Comprehensive Assessment of Hyperoside: Cytotoxic, Anti-cancer, Genoprotective, Radical Scavenging and Computational Toxicity Properties through <i>In Vitro</i> and <i>In Silico</i> Approaches

Hyperoside (HYP), a plant-derived flavonoid, was evaluated for its cytotoxic, anti-cancer, genotoxic, antigenotoxic, and antioxidant properties, and potential mechanisms of action using an integrated in vitro and in silico approach. Cytotoxicity and anti-cancer activities were assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay in CCD18-Co healthy colon epithelial and DLD-1 colon cancer cells. At the same time, genotoxicity and antigenotoxicity potentials were examined in human lymphocytes using COMET assay. Antioxidant activity was determined by 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay, and molecular docking was performed with key regulatory proteins (p53, ATM, 7O7B, and Keap1), and in silico toxicity profiling was conducted using the Percepta platform. HYP selectively reduced the viability of colon cancer cells (43.43% - 70.24%) without inducing cytotoxicity in healthy colon epithelial cells (cell viability percentage ranging from 82.25% - 111.88%) after 24 and 48 h of exposure. HYP did not significantly increase DNA damage at 7.81–62.5 μg/mL; however, it exhibited antigenotoxic effects at all concentrations, significantly reducing H2O2-induced DNA damage. HYP also showed significant antioxidant activity, with DPPH inhibition ranging from 53.09% to 78.64%. Docking analyses revealed strong binding affinities for ATM and Keap1, supporting its potential role in modulating oxidative stress and DNA damage response pathways. Percepta-based toxicity profiling predicted low to moderate acute systemic toxicity, limited oral bioavailability, and no major cardiotoxic or CYP-mediated safety liabilities; notably, predicted mutagenicity alerts were not corroborated by in vitro DNA damage assessments. Overall, these findings suggest that HYP exhibits selective anticancer and antigenotoxic properties, potentially mediated by its antioxidant activity.