Metabolômica e redes moleculares no Rastreamento de Substâncias Bioativas de espécies vegetais: Uma Abordagem Estratégica para a Leucemia Mieloide

Abstract

Medicinal plants show a long history of use in traditional folk medicine, standing out for the diversity of properties that are beneficial to human health. They constitute important promising targets for the search for bioactive substances with potential anticancer, anti-inflammatory, antibacterial activities, among others. The Pantanal and Cerrado biomes have a wide biodiversity, sheltering numerous species of medicinal plants, whose chemical and biological properties remain little explored scientifically. In this context, advanced tools, such as metabolomics and molecular networks, have been successfully used to accelerate the discovery and reduce the time required to identify compounds of bioactive metabolites. The main objective of this study was to evaluate the cytotoxic activity of extracts obtained from eighteen plant species found in the Cerrado and Pantanal biomes of Mato Grosso do Sul against Kasumi-1, KG-1 and K562 leukemic cell lines, in addition to applying statistical analysis tools of metabolomic data together with molecular networks to determine the possible bioactive compounds. The plant samples (18 different species) were extracted by accelerated solvent extraction (ASE) using ethanol and water in the proportion 7:3 (v/v) and were analyzed by high-performance liquid chromatography coupled to a diode array detector and a high-resolution mass spectrometer (HPLC-DAD-MS) for the analysis of all extracts. The cytotoxicity of the extracts was evaluated by Alamar Blue assay and the induction of apoptosis was determined by flow cytometry with annexin V and propidium iodide, aiming to identify cells in early and late apoptosis. Metabolomic analyses, complemented by statistical analyses and molecular networks, allowed a comprehensive approach of the metabolites present in the different extracts studied, enabling the identification and analysis, allowing the annotation of steroidal and aporphinic alkaloids as priority targets. The extracts from Sesbania virgata, Aeschynomene denticulata, Erythroxylum anguifugum, Psidium guineense, Astronium fraxinifolium, Coccoloba ochreolata, Solanum glaucophyllum and Paullinia pinnata demonstrated significant inhibition of cell viability of the leukemic cell line K-562 (approximately 70% at 100 µg/mL), while Ocotea diospyrifolia showed 35% inhibition for the KG-1 cell line. Alkaloid fractions isolated from S. glaucophyllum and O. diospyrifolia demonstrated EC50 values ranging from 13.9 to 6.4 μg/mL for the K-562 and KG-1 cell lines, inducing apoptosis and significant cellular damage. The alkaloid boldine presented EC50 values of 46, 116, and 145 µM for the Kasumi-1, KG-1 and K-562 leukemia cell lines, respectively. The results highlight the importance of integrating broader analysis of chemical data for identification of bioactive compounds, emphasizing the potential benefits in the search for metabolites against leukemia cancer cells, particularly steroid and aporphine alkaloids. Therefore, metabolomic strategies and molecular networks for the effective identification of bioactive compounds, highlighting the potential of Cerrado and Pantanal plants for the development of new antileukemic agents.