Comparative Study of Conventional, Grinding, and Microwave‐Assisted Synthesis of Aminopyrazolones and Diaminopyrazoles: Exploring the Antitumor Activity, Dual CDK‐2/CA IX Inhibition Potential, and Apoptosis Induction

Abstract

ABSTRACT Embracing drug design approaches including ring variation, substituent variation, and bioisosteric modifications, aminopyrazolones 2–7 and diaminopyrazoles 9–14 were synthesized as dual potent CDK‐2 and CA IX inhibitors for the first time. The eco‐friendly preparation of the target analogs was performed by three procedures: conventional, grinding, and microwave‐assisted methods. The synthesized congeners were estimated for their antitumor effect against breast MCF‐7, hepatocellular HepG2, and colon HCT‐116 cells where the aminopyrazolones 4 and 7 presented significant cytotoxicity against the examined carcinomas. Compound 4, bearing a dinitrophenyl ring at N ‐2 of aminopyrazolone scaffold, exhibited the greatest cytotoxicity and selectivity toward the tested cell lines. Hence, compounds 4 and 7 were selected for consecutive biological assays to determine their mode of action. The findings proposed that 4 and 7 may exert their antiproliferative activity via interaction with CDK‐2 and CA IX receptors. Entity 4 exhibited promising dual inhibition of CDK‐2 and CA IX with IC 50 at the micromolar level, which exceeded that of Roscovitine by three times and nearly half that of acetazolamide. Additionally, the superior derivative 4 stimulated MCF‐7 cycle arrest at S phase through apoptotic induction which is supported by the upregulation of Bax and Caspase‐8 and the downregulation of Bcl‐2 and Cyclin E. The in silico studies showed acceptable predicted ADME and physicochemical properties together with the strong interaction between the superior compounds and both CDK‐2 and CA IX binding sites inspiring such hybrids as potential lead dual inhibitors.