Hyperphosphorylated tau aggregation-based Alzheimer’s disease early drug discovery

基于过度磷酸化 tau 聚集的阿尔茨海默病早期药物发现

• 批准号: 9904313
• 负责人: Min-Hao Kuo
• 金额: $ 43.67万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2022-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9904313
• 关键词: Abeta synthesis; Affect; Age; Alzheimer disease prevention; Alzheimer' s Disease; Alzheimer' s disease patient; Alzheimer' s disease therapeutic; American; Amyloid beta-Protein; Antibodies; Apomorphine; Apoptosis; Autopsy; Back; Biochemical; Biological Assay; Caregivers; Caring; Cell Line; Cells; Chemicals; Clinical; Clinical Trials; Collection; Development; Diffuse; Disease; Drug Design; Drug Targeting; Eligibility Determination; Environment; Escherichia coli; Failure; Fruit; Heparin; Hour; Impaired cognition; In Vitro; Investigational Drugs; Lead; Measures; Medical; Nerve Degeneration; Neurodegenerative Disorders; Neurofibrillary Tangles; Neurologic; Oxidation-Reduction; Parkinson Disease; Pathologic; Pharmaceutical Preparations; Pharmacologic Substance; Pilot Projects; Population; Prevention; Protocols documentation; Route; Senile Plaques; Solid; Technology; Variant; Work; base; cost; cytotoxic; cytotoxicity; disorder control; drug development; drug discovery; follow-up; high throughput screening; hyperphosphorylated tau; inhibitor/antagonist; innovation; loved ones; neuroblastoma cell; neurofibrillary tangle formation; novel; prevent; programs; screening; small molecule; socioeconomics; tau Proteins; tau aggregation; therapeutic development; wasting

Hyperphosphorylated tau aggregation-based Alzheimer’s disease early drug discovery Alzheimer’s disease (AD) is an irreversible neurodegenerative disease affecting 47 million people worldwide, and costs 604 billion US dollars every year for medical expenses. To date, there is no cure or prevention for AD. The two defining features of AD are Aβ plaques and neurofibrillary tangles that are composed of hyperphosphorylated tau. While the AD drug discovery landscape has been dominated by anti-Aβ measures, recurring failures of clinical trials argue strongly that a realignment of the drug target and strategies is needed to make a breakthrough in AD therapeutics development. Indeed, multiple lines of evidence suggest that the pre- tangle stage of hyperphosphorylated tau aggregates cause diffusible cytotoxicity that likely underlies neurodegeneration. Screening for compounds that prevent hyperphosphorylated tau from forming the cytotoxic aggregates thus affords a more viable route for AD drug discovery. The tangle-centric drug design has not come to fruition. A wide spectrum of compounds have been identified in multiple screens as inhibitors of tangle formation, but later found to be false-positive. One shared issue for these screens is the use of an unmodified tau protein that requires an inducer, e.g., heparin, for efficient aggregation in a reducing environment. The assay subject (tau) lacks the pathological mark of hyperphosphorylation, and the disease relevance of the inducer has not been substantiated. To overcome these hurdles, we have developed the PIMAX technology that produces hyperphosphorylated tau (p-tau) in E. coli. Purified p-tau fibrillizes autonomously (without an inducer), and causes apoptosis of different cells including a neuroblastoma cell line. This inducer-free, p-tau aggregation assay has a Z’ value of 0.699, and a coefficient of variation (CV) of 8.3%. These parameters qualify our p-tau aggregation assay as a robust HTS platform. Using this assay, we conducted a pilot screen for 1,280 compounds for their ability to modulate the aggregation of p- tau. We then used novel biochemical and cell-based secondary assays to verify the candidate compounds. We found that an active neurological drug is a potent p-tau aggregation inhibitor, which also protects cells from p- tau cytotoxicity. These preliminary studies afford solid evidence for the values of p-tau in the quest for AD therapeutics. This R01 project is the very first that uses the pathophysiologically relevant hyperphosphorylated tau for AD drug discovery. We will follow our pilot screen protocols to conduct a high-throughput screen of 100,000 compounds for their ability to inhibit p-tau aggregation and to protect cells. This early discovery project will conclude with two major products: (1) A collection of chemical hits with confirmed p-tau aggregation inhibitory and cytoprotective activities, and (2) a follow-up proposal with comprehensive plans to identify the optimal chemical lead for AD drug development.

基于过度磷酸化tau聚集的阿尔茨海默病早期药物发现 阿尔茨海默病（AD）是一种不可逆的神经退行性疾病，影响全球4700万人， 每年的医疗费用高达六千零四十亿美元。到目前为止，还没有治愈或预防 AD. AD的两个定义特征是Aβ斑块和神经元缠结， 过度磷酸化的tau蛋白虽然AD药物发现领域一直由抗A β措施主导， 临床试验的反复失败有力地表明，需要重新调整药物靶点和策略， 在AD治疗药物开发方面取得突破。事实上，多条证据表明， 过度磷酸化tau聚集体缠结阶段导致可能导致 神经变性筛选防止过度磷酸化的tau形成细胞毒性的化合物 因此，聚集体为AD药物发现提供了更可行的途径。 以缠结为中心的药物设计尚未实现。广泛的化合物已经被 在多次筛选中被鉴定为缠结形成的抑制剂，但后来发现是假阳性。一个共享 这些筛选的问题是使用需要诱导剂的未修饰的tau蛋白，例如，肝素，用于有效 在还原环境中聚集。测定受试者（tau）缺乏以下病理标记： 过度磷酸化，并且诱导物的疾病相关性尚未得到证实。克服这些 我们已经开发了PIMAX技术，在E.杆菌 纯化的p-tau自主地（在没有诱导剂的情况下）灭活，并引起不同细胞的凋亡，包括 神经母细胞瘤细胞系该无诱导剂的p-tau聚集测定具有0.699的Z'值和0.699的系数。 变异系数（CV）为8.3%。这些参数使我们的p-tau聚集测定法成为稳健的HTS平台。使用 在该测定中，我们对1，280种化合物进行了初步筛选，以确定它们调节β- τ的然后，我们使用新的生物化学和基于细胞的二级检测来验证候选化合物。我们 发现一种活跃的神经药物是一种有效的p-tau聚集抑制剂，它也可以保护细胞免受p-tau聚集的影响。 tau细胞毒性。这些初步研究为p-tau蛋白在寻找AD中的价值提供了坚实的证据 治疗学这个R 01项目是第一个使用病理生理学相关的过度磷酸化的 用于AD药物发现。我们将按照我们的试点筛选方案进行高通量筛选， 100，000种化合物，它们能够抑制p-tau聚集并保护细胞。这个早期发现项目 将得出两个主要产物：（1）具有确认的p-tau聚集抑制性的化学命中的集合 和细胞保护活性，以及（2）后续建议与全面计划，以确定最佳的 AD药物开发的化学先导。