Acalypha wilkesiana Hoffmanni is a medicinal plant known to contain several bioactive secondary metabolites with reported therapeutic potential. This study investigated the antihypercholesterolemic activity and structural characterization of compounds isolated from the hexane leaf extract of A wilkesiana. Hypercholesterolemic rats exhibited elevated total cholesterol and low-density lipoprotein (LDL) levels compared to treated groups. Treatment with the isolated compound significantly reduced total cholesterol and LDL levels P 0.05 relative to untreated hypercholesterolemic rats. In addition, antioxidant enzyme activities, including glutathione peroxidase (GPx), superoxide dismutase (SOD), and catalase (CAT), were significantly increased, while malondialdehyde (MDA) levels were reduced P 0.05 Spectroscopic analysis revealed characteristic infrared absorption peaks at 1019,3, 1101.3, 1116.47, 1248.74, 1462.44, 1721.48, and 2859.17 cm. Mass spectrometry showed a molecular ion peak at m/z 406 (M + 2) corresponding to a molecular formula of C25H4204. Based on spectral data and comparison with literature, the compound was identified as ursodeoxycholic acid methyl ester. The isolated compound from A wilkesiana demonstrated significant antihypercholesterolemic and antioxidant effects, suggesting its potential as a candidate for further drug development.