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）设计和优化了一种高通量的RABV检测方法，从而能够初步筛选狂犬病抑制剂。筛选了大约22，000种化合物，并确定了数百种命中物。使用HepG 2毒性测定和GFP正交测定进一步分类命中分子。在该试验组中具有理想特征的小分子被进一步用于病毒和机制试验中的进一步表征。 被鉴定为对RABV有效的化合物的子集先前已经被推进到用于其他疾病/适应症的临床试验，并且具有有利的生物学和PK特征。这些化合物将作为药物再利用研究的基础。我们最近将其中几种化合物推进到初步的小鼠PK研究中，随后在CDC的狂犬病仓鼠模型中进行了疗效研究。该系列的作用机制也正在研究中。从HTS努力的新命中被用于定量结构活性关系研究（QSAR），以确定其他活性化合物和化学型。这些结果将激发药物化学运动，我们寻求开发新的小分子治疗RABV的治疗。

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.