharmaceutical dosage forms that require more steps in sample preparation, therefore, more sources of uncertainty. In general, these pharmaceutical dosage forms, such as cream, gel and ointment, had higher measurement uncertainties, often above the recommended target uncertainty. The U/T% presented to be above the recommended value for 59% of the analytical procedures. Therefore, most of the analyzed procedures did not fit-for-purpose, showing the importance of considering the measurement uncertainty as part of analytical procedures validation, since accuracy and precision values affect directly the measurement uncertainty. High values of measurement uncertainty will compromise the analytical procedures performance in conformity assessment and their applicability in routine analysis.Financing: CAPES

FCF012-2018DESIGN, SYNTHESIS AND EVALUATION OF ARYLSULFONYL-HIDRAZONES AS ACETYLCHOLINESTERASE INHIBITORS FOR ALZHEIMER’S DISEASE

JULIA OLIVEIRA BARRIOS (IC), ROBERTO PARISE FILHO


Introduction and Objectives: Alzheimer’s disease (AD) is a neurodegenerative disorder that corresponds to 70% of the cases of dementia worldwide. Its physiopathology is related to the degeneration of cholinergic neurons and thus, acetylcholinesterase inhibitors are currently the main approach for AD treatment. Since there are so few treatment options that have limited efficacy and several adverse effects, the search for therapeutic alternatives has become essential. In view of this, aryl-sulfonylhydrazone analogues from the p- fluorophenylsulfonylhydrazone, which previously had inhibitory activity of acetylcholinesterase enzyme, were designed and synthesized. Material and Methods: The compounds were designed through molecular modifications such as ring opening and substituents variation. Docking studies were perfomed in GOLD Suite software to get information about target-molecule affinity. The synthesis involved the nucleophilic substitution to obtain the sulfonylhydrazide intermediate and nucleophilic addition to obtain the sulfonylhydrazones. Compounds were purified by chromatography column and characterized by 1H and 13 C NMR and melting point.Results and Conclusions: Dockin studies showed that the best score values were related to methoxy and hydroxy groups on the ring, but the position of the fluorine atom was irrelevant in this aspect. These results were taken into account to guide the synthesis and twelve compounds were obtained with yields between 14-88%. The two steps reaction to obtain sulfonyl-hydrazones have proven to be effective. The inhibitory activity of the compounds on AChE will be evaluated by the Ellman quantitative colorimetric assay. They are expected to demonstrate good efficacy profiles in order to provide important results for subsequent research and to assist the AD treatment..Financing:


FCF013-2018STUDY OF THE SACCHAROMYCES CEREVISIAE L-ASPARAGINASE 1 EXPRESSION IN PICHIA PASTORIS GLYCOSWITCH STRAIN - POTENTIAL USE IN THE TREATMENT OF ACUTE LYMPHOBLASTIC LEUKEMIA (LLA)

A.O. VALENCIA (D) AND G. MONTEIRO


Introduction and Objectives: Introduction and Objectives: Acute lymphoblastic leukemia was the first malignant disease to respond to chemotherapy. Most of these therapeutic treatments are based on the use of the enzyme L-asparaginase (ASNase) as the biopharmaceutical inhibitor of the abnormal cells proliferation. The asparagine, in turn, is an essential element for the growth of leukemic cells, but not for the normal cells. Thus, the deficiency of this amino acid in the blood serum can lead to tumor cell death by apoptosis. In this work, we propose to perform the expression of L-ASNase 1 in the glicoswitch strain of P. pastoris and to evaluate the humanized glycosylation induced in this yeast with the objective of improving its stability and decrease its immunogenicity, aiming also a new alternative for potential therapeutic use in the treatment of acute lymphoblastic leukemia (LLA). Material and Methods: Expression of the enzyme Sc_ASNase1, using the parental strain SuperMan5 (his-)glicoswitch of Pichia pastoris. Insertion of the ASP1 gene of Saccharomyces cerevisiae into vector pJAG-s1. Intergration of vector pJAG-s1_ASP1 by the method of transformation in the genome of the strain. Determination of the activity and purification of the enzyme L-ASNase 1 using the method of Frohwein. Evaluation of the post transcriptional modifications in the structure of this enzyme and citoxicity assay in leukemic cell lines. Results and Conclusions: As results of this work, we expect to obtain an alternative source in the expression of the enzyme L-asparaginase 1 in the strain Glicoswitch and that the effects of humanized glycosylation on this yeast can increase its stability and decrease the immunogenicityFinancing: CAPES

FCF014-2018MUTANT L-ASPARAGINASE OF Dickeya chrysanthemi (Erwinia chrysanthemi) WITH BETTER BIOCHEMICAL PARAMETERS

IRIS MUNHOZ COSTA (D), DÉBORA CUSTÓDIO MOURA, ADALBERTO PESSOA JUNIOR, GISELE MONTEIRO


Introduction and Objectives: Treatment of Acute lymphoblastic leukemia (ALL) is performed with L-asparaginase (L-ASNase), an enzyme obtained from bacteria Escherichia coli and Dickeya chrysanthemi (Erwinia chrysanthemi). However, both formulations are associated with a high rate of adverse effects that compromise the efficacy of the treatment. In addition, here in Brazil, the lack of commercialization occurs since 2013. In this way, the development of mutant isoforms from commercially available bacterial enzymes can contribute to the reduction of adverse effects and be an alternative for national marketing. Material and Methods: Using the L-ASNase from D. chrysanthemi has created a library of mutants from random mutagenesis, and a double mutant was selected. The mutant protein has been expressed in E. coli BL21(DE3) through IPTG induction and purified by ion exchange chromatography. For characterization, the mutant proteoform has been incubated with 10% human serum; in the presence of cofactors (Zn,2+-Mg2+-Ca2+-K+); and the optimum pH was determined. For this, the specific activity measurement was conducted using the Nessler’s reagent, colorimetric method for identification of ammonia.Results and Conclusions: The mutant proteoform showed specific activity 50% greater than wild type, with optimum pH at 8.0 and the presence of cofactors decrease its activity by up to 40%; however, the presence of human serum increases its stability over time. Increased activity suggests that proteoform can be a better than commercial enzymes. Financing: FAPESP 2016/25896-5, grant (Projeto Temático) 2013/08617-7.


FCF015-2018FINDING STRUCTURAL ORDER IN DISORDERED SYSTEMS: AMORPHOUS SOLID DISPERSIONS CHARACTERIZED BY PAIR DISTRIBUTION FUNCTION METHOD

VINICIUS DANILO NONATO BEZZON (PD), GABRIEL LIMA BARROS DE ARAÚJO


Introduction and Objectives: Amorphous Solid Dispersions (ASD) are systems used to enhance the bioavailability of low-solubility drugs since ASDs may increase their dissolution rate . The complexity of the sample structures that includes the structural disordering requires the use of Synchrotron X-ray powder diffraction (XRPD) and Pair Distribution Function (PDF) analysis to identify possible interaction between drug and matrices yielding to comprehension of the parameters that influence the drug’s physical stability. In this work, ASDs systems composed of flubendazole (FBZ) and polymers were prepared, and aiming to evaluate the FBZ at intra and intermolecular levels, in order to obtain information of the structural coherence of the systems studied. Moreover, the influence of each matrix avoiding the recrystallization of the drug was also evaluated. Material and Methods: ASD systems composed of FBZ dispersed in hydroxypropylmethylcellulose and its derivatives (HPMCP and HPMCAS) were prepared by the spray-dryer technique. The high-energy X-ray patterns were collected on the Beamline 6-ID-D at the Advanced Photon Source, Argonne National Laboratory, USA, using a beam of energy 100.315 keV to obtain high Q-values. Results and Conclusions: The sensibility of this XRPD analytical technique provides a great comprehension of local structure ordering compared to the conventional XRPD technique. The results showed that HPMCP and HPMC-AS can stabilize the systems as an amorphous solid dispersion for a long period compared to the HPMC-E3 matrix, which makes them promising candidates for ASD systems. Also, the PDF method is an efficient technique to be used in new formulation screening to select the best one, as it was presented by this work. Financing:

FCF016-2018ANTIBODY RESPONSE AFTER VACCINATION OF MICE WITH TWO NEW PLASMODIUM VIVAX CIRCUMSPOROZOITE PROTEINS FUSED TO MUMPS VIRUS NUCLEOCAPSID

RODOLFO FERREIRA MARQUES (D), ALBA MARINA GIMENEZ (PD)*, JANAÍNA TENÓRIO NOVAIS (IC), IRENE DA SILVA SOARES

Department of Clinical and Toxicological Analyses, School of Pharmaceutical Sciences, University of São Paulo, *Federal University of São Paulo

Introduction and Objectives: Plasmodium vivax remains a potential cause of morbidity and mortality for people who live where it is endemic. Based on our previous works, we developed two new recombinant proteins containing the P. vivax circumsporozoite protein (CSP) sequences fused to the Mumps virus nucleocapsid protein, as a strategy to generate antibody responses against the three variants of CSP (VK210, VK247 and P. vivax-like). Material and Methods: Mumps nucleocapsid-fused proteins were used as antigens in the presence of two different adjuvants. C57BL/6 mice were immunized subcutaneously with 10 µg of each antigen in the presence of adjuvants Poly (I:C) (InvivoGen) or Montanide® ISA 720 (Seppic). The IgG antibodies against the three P. vivax CS variants were determined by ELISA in sera obtained from mice two weeks after each immunizing dose. Results and Conclusions: Our results established that it is possible to elicit high (>104) antibody titers to all three different allelic forms of P. vivax CSP using these formulations. These recombinant proteins could be good candidates for clinical trials aiming at the development of a universal vaccine against P. vivax malaria.Financing: Fapesp 2012/13032-5; INCTV; CAPES


FCF017-2018IN VITRO EFFECTS OF TLR4 AND GAL-3 INHIBITION IN HUMAN COLORECTAL CANCER CELLS TREATED WITH PAPAYA PECTIN

RODRIGO GUIMARÃES LOPES (M), JOÃO PAULO FABI

Department of Food Science and Experimental Nutrition, FCF, University of São Paulo, SP, Brazil

Introduction and Objectives: Papaya pectins are soluble dietary fibers that may exhibit antitumor activity. This is due to the inhibition of a pro-metastatic protein (GAL-3) or by stimulating necroptosis through interaction with the transmembrane receptor TLR4. The present project aimed to elucidate which of these proteins are most prominently affected by the papaya pectin through chemical and biochemical (CRISPR-Cas9) inhibition. Material and Methods: Total cell wall (TCW) purification from fruit pulp was conducted using fully ripened papayas. TCW was extracted with water generating the pectin-rich Water Soluble Fraction (WSF). WSF sugar composition was determined through HPAEC-PAD after full hydrolysis with 2,0 M trifluoracetic acid. Oligo/polysaccharides molecular weight analysis was conducted on HPSEC-RID after filtration through 0,45 µm membrane. The metabolic activity of HT29 and HCT116 human colorectal cancer cells (CRCC) was measured through MTT assay after 24, 48 and 72h treatments with: WSF; TLR4 inhibitor Resatorvid (Res) + WSF; GAL-3 inhibitor N-Acetyl-Lactosamine (NAcLac) + WSF; and Res + NAcLac + WSF. Results and Conclusions: TLR4 and GAL-3 concomitant chemical inhibition impaired cell viability, indicating that pectins are able to hinder cancer cell development, in vitro. This might have happened in a way not directly dependent on the proteins studied. However, WSF + NAcLac treatment showed the most prominent effect on WSF capacity to reduce, in vitro, the viability of CRCC. This could indicate that GAL-3 blockage plays an important role in enhancing papaya pectins activity on colorectal cancer cells viability. Financing: FAPESP; CAPES

FCF018-2018CRITICAL CONTRIBUTION OF SINGLE PHOTON EMISSION COMPUTED TOMOGRAPHY (SPECT/CT) THE DIAGNOSIS OF IN EXPERIMENTAL MALARIA-ASSOCIATED ACUTE RESPIRATORY DISTRESS SYNDROME

THATYANE DE CASTRO QUIRINO (M) SABRINA EPIPHANIO

Faculty of Pharmaceutical Sciences, University of São Paulo

Introduction and Objectives: Infections by Plasmodium sp. could cause severe malaria with respiratory complications, resulting in the development of Acute Respiratory Distress Syndrome (ARDS), which can be identified using Single Photon Emission Computed Tomography (SPECT/CT). Material and Methods: The study was approved by the Ethical Committee (protocol 05/2017, ICB-USP). Non-infected and infected male mice (DBA/2 lineage) were used, being the Images obtained with SPECT/CT. Parasitemia, survival and respiratory parameters (BUXCO) were analyzed during the study.Results and Conclusions: The survival curve showed that ARDS-developing mice die between the 7th and 12th dpi, with a 30% increase in parasitemia compared with HP-developing mice (ARDS vs HP p <0.05). ARDS-developing mice presented decrease in respiratory frequency and increase in the respiratory pause compared to those non-infected (NI vs. ARDS p <0.01). On the 7th dpi, ARDS-developing mice showed a 30% decrease in hyperinflated and normally aerated lung tissues (NI vs. ARDS p <0.01) and 60% increase in poorly aerated and non- aerated lung tissues (NI vs ARDS p <0.001). Concerning regionalization of pulmonary perfusion in ARDS-developing mice, we observed 40% decrease in total pulmonary volume in the middle and base of lungs (NI vs. ARDS p <0.01) and 60% decrease in total pulmonary perfusion (NI vs. ARDS p <0.001) with 40% increase in total lung weight in mL (NI vs ARDS p <0.001). Thus, quantitative analysis with SPECT/CT technique is fundamental to increase the diagnostic sensitivity of lung lesions demonstrating hydrothorax and increased vascular permeability during the development of malaria-associated ARDS. Financing: CNPq; FAPESP.


FCF019-2018EVALUATION OF Bacillus atrophaeus ATCC 9372 SPORES GROWTH USING RESIDUAL MEDIA OF PHOTOSYNTHETIC MICROORGANISM CULTURE

MARIA EDUARDA GONÇALVES LOUSADA (M), ÉVELLIN DO ESPÍRITO SANTO (IC), GABRIEL FERRARI DIAS (IC), ELEANE DE ALMEIDA CEZARI GOMES (D), MARINA ISHII


Introduction and Objectives: Microalgae and cyanobacteria are photosynthetic microorganisms that require light, carbon dioxide, salts and organic nutrients for their growth, using a large volume of water, which is discarded after cultivation. This residual media is plenty of nutrients that could be reused as substrate for microorganisms’ cultivation. Bacillus atrophaeus is a microorganism that provides resistant spores applied as bioindicators for sterilization processes. The aim of this work is to evaluate the growth and development of spores cultivated in residual media in order to make an appropriate use of water that would be discarded as a pollutant in the environment. Material and Methods: For growth and development of B. atrophaeus ATCC 9372 spores, two residual media were used: F2 Guillard and Bold (kindly provided by Laboratório de Biotecnologia Microalgal, FBT-FCF-USP). Amount of 100 mL of reuse media (supplemented or not with yeast extract) previously sterilized at 121 oC for 30 min, was transferred to Erlenmeyers of 250 mL, inoculated with 1 mL of B. atrophaeus suspension in order to reach a final concentration of 103 total cells /mL, covered with gauze and incubated into orbital shaker at 150 rpm /37 oC up to 6 days.Results and Conclusions: After 6 days, residual media did not promote cell growth (population from 1.25*103 to 1.05*104 cells/mL). Addition of yeast extract, resulted in a population of 1.12*107 spores/mL and 3.50*107 spores/mL for F2 and Bold media, respectively, showing that microalgae residual media is viable to grow and develop B. atrophaeus spores. Financing: CAPES.

FCF020-2018SYNTHESIS OF LOBEGLITAZONE INTERMEDIATE AIMING THE TRANSPOSITION FROM BATCH TO CONTINUOUS FLOW PROCESS

ANDRÉ LUIZ KALIL MARCONDES (IC); RENAN RODRIGUES DE OLIVEIRA SILVA (D); MAURI SERGIO ALVES PALMA.


Introduction and Objectives: Lobeglitazone is a drug of the glitazone class used in the treatment of type 2 diabetes mellitus due to its effect of lowering blood glucose levels, acting on the PPAR? receptor, reducing insulin resistance in the cell. PPAR? is activated by the action of the drug and performs functions such as reduction to insulin resistance, modification of adipocyte differentiation, and reduction of leptin levels, leading to increased appetite, reduction of interleukin level and increase in adiponectin protein level. Microreactor Technology offers a number of advantages for chemical synthesis when compared to batch processes, such as high surface-to-volume ratio, more efficient homogenization of the reaction mixture, leading to increased reaction rates, reactant conversion, yield and selectivity. This work aims to perform the synthesis of the second step of Lobeglitazone synthesis for subsequent transposition to continuous flow synthesis in a capillary microreactor. Material and Methods: 3 mmol of 4-chloro-6-(4-methoxyphenoxy) pyrimidine, 3 mmol of 2-(methylamino) ethanol in 30 mL of ethanol was added to a round bottom flask. This reaction was carried out at room temperature. After 24 h the reaction was done and chromatographic purification was carried out with mobile phase solution 1:1 hexane/ethyl acetate. Results and Conclusions: Yield of 98% by mass was obtained and its identity was confirmed by HPLC-MS analysis, where the mass of (m/z) 276.08 (M + 1) was obtained. In the next step, samples by time will be taken to determine the reaction time in the batch process and transpose the reaction to continuous flow synthesis in the microreactor, aiming to reduce the reaction time. Financing: FAPESP


FCF021-2018EXPRESSION OF EXTRACELLULAR RECOMBINANT L-ASPARAGINASE II FROM Saccharomyces cerevisiae WITH HUMANIZED GLYCOSYLATION IN Pichia Pastoris

HENRIQUE BIASOTO (M), GISELE MONTEIRO


Introduction and Objectives: L-asparaginase is an efficient tumor growth inhibitor, used in chemotherapy sessions against acute lymphoblastic leukemia with 80% total recovery of treated patients. The severe immunogenic effects of Escherichia coli’s enzyme led us to seek other sources of L-asparaginase production, among them S. cerevisiae and P. pastoris, the latter with great potential for extracellular expression, which would decrease the production cost. The proteins expressed by these microorganisms present hyperglycosylation, especially S. cerevisiae, resulting in an immunogenic effect. To overcome these drawbacks, we propose cloning and expressing S. cerevisiae’s L-asparaginase II fused with heterologous extracellular expression signaling in P. pastoris GlycoSwitch, which presents humanized glycosylation.Material and Methods: We used the circular polymerase extension cloning technique to build two different vectors, with two different signaling sequence, both fused with L-asparaginase II. Linearized vector was transformed in P. pastoris GlycoSwitch and the enzyme was expressed by inducing the AOX1 promoter with methanol. The enzyme was analyzed with SDS-PAGE electrophoresis and the asparaginase activity measured by hydroxylamine. The purification protocol was estabilished.Results and Conclusions: The P. pastoris strain was successfully transformed and L-asparaginase II fused with both signaling sequences was incorporated into genome. The extracellular medium presented asparaginase activity and the SDS-PAGE revealed a protein with approximately 45kDa, similar to native L-asparaginase II, however, the protein expressed is glycosylated. Financing: CAPES; FAPESP

FCF022-2018VANCOMYCIN SERUM MONITORING IN SEPTIC PEDIATRIC BURN FOR TARGET ATTAINMENT

VANESSA KASUBECK DE SOUZA(M)*, DAVID DE SOUZA GOMEZ(D)**, ELSON MENDES DA SILVA JR(D)**, JOAO MANOEL DA SILVA JR(D)**, NILO JOSÉ DUARTE(D)***, SILVIA REGINA CAVANI JORGE SANTOS(PD)*

*Clinical Pharmacokinetics Center, Pharmacy Department FCF USP **Burn Center, Division of Plastic Surgery and Burning ***Division of Central Laboratory HC FMUSP (Sao Paulo, SP - Brazil)

Introduction and Objectives: Vancomycin is recommended to treat bloodstream infections caused by gram-positive strains in septic ICU pediatrics. The AUC/MIC ratio (AUCss

0- 24/MIC) is the recommended index to evaluate vancomycin effectiveness instead of trough levels widely used in the past for dose adjustment. The present study aims to investigate if the empirical daily dose can reach the therapeutic target against gram-positive strains MIC 1mg/L for dose adjustment purposes in a pediatric burn patient in septic shock with renal function preserved versus renal dysfunction. Material and Methods: The protocol was approved by the ethical committee N.5484. HLF (M), 12yrs, 60kg, 155cm, total burn surface area (15%) and inhalation injury were investigated. The empiric daily dose recommended was prescribed 0.75g q12h versus 0.5g q24h, 1h pump infusion. Target was based on drug effectiveness index (AUCss

0- 24/MIC>400). The drug serum measurements were done by immunoassay.Results and Conclusions: Important changes on pharmacokinetics were found in both sets during the clinical course of septic shock. Dose regimen 0.75g q12h versus 0.5g q24h resulted in target attainment against MIC 2mg/L strains. Highlights in renal failure set must be considered, once the reduction of daily dose contributed to drug effectiveness-safety. Thus, vancomycin serum monitoring based on target attainment done in real-time permits an earlier clinical intervention and desired clinical outcome with cure of infection during antimicrobial therapy and safety. Financing:


FCF023-2018HPLC-RP TO DETERMINE THE ENCAPSULATION EFFICIENCY OF AN ANTI-CANCER DRUG IN NANOPARTICLES

FERNANDO KANEKO PRADO (M)*, ELIANA MARTINS LIMA**, MARIA SEGUNDA AURORA PRADO*

*Department of Pharmacy FCF/USP **Pharmaceutical Technology Laboratory, Faculty of Pharmacy/UFG

Introduction and Objectives: According to the World Health Organization (WHO), in 2012, approximately 14 million new cases and 8.2 million deaths from various types of cancer were reported. Imatinib widely used in leukemias and gastrointestinal neoplasms. The present work aims to develop and validate a method to determine the encapsulation efficiency of the imatinib drug in previously elaborated and characterized nanoparticles by HPLC. Material and Methods: 200 µL of nanosuspension were filtered on a 0.45 µm Millex filter, to 100 µL of the filtrate,1000 µL of ethanol PA was added to break the nanoparticles, then 900 µL of water were added and centrifuged for 5 min. The supernatant was quantified by HPLC. To calculate Encapsulation Efficiency was used Equation. A ZORBAX C18 (100x4.6 mm x 3.5 µm) column was used. The flow rate was 1 mL.min-1 and absorbance was monitored at 267 nm. The mobile phase was 60% ammonium acetate buffer and 40% acetonitrile. Results and Conclusions: Validation of the proposed method followed ICH. The proposed method showed good linearity, R2=0.9998, over a concentration range of 3.0 - 60.0 µm.mL-1 of imatinib. Detection and quantification limits were 0.80 µg.mL-1 and 2.92 µg.mL-1, respectively. Intra-day precision was less than 2%. The precision was expressed as percentage of relative standard deviation (RSD). The accuracy expressed as recovery was 98.85 ± 0.04 %. The encapsulation efficiency of imatinib in nanoparticles was 90%. The proposed HPLC method showed to be simple, linear and precise and also suitable for determination of imatinib in nanoparticles and in pharmaceuticals. Financing: CAPES

FCF024-2018DEVELOPMENT AND CHARACTERIZATION OF SOLID DISPERSIONS OF NICLOSAMIDE

MARIANA RIBEIRO GUBITOSO (IC)*, NÁDIA ARACI BOU-CHACRA*, FLAVIO MACHADO DE SOUZA CARVALHO**, GABRIEL LIMA BARROS DE ARAUJO*

* Departamento de Farmácia, Faculdade de Ciências Farmacêuticas, Universidade de São Paulo ** Departamento de Mineralogia e Geotectônia, Instituto de Geociências, Universidade de São Paulo

Introduction and Objectives: Niclosamide (NCL) is an effective anthelmintic agent that in recent years has been repositioned for use in the treatment of tumors and viral infections. Its low solubility in water, however, may compromise its effective use and performance in preclinical and clinical trials. The present project explores possible alternatives through the research and development of stable amorphous solid dispersions aiming the reformulation of this drug, to indicate ways to improve its solubility. Material and Methods: An exploratory screening NCL-polymer dispersions was performed by rotary evaporation, Differential Exploratory Calorimetry (DSC) and X-ray powder diffraction. Results and Conclusions: Melting enthalpy and crystallinity index for each dispersion were successfully used to determinate the miscibility and stabilization behavior of Povacoat, hydroxypropylmethylcellulose derivatives (HPMC, HPMC-AS, HPMC-P), hydroxypropylcellulose, soluplus and poloxamers, indicating ways to develop niclosamide formulations with enhaced dissolution and stability performance. Financing: n.a


FCF025-2018POLYMORPH SCREENING AND SOLUBILITY CHARACTERISATION OF LERCANIDIPINE HCL

ILIA ALEKSEEVICH REPIN (M)*, HUMBERTO GOMES FERRAZ*, SELMA GUTIERREZ ANTONIO**, GABRIEL LIMA BARROS DE ARAUJO*

*Departamento de Farmácia, Faculdade de Ciências Farmacêuticas, Universidade de São Paulo **Universidade Estadual Paulista Júlio de Mesquita Filho, Instituto de Química de Araraquara, Departamento de Físico-Química

Introduction and Objectives: Lercanidipine (LRC) is a highly lipophilic dihydropyridine calcium antagonist with very limited information about its polymorphism and the potential impact on solubility and stability. The present work was focused on performing polymorphism screening by applying solvent based methods, medium throughput approaches, and solid-state studies. Material and Methods: Form I and form II of LRC were successfully obtained and characterized by thermal analysis and X-ray powder diffraction and solubilities studies.Results and Conclusions: Melting point temperatures and values of melting enthalpies indicate a monotropic relationship between two forms, with form II being the stable one. Temperature-dependent solubility studies in organic solvents using Crystal16TM revealed significant differences between form I and II. Solution-mediated polymorphic transformation study was also performed to confirm the stability relationship between the two forms. Financing: Financial support from FAPESP (2015/05685-7) and CNPq (fellowship to I. A. Repin).

FCF026-2018SAFETY AWARENESS OF HERBAL PRODUCTS IN A HEALTH CARE CENTER

ALEXANDRA ALMEIDA HÜBNER (PG), FELIPE REBELLO LOURENÇO, POLIANA MORAES1 (IC), DANIELA BRAGA (IC), PAULO CHANEL FREITAS, EDNA MYIAKE KATO


Introduction and Objectives: The Unified Health System (SUS) started a policy on Integrative and Complementary Practices, encouraging interdisciplinary actions for health care that increased the demand for herbal products (HPs) in 161%, between 2013 - 2015. Therefore, this research aimed to analyze the information on therapeutic uses of HPs for human health, as well as their side effects (SE) among patients of a public health care center. Material and Methods: The survey was applied during a presentation about HPs for 66 patients of the Samuel Pessoa Health Center (FM-USP). The results were categorized in citation, therapeutic use, plant part, use value (UV), sex and age. The study was approved by the Human Research Ethics Committee - USP. Results and Conclusions: In total there were 245 citations for 76 medicinal plants employed for 60 health problems. Average subject ages were 57 ± 18 for women and 60 ± 9 for men, with 89% more women. The most cited species were: Cymbopogon citratus (UV = 0.51), Plectranthus barbatus (UV = 0.39), Mentha sp (UV = 0.28), Matricaria chamomilla (UV = 0.27), Melissa officinalis (UV = 0.16) and Pimpinella anisum (UV = 0.16). The HPs were used to treat digestion problems (21%), anxiety (18%), influenza (8%), insomnia (6%) and inflammation (4%). The most cited plant parts were leaves (56%), flowers (11%) and fruits (9%). Medical prescription accounted for 6.5% of the uses, indicating a high self-medication rate for HPs, with only 4.5% of the subjects mentioning some SE. Although SUS provides HPs to patients, fresh medicinal plants are preferred. Thus, the consciousness of the risks and benefits of fresh herbal medicinal preparations must be increased. Financing: 1Programa Unificado de Bolsas (USP)


FCF027-2018RAPID MICROBIOLOGICAL METHODS (RMMS) FOR EVALUATING THE ACTIVITY OF CEPHALOSPORIN ANTIBIOTICS EMPLOYING TRIPHENYLTETRAZOLIUM CHLORIDE

ALESSANDRO MORAIS SAVIANO (D), FELIPE REBELLO LOURENÇO


Introduction and Objectives: The microbiological agar diffusion method has been used to evaluate the potency of antibiotics since the discovery of penicillin in the 1930s, but it has as a limitation regarding the incubation time, generally from 18 to 24 hours. The objective of the present study was to develop, optimize and validate rapid microbiological methods (RMMs) for cephalosporin antibiotics using triphenyltetrazolium chloride (TTC) to reduce the incubation time of the assays.Material and Methods: Cephalosporin antibiotics (cefazolin, cefuroxime, ceftriaxone, and cefepime) and a cell viability indicator (TTC) were used in a factorial design, in which the inoculum suspension, incubation time, and percentage of the TTC solution were varied. The validation of RMMs employed MHA medium in a bilayer system, with a superior layer (5 mL) inoculated with 2% of Staphylococcus aureus (ATCC 6538) suspension, incubation time of 5 hours and 30 minutes at 37 ± 1 °C and the addition of 0.3% TTC solution in 1% agar, cefazolin and cefuroxime in concentrations from 15 to 60 µg/mL, ceftriaxone and cefepime in concentrations from 20 to 80 µg/mL. Results and Conclusions: The RMMs were selective, linear (for standard and sample curves, respectively), accurate, precise, robust and rugged. After validation, the RMMs were compared with conventional microbiological methods, and the results found by both methods were statistically similar, however, RMMs had advantages of reducing the incubation time from 22 hours to 5 hours and 30 minutes. Therefore, RMMs may be an alternative in the quality control of dosage forms containing cephalosporin antibiotics.Financing: FAPESP

FCF028-2018APPLICATION OF ANALYTICAL TOOL FOR GALENICAL PROCESS – THE USAGE OF MIXER TORQUE RHEOMETER TO EVALUATE THE INFLUENCE OF DRUG SOLUBILITY ON WET GRANULATION PARAMETERS

BRUNA RODRIGUES BELEM (IC), ERIKA KISHIMA (IC), JESSICA LINS DOS SANTOS (IC), MICHELE GEORGES ISSA (PD), HUMBERTO GOMES FERRAZ


Introduction and Objectives: Wet granulation consists in particles adhesion with a binder resulting in small agglomerates, which have improved physical characteristics, providing the tablet production. Mixer Torque Rheometer (MTR) evaluates the torque of the wet mass, which has a directly proportional relationship with the agglutination rate of particle and the consistency of the mass. The objective of this study was to evaluate the influence of drug solubility in wet granulation process through the rheological study using MTR. Material and Methods: The experiments were performed on a Caleva MTR rheometer. A 32 fractional factorial experimental was designed to assess different binders (Kollidon VA64, gellan gum CG-HA, alginate), their percentage in the formulation and the drug concentration. Metoprolol succinate (MS) and sodium diclofenac (SD) were used as model for high and low solubility drugs, respectively.Results and Conclusions: Torque for SD was higher than for MS, demonstrating the influence of the drug solubility in wet granulation. MTR provides the ideal conditions for wet granulation, but depending on the material the process is not feasible, as observed with 80% MS formulation. Granules with 50% of MS presented more friable than granules with 50% and 80% of SD. Microscopic analysis demonstrated a different aspect between SD and MS, which may be related to solubilization and subsequent recrystallization of MS. This study showed that MTR is an interesting tool, enabling the evaluation of the influence of drug solubility in wet granulation. Financing: DEINFAR/FCF/USP


FCF029-2018SYNTHESIS OF ROSIGLITAZONE: STUDY OF THE FIRST STEP IN BATCH PROCESS

PAULO VICTOR CUESTA CALVO (IC), RENAN RODRIGUES DE OLIVEIRA SILVA (PG), MAURI SERGIO ALVES PALMA


Introduction and Objectives: Rosiglitazone (Avandia®) is an important drug used to combat type 2 diabetes mellitus and its synthesis consists of four essential and subsequent steps. Batch reactors have several limitations in terms of mixing efficiency and the use of high temperatures and pressures. Microreactors presents several advantages over batch processes, mainly in terms of safety operating under extreme conditions, and increase of selectivity and reaction yield, due to the small distances in the micro channels. In the present work, it was studied the synthesis of 2-(methyl(pyridin-2-yl)amino)ethan-1-ol, an intermediate of Rosiglitazone, aiming the transposition to continuous flow capillary microreactor.Material and Methods: 2-chloropyridine (10 mmol; 1.00 eq.) and 2-(methylamine)ethanol (10 mmol; 1.00 eq.) were put into a Schlenck tube. The solution was heated to 120°C and left to react for 24 h. At the end of the reaction, the mixture was allowed to cool down to room temperature. The cooled solution was then poured into 25 mL water at 4°C with ammonium chloride (5.61 mmol). The aqueous phase was extracted three times with ethyl acetate (20 mL), subsequently washed with brine and dried over sodium sulfate. The product was obtained using rotary evaporation under reduced pressure. Results and Conclusions: The yield was 77.7% in mass and its characterization was performed using HPLC-MS analysis, proving the identity of the pure product (m/z) 153.02 (M+1). As next steps, samples in function of time will be taken to determine the reaction time and it will be made the reaction transposition for continuous flow using capillary microreactors. Financing: FAPESP

FCF030-2018DETERMINATION OF CO-ENCAPSULATION EFFICIENCY OF TUBERCULOSTATIC DRUGS IN PLGA NANOSPHERES BY CAPILLARY ELECTROPHORESIS

LOURDES MARCELA YATACO-LAZARO (M)*, FERNANDO K. PRADO (M)*, JESÚS SANTAMARÍA**, JAVIER A. FERNÁNDEZ**, MANUEL ARRUEBO**, VICTOR SEBASTIAN**, MARINA F. M. TAVARES***, MARIA S. A. PRADO*

*FCF/USP **University of Zaragoza, Spain ***IQ/USP

Introduction and Objectives: Despite being one of the oldest and most well-known infectious diseases, TB remains the second leading cause of death after AIDS. The aim of the present study was to develop and validate a fast, precise and inexpensive capillary electrophoresis (CE) method for simultaneous determination of rifampicin and isoniazid in PLGA nanospheres. these are the drugs of first choice in the treatment of TB Material and Methods: For the development of the nanospheres, emulsion/solvent evaporation technique was used. Separation was achieved using an uncoated fused-silica capillary of 50 µm i.d. with a total length of 30 cm, an electrolyte containing ethyl acetate, butan-1-ol, sodium dodecyl sulfate, isopropanol, 10 mmol.L-1 sodium tetraborate aqueous buffer at pH 9.2. Samples were injected at 0.5 psi for 5 s, the applied voltage was 30 kV and the detection was made by UV absorption at 207 nmResults and Conclusions: The proposed method showed good linearity (R2>0.99) over the concentration ranges from 192.00 µg.mL-1 to 288.00 µg.mL-1 for rifampicin and 160.00 µg.mL-1 to 240.00 µg.mL-1 for isoniazid. The precisions were lower than RSD 1.5%. The detection limits for rifampicin and isoniazid were 6.60 µg.mL-1 and 7.70 µg.mL-1, respectively and the quantitation limits were 20.01 µg.mL-1 for rifampicin and 23.34 µg.mL-1 for isoniazid. The encapsulation efficiency of nanospheres was 2,33 % and 14,75 % for rifampicin and isoniazid, respectively. The CE method can be applied to determine tuberculostatic drugs in pharmaceuticals and in nanoparticulate systems Financing: CAPES


FCF031-2018EVALUATION OF THE ANTIMICROBIAL ACTIVITY OF PEPTIDE PRODUCED BY BIFIDOBACTERIUM ANIMALIS

RONALDO DOS SANTOS LIRA(M), TAÍS MAYUMI KUNIYOSHI(PD), SABRINA DA SILVA SABO(PD), RICARDO PINHEIRO DE SOUZA OLIVEIRA


Introduction and Objectives: Some lactic acid bacteria have the ability to produce antimicrobial peptides called bacteriocins which could present wide application in food industry. The objective of this work is to characterize the antimicrobial activity of cell free supernatant(CFS) produced by Bifidobacterium animalis(BF)Material and Methods: BF was cultured in MRS medium supplemented with 1% L-cysteine and 0.3% sodium thioglycolate at 37 °C for 24 hours. In order to obtain the CFS, The culture was centrifuged for 25 min at 2795 x g, and the supernatant had its pH adjusted to 6.5 and heated at 80 °C for 10 min. The antimicrobial spectrum of the resultant CFS was tested against Listeria innocua 2711(LI), Carnobacterium maltaromaticum CECT 4020, Staphylococcus aureus CECT 239, Escherichia coli ATCC 25922. The proteinaceus nature of the antimicrobial compound in the CFS was verified using 5 mg/mL of tripsina, pepsin or papain. The partial purification of CFS was carried out through ammonium sulfate (AS)(10% to 60% ) preciptation. Each sample was evaluated in Tris-SDS-PAGE and the antimicobial activity via spot-on-lawn assay. Results and Conclusions: The CFS produded by BF showed antimicrobial activity against different Gram positive bacterium such as L. innocua 2711, C. maltaromaticum CECT 4020, S. aureus CECT 239. The protease test revealed the proteinaceus nature of the antimicrobial compound produced by BF. Furthermore, partial purification of the CFS demonstrated that 20% of AS was obtained a predominant band around 2-5 KDa with 3200 AU/mL of activity against LI. Higher antimicrobial activity was observed (up 102400 AU/mL) in the sample precipted with 60% of AS but Tris-SDS- PAGE revealed that the resultaint produtct of purification still contains many other proteins Financing: CAPES

FCF032-2018CLONING AND EXPRESSION OF L-ASPARAGINASE FROM Escherichia coli IN A Pichia pastoris STRAIN WITH HUMANIZED GLYCOSYLATION

GUILHERME MEIRA LIMA1 (IC), BRIAN EFFER1,2 (D), GISELE MONTEIRO1

1Department of Biochemical and Pharmaceutical Technology, School of Pharmaceutical Sciences, University of São Paulo, São Paulo– Brazil. 2Department of Chemical Engineering, Faculty of Engineering and Sciences, University of La Frontera, Temuco – Chile.

