Liu_2025_Bioorg.Chem_161_108551

Cholinesterase inhibitors (ChEIs) are widely utilized for the symptomatic management of Alzheimer's disease (AD) by enhancing acetylcholine levels to improve cognitive function. Concurrently, neuroinflammation has emerged as a critical factor in AD progression, necessitating therapies that address this pathology. In this study, we designed and synthesized a novel bifunctional cholinesterase inhibitor, (E)-4-(2-(3-(benzyloxy)-4-oxo-4H-pyran-2-yl) vinyl)-1,2-phenylene bis(ethyl(methyl)carbamate) (D40), which combines potent cholinesterase inhibition with robust anti-neuroinflammatory activity. D40 demonstrated potent inhibition of human butyrylcholinesterase (hBuChE), with an IC50 value of 0.59 +/- 0.03 microM, significantly outperforming Rivastigmine (IC50 = 3.70 +/- 0.96 microM). Molecular docking and molecular dynamics simulations confirmed a stable and selective binding of D40 to the BuChE active site, underpinning its inhibitory profile. Additionally, D40 exhibited strong anti-inflammatory effects, with an IC50 value of 4.55 +/- 0.78 microM for suppressing nitric oxide production and demonstrated excellent blood-brain barrier permeability. In vivo studies in aged 5 x FAD mice revealed that D40 significantly reduced neuroinflammation by suppressing pro-inflammatory cytokines and glial activation. Furthermore, D40 mitigated Abeta deposition, promoted neuronal survival, and improved cognitive deficits, while demonstrating a favorable safety profile in acute toxicity evaluations. These findings highlight D40 as a dual-function ChEI capable of providing symptomatic relief and modulating neuroinflammatory pathways associated with AD. With its enhanced cholinesterase inhibition and anti-inflammatory properties, D40 emerges as a promising candidate for the treatment of advanced stages of AD. Acetylcholine deficiency and neuroinflammation as drivers of Alzheimer's disease dually intervened by Compound D40.