Design & Synthesis of SARS Protease Inhibitors

该项目的主要目标是利用我们广泛的初步成果设计和开发SCLPro抑制剂，用于严重急性呼吸综合征(SARS)的治疗干预。自从2002年11月在广东省首次报告中国以来，SARS已经蔓延到其他亚洲国家、北美和欧洲。截至2003年7月，这种流行病已经影响了8000多名报告的个人，并导致774人死亡。最近发现的一种冠状病毒已被确定为SARS的病原体。到目前为止，还没有针对这种病毒的有效治疗方法。在病毒复制过程中，功能多肽 是由33.1-kD的HCoV 229E蛋白水解酶产生的，这种主要的蛋白水解酶被称为3C-like蛋白水解酶(SCLpro)，其活性对冠状病毒的复制至关重要。因此，这种蛋白酶被认为是治疗SARS和相关感染的药物开发的一个有吸引力的靶点。在前期工作中，我们根据SCLpro的X射线晶体结构、猪传染性胃肠炎病毒相关酶(TGEV MPRO)的X射线晶体结构和底物类似物六肽氯甲基酮(CMK)，设计合成了一系列SCLPro蛋白水解酶抑制剂。 抑制剂，以及来自SCLPro活性部位的模式化抑制剂AG-7088的结构信息。其中两种抑制剂不仅对SARS-SCLpro有相当的活性，而且在50-100微摩尔浓度下也抑制MHV(小鼠肝炎病毒)和SARS-HCoV细胞培养试验。我们还测定了这些与SARS-SCLpro络合的化合物的X-射线结构，分辨率为1.9埃。这项工作现在构成了我们拟议的研究的基础，在这些研究中，合成药物化学的力量将与结晶学和分子建模相结合，并被用来进一步开发具有临床潜力的新一代结构多样化和更有效的抑制剂。

The main objective of this project is to utilize our extensive preliminary results to design and develop SCLpro inhibitors for therapeutic intervention of Severe Acute Respiratory Syndrome (SARS). Since its first report in Guangdong Province, China, in November, 2002, SARS has spread to other Asian countries, North America and Europe. This epidemic already affected more than 8000 reported individuals by July, 2003, and resulted in 774 deaths. A recently discovered coronavirus has been identified as the etiological agent for SARS. Thus far, no effective therapy exists for this virus. During viral replication, the functional polypeptides are generated from polyproteins by proteolytic processing carried out by a 33.1-kD HCoV 229E main protease that is called 3C-like protease (SCLpro) and the activity of this SCLpro protease is critical to coronavirus replication. Thus, this protease is recognized as an attractive target for drug development for SARS and related infections. In preliminary work, we designed and synthesized a number of SCLpro protease inhibitors based upon the X-ray crystal structure of SCLpro, the X-ray crystal structure of related enzyme of porcine transmissible gastroenteritis (corona)virus (TGEV Mpro) with a substrate-analog hexapeptidyl chloromethyl ketone (CMK) inhibitor, as well as from structural information derived from amodeled inhibitor, AG-7088 in the SCLpro active site. Two of these inhibitors are not only quite active against SARS-SCLpro, but also inhibit MHV (mouse hepatitis virus) and SARS-HCoV cell culture assays at 50 to 100 micromolar concentrations. We have also determined the X-ray structures of these inhibitors complexed with SARS-SCLpro at a resolution of 1.9 Angstroms. This work now forms the basis of our proposed studies in which the power of synthetic medicinal chemistry will be combined with crystallography and molecular modeling and utilized to further develop a new generation of structurally diverse and more potent inhibitors with clinical potential.