Introduction and Objectives: Currently, acute lymphoblastic leukemia (ALL) is the most frequent type of cancer seen in children. One of the available treatments for ALL is L-asparaginase (L-ASNase), enzyme that catalyzes the conversion of amino acid L-asparagine into aspartate and ammonia. However, this biopharmaceutical product has disadvantages, since it may induce immunogenic reactions and can be rapidly cleared from our organism by different elimination processes. Our objective is to produce a novel humanized glycosylated L-ASNase since this could decrease its immunogenicity.Material and Methods: Linearized expression vector was designed and has integrated into the yeast genome. Expression of the desired protein was induced by methanol. Protein expression was verified by SDS PAGE, and glycosylation by treatment with PNGase enzyme and mass spectrometry. Enzymatic activity was evaluated using hydroxylamine method and purification steps were performed using ion-exchange chromatography.Results and Conclusions: Our engineered Pichia pastoris strain was capable of expressing glycosylated L-ASNase as well as secreting it outer of the eukaryotic cell. The newly developed recombinant protein has shown enzymatic activity (specificity activity = 2,98U/mg) as confirmed by colorimetric assay. Purification method was successfully optimized. Mass spectrometry analysis confirmed glycosylation occupancy sites and presence of Man2-9GlcNAc2 glycans attached to the recombinant L-ASNase structure.Financing: FAPESP


FCF033-2018Streptococcus agalactiae: SEROTYPE DISTRIBUTION AND CORRELATION WITH VIRULENCE FACTORS AND ANTIMICROBIAL RESISTANCE

CILICIA NASCIMENTO (PG), RENAN MARTINS (IC), CARLA R. TADDEI

Department of Clinical and Toxicological Analysis FCF/USP

Introduction and Objectives: Streptococcus agalactiae or Group B Streptococcus (GBS) is considered the leading cause of early-onset newborn sepsis, its presence in microbiota from women is a concern during pregnancy and labor. Brazil uses international data to establish public strategies of prevention and treatment. The objective of this study is to access the Brazilian profile of GBS.Material and Methods: A total of 292 GBS strains were isolated from both pregnant and non-pregnant women. GBS was isolated from vaginal and anal swabs, urine and vaginal discharge. The strains were obtained from three laboratories’ strain banks. We performed the strains serotype determination by a Multiplex PCR reaction. The samples were tested through PCR for the following virulence genes: hyaluronidase, cytolysin/hemolysin and pilus 1, 2A and 2B, using correspondent primers. Antimicrobial resistance was assessed through the disc- diffusion method, for Penicillin, Clindamycin, Erythromycin, Cefepime and Vacomycin. Results and Conclusions: Serotypes Ia and V were isolated more frequently, a result seen in literature. Our study found a higher prevalence of serotype V in non-pregnant women, which is related to invasive diseases in non-pregnant adults. Serotype III is more abundant in pregnant women, and it is associated with later onset sepsis. All samples were positive for hyaluronidase and cytolysin/hemolysin, and had at least one Pilus Island; therefore clinical samples can be considered highly pathogenic. There were samples resistant to clindamycin, erythromycin and vancomycin; health professionals need to be aware of these results when prescribing antibiotics during labor. In conclusion, clinical isolates of GBS have a highly pathogenic and resistant profile, a worrisome scenario for pregnant women.Financing: FAPESP, CAPES

FCF034-2018CHARACTERIZATION OF VAGINAL MICROBIOTA AND QUANTIFICATION OF Lactobacillus sp. AND Streptococcus agalactiae DURING PREGNANCY

LUIZ GUSTAVO SPARVOLI (PG), RAMON VITOR CORTEZ DE GODOY (PG), CARLA TADDEI DE CASTRO NEVES

Department of Clinical and Toxicological Analysis, FCF/USP

Introduction and Objectives: The symbiosis between human and microorganisms has an important role in host health. In this sense, the gut is responsible for harbor the largest and most diverse microbial community in humans. Similarly, the vagina harbors a bacterial community of fundamental importance to women. These microorganisms act in the development and modulation of the immune system and compete for colonization sites, preventing pathogenic microorganisms’ colonization. The composition of the vaginal microbiota varies with age, pH, hormone secretion, menstrual cycle, contraceptive use and sexual activity. This study aimed to characterize and quantify the main bacterial species present in the vaginal microbiota of healthy pregnant and non - pregnant women through molecular biology techniques.Material and Methods: 41 healthy women were selected for the study, and vaginal samples from each participant were collected. The DNA of the samples was extracted and the DNA was submitted to sequencing of the 16S rRNA gene and quantified by the real time PCR technique. Participants selected were 25 pregnant and 16 non-pregnant women in reproductive age. Results and Conclusions: Firmicutes and Lactobacillus were found predominantly in both groups. However, an increase in relative abundance of Lactobacillus was found in the group of pregnant women. Also, in pregnant women, was observed a decrease of bacterial genera associated with a higher incidence of bacterial vaginosis. We believe that the action of gestation hormones positively influences the composition of the vaginal microbiota, thus guaranteeing the prevalence of bacterial genera responsible for maintaining a balance condition for a healthy genital tract. Financing: CNPQ


FCF035-2018DEVELOPMENT OF A FREE SOLUTION CAPILLARY ELECTROPHORESIS METHOD TO ASSAY AMINOPHYLLINE IN PHARMACEUTICALS

LARISSA CESAR FERNANDES DO NASCIMENTO (IC) *, FRANK ALONSO GAVILANO FAJARDO (M) *, CAROLINA COSTA PICOSSI (M) * , MARINA FRANCO MAGGI TAVARES**, MARIA INÊS ROCHA MIRITELLO SANTORO*, ERIKA ROSA MARIA KEDOR-HACKMANN* , MARÍA SEGUNDA AURORA-PRADO*

*Department of Pharmacy, School of Pharmaceutical Sciences, University of São Paulo. **Institute of Chemistry, University of São Paulo

Introduction and Objectives: Aminophylline is a 2:1 complex of theophylline and ethylenediamine. It is used to treat wheezing caused by asthma. The objective of the present work was to develop and validate a fast and economical method by free solution capillary electrophoresis (FSCE) to determine aminophylline in tablets. Material and Methods: The analyses were carried out in a Beckman and Coulter P/ACE MDQ capillary electrophoresis equipment equipped with a UV-VIS detector. A fused-silica capillary was used with an effective lenght of 20 cm x 75 µm i.d. The background electrolyte consisted of 20 mmol L -1 sodium tetraborate buffer solution, pH 9.2. The applied voltage was 17 kV, and the sample injection was performed hydrodynamically at 0.3 psi for 3s. The detection of the samples was set at 200 nm. Results and Conclusions: Good linearity (R2 > 0.99) over a concentration range from 56.00 µg mL-1 to 84.00 µg mL-1 was obtained; detection and quantitation limits were 5.71 µg mL-1 and 17.30 µg mL-1, respectively. The migration times for diphenhydramine (used as internal standard) and aminophyline were 0.5 min and 1.3 min respectively. The intra-day precision expressed as % RSD was less than 2%. The proposed FSCE method was found appropriate for determination of aminophylline in pharmaceutical formulations and could be successfully applied in quality control laboratories.Financing: FAPESP, CNPq.

FCF036-2018DEVELOPMENT AND CHARACTERIZATION OF EFAVIRENZ AMORPHOUS SOLID DISPERSIONS

*EDILSON MARCELO NAZARETH JÚNIOR (M), **FLÁVIO MACHADO DE SOUZA CARVALHO, *GABRIEL LIMA BARROS DE ARAÚJO, *CRISTINA HELENA DOS REIS SERRA

*Departament of Pharmacy, FCF/USP **Departament of Mineralogy and Geotectonics, IG/USP

Introduction and Objectives: Efavirenz (EFV) is an antiretroviral non-nucleoside reverse transcriptase inhibitor (NNRTI) part of the first-line treatment of HIV-1 infection. This compound belongs to class II of the biopharmaceutical classification system, i.e, it exhibits high permeability and low solubility (less than 10 mg/L). Due to this characteristic, the dissolution and, consequently, its bioavailability may be impaired, compromising the therapeutic efficacy of the medicine. In this study, EFV solid dispersions (SD) were obtained by solvent evaporation method aiming to enhance its solubility through amorphization. Material and Methods: During the SD development stage, a fast screening was performed to identify the best polymeric carriers to produce stable amorphous EFV dispersions. At this stage, SD were prepared on glass slides containing drug:carrier (PVP K-28/32, copovidone, poloxamer 188, poloxamer 407, HPMCP-55, HPMCP-55s and HPMCP-50) proportion of 1:3. The slides were submitted to stability study under accelerated conditions (40°C/75%) and the occurrence of crystallization was monitored by x-ray diffraction. The analyzes were conducted at the following time intervals: initial (T0), after 7 days (T1) and after 30 days (T2). Results and Conclusions: Results have shown that carriers poloxamer 188 and 407 were unable to promote the amorphization of SD and there are significant evidences of changes in crystalline structure during process of recrystallization. Financing: CAPES


FCF037-2018MODULATION OF THE FORMATION OF VOLATILE COMPOUNDS DURING RIPENING OF TOMATO (SOLANUM LYCOPERSICUM) CV. MICRO-TOM UNDER EFFECTS OF ETHYLENE AND AUXIN HORMONES

ELIS SILVA DE LIMA (IC), ERIC DE CASTRO TOBARUELA (D), EDUARDO PURGATTO

Department of Food Science and Experimental Nutrition, FCF/USP

Introduction and Objectives: Tomato fruit ripening is controlled by ethylene, while auxin is retarding the fruit ripening. The objective of this study was to evaluate the influence of hormones indolyl-3-acetic acid and ethylene on the volatile compounds in tomatoes (Solanum lycopersicum) ripening.Material and Methods: The fruits were divided into four groups: treated with ethylene (ET 100 ppm), indolyl-3-acetic acid (AIA, 100 uM), ethylene (100 ppm) followed by AIA (100 uM, ET+AIA) and the control group (C), without treatment. Color changes, loss of mass and ethylene production were evaluated on daily basis. The profiles of volatiles were analyzed in fruit at breaker and red stages. Results and Conclusions: In group ET, the typical increase in ethylene production and color shift were accelerated relatively to the control group, while treatments with AIA and ET+AIA delayed these processes. Relative to the control group, in breaker stage, treatment with AIA did not alter the pattern of volatile production, unlike fruits treated with ET. The group ET+AIA had a different profile compared to the other groups. Im red stage, there were slight differences between the groups, except the AIA and ET, which clearly differed from the control group, specially for some compounds such as hexanal. In breaker stage it was not observed a significant variation of transcripts associated to the volatile formation in ET and ET+AIA, such as SlLOX, while there was slight increase in AIA group. In red, there was a reduction of the levels of these same transcripts in ET and ET+AIA. Results indicate that both hormones, ethylene and auxin are capable of influencing the volatile profile. Financing: CNPq – PIBIC

FCF038-2018MICROREACTOR IN THE SYNTHESIS OF 2,4-THIAZOLIDINEDIONE DERIVATIVES WITH NITRATED ARYL ALDEHYDE

EDSON NASCIMENTO DOS SANTOS JUNIOR (PG), RENAN RODRIGUES DE OLIVEIRA SILVA (PG), MAURI SERGIO ALVES PALMA

Department of Biochemical and Pharmaceutical Technology - FCF/USP

Introduction and Objectives: Organic syntheses are traditionally carried out in multi-step batch processes. Presently, the need to develop more efficient, compact and sustainable processes has arisen. To meet this needs, several technologies were developed like miniaturization of equipment and the use of continuous flow. The main advantage of this type of process is the facility to scale up the process. In this context the microreactors appeared, being able to provide higher yields, generating less waste, and presenting higher heat and mass transfer than the usual batch reactors. This work utilized microreactors for the reaction of thiazolidine-2,4-dione (TZD) with 2-nitrobenzaldehyde, 3-nitrobenzaldehyde and 4-nitrobenzaldehyde.Material and Methods: Two solutions were prepared: a) 8 mmol of TZD, 4 mmol of pyrrolidine in 60 mL of methanol; b) 8 mmol of aldehyde in 60 mL of methanol. The continuous flow process was performed for several mean residence times, ?, and temperatures. Samples were collected and analyzed via HPLC-UV to determine the product yield. Results and Conclusions: This work showed that the use of microreactors is feasible for the synthesis of pharmaceutical intermediates. It was possible to reach yields higher than 70% for the reaction with 3-nitrobenzaldehyde at T = 120°C and time = 20 min. In addition, the thermodynamic parameters of the reaction and its kinetics were determined. The reaction with 2-nitrobenzaldehyde did not generate product, because the mechanism of reaction is different from the other two aldehydes. The 3-nitrobenzaldehyde was more reactive than 4-nitrobenzaldehyde, probably because it has less steric hindrance.Financing: FAPESP; CAPES.


FCF039-2018PHENOLIC JABOTICABA EXTRACT INHIBITS PANCREATIC LIPASE IN BOTH IN VITRO AND IN VIVO ASSAYS

DENISE MATOS DE OLIVEIRA (IC), MÁRCIO HÉRCULES CALDAS MOURA (PG), MARIA INÊS GENOVESE


Introduction and Objectives: Jabuticaba is a Brazilian native fruit rich in ellagitannins and anthocyanins, widely associated with beneficial effects on health. The objective of this study was to evaluate phenolic compounds from Sabara jaboticaba in form of a phenolic extract (PESJ), to inhibit pancreatic lipase in vitro and in vivo in animals previously induced to obesity by high-fat-sucrose diet (HFS). Material and Methods: The in vitro inhibitory capacity of the PESJ was assessed by fluorometric method according to Buchholz and Melzig (2016) using orlistat as control. The in vivo inhibitory capacity was assessed according to the following protocol: seventy-two male 8-wk-old C57BL/6J mice were randomly distributed into two groups: negative (Ch, 20 animals) and positive (HF, 52 animals) control group. For 14 weeks the Ch group was fed with standard diet for mice and the HF group was fed with a HFS diet. After this step, 10 animals of each group were properly euthanized and the white adipose tissues (WAT) were collected. The animals were redistributed into four groups and a new protocol was carried out for 14 more weeks: the Ch group was fed with the standard diet and water, the HF group was fed whit a the HFS diet and water, the J50 and J100 groups were fed with the HFS diet and the PESJ, obtained by solid phase extraction with octadecylsilane, at the dose of 50 and 100 mg gallic acid equivalent/kg body weight respectively. Results and Conclusions: The PEJS efficiently inhibited lipase pancreatic in both in vitro and in vivo assays. In the in vitro assay, its IC50 was one-third times lower than orlistat (PEJS IC50 271±15µg/mL, Orlistat IC50 401µg/mL). The supplemented groups J50 and J100 showed fecal lipid content 23% and 13,5% greater than HF group respectively. Financing: CNPq

FCF040-2018STABILITY-INDICATIVE METHOD FOR ANALYSIS OF RUPATADINE AND ITS DEGRADATION PRODUCTS USING HPLC

ANA CLAUDIA DE OLIVEIRA KITIZABOLO (IC), ANIL KUMAR SINGH

Department of Pharmacy, FBF/USP

Introduction and Objectives: Rupatadine is an antihistaminic drug, often used in the treatment of allergic disorders. To assure the efficacy and safety of this drug, it’s necessary to develop reliable and reproducible stability-indicative methods. The objective of present study was to develop a HPLC method for separation of rupatadine and its forced degradation products. Material and Methods: The method was developed on a Shimadzu® HPLC system, coupled to PDA-UV detector. All separations were obtained using a Luna C-18 (150x4.6mm) column. The rupatadine tablet samples were submitted to stressed conditions at elevated temperature (60°C), in neutral, acidic (HCl 1N), basic (NaOH 1N) and oxidative (H2O2 3%) medium, for 6 hours period.Results and Conclusions: The rupatadine and its degradation products were separated in reverse phase mode, with mobile phase constituted of acetonitrile and water, pH 3.0, a flow rate of 1.0 mL/min. The gradient was from 5 to 10% of ACN in 30 min and 50% in 45 min. The degradation was extensive in alkaline, oxidative and neutral conditions, after 6 hours. Rupatadine was more resistant to acidic medium. The rupatadine peak eluted at 38.8 min, with additional inherent impurity at 48.5min. In oxidative degradation, two additional peaks were observed at 36.6 and 47.2min, with λmax at 266 and 334 nm, respectively. The acidic stress presented a unique peak at 37.9min with λmax 272nm. A rapid degradation was observed in alkaline medium (1hr), with additional peak at 39.3min with λmax 236nm. No additional peak was observed in neutral hydrolysis, probably due to lack of chromophores. The obtained result may contribute in the stability assessment of rupatadine. Financing: Acknowledgements: Undergraduate Pro-Rectory of USP & FAPESP


FCF041-2018CRYSTALLIZATION STUDIES ON CIPROFIBRATE

CRISTIANO DE CARVALHO LEÃO* (IC), ILIA ALEKSEEVICH REPIN* (M), HUMBERTO GOMES FERRAZ*, FABIO FURLAN FERREIRA**, GABRIEL LIMA BARROS DE ARAUJO*

*Departamento de Farmácia, Faculdade de Ciências Farmacêuticas, Universidade de São Paulo **Centro de Ciências Naturais e Humanas (CCNH), Universidade Federal do ABC

Introduction and Objectives: Induction time is the period of time between the moment that supersaturation is achieved and the point at which the first crystals are detected. In this context, the application of methods that explore the stochastic nature of crystallization combined with different nucleation kinetics of crystal seeds allows charting crystal growth variations with different solvents and saturations that can lead to findings during the evaluation of the polymorphism phenomenon in organic crystals, especially in drugs. The objective of this study is to identify the induction times of Ciprofibrate (CIP) by measuring its crystal nucleation rates from selected solvents under different supersaturation conditions Material and Methods: Ciprofibrate (2- (4- (2,2-dichlorocyclopropyl) phenoxy) -2-methylpropanoic acid) acts by binding and activating nuclear receptors called “perxissome proliferator-activated receptors” or PPARs, increase lipid oxidation and lipase gene expression. Crystal16 allows to identify crystal nucleation rates of solutes mixed with selected solvents under different supersaturation conditions and temperatues by providing a controlled environment of cycling temperature, monitored by transmissivity of light during a determined period. Measurements were performed of Ciprofibrate in Toluene in 6 cicles of of 3 hours of heating and quenching. Results and Conclusions: The preliminary results show that crystallization of ciprofibrate in toluene yields relatively short and consistent induction times (10 minutes or less) at same supersaturation ratios. Further statistical treatment of the measurements shall be performed, and experiments replicated.Financing:

FCF042-2018VOLTAMMETRIC BEHAVIOR STUDY OF NITROFURAZONE AT MULTI-WALL CARBON NANOTUBE-MODIFIED GLASSY CARBON ELECTRODE IN AQUEOUS MEDIA

*CHARLES DE LIMA BRITO (PG), *ELIZABETH IGNE FERREIRA, **MAURO AQUILES LA-SCALEA

*Departamento de Farmácia, FBF/USP, **Departamento de Química, UNIFESP, Diadema

Introduction and Objectives: Carbon nanotubes, NTs, have been greatly prominent due to their varied electronic properties, which allow the transduction of signals. The NTs chemical functionalization improves detection traits, eliminating interferences from commercial NT. Based on the foregoing information, this work aims to evaluate the functionalization influence of carbon nanotubes on glassy carbon electrode by means of electrochemical reduction behavior of nitrofurazone applying cyclic voltammetry.Material and Methods: All voltammograms were recorded by using an electrochemical cell with three electrodes: Ag/ACl, Pt and working electrode. Three kinds of working electrodes, functionalized carbon nanotube (GCE-NTC-F), carbon nanotube (GCE-NTC-N) and GCE without modification were evaluated. The chemical functionalization of carbon nanotubes was carried out in a mixture of sulfuric and nitric acids. Solvents such as H2O, EtOH, MeOH, ACN, DMSO, DMF, 1,3-dioxolane were evaluated to get the best NTs dispersing agents. Three microliters of the suspensions were used and dried at room temperatureResults and Conclusions: The 1,3-dioxolane solvent was shown to be the best, as it presented greater current signals, despite being nontoxic.It can be observed that there are no significant differences among the potential values obtained. The current peak values of both modified electrodes at pH 4.02 are similar and, at pH 7.41, GCE-NTC-N presented the highest Ip,c value, indicating that the modification influences positively the electrochemical response of the hydroxylamine derivative formation from the nitrofurazone reduction. Moreover, the functionalization contributes actively to the elimination of the “ghost peaks”, which are irreproducible register by GCE-N. Financing:


FCF043-2018PRE-CLINICAL SAFETY OF POLYMERIC NANOSTRUCTURES WITH ENCAPSULATED DNA REPAIR ENZYMES

CAMILA FORSTER (IC), **ADRIANO MARIM DE OLIVEIRA (D), *CAMILA AREIAS DE OLIVEIRA (D), *CARLOTA DE OLIVEIRA RANGEL-YAGUI (D)

*Department of Biochemical and Pharmaceutical Technology - University of São Paulo ** Bionanomanufacture Nucleus - Institute for Technological Research – IPT, São Paulo

Introduction and Objectives: Skin cancer is one of the most occurring types of cancer due to the accumulation of free radicals and reactive oxygen species induced by UV radiation. In this project, we investigated the pre-clinical security of Pluronic L121 catalase-loaded polymersomes (PO) for topical application. Catalase (CAT) is a protein of the skin’s biological antioxidative system that removes hydrogen peroxide, by converting it to water and oxygen. Material and Methods: The nanostructure was developed by spontaneous formation with CAT 0,24 mg/mL, 0.2% w/v of Pluronic L121, stirring speed of 750 rpm and 48h speed time. In vitro cytotoxicity and phototoxicity was carried using two cell lineages, SIRC and NCTC929, and HET-CAM method.Results and Conclusions: It was verified that CAT was non-irritant whereas the PO decreased cell viability in a concentration-dependent manner (IC50: SIRC= 49,6 ± 13,5 µg/mL; 929= 83,3 ± 8,3 µg/mL), being classified as slightly irritant, the result close to the positive control sodium dodecyl sulfate (IC50: SIRC= 56,6 ± 1,5 µg/mL; 929= 26,0 ± 4,2 µg/mL). Both were considered non- phototoxic. HET-CAM proved to be more adequate to predict the irritant potential of PO due to the chorioallantoic membrane being highly vascularized, as well as being a fast, inexpensive and sensitive in vitro method. We determined that CAT and the PO were non-irritant because they did not cause any disturbance to the membrane. Therefore, CAT-loaded polymeric vesicles are safe for topical application and could be seen as a promising active ingredient for active photoprotection. Financing: Fundação de Apoio ao Instituto de Pesquisas Tecnológicas (FIPT)

FCF044-2018PREPARATION, CHARACTERIZATION AND THERMOSTABILITY STUDY OF PEGYLATED CATALASE

BEATRIZ DE MENDONÇA ROCHA (IC)1; JOÃO HENRIQUE PICADO MADALENA SANTOS (M)1,2; GUSTAVO CARRETERO (M)3; CAMILA AREIAS OLIVEIRA (D)1; CARLOTA DE OLIVEIRA RANGEL-YAGUI1

1Department of Biochemical-Pharmaceutical Technology, FCF/USP; 2Department of Chemistry, University of Aveiro; 3Department of Chemistry, IQ/USP

Introduction and Objectives: Catalase (CAT) is an antioxidant enzyme that catalyses hydrogen peroxide decomposition. Recently, catalase has been suggested as potential molecule to treat vitiligo. For this reason, topical use of catalase in association with UV therapy has been suggested as a new treatment modality. Considering that, we investigated the PEGylation of catalase as a potential biobetter with enhanced long-term stability and increased bioavailability. Material and Methods: In order to define an optimal site-specific PEGylation protocol, different pH values were tested for PEGs with 10, 20 and 40 kDa. Purification of the PEGylated proteins was carried out by FPLC-SEC and the molecular weight of purified enzymes analysed by native electrophoresis. Protein structural studies and thermal stability assays were achieved through circular dichroism (CD) and the peroxidative activity was also measured in order to show the PEG attachment effect in catalase activity.Results and Conclusions: PEGylation reaction was more effective in pH 8,0, in which yields of 45%, 59% and 31% for CAT-PEG-10, 20 and 40 respectively were obtained. CD showed that PEG coupling did not compromise the secondary and tertiary structure of the catalase and thermostability assays showed that both CAT and the CAT-PEG variants exhibit a higher aptitude to resist to thermal denaturation. Finally, the peroxidative activity was also measured and PEG binding to the protein caused a considerable decrease in peroxidative activity, which was expected. These results allow a better understanding about PEGylation as potential tool in the field of biobetters. Financing: CAPES; CNPq; FAPESP


FCF045-2018SYNTHESIS OF AN INTERMEDIARY OF ROSIGLITAZONE

LEANDRO LEITE RECHE¹ (IC), RENAN RODRIGUES DE OLIVEIRA SILVA² (D), MAURI SÉRGIO ALVES PALMA²

¹Faculdades Oswaldo Cruz (FOC); ²Department of Biochemical-Pharmaceutical Technology, FCF/USP

Introduction and Objectives: Rosiglitazone is a drug with anti-glycemic properties used in the treatment of type 2 diabetes mellitus . Rosiglitazone belongs to the class of thiazolidinediones and binds to the PPAR? receptor of lipid cells, sensitizing the insulin action in humans. This gain in sensitivity allows the body to make a better use of the insulin produced. An alternative for intensification of processes is the use of flow microreactors. Compared to traditional reactors, the microreactor technology has the advantages of an excellent control of heat and mass transfer and more efficient homogenization. This work aims at the synthesis of the third step of the synthesis of Rosiglitazone for subsequent transposition for flow synthesis in capillary microreactors. Material and Methods: To a round bottom flask was added 2 mmol of thiazolidine-2,4-dione, 2 mmol of 4-(2-(methyl(pyridin-2-yl)amino)ethoxy)benzaldehyde and 0.8 mmol of pyrrolidine in 30 mL of ethanol. The mixture was refluxed for 2 h. At the end of the reaction, crystallization with glacial acetic acid and water at 4°C was performed. Subsequently, the product was recrystallized in ethanol.Results and Conclusions: After recrystallization the resulting product was analyzed on HPLC-MS. The major peak of the product was identified in the chromatogram, though it was impure. In the next steps, new methodologies will be tested to better purify the product, based on chromatographic methods. Samples will also be taken as a function of time to determine the reaction time in the batch process and then transpose the reaction for flow synthesis in the capillary microreactor, aiming to reduce the reaction time, obtaining higher yields and selectivity of the product. Financing: FAPESP.

FCF046-2018ANALYSIS OF NEW STRATEGIES FOR THE EVALUATION AND MONITORING OF THE UNDERGRADUATE TEACHING AT THE FCF/USP

MATHEUS DELAQUA ROCHA DE JESUS (IC); ROSARIO DOMINGUEZ CRESPO HIRATA

School of Pharmaceultical Sciences, University of São Paulo

Introduction and Objectives: The evaluation of the undergraduate courses at the USP has been limited to the low adherence of the students to the eletronic system SIGA, which is not linked to the Jupiterweb, a plataform for undergraduate courses. This study investigated new strategies to evaluate and follow-up the undergraduate course in Pharmacy of the School of Pharmaceutical Sciences.Material and Methods: Two strategies were used for evaluation of 25 disciplines of the Pharmacy course using printed and eletronic google-based forms, in 2016 and 2017, respectively. The adherence to these strategies and the scores of the disciplines were compared by statistical tests (chi-square and t-test). Results and Conclusions: Adherence to the print forms strategy ranged from 6.0% to 97.0% (mean=70.4%), whereas adherence to eletronic strategy ranged from 0.7% to 38.8% (mean=15.1%). The evaluation using printed forms had higher adherence in 23 disciplines (92.0%) compared to the eletronic forms (p<0.05). The mean scores of the disciplines ranged from 3.0±1.1 to 4.6±0.5 for printed forms and from 1.9±1.0 to 4.4±0.7. Printed forms showed higher scores in 16 (64.0%) discipines compared to eletronic forms (p<0.05). Seven disciplines (28.0%) had similar scores between printed and eletronic forms (p>0.05). It is likely that the adherence rate of the evaluation forms is not directly related to the disciplines scores. In conclusion, the printed form strategy has higher adherence than the eletronic forms probably because it is applied at a specific time in the classroom. Since the data analysis of the electronic forms is less time consuming, better communication strategies should be implemented to increase adherence of the students, which have important contribution for the improvement of the course.Financing: PUBE fellowship, USP


FCF047-2018CHITOSAN-DERIVATIVE FROM ANTARCTIC KRILL ( Euphausia superba ) AS A NEW FAT SUBSTITUTE ON MUFFINS

RAQUEL VALLERIO RIOS (D), RAQUEL GARZÓN*, CRISTINA M. ROSELL*, SUZANA CAETANO DA SILVA LANNES

Pharmaceutical-Biochemical Technology Dept, FCF/USP, São Paulo, Brazil *Instituto de Agroquímica y Tecnología de Alimentos, Valencia, Espanha

Introduction and Objectives: The role of fat substitutes, specially hydrocolloids-derived from animals in the food matrix, has been attributed to its structuring properties on cakes. In order to reduce the fat content on muffins formulations, a new ingredient (succinyl chitosan SC) was used as fat replacer.Material and Methods: Five formulations with different levels fat reduction (25, 50, 75, 100%) and control (0 %) were elaborated. Basic ingredients were used: wheat flour, sugar, whole dried egg, skimmed milk powder, baking powder and water. SC was added at the same amount for all formulations (2.0 g/100 g flour). The effects were evaluated for consistency (RVA) and density; muffin texture profile (TPA), color (L, a*, b*), volume, porosity, water activity (Aw) and sensory evaluation.Results and Conclusions: Viscosities of the batters were compared and the pasting properties showed similarities between the samples, with the exception of the breakdown parameter (816 mPa.s) which decreased when the fat was 75% reduced. Batter’s density did not present significant differences (P > 0.05) for all samples. The structure of the reduced-fat muffins containing SC was similar to the control, even SC improved the circularity of the gas cells (0.62 ± 0.04). On TPA properties, the presence of SC during storage of the muffins reduced the hardening rate (14 N/day) when SC was replaced up to 50%, although the drying rate was accelerated. 50 % of fat reduction decreased Aw (0.91 ± 0.01) compared to control (0.93± 0.01). On hedonic scale, the panels attributed the score 7 for muffins with 0 up to 50% of fat. Thus according to the results, SC has a great potential as a new fat substitute on muffins. Financing: CAPES; CNPQ

FCF048-2018PREDICTING ANTIMICROBIAL EFFECTIVENESS OF PRESERVATIVE SYSTEMS USING A FTIR CHEMOMETRIC METHOD AIMING QBD AND PAT APPLICATIONS

CAMILA FRANCINI FIDELIS PINTO (M), JULIANA THAYNARA FIDELIS PINTO (IC), FELIPE REBELLO LOURENÇO

Departamento de Farmácia, Faculdade de Ciências Farmacêuticas, Universidade de São Paulo

Introduction and Objectives: Recently, cosmetics and pharmaceutical products with minimal concentrations of parabens and other preservatives have a safety and commercial appeal, due to the controversy regarding the safety of preservatives. However, the use of preservative is essential to guarantee the microbial conservation of cosmetic and pharmaceutical products during their usage. Here, we developed a FTIR chemometric method to predict the antimicrobial effectiveness of preservative systems in pharmaceutical and cosmetic products, aiming Quality by Design (QbD) and Process Analytical Technology (PAT). Material and Methods: DoE approach was used to explain the antimicrobial effectiveness against Candida albicans (ATCC 10231), Escherichia coli (ATCC 8739), and Staphylococcus aureus (ATCC 6538), as functions of parabens concentrations. All 15 formulas were analyzed using a FTIR spectrophotometer equipped with ATR apparatus. Results and Conclusions: PLS regression models for the slopes of microbial death curves as functions of FTIR/ATR spectra were well adjusted, with R2 and R2- prediction of 0.993691 and 0.892115, 0.994730 and 0.87827, and 0.99568 and 0.92218 for Candida albicans, Escherichia coli, and Staphylococcus aureus, respectively. The proposed FTIR method applied into a PAT approach was able to predict the effectiveness of the preservative system in real time. This in silico prediction will allow a batch-to-batch control of the effectiveness of the preservative system, which resulted in a reduction in the risk of quality deviations, and consequently, would provide an increase in the efficacy and safety of the product.Financing: FAPESP - Fundação de Amparo à Pesquisa do Estado de São Paulo


FCF049-2018MODULATION OF EPIGENETIC MARKS AND TRANSFORMATION IN CULTURE OF NORMAL HUMAN CELLS SUPPLEMENTED WITH FUMARATE

JOSEANA DE OLIVEIRA*(M), EVERSON WILLIAN FIALHO CORDEIRO*(D), GIOVANNA CAROLINE SILVA*(IC), MATHEUS RELVAS DOS SANTOS*(IC); MARISA HELENA GENNARI DE MEDEIROS**; PAOLO DI MASCIO**; SILVYA STUCHI MARIA-ENGLER*; ANA PAULA DE MELO LOUREIRO*

