Synthesis and evaluation of Urolithins derivatives as anticancer agents for hepatocellular carcinoma: In vitro, molecular docking, and dynamics simulations

In this study, 23 novel urolithins derivatives were designed and synthesized, in order to discover promising anti-hepatocellular carcinoma candidates. The antiproliferative activities of 5a-i, 7a-g and 11a-h against three different kinds of human hepatocellular carcinomar cell lines (HepG2, Hep3B and Huh-7) were determined. Among the synthesized compounds, compound 11e exhibited the most potent anti-proliferative activity against HepG2 cell lines and Hep3B cell lines, with IC₅₀ values of 4.55 ± 0.76 μM and 4.95 ± 0.97 μM, respectively, and was selected for further study in HepG2 cells. Subsequently, in vitro studies revealed that compound 11e significantly inhibited HCC cell migration, induced cell cycle arrest at both the G2/M phases, and promoted Apoptosis in a dose-dependent manner. Moreover, integrated network pharmacology and molecular docking analyses identified several key targets, including EGFR, Akt1, MAPK1, and CASP3, suggesting a multi-target mechanism involving regulation of the PI3K/Akt and MAPK signaling pathways. Subsequent molecular dynamics simulations confirmed the stability of the corresponding protein-ligand complexes, as indicated by low RMSD and RMSF values and consistent hydrogen bond interactions. Western blot analysis confirmed that compound 11e effectively suppressed the phosphorylation of Akt (Ser473) and ERK1/2 (Thr202/Tyr204), further validating the inhibition of the PI3K/Akt and MAPK/ERK signaling axes. Overall, compound 11e demonstrated promising anti-tumor efficacy and was identified as an early-stage lead compound for further development as an anti-HCC agent.