Isolation, Characterization, and Anti‐Inflammatory Effects of β‐Sitosterol‐β‐D‐Glucoside from Hygrophila auriculata: Experimental Validation, Molecular Docking, and Molecular Dynamics Simulations

Abstract Hygrophila auriculata (K. Schum) Heine is known to treat various common aliment e. g. rheumatoid arthritis, kidney infections, jaundice, edema, and gout. This study aims to isolate bioactive components from the methanolic extract, assess their anti‐inflammatory effects, and investigate their interactions with drug targets through docking and molecular dynamics simulations. Methanolic extract of H. auriculata furnished stigmast‐5‐en‐3‐ol‐β‐D‐glucopyranoside (HA‐06) which was characterized by using IR, NMR and mass spectral data. HA‐06 alleviated carrageenan‐induced inflammation in rats, while the methanolic extract of H. auriculata produced comparable results. The findings were similar to those of the positive control, indomethacin. The chemical structure of HA‐06 was optimized using DFT at the B3LYP level and subsequently used for molecular docking against anti‐inflammatory drug targets. HA‐06 exhibited strong affinity towards phospholipase A2 and glucocorticoid receptor exhibiting binding energies of −11.25 kcal/mol and −11.07 kcal/mol, respectively. Molecular dynamics simulation was used to assess the dynamical stability of these two complexes and their native co‐crystallized ligands. Principal component analysis, radius of gyration, free energy landscapes, solvent‐accessible‐surface‐area, and root‐mean square deviation/fluctuation all indicated stable interactions. Therefore, HA‐06 could be a promising candidate for development into an effective therapy against inflammatory diseases targeting phospholipase A2 and glucocorticoid receptor.