A Cell Based HTS Approach for the Discovery of New Inhibitors of RSV

描述（由申请方提供）：目前，尚无市售疫苗可保护人类免受呼吸道合胞病毒（RSV）感染。RSV与大量发病率和死亡率相关，并且是婴儿和一岁以下儿童中细支气管炎和肺炎的最常见原因。由于人类对RSV感染没有免疫力，严重的下呼吸道疾病可能发生在任何年龄，特别是老年人或心脏、肺或免疫系统受损的人。用于急性感染的现有疗法是利巴韦林，其具有不一致的临床结果，以及预防性人源化单克隆抗体（来自MedImmune的Synagis），其昂贵且限于在高风险儿科患者中使用。由于住院和间接医疗费用，RSV感染的经济影响约为每年> 6.5亿美元。因此，迫切需要发现新的抗病毒药物来补充现有的化疗药物。高通量筛选（HTS）方法提供了在体外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孔基于细胞的测定法，该测定法使用基于发光的信号终点检测系统测量RSV感染在HEp-2细胞中诱导的CPE。这种分子HT分子筛选将为二次剂量反应试验提供基础，该试验将揭示活性化合物的关键抗病毒效力和细胞毒性信息，并为科学界提供通过体外评价大型化合物库快速鉴定RSV潜在抑制剂的试验。

DESCRIPTION (provided by applicant): Currently, there are no commercially available vaccines to protect humans against Respiratory syncytial virus (RSV) infection. RSV is associated with substantial morbidity and mortality and is the most common cause of bronchiolitis and pneumonia among infants and children under one year of age. Since humans have no immunity against RSV infection, severe lower respiratory tract disease may occur at any age, especially among the elderly or among those with compromised cardiac, pulmonary, or immune systems. The existing therapies for the acute infection are ribavirin, which has inconsistent clinical results, and the prophylactic humanized monoclonal antibody (Synagis(r) from MedImmune) that is expensive and limited to use in high risk pediatric patients. The economic impact of RSV infections due to hospitalizations and indirect medical costs is approximately > $ 650 million annually. Thus, there is a critical need to discover novel antiviral drugs to supplement existing chemotherapeutics. A high throughput screening (HTS) approach provides an opportunity to screen large compound libraries in vitro. We have developed and validated a 384-well cell-based assay that measures CPE induced in HEp-2 cells by RSV infection, using a luminescent-based detection system for signal endpoint. This molecular HT molecular screen will provide the foundation for secondary dose response assays that will divulge critical antiviral potency and cytotoxicity information on active compounds and bestow the scientific community with assays that allow for the rapid identification of potential inhibitors of RSV by evaluating large compound libraries in vitro.