Bioprospecção da espécie Miconia willdenowii visando a obtenção de substâncias com potencial antichagásico

Abstract

In this work, ten different ethanol crude extracts were prepared from different plant species of rainforest nearby the city of Alfenas-MG. After standardization of in vitro antichagasical test against T. cruzi (Y strain) epimastigote form, and using colorimetric method with resazurin solution, all extracts had the activity evaluated and the most active was Miconia willdenowii extract, from the Melastomataceae family. This extract was subsequently subjected to partitioning by liquid-liquid extraction using hexane, ethyl acetate and ethanol:water (1:3) mixture, allowing the separation of compounds in groups by difference in polarity. The 3 fractions obtained were submitted to bioguided assay in order to determine the most active. The ethyl acetate fraction displaying the higher potential against epimastigote form of Chagas’ disease parasites was extensively purified under CC conditions. In the end, this process provided a total of 12 subfractions. All subfractions were subjected to bioguided assay, determining those with the greatest potential against epimastigote. Overall, three subfractions showed antichagasical activity (A2, A3 and A4), which were analyzed by high performance liquid chromatography (HPLC), and two hydroquinones derivatives previously reported for other species Miconia: miconidin and primin, could be isolated. Both compounds had their structures identified by NMR spectroscopy, mass spectrometry and confirmed by database comparison in the literature. The quantification of these two compounds was made for each sample of M. willdenowii (crude extract, acetate fraction and subfractions), which allowed deducing that their presence or absence is related to trypanocidal activity. The anti-T. cruzi bioassay, when performed to the isolated compounds, allowed observing higher trypanocidal potential than the reference drug (benznidazole). The in vitro cytotoxicity assay for miconidin and primin, using normal human skin-derived fibroblasts, showed that they only present cytotoxicity over 50 µg/mL, in which is higher than the EC50 presented by these compounds in the trypanocidal assay, suggesting high potential for effective use of these compounds as therapeutical alternative for Chagas’ disease after further studies.