Di Giovanni Saviana, Borloz Aline, Urbain Aurélie, Marston Andrew, Hostettmann Kurt, Carrupt Pierre-Alain, Reist Marianne
LCT-Pharmacochimie, Ecole de Pharmacie Genève-Lausanne, Section des Sciences Pharmaceutiques, Quai Ernest-Ansermet 30, CH-1211 Genève 4, Switzerland.
Eur J Pharm Sci. 2008 Feb 5;33(2):109-19. doi: 10.1016/j.ejps.2007.10.004. Epub 2007 Oct 30.
Acetylcholinesterase inhibitors (AChEI) are currently still the best available pharmacotherapy for Alzheimer patients. Successful screening for new AChEI relies on effective and fast assays. Two colorimetric screening assays frequently used to search for new AChEI, namely a thin layer chromatography (TLC) assay with Fast Blue B salt as reagent and a 96-well plate assay based on Ellman's method, were compared. For the majority (83%) of the 138 test compounds of natural and synthetic origin, the results obtained with the two assays converged and both screening assays were considered suitable for the generation of new hits. Fifteen percent of investigated compounds were classified as active with the microplate assay but were shown to be inactive by TLC and about 2% were measured active by TLC but showed to be inactive with the microplate assay. These divergences were not due to the main differences between the experimental protocols of the two screening assays, namely the different colorimetric methods and pre-incubation of test compounds with acetylcholinesterase (AChE). They might be explained by the interaction of either AChE or test compounds with the silica of the TLC plates, resulting in an altered affinity of the enzyme for the compounds.
乙酰胆碱酯酶抑制剂（AChEI）目前仍是治疗阿尔茨海默病患者的最佳可用药物疗法。成功筛选新型AChEI依赖于有效且快速的检测方法。对常用于寻找新型AChEI的两种比色筛选方法进行了比较，即使用固蓝B盐作为试剂的薄层色谱（TLC）法和基于埃尔曼方法的96孔板法。对于138种天然和合成来源的测试化合物中的大多数（83%），两种检测方法得到的结果一致，且两种筛选方法均被认为适用于产生新的活性物质。15%的研究化合物在微孔板检测中被归类为有活性，但经TLC检测显示无活性，约2%的化合物经TLC检测有活性，但在微孔板检测中显示无活性。这些差异并非由于两种筛选方法实验方案的主要差异，即不同的比色方法以及测试化合物与乙酰胆碱酯酶（AChE）的预孵育。它们可能是由AChE或测试化合物与TLC板硅胶的相互作用所解释，从而导致酶对化合物的亲和力发生改变。
