High-Throughput screening for inhibitors of neuroinflammatory lipid production

神经炎症脂质产生抑制剂的高通量筛选

基本信息

批准号：
9104860
负责人：
BENJAMIN F CRAVATT
金额：
$ 49.33万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-02-15 至 2019-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9104860
关键词：
ABHD12 gene Animal Model Applications Grants Ataxia Auditory Behavioral Biochemical Biochemical Pathway Biological Biological Assay Brain Cataract Cell model Cells Chemicals Code Collaborations Coupled Defect Disease Drug Targeting Employee Strikes Enzymes Exhibits Future G-Protein-Coupled Receptors Generations Genes Genetic Goals Hereditary Disease Human Human Genetics Immune In Situ In Vitro Inflammatory Inherited Lead Libraries Link Lipase Lipids Lipopolysaccharides Metabolic Metabolism Methods Modeling Molecular Motor Mus Mutation Nervous system structure Neurodegenerative Disorders Neurologic Performance Pharmaceutical Chemistry Phenotype Phosphatidylserines Play Polyneuropathy Production Proteins Proteome Proteomics Research Role Serine Hydrolase Signal Transduction Solubility Source Staging Structure-Activity Relationship System Testing Therapeutic activity-based protein profiling age related base chemical property chemoproteomics disease-causing mutation hearing impairment high throughput screening human disease in vivo inhibitor/antagonist innovation interdisciplinary approach interest lymphoblast lysophosphatidylserine macrophage metabolomics motor deficit mouse model nervous system disorder neuroinflammation novel programs public health relevance screening small molecule libraries therapeutic target

项目摘要

DESCRIPTION (provided by applicant): A striking number of hereditary nervous system diseases are caused by deleterious mutations in poorly characterized enzymes from the serine hydrolase class. Over the past decade, our group has developed an innovative set of chemical proteomic and metabolomic platforms for assigning functions to uncharacterized serine hydrolases and have a special interest in applying these platforms to serine hydrolases that are causally linked by human genetics to nervous system diseases. In this grant application, we focus on the role that serine hydrolases play in the neurological disorder PHARC (polyneuropathy, hearing loss, ataxia, retinosis pigmentosa, and cataract). Specifically, we have determined that the serine hydrolase ABHD12, mutations of which cause PHARC, is a major brain lyso-phosphatidylserine (lyso-PS) lipase. ABHD12-/- mice exhibit elevated brain lyso-PS content and auditory and motor deficits coupled with heightened neuroinflammation, implicating deregulated lyso-PS signaling as a contributory factor to PHARC. Blocking the upstream enzyme(s) that produces lyso-PS could thus provide a therapeutic strategy to treat PHARC and possibly other (neuro) inflammatory diseases. We recently determined that the heretofore uncharacterized serine hydrolase ABHD16A is a major PS lipase responsible for generating lyso-PS in mammalian systems, including mouse brain and PHARC subject-derived lymphoblasts. The goal of this application is to perform a high-throughput screen of TSRI's 640,000+ compound library to identify structurally novel classes of ABHD16A inhibitors that can be progressed to selective and in vivo-active chemical probes. We have established a diverse and innovative set of biochemical, chemoproteomic, and lipidomic assays to rigorously assess the potency, selectivity, and cellular activity of ABHD16A inhibitors. We expect that the Specific Aims of this application will deliver multiple structurally distinct ABHD16A inhibitors that show suitable levels of potency (in vitro IC50 < 1 µM), selectivity (a limited number of off-targets (lss than five) across the serine hydrolase class), and cellular activity (in situ IC50 < 10 µM) for prioritization as leads for medicinal chemistry optimization into in vivo-active chemical probes. Our collaborative research team has a strong track-record of performing high-throughput screens on serine hydrolases and developing leads from these screens into selective and in vivo-active inhibitors. We are therefore well-equipped to progress the prioritized inhibitors discovered in this application toward optimized chemical probes for testing the function of ABHD16A and its lyso-PS products in PHARC and other (neuro) inflammatory disorders.

描述（由申请人提供）：大量遗传性神经系统疾病是由丝氨酸水解酶类中特征不明的酶的有害突变引起的。在过去的十年中，我们的小组开发了一套创新的化学蛋白质组学和代谢组学平台用于分配。功能对未表征的丝氨酸水解酶，并且对将这些平台应用于与人类遗传学与神经系统疾病有因果关系的丝氨酸水解酶有特别的兴趣。丝氨酸水解酶在神经系统疾病 PHARC（多发性神经病、听力损失、共济失调、视网膜色素变性和白内障）中发挥作用。 PS) 脂肪酶 ABHD12-/- 小鼠表现出脑溶血 PS 含量升高以及伴有哮喘的听觉和运动缺陷。因此，我们最近确定，阻断产生 lyso-PS 的上游酶可以提供治疗 PHARC 和可能的其他（神经）炎症性疾病的治疗策略。迄今为止未表征的丝氨酸水解酶 ABHD16A 是一种主要的 PS 脂肪酶，负责在哺乳动物系统（包括小鼠大脑和 PHARC 受试者来源）中生成溶血 PS该应用的目标是对 TSRI 的 640,000 多种化合物库进行高通量筛选，以鉴定结构新颖的 ABHD16A 抑制剂，这些抑制剂可发展为选择性和体内活性的化学探针。一套生化、化学蛋白质组学和脂质组学检测严格评估 ABHD16A 抑制剂的效力、选择性和细胞活性。该应用的目标是提供多种结构不同的 ABHD16A 抑制剂，这些抑制剂显示出适当水平的效力（体外 IC50 < 1 µM）、选择性（整个丝氨酸水解酶类中脱靶数量有限（少于 5 个））和细胞活性（原位 IC50 < 10 µM）优先作为体内活性化学探针药物化学优化的先导，我们的合作研究团队拥有良好的执行记录。对丝氨酸水解酶进行高通量筛选，并将这些筛选的先导物开发成选择性和体内活性抑制剂，因此我们有能力将本申请中发现的优先抑制剂发展为优化化学探针，以测试 ABHD16A 及其溶血的功能。 -PS 产品用于 PHARC 和其他（神经）炎症性疾病。