Identifying inhibitors of West Nile and dengue viruses

鉴定西尼罗河病毒和登革热病毒的抑制剂

• 批准号: 7118877
• 负责人: PEI-YONG SHI
• 金额: $ 0.93万
• 依托单位: WADSWORTH CENTER
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-06-01 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7118877
• 关键词: West Nile virus; antiviral agents; bioassay; biotechnology; cell line; chemical registry /resource; cooperative study; dengue; dengue virus; drug discovery /isolation; drug screening /evaluation; emerging infectious disease; genetic manipulation; high throughput technology; pharmacokinetics; reporter genes; technology /technique development; tissue /cell culture; virus diseases; virus genetics; virus replication

DESCRIPTION (provided by applicant): West Nile (WN) and dengue (DEN) viruses are emerging flaviviruses and potential biodefense pathogens that cause significant human diseases. Neither vaccine nor effective chemotherapy is currently available for WN and DEN infections in humans. The objective of this project is to identify novel inhibitors of WN and DEN viruses. We will accomplish the following two interim objectives. (1) Develop novel high-throughput assays to screen for inhibitors of WN and DEN replication. Using subgenomic WN and DEN replicons expressing reporter genes, we will develop reporting cell lines that can be used for high-throughput screening of inhibitors against all targets involved in viral replication. As a proof-of-principle, we have already established such a reporting cell line for high-throughput screening of WN inhibitors. We will develop a similar high-throughput assay for anti-DEN drug discovery. (2) Identify new classes of inhibitors of WN and DEN and analyze their modes of action. In collaboration with ViroPharma, Inc., we will use the high-throughput assays to identify new classes of WN and DEN inhibitors through screening a large number of compound libraries. We will study the potential inhibitors by (i) examining whether the compounds can inhibit other members of the flaviviruses, and (ii) analyzing the modes of action of the compounds through both biochemical (enzyme assays) and genetic approaches (selection of compound-resistant viruses). The high-throughput assay-approach proposed in this application has been proven to be fruitful in antiviral drug discovery (e.g. hepatitis C virus). At the completion of these studies, we expect to have identified novel inhibitors of WN and DEN, and to have determined the modes of action of these inhibitors. These results will be significant, because they will form a foundation for development of an efficacious chemotherapy of WN and DEN infections. The reporting cell lines developed in this project will also be useful for many aspects of flavivirus research such as studying genome packaging and viral particle formation.

描述（由申请人提供）：西尼罗河（WN）和登革热（DEN）病毒是新兴的黄病毒和潜在的生物防御病原体，可引起严重的人类疾病。目前尚无针对人类 WN 和 DEN 感染的疫苗和有效的化疗。该项目的目标是鉴定 WN 和 DEN 病毒的新型抑制剂。我们将实现以下两个中期目标。 (1) 开发新的高通量测定法来筛选 WN 和 DEN 复制的抑制剂。使用表达报告基因的亚基因组 WN 和 DEN 复制子，我们将开发可用于高通量筛选针对病毒复制涉及的所有靶标的抑制剂的报告细胞系。作为原理验证，我们已经建立了这样一个用于高通量筛选 WN 抑制剂的报告细胞系。我们将开发一种类似的高通量检测方法来发现抗 DEN 药物。 (2) 鉴定新一类WN和DEN抑制剂并分析其作用方式。我们将与 ViroPharma, Inc. 合作，利用高通量检测通过筛选大量化合物库来鉴定新类别的 WN 和 DEN 抑制剂。我们将通过（i）检查这些化合物是否可以抑制黄病毒的其他成员，以及（ii）通过生化（酶测定）和遗传方法（选择化合物抗性）分析化合物的作用模式来研究潜在的抑制剂病毒）。本申请中提出的高通量测定方法已被证明在抗病毒药物（例如丙型肝炎病毒）的发现方面卓有成效。在完成这些研究后，我们期望能够鉴定出 WN 和 DEN 的新型抑制剂，并确定这些抑制剂的作用模式。这些结果意义重大，因为它们将为开发 WN 和 DEN 感染的有效化疗奠定基础。该项目开发的报告细胞系也可用于黄病毒研究的许多方面，例如研究基因组包装和病毒颗粒形成。