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

描述(由申请人提供)：用于阿尔茨海默病药物筛选的高通量Tau齐聚物分析项目摘要：治疗阿尔茨海默病的疾病修饰药物有大量且快速增长的未得到满足的需求。目前，全世界有1800万例AD病例；到2025年，这一数字预计将增加到3400万例。目前，只有5种温和有效的治疗AD症状的药物存在，但没有一种药物治疗潜在的神经退行性变过程。随着人们对tau在神经退行性变中的作用越来越了解，tau正在成为疾病修改药物(DMD)开发的一个更突出的目标。Tau蛋白MAPT基因的突变是痴呆的原因，tau病理与AD的进展密切相关。与此同时，基于淀粉样蛋白假说的晚期临床治疗失败提出了仅以A2为靶点的问题。已经出现了强有力的证据，表明tau寡聚体在AD和20多种其他神经退行性疾病的疾病发病机制中发挥直接作用(Brunden等人。2008年；Davidowitz等人。2008年)。为了发现靶向tau的药物，开发了一些方法来选择抑制tau自身相互作用的化合物，并选择了一种使用AlphaScreen检测技术的检测方法来进一步开发高通量筛选(Chatterjee等人。2008年)。此外，Sierks博士(亚利桑那州立大学)开发的噬菌体展示-原子力显微镜方法被用来分离与tau寡聚体特异结合的抗体片段(ScFv)，这些抗体片段将被用于基于细胞的筛选分析。拟议计划的具体目标如下：“将tau寡聚体分析转换为HTS格式”将tau寡聚体分析转换为HTS格式“将tau寡聚体分析转移到密歇根高通量筛选中心(MHTSC)，用于高度优化的化合物库(100,000种化合物)的自动化和筛选，并进行药物化学分析以模拟药效团并选择额外的化学类型进行筛选”在基于细胞的分析中使用tau寡聚体特异性抗体片段选择化合物建议的第二阶段计划的预期结果是选择至少三个或更多针对tau寡聚体的先导候选化合物，这些化合物将在第三阶段计划期间开发并在AD和tauopathies动物模型中进行评估。为了实现这一结果，高通量分析将被优化并转移到密歇根高通量筛选中心，在那里，他们将在Robert Kilkuskie Hits博士的指导下对大约100,000种化合物的选择集库进行筛选，并将开发一个结构性药效团模型。有毒化合物将通过神经细胞毒性测试来消除。将选择三个或更多具有高灵敏度和特异性的单链抗体，用于通过基于细胞的体外分析来鉴定抑制tau自结合的化合物。抗体片段除了实现主要目标外，还支持该公司的其他计划，包括其tau生物标记物开发计划和tau免疫治疗计划。然而，该计划的主要目标是推进tau寡聚体药物发现平台，以确定活性抑制剂作为IND使能研究的主要候选药物。关键词阿尔茨海默病，肌萎缩侧索硬化症，神经退行性疾病，药物发现，tau寡聚体，噬菌体展示，抗体片段，单链抗体，高通量筛选 公共卫生相关性：拟议的计划重点是开发一种针对tau寡聚体的高通量筛查试验，用于治疗阿尔茨海默病(AD)的药物发现。该项目的灵感来自于观察到，tau寡聚体的积累已被证明与阿尔茨海默病和自闭症小鼠模型中的神经元丢失和记忆障碍有很好的相关性。OLIGOMERIX已经开发出体外试验来筛选抑制细胞毒性tau寡聚体形成的化合物。本程序的目标是1)。将tau齐聚物分析转换为HTS格式；2)将测试转移到密歇根高通量筛选中心(MHTSC)，以实现高度优化的化合物库(100,000化合物)的自动化和筛选，并进行药物化学分析以模拟药效团；使用tau寡聚体特异性抗体片段和基于细胞的筛选来选择化合物，以确定未来动物研究和临床前开发的主要候选者。

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.