Identification of novel influenza antivirals

描述（由申请人提供）：甲型流感病毒是全球最大的人类健康风险之一。尽管疫苗对季节性流感感染提供了重要的保护，但仅在美国，每年仍有36000人死亡，20万人住院。此外，疫苗的开发、生产和分发所涉及的固有时间限制了它们对迅速出现的疫情的潜在效力。已有两类药物被批准用于预防和治疗流感。令人震惊的是，过去十年出现了耐药性，最近爆发了甲型流感大流行（H1N1）和高致病性（H5N1）菌株。在治疗其他病毒性疾病时，最好采用联合药物疗法，以提供最有效的预防和治疗，并抑制其他耐药性的出现。然而，目前只有一类药物（神经氨酸酶抑制剂）可用；金刚烷胺耐药性已经变得如此普遍，金刚烷胺已经失效。目前迫切需要新的和更有效的治疗策略。在这里，我们提供了一种创新的方法来鉴定流感A RNA聚合酶亚基PA和PB1组装的新抑制剂。这些亚基通过短螺旋段的包装在高度保守的相互作用位点结合。有趣的是，通过诱变或多肽抑制PA-PB1结合可阻断病毒复制和共晶结构，这表明这种结合应该适用于小分子抑制剂。我们已经开发了一种新的基于细胞和荧光的PA-PB1亚基关联分析，用于化学文库的高通量筛选和虚拟命中化合物的确认。利用这种方法，我们建议鉴定作用于新型甲型流感靶点的抑制化合物，以开发新的广谱流感药物治疗。

DESCRIPTION (provided by applicant): The influenza A virus represents one of the greatest global human health risks. Although vaccines provide significant protection from seasonal flu infections they still account for an estimated 36,000 deaths and 200,000 hospitalizations per year in the US alone. Furthermore, the inherent time involved in development, production and distribution of vaccines limits their potential efficacy against rapidly emerging outbreaks. Two classes of drugs have been approved for influenza prophylaxis and treatment. Alarmingly, the past decade has witnessed the emergence of drug resistance as well as recent outbreaks of pandemic (H1N1) and highly pathogenic (H5N1) strains of influenza A. Optimally, as adopted for the treatment of other viral diseases, combination drug therapies would be used to provide the most effective prophylaxis and treatment and to inhibit the emergence of additional drug- resistances. However, only one class of drug (neuraminidase inhibitors) is currently available; amantadine-resistance has become so widespread the amantadanes have become ineffective. There is currently an urgent need for new and more effective therapeutic strategies. Here we provide an innovative approach to identify novel inhibitors of the assembly of the influenza A RNA polymerase subunits PA and PB1. These subunits associate at highly conserved interaction sites through packing of short helical segments. Interestingly, inhibiting PA-PB1 association by mutagenesis or with peptides blocks viral replication and co-crystal structure indicates the binding should be amenable to small- molecule inhibitors. We have developed a novel cell and fluorescence-based PA-PB1 subunit association assay for high-throughput-screening of chemical libraries and confirmation of virtual hit compounds. Using this approach we propose to identify inhibitory compounds acting at a novel influenza A target site for the development of new broad-spectrum influenza drug therapies. PUBLIC HEALTH RELEVANCE: Over the past decade the emergence of a number of drug-resistant and/or highly pathogenic variants of influenza have dramatically increased the potential impact of both seasonal and pandemic influenza infection. This proposal details an innovative approach to identify much-needed novel inhibitors and potential therapeutics for the prevention and treatment of influenza A infection.