New probes for matrix metalloproteinase 13

基质金属蛋白酶 13 的新探针

• 批准号: 8631897
• 负责人: GREGG B FIELDS
• 金额: $ 50.31万
• 依托单位: TORREY PINES INST FOR MOLECULAR STUDIES
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-17 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8631897
• 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

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）是关节炎的最常见形式，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Aggrecan，硫酸软骨素和透明质酸。基质金属蛋白酶13（MMP-13）已被证明是主要胶原酶，导致OA期间关节软骨降解。多次尝试开发基于MMP-13抑制剂的药物的尝试主要是由于剂量限制副作用统称为肌肉骨骼综合征（MSS）。虽然尚不清楚MSS的确切原因是由于对金属蛋白酶家族的其他代表缺乏候选药物的选择性。鉴定蛋白酶二级结合位点（外粒子），即促进或调节蛋白酶活性的非活性部位区域，可以用于设计蛋白酶家族中选择性抑制剂。我们的实验室已经开发了三螺旋肽（THP）荧光共振能量转移（FRET）底物，用于胶原式MMP的高吞吐量筛选（HTS），并鉴定了推定的外部结合化合物。使用带有FRET THP测定的HTS技术，我们先前鉴定出MMP-13的几种选择性低微摩尔抑制剂。一种抑制剂（化合物Q/4，Pubchem CID 2047223）表现出特性，表明其作用方式不是通过Zn螯合。化合物Q/4的分子支架比以前描述的MMP-13抑制剂的分子支架要小得多，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，如体内MMP-13探针。实现此目标的具体目的如下：（1）X射线晶体学引导的MMP-13外部探针的药物化学； （2）MMP-13外部探针的体外和体内测试。目前的工作将导致体内探针，可以评估OA中选择性抑制剂对细胞外蛋白水解活性的影响，并为开发新型MMP成像剂的开发奠定了基础。另外，将确定一种独特的MMP抑制模式。