*Department of Clinical and Toxicological Analyses, School of Pharmaceutical Sciences,USP.**Department of Biochemistry, Institute of Chemistry,USP

Introduction and Objectives: Fumarate is a metabolite that accumulates in cells deficient in fumarate hydratase (FH), which are prone to malignant transformation. However, little is known about the effects of its supplementation to normal cells that are not deficient in FH. We assessed if fumarate favors the transformation of normal cells. Material and Methods: Normal human bronchial epithelial cells (BEAS-2B) were exposed to three non-cytotoxic concentrations of fumarate (1, 5, 10 mM). Cell transformation was assessed by the soft-agar assay after 168 h of exposure. Colonies taken from soft-agar were cultured and, after 90 days, the cells were submitted to the invasion/migration assay in Boyden’s chamber. The levels of 5-methyl-deoxycytidine (5-mC) and 5-hydroxymethyl-deoxycytidine (5-hmC) in DNA were determined by HPLC-ESI-MS/MSResults and Conclusions: Fumarate increased the growth of colonies in soft-agar. The cells of the colonies due to the fumarate exposures presented, after 90 days in culture, greater migration/invasion capacity than those from the control group. The 5-mC levels were unchanged in cells exposed for 168 h to fumarate and in those grown in soft-agar. However, there was a decrease of the 5- hmC levels in the cells incubated with fumarate. Data indicate that fumarate at concentrations that can be found in the microenvironment of cells with inhibited FH may favor the transformation of surrounding normal cellsFinancing: FAPESP, CNPq, CEPID Redoxoma

FCF050-2018LIPOPHILIZATION AFFECTS THE ANTIOXIDANT CAPACITY OF SINAPIC ACID ALKYL ESTERS

LETÍCIA MAEDA CAJAIBA (IC)*, TAYSE FERREIRA FERREIRA DA SILVEIRA (PD)*, BRUNO BAREA**, PIERRE VILLENEUVE**, INAR CASTRO*

*Department of Food and Experimental Nutrition, Faculty of Pharmaceutical Sciences, University of Sao Paulo, Brazil **AMIS, SUPAGRO/INRA (CIRAD) - Montpellier, France

Introduction and Objectives: Phenolipids are obtained by a lipophlization reaction between saturated fatty alcohols (SFA) and phenolic compounds. They have been suggested to increase antioxidant protection of polyunsaturated fatty acids in food emulsions. Because antioxidant properties of phenolic compounds may be modified on becoming phenolipids, our aim was to evaluate how the antioxidant capacity (AOC) of sinapic acid was affected by lipophilization with different chain lengths SFA.Material and Methods: Sinapic acid was chemically lipophilized with C4, C6, C8, C10 and C12 SFA. Then, the AOC of free sinapic acid (FSA) and its alkyl esters was examined, at two concentration levels (50 and 100 µM). Results and Conclusions: Regardless the assay, chain length and concentration, all the phenolipids were found to maintain their antioxidant properties. However, in general, FSA exhibited higher AOC than all the phenolipids. Moreover, as expected, increasing antioxidant concentration improved AOC. Among sinapic acid phenolipids, antioxidant response varied greatly depending on the chain length and assay, showing no constant correlation between AOC and the chain length. This result may be partly attributed to polarity differences between the studied compounds and the solvents used as reaction media in the AOC assays. Globally, C12 and C10 phenolipids showed lower AOC than that of the other phenolipids by hydrophilic assays, and did not show a significant difference in lipophilic assays. Therefore, based on ours results, sinapic acid lipophilized with C4, C6 and C8 would be indicated for further studies in food emulsions.Financing: FAPESP


FCF051-2018EFFECT OF LONG-TERM CONSUMPTION OF PARTIALLY OXIDIZED N6 FATTY ACIDS ON ATHEROSCLEROSIS IN LDLR-/- MICE

THIAGO MOTA VIEIRA (IC), MARINA SAYURI NOGUEIRA (D), BIANCA SCOLARO (PD) AND INAR CASTRO

Department of Food and Experimental Nutrition, Faculty of Pharmaceutical Sciences, University of São Paulo, São Paulo, Brazil

Introduction and Objectives: The consumption of polyunsaturated fatty acids (PUFA) increased exponentially during the 20th century, and therefore, consumers are being chronically exposed to toxic compounds from the oxidation of these FA. Our aim was to show the effects of long-term consumption of oxidized N-6 FA on atherosclerosis in LDLr -/- mice. Material and Methods: Animals were divided into five groups. Three of them received a high fat diet with 20% of soybean oil in different oxidation levels: fresh (N6), consumption (N6-C), fried (N6-FR). The CONT+ group received a high fat diet with oxidized corn oil and CONT- group received a standard AIN-93M diet.Results and Conclusions: After 6 months we observed that total weight gain was greater in the N6 and N6-C groups, while the N6-FR group did not show the expected increase of body weight, despite the similar food intake. Although animals in the N6-FR group were leaner, plasma levels of total cholesterol, LDL and VLDL were increased. Relative liver weight was also increased in N6-FR group. The intake of high fat diet contributed to an increase in plasma levels of glucose in all groups, compared to control. No difference was observed in plasma concentration of C-reactive protein. Interestingly, none of the atherogenic diets altered the peroxidation status of the liver, as assessed by MDA quantification. In conclusion, we observed that weight gain was inversely proportional with the degree of oil oxidation and it was proportional to liver size-increase.Financing: FAPESP

FCF052-2018MEROPENEM IN PAEDIATRIC CRITICALLY ILL PATIENT UNDER RENAL REPLACEMENT THERAPY

LEONARD DE VINCI KANDA KUPA (PG)*, DAVID DE SOUZA GOMEZ**, ELSON MENDES DA SILVA JR**, JOAO MANOEL DA SILVA JR**, NILO JOSÉ DUARTE***, SILVIA REGINA CAVANI JORGE SANTOS*

*Clinical Pharmacokinetics Center, Pharmacy Department FCF USP **Burn Center, Division of Plastic Surgery and Burning ***Division of Central Laboratory HC FMUSP (Sao Paulo, SP - Brazil)

Introduction and Objectives: Meropenem is recommended to start antimicrobial therapy in ICU critically ill pediatrics with septic shock. Renal replacement therapy by continuous haemodialysis is currently applied based on degree of renal failure. Subject of study was to investigate if the empirical dose regimen can reach the therapeutic target against gram-negative strains MIC> 2mg/L in a septic burn pediatric patient submitted to continuous haemodialysis. Material and Methods: Protocol was approved by the ethical committee N.5484. HLF (M), 12yrs, 60kg, 155cm, total burn surface area (15%) plus inhalation injury was investigated. Meropenem was administered 16mg/kg q8h or q12h, extended 3hs pump infusion. Target was based on 60%fΔT>MIC, time to maintain bactericidal serum level. Only two blood samples were required (2 mL/each) at the end of infusion and two hours afterwards to estimate the predictive index of effectiveness. Drug serum measurements were done by liquid chromatography ultraviolet detection. Results and Conclusions: Important change on pharmacokinetics was found during septic shock impacting drug distribution. Target was attained against MIC 8mg/L strains during clinical course of septic shock therapy. We conclude that meropenem serum monitoring done in a real time permits an earlier clinical intervention and desired clinical outcome with cure of infection during antimicrobial therapy in ICU pediatric patient under renal replacement therapy. Financing: CAPES


FCF053-2018EVALUATION OF ANTIMICROBIAL ACTIVITY OF CASPOFUNGIN IN NOVEL NANODISPERIOSN FROM BIXIN

GLEICY SOUZA MORALES (IC), NATHALIA TERCILIA ARAÚJO (IC), IRENE SATIKO KIKUCHI

Departamento de Farmácia, FCF/USP

Introduction and Objectives: Bixin is the major component of Bixa orellana seed surface (urucum) and has been demonstrated to be an efficient drug carrier with almost absence of toxicity in mammalian cells. This compound can interact and disperse compound insoluble in water and also decrease the toxicity in mammalian cells. The aim of this project was to prepare a novel nanodispersion from bixin and caspofungin with low cost and to improve the efficacy to treat disease related to fungi as Candida sp. and decrease the toxic effects of active ingredient. Material and Methods: These preparations were evaluated by microbial assays and in comparison to commercial products. Results and Conclusions: Caspofungin at concentrations above 2.0 µM demonstrated to be efficient to inhibit Candida albicans ATCC 10231 viability and after 24 hours of interaction for all preparations tested. Studies with microbial growth inhibition zones formation were performed in solid culture media inoculated with same yeast and no difference was observed between samples with caspofungin in water and in nanodispersions of bixin by statistical analysis. In contrast to this, commercial products presented lower inhibition zone in relation to preparations with bixin and at both concentrations (3.0 and 6.0 µM). Then we concluded that nanodispersions of bixin did not demonstrate toxicity against Candida albicans and it allowed its applications as drug carrier. Preparations of caspofungin in nanodispersions of bixin demonstrated better results to kill C. albicans in comparison to commercial products at same concentrations. More assays and studies are necessary to understand better their mechanism of action and physico chemical properties. Financing:

FCF054-2018FLUCONAZOLE SERUM MONITORING IN PEDIATRIC BURN UNDER CONTINOUS HAEMODYALYSIS

CLAUDIAGARCIA MESSIANO(PG)*, LEONARD DE VINVI KANDA KUPA(PG)*,DAVID DE SOUZA GOMEZ**; ELSON MENDES DA SILVA JR**; JOAO MANOEL DA SILVA JR**; NILO JOSÉ DUARTE***; SILVIA REGINA CAVANI JORGE SANTOS*

*Clinical Pharmacokinetics Center Pharmacy Department FCF USP **Burn Center Division of Plastic Surgery and Burning ***Division of Central Laboratory HC FMUSP (Sao Paulo, SP - Brazil)

Introduction and Objectives: Fluconazole is recommended to combat Candida spp in ICU( Intensive Care Unit) critically ill pediatrics. Renal impairment currently occurs in septic patients; then the extracorporeal removing by renal replacement therapy must be applied based on renal dysfunction. Subject of study was to investigate if the empirical dose regimen can reach the therapeutic target against Candida spp species MIC ( Minimum Inhibitory Concentration) 2mg/L in a septic burn pediatric patient submitted to continuous hemodialysis. Material and Methods: Protocol was approved by the ethical committee N.5484. HLF (M), 12yrs, 60kg, 15% total burn surface area plus inhalation injury was investigated. Patient received fluconazole one hour pump infusion as follows: 400mg q12h on day 42 and 200mg q24h on day 50. Target was based on drug effectiveness index AUCss 0-24/MIC>25. Only two blood samples were required (2 mL/each) at the 3rd - 5th hr of started the infusion. Drug serum measurements were done by liquid chromatography. Results and Conclusions: Important changes on pharmacokinetics occurred with impact on drug effectiveness-safety; then a reduction on daily dose was required during therapy. Target was attained against MIC 4mg/L strains in both sets during clinical course of antifungal therapy. Fluconazol serum monitoring based on target attainment done in a real time permits an earlier intervention to reach the desired clinical outcome with cure of infection and safety in the pediatric patient under renal replacement therapy. Financing: CAPES


FCF055-2018MEROPENEM-VANCOMYCIN TO TREAT SEPTIC PAEDIATRIC CRITICALLY ILL PATIENT

THAIS VIEIRA CAMARGO (PG)*; ANA PAULA DE CARVALHO CANELA BALZI**; LUIZ MARCELO MALBOUISSON**; DAVID DE SOUZA GOMEZ***; NILO JOSÉ DUARTE****; SILVIA REGINA CAVANI JORGE SANTOS*

*Clinical Pharmacokinetics Center, Pharmacy Department FCF USP **Post Operative Unity ***Burn Center, Division of Plastic Surgery and Burning ****Division of Central Laboratory HC FMUSP (Sao Paulo, SP - Brazil)

Introduction and Objectives: Meropenem and vancomycin are currently combined to treat septic critically ill in the ICU (intensive care unit) caused by nosocomial strains. The aim of study was to investigate if the combined therapy can reach the therapeutic target against S. aureus, minimum inhitory concentration: MIC>1mg/L and P. aeruginosa MIC> 2mg/L in one pediatric burn. Material and Methods: Protocol was approved by the ethical committee N.5484. MCCV (F), 3yrs, 14kg, 109cm, total burn surface area (28%) with inhalation injury and renal function preserved was investigated. Meropenem was administered 3hrs pump infusion 20mg/kg trice daily and vancomycin 1hr infusion 10mg/kg 4 times a day. Only two blood samples were required (2 mL/each). Drug effectiveness is guaranteed by the time that drug level is maintained above the minimum inhibitory concentration (60%f?T>MIC) for meropenem, while the area under the curve/MIC ratio (AUCss

0-24/MIC>400) is applied to vancomycin. Drug serum measurements were done by liquid chromatography.Results and Conclusions: Important changes on pharmacokinetics of both antimicrobials were found during the clinical course of septic shock. Target was attained against MIC 8mg/L strains by meropenem and vancomycin effectiveness was reached only against MIC:1mg/L S. aureus during therapy. Drug serum monitoring based on pharmacokinetic-pharmacodynamic approach done in a real time permits a clinical intervention in a real time to reach the desired clinical outcome with cure of infection. Financing: CAPES

FCF056-2018EVALUATION OF Dunaliella Salina GROWTH AND CORRESPONDING ß-CAROTENE PRODUCTION IN TUBULAR PHOTOBIOREACTOR

ELEANE DE ALMEIDA CEZARE GOMES (PG)*, MARIA EDUARDA GONÇALVES LOUSADA (PG)*, MARINA ISHII*, ANIL KUMAR SINGH**, JOÃO CARLOS MONTEIRO DE CARVALHO

*Department of Biochemical and Pharmaceutical Technology, University of São Paulo, Brazil **Department of Pharmacy, University of São Paulo, Brazil

Introduction and Objectives: Fruits and vegetables are the main sources of carotenoids, but microalgae represent an alternative for the production of these natural pigments. The microalga Dunaliella salina may be applied for the production of ß-carotene. Cell growth of Dunaliella salina CCAP 19/18, and ß-carotene using different fresh medium replenishments (20% and 80%) in the tubular photobioreactor has been evaluated.IMaterial and Methods: The Dunaliella salina was inoculated into the reactor with an initial cell density of 2.5 x 105 cells mL-1. The cultivation was illuminated under a 12 h light/12 h dark cycle, with a light intensity of 50 µmol photons m-2 s-1, at 24 ± 1 °C, and pH 7.0 ± 0.2. The ß – carotene concentration was determined by HPLC Results and Conclusions: It was observed that, in a general way, the final concentration at the end of each cycle was about 5 x 105 cells mL-1. Cell productivities in the three cycles of production were 4.33 x 104 cells mL-1 day-1 and 3.36 x 104 cells mL-1 day-

1 with replacements of cultivation medium of 20% and 80%, respectively. These cell productivities are substantially higher than those obtained in batch process, what evidences the importance of the semi-continous process when is desired to avoid adaptation phase, commun when working with batch process. It was concluded that the carotenoid content with replacement of 20% of volume increased with the increase of the cycle (around 6%). Financing: CAPES


FCF057-2018DESIGN, SYNTHESIS AND BIOLOGICAL EVALUATION OF NEW SELECTIVE, NON-HIDROXAMATE MATRIX METALLOPROTEINASE INHIBITORS.

NUNO ALBUQUERQUE TAVARES FERREIRA DA SILVA (M), ROBERTO PARISE FILHO


Introduction and Objectives: Despite the failure in clinical trials from two generations of inhibitors (MMPI’s), the pharmacological inhibition of matrix metalloproteinases (MMP’s) is still broadly studied for cancer treatment. MMP-2 and MMP-9 inhibitors present high affinities to their targets due to the presence of a strong zinc-binding group in their composition. The hydroxamic acid is the most potent zinc-binding group known, however, it’s utilization leads to pharmacokinetic unstability and increased toxicity. The present work has defined as objectives, the design, synthesis and biological evaluation of new selective non-hydroxamate MMPI’s for MMP-2 and MMP-9. For the design, the analogues were planned using information from the binding mode of known selective MMPI’s, and other reported zinc-binding groups.Material and Methods: The synthetic pathway to the designed molecules is based on the literature. Only analytical grade-reagents and solvents were used in this study. The reactions were monitored by thin layer chromatography. The products were purified by column chromatography or recrystallization. The structural analysis was performed using 1H and 13C nuclear magnetic resonance (NMR) and LC/MS. The MMP-2 and -9 IC50 of the obtained compounds will be determined by zimography.Results and Conclusions: The four-step pathway to the phenyl series last intermediate was established with 40% global yield. The obtainment of the control-hydroxamate as final product was well succeeded with 45% isolated yield and it was fully characterized by NMR spectroscopy, along with the previous intermediates. The purity of the obtained product is to be determined. The formation of the other zinc binding groups and the other two series of analogues is the next step to work on.Financing: CAPES, FAPESP

FCF058-2018EVALUATION OF CELL PROLIFERATION AND APOPTOSIS IN MAMMARY TUMORS OF FEMALE OFFSPRING OF OBESE MALE MICE TREATED WITH OR WITHOUT ORANGE JUICE.

VANESSA CRISTINA DA SILVA (IC), LÍVIA BEATRIZ APARECIDA RIBEIRO SILVA (PG), THOMAS PRATES ONG


Introduction and Objectives: Breast cancer is the most common type of cancer among women in Brazil and in the world. More recently, it has been described that the paternal food consumption is related to the susceptibility of breast cancer. Therefore, this project had as objective to evaluate the effects of paternal obesity and orange juice consumption during preconception in the cellular proliferation and apoptosis of mammary tumors of female offspring. Material and Methods: Apoptosis of breast tumors of the female offspring (n=5/group) were quantified and the results were represented as mean number of apoptotic cells/1,000 cells per tumor. Cell proliferation was evaluated in breast tumor of female offspring (n=5/group) by Ki67 immunohistochemistry. Cell proliferation was quantified by assessing the number of Ki67 positive cells among 1,000 cells per tumor. The slides were evaluated using Image J software. Results and Conclusions: No differences (p>0.05) were observed between the groups CO, CS, OB and OS of female offspring regarding cell apoptosis and proliferation. These results indicate that paternal obesity and/or consumption of orange juice during preconception may not influence female offspring breast tumors in these parameters. Financing: CNPQ; Food Research Center (FORC).


FCF059-2018SYNTHESIS OF 4-(2-(METHYL(PYRIDIN-2-YL)AMINE)ETHOXY)BENZALDEHYDE

CAROLINE ARASKIRO PESSOA (IC), RENAN RODRIGUES DE OLIVEIRA SILVA (PG), MAURI SÉRGIO ALVES PALMA


Introduction and Objectives: Rosiglitazone is an antiglycemic drug of the glitazone class and it is used in the treatment of type 2 diabetes mellitus which works as an insulin sensitizer in humans. The drug connects to the PPAR&gamma in fat cells making them more sensitive to insulin, this means that the body makes a more efficient use of the insulin that it produces. Continuous flow microreactors have been applied in both, academic and industrial practices, as a method of process intensification due to its advantages over stirred tank reactors, such as excellent heat and mass transfer and more efficient homogenization. The objective of this work is to perform the synthesis of the second steo of the Rosiglitazone synthesis for subsequent transposition to continuous flow microreactors. Material and Methods: 6.57 mmol of 2-(methyl(pyridin-2-yl)amino)ethanol, 13 mmol of NaH in 12 mL of DMF was added in a round bottle flask. The mixture was stirred for 1h. 14.45 mmol of 4- fluorobenzaldehyde was added to the mixture and the reaction was maintained for 24h under nitrogen atmosphere. After the completion of the reaction, the mixture was poured into cold water (20mL) at 4°C, extracted with 100 mL of ethyl acetate, washed with 30 mL of brine and dried with sodium sulfate. The crude product was purified by column chromatography with mobile phase chloroform/methanol 10:1.Results and Conclusions: The yield of 52% by mass was obtained, and its identification was carried out by HPLC-MS (m/z) 257,31 (M+1). In the next step of this study, it will be taken samples along the reaction to determine the reaction time in batch process followed by transposition to continuous flow synthesis in capillary microreactor aiming to reduce the reaction time, improve yield and selectivity.Financing: FAPESP; PUB-PRG-USP

FCF060-2018DOXYCYCLNE OXIDATION WITH H2O2 CATALYZED WITH Fe2+: INFLUENCE OF H2O2 AND IONS Br-, I- AND Cl-

GABRIEL LIMA BRESSAN (IC), MAURI SERGIO ALVES PALMA


Introduction and Objectives: The influence of hydrogen peroxide and ions bromide, chloride and iodide, usual contaminants of industrial effluents, was verified on the degradation of Doxycycline by the Fenton process. Doxycycline is an important antibiotic and its presence in the effluents of pharmaceutical industries is in the order of hundreds of ppm. Material and Methods: In a glass flask protected from natural light in a thermostatic bath, 35°C, was transferred 200 mL Doxycycline solution 100 ppm under mechanical stirring. Then hydrogen peroxide and ferrous sulfate solutions were transferred to obtain the concentrations: 50, 100 and 150 ppm, hydrogen peroxide, and 5 ppm Fe2+ ions. Samples were collected and transferred to flasks containing Fenton inhibiting solution to inactivate the peroxide and precipitate de Fe2+ ion. The spectrophotometric method was utilized to determine the residual concentration of Doxycycline. It was also verified the influence of the ions bromide, iodide and chloride, in concentration of 100 ppm, maintaining constant the temperature and concentration of Fe2+. Results and Conclusions: The best result (98.1%) was obtained for 150 ppm hydrogen peroxide and 5 ppm Fe2+ in 60 min reaction time. For bromide ion the efficiency dropped to 61.6%. For chloride ion the efficiency dropped to 97.6%. For iodide ion the efficiency dropped to 14.3%. The low efficiency of the process in the presence of bromide and iodide ions may be due to the fact that these ions react significant with hydrogen peroxide. Thus we conclude that it is important to determine the nature and the concentration of anions present when the Fenton process is used. Financing: FAPESP.


FCF061-2018REACTION OF 4,6-DICHLOROPYRIMIDINE AND 4-METHOXYPHENOL IN BATCH PROCESS AIMING THE TRANSPOSITION TO CONTINUOUS FLOW IN MICROREACTOR

DIEGO DE OLIVEIRA SANTOS (IC); RENAN RODRIGUES DE OLIVEIRA SILVA (PG); MAURI SERGIO ALVES PALMA.

Department of Biochemical and Pharmaceutical Technology, FCF/USP Introduction and Objectives: Lobeglitazone is an oral medication used in the treatment of type 2 diabetes mellitus. Its pharmacophoric nucleus activates the PPAR? receptor, reducing the insulin resistance in the fat cell. Continuous flow microreactor is a device used in process intensification and allows ease of mixing, due to the small distances for diffusion, while high surface-to-volume ratio provides a better heat and mass transfer, improving the reactant conversion, product yield and selectivity, generating less waste, reducing the cost of effluent treatment for the industry. This work aims to perform the first step of the synthesis of Lobeglitazone in batch process for subsequent transposition to continuous flow in microreactor. Material and Methods: 6.71 mmol (1.00 eq.) of 4,6-dichloropyrimidine and 4-methoxyphenol, 13.4 mmol of NaH (2.00 eq) and 30 mL of anhydrous DMF was added in a round bottle flask under N2 atmosphere at room temperature. After 3 h and constant stirring, the mixture was poured into water at 4°C and extracted with 100 mL of ethyl acetate. The organic phase was washed with distilled water, brine and dried over sodium sulfate. The product was purified using flash chromatography (hexane/ethyl acetate 5:1). Results and Conclusions: 33% yield was obtained by mass and its identification was performed by HPLC-MS (m/z) 236.94 (M+1). In the next step, samples along the reaction will be taken to determine the reaction time as well as a screening of solvents in batch process for subsequent transposition to continuous flow synthesis in the microrreator, aiming to reduce the reaction time and improve thereaction yield. Financing: FAPESP, PUB-PRGUSP

FCF062-2018INFLUENCE OF CATION Fe2+ AND ANIONS NO3

-, CO32-, AND F- ON THE DEGRADATION OF DOXYCYCLINE BY

FENTON PROCESS

DANIEL HIDEKI NUNES SAITO (IC), MAURI SERGIO ALVES PALMA


Introduction and Objectives: This work aims to study the influence of nitrate, carbonate and fluoride anions on the degradation efficiency of Doxycycline, consumed in human medicine, by Fenton process in the absence of light. This process is based on the oxidation by hydroxyl radical generated by the decomposition of hydrogen peroxide in the presence of catalytic amounts of Fe2+ ion. Doxycycline is present in effluent from pharmaceutical industries and nitrate, carbonate and fluoride anions are usual effluent contaminants. Material and Methods: The reaction medium was prepared with 100 ppm doxycycline solution in a glass flask isolated from natural light, Fe2+ (2.5, 5.0 and 10.0 ppm) and hydrogen peroxide (100 ppm). Doxycycline was analyzed spectrophotometrically based on the complex formation of the drug with sodium molybdate. The reaction solution was put in the thermostatic bath, 35°C, with mechanical stirring. Samples were taken as function of time. Tests were also performed to verify the influence of ions nitrate (5000 ppm), carbonate (50 ppm) and fluoride (100 ppm), keeping fixed the temperature, hydrogen peroxide and of Fe2+ concentration.Results and Conclusions: The best result of degradation was obtained for 100 ppm of hydrogen peroxide and Fe2+ 5 ppm with 97% efficiency, and reaction time 80 min. For the Fe2+ ion, the efficiency dropped to 93.6 and 89.4% for concentrations of 2.5 and 10 ppm, respectively. The nitrate ion did not influence the process. The ion carbonate and fluoride decreased the efficiency to 95.9%. and 76.4%, respectively. Thus we conclude that it is important to determine the nature and the concentration of anions present when the Fenton is used. Financing: FAPESP.


FCF063-2018EXPLORATORY DATA ANALYSIS OF A 5-NITRO FURFURILIDEN LIBRARY WITH ACTION AGAINST LEISHMANIA INFANTUM PROMASTIGOTES

CAROLINA ARRUDA BRAZ¹(D), EUZÉBIO GUIMARÃES BARBOSA², JOSÉ ANGELO LAULETTA LINDOSO³, LEOBERTO COSTA TAVARES¹

¹School of Pharmaceutical Sciences, USP. ²Federal University of Rio Grande do Norte, UFRN. ³Institute of Tropical Medicine, USP

Introduction and Objectives: Leishmaniasis, a neglected disease causes over 300 000 deaths/year. New treatments are needed since current therapy lead to severe side effects. N- alcylhydrazone library derivated from nifuroxazide was investigated due its potential activity against Leishmania infantum promastigotes. Previous results indicate the azometinic portion and 5-nitro furan as important scaffold for Kinetoplastid inhibition. This work aims to perform an exploratory data analysis in order to understand the molecular descriptors correlated to activity. Additionally the investigation provides useful SAR regarding the optimization of activity inhibiting profile. Material and Methods: Topological, constitutional, steric, electronic, thermodynamic and hydrophobic descriptors were calculated. A matrix of 41 rows (40 compounds and nifuroxazide) and 57 independent variables were analyzed through multivariate approaches HCA/PCA. Descriptors not lineally correlated to the biological activity - pIC50 (cut-off R = 0.2) and ill spread were removed. HCA and PCA loading plots were conducted with Pirrouette 3.11. Results and Conclusions: Dendogram presented 4 clusters sorted according to substituent nature, where active compounds grouped in cluster with hydrophobic substituents. PCA findings show 3 principal factors (PC) discriminating 86% total variance from original data (49.4% - PC1; 20.5% - PC2; 16.2% - PC3). PC1 loadings values were higher for topological/ constitutional (Platt, Wiener, RB) descriptors; hydrophobic (ClogPWM, PSA) for PC2 and PC3. PCA/HCA show complementary results where bulkier and hydrophobic substituents show higher activity against leishmania. Financing: CAPES

FCF064-2018USE OF BASES PIPERIDINE, PYRROLIDINE, 1,2-DIAMINOETHANE, MONOETILAMINE AND 2-METHYLAMINOETHANOL IN THE SYNTHESIS OF (Z) -5- (4-CHLOROBENZYLIDENE) THIAZOLIDINE-2,4-DIONA

MARCIO JOSÉ DE AGUIAR (M), LEANDRO RECHE LEITE (IC), MAURI SERGIO ALVES PALMA


Introduction and Objectives: Thiazolidine-2,4-dione (TZD) derivatives or glitazones are important allies in the treatment of type II diabetes mellitus. Derivatives such as (Z)-5- (4-chlorobenzylidene) thiazolidine-2,4-dione are used as building blocks in the construction of new drugs of this therapeutic group. The world diabetes cases reach 382 million people and should reach 471 million in 2035. Due to the restriction of the commercialization of piperidine, the usual promoter base of the reaction, the objective of this work was to seek new bases as an alternative for the synthesis of the product of interest.Material and Methods: Into a three-necked round bottom flask was added 4mmol p-chlorobenzaldehyde, 4 mmol TZD in 60 mL ethanol under heating. After reaching reflux, the reaction promoter base was added. The tested bases were: piperidine, pyrrolidine, 1,2-diaminoethane, monoethylamine and 2-methylaminoethanol in the amounts 3.2; 2,4; 3.0; 3.2 and 3.2 mmol. Samples were taken as a function of time to determine reaction time and product yield. Samples were analyzed on HPLC-UV.Results and Conclusions: The results showed that the yield of the reaction with piperidine was 67% in 480 min, pyrrolidine 98% in 80 min, 1,2-diaminoethane 43% in 130 min, monoethylamine 37% in 130 min and 2-methylaminoethanol 17% in 130 min. Compared with the yield obtained with piperidine, the base which showed the best result in relation to reaction time and yield was pyrrolidine, so it proved to be the best substitute. Financing: FAPESP; CAPES


FCF065-2018TECHNOLOGICAL DEVELOPMENT OF PROBIOTIC SUPPLEMENT FOR ZOOTECHNICAL IMPROVEMENT OF BROILERS

MAGALI T. UONO(M), SIBYLLE S. HACKER(D), CATARINA V. MANFRINATO(IC), MARIANA M. MATSUO(IC), SVETOSLAV D. TODOROV*, CRISTINA S. B. BOGSAN

FCF USP Depto. de Tecnologia Bioquímico Farmacêutica, *Depto. de Alimentos e Nutrição Experimental

Introduction and Objectives: The probiotic microorganisms Bacillus spp. are attractive for the inherent stability due to their characteristic of spore-forming bacteria. Spores allow prolonged shelf life and increase the ability to survive gastric barriers. Two bacteria, BVB1 and BVB5, were studied in order to identify which strain would have the best technological performance for the development of a novel functional food with high performance in the zootechnical improvement of broilers. The characterization was performed according to the protocol which included phenotypic and genotypic tests for determining the probiotic potential of the strains. The results of the tests revealed that BVB1 possesses the best technological characteristics and can be used with efficiency in the improvement of the zootechnical performance of broilers. Material and Methods: Observations of colonies and their smears staining by Gram were performed for the Morphological Test; catalase test, Hydrolysis of starch, Egg-yolk lecithinase, growth in NaCl at 6.5%, Hydrolysis of gelatin, Motility, Formation of indole, Voges-Proskauer, Nitrate reduced to nitrite, were conducted for the Biochemical Tests; MALDI-TOF/MS was performed for the classification based on characteristics of protein expression patterns; Enzymatic assays of amylase, protease and lipase of BVB1 and BVB5 were performed; and Sequencing of full genome of BVB1, were the adopted methods.Results and Conclusions: The results of the tests revealed that BVB1 possesses characteristics of high probiotic potential and can be used with efficiency in the improvement of the zootechnical performance of broilers. Further studies are needed to ensure the safety of this strain. Financing: Capes

FCF066-2018PHASE DIAGRAMS OF PEG/SALT AND PEG/IONIC LIQUIDS FOR THE PURIFICATION OF PROTEIN DRUGS

AMANDA ROBERTA PEREIRA DA SILVA (M) ELOÍSA GUEDES CAVICHON (M) JOÃO HENRIQUE PICADO MADALENA SANTOS (D) CARLOTA OLIVEIRA RANGEL YAGUI


Introduction and Objectives: Aqueous biphasic systems (ABS) are biocompatible alternatives for traditional organic-water solvent extraction that can be used for drugs, especially protein drugs. These systems are mostly composed of water with no volatile organic compounds, turning this purification platform mild for proteins and environmental friendly. ABS are formed by a combination of polymers, copolymers, salts or ionic liquids (ILs), and the most common polymer is poly(ethylene glycol) (PEG). PEG is also used to lower the immunogenicity and increase plasma half-life of protein drugs, by means of a covalent attachment known as PEGylation. Material and Methods: Was mapped out the phase diagrams of PEG/citrate and PEG/citrate + imidazolium-based ionic liquids ABS that will be further used for PEGylated proteins purification. The ILs used were 1-ethyl-3-methylimidazolium triflate (MIMTF) and 1-ethyl-3-methylimidazolium dimethylphosphate (MIMDMP) acting as adjuvants in PEG/citrate systems. Phase diagrams were obtained by cloud point titration and our results show that the molecular weight of PEG have a strong effect in the ability to form a two phase system, with the following trend: PEG 20000 > PEG 10000 > PEG 2000. The effect of PEG methylation in ABS formation was also evaluated since PEGylation is performed with methylPEG. Results and Conclusions: The presence of methyl group does not affect the ability to form two phases. The stability of a model protein L-asparaginase in the IL was investigated and we show that protein preserved activity in up to 2.5% of MIMDMP and up to 10% MIMTF. The purification of PEGylated proteins through ABS platforms is under investigation for the separation of unreacted proteins from the PEGylated ones. Financing: CAPES;CNPq; FAPESP


FCF067-2018POTENTIAL CANDIDATES FOR NEW ANTICANCER AGENTS: SYNTHESIS AND EVALUATION OF ANTITUMOR ACTIVITY OF TIOUREIDICS AND UREIDICS ANALOGS OF CAPSAICIN

GUSTAVO JOSÉ VASCO PEREIRA (M), ROBERTO PARISE FILHO

FBF – FCF/USP

Introduction and Objectives: The malignant neoplasias, diseases worldwide known as cancer, have one of the most expensive, toxic and low selective treatments in the current therapy. In addition, the continuous increase in the disease incidence also represents a major problem. Natural products are presented as alternatives many diseases, including cancer. Capsaicin, a natural product derived from Capsicum peppers, has antineoplastic properties, and therefore, can be used as a prototype to obtain analogues. Material and Methods: Four series were planned and with ureídics and thioureídics compounds. The synthetic strategy was based on the reaction of primary amines with isocyanates or isothiocyanates, attached to aromatic or alkyl substituents. Results and Conclusions: Twenty eight capsaicinoid analogues were designed based on molecular modifications such as bioisosterism, ring closure, addition of volumous functional groups and homology of methylenic chains. The modifications led to the synthesis of vanillic and piperonylic compounds, both with urea and thiourea as functional groups. It were performed a nucleophilic addition of primary amines (piperonylamine and vanillylamine) onto the carbonyl or thiocarbonyl of isocyanates and thioisocyanates. Characterization were elucidated using 1H and 13C NMR and melting point. Yields varied between 20% to 90%, so they were considered to be low to excellent. All compounds were evaluated in healthy and neoplastic cell cultures and evaluation of cell viability by MTT reduction. The most active compound presented a IC50 value of 55 µM in A2058 cells from human melanoma. A struture-activity relashionship study is been conducted, using the calculated properties of the analogues.Financing: Fapesp; Capes

