Inhibitory Mechanisms of <i>β</i> ‐Glucuronidase by <i>Hibiscus syriacus</i> Phenolics: Integrating Computational and Experimental Approaches
Haifa A. Alqhtani, Sarah I. Othman, Faris F. Aba Alkhayl, Naif G. Altoom, Al Mokhtar Lamsabhi, Emadeldin M. Kamel
Abstract
Abstract Exploring the intricate mechanisms of β ‐glucuronidase inhibition is essential to advancing the development of novel therapeutic agents. This study extensively evaluated the inhibitory potential of phenolics from Hibiscus syriacus against β ‐glucuronidase using a combination of in vitro and computational approaches. in vitro assays demonstrated that chlorogenic acid and dactylifric acid exhibited significant inhibitory activity, with low IC 50 values of 1.32 ± 0.08 and 10.02 ± 1.38 µM, respectively. Enzyme kinetics analyses revealed that dactylifric acid and the positive control, EGCG, followed a mixed inhibition mechanism, while chlorogenic acid displayed competitive inhibition, as indicated by the intersecting lines in the Lineweaver–Burk plots. Docking studies supported these in vitro findings, with chlorogenic acid and dactylifric acid showing the lowest binding affinities, extensive polar interactions, and occupancy of identical binding sites as the reference drug. A 30 ns molecular dynamics simulation was performed to explore the interaction dynamics between isolated phenolic compounds and β ‐glucuronidase. Evaluation of multiple MD parameters revealed that chlorogenic acid and dactylifric acid exhibited stable trajectories and substantial energy stabilization in their binding with β ‐glucuronidase. These computational findings are consistent with experimental data, supporting chlorogenic acid and dactylifric acid as potential inhibitors of β ‐glucuronidase.