Small Molecule Inhibitors of Phospholipase C from a Novel High-throughput Screen
Small Molecule Inhibitors of Phospholipase C from a Novel High-throughput Screen
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DOI：
10.1074/jbc.m112.422501
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发表时间：
2013-02-22
影响因子：
4.8
通讯作者：
Zhang, Qisheng
中科院分区：
文献类型：
--
作者：
Huang, Weigang;Barrett, Matthew;Zhang, Qisheng
磷脂酶C(PLC)同工酶是重要的信号分子，但很少有小分子调节剂可用于药理调节其功能。为了建立一种鉴定新型PLC抑制剂的通用方法，我们建立了一种基于荧光底物报告基因WH-15的高通量检测方法。该方法灵敏度高，重复性好：对6280种化合物的化学文库进行筛选，鉴定出三种新的PLC抑制剂，它们在体外用纯化的PLC同工酶在两种不同的检测形式下显示出强大的活性。三种抑制剂中的两种也抑制了G蛋白偶联受体刺激的完整细胞系统中PLC的活性。这些结果证明了高通量分析在筛选大量小分子以识别新的PLC调节子方面的能力。有效和选择性的PLC调节剂最终将有助于剖析PLC在细胞过程中的作用，并为开发治疗磷脂酶活性异常引起的疾病的药物提供先导化合物。
Phospholipase C (PLC) isozymes are important signaling molecules, but few small molecule modulators are available to pharmacologically regulate their function. With the goal of developing a general approach for identification of novel PLC inhibitors, we developed a high-throughput assay based on the fluorogenic substrate reporter WH-15. The assay is highly sensitive and reproducible: screening a chemical library of 6280 compounds identified three novel PLC inhibitors that exhibited potent activities in two separate assay formats with purified PLC isozymes in vitro. Two of the three inhibitors also inhibited G protein-coupled receptor-stimulated PLC activity in intact cell systems. These results demonstrate the power of the high-throughput assay for screening large collections of small molecules to identify novel PLC modulators. Potent and selective modulators of PLCs will ultimately be useful for dissecting the roles of PLCs in cellular processes, as well as provide lead compounds for the development of drugs to treat diseases arising from aberrant phospholipase activity.