New probes for matrix metalloproteinase 13

基质金属蛋白酶 13 的新探针

基本信息

批准号：
8737007
负责人：
GREGG B FIELDS
金额：
$ 40.54万
依托单位：
TORREY PINES INST FOR MOLECULAR STUDIES
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-17 至 2015-01-05
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8737007
关键词：
Active Sites Adverse effects Affinity Arthritis Back Binding Binding Sites Biological Assay Cartilage Chondroitin Sulfates Collagen Collagen Type II Compound Q Cytochrome P450 Degenerative polyarthritis Development Dose-Limiting Enzymes Exhibits Family Family member Fluorescence Resonance Energy Transfer Goals Hyaluronan Image In Vitro Joints Laboratories Lead Matrix Metalloproteinases Metalloproteases Modeling Modification Molecular Molecular Models Musculoskeletal Neutrophil Collagenase New Agents Peptide Hydrolases Peptides Pharmaceutical Chemistry Pharmaceutical Preparations Property Proteoglycan PubChem Rattus Reporting Research Proposals Roentgen Rays Site Specificity Staging Structure Syndrome Techniques Tensile Strength Testing Therapeutic Toxic effect United States National Institutes of Health Work X-Ray Crystallography aggrecan analog articular cartilage base chelation collagenase collagenase 3 design drug candidate extracellular feeding high throughput screening improved in vivo inhibitor/antagonist molecular modeling novel prevent programs public health relevance scaffold success

项目摘要

DESCRIPTION (provided by applicant): Osteoarthritis (OA), the most common form of arthritis, is characterized by the destruction of articular cartilage. The main constituents of articular or joint cartilage are type II collagen and various proteoglycans, such as aggrecan, chondroitin sulfate, and hyaluronan. Matrix metalloproteinase 13 (MMP-13) has been shown to be main collagenase responsible for degradation of articular cartilage during OA. Multiple attempts to develop an MMP-13 inhibitor-based drug failed mostly due to the dose limiting side effects collectively known as musculoskeletal syndrome (MSS). While the exact cause of MSS is not known it is believed to be due to the lack of selectivity of drug candidates towards other representatives of metalloproteinase families. Identification of protease secondary binding sites (exosites), i.e. non-active site regions that facilitate or modulate protease activity, could be utilized for the design of selective inhibitors within protease families. Our laboratory has developed triple-helical peptide (THP) fluorescence resonance energy transfer (FRET) substrates for high throughput screening (HTS) of collagenolytic MMPs and identification of putative exosite-binding compounds. Using HTS techniques with FRET THP assays, we previously identified several selective low micromolar inhibitors of MMP-13. One inhibitor (compound Q/4, PubChem CID 2047223) exhibited properties suggesting that its mode of action was not via Zn chelation. Compound Q/4 possessed a significantly smaller molecular scaffold than previously described MMP-13 inhibitors, yet offered excellent selectivity. We wish to further develop compound Q/4 and its analogs as in vivo MMP-13 probes. The specific aims to achieve this goal are as follows: (1) X-ray crystallography-guided medicinal chemistry of MMP-13 exosite probes; and (2) in vitro and in vivo testing of MMP-13 exosite probes. The present work will lead to in vivo probes that can evaluate the effects of selective inhibitors on extracellular proteolytic activity in OA and lay the groundwork for the development of novel MMP imaging agents. In addition, a unique mode of MMP inhibition will be identified.

描述（由申请人提供）：骨关节炎（OA）是最常见的关节炎形式，其特征是关节软骨的破坏。关节或关节软骨的主要成分是II型胶原蛋白和各种蛋白聚糖，例如聚集蛋白聚糖、硫酸软骨素和透明质酸。基质金属蛋白酶 13 (MMP-13) 已被证明是导致 OA 期间关节软骨降解的主要胶原酶。开发基于 MMP-13 抑制剂的药物的多次尝试都失败了，主要是由于剂量限制性副作用，统称为肌肉骨骼综合征 (MSS)。虽然 MSS 的确切原因尚不清楚，但据信是由于候选药物对金属蛋白酶家族的其他代表缺乏选择性。蛋白酶二级结合位点（外位点）的鉴定，即促进或调节蛋白酶活性的非活性位点区域，可用于设计蛋白酶家族内的选择性抑制剂。我们的实验室开发了三螺旋肽 (THP) 荧光共振能量转移 (FRET) 底物，用于溶胶原 MMP 的高通量筛选 (HTS) 和鉴定假定的外部位点结合化合物。使用 HTS 技术和 FRET THP 测定，我们之前鉴定了几种选择性低微摩尔 MMP-13 抑制剂。一种抑制剂（化合物 Q/4，PubChem CID 2047223）表现出的特性表明其作用方式不是通过 Zn 螯合。化合物 Q/4 具有比之前描述的 MMP-13 抑制剂显着更小的分子支架，但具有出色的选择性。我们希望进一步开发化合物Q/4及其类似物作为体内MMP-13探针。实现这一目标的具体目标如下：（1）X射线晶体学引导MMP-13外位点探针的药物化学； (2) MMP-13 外部位点探针的体外和体内测试。目前的工作将导致体内探针能够评估选择性抑制剂对 OA 细胞外蛋白水解活性的影响，并为新型 MMP 显像剂的开发奠定基础。此外，还将确定一种独特的 MMP 抑制模式。