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

基本信息

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
项目类别：
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&apos 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

项目摘要

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）：阿尔茨海默氏病（AD）是老年人痴呆症的主要原因，美国发病率为540万，每年成本为1830亿美元。没有有效的治疗方法。该建议是对NIA要求R21高风险/高奖励提案的要求，以生成新的AD药物发现策略。它的重点是当前的知识，表明AD中的显着神经元损害可以归因于有毒AB低聚物的影响。这些微小的扩散分子与淀粉样蛋白斑块不同，被认为可以通过靶向突触和破坏突触可塑性的能力来激发AD记忆丧失。当前的努力阻止AD的努力主要关注代谢抑制剂，这些抑制剂阻断了AB单体的积累以及去除单体或其毒性组件的被动疫苗。这些努力尚未成功。该提案引入了一种替代策略，该策略重点介绍了低聚物毒性机理的第一步。为了使弹性损害，低聚物必须首先与细胞受体结合。这些受体介导了低聚物与特定神经元的缔合，并结合将其转换为突触毒素反应。尽管受体身份对于阐明毒性机制很有价值，但即使没有这种知识，也可以使用将低聚物受体结合作为药物发现的靶标也是可行的。所需的是一种适用于结合拮抗剂的大型高通量筛选（HTS）的测定。我们提出了一种新的策略，使用一种产生人工纳米级膜（纳米尺度）的方法来实现这一目标。这是一种完善的方法，在这里已改编，以提供突触质膜蛋白的无偏和可溶性制剂。已经证明了低聚物与突触质膜（SPM）纳米盘的结合，并表现出了配体 - 受体相互作用的期望的特征。结合反应已适应了均匀的化学发光测定法，非常适合HTS，用于低聚物结合的拮抗剂。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主要筛选非常大的化合物库所需的带宽和精度。与该项目分开的这些SPM纳米盘的平行研究有望鉴定受体蛋白。我们的药物发现方法遵循筛选树，在基于细胞的结合和毒性的基于细胞的测定中，使用SPM-Nanodiscs从主要测定中进行了命中，首先是合成的低聚物，然后是脑源性的低聚物。纳米轴HTS和辅助屏幕将首先使用一个小库进行优化，然后大大扩展。热门资源o西北分子创新和药物发现中心将根据需要实施。预计将在两年结束前实现的目标是为NIH的主持下现在可用的大型图书馆建立一个功能齐全的策略。结果最终具有强大的潜力，可以发现靶向AD发病机理的不流失但重要方面的铅化合物。