In Vitro Cholinesterase Inhibition and Computational Investigation of some 1,2,3-triazole Derivatives as Dual AChE/BuChE Inhibitors

In vitro anticholinesterase activities of a series of 1,2,3-triazole derivatives, 9(a-e) and 10(f-l), were evaluated against electric eel acetylcholinesterase (eeAChE) and equine serum butyrylcholinesterase (eqBuChE), using donepezil as a reference inhibitor. Enzyme inhibitory activities were characterized by IC50, inhibition constants (Ki), and inhibition mechanisms. All target compounds exhibited potent inhibitory activity against both enzymes. 9e demonstrated the highest activity against eeAChE (Ki = 7.69 ± 1.24 nM), outperforming donepezil (Ki = 25.28 ± 5.79 nM). 9b showed the strongest inhibition against eqBuChE (Ki = 3.36 ± 0.67 nM), whereas donepezil displayed comparatively weak activity (Ki = 43.48 ± 10.36 nM). To further elucidate ligand–enzyme interactions, molecular docking, molecular dynamics simulation (MDS), MM/GBSA, and structure–activity relationship studies were performed. Docking results indicated that all target compounds exhibited higher binding affinities for both the catalytic active site (CAS) and peripheral anionic site (PAS) of AChE than for BuChE. MDS and MM/GBSA analyses confirmed the stability of the selected complexes, without significant fluctuations over 50 ns, and showed that while 9e was selective for hAChE-CAS, 9b bound similarly to both hBuChE sites. Taken together, all target compounds, particularly 9e and 9b, may serve as novel dual AChE/BuChE inhibitor candidates. © 2026 Wiley-VCH GmbH.