Harnessing Traditional Medicine for Modern Inflammation Targets: Computational Discovery of Oldenlandia diffusa as a Source of Predicted sEH Inhibitors With Validated Anti-Inflammatory Effects in Vitro

Abstract

Objective Chronic inflammation is a pathological basis for numerous diseases, and soluble epoxide hydrolase (sEH) has emerged as a promising target for anti-inflammatory intervention. This study aimed to identify potential sEH inhibitors derived from the traditional Chinese medicinal herb Oldenlandia diffusa through integrated computational and experimental methodologies. Methods A pharmacophore model was constructed from 41 known sEH inhibitors and utilized to virtually screen 948 metabolites of Oldenlandia diffusa. The identified hits underwent molecular docking, ADMET prediction, induced-fit docking, and 50 ns molecular dynamics simulation. The anti-inflammatory activity of the selected compounds was assessed in LPS-stimulated RAW264.7 macrophages by measuring levels of NO and TNF-α. Results Two compounds, Syringaresinol and 3,7-Di-O-methylquercetin, exhibited favorable ADMET profiles, low cytotoxicity, and stable binding to sEH during molecular dynamics simulation (equilibrated RMSD ∼0.8 nm with fluctuations <0.1 nm; MM-PBSA free energies: –117.49±14.01 and –71.47±13.87 kJ/mol). Both compounds reduced NO and TNF-α levels in a dose-dependent manner in LPS-induced RAW264.7 macrophages. These findings identify metabolites of Oldenlandia diffusa as computationally predicted sEH inhibitors with confirmed anti-inflammatory activity, warranting further direct enzymatic validation. Conclusion Syringaresinol and 3,7-Di-O-methylquercetin from Oldenlandia diffusa are predicted sEH inhibitors with validated anti-inflammatory effects, supporting their further exploration as natural leads for therapeutic applications in inflammation.