Affinity ultrafiltration screening, multi-target extraction, and comprehensive activity verification of acetylcholinesterase inhibitors in the food additive Corydalis yanhusuo

Abstract Corydalis yanhusuo possesses strong medicinal value and is used in certain regions for soup making, wine frying, vinegar processing, and other edible processes. The anti-Alzheimer’s disease (AD) traditional Chinese patent medicine "Yangxue Qingnao Granules" contains C. yanhusuo. To explore the anti-dementia components of C. yanhusuo, this study used acetylcholinesterase (AChE) as the pointcut and screened, extracted, separated, and evaluated the biological activity of AChE inhibitors in C. yanhusuo. For the first time, affinity ultrafiltration combined with a matrix comprehensive evaluation of AChE inhibitor activity was used, and activity prediction analysis was conducted using a binding-degree growth trend fitting diagram. The response surface combined with pairwise Euclidean distances and Non-dominated Sorting Genetic Algorithm III technology was employed to classify various compounds, thereby optimizing the extraction of multiple active ingredients. Semi-preparative high-performance liquid chromatography was used to separate and purify the active compounds. The inhibitory effects and mechanisms of action of the active ingredients on AChE were verified through enzymatic reaction kinetics, and the potential anti-AD effects of the active ingredients were verified through network pharmacology. Finally, the differences in compound activity were comprehensively evaluated for the first time using a Pearson correlation matrix. The results revealed that six compounds in C. yanhusuo exhibited significant binding affinities for the AChE active site. This study provides valuable insights into the active compounds of C. yanhusuo and their potential anti-AD properties, promotes the development of AChE inhibitors from food and medicines, and provides an approach to reduce the side effects associated with synthetic drugs. This study provides a valuable reference for the exploration and development of functional materials from various medicinal resources.