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

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

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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（由适用提供）：遗传神经系统疾病数量引人注目，是由于丝氨酸水解酶类别的酶中删除的突变引起的。在过去的十年中，我们的小组开发了一组创新的化学蛋白质组学和代谢组平台，用于将功能分配给未表征的丝氨酸水解酶，并特别感兴趣地将这些平台应用于人类遗传学与神经系统疾病有关的丝氨酸水解酶。在此赠款应用中，我们重点介绍丝氨酸水解酶在神经系统疾病PHARC（多神经病，听力损失，共济失调，视网膜病，色素色素和白内障）中起的作用。具体而言，我们已经确定丝氨酸水解酶ABHD12（引起PHARC的突变）是主要的脑溶菌磷脂酰丝氨酸（Lyso-PS）脂肪酶。 ABHD12 - / - 小鼠暴露了升高的脑溶解液-PS含量和听觉和电动机，并定义了神经炎症的增强，暗示了对PHARC的促成因素的放松调节的Lyso-PS信号传导。因此，阻止产生溶血-PS的上游酶可以提供治疗PHARC和其他可能（神经）炎症性疾病的治疗策略。我们最近确定迄今未表征的丝氨酸水解酶ABHD16A是负责在哺乳动物系统中产生溶血性PS的主要PS脂肪酶，包括小鼠脑和PHARC受试者衍生的淋巴细胞。该应用程序的目的是执行TSRI 640,000+复合库的高通量屏幕，以识别可以发展为选择性和体内活性化学问题的结构新颖的ABHD16A抑制剂。我们已经建立了一套多样性和创新的生化，化学蛋白质组学和脂肪组学评估，以严格评估ABHD16A抑制剂的效力，选择性和细胞活性。我们预计，本应用的具体目的将提供多种在结构上不同的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（少量的脱靶（LS小于5）（LSS小于5），整个丝氨基水解酶类别）和细胞活性（以便在situ IC50 <10 µm）中进行了化学化的化学作用，以实现优先级别的化学作用。我们的协作研究团队拥有在丝氨酸水解酶上执行高通量屏幕，并将这些屏幕从这些屏幕开发为选择性和体内活性抑制剂的轨道。因此，我们有良好的能力来进步本应用中发现的优先抑制剂，以优化化学问题，以测试ABHD16A及其在PHARC和其他（Neuro）炎症性疾病中的Lyso-PS产品的功能。