Multi-Angle Bioactivity Cartography for Computational Screening and Mechanistic Analysis of AChE Inhibitors From...

Acetylcholinesterase (AChE) inhibitors are crucial for the symptomatic management of Alzheimer's disease (AD), with natural products-particularly botanical sources like Yellow Gastrodia elata (YGE)-serving as promising reservoirs of such inhibitors. Nevertheless, comprehensive screening and mechanistic characterization of their inhibitory potential remain limited. This study sought to identify potent AChE inhibitors from YGE, investigate their mechanisms of action, and assess their therapeutic prospects for AD. Methodologically, an integrated approach was employed, combining ultrafiltration-liquid chromatography (UF-LC) for rapid inhibitor screening, molecular docking and dynamics simulations for mechanistic insight, two-stage high-speed countercurrent chromatography for compound isolation, enzyme kinetics to delineate inhibition modalities, and network pharmacology to uncover relevant AD-related targets. The findings identified seven active constituents with notable AChE inhibition, among which parishins A and G were obtained at high purity (98.26% and 97.26%, respectively) and exhibited mixed-type inhibition with low IC₅₀ values (0.0145 and 0.0148 mM). Molecular dynamics and network pharmacology analyses further revealed critical interactions between these compounds and key AD-related targets, including ACHE, BCHE, BACE1, and PTGS2. In summary, this work underscores the potential of YGE-sourced compounds, especially parishins A and G, as effective AChE inhibitors. The established integrative computational platform facilitates multi-dimensional bioactivity evaluation and enables hierarchical prioritization of candidate compounds, thereby offering a valuable framework for advancing natural product-derived therapeutics for AD.