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）：我们申请的总体目标是将有关 PD 发病机制关键角色 LRRK2 的最新研究成果转化为新型 PD 疗法的药物发现管道。富含亮氨酸重复激酶 2 (LRRK2) 的突变是迄今为止在家族性和散发性 PD 中发现的最常见原因。由于突变的 LRRK2 蛋白会导致 LRRK2 在体外过度激活并在神经元中产生细胞毒性，因此针对 LRRK2 的治疗可能对 PD 非常有效。我们还认为 LRRK2 代表了治疗开发的难得机会，因为它包含两个可成药的酶模块（激酶结构域和 GTP 酶结构域），可能提供多个药物作用位点。我们建议采用两种互补的方法来开发针对 LRRK2 的 PD 药物。具体目标 1 是使用重组全长 LRRK2 对现有药物以及激酶抑制剂和 GTP 结合阻断剂的其他化学文库进行高通量筛选，这些药物将通过不同的作用机制阻止 LRRK2 激活。具体目标 2 是利用新开发的秀丽隐杆线虫 PD 模型进行 LRRK2 抑制剂的体内测定。最近的研究表明，线虫可以作为早期药物筛选和验证的强大整体动物模型。我们最近建立了表达突变体 LRRK2 的转基因线虫动物，其表现出与 PD 相关的神经变性和行为障碍的易于量化的表型。这种 LRRK2 连接 PD 的线虫模型将用于验证和表征体外化学筛选中有前景的 LRRK2 抑制化合物。由此产生的 LRRK2 抑制剂候选物可能会进一步优化，以获得更好的功效和类似药物的活性。一个由两名主要研究人员和几名在 LRRK2 病理学、线虫生物学和药物发现方面具有专业知识的合作者组成的多学科团队已经成立。因此，已经生成了初步数据，以证明我们提出的调查是基于合理的科学原理。我们相信，我们提出的研究可能会促进发现针对 LRRK2 的潜在候选药物，并且在哺乳动物模型中进一步验证后可能会产生治疗和预防 PD 的新疗法。公共卫生相关性：帕金森病是一种破坏性且致命的脑部疾病，影响许多美国人。目前帕金森病的治疗选择非常有限，并且无法治愈该疾病。由于 LRRK2 突变是迄今为止家族性和散发性 PD 的最常见原因，因此基于 LRRK2 的药物可能会产生非常广泛的影响，使所有 PD 患者受益。我们提出的研究的意义在于有可能发现治疗和预防帕金森病的新药物先导物。