3‐Substituted Triazolone–Benzoate Hybrids: Synthesis, Enzyme Inhibition, Anticancer Activity and ADMET Evaluation by Molecular Docking and Dynamics Studies

In this study, eight 3-substitued -4-amino-4,5-dihydro-1H-1,2,4-triazol-5-one compounds were synthesized. The reactions of these compounds with 2-ethoxy-4-formylphenyl benzoate, which was synthesized by the reaction of 3-ethoxy-4-hydroxy benzaldehyde with benzoyl chloride by using triethylamine, were investigated. Eight novel 2-ethoxy-4-(((3-substitued-5-oxo-1,5-dihydro-4H-1,2,4-triazol-4-yl)imino)methyl)phenyl benzoate compounds were obtained in order to identify the new synthesized compounds by spestroscopic methods including IR, ¹H-NMR and ¹³C-NMR used. Concentration ranges of 25, 50, 120, 200, and 400 µg/mL were applied to determine the compund's anti-cancer properties. The human ovarian cancer cell line (OVCAR-3) compared with human umbilical vein endothelial cells (HUVEC) and significant results were obtained against the OVCAR-3 cell line. The enzyme inhibitory effect of compound on glutathione S-transferase (GST), acetylcholinesterase (AChE) enzymes were studied. Ki and IC₅₀ values were found in the range of 1.1143 ± 0.2402 µM-7.9100 ± 1.9107 µM and 1.840 µM-4.149 µM for AChE; 2.0733 ± 0.8199 µM-8.2120 ± 1.5720 µM and 1.320 µM-3.223 µM for GST. In silico ADMET and drug-likeness analyses confirmed favorable pharmacokinetic and drug-like profiles for the compounds. Additionally, molecular docking and 100 ns molecular dynamics (MD) simulations revealed stable binding modes and dynamic stability of the most active compounds with the target enzymes.