Giannetti Anthony M, Koch Bruce D, Browner Michelle F
Roche Palo Alto LLC, Palo Alto, CA 94304, USA.
J Med Chem. 2008 Feb 14;51(3):574-80. doi: 10.1021/jm700952v. Epub 2008 Jan 9.
Promiscuous binders achieve enzyme inhibition using a nonspecific aggregation-type binding mechanism to proteins. These compounds are a source of false-positive hits in biochemical inhibition assays and should be removed from screening hit lists because they are not good candidates to initiate medicinal chemistry programs. We introduce a robust approach to identify these molecules early in the lead generation process using real time surface plasmon resonance based biosensors to observe the behavior of the binding interactions between promiscuous compounds and proteins. Furthermore, the time resolution of the assay reveals a number of distinct mechanisms that promiscuous compounds employ to inhibit enzyme function and indicate that the type of mechanism can vary depending on the protein target. A classification scheme for these compounds is presented that can be used to rapidly characterize the hits from high-throughput screens and eliminate compounds with a nonspecific mechanism of inhibition.
混杂结合剂通过一种与蛋白质的非特异性聚集型结合机制实现酶抑制。这些化合物是生化抑制试验中假阳性命中的来源，应从筛选命中列表中去除，因为它们不是启动药物化学项目的理想候选物。我们引入了一种强大的方法，在先导化合物生成过程的早期使用基于表面等离子体共振的实时生物传感器来识别这些分子，以观察混杂化合物与蛋白质之间结合相互作用的行为。此外，该检测的时间分辨率揭示了混杂化合物用于抑制酶功能的许多不同机制，并表明机制类型可能因蛋白质靶点而异。本文提出了一种针对这些化合物的分类方案，可用于快速表征高通量筛选中的命中物，并消除具有非特异性抑制机制的化合物。
