Identification of novel influenza antivirals

说明(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，它们每年仍造成约3.6万人死亡和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。两类药物已被批准用于预防和治疗流感。令人震惊的是，过去十年出现了抗药性以及最近大流行性(H1N1)和高致病性(H5N1)甲型流感病毒株的暴发，最好的情况是，作为治疗其他病毒疾病的方法，将使用联合药物疗法来提供最有效的预防和治疗，并抑制出现更多的抗药性。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，以至于金刚烷胺已经变得无效。目前迫切需要新的、更有效的治疗策略。在这里，我们提供了一种创新的方法来确定甲型流感病毒RNA聚合酶PA和PB1亚单位组装的新抑制剂。这些亚基通过短螺旋片段的堆积在高度保守的相互作用部位结合。有趣的是，通过突变或与多肽抑制PA-PB1的结合阻止了病毒的复制和共晶结构，表明这种结合应该服从小分子抑制剂。我们发展了一种新的基于细胞和荧光的PA-PB1亚单位结合分析方法，用于高通量筛选化学文库和确认有效的HIT化合物。使用这种方法，我们建议鉴定作用于新的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。 与公共卫生相关：在过去十年中，出现了一些抗药性和/或高致病性的流感变种，大大增加了季节性和大流行性流感感染的潜在影响。这项提议详细说明了一种创新的方法，以确定急需的新的抑制物和潜在的治疗方法来预防和治疗甲型流感感染。

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.