Small Molecule Flavivirus Inhibitors

我们已经发现了新型的小分子化合物抑制黄病毒复制的药物。我们最有效的化合物有 50%有效浓度(EC50)为0.7微摩尔，50%细胞毒性浓度(CC50)为&gt；70 微摩尔，产生治疗(选择性)指数&gt；100。我们已经证明了这种化合物 保护动物免受西尼罗河病毒的致命感染，并将病毒RNA水平降低100倍以上 被感染的细胞；以及类似物抑制相关黄病毒的复制，如选择剂JAPAN 脑炎病毒(JEV)和A类生物防御病原体登革病毒(DENV)。的发展。 我们的小分子抗病毒药物将为治疗黄病毒感染提供一种新的方法，并将 补充疫苗接种方法。此外，因为很少有假定的黄病毒抑制剂被 如上所述，我们的工作对于建立理解结构-活性关系的基础是必不可少的 以及这些化合物如何影响宿主细胞功能和先天免疫。为了实现这些目标，我们 建议分析小分子化合物类似物的抗病毒活性和潜在毒性 以确定其抗病毒机制，并检测其对宿主细胞的影响。我们的特定 目标如下： 1.小分子抗黄病毒活性分析：黄病毒抑制剂及相关小文库 将制备类似物，并将合成对黄病毒株的细胞毒性和抗病毒作用 被量化。这些研究将确定抑制病毒所需的化学物质和菌株。 2.复合抑制机制的表征：病毒抑制的机制将是 特色化的。检测野生型感染和复制子表达的复合效应将有所帮助 描述抑制物的活性，以及对病毒蛋白活性的分析将有助于确定抑制机制。 对假定的耐药突变的分析将有助于评估抗药性是否是以合适的成本获得的 对病毒的作用，并将识别抑制剂靶向的基因。我们的结果将建立一种抗病毒作用模式， 以及优化抑制剂活性的方法。 3.对宿主细胞抑制作用的检测：虽然我们最有效的黄病毒抑制剂是无毒的， 它们对宿主细胞的具体影响几乎完全未知。为了填补这一空白，我们将监测化合物 对感染和未感染细胞的影响。这些结果将提供至关重要的数据，说明我们的新奇类 黄病毒抑制剂影响细胞表达谱和先天免疫应答机制

We have identified novel small compound inhibitors of flavivims replication. Our most potent compound has a 50% effective concentration (EC50) of 0.7 micromolar and a 50% cytotoxic concentration (CC50) of >70 micromolar, yielding a therapeutic (selectivity) index of >100. We have demonstrated that the compound protects animals from the lethality of WNV infection and reduces viral RNA levels over a hundred-fold in infected cells; and that analogues inhibit replication of related flaviviruses such as the select agent Japanese encephalitis virus (JEV) and the category A biodefense pathogen Dengue virus (DENV). The development of our small molecule antivirals will provide a new approach to the treatment of flavivirus infections, and will complement vaccine approaches. Moreover, because very few putative flavivirus inhibitors have been described, our work is essential to establish a foundation for understanding structure-activity relationships and how these compounds influence host cell functions and innate immunity. To achieve these goals, we propose to analyze the antiviral activities and potential toxicities of small compound analogues related to our inhibitors; to characterize their antiviral mechanisms; and to examine their effects on host cells. Our specific aims are as follows: 1 .Analysis of small molecule anti-flavivirus activities: Small libraries of flavivirus inhibitors and related analogues will be prepared, and compound cellular toxicities and antiviral effects against flavivirus strains will be quantitated. These studies will determine chemical and strain requirements for virus inhibition. 2. Characterization of the mechanisms of compound inhibition: The mechanisms of viral inhibition will be characterized. Examination of compound effects on wild type infections and replicon expression will help delineate inhibitor activities, and assays on viral protein activities will help define inhibition mechanisms. Analysis of putative drug-resistant mutants will help assess whether resistance is acquired at a fitness cost to the virus, and will identify genes targeted by inhibitors. Our results will establish a mode of antiviral action, and ways to optimize inhibitor activities. 3. Examination of inhibitor effects on host cells: Although our most effective flavivirus inhibitors are non-toxic, their specific effects on host cells are almost completely unknown. To fill this gap, we will monitor compound effects on infected and uninfected cells. These results will provide crucial data as to how our novel class of flavivirus inhibitors influence cellular expression profiles and innate immune response mechanisms