Kiely J S, Moos W H, Pavia M R, Schwarz R D, Woodard G L
Parke-Davis Pharmaceutical Research Division, Warner-Lambert Company, Ann Arbor, Michigan 48106.
Anal Biochem. 1991 Aug 1;196(2):439-42. doi: 10.1016/0003-2697(91)90491-b.
A procedure for the qualitative assessment of inhibitory activity towards acetylcholinesterase for a given compound is described. Solutions of the compounds of interest are spotted on silica gel TLC plates in a matrix pattern. The silica gel plate is sprayed with a solution of acetylthiocholine iodide and 5,5-dithiobis(2-nitrobenzoic acid) followed by a solution of acetylcholinesterase. The enzyme reaction produces a yellow background color with inhibitor compounds exposed as white zones where color has failed to develop. The results for a test set of compounds were compared to those obtained using the standard Ellman assay procedure and found to agree for virtually all of these compounds. The conditions of silica gel plate thickness, reagent concentration, and enzyme source under which this procedure is suitable were investigated. This represents an extremely rapid method to screen large numbers of compounds to uncover new inhibitors of acetylcholinesterase and potentially other enzymes as well.
本文描述了一种用于定性评估给定化合物对乙酰胆碱酯酶抑制活性的方法。将感兴趣的化合物溶液以矩阵模式点样于硅胶TLC板上。向硅胶板喷洒碘化硫代乙酰胆碱和5,5-二硫代双(2-硝基苯甲酸)的溶液，随后再喷洒乙酰胆碱酯酶溶液。酶促反应产生黄色背景，抑制剂化合物所在区域因颜色未显而呈现为白色区域。将一组测试化合物的结果与使用标准埃尔曼测定法获得的结果进行比较，发现几乎所有这些化合物的结果都一致。研究了该方法适用的硅胶板厚度、试剂浓度和酶源条件。这是一种极其快速的方法，可用于筛选大量化合物以发现新的乙酰胆碱酯酶抑制剂，也可能发现其他酶的抑制剂。
