Design, Synthesis, and Characterization of Novel Substituted 1,2,4‐Triazines: Cytotoxic Activity on HepG2 and HT‐29 Cell Lines, Enzyme Inhibition, and Molecular Docking Studies

ABSTRACT 1,2,4‐Triazines exhibit various pharmacological properties due to their strong biological activities, such as anticancer, anti‐inflammatory, antimicrobial, antiviral, and antioxidant, and also occupy an important place in the pharmaceutical field. The tested compounds were synthesized and characterized. 1,2,4‐Triazines were evaluated for their inhibitory and cytotoxicity effects on acetylcholinesterase (AChE), glutathione S‐transferase (GST), human liver cancer cell line (HepG2), and human colorectal adenocarcinoma cell line (HT‐29). 1,2,4‐Triazines ( 3a ‐ c , 5 , 6 , 10 ) were found to inhibit these enzymes with IC 50 values ranging from 1.41 to 4.28 μM for AChE and from 1.58 to 4.13 μM for GST. 1,2,4‐Triazine compounds were tested in range of 12.5−100 µM concentrations against HepG2 and HT‐29 cell lines. Among the 1,2,4‐triazines tested, 10 was the most effective substance on HepG2 cells, especially at high doses, while 3b was effective on HT‐29. Furthermore, molecular docking results indicated that the 1,2,4‐triazines showed strong binding and stability at the active site of enzymes and selected proteins. 1,2,4‐Triazines may serve as promising precursors for the design of potent enzyme inhibitors and anticancer agents.