RESUMO
Doenças tropicais negligenciadas atingem cerca de 1,7 bilhões de pessoas, gerando um forte impacto na economia e problemas na Saúde Pública. Dentre as endemias mais negligenciadas, encontra-se a doença de Chagas, que afeta cerca de 6 milhões de pessoas, e no Brasil, dispõe-se de apenas de um fármaco altamente tóxico contra a infecção. Sendo assim, existe uma necessidade urgente para novos tratamentos. A exploração farmacológica de compostos produzidos por microrganismos é de longa data e contribuiu até hoje, com diversos fármacos aprovados. O presente projeto avaliou o potencial anti-Trypanosoma cruzi de metabólitos de espécies bacterianas marinhas encontradas no litoral paulista. Para isto, foram coletados invertebrados e sedimentos marinhos e isoladas 32 espécies bacterianas, resultando em 12 microrganismos identificados por MALDI-TOF/MS ou sequenciamento genético. Os extratos orgânicos, contendo os metabólitos microbianos, foram avaliados quanto ao potencial anti-T. cruzi em tripomastigotas, apresentando valores de Concentração Efetiva 50% (CE50) entre 1,5 e 59,0 µg/mL. Duas cepas foram submetidas à abordagem One Strain Many Compounds (OSMAC), porém, não se observou aumento da potência antiparasitária. O pré-fracionamento do extrato da Olleya marilimosa, resultou em uma fração (FII) ativa contra os tripomastigotas (CE50 23 µg/mL), com ausência de citotoxicidade em fibroblastos e hemácias até 200 µg/mL. A análise em ressonância magnética nuclear (RMN 1H) e espectrometria de massas de alta resolução, demonstrou a presença de 4 ácidos graxos de cadeia iso, 1 (C19H37O2), 2 (C20H39O2), 3 (C21H41O2) e 4 (C22H43O2)...(AU)
Neglected tropical diseases affects about 1.7 billion people, generating a strong impact on the economy and problems in Public Health. Among the most neglected endemic diseases is Chagas disease, which affects about 6 million people, and in Brazil, only one highly toxic drug is available against it. Therefore, there is an urgent need for new treatments. The pharmacological exploitation of compounds produced by microorganisms is long-standing and has contributed to several approved drugs. The present project evaluated the anti-Trypanosoma cruzi potential of metabolites of marine bacterial species found on the coast of São Paulo. For this, invertebrates and marine sediments were collected and 32 bacterial species were isolated, resulting in 12 microorganisms identified by MALDI-TOF/MS or genetic sequencing. The organic extracts containing the microbial metabolites were evaluated for antiT. cruzi potential. crossed in trypomastigotes, presenting Effective Concentration 50% (EC50) between 1.5 and 59.0 µg/mL. Two strains were submitted to the One Strain Many Compounds (OSMAC) approach, however, there was no increase in antiparasitic potency. Prefractionation of Olleya marilimosa extract resulted in an active fraction (FII) against trypanomastigotes (EC50 23 µg/mL), with no cytotoxicity in fibroblasts or red blood cells up to 200 µg/mL. Nuclear Magnetic Resonance analysis (NMR 1H) and High-Resolution Mass Spectrometry demonstrated the presence of 4 iso chain fatty acids, 1 (C19H37O2), 2 (C20H39O2), 3 (C21H41O2) and 4 (C22H43O2). Using spectrofluorimetry, it was observed that FII induced a change in the permeability of the plasma membrane of the parasite. Analysis in mass spectrometry (MALDI-TOF/MS) also demonstrated changes in the protein profile of parasites after treatment. This study presented in an unprecedented way, the anti-T. cruzi potential metabolites of the marine bacteria studied. The isolation and characterization of these compounds may contribute to new pharmaceutical prototypes for Chagas disease. (AU)