HTS for Modulators of Beta-Amyloid Catabolism by Insulin-Degrading Enzyme

HTS 用于调节胰岛素降解酶 β-淀粉样蛋白分解代谢

• 批准号: 7559775
• 负责人: MALCOLM A LEISSRING
• 金额: $ 2.5万
• 依托单位: MAYO CLINIC JACKSONVILLE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7559775
• 关键词: Address; Alkylation; Alzheimer' s Disease; Amyloid; Amyloid Proteins; Amyloid beta-Protein Precursor; Animal Model; Appendix; Behavioral; Bioavailable; Biological Assay; Biology; Brain; Brain region; Catabolism; Cell Line; Cells; Cerebrum; Chemicals; Clinical Trials; Cognition; Collection; Cultured Cells; Defect; Development; Disease; Endopeptidases; Equilibrium; Evaluation; Extracellular Space; Florida; Fluorescence; Fluorescence Polarization; Genetic; Goals; Health; Hela Cells; Human; Insulinase; Knock-out; Laboratories; Memory; Metalloproteases; Monoclonal Antibodies; Mouse Cell Line; Mutation; Pathogenesis; Pathology; Pathway interactions; Peptide Hydrolases; Personal Satisfaction; Physiological; Production; Property; Proteins; Purpose; Range; Rate; Reagent; Relative (related person); Research; Role; Screening procedure; Series; Site; Specificity; Structure; Sulfhydryl Compounds; Time; Transgenic Organisms; Up-Regulation; Work; Zinc; amyloidogenesis; base; chemical addition; enzyme activity; extracellular; high throughput screening; in vivo; inhibitor/antagonist; man; member; novel; pharmacophore; pre-clinical; prevent; repository; research study; secretase; small molecule; small molecule libraries; therapeutic target; tool; trafficking

DESCRIPTION (provided by applicant): Alzheimer's disease (AD) is characterized by abnormal accumulation of the amyloid ?-protein (A?) in brain regions subserving memory and cognition. In recent years, proteases that degrade A??have been identified as potent and rate-limiting regulators of cerebral A??levels and amyloidogenesis in vivo. Significantly, orally bioavailable compounds that activate A?-degrading proteases have already been identified that are effective in reducing AD-type pathology in animal models and are currently entering clinical trials. Converging lines of evidence strongly implicate insulin-degrading enzyme (IDE) as a particularly important A?-degrading protease. Nonetheless, there is a surprising lack of pharmacological tools targeting IDE, or indeed any member of the unusual zinc-metalloprotease superfamily to which it belongs. Importantly, accumulating evidence shows how IDE activity might be augmented pharmacologically by any of several mechanisms, including the displacement of endogenous inhibitors or modulation of its expression and/or secretion into the extracellular space. Moreover, new crystal structures of IDE show that this protease possesses unorthodox structural features that can be targeted to directly activate the protease as much as 4000 percent. The purpose of this proposal is to utilize our well-characterized fluorescence polarization-based A?-degradation assay (Leissring et al., JBC 2003, Appendix) to search for chemical modulators of IDE within the Small Molecule Library of the MLSCN. We propose a series of secondary assays to confirm discovered hits, to establish their specificity for IDE, and to identify cell-penetrant compounds. Probes suitable for use in cultured cells or in vivo will be used in downstream experiments to resolve several outstanding questions about the role of IDE in AD pathogenesis that can only be addressed via a chemical biology approach. In addition, chemical activators of IDE with suitable properties could serve as pharmacophores for the development of novel AD therapies.

描述（由申请人提供）：阿尔茨海默病（AD）的特征是淀粉样蛋白？蛋白质（A？）大脑中负责记忆和认知的区域。近年来，降解A？已被确定为有效的和限速调节大脑A？？水平和体内淀粉样蛋白生成。值得注意的是，口服生物可利用的化合物，激活A？-已经鉴定了降解蛋白酶，其在动物模型中有效减少AD型病理，并且目前正在进入临床试验。越来越多的证据表明，胰岛素降解酶（IDE）是一种特别重要的A？降解蛋白酶。尽管如此，令人惊讶的是缺乏针对IDE的药理学工具，或者确实缺乏它所属的不寻常的锌金属蛋白酶超家族的任何成员。重要的是，越来越多的证据表明IDE活性可能通过几种机制中的任何一种来增强，包括内源性抑制剂的置换或其表达和/或分泌到细胞外空间的调节。此外，IDE的新晶体结构表明，这种蛋白酶具有非正统的结构特征，可以靶向直接激活蛋白酶高达4000%。本提案的目的是利用我们表征良好的基于荧光偏振的A？-降解测定（Leissring等人，JBC 2003，附录），以在MLSCN的小分子文库中搜索IDE的化学调节剂。我们提出了一系列的二次检测，以确认发现的命中，建立其特异性IDE，并确定细胞渗透剂化合物。适用于培养细胞或体内的探针将用于下游实验，以解决有关IDE在AD发病机制中的作用的几个悬而未决的问题，这些问题只能通过化学生物学方法解决。此外，具有合适性质的IDE的化学活化剂可作为开发新型AD疗法的药效团。

项目成果

The blood glucose-lowering effect of racecadotril is not attributable to inhibition of insulin-degrading enzyme.

消旋卡多曲的降血糖作用并非归因于对胰岛素降解酶的抑制。

• DOI: 10.1055/s-0033-1353211
• 发表时间: 2014
• 期刊: Hormone and metabolic research = Hormon- und Stoffwechselforschung = Hormones et metabolisme
• 作者: Kurklinsky,S;Abdul-Hay,SO;McGuire,MP;Howard,EA;Knight,J;Leissring,MA
• 通讯作者: Leissring,MA