1, 2, 4-Triazole-benzoyl aza glycinamide hybrids: design, synthesis, biological evaluation, and computational investigations on anticancer targets

The 1, 2, 4-triazole ring and the azaglycine are two pharmacologically significant structural motifs associated with cytotoxic activity. In particular, the aza glycine scaffold exhibits enhanced affinity towards cathepsin B enzyme, an established molecular target for breast cancer. Accordingly, twelve 1,2,4-triazole-benzoyl azaglycinamide hybrids (3a-3l) bearing diverse substituents on the phenyl ring of the benzoyl moiety were synthesized and evaluated for anticancer activity using the MTT assay. The synthesized compounds were further screened for antibacterial (Staphylococcus aureus, Bacillus subtilis, Escherichia coli, and Proteus vulgaris) and antioxidant potential. Among them, compound 3l, containing a 3,5-dinitro substituent, demonstrated notable cytotoxicity against the MCF-7 cell line, with an IC50 value of 27.32 µM, compared to cisplatin (12.06 µM). Molecular docking studies indicate that the compounds 3f (4-OH), 3c (4-NH2), and 3 g (2-OH, 3,5-(NO2)2) formed more hydrogen bonds and higher affinity for cathepsin B (PDB ID: 8B4T), while 3g and 3d (3,5-OCH3)2 showed favourable binding with EGFR (PDB ID:1M17). Prime MM-GBSA analysis and ligand strain energy calculations further suggested moderate to high binding affinity of the synthesized hybrids towards cathepsin B. However, computational predictions did not consistently correlate with the experimental cytotoxicity for the most active compound 3l. Notably, compound 3l also displayed potent antibacterial activity against the tested organisms. Overall, these findings indicate that the rational structural modifications of these hybrids might enhance their biological potential.