Synthesis, molecular docking, and in vitro cytotoxicity of novel N-(α-cyanocinnamoyl)hydralazine derivatives

Background and purpose: Treatment resistance in many cancers is a major concern, requiring the development of new anticancer agents to overcome it. Hence, in the present study, a novel series of (2 E )-2-cyano-3-substituted phenyl- N '-(phthalazin-1-yl)prop-2-enehydrazide derivatives ( 4a-h ) were synthesized and tested against different cancer cell lines to explore their potent anticancer activity. Experimental approach: The structures of the synthesised derivatives were confirmed by FTIR, 1 H NMR, 13 C NMR, and mass spectrometry. Compounds were assessed for in vitro cytotoxicity using molecular docking, the MTT assay, an in vitro EGFR inhibition assay, molecular dynamics simulations, and ADMET predictions. Findings/Results: Molecular docking studies revealed that compound 4f exhibited favorable binding affinity towards EGFR (-7.79025 Kcal/mol) and hERα (-8.64251 Kcal/mol), while compound 4b showed good docking interactions with the Bcl-2 protein (-5.08782 Kcal/mol), comparable to the respective standards. In vitro cytotoxicity assays showed that compound 4f elicited the highest cytotoxicity against A549 (IC 50 = 12.5 ± 1.3 μM) and MCF-7 cell lines (IC 50 = 18.0 ± 1.5 μM), along with significant EGFR inhibition (IC 50 = 7.6 ± 0.9 μM). compound 4b showed appreciable cytotoxicity against PC-3 cell lines (IC 50 = 9.0 ± 0.9 μM). Molecular dynamics simulations further confirmed the stability of the 4f- EGFR complex. ADMET studies indicated that all the synthesized compounds exhibited acceptable pharmacokinetic profiles, with most descriptors falling within the recommended range for known drugs. Conclusion and implications: The present study identified compound 4f as a lead scaffold for further research to validate and advance its therapeutic applicability.