FCF068-2018ELUCIDATION OF THE REACTION MECHANISM OF 2-(METHYL(PYRIDIN-2-YL)AMINE)ETHANOL SYNTHESIS USING THEORETICAL CALCULATIONS WITH DENSITY FUNCTIONAL THEORY (DFT)

RENAN RODRIGUES DE OLIVEIRA SILVA* (PG), PATRICK RODRIGUES BATISTA** (PG), LUCAS COLUCCI DUCATI**, MAURI SERGIO ALVES PALMA*

*Department of Biochemical and Pharmaceutical Technology - FCF/USP **University of São Paulo, Chemical Institute

Introduction and Objectives: In recent years, computational chemistry has made an interface between theoretical and experimental chemistry. DFT provides information to understand and elucidate reaction mechanisms. This work aims to describe the reaction mechanisms, kinetic and thermodynamic parameters of the synthesis of 2-(methyl(pyridin-2-yl)amine)ethanol, an intermediate of Rosiglitazone, a drug used in the treatment of type 2 diabetes mellitus. Material and Methods: Geometry optimization of all reactants, intermediates and product were performed with hybrid density function M062-X with base 6-311++g(d,p) in gas phase. Frequency calculation were carried out to obtain thermodynamics properties and to confirm all optimized structure provides only one imaginary frequency. All DFT calculations were carried out with Gaussian09.Results and Conclusions: Possible pathways of the mechanism occurs from the elimination of a chloride ion followed by a nucleophilic addition via a transition state of 2-(methylamine)ethanol and abstraction of the proton from the amine group by the chloride ion, forming the product and hydrochloric acid. The Gibbs energy of activation founded was 26.657 kcal/mol, which means the energy barrier for the transition state, while the barrier between the transition state and the product is 28.465 kcal/mol. The energy difference between product and reagent is -1.80 kcal/mol, showing a higher stability of the products against the reagents. As next step new hybrid density function and set of bases will be tested looking to evaluate which theoretical level is better suited to the proposed system. Financing: FAPESP


FCF069-2018SYNTHESIS OF 5-NITRO HETEROCYCLIC DERIVATIVES WITH ACTIVITY AGAINST Leishmania infantun

JÉSSICA GÜNTHER FANIZZI (IC), GLEN ELLA MEBIAME (IC), CAROLINA ARRUDA BRAZ (D), LEOBERTO COSTA TAVARES

Departamento Tecnologia Bioquímico Farmacêutica FCF/USP

Introduction and Objectives: Leishmaniasis is a neglected disease responsible for over 20.000 deaths/ year worldwide, affecting mainly developing countries. Aim of this work is synthesize 5-nitro-heterocyclic nifuroxazides derivates and evaluate their activity against promastigotes of L. infantum, denoting the search for alternatives to current treatment, which is complex and toxic to patients.Material and Methods: 5 compounds were obtained through three synthetic steps: esterification, hydrazinolysis and nucleofilic addition reactions followed up by recrystallization from DMF and acetonitrile.The structures were confirmed by H¹ and C¹³ NMR and purity was evaluated by melting point range. Log phase L. infantum promastigotes were maintained in M199 supplemented with 10% FCS and 2% filtered male urine at 25 ºC. Parasite viability was measured through MTT assay and IC50 was calculated for Nifuroxazide, Miltefosine and Amphotericin B.Results and Conclusions: The compounds were obtained from the synthesis: 4-penthyl-N’-[(5-nitrofuran-2-yl)methylene]benzohydrazide (C1): ?=40%; MP=232,7-235,4°C; 4- hepthyl-N’-[(5-nitrofuran-2-yl)methylene]benzohydrazide (C2): ?=68.2%; MP=179.7-181.0°C;4-hepthyloxy-N’-[(5-nitrofuran-2- yl)methylene]benzohydrazide (C3): ?=78.8%; MP=172.8-173.2°C; 4-biphenyl-N-(5-nitrofuran-2-yl)methylene]carbohydrazide (C4): ?=57%; MP=230.6- 235.0°C and 4-penpthyl-N’-[(5-nitrofuran-2-yl)methylene] cyclohexane carbohydrazide (C5): ?=84,86%; MP=194.8-194.9°C. Compounds 1-5 and nifuroxazide presented IC50 values of 1.55; 0.35; 0.22; 1.30; 1.65 and 5.93 µM. Miltefosine and amphotericin B had values of 8.58 and 0.03 µM. All compounds showed optimizing structures modifications in comparison to nifuroxazide with C3 presenting the higher activity inhibiting potential.Financing: CAPES, PUB/USP

FCF070-2018Physico-chemical characterization of kombucha

CAROLINY DE ALMEIDA SOUZA (M); CRISTINA STEWART BITTENCOURT BOGSAN

Tecnologia Bioquímico-Farmacêutica da FCF/USP

Introduction and Objectives: Kombucha is a traditional fermented drink from China, prepared by the fermentation of black tea sweetened with a mixed culture of bacteria and yeasts (SCOBY), which has aroused people’s interest in recent years due to its possible functional properties. The objective of this work is to perform the physical-chemical characterization of kombucha, the first necessary step to evaluate its possible functional capacity. Material and Methods: Carry out analyzes of titratable total acidity, pH (at temperatures 20ºC and 25ºC), density and dry residue. Results and Conclusions: It can be observed that kombucha has higher titratable acidity and pH and lower density and dry residue than black tea, and such values are enhanced according to the kombucha fermentation time (D1 to D14). Therefore, it is concluded that the fermentation of black tea promoted by SCOBY significantly modifies most of the analyzed parameters, revealing the importance of the physico-chemical characterization. This activity compose one of the steps of the research project on the evaluation of the possible anti-hyperglycemic capacity of kombucha in diabetic mice.Financing: Agência CAPES/Cota PROAP


FCF071-2018APPLICATION OF PHARMACOPEIAL METHODS TO ASSESS STABILITY OF PIOGLITAZONE HYDROCHLORIDE

HEITOR OLIVEIRA DE ALMEIDA LEITE (PG), ANIL KUMAR SINGH

Department of Pharmacy, FBF/USP

Introduction and Objectives: For qualitative and quantitative analysis of impurities and degradation products, stability indicative methods must be used. Pioglitazone HCl is an antihyperglycemic agent that acts primarily by reducing insulin resistance in the treatment of type 2 diabetes. The pharmacopeial methods often do not provide information regarding analysis of impurities and/or degradation products. The objective of this research was to access stability indicative capacity of pharmacopeial methods.Material and Methods:A Shimadzu® HPLC system, coupled to PDA-UV detector was used and separations were obtained with a C-18 (250x4.6mm) column. A factorial design (32) of the degradation mediums was used to evaluate impact of time-of-contact and variable intensity. The degradation medium factors considered in the design were, acid, alkaline, peroxide, and temperature/humidity besides photostability. The ability of the method to detect and quantify degradation products was assessed.Results and Conclusions: Pioglitazone HCl was found to be vulnerable to basic, oxidative, incident light and elevated temperature/humidity conditions. Acidic hydrolysis has no significant impact. The assay and impurity methods did not show mass balance, elucidating that the method is not stability indicative. In silico analysis, with the Zeneth® software, was used to establish correlations between the degradation process and the plausible degradation pathway and products. Thus, it is concluded that the data obtained in degradation studies, in this work, may serve as support in the stability studies, as well as subsidize to define transport and storage conditions for pioglitazone HCl. Financing: LIBBS Pharmaceuticals & FAPESP

FCF072-2018SYNTHESIS OF DENDRIMER PRODRUGS POTENTIALLY ACTIVE IN MALARIA AND TUBERCULOSIS

DIEGO CAMPOS PRIETO (M)1, SCOTT GRAYSON2, JEANINE GIAROLLA1

1Department of Pharmacy, Faculty of Pharmaceutical Sciences, University of São Paulo; 2Department of Chemistry, School of Science and Engineering, Tulane University

Introduction and Objectives: HIV/AIDS, tuberculosis, malaria represent a major concern in health in many regions of the world. The drugs available for treatment show several problems, such as toxicity and resistance against the parasite. Taking it into account, methods of molecular modification to improve pharmaceutical, pharmacokinetic and/or pharmacodynamic properties of bioactive compounds, such as prodrug design, is outstanding. Dendrimers have been attacking interest in biological applications and, considering the promising feature as drug carriers, the aim of this work is the synthesis of Bis-MPA dendrimer prodrugs (generations 0, 1 and 2) potentially active in malaria and tuberculosis.Material and Methods: The dendrimer functionalization is being performed through the coupling of succinic acid (as a spacer) and the drugs: primaquine, with antimalarial activity and isoniazid, which present action in the early stages of tuberculosis. Even now, the generation 0 of the Bis-MPA dendrimer containing primaquine (G0-Pq) and isoniazid (G0-Iso) were synthetized using Steglich esterification (EDC and DMAP).Results and Conclusions: The yields were 95% and 88%, respectively. The 1H,13C NMR and IR results indicated these products. Also, the generation 1 of Bis-MPA dendrimer functionalized with isoniazid (G1-Iso) was synthetized using the same method and the characterization using 1H,13C NMR and IR demonstrated the respective compound, with 33% yield. The perspectives include the analysis with MALDI-TOF mass spectrometer, as well as the synthesis of the generation 1 Bis-MPA dendrimer with primaquine and the generation 2 with both drugs. Also, we intend to evaluate the biological activities of the respective dendrimer prodrugs. Financing: CNPq


FCF073-2018OPTIMIZATION OF NEW DIHYDROFOLATE REDUCTASE INHIBITORS OF Mycobacterium tuberculosis (mtDHFR): MOLECULAR DOCKING, SYNTHESIS, EVALUATION OF ENZYME INHIBITION AND ANTIMYCOBACTERIAL ACTIVITY

ALFREDO DANILO FERREIRA DE SOUZA* (PG), JOÃO AUGUSTO RIBEIRO** (PG), GLAUCIO MONTEIRO RIBEIRO* (PG), GUSTAVO HENRIQUE GOULART TROSSINI*, MÁRCIO VINÍCIUS BERTACINE DIAS**, ROBERTO PARISE FILHO*

* Faculdade de Ciências Farmacêuticas da USP. Departamento de Fármacos e Medicamentos ** Instituto de Ciências Biomédicas da USP. Departamento de Microbiologia

Introduction and Objectives: Tuberculosis is considered an important infectious diseases. The relationship with HIV/AIDS, long treatment and multidrug resistance implies a need for new drugs. The mycobacteria enzyme dihydrofolate reductase (mtDHFR), is a poorly explored target and has great potential. Preliminary studies obtained small fragments with low profile affinity to mtDHFR, however with the potential to become lead compounds. Thereby, the object of this work is to design and synthesize fragments anologues and to evaluate their enzyme inhibition and antimycobacterial activity.Material and Methods: The compounds were designed using bioisosterism approach and information about ligands and the three-dimensional structure of mtDHFR. For the syntheses, alkylating and cycloaddition protocols and were carried out. The structure of products was elucidated with NMR spectrum of 1H and 13C. HPLC was used to evaluate the purity of compounds. Enzymatic inhibition assay was performed using fluorometric method. Results and Conclusions: Twenty two compounds were obtained with satisfactory yields. The chemical shifts are consistent with those described in literature and the purity of compounds was above 95%. Preliminary results of the enzymatic inhibition assay were consistent with that obtained in the docking assays. The compounds also were sent to the antimycobacterial assay and the results are being awaited.Financing: CNPq; CAPES; FAPESP

FCF074-2018DESIGN, SYNTHESIS AND ANTITUMOR EVALUATION OF SANGUINARINE ANALOGUES

THAIS BATISTA FERNANDES (D), ROBERTO PARISE FILHO

Departament of Pharmacy, FCF/USP

Introduction and Objectives: Bcl-xL is a pro-survival member of Bcl-2 protein family, which is overexpressed in tumor cells and related to drug resistance. This protein can be inhibited by sanguinarine, natural quaternary benzophenanthridine alkaloid that has been shown to be an interesting prototype for anticancer drug design. Thus, the aim of this work is optimizing the benzophenanthridine scaffold in an attempt to identify new molecular entities with antitumor potential.Material and Methods: Benzaldehydes were condensed with naphthylamine, followed by reduction with sodium borohydride to give secondary amines. Suzuki coupling was employed to provide isoquinoline analogues. These molecules were further reacted with methyl iodide to provide N-methylated compounds. The final products were purified by recrystallization and collumn chromatography and characterized by 1H and 13C NMR and melting point. Purity was verified by high performance liquid chromatography. Results and Conclusions: The synthesis and purification of twelve analogues were successful, with yields between 28 and 78 %. NMR data of final compounds corroborates the formation of molecules, which presented purity superior to 95 %. Compounds will be evaluated about their cytotoxicity and Bcl-xL inhibition and submitted to molecular modeling in order to obtain insights about their strutucture-activity relationship. It is expected that these compounds present better results than its prototype.Financing: CNPq; FAPESP.


FCF075-2018DIFFERENCES BETWEEN BUFFALO MILK MATRIX AND BOVINE MATRIX IN LIPID PROFILE

ALESSANDRA PRESTES (M), MARIANA MITIKO (IC), BRUNA YOSHIDA (IC), CRISTINA STEWART BOGSAN.

Department of Biochemical Pharmaceutical Technology, FCF/USP

Introduction and Objectives: Fermented milks are the main vehicle for the consumption of probiotic microorganisms. The milk matrix, in addition to its nutritional capacity, is rich in bioactive components, among them peptides and fatty acids, these components vary according to the animal species used, and buffalo milk has been distinguished by its nutritional properties as a high content amino acids, minerals and fatty acids (AG). Dairy derivatives from buffalo matrix present superior yield when compared to dairy derivatives from other matrices, such as cow.GAs have a potential contribution to the regulation of immune functions in humans, especially conjugated linoleic acid (CLA), which has an anti-inflammatory capacity, as well as inhibition of the carcinogenic process. Bifidobacteria are probiotic bacteria capable of modifying the concentration of these AG in the fermented matrix. The objective of the present work was to compare the lipid profile, especially CLA, in the fermentation of the bovine and buffalo dairy matrices by the strain Bifidobacterium animalis subs. lactis HN019 in the search for higher CLA content. Material and Methods: The methodology used was based on the ISO 14156 and ISO 15884 methods. The lipid profile in buffalo fermented milk is 1.85 times higher when compared to bovine milk, whereas the CLA content in fermented buffaloes represents only 1.25 % of the total fatty acids of the product while the bovine fermented milk has 4.64% of CLA. Results and Conclusions: Thus, buffalo fermented milk does not present the potential functional effect offered by CLA, and can not be characterized by this parameter as a functional food.Financing: capes

FCF076-2018INSULIN MODULATES BRONCHIAL HYPERRESPONSIVENESS IN LUNG ALLERGIC INFLAMMATION IN NON-DIABETIC AND DIABETIC MICE

SABRINA DE SOUZA FERREIRA (D), MARISTELA TSUJITA (PD), FERNANDA PEIXOTO BARBOSA NUNES (PD), FELIPE BECCARIA CASAGRANDE (D), JOILSON OLIVEIRA MARTINS

Department of Clinical and Toxicological Analyses, FCF/USP, São Paulo, Brazil

Introduction and Objectives: Experimental and clinical studies have indicated that the inflammatory response is impaired in diabetic patients. Objectives: The present study evaluated the modulatory role of insulin in late pulmonary allergic inflammation in non-diabetic and diabetic mice. Material and Methods: Diabetic male BALB/C mice (alloxan, 50 mg/Kg, i.v., 10 days; CEUA/FCF/USP-490) were sensitized by i.p. injection of ovalbumin (OA) in Al(OH)3 14 days before multiple OA challenges and insulin treatment. The following parameters were analyzed: a) bronchial hyperreactivity; b) inflammatory cell infiltration into the bronchoalveolar lavage fluid (BALF); c) cytokine concentration (IL-4,IL-5,IL-10,IL-13,TNF-a,IFN-?,VEGF,TGF-ß) in lung homogenate; d) immunoglobulins present in the animal serum (IgG1, IgG2a, IgE). Results and Conclusions: Compared to non-diabetic OVA mice, diabetic OVA mice had the following decrease: a) inflammatory infiltrate (2.7-fold) and absence of eosinophilia b) bronchial hyperresponsiveness (1.5-fold) c) the concentration of serum IgG1 (2-fold), IgE (1.6-fold), d) the concentration of BALF: IL-4 (3-fold), IL-5 (1.8-fold), IL-13 (4-fold), TNF-a (2-fold), VEGF (1.8-fold) and TGF-ß (1.7-fold). Treatment of diabetic mice with insulin completely restored cell migration, eosinophils population, concentration of IgG1 and IgE in the serum, concentration of IL-4, IL-5, IL- 13 and TNF-a in the BALF. These results suggest that insulin modulates bronchial hyperresponisiveness by regulating cell migration and cytokine concentrations in the allergic lung inflammation in non-diabetic and diabetic animals.Financing: Financial support: FAPESP (2017/11540-7); CAPES; CNPq (301617/2016-3).


FCF077-2018THE ROLE OF LEUKOTRIENES IN INFLAMMATION, INSULIN PATHWAY AND MACROPHAGE POLARIZATION IN TYPE 1 DIABETES

JOÃO PEDRO TÔRRES GUIMARÃES (D), LUCIANO RIBEIRO FILGUEIRAS, SONIA JANCAR, JOILSON OLIVEIRA MARTINS*

Department of Immunology, Institute of Biomedical Sciences, ICB/USP *Department of Clinical and Toxicological Analyses,FCF/USP

Introduction and Objectives: Type 1 diabetes (T1D) is a metabolic disease associated with systemic low grade inflammation and macrophage reprogramming. In T1D mice, systemic levels of leukotrienes (LTs) are elevated and might contribute to systemic low grade inflammation and insulin resistance in metabolic active tissues. In our study we investigated the role of leukotrienes (LTs) in muscles from T1D WT and knockout 5-lipoxigenase mice (5LO-/-) enzyme responsible for LTs synthesis.Material and Methods: T1D was induced in 129 sve mice (WT) and knockout of 5-lipoxigenase (5LO-/-) by streptozootocin (60 mg/kg, i.p). Response to insulin was evaluated by Insulin Tolerance Test (ITT), insulin concentration by ELISA and phosphorylation of Akt by Western Blotting. Gene expression of insulin receptor, macrophage phenotypic markers and interleukin 6 was evaluated by qPCR and cytokines by ELISA. Results and Conclusions: After a single dose of insulin, the reduction of glicemia was more pronounced in 5LO-/-T1D mice compared to WT. In muscles of T1D mice, expression of pro-inflammatory IL6 was higher in WT whereas IL10 level was higher in 5 LO-/- mice. Moreover, we found a higher expression of M2 markers Ym1 and Arg1 in the muscles of 5LO-/-T1D mice and higher pAkt. The expression of insulin receptor gene in T1D mice was higher in 5LO-/- compared to WT indicating a possible regulation in the expression of this gene mediated by LTs. These results suggest that LTs have impact on muscle-associated macrophages in mice with T1D and have impact on insulin receptor signaling pathway.Financing: FAPESP; CAPES; CNPq

FCF078-2018IMMUNOGENICITY OF A MULTI-ANTIGEN FORMULATION BASED ON Plasmodium vivax MALARIA VACCINE CANDIDATES

JANAÍNA TENÓRIO NOVAIS (IC), TARSILA MENDES DE CAMARGO (PD), RODOLFO FERREIRA MARQUES (D), KÁTIA SANCHES FRANÇOSO, IRENE DA SILVA SOARES

Department of Clinical and Toxicological Analyses, School of Pharmaceutical Sciences, University of São Paulo

Introduction and Objectives: Considering the complexity of malaria parasite life cycle, we have hypothesized that an effective vaccine formulation against Plasmodium vivax malaria should contain antigens expressed on different parasite stages. For this, we generated recombinant proteins based on the circumsporozoite protein (liver stage - PvCSP-AllCT) and apical membrane antigen 1 and merozoite surface protein 1 (blood stage - PvAMA1-MSP119). Material and Methods: The first construct contains the PvCSP conserved C-terminal region as well as its variant repeat domains (VK210, VK247, and P. vivax-like) in tandem. The second construct consists of a fusion between AMA1 and MSP1 immunodominant domains. Both recombinant proteins were expressed in Pichia pastoris yeast. In order to estimate their ability to induce antibodies, C57BL/6 mice were immunized with 10?μg of each protein alone or in combination (PvCSP-AllCT + PvAMA1-MSP119) in the presence of 50 μg of Poly (I:C) adjuvant. IgG and respective subtypes (IgG1, IgG2b, IgG2c and IgG3) were determined in serum samples by ELISA two weeks after the third dose.Results and Conclusions: After measuring the antigen-specific IgG and isotypes titers, it was clear that all formulations elicited a balanced IgG1/IgG2c ratio, suggesting a mixed Th1/Th2 response. However, the highest IgG1 and IgG2c titers against PvCSP variant repeats or PvAMA1 were identified in mice groups immunized with the protein combination. Therefore, we concluded that both recombinant proteins are immunogenic and have potential to be vaccine candidates against vivax malaria.Financing: FAPESP; PIBIC-CNPq; CAPES


FCF079-2018HYDROQUINONE INFLUENCE ON THE HUMORAL IMMUNE RESPONSE INDUCED BY VACCINATION WITH DENGUE VIRUS ENVELOPE PROTEIN

ANDRÉ VINICIUS NUNES (M)*, ISABELA FEITOSA (IC)*, CINTIA HELUANY (D)*, SANDRA FARSKY*, LUÍS FERREIRA**, EDUARDO SILVEIRA*

*Department of Clinical and Toxicological Analyses, School of Pharmaceutical Sciences, University of São Paulo; **Institute of Biomedical Sciences, University of São Paulo

Introduction and Objectives: Cigarette smoke has more than 8000 substances identified. Among them, hydroquinone (HQ) has standed out as a product of benzene biotransformation and immunosuppressive effects. Thus, would HQ-exposed individuals respond lesser to vaccines? Material and Methods: Six to 8 week-old C57BL/6 mice were daily exposed to 2500 ppm of HQ for a total of 8 weeks (1 hour/day). Animals were immunized with 10µg of recombinant protein coding for the Envelope domain III from Dengue virus serotype 2 (EDIII) adjuvanted with Alum at weeks 4, 6 and 8. Two weeks after each immunization, were evaluated for IgG titration by ELISA. At different time points after the last immunization, animals had blood draws collected and spleens and thymus removed for immunological analyses. Furthermore, a systematic gene expression analysis was conducted with public transcriptome obtained from blood samples of humans exposed to benzene or healthy counterparts.Results and Conclusions: We observed a trend towards lower titers of EDIII-specific IgG and IgG1 in HQ-exposed mice (p>0,05). Histological analyses revealed reduced area and number of splenic follicles and a significant reduction of the thymic medullary area in HQ-exposed mice in comparison to controls (p<0,05). Gene expression data pointed out that benzene-exposed people had downregulation for 8 targets related to humoral response in comparison to controls. Although the HQ-exposure presented little influence over antibody titers, it tends to alter the structure of lymphoid organs, which could be related to some compensatory mechanism in the generation of antibodies specific to EDIII. Financing: CAPES

FCF080-2018HYPERGLYCEMIA ALTERS THE BEHAVIOR OF MACROPHAGES UNDER STIMULATION BY LPS

THAIS SOPRANI AYALA (D)*, FERNANDO HENRIQUE GALVÃO TESSARO (D)*, GRASIELLE PEREIRA JANUZZI (PD)*, LEONARDO MENDES BELLA (D)*, KAREN SPADARI FERREIRA**, JOILSON DE OLIVEIRA MARTINS*

*Departamento de Análises Clínicas e Toxicológicas, FCF-USP; ** Departamento de Ciências Farmacêuticas-UNIFESP

Introduction and Objectives: In a high glucose environment, macrophages responses can be limited upon stimulation and this fact can directly contribute to infections in diabetic individuals. We examined how hyperglycemia modulates bone marrow-derived macrophages (BMDM) stimulated by lipopolysaccharide (LPS).Material and Methods: BMDM from non-diabetic and diabetic (alloxan 60 mg/kg, i.v) male C57BL/6 mice (CEUA/FCF/USP-488) were stimulated with LPS (100 ng/mL) in vitro and exposed to normal (5.5 mM) (NG) and high glucose (HG) media. TLR4 expression was evaluated by flow cytometry; phagocytosis was verified by opsonized sheep red blood cells; intracellular signalling pathways were checked by Western blot; H2O2 release was determined by amplex® red assay kit and nitric oxide (NO) measurement with GRIESS reaction.Results and Conclusions: Diabetic-BMDM stimulated with LPS presented reduced TLR4 expression on cell surface, less phagocytosis capacity, reduced levels of NO and phosphorylated more in p46 SAPK/JNK and ERK 42 MAPK compared to BMDM from normoglycemic mice. BMDM from non-diabetic mice were cultured in HG condition and stimulated with LPS presented reduced TLR4 expression on cell surface, reduced levels of NO and H2O2and an increased phosphorylation at PI3K p85 subunit. Conversely, diabetic BMDM cultured in HG presented reduced phosphorylation at PKC-d and p46 SAPK/JNK, but enhanced phosphorylation of p46 subunit at SAPK/JNK after LPS stimulation. High glucose seems to modify macrophages behaviour, affecting in different aspects in both diabetic and healthy BMDM under the same LPS stimulus. Financing: FAPESP, CAPES e CNPq


FCF081-2018SYNTHESIS OF INHIBITORS OF SIRTUIN 2 FOR CHAGAS DISEASE

EVELIN FORNARI PEREIRA (D), GUSTAVO HENRIQUE GOULART TROSSINI

Faculty of Pharmaceutical Sciences – University of São Paulo, São Paulo, Brazil

Introduction and Objectives: Neglected diseases are caused by infectious and parasitic agents. These diseases are prevalent in tropical and poor areas and they present great medical relevance. Chagas disease is endemic in Latin America and has spread to other continents by migratory movements. The currently strategy in drug discovery involves the choice and selection of specific biochemistry target. In case, sirtuin 2 (silent information regulator 2) were shown to be essential for the in vitro growth of T. cruzi.This work presents synthesis of analogues of sirtinol, designed by LBDD (ligand based drug design) to inhibit the enzyme Sir2 for Trypanossoma cruzi as antichagasic candidates. Material and Methods: General procedure: amine (1 mmol), aromatic aldehyde (1 mmol) and isatoic anhydride (1 mmol) in ethanol (5 mL). Microwave conditions: 40 minutes in 80 ºC. The products were purified by recrystallization. The basic route for the synthesis of analogs of sirtinol is a simple and innovative method for a multicomponent Schiff base reaction. This procedure was designed, planned and implemented eliminating the use or generation of substances harmful to health and the environment. Results and Conclusions: The described work was elaborated with the purpose of synthesizing 16 analogues of sirtinol, varying the amines and named sequentially by EFSi. The best time was reached to carry out the reactions and presented moderate to good yield. This is an efficient way to obtain products with better yield focusing on Green Chemistry. Yields: EFSi1 - 64%, EFSi2 - 71%, EFSi3 - 48%, EFSi4 - 55%, EFSi5 - 50%, EFSi6 - 83%, EFSi7 - 47%, EFSi9 - 66%, EFSi10 - 53%, EFSi11 - 52%, EFSi12 - 74%, EFSi15 - 54%, EFSi16 - 49%, EFSi17 - 66%, EFSi18 - 52%, EFSi19 - 49%. Financing: CNPq; FAPESP

FCF082-2018B CELL SIGNALLING ON ANTIBODY-SECRETING CELL DIFFERENTIATION IN VITRO AND IN DENGUE PATIENTS

VIVIAN BONEZI (D)*; DIÓGENES LIMA (D)*; MATHEUS BURGER (M)*; LENNON PEREIRA (D)** LUIS CARLOS FERREIRA**; MAURO CORTEZ***; HELDER NAKAYA*; EDUARDO SILVEiRA*

*Department of Clinical and Toxicological Analyses, FCF/USP; **Department of Microbiology or *** Parasitology, ICB/USP

Introduction and Objectives: Dengue is a mosquito-borne viral disease caused by four Dengue virus (DENV) serotypes. The disease affects annually about 100 million people worldwide, ranging from asymptomatic to severe forms. Dengue patients present an exacerbated blood antibody-secreting cell (ASC) response promptly after the symptoms onset. That response represents more than 50% of all circulating blood B cells, which is greater than found in ASC-leukemia patients. Furthermore, low blood calcium levels have been reported in DENV infections. Thus, our goal is to elucidate how DENV infection elicits this massive ASC response and whether calcium influences it. Material and Methods: Public transcriptome data derived from Dengue patients and healthy individuals were analyzed for the identification of genes expressed during ASC differentiation. Those targets have been quantified in protein extracts of healthy blood CD19+ B cells differentiated into ASCs with DENV-infected or uninfected CD14+ monocytes (Western blotting). The ASC enumeration and antibody titer quantification have been performed through ELISPOT, FACS and ELISA. To estimate the calcium importance in this process, BAPTA (calcium chelator), FK506 or cyclosporin A (calcineurin inhibitors) have been added to cultures. Results and Conclusions: Transcriptome data presented targets with altered expression and strong interactions with ASC pathways in both culture conditions. Those findings seem to be consistent with ASC differentiation with mitogens. The elucidation of which B cell signalling pathways are activated during DENV infection will shed light on a better understanding of that ASC response.Financing: CNPq ; CAPES


FCF083-2018NEUROCHEMICAL ANALYSIS OF COCAINE-INDUCED COUNTER-SENSITIZATION USING AYAHUASCA AS A THERAPEUTIC AGENT

VÍTOR BRUNO (M)1, VINÍCIUS NEUGEBAUER (IC)1, JÉSSICA KOIAMA (IC)1, LARISSA TORRES PACHECO2, RAPHAEL GARCIA3, ROSANA CAMARINI4, TANIA MARCOURAKIS1

1Department of Clinical and Toxicological Analyses, FCF/USP; 2Department of Food and Medicine; School of Pharmaceutical Sciences; Unifal; 3Department of Pharmaceutical Sciences, School of Pharmaceutical Sciences; Unifesp; 4Department of Pharmacology, ICB/USP

Introduction and Objectives: This study aimed to evaluate the effect of ayahuasca (AYA) treatment, after a cocaine (COC) sensitization period, on D1 and D2 dopaminergic receptors in the striatum and hippocampus.Material and Methods: Six-week-old C57Bl/6 mice were habituated in the open field (OF) for 3 consecutive days after administration of saline 0,9% i.p. Next day, the behavioral sensitization (BS) induced by COC protocol was started, in which they received, for 5 alternate days, COC 10mg/kg or saline 0,9% i.p. and were directly submitted to an evaluation of the locomotor activity in the OF. After BS protocol, the period of withdrawal or treatment with AYA began, in which they received water or AYA 3 mg of DMT/kg v.o. for 8 consecutive days. After 24h, they received a saline challenge and the next day, a COC challenge was conducted. After evaluation in OF, the animals were euthanized and had their striatum and hippocampus dissected. The structures were further prepared for analysis of D1 and D2 receptors by immunoblotting.Results and Conclusions: A significant decrease of D1 in the striatum in SSC-group was observed when compared to SAC-groups. AYA was able to significantly reduce D1 receptors after COC challenge in the striatum. This result shows that AYA is able to modulate DS in mice exposed to the BS protocol induced by COC. Financing: FAPESP, SENAD, CNPq

FCF084-2018DEVELOPMENT OF OLEOGELS BASED ON HIGH OLEIC SUNFLOWER OIL STRUCTURED BY SORBITAN MONOSTEARATE AND CANDELILLA WAX

EMANUELE DE SOUZA VIEIRA (IC); MARSIA DOLORES SERRANO SANSÓN (M); LUIZ ANTONIO GIOIELLI; JULIANA NEVES RODRIGUES RACT


Introduction and Objectives: The influence of the concentration of structuring agents (SA) sorbitan monostearate (SMS) and candelilla wax (CW) in the development of oleogels (OG) and the interaction between them was investigated. Material and Methods: The OG were prepared by structuring high oleic sunflower oil (HOSO) by CW:SMS blends in different proportions, 100:0, 80:20, 60:40, 40:60, 20:80 and 0:100, in concentrations 2, 4 and 8%. HOSO fatty acid composition was determined by gas chromatography, which allowed the calculation of its iodine and saponification values. The OG were characterized by visual analysis of the samples at temperatures between 5 and 40ºC, which were classified as liquid, viscous liquid or gel. The consistency of the OG was determined by a texture analyzer and the oil binding capacity (OBC) was based on the measurement of oil loss after centrifugation. Results and Conclusions: CW alone showed a good potential as a SA as opposed to SMS, taking into consideration that a gelled system was observed with 100:0 at 2% up to 30°C, whereas gelation only occurred at 5 and 10 °C with 8% of isolated SMS. The OBC showed that OG were capable to hold from 70 to 100% of oil. At concentrations 4 and 8%, consistency and phase diagram results revealed a positive CW/SMS interaction in the sample 60:40, which indicated a synergism in oil gelation when CW and SMS are used together in the proportion 60:40. This interaction would allow a reduction of total SA concentrations used for oleogel development, thus reducing costs and improving the sensory aspect by avoiding waxy taste. Financing: PUB-USP; CAPES; CNPq.


