Najafi_2025_ChemistrySelect_10_e00879

In this study, a new series of hexahydroacridine-1,8-dione derivatives was designed, synthesized, and evaluated as cholinesterase inhibitors for potential Alzheimer's disease (AD) therapy. The compounds were synthesized using 4-hydroxybenzaldehyde (4a4h) and vanillin (4i4p) as key precursors. Enzymatic assays revealed that vanillin-derived compounds exhibited notable butyrylcholinesterase (BChE) inhibition, with compound 4p (R1 = OMe, R2 = 4-CH3) emerging as the most potent BChE inhibitor (IC50 = 13.8 microM). Additionally, 4p demonstrated acetylcholinesterase (AChE) inhibition with an IC50 of 35.63 microM. Structureactivity relationship (SAR) analysis highlighted that the methoxy group in vanillin-derived analogs enhanced binding affinity through pp stacking and hydrogen bonding interactions. Kinetic studies identified 4p as a competitive type inhibitor of BChE (K? = 11.13 microM). Compound 4p showed approximately 65% cell viability in SH-SY5Y cells at 80 microM and did not exhibit significant toxicity at lower concentrations. Molecular docking confirmed key interactions with Trp82, His438, and Gly117, whereas molecular dynamics simulations validated the stability of the 4p-BChE complex during 100 ns. In silico analyses indicated that these compounds meet acceptable drug-likeness and pharmacokinetic criteria. Overall, the findings suggest that the hexahydroacridine-1,8-dione scaffold holds promise for developing novel anti-Alzheimer's drug candidates.