Perfil metabolômico e potencial citotóxico de Phyllanthus spp. (Phyllanthus amarus Schum. & Thonn. e P. niruri L.)

• Ano de defesa: 2019
• Autor(a) principal: Silva, Maria Francilene Souza
• Tipo de documento: Tese
• Tipo de acesso: Acesso aberto
• Idioma: por
• Palavras-chave em Português: Desreplicação; Metabolômica; Quebra-pedra; Citotoxicidade; Câncer
• Link de acesso: http://www.repositorio.ufc.br/handle/riufc/41644

The genus Phyllanthus belongs to Phyllanthaceae family, including the species Phyllanthus amarus and P. niruri, known as “quebra-pedra”. The use of natural products and its new chemical structures to develop new drugs are in focus the last years. New strategies as dereplication aim to fast the identification of the compounds from a crude extract or partial purified. The current study aims to evaluate the technological potential of P. amarus and P. niruri investigating the influence of solid phase extraction (SPE) under its cytotoxic and chemical profile. The studies were performed using UPLC-QTOF-MSE and RMN of aqueous extracts of the two species, as well as fractionation by preparative HPLC, correlating the chemical profiles of the fractions with biological activity. The scientific and technological monitoring was carried out based on articles found in periodicals in the Web of Science e NCBI (PUBMED) and patent basis ESPACENET, GooglePatents and INPI. The analysis of scientific and technological monitoring demonstrated the pharmacological potential of P. amarus and P. niruri as agents for the treatment of different types of diseases and possibilities of clinical trials in several health areas. The SPE was performed with Oasis, Plexa and Agilent C18 cartridges with and without organic modifiers. As results showed differences between the SPE cartridges and the recovered mass, allowed the exploration of the different chemical functionalities of the Phyllanthus species. The SPE samples showed higher in vitro cytotoxicity than the crude extracts. Preparative HPLC was used for fractionation of the extracts and the chemical composition was determined by UPLC-QTOF-MSE and NMR, together with chemometric techniques (PCA and PLS). Biological activity was tested on a panel of tumor and non-tumor cell lines by the MTT assay and flow cytometric assays were performed, such as cell proliferation assays, membrane integrity, cell cycle, mitochondrial transmembrane potential and mechanism of death. The variability of the chemical composition of the fractions showed that fractions 4 and 5 of the two species analyzed by UPLC-QTOF-MSE and NMR showed higher content of chemical compounds. The PhyN Fr. 4n fraction (P. niruri fraction 4) showed higher cytotoxic activity against the HL60 line and the cytometric tests showed that after 24 h of treatment the PhyN Fr. 4n fraction presented progressive reduction of the number of cells at the tested concentrations and reduced cell viability in a dependent concentration. In the evaluation of cell death, PhyN Fr. 4n at concentrations of 47 and 70.5 μg / mL showed a significant increase of cells in late apoptosis. The PhyN Fr. 4n fraction binds cells in the S phase to concentrations of 47 and 70.5 μg / ml. Morphologically the treated cells presented apoptosis and / or necrosis death characteristics. The fraction 4n present as metabolites of greater intensity of repandusinic acid A-isomer I (1) and II (3) , brevifolin carboxylic acid (2), corilagin (4), corilagin-isomer (5) and geraniin- isomer II (16). We can verify in this study that the species of Phyllanthus present great chemical and pharmacological potential to be explored regarding the research with anticancer drugs.