Assay Development and Discovery of LRRK2 Inhibitors for Parkinson Disease

帕金森病 LRRK2 抑制剂的检测开发和发现

基本信息

批准号：
7993886
负责人：
SHU G. CHEN
金额：
$ 23.55万
依托单位：
CASE WESTERN RESERVE 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：
2010-07-01 至 2012-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7993886
关键词：
Adopted Affect Allosteric Site Alzheimer&apos s Disease Animal Model Animals Baculovirus Expression System Behavior Behavioral Biological Assay Biology Brain Diseases Caenorhabditis elegans Chemicals Data Detection Development Dimerization Disease Disease Progression Diversity Library Docking Dopamine Drug Delivery Systems Drug Industry Drug effect disorder Fluorescence GTP Binding Genetic Guanosine Triphosphate Phosphohydrolases High Prevalence Impairment In Vitro Individual Invertebrates Investigation Lead Length Libraries Link Locomotion Measures Mediating Modeling Modification Molecular Target Monitor Morphology Movement Disorders Mutation Nematoda Nerve Degeneration Neurodegenerative Disorders Neurons Parkinson Disease Pathogenesis Patients Pharmaceutical Preparations Pharmacologic Substance Phenotype Phosphorylation Phosphotransferases Play Population Preclinical Drug Evaluation Prevention Principal Investigator Property Protein Kinase Proteins Recombinants Research Role Screening procedure Site Staging Structure System Testing Therapeutic Transgenic Organisms Translating Universities Validation age related aged assay development base combat cytotoxicity design dopaminergic neuron drug candidate drug development drug discovery drug efficacy drug market high throughput screening in vitro Assay in vitro activity in vivo inhibitor/antagonist kinase inhibitor leucine-rich repeat kinase 2 mutant novel novel therapeutics prevent public health relevance small molecule small molecule libraries sound therapeutic development therapeutic target

项目摘要

DESCRIPTION (provided by applicant): The overall objective of our application is to translate latest research findings about LRRK2, a key player in PD pathogenesis, into the drug discovery pipeline for novel PD therapeutics. Mutations in leucine-rich repeat kinase 2 (LRRK2) are the most frequent cause thus far identified in both familial and sporadic PD. Since mutant LRRK2 proteins cause hyper-activation of LRRK2 in vitro and cytotoxicity in neurons, therapeutics targeting LRRK2 is likely to be highly effective for PD. We also reason that LRRK2 represents a rare opportunity for therapeutic development since it contains two druggable enzymatic modules (kinase domain and GTPase domain), potentially providing multiple sites for drug action. We propose to employ two complementary approaches to PD drug development targeting LRRK2. Specific Aim 1 is to use recombinant full-length LRRK2 for high throughput screening of existing drugs and additional chemical libraries for kinase inhibitors and for GTP binding blockers that will prevent LRRK2 activation through distinct mechanisms of action. Specific Aim 2 is to utilize a newly developed C. elegans model of PD for in vivo assays of LRRK2 inhibitors. Recent studies have demonstrated that C. elegans can serve as a powerful whole animal model for early-stage drug screening and validation. We have recently established transgenic C. elegans animals expressing mutant LRRK2 that manifest easily quantifiable phenotype of neurodegeneration and behavior impairment relevant to PD. This C. elegans model of LRRK2 linked PD will be used to validate and characterize the promising LRRK2 inhibitory compounds from the in vitro chemical screens. The resulting candidates of LRRK2 inhibitors may be further optimized for better efficacy and drug-like activities. A multi-disciplinary team consisting of two principal investigators and several collaborators with expertise in LRRK2 pathobiology, C. elegans biology, and drug discovery has been established. As a result, preliminary data have been generated to provide evidence that our proposed investigation is based on sound scientific principles. We believe that our proposed studies will likely facilitate the discovery of potential drug candidates targeting LRRK2, and upon further validation in mammalian models may lead to novel therapeutics for the treatment and prevention of PD. PUBLIC HEALTH RELEVANCE: Parkinson disease is a devastating and fatal brain disorder affecting many people in US. The current therapeutic options for PD are very limited and no cure exists for the disease. Because LRRK2 mutations are thus far the most frequent cause for both familial and sporadic PD, the LRRK2 based drugs will likely have a very broad impact in benefiting all PD patients. The significance of our proposed studies will be the possibility of discovering novel drug leads for the treatment and prevention of PD.

描述（由申请人提供）：我们申请的总体目的是将有关PD发病机理的关键参与者LRRK2的最新研究结果转化为新型PD Therapeutics的药物发现管道。富含亮氨酸的重复激酶2（LRRK2）中的突变是迄今为止在家族性和零星PD中鉴定出的最常见原因。由于突变的LRRK2蛋白在体外引起LRRK2的过度激活和神经元中的细胞毒性，因此靶向LRRK2的疗法可能对PD非常有效。我们还认为，LRRK2代表了治疗发育的难得机会，因为它包含两个可药物鉴定酶模块（激酶结构域和GTPase结构域），可能会提供多个用于药物作用的部位。我们建议采用两种互补方法来针对LRRK2的PD药物开发。具体目的1是使用重组全长LRRK2进行现有药物的高吞吐量筛选，并为激酶抑制剂和GTP结合阻滞剂提供其他化学文库，从而通过不同的作用机制来防止LRRK2激活。具体目的2是利用新开发的PD秀丽隐杆线虫模型进行LRRK2抑制剂的体内测定。最近的研究表明，秀丽隐杆线虫可以作为早期药物筛查和验证的强大全动物模型。我们最近建立了表达突变体LRK2的转基因秀丽隐杆线虫动物，这些动物易于量化与PD相关的神经变性和行为障碍的表型。 LRRK2链接的PD的秀丽隐杆线虫模型将用于验证和表征来自体外化学筛查的有希望的LRRK2抑制化合物。 LRRK2抑制剂的最终候选者可能会进一步优化，以提高功效和类似药物样的活性。一支由两名主要研究人员和几位合作者组成的多学科团队，建立了LRRK2病理学，秀丽隐杆线虫生物学和药物发现方面的专业知识。结果，已经生成了初步数据，以提供证据表明我们拟议的调查基于合理的科学原理。我们认为，我们提出的研究可能会促进针对LRRK2的潜在候选药物的发现，并且在哺乳动物模型中进一步验证可能会导致用于治疗和预防PD的新疗法。公共卫生相关性：帕金森病是一种毁灭性和致命的脑部疾病，影响了我们许多人。当前的PD治疗选择非常有限，没有治愈该疾病的治疗方法。由于LRRK2突变是家族性和零星PD的最常见原因，因此基于LRRK2的药物可能在受益于所有PD患者的情况下产生非常广泛的影响。我们提出的研究的意义将是发现用于治疗和预防PD的新药物铅的可能性。