Dual COX-2 and EGFR inhibition by pyrazole-ar-turmerone hybrids suppresses colorectal cancer cell proliferation.

Colorectal cancer (CRC) progression involves a coordinated interaction between COX-2-mediated inflammation and EGFR-driven proliferation. Current monotherapies often fail due to incomplete pathway suppression and resistance development, highlighting the need for multi-targeted strategies. This study aimed to design and synthesize novel pyrazole-ar-turmerone hybrids capable of simultaneously inhibiting COX-2 and EGFR, thereby achieving enhanced anti-proliferative efficacy in inflammation-associated colorectal cancer. Two hybrid molecules (compounds 1 and 2) were synthesized and characterized. Their dual-target potential was evaluated in silico using network pharmacology and molecular docking against COX-2 and EGFR crystal structures. In vitro assays in IL-1β-stimulated HT-29 colorectal cancer cells were used to assess anti-proliferative effects by MTT, clonogenic, and CFSE flow cytometry analyses. Mechanistic studies were performed through Western blotting and PGE2 ELISA/rescue experiments to examine inhibition of COX-2 activity and EGFR-ERK1/2 signaling. Both compounds showed high COX-2 selectivity (SI = 14.38 and 23.57) and potent COX-2 inhibition (IC50 = 0.63 and 1.04 μM), together with EGFR kinase inhibition (IC50 = 37.56 and 57.56 μM). Both hybrids exhibited low cytotoxicity yet significantly suppressed IL-1β-induced HT-29 cell proliferation, with GI50 values of 5.85 μM (compound 1) and 9.57 μM (compound 2). Mechanistic analysis confirmed reduced PGE2 production, inhibition of EGFR-ERK1/2 activation, and downregulation of cyclin D1 and PCNA expression. The pyrazole-ar-turmerone hybrids function as potent dual COX-2/EGFR inhibitors exhibiting selective anti-proliferative activity in inflammation-driven CRC. These compounds represent promising leads for the development of next-generation dual-target therapeutics against colorectal cancer therapy.