Chemometrics applied to metabolomics of Aldama La Llave: chemotaxonomic contributions and targeted phytochemistry based on cyclooxygenase-1 and 5-lipoxygenase inhibition

The species of the genus Viguiera Kunth, recently transferred to the genus Aldama La Llave (Asteraceae, Heliantheae), still show problems of taxonomic boundaries. This genus has 35 species distributed throughout the Brazilian territory, mostly in the cerrado. Chemical analyses has demonstrated its potential to help in the solution of problems at various taxonomic levels based on chemical groups of secondary metabolites. Furthermore, phytochemical analyses have shown that certain compounds from Aldama have the potential to be studied aiming to investigate anti-inflammatory molecular mechanisms. Thus, in this work it was proposed to carry out the untargeted metabolomic approach aided by chemometrics, aiming to provide chemical data either for chemotaxonomic contributions and to find bioactive compounds. Through this approach, both unsupervised (PCA and HCA) and supervised multivariate analysis (OPLS-DA) combined with data from UHPLC-DAD-Orbitrap analyses showed that the genus Aldama is \"chemoconsistent\" and thus certain discriminant compounds were suggested. Additionally, some species with taxonomic problems had their infrageneric positioning explained from the chemotaxonomic point of view of. With regards to the analyses carried out to find bioactive compounds (targeted phytochemistry), the inflammatory targets investigated in this study were the COX-1 and LOX-5 pro-inflammatory enzymes with which in vitro assays were made and positive activity of extracts from species of Aldama was observed. A. trichophylla was the selected species for the targeted phytochemistry because it showed dual inhibition activity against both enzymes and it is still little investigated from the chemical point of view. The best models for inhibition of COX-1 and 5-LOX obtained with chemical data of Aldama were selected by means of various combinations of algorithms, processing software (MZmine, MetAlign and MSClust) and hyphenated techniques, such as HPLC-TOF and UHPLC-Orbitrap. Thus, certain peak ions were appointed by chemometrics as discriminant for the dual inhibition activity. These peaks were dereplicated based on their UV profiles and fragmentation patterns via HCD-Orbitrap and Ion-Trap. Thus, it was possible to dereplicate three probable chemical inhibitors of COX-1 and 5-LOX: kaempferol-3-O-glucuronide, quercetin-3-O-methyl-7-glucuronide and kaempferol-3-O-(6\"-malonyl-glucoside). Finally, the targeted phytochemistry was carried out. The chromatographic fractionation allowed us to isolate these three compounds and a preliminary 1H NMR analysis was performed in order to conclude their structural identification. In the near future, these substances will have their activity evaluated by the inhibition of the two enzymes. (AU)