FCF085-2018PRODUCTION AND PARTIAL PURIFICATION OF BACTERIOCIN-LIKE INHIBITORY SUBSTANCES (BLIS) WITH ANTICARIOGENIC PROPERTIES

TAÍS MAYUMI KUNIYOSHI (PD), VITÓRIA GONÇALVES DOS SANTOS (IC), SABRINA DA SILVA SABO (PD), RICARDO PINHEIRO DE SOUZA OLIVEIRA

Department of Biochemical and Pharmaceutical Technology/School of Pharmaceutical Sciences

Introduction and Objectives: Dental caries is a disease that leads to enamel demineralization by acidogenic bacteria such as Streptococcus mutans. Recently, probiotic lactic acid bacteria (LAB) have been used to prevent this disorder through the production of antimicrobial biomolecules, such as BLIS, which can act against the aforementioned bacteria. The aim of this work was the production and partial BLIS purification produced by the LAB Pediococcus pentosaceus ET34, which was evaluated regarding its antimicrobial potential. Material and Methods: To achieve this, ET34 cells were cultivated in MRS medium and their cell-free supernatant (CFS) were evaluated against different Gram-positive and Gram-negative bacteria. The BLIS present in the P. pentosaceus ET34 CFS were partially purified with different concentrations (10-60%) of ammonium sulfate (AS). Finally, the CFS purity was evaluated on tricine SDS-PAGE gels. Results and Conclusions: The CFS of P. pentosaceus ET34 showed bactericidal effect against different Gram-positive bacteria, including Streptococcus mutans mutans UA159. The BLIS present in the CFS samples were partially purified with 20% of AS and analysed on tricine SDS-PAGE gel and it antimicrobial activity was quantified, resulting in 12,800 AU/mL. In conclusion, P. pentosaceus ET34 CFS showed antimicrobial activity against different Gram-positive pathogenic bacteria including the cariogenic bacterium S. mutans mutans UA159. Furthermore, the BLIS partial purification was successfully obtained from this CFS through salting out (20% of AS) approach. Financing: FAPESP; CNPq

FCF086-2018BIODIGESTED VINASSES FOR THE GROWTH OF PHOTOSYNTHETIC MICROORGANISMS

LAURIS DEL CARMEN MEJIA DA SILVA (PD), CÉLIA LEITE SANT’ANNA*, JOÃO CARLOS MONTEIRO DE CARVALHO

Department of Biochemical and Pharmaceutical Technology, FCF/USP *Institute of Botany, Nucleus of Phycology, SP

Introduction and Objectives: Vinasse is a waste by-product resulting from the ethanol production. It is a dark brown liquid with strong odor and high acidity (pH 3.5-5), high COD, BOD5, K, N and P. Studies have shown that photosynthetic micro-organisms have a high capacity for inorganic nutrient absorption and their biomass can be used with biofuels, biofertilizers or animal feed. The objective of this study was to evaluate the growth of Monoraphidium contortum and Synechocystis salina with nutrients such as phosphorus, nitrogen, mineral salts derived from the anaerobic treatment of vinasse.Material and Methods: M. contortum and S. salina cultures were studied in Erlenmeyer flasks by batch process, using biodigested vinasse in nature and diluted 5 and 10-fold. The kinetic parameters were compared in the different experimental conditions adopted for the growth of the photosynthesizing microorganisms in the vinasse treated as in nature and diluted.Results and Conclusions: Biodigested vinasse diluted 5-fold favored the growth of M. contortum and S. salina with maximum dry biomass concentrations of 702 and 520 mg.L-1 respectively, and biodigested vinasse diluted 10-fold was favorable for S. salina with maximum dry biomass concentrations of 512 mg.L-1. Microscopic observations revealed that in nature vinasse was not favorable for the growth of the strains, pure vinasse may be toxic, due to presence of residual compounds of biodigestion or presence of minerals at higher concentrations. The use of biodigested vinasse may be feasible for biomass production of photosynthetic microorganisms, reducing the cost of production by the use of nutrients such as phosphorus and nitrogen. Financing: CAPES


FCF087-2018PHYSICAL-CHEMICAL QUALITY CONTROL STUDY OF THE PURIFIED WATER USED IN CONFAR ANALYTICAL PROCEDURES

ILDEMARA GRAÇAS DE SOUZA (IC), GABRIELA CORRÊA CARVALHO (PG), MARINA DE SOUZA BRAGA (PD), ROGERIO TAKAO OKAMOTO, FELIPE REBELLO LOURENÇO, TEREZINHA DE JESUS ANDREOLI PINTO

Department of Pharmacy - Faculty of Pharmaceutical Sciences - University of São Paulo. CONFAR Laboratory - USP

Introduction and Objectives: The objective of this study was to monitoring the physical-chemical quality control of the purified water used in the Laboratory of control of Medicines, Cosmetics, Domisanitary Products, Related Products and their Respective Raw Materials (CONFAR) in order to reduce the periodicity of pH and conductivity analyzes. These tests aim to ensure the water purity, as values out of specification which characterize contamination interfere in the result of further analyzes.Material and Methods: So, conductivity and pH evaluation were daily performed in six water samples: two collected from storage kegs and four from reverse osmoses equipment. Five months results were considered (November/2017 to March/2018) and submitted to a statistical analysis. Media, standard deviation and capability index (Cpk) of pH and conductivity values were calculated. Results and Conclusions: All of them were within specification and the CpK value is superior to 1.3, which indicates that the risk of producing out of specification water is very low. Therefore, it is concluded that the periodicity of water collection for pH and conductivity analyzes may be longer, since the results show that the water production system of CONFAR laboratory is robust and does not present a statistically significant variation. Financing:

FCF088-2018UNDERSTANDING SKIN EPIDERMIS ALTERATION TO AIR PARTICULATE MATTER 10 (PM10) IN A RECONSTRUCTED HUMAN EPIDERMIS (RHE) MODEL

DANIELE SEO HIEDA* (PD); BARBARA VAZ DE MELLO* (IC); SILVIA ROMANO DE ASSIS*; JOANNA WU**; LAURENCE DU-THUMM**; SILVYA STUCHI MARIA-ENGLER*; SILVIA BERLANGA DE MORAES BARROS*

*Faculty of Pharmaceutical Sciences, USP, Brazil **Colgate-Palmolive, USA

Introduction and Objectives: This study investigated the alterations induced by the exposure of a RHE to air PM10 in proteins of differentiation and water transport of the skin. Material and Methods: The RHE was exposed to 2.2, 8.9 and 17.9 µg/cm2 of a standard Urban Particulate Matter for 24 and 48h. Cell viability, epidermis barrier function, IL-1a and morphology (HE) and some protein markers by immunohistochemistry (IHQ) were evaluated. Results and Conclusions: Cell viability decreased after 48h of exposure to the two highest doses. Barrier function also decrease after 24 and 48h of exposure to 17.9 µg/cm2 of PM10. IL-1a release increased in all treatments with the higher response after 48h of treatment. Morphological alterations included cytoplasm vacuolization and loss of stratum corneum after treatment with the higher dose. Cytokeratin 10 indicates the good stratification of the RHE however reduced expression was observed after the treatment with the two highest doses, as with filaggrin, involucrin and loricrin. Aquaporin 3 was increased in RHE exposed to PM10 in comparison to control both by IHQ and Western Blot. This last observation can lead to increased water transport to the stratum corneum contributing to water loss and dry skin as observed in atopic eczema. In conclusion, the exposure of a RHE to PM10 induces dose dependent alterations in the epidermis with loss of proteins at granular and cornified layers and increase in aquaporin 3 suggesting alterations of barrier function and water transport across the epidermis with possible epidermis dehydration. Financing: FAPESP; Sponsored by Colgate Palmolive USA


FCF089-2018PRELIMINARY STUDIES ON BIOPROSPECTING PROBIOTIC LACTIC ACID BACTERIA PRODUCING VITAMIN B COMPLEX AND BACTERIOCINS FROM CHICKEN CECUM

SABRINA DA SILVA SABO (PD)*, TEREZINHA KNÖBL**, RICARDO PINHEIRO DE SOUZA OLIVEIRA*

*Department of Biochemical and Pharmaceutical Technology, FCF/USP **Veterinary Pathology Department, FMVZ/USP

Introduction and Objectives: The use of probiotic in the prevention and treatment of infections in animals intended for human consumption has been considered an efficient alternative to the use of antibiotics. It is known that the beneficial effects generated by probiotics are host-specific and, frequently, interest compounds are synthesised in greater quantity by a given bacterial strain. The objective of this study was bioprospecting probiotic lactic acid bacteria (LAB) present in the intestinal microbiota of chicken with high capacity to produce bacteriocins and vitamins B complex (i.e., riboflavin and folate).Material and Methods: 10 g of cecum were diluted in 90 mL sterile saline and plated onto de Man, Rogosa, and Sharpe and Bifidus Selective Medium agar plates for selecting LAB. In order to investigate riboflavin and folate producers, the isolates were grown in Riboflavin Assay Medium and Folic Acid Casey Medium, which are free from riboflavin and folate, respectively. For bacteriocin production, cell-free supernatants were collected from isolates culture broth and tested against different bioindicator strains.Results and Conclusions: From 314 isolates, 35 of them produced antimicrobial activity against tested bioindicator strains, including Salmonella enterica, one of the most important poultry pathogens. 8 of these 35 isolates were investigated regarding the production of the aforementioned vitamins. From these 8, 5 of them produced folate and 4 isolates produced riboflavin. Interestingly, 3 of these 8 isolates produced both vitamins and bacteriocin simultaneously, which represents potential strains to be employed as probiotic bacteria in feed for poultry, as future studies. Financing: FAPESP

FCF090-2018CHEMICAL RISK: CRIAÇÃO DE UM JOGO DIDÁTICO PARA O ENSINO DE BIOSSEGURANÇA

LUIZ FERNANDO DE SOUZA2 (IC); DANIEL MARTINS DA SILVA OLIVEIRA2 (IC); GLEDSON KLEITON DA SILVA2

(IC); WILKER ALVES DE MOURA2 (IC); ÁLVARO GABRIELE RODRIGUES2 (IC); CRISTINA NORTHFLEET DE ALBUQUERQUE1

1Departamento de Tecnologia Bioquímico-Farmacêutica da FCF/ USP. 2FATEC Carapicuíba, Curso dde Design de Games

Introduction and Objectives: Chemical Risk is a didactic project that aims to demonstrate the use of games in education. Its objective is to establish the main concepts of biosafety, this project proposal deals with the familiarization of the student with such concepts. Refers to a reflection on the traditional model of education and proposes a new concept allied to technological evolution suggesting new ways of transmitting knowledge. It provides players with a learning practice about biosafety concepts, as it uses the simulation of a chemical accident in a laboratory environment, allowing the student to apply and consolidate the concepts seen in the classroom. As a bibliographic basis, this project sought references in academic books, online portals and articles on the use of games in education and learning through computer games. Material and Methods: Softwares: Windows 7 (Professional, Ultimate), Microsoft Office 2010 Enterprise, Blender3d, Unity3d, Autodesk Maya 3d, SketchUp 8 e SketchUp Pro 8; Hardwares: Notebook LG A410 – Core I3 2.67GHz – 4GB RAM – 750GB – NVIDIA GeForce 310M e Notebook Dell Latitude 14 – Core I5 2520M 2.5GHz – 4GB RAM – 300GB. Results and Conclusions: After the classroom tests, the general observation was that traditional lessons are needed to understand the basics, but the use of games such as exercises and content-fixing tools is extremely positive and very relevant. Financing: Sem auxílio financeiro.


FCF091-2018BLIs PRODUCTION BY PROBIOTIC CULTURES: ACTIVITY AGAINST Listeria innocua

ANNA CAROLINA MEIRELES PIAZENTIN (D); RICARDO PINHEIRO DE SOUZA OLIVEIRA


Introduction and Objectives: Bacteriocins or bacteriocin-like inhibitory substances (BLIs) are peptides or proteins that exert antimicrobial activities against other bacterial species. Among the main LAB (lactic acid bacteria) producing BLIs, are Lactobacillus spp. these bacteria are commonly used in food fermentation, but are already known for their antimicrobial effects. The present work evaluates the growth and production of BLIs of two strains of Lactobacillus spp. and its activity against Listeria innocua.Material and Methods: The strains of L. salivarius and L. reuteri were grown in MRS medium pH 6.0 on agitation at 100 rpm 35 °C, after 24h of growth they were centrifuged at 3,857 RCF for 15 minutes and their supernatant was filtered at 0.45 µm, after that its pH was modified to 6.0-6.5 and placed in a bath at 80 °C for 10 minutes. To verify the antimicrobial activity 20 µL of it step of the supernatant treatment were added to a Petri plate containing Listeria innocua, the plate was maintained at 37 °C for 18 hours to verify the formation of inhibition halos. Results and Conclusions: After 24 hours of cultivation, a cell mass production of 3.23 g/L and 2.22 g/L was observed for L. salivarius and L. reuteri respectively, however the antimicrobial activity presented was higher in L. reuteri, the same showed inhibition of Listeria growth in the three conditions tested, with halos of 24 mm, otherwise L. salivarius showed halos of 13 mm, and did not present activity in the samples that suffered the bath. We can conclude that both strains used have anti-listeric activity, and that possibly the BLIs produced by L. salivarius is thermosensitive.Financing: CNPq

FCF092-2018COMPARISON OF ATTRIBUTES OF CHOCOLATES PRODUCED WITH BRAZILIAN AND PERUVIAN COCOA LIQUORS

VIVIAN SIOU SAN CHOW (IC); INGRYD CAROLINNE COSTA CAMPOS (D); SUZANA CAETANO DA SILVA LANNES

Pharmaceutical-Biochemical Technology Department, Pharmaceutical Sciences School, University of São Paulo (USP), São Paulo, São Paulo, Brazil

Introduction and Objectives: Dark chocolates are promising sources of bioactive and may offer beneficial cardiovascular effects. Material and Methods: To compare alkalinized Brazilian and Peruvian liquors and derived chocolates, relating to physical and chemical characteristics, dark chocolate was produced in a ball mill Universal Equipment (Mazzetti, Italy). Results and Conclusions: The composition of the Brazilian liquor used was similar to Peruvian’s, except for humidity (1.42 ± 0.04% for Brazilian and 3.96 ± 0.48% for Peruvian), lipids (52.12 ± 0.35% and 50.48 ± 0.94%) and carbohydrates (30.04% and 26.84%). The physicochemical attributes of these liquors differ at: water activity (0.291 ± 0.008 for Brazilian and 0.512 ± 0.007 for Peruvian) and maximum particle size (0.042 ± 0.005 mm and 0.432 ± 0.148 mm). Dark chocolate made with Brazilian liquor showed similar humidity value compared to dark chocolate made with Peruvian liquor, but they differ at lipids (32.79 ± 2.06% for Brazilian and 39.35 ± 2.06% for Peruvian) and proteins (16.32 ± 1.89% and 10.21 ± 1.14%), totalizing different energetic values. In relation to reducing compounds, it is estimated that Peruvian liquor (552.44 mg Trolox / g liquor) has a higher potential than the Brazilian’s (278.41 mg Trolox / g liquor). The same should be reflected, therefore, in their respective chocolates. Peruvian liquor has a higher composition of antioxidants that are more capable of reducing iron ions (6714.81 µM ferrous sulphate / g liquor) than other free radicals while Brazilian’s reduced 4083.70 µM ferrous sulphate / g liquor. Still, both liquors proved suitability for industrial production by producing in a ball mill equipment.Financing: Unified Scholarship Program (PUB-USP)


FCF093-2018DEVELOPMENT AND VALIDATION OF KEY PERFORMANCE INDICATORS FOR MEDICATION MANAGEMENT SERVICES PROVIDED FOR OUTPATIENTS

TÁCIO DE MENDONÇA LIMA (D), PATRICIA MELO AGUIAR, SÍLVIA STORPIRTIS

Department of Pharmacy, Faculty of Pharmaceutical Sciences, University of São Paulo, São Paulo, Brazil

Introduction and Objectives: One of the elements of quality improvement of medication management services is measuring the quality of care and key performance indicators (KPIs) can be used in this assessment. There has been no publishing yet about a KPI instrument developed for ambulatory setting. The study is aimed to develop and validate KPI instrument for medication management services provided for outpatients. Material and Methods: A working group established 7 possible KPIs for assessment of the expert panel. For content validity, 16 experts were invited to participate in an internet based 2-round Delphi approach. Expert panel rated 7 possible KPIs using 7 attributes on a 5-point Likert scale for consensus. In order to construct validity and reliability, an internet questionnaire was developed for pharmacists that work in primary care to understand the staff views. In addition, a comparison between expert and pharmacist views about the indicators relevance was performed. Results and Conclusions: Eleven (68.8%) experts participated in the Delphi round 1 and nine (81.8%) experts completed the 2 Delphi rounds. A new KPI was develop after expert panel assessment in the first round. Overall, content and construct validity were reached for 6 KPIs: pharmaceutical consultation provided (I1), pharmacist interventions accepted by the prescriber (I2), drug therapy problems resolved (I3), patient clinical status (I4), patient satisfaction (I5), and patient quality of life (I6). It is expected that these KPIs will improve the quality of medication management services.Financing: CAPES

FCF094-2018OIL-IN-WATER VEGETABLE NANOEMULSION USING D-PHASE EMULSIFICATION AS A UNIQUE LOW-ENERGY PROCESS

MEGUMI NISHITANI YUKUYAMA (M), NÁDIA ARACI BOU-CHACRA


Introduction and Objectives: D-phase emulsification (DPE) is a less known and more recent alternative nanoemulsion (NE) preparation process when compared to conventional low energy processes, such as Phase Inversion Temperature (PIT) and Phase Inversion Composition (PIC) methods. Employing DPE with Box-Behnken statistical approach, a stable 20 – 30 nm oil-in-water (O/W) NE containing 40.0 % olive oil was successfully obtained using only 2.0 % of oleth-20 as a surfactant (all w/w). The aim of this study was to optimize the DPE method by a statistical approach and to evaluate the influence of the polyol in this specific low-energy process.Material and Methods: The material comprises oleth-20 (Croda), olive oil and glycerin (Sigma Aldrich) and ultrapure water. The preparation by DPE process was based on Endoo and Sagitani (1991) work, by dropwise addition of components under stirring with a magnetic bar, at specific speed and temperature. The study was carried out by using Box-Behnken experiment to evaluate the influence of process and composition variables on the NE mean particle size (MPS).Results and Conclusions: The polyol (glycerin) was confirmed to be essential as a variable influencing the MPS of NE in this specific process. Under an established condition such as temperature, concentrations of initial water and surfactant, a very narrow concentration range of glycerin (2.0 – 3.0 % w/w) has been shown to produce an O/W NE below 100 nm. An isotropic phase was created rather than a liquid crystal or a conventional microemulsion during the transition phase. A stable NE with 27.74 ± 0.66 nm with 40.0 % (w/w) olive oil was successfully obtained. In addition, the DPE method did not require a strict adjustment of the hydrophilic-lipophilic-balance during the preparation of nanoscaled emulsion. Financing: CAPES


FCF095-2018CHANGE IN SERUM POTASSIUM IS CORRELATED WITH SPECIFIC MICROBIOME GENERA IN CHRONIC KIDNEY DISEASE PATIENTS UNDERGOING UNRIPE BANANA FLOUR CONSUMPTION WITHOUT FURTHER DIETARY CHANGES

LOUISE RIZZUTO GOMES (IC), FABIANA ANDREA HOFFMANN-SARDA (PD), IARA GUMBREVICIUS (M), ELIANA BISTRICHE GIUNTINI (PD), ELIZABETE WENZEL DE MENEZES, CHRISTIAN HOFFMANN


Introduction and Objectives: Studies have shown that resistant starch ingestion slows down damage to kidney tubules as well as intestinal interstitial fibrosis, in association with changes in the gut microbiome, in animal model. The aims of this study were to analyse change in dietary patterns, intestinal function, microbiome composition and blood biochemical parameters before and after an intervention with unripe banana flour (UBF) in CKD constipated patients undergoing hemodialysis.Material and Methods: A single-blind, placebo controlled study was designed where patients consumed UBF 3 times a week for 6 weeks. Nutrient intake was evaluated using 24h recall questionnaires and the Brazilian Food Composition table (TBCA). Microbiome profiling was determined using 16S rDNA sequencing in the Illumina platform. All measurements were taken prior to the intervention and at the completion of the study. R environment was used for statistical analysis.Results and Conclusions: There were no changes in the dietary intake pattern, or nutrients and minerals intake at the end of the intervention, for both UBF and placebo groups. Serum potassium was decreased (p=0.006, linear mixed effects model) and constipation decreased (p=0.01, Wilcoxon rank sum test) in the patients receiving UBF. A negative correlation was observed between serum potassium and bacterial genus Epulopiscium (p=0.006, Spearman correlation). UBF could be indicated as a dietary adjuvant to improve clinical parameters in CKD patients. Financing: PUB-USP, FoRC (FAPESP), Capes

FCF096-2018BEHAVIORAL EVALUATION OF AYAHUASCA IN A CONTEXT OF COCAINE-INDUCED SENSITIZATION IN C57Bl/6 MICE

VINICIUS RODRIGUES NEUGEBAUER(IC)¹, VITOR BRUNO(M)¹, JESSICA KOIAMA(IC)¹, ROSANA CAMARINI², TANIA MARCOURAKIS¹

1Department of Clinical and Toxicological Analyses, FCF/USP;2Department of Pharmacology, ICB/USP

Introduction and Objectives: Cocaine is a substance with psychostimulant properties that may lead to addiction. The objective is evaluate the possible therapeutic potential of ayahuasca in the model of behavioral sensitization (BS) to cocaine (COC). Material and Methods: (CEUA/FCF 33.2016-P518) C57Bl/6 were habituated in the open field (OF) for 3 consecutive days, after receiving saline (SAL) i.p. On the 4th day, the BS protocol was initiated, in which they received a dose of COC 10 mg/kg or SAL i.p., and were immediately placed in the OF for 5 alternate days, 24h later, the treatment with AYA at the dose of 3.0 mg DMT/kg or SAL was started, for 8 consecutive days. Locomotor evaluation at OF was performed after 30 min of AYA administration, 24h after the end of the treatment period, animals were challenged with SAL i.p. and, 24h later, were challenged with COC 10 mg/kg, and posed to the OF. The total locomotor activity was evaluated, during all experimental days for 30 min. Results and Conclusions: During the protocol of cocaine-induced sensitization, the progressive increase in the locomotor activity of the mice treated with COC was observed. On the 2nd COC administration day, the acquisition of BS was perceived, since the locomotor activity of the animals was significantly higher on this day, when compared to the previous day (*p<0.05; F(2.22) =0.3138). The administration of AYA at the dose of 3.0 mg DMT/kg did not significantly change the locomotor activity of the mice. When performing a challenge with COC 10 mg/kg, cocaine- sensitized mice treated with AYA presented similar locomotor activity to the control group. Treatment with AYA was not able to reverse the BS process triggered by the acquisition of COC sensitization. Financing: SENAD;FAPESP.


FCF097-2018STUDY OF THE PRODUCTION CONDITIONS OF BACTERIAL L-ASPARAGINASE IN Pichia pastoris

DANIELE CHUN HO (IC), GISELE MONTEIRO


Introduction and Objectives: The strain of Pichia pastoris genetically modified (Glicoswitch ® SuperMan5 his- (Biogrammatics Inc.)) produces two proteoforms of asparaginase, which differ by the glycosylation pattern. The objective of this work is to verify if the oxygen limitation favors the production of the less glycosylated enzyme, which has more activity. Material and Methods: P. pastoris was cultured in BMGY (Buffered complex medium containing glycerol) medium, in bioreactor, without oxygen control and in 30% of aeration, for 4 days. It was also cultured in 250 mL erlenmeyer flask, to study the influence of the presence of baffle and, in 2L erlenmeyer flask, the value of the agitation speed (50 rpm, 150 rpm, 250 rpm) during the induction with 1,5% methanol, in shakers incubator, at 30°C. The activity assay was made by the hydroxylamine method and the purification, by ion exchange chromatography. The determination of the glycosylation level was done by gel electrophoresis. Results and Conclusions: It was verified that in non- baffled Erlenmeyer flasks the expression of the less glycosylated enzyme was superior as well as the activity concentration. However, the cultivation forcibly limited in 30% of aeration exhibit less activity and cellular growth than that with no oxygen control. Furthermore, there was not expression of asparaginase in flask cultivation with agitation of 50 rpm during the induction. Keeping the agitation in 150 rpm results in the production of the proteoform under study, but the activity value is lower than that with agitation maintained in 250 rpm. These results indicate that the strong limitation of oxygen is disadvantageous for the expression of asparaginase. Financing: FAPESP

FCF098-2018IDENTIFICATION OF PROTECTIVE B CELL ANTIGENS FROM ASEXUAL BLOOD FORMS OF PLASMODIUM PARASITES THROUGH MONOCLONAL ANTIBODIES

MARIANA DOMINGUEZ1 (PD), IRENE SOARES1, RICARDO GAZZINELLI3, DANIEL BARGIERI2, EDUARDO SILVEIRA1

1Department of Clinical and Toxicological Analyses, School of Pharmaceutical Sciences, University of São Paulo. 2Department of Parasitology, Institute of Biomedical Sciences, University of São Paulo. 3Division of Infectious Disease and Immunology, University of Massachusetts Medical School, USA

Introduction and Objectives: Although malaria disease can be fatal, individuals from endemic areas end up developing protective immunity years after continuous exposure to malaria parasites (Plasmodium). Antibodies specific to antigens from asexual blood forms of the parasite have a critical role in that protection. Therefore, our main aim is to identify novel protective Plasmodium antigens from asexual blood forms through monoclonal antibodies (mAbs) secreted by single antibody-secreting cells (ASCs) of immune mice. Material and Methods: To generate those mice, we challenged C57Bl/6 mice with 5 P. berghei ANKA infected red blood cells (RBCs) and whenever parasitemia reached 0.5-1% of total RBCs, they received a sub-curative chloroquine treatment. After few rounds of treatment, these mice did not bounce their parasitemia levels back, becoming immune to malaria. This allowed us to continuous with the necessary assays in order to clone their mAbs. Results and Conclusions: After mice become immune, we enumerated Plasmodium berghei-specific ASCs at different time points after re-challenge through ELISPOT. We observed that the higher frequency of IgG+ ASCs on blood and spleen occurred 10 days post re-challenge. Now, we are performing single cell sorting of these ASCs for cloning the variable region of Ig gene and mAb production. After testing these mAbs regarding specificity and functionality (neutralization capacity), the positive mAbs will have their targeted antigens solved by mass spectometry. Financing: Funded by FAPESP; CAPES.


FCF099-2018L-ASPARAGINASE SUPERACTIVITY AT LOW CONCENTRATIONS OF IONIC LIQUIDS

ELOÍSA GUEDES CAVICHON* (M), JOÃO HENRIQUE PICADO MADALENA SANTOS*,** (D), ADALBERTO PESSOA JÚNIOR*, SÓNIA PATRÍCIA MARQUES VENTURA** (PD), CARLOTA OLIVEIRA RANGEL YAGUI*

*Department of Pharmaceutical-Biochemical Technology, FCF/USP **Department of Chemistry, University of Aveiro, CICECO/UA

Introduction and Objectives: The enzymatic activity of Escherichia coli L-asparaginase (ASNase), a biopharmaceutical, has been studied in ionic liquids (ILs), alternative green solvents with potential for enzymatic stability. The influence of alkylidine chain size on the activity of the ASNase enzyme of the ILs of the family was analyzed using 1-ethyl-3-methylimidazolium ([C2mim]Cl), 1-butyl- 3-methylimidazolium chloride ([C4mim]Cl), 1-hexyl-3-methylimidazolium chloride ([C6mim]Cl), 1-methyl-3-octylimidazolium ([C8mim]Cl), chloride methylimidazolium ([C10mim]Cl) and 1-dodecyl-3-methylimidazolium chloride ([C12mim]Cl). Material and Methods: Aqueous solutions of ILs (2.5, 5, 10, 25 and 50 wt%) were tested with an ASNase concentration of 0.1 mg.mL-1; enzyme activity was measured through the L-aspartic ß-hydroxamate acid (AHA) method. Results and Conclusions: ASNase activity increased in the presence of all ILs at lower concentrations (2.5 wt%). Higher activity values were observed in the presence of ILs presenting more hydrophilic cationic moieties. Activity decreased with the increase in ionic liquid concentration and length of the alkyl chain of the cation. We attributed the decrease in ASNase activity to the IL tensioactive effect on the amino acids sequence of protein, which might have resulted in conformational changes, culminating in ASNase unfolding. In addition, very high concentrations of ILs (25 wt%) resulted in enzyme denaturation by a surfactant-like effect caused by ILs micelles formed. ILs of lower alkyl chains ([C2mim]Cl and [C4mim]Cl at concentrations up to 5 wt% resulted in the best results, with ASNase superactivity.Financing: CAPES; FAPESP; CNPq; FCT/MEC

FCF100-2018PHARMACEUTICAL SERVICES IN PRIMARY HEALTH CARE: AN EXPERIENCE IN THE CITY OF SAO PAULO

SAMARA JAMILE MENDES* (D), SILVANA NAIR LEITE**, SILVIA STORPIRTIS*

* Department of Pharmacy, FCF/USP ** College of Pharmacy, Federal University of Santa Catarina

Introduction and Objectives: In Brazil, it is necessary to expand the discussion about the potentialities and weaknesses of the Pharmaceutical Services (PS), in Primary Health Care from the perspective of pharmacists. This study aimed to analyze the perception of pharmacists who work in Primary Health Care (PHC) in the city of São Paulo, in relation to pharmaceutical services.Material and Methods: It is study realized with 116 pharmacists from the Municipal Health Department (SMS-SP) who answered an online form. The PS were addressed about the achievement, frequency, degree of current importance and expectation for an ideal scenario. The answers were analyzed in a descriptive way. The study was approved by the Research Ethics Committee of the Faculty of Pharmaceutical Sciences of Sao Paulo. Results and Conclusions: Services related to the control of the stock of medicines; guidance to patients on access to and use of medicines; control of psychotropic drugs; storage of medicines pharmacy team training; dispensing of medicines and other materials; supervision of staff and information to other professionals about medications are performed by 90% of the sample pharmacists, with daily frequency. However, in the perception of pharmacists there is also an expectation that their services will have a higher degree of importance in an ideal scenario. It was possible to analyze the PS performed in PHC in the city of São Paulo, which pointed to the need for these studies in different realities of Brazil PHC. It was also highlighted the importance of studies on the perception of pharmacists, since they are the key for the process. In addition, on-site surveys are necessary with other methods that explore PS in greater depth.Financing: Not applicable


FCF101-2018BACTERIOCIN-LIKE INHIBITORY SUBSTANCES (BLIS) PRODUCTION BY PEDIOCOCCUS PETOSACEUS ET34 USING HYDROLYZED SUGAR CANE BAGASSE AS CARBON SOURCE

TAÍS MAYUMI KUNIYOSHI (PD), VIVIANE BORGES VIERA (M), RICARDO PINHEIRO DE SOUZA OLIVEIRA

Department of Biochemical and Pharmaceutical Technology/FCF-USP

Introduction and Objectives: One of today’s great challenges from the scientific area is the search of new compounds in order to substitute antibiotics. Bacteriocins are peptides produced by a number of bacteria that harbor bacteriostatic or bactericide effect over pathogenic and deleterious microorganism present in industrial processes. In this context, the aim of this work was to study the BLIS production by Pediococcus pentosaceus ET34 using hydrolyzed sugar cane bagasse as substrate. Material and Methods: The gene that codifies pediocin from ET34 was sequenced and a global alignment revealed 100% of identity with the commercial pediocin PA-1 sequence using ClustalW software. The inhibition spectrum from cell free supernatant of ET34 was determined using different bioindicator lineages through spot-on-lawn assay. Besides, it was verified that BLIS lost its antimicrobial activity when treated with proteolytic enzimes (1mg/mL of tripsin, papain or pepsin) indicating the proteinaceous nature of this compound. Results and Conclusions: Experimental data revealed that the factors which influenced the BLIS production are: hydrolyzed sugar cane bagasse concentration, temperature and pH. On the other hand, the agitation did not affect BLIS production by ET34 cells under conditions tested. In order to maximize BLIS production, cells should be cultivated with 15% sugar cane liquor at 35 ºC and pH 7.0. In conclusion, it was obtained successfully BLIS production by P. pentosaceus ET34 using hydrolyzed sugar cane bagasse as carbon source. Financing: FAPESP; CAPES

FCF102-2018PERMEABLE POLYMERSOMES ENCAPSULATING L-ASPARAGINASE

CECILIA ZORZI BUENO (PD)*, ADALBERTO PESSOA-JR*, CARLOTA DE OLIVEIRA RANGEL-YAGUI*, GIUSEPPE BATTAGLIA**

* Department of Biochemical and Pharmaceutical Technology, University of São Paulo, São Paulo, SP, Brazil ** Department of Chemistry, University College London, London, United Kingdom

Introduction and Objectives: L-asparaginase (ASNase) is an enzyme used in the treatment of acute lymphoblastic leukaemia that catalyses the hydrolysis of L-asparagine, an essential amino acid to tumour cells. However, L-asparaginase is associated with various side effects and may be inactivated by proteases and antibodies. ASNase stability, bioavailability and biocompatibility may be improved by its encapsulation in polymeric nanovesicles (polymersomes). In this work we developed polymersomes permeable to L-asparagine, while keeping ASNase encapsulated and protected against a hostile environment. Material and Methods: Polymersomes were obtained by the film-hydration method, mixing two different copolymers: PMPC25-PDPA70 (poly((2-methacryloyloxy) ethyl phosphorylcholine)- block-poly (2-(diisopropylamino) ethyl methacrylate), which forms robust membranes, resistant to interaction with plasma proteins; and PEO16-PBO22 (poly(ethylene oxide)-block-poly(butylene oxide), which forms thin membranes, permeable to small polar molecules. ASNase was encapsulated through electroporation and non-encapsulated enzyme was removed by size-exclusion chromatography. Samples were characterized using dynamic light scattering, transmission electron microscopy and high-pressure liquid chromatography. Results and Conclusions: PMPC-PDPA polymersomes containing a small PEO-PBO bud were obtained, with average mean diameter of 112-120 nm and PdI of 0.17-0.20. ASNase was encapsulated with loading efficiencies of 740-845 molecules per vesicle. The ASNase loaded permeable polymersomes are promising for LLA treatment by protecting the enzyme against proteases and minimizing side effects. Financing: FAPESP; CNPq


FCF103-2018DEFEROXAMINE MESYLATE AS A NEW ADJUVANT FOR PRESERVING PHARMACEUTICAL AND COSMETIC FORMULATIONS

MÁRCIA LOMBARDO (D), JAQUELINE KALLEIAN ESERIAN*, FELIPE REBELLO LOURENÇO, TELMA MARY KANEKO

Department of Pharmacy, FCF/USP; *Instituto Adolfo Lutz, SP

Introduction and Objectives: Preservatives and chelating agents are adjuvants added in a wide range of products, in order to avoid the degradation of the formula and the risks to the user’s health. Still, many of the adjuvants commonly used for the preservation of medicines and cosmetics are related to hypersensitivity and negative impacts on the environment, prompting the search for new agents. In this work, we investigated the antimicrobial activity of preserving systems containing low concentrations of deferoxamine mesylate (DFO), a chelating agent used in the iron overload therapy.Material and Methods: A factorial design of experiments (DoE) was drawn up to evaluate different combinations of ethylenediamine tetra acetic acid (EDTA), DFO, bisulphite and methylparaben as a preservative formulation system. Antimicrobial activity was assessed by broth microdilution technique, against microorganisms recommended by US Pharmacopeia. DoE analysis were performed using Minitab statistical software and it was adopted a significance level of 5% (p = 0.05).Results and Conclusions: The mathematical models obtained for the growth inhibition of each microorganism showed high prediction accuracy. DFO concentrations up to 0.1% w/v revealed synergistic responses when associated with bisulphide (S. aureus and E. coli), EDTA (E. coli, P. aeruginosa and C. albicans) or methylparaben (A. brasiliensis). These results may contribute to microbiological stability of pharmaceutical and cosmetic formulations and with the improvement of the quality of products. Such finds are particularly interesting when the objective of reducing preservatives in formulations is considered.Financing:

FCF104-2018ISOLATION AND IDENTIFICATION OF LACTIC ACID BACTERIA PRODUCING BACTERIOCIN-LIKE INHIBITORY SUBSTANCES (BLIS) FROM MILK SAMPLES

VIVIANE BORGES VIERA (M), TAÍS MAYUMI KUNIYOSHI (PD), RICARDO PINHEIRO DE SOUZA OLIVEIRA

Department of Biochemical and Pharmaceutical Technology / FCF-USP

Introduction and Objectives: Lactic-acid bacteria are currently classified as part of the Gram-positive bacteria group which produce high added value bioproducts for the food industry through their fermentation process. The Bacteriocins are peptides described in the literature for their antimicrobial properties against certain microorganisms. The aim of this work was isolating LAB bacteriocing-producers from milk samples and propose a pre-purification technic. Material and Methods: Bacteria were isolated from the milk and planted directly on Man, Rogosa and Sharpe (MRS) and agar M17. The isolate that showed zones of inhibition against Listeria innocua was selected for identification of MALDI-TOFMS and sequencing of the 16S rRNA gene. The cell-free supernatant (CFS) of the isolate was treated with proteolytic enzymes (1mg/ml), moreover pre-purified by precipitation with 40% ammonium sulfate (SA) and analyzed on SDS-PAGE tricine for visualization of peptides.Results and Conclusions: Results MALDI-TOFMS revealed that the isolate belongs to Lactococcus genus. In addition, sequencing of the 16S rRNA gene showed 99% identity with Lactococcus lactis.CFS treated with proteolytic enzymes did not exhibit a zone of inhibition demonstrating its protein nature. The one precipitated with SA generated a pronounced band about 2- 5 kDa in tricine-SDS-PAGE which exhibit an effective antimicrobial activity against Listeria innocua. These results demonstrated that LAB producers of bacteriocin from the milk isolate were successfully obtained. Despite the fact that it is not possible to determine that bacteriocin corresponds to this antimicrobial substance produced by Lactococcus lactis, the compound has been shown to be a peptide. Financing: CAPES; FAPESP.


