Structure Based Design of Dengue Virus Fusion Inhibitors

基于结构的登革热病毒融合抑制剂设计

• 批准号: 7176132
• 负责人: Sherri Zimmerman Millis
• 金额: $ 29.03万
• 依托单位: PANTHERA BIOPHARMA, LLC
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-02-15 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7176132
• 关键词: Antiviral Agents; Back; Binding; Biochemical; Biological Assay; C-terminal; Categories; Cell membrane; Cells; Chemical Structure; Class; Condition; Culicidae; DNA Sequence Rearrangement; Dengue; Dengue Hemorrhagic Fever; Dengue Shock Syndrome; Dengue Virus; Development; Disease; Drug Delivery Systems; Family; Flavivirus; Flavivirus Infections; Fluorescence; Fluorescence Polarization; Geographic Distribution; Goals; Hawaii; Human; In Vitro; Incidence; Infection; Knowledge; Label; Laboratories; Life; Life Cycle Stages; Ligand Binding; Measures; Mediating; Membrane; Membrane Fusion; Membrane Lipids; Methods; Modeling; Molecular Target; National Institute of Allergy and Infectious Disease; Peptides; Pharmaceutical Preparations; Phase; Population; Positioning Attribute; Preclinical Drug Evaluation; Process; Proteins; Research Project Grants; Risk; Screening procedure; Serotyping; Site; Staging; Structure; Testing; Vaccines; Viral; Viral Fusion Proteins; Virion; Virus; Virus Diseases; Virus Replication; Work; base; burden of illness; design; dimer; drug discovery; env Gene Products; inhibitor/antagonist; insight; medical schools; member; molecular modeling; novel; pathogen; prevent; programs; small molecule libraries; stem; three dimensional structure; virtual

DESCRIPTION (provided by applicant): Over half of the world population is at risk for infection by dengue virus, a mosquito borne member of the Flavivirus family that consists of four distinct serotypes. Approximately 50 to 100 million infections occur annually resulting in an estimated 500,000 cases of life threatening dengue hemorrhagic fever or dengue shock syndrome. Due to the increased incidence and spreading geographic distribution of dengue infection, it is considered to be an emerging disease and is identified by NIAID as a category A priority pathogen. Despite the significant disease burden associated with dengue infection there is presently no approved vaccine or antiviral drug. The goal of this project is to discover and develop antiviral drugs that inhibit dengue virus infection by blocking the cell entry step in the virus life cycle. Dengue infection of host cells is mediated by the envelope protein, a class II viral fusion protein responsible for virus attachment and for triggering fusion between the virus and host cell membranes. Hawaii Biotech has expressed the envelope proteins of dengue virus in purified native form. The 3-dimensional structures of the dengue-2 envelope protein in both a pre and a post fusion state have recently been solved as a collaborative project with Stephen Harrison's laboratory at Harvard Medical School. The structures reveal two potential drug target sites on the envelope protein for inhibitors of the fusion process. One target is a hydrophobic pocket present in the native envelope of the virus particle, and the other is a channel present in the fusion intermediate form of the protein involved in the late stages of membrane fusion. These targets will be examined by molecular modeling through a process of virtual drug screening to find potential inhibitor compounds that will then be tested experimentally. The purified envelope protein undergoes the conformational rearrangements of the fusion process in vitro under appropriate conditions. This will form the basis for development of fluorescence based assay methods that can be used to identify fusion inhibitors. A cell based assay for dengue envelope mediated membrane fusion will also be developed for testing of inhibitor candidates. By evaluating two distinct molecular targets that represent different stages in the fusion process, and by exploring several different assay designs, we intend to be in a position at the conclusion of Phase I to select the best path to discovery of effective antiviral drugs for dengue and other Flaviviruses during Phase II.

描述（由申请人提供）：超过一半的世界人口有感染登革热病毒的风险，登革热病毒是由四种不同血清型组成的黄病毒家族的蚊媒成员。每年发生大约5000万至1亿例感染，导致估计500，000例危及生命的登革出血热或登革休克综合征。由于登革热感染的发病率增加和地理分布的扩大，它被认为是一种新出现的疾病，并被NIAID确定为A类优先病原体。尽管与登革热感染相关的重大疾病负担，但目前还没有批准的疫苗或抗病毒药物。该项目的目标是发现和开发通过阻断病毒生命周期中的细胞进入步骤来抑制登革热病毒感染的抗病毒药物。宿主细胞的登革感染由包膜蛋白介导，包膜蛋白是一种II类病毒融合蛋白，负责病毒附着和触发病毒与宿主细胞膜之间的融合。夏威夷生物技术公司已经以纯化的天然形式表达了登革病毒的包膜蛋白。作为与哈佛医学院的Stephen Harrison实验室的合作项目，登革热2型包膜蛋白在融合前和融合后状态下的三维结构最近已经得到解决。结构揭示了两个潜在的药物靶位点的包膜蛋白的融合过程的抑制剂。一个靶点是存在于病毒颗粒的天然包膜中的疏水口袋，另一个是存在于膜融合后期阶段所涉及的蛋白质的融合中间体形式中的通道。这些目标将通过虚拟药物筛选过程的分子建模进行检查，以找到潜在的抑制剂化合物，然后进行实验测试。纯化的包膜蛋白在适当的条件下进行体外融合过程的构象重排。这将形成开发可用于鉴定融合抑制剂的基于荧光的测定方法的基础。还将开发一种用于登革热包膜介导的膜融合的基于细胞的测定法，用于测试抑制剂候选物。通过评估代表融合过程中不同阶段的两种不同分子靶标，并通过探索几种不同的检测设计，我们打算在I期结束时选择最佳途径，以在II期发现登革热和其他黄病毒的有效抗病毒药物。