Inhibition of the Membrane Fusion Proteins of Flaviviruses and Alphaviruses

黄病毒和甲病毒膜融合蛋白的抑制

• 批准号: 7255226
• 负责人: MARGARET KIELIAN
• 金额: $ 24.9万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-05-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7255226
• 关键词: Alphavirus; Animals; Antiviral Agents; Antiviral Therapy; Binding; Biological Assay; Categories; Cells; Chimeric Proteins; Class; Co-Immunoprecipitations; Collaborations; Complication; Country; Culicidae; Dengue; Dengue Hemorrhagic Fever; Dengue Virus; Developed Countries; Developing Countries; Dominant-Negative Mutation; Encephalitis; Flavivirus; Goals; Health; Helix (Snails); Human; In Vitro; Infection; Institution; Japanese Encephalitis; Lead; Mediating; Membrane; Membrane Fusion; Methods; Molecular Conformation; Numbers; Pathway interactions; Peptide Library; Peptides; Protein Fragment; Proteins; Reaction; Research; Screening procedure; Semliki forest virus; Structure; Testing; Ticks; Transmembrane Domain; United States National Institutes of Health; Viral Hemorrhagic Fevers; Virus; Virus Diseases; Virus Inhibitors; West Nile virus; Western Equine Encephalitis Virus; Yellow fever virus; base; biodefense; dimer; ear helix; high throughput screening; in vitro Assay; inhibitor/antagonist; member; novel; pathogen; protein protein interaction; small molecule; small molecule libraries; tool; trimer core; vector

DESCRIPTION (provided by applicant): Flaviviruses and alphaviruses are spread by mosquito and tick vectors and cause severe human and animal illnesses such as encephalitis and hemorrhagic fever. These viruses include many potential bioterrorist agents that are category A-C pathogens, such as the flaviviruses dengue, West Nile, Japanese encephalitis and yellow fever viruses, and the alphaviruses Venezuelan, eastern, and western equine encephalitis viruses. Dengue virus (DV) is currently of particular concern as it has dramatically reemerged to become endemic in more than 100 countries including the US, and is now a global health problem. There are an estimated 50-100 million cases of dengue fever and 500,000 cases of the more lethal complication dengue hemorrhagic fever per year, with significant impact on both human health and the economies of developing countries. Antiviral strategies for the flaviviruses and alphaviruses are urgently needed. The flavivirus and alphavirus membrane fusion proteins are members of the class II virus fusion proteins. They are structurally very similar and refold to a homotrimer form to mediate virus fusion and infection. In collaboration with Dr. F¿lix Rey, we have recently determined the structure of the homotrimer conformation of the fusion protein from the alphavirus Semliki Forest virus (SFV). The SFV homotrimer structure is strikingly similar to that of DV. Using the structure as a guide, we have developed protein fragments that act as specific dominant-negative inhibitors of SFV and DV fusion and infection. Based on this information, we now plan to develop a general screen for inhibitors of class II fusion reactions. In aim 1 we will establish and optimize in vitro methods to follow the protein-protein interactions during the class II fusion protein refolding reaction. In aim 2, we will adapt this assay to a high throughput format, and use it to screen peptide and small molecule libraries available at our institution and through the NIH and Northeast Biodefense Consortium. Ultimately, such inhibitors will be lead compounds for antiviral therapy, and important research tools to understand the class II virus fusion reaction. Flaviviruses and alphaviruses include many important human pathogens and potential bioterrorist threats such as the flaviviruses dengue, West Nile, Japanese encephalitis and yellow fever viruses, and the alphaviruses Venezuelan, eastern, and western equine encephalitis viruses. Dengue virus is currently of particular concern as it has dramatically reemerged to become endemic in more than 100 countries including the US, and is now a global health problem. This application focuses on developing new antiviral strategies for the flaviviruses and alphaviruses, based on blocking the activity of the proteins involved in the initial entry of the virus into the cell.

描述（由申请人提供）：黄病毒和甲病毒通过蚊子和蜱虫传播，引起严重的人类和动物疾病，例如脑炎和出血热。这些病毒包括许多潜在的生物恐怖分子，属于 A-C 类病原体，例如黄病毒登革热、西尼罗河病毒、日本脑炎病毒和黄热病病毒，以及委内瑞拉、东部和西部马脑炎病毒甲病毒。目前，登革热 (DV) 已在包括美国在内的 100 多个国家急剧流行，成为全球性健康问题。估计有 50-1 亿例登革热病例和 50 万例登革热病例。每年有更多的致命并发症登革出血热，对人类健康和发展中国家的经济产生重大影响，迫切需要针对黄病毒和甲病毒的抗病毒策略。黄病毒和甲病毒膜融合蛋白是 II 类病毒融合蛋白的成员，它们在结构上非常相似，并重新折叠成同源三聚体形式，以介导病毒融合和感染。 lix Rey，我们最近确定了甲病毒塞姆利基森林病毒（SFV）的融合蛋白的同源三聚体结构。SFV同源三聚体结构与DV的结构非常相似，我们以该结构为指导开发了蛋白质。作为 SFV 和 DV 融合和感染的特异性显性失活抑制剂的片段 基于此信息，我们现在计划开发 II 类融合反应抑制剂的通用筛选。优化体外方法以跟踪 II 类融合蛋白重折叠反应过程中的蛋白质-蛋白质相互作用。在目标 2 中，我们将采用高通量格式，并使用它来筛选我们机构可用的肽和小分子文库。最终，此类抑制剂将成为抗病毒治疗的先导化合物，以及了解包括许多重要人类病原体在内的 II 类病毒融合反应的重要研究工具。生物恐怖威胁，例如黄病毒登革热病毒、西尼罗河病毒、日本脑炎病毒和黄热病病毒，以及甲病毒委内瑞拉、东部和西部马脑炎病毒，目前特别令人担忧，因为它已在 100 多个国家中急剧流行。包括美国在内的国家，目前已成为一个全球性健康问题。该应用的重点是基于阻断蛋白质的活性，开发针对黄病毒和甲病毒的新抗病毒策略。参与病毒最初进入细胞。