FCF105-2018PRODUCTION AND PURIFICATION OF L-ASPARAGINASE I SMALL OF SACCHAROMYCES CEREVISIAE

CAROLINA SILVA (IC); IRIS MUNHOZ COSTA (D); GISELE MONTEIRO DE SOUZA

Department of Biochemical-Pharmaceutical Technology, School of Pharmaceutical Sciences of University of São Paulo, SP, Brazil

Introduction and Objectives: L-asparaginase is an enzyme used in the treatment of acute lymphocytic leukemia (ALL). Although its high therapeutic effectiveness L-asparaginase commercially available, obtained from bacteria, have high rates of immunogenic reaction. Therefore, there is a search for alternative sources of the enzyme. Recently, L-asparaginase I (ASP1) of Saccharomyces cerevisiae has been successfully expressed in Escherichia coli. High molecular weight protein complexes with histidine tag were obtained, which after processing for tag removal, resulted in a low molar mass monomer enzyme, called small. Owing to its relatively small size (~ 42kDa), ASP1 small may have low immunogenicity. The aim of the project is to evaluate the oligomeric state and activity of ASP1 of S. cerevisiae with no tag fusion expressed in E. coli. Material and Methods: The ASP1 gene was inserted in the pET-22b vector, between restriction site BamHI and NdeI and cloned into E. coli DH5a for plasmid selection. The protein has been expressed in E. coliBL21 (DE3), induced with IPTG. The expression was confirmed by SDS-PAGE. The purification of the protein of interest was performed by gel filtration chromatography and analyzed by Native-PAGE. Results and Conclusions: The expression plasmid was successfully constructed and the enzyme of interest was super expressed in E. coli BL21 (DE3). Under the conditions tested, the protein was expressed in soluble form. The purification by gel filtration resulted in ASP1 in different oligomeric forms, including the monomeric one. The results show that monomer form can be obtained directly, without the processing steps to remove the histidine tag. Financing: FAPESP

FCF106-2018INVESTIGATION OF MICRONIZATION PROCESS ON PHYSICOCHEMICAL CHARACTERISTICS OF ACETAZOLAMIDE: DEFINITION OF DESIGN SPACE BY STATISTICAL PLANNING APPROACH

ERIKA KISHIMA (IC),KATHERINE JASMINE CURO MELO (D),VINICIUS DANILO NONATO BEZZON (PD), MICHELE GEORGES ISSA(PD),HUMBERTO GOMES FERRAZ


Introduction and Objectives: Acetazolamide (ACZ) is a drug used in the treatment of glaucoma, epilepsy, edemas and in the prevention and reduction of the symptoms of altitude sickness. This drug is very slightly soluble in water and presents two polymorphic forms with solubility differences. The micronization process (MP) has the potential to increase the water solubility of drugs for bioavailability enhancement. Design of Experiments (DOE) is a beneficial tool in determining the critical process parameters. The aim of this study was to evaluate the MP using DOE to definition of design space for ACZ particle size reduction.Material and Methods: Micronization trials were performed in a Fluid Jet Mill J-20 Micronizer considering feed rate, milling and Venturi pressures as independent variables. Samples were characterized by particle size distribution (laser diffraction), morphology (Image.J) and solubility (shake flask method) were evaluated. The possible crystal structure modification was determined by differential scanning calorimetry (DSC) and X-ray diffraction (XRD). Results and Conclusions: The mean size for ACZ (raw material) was 57.03 ± 0.08µm and for all the micronized samples were less than 6.0µm. The statistical analysis revealed the significant influence provided by milling and venturi pressures on drug particle size distribution. Changes in DSC curves and XRD patterns were observed in the crystalline structure of ACZ and the shake flask study for micronized samples revealed increased solubility in phosphate buffer pH 6.8. This study demonstrates the need of the scientific understanding of the rational selection of the critical micronization process parameters.Financing: CNPq


FCF107-2018RHEOLOGY OF CHOCOLATE DRINK POWDER DEVELOPED WITH RHEOLOGICAL MODIFIER

INGRYD CAROLINNE COSTA CAMPOS(D); SUZANA CAETANO DA SILVA LANNES


Introduction and Objectives: The objective of this work was to evaluate the rheological quality of chocolates drink powder produced with two types of rheological modifiers (guar gum and pregelatinized starch).Material and Methods: The rheological parameters were obtained in a rotational rheometer, Rheotest RN 3.1 Germany, was used S1 concentric cylinder sensor, controlled rate (CR-controlled rate) of 500 1/s in 60 s for the chocolate drink powder with pregelatinized starch at 10ºC and for chocolate drink powder with guar gum it was controlled rate of 250 1/s in 60 s . For the temperature of 60ºC were used the controlled rate parameters 800 1/s in 60s for the chocolate with starch and controlled rate 400 1/s in 60 s for the chocolatado with guar gum. Chocolate drink powder was added in diluted whole milk powder in two different temperatures of consumption (10 and 60ºC) and rheological analysis was carried out. Results and Conclusions: The formulated chocolates drink powder, at both temperatures, adapted to all the rheological models tested. The chocolate drink powder with guar gum showed higher yield value and higher viscosity (0,173 Pa.s for 10ºC and 0,101 Pa.s for 60ºC) when compared to the product with pregelatinized starch (0,051 Pa.s for 10ªC and 0,019 Pa.s for 60ºC), because guar gum has a greater capacity to thicken than pregelatinized starch. The chocolate drink powder with starch (2,723 Pa for 10ºC and 0,406 Pa for 60ºC) presented low initial tension when compared to the chocolate drink powder with guar gum,(6,107 Pa for 10ºC and 0,429 Pa for 60ºC) a result that corroborates with that presented in the viscosity of the products. All the analyzed samples presented pseudoplasticity (n<1). Therefore, the chocolates drink powder with rheological modifier presented good sensorial quality, which is a more viscous product.Financing: CAPES

FCF108-2018DETECTION OF SHIGA TOXIN-PRODUCING Escherichia coli (STEC) CARRING stx2 AND eae GENES ISOLATED FROM CHICKEN MEAT IN DIFFERENT REGIONS OF BRAZIL

ANDRESSA MEM (M), MARIZA LANDGRAF

Food Research Center, Department of Food and Experimental Nutrition, Faculty of Pharmaceutical Sciences, University of Sao Paulo, Sao Paulo, SP, Brazil

Introduction and Objectives: Shiga toxin -producing E. coli (STEC) is one of the most important foodborne pathogens involved in outbreaks, being frequently reported in food of animal origin. Although considered of great importance to public health, there are insufficient reports regarding the presence of STEC in chicken meat in Brazil, the first exporter country. Therefore, it is important to investigate chicken meat samples from poultry slaughterhouse in Brazil. Material and Methods: Eighty-eight chicken meat samples collected from slaughterhouses located in Southern, Southeastern and Midwestern regions of Brazil were analyzed in order to monitor the presence of Shiga toxin-producing E. coli by using Multiplex-PCR according to ISO/TS13136 and USDA MLG 5B.05 methods, both with modifications. Results and Conclusions: Among the Eighty-eight chicken meat samples analyzed for STEC strains, four was positive for Shiga toxin-producing genes stx2 and thirty-three were positive only for eae. Our study detected STEC strains carrying stx2 and eae genes in chicken meat samples, indicating a risk of infection due to the consumption of these products, when it is not properly cooked, or even through cross-contamination. Nevertheless, more research is required with a larger number of samples, since there are few studies regarding STEC in poultry meat reported in Brazil. Financing: Fapesp; CNPq


FCF109-2018STRUCTURE-FUNCTION RELANTIONSHIP BETWEEN DIETARY FIBER FROM PAPAYAS AND THEIR EFFECTS ON COLON CANCER DEVELOPMENT: IN VITRO AND IN VIVO APPROACHES

SAMIRA BERNARDINO RAMOS DO PRADO (D), RODRIGO INVERNORT TAMAROSSI (IC), RENATO HEIDOR, FERNANDO SALVADOR MORENO, JOÃO PAULO FABI

Department of Food Science and Experimental Nutrition, School of Pharmaceutical Sciences, USP

Introduction and Objectives: There is a strong association between dietary fiber (DF) consumption and the reduced risk of colon cancer (CC). Recently, we shown that the DF structure from papaya pulp is strongly affected by fruit ripening and these differences influence their effects on CC cells. Denoting a structure-function relationship is highly desirable for predicting the possible health benefits of polysaccharides and the structural key features of papayas DF is still elusive, though. Thus, we investigated (1) the structure-function relationship between papayas DF and the effects in CC cell lines, and (2) whether the consumption of DF from unripe and ripe papayas has anticancer effects in chemically induced rats for CC development. Material and Methods: Firstly, acidic and neutral DF fractions from papaya were separated by ion chromatography, characterized, and evaluated for effects on distinct CC human cell lines. Then, pre-neoplasic lesions were chemically induced in rats, and it was evaluated whether the consumption of DF from unripe and ripe papaya influences CC development. Results and Conclusions: Results showed that the anticancer effects were dependent of both acidic and neutral DF. Treatment with DF from unripe and ripe papaya changed CC development, in which rats fed with DF from ripe papayas decreased CC foci. Therefore, this study expands the current understanding of the potential health benefits of papaya fruit consumption as a protective agent against CC development. Furthermore, this information may be useful in the exploration of DF from papayas as functional food ingredients.Financing: CNPq; FAPESP.

FCF110-2018STRUCTURAL CHANGES OF VALSARTAN RAW MATERIAL UNDER CONTROLLED ATMOSPHERE

ROSMERY MERMA LEON (M), VINICIUS DANILO NONATO BEZZON (PD), HUMBERTO GOMES FERRAZ


Introduction and Objectives: Valsartan (VAL) is a selective antagonist of the angiotensin II and it is classified as a class II compound according to Biopharmaceutical Classification System (BCS). Its raw material is usually marketed as amorphous form, showing some degree of crystallinity. Usually, amorphous materials, as metastable form, crystallizes into more stable crystalline polymorph when exposed to ambient conditions, however, two amorphous state may also exist in a single component material. Considering the low solubility of VAL and the impact of crystalline transitions on physicochemical properties of drugs, this work aimed to evaluate the stability of the VAL raw material (marketed) under humidity and temperature-controlled conditions, using the X-ray powder diffraction to analyze the changes of the structure.Material and Methods: The powder was kept in a petri dish in a controlled atmosphere (CA) with 75% of humidity and at 40 °C, using a Nova Etica climatic chamber. The X-ray assays were performed using a Bruker D8 Advance diffractometer, operating in a reflection mode, taking a sample every each 10 days.Results and Conclusions: The results showed that the sample under CA, did not crystallize to more stable crystalline form, however, presented a different structure order from that initially observed. It was evident that leaving VAL in conditions of higher atomic mobility, i.e., high humidity and temperature, the disorder of the initial clusters occurred, resulting in the same structural arrangement as amorphous obtained by melt-quenching (MQ), already reported in literature. This study was important to evaluate possible structure transitions of VAL when it is exposed to CA, since these transitions can affect multiple properties of the active ingredients, the most important being the solubility and dissolution rate.Financing: CNPq


FCF111-2018CATALASE-LOADED POLYMERSOMES AS PROMISSING INGREDIENT FOR TOPICAL FORMULATIONS

CAMILA AREIAS DE OLIVEIRA1(PD); CAMILA FORSTER1 (IC); VALKER FEITOSA1 (D); ANDRÉ ROLIM BABY2; PATRÍCIA LÉO3 AND CARLOTA OLIVEIRA RANGEL-YAGUI1

1Department of Biochemical and Pharmaceutical Technology - University of São Paulo, Brazil; 2Department of Pharmacy - University of São Paulo, Brazil; 3Bionanomanufacture Nucleus - IPT, São Paulo, SP, Brazil

Introduction and Objectives: Topical application of catalase is a promising strategy for skin damage induced by UV radiation, including photocarcinogenesis. We experimentally investigated the impact of stirring speed (SS =750 and 1500 rpm); speed time (ST = 24 and 48h) and concentration of catalase (CAT = 0.48 and 0.24 mg/mL) on the safety and efficacy of EO5PO68EO5 polymersomes (polymeric nanovesicles) for topical application. Material and Methods: Based on QbD principles a DoE was applied to develop 8 formulations.DLS and TEM was used to analyze size and morphology of the Ps. The %EE was estimated by direct method. Safety and efficacy were analyzed by HET-CAT, two-dimensional cell models and in vitro skin permeation.Results and Conclusions: The Design space counter plot of hydrodynamic diameter (200 = HD = 400 nm), encapsulation efficiency (2= %EE = 5), polydispersity index (0.1 = HD = 0.3), as a function of ST and CAT indicated that polymersomes with optimized characteristics can be devoped with 0.24 = CAT = 0.40; 40 = ST = 48 and SS= 800 rpm. Encapsulation efficiency (%EE) was determined directly by size exclusion chromatography purification of the polymersomes, with a maximum %EE of 4.72 ± 0.07. Pre-clinical safety tests of HET-CAM and two-dimensional cell models were carried out and the former proved to be more adequate to predict the irritant potential of polymersomes. The skin permeation assay associated with confocal laser microscopy indicated that PS increased over ten times the delivery of loaded molecules to a deeper layer of the epidermis.Financing: FAPESP (2016/03887-4)

FCF112-2018AQUEOUS BIPHASIC SYSTEMS ON THE MULTISTEP PURIFICATION OF CYTOCHROME C PEGYLATED FORMS

JOÃO HENRIQUE PICADO MADALENA SANTOS (D)*,**;GUSTAVO CARRETERO (PD)***, SÓNIA PATRÍCIA MARQUES VENTURA (PD)*; CARLOTA OLIVEIRA YAGUI- RANGEL (PD)**

*CICECO, Departamento de Química, Universidade de Aveiro, Portugal. ** Department of Biochemical and Pharmaceutical Technology, SP Brazil. ***Institute of Chemistry, São Paulo University, SP Brazil

Introduction and Objectives: The physicochemical properties and kinetics of proteins can be improved by chemical PEGylation. However, usually a mixture of heterogeneous PEGylated conjugates and unreacted protein is obtained, challenging the design of an efficient downstream process. The purification of PEGylated proteins normally addresses (i) the separation of PEGylated conjugates from the unreacted protein and (ii) the fractionation of the PEGylated conjugates considering their degree of PEGylation. In this work, a process was proposed to purify PEGylated cytochrome c (Cyt-c) by aqueous biphasic systems (ABS). Material and Methods: Firstly, the partition behavior of Cyt-c c and its PEGylated conjugates (Cyt-c-PEG-4 and Cyt-c-PEG-8) on polyethylene-glycol (PEG) + potassium phosphate buffer (pH = 7) aqueous biphasic systems (ABS) was investigated. Circular dichroism was used to investigate possible protein denaturation during PEGylation and/or purification. Results and Conclusions: PEGs of intermediate molecular weight allowed separation of the PEGylated conjugates from the unreacted protein in a single step. Following, efficient separation of the different PEGylated forms was attained using ABS formed by PEGs of high molecular weight. Additionally, unreacted Cit-c was successfully recycled. Protein stability after purification was confirmed by circular dichroism. A downstream process to separate Cyt-c, Cyt-c-PEG-4 and Cyt-c-PEG-8 with high purities (96.5%, 85.8%, and 99.0%, respectively) was developed.Financing: FAPESP and the FCT.


FCF113-2018A UNIQUE ORAL SUPRAGENGIVAL PLAQUE MICROBIOME OF A REMOTE BRAZILIAN RIVERINE POPULATION AND ITS CORRELATION TO THEIR DIET

POLIANA SCARCELLA D’OLIVEIRA HERSZKOWICZ (IC), PEDRO JOSÉ TÓTORA DA GLORIA (PD), FABIANA ANDREA HOFFMANN-SARDA (PD), FABIANA DA SILVA LIMA (PG), RODRIGO ELIAS DE OLIVEIRA (PD), MARIA REGINA LORENZETTI SIMIONATO, CHRISTIAN HOFFMANN


Introduction and Objectives: We present data on the supragingival plaque microbiome of a remote riverine population living in the Amazon, as a biocultural model to aid our understanding of traditional populations’ oral health. The aim of this work was to evaluate the effects of the riverine population’s diet on the oral microbiota, and their oral health. Material and Methods: Occurrence of oral pathologies, diet intake and overall health parameters were accessed, and supragingival plaque was collected from 113 individuals. DNA was extracted and microbial profiling was done using the bacterial 16S rDNA. Sequence analysis was carried out using Qiime, and statistical analysis was carried out using the R environment for statistical computing. Results and Conclusions: Dental caries were detected in 88,5% of the subjects and 99% of the subjects had at least one decayed, missing or filled teeth. The supragingival plaque microbiome showed 150 different genera, detected in at least 2 subjects. Permanova analysis indicated a global relationship between the oral microbiome and: the proportion of tooth loss (unweighted unifrac pvalue: 0.046), the frequency of sugar consumption (unweighted unifrac pvalue: 0.024), and the proportion of filled cavities (pvalue=0.005 and 0.045, for unweighted and weighted unifrac). Tooth brushing frequency, flossing and smoking were not related to microbiome composition. A unique oral microbiome signature was detected and further analysis is underway to clarify the relationships between this population and ancient and modern populations. Financing: FAPESP; Capes

FCF114-2018MICROBIOLOGICAL SAFETY INDICATORS OF CANASTRA CHEESE IN BRAZIL

GABRIELA ZAMPIERI CAMPOS (M), BERNADETTE DORA GOMBOSSY DE MELO FRANCO, CHRISTIAN HOFFMANN, GUSTAVO AUGUSTO LACORTE, MARIZA LANDGRAF, UELINTON MANOEL PINTO

FoRC, Department of Food and Experimental Nutrition, Faculty of Pharmaceutical Sciences, University of Sao Paulo, SP, Brazil

Introduction and Objectives: Canastra Artisanal Minas Cheese is made from raw milk by small farmers in Serra da Canastra region. The production process employs fermentation by an endogenous culture called “pingo” originated from the whey collected from the previous day’s production. The use of raw milk is a risk factor for food safety and potential foodborne pathogens should be controlled during the 22 days ripening period required by law. This work analyzed the microbiological safety indicators of Canastra Cheese from 78 rural properties. Material and Methods: Total coliform, Escherichia coli and Staphylococcus coagulase positive counts were performed on Petrifilm® plates (3M). The detection of Salmonella spp. and Listeria monocytogenes was performed by using ISO6579:2002 and ISO11290-1:1996/(A)1:2004 methods. Enterobacteraceae counts were determined by the APHA method 9.62:2015. The pH analyzes were performed according to IAL 463-IV method and the Water Activity(aW) in an AquaLab analyzer. Results and Conclusions: The results show that 62% of samples were unsatisfactory to least one requirement of the legislation. pH and aW did not significantly influence whether the samples would be classified as satisfactory according to the legislation (p <0.05), suggesting no influence of these parameters on the microbiological indicators. No sample was contaminated with Salmonella and L. monocytogenes was found in one sample. The high number of non-compliances and the high counts of E. coli or total coliforms or Staphylococcus in some samples indicate a need to implement or improve hygienic-sanitary conditions in the production of Canastra cheese. Financing: Fapesp (Proc. 2013/07914-8) and Capes


FCF115-2018STANDARDIZATION OF SOYBEAN MEAL COUNTING DRY-HEAT STERILIZED

LUANA CRISTINA VAZ FORMAGIO (IC) CLEIDE MARIA DE OLIVEIRA AMARAL (CONFAR) MARINA DE SOUZA BRAGA (PD) TEREZINHA DE JESUS ANDREOLI PINTO

Department of Pharmacy, Faculty of Pharmaceutical Sciences, University of São Paulo

Introduction and Objectives: The aim of this work was to prepare the inoculum of Salmonella typhimurium (ATCC 14028), its standardization and counting through methodology described in standard operating procedure available in the microbiological laboratory - CONFAR in soybean meal sterilized by dry-heat.Material and Methods: Salmonella typhimurium was grown in TSA ( Soy tryptone agar) culture medium incubating for ± 24h / 30-35 °C. 5 mL of 0.9% Sodium Chloride was added to remove the microbial grown, then homogenized to recover all microbial growth. After this step, a serial dilution of 10-1 to 10-8 was performed. At the end 1mL of the last 3 dilutions (triplicate) were plated in TSA medium and the plates incubated for ± 24 h / 30-35 °C. To do the count in the sample, 10 g were weighed under aseptic conditions (triplicate and 1 sample control) and resuspended in 90 mL of TSB (Trypticase Soy Broth), the vials were contaminated with the pre- standardized inoculum. A further serial dilution was done and 1 mL of each plate dilution (triplicate) in TSA medium and counting was performed after incubation. The samples of soybean meal were applied in triplicates. Results and Conclusions: It is concluded that the results of this standardization were satisfactory, positive control presented growth and showed consistency between the final results in triplicates.The sample used did not present the microorganism Salmonella typhimurium. Financing: FIPFARMA

FCF116-2018EVALUATION OF CYTOTOXICITY IN BREAST CANCER CELL LINES DERIVED FROM NANOTECHNOLOGICAL IDARUBICIN FORMULATIONS

ROSANGELA MENDES CARVALHO DOS SANTOS (IC) *, ** MARINA DE SOUZA BRAGA (D)*,** DANIELA DAL MOLIM GHISLENI (D)* PATRICIA LÉO (D)*** ADRIANO MARIM DE OLIVEIRA (D)*** NATALIA NETO PEREIRA CERIZE (D)***

* Department of Pharmacy, Faculty of Pharmaceutical Sciences, University of Sao Paulo ** CONFAR Laboratory *** Nucleus of Bionanomanufatura, Institute of Technological Research, Sao Paulo

Introduction and Objectives: The aim of this work was to evaluate citotoxicity and compare different formulations of idarubicin encapsulated in nanoparticles and in lipossomal formulation with idarubicin in free solution. Material and Methods: An aqueous solution of Idarubicin was mixed with poly(dimethylsiloxane)-graft-polyacrylates (0.1% w/v) and pH was adjusted to 8.2 for solubilization. The sample was spray-dried using an atomization membrane with 4 &mum of porosity. Unilamellar liposomes were prepared by the extrusion method, were used DMPC and DSPE-2000 (95:5 mol%). Cytotoxicity assay was performed after standardized exposure time of 24 hours and was determined by the coloration with neutral red. In all experiments the positive control of cytotoxicity with Dodecius sodium sulfate was used.Results and Conclusions: The results show that there was no significant difference between the concentration of free Idarrubicina and the formulations studied in relation to IC50, in the most aggressive cell line T47D needed a greater concentration ~ 0.01 mg/ML to reach 50% of death, whereas in the less aggressive cell line MCF-7 was ~ 0.006 mg/ML. These results showed that, despite the drug encapsulation process there is no need of increase doses to achieve IC50 and the results of both tested formulations associated with other results achieved have demonstrated to be promising in the treatment of breast cancer in order to decrease toxicity. Financing: FAPESP


FCF117-2018EFFECTS OF HYDROQUINONE EXPOSURE ON THE ADAPTATIVE IMMUNE RESPONSE INDUCED BY INFLUENZA VACCINE

ANDRÉ LUIS FABRIS (M); GUSTAVO HENRIQUE OLIVEIRA DA ROCHA (D); MARINA DE PAULA-SILVA (D); EDUARDO LANI VOLPE DA SILVEIRA (PD); SANDRA HELENA POLISELLI FARSKY

School of Pharmaceutical Sciences, University of São Paulo, São Paulo, SP, Brazil

Introduction and Objectives: Influenza is a health problem worldwide responsible for causing severe complications. Vaccination is the best way to prevent and protect against it. On the other hand, evidence has supported that immune responses could be impaired by exposure to environmental pollutants, such as those found in cigarette smoke (CS). We have already shown that hydroquinone (HQ), a constituent of CS, leads to several alterations on the immune system. We sought to investigate the effects of in vivo HQ exposure on the adaptive immunity induced by influenza vaccine. Material and Methods: Male C57BL/6 mice were daily exposed to HQ (2500 ppm) or PBS for 8 weeks (1h/day). At weeks 6 and 8, mice were immunized with the influenza vaccine. Samples were collected at weeks 6 and 8 during HQ exposure and 7, 35 and 70 days after the vaccine booster. CEUA/FCF protocol #561.Results and Conclusions: HQ exposure caused a slight reduction in erythrocytic parameters and no changes on body weight or of biochemical markers of liver and kidney functions. However, it increased the oxidative stress in splenocytes, indicating a potential mechanism of HQ toxicity. Also, those animals presented larger lymph nodes, lower frequency of antibody-secreting cells and altered morphology of spleen follicles. Whereas avidity and titers of specific IgG were not modified, IgG2c titers of HQ exposed-animals were reduced though, suggesting a diminished maintainance of humoral response. Thus, HQ exposure caused no apparent toxicity, albeit immune toxic effects were detected after immunization. Financing: Fundação de Amparo à Pesquisa do Estado de São Paulo – FAPESP (Process nº 2016/24134-4)

FCF118-2018COULD AYAHUASCA SUPRESS THE ACUTE EFFECTS OF COCAINE?

JÉSSICA RUMI KOIAMA1 (IC), VITOR BRUNO1 (M), VINÍCIUS RODRIGUES NEUGEBAUER1 (IC), ROSANA CAMARINI2, TANIA MARCOURAKIS1

1Department of Clinical and Toxicological Analyses, School of Pharmaceutical Sciences, University of São Paulo; 2Department of Pharmacology, Institute of Biomedical Sciences, University of São Paulo

Introduction and Objectives: Behavioral evaluation of the acute effect of ayahuasca (AYA) as a protective factor to the psychostimulant effects of cocaine. Material and Methods: (CEUA/FCF 33.2016-P518) C57Bl/6 adult mice were habituated in the open field apparatus (OF) for 3 consecutive days and 24h later; they received a dose of water or AYA 1.76; 3.0; 17.6 or 30.0 mg DMT/kg v.o. Locomotor activity of the animals was evaluated for 1h. Then, AYA 3.0mg DMT/kg or saline v.o. was administered and mice were submitted to the OF for 30 min. A challenge with cocaine 10mg/kg or saline i.p was made and they returned to the OF for 30 min.Results and Conclusions: Results and discussion: In the dose-response curve experiment, the AYA-treated group 1.76 mg DMT/kg showed a slight hyperlocomotion compared to the control, and the groups treated with the two higher doses presented a tendency to hypolocomotion. However, animals treated with 3.0 mg DMT/kg showed similar locomotor activity to the control group, that’s why it was the dose chosen for the experiments. Thus, it was observed that the AYA-group showed similar locomotor activity to the control group when they were challenged with cocaine. It’s believed that the dose of AYA containing 3.0 mg DMT/kg is insufficient to acutely inhibit the psychostimulant activity of cocaine since this drug causes robust neuroplastic changes. Conclusion: AYA did not significantly alter the locomotor activity of mice. Moreover, the acute use of AYA containing 3 mg DMT/kg v.o. does not have a protective factor for the neuroplastic changes triggered by cocaine use. Financing: SENAD, FAPESP


FCF119-2018DESIGN AND SYNTHESIS OF TRIAZOLYL-FLAVONE HYBRIDS AND THEIR IMPLICATION IN THE TREATMENT OF LEISHMANIASIS

PONE KAMDEM BONIFACE (PD), ELIZABETH IGNE FERREIRA

Department of Pharmacy, Faculty of Pharmaceutical Sciences, University of São Paulo

Introduction and Objectives: Leishmaniasis is an infectious disease caused by protozoa of the genus Leishmania and is responsible for over 20 000 deaths per year. The treatment of leishmaniasis relies primarily upon chemotherapeutic drugs like sodium stibogluconate, amphotericin B, pentamidine and most of these drugs cause severe side effects. Therefore, there is an urgent need to search for new and safe antileishmanial potential leads to treat leishmaniasis. The aim of present study was to design and synthesize some flavone derivatives with the purpose of obtaining potentially active compounds against leishmaniasis. Material and Methods: A series of chemical reactions including the preparation of different azides, the methylation of appropriate acetophenone derivatives, the propargylation of 3,4- dihydroxybenzaldehyde; the protection of different -OH groups by methylation, the preparation of appropriate chalcones, and the synthesis of the triazole compounds through click-chemistry using the prepared chalco-alkynes and azides was carried out. Results and Conclusions: A panel of seven triazolyl amalgamated flavones was synthesized with good to excellent yields (25-99 %). The compounds were characterized by 1H NMR and 13C NMR. The evaluation of the antileishmanial activity of these compounds is the subject of further introspection.Financing: FAPESP

FCF120-2018STABILITY STUDY OF THE CASEIN SOY AGAR AND THIOGLYCOLLATE CULTURE MEDIUM USED IN THE CONFAR MICROBIOLOGICAL LABORATORY

ELIZIÁRIA INÁCIO DE ABREU LIMA (IC)*, ** GABRIELA CORRÊA CARVALHO (M) * MARINA DE SOUZA BRAGA (PD) *, ** CLEIDE MARIA DE OLIVEIRA AMARAL** TEREZINHA DE JESUS ANDREOLI PINTO*, **

* Department of Pharmacy, Faculty of Pharmaceutical Sciences, University of Sao Paulo, Sao Paulo ** CONFAR Laboratory

Introduction and Objectives: The present study aims to validate reference culture media used in the Laboratory of Control of Medicines, Cosmetics, Domains, Related Products and Raw Materials (CONFAR).Material and Methods: Both culture media (casein soy agar and thioglycollate) were prepared according to the manufacturer’s guidelines in two different volumes, 70 mL and 100 mL with two lid types, gauze and hydrophobic cotton and polymeric screw lid respectively. Both were analyzed for volume and pH after 24 hours of preparation (T0), 60 days (T1), 90 days (T2) and 120 days (T3). In these same times the growth promoter capacity was analyzed using S. aureus, P. aeruginosa and C. perfringes for thioglycollate, whereas C. albicans, A. brasiliensis and B. subtilis used for casein soy agar. Results and Conclusions: Results obtained for pH in thioglycollate media were maintained according to the manufacturer’s specifications, unlike the casein soy agar with deviation starting in T3, regardless of the lid type fell below the specified. In both culture media the volume decrease was directly proportional to the storage time but lower in polymeric screw lid. Both culture media presented growth promoter capacity at all times analyzed. In T3 the thioglycollate reddish disk was not anymore observed; in addition, there was a change in the medium color that may be indicative of degradation. Even so, due to the decrease in volume and consequent concentration of agar and lower diffusion of oxygen, the development of C. perfringes, anaerobic bacteria in both lids. Both analyzed culture media remained stable until the period of 120 days.Financing:


FCF121-2018IDENTIFICATION OF TWO ISOMERS OF CONJUGATED LINOLENIC ACIDS IN WISTAR FECES

GIOVANNA CAVANHA CORSI (IC), LUCIANA TEDESCO YOSHIME (PD), ILLANA LOUISE PEREIRA DE MELO (PD), JOSÉ AUGUSTO GASPAROTTO SATTLER (PG), JORGE MANCINI-FILHO

Department of Food Science and Experimental Nutrition, FCF/USP

Introduction and Objectives: Conjugated linolenic acids (CLNAs) alter lipid metabolism, has anti-carcinogenic and immunomodulatory properties. However, CLNAs can be metabolized to conjugated linoleic acids (CLAs) and their beneficial properties may be due to these compounds. Studies with CLAs are controversial and some kind of health-related side effects. The aim of this study was to evaluate the fatty acids (FA) composition in the feces of animals supplemented with CLNA from pomegranate (PSO) and bitter gourd seed oil (BSO). Material and Methods: The study was approved by the local ethics committee (CEUA – no. 433, 436). The Wistar rats were treatment for 40 days, divided in four groups: Control (received water), CLA, PSO and BSO. The supplementation was intragastric and the percentage offered was in relation to daily feed intake. Feces were collected in three different times (t0=initial, t1=20 and t2=40 days). The lipid extraction was Folch method followed by methylation, and the identification and quantification of the FA were by Gas Chromatography. Results and Conclusions: All treated groups have the FA of the Control group (C16:0 and C18:2c n6). PSO has high concentrations of punicic acid (PA) and BSO, a-eleostearic acid (EA). The results of the supplementation confirm the presence of the PA (t0=0; t1=2.6±0.2; t2=2.4±0.3), EA (t0=0; t1=1.1±0.1, t2=0.6±0.1), and CLAs (C18:2 9c11t - t0=0; t1=1.1±0.1; t2=1.7±0.3 and C18:2 10t11c - t0=0; t1=1.4±0.1; t2=1.7±0.3) on feces. Some studies indicated that PA and EA are absorbed in rat intestine, and a portion is converted to CLA. Interestingly, CLAs were found only in the supplemented group. Therefore, once the safety of PSO and MSO is confirmed, they may have value as health food supplements.Financing: CNPq, FAPESP

FCF122-2018SYNTHESIS OF DENDRIMERIC NANOCARRIER OF INTRACELLULAR RELEASE: TRIPANOMICIDAL DENDRIMER PRODRUG CONTAINING 3- HYDROXYFLAVONE

RODRIGO VIEIRA GONZAGA (D), LUCAS ADRIANO DO NASCIMENTO (IC), ELIZABETH IGNE FERREIRA

LAPEN, Department of Pharmacy, FCF-USP

Introduction and Objectives: Several strategies have been used to improve drug candidates and even those drugs already on the market. Nanotechnology is a promising field in the pharmaceutical field and widely used in the designing of nanosystems for drug delivery associated to prodrug design strategies. This universe encompasses the “stimuli-responsive” and “self- immolative” intelligent transporters capable of triggering the release of bioactive agents from the carrier, such as: drugs, genes and proteins, in response to specific cellular signals. Neglected diseases (ND) are known as poverty-promoting diseases, caused by socioeconomic inequalities and promoting such inequalities. Given the current problem in the treatment of ND and the great need and urgency of new drugs in therapeutics, this project aims to develop a new system for delivery and intracellular release sensitive to glutathione reductase / trypanothione reductase (GSH / TR) by a disulfide crosslinker promoting the cascade self-release of the bioactive compound, capable of improving pharmacokinetic properties of 3-hydroxyflavone. Material and Methods: The dendrimeric nanocarrier synthesis is divided into two blocks. The first one is a click chemistry reaction-based dendron, which comprises the use of the propargilated and azide versions of trometamol. This dendron and the drug are linked to a disulfide crosslinker through carbamate bonds.Results and Conclusions: All compounds were characterized by NMR 1H and 13C, infrared spectrophotometry and melting point measurements. Financing: CNPq; CAPES.


