Design, synthesis, molecular docking, and biological evaluation of chalcones bearing benzotriazole analogues as dual anti-inflammatory and anticancer agents.

Current NSAIDs and chemotherapeutics suffer from significant toxicities and acquired resistance, creating a pressing need for safer agents. Benzotriazole is a "privileged scaffold" for designing novel therapeutics. This study reports the rational design of novel benzotriazole-chalcone analogues as dual-target inhibitors of cyclooxygenase-2 (COX-2) and aromatase, leveraging their established molecular crosstalk in carcinogenesis. Eight novel benzotriazole-chalcone analogues (PM-1 to PM-8) were synthesized and characterized by FTIR, 1H NMR, 13C NMR, and LCMS. A multi-faceted evaluation included in silico ADMET screening (SwissADME/ProTox-III) and molecular docking (Glide XP) against COX-2 (4COX) and aromatase (3EQM). Analogues were then experimentally validated via in vivo carrageenan-induced paw edema assays and ex vivo cytotoxicity screening against the MCF-7 cell line (SRB assay). All analogues demonstrated favorable in silico drug-likeness and high predicted GI absorption. Molecular docking revealed potent COX-2 binding: PM-6 (-10.519 kcal/mol) and (2E)-1-(1H-benzotriazol-1-yl)-3-(4-hydroxy-3-methoxyphenyl)-prop-2-en-1-one (PM-4) (-10.153 kcal/mol) exhibited stronger affinity than Diclofenac (-8.135 kcal/mol). In vivo, PM-4 and (2E)-1-(1H-benzotriazol-1-yl)-3-(3-hydroxy-4-methoxyphenyl)-prop-2-en-1-one (PM-6) produced significant (p < 0.001) paw edema inhibition. Ex vivo, (2E)-1-(1H-benzotriazol-1-yl)-3-(3-hydroxy-4-methoxyphenyl)-prop-2-en-1-one (PM-6) was the most potent, exhibiting 98.2% inhibition of MCF-7 cell growth at 80 µg/mL. This study identifies (2E)-1-(1H-benzotriazol-1-yl)-3-(4-hydroxy-3-methoxyphenyl)-prop-2-en-1-one (PM-4) and (2E)-1-(1H-benzotriazol-1-yl)-3-(3-hydroxy-4-methoxyphenyl)-prop-2-en-1-one (PM-6) as highly promising, dual-action lead compounds. The strong correlation between their potent in silico binding and experimentally-validated biological activities, combined with favorable ADMET profiles, establishes them as strong candidates for further preclinical development.