Design, synthesis, molecular docking and molecular dynamics simulations of pyrazolo[3,4-d]pyrimidine scaffolds as selective COX-2 and CDK2 kinase inhibitors with dual anti-inflammatory and anti-proliferative activities
A new series of pyrazolo[3,4-d]pyrimidine scaffolds, namely, 4a–l, were designed, synthesized, and tested for anti-inflammatory and anti-proliferative activities. Compounds 4f and 4g displayed advanced COX-2 selectivity (SI = 4.95 and 16.22) in comparison to celecoxib (SI = 7.61). In addition, all compounds were evaluated for their cytotoxic activity against three cancer cells using a reference drug (staurosporine). Compounds 4f and 4g showed the most potent anti-proliferative activity against MCF-7 (IC50 = 0.259 and 0.292 µM, respectively), MDA-MB-231 (IC50 = 0.258 and 0.327 µM, respectively), and Caco-2 (IC50 = 0.228 and 0.178 µM, respectively) cell lines and were more potent than staurosporine (IC50 = 0.401, 0.411, and 0.224 µM, respectively). Moreover, both derivatives were less toxic with comparable selectivity to the normal breast cell line (MCF-10) (SI values of 1.13–4.43) compared to staurosporine (SI values of 1.10–2.02). Further assessment of the CDK2 inhibitory activity of the most cytotoxic candidates 4f and 4g revealed their higher potency (IC50 = 0.324 and 0.215 µM, respectively) than that of the reference drug ribociclib (IC50 = 0.496 µM). Additionally, compound 4g revealed cell cycle arrest at the G2/M phase in Caco-2 cells, resulting in cell apoptosis. Furthermore, molecular docking and dynamics simulations revealed that derivatives 4f and 4g form stable and tighter complexes with the CDK2 kinase domain, as confirmed by their lower RMSD values, superior interaction stability, and stronger hinge and catalytic site binding. Finally, ADME profiling demonstrated that compound 4f satisfies all major drug-likeness criteria without any violations, while compound 4g fully adheres to the Veber's, Lipinski's, Egan's, and Ghose's drug-likeness guidelines. Consequently, these compounds represent promising starting candidates for the development of future CDK2 kinase and COX-2 inhibitors.