Study on the anti-glycation active constituents of Withania somnifera based on molecular docking and experimental verification

Abstract Withania somnifera (L.) Dunal, commonly known as Ashwagandha, has been demonstrated in prior studies to delay skin glycation damage, yet the specific active constituents underlying this pharmacological effect remain unclear. Bioactive compounds identified from WS ( W. somnifera root extract) by LC-MS were subjected to molecular docking against RAGE, a key protein target associated with glycation-related skin damage. Based on binding energy levels, compounds showing the lowest binding energy were selected and subsequently evaluated in a MGO-induced glycation-damaged skin fibroblast cell model, with RAGE receptor protein expression examined by Western blotting as a preliminary validation of the docking results. In parallel, ADMET prediction using the pkCSM platform indicated generally favorable theoretical bioavailability and low toxicity, while limited skin permeability, potential hERG II inhibition, AMES mutagenicity and Lipinski violations were also noted as safety concerns. The results showed that seven active compounds, including Withanolide B, lowered MGO-elevated RAGE expression to some extent, resembling the effects of FPS_ZM1 and WS. It should be emphasized that these findings remain preliminary, being based on computational screening, ADMET prediction and a single biomarker, and thus warrant further validation. Nevertheless, they provide a valuable basis for the continued utilization of W. somnifera resources in the development of anti-glycation agents.