Synthesis, spectroscopic characterization, and multiscale computational ınvestigation of a novel 1,2,4-triazole derivative targeting AChE, BChE, and COX enzymes

Abstract

In this study, a new 1,2,4-triazole derivative, 4-(((3-ethyl-5-oxo-1,5-dihydro-4H-1,2,4-triazol-4-yl)imino)methyl)-2-methoxyphenyl benzoate (MPB), was synthesized and structurally confirmed using FTIR, UV–Vis, and ¹H and ¹³C-NMR spectroscopy. The electronic properties were examined at the B3LYP/6-311G(d,p) level through frontier molecular orbital (FMO) analysis, which revealed a HOMO–LUMO gap of 4.36 eV, indicating significant kinetic stability. Nonlinear optical (NLO) analyses revealed a first-order hyperpolarizability (β_tot) of 2.09 × 10⁻³⁰ esu, approximately 11 times greater than that of urea, suggesting strong NLO potential. Global reactivity descriptors showed a high ionization potential (6.18 eV), electron affinity (1.82 eV), and electrophilicity index (8.25 eV), indicating MPB’s stability and moderate reactivity. MEP, ELF, and LOL analyses highlighted electrophilic and nucleophilic regions of MPB. Molecular docking studies demonstrated that MPB binds effectively to acetylcholinesterase (AChE), butyrylcholinesterase (BChE), and cyclooxygenases (COX-1 and COX-2), with interactions including π–π stacking (e.g., with Trp86, Phe297, and Tyr385) and multiple hydrogen bonds. Induced Fit Docking revealed conformational adaptability within the active sites. Molecular dynamics (MD) simulations over 200 ns confirmed the structural stability of MPB–protein complexes, with RMSD values around 1.5–2.0 Å. Markov State Model analyses (PCA, TICA-FES) further revealed distinct energy landscapes and conformational constraints, particularly for AChE and BChE, where MPB showed superior stability compared to reference inhibitors. These findings underscore MPB’s potential as a multifunctional pharmacophore with notable enzyme-inhibitory capabilities.