Development of small molecule probes of matrix metallo protease function

基质金属蛋白酶功能小分子探针的研制

• 批准号: 7979219
• 负责人: Matthew Bogyo
• 金额: $ 24万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7979219
• 关键词: Active Sites; Amino Acids; Binding; Biology; Catalytic Domain; Cells; Clinical Trials; Complex; Cysteine; Data; DegP protease; Development; Disease; Engineering; Enzymes; Family; Family member; Financial compensation; Generations; Image; Individual; Knock-out; Label; Link; Malignant Neoplasms; Marketing; Metals; Methodology; Methods; Modification; Molecular; Molecular Models; Monitor; Mutation; Nature; Pathogenesis; Pathology; Peptide Hydrolases; Peptides; Pharmaceutical Chemistry; Pharmaceutical Preparations; Rage; Reporter; Role; Side; System; Time; Zinc; base; chemotherapy; complex biological systems; design; high risk; human disease; hydroxamate; imaging probe; in vivo; inhibitor/antagonist; member; molecular modeling; mutant; novel strategies; public health relevance; scaffold; small molecule; tumor growth

DESCRIPTION (provided by applicant): The matrix metallo proteases (MMPs) are a large family of enzymes that have been found to regulate the pathogenesis of a wide rage of human diseases, most notably cancer. Their potential role in matrix degradation during tumor growth originally made them promising targets for the development of new chemotherapy drugs. However, an overall lack of understanding of the roles of specific members of this protease family in cancer pathology resulted in a large number of failed clinical trials and no new MMP drugs entering the market. One of the confounding issues that limited efforts to better define MMP function is the overall large size of the metallo protease family and a lack of highly selective inhibitors for temporally controlled inhibition of specific MMP family members. Furthermore, efforts to genetically knock-out specific proteases have been limited by the essential nature of many of these proteases and the potential for compensation by related family members. Thus, high risk and non-traditional methods to both selectively inhibit and image to dynamics of localization of individual MMP proteases will be required to begin to understand specific functional roles of each MMP family member in the pathogenesis of human diseases such as cancer. This proposal outlines our plans to develop a novel strategy to develop small molecules that can be used to both selectively inhibit and also dynamically image individual MMP proteases in the context of a complex biological system. This method is based on the engineering of specific MMP targets to contain a reactive cysteine residue near the active site that can be used for direct covalent modification by a probe containing a reactive electrophile. Inhibitors that bind to the metal in the active site and have the ability to covalently link to the engineered cysteine residue can then be used to selectively target the Cys-mutant MMPs. This allows selective covalent inhibition and labeling of a single MMP target. At the completion of this project we plan to have the method validated for multiple MMP targets such that it can then be applied to longer term projects designed to further dissect MMP biology. PUBLIC HEALTH RELEVANCE: This project outlines plans to develop a general method to engineer matrix metallo proteases (MMPs) such that they can be selectively inhibited and imaged using small molecule activity based probes. This methodology, once applied to complex in vivo system, will allow studies of the involvement of individual MMP proteases in disease pathology.

描述（由申请人提供）：基质金属蛋白酶（MMP）是一个大家族的酶，已发现其调节多种人类疾病（最显著的是癌症）的发病机理。它们在肿瘤生长过程中基质降解中的潜在作用最初使它们成为开发新化疗药物的有希望的靶点。然而，对该蛋白酶家族的特定成员在癌症病理学中的作用总体缺乏了解，导致大量临床试验失败，并且没有新的MMP药物进入市场。限制更好地定义MMP功能的努力的混杂问题之一是金属蛋白酶家族的整体大尺寸和缺乏用于暂时控制抑制特定MMP家族成员的高度选择性抑制剂。此外，基因敲除特定蛋白酶的努力受到许多这些蛋白酶的基本性质和相关家族成员补偿的可能性的限制。因此，需要选择性抑制和成像于单个MMP蛋白酶的定位动力学的高风险和非传统方法，以开始理解每个MMP家族成员在人类疾病如癌症的发病机制中的特定功能作用。该提案概述了我们的计划，以开发一种新的策略，开发小分子，可用于选择性抑制和动态成像的背景下，一个复杂的生物系统中的个别MMP蛋白酶。该方法基于特定MMP靶标的工程改造，以在活性位点附近含有反应性半胱氨酸残基，其可用于通过含有反应性亲电试剂的探针进行直接共价修饰。然后可以使用与活性位点中的金属结合并且具有与工程化半胱氨酸残基共价连接的能力的抑制剂来选择性地靶向Cys突变MMP。这允许选择性共价抑制和标记单个MMP靶标。在这个项目完成后，我们计划有多个MMP目标的方法验证，使它可以应用于长期项目，旨在进一步剖析MMP生物学。 公共卫生相关性：该项目概述了计划开发一种通用方法来工程化基质金属蛋白酶（MMP），以便使用基于小分子活性的探针选择性抑制和成像。这种方法，一旦应用于复杂的体内系统，将允许研究的参与个别MMP蛋白酶在疾病的病理。