Unraveling putative antiulcer phytoconstituents against Helicobacter pylori urease and human H⁺/K⁺-ATPase from Jacaranda mimosifolia using UPLC-MS/MS coupled to chemometrics and molecular docking

Globally peptic ulcer disease is a severe public health issue caused by an imbalance between the defensive and aggressive mechanisms. In the current study, the phytochemical components of various organs (leaf, fruit, seed, and bark) of Jacaranda mimosifolia D. Don collected from Egypt at different seasons and geographical locations were investigated. UPLC-MS/MS allows a tentative identification of 53 compounds where flavonoids, quinoids, and triterpenoids represented the major identified classes. Multivariate statistical analysis (principle component analysis (PCA), hierarchical cluster analysis (HCA)-heat map and orthogonal projection to latent structuresdiscriminate analysis (OPLS-DA), and accompanied coefficient plots) was applied to investigate in-between and within-class discrimination of the different organs of Jacaranda mimosifolia D. Don. The obtained extracts were tested for their inhibitory activities against Helicobacter pylori Urease and human H+/K+-ATPase enzymes. Fruit extract showed the highest inhibitory activity against H. pylori urease. Meanwhile, leaf extract was found to be more potent, against human H+/K+-ATPase. The results of the biological activities were further modeled using Partial Least Squares Regression (PLSR) analysis together with the quantitative data. The optimal number of latent variables was determined with the aid of Variables important for the projection (VIP) plot to remove the uninformative variables. The developed model was validated using root mean square error of calibration (RMSEC), cross-validation (RMSECV) and prediction (RMSEP). PLS model coefficient plots showed that flavonoids as isoquercitrin and rutin were the most contributing metabolites/biomarkers to the detected anti-urease and H+/K+-ATPase activities, respectively. Molecular docking analysis was utilized to gain more insights into the interaction modalities for the top fifteen metabolites obtained from the PLS model with the active sites of urease and H+/K+-ATPase enzymes. In agreement with the model coefficients, isoquercitrin and rutin were ranked as the top hits for the studied enzymes. To validate the results, in vitro inhibitory assays were performed on isoquercitrin and rutin. Potent inhibitory activities were determined against urease and H+/K+-ATPase with IC50 +/- SD of 9.10 +/- 0.75 mu M and 0.61 +/- 0.06 mu M for isoquercitrin and rutin, respectively.