Hepatocellular carcinoma (HCC) remains fatal malignancy commonly nurtured by the aberrant Fibroblast Growth Factor Receptor 4 (FGFR4) signaling pathway. The existing clinical inhibitors acting on this pathway are challenged by resistance, off-target toxicity, and the need to be replaced with new, safe chemical scaffolds. This study used a "phenotype-first" computational strategy to identify selective FGFR4 inhibitors from a library of 216 bioactive phytochemicals, which were subsequently experimentally validated. The library was screened against the FGFR4 kinase domain using molecular docking, and the stability of the top hits was assessed using 100 ns molecular dynamics (MD) simulations and MM/GBSA free energy calculations. In vitro activity was estimated using MTT, wound healing, and clonogenic assays in HEP3B cells. 7-Methoxyflavone emerged as the lead candidate, with a binding affinity of -9.6 kcal/mol, surpassing the clinical control Fisogatinib (-8.6 kcal/mol). MD simulations confirmed a stable interaction (RMSD ≈ 0.20 nm) and a robust binding free energy of -31.6 kcal/mol. In vitro assays demonstrated an IC50 of 51.50 μg/mL against HEP3B cells. Importantly, the compound showed minimal toxicity in normal HEK293 and VERO cells, with an improved Selectivity Index (SI) of >19. The anti-metastatic potential was confirmed by functional assays, in which 7-Methoxyflavone had a significant inhibitory effect on cell migration (∼15% wound closure vs. 100% in the control; p < 0.001) and colony formation. 7-Methoxyflavone is identified as a safe, selective lead hit targeting FGFR4. Whereas structural optimization is required to increase the potency. Its high selectivity offers a promising starting point for developing safer HCC therapeutics.
Thombare et al. (Sun,) studied this question.