Synthesis and CDK2 inhibition profiling of novel pyrazolyl-thiourea derivatives as potential anti-inflammatory agents

Background & objective: Cyclin-dependent kinases (CDKs), particularly CDK2, are essential regulators of cell cycle progression and have recently been implicated in inflammation and immune modulation. Inhibitors like Rescovitine promote neutrophil apoptosis, reduce cytokine release, and show anti-inflammatory. However, its short half-life and off-target effects limit its clinical utility. To synthesize and evaluate five novel thiourea derivatives of N-((4-cyano-1H-pyrazol-5-yl) carbamothioyl) benzamide that not only structurally resemble Rescovitine but also exhibit enhanced biological activity—particularly in inflammation control—through potent and selective inhibition of CDK2. This study aims to identify compounds with superior anti-inflammatory efficacy by targeting CDK2, a key regulator of neutrophil apoptosis and inflammatory resolution, while also offering improved pharmacokinetic properties. Methodology: The derivatives VIa–VIe were synthesized through a multi-step reaction sequence, and their CDK2 inhibitory activity was evaluated in vitro using the MTT assay on human cell lines. The IC₅₀ values were determined for each compound. Results: All compounds showed varying levels of CDK2 inhibition. VIa and VIe demonstrated the highest inhibitory activity with IC₅₀ values of 1.28 µM and 0.94 µM, respectively, comparable to Rescovitine (0.60 µM). VId and VId had moderate effects, while VIb was the least active. Fluorine—substitution in VIe enhanced activity, suggesting a key role of electronic substituents in CDK2 binding. Conclusion: VIa and VIe are biologically promising anti-inflammatory candidates, and their ability to inhibit CDK2 could make them useful in treating inflammatory and proliferative diseases, where CDK2 plays a critical pathological role. Further preclinical studies could confirm their therapeutic value. Abbreviations: CDK: Cyclin-dependent kinase, CDKI: CDK inhibitor, IL-10: Interleukin-10, MTT: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, TGF-β: Transforming growth factor beta, TLC: thin layer chromatography, C NMR: Computational nuclear magnetic resonance Keywords: Anti-inflammatory agents; CDK2 inhibition; Inflammation; Kinase inhibitors; Rescovitine; Thiourea derivatives Citation: Awad KT, AL-Salman HN, Najm MAA. Synthesis and CDK2 inhibition profiling of novel pyrazolyl-thiourea derivatives as potential anti-inflammatory agents. Anaesth. pain intensive care 2025;29(6):594-599; DOI: 10.35975/apic.v29i6.2855 Received: July 31, 2025; Revised: August 05, 2024; Accepted: August 05, 2025