Extraction and Pharmacological Validation of Ursolic Acid from Hedyotis diffusa: A Molecular Docking Approach against Inflammation and Colon Cancer

In traditional Chinese and Indian medical systems, Hedyotis diffusa is a well-known medicinal plant that is renowned for its wide range of pharmacological properties. The extraction, identification and pharmacological confirmation of ursolic acid a significant bioactive triterpenoid from H. diffusa are the main objectives of this investigation. The ethanolic extract was subjected to Soxhlet extraction, followed by column chromatography for the isolation of ursolic acid, which was subsequently confirmed by GC-MS analysis. A major peak at retention time 38.25 min and a molecular ion peak at m/z 456 confirmed its identity. The anti-inflammatory properties of ursolic acid were assessed using protein denaturation and membrane stabilization tests, showing notable suppression in a dose-dependent manner that was on par with that of regular diclofenac. Moreover, the MTT assay was used to evaluate its anticancer effectiveness against colon cancer cell lines (HT-29), where ursolic acid exhibited distinguished cytotoxicity with an IC50 value suggestive of strong anti-proliferative effects. To gain mechanistic insights, molecular docking studies were performed using AutoDock, targeting COX-2 (PDB ID: 1CX2) for inflammation and TNIK (PDB ID: 6GUE) for colon cancer. Ursolic acid displayed high binding affinities and stable interactions with both targets, indicating its potential dual inhibitory action. These results support the therapeutic relevance of ursolic acid from H. diffusa as a promising natural compound for managing inflammation and colon cancer.