Subtype-Specific Small Molecule Chemical Probes for Non-neuronal KCNQ1 Potassium

用于非神经元 KCNQ1 钾的亚型特异性小分子化学探针

基本信息

批准号：
7928000
负责人：
Meng Wu
金额：
$ 4.1万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-04-23 至 2012-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7928000
关键词：
Action Potentials Arrhythmia Atrial Fibrillation Biological Assay Cardiac Cardiology Cell Line Cell physiology Cells Cellular Membrane Chemicals Chemosensitization Collaborations Complex Cyclic AMP Cystic Fibrosis Development Diarrhea Disease Ear Epithelial Epithelial Cells Etiology Family Family member Fluorescence Functional disorder Future Genus Cola Hearing Heart Heart Arrest Heart Diseases Homeostasis Hormones Intestines Investigation Ion Channel Ions Jervell-Lange Nielsen Syndrome Kidney Libraries Long QT Syndrome Lung Membrane Proteins Neurons Pharmaceutical Chemistry Pharmacologic Substance Pharmacology Play Potassium Potassium Channel Preparation Role Romano-Ward Syndrome Safety Signal Transduction Sodium Chloride Structure Sudden infant death syndrome Syndrome Testing Thallium Therapeutic Tissues Universities Validation Voltage-Gated Potassium Channel Water base hearing impairment high throughput screening improved medical schools patch clamp public health relevance salt balance scaffold small molecule tool

项目摘要

DESCRIPTION (provided by applicant): Ion channels are membrane proteins that selectively conduct ions across cellular membranes, existing in both excitable cells and non-excitable cells. Ion channels play a critical role in cellular physiology, including electrical and cellular signaling, ion homeostasis, and hormone secretion. The objective of this proposal is to find small molecule chemical probes that activate or potentiate a disease-causing potassium channel - KCNQ1. Abnormality of this channel, through co-assembly with other accessory subunits in different tissues, is thought to be causal to both cardiac diseases and epithelial cell diseases, such as long QT syndrome and cystic fibrosis. We have developed cell lines, validated the cell lines in a fluorescence-based thallium surrogate flux assay, and perform feasibility trials of high throughput screening. This proposal outlines a specific plan to conduct a >300,000-compound screen using the KCNQ1 cell line to search for specific activators/potentiators. The active compounds will then be evaluated by automated patch-clamp recording of various cell lines expressing different combinations of KCNQ1/KCNE channels, including KCNE1, KCNE2, KCNE3, KCNE4 and KCNE5 b- subunits. Because different KCNE subunits co-assemble with KCNQ1 in different tissues, specific probes developed from this proposal will therefore be useful for tissue-specific investigations. In collaboration with Dr. Gordon Tomaselli in Division of Cardiology and Dr. Mark Donowitz in Hopkins Center for Epithelial Disorders of Johns Hopkins School of Medicine, our compound validation plan includes testing of the isolated compounds in native preparations. These active compounds, with further efforts in pharmacology and medicinal chemistry, may be exploited for therapeutic remedy of cardiac arrhythmia and water and salt imbalance in epithelial tissues. PUBLIC HEALTH RELEVANCE: Non-neuronal KCNQ1 potassium channels are critical functional components of heartbeat, hearing, and water/salt balance in lung and intestinal tissues. Dysfunction of these potassium channels results in serious arrhythmia, cardiac arrest, hearing loss, diarrhea, and cystic fibrosis. There is increasing evidence for their causality for long QT syndrome, familial atrial fibrillation, Jervell and Lange-Nielsen syndrome, Romano-Ward syndrome, short QT syndrome, and sudden infant death syndrome (SIDS). Discovery of small molecule probes through this project can therefore provide tools for better understanding of these channels, improve assessment of safety concerns in future pharmaceutical development, and develop therapeutics for the above mentioned diseases.

描述（由申请人提供）：离子通道是膜蛋白，它们在可激发细胞和不可驱行的细胞中有选择性地导电跨细胞膜。离子通道在细胞生理学中起关键作用，包括电信号传导，离子稳态和激素分泌。该提案的目的是找到激活或增强引起疾病的钾通道-KCNQ1的小分子化学探针。该通道的异常通过与不同组织中的其他附件亚基共同组装，被认为是心脏病和上皮细胞疾病的因果关系，例如长QT综合征和囊性纤维化。我们已经开发了细胞系，验证了基于荧光的替代替代物通量测定法中的细胞系，并对高吞吐量筛选进行可行性试验。该提案概述了使用KCNQ1细胞系进行> 300,000个复合屏幕的特定计划，以搜索特定的激活剂/电位器。然后，将通过自动贴片钳记录各种细胞系的自动化化合物来评估，这些细胞系的各种细胞系表达了KCNQ1/KCNE通道的不同组合，包括KCNE1，KCNE2，KCNE2，KCNE3，KCNE3，KCNE4，KCNE4和KCNE5 B-亚基。由于不同的KCNE亚基与KCNQ1在不同组织中共组装，因此该提案开发的特定探针将对组织特异性研究有用。与戈登·托马塞利（Gordon Tomaselli）博士在心脏病学系和约翰·霍普金斯医学学院上皮疾病中心的Mark Donowitz博士合作，我们的复合验证计划包括对本机制剂中隔离化合物的测试。这些活性化合物在药理学和药物化学方面的进一步努力中可能被利用用于上皮组织中心律失常，水和盐失衡的治疗疗法。公共卫生相关性：非神经KCNQ1钾通道是肺和肠道组织中心跳，听力和水/盐平衡的关键功能组成部分。这些钾通道的功能障碍会导致严重的心律不齐，心脏骤停，听力丧失，腹泻和囊性纤维化。有越来越多的证据表明其因果关系综合征，家族性房颤，杰维尔和兰格·尼尔森综合征，罗曼诺 - 沃德综合征，短QT综合征和猝死综合征（SIDS）。因此，通过该项目发现小分子探针可以提供工具，以更好地理解这些渠道，改善对未来药物开发中安全问题的评估，并为上述疾病开发治疗剂。