Molecular Targets and Pharmacological Mechanisms of Anti-Hepatocellular Carcinoma Activity of Compounds Isolated from Bridelia ferruginea in Nigerian-Context Liver Cancer Research

Background: Hepatocellular carcinoma (HCC) is one of the most prevalent and lethal cancers in Nigeria, driven by high rates of chronic hepatitis B and aflatoxin exposure. Current pharmacological options for advanced HCC are expensive and inaccessible to most Nigerian patients. Bridelia ferruginea, a plant used in South West and South South Nigerian ethnomedicine for liver conditions, contains phytochemicals with reported cytotoxic properties. This study investigates the molecular mechanisms of anti-HCC activity of isolated B. ferruginea compounds. Methods: Bioguided fractionation of B. ferruginea stem bark identified three candidate compounds characterised by NMR and MS. In vitro cytotoxicity was assessed against HepG2 and Huh-7 HCC cell lines. Mechanistic investigations employed flow cytometry for apoptosis and cell cycle analysis, Western blotting for apoptosis pathway proteins (Bcl-2, Bax, caspase-3/9), and molecular docking against EGFR and CDK2 using AutoDock Vina. In vivo antitumour activity was assessed in a xenograft mouse model. Results: Compound BF-2 (tentatively identified as a pentacyclic triterpenoid) demonstrated selective cytotoxicity (IC50 4.8 uM in HepG2). Apoptosis induction via the intrinsic mitochondrial pathway was confirmed. Molecular docking revealed strong binding affinity to CDK2. In vivo tumour volume reduction was 54 percent at 50 mg/kg. Conclusion: BF-2 from B. ferruginea is a promising anti-HCC lead compound targeting CDK2 and inducing intrinsic apoptosis. These findings warrant pre-clinical development and IND application. Keywords: hepatocellular carcinoma, Bridelia ferruginea, apoptosis, CDK2, natural products