FCF123-2018CIGARETTE SMOKE EXPOSURE DURING PREGNANCY INCREASES THE SUSCEPTIBILITY OF THE INFLAMMATION PROCESS IN THE CENTRAL NERVOUS SYSTEM (CNS) OF THE OFFSPRING

ANA CAROLINA DURÃO1 (D), WESLEY BRANDÃO2 (PD), NÁGELA GHABDAN2 (D), JEAN PIERRE PERON2, TANIA MARCOURAKIS1

1Faculty of Pharmaceutical Sciences, University of São Paulo - Department of Clinical and Toxicological Analysis - Pharmacy (Physiopathology and Toxicology) Program, 2Institute of Biomedical Sciences, University of São Paulo - Department of Immunology

Introduction and Objectives: During the implantation phase, the embryo is more vulnerable to external influences such as cigarette smoke, which can increase the risk of fetal developmental delay and immune system abnormalities. This study evaluated the effect of cigarette smoke exposure during pregnancy on an inflammatory response in the CNS of the offspring. Material and Methods: C57BL/6 mice were exposed to 3R4F cigarette smoke, or synthetic air, from vaginal plug to offspring birth. At the 3rd day of life, offspring were separated for the following studies: 1) in vitro: brains were dissected and a mixed glial culture was prepared. After 21 days, the cells were stimulated with 100ng/mL LPS or culture medium. After 24h, the pro-and anti- inflammatory cytokines were evaluated by CBA, as well as GFAP, CD11b, CD80 and CD86 by flow cytometry. 2)in vivo: animals were challenged with LPS (1mg/kg) or saline ip. After 4h, the mice were euthanized and the CNS removed for PCR analysis. Results and Conclusions: The in vitro experiments showed a proliferation of astrocytes and an increase in proinflammatory cytokines in the cells of FPS group when compared to the CO. In vivo the PCR showed that mice exposed to cigarette smoke and LPS had decreased expression of the CCR2 gene. Our results suggest that exposure to cigarette smoke during pregnancy increases the inflammation response in the CNS of the offspring after an inflammatory stimulus. Financing: CAPS; CNPq; FAPESP

FCF124-2018ANTICHAGASIC DRUG CANDIDATES: DESIGN AND SYNTHESIS OF INTRACELLULAR RELEASING DENDRON NANOCARRIER AND BIOACTIVE COMPOUNDS

LUCAS ADRIANO DO NASCIMENTO (IC), RODRIGO VIEIRA GONZAGA (D), ELIZABETH IGNE FERREIRA

LAPEN, Department of Pharmacy, FCF-USP

Introduction and Objectives: In view of the urgent need of drugs for neglected diseases, including Chagas disease, and taking into account the importance of dendrimers on the improvement of bioactive compounds, this research aims to synthesize dendrons – part of the dendrimers – of trometamol to be used as carrier for 3-hydroxyflavone, an in vitro bioactive compound without in vivo activity. With the purpose of obtaining trypanomicide dendrimeric candidates derived from this flavone, glutathione reductase/trypanothione reductase sensible intracellular drug releasing systems were designed. This derivative could be an important background for the construction of intelligent nanocarriers, with self-immolative properties.Material and Methods: Using the convergent approach, the synthesis of the dendrons and the dendrimer as well were based on introducing tryazol linkers by means of “click chemistry. The azides of trometamol react with propargilated trometamol. The obtained compounds were characterized by 1H NMR and 13C NMR, Infrared and melting point determination. Results and Conclusions: The intermediate trometamol-BOC was obtained with a 70% yield, the intermediate azides, with a 91% yield, and the propargyl-trometamol intermediate, with a 20% yiled. Financing: CNPq, CAPES


FCF125-2018DEVELOPMENT OF POLYMERSOMES FOR THE ENCAPSULATION OF L-ASPARAGINASE

AMANDA TURNO DA SILVA (IC), ALEXSANDRA CONCEIÇÃO APOLINÁRIO (D), ADALBERTO PESSOA JUNIOR, CARLOTA DE OLIVEIRA RANGEL YAGUI


Introduction and Objectives: L-asparaginase (ASNase) is one of the main chemotherapeutic agents for the therapy of acute lymphoblastic leukemia (ALL). The use of nanosystems aiming the delivery of this protein is an approach to avoid its immediate immunogenic effects and to increase half-life. Polymersomes (PL), vesicles formed from amphiphilic block copolymers, are an interesting strategy used for protein drug delivery. Therefore, the production of PL and their capacity to encapsulate ASNase were investigated.Material and Methods: PL of poly(ethylene glycol) poly(lactic acid) (PEG2000-PLA5000, PEG5000-PLA10000 and PEG5000-PLA11000) were prepared by polymeric film hydration (FH). The copolymers were dissolved in chloroform (1mg.mL-¹) and the solvent was evaporated. The film obtained was hydrated with PBS pH 7.4. PL of Pluronic L-121 (PEO5- PPO68-PEO5) were prepared by temperature switch method. The PL size was determined by DLS and morphology by TEM. Results and Conclusions: For PEG-PLA after the initial FH, high polydispersity index (PDI) values were found (PDI > 0.4). Nonetheless, centrifugation and extrusion were able to narrow the PL PDI for all PEG-PLA copolymers tested to values under 0.4. Encapsulation efficiency of ASNase by indirect method was 25%, 15% and 8% for PEG2000-PLA5000, PEG5000- PLA10000 and PEG5000-PLA11000, respectively. For Pluronic L-121, PDI values without extrusion were approximately 0.1 and ASNase encapsulation efficiency was close to 5%. All PL were about 100-300 nm in size. The main challenges to obtain PL and to encapsulate ASNase are related to the vesicles production, control of their sizes and PDI values.Financing: FAPESP; CNPq.

FCF126-2018DESIGN AND CHARACTERIZATION OF FLOATING TABLETS: A COMPARISON BETWEEN EFFERVESCENT AND NON-EFFERVESCENT APPROACHES

JESSICA LINS DOS SANTOS (IC), AMANDA CAMPOS FORTES (D), MICHELE GEORGES ISSA (PD), HUMBERTO GOMES FERRAZ


Introduction and Objectives: Floating Drug Delivery Systems are classified as effervescent and non-effervescent depending on the buoyance approach employed. These systems benefit drugs that have the upper gastrointestinal tract as preferential site for their absorption and/or site for their pharmacological activity and have significantly high solubility on acidic medium. This study aimed to design floating tablets and to evaluate the impact of floating agent and processing conditions on drug release, swelling and buoyancy.Material and Methods: The impact of floating agent (effervescent and non-effervescent) and processing conditions (direct compression, granulation and hardness) on drug release, buoyancy and swelling were evaluated following a 23 factorial design. HPMC K4M was used as the release matrix, cellulose as a diluent agent, a low-density polypropylene foam powder and sodium bicarbonate were used as non-effervescent and effervescent floating agents, respectively. Results and Conclusions: Drug release was higher for non-effervescent tablets obtained by wet granulation method. Tablets with hardness of 40N showed an increase in floating time, geometric density, porosity and friability when compared with tablets with hardness of 80N. Effervescent tablets obtained by wet granulation showed higher swelling, as non- effervescent tablets with hardness of 80N. This study showed that the drug release modulation of floating tablets is directly dependent of combination of floating agent approach, the tablet process obtention and hardness. Financing: DEINFAR/FCF/USP


FCF127-2018PROFILE EVALUATION OF PHARMACY GRADUATES FROM SCHOOL OF PHARMACEUTICAL SCIENCES OF THE UNIVERSITY OF SÃO PAULO

PALOMA CLEMENTINO NASCIMENTO (IC); WALDIR CARLOS DOS SANTOS (IC); ELVIRA MARIA GUERRA-SHINOHARA

Department of Clinical and Toxicological Analyses, FCF/USP

Introduction and Objectives: The purpose this study was to evaluate academic and professional characteristics of Pharmacy-Biochemistry graduates from School of Pharmaceutical Sciences of the University of São Paulo. Material and Methods: A form was made containing the following information: student’s age at admission, semester graduation, internships (Scientific Initiation, Pharmaceutical Activities, Attention or Pharmaceutical Practices, internships abroad), participation in complementary activities, questions about graduate employment and professional pharmaceutical area. This form was available on school page and collected with the documents for collation of the degree. Results and Conclusions: At this moment, 201 graduates have been included, of which 88 were in the full period course (I) and 113 in the night course (N). It was observed a small frequency of students who graduated in the ideal time I=5.7%, and N=18.6%. The disciplines that presented the greatest number of disapprovals were: Organic Chemistry I, II and III; Heterocyclic Compounds, Calculus, Analytical Chemistry and Instrumental Analytical Chemistry. The frequencies of graduates who did internships abroad were I=42.0% and N=31.0%. Most of the students did Scientific Initiation (I=70.5% and N=58.4%). The majority of graduates choose the industry for the qualification in Pharmaceutical Activities, in both courses. Only 35.2% of I and 14.2% of N participated in the Scientific Journey, with the participation in the Diabetes and Hypertension Campaign of I=59.1% and N=33.6%. The majority (I=78.4% and N=71.7%) participated at least once in SUPFAB. The vast majority of undergraduates who graduated were employed (I=70.5% of the full period and N=76.1%). Financing: PUB-USP

FCF128-2018SAFETY AND SELECTIVE CYTOTOXICITY OF MILTEFOSINE-LOADED POLYMERIC MICELLES

VALKER ARAUJO FEITOSA*(D), CAMILA AREIAS DE OLIVEIRA*(PD), ANTÔNIO FERNANDO MONTEMOR**, PATRICIA LEO**, NATÁLIA NETO PEREIRA CERIZE**, CARLOTA DE OLIVEIRA RANGEL-YAGUI*

* Department of Biochemical and Pharmaceutical Technology, ** Bionanomanufacturing Center, Institute for Technological Research (IPT)

Introduction and Objectives: Miltefosine (MTF) is a synthetic antitumor alkylphosphocholine used clinically for the treatment of cutaneous metastases of breast cancer. This drug act at cell membrane level, where it accumulate and interfere with lipid metabolism and lipid-dependent signaling pathways leading the cells to apoptosis. As a surface-active substance, higher concentrations of MTF may cause unspecific cell lysis and thus are not suitable for intravenous administration as well as present pronounced gastrointestinal toxicity. To overcome these limitations we incorporated MTF into polymeric micelles (PM).Material and Methods: Drug-loaded PM were prepared by direct hydration technique using Pluronic F127, a biocompatible triblock-copolymer approved by FDA. Results and Conclusions: The MTF incorporation into PM enhanced the safety by reducing in vitro hemolytic potential, around 5-folds, increasing the drug concentration that could be administered intravenously. Then, using a chicken chorioallantoic membrane (CAM) model, we also showed that MTF-PM prevented mucosal irritation, decreasing bleeding and lysis of blood vessels. In addition, we observed a dose-dependent reduction in viability of human tumor cell lines (i.e. HeLa and HepG2) when treated with MTF, while its action is selective as MTF does not influence normal L929 fibroblasts. Interestingly, MTF-PM increased the cytotoxicity against Hela cells, suggesting a differential uptake of these nanostructures by those cells. These data together shown that MTF encapsulation into PM reduces side effects and increases selectivity cytotoxicity against tumor cells. Financing: CAPES; FIPT; CNPq and FAPESP.


FCF129-2018EXPLORING DIETARY FIBERS FROM INDUSTRIAL BY-PRODUCTS: ARABINO-OLIGOSACCHARIDES FROM SUGAR BEET DIFFERENTIALLY REGULATES MACROPHAGE FUNCTION

VICTOR COSTA CASTRO-ALVES (PD); JOÃO ROBERTO OLIVEIRA DO NASCIMENTO

Department of Food Science and Experimental Nutrition, School of Pharmaceutical Sciences; Food Research Center (FoRC), São Paulo Research Foundation

Introduction and Objectives: As the concern for healthy habits is growing, the interest in the sustainable production of dietary components that may promote effects beyond its nutritional function is also increasing. Dietary fiber (DF) from sugar beet is an industrial by-product with great potential to be used in foods as an emulsifier. Furthermore, the enzymatic hydrolysis of this DF produces linear arabino-oligosaccharides (AOS) with varying size (degree of polymerization - DP), which are known for their probiotic effects. Although DF can also regulate intestinal immune function through direct interaction with phagocytes, it is unclear (1) whether AOS regulate phagocytes function and (2) whether the size of AOS influences their potential biological effects. Material and Methods: Four AOS (DP ranging from 3 to 6) prepared from DF of sugar beet were characterized through time-of-flight mass spectrometry and anion-exchange chromatography. Then, enzyme-linked immunosorbent assay, fluorescence, flow cytometry and gene expression analysis were used to evaluate the effects of these AOS in macrophages previously activated or not by lipopolysaccharide.Results and Conclusions: AOS with varying DP differentially regulate the production of pro-inflammatory cytokines and chemokines in activated macrophages. AOS also inhibited LPS-induced reactive oxygen species production. Since LPS-triggered macrophages help to disrupt the intestinal epithelial barrier integrity in response to an unbalanced diet, the induction of AOS with specific size in food and beverages may increase the potential health benefits associated with the consumption of these food products.Financing: FAPESP; PNPD-CAPES

FCF130-2018BENEFICIAL EFFECTS OF THE USE OF BUFFALO MILK IN THE ELABORATION OF PROBIOTIC DAIRY DRINKS

THAMIRES MARIA SIMÕES DA SILVA (D), RICARDO PINHEIRO DE SOUZA OLIVEIRA


Introduction and Objectives: Probiotic dairy drinks are recognized for having in their composition microorganisms with potential benefits to consumer health. The use of buffalo milk to prepare dairy drinks may be a favorable alternative due to its nutritional superiority when compared to bovine milk, mainly in relation to the higher levels of protein and fat, which may influence the technological characteristics of the products. Therefore, this study evaluated buffalo and probiotic dairy drinks as the parameters of post acidification, the viability of the fermentation cultures and their resistance to in vitro gastrointestinal stress. Material and Methods: Six formulations of probiotic dairy drinks were made using buffalo or cow’s milk in different proportions of 0%, 25 and 50% of whey. The milk was fermented to 42 °C until reaching pH 4.7 using S. thermophilus, L. bulgaricus and L. acidophilus and subsequently mixed with whey in appropriate proportions. The milk drinks were stored at 4 °C for 21 days. During this period, milk drinks were evaluated for post-acidification, number of viable bacteria and resistance to gastrointestinal stress in vitro.Results and Conclusions: The buffalo milk drink (50%) showed a better pH stability over the analysis period. The buffalo probiotic dairy drinks indicated the better viability of the fermenter cultures throughout the refrigerated storage and at the end of the gastrointestinal stress simulation. The buffalo probiotic milk drink containing 0% whey had the highest viable cell count in both analyzes. The use of buffalo milk in the elaboration of probiotic dairy drinks presented benefits as the post acidification parameters as to the viability of the fermentation cultures throughout the storage period and after in vitro gastrointestinal simulation.Financing: CNPq; CAPES


FCF131-2018LIPID NANOPARTICLES FOR ITRACONAZOLE CUTANEOUS DELIVERY

JULIA SAPIENZA PASSOS (IC)*, VANESSA FRANCO CARVALHO DARTORA (D)**, LUCIANA BIAGINI LOPES**

* School of Pharmaceutical Sciences of São Paulo, University of São Paulo **Institute of Biomedical Sciences, University of São Paulo, São Paulo

Introduction and Objectives: This study aimed at developing nanostructured lipid carriers (NLC) for topical delivery of itraconazole, aiming the improvement of cutaneous drug localization in the treatment of human and veterinary sporotrichosis lesions. Material and Methods: The nanocarriers were developed combining solid (glyceryl behenate) and liquid (tricaprylin) lipids at ratios ranging from 7:3 to 3:7 (w/w), surfactants and water. They were characterized for size, morphology and zeta potential. Itraconazole was added at final concentrations of 1-5% (w/w). Transepidermal wate¬¬r loss was measured as an index of tissue permeability after skin treatment with nanoparticles or water (control) for 24 h; as model tissue, we employed intact porcine skin or skin damaged with a linear incision to mimic a wound. Results and Conclusions: Ratios of solid:liquid lipid of 5:5 and higher resulted in nanoparticles with diameter smaller than 230 nm (polidispersity index <0.25) and negative zeta potential (-15 to -20 mV). Itraconazole solubilization in the system increased with the liquid lipid content, but drug concentrations higher than 3% resulted in precipitation and increases in particle size and polidispersity (over 2-fold). The nanoparticles decreased transepidermal water loss in the intact skin and in tissues subjected to an incisional wound in 35 and 42%, respectively. These results demonstrate the possibility to obtain itraconazole-loaded lipid carriers within the nanometric range, and the ability of the nanoparticles to preserve skin hydration even in damaged tissue, which is considered a key feature to ensure an efficient healing.Financing: CNPq/PIBIT

FCF132-2018MOLECULAR MODELING STUDIES OF PAMAM DENDRIMER PRODRUG POTENTIALLY ACTIVE IN TUBERCULOSIS

RENAN VINICIUS DE ARAÚJO(IC), DÉBORA FELICIANO SAVINO(IC), JEANINE GIAROLLA VARGAS

Departament of Pharmacy, FCF/USP

Introduction and Objectives: Tuberculosis is a deadly disease which in 2015, 10.4 million of new cases were reported and 1.8 million of deaths were registered, putting it in the top of ten death causes in worldwide.Dendrimers are a type of polymeric molecules, characterized as hyperbranched molecules, with a core from which new layers and terminal groups arise. The objective of this work is the use of molecular modeling simulations to predict the dendrimer cleavage and disassembly against a non-specific esterase, butyrilcholinesterase. The dendrimer studied is a PAMAM dendrimer using succinic acid as spacing agent and Isoniazid as terminal functions. Material and Methods: The three-dimensional models in their neutral forms were built up in Spartan 14, version 1.1.8. Partial atomic charges calculations and the energy-minimization were carried out in the same software. Docking simulations were performed in the GOLD 4.1.2, considering the catalytic site of the butyrilcholinesterase. 30 conformers were generated for each compound. The analysis was made considering the distances between the hydroxyl of serine 198 and carbonyls susceptible to enzymatic nucleophilic attack (Pymol software). The carbonyl atom near the isoniazid was numbered as C1 and the carbonyl closer to the dendrimer, was named as C2. Results and Conclusions: Docking demonstrated that C1 presented values between 1.1 to 3.0 Å and C2 from 0.6 to 2.2 Å. C2 seemed to be the most promising for suffering enzymatic action with a distances value of 0.6 Å, although cleavage on C1 doesn’t seem unlikely, presenting a low distance value of 1.1 Å. This data could indicate that the order in which the dendrimer is disassembled influence the cleavage activity of butyrilcholinesterase. This change in activity could possibly lead to different patterns of disassembly.Financing: FAPESP; CNPq


FCF133-2018HIGHER HYDROSOLUBILITY OF ROSUVASTATIN PRODRUGS

BRUNA MACHADO ARAÚJO SANCHES (PG); ELIZABETH IGNE FERREIRA

LAPEN, Pharmacy Department, FCF-USP

Introduction and Objectives: Developing process of innovative drugs faces many challenges, one of the most common is related to low aqueous solubility. Solubility is an essential property in drugs since it is a critical parameter, which directly influences bioavailability, affecting ADME properties. The prodrug approach is a molecular modification process that turns out to be a great strategy to modify physical-chemical properties, such as low aqueous solubility, without compromising pharmacological action of the parent compound. Rosuvastatin is a drug used in hypercholesterolemia and dyslipidemia treatment, being very important on prevention of cardiovascular diseases. However, it presents low solubility (0,0886 mg/mL) and oral bioavailability of only 20%. In view of the exposed and considering the importance of the prodrug approach on solving solubility issues, two rosuvastatin prodrugs were synthesized, using phosphate and 2,2-bis(hydroxymethylpropionic) acid (BisMPA) as carriers. Solubility studies and thermal analysis, beyond permeability assays, will confirm the achievement of the proposed objective. Material and Methods: The rosuvastatin ester prodrug synthesis have three steps. First, is necessary to protect the free hydroxyls on BisMPA in order to avoid cross reactions. Then, formation of the anhydride is better in order to achieve higher reactivity. So, the esterification can be done in rosuvastatin which can be mono or disubstituted. The rosuvastatin phosphate ester prodrug is synthesized by one-pot reaction using tetrabutylammonium hydrogen phosphate, as a phosphorylation reagent, and trichloroacetonitrile. Results and Conclusions: All compounds were characterized by NMR 1H and 13C. All the structures were confirmed. Financing: CNPq

FCF134-2018ANTIMALARIAL DRUG CANDIDATE: DESIGN AND SYNTHESIS OF PRIMAQUINE DENDRIMER

LUIS MIGUEL ZARAVIA ARGOMEDDD (D), ELIZABETH IGNE FERREIRA

Department of Pharmacy, Faculty of Pharmaceutical Sciences, University of Sao Paulo

Introduction and Objectives: Malaria has been a huge challenge for poor population in the world. About 216 million cases of this disease were registered in 2016. Although 2.7 billion US$ had been available for investment in this parasitosis, there are no drug completely efficient, especially because of the high level of resistant Plasmodium. Besides this, there is only one drug, primaquine, which has been active against the hepatic phase of this disease. However, primaquine presents some problems as short half-life and toxicity. Taking into account that dendrimers can be useful as drug carriers to solve many problems, including increase of drug half-life and decrease in toxicity, our objective was to synthesize dendrimers of primaquine, using malic acid as spacer group and glycerol as core. With the purpose of achieving selectivity to the liver, galactose, which can interact with this carbohydrate receptor in the hepatocyte membrane, was planned to be further linked to this dendrimer. Material and Methods: Malic acid linked to primaquine is the dendron and 1,3-dibromo-2-propanol was the core to make a selective and cleaner substitution to obtain compound, which is a first-generation dendrimer with one free side that can be further functionalized. Results and Conclusions: The compound obtained was analyzed by RMN 1H and 13C in Bruker Advance 300 MHz, and EM, in MICROTOF Bruker Daltonics. The synthesis of primaquine disubstituted dendrimers was achieved. In further experiments one more primaquine can be introduced. For targeted hepatic primaquine dendrimers, the primaquine disubstituted dendrimer will be linked to galactose, a directing group to be delivered selectively to hepatocytes. Financing: CAPES; CNPq


FCF135-2018INNOVATIVE APPROACH FOR THE TREATMENT OF OCULAR TUBERCULOSIS: RIFAMPICIN CATIONIC NANOEMULSION

MIRLA ANALI BAZAN HENOSTROZA (M), NADIA ARACI BOU-CHACRA

Department of Pharmacy, FCF/USP.

Introduction and Objectives: Ocular infection caused by Mycobacterium tuberculosis produce profound and irreversible loss of vision if it isn’t properly treated. The conventional treatment is the oral administration of the rifampin(R). Due to eye barriers, topical treatment requires high dose and repeated administrations to achieve a therapeutic effect on the eye. Considering these limitations, the present work aimed preparation, physicochemical characterization and microbiological evaluation of rifampicin nanoemulsion(RN) coated using chitosan(RN-Chit) and polymyxin B(RN-Poly) with potential application for topical treatment of ocular tuberculosis. Material and Methods: Based, on the solubility study of R, the RN was composed of oleic acid:polysorbate80(10:9) and ultrapure water. It was prepared by high-pressure homogenization method at 10,000Psi for 5cycles and coated using chitosan and polymyxin B. The coating process was conducted employing factorial experimental design. The particle size(z-ave), polydispersity index(PdI) and zeta potential(ZP) were measured using Nano ZS90. The MIC of preparations against Mycobacterium tuberculosis H37Rv strains was evaluated by colorimetric assay(MTT). Results and Conclusions: The RN presented z-ave, PdI, ZP and pH 137.3nm,0.19, -35.4mV and 5.26, respectively. The change in the ZP after the addition of chitosan was from -35.4 to +51.3mV. In the other case, the increased of polymyxin B changed the ZP from -35.4 to+5.5mV. Therefore, the methodological approach elucidated that the concentration of chitosan and polymyxin B significantly influenced the ZP of RN-Chit and RN-Poly. The MIC of RN, RN-Chit, RN-Poly and the standard solution of R was 0,125 µg/mL. This result demonstrated that the coating did not affect the antimicrobial efficacy of R. The present study allowed to obtain innovative preparations Financing: CAPES

FCF136-2018INHIBITION OF ANGIOTENSIN CONVERTING ENZYME ACTIVITY BY PHENOLIC COMPOUNDS FROM JABOTICABA (Plinia jaboticaba (Vell.) Berg) AFTER SIMULATED GASTROINTESTINAL DIGESTION

GABRIELA DE LIMA SANTIAGO (PG)*; PATRÍCIA ALESSANDRA BERSANETTI (PG)**; MARIA INÉS GENOVESE (PD)*

*Department of Food and Experimental Nutrition, University of São Paulo; *Department of Medicine, Federal University of São Paulo.

Introduction and Objectives: High blood pressure is responsible to cause about 13% of all deaths in the world, being the main risk factor for cardiovascular complications. Lifestyle changes and pharmacological therapies, such as the Angiotensin Converting Enzyme I (ACE) inhibitors, are among the treatments for this disorder. Researches have shown the benefits of phenolic compounds-rich diets in decrease the blood pressure. In this way, the aim of this work was to evaluate the inhibitory capacity of two different polyphenol-rich extracts obtained from in vitro digested jaboticaba, a Brazilian native fruit, on ACE activity in vitro. Material and Methods: For this purpose, the gastrointestinal system was simulated by adding digestive enzymes and controlling the pH of gastric and intestinal phase. Subsequently the digested content was centrifugated and its supernatant had their phenolic compounds purified by polyamide and octadecylsilane resins, that resulted in low and high tannin content extracts, respectively. The ACE assay was performed using a fluorimetric method. Results and Conclusions: Our results showed that the extract rich in tannins presented IC50 values almost five times lower than the extract with less content of these polyphenols, composed largely by flavonoids and phenolic acids. Moreover, a significant correlation was found among tannins and IC50 values, while phenolic acids and flavonoids didn’t show such correlation.Thus, jaboticaba tannins showed to be good inhibitors of in vitro ACE activity, but in vivo studies should be performed in order to know the real contribution of tannins on blood pressure control. Financing: CNPq; FAPESP.

AUTHOR INDEX/ÍNDICE DE AUTORES


AAGUIAR, M. J. FCF064-2018AGUIAR, P. M. FCF093-2018ALBUQUERQUE, C. N. FCF090-2018AMARAL, C. M. O. FCF115-2018, FCF120-2018ANTONIO, S. G. FCF025-2018APOLINÁRIO, A. C. FCF125-2018ARAÚJO, G. L. B. FCF015-2018, FCF024-2018,

FCF025-2018, FCF036-2018, FCF041-2018

ARAÚJO, N. T. FCF053-2018ARAÚJO, R. V. FCF132-2018ARGOMEDO, L. M. Z. FCF134-2018ARRUEBO, M. FCF030-2018ASSIS, S. R. FCF088-2018AURORA PRADO, M. S. FCF023-2018, FCF030-2018,

FCF035-2018AYALA, T. S. FCF080-2018

BBABY, A. R. FCF111-2018BALZI, A. P. C. C. FCF055-2018BARBOSA, E. G. FCF063-2018BARBOSA, L. R. S. FCF001-2018BAREA, B. FCF050-2018BARGIERI, D. FCF098-2018BARRIOS, J. O. FCF012-2018BARROS, S. B. M. FCF088-2018BATISTA, P. R. FCF068-2018BATTAGLIA, G. FCF102-2018BELEM, B. R. FCF028-2018BELLA, L. M. FCF080-2018BERSANETTI, P. A. FCF136-2018BEZZON, V. D. N. FCF015-2018, FCF106-2018,

FCF110-2018BIASOTO, H. FCF021-2018BOGSAN, C. S. B. FCF065-2018, FCF070-2018,

FCF075-2018BONEZI, V. FCF082-2018BONIFACE, P. K. FCF119-2018

BONVINI, A. FCF003-2018BORGES, D. O. FCF008-2018BORGES, F. M. FCF008-2018BOU-CHACRA, N. A. FCF024-2018, FCF094-2018,

FCF135-2018BRAGA, D. FCF026-2018BRAGA, M. S. FCF087-2018, FCF115-2018,

FCF116-2018, FCF120-2018BRANDÃO, W. FCF123-2018BRAZ, C. A. FCF063-2018, FCF069-2018BRESSAN, G. L. FCF060-2018BRITO, C. L. FCF042-2018BRUNO, V. FCF083-2018, FCF096-2018,

FCF118-2018BUENO, C. Z. FCF102-2018BURGER, M. FCF082-2018

CCAJAIBA, L. M. FCF050-2018CALVO, P. V. C. FCF029-2018CAMARGO, T. M. FCF078-2018CAMARGO, T. V. FCF055-2018CAMARINI, R. FCF083-2018, FCF096-2018,

FCF118-2018CAMPOS, G. Z. FCF114-2018CAMPOS, I. C. C. FCF092-2018, FCF107-2018CARRETERO, G. FCF044-2018, FCF112-2018CARVALHO, F. M. S. FCF024-2018, FCF036-2018CARVALHO, G. C. FCF087-2018, FCF120-2018CARVALHO, J. C. M. FCF056-2018, FCF086-2018CASAGRANDE, F. B. FCF075-2018CASTRO, I. FCF050-2018, FCF051-2018CASTRO, R. D. FCF001-2018CASTRO-ALVES, V. C. FCF129-2018CAVICHON, E. G. FCF066-2018, FCF099-2018CERIZE, N. N. P. FCF116-2018, FCF128-2018CHIARETO, J. F. FCF008-2018CHOW, V. S. S. FCF092-2018CONSOLINI, G. FCF002-2018COQUEIRO, A. FCF003-2018



CORDEIRO, E. W. F. FCF049-2018CORSI, G. C. FCF121-2018CORTEZ, M. FCF082-2018COSTA, I. M. FCF014-2018, FCF105-2018COSTA, L. M. FCF010-2018

DDARTORA, V. F. C. FCF131-2018DI MASCIO, P. FCF049-2018DIAS, G. F. FCF019-2018DIAS, M. V. B. FCF073-2018DOMINGUEZ, M. FCF098-2018DONADO-PESTANA, C. M. FCF007-2018DUARTE, N. J. FCF022-2018, FCF052-2018,

FCF054-2018, FCF055-2018DUCATI, L. C. FCF068-2018DURÃO, A. C. FCF123-2018DU-THUMM, L. FCF088-2018

EEFFER, B. FCF032-2018EPIPHANIO, S. FCF018-2018ESERIAN, J. K. FCF103-2018ESPÍRITO SANTO, É. FCF019-2018

FFABI, J. P. FCF017-2018, FCF109-2018FABRIS, A. L. FCF117-2018FAJARDO, F. A. G. FCF035-2018FANIZZI, J. G. FCF069-2018FARSKY, S. H. P. FCF117-2018FARSKY, S. FCF079-2018FEFERBAUM, R. FCF005-2018FEITOSA, I. FCF079-2018FEITOSA, V. A. FCF111-2018, FCF128-2018FERNANDES, A. P. S. FCF005-2018FERNANDES, T. B. FCF074-2018FERNÁNDEZ, J. A. FCF030-2018

FERRAZ, H. G. FCF025-2018, FCF028-2018, FCF041-2018, FCF106-2018, FCF110-2018, FCF126-2018

FERREIRA, E. I. FCF042-2018, FCF119-2018, FCF122-2018, FCF124-2018, FCF133-2018, FCF134-2018

FERREIRA, F. F. FCF041-2018FERREIRA, K. S. FCF080-2018FERREIRA, L. C. FCF079-2018, FCF082-2018FERREIRA, S. S. FCF075-2018FILGUEIRAS, L. R. FCF077-2018FORMAGIO, L. C. V. FCF115-2018FORSTER, C. FCF043-2018, FCF111-2018FORTES, A. C. FCF126-2018FRANCO, B. D. G. M. FCF114-2018FRANÇOSO, K. S. FCF078-2018FREITAS, P. D. C. FCF026-2018

GGABRIEL, F. C. FCF004-2018GARCIA, R. FCF083-2018GARZÓN, R. FCF047-2018GAZZINELLI, R. FCF098-2018GENOVESE, M. I. FCF007-2018, FCF009-2018,

FCF039-2018, FCF136-2018GHABDAN, N. FCF123-2018GHISLENI, D. D. M. FCF116-2018GIACOMOLLI, J. A. FCF008-2018GIMENEZ, A. M. FCF016-2018GIOIELLI, L. A. FCF084-2018GIUNTINI, E. B. FCF095-2018GLORIA, P. J. T. FCF113-2018GODOY, R. V. C. FCF034-2018GOMES, E. A. C. FCF019-2018, FCF056-2018GOMES, L. R. FCF095-2018GOMEZ, D. S. FCF022-2018, FCF052-2018,

FCF054-2018, FCF055-2018GONZAGA, R. V. FCF122-2018, FCF124-2018GRAYSON, S. FCF072-2018GUBITOSO, M. R. FCF024-2018



GUERRA-SHINOHARA, E. M. FCF127-2018GUIMARÃES, J. P. T. FCF077-2018GUMBREVICIUS, I. FCF095-2018

HHACKER, S. S. FCF065-2018HEIDOR, R. FCF109-2018HELUANY, C. FCF079-2018HENOSTROZA, M. A. B. FCF135-2018HERSZKOWICZ, P. S. D’O. FCF113-2018HIEDA, D. S. FCF088-2018HIRATA, R. D. C. FCF046-2018HO, D. C. FCF097-2018HOFFMANN, C. FCF095-2018, FCF113-2018,

FCF114-2018HOFFMANN-SARDA, F. A. FCF095-2018, FCF113-2018HÜBNER, A. A. FCF026-2018

IISHII, M. FCF019-2018, FCF056-2018ISSA, M. G. FCF028-2018, FCF106-2018,

FCF126-2018

JJANCAR, S. FCF077-2018JANUZZI, G. P. FCF080-2018JEANINE GIAROLLA VARGAS FCF072-2018, FCF132-2018JESUS, M. D. R. FCF046-2018

KKANEKO, T. M. FCF103-2018KATO, E. M. FCF026-2018KEDOR-HACKMANN, E. R. M. FCF035-2018KIKUCHI, I. S. FCF053-2018KISHIMA, E. FCF028-2018, FCF106-2018KITIZABOLO, A. C. O. FCF039-2018KNÖBL, T. FCF089-2018KOIAMA, J. R. FCF118-2018KOIAMA, J. FCF083-2018, FCF096-2018

KUNIYOSHI, T. M. FCF031-2018, FCF085-2018, FCF101-201, FCF104-2018

KUPA, L. V. K. FCF052-2018, FCF054-2018

LLACORTE, G. A. FCF114-2018LANDGRAF, M. FCF114-2018, FCF108-2018LANNES, S. C. S. FCF047-2018, FCF092-2018,

FCF107-2018LA-SCALEA, M. A. FCF042-2018LEÃO, C. C. FCF041-2018LEITE, H. O. A. FCF071-2018LEITE, L. R. FCF064-2018LEITE, S. N. FCF100-2018LÉO, P. FCF111-2018, FCF116-2018,

FCF128-2018LEON, R. M. FCF110-2018LIMA, D. FCF082-2018LIMA, E. I. A. FCF120-2018LIMA, E. M. FCF023-2018LIMA, E. S. FCF037-2018LIMA, F. S. FCF113-2018LIMA, G. M. FCF032-2018LIMA, T. M. FCF093-2018LINDOSO, J. A. L. FCF063-2018LIRA, R. S. FCF031-2018LOMBARDO, M. FCF103-2018LOPES, L. B. FCF131-2018LOPES, R. G. FCF017-2018LOTIERZO, M. C. G. FCF001-2018LOUREIRO, A. P. M. FCF049-2018LOURENÇO, F. R. FCF011-2018, FCF027-2018,

