Discovery & Characterization of Filo-. Arena -and Alpha Virus Entry Inhibitors

发现

• 批准号: 7676506
• 负责人: ROBERT A DAVEY
• 金额: $ 15.21万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-15 至 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7676506
• 关键词: 2-cyclopentyl-5-(5-isoquinolylsulfonyl)-6-nitro-1H-benzo(D)imidazole; Address; Alphavirus; Animal Disease Models; Animal Model; Animals; Arenavirus; Basic Science; Biological Assay; Biological Warfare; Calcium; Calcium Channel Blockers; Calcium Signaling; Categories; Cell Communication; Cells; Cercopithecine Herpesvirus 1; Chemicals; Chikungunya virus; Clinic; Collaborations; Dependence; Disease; Dominant-Negative Mutation; Drug effect disorder; Ebola Hemorrhagic Fever; Ebola virus; Effectiveness; Emerging Communicable Diseases; FDA approved; Family; Filovirus; Frankfurt-Marburg Syndrome Virus; Genomics; Goals; Infection; Infection prevention; Junin virus; Libraries; Modeling; Modification; Pathogenesis; Pathway interactions; Pharmaceutical Preparations; Phosphotransferases; Publishing; RNA Viruses; Reporting; Role; Screening procedure; Series; Signal Transduction; Small Interfering RNA; Specificity; Structure-Activity Relationship; System; Technology; Testing; Therapeutic; Therapeutic Intervention; Translating; United States National Institutes of Health; Venezuelan Equine Encephalitis Virus; Venezuelan Equine Encephalomyelitis; Viral; Virus; Virus Diseases; Work; base; biodefense; chikungunya; comparative; density; dosage; drug candidate; drug mechanism; env Gene Products; follow-up; improved; inhibitor/antagonist; member; mouse model; novel therapeutics; overexpression; pathogen; prevent; receptor; release of sequestered calcium ion into cytoplasm; virus envelope

RNA viruses are prominent members of NIAID's Category A, B, and C lists of biodefense and emerging infectious disease agents. In this proposal therapeutics are being identified, characterized and developed. As our target we have chosen to focus on receptor interaction and entry. This is the first step in establishing an infection, and disruption has proved effective at preventing infection and virus spread. We have developed and published a series of assays that have become a platform technology for identifying such drugs and, importantly, for mechanism of action analysis. Our systems are now developed to the point where we can perform very high-density screens. To begin, we will use a library of 200,000 compounds in collaboration with the NIH Chemical Genomics Center. Active compounds will be prioritized based on comparative analyses of structure-activity relationships across the panel of assays determined from multiple dosages. Our UTMB team will work to develop hits into useful drugs. We will systematically compare two members each of three classes of virus, the filoviruses, Ebola and Marburg; arenaviruses, Lassa and Junin; and the alphaviruses, Venezuelan equine encephalitis and chikungunya viruses. Comparing each virus in one screen will be an effective means to identify potential drugs active against 1) specific virus types, 2) virus families, or 3) those of broad spectrum activity. All drugs will be tested against wild-type viruses. Next, we will follow up on our Ebola virus study, in which we identified FDA-approved drugs effective at blocking Ebola virus infection. In preliminary screens with small drug libraries and siRNA, we discovered that drugs that block calcium flux into cells are effective at inhibiting Ebola virus infection and cell entry, thereby preventing cytopathic effect. We will characterize each drug's mechanism of action. First, we will identify the most potent calcium channel blocking drugs via our specialized virus entry assays and use of wild-type virus. Second, we will address mechanism of action by identifying downstream signaling targets (e.g., CALM and CAM kinases) that may, in turn, be useful targets for anti-viral therapy. Third, we will examine the role of virus envelope protein in triggering this cascade by biochemically analyzing envelope protein-cell interactions. Finally, we will test the candidate drugs in animal disease models. We will also test other filoviruses, including Marburg virus. Our findings will aid in developing a robust filovirus therapy and an understanding of how calcium signaling functions in virus infection and potentially pathogenesis.

RNA病毒是NIAID的A、B和C类生物防御和新兴病毒列表中的突出成员。 传染病病原体。在这项提案中，正在确定、表征和开发治疗方法。 作为我们的目标，我们选择专注于受体相互作用和进入。这是第一步， 感染和破坏已被证明有效地防止感染和病毒传播。我们有 开发并发表了一系列分析，这些分析已经成为识别这种 药物，更重要的是，用于作用机制分析。我们的系统已经发展到 我们可以进行高密度的筛选。开始，我们将使用一个20万种化合物的库， 与NIH化学基因组学中心合作。活性化合物的优先顺序将根据 通过多项测定确定的一组测定的结构-活性关系的比较分析 剂量.我们的UTMB团队将致力于将热门产品开发成有用的药物。我们将系统地比较两个 三种病毒的成员，丝状病毒、埃博拉和马尔堡;沙粒病毒、拉沙病毒和朱宁; 以及甲病毒、委内瑞拉马脑炎和基孔肯雅病毒。比较每种病毒， 一种筛选将是鉴定对1）特定病毒类型，2）病毒 家庭，或3）广谱活性的那些。所有药物都将针对野生型病毒进行测试。接下来我们 我将继续我们的埃博拉病毒研究，在这项研究中，我们确定了FDA批准的有效阻断埃博拉病毒的药物， 病毒感染。在用小药物文库和siRNA进行的初步筛选中，我们发现， 阻断钙流入细胞可有效抑制埃博拉病毒感染和细胞进入， 致细胞病变效应我们将描述每种药物的作用机制。首先，我们将确定最 通过我们专门的病毒进入试验和野生型病毒的使用， 其次，我们将通过识别下游信号传导靶点（例如，平静和 CAM激酶），其又可以是抗病毒治疗的有用靶标。第三，我们将研究 病毒包膜蛋白在触发这一级联反应的生物化学分析包膜蛋白细胞 交互.最后，我们将在动物疾病模型中测试候选药物。我们还将测试其他 丝状病毒，包括马尔堡病毒。我们的研究结果将有助于开发一种强大的丝状病毒治疗方法和一种新的治疗方法。 了解钙信号在病毒感染中的作用和潜在的发病机制。