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）：阿尔茨海默氏病 (AD) 是导致老年人痴呆的主要原因，在美国发病率为 540 万，每年造成的费用为 1,830 亿美元。没有有效的治疗方法。该提案是为了响应 NIA 对 R21 高风险/高回报提案的请求，以生成新的 AD 药物发现策略。它重点关注当前的知识，表明 AD 中的显着神经元损伤可归因于有毒抗体寡聚物的影响。这些可扩散的小分子与淀粉样斑块不同，被认为通过其靶向突触和破坏突触可塑性的能力而引发 AD 记忆丧失。目前预防 AD 的努力主要集中在阻止抗体单体积累的代谢抑制剂和消除单体或其有毒组装体的被动疫苗上。这些努力尚未取得成功。该提案引入了一种替代策略，重点关注低聚物毒性机制的第一步。为了引起损伤，寡聚体必须首先与细胞受体结合。这些受体介导寡聚体与特定神经元的结合，并将结合转导为突触毒性反应。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点也是可行的。所需要的是适合于结合拮抗剂的大规模高通量筛选(HTS)的测定法。我们提出了一种新的策略来实现这一目标，使用一种生成人造纳米级膜（Nanodiscs）的方法。这是一种行之有效的方法，已在此进行了调整，以提供突触质膜蛋白的公正且功能性的可溶性制剂。寡聚物与突触质膜 (SPM) 纳米圆盘的结合已得到证实，并表现出配体-受体相互作用的预期特征。结合反应已适应均相化学发光测定，非常适合低聚物结合拮抗剂的 HTS。与高含量、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库进行初步筛选所需的带宽和精度。独立于该项目的这些 SPM-Nanodisc 的并行研究预计将鉴定受体蛋白。我们的药物发现方法遵循筛选树，其中使用 SPM-Nanodiscs 的初级测定中的命中在基于细胞的结合和毒性测定中进行验证，首先使用合成寡聚物，然后使用脑源性寡聚物。 Nanodisc HTS 和二级屏幕将首先使用小型库进行优化，然后进行大幅扩展。西北分子创新和药物发现中心的热门资源将根据需要实施。该目标预计将在两年内实现，即为 NIH 赞助下的超大型图书馆建立一个功能齐全的 HTS 战略。结果最终具有发现针对 AD 发病机制中尚未充分开发但重要的方面的先导化合物的巨大潜力。 公共健康相关性：该提案是为了响应 NIA 对 R21 高风险/高回报提案的请求，以产生新的阿尔茨海默病 (AD) 药物发现策略。 AD 是老年人痴呆症的主要原因，美国的发病率为 540 万，每年造成的损失达 1,830 亿美元。没有有效的治疗方法。该项目预计将具有巨大的潜力，能够识别出以突触毒性抗体寡聚物与其受体结合为目标的先导化合物，这是 AD 发病机制中尚未充分利用但至关重要的早期步骤。