Design, synthesis, computational evaluation, and in vitro cytotoxicity assessment of coumarin-3-carboxamide derivatives against HeLa cervical cancer cells

Introduction Coumarins are privileged scaffolds in medicinal chemistry, exhibiting broad spectra of biological activities; including anticancer, anti-inflammatory, antioxidant, and antimicrobial effects. In the present study, we synthesized a series of coumarin-3-carboxamide derivatives and assessed their potential as anticancer agents. Methods A series of N -R-2-oxo-2 H -chromene-3-carboxamide derivatives were synthesized by reacting 2-oxo-2 H -chromene-3-carbonyl chloride with the corresponding amines. In silico molecular docking studies were performed against human carbonic anhydrase (hCA) receptors IX and XII. Molecular dynamics simulations were performed for 100 ns to assess the stability of ligand-protein complexes. In vitro cytotoxicity was evaluated using MTT assay against HeLa (cervical cancer) and Vero (normal kidney) cell lines. Anticancer selectivity was determined using the selectivity index (SI). Results Molecular docking studies revealed strong binding affinities of the synthesized derivatives toward both hCA IX and XII. MD simulations showed compound 5 with lowest RMSD fluctuations in complex with hCA IX, and compound 4e with the highest stability toward hCA XII. Among the derivatives, compound 4 h exhibited most promising anticancer activity (IC 50 = 43.02 ± 1.36 µM in HeLa; 111.19 ± 1.74 µM in Vero) with a SI of 2.58 indicating selectivity to cancer cells. Standard drug doxorubicin showed superior potency with an SI of 5.78, but few coumarin derivatives demonstrated moderate selectivity. Conclusion Coumarin-3-carboxamide derivatives demonstrated computationally favourable binding affinity toward hCA IX/XII, with compound 4 h exhibiting highest in vitro activity and selectivity. The results can serve as a starting point to propose compound 4 h as a scaffold for further investigation as anticancer agent.