High throughput tau oligomer assay for drug screening for Alzheimer's disease

高通量 tau 寡聚体检测用于阿尔茨海默病药物筛选

• 批准号: 8121384
• 负责人: JAMES G. MOE
• 金额: $ 73.22万
• 依托单位: OLIGOMERIX, INC
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2013-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8121384
• 关键词: 2-cyclopentyl-5-(5-isoquinolylsulfonyl)-6-nitro-1H-benzo(D)imidazole; Adopted; Algorithms; Alzheimer' s Disease; Amyloid; Animal Model; Animal Testing; Animals; Atomic Force Microscopy; Automation; Binding; Biological Assay; Biological Markers; Brain; Cell model; Cells; Characteristics; Chemistry; Clinical; Dementia; Detection; Development; Disease; Disease Progression; Dose; Dot Immunoblotting; Drug Delivery Systems; Enzyme-Linked Immunosorbent Assay; Failure; Future; Genes; Goals; Health; Immunoglobulin Fragments; Immunotherapeutic agent; In Vitro; Inhibitory Concentration 50; Lead; Libraries; Measures; Memory impairment; Methods; Michigan; Modeling; Mutation; Nerve Degeneration; Neurodegenerative Disorders; Outcome; Pathogenesis; Pathology; Performance; Phage Display; Pharmaceutical Chemistry; Pharmaceutical Preparations; Phase; Play; Poison; Preclinical Drug Evaluation; Process; Program Development; Promega; Property; Protocols documentation; Reading; Reproducibility; Role; Sampling; Screening procedure; Sensitivity and Specificity; Signal Transduction; Specificity; Staging; Symptoms; Tauopathies; Technology; Therapeutic; Time; Toxic effect; base; cytotoxic; dimer; drug discovery; high throughput screening; improved; in vitro Assay; in vivo; inhibitor/antagonist; monomer; mouse model; neuron loss; pharmacophore; preclinical study; programs; response; success; task analysis; tau Proteins; tau aggregation; tool

DESCRIPTION (provided by applicant): High Throughput Tau Oligomer Assay for Drug Screening for Alzheimer's Disease Project Summary: There is a large and rapidly growing unmet need for disease modifying drugs for Alzheimer's disease. Currently there are 18 million cases of AD worldwide; by 2025 this number is expected to increase to 34 million. Presently, only 5 mildly effective AD symptom-treating drugs exist, but none that treat the underlying neurodegenerative processes. Tau is becoming a more prominent target for the development of disease- modifying drugs (DMDs), as its role in neurodegeneration is becoming better understood. Mutations in the gene for tau protein MAPT are causative of dementia and tau pathology correlates well with AD progression. At the same time, late stage clinical failures for therapeutics based on the amyloid hypothesis have raised questions on solely targeting A2. Strong evidence has emerged implicating tau oligomers as playing a direct role in disease pathogenesis for AD and over 20 other neurodegenerative diseases (Brunden et al. 2008; Davidowitz et al. 2008). To discover drugs targeting tau oligomerization methods were developed to select compounds inhibiting tau self-interaction, and an assay using AlphaScreen detection technology was selected for further development for high throughput screening (Chatterjee et al. 2008). In addition, the phage display- atomic force microscopy method developed by Dr. Sierks (ASU) was used to isolate antibody fragments (scFvs) specifically binding to tau oligomers that will be adopted in the cell based screening assays. The specific aims of the proposed program are as follows: " Convert the tau oligomer assay to HTS format " Transfer Assay to the Michigan High Throughput Screening Center (MHTSC) for automation and screening of a highly optimized compound library (100,000 compounds) and carry out medicinal chemistry analysis to model the pharmacophore and select additional chemotypes for screening " Select compounds using tau oligomer specific antibody fragments in cell based assays The anticipated outcome of the proposed Phase II program is the selection of at least three or more lead candidate compounds targeting tau oligomers that will be developed during the Phase III program and evaluated in animal models of AD and tauopathies. To attain this result, the high throughput assay will be optimized and transferred to the Michigan High Throughput Screening Center where their Select Set library of approximately 100,000 compounds will be screened under the direction Dr. Robert Kilkuskie Hits will be validated and a structural pharmacaophore model will be developed. Toxic compounds will be eliminated using a neurocytotoxicity assay. Three or more scFvs will be selected with high sensitivity and specificity that will be used to identify compounds inhibiting tau self-association using in vitro cell based assays. Antibody fragments, in addition to enabling the primary goal, are supportive of the company's other programs including its tau biomarker development program and its tau immunotherapeutic program. However, the primary goal of the program is to advance the tau oligomer drug discovery platform to identify active inhibitors as lead candidates for IND enabling studies. Key Words Alzheimer's disease, tauopathy, neurodegenerative disease, drug discovery, tau oligomer, phage display, antibody fragment, scFv, high throughput screening PUBLIC HEALTH RELEVANCE: The proposed program focuses on developing a high throughput screening assay targeting tau oligomers for drug discovery for Alzheimer's disease (AD). This project was inspired by observations that accumulation of tau oligomers has been shown to correlate well with neuronal loss and memory impairment in AD and in tauopathy mouse models. OLIGOMERIX has developed in vitro assays for screening compounds that inhibit the formation of cytotoxic tau oligomers. This program aims to 1.) .Convert the tau oligomer assay to HTS format; 2.) Transfer Assay to the Michigan High Throughput Screening Center (MHTSC) for automation and screening of a highly optimized compound library (100,000 compounds) and carry out medicinal chemistry analysis to model the pharmacophore; 3.) Select compounds using tau oligomer specific antibody fragments and cell based screening to identify lead candidates for future animal studies and pre-clinical development.

