Computational and experimental insights into the anticancer activity of benzylidene amino benzoate derivatives: A study based on docking, DFT, and in vitro assays

Abstract

Series of benzylidene amino benzoate derivatives were synthesized under mild conditions, chemical structure of the synthesized compounds was confirmed through different spectroscopic analysis. Anticancer activity was evaluated against four cancer cell lines (PC3, A549, A2780, and MDA-MB-231). The results showed potent cytotoxic effects with notable selectivity. Compounds 5, 6, 9, and 10 displayed significant activity, with IC₅₀ values ranging from 0.11 µM to 1.37 µM. Molecular docking studies indicated favourable binding affinities for compound 8 toward different tyrosine kinases receptors, with the highest affinity for binding to VEGFR2. This result was consistent with 76.25 % in vitro inhibition activity against VEGFR2. Docking simulations revealed that hydrogen bonds could form between compound 8 and the residues Asp 1046 and Lys 868, while electrostatic interactions were observed with residues His 1026 and Glu 885, at the ATP-binding pocket. The compounds complied with Lipinski's Rule of Five, supporting their suitability for further development as orally active drugs. Non-substrate behaviour toward P-glycoprotein was observed for the synthesized compounds, implying they were unlikely to be expelled by this transporter. DFT calculations highlighted important reactive regions within the target compound, particularly around the nitro and ester groups, which participated in hydrogen bonding interactions. Furthermore, FMO suggested versatility in the chemical reactivity. Enhanced inhibition of VEGFR2 by compounds like 5 and 8 was predicted with lower chemical potential and higher electrophilicity indices. Based on these findings, imine compounds should be prioritized for the synthesis of more derivatives and for in vivo studies to enhance their therapeutic efficacy as anticancer agents.