Search

Agradecimentos: This study was funded by Biolab Indústria Farmacêutica Ltd., Brazil and Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP - Grant n° 2016/22506-1) Abstract: Nanotechnology may increase the speed of penetration into the skin. This study evaluated the efficacy, safety, and pharmacokinetics of a novel topical anesthetic nanocapsule formulation (2 g) containing 2.5% lidocaine and 2.5% prilocaine (nanorap - test formulation) compared to placebo (control formulation) in skin types I-III patients of both sexes submitted to the ablative fractional CO2 laser treatment.The patients (n = 120) included in this double-blind, single-center, randomized trial, received topical application of 2 g of the test formulation (50 mg lidocaine + 50 mg prilocaine) and placebo on the forehead region. Efficacy was assessed as pain sensation in four quadrants of each side of the forehead using a visual analogue scale immediately (0?min) and at 30, 60, and 90 minutes after laser application compared to placebo. The safety and tolerability of the test product were evaluated based on the occurrence of systemic adverse events as well as the occurrence of immediate and late skin reactions. Pharmacokinetic evaluation was performed in plasma of eight patients using a validated LC-MS/MS method for drugs quantification.Nanorap induced a clinically significant reduction in the pain assessment at all evaluated times (57.2%, 41.6%, 38.6%, and 37.3% at 0, 30, 60, and 90 minutes after drug application, respectively. Mean values of Cmax were 14.20 and 5.36?ng/ml and tmax were 3.5 and 1.8 hour for lidocaine and prilocaine, respectively. No systemic adverse events were observed.The nanorap formulation demonstrated a clinically and statistically significant efficacy providing analgesia after the ablative fractional CO2 laser therapy in the investigated patients, when compared to placebo. The product also presented good safety and tolerability FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP Fechado

Agradecimentos: This study was funded by Biolab Sanus Farmacêutica Ltda., Brazil Abstract: The combination of medroxyprogesterone acetate 25 mg + estradiol cypionate 5 mg is a highly effective, monthly injectable contraceptive. For the first time, this study presents the development and validation of a sensitive method for estradiol cypionate analysis in human plasma by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). Aliquots (500 µL) of plasma were extracted with ethyl ether (100%) and derivatized with dansyl chloride. Its separation was performed on a Jones Chromatography Genesis C8 column and the quantification was performed with a mass spectrometer equipped with an electrospray interface operating in negative ion mode. The run time was 6 min and the calibration curve was linear over the range of 0.005-0.15 ng/mL. The method was applied to evaluate the pharmacokinetics of estradiol cypionate in plasma collected up to 1008 h (42 days) after a single intramuscular administration of 25 mg/mL medroxyprogesterone acetate +5 mg/mL estradiol cypionate to healthy female volunteers (n = 12). The estradiol cypionate maximum plasma concentration (Cmax) was 0.14 ± 0.08 ng/mL reached at 16.83 ± 21.07 h and the area under the plasma concentration versus time curve (AUC0-last) was 14.07 ± 6.32 ng.h/mL. Elimination half-life (t½), apparent volume of distribution (Vd/F), apparent clearance (CL/F) and mean residence time (MRT) were 89.65 ± 76.04 h, 28038 ± 9636 L, 49.02 ± 10.62 L/h and 576.05 ± 238.32 h, respectively, showing that the estradiol cypionate release from the administration site was prolonged and there was no drug accumulation Fechado

Agradecimentos: This work was supported by the Brazilian Ministry of Science and Technology and the National Council for Scientific and Technological Development through the public announcement PQ10/2011 - productivity in research - PQ-2011 (grant number 309228/2011-5) Abstract: Bariatric surgery leads to several anatomo-physiological modifications that may affect pharmacokinetic parameters and consequently alter the therapeutic effect of drugs, such as antibiotics. The pharmacokinetics of oral amoxicillin after Roux-en-Y gastric bypass (RYGB) surgery is unknown.The objective of this study was to evaluate the impact of bariatric surgery on the pharmacokinetics of amoxicillin.This study was performed as a randomized, open-label, single-dose clinical trial, with two periods of treatment, in which obese subjects (n = 8) received an amoxicillin 500 mg capsule orally before and 2 months after the RYGB surgery. The amoxicillin plasma concentration was determined by liquid chromatography coupled to mass spectrometry (LC-MS/MS).After the surgery, the mean weight loss was 17.03 ± 5.51 kg, and mean body mass index (BMI) decreased from 46.21 ± 2.82 to 38.82 ± 3.32 kg/m2. The mean amoxicillin area under the plasma concentration versus time curve from time zero to the time of the last quantifiable concentration (AUC0–tlast) increased significantly (3.5-fold); the maximum plasma concentration (Cmax) increased 2.8-fold after the bariatric surgery. No correlation was found between amoxicillin absorption, BMI, and weight loss percentage.The alterations observed in the amoxicillin pharmacokinetics suggest that obese subjects included in this trial had a substantially increase in amoxicillin systemic exposure after RYGB surgery. However, despite this increase, its exposure was lower than the values reported for non-obese volunteers CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQ Fechado

Orientador: José Luiz da Costa Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas Resumo: O aparecimento de Novas Substâncias Psicoativas (NSP) no mercado ilícito mundial cresceu significativamente desde os anos 2000. Apesar da queda no número de novas identificações anualmente, elas ainda causam severos casos de intoxicações, diversos deles fatais. O presente trabalho desenvolveu e validou um método para a análise de mais de 100 NSP e drogas de abuso convencionais em amostras de fluido oral (OF) e analisado por cromatografia líquida acoplada a espectrometria de massas sequencial (LC-MS/MS). Foram coletadas 462 amostras em festas e festivais de música eletrônica em 3 estados brasileiros e analisadas com a metodologia desenvolvida. O 3,4-metilenodioximetanfetamina (MDMA, 88%) ainda foi a substância mais identificada nas amostras, mas 39% das amostras continham alguma NSP. Cetamina (75%) foi a mais prevalente, seguido da metilona (15%) e N-etilpentilona (10%). Ainda assim, apenas 5% dos voluntários haviam reportado o consumo de alguma NSP. A ingestão de múltiplas substâncias foi muito prevalente, com 80% das amostras contendo duas ou mais substâncias. Este foi o primeiro dado publicado no Brasil contendo dados epidemiológicos do consumo de NSP através da análise de amostras biológicas em contexto recreacional. As amostras contendo catinonas sintéticas foram quantificadas usando microextração por sorvente empacotado (MEPS). Entre as 41 amostras, metilona, N-etilpentilona, eutilona, mefedrona e 4-CEC foram quantificadas. As amostras contendo LSD foram quantificadas, assim como seu principal produto de biotransformação, 2-oxo-3-hidroxi-LSD, com limite de detecção de 0,01 ng/mL. Este foi o primeiro dado publicado na literatura contendo quantificação de ambos analitos em amostras autênticas de fluido oral. Ainda, como forma de suprir a necessidade de obter mais informações sobre as NSP, foi realizado o ensaio de metabolismo in vitro do canabinóide sintético ADB-4en-PINACA usando microssomas hepáticos humanos (HLM) e analisado com espectrômetro de massas de alta resolução. Dos 11 metabólitos identificados in vitro, 7 foram confirmados em amostras de sangue e urina post mortem., sendo os metabólitos derivados de hidroxilação e hidroxilação seguida de hidrólise interna os mais intensos Abstract: New Psychoactive Substances (NPS) emergence of in the global illicit market has grown significantly since the 2000s. Despite the drop in the number of new identifications annually, they still cause severe cases of intoxication, many of them fatal. The present work developed and validated a method for the analysis of more than 100 NPS and conventional drugs of abuse in oral fluid (OF) samples and analyzed by liquid chromatography?tandem mass spectrometry (LC-MS/MS). 462 samples were collected at parties and electronic music festivals in 3 Brazilian states and analyzed using the developed methodology. 3,4-methylenedioxymethamphetamine (MDMA, 88%) was still the most identified substance in the samples, but 39% of them contained some NPS. Ketamine (75%) was the most prevalent, followed by methylone (15%) and N-ethylpentylone (10%). Even so, only 5% of the volunteers had reported some NPS consumption. Ingestion of multiple substances was very prevalent, with 80% of samples containing two or more substances. This was the first data published in Brazil containing epidemiological data on NPS consumption through the analysis of biological samples in a recreational context. Samples containing synthetic cathinones were quantified using microextraction by packed sorbent (MEPS). Among the 41 samples, methylone, N-ethylpentylone, euthylone, mephedrone and 4-CEC were quantified. Samples containing LSD were quantified, as well as its main biotransformation product, 2-oxo-3-hydroxy-LSD, with limit of detection of 0.1 ng/mL. This was the first published data quantifying both analytes in authentic oral fluid samples. Also, to meet the need for more information regarding NPS, an in vitro metabolism assay of the synthetic cannabinoid ADB-4en-PINACA was performed using human liver microsomes (HLM) and analyzed with a high-resolution mass spectrometer. Of the 11 metabolites identified in vitro, 7 were confirmed in post mortem blood and urine samples, with the metabolites derived from hydroxylation and hydroxylation followed by internal hydrolysis being the most intense Doutorado Farmacologia Doutora em Farmacologia FAPESP 2018/11849-0 CAPES 88887.571786/2020-00

