Chemical intervention of influenza virus RNA nuclear export

 描述（由申请方提供）：流感病毒在美国每年造成约36，000例死亡，在全球每年造成约500，000例死亡。致病性极强的菌株导致全球大量死亡，例如1918年的大流行导致全球约3000万人死亡。目前，只有两种方法可用于预防或治疗流行性和大流行性流感，疫苗接种和病毒复制抑制剂。尽管疫苗接种对同源毒株非常有效，但在老年人中失去了效力，并且受到维鲁的高度可变性的限制，迫使每年重新配制疫苗以匹配当前流感病毒流行毒株的抗原性。许多药物已被批准用于治疗流感。这些药物抑制病毒脱壳或病毒传播，但这些相对少量的抗病毒药物的使用受到耐药病毒株出现的限制。因此，显然需要用于治疗流感病毒疾病的其他治疗方式。在这个应用中，我们处理一个新的发现平台的开发，用于识别抑制流感病毒RNA核输出的化合物。流感病毒RNA被导入宿主细胞核进行复制，然后作为mRNA或vRNA从细胞核输出，分别表达病毒蛋白或产生新的病毒颗粒。因此，细胞核中vRNA或病毒mRNA的阻断导致病毒复制的抑制。我们有证据表明，我们可以在完整的感染细胞中跟踪单个病毒vRNA和mRNA片段，并且这个过程可以被调节。我们将使用该测定法，使用我们的~ 200，000个化合物文库和我们的高通量成像平台来检测病毒RNA核输出的抑制。该屏幕适用于应用程序的R21阶段。对于R33相，将所选择的命中物进行构效关系，以改进 在基于细胞和基于动物的模型中，用各种流感病毒株进行的效力、选择性、稳定性和毒性特征。总之，通过该发现平台鉴定的化合物将可能代表一类新的潜在抗病毒药物，其可用于对抗流感病毒和可能使用类似核输出机制的其他病毒。

 DESCRIPTION (provided by applicant): Influenza viruses cause approximately 36,000 deaths annually in the United States and ~500,000 deaths worldwide per year. Strains that are extremely pathogenic have been responsible for high numbers of deaths worldwide, such as the 1918 pandemic which led to ~30 million deaths around the world. Currently, there are only two approaches available for preventing or treating epidemic and pandemic influenza, vaccination and inhibitors of viral replication. Vaccination, although highly effective against homologous strains, looses efficacy in the elderly, and it is limited by the highly mutable nature of the viru, forcing the annual reformulation of the vaccine to match the antigenicity of the current influenza virus circulating strains. A number of drugs have been approved for the treatment of influenza. These drugs inhibit virus uncoating or virus spread but the use of these relatively small number of antiviral drugs is limited by the appearance of resistant virus strains. Thus, there is a clear need for additional therapeutic modalities for the treatment of influenza virus disease. In this application, we deal with the development of a new discovery platform for the identification of chemical compounds that inhibit influenza virus RNA nuclear export. Influenza virus RNAs are imported into the host cell nucleus for replication and are then exported from the nucleus as mRNAs or vRNAs to express viral proteins or generate new viral particles, respectively. Therefore, blockage of vRNAs or viral mRNAs in the nucleus results in inhibition of virus replication. We have evidence demonstrating that we can follow single viral vRNA and mRNA segments in intact infected cell and that this process can be regulated. We will use this assay to detect inhibition of viral RNA nuclear export using our ~200,000 compound library and our high throughput imaging platform. The screen is proposed for the R21 phase of the application. For the R33 phase, the selected hits will be subjected to structure activity relationship for improving potency, selectivity, stability and toxicity profile in cell based and animal based models with various strains of influenza virus. In sum, the identified compounds by this discovery platform will likely represent a new class of potential antivirals that could be useful for antagonizing influenza virus and possibly other viruses that use a similar nuclear export machinery.