Structure, Function and Application of Metalloproteinase Inhibitors in Osteoarthr

金属蛋白酶抑制剂的结构、功能及其在骨关节炎中的应用

基本信息

批准号：
8457990
负责人：
KEITH BREW
金额：
$ 45.55万
依托单位：
FLORIDA ATLANTIC UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1991
资助国家：
美国
起止时间：
1991-09-01 至 2016-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8457990
关键词：
Affect Age Animal Disease Models Animal Model Antibodies Binding Biochemical Calcium Cartilage Cartilage Matrix Clinical Collaborations Collagen Complex Crotalus adamanteus proteinase II Degenerative polyarthritis Disease Economic Burden Elderly Engineering Enzymes Extracellular Matrix Funding Germany Goals Health Human In Vitro Institutes Interstitial Collagenase Intervention Joints Libraries Ligaments Matrix Metalloproteinases Medial Metalloproteases Methods Molecular Mus Mutation Operative Surgical Procedures Pentosan Polysulfate Peptide Hydrolases Phage Display Play Population Protein Engineering Proteins Proteomics Replacement Arthroplasty Research Societies Specificity Stromelysin 1 Structure System TNF-alpha converting enzyme Testing Therapeutic Agents Therapeutic Intervention Tissue Engineering Tissue Inhibitor of Metalloproteinase-1 Tissue Inhibitor of Metalloproteinase-3 Tissue Inhibitor of Metalloproteinases Transgenic Mice Tumor Necrosis Factor-alpha Use Effectiveness Variant aggrecan aggrecanase arthropathies articular cartilage base collagenase collagenase 3 design economic cost effective therapy efficacy testing human tissue in vivo in vivo Model inhibitor/antagonist mouse model mutant novel novel strategies novel therapeutics overexpression prevent prototype screening socioeconomics therapeutic development therapeutic target

项目摘要

DESCRIPTION (provided by applicant): Osteoarthritis (OA) is the most prevalent joint disease affecting most of the elderly, imposing a major socio- economic burden on society. The hallmark of the disease is the breakdown of articular cartilage by elevated proteinase activities that degrade major extracellular matrix molecules, aggrecans and collagens, but currently there are no effective treatments for OA, except joint replacement surgery. Our long-term objective is to find ways to specifically inhibit these proteinases and apply them for clinical intervention in OA. The key enzymes involved are matrix metalloproteinases (MMPs) and adamalysins with thrombospondin motif (ADAMTSs). Their activities are regulated by the endogenous tissue inhibitors of metalloproteinases (TIMPs). Our recent studies have shown that TIMP-3 variants that selectively inhibit aggrecanases prevent the progression of OA in an animal model, whereas wild-type TIMP-3 that broadly inhibits MMPs and ADAMTSs was not effective. This suggests that highly selective inhibitors are essential for therapeutic development. To achieve this goal two major aims are proposed. The first is to understand the molecular basis for selectivity in our TIMP-3 variants using biochemical, biophysical and structural methods, and to use this to further develop highly discriminating inhibitors of metalloproteinases. Specifically, we will rationally design TIMP-3 variants that differentially inhibit the main two aggrecanases, ADAMTS-4 and -5, and screen human antibody phage display libraries for "exosite inhibitors" of aggrecanases and collagenases. The prototype exosite inhibitors will be further refined by mutation. The mechanism by which calcium pentosan polysulfate (CaPPS), an exosite inhibitor of aggrecanases, enhances the interactions between TIMP-3 and aggrecanases will be also elucidated. The second major aim is to evaluate the efficacy of available and newly developed metalloproteinase inhibitors using in vitro and in vivo models of OA and to validate target enzymes in human OA using these inhibitors. To this end, we will continue characterizing TIMP-3 and [-1A]TIMP-3 transgenic mice to delineate the molecular basis of their differential effects on blocking OA progression by analyzing extracellular matrix components protected by these inhibitors using a combination of immunological and proteomic approaches. Newly developed exosite inhibitors and TIMP variants will be tested for their efficacy to protect cartilage from degradation using an in vitro cartilage explant culture system and the in vivo mouse model of OA induced by medial meniscotibial ligament transaction. Effective inhibitors will be also tested in human cartilage in culture to further validate their efficacy. Using available inhibitors we will identify and characterize a novel TIMP-1- sensitive aggrecanase as it is considered as a potential therapeutic target in humans. Accomplishment of these goals will provide new principles for developing therapeutic interventions for OA.

