Background: Alzheimer’s disease (AD) is characterized by cholinergic dysfunction, making the inhibition of acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) critical for improving cholinergic neurotransmission. However, the development of effective dual inhibitors remains challenging. Objective: This study aims to synthesize and evaluate novel pyridazine-containing compounds as potential dual inhibitors of AChE and BuChE for AD treatment. Methods: Ten novel pyridazine-containing compounds were synthesized and characterized using IR, ¹H NMR, and ¹³C NMR. The inhibitory activities against AChE and BuChE were assessed in vitro, and pharmacokinetic properties were explored through in silico ADME studies. Molecular dynamics simulations were performed for the most active compound. Results: Compound 5 was the most potent inhibitor, with IC₅₀ values of 0.26 µM for AChE and 0.19 µM for BuChE, outperforming rivastigmine and tacrine, and showing competitive results with donepezil. Docking studies revealed a binding affinity of −10.21 kcal/mol to AChE and −13.84 kcal/mol to BuChE, with stable interactions confirmed by molecular dynamics simulations. In silico ADME studies identified favorable pharmacokinetic properties for compounds 5, 8, and 9, with Compound 5 showing the best activity. Conclusions: Compound 5 demonstrates strong potential as a dual cholinesterase inhibitor for Alzheimer’s disease, supported by both in vitro and in silico analyses. These findings provide a basis for further optimization and development of these novel inhibitors.

In this research, we successfully synthesized a distinctive group of iminothiazolidinone derivatives, using glucose isothiocyanate as the starting material. The structural elucidation of these newly created compounds was achieved through a combination of analytical techniques, including IR, 1H NMR, and 13C NMR. We then evaluated the inhibitory activity of these compounds against acetylcholinesterase (AChE) using the Ellman's method spectrophotometer, comparing their performance to standard drugs like donepezil, rivastigmine, and tacrine. Impressively, the majority of the tested compounds demonstrated inhibitory activity against AChE, with iminothiazolidinone derivative (5a) standing out as the most potent (IC50 = 0.209 μg/mL). It even surpassed the effectiveness of rivastigmine and tacrine, coming close to the potency of donepezil. Further investigation into the potential of these compounds as AChE inhibitors for Alzheimer's disease drug development involved docking simulations using Molecular Operating Environment (MOE). Derivatives 3,5-disubstituted-(2,3,4,6-tetra-o-acetyl-β-D-glucopyranosyl) imino) thiazolidin-4-ones (5a), (5f), (6c) and 3,5-disubstituted-(β-D-glucopyranosyl)imino)thiazolidin-4-one (6d) displayed promising docking scores in MOE simulations. In Silico ADMET experiments assessed their pharmacokinetic and toxicity studies, demonstrating strong binding affinity and favorable interactions with the target protein. Pharmacophore models confirmed their potential as selective enzyme inhibitors through 3D virtual screening.