Using Chemical Biology to Interfere with the Influenza Virus Life Cycle

描述(申请人提供)：许多病毒通过针对特定的宿主漏洞来逃避宿主防御机制，揭示调节抗病毒反应的宿主途径的关键点。最近我们在了解流感病毒如何逃脱宿主对策方面的进展揭示了新的毒力机制，可以从药物上加以利用。流感病毒的NS1蛋白是一个主要的毒力因子，它抑制宿主基因的表达和信号转导，而宿主基因表达和信号转导是启动先天性和获得性免疫反应所必需的。在感染的细胞中，NS1定位于细胞核和细胞质。基于NS1作为基因表达抑制因子的功能，我们对20万个合成化合物进行了高通量筛选，并发现了新的NS1抑制剂。我们选择了八类新化合物，它们在NS1存在的情况下显著恢复了基因表达，并同时抑制了流感病毒的复制和宿主细胞的死亡。我们认为，NS1的核活性和/或细胞质活性的拮抗剂将成为病毒复制和致病的新型抑制剂。我们将结合化学生物学、细胞生物学和病毒学来研究我们鉴定的化合物抑制流感病毒复制的机制。我们将描述两个小分子家族，它们可以作为分子治疗的先导。此外，拟议的研究将为流感病毒逃避宿主防御途径的机制提供新的见解，这些途径可以进一步在药理学上进行靶向。我们将追求以下目标：目的1.研究NS1抑制剂的构效关系。目的2.测定NS1抑制剂的体内抗病毒活性。目的3.确定抑制NS1功能的化合物的靶向途径和作用机制。总而言之，这些研究可能会揭示抗病毒治疗的新线索，并提供关于病毒-宿主相互作用和途径的新机制的信息。 公共卫生相关性：需要新的疗法来预防人类的病毒疾病，因为病毒对当前使用的药物产生抗药性，而且很难通过接种疫苗来保护整个人口。该项目将开发能够抑制流感病毒和其他病毒生长的新型化合物，达到在动物模型中证明它们可以预防病毒疾病的阶段，这是朝着开发新的人类抗病毒疗法迈出的第一步。

DESCRIPTION (provided by applicant): Many viruses evade host defense mechanisms by targeting specific host vulnerabilities, revealing critical points in host pathways regulating antiviral responses. Recent advances in our understanding of how influenza virus escapes host countermeasures have exposed novel mechanisms of virulence that can be exploited pharmacologically. The NS1 protein of influenza virus, a major virulence factor, inhibits host gene expression and signal transduction required to mount innate and adaptive immune responses. In infected cells, NS1 is localized in the nucleus and the cytoplasm. Based on the function of NS1 as inhibitor of gene expression, we performed a high throughput screen (HTS) of 200,000 synthetic chemical compounds and identified novel inhibitors of NS1. We selected eight classes of novel compounds that significantly restored gene expression in the presence of NS1, and that also inhibited both influenza virus replication and host cell death. We propose that antagonists of either nuclear and/or cytoplasmic activities of NS1 will be novel inhibitors of viral replication and pathogenesis. We will combine chemical biology, cell biology, and virology to investigate the mechanisms by which the compounds we have identified inhibit influenza virus replication. We will characterize two families of small molecules that can serve as leads for molecular therapy. In addition, the proposed studies will provide new insights into mechanisms by which influenza virus evades host defense pathways that can be further targeted pharmacologically. We will pursue the following aims: Aim 1. To investigate structure-activity relationship (SAR) of NS1 inhibitors. Aim 2. To determine the activity of NS1 inhibitors on Antiviral Responses in vivo. Aim 3. To identify targeted pathways and mechanisms of action of compounds which inhibit NS1 function. In sum, these studies will likely reveal novel leads for antiviral therapies as well as provide information on novel mechanisms of viral-host interactions and pathways. PUBLIC HEALTH RELEVANCE: Novel therapeutics to prevent viral disease in humans are needed because viruses become resistant to currently used drugs and it is difficult to protect the entire population by vaccination. This project will develop new classes of chemical compounds capable of inhibiting the growth of influenza virus and other viruses to the stage where they can be shown to prevent viral disease in animal models, the first step towards developing leads to new antiviral therapeutics for humans.