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Inhibiting protein-primed replication of human enteroviruses.

抑制人类肠道病毒的蛋白质引发复制。

基本信息

批准号：
8725044
负责人：
CATHERINE H SCHEIN
金额：
$ 20.28万
依托单位：
FOUNDATION FOR APPLIED MOLECULAR EVOLUTN
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-01 至 2016-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8725044
关键词：
Acids Acute Adverse effects Antiviral Agents Binding Binding Sites Bioinformatics Biological Assay Capsid Cardiac Myocytes Cardiomyopathies Cell Culture Techniques Cells Centers for Disease Control and Prevention (U.S.)Chronic Complex Computer Simulation Consensus Coupling Coxsackie Viruses Crystallization Culture Media Data Dilated Cardiomyopathy Diversity Library Docking Drug Design Echo Viruses Enterovirus Enterovirus Infections Enzymes Family Picornaviridae Gastrointestinal tract structure Goals Head Heart Heart Transplantation Housing Human Human poliovirus Immune system In Vitro Infection Insulin-Dependent Diabetes Mellitus Lead Libraries Mammalian Cell Methods Modeling Mus Myocarditis Nebraska Online Systems Organ Organic Synthesis Pancreas Peptides Pharmaceutical Preparations Pharmacologic Substance Pharmacology Phase Play Poliomyelitis Polioviruses Polymerase Property Proteins Publishing RNA RNA chemical synthesis Reaction Research Research Personnel Resistance Role Route Secure Series Solubility Solutions Structure Structure-Activity Relationship Syndrome Testing Time Toxic effect Universities Vaccines Viral Virus Virus Replication Work alternative treatment anthrax edema factor base design effective therapy experience gastrointestinal system gel electrophoresis high reward high risk high throughput screening inhibitor/antagonist member mutant nanoparticle nervous system disorder novel nucleotide metabolism pharmacophore poliovirus polymerase 3Dpol polymerization pressure prevent public health relevance research study screening small molecule success uptake

项目摘要

DESCRIPTION (provided by applicant): Persistent infections with Human enteroviruses (HEV) are a major cause of dilated cardiomyopathies, and have been causally implicated in type 1 diabetes, gastrointestinal and nervous system disorders. HEVs are acid-resistant, and can survive in the gastrointestinal tract and other organs for long periods of time. There are currentl no drugs approved to treat HEV infections. The goal of this high-risk/high-reward project is to identify inhibitors of uridylylation of a small protein, VPg, and show that these compounds provide an alternative treatment route for treating chronic infections with slow replicating HEV strains. As uridylylation is virus-specific, the inhibitors should have fewer side effects on mammalian cells. This translational work will combine computational, structural and high throughput screening (HTS) methods to select inhibitors that interfere with uridylylation, and HEV replication. Uridylylation inhibitors selected, in the R21 phase, will be tested, in the R33 phase, for their effects on inhibiting HEV in mammalian cells and in a murine model of chronic HEV infection. Aim 1 is to develop uridylylation assays using a consensus VPg, and screen a diversity set of compounds for those that interfere with this reaction. Active compounds identified in the HTS will be characterized in Aim 2 to derive structure-activity relationships and identify substructures for screening large, web-based compound libraries. A pharmacophore based on VPg structures will be used for further in silico screening of these libraries. Selected compounds will be purchased and assayed, or produced in-house. Assuming that 3-10 uridylylation inhibitors have been identified, the R33 phase of the work will begin. Aim 3 is to further characterize the activity of the uridylylation inhibitors and select those most suitable fo biological assays according to their pharmaceutical properties (low toxicity, good solubility and high stability in cell culture media). Compounds may be formulated in nanoparticles to enhance cell uptake. The most potent, biologically acceptable inhibitors will be further tested for their ability to diminish the replication of HEV strains in infected cells, alone or in combination with other active compounds and antiviral agents. Solution studies using NMR and native gel electrophoresis, and co-crystal structures of the polymerase with the best antiviral compounds will be used to establish their mode of activity. Finally, the antivirals will be tested for their bility to halt HEV replication in cultured cardiomyocytes and a murine model of chronic infection with Coxsackie virus B3. Successful inhibitors will be potential new therapies for HEV infections, that can be used alone or in combination with other compounds developed as part of the Global Polio Eradication Initiative, including capsid binding compounds that inhibit cell entry. Anti-HEV compounds that target uridylylation may thus also play a role in securing polio eradication.

描述（由申请方提供）：人肠道病毒（HEV）持续感染是扩张型心肌病的主要原因，并与1型糖尿病、胃肠道和神经系统疾病有因果关系。HEV是耐酸的，可以在胃肠道和其他器官中存活很长一段时间。目前没有药物被批准用于治疗HEV感染。这个高风险/高回报项目的目标是鉴定小蛋白VPg尿苷酰化的抑制剂，并表明这些化合物为治疗慢性感染慢复制型HEV毒株提供了替代治疗途径。由于尿苷酰化是病毒特异性的，因此抑制剂对哺乳动物细胞的副作用较少。这项翻译工作将结合联合收割机计算，结构和高通量筛选（HTS）的方法来选择抑制剂，干扰尿苷酰化，和HEV复制。在R21阶段选择的尿苷酰化抑制剂将在R33阶段测试其在哺乳动物细胞和慢性HEV感染的鼠模型中抑制HEV的效果。目的1是开发使用共有VPg的尿苷酰化测定，并筛选干扰该反应的化合物的多样性集合。在HTS中鉴定的活性化合物将在目标2中表征，以得出结构-活性关系，鉴定用于筛选大型基于网络的化合物库的子结构。基于VPg结构的药效团将用于这些文库的进一步计算机筛选。所选化合物将购买并进行分析，或在内部生产。假设已经鉴定出3-10种尿苷酰化抑制剂，则将开始R33阶段的工作。目的3是进一步表征尿苷酰化抑制剂的活性，并根据其药物特性（低毒性、良好的溶解性和在细胞培养基中的高稳定性）选择最适合用于生物测定的那些。化合物可以配制成纳米颗粒以增强细胞摄取。将进一步测试最有效的生物学上可接受的抑制剂单独或与其他活性化合物和抗病毒剂组合减少感染细胞中HEV毒株复制的能力。使用NMR和天然凝胶电泳的溶液研究以及聚合酶与最佳抗病毒化合物的共晶体结构将用于建立其活性模式。最后，将测试抗病毒药物在培养的心肌细胞和科萨基病毒B3慢性感染的小鼠模型中阻止HEV复制的能力。成功的抑制剂将是HEV感染的潜在新疗法，可以单独使用或与作为全球根除脊髓灰质炎倡议的一部分开发的其他化合物联合使用，包括抑制细胞进入的衣壳结合化合物。因此，靶向尿苷酰化的抗HEV化合物也可能在确保根除脊髓灰质炎方面发挥作用。