A highly efficient one-pot approach for the synthesis of novel 5-amino-4-Cyano-Pyrazoles with promising anti-inflammatory and broad-spectrum antimicrobial activity.

A green and efficient one-pot synthetic method has been developed to prepare novel 5-amino-3-(aryl)-1-(aryl)-1H-pyrazole-4-carbonitrile analogues, employing ammonium acetate as a mild, green catalyst. The proposed methodology provides a rapid, operationally simple, and sustainable approach for the construction of biologically relevant heterocyclic scaffolds with good yields. Confirmation of the synthesized compounds was achieved using IR, 1H NMR, 13C NMR, and HRMS. All synthesized derivatives were subjected to in vitro biological assessment for their anti-inflammatory, antitubercular, antibacterial, and antifungal activities. Several compounds demonstrated promising pharmacological profiles. In particular, compounds 4d, and 4f exhibited significant COX-2 inhibitory activity, with IC50 value of 77.62 and 65.95 μg/mL. Antitubercular screening revealed compounds 4d and 4e showed enhanced potency, displaying sensitivity at the MIC value of 3.12 μg/mL, compared to the standard reference drug pyrazinamide. Furthermore, the synthesized derivatives exhibited notable antibacterial and antifungal activities comparable to reference drugs, indicating their broad-spectrum antimicrobial potential. Computational studies, including molecular docking, MD simulations, and MM-GBSA, provided further support for the observed COX-2 inhibition and indicated favorable pharmacokinetic profiles. These findings highlight the synthetic efficiency of the developed one-pot protocol and the promising biological potential of the synthesized 5-Amino-4-cyanopyrazole derivatives. Further studies involving structural optimisation, in vivo assessment, and mechanistic investigations are warranted to advance these compounds toward pharmaceutical development.