Alzheimer's Drug Discovery Using Unique Nanotechnology Platform

使用独特的纳米技术平台发现阿尔茨海默病药物

• 批准号: 8446087
• 负责人: WILLIAM L KLEIN
• 金额: $ 24.74万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-15 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8446087
• 关键词: Affinity; Alzheimer' s Disease; Artificial Membranes; Binding; Biochemical; Biological Assay; Brain; Cell membrane; Cells; Characteristics; Chemiluminescence assay; Collection; Coupled; Data; Dementia; Elderly; Exhibits; Funding; Goals; Incidence; Investigation; Knowledge; Lead; Libraries; Ligands; Mediating; Membrane; Membrane Proteins; Memory Loss; Metabolic; Methodology; Molecular; Molecular Conformation; Nanotechnology; Neurons; Pathogenesis; Preparation; Protein Binding; Proteins; Publishing; Reaction; Recombinant Proteins; Recombinants; Research; Research Personnel; Resources; Rest; Screening procedure; Senile Plaques; Structure; Synapses; Synaptic Receptors; Synaptic plasticity; Technology; Toxic effect; Trees; United States National Institutes of Health; Vaccines; Validation; base; cost; drug discovery; effective therapy; gain of function; high reward; high risk; high throughput screening; inhibitor/antagonist; innovation; innovative technologies; monomer; nanodisk; nanoscale; novel; novel strategies; prevent; receptor; receptor binding; response

DESCRIPTION (provided by applicant): Alzheimer's disease (AD) is the major cause of dementia in the elderly, with a US incidence of 5.4 million and an annual cost of $183 billion. There is no effective treatment. This proposal is in response to an NIA request for R21 high risk/high reward proposals to generate new AD drug discovery strategies. It focuses on current knowledge indicating that significant neuron damage in AD can be attributed to the impact of toxic Ab oligomers. These small diffusible molecules are distinct from amyloid plaques and are thought to instigate AD memory loss through their ability to target synapses and disrupt synaptic plasticity. Current efforts to prevent AD largely focus on metabolic inhibitors that block accumulation of the Ab monomer and on passive vaccines that remove either the monomer or its toxic assemblies. These efforts have not as yet been successful. This proposal introduces an alternative strategy that focuses on the first step in the mechanism of oligomer toxicity. To elici damage, oligomers must first bind to cellular receptors. These receptors mediate association of oligomers with particular neurons and transduce binding into synaptotoxic responses. Although receptor identity would be valuable for elucidating the mechanism of toxicity, it is feasible even without this knowledge to use oligomer-receptor binding as a target for drug discovery. What is required is an assay suitable for large- scale high-throughput screening (HTS) of binding antagonists. We propose a novel strategy to achieve this goal using an approach that generates artificial nanoscale membranes (Nanodiscs). This is a well-established methodology that has been adapted here to provide unbiased and functional soluble preparations of synaptic plasma membrane proteins. Binding of oligomers to synaptic plasma membrane (SPM) Nanodiscs has been demonstrated and exhibits characteristics expected of ligand-receptor interaction. The binding reaction has been adapted to a homogeneous chemiluminescence assay well-suited to HTS for antagonists of oligomer binding. Unlike high content, cell-based assays, the biochemical assay for binding to soluble receptors has the bandwidth and precision required for the primary screening of very large libraries of compounds. Parallel investigations of these SPM-Nanodiscs, separate from this project, are expected to identify the receptor protein(s). Our Approach to drug discovery follows a screening tree in which hits from the primary assay using SPM-Nanodiscs are validated in cell-based assays for binding and toxicity, first with synthetic oligomers and then with brain-derived oligomers. The Nanodisc HTS and secondary screens will be optimized at first using a small library and then greatly expanded. Hit-to-lead resources o the Northwestern Center for Molecular Innovation and Drug Discovery will be implemented as needed. The Aim, expected to be achieved by the end of two years, is to establish a fully functioning strategy for HTS of the very large libraries now available under the auspices of NIH. Results ultimately have strong potential for discovering lead compounds that target an underexploited but significant aspect of AD pathogenesis. PUBLIC HEALTH RELEVANCE: This proposal is in response to an NIA request for R21 high risk/high reward proposals to generate new Alzheimer's disease (AD) drug discovery strategies. AD is the major cause of dementia in the elderly, with a US incidence of 5.4 million and an annual cost of $183 billion. There is no effective treatment. This project is expected to have strong potential to identify lead compounds that target binding of synaptotoxic Ab oligomers to their receptors, an underexploited but critical early-step in AD pathogenesis.

描述（由申请人提供）：阿尔茨海默病（AD）是老年人痴呆的主要原因，在美国的发病率为540万，每年的成本为1830亿美元。没有有效的治疗方法。本提案是为了响应NIA对R21高风险/高回报提案的请求，以产生新的AD药物发现策略。它侧重于目前的知识表明，AD中显著的神经元损伤可归因于毒性Ab寡聚物的影响。这些小的可扩散分子不同于淀粉样斑块，并且被认为通过它们靶向突触和破坏突触可塑性的能力来引发AD记忆丧失。目前预防阿尔茨海默病的努力主要集中在阻止Ab单体积累的代谢抑制剂和去除单体或其毒性组装的被动疫苗上。这些努力尚未取得成功。本提案介绍了一种替代策略，重点关注低聚物毒性机制的第一步。为了引起损伤，低聚物必须首先与细胞受体结合。这些受体介导寡聚物与特定神经元的结合，并将结合转化为突触毒性反应。尽管受体身份对于阐明毒性机制很有价值，但即使没有这些知识，使用寡聚物受体结合作为药物发现的靶标也是可行的。所需要的是一种适合大规模高通量筛选（HTS）结合拮抗剂的检测方法。我们提出了一种新的策略来实现这一目标，使用一种产生人工纳米级膜（纳米片）的方法。这是一种完善的方法，在这里已经适应了提供公正和功能可溶的突触质膜蛋白制备。低聚物与突触质膜（SPM）纳米盘的结合已被证实，并表现出配体-受体相互作用的特征。结合反应已适应于均匀的化学发光测定，非常适合于HTS低聚物结合拮抗剂。与高含量的、基于细胞的测定不同，与可溶性受体结合的生化测定具有对非常大的化合物文库进行初步筛选所需的带宽和精度。这些spm纳米圆盘的平行研究，独立于本项目，有望鉴定受体蛋白。我们的药物发现方法遵循筛选树，在筛选树中，使用spm -纳米圆盘进行的初级分析命中的结果在基于细胞的分析中进行结合和毒性验证，首先使用合成低聚物，然后使用脑源低聚物。Nanodisc HTS和二级屏幕将首先使用一个小库进行优化，然后大大扩展。根据需要，西北分子创新和药物发现中心的“从成功到领先”资源将得到实施。该目标预计将在两年内实现，目的是为目前由NIH赞助的大型图书馆的HTS建立一个全面运作的战略。结果最终具有发现针对阿尔茨海默病发病机制中尚未开发但重要方面的先导化合物的强大潜力。