Dual-acting flavonol analogues as cholinesterase inhibitors and anticonvulsant agents: integrated in vitro, in vivo, and in silico studies.

Neurological disorders such as Alzheimer's disease and epilepsy are complex, multifactorial conditions that require therapeutic agents capable of acting through multiple mechanisms. In the present study, a series of structurally diverse flavonol analogues was synthesized and evaluated for their dual cholinesterase inhibitory and anticonvulsant potential through integrated in vitro, in vivo, and in silico approaches. The compounds were screened for acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitory activities using standard spectrophotometric assays. Selected derivatives demonstrating significant enzyme inhibition were further assessed for anticonvulsant efficacy in maximal electroshock seizure (MES) and pentylenetetrazole (PTZ)-induced seizure models. Several compounds exhibited noteworthy inhibition of both cholinesterase enzymes, with the most active derivatives displaying low micromolar potency. In anticonvulsant studies, the leading compounds produced dose-dependent protection against electrically and chemically induced seizures, significantly reducing seizure severity, prolonging seizure latency, and increasing protection rates in both experimental models. To the best of our knowledge, this is among the first reports evaluating the anticonvulsant activity of this specific flavonol scaffold in both MES and PTZ models. Computational investigations, including induced-fit docking, MM-GBSA binding free-energy calculations, molecular dynamics simulations, ADME prediction, and density functional theory (DFT) analyses, revealed favorable enzyme-binding interactions, stable ligand-protein complexes, and acceptable drug-like characteristics. The convergence of biological and computational findings identified compound 8 as the most promising multifunctional candidate, exhibiting potent dual cholinesterase inhibition, pronounced anticonvulsant activity, and sustained target engagement. These results demonstrate that flavonol-based scaffolds represent attractive lead structures for the development of multifunctional therapeutic agents targeting neurodegenerative disorders and epilepsy.