Feng Brian Y, Simeonov Anton, Jadhav Ajit, Babaoglu Kerim, Inglese James, Shoichet Brian K, Austin Christopher P
Department of Pharmaceutical Chemistry & Graduate Group in Chemistry and Chemical Biology, 1700 4th Street, University of California San Francisco, San Francisco, California 94158-2330, USA.
J Med Chem. 2007 May 17;50(10):2385-90. doi: 10.1021/jm061317y. Epub 2007 Apr 21.
High-throughput screening (HTS) is the primary technique for new lead identification in drug discovery and chemical biology. Unfortunately, it is susceptible to false-positive hits. One common mechanism for such false-positives is the congregation of organic molecules into colloidal aggregates, which nonspecifically inhibit enzymes. To both evaluate the feasibility of large-scale identification of aggregate-based inhibition and quantify its prevalence among screening hits, we tested 70,563 molecules from the National Institutes of Health Chemical Genomics Center (NCGC) library for detergent-sensitive inhibition. Each molecule was screened in at least seven concentrations, such that dose-response curves were obtained for all molecules in the library. There were 1274 inhibitors identified in total, of which 1204 were unambiguously detergent-sensitive. We identified these as aggregate-based inhibitors. Thirty-one library molecules were independently purchased and retested in secondary low-throughput experiments; 29 of these were confirmed as either aggregators or nonaggregators, as appropriate. Finally, with the dose-response information collected for every compound, we could examine the correlation between aggregate-based inhibition and steep dose-response curves. Three key results emerge from this study: first, detergent-dependent identification of aggregate-based inhibition is feasible on the large scale. Second, 95% of the actives obtained in this screen are aggregate-based inhibitors. Third, aggregate-based inhibition is correlated with steep dose-response curves, although not absolutely. The results of this screen are being released publicly via the PubChem database.
高通量筛选（HTS）是药物发现和化学生物学中鉴定新先导化合物的主要技术。不幸的是，它容易出现假阳性结果。产生此类假阳性的一种常见机制是有机分子聚集成胶体聚集体，这些聚集体会非特异性地抑制酶。为了评估大规模鉴定基于聚集体的抑制作用的可行性并量化其在筛选结果中的发生率，我们测试了来自美国国立卫生研究院化学基因组学中心（NCGC）文库的70563个分子的去污剂敏感性抑制作用。每个分子都在至少七种浓度下进行筛选，从而获得文库中所有分子的剂量反应曲线。总共鉴定出1274种抑制剂，其中1204种明确对去污剂敏感。我们将这些鉴定为基于聚集体的抑制剂。从文库中独立购买了31个分子，并在二次低通量实验中重新进行测试；其中29个被相应地确认为聚集体或非聚集体。最后，利用为每种化合物收集的剂量反应信息，我们可以研究基于聚集体的抑制作用与陡峭剂量反应曲线之间的相关性。这项研究得出了三个关键结果：第一，基于去污剂的基于聚集体的抑制作用的大规模鉴定是可行的。第二，在此筛选中获得的活性物质中有95％是基于聚集体的抑制剂。第三，基于聚集体的抑制作用与陡峭的剂量反应曲线相关，尽管不是绝对相关。该筛选结果正在通过PubChem数据库公开发布。
