High-throughput compound screening for modulators of insulin-degrading enzyme.

胰岛素降解酶调节剂的高通量化合物筛选。

基本信息

批准号：
7619035
负责人：
MALCOLM A LEISSRING
金额：
$ 18.82万
依托单位：
MAYO CLINIC JACKSONVILLE
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-05-15 至 2011-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7619035
关键词：
AD 20 Affect Alzheimer&apos s Disease Amyloid Amyloid Proteins Amyloid beta-Protein Precursor Animal Model Attention Behavioral Biological Assay Biological Process Catabolism Cells Collection Cultured Cells Defect Detection Development Disease Enzyme Activators Equilibrium Experimental Genetics Extracellular Space Florida Fluorescence Polarization Funding Future Genetic Goals Hela Cells Housing Human In Vitro Insulinase Laboratories Lead Libraries Mediating Miniaturization Mutation Nature Neurons Pathology Peptide Hydrolases Permeability Pharmaceutical Chemistry Pharmaceutical Preparations Plasmin Plasminogen Activator Inhibitor 1 Post-Translational Protein Processing Preparation Process Production Property Proteins Public Health Recombinants Resources Robotics Screening procedure Structure Substrate Specificity System Testing Therapeutic Transgenic Organisms Work amyloidogenesis assay development base bone chemical genetics cost counterscreen cytotoxicity design drug discovery enzyme activity experience high throughput screening in vivo inhibitor/antagonist man novel overexpression pharmacophore prevent secretase small molecule therapeutic target trafficking

项目摘要

DESCRIPTION (provided by applicant): Proteases that degrade the amyloid ¿-protein (A¿), which accumulates abnormally in Alzheimer's disease (AD), have emerged as critical regulators of amyloidogenesis in vivo, yet have only begun to be explored for their therapeutic potential. Accumulating experimental, genetic and animal modeling studies implicate insulin-degrading enzyme (IDE), as a particularly important A¿-degrading protease. Importantly, recent evidence shows how IDE activity might be increased by any of several mechanisms, including the displacement of endogenous inhibitors and modulation of its secretion into the extracellular space. Moreover, new crystal structures of IDE show that this protease possesses unorthodox enzymological properties that can be exploited to directly activate the protease as much as 40-fold. Here we propose to conduct ultra high- throughput screening (uHTS) on a chemically diverse library of ~550,000 compounds using a cell-based assay optimized for the detection of IDE activators. The development and implementation of the primary assay will largely be performed by Scripps Florida's highly experienced uHTS Core for a modest cost, permitting greater attention to be focused on critical secondary assays essential for identifying bone fide IDE activators and characterizing their mechanism(s) of action. Our long-term goal is to identify pharmacophores suitable for use in cultured cells and in vivo, which may lead to the development of novel therapies to treat this devastating disease. PUBLIC HEALTH RELELVANCE: The goal of this proposal is to test a large collection of molecules for their potential to affect fundamental biological processes known to regulate Alzheimer's disease, specifically relating to insulin-degrading enzyme. Discovered molecules will be evaluated for their possible therapeutic potential, and potentially developed into novel drugs through future funding proposals.

描述（应用程序提供）：降解淀粉样蛋白» - 蛋白质（A e）的蛋白酶，它在阿尔茨海默氏病中累积了绝对的蛋白质（AD），它已成为体内淀粉样生成的关键调节剂，但才开始探索其治疗潜力。累积实验性，遗传和动物建模研究暗示胰岛素降解酶（IDE）是一种特别重要的蛋白酶。重要的是，最近的证据表明，如何通过几种机制中的任何一种来增加IDE活性，包括内源性抑制剂的位移以及对细胞外空间的分泌调节。此外，IDE的新晶体结构表明，该蛋白酶具有非正统的酶学特性，可以探索以直接激活蛋白酶40倍。在这里，我们建议使用优化用于检测IDE激活剂的基于细胞的基于细胞的测定法，对〜550,000种化合物的化学多样性库进行超高通量筛选（UHT）。主要测定法的开发和实施将在很大程度上由佛罗里达州经验丰富的UHTS核心的核心进行适度的成本进行，从而更加关注关键的辅助测定法，这对于识别骨骼的fide IDE激活剂和表征其作用机制所必需的至关重要。我们的长期目标是鉴定适合在培养细胞和体内使用的药体，这可能导致开发用于治疗这种毁灭性疾病的新型疗法。公共卫生相关：该提案的目的是测试大量分子，以影响已知的调节阿尔茨海默氏病的基本生物学过程的潜力，特别与胰岛素降解酶有关。发现的分子将根据其可能的治疗潜力进行评估，并有可能通过未来的资助建议将其发展为新的药物。