Orientador: Paulo César Pires Rosa Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas Resumo: O desempenho biológico de um fármaco administrado por via oral pode ser influenciado por diferentes fatores, como as condições fisiológicas e as propriedades físico-químicas do fármaco, em especial a solubilidade. Esse último parâmetro possui relação com o empacotamento das moléculas na estrutura sólida. Técnicas que alteram a organização do estado sólido, como a cocristalização, podem ser empregadas para aprimorar propriedades físico-químicas de fármacos. Cocristais farmacêuticos são formados por um ingrediente farmacêutico ativo (IFA) e um coformador (substância reconhecida como segura, podendo ou não ter atividade farmacológica), que interagem por meio de ligações não covalentes, em proporção estequiométrica definida. Dentre as propriedades aprimoradas por meio da cocristalização estão a solubilidade, a taxa de dissolução, a permeabilidade e a biodisponibilidade. A genfibrozila (GEN) é um fibrato usualmente empregado no tratamento da hipertrigliceridemia, no entanto, estudos recentes abordam seu efeito farmacológico também no tratamento da doença de Alzheimer. Ela possui baixa solubilidade e alta permeabilidade (categoria II do Sistema de Classificação Biofarmacêutica), característica que pode afetar sua absorção. A GEN possui em sua estrutura molecular o grupo ácido carboxílico, capaz de interagir por meio de ligações não covalentes com um coformador ácido, como o ácido trans-cinâmico (ATC), formando homossíntons supramoleculares, resultando no cocristal. Desta forma, este trabalho teve como objetivo sintetizar cocristal de GEN, usando um coformador ácido e caracterizá-lo; avaliar sua solubilidade na faixa de pH fisiológico (1,2; 4,5; 6,8; 7,4) e em meio biorrelevante, além de verificar a viabilidade de células Caco-2 expostas ao cocristal sintetizado, nas concentrações de 10 µM, 50 µM e 100 µM. As metodologias empregadas na síntese do cocristal foram a evaporação lenta de solvente e a moagem assistida por líquido. A caracterização foi realizada por espectroscopia de infravermelho, calorimetria exploratória diferencial, difração de raios X do pó e análise termogravimétrica. A solubilidade foi determinada utilizando a metodologia shake flask e as amostras foram quantificadas por método cromatográfico analítico devidamente validado. Já a viabilidade celular foi avaliada através do ensaio de citotoxicidade, utilizando o reagente MTT. A GEN associada ao ATC, numa proporção equimolar 1:1, foi capaz de formar o cocristal farmacêutico. No entanto, esta nova forma cristalina se mostrou menos solúvel que o fármaco puro. Este resultado é assegurado pela avaliação sistemática do método cromatográfico que comprovou que o método é linear no intervalo de concentrações utilizadas, seletivo, preciso e exato. Apesar do não incremento na solubilidade, o cocristal GEN-ATC não reduziu a viabilidade de células Caco-2, nas concentrações testadas. Abstract: The biological performance of a drug orally administered can be influenced by different factors, such as the physiological conditions and the physical-chemical properties of the drug, especially the solubility. This last parameter is related to the packaging of molecules in the solid structure. Techniques that change the organization of the solid state, such as co-crystallization, can be used to improve the physicochemical properties of drugs. Pharmaceutical co-crystals are formed by an active pharmaceutical ingredient (API) and a coformer (substance recognized as safe, with or without pharmacological activity), which interact through non covalent bonds, in a defined stoichiometric proportion. Some properties are enhanced by co-crystallization such as solubility, dissolution rate, permeability and bioavailability. Gemfibrozil (GEN) is a fibrate used to treat hypertriglyceridemia; however, recent studies have also addressed its pharmacological effect in the treatment of Alzheimer's disease. GEN has low solubility and high permeability (category II of the Biopharmaceutical Classification System), a characteristic that can affect its absorption. The molecular structure of GEN contains the carboxylic acid group, capable of interacting through non-covalent bonds with an acid coformer, such as trans-cinnamic acid (TCA), forming