Synthesis of Substituted Cinnamido Linked Quinazolinone Congeners as Potential Anticancer Agents via Mitochondrial Dependent Intrinsic Apoptotic Pathway

Background: The synthesis of novel heterocyclic scaffolds with amide functionality is a key research area due to their plethora of medicinal applications. Present study aims to explore the synthesis of new cinnamido linked quinazolinone congeners and their potential as anticancer agents. Methods: Cytotoxicity evaluation, Cell cycle analysis, JC-1 staining, ROS, Annexin V assays, AO/EB, DAPI nuclear staining, Colony forming assay and Western blot analysis. Results: Among the synthesized compounds 5eb and 5fc have shown promising cytotoxic activity with an IC50 value of 3.89 +/- 1.01 mu M and 4.05 +/- 0.62 mu M against HeLa cell lines. Flow-cytometry analysis demonstrated that the compound 5eb arrest the sub-G1 phase of cell cycle and induce apoptosis. Furthermore the compound 5eb trigger the collapse of mitochondrial membrane potential (Delta Psi m), which was assessed by JC-1 staining and also induce the generation of Reactive Oxygen Species. Increase in the expression of pro-apoptotic proteins such as Bax, p53, cleaved PARP and cleaved caspase-3 by 5eb confirms the activation of mitochondrial dependent intrinsic apoptosis pathway. Conclusion: Our results suggest that compound 5eb and 5fc of cinnamido linked quinazolinone derivatives could serve as potential leads in the development of novel chemotherapeutic agents.