微生物来源的乙酰胆碱酯酶抑制剂N98-1021A的研究

N98-1021 A,a selective acetylcholinesterase inhibitors derived from microorganisms

本研究分别以丁酰硫代胆碱和二硫二硝基苯甲酸为底物和显色剂 ,建立了一种准确、快速的高通量筛选微生物代谢产物来源的抑制剂的体外筛选模型。利用此方法从 2 14 1株放线菌中筛选到一株阳性菌株N98 10 2 1,该菌株的发酵液经有机溶剂提取、ODS柱层析及HPLC制备分离 ,得到一个活性化合物N98 10 2 1A ,该化合物对乙酰胆碱酯酶的IC50 为 2 0 μmol/L ,经酶抑制动力学分析 ,该化合物为乙酰胆碱酯酶的非竞争性可逆抑制剂。通过对该化合物的紫外、红外、质谱、核磁等理化分析 ,得知该化合物与terferol结构相同。但该化合物对乙酰胆碱脂酶的抑制活性至今未见报道。

A rapid screening method using butylthiocholine as the substrate and dithiobisnitrobenzoate as the chromogenic agent was established for screening AChE inhibitors derived from microorganisms. In the course of screening from 2141 strains of microorganisms, one culture broth N98-1021 was selected as candidates with active principle for further experiments. The broth was subjected to solvent extraction,ODS column chromatographic and preparative ODS HPLC to get an active compound named N98-1021 A, which showed strong inhibitory activity against human AChE with IC 50 of 0.02mmol/L. In vitro studies on the inhibitory modes of N98-1021 A, indicated that the inhibition was noncompetitive reversible manner. The UV, IR, MS and NMR data showed that N98-1021 A was identical with terferol.