Real Time Screening for GIRK1/4 Channel Blockers

实时筛查 GIRK1/4 通道阻断剂

• 批准号: 8050240
• 负责人: KENNETH B WALSH
• 金额: $ 13.7万
• 依托单位: UNIVERSITY OF SOUTH CAROLINA AT COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-27 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8050240
• 关键词: Abbreviations; Acetylcholine; Acids; Amiodarone; Anti-Arrhythmia Agents; Arrhythmia; Atrial Fibrillation; Binding; Biological Assay; Carbachol; Cardiac; Cardiac Myocytes; Cell Line; Cell membrane; Cells; Dyes; Equilibrium; Experimental Designs; GTP-Binding Proteins; Gated Ion Channel; Goals; Health Care Costs; Heart; Heart Atrium; Heating; Image; Incidence; Laboratories; Libraries; Measurement; Measures; Mediating; Medical; Membrane Potentials; Methods; Modeling; Molecular; Monitor; Morbidity - disease rate; Movement; Muscarinic M2 Receptor; Muscarinics; Muscle Cells; NIH Program Announcements; Pharmaceutical Preparations; Pharmacotherapy; Phase; Potassium Channel; Preparation; Procedures; Protein Family; Proteins; Rattus; Reader; Research Project Grants; Rest; Screening procedure; Signal Transduction; South Carolina; System; Testing; Universities; Work; aging population; assay development; base; channel blockers; density; dronedarone; drug discovery; high throughput screening; inward rectifier potassium channel; ion channel blocker; member; mortality; new therapeutic target; novel; novel therapeutics; patch clamp; prevent; response; sensor; small molecule libraries; tertiapin-Q; tv watching; voltage

DESCRIPTION (provided by applicant): Atrial fibrillation (AF) is the most commonly occurring cardiac arrhythmia and is responsible for significant morbidity, mortality and health care costs. The incidence of AF is expected to increase markedly with the aging population. However, the use of conventional anti-arrhythmic drugs for treating AF has been limited due to their pro- arrhythmic actions. One novel target for AF drug therapy is the acetylcholine-activated K+ channel (IK,Ach). This channel is a member of the super-family of proteins known as the G protein-coupled inward rectifier K+ (GIRK) channels and is composed of the GIRK1/4 subunits. IK,Ach opens upon binding of acetylcholine to the cardiac muscarinic (M2) receptor and mediates vagal effects on heat rate and atrial excitation. Recent studies indicate that IK,Ach is constitutively active in AF, thus enhancing atrial excitability. The goal of this project is to develop a high throughput-screening (HTS) assay for identifying new drugs that inhibit IK,Ach. The hypothesis to be tested in this project is that membrane potential-sensitive dyes can be used as a HTS sensor in screening for IK,Ach blockers in immortalized cardiac cell lines. In Aim #1 methods for the measurement and analysis of IK,Ach, using a fluorescent imaging plate reader (FLIPR), will be established. In Aim #2 a high density HTS assay will be developed and screened with a library of ion channel blockers. New compounds identified in the assay will be tested in an isolated rat heart model of AF. Overall, the long term goal of work in the laboratory is to develop novel therapeutic strategies for treating cardiac arrhythmias. PUBLIC HEALTH RELEVANCE: Atrial fibrillation (AF) is the most commonly occurring cardiac arrhythmia and is responsible for significant morbidity, mortality and health care costs. The incidence of AF is expected to increase markedly with the aging population. The goal of this project is to identify new drugs for treating AF.

描述（由申请人提供）：房颤（AF）是最常见的心律失常，并导致显著的发病率、死亡率和医疗保健费用。随着人口老龄化，预计AF的发病率将显著增加。然而，由于其促心律失常作用，常规抗心律失常药物用于治疗AF的使用受到限制。AF药物治疗的一个新靶点是乙酰胆碱激活的K+通道（IK，Ach）。该通道是称为G蛋白偶联内向整流钾离子（GIRK）通道的蛋白质超家族的成员，由GIRK 1/4亚基组成。IK，Ach在乙酰胆碱与心脏毒蕈碱（M2）受体结合后打开，并介导迷走神经对心率和心房兴奋的影响。近年来的研究表明，心房颤动时IK、Ach具有组成性活性，从而增强心房的兴奋性。本项目的目标是开发一种高通量筛选（HTS）方法，用于鉴定抑制IK、Ach的新药。本项目的假设是，膜电位敏感染料可作为高温超导传感器用于永生化心肌细胞系中IK、Ach阻断剂的筛选。在目标#1中，将建立使用荧光成像板读取器（FLIPR）测量和分析IK、Ach的方法。在目标#2中，将开发高密度HTS测定并使用离子通道阻断剂文库进行筛选。新的化合物中确定的测定将在一个孤立的大鼠心脏模型的AF进行测试。总体而言，在实验室工作的长期目标是开发新的治疗策略，治疗心律失常。 公共卫生关系：心房纤颤（AF）是最常见的心律失常，并导致显著的发病率、死亡率和医疗保健成本。随着人口老龄化，预计AF的发病率将显著增加。该项目的目标是确定治疗AF的新药。