FCF026-2018, FCF048-2018, FCF087-2018, FCF103-2018

LOUSADA, M. E. G. FCF019-2018, FCF056-2018

MMAFIOLETTI, M. C. FCF006-2018MALBOUISSON, L. M. FCF055-2018MALHEIROS, B. FCF001-2018



MANCINI FILHO, J. FCF006-2018, FCF121-2018MANFRINATO, C. V. FCF065-2018MARCONDES, A. L. K. FCF020-2018MARCOURAKIS, T. FCF118-2018, FCF083-2018,

FCF096-2018, FCF123-2018MARIA-ENGLER, S. S. FCF049-2018, FCF088-2018MARQUES, R. F. FCF016-2018, FCF078-2018MARTINS, J. O. FCF076-2018, FCF077-2018,

FCF080-2018 MARTINS, R. FCF033-2018MASSARETTO, I. L. FCF008-2018MATSUO, M. M. FCF065-2018MEBIAME, G. E. FCF069-2018MEDEIROS, M. H. G. FCF049-2018MELLO, B. V. FCF088-2018MELO, D. O. FCF004-2018MELO, I. L. P. FCF006-2018, FCF121-2018MELO, K. J. C. FCF106-2018MEM, A. FCF108-2018MENDES, S. J. FCF100-2018MENEZES, E. W. FCF095-2018MESSIANO, C. G. FCF054-2018MEZA, S. L. R. FCF008-2018MITIKO. M. FCF075-2018MOLINO, C. G. R. C. FCF004-2018MONTEMOR, A. F. FCF128-2018MORAES, P. FCF026-2018MORALES, G. S. FCF053-2018MOREIRA, L. N. FCF005-2018MORENO, F. S. FCF109-2018MORO, L. FCF008-2018MOURA, D. C. FCF014-2018MOURA, M. H. C. FCF009-2018, FCF039-2018MOURA, W. A. FCF090-2018

NNAKAYA, H. FCF082-2018NASCIMENTO, C. FCF033-2018NASCIMENTO, J. R. O. FCF129-2018NASCIMENTO, L. A. FCF122-2018, FCF124-2018

NASCIMENTO, L. C. F. FCF035-2018NASCIMENTO, P. C. FCF127-2018NAZARETH JR., E. M. FCF036-2018NEUGEBAUER, V. R. FCF096-2018, FCF118-2018NEUGEBAUER, V. FCF083-2018NEVES, C. T. C. FCF005-2018, FCF033-2018,

FCF034-2018NOGUEIRA, M. S. FCF051-2018NOVAIS, J. T. FCF016-2018, FCF078-2018NUNES, A. V. FCF079-2018NUNES, F. P. B. FCF075-2018

OOKAMOTO, R. T. FCF087-2018OLIVEIRA, A. M. FCF043-2018, FCF116-2018OLIVEIRA, C. A. FCF043-2018, FCF044-2018,

FCF111-2018, FCF128-2018, OLIVEIRA, D. M. S. FCF090-2018OLIVEIRA, D. M. FCF009-2018, FCF039-2018OLIVEIRA, J. FCF049-2018OLIVEIRA, R. E. FCF113-2018OLIVEIRA, R. P. S. FCF031-2018, FCF085-2018,

FCF089-2018, FCF091-2018, FCF101-2018, FCF104-2018

OLIVEIRA, R. P. S. FCF130-2018ONG, T. P. FCF058-2018

PPACHECO, L. T. FCF083-2018PALMA, M. S. A. FCF002-2018, FCF020-2018,

FCF029-2018, FCF038-2018, FCF045-2018, FCF059-2018, FCF060-2018, FCF061-2018, FCF062-2018, FCF064-2018, FCF068-2018

PARISE FILHO, R. FCF012-2018, FCF057-2018, FCF067-2018, FCF073-2018, FCF074-2018

PASCOAL, G. B. FCF008-2018PASSOS, J. S. FCF131-2018



PAULA-SILVA, M. FCF117-2018PEREIRA, E. F. FCF081-2018PEREIRA, G. J. V. FCF067-2018PEREIRA, L. FCF082-2018PERON, J. P. FCF123-2018PESSOA JUNIOR, A. FCF014-2018, FCF099-2018,

FCF102-2018, FCF125-2018PESSOA, C. A. FCF002-2018, FCF059-2018PESSOA, É. V. M. FCF007-2018PIAZENTIN, A. C. M. FCF091-2018PICOSSI, C. C. FCF035-2018PINTO, C. F. F. FCF048-2018PINTO, J. T. F. FCF048-2018PINTO, T. J. A. FCF087-2018, FCF115-2018,

FCF120-2018PINTO, U. M. FCF114-2018PRADO, F. K. FCF023-2018, FCF030-2018PRADO, S. B. R. FCF109-2018PRESTES, A. FCF075-2018PRIETO, D. C. FCF072-2018PURGATTO, E. FCF008-2018, FCF037-2018

QQUIRINO, T. C. FCF018-2018

RRACT, J. N. R. FCF084-2018RAIZEL, R. FCF003-2018RANGEL YAGUI, C. O. FCF043-2018, FCF044-2018,

FCF066-2018, FCF099-2018, FCF102-2018, FCF111-2018, FCF112-2018, FCF125-2018, FCF128-2018

RECHE, L. L. FCF045-2018REPIN, I. A. FCF025-2018, FCF041-2018RIBEIRO, E. FCF004-2018RIBEIRO, G. M. FCF073-2018RIBEIRO, J. A. FCF073-2018RIOS, R. V. FCF047-2018ROCHA, B. M. FCF044-2018

ROCHA, G. H. O. FCF117-2018RODRIGUES, Á. G. FCF090-2018RODRIGUES, G. F. FCF001-2018RODRIGUES, L. FCF007-2018ROGERO, M. FCF003-2018ROSELL, C. M. FCF047-2018

SSABO, S. S. FCF031-2018, FCF085-2018,

FCF089-2018SAITO, D. H. N. FCF062-2018SANCHES, B. M. A. FCF133-2018SANSÓN, M. D. S. FCF084-2018SANT’ANNA, C. L. FCF086-2018SANTAMARÍA, J. FCF030-2018SANTIAGO, G. L. FCF136-2018SANTORO, M. I. R. M. FCF035-2018SANTOS JR., E. N. FCF038-2018SANTOS, D. O. FCF061-2018SANTOS, J. H. P. M. FCF044-2018, FCF066-2018,

FCF099-2018, FCF112-2018SANTOS, J. L. FCF028-2018, FCF126-2018SANTOS, M. R. FCF049-2018SANTOS, N. C. L. FCF004-2018SANTOS, R. M. C. FCF116-2018SANTOS, S. R. C. J. FCF022-2018, FCF052-2018,

FCF054-2018, FCF055-2018SANTOS, V. G. FCF085-2018SANTOS, W. C. FCF127-2018SATTLER, J. A. G. FCF006-2018, FCF121-2018SAVIANO, A. M. FCF027-2018SAVINO, D. F. FCF132-2018SCOLARO, B. FCF051-2018SEBASTIAN, V. FCF030-2018SEPAROVIC, L. FCF011-2018SERRA, C. H. R. FCF036-2018SILVA JR., E. M. FCF022-2018, FCF052-2018,

FCF054-2018SILVA JR., J. M. FCF022-2018, FCF052-2018,

FCF054-2018



SILVA, A. R. P. FCF066-2018SILVA, A. T. FCF125-2018SILVA, C. FCF105-2018SILVA, G. C. FCF049-2018SILVA, G. K. FCF090-2018SILVA, L. B. A. R. FCF058-2018SILVA, L. C. M. FCF086-2018SILVA, N. A. T. F. FCF057-2018SILVA, R. A. M. FCF004-2018SILVA, R. R. O. FCF020-2018, FCF029-2018,

FCF038-2018, FCF045-2018, FCF059-2018, FCF061-2018, FCF068-2018

SILVA, T. M. S. FCF130-2018SILVA, V. C. FCF058-2018SILVEIRA, E. L. V. FCF117-2018SILVEIRA, E. FCF079-2018, FCF082-2018,

FCF098-2018SILVEIRA, T. FERREIRA FCF050-2018SIMIONATO, M. R. L. FCF113-2018SINGH, A. K. FCF010-2018, FCF040-2018,

FCF056-2018, FCF071-2018SOARES, I. S. FCF016-2018, FCF078-2018,

FCF098-2018SOUZA, A. D. F. FCF073-2018SOUZA, C. A. FCF070-2018SOUZA, G. M. FCF013-2018, FCF014-2018,

FCF021-2018, FCF032-2018, FCF097-2018, FCF105-2018

SOUZA, I. G. FCF087-2018SOUZA, L. F. FCF090-2018SOUZA, V. K. FCF022-2018SPARVOLI, L. G. FCF034-2018STORPIRTIS, S. FCF093-2018, FCF100-2018

TTAMAROSSI, R. I. FCF109-2018TAVARES, L. C. FCF063-2018, FCF069-2018TAVARES, M. F. M. FCF030-2018TAVARES, M. F. M. FCF035-2018TESSARO, F. H. G. FCF080-2018TIRAPEGUI, J. FCF003-2018TOBARUELA, E. C. FCF008-2018, FCF037-2018TODOROV, S. D. FCF065-2018TROSSINI, G. H. G. FCF073-2018, FCF081-2018TSUJITA, M. FCF075-2018

UUONO, M. T. FCF065-2018

VVALENCIA, A.O. FCF013-2018VASCONCELOS, L. P. FCF004-2018VENTURA, S. P. M. FCF099-2018FCF112-2018VIEIRA, E. S. FCF084-2018VIEIRA, T. M. FCF051-2018VIERA, V. B. FCF101-2018, FCF104-2018VILLENEUVE, P. FCF050-2018

WWAINBERG, S. K. FCF004-2018WU, J. FCF088-2018

YYATACO-LAZARO, L. M. FCF030-2018YOSHIDA, B. FCF075-2018YOSHIME, L. T. FCF006-2018, FCF121-2018YUKUYAMA, M. N. FCF094-2018

Revista Brasileira de Ciências FarmacêuticasBrazilian Journal of Pharmaceutical Sciences

Scientific EditorElizabeth Igne Ferreira

Associated EditorsProf. Carlota Oliveira Rangel YaguiProf. Cristina Helena dos Reis Serra Prof. Elfriede Marianne Bacchi Prof. Eliezer Jesus de Lacerda Barreiro Prof. Felix Carvalho Prof. Flávio Finardi Filho

Editorial Advisory Board Adriano Delfini Andricopulo Instituto de Física de São Carlos/Universidade de São Paulo Alan Douglas Kinghorn College of Pharmacy/The Ohio State University, USA Anton Joseph Hopfinger College of Pharmacy/University of Illinois at Chicago, USA Antonio Flávio Mídio Faculdade de Ciências Farmacêuticas/Universidade de São Paulo (aposentado) Antonio Monge Vega Facultad de Farmacia/Universidad de Navarra, España Antônio Salatino Instituto de Biociências/Universidade de São Paulo Attilio Converti Universitá Degli Studi di Genova, Italy Camille-Georges Wermuth Prestwick Chemical, France Carol Hollingworth Collins Instituto de Química/Universidade Estadual de Campinas Dulcinéia Saes Parra Abdalla Faculdade de Ciências Farmacêuticas/Universidade de São Paulo Elfrides Eva Scherman Schapoval Faculdade de Farmácia/Universidde Federal do Rio Grande do Sul Eliezer de Jesus Lacerda Barreiro Faculdade de Farmácia/Universidade Federal do Rio de Janeiro Eloir Paulo Schenkel Centro de Ciências da Saúde/Universidade Federal de Santa Catarina Francisco José Baptista Veiga Faculdade de Farmácia/Universidade de Coimbra, Portugal Franco Maria Lajolo Faculdade de Ciências Farmacêuticas/Universidade de São Paulo Isac de Almeida Medeiros Centro de Ciências da Saúde/Universidade Federal da Paraíba Isarita Martins Faculdade de Farmácia/Universidade Federal de Alfenas Ivan da Rocha Pitta Centro de Ciências Biológicas/Universidade Federal de Pernambuco João Luis Callegari Lopes Faculdade de Ciências Farmacêuticas/Universidade de São Paulo, Ribeirão Preto Jorge Mancini Filho Faculdade de Ciências Farmacêuticas/Universidade de São Paulo Lolita Margareta Tsanaclis Cansford Lab. Limited (UK) e Laboratórios Chromatox Ltda. (São Paulo) Marcos Alcocer School of Biosciences/University of Nottingham, United Kingdom Maria Helena Gil Faculdade de Ciências e Tecnologia/Universidade de Coimbra,Portugal Maria Inês Rocha Miritello Santoro Faculdade de Ciências Farmacêuticas/Universidade de São Paulo (aposentada) Maria José Soares Mendes Gianini Faculdade de Ciências Farmacêuticas/UNESP“Julio de Mesquita Filho”, Araraquara Maria Nella Gai Facultad de Ciencias Quimicas y Farmacéuticas/Universidad de Chile, Chile Marilene De Vuono Camargo Penteado Faculdade de Ciências Farmacêuticas/Universidade de São Paulo Massayoshi Yoshida Instituto de Química/Universidade de São Paulo (aposentado) Michele Vitolo Faculdade de Ciências Farmacêuticas/Universidade de São Paulo Mumtaz Iscan Faculty of Pharmacy/Ankara University, Turkey Paulo César Stringheta Centro de Ciências Biológicas e da Saúde/Universidade Federal de Viçosa Richard A. Glennon School of Pharmacy/Virginia Commonwealth University, USA Robert Dennis Tanner Vanderbilt University, USA (Emeritus professor) Roy Edward Bruns Instituto de Química/Universidade Estadual de Campinas Rui Curi Instituto de Ciências Biomédicas/Universidade de São Paulo Seizi Oga Faculdade de Ciências Farmacêuticas/Universidade de São Paulo (aposentado) Sílvia Berlanga de Moraes Barros Faculdade de Ciências Farmacêuticas/Universidade de São Paulo (aposentado) Silvia Storpirtis Faculdade de Ciências Farmacêuticas/Universidade de São Paulo Suely Vilela Faculdade de Ciências Farmacêuticas/Universidade de São Paulo, Ribeirão Preto Tasso Moraes e Santos Faculdade de Farmácia/Universidade Federal de Minas Gerais Tudor Oprea School of Medicine/University of New Mexico, USA Victor Manuel Cardoso Figueiredo Balcão Universidade de Sorocaba (UNISO) Yves Le Loir Institut National de la Recherche Agronomique, France

Prof. Lídia Moreira Lima Prof. Luis Antonio Salazar Navarrete Prof. Luiz Antonio Gioielli Prof. Priscila Gava Mazzola Prof. Sílvia Berlanga de Moraes Barros Prof. Sílvia Storpirtis Prof. Vladi O. Consiglieri

Technical and Administrative SupportDivisão de Biblioteca e Documentação do Conjunto das Químicas da USP

2

SCOPE AND POLICY

The Brazilian Journal of Pharmaceutical Sciences (BJPS) is a peer-reviewed electronic journal published quarterly by the School of Pharmaceutical Sciences of the University of São Paulo.

The purpose of the Brazilian Journal of Pharmaceutical Sciences is to publish manuscripts that significantly contribute to knowledge in all areas of Pharmaceutical Sciences, including Medicines and Drugs, Pharmaceutical and Health Care, Food and Experimental Nutrition, Clinical Chemistry, Toxicology, Medicinal Chemistry, Pharmaceutical Technology, Biotechnology among others.

The following papers will not be accepted for publication: • Studies on human subjects not approved by an accredited

Ethics Committee or without written informed consent from the subject or legal guardian.

• Studies on animals not approved by an accredited Ethics and Animal Care Committee.

• Manuscripts describing plant extract activity that do not identify quali and quantitative markers of the extract.

PREPARATION OF THE MANUSCRIPTS

Manuscripts that do not agree to the Instructions will be refused prior to peer review.

Manuscripts must be submitted in English. Submission of a manuscript to BJPS implies that the data have not

been published previously and will not be submitted for publication elsewhere while the manuscript is under review.

Co-authors should be individuals who have contributed substantially to the content of the paper.

Manuscripts in accordance to the “Preparing your manuscript section” will be submitted for peer review to at least two independent, anonymous referees indicated by the Associated Editors. Based on peer review, the Associate Editors will suggest manuscript acceptance or not to the Editor-in-Chief, who is responsible for the final decision.

In the case revision is suggested, the authors are asked to resubmit the manuscript incorporating the suggestions and recommendations of the referees within 15 calendar days. If the revised version is not received within the time specified from the date of notice, the manuscript process will be canceled. All revisions must be accompanied with a letter detailing the changes made to the original document and answering all the reviewer comments, on a point-by-point basis. All alterations must be identified in the revised manuscript.

Manuscripts must have their copyright assigned to the BJPS before submitting to the Journal.

The dates of receipt and acceptance will be published for each article. Authors are expected to return reviewed manuscripts to the Journal within 15 calendar days, and to return galley proofs of accepted manuscripts within 72 hours. The total number of “late” days will be added to the submission date at the time of publication.

Authors are required to suggest 4 potential reviewers with information of institutional and e-mail address. At least 2 of the potential reviewers suggested should be from a different country to the corresponding authors. The Editors reserve the right to indicate these or other reviewers for manuscript evaluation.

MANUSCRIPT CATEGORIES

The authors should state in the cover letter that the manuscript is intended to be Full-length Original Paper, Short Communication, Review Article, Mini-review article, Concepts and Comments and Book Reviews.

The Journal will also publish Thematic or Congress Abstracts Supplements under invitation by the Editors or previous approval of the Editorial Board.

BJPS will publish the following type of articles:

Full-length Original PaperEach manuscript should clearly state its objective or hypothesis;

the experimental design and methods used; the essential features of any interventions; the main outcome measures; the main results of the study; and a discussion placing the results in the context of published literature.

The manuscript should contain:• abstract of no more than 250 words• no more than 6 key words• a running title to be used as a page heading, which should not

exceed 60 letters and spaces• manuscript main body divided into separate sections

(Introduction, Material and Methods, Results and Discussion).• no more than 40 references (without exceptions)• Supplementary data can be submitted as Suppmentary

information session.

Short CommunicationA short communication is a report on a single subject, which

should be concise but definitive. The scope of this section is intended to be wide and to encompass methodology and experimental data on subjects of interest to the readers of the Journal.

The manuscript should contain:• abstract of no more than 250 words• no more than 6 key words• a running title to be used as a page heading, which should not

exceed 60 letters and spaces• manuscript main body divided into separate sections

(Introduction, Material and Methods, Results and Discussion), without a separate section for conclusions

• no more than 20 references (without exceptions)• no more than three illustrations (figures and/or tables)

Review ArticleA review article should provide a synthetic and critical analysis of

a relevant area and should not be merely a chronological description of the literature. A review article by investigators who have made substantial contributions to a specific area of Pharmaceutical Sciences will be published by invitation of the Editors. However, an outline of a review article may be submitted to the Editors without prior consultation. If it is judged appropriate for the Journal, the author(s) will be invited to prepare the article for peer review. The manuscript should contain:

• abstract of no more than 250 words• no more than 6 key words• a running title to be used as a page heading, which should not

exceed 60 letters and spaces

3

• manuscript main body divided into sections with appropriate titles and subtitles

• no more than 90 references (without exceptions)

Mini-review ArticleA mini-review is focused on a restricted part of a subject normally

covered in a review article. The structure of the mini-review follows the same rules as the review.

Concepts and CommentsThe Concepts and Comments section provides a platform for

readers to present ideas, theories and views.The manuscript should contain:• abstract of no more than 250 words• no more than 6 key words• a running title to be used as a page heading, which should not

exceed 60 letters and spaces• manuscript main body divided into sections with appropriate

titles and subtitles• no more than 40 references (without exceptions)

Book ReviewsWritten by experts indicated by the Editors or written by the

authors.

PREPARING YOUR MANUSCRIPT

Cover LetterIt is important that you include a cover letter with your manuscript.

Take the time to consider why this manuscript is suitable for publication in the Brazilian Journal of Pharmaceutical Sciences. Why will your paper inspire other members of your field, and how will it drive research forward? Please explain this in your cover letter.

The cover letter should also contain the following information:• Title of article.• Name(s) of all author(s).• Information of Corresponding Author (name, full address,

telephone number and e-mail).

Authorship requirementsOnly people who directly contributed to the intellectual content

of the paper should be listed as authors. All manuscripts must be, submitted, only, by electronic way. The confirmation of submission is sent by email for all the authors, for their agreement.

Authors should meet all of the following criteria, thereby taking public responsibility for the content of the paper:

• Conceived, planned and carried out the experiments presented in the manuscript or interpreted the data, or both.

• Wrote the paper, or reviewed successive versions.• Approved the final version.• Holding positions of administrative leadership, contributing

patients, and collecting and assembling data, however important to the research, are not by themselves criteria for authorship. Any person who has made substantial, direct contribution to the work but cannot be considered an author should be cited in the Acknowledgment section, with permission and a description of his/her specific contribution to the research.

Text format• The text of a manuscript can only be accepted as a Microsoft

Word file created with MS Word as a “doc”, “docx” or “rtf” document.

• Manuscripts should be sent in 30-36 lines, 1,5 spaced,• Each page should contain the page number in the upper right-

hand corner starting with the title page as page 1.• Report all measurements in Système International, SI

(http://physics.nist.gov/cuu/Units) and standard units where applicable

• Names of plants, animals and chemicals should be mentioned according to International Rules available.

• Names of drugs can follow the International rules (DCI) or current Brazilian rules (DCB)

• Trademarks may be mentioned only once in the text (between parenthesis and initial in capital letter)

• Do not use abbreviations in the title and limit their use in the abstract and text.

• The length of the manuscript and the number of tables and figures must be kept to a minimum.

• Ensure that all references are cited in the text.• Generic names must be used for all drugs. Instruments may be

referred to by proprietary name; the name and country of the manufacturer should be given in parenthesis.

ORGANIZATION OF THE MANUSCRIPT

Most articles published in BJPS will be organized into the following sections:• Title, Authors, Abstract, Key words, Running Title, Author for Correspondence and email address• Introduction• Material and Methods• Results and Discussion• Acknowledgments• References• Tables with a descriptive title and footnote legends• Figures with a descriptive title, descriptive legends and uniformity in format

Continuous page numbers are required for all pages including figures. There are no specific length restrictions for the overall manuscript or individual sections. However, we urge authors to present and discuss their findings concisely. We recognize that some articles will not be best presented in our research article format. If you have a manuscript that would benefit from a different format, please contact the editors to discuss this further.

Title PageTitle - The title should be as short and informative as possible,

should not contain non-standard acronyms or abbreviations, and should not exceed two printed lines.

Examples:Freeze-drying of ampicillin solid lipid nanoparticles using mannitol

as cryoprotectantA fully validated microbiological assay for daptomycin injection

and comparison to HPLC methodPharmacokinetics, safety and tolerability of L-3-n-butylphthalide

http://physics.nist.gov/cuu/Units/

4

tablet after single and multiple oral administrations in healthy Chinese volunteers.

Authors and AffiliationsFull name (matched with superscript numbers identifying

affiliation). Institution(s) (Department, Faculty, University, City, State, Country) of each author (in English).

Examples: Hongmei Xia1 * , Yongfeng Cheng2 , Yinxiang Xu3 , Zhiqing

Cheng1

1College of Pharmacy, Anhui University of Chinese Medicine, Hefei, People’s Republic of China.

2School of Life Science, University of Science and Technology of China, Hefei, People’s Republic of China.

3Zhaoke (Hefei) Pharmaceutical Co. Ltd., Hefei, People’s Republic of China.

AbstractSince abstracts are published separately by Information Services,

they should contain sufficient hard data to be appreciated by the reader. The abstract should not exceed 250 words and should be prepared in a single paragraph.

The abstract should briefly and clearly present the objective, experimental approach, new results as quantitative data if possible, and conclusions. It should mention the techniques used without going into methodological detail and mention the most important results.

Abbreviations should be kept to a minimum and should be defined in both the Abstract and text. Please do not include any reference citations in the abstract. If the use of a reference is unavoidable, the full citation should be given within the abstract.

Key WordsA list of key words or indexing terms (no more than 6) should be

included avoiding generic terms.

Running titleThis short title, to be used as a page heading, should not exceed

60 letters and spaces.

Corresponding authorOne of the authors should be designated as the corresponding

author. It is the corresponding author´s responsibility to ensure that the author list is accurate and complete. If the article has been submitted on behalf of a consortium, all consortium members and affiliations should be listed in the Acknowledgments section. Provide the name and email address of the author to whom correspondence should be sent identified with an asterisk.

Introduction The Introduction should put the focus of the manuscript into a

broader context and reflects the present state-of-art of the subject. This should state briefly and clearly the objectives of the investigation with reference to previous works. Extensive review of the literature should be avoided and substituted for references of recent review publications.

Material and MethodsThese should be described in sufficient detail that the work can be

reproduced. Well-established procedures and techniques require only a citation of the original source, except when they are substantially modified. Reports of experimental studies on humans and animals must certify that the research received prior approval by the appropriate institutional review Ethics Committee.

Results and DiscussionResults must be presented clearly and concisely and in logical order.

This section should provide results of all of experiments required to support the conclusions of the paper. When possible, use figures or tables to present data rather than text. Large datasets, including raw data, should be submitted as supplementary files; these are published online linked to the article. Discussion should interpret the results and assess their significance in relation to existing knowledge. Speculation not warranted by actual data should be avoided. The Discussion should spell out the major conclusions and interpretations of the work including some explanation of the significance of these conclusions.

Acknowledgments When appropriate, briefly acknowledge technical assistance,

advice and contributions from colleagues. People who contributed to the work but do not fit the criteria for authors should be listed in the Acknowledgments section, along with their contributions. Donations of animals, cells, or reagents should also be acknowledged. You must also ensure that anyone named in the Acknowledgments agrees to being so named. Financial support for the research and fellowships should be acknowledged in this section (agency and grant number).

FiguresFigures must be submitted in high-resolution version (600 dpi).

Preparing figure files for submissionBJPS encourages authors to use figures where this will increase

the clarity of an article. The use of color figures in articles is free of charge. The following guidelines must be observed when preparing figures. Failure to do so is likely to delay acceptance and publication of the article.

• Each figure of a manuscript should be submitted as a single file.

• Figures should be numbered in the order they are first mentioned in the text, and uploaded in this order.

• Figure titles and legends should be provided in the main manuscript as a List of Figures, not in the graphic file.

• The aim of the figure legend should be to describe the key messages of the figure, but the figure should also be discussed in the text.

• An enlarged version of the figure and its full legend will often be viewed in a separate window online, and it should be possible for a reader to understand the figure without moving back and forth between this window and the relevant parts of the text.

• The legend itself should be succinct, while still explaining all symbols and abbreviations. Avoid lengthy descriptions of methods. Statistical information should be given as well as the statistical tests used.

• Arrows or letters should be used in the figure and explained in the legend to identify important structures.

5

• Figures with multiple panels should use capital letters A, B, C, etc. to identify the panels.

• Each figure should be closely cropped to minimize the amount of white space surrounding the illustration. Cropping figures improves accuracy when placing the figure in combination with other elements, when the accepted manuscript is prepared for publication.

• Individual figure files should not exceed 5 MB. If a suitable format is chosen, this file size is adequate for extremely high quality figures.

Please note that it is the responsibility of the author(s) to obtain permission from the copyright holder to reproduce figures (or tables) that have previously been published elsewhere. In order for all figures to be open-access, authors must have permission from the rights holder if they wish to include images that have been published elsewhere in non-open-access journals. Permission should be indicated in the figure legend, and the original source included in the reference list.

Supported file typeThe following file format can be accepted: TIFF (suitable for

images) or JPEG with 600 dpi, and Word file for the manuscript.

Tables• Tables must be submitted in Word (.doc) or Excel (.xls), not

as an image.• Tables must be numbered consecutively with Roman numerals

in the text.• Tables must have a concise and descriptive title.• All explanatory information should be given in a footnote

below the table. Footnotes should be used to explain abbreviations and provide statistical information, including statistical tests used.

• All abbreviations must be defined in this footnote, even if they are explained in the text.

• Tables must be understandable without referring to the text.• Tables occupying more than one printed page should be

avoided, if possible.• Vertical and diagonal lines should not be used in tables; instead,

indentation and vertical or horizontal space should be used to group data.

ReferencesReferences should be prepared and listed according to Vancouver

standard reference style. Entries should be arranged in alphabetical order by author at the end of the paper. All authors’ names should be given. Accuracy and completeness of reference data is the responsibility of the authors.

Only published references should be included in the reference list. Meeting abstracts, conference talks, or papers that have been submitted but not yet accepted should not be cited. Limited citation of unpublished work should be included in the body of the text only. All personal communications should be supported by a letter from the relevant authors.

References should be cited in the text by the authors’ names, with only the first letter in capital letter followed by the year of publication. For more than three authors, the first has to be cited followed by the expression et al. (in italic). Small letters close to the year must differentiate references of the same authors and year of publication.

Examples: (Fujisawa, Atsumi, Kadoma, 1989) (Aviral et al., 2009) (Dodu, Rotari, Vazques, 2012) (Liu et al., 2011a) (Liu et al., 2011b)

Please use the following style for the reference list:

Published Papers. First 6 authors followed by et al., Title, Journal (abbreviation in italic), Year, Volume, Complete Pages.

Abe T, Fukushima N, Brune K, Boehm C, Sato N, Matsubayashi H, et al. Genome‐Wide allelotypes of familial pancreatic adenocarcinomas and familial and sporadic intraductal papillary muninous neoplasms. Clin Cancer Res. 2007;13(20):6019‐25.

Ali A, Iqbal F, Taj A, Iqbal Z, Amin MJ, Iqbal QZ. Prevalence of microvascular complications in newly diagnosed patients with Type 2 diabetes. Pak J Med Sci. 2013,29(4): 899-902.

Calvo A, Gimenez MJ. Ex Vivo Serum Activity (Killing Rates) After Gemifloxacin 320 mg Versus Trovafloxacin 200 mg Single Doses Against Ciprofloxacin-Susceptible and -Resistant Streptococcus pneumoniae. Int J Antimicr Ag. 2007;20:144-6.

Lammers AE, Hislop AA, Flynn Y, Haworth SG. The 6-minute walk test: normal values for children of 4-11 years of age. Arch Dis Child. 2008;93:464-468.

Zhang Q, Malik P, Pandey D, Gupta S, Jagnandan D, Belin de CE, et al. Paradoxical activation of endothelial nitric oxide synthase by NADPH oxidase. Arterioscler Thromb Vasc Biol. 2008;28:1627-1633.

Article accepted for publication but not yet published. First 6 authors followed by et al., Title, Journal (abbreviation in italic), Year of expected publication, (in press) at the end of the citation.

Janiszewski M, Lopes LR, Carmo AO, Pedro MA, Brandes RP, Santos CXC, et al. Regulation of NAD(P)H oxidase by associated protein disulfide isomerase in vascular smooth muscle cells. J Biol Chem. 2005 (in press).

Internet Communication. Ensure that URLs are active and available. Provide DOI, if available.

Brasil. Ministério da Saúde, Secretaria de Vigilância em Saúde. Leishmaniose visceral grave: normas e condutas [Internet]. Brasília (DF): Ministério da Saúde, 2006. [citado 2008 Jan 7]. 60 p. (Série A. Normas e Manuais Técnicos). Disponível em: http://dtr2001.saude.gov.br/editora/produtos/livros/pdf/06_0072_M.pdf

CAPES Statistics. [cited 2006 Mar 16]. Available from: http://www.capes.gov.br/capes/portal.

Developmental toxicology. [cited 2015 Apr 10]. Available from: http://www.devtox.org/nomenclature/organ.php.

http://dtr2001.saude.gov.br/editora/produtos/livros/pdf/06_0072_M.pdf

http://dtr2001.saude.gov.br/editora/produtos/livros/pdf/06_0072_M.pdf

6

Book, Whole. Authors, Book title, Edition, City, Publisher, Year. Hewitt W. Microbiological assay for pharmaceutical analysis:

a rational approach. Boca Raton: CRC Press; 2003.

Jenkins PF. Making sense of the chest x-ray: a hands-on guide. New York: Oxford University Press; 2005. 194 p.

Milech A, et al., Oliveira JEP, Vencio S, organizadores. Diretrizes da Sociedade Brasileira de Diabetes. São Paulo: A.C. Farmacêutica;2016.

Book, Chapter. Authors, Chapter Title, Editors, Book title, Edition, City, Publisher, Year, Pages of citation.

Beizer JL, Timiras ML. Pharmacology and drug management in the elderly. In: Timiras PS, editor. Physiological basis of aging and geriatrics. 2nd ed. Boca Raton: CRC Press; 1994. p. 279-84.

Rojko JL, Hardy WD Jr. Feline leukemia virus and other retroviruses. In: Sherding RG, editor. The cat: diseases and clinical management. New York: Churchill Livingstone; 1989. p. 229-332.

Report World Health Organization. WHO. Department of Mental

Health and Substance Abuse. Mental health atlas 2005. Geneva: World Health Organization; 2005. 409 p.

World Health Organization. WHO. Working to overcome the global impact of neglected tropical diseases, First WHO report on neglected tropical diseases. Geneva, Switzerland: WHO Press; 2010.

Thesis and Dissertations Joselevitch C. Visão no ultravioleta em Carassius

auratus (Ostariophysi, Cypriformes, Cyprinidae): estudo eletrofisiológico do sistema cone - células horizontais. [Master’s dissertation]. São Paulo: Instituto de Psicologia, USP; 1999.

Marcolongo R. Dissolução de medicamentos: fundamentos, aplicações, aspectos regulatórios e perspectivas na área farmacêutica. [dissertação]. São Paulo: Universidade de São Paulo, Faculdade de Ciências Farmacêuticas; 2003.

Laws Agência Nacional de Vigilância Sanitária (Brasil). Resolução

nº. 259, de 20 de setembro de 2002. Regulamento Técnico para Rotulagem de Alimentos Embalados. Diário Oficial da União 23 set 2002; Seção 1.

Conference, Symposium Proceedings. Cite papers only from published proceedings.

Hejzlar RM, Diogo PA. The use of water quality modelling for optimizing operation of a drinking water reservoir. In: Proceedings of the International Conference Fluid Mechanics and Hydrology. 1999 Jun 23-26; Prague. Prague: Institute of Hydrodynamics AS CR; 1999. p 475-482.

Proceedings of the 10th annual meeting of the Canadian Society for Pharmaceutical Sciences. J Pharm Pharm Sci. 2007 Dec 3;10(4):1s-186s.

Audiovisual Material Physician’s Desk Reference (PDR). Release 2003.1AX. [CD-

ROM]. Montvale: Thomson PDR; 2003.

Computer Program Dean AG, Dean JA, Coulombier D, Brendel KA, Smith DC,

Burton AH, et al. Epi info, version 6.04: a word processing database and statistics program for public health on IBM-compatible microcomputers. [Computer program]. Atlanta: Centers of Disease Control and Prevention; 1998.

Patent Larsen CE, Trip R, Johnson CR. Methods for procedures

related to the electrophysiology of the heart. Patent No. 5.529.067. Novoste Corporation; 1995.

“Unpublished results” and “Personal communication”. Reference should appear in the text with the individual name(s) and initials and not in the reference list. (Santos CS, da-Silva GB, Martins LT, unpublished results).

It is assumed that the author has obtained permission from the source when “personal communication” is cited.

HOW TO SUBMIT A MANUSCRIPT TO THE BJPS

Submission should be sent electronically through the ScholarOne system (https://mc04.manuscriptcentral.com/bjps-scielo)

If you need further assistance, please contact the Journal Staff ([email protected]).

mailto:[email protected]

editorial board...v brazilian journal of pharmaceutical sciences the 53th pharmacy and biochemistry...

Documents