Lenvatinib, a first-line multi-kinase inhibitor targeting VEGFR-2 tyrosine kinase, demonstrates significant therapeutic potential in hepatocellular carcinoma (HCC) treatment. Guided by structure-based rational design targeting, the critical binding domains of VEGFR-2 and lenvatinib, a novel series of aryl-substituted lenvatinib derivatives were synthesized via palladium-catalyzed Suzuki cross-coupling reactions. Systematic evaluation revealed superior anti-proliferative effects of these derivatives against HepG2 and Hep3B human HCC cell lines compared to the parent compound. Particularly, the 3-isopropyl aryl derivative 3i exhibited threefold enhanced potency (IC50 = 2.4 ± 0.28 μM) over lenvatinib. Mechanistic investigations demonstrated that compound 3i significantly promoted HCC cell apoptosis through caspase-dependent pathways. The IC50 values of compounds 3i against VEGFR-2 kinases (IC50 = 22.2 ± 0.52 nM) is showed activity comparable to that of lenvatinib (IC50 = 16.4 ± 1.90 nM). Molecular docking and 100 ns molecular dynamics simulations further elucidated that the 3i achieved enhanced binding affinity through optimized hydrophobic contacts within the ATP-binding pocket of VEGFR-2. This structure-activity relationship study provides valuable insights for developing next-generation VEGFR-2 inhibitors, with compound 3i emerging as a promising drug candidate warranting further preclinical evaluation.
