High-throughput Screening for Inhibitors of Human Metapneumovirus

描述（由申请方提供）：拟定研究的目的是鉴定人偏肺病毒（HMPV）的小分子抑制剂。HMPV是全球儿童下呼吸道感染的主要原因，并且在整个生命中发生复发性感染。HMPV与老年人的严重疾病相关，住院率与流感相似， 以及患有潜在疾病如哮喘、免疫功能低下和慢性肺病的人。目前还没有获得许可的HMPV疫苗或药物，因此对抗病毒药物的需求尚未得到满足。我们开发了一种荧光，基于细胞的高通量筛选（HTS）检测HMPV，捕获抑制剂的病毒生命周期的所有阶段，除了出芽。我们优化并验证了该测定，并在范德比尔特化学生物学研究所（VICB）HTS设施中对约3500种小化合物进行了成功的试点HTS。我们建议使用从超过150，000种化合物的VICB库中选择的更大但集中的化合物集对HMPV抑制剂进行HTS。生物信息学工具将与药物化学合作者一起使用，以预先筛选和排除已知的有毒化合物，并丰富与我们的试点筛选中发现的命中相关的结构。我们计划筛选75,000种化合物。我们将进行二次试验以确认命中;确定IC50，CC50和选择性指数;并确定抑制剂的作用机制。这些实验的结果将确定HMPV和其他呼吸道病毒的新的潜在抗病毒药物。此外，这些发现将为SAR工作提供基础，以改进化合物并进行临床前研究。该项目为针对重要人类病原体的药物发现提供了独特的机会。发现的化合物也将为病毒生物学提供强有力的探针。

DESCRIPTION (provided by applicant): The goal of the proposed research is to identify small molecule inhibitors of human metapneumovirus (HMPV). HMPV is a leading cause of lower respiratory infection in children worldwide, and recurrent infections occur throughout life. HMPV is associated with severe illness in older adults, with hospitalization rates similar to influenza, and persons with underlying conditions such as asthma, immune compromise, and chronic pulmonary disease. There are no licensed vaccines or drugs for HMPV and thus there is an unmet need for antivirals. We developed a fluorescent, cell-based high-throughput screening (HTS) assay for HMPV that captures inhibitors of all stages of the viral lifecycle except budding. We optimized and validated the assay, and performed a successful pilot HTS against ~3500 small compounds in the Vanderbilt Institute of Chemical Biology (VICB) HTS facility. We propose to conduct a HTS for HMPV inhibitors using a larger but focused compound set selected from the VICB library of >150,000 compounds. Bioinformatics tools will be used in conjunction with medicinal chemistry collaborators to prescreen and exclude known toxic compounds, and to enrich for structures related to the hits discovered in our pilot screen. We plan to screen 75,000 compounds. We will perform secondary assays to confirm hits; determine IC50, CC50, and selectivity index; and define mechanisms of action of inhibitors. The results of these experiments will identify new potential antivirals for HMPV and other respiratory viruses. Further, the findings will provide a foundation for SAR work to refine compounds and conduct preclinical studies. This project offers a unique opportunity for drug discovery against an important human pathogen. The compounds discovered will also provide powerful probes of viral biology.