Structure-function studies on human Angiotensin-I converting enzyme (ACE)

血管紧张素- 1转换酶[ACE，含有两个结构域（N和C）]抑制剂被广泛用于治疗心血管疾病，包括高血压、心力衰竭、冠状动脉疾病、纤维化和肾衰竭。然而，当前一代的ACE抑制剂是在20世纪70年代和80年代开发的，由于常见的副作用而受到阻碍。ACE的N-和c -结构域表现出不同的底物特异性。虽然市场上有许多ACE抑制剂可以阻断这两个结构域，但目前还没有一种药物可以选择性地抑制n结构域，从而获得减少心脏、肾脏和肺部纤维化和炎症的优势，而不会伴随阻断c结构域引起的副作用。这强调了确定ACE三维结构和设计更安全、更有效的第二代ACE抑制剂复合物的重要性。我们成功测定了人睾丸ACE的晶体结构（相当于体细胞ACE的C结构域）和体细胞ACE的N结构域（Angiotensin-I converting enzyme [ACE，其中包含两个结构域（N和C））)抑制剂，广泛用于治疗心血管疾病，包括高血压、心力衰竭、冠状动脉疾病、纤维化和肾衰竭。然而，当前一代的ACE抑制剂是在20世纪70年代和80年代开发的，由于常见的副作用而受到阻碍。ACE的N-和c -结构域表现出不同的底物特异性。虽然市场上有许多ACE抑制剂可以阻断这两个结构域，但目前还没有一种药物可以选择性地抑制n结构域，从而获得减少心脏、肾脏和肺部纤维化和炎症的优势，而不会伴随阻断c结构域引起的副作用。这强调了确定ACE三维结构和设计更安全、更有效的第二代ACE抑制剂复合物的重要性。我们利用x射线晶体学成功测定了人类睾丸ACE的晶体结构（相当于体细胞ACE的C结构域）和体细胞ACE的n结构域（具有各种临床重要抑制剂以及新型结构域选择性抑制剂），为真正基于结构设计更好的ACE抑制剂提供了平台。这是在蛋白酶的结构生物学方面的重大突破，更重要的是，ACE抑制机制的突破。这为一种更严格的方法铺平了道路，通过基于结构的新型结构域选择性抑制剂的药物设计方法来利用结构域之间的差异。因此，在ACE方面的持续努力，为我们组开展新的研究提供了坚实的平台。我们提出的实验建立在先前和当前工作的基础上，旨在研究全长体ACE和ACE与区域选择性抑制剂复合物的晶体结构，并结合对一个重要医学问题的基础和转化研究。从长远来看，该申请旨在利用详细的结构知识来合成新的ACE抑制剂，目的是为治疗心血管疾病，特别是高血压和纤维化提供更好的药物。一个关键特征将是设计针对ACE的N-或c端结构域的特异性化合物，期望这将提供具有较少副作用的选择性治疗化合物。

Angiotensin-I converting enzyme [ACE, which contains two domains (N and C)] inhibitors are widely used to treat cardiovascular diseases, including high blood pressure, heart failure, coronary artery disease, fibrosis and kidney failure. However, current-generation ACE inhibitors, which were developed in the 1970's and 1980's, are hampered by common side effects. The N- and C-domains of ACE display different substrate specificities. While there are many ACE inhibitors on the market that block both domains, there are no drugs that selectively inhibit the N-domain and thereby accrue the advantages of reducing fibrosis and inflammation in the heart, kidney and lung, without the concomitant side effects induced by blockade of the C-domain. This underscores the importance of the determination of the 3D structure of ACE and the design of 2nd generation ACE-inhibitor complex/s that are safer and more effective. Our success in the determination of the crystal structure of human testis ACE (equivalent to the C-domain of somatic ACE) and the N-domain of somatic ACE (Angiotensin-I converting enzyme [ACE, which contains two domains (N and C)] inhibitors are widely used to treat cardiovascular diseases, including high blood pressure, heart failure, coronary artery disease, fibrosis and kidney failure. However, current-generation ACE inhibitors, which were developed in the 1970's and 1980's, are hampered by common side effects. The N- and C-domains of ACE display different substrate specificities. While there are many ACE inhibitors on the market that block both domains, there are no drugs that selectively inhibit the N-domain and thereby accrue the advantages of reducing fibrosis and inflammation in the heart, kidney and lung, without the concomitant side effects induced by blockade of the C-domain. This underscores the importance of the determination of the 3D structure of ACE and the design of 2nd generation ACE-inhibitor complex/s that are safer and more effective. Our success in the determination of the crystal structure of human testis ACE (equivalent to the C domain of somatic ACE) and the N-domain of somatic ACE (with various clinically important inhibitors as well as novel domain selective inhibitors) using X-ray crystallography have provided the platform for true structure-based design of better ACE inhibitors. This is a significant breakthrough in terms of the structural biology of the protease and, more importantly, the mechanism of ACE inhibition. This paves the way for a more rigorous approach exploiting the differences between the domains through a structure based drug design approach of novel domain-selective inhibitors. Thus the sustained effort on ACE has provided a firm platform for our group for the new studies. Our proposed experiments that builds on a body of previous and current work are directed at structural study of the full-length somatic ACE and crystal structures of complexes of ACE with domain selective inhibitors combining basic and translational research on an important medical problem. In the longer term, the application seeks to exploit detailed structural knowledge for the synthesis of new ACE inhibitors with the aim of providing better drugs for the treatment of cardiovascular diseases in particular hypertension and fibrosis. A key feature will be the design of compounds that are specific for the N- or C-terminal domain of ACE with the expectation that this will provide selective compounds for therapy with fewer side effects.