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

描述(申请人提供)：降解在阿尔茨海默病(AD)中异常堆积的淀粉样蛋白(A？)的蛋白酶，已成为体内淀粉样蛋白发生的关键调节因子，但其治疗潜力才刚刚开始探索。越来越多的实验、遗传和动物模型研究表明，胰岛素降解酶(IDE)是一种特别重要的A？降解酶。重要的是，最近的证据表明，IDE的活性可能通过几种机制中的任何一种来增加，包括内源性抑制物的置换和其分泌到细胞外空间的调节。此外，IDE的新晶体结构表明，该酶具有非常规的酶学性质，可以利用这些性质直接激活该酶40倍。在这里，我们建议使用一种针对IDE激活剂检测进行优化的基于细胞的分析方法，对包含约550,000种化合物的化学多样性文库进行超高通量筛选(UHTS)。一次化验的开发和实施将主要由佛罗里达州斯克里普斯经验丰富的uHTS Core进行，成本适中，使人们能够更多地关注对识别骨化IDE激活剂和表征其作用机制至关重要的关键二次化验(S)。我们的长期目标是确定适合在培养细胞和体内使用的药效团，这可能导致开发治疗这种毁灭性疾病的新疗法。公共卫生理由：这项提案的目标是测试一大批分子是否有可能影响已知的调控阿尔茨海默病的基本生物学过程，特别是与胰岛素降解酶有关的过程。已发现的分子将被评估其可能的治疗潜力，并可能通过未来的资金提案开发成新药。

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.