描述（由申请人提供）：高通量TAU低聚物分析用于筛查阿尔茨海默氏病的药物筛查项目摘要：对于阿尔茨海默氏病的疾病修改药物的巨大而迅速增长的未满足的需求。目前，全球有1800万例广告案例；到2025年，这一数字预计将增加到3400万。目前，只有5种轻度有效的AD症状治疗药物，但没有治疗潜在的神经退行性过程。 Tau正在成为疾病改变药物（DMD）的更为突出的靶标，因为它在神经退行性中的作用正在变得更好地理解。 Tau蛋白MAPT基因中的突变是痴呆症的原因，而Tau病理学与AD进展很好地相关。同时，基于淀粉样假说的治疗剂的后期临床失败提出了有关仅针对A2的问题。有力的证据表明，tau低聚物是在AD和其他20种其他神经退行性疾病中发挥直接作用（Brunden等人，2008； Davidowitz等，2008）。为了发现靶向tau寡聚方法的药物以选择抑制tau自我相互作用的化合物，并选择了使用Alphascreen检测技术进行测定以进行进一步开发以进行高吞吐量筛选（Chatterjee等，2008）。此外，由Sierks博士（ASU）开发的噬菌体显示 - 原子力显微镜方法用于分离抗体片段（SCFV），该抗体片段（SCFV）特异性结合了与Tau低聚物的结合，这些抗体片段将在基于细胞的筛选测定中采用。 The specific aims of the proposed program are as follows: " Convert the tau oligomer assay to HTS format " Transfer Assay to the Michigan High Throughput Screening Center (MHTSC) for automation and screening of a highly optimized compound library (100,000 compounds) and carry out medicinal chemistry analysis to model the pharmacophore and select additional chemotypes for screening " Select compounds using tau oligomer specific antibody fragments in基于细胞的测定，提议的II期计划的预期结果是针对靶向TAU低聚物的至少三个或更多的铅候选化合物，这些化合物将在III期计划期间开发，并在AD和Tauopathies的动物模型中进行评估，以实现该结果，以实现高吞吐量的范围。 Kilkuskie的热门歌曲将得到验证，并将开发结构性的药物模型。使用神经细胞毒性测定法将消除有毒化合物。将选择具有高灵敏度和特异性的三个或更多SCFV，这些SCFV将用于识别使用基于体外细胞的测定法抑制TAU自我关联的化合物。除了实现主要目标外，抗体碎片还支持该公司的其他计划，包括其TAU生物标志物开发计划和TAU免疫治疗计划。但是，该计划的主要目标是促进TAU低聚物药物发现平台，以鉴定活跃的抑制剂作为IND启用研究的主要候选者。关键词阿尔茨海默氏病，陶氏病，神经退行性疾病，药物发现，tau低聚物，噬菌体显示，抗体片段，SCFV，高吞吐量筛查 公共卫生相关性：拟议的计划着重于开发针对阿尔茨海默氏病（AD）药物发现的高吞吐量筛查测定法。该项目的灵感来自观察到，已经证明TAU低聚物的积累与AD和Tauopathy小鼠模型中的神经元损失和记忆障碍良好相关。寡聚物已开发出用于筛选化合物的体外测定，从而抑制了细胞毒性tau低聚物的形成。该程序的目的是1。）。将tau低聚物测定法转化为HTS格式； 2.）将测定法转移到密歇根州高吞吐量筛查中心（MHTSC），以进行自动化和筛选高度优化的化合物库（100,000种化合物），并进行药物化学分析，以模拟药物学； 3.）选择使用Tau低聚物特异性抗体片段和基于细胞的筛查的化合物，以鉴定未来动物研究和临床前发育的铅候选者。