High throughput screening for small molecule inhibitors of heparin-induced tau fibril formation
High throughput screening for small molecule inhibitors of heparin-induced tau fibril formation
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DOI：
10.1016/j.bbrc.2007.03.056
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发表时间：
2007-06-22
影响因子：
3.1
通讯作者：
Lee, Virginia M. -Y.
中科院分区：
文献类型：
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作者：
Crowe, Alex;Ballatore, Carlo;Lee, Virginia M. -Y.
通过使用肝素诱导的tau纤化试验的高通量筛选(HTS)来查询包含51,000个化合物的文库。用细菌表达的重组tau片段K18进行HTS，并用硫代黄素T荧光法监测反应。符合HTS中选择标准的HITS在一组分析中进一步评估，该小组旨在(A)确认初始结果和(B)识别非特定机制或依赖于分析的伪影可能产生的假阳性。两个2，3-二(呋喃-2-基)喹恶啉被确认为牛磺酸纤化的抑制剂，IC(50)S在低微摩尔范围(1-3µM)。在假阳性中，嘧啶并三嗪类、苯并呋喃类、卟啉类和蒽醌类化合物通过催化氧化还原循环产生过氧化氢来抑制tau的纤化，这是由于分析中的还原剂二硫苏糖醇(DTT)所致。这项研究描述了tau纤化抑制剂HTS的重点策略，这些药物与阿尔茨海默病和相关tauopathy的药物发现相关。(C)2007 Elsevier Inc.保留所有权利。
A library of 51,000 compounds was interrogated by high throughput screening (HTS) using a heparin-induced tau fibrillization assay. HTS was conducted with bacterially expressed recombinant tau fragment K18 and the reaction was monitored by thioflavine T fluorescence. Hits meeting criteria set for selection in HTS were further evaluated in a panel of assays designed (a) to confirm the initial results and (b) to identify possible false positives arising from non-specific mechanisms or assay-dependent artifacts. Two 2,3-di(furan-2-yl)-quinoxalines were confirmed as inhibitors of tau fibrillization with IC(50)s in the low micromolar range (1-3 mu M). Among false positive hits, members of the pyrimidotriazines, benzofurans, porphyrins, and anthraquinone, inhibited tau fibrillization by generating peroxides via catalytic redox cycles due to the reducing agent dithiothreitol (DTT) in the assay. This study delineates focused strategies for HTS of tau fibrillization inhibitors that are relevant to drug discovery for Alzheimer's disease and related tauopathies. (c) 2007 Elsevier Inc. All rights reserved.