Development of Chemical Probes for KCNQ Potassium Channels

描述（由申请人提供）：电压门控钾通道对神经元功能至关重要。据估计，人类有160个基因编码不同但高度同源的钾通道。由于相当大的序列同源性，基因特异性通道调节剂非常罕见，但它们是研究通道功能和开发治疗方法的热门试剂。KCNQ（或Kv7）通道家族包括5个成员：KCNQ1到KCNQ5。来自患者研究和动物模型的证据表明，KCNQ表达哪怕只有25%的微小变化都可能导致癫痫等疾病。因此，化学探针将是研究电压门控钾通道结构和功能的有力分析工具，在KCNQ的情况下，这些化合物可能对治疗开发非常有价值。为了进行大规模的复合筛选，我们生成了KCNQ通道细胞系，并开发和优化了HTS-ready协议。本申请寻求使用所开发的测定法进行大规模的化合物筛选。该提案的具体目标是：1。有效地与nih指定的MLPCN中心合作，执行一个大型化合物库（bbb10万个化合物），并验证特异性激活异聚物KCNQ2/3钾通道2的先导化合物。对KCNQ1、4和5进行二次分析和反筛。3. 进行初步表征，以选择有效的先导化合物进行详细的功能分析。该项目的成功将导致鉴定新的化合物，这些化合物对开发治疗方法和研究m电流和KCNQ离子通道功能有用。

DESCRIPTION (provided by applicant): Voltage-gated potassium channels are critical for neuronal function. There are estimated 160 genes encoding different but highly homologous potassium channels in humans. Because of the considerable sequence homology, gene-specific channel modulators are very rare but they are sought-after reagents both for investigating channel function and developing therapeutics. The KCNQ (or Kv7) channel family includes five members: KCNQ1 to KCNQ5. Evidence from patient studies and animal models has shown that a small change in KCNQ expression by as little as 25% can cause disease conditions such as epilepsy. Therefore, chemical probes would be powerful analytical tools to investigate the structure and function of voltage-gated potassium channels, and in the case of KCNQ, these compounds may be very valuable for therapeutic development. To perform a large-scale compound screen, KCNQ channel cell lines have been generated and an HTS-ready protocol has been developed and optimized. The present application seeks to conduct a large scale compound screen using the developed assay. The specific aims of the proposal are: 1. To effectively work with the NIH-assigned MLPCN center to perform a large compound library (>100,000 compounds) and validate lead compounds that specifically ACTIVATE the heteromultimeric KCNQ2/3 potassium channel 2. To conduct the secondary assays and counter screen against KCNQ1, 4 and 5. 3. To perform initial characterization to allow for selection of potent lead compounds for detail functional analyses. The success of this project will lead to identification of novel compounds that are useful for developing therapeutics and investigating M-current and KCNQ ion channel function. PUBLIC HEALTH RELEVANCE: Small molecules that regulate ion flux are important tools to develop drugs to treat brain and heart diseases. They are also useful research probes to understand structure and function of ion channel proteins which mediate membrane ionic flux. This proposal is aimed at carrying out a screen of large compound library in an effort of finding novel chemical modulators for ion channels.