Novel Inhibitors of Viral Replication

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。例如，流感病毒的NS 1蛋白，一种主要的毒力因子，抑制宿主基因表达和启动先天性和适应性免疫应答所需的信号转导。在感染的细胞中，NS 1定位于细胞核和细胞质中。基于NS 1在细胞核中作为基因表达抑制剂的功能的知识，我们对20万种合成化合物进行了高通量筛选。我们已经确定了八类新的化合物，显着恢复基因表达的存在下，NS 1，也显着抑制流感病毒复制和病毒介导的细胞死亡。这些化合物中的两种抑制流感病毒以外的病毒的复制，表明它们或者启动宿主抗病毒应答，或者抑制病毒复制所需的宿主因子。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这些化合物抑制病毒复制的机制，其双重目标是确定新的治疗靶点，并改善化合物的特性，从而可能导致新的分子治疗。我们将努力实现以下目标：目标1。研究抗病毒化合物的构效关系。UTSW合成化学实验室将根据我们最好的化合物产生靶向文库（约15-25种类似物），根据其抑制病毒复制的效力和缺乏细胞毒性进行优先排序。我们将进行定量结构活性分析，以建立每个化合物的药效团模型，以了解影响效力的关键因素。目标2.解决抗病毒化合物的ADME问题。我们将测试我们最好的化合物系列的溶解度，细胞吸收，代谢和细胞毒性。这一信息对于研究目标3中提出的流感病毒感染小鼠中的这些抑制剂将是重要的。我们将从纳摩尔范围内的活性化合物中制备亲和探针，用于靶向鉴定的生物化学方法。目标3：确定NS 1抑制剂对体内抗病毒应答的活性。我们将使用流感病毒感染的小鼠模型进行化合物预防感染功效的临床前试验。小鼠模型将用于测量化合物对肺中病毒复制和疾病严重程度的影响。还将评价干扰素应答。此外，将用VSV、牛痘病毒以及甲型和B型流感病毒感染小鼠，以研究化合物特异性。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。

DESCRIPTION (provided by applicant): Many viruses evade host defense mechanisms by targeting specific host vulnerabilities, revealing critical points in host pathways regulating antiviral responses. As an example, 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 in the cytoplasm. Based on the knowledge of NS1 functions in the nucleus as an inhibitor of gene expression, we performed a high throughput screen of 200,000 synthetic chemical compounds. We have identified eight classes of novel compounds that significantly restored gene expression in the presence of NS1 and also significantly inhibited both influenza virus replication and viral-mediated cell death. Two of these compounds inhibit the replication of viruses other than influenza virus, indicating that they either initiate a host antiviral response or inactivate a host factor required for virus replication. We will combine chemical biology, cell biology, and virology to investigate the mechanisms by which these compounds inhibit virus replication with the twin goals of identifying novel therapeutic targets and also improving the characteristic of the compounds as potential leads to novel molecular therapy. We will pursue the following aims: Aim 1. To investigate structure-activity relationship (SAR) of antiviral compounds. The UTSW Synthetic Chemistry Laboratory will produce targeted libraries (~15-25 analogs) based upon the best of our compounds, which are prioritized according to their potency to inhibit virus replication and their lack of cytotoxicity. We will conduct quantitative structure-activity analysis to establish a pharmacophore model of each compound, to provide an understanding of key elements affecting potency. Aim 2. To address issues of ADME of antiviral compounds. We will test our best compound series for, solubility, cellular absorption, metabolism and cellular toxicity. This information will be important to study these inhibitors in mice infected with influenza virus as proposed in Aim 3. We will make affinity probes from compounds that are active in the nanomolar range for a biochemical approach to target identification. Aim 3. To determine the activity of NS1 inhibitors on Antiviral Responses in vivo. We will use mice models of influenza virus infection for preclinical tests of compound efficacy in preventing infection. The mouse model will be used to measure the effects of compounds on virus replication in lungs and on the severity of disease. Interferon response will also be evaluated. In addition, mice will be infected with VSV, vaccina virus, and influenza A and B viruses to investigate compound specificity. 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.