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: 8548221
• 负责人: WILLIAM L KLEIN
• 金额: $ 18.44万
• 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/8548221
• 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; 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; public health relevance; receptor; receptor binding; response; screening

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.

描述（由申请人提供）：阿尔茨海默病（AD）是老年痴呆症的主要原因，在美国的发病率为540万，每年的费用为1830亿美元。没有有效的治疗方法。该提案是为了响应NIA对R21高风险/高回报提案的要求，以产生新的AD药物发现策略。它侧重于目前的知识，表明AD中的显著神经元损伤可归因于有毒Ab寡聚体的影响。这些小的可扩散分子与淀粉样蛋白斑块不同，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AD的努力主要集中在阻断Ab单体积累的代谢抑制剂和去除单体或其毒性组装体的被动疫苗上。这些努力尚未取得成功。该提案介绍了一种替代策略，其重点是低聚物毒性机制的第一步。为了引发损伤，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，并介导突触毒性反应。虽然受体身份对于阐明毒性机制是有价值的，但即使没有这种知识，使用寡聚体-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。我们提出了一种新的策略来实现这一目标，使用的方法，产生人工纳米级膜（纳米盘）。这是一个完善的方法，已在这里进行了调整，以提供无偏见的和功能性的突触质膜蛋白的可溶性制剂。寡聚体与突触质膜（SPM）纳米盘的结合已经被证明，并且表现出配体-受体相互作用的预期特征。结合反应已适应于均相化学发光测定，非常适合HTS的寡聚体结合的拮抗剂。与高含量的基于细胞的测定不同，用于结合可溶性受体的生物化学测定具有初步筛选非常大的化合物库所需的带宽和精度。与本项目分开的这些SPM纳米盘的平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，其中在基于细胞的结合和毒性测定中验证使用SPM纳米盘的初步测定的命中，首先使用合成寡聚体，然后使用脑源性寡聚体。Nanodisc HTS和二级筛选将首先使用小型文库进行优化，然后进行大幅扩展。将根据需要实施西北分子创新和药物发现中心的命中领先资源。目标，预计将在两年内实现，是建立一个全面运作的战略，HTS的非常大的图书馆，现在可以在美国国立卫生研究院的主持下。结果最终有很大的潜力，发现铅化合物的目标是开发不足，但重要的方面，AD的发病机制。