Development of a high-throughput screen for inhibitors of water permeability thro

开发水渗透性抑制剂的高通量筛选

• 批准号: 7325796
• 负责人: Marc F Pelletier
• 金额: $ 30万
• 依托单位: AEROMICS, INC
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-01-15 至 2009-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7325796
• 关键词: Adverse effects; Age; Altitude Sickness; American; American Heart Association; Bacterial Meningitis; Blood - brain barrier anatomy; Blood Vessels; Brain; Brain Edema; Brain Injuries; Carbon Dioxide; Cause of Death; Cells; Cerebral Edema; Cerebrum; Cessation of life; Clinical; Condition; Development; Drug Delivery Systems; Edema; Electrophysiology (science); Experimental Models; Gases; Goals; Human; Inflammation; Intracerebral Pressures; Ischemia; Ischemic Stroke; Knockout Mice; Left; Libraries; Medical; Optical Methods; Optics; Outcome; Oxygen; Pathology; Pathway interactions; Patients; Perfusion; Permeability; Pharmaceutical Preparations; Phase; Phase II Clinical Trials; Proteins; Recurrence; Research; Small Business Funding Mechanisms; Small Business Innovation Research Grant; Spinal Cord; Spinal cord injury; Stroke; Structure; Swelling; Technology; Testing; Tissues; Traumatic Brain Injury; Up-Regulation; Water; Work; aquaporin 4; cost; day; disability; high throughput screening; improved; inhibitor/antagonist; injured; retinal ischemia; scale up; statistics; water channel

DESCRIPTION (provided by applicant): Cerebral edema (and its counterpart in the spinal cord) is a major contributor to the damage caused by stroke, traumatic brain injury, traumatic spinal-cord injury, and bacterial meningitis. Stroke alone is the third leading cause of death in the USA and the leading cause of disability. Unfortunately, only a few treatment options are available for cerebral and spinal-cord edema, and these are of limited efficacy. Central in the development of cerebral and spinal-cord edema is the H2O channel aquaporin-4 (AQP4) in the astrocytic endfeet that envelope CNS blood vessels at the blood-brain barrier (BBB). AQP4 is upregulated by ischemia, traumatic brain injury, and a diverse array of other CNS pathologies associated with inflammation. But, paradoxically, AQP4-null mice do substantially better than their wild-type counterparts in experimental models of ischemic stroke, retinal ischemia, and bacterial meningitis, making AQP4 an attractive drug target. A unique aspect of the present proposal is the recognition that the AQPs are bifunctional proteins that are not only permeable to water, but also to the dissolved gases O2 and CO2. Indeed, the upregulation of AQP4 in the aforementioned pathologies may have the beneficial effect of maximizing O2 permeability across the BBB, with the side effect of increasing osmotic water permeability and predisposing to cerebral edema. The long-term goal of Aeromics is to develop a drug that treats cerebral edema by blocking the water permeability of AQP4, leaving the gas permeability intact. The goals of this SBIR application are to: (1) Develop high-throughput screens (HTSs) for inhibitors of the water permeability of AQP4, using optical methods to assess the consequences of osmotically induced cell swelling. (2) Establish counterscreens for water and gas permeabilities, using biophysical approaches founded on electrophysiology and optics. These counterscreens will identify: (a) false positives from the HTS, (b) compounds that block gas as well as H2O permeability, and (c) compounds that reduce the water permeability of AQPs other than AQP4. (3) Perform pilot high-throughput screens using the MicroSource GenPlus 960 and the Maybridge 20k libraries, and identify some core structures. The proposed work will prepare Aeromics for a Phase II study, in which the HTS would be greatly expanded to identify several core structures that would be developed into leads for new drugs to treat cerebral edema. Swelling of the brain (cerebral edema) and spinal cord contributes in a major way to the damage caused by stroke, traumatic brain injury, traumatic spinal-cord injury, and bacterial meningitis. Unfortunately, only a few treatment options are available for cerebral and spinal-cord edema, and these are of limited efficacy. The goal of this project is to develop the technology to identify new drugs for treating cerebral edema while maintaining the delivery of oxygen to the brain.

描述（由申请人提供）：脑水肿（及其在脊髓中的对应物）是中风、创伤性脑损伤、创伤性脊髓损伤和细菌性脑膜炎引起的损伤的主要原因。中风是美国第三大死亡原因，也是导致残疾的主要原因。不幸的是，只有少数治疗方案可用于脑和脊髓水肿，这些都是有限的疗效。脑和脊髓水肿发展的中心是在血脑屏障（BBB）处包裹CNS血管的星形胶质细胞终足中的H2O通道水通道蛋白-4（AQP 4）。AQP 4被局部缺血、创伤性脑损伤和多种与炎症相关的其他CNS病理上调。但是，矛盾的是，AQP 4-null小鼠在缺血性中风，视网膜缺血和细菌性脑膜炎的实验模型中比野生型小鼠表现得更好，使AQP 4成为一个有吸引力的药物靶点。本建议的一个独特的方面是认识到水通道蛋白是双功能蛋白质，不仅对水是可渗透的，而且溶解的气体O2和CO2。事实上，在上述病理中AQP 4的上调可能具有使O2穿过BBB的渗透性最大化的有益效果，具有增加渗透水渗透性和诱发脑水肿的副作用。Aeromics的长期目标是开发一种药物，通过阻断AQP 4的水渗透性来治疗脑水肿，使气体渗透性保持完整。该SBIR应用的目标是：（1）开发用于AQP 4的水渗透性抑制剂的高通量筛选（HTS），使用光学方法来评估毒性诱导的细胞溶胀的后果。(2)建立水和气体渗透性的counterscreens，使用生物物理学方法建立在电生理学和光学。这些反筛选将识别：（a）HTS的假阳性，（B）阻断气体和H2O渗透性的化合物，以及（c）降低除AQP 4以外的AQP的水渗透性的化合物。(3)使用MicroSource GenPlus 960和Maybridge 20 k文库进行中试高通量筛选，并鉴定一些核心结构。拟议的工作将为第二阶段研究准备Aeromics，其中HTS将被大大扩展，以确定几个核心结构，这些结构将被开发成治疗脑水肿的新药。脑肿胀（脑水肿）和脊髓肿胀是导致中风、创伤性脑损伤、创伤性脊髓损伤和细菌性脑膜炎的主要原因。不幸的是，只有少数治疗方案可用于脑和脊髓水肿，这些都是有限的疗效。该项目的目标是开发技术，以确定治疗脑水肿的新药，同时保持向大脑输送氧气。