Identification of Novel Broad Spectrum Influenza Virus Inhibitors

新型广谱流感病毒抑制剂的鉴定

• 批准号: 7482015
• 负责人: Arnab Basu
• 金额: $ 18.92万
• 依托单位: MICROBIOTIX, INC
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-15 至 2010-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7482015
• 关键词: Acute; Address; Animal Model; Antiviral Agents; Appearance; Asia; Avian Influenza; Avian Influenza A Virus; Binding Sites; Biological Assay; Birds; Cell-Mediated Cytolysis; Cells; Chicago; Chimeric Proteins; Class; Condition; DNA Sequence Rearrangement; Drug Design; Epidemic; Europe; Gastrointestinal tract structure; Geographic Distribution; Glycoproteins; Goals; H5 hemagglutinin; H5 influenza virus; HIV; HIV Envelope Protein gp120; Helix (Snails); Hemagglutinin; Human; Human Virus; Illinois; Immunity; Incidence; Infection; Influenza; Influenza A Virus, H5N1 Subtype; Influenza Hemagglutinin; Influenza Therapeutic; Ion Channel; Laboratories; Lead; Libraries; Life; Luciferases; Lung diseases; Mediating; Medical; Membrane; Minor; Modeling; N-terminal; Neuraminidase inhibitor; Paramyxovirus; Peptides; Pharmaceutical Preparations; Phase; Population; Probability; Production; Proteins; Public Health; Recurrence; Reporter; Resistance; Resistance development; Risk; Route; Screening procedure; Series; Signal Transduction; Site; Small Business Technology Transfer Research; Specificity; Structure; System; T-20; Testing; Therapeutic; TimeLine; Toxic effect; Universities; Vaccines; Variant; Viral; Virus; Virus Diseases; Virus Inhibitors; absorption; anti-influenza; anti-influenza drug; base; biosafety level 2 facility; blind; college; cost; cytotoxic; cytotoxicity; day; drug discovery; ear helix; env Gene Products; falls; flu activity; high throughput screening; indexing; influenza virus strain; influenzavirus; inhibitor/antagonist; ion channel blocker; novel; novel therapeutics; pandemic disease; pandemic influenza; prevent; programs; protein protein interaction; receptor; receptor binding; scaffold; small molecule

DESCRIPTION (provided by applicant): Broad-spectrum therapeutics against influenza virus infections are critically needed to address the problem of influenza pandemics, a major threat to the public health globally. Interfering with virus entry is a novel and attractive therapeutic strategy to control virus infection. Proof of principle of this approach has come from the HIV inhibitor enfuvirtide (T-20). Our goal is to discover nonpeptidic small molecules that will inhibit entry of avian influenza H5N1 and other potentially pandemic influenza viruses. Our strategy is to target envelope glycoprotein hemagglutinin (HA), which mediates influenza virus entry through receptor binding and fusion with host cells. HA is class I fusion protein like the HIV Gp120 and F protein of paramyxoviruses. The class I virus fusion proteins undergo a series of conformational rearrangements during fusion that leads to fusion hairpin structure. This resultant structure promotes the juxtaposition of the viral and cellular envelopes during fusion and is sustained by protein protein interactions. Small molecule entry inhibitors of paramyxoviruses have been identified that interfere with the formation of this fusion hairpin structure. Because analogous structures are present in HA and other class I fusion proteins, the results support our hypothesis of targeting HA as a strategy for drug discovery. Inhibitors targeting this conserved site will be active against multiple subtypes, including a newly emerged pandemic strain. We will use a pseudotype virus expressing HA (H5 subtype), which has been developed as a surrogate model, to mimic HA mediated entry and screen for entry inhibitors under BSL2 conditions. In preliminary studies, we have developed and characterized a sensitive pseudotype virus assay for screening HA inhibitors. In Phase I of this STTR project, we will optimize the assay for rapid screening of a large (>100,000) library of structurally diverse small molecules. Hits will be confirmed in a blind fashion by our collaborator Dr Lijun Rong, University of Illinois at Chicago and will be evaluated for their anti-influenza activity against live H5 avian influenza viruses in an enhanced BSL 3 laboratory by Dr Adolfo Garcia-Sastre, at Mount Sinai Medical College. Confirmed hits will be evaluated for their spectrum against other subtypes and prioritized based on their mechanism of action. These novel influenza therapeutics will block virus entry and suppress cellular cytotoxicity resulting from virus-cell contact. They will also be expected to have a low incidence of resistance development since minor deviations in the conserved domains would prevent fusion of virus with endosomal membrane. In Phase II, we will progress the most promising scaffolds through a rational drug design program, and will test lead compounds for efficacy and toxicity in animal models. The most active compound, with the least toxicity, will advance to IND enabling studies. PUBLIC HEALTH RELEVANCE: Influenza is a highly infectious acute respiratory disease, characterized by recurrent annual epidemics and periodic major worldwide pandemics. Vaccines, currently the primary strategy for protection against influenza virus infection, are only effective if they match the circulating virus type(s) and cannot be developed in advance against new emerging pandemic strain(s). Our goal is to develop an anti-influenza therapeutic that will prevent virus entry by targeting the conserved fusion and receptor binding domains of envelope protein hemagglutinin (HA), and will be active against all subtypes, including a newly emerging pandemic strain.

