Screening for enhancers of sAPPalpha

筛选 sAPPalpha 增强子

基本信息

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
项目类别：
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&apos s Disease Alzheimer&apos 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

项目摘要

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（由适用提供）：阿尔茨海默氏病（AD）目前以每年超过2000亿美元的价格折磨于美国540万人。目前批准的药物仅提供短期症状缓解，但不会改变疾病进展。是淀粉样蛋白蛋白（APP）的淀粉样蛋白β胡椒（Aβ）的产生增加和/或清除率增加的AD脑标志的神经斑。应用可以通过两种途径处理。在一个中，它经历了顺序的β-分泌酶和γ-分泌酶的切割，以产生固定淀粉样蛋白前体蛋白β（SAPPβ）和Aβ；在另一种中，它被α-分泌酶裂解以产生固体淀粉样蛋白前体蛋白α（SAPPα），这是一种可能阻止AD的促肽。该项目的提议旨在推进我们以前的研究，该研究成功地识别了SAPPα-增强剂Tropisetron（F03），目的是将新的SAPPα-增强剂识别为新的化学实体（NCES），构成了一种新型的AD治疗疗法。我们的初步数据的强大基础表明，可以实现该项目的具体目的很高。我们在识别F03（一种用于手术后恶心和呕吐的药物）中，利用我们的迭代，分层化学基因筛查方法获得了早期的成功。在AD的小鼠模型中，F03在体外和体内持续但适度地增加了SAPPα，并在口服四个星期后显着改善AD模型的记忆。 F03现在正在接受轻度认知障碍的临床试验（MCI）作为重新利用的药物进行广告。在这里，我们建议利用我们的高通量筛选（HTS）测定，迭代筛选流量和大型UCLA化合物库来识别具有SAPPα增强效果的化合物大于F03。在主要的HTS屏幕中，将确定增加SAPPα的“命中”。在次要屏幕中，使用第二个体外模型验证了命中。然后，通过平行人工膜渗透率分析（PAMPA）和体内药代动力学（PK）分析确定的AD模型脑和大脑渗透率的离体有机培养将进一步验证优先级的命中。最后，我们将使用CEREP BIOPRINT曲线，药物亲和力响应靶标稳定性（DARTS），使用相似性集合方法以及通过新的凝胶增强靶标识别方法（GET）在硅靶ID中诱导SAPPα增强的机制。尽管正在开发其他新方法来识别AD和MCI治疗疗法，但它们的临床结果仍然不确定。通过鉴定具有SAPPα增强作用的化合物大于F03，我们可能会发现有希望的进一步临床发育。也许最重要的是，我们对此类增强剂的机械研究可能会进一步阐明SAPPα在AD病因中的作用，从而为治疗性发育提供了新的方向。