描述（由申请人提供）：骨关节炎（OA）是影响大多数老年人的最常见的关节疾病，给社会带来重大的社会经济负担。该疾病的特点是蛋白酶活性升高，从而降解主要的细胞外基质分子、聚集蛋白聚糖和胶原蛋白，从而破坏关节软骨，但目前除了关节置换手术外，还没有有效的治疗 OA 的方法。我们的长期目标是找到特异性抑制这些蛋白酶的方法，并将其应用于骨关节炎的临床干预。涉及的关键酶是基质金属蛋白酶 (MMP) 和具有血小板反应蛋白基序的金刚烷胺 (ADAMTS)。它们的活性受到内源性金属蛋白酶组织抑制剂 (TIMP) 的调节。我们最近的研究表明，选择性抑制软骨聚集蛋白聚糖酶的 TIMP-3 变体可预防动物模型中 OA 的进展，而广泛抑制 MMP 和 ADAMTS 的野生型 TIMP-3 则无效。这表明高选择性抑制剂对于治疗开发至关重要。为了实现这一目标，提出了两个主要目标。首先是使用生物化学、生物物理和结构方法了解我们的 TIMP-3 变体选择性的分子基础，并利用它进一步开发高度区分的金属蛋白酶抑制剂。具体来说，我们将合理设计能够差异性抑制两种主要聚集蛋白聚糖酶ADAMTS-4和-5的TIMP-3变体，并筛选人抗体噬菌体展示文库以寻找聚集蛋白聚糖酶和胶原酶的“外部抑制剂”。原型外位点抑制剂将通过突变进一步完善。戊聚糖多硫酸钙 (CaPPS)（一种聚集蛋白聚糖酶的外部位点抑制剂）增强 TIMP-3 和聚集蛋白聚糖酶之间相互作用的机制也将得到阐明。第二个主要目标是使用 OA 的体外和体内模型评估现有和新开发的金属蛋白酶抑制剂的功效，并使用这些抑制剂验证人类 OA 中的靶酶。为此，我们将继续表征 TIMP-3 和 [-1A]TIMP-3 转基因小鼠，通过结合免疫学和蛋白质组学分析受这些抑制剂保护的细胞外基质成分，描绘它们对阻止 OA 进展的不同作用的分子基础。接近。新开发的外位点抑制剂和 TIMP 变体将使用体外软骨外植体培养系统和内侧半月板韧带交易诱导的体内 OA 小鼠模型来测试其保护软骨免遭降解的功效。有效的抑制剂还将在人类软骨培养物中进行测试，以进一步验证其功效。使用现有的抑制剂，我们将鉴定并表征一种新型 TIMP-1 敏感的聚集蛋白聚糖酶，因为它被认为是人类的潜在治疗靶点。这些目标的实现将为开发 OA 治疗干预措施提供新原则。

项目成果

期刊论文数量（87）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Generation of a Mouse Line Harboring a Bi-Transgene Expressing Luciferase and Tamoxifen-Activatable CreERT2 Recombinase in Cartilage

DOI：
10.1002/dvg.22734
发表时间：
2014-02-01
期刊：
GENESIS
影响因子：
1.5
作者：
Lo Cascio, Leandro;Liu, Ke;Bou-Gharios, George
通讯作者：
Bou-Gharios, George

The C-terminal domains of ADAMTS-4 and ADAMTS-5 promote association with N-TIMP-3.

DOI：
10.1016/j.matbio.2009.07.005
发表时间：
2009-10
期刊：
MATRIX BIOLOGY
影响因子：
6.9
作者：
Troeberg, Linda;Fushimi, Kazunari;Scilabra, Simone D.;Nakamura, Hiroyuki;Dive, Vincent;Thogersen, Ida B.;Enghild, Jan J.;Nagase, Hideaki
通讯作者：
Nagase, Hideaki

Development of a monoclonal anti-ADAMTS-5 antibody that specifically blocks the interaction with LRP1.

开发特异性阻断与 LRP1 相互作用的单克隆抗 ADAMTS-5 抗体。

DOI：
10.1080/19420862.2017.1304341
发表时间：
2017
期刊：
mAbs
影响因子：
5.3
作者：
Santamaria,Salvatore;Fedorov,Oleg;McCafferty,John;Murphy,Gillian;Dudhia,Jayesh;Nagase,Hideaki;Yamamoto,Kazuhiro
通讯作者：
Yamamoto,Kazuhiro

Aggrecanases and cartilage matrix degradation.