Efficient Combination of Complex Chromatography, Molecular Docking and Enzyme Kinetics for Exploration of Acetylcholinesterase Inhibitors from Poria cocos

• Published in: Molecules (Basel, Switzerland) Vol. 28; no. 3; p. 1228
• Main Authors: Wu, Tong, Hou, Wanchao, Liu, Chunming, Li, Sainan, Zhang, Yuchi
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 27-01-2023; MDPI
• Subjects: Acetylcholinesterase; acetylcholinesterase inhibitors; Affinity; Alzheimer Disease; Alzheimer's disease; Binding sites; Chinese medicine; Cholinesterase Inhibitors - analysis; Cholinesterase Inhibitors - pharmacology; Chromatography; Chromatography, High Pressure Liquid - methods; counter-current chromatography; Crystal structure; Diuretics; Drug development; Drug dosages; Efficiency; enzymatic reactions kinetics; Enzyme kinetics; Ethanol; Genetic algorithms; Herbal medicine; High performance liquid chromatography; Inflammation; Ingredients; Inhibitors; Kinetics; Ligands; Liquid chromatography; Mass spectrometry; Methods; Molecular docking; Molecular Docking Simulation; molecular docking technique; Natural products; Optimization; Poria - chemistry; Poria cocos; Proteins; Reaction kinetics; Retention; Scientific imaging; semi-preparative high−performance liquid chromatography; Ultrafiltration; Wolfiporia - chemistry; semi-preparative high−performance liquid chromatography; acetylcholinesterase inhibitors; counter-current chromatography; enzymatic reactions kinetics; molecular docking technique; Poria cocos
• ISSN: 1420-3049; 1420-3049
• DOI: 10.3390/molecules28031228

is a traditional Chinese medicinal product with the same origin as medicine and food. It has diuretic, anti-inflammatory and liver protection properties, and has been widely used in a Chinese medicine in the treatment of Alzheimer's disease (AD). This study was conducted to explore the activity screening, isolation of acetylcholinesterase inhibitors (AChEIs), and in vitro inhibiting effect of
. The aim was to develop a new extraction process optimization method based on the Matlab genetic algorithm combined with a traditional orthogonal experiment. Moreover, bio-affinity ultrafiltration combined with molecular docking was used to screen and evaluate the activity of the AChEIs, which were subsequently isolated and purified using high-speed counter-current chromatography (HSCCC) and semi-preparative high-performance liquid chromatography (semi-preparative HPLC). The change in acetylcholinesterase (AChE) activity was tested using an enzymatic reaction kinetics experiment to reflect the inhibitory effect of active compounds on AChE and explore its mechanism of action. Five potential AChEIs were screened via bio-affinity ultrafiltration. Molecular docking results showed that they had good binding affinity for the active site of AChE. Meanwhile, the five active compounds had reversible inhibitory effects on AChE: Polyporenic acid C and Tumulosic acid were non-competitive inhibitors; 3-Epidehydrotumulosic acid was a mixed inhibitor; and Pachymic acid and Dehydrotrametenolic acid were competitive inhibitors. This study provided a basis for the comprehensive utilization of
and drug development for the treatment of AD.