Novel Inhibitors of Viral Replication

病毒复制的新型抑制剂

• 批准号: 7939350
• 负责人: Beatriz MA Fontoura
• 金额: $ 41.91万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7939350
• 关键词: Address; Affect; Affinity; Animal Model; Antisense Oligonucleotides; Antiviral Agents; Antiviral Response; Antiviral Therapy; Biochemical; Biology; Cell Death; Cell Nucleus; Cells; Cellular biology; Cercopithecine Herpesvirus 1; Characteristics; Chemicals; Cytoplasm; Drug usage; Elements; Gene Expression; Goals; Growth; Host Defense Mechanism; Human; Immune response; Infection prevention; Integration Host Factors; Interferons; Knowledge; Laboratories; Libraries; Lung; Measures; Mediating; Metabolism; Modeling; Molecular; Mus; PTPN11 gene; Pathway interactions; Population; Preclinical Testing; Resistance; Series; Severity of illness; Signal Transduction; Solubility; Specificity; Staging; Structure; Structure-Activity Relationship; Sum; Synthesis Chemistry; Testing; Therapeutic; Toxic effect; Twin Multiple Birth; Vaccination; Vaccinia virus; Vesicular stomatitis Indiana virus; Viral; Virulence Factors; Virus; Virus Diseases; Virus Inhibitors; Virus Replication; absorption; analog; base; cytotoxicity; high throughput screening; improved; in vivo; influenza virus INS1 protein; influenzavirus; inhibitor/antagonist; mouse model; new therapeutic target; novel; novel therapeutics; overexpression; pharmacophore; prevent; response; virology

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.

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。例如，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启动先天性和获得性免疫反应所需的信号转导。在感染的细胞中，NS1定位于细胞核和细胞质中。基于NS1在核中作为基因表达抑制因子的功能的知识，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化合物，它们在NS1存在的情况下显著恢复了基因表达，并显著抑制了流感病毒复制和病毒介导的细胞死亡。其中两种化合物抑制流感病毒以外的病毒复制，表明它们要么启动宿主抗病毒反应，要么使病毒复制所需的宿主因子失活。我们将结合化学生物学、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，同时发现新的治疗靶点，并改善化合物的特性，因为这可能导致新的分子治疗。我们将追求以下目标：目的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生产目标文库(~15-25个类似物)，这些化合物根据其抑制病毒复制的效力和无细胞毒性而优先考虑。我们将进行定量的结构-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。目的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性、细胞吸收、新陈代谢和细胞毒性。这些信息对于在目标3中建议的在感染流感病毒的小鼠身上研究这些抑制物将是重要的。我们将从活性在纳摩尔范围内的化合物中制作亲和探针，用于生化方法识别靶标。目的3.测定NS1抑制剂的体内抗病毒活性。我们将使用流感病毒感染的小鼠模型进行预防感染的复合疗效的临床前测试。老鼠模型将被用来测量化合物对肺部病毒复制和疾病严重程度的影响。干扰素反应也将进行评估。此外，小鼠还将感染VSV、痘苗病毒以及甲型和乙型流感病毒，以研究化合物的特异性。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。