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）：针对流感病毒感染的广谱疗法是解决流感大流行问题的迫切需要，流感大流行是全球公共卫生的主要威胁。干扰病毒进入是一种新颖而有吸引力的控制病毒感染的治疗策略。HIV抑制剂恩福韦肽（T-20）证明了这种方法的原理。我们的目标是发现能抑制H5N1禽流感和其他潜在大流行性流感病毒进入的非肽小分子。我们的策略是靶向包膜糖蛋白血凝素（HA），它介导流感病毒通过受体结合和与宿主细胞融合进入。透明质酸是一类融合蛋白就像HIV的Gp120和副粘病毒的F蛋白一样。一类病毒融合蛋白在融合过程中经历一系列构象重排，形成融合发夹结构。在融合过程中，这种结构促进了病毒和细胞包膜的并置，并由蛋白质和蛋白质的相互作用维持。副粘病毒的小分子进入抑制剂已被确定干扰这种融合发夹结构的形成。由于类似的结构存在于HA和其他I类融合蛋白中，因此结果支持了我们将HA作为药物发现策略的假设。针对这一保守位点的抑制剂将对多种亚型有效，包括一种新出现的大流行毒株。我们将使用一种表达HA （H5亚型）的伪病毒作为替代模型，模拟HA介导的进入，并在BSL2条件下筛选进入抑制剂。在初步研究中，我们开发了一种敏感的伪病毒检测方法，用于筛选血凝素抑制剂。在这个STTR项目的第一阶段，我们将优化该检测方法，以快速筛选结构多样的小分子文库。我们的合作者、伊利诺伊大学芝加哥分校的Lijun Rong博士将以盲法方式对hit进行确认，并由西奈山医学院的Adolfo Garcia-Sastre博士在一个增强型BSL 3实验室对其抗H5禽流感病毒的抗流感活性进行评估。确认的命中将根据其与其他亚型的频谱进行评估，并根据其作用机制确定优先级。这些新型流感疗法将阻断病毒进入并抑制病毒与细胞接触产生的细胞毒性。由于保守结构域的微小偏差会阻止病毒与内体膜的融合，因此预计它们的耐药发生率也很低。在第二阶段，我们将通过合理的药物设计计划来推进最有希望的支架，并将在动物模型中测试先导化合物的功效和毒性。最活跃的化合物，毒性最小，将推进到IND的研究。公共卫生相关性：流感是一种高度传染性的急性呼吸道疾病，其特点是每年复发性流行和周期性的全球大流行。疫苗是目前预防流感病毒感染的主要策略，但只有在与流行病毒类型相匹配的情况下才有效，而且不能预先开发出针对新出现的大流行毒株的疫苗。我们的目标是开发一种抗流感治疗药物，通过靶向包膜蛋白血凝素（HA）的保守融合和受体结合域来阻止病毒进入，并将对所有亚型有效，包括新出现的大流行毒株。