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NMR-based discovery of influenza hemagglutinin probes

基于核磁共振的流感血凝素探针的发现

基本信息

批准号：
8500191
负责人：
Michael S. Caffrey
金额：
$ 18.74万
依托单位：
UNIVERSITY OF ILLINOIS AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-07-01 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8500191
关键词：
Affinity Antiviral Agents Binding Binding Sites Biological Assay Cells Cessation of life Chemicals Development Disease Outbreaks Drug resistance Endocytosis Endosomes Exhibits Experimental Designs FDA approved Future GlaxoSmithKline brand of zanamivir Goals H5 hemagglutinin HIV Health Hemagglutinin Human Influenza Influenza A Virus, H1N1 Subtype Influenza A Virus, H5N1 Subtype Influenza Hemagglutinin Libraries Mediating Membrane Membrane Fusion Membrane Proteins Methods Molecular Conformation Molecular Weight Monoclonal Antibodies Mutagenesis NMR Spectroscopy NOESY Neuraminidase Oseltamivir Peptides Pharmaceutical Preparations Pharmacotherapy Play Proteins Protocols documentation Recombinants Resistance Role SARS coronavirus Site Structure Techniques Therapeutic Therapeutic Intervention Viral Viral Envelope Proteins Virus Work analog base design env Gene Products follow-up high reward high risk improved influenza outbreak influenzavirus inhibitor/antagonist innovation insight novel pandemic disease pathogen receptor receptor binding receptor mediated endocytosis research study resistant strain screening sialic acid receptor small molecule small molecule libraries therapeutic development zanamivir

项目摘要

DESCRIPTION (provided by applicant): There are more than 250,000 influenza-related deaths each year. Of special concern are the influenza H1N1 and H5N1 strains, which harbor H1 and H5 hemagglutinin (HA) and present potential for future pandemic outbreaks. Current treatments include Tamiflu (oseltamivir) and Relenza (zanamivir), which target the membrane protein neuraminidase (NA). Unfortunately, resistance is increasing in circulating influenza strains. For example, the 2008-2009 H1N1 strain exhibited ~100% resistance against Tamiflu, and thus influenza may be considered as a drug-resistant pathogen. As a consequence, new targets in influenza are urgently needed. HA plays a critical role in influenza entry and thus it i an attractive and novel target for therapeutic intervention. In the first step of influenza entry, A binds to the cellular receptor, sialic acid. Subsequently, the virus enters the cell via receptor-mediated endocytosis. In the endosome, the resulting low pH triggers a large conformational change in HA, which subsequently causes the viral membrane to fuse with the endosomal membrane to allow escape from the endosome. Entry inhibitors could conceivably target either the receptor binding step (binding inhibitors) or the membrane fusion step (fusion inhibitors). In this project we will develop NMR-based screening methods of small molecule libraries to influenza H1 and H5 HA with the long-term goal of developing binding and fusion inhibitors. The Specific Aims are: (1) discovery of chemical probes that bind to influenza H1 and H5 HA in the neutral pH conformation; (2) discovery of chemical probes that bind to influenza H1 and H5 HA in the low pH conformation; (3) identify the site of probe binding by NMR competition experiments. Importantly, the NMR methods could be exploited to identify and improve small molecule therapeutics, characterize probe-envelope interactions in whole virus, and be applied to other viruses such as Ebola, HIV and SARS-CoV.

描述（由申请人提供）：每年有超过 250,000 人死于流感。特别值得关注的是 H1N1 和 H5N1 流感病毒株，它们含有 H1 和 H5 血凝素 (HA)，并有可能在未来爆发大流行。目前的治疗方法包括达菲（奥司他韦）和瑞乐沙（扎那米韦），其靶向膜蛋白神经氨酸酶（NA）。不幸的是，流行的流感病毒株的耐药性正在增加。例如，2008-2009 年 H1N1 病毒株对达菲表现出约 100% 的耐药性，因此流感可被视为耐药病原体。因此，迫切需要流感的新靶点。 HA 在流感进入过程中发挥着关键作用，因此它是治疗干预的一个有吸引力且新颖的目标。在流感进入的第一步，A 与细胞受体唾液酸结合。随后，病毒通过受体介导的内吞作用进入细胞。在内体中，由此产生的低pH值会引发HA的巨大构象变化，随后导致病毒膜与内体膜融合，从而从内体中逃逸。可以想象，进入抑制剂可以靶向受体结合步骤（结合抑制剂）或膜融合步骤（融合抑制剂）。在这个项目中，我们将开发基于 NMR 的 H1 和 H5 流感 HA 小分子文库筛选方法，长期目标是开发结合和融合抑制剂。具体目标是：（1）发现在中性pH构象下与H1和H5流感HA结合的化学探针； (2) 发现在低pH构象下与流感H1和H5 HA结合的化学探针； (3)通过NMR竞争实验鉴定探针结合位点。重要的是，核磁共振方法可用于识别和改进小分子疗法，表征整个病毒中的探针-包膜相互作用，并应用于其他病毒，如埃博拉病毒、HIV 和 SARS-CoV。