描述（由申请人提供）：迫切需要针对流感病毒感染的广谱治疗剂来解决全球公共卫生的主要威胁，以解决流感大流感的问题。干扰病毒进入是一种新颖而有吸引力的治疗策略，可控制病毒感染。这种方法的原理证明来自HIV抑制剂Enfuvirtide（T-20）。我们的目标是发现非肽的小分子，这些分子将抑制禽流感H5N1和其他潜在的流血流感病毒的进入。我们的策略是靶向包膜糖蛋白血凝蛋白（HA），该糖蛋白（HA）通过受体结合和与宿主细胞融合来介导流感病毒的进入。 HA是I类融合蛋白，例如HIV GP120和Faryxovirus的F蛋白。 I类病毒融合蛋白在融合过程中会经历一系列构象重排，从而导致融合发夹结构。这种结果结构促进了融合过程中病毒和细胞信封的并置，并由蛋白质蛋白质相互作用维持。已经确定了小分子的小分子进入抑制剂，这些抑制剂会干扰这种融合发夹结构的形成。由于HA和其他I类融合蛋白中存在类似结构，因此结果支持我们将HA作为药物发现策略的假设。针对该保守位点的抑制剂将对多种亚型（包括新出现的大流行菌株）有效。我们将使用表达HA（H5亚型）的伪型病毒（已开发为替代模型）来模仿HA介导的进入和在BSL2条件下进入抑制剂的筛选。在初步研究中，我们已经开发并表征了一种敏感的假型病毒测定法，用于筛选HA抑制剂。在此STTR项目的第一阶段，我们将优化用于快速筛选结构上小分子的大型（> 100,000）库的测定法。我们的合作者Lijun Rong博士，伊利诺伊大学的芝加哥大学将以盲目的方式确认HITS，并将在Sinai Mount Medical College的Adolfo Garcia-Sastre Dr Adolfo Garcia-Sastre的增强的BSL 3实验室中对其针对Live H5 Avian流感病毒的反激动人物活动进行评估。确认的命中将对其他亚型的频谱进行评估，并根据其作用机理优先级。这些新型的流感疗法将阻止病毒进入并抑制病毒细胞接触导致的细胞细胞毒性。由于保守域中的较小偏差会阻止病毒与内体膜融合，因此他们还将有预期的耐药性发展率低。在第二阶段，我们将通过一个合理的药物设计程序来进步最有希望的脚手架，并将测试铅化合物在动物模型中的疗效和毒性。最活跃的化合物，最少的毒性将促进研究。公共卫生相关性：流感是一种高度感染性的急性呼吸道疾病，其特征是年度流行病和周期性的主要全球大流行病。目前，疫苗是针对流感病毒感染的保护的主要策略，只有在与循环病毒类型相匹配的情况下才有效，并且不能预先针对新的新兴大流行菌株（S）开发。我们的目标是开发一种抗炎治疗，该治疗方法将通过靶向包膜蛋白血凝素（HA）的保守融合和受体结合结构域，以防止病毒进入，并将针对所有亚型（包括新出现的大流行病）进行活性。