目的 采用亲和超滤-液质联用技术（affinity ultra-filtration-liquid chromatography-mass spectrometry，AUF-LC-MS）靶向筛选特色药材多裂骆驼蓬中的乙酰胆碱酯酶（acetylcholinesterase，AChE）抑制剂，并探究其抑制机制。方法 采用Ellman法评价多裂骆驼蓬不同极性部位提取物的AChE抑制活性，采用AUF-LC-MS对活性成分精准筛选和鉴定。利用酶抑制动力学、分子对接和分子动力学模拟探究活性成分对AChE抑制作用机制。结果 从二氯甲烷和正丁醇2个部位共筛选出7个活性成分，包括1个黄酮类和6个生物碱。酶抑制动力学显示7种成分分属于可逆竞争型（蒙花苷）、可逆非竞争型（鸭嘴花碱、鸭嘴花酮碱、哈尔酚）和可逆混合型（脱氧鸭嘴花碱、脱氧鸭嘴花酮碱、去氢骆驼蓬碱）3种抑制类型。分子对接显示，活性成分与AChE有良好的结合性能，不同抑制剂的结合位点和作用力存在差异。分子动力学模拟表明体系已达稳定，在动态结合过程中，抑制剂与AChE存在与分子对接过程中类似的强且稳定的相互作用。结论 相关研究为诠释多裂骆驼蓬防治阿尔茨海默病提供科学依据，为发现高效低毒AChE抑制剂提供新参考。

Objective To screen acetylcholinesterase (AChE) inhibitors from the specific medicinal herbs Peganum multisectum by affinity ultra-filtration-liquid chromatography-mass spectrometry (AUF-LC-MS), and to explore their inhibition mechanisms. Methods The AChE inhibitory activities of various polar fractions of P. multisectum were evaluated by Ellman’s test, and the active components were precisely screened by AUF-LC-MS. The action mechanisms of the active components on AChE were investigated by enzyme inhibition kinetics, molecular docking and molecular dynamics simulations. Results A total of seven active components, including one flavonoid and six alkaloids, were identified from the dichloromethane and n-butanol fractions. Enzyme inhibition kinetics suggested that these components belonged to three inhibition types: reversible competitive inhibition (linarin), non-competitive (vasicine, vasicinone and harmol), and reversible mixed inhibition (deoxyvasicine, deoxyvasicinone and harmaline). Molecular docking results revealed that all compounds exhibited good affinities to AChE. And the binding sites and interaction forces of these inhibitors with AChE differed. Molecular dynamics simulation indicated that the system had reached a stable state, and the inhibitors had strong and stable interactions with AChE. Conclusions This study provided a scientific basis for elucidating the potential of P. multisectum in the prevention and treatment of Alzheimer’s disease and offered a new reference for the discovery of high-efficiency AChE inhibitors.

R284.1

国家自然科学基金项目（82374161）；中央高校基本科研业务费-中央民族大学交叉学科研究专项（2024JCYJ09）；北京市级大学生科学研究与创业行动计划项目（BEIJ2024110151）