Identification and characterization of novel drugs that target the Influenza

该项目的总体目标是确定针对流感病毒复制的新疗法。每年流感大流行造成的全球健康负担，加上高致病性流感病毒株的出现，突显了对新的有效治疗方法的迫切需要。目前用于治疗流感的药物(金刚烷胺和神经氨酸酶抑制剂)的主要担忧是抗药性突变的发展，从而否定了治疗效果。已发表的证据表明，针对流感病毒RNA依赖的RNA聚合酶(RdRp)是一种合理的抗病毒治疗方法。RdRp负责许多功能，包括5‘帽识别、核酸内切酶活性、复制、转录和多聚腺苷酸化。最近，冷冻-EM重建研究发现分支核糖核蛋白(RNPs)结构是假定的复制中间产物，并提出了一种通过RNP模板上的第二聚合酶活性来实现病毒复制的机制。第二个聚合酶活性被认为是聚合酶复合体的功能。显然，RdRp提供了多种功能结构域，可以成为抗病毒药物治疗的靶点。以往的研究表明，聚合酶复合体PB1亚单位保守区的突变会产生失活的RNA聚合酶。我们假设，专门针对聚合酶复合体的化合物可能会减少逃逸突变的频率，或者促进不适合复制的逃逸突变。我们最近从以前的HTS筛查中发现了潜在的热门化合物，这些化合物在RdRp瞬时试验中显著抑制流感病毒聚合酶的活性。在CPE试验中，这些化合物对三种不同的流感病毒株都有效。我们建议对这些化合物进行表征，并使用新型小分子文库的高通量筛选(HTS)来识别抗聚合酶候选化合物，对它们进行化学优化，并在流感小鼠模型中建立它们的有效性。 \

The overall goal of this project is to identify new therapies that target influenza virus replication. The global health burden of annual influenza epidemics coupled with the emergence of highly pathogenic strains of influenza virus has highlighted the urgent need for new effective treatments A primary concern with the current drugs (amantadines and neuraminidase inhibitors) used to treat influenza is the development of resistance mutations that negate therapeutic benefit. Published evidence suggests that targeting the influenza virus RNA dependent RNA polymerase (RdRp) is a rational approach for antiviral therapy. The RdRp is responsible for a number of functions including 5'cap recognition, endonuclease activity, replication, transcription, and polyadenylation. Recently, cryo-EM reconstitution studies identified branchedribonucleoproteins (RNPs) structures as putative replication interhiediates and suggested a mechanism for viral replication by a second polymerase activity on the RNP template . The second polymerase activity is believed to be a function ofthe polymerase complex. Clearly, the RdRp provides multiple functional domains that could be targets for antiviral drug therapy. Previous studies showed that mutations in the conserved regions of PB1 subunit of the polymerase complex produce inactive RNA polymerase. We hypothesize that compounds that specifically target the polymerase complex might reduce the frequency of escape mutations, or promote escape mutants that are unfit for replication. We have recently identified potential hit compounds from previous HTS screens that significantly inhibit the influenza virus polymerase activity in an RdRp transient assay. These hit compounds were effective against three different strains of influenza viruses in CPE assays. We propose to characterize these compounds and use high-throughput screening (HTS) of novel small molecule libraries to identify anti-polymerase candidate compounds, chemically optimize them, and establish their effectiveness in an influenza mouse model. \