HTS for selective inhibitors of Meprin alpha and beta

 描述(申请人提供)：梅普林、和？是与系统性硬化症有关的梅津菌素蛋白水解酶超家族的成员。研究表明，meprin、和？的过度表达与皮肤和其他器官和组织中纤维状胶原沉积的增加有关。这可能会导致肾、肺和心脏纤维化，并最终导致衰竭和死亡。研究meprin、和？在系统性硬化症中的作用的主要障碍是缺乏选择性抑制剂。大多数开发的Meprin和çTate抑制剂的特点是锌结合部分以活性部位锌为靶点。大约有70种已知的人类金属蛋白水解酶的活性部位含有锌，这导致了锌结合抑制剂的非靶点毒性。根据我们的初步结果，我们假设梅普林和B选择性抑制剂将是治疗系统性硬化症的有效研究工具，其中梅普林 和？有牵连。目前还没有公开可用的Meprin的选择性抑制剂 和矿物酶。该项目的总体目标是开发梅普林、和？的选择性抑制剂。这项建议的具体目标将集中在(1)由640,000种化合物组成的SCRIPPS文库的高通量筛选；(2)Meprin、和？的选择性探针的药物化学和体外表征。我们的实验室独一无二 由于在生物学、生物化学和金属蛋白酶药物/探针发现方面的专业知识，我们有能力实现这些目标。我们发现了一类新的金属蛋白酶ADAM17抑制剂，它省去了与其最接近的类似物ADAM10和最常见的抗靶标(MMP-14和-8)。据我们所知，我们是第一个报道具有如此独特选择性的ADAM17抑制剂的实验室。此外，我们发现了另一种金属蛋白酶的新型选择性非锌结合抑制剂，即与骨关节炎有关的基质金属蛋白酶-13。与本提案最相关的是，这两项发现都是利用HTS的NIH或Tpims文库，与斯克里普斯研究所分子筛查中心(由Louis Scamvia博士和Tim Spicer先生共同领导)和Gregg Fields博士密切合作做出的。我们坚信，斯克里普斯文库的HTS将导致发现急需的Meprin、和ç的一流选择性抑制剂。我们还将与肽合成、高温超导和药物化学领域的专家合作。我们的预期结果是清楚地了解meprin和ç在系统性硬化症中的作用，这将导致对调节其进展的机制有更全面的了解，并可能导致新疗法的开发。

 DESCRIPTION (provided by applicant): Meprin  and ß are members of a superfamily of metzincin proteases implicated in systemic sclerosis. Studies indicate that the overexpression of meprin  and ß is correlated with increased deposition of fibrillar collagens in skin and other organs and tissues. This can potentially lead to kidney, lung, and heart fibrosis and eventual failure and death. The main obstacle in studying the roles of meprin  and ß in systemic sclerosis is the lack of selective inhibitors. Most of the meprin  and ß inhibitors developed t date feature Zn-binding moieties that target the active site Zn. There are approximately 70 known human metalloproteases that have Zn in their active site, which leads to an off-target toxicity of Zn-binding inhibitors. Based on our preliminary results, we hypothesize that meprin and ß selective inhibitors will be effective research tools in systemic sclerosis where meprin  and ß are implicated. There are currently no publicly available selective inhibitors of meprin  and ß metalloproteases. The overall aim of this project is to develop selective inhibitors of meprin  and ß. The specific aims of this proposal will focus on (1) High throughput screening (HTS) of the Scripps library which consists of > 640,000 compounds; (2) Medicinal chemistry and in vitro characterization of selective probes of meprin  and ß. Our laboratory is uniquely positioned to achieve these goals due to expertise in biology, biochemistry and drug/probe discovery for metalloproteases. We discovered a novel class of metalloprotease ADAM17 inhibitors that spare its closest analogue, ADAM10, and most common anti-targets (MMP-14 and -8). To our knowledge, we are the first laboratory to report ADAM17 inhibitors with such unique selectivity. Additionally, we discovered novel selective non-zinc-binding inhibitors of another metalloprotease, MMP-13, implicated in osteoarthritis. Most germane to the present proposal, both discoveries were made utilizing HTS either of NIH or TPIMS libraries in close collaboration with Scripps Research Institute Molecular Screening Center (co-headed by Dr. Louis Scampavia and Mr. Tim Spicer) and Dr. Gregg Fields. We strongly believe that HTS of Scripps library will result in the discovery of much needed first-in-class selective inhibitors of meprin  and ß. We will also collaborate with experts in the fields of peptide synthesis, HTS, and medicinal chemistry. Our expected outcome is a clear understanding of the role of meprin  and ß in systemic sclerosis, which will lead to a more comprehensive knowledge of the mechanisms that regulate its progression and potentially result in the development of novel therapies.