Using Chemical Biology to Interfere with the Influenza Virus Life Cycle

描述（由申请人提供）：许多病毒通过靶向特定宿主漏洞来逃避宿主防御机制，揭示了调节抗病毒反应的宿主途径中的关键点。我们对流感病毒如何逃脱宿主对策的理解的最新进展已经暴露了可以被重新利用的新的毒力机制。流感病毒的NS 1蛋白是一种主要的毒力因子，可抑制宿主基因表达和信号转导，而这些基因表达和信号转导是启动先天性和适应性免疫应答所必需的。在感染的细胞中，NS 1定位于细胞核和细胞质中。基于NS 1作为基因表达抑制剂的功能，我们对20万种合成化合物进行了高通量筛选（HTS），并鉴定了新型NS 1抑制剂。我们选择了八类新型化合物，它们在NS 1存在下显著恢复了基因表达，并且还抑制了流感病毒复制和宿主细胞死亡。我们建议，无论是核和/或细胞质活性的NS 1拮抗剂将是新的病毒复制和发病机制的抑制剂。我们将结合联合收割机化学生物学、细胞生物学和病毒学来研究我们所鉴定的化合物抑制流感病毒复制的机制。我们将描述两个家庭的小分子，可以作为分子治疗的线索。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，这些机制可以进一步靶向流感病毒。我们将努力实现以下目标：目标1。研究NS 1抑制剂的构效关系。目标2.确定NS 1抑制剂对体内抗病毒应答的活性。目标3：确定抑制NS 1功能的化合物的靶向途径和作用机制。总之，这些研究将可能揭示抗病毒治疗的新线索，并提供有关病毒-宿主相互作用和途径的新机制的信息。 公共卫生相关性：需要预防人类病毒性疾病的新疗法，因为病毒对目前使用的药物产生耐药性，并且难以通过疫苗接种保护整个人群。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，使其能够在动物模型中预防病毒性疾病，这是开发新的人类抗病毒疗法的第一步。

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.