A Cell Based HTS Approach for the Discovery of New Inhibitors of the H5N1 Virus

基于细胞的 HTS 方法用于发现 H5N1 病毒新抑制剂

• 批准号: 7305167
• 负责人: WILLIAM E SEVERSON
• 金额: $ 2.5万
• 依托单位: SOUTHERN RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-06-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7305167
• 关键词: Africa; Antiviral Agents; Asia; Avian Influenza; Biological Assay; Canis familiaris; Cells; Centers for Disease Control and Prevention (U.S.); Communities; Detection; Dose; Economics; End Point; Europe; Human; Immunity; In Vitro; Influenza A Virus, H5N1 Subtype; Kidney; Libraries; MDCK cell; Measures; Molecular; Molecular Bank; Molecular Probes; Principal Investigator; Research; Resources; Screening procedure; Signal Transduction; System; United States; Vaccines; Virus; Virus Diseases; Virus Replication; base; cost; follow-up; high throughput screening; influenzavirus; inhibitor/antagonist; killings; novel; pandemic influenza; programs; response; scaffold; vaccine development

DESCRIPTION (provided by applicant): Currently, there is no commercially available vaccine to protect humans against the highly pathogenic avian influenza H5N1 virus that is spreading across Asia, Europe, and Africa. Since humans have no immunity against any H5 viruses, the Centers for Disease Control and Prevention estimates that the economic impact of the next influenza pandemic could cost the United States billions of dollars, devastate the world economy and potentially kill one billion people worldwide. Thus, there is a critical need for antiviral drugs to supplement vaccine development and existing chemotherapeutics. A high throughput screening (HTS) approach provides an opportunity to screen large compound libraries. We have developed and validated a 384-well cell-based assay that measures CPE induced in Madin Darby canine kidney (MDCK) cells by influenza virus infection, using a luminescent-based detection system for signal endpoint. This molecular screen will provide the scientific community with an assay that allows for the rapid identification of potential inhibitors of influenza virus by evaluating large compound libraries in vitro. The proposed studies aim to gain access to the Molecular Libraries Screening Center Network (MLSCN) high throughput screening resources to facilitate the discovery of new molecular probes for the inhibition of avian influenza strain H5N1 virus. To achieve the objective of this proposal, the specific aim is to assist the MLSCN in transferring the validated HTS assay to the appropriate screening center and provide technical support for follow up confirmatory assays on single dose 'hits' from the primary assay in a HTS dose response format.

描述（由申请人提供）：目前，没有市售疫苗可保护人类免受正在亚洲、欧洲和非洲蔓延的高致病性禽流感H5N1病毒的侵害。由于人类对任何H5病毒都没有免疫力，疾病控制和预防中心估计，下一次流感大流行的经济影响可能会使美国损失数十亿美元，破坏世界经济，并可能导致全球10亿人死亡。因此，迫切需要抗病毒药物来补充疫苗开发和现有的化疗药物。高通量筛选（HTS）方法提供了筛选大型化合物文库的机会。我们已经开发并验证了一种384孔细胞检测法，该方法使用基于发光的信号终点检测系统，测量流感病毒感染在Madin达比犬肾（MDCK）细胞中诱导的CPE。这种分子筛选将为科学界提供一种检测方法，通过体外评估大型化合物库，快速鉴定流感病毒的潜在抑制剂。该研究旨在获得分子库筛选中心网络（MLSCN）的高通量筛选资源，以促进新的抑制禽流感病毒株H5N1的分子探针的发现。为实现本提案的目标，具体目标是协助MLSCN将经验证的HTS检测转移至适当的筛查中心，并以HTS剂量响应格式为初步检测的单次剂量“命中”的后续确证性检测提供技术支持。