qHTS to Identify Inhibitors of Rabies Virus

在此期间，NCGC和合作团队使用两种表达荧光素酶和GFP报告基因的重组狂犬病病毒(ERA)设计和优化了高通量顺应性RABV检测，使狂犬病抑制剂的初步筛选成为可能。大约22,000种化合物被筛选出来，并确定了数百个匹配项。HIT分子进一步用HepG2毒性试验和GFP正交试验进行筛选。在这个分析小组中，具有理想轮廓的小分子被提出用于病毒和机械分析的进一步表征。 一组被确定为对RABV有效的化合物子集此前已进入其他疾病/适应症的临床试验，并具有良好的生物学和PK特征。这些化合物将作为药物再用途研究的基础。我们最近已经将其中几种化合物推进到初步的小鼠PK研究中，随后在疾控中心的狂犬病仓鼠模型中进行疗效研究。这一系列药物的作用机制也在研究中。HTS工作中的新命中正被用于定量结构活性关系研究(QSAR)，以确定更多的活性化合物和化学类型。这些结果将激发一场药物化学运动，在那里我们寻求开发治疗RABV的新型小分子疗法。

During this period, NCGC and the collaborative team worked to design and optimize a high-throughput amenable RABV assay using two recombinant rabies viruses (ERA) expressing luciferase and GFP reporters, enabling primary screening for Rabies inhibitors. Approximately 22,000 compounds were screened and several hundred hits were identified. Hit molecules were further triaged using a HepG2 toxicity assay and a GFP orthogonal assay. Small molecules with ideal profiles in this assay panel were advanced for further characterization in viral and mechanistic assays. A subset of compounds identified as efficacious against RABV have previously been advanced to clinical trials for other diseases/indications and have a favorable biological and PK profile.These compounds will serve as the basis for a drug repurposing study. We have recently advanced several of these compounds to preliminary mouse PK studies, followed by efficacy studies in a rabies hamster model at the CDC. The mechanism of action for this series is also being investigated. Novel hits from the HTS efforts are being used in a quantitative structure activity relationship study (QSAR) to identify additional active compounds and chemotypes. These results will inspire a medicinal chemistry campaign, where we seek to develop novel small molecule therapeutics for RABV treatment.