Exploring Withania somnifera L. as a Natural Remedy for Dysmenorrhea treatment via COX-2 Inhibition

AbstractThe present work was aimed to evaluate the anti-inflammatory effect of bioactive compounds ofWithaniaSomnifera L. against cyclooxygenase-2 (COX-2) targeting dysmenorrhea using an in-silico molecular docking approach. Due to the limitations and adverse effects associated with currently available anti-inflammatory drugs, the identification of safer natural COX-2 inhibitors has gained significant attention. A phytochemical library comprising 16 bioactive compounds of Withaniasomnifera L.,commonly known asAshwagandha and belonging to the Solanaceae family,previously reported for their anti-inflammatory,antidepressant, anticancer and antianxiety activities, was selected for the study.The three-dimensional crystalstructure of the target protein cyclooxygenase-2 (COX-2) was retrieved from the Protein Data Bank (PDB)(PDBID: 1CX2), while the chemical structures of phytochemicals were obtained from the PubChem database. Activesite prediction and binding pocket analysis of the target protein were carried out using CASTp (Computed Atlasof Surface Topography of Proteins). Molecular docking and virtual screening studies were performed usingPyRx, with Celecoxib used as the reference inhibitor. Furthermore, protein–ligand interaction analysis wasvisualized using BIOVIA Discovery Studio Visualizer. The docking results revealed that 13 phytochemicalsexhibited stronger binding affinities than the reference inhibitor Celecoxib (-9.2 kcal/mol) against the COX-2active site. Among these, representative top-ranked compounds including Withaferin A (-11.5 kcal/mol),Somniferanolide(-11.4 kcal/mol), Withaoxylactone (-11.3 kcal/mol) and Withanolide A (-11.1 kcal/mol) werehighlighted in the present study due to their comparatively superior binding affinities and significant interactionswith key active-site residues of the COX-2 enzyme. These compounds formed multiple hydrogen bonds andother stabilizing interactions, indicating strong inhibitory potential. Overall, the findings suggest thatphytochemicals from Withaniasomnifera L.may serve as promising natural anti-inflammatory agents for themanagement of dysmenorrhea. However, further in-vitro and in-vivo studies are required to validate theirtherapeutic efficacy and safety.