Estudo químico de Vicia faba (FABACEAE): estratégias de desreplicação molecular, isolamento de flavonóides e avaliação da atividade citotóxica
Data
2024-01-26
Autores
Sipoloni, Victor Menezes [UNIFESP]
Orientadores
Veiga, Thiago André Moura [UNIFESP]
Tipo
Dissertação de mestrado
Título da Revista
ISSN da Revista
Título de Volume
Resumo
O potencial de metabólitos secundários em ensaios clínicos associados a várias doenças tem sido relatado por muitos autores. Nos últimos anos, o uso de técnicas hifenadas impulsionou a descoberta de novos produtos naturais bioativos, promovendo estratégias mais eficazes para a investigação de substâncias conhecidas ou desconhecidas. Portanto, propusemos o uso de abordagens analíticas para a desreplicação molecular de substâncias biossintetizadas por Vicia faba (Fabaceae). Para isso, frações obtidas dos extratos etanólicos de diferentes partes do vegetal foram submetidas a ensaios citotóxicos contra linhagens celulares de leucemia humana (Jurkat, Kasumi-1, Raji e K562), e de carcinoma colorretal (HCT-116). Em seguida, através de análises por espectrometria de massas tandem (EM/EM), os dados foram analisados através da plataforma GNPS, para a construção de redes moleculares, a partir das quais foram agrupados os íons detectados através da similaridade dos espectros de massas, onde foi possível realizar anotações de substâncias pertencentes à classe dos flavonóides. Por fim, as frações citotóxicas foram submetidas a diferentes processos de purificação. As doze substâncias isoladas estão sendo caracterizadas por dados de RMN e EMAR, sendo que quatro delas foram identificadas como isoflavonóides, classe de substâncias nunca antes relatada para o gênero Vicia: duas isoflavonas e duas isoflavanonas (inéditas na literatura). As duas isoflavonas não apresentaram atividade citotóxica frente às diferentes linhagens testadas. Por outro lado, as isoflavanonas apresentaram atividade contra a linhagem HCT-116. Esses dados sugerem que a abordagem de desreplicação foi importante para promover a expansão da diversidade química do gênero em estudo, e que, além disso, pode nos oferecer substâncias com efeito citotóxico.
The potential of secondary metabolites in clinical trials associated with various diseases has been reported by many authors. In recent years, the use of hyphenated techniques has boosted the discovery of new bioactive natural products, promoting more effective strategies for the investigation of known or unknown substances. Therefore, we proposed the use of analytical approaches for the molecular dereplication of substances biosynthesized by Vicia faba (Fabaceae). To this end, fractions of the ethanol extract obtained from different parts of the plant were subjected to cytotoxic assays against human leukemia cell lines (Jurkat, Kasumi1, Raji and K562), and colorectal carcinoma (HCT116). Then, using tandem mass spectrometry (MS/MS) analysis, the data was analyzed using the GNPS platform to build molecular networks, from which the ions detected were grouped by mass spectra similarity, where it was possible to make annotations of substances belonging to the flavonoid class. Finally, the cytotoxic fractions were subjected to different purification processes. The twelve isolated substances are being characterized by NMR and HRMS data, four of which have been identified as isoflavonoids, a class of substances never before reported for the genus Vicia: two isoflavones and two isoflavanones (unpublished in the literature). The two isoflavones showed no cytotoxic activity against the different strains tested. On the other hand, the isoflavanones showed activity against the HCT116 strain. These data suggest that our dereplication approach was important in promoting the expansion of the chemical diversity of the genus under study, and that it may also provide us with substances with cytotoxic effects.
The potential of secondary metabolites in clinical trials associated with various diseases has been reported by many authors. In recent years, the use of hyphenated techniques has boosted the discovery of new bioactive natural products, promoting more effective strategies for the investigation of known or unknown substances. Therefore, we proposed the use of analytical approaches for the molecular dereplication of substances biosynthesized by Vicia faba (Fabaceae). To this end, fractions of the ethanol extract obtained from different parts of the plant were subjected to cytotoxic assays against human leukemia cell lines (Jurkat, Kasumi1, Raji and K562), and colorectal carcinoma (HCT116). Then, using tandem mass spectrometry (MS/MS) analysis, the data was analyzed using the GNPS platform to build molecular networks, from which the ions detected were grouped by mass spectra similarity, where it was possible to make annotations of substances belonging to the flavonoid class. Finally, the cytotoxic fractions were subjected to different purification processes. The twelve isolated substances are being characterized by NMR and HRMS data, four of which have been identified as isoflavonoids, a class of substances never before reported for the genus Vicia: two isoflavones and two isoflavanones (unpublished in the literature). The two isoflavones showed no cytotoxic activity against the different strains tested. On the other hand, the isoflavanones showed activity against the HCT116 strain. These data suggest that our dereplication approach was important in promoting the expansion of the chemical diversity of the genus under study, and that it may also provide us with substances with cytotoxic effects.