Development of MS2045 for inhibition of Zika methyltransferase

项目摘要 寨卡病毒(ZIKV)是黄病毒属的成员，包括其他节肢动物传播的人类病原体 如登革热病毒和西尼罗河病毒等。ZIKV与新生儿小头畸形症的联系以及 成人的吉兰-巴雷综合征加强了开发疫苗的措施以及开发疫苗的努力 以ZIKV生命周期和生存中心的酶活性为靶点的抗病毒药物。这样的一个 酶活性由甲基转移酶(MTase)结构域编码，位于N端 非结构蛋白NS5。采取结构知情的方法，我们成功地确定了一部小说 “类铅”化合物(MS2045)，用于抑制ZIKV的NS5 MTase活性并阻止其复制。 MS2045为进一步的化学和开发更有效的抑制剂提供了基础。在目标1中， 我们将a)设计MS2045类似物，具有与特定氨基酸建立特定相互作用的能力 ZIKV MTase作为提供对人RNA5‘-帽MTase的额外选择性的手段；b)化学 合成这些类似物并将它们生产到95%的纯度，用于体外和基于细胞的分析，以及用于 结构研究。在目标2中，我们将a)进行生物物理分析，以评估这些类似物的能力 与人RNA5‘-帽MTase相比，选择性结合ZIKV NS5-MTase，并检测其结合能力 抑制RNA甲基化；b)在基于病毒细胞的检测中测试这些类似物，以评估它们阻断病毒的有效性 复制；c)使用精选类似物确定结构ZIKV NS5-MTase，以进行额外的迭代循环 结构活性关系(SARS)。总的来说，这些研究将有助于识别MS2045的类似物 可以潜在地发展成为来自ZIKV(和其他致病病毒)的NS5-MTase的有效和选择性抑制剂 黄病毒)

Project Summary The Zika virus (ZIKV) is a member of the Flavivirus genus that includes other arthropod-borne human pathogens such as dengue virus and West Nile virus, among others. ZIKV’s link to microcephaly in newborn infants and the Guillan-Barré syndrome in adults has invigorated measures to develop a vaccine, as well as efforts to develop antivirals based on targeting enzymatic activities central to the life cycle and survival of ZIKV. One such enzymatic activity is encoded by the methyltransferase (MTase) domain, located at the N-terminus of the nonstructural protein NS5. Taking a structure informed approach, we have succeeded in identifying a novel “lead-like” compound (MS2045) for the inhibition of ZIKV’s NS5 MTase activity and for blocking its replication. MS2045 provides a basis for further chemistry and the development of even more potent inhibitors. In aim 1, we will a) design MS2045 analogs with the capacity to establish specific interactions with unique amino acids of ZIKV MTase as a means to provide additional selectivity against the human RNA 5’-cap MTases; b) chemically synthesize these analogs and produce them to a purity of 95% for in vitro and cell-based assays, and for structural studies. In aim 2, we will a) perform biophysical assays to assess the ability of these analogs to selectively bind the ZIKV NS5-MTase as compared to the human RNA 5’-cap MTases and test their ability to inhibit RNA methylation; b) test these analogs in viral cell-based assays to assess their efficacy in blocking viral replication; c) determine structures ZIKV NS5-Mtase with select analogs for additional, iterative rounds of structure activity relationships (SARs). Collectively, these studies will help to identify analogs of MS2045 that can be potentially developed into potent and selective inhibitors of NS5-MTase from ZIKV (and other pathogenic flaviviruses)