supramolecular homosyntons, resulting in the co-crystal. In this way, the aim of this study was to synthesize GEN co-crystals using an acid conformer, to characterize the co-crystals, to evaluate its solubility in the physiological pH range (1,2; 4,5; 6,8; 7,4) and in a biorelevant medium, and to check the viability of Caco-2 cells exposed to the synthesized co-crystal, in concentrations 10 µM, 50 µM and 100 µM. The methodologies used to synthesize co-crystal were slow solvent evaporation and liquid-assisted grinding. The characterization was carried out by infrared spectroscopy, differential scanning calorimetry, powder X-ray diffraction and thermogravimetric analysis. The solubility was assessed using the shake flask methodology and the samples were quantified by a duly validated analytical chromatographic method. Cell viability was verified through the cytotoxicity assay, using the MTT reagent. GEN associated with TCA, in an equimolar ratio 1:1, was able to form the pharmaceutical co-crystal. However, this new crystalline form showed to be less soluble than the pure drug. This result is guaranteed by the systematic evaluation of the chromatographic method, which proved that the method is linear, selective, precise and exact in the range of concentrations used. Despite of not increasing solubility, the GEM-TCA co-crystal did not reduce the viability of Caco-2 cells at the tested concentrations. Mestrado Farmacologia Mestra em Farmacologia CNPQ 130444/2019-7

Orientadores: Gilberto De Nucci, Natalicia de Jesus Antunes Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas Resumo: O riluzole é um anticonvulsivante benzotiazol usado no tratamento de pacientes com esclerose lateral amiotrófica e está sendo investigado para uso clínico em pacientes com lesão medular.O presente estudo avaliou a farmacocinética e a segurança do neuroprotetor riluzole em cães sadios da raça beagle, após administração intravenosa e por via oral. As doses intravenosas foram de 1,0 e 5,0 mg/kg e as doses orais foram de 1,5, 5,0, 15,0 e 50,0 mg/kg de riluzole. Após a administração, amostras de sangue foram coletadas no intervalo de 0 a 24 h. Foram realizados exames clínicos e laboratoriais para avaliação da segurança e tolerabilidade. O riluzole foi quantificado por cromatografia líquida de alta eficiência acoplada à espectrometria de massa em tandem (LC-MS/MS). Todos os animais apresentaram parâmetros clínicos e laboratoriais dentro da normalidade. A meia- vida de eliminação (t½) após administração intravenosa foi de 2,6 e 2,2 h, a depuração (Cl) foi de 4,3 e 8,6 L/h/kg e o volume de distribuição (Vd) foi de 17,6 e 27,8 L/kg para as concentrações de 1,0 e 5,0 mg/kg do riluzole, respectivamente. A farmacocinética após administração oral foi linear de 1,5 a 15 mg/kg. A concentração plasmática máxima (Cmax) foi de 342,47, 1376,04, 3515,06 e 4475,38 ng/mL, e a t½ foi de 2,16, 1,5, 1,8 e 3,0 h após a administração oral de 1,5, 5,0, 15 e 50 mg/kg de riluzole, respectivamente. Em conclusão, o método se mostrou sensível, preciso, e seletivo para a quantificação do riluzole em plasma de cães da raça beagle. A farmacocinética do riluzole foi linear até a dose de 15 mg/kg e a t½ foi significante menor que os valores relatados em humanos. Além disso, o riluzole foi bem tolerado pelos animais investigados, sem ocorrência de eventos adversos durante o estudo Abstract: Riluzole is a benzothiazole anticonvulsant used in the treatment of patients with amyotrophic lateral sclerosis and it is being investigated for clinical use in patients with spinal cord injury. The present study evaluated the pharmacokinetics and safety of the neuroprotector riluzole in healthy beagle dogs after intravenous and oral administration. The intravenous administered doses were 1.0 and 5.0 mg/kg and the oral doses were 1.5, 5.0, 15.0 and 50.0 mg/kg riluzole. After administration, blood samples were collected from 0 to 24 h. Clinical evaluation and laboratory tests were performed to assess safety and tolerability. Riluzole was quantified by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). All animals presented clinical parameters and laboratorial test results within normality. After intravenous administration of 1.0 and 5.0 mg/kg riluzole, the elimination half-life (t½) values were 2.6 and 2.2 h, clearance (Cl) were 4.3 and 8.6 L/h/kg and volume of distribution (Vd) was 17.6 and 27.8 L/kg, respectively. The pharmacokinetics was linear from 1.5 to 15 mg/kg after oral administration. The maximum plasma concentration (Cmax) was 342.47, 1376.04, 3515.06 and 4475.38 ng/mL, and the t½ was 2.16, 1.5, 1.8 and 3.0 h after oral administration of 1.5, 5.0,15 and 50 mg/kg riluzole, respectively. In conclusion, the method was sensitive, accurate, and selective to quantify riluzole in plasma of beagles. The riluzole pharmacokinetics was linear up to 15 mg/kg and its t½ was significant shorter than in humans. Furthermore, riluzole was well tolerated by the investigated animals, without the occurrence of adverse events during the study Doutorado Farmacologia Doutora em Farmacologia CAPES

