Estudo de ADME de candidatos a fármacos para o tratamento da leishmaniose visceral e tripanossomíase americana
Data
2019-02-21
Autores
Silva, Bruna Cristina Ulian
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
Estudos precoces de ADME podem agilizar o processo de desenvolvimento de novos fármacos, selecionando os mais promissores para futuros estudos farmacocinéticos. Nesse sentido, e considerando a necessidade de novos tratamentos para a leishmaniose visceral e para a tripanossomíase americana, as moléculas LINS03006, LINS03011 e LINS03013 com ação sobre o Trypanosoma cruzi e a Leishmania infantum foram desenvolvidas por pesquisadores da UNIFESP/Diadema. O presente trabalho teve como objetivo realizar estudos físico-químicos e de ADME para a seleção das moléculas LINS mais promissoras para futuros estudos in vivo. Métodos analíticos e bioanalíticos por UHPLC/UV foram desenvolvidos para a quantificação dos compostos nos ensaios realizados. Foram realizados estudos de estabilidade química em diferentes pHs (1,2; 7,4 e 8,8), estabilidade metabólica em microssomos de rato e humano, estabilidade ex vivo, coeficientes de partição (log P) e permeabilidade em células Caco-2. Os compostos apresentaram estabilidade de até 24 horas em todos os pHs avaliados, levando à expectativa de que na administração oral não ocorrerá degradação decorrente do pH estomacal, sanguíneo e duodenal. O estudo em plasma de rato resultou, para todos os compostos, estabilidade em tempo suficiente – ao menos 6 horas – com expectativa de que ocorra efeito in vivo. Nos estudos de logP, os compostos apresentam características lipofílicas, favorecendo a permeabilidade e o clearance hepático. No ensaio de permeabilidade, os compostos LINS apresentaram Papp> 5,0 x E-06 cm/s, podendo ser classificados como de média ou alta permeabilidade, dessa forma, há expectativa de absorção através do epitélio intestinal. Na estabilidade metabólica em microssomos de ratos, os compostos LINS03006 e LINS03013 demonstraram ser substratos das enzimas do citocromo P450, diferente do LINS03011. Em microssomos humano nenhum dos compostos LINS apresentaram degradação, evidenciando que o clearance hepático não é a principal via de eliminação em humanos. Os resultados obtidos demonstram que as moléculas apresentam características adequadas à continuidade do desenvolvimento.
Early studies of ADME can speed up the development of new drugs, selecting the most promising for future pharmacokinetic studies. In this sense, and considering the need for new treatments for visceral leishmaniasis and american trypanosomiasis, the molecules LINS03006, LINS03011 and LINS03013 with action on Trypanosoma cruzi and Leishmania infantum were developed by researchers from UNIFESP/Diadema. The present work aimed to perform physical-chemical and ADME studies to select the most promising LINS molecules for future in vivo studies. Analytical and bioanalytical methods by UHPLC/UV were developed for the quantification of compounds in the assays performed. Chemical stability studies were performed at different pHs (1.2, 7.4 and 8.8), metabolic stability in rat and human microsomes, ex vivo stability, partition coefficients (log P) and permeability in Caco-2 cells. The compounds exhibited stability of up to 24 hours in all evaluated pHs, leading to the expectation that oral degradation will not occur due to stomach, blood and duodenal pH. The study in rat plasma resulted, for all compounds, stability in sufficient time - at least 6 hours - with the expectation that in vivo effect would occur. In logP studies, the compounds presented lipophilic characteristics, favoring permeability and hepatic clearance. In the permeability assay, the LINS compounds presented Papp > 5.0xE-06 cm/s, which can be classified as medium or high permeability, thus, there is an expectation of absorption through the intestinal epithelium. In the metabolic stability in microsomes of rats, compounds LINS03006 and LINS03013 were shown to be substrates of cytochrome P450 enzymes, different from LINS03011. In human microsomes none of the LINS compounds showed degradation, evidencing that hepatic clearance is not the main route of elimination in humans. The results obtained demonstrate that the molecules present characteristics suitable for the continuity of development.
Early studies of ADME can speed up the development of new drugs, selecting the most promising for future pharmacokinetic studies. In this sense, and considering the need for new treatments for visceral leishmaniasis and american trypanosomiasis, the molecules LINS03006, LINS03011 and LINS03013 with action on Trypanosoma cruzi and Leishmania infantum were developed by researchers from UNIFESP/Diadema. The present work aimed to perform physical-chemical and ADME studies to select the most promising LINS molecules for future in vivo studies. Analytical and bioanalytical methods by UHPLC/UV were developed for the quantification of compounds in the assays performed. Chemical stability studies were performed at different pHs (1.2, 7.4 and 8.8), metabolic stability in rat and human microsomes, ex vivo stability, partition coefficients (log P) and permeability in Caco-2 cells. The compounds exhibited stability of up to 24 hours in all evaluated pHs, leading to the expectation that oral degradation will not occur due to stomach, blood and duodenal pH. The study in rat plasma resulted, for all compounds, stability in sufficient time - at least 6 hours - with the expectation that in vivo effect would occur. In logP studies, the compounds presented lipophilic characteristics, favoring permeability and hepatic clearance. In the permeability assay, the LINS compounds presented Papp > 5.0xE-06 cm/s, which can be classified as medium or high permeability, thus, there is an expectation of absorption through the intestinal epithelium. In the metabolic stability in microsomes of rats, compounds LINS03006 and LINS03013 were shown to be substrates of cytochrome P450 enzymes, different from LINS03011. In human microsomes none of the LINS compounds showed degradation, evidencing that hepatic clearance is not the main route of elimination in humans. The results obtained demonstrate that the molecules present characteristics suitable for the continuity of development.
Descrição
Palavras-chave
Leishmaniose;, Tripanossomíase, ADME, Caco-2, Microssomos., Microsomes