High-throughput Screening for Inhibitors of Human Metapneumovirus

描述(申请人提供)：拟议研究的目标是确定人类偏肺病毒(HMPV)的小分子抑制剂。HMPV是全世界儿童下呼吸道感染的主要原因，并且在一生中反复感染。HMPV与老年人的严重疾病有关，住院率类似于流感， 以及患有哮喘、免疫损害和慢性肺部疾病等潜在疾病的人。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。我们开发了一种针对HMPV的荧光、基于细胞的高通量筛选(HTS)方法，该方法可以捕获除萌芽外病毒生命周期所有阶段的抑制物。我们对该分析进行了优化和验证，并在范德比尔特化学生物学研究所(VICB)的HTS设施中成功地对约3500种小化合物进行了HTS中试。我们建议使用从VICB的150,000个化合物库中选择的更大但有重点的化合物集来进行HMPV抑制剂的HTS。生物信息学工具将与药物化学合作者一起使用，以预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结果相关的结构。我们计划筛选75,000个化合物。我们将进行二次检测以确认HITS；测定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针对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工作精制化合物和进行临床前研究提供基础。该项目为针对一种重要的人类病原体的药物发现提供了一个独特的机会。所发现的化合物还将为病毒生物学提供强大的探针。

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.