Orientadores: Paulo César Pires Rosa, Gilberto de Nucci Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas Resumo: Os agrotóxicos são produtos utilizados no controle de fauna e flora que podem interferir negativamente na produção agrícola. No Brasil há uma vasta produção de grãos, tais como milho, soja e feijão e que são utilizados agrotóxicos com o intuito de melhorar e garantir a produção dos mesmos. Entretanto, sabe-se que a exposição a longo prazo com os agrotóxicos pode determinar impactos à saúde, como possíveis intoxicações. Desta forma, considerando os riscos ambientais e saúde, é de supra importância o desenvolvimento de metodologias eficazes e seguras para avaliar e determinar o nível de exposição de agricultores e pecuaristas em contato direto com agrotóxicos. O objetivo do presente trabalho foi desenvolver uma metodologia analítica para a determinação de glifosato em amostras de urina por meio de Cromatografia Líquida de Alta Eficiência com detecção por fluorescência (CLAE-FL). O procedimento de extração das amostras foi realizado por cartuchos de extração de fase sólida (SPE), apresentando recuperações entre 69,7 e 86,9%. A metodologia analítica foi validada, avaliando-se sua linearidade, precisão, exatidão, sensibilidade e seletividade. A faixa de trabalho estava compreendida entre 1,15 e 120,0 ng/mL. O método é linear com coeficiente de determinação 0,9993. O limite de detecção foi de 0,34ng/mL e o limite de quantificação 1,15ng/mL. O método utilizado na análise de 90 amostras de urina provenientes de trabalhadores rurais que tiveram exposição e contato direto ao pesticida glifosato na região de Nova Mutum-MT, onde foram coletadas amostras no período pré, durante e pós exposição. Em todas as amostras foram encontrados resíduos de glifosato no dia da exposição, com faixa de concentração de 1,65 ng/mL e 7,13ng/mL. É importante ampliar as possibilidades de análise em outras matrizes ambientais e humanas (solo, ar, sangue) para auxiliar em uma avaliação mais ampla de possíveis efeitos sobre a saúde da população e sobre o meio ambiente naquela região. Os dados deste estudo são de grande relevância para o planejamento de estratégias de biomonitoramento na região, visto que eventuais exposições humanas podem gerar impactos negativos na saúde de trabalhadores rurais e outros membros da comunidade, bem como possíveis intoxicações Abstract: Pesticides are intended to control pest, weeds and plant diseases carriers that can negatively interfere on agricultural production. In Brazil, pesticides have often been used to improve and ensure its vast production of grains (e.g. corn, soybeans and beans), however, the pesticide exposure can lead to acute poisoning or even long-term health effects. Therefore, the study and development of new methodologies for the evaluation and determination of the exposure level has become crucial, especially for those who have direct contact with such chemicals. In view of the environmental and health risks, the aim of the present work was the development of an analytical methodology for the glyphosate determination in urine samples thought a High-Performance Liquid Chromatography with fluorescence detection (HPLC-FL) approach. The sample extractions were performed using continuous phase extraction cartridges (SPE) and presented a set recovery between 69.7 and 86.9%. The analytical methodology was validated considering its linearity, precision, accuracy, sensitivity and selectivity: the method presented a good linearity with determination coefficient of 0.993, the working range of the assay was from 1.15 to 120.0 ng / mL, detection limit was 0.34 ng/mL and the quantification limit was 1.15 ng / mL. A total of 90 urine samples were collected from farm workers of Nova Mutum (Mato Grosso State, and from Brazil) before, during and after the exposure/direct contact with glyphosate; all after-exposition sample measurements indicated pesticide residues ranged from 1.65 ng / mL to 7.13 ng / mL. To improve and expand the knowledge about the impact of pesticides in that region, it is essential, in future works, to broaden the scope of analysis to other human and environmental sources such as soil, air and blood. The results obtained in this work showed great relevance for the development of a biomonitoring strategy, once the exposure of farm workers and other members of the community to pesticide can generate negative health impacts Mestrado Farmacologia Mestra em Farmacologia CAPES 01-P-3371/2017 CNPQ FAEPEX