Screening for enhancers of sAPPalpha

筛选 sAPPalpha 增强子

• 批准号: 9265756
• 负责人: Varghese John
• 金额: $ 19.25万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-01 至 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9265756
• 关键词: Abeta clearance; Affinity; Aging; Agonist; Alzheimer' s Disease; Alzheimer' s disease model; Amyloid beta-Protein; Amyloid beta-Protein Precursor; Animal Model; Artificial Membranes; Australia; Biological Assay; Blood - brain barrier anatomy; Brain; Brain Diseases; Cell Line; Cell Membrane Permeability; Cells; Chemicals; Cleaved cell; Clinic; Clinical; Clinical Trials; Cognition; Cognitive; Computer Simulation; Country; Data; Dementia; Development; Disease; Disease Progression; Dose; Down-Regulation; Drug Kinetics; Drug usage; Embryo; Enhancers; Environment; Etiology; Evaluation; Event; Foundations; Gel; Generations; Genetic Screening; Goals; Hippocampus (Brain); Human; Human Amyloid Precursor Protein; Impaired cognition; In Vitro; Infusion procedures; Lead; Libraries; Mediating; Membrane; Memory; Molecular; Mus; N-Methyl-D-Aspartate Receptors; Nausea and Vomiting; Neuroblastoma; Neurodegenerative Disorders; Neurons; Oral; Outcome; Pathogenicity; Pathologic; Pathway interactions; Patients; Penetrance; Peptides; Permeability; Pharmaceutical Preparations; Postoperative Nausea and Vomiting; Postoperative Period; Probability; Process; Production; Rattus; Resource Sharing; Role; Senile Plaques; Slice; Synapses; Testing; Therapeutic; Tissues; Treatment Efficacy; Tropisetron; Validation; alpha secretase; amyloid precursor protein processing; base; beta secretase; bioprinting; brain tissue; chemical genetics; clinical candidate; clinical development; cost; density; excitotoxicity; gamma secretase; genetic approach; high throughput screening; improved; in vitro Model; in vivo; insight; memory retention; mild cognitive impairment; mouse model; neuroblastoma cell; novel; novel strategies; novel therapeutics; preclinical efficacy; prevent; public health relevance; receptor binding; response; screening; success; symptom treatment; targeted treatment; therapeutic development; therapeutic target

 DESCRIPTION (provided by applicant): Alzheimer's disease (AD) currently afflicts more than 5.4 million people in the US at an estimated cost greater than $200 billion per year. Currently approved drugs offer only short-term symptomatic relief but do not alter disease progression. The neuritic plaques that are a hallmark of AD brain result from increased generation and/or decreased clearance of the amyloid beta peptide (Aβ) which originates from amyloid precursor protein (APP). APP may be processed by two pathways. In one, it undergoes sequential β-secretase and γ-secretase cleavages to generate soluble amyloid precursor protein beta (sAPPβ) and Aβ; in the other, it is cleaved by an α-secretase to generate soluble amyloid precursor protein alpha (sAPPα), a pro-cognitive peptide that may prevent AD. This project proposes to advance our previous studies that successfully identified the sAPPα-enhancer tropisetron (F03) with the goal of identifying new sAPPα-enhancers as new chemical entities (NCEs) constituting a novel class of therapeutics for AD. Our strong foundation of preliminary data indicates a high probability that the specific aims of this project can be achieved. We have had early success utilizing our iterative, hierarchical chemical-genetics screening approach in the identification of F03 - a drug used for post- operative nausea and vomiting - from a small clinical compound library. F03 was shown to consistently but modestly increase sAPPα in vitro and in vivo in a mouse model of AD, and to significantly improve memory in the AD model after four weeks of oral treatment. F03 is now in clinical trials for mild cognitive impairment (MCI) due to AD as a repurposed drug. Here, we propose utilizing our high-throughput screening (HTS) assay, iterative screening flowscheme, and the large UCLA compound library to identify compounds with sAPPα-enhancing effects greater than F03. In the primary HTS screen, "hits" that increase sAPPα will be identified. In the secondary screen, hits are validated using a second in vitro model. Prioritized hits will then be further validated by ex vivo organotypic cultue of AD model brain and brain penetrance determined by parallel artificial membrane permeability analysis (PAMPA) and in vivo pharmacokinetic (PK) analysis. Finally, we will determine the mechanisms by which hits induce sAPPα enhancement using the CEREP Bioprint profile, drug affinity responsive target stability (DARTS), in silico target ID using the similarity ensemble approach, and by a new gel-enhanced target identification approach (GET). While other new approaches to identify therapeutics for AD and MCI are under development, their clinical outcomes remain uncertain. By identifying compounds with sAPPα enhancing effects greater than F03, we may find promising NCEs for further clinical development. Perhaps most importantly, our mechanistic studies of such enhancers may further elucidate the role of sAPPα in the etiology of AD, providing new direction for therapeutic development.

 描述（由申请人提供）：阿尔茨海默病 (AD) 目前在美国困扰着超过 540 万人，估计每年花费超过 2000 亿美元。目前批准的药物只能缓解短期症状，但不能改变疾病进展。神经炎斑块是 AD 大脑的一个标志，是由源自淀粉样前体蛋白 (APP) 的淀粉样β肽 (Aβ) 生成增加和/或清除减少所致。 APP 可以通过两种途径进行加工，一种是通过连续的 β 分泌酶和 γ 分泌酶裂解生成可溶性淀粉样前体蛋白 β (sAPPβ) 和 Aβ；另一种是通过 α 分泌酶裂解生成可溶性淀粉样蛋白前体蛋白 β。淀粉样前体蛋白 α (sAPPα)，一种可能预防 AD 的促认知肽。该项目旨在推进我们之前成功鉴定 sAPPα 增强子的研究。托烷司琼（F03）的目标是识别新的 sAPPα 增强剂作为新的化学实体（NCE），构成一类新型的 AD 疗法。我们坚实的初步数据基础表明该项目的具体目标很有可能实现。我们利用迭代、分层化学遗传学筛选方法从小型临床化合物库 F03 中鉴定出 F03（一种用于术后恶心和呕吐的药物），并取得了早期成功。在 AD 小鼠模型中，F03 在体外和体内均显示出持续但适度的增加，并且在口服治疗 4 周后显着改善 AD 模型的记忆力，目前 F03 正在针对轻度认知障碍 (MCI) 进行临床试验。 在这里，我们建议利用我们的高通量筛选 (HTS) 测定、迭代筛选流程和大型 UCLA 化合物库来识别 sAPPα 增强作用大于 F03 的化合物。在二次筛选中，将鉴定增加 sAPPα 的“命中”，然后使用第二个体外模型验证优先命中，然后通过离体器官型培养进一步验证。通过平行人工膜通透性分析 (PAMPA) 和体内药代动力学 (PK) 分析确定 AD 模型大脑和大脑外显率DARTS）、使用相似性整体方法进行计算机靶点识别以及新的凝胶增强靶点识别方法（GET）虽然 AD 和 MCI 的其他新治疗方法正在开发中，但仍需确定其临床结果。通过鉴定 sAPPα 增强作用大于 F03 的化合物，我们可能会发现有希望进行进一步临床开发的 NCE，也许最重要的是，我们对此类增强剂的机制研究可能会进一步阐明 sAPPα 在 AD 病因学中的作用，为 AD 病因学提供新方向。治疗的发展。