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）：急需针对流感病毒感染的广谱治疗剂来解决流感大流行问题，这是对全球公共卫生的主要威胁。干扰病毒进入是控制病毒感染的一种新颖而有吸引力的治疗策略。这种方法的原理证据来自HIV抑制剂恩夫韦肽（T-20）。我们的目标是发现能够抑制禽流感H5 N1和其他潜在大流行性流感病毒进入的非肽类小分子。我们的策略是靶向包膜糖蛋白血凝素（HA），HA通过受体结合和与宿主细胞融合介导流感病毒进入。HA是I类融合蛋白，类似于HIV Gp 120和副粘病毒的F蛋白。I类病毒融合蛋白在融合过程中经历一系列构象重排，导致融合发夹结构。这种所得结构促进融合过程中病毒和细胞包膜的并置，并通过蛋白质-蛋白质相互作用维持。已经鉴定了干扰这种融合发夹结构形成的副粘病毒的小分子进入抑制剂。由于类似的结构存在于HA和其他I类融合蛋白，结果支持我们的假设，靶向HA作为药物发现的策略。针对该保守位点的抑制剂将对多种亚型具有活性，包括新出现的大流行毒株。我们将使用一种表达HA（H5亚型）的假型病毒，该病毒已被开发为替代模型，以模拟HA介导的进入并在BSL 2条件下筛选进入抑制剂。在初步研究中，我们已经开发和表征了一种灵敏的假型病毒检测用于筛选HA抑制剂。在这个STTR项目的第一阶段，我们将优化快速筛选结构多样性小分子的大型（> 100，000）文库的检测方法。我们的合作者、伊利诺伊大学芝加哥分校的Lijun Rong博士将以盲法确认命中，西奈山医学院的Adolfo Garcia-Sastre博士将在增强的BSL 3实验室中评估它们对活H5禽流感病毒的抗流感活性。将根据其他亚型评价确认的命中的谱，并根据其作用机制进行优先排序。这些新型流感治疗剂将阻断病毒进入并抑制由病毒-细胞接触引起的细胞毒性。由于保守结构域的微小偏离将阻止病毒与内体膜融合，因此预期其耐药性发生率也较低。在第二阶段，我们将通过合理的药物设计计划开发最有前途的支架，并将在动物模型中测试先导化合物的功效和毒性。活性最高、毒性最小的化合物将进入IND启动研究。公共卫生相关性：流感是一种传染性极强的急性呼吸道疾病，其特征是每年反复流行，并在世界范围内定期发生大流行。疫苗是目前预防流感病毒感染的主要策略，只有当它们与流行的病毒类型相匹配时才有效，并且不能针对新出现的大流行毒株提前开发。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结构域来防止病毒进入，并对所有亚型都有活性，包括新出现的大流行毒株。