Background: The synthesis of novel heterocyclic scaffolds with amide functionality is a key research
area due to their plethora of medicinal applications. The 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±1.01μM and 4.05±0.62μM against HeLa cell lines. The flow-cytometry analysis demonstrated
that the compound 5eb arrested the sub-G1 phase of the cell cycle and induced apoptosis. Furthermore, the compound
5eb triggered the collapse of mitochondrial membrane potential (ΔѰm), which was assessed by JC-1
staining and also induced the generation of Reactive Oxygen Species. An increase in the expression of proapoptotic
proteins such as Bax, p53, cleaved PARP and cleaved caspase-3 by 5eb confirmed the activation of
the 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.