High throughput screening of orthosteric inhibitors of flavivirus protease

黄病毒蛋白酶正构抑制剂的高通量筛选

基本信息

批准号：
9538444
负责人：
HONGMIN LI
金额：
$ 52.36万
依托单位：
WADSWORTH CENTER
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-08-18 至 2018-02-14
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9538444
关键词：
Acquired Immunodeficiency Syndrome Active Sites Affect Affinity Animals Antiviral Agents Antiviral Therapy Arboviruses Area Arthropods Binding Biological Assay Biological Availability Biological Models Cell Culture Techniques Cells Charge Cleaved cell Clinical Complement Complex Dengue Dengue Hemorrhagic Fever Dengue Virus Development Disease Disease Outbreaks Drug Targeting Encephalitis Viruses Ensure Enzyme Inhibitor Drugs Enzyme Interaction Enzymes Flavivirus Flavivirus Infections Fluorescence Genes Goals Growth Hepatitis C Human Infection Japanese Encephalitis Vaccines Japanese encephalitis virus Lead Libraries Life Luciferases Mass Vaccinations Meningitis Microcephaly Modeling Molecular Conformation Mutagenesis Mutation Peptide Hydrolases Performance Phase Polyproteins Populations at Risk Prevention Protease Inhibitor Proteins Research Resistance Site Solid Technology Testing Therapeutic Tick-Borne Encephalitis Virus Tick-Borne Encephalitis Viruses Toxic effect Vaccinated Vaccines Variant Viral Viral Genome Viral Physiology Viral Proteins Virus Virus Assembly Virus Replication West Nile Encephalitis West Nile virus Work Yellow fever virus Zika Virus base burden of illness combat drug development drug discovery experience high throughput screening human disease in vivo inhibitor/antagonist innovation killings nanomolar novel pathogen public health priorities screening small molecule libraries therapeutic target tool vaccine development viral resistance virology

项目摘要

Abstract: Many flaviviruses such as Dengue virus, Zika virus, West Nile virus, and Yellow Fever virus cause significant human diseases. However, no clinically approved antiviral therapy is available for treatment of flavivirus infections. Therefore, the development of vaccines and antiviral agents for prevention and treatment of flavivirus infections is a clear public health priority. During infection of the host cell, a single viral polyprotein is synthesized from the viral genome. This polyprotein is cleaved into its component proteins by a combination of host cell proteases and a two-component viral protease, encoded in genes NS2B and NS3. The function of this viral protease is indispensable for virus assembly and replication. The goal of this proposal is to develop high throughput screening strategies to identify and characterize compounds that orthosterically inhibit the function of the critical protease. In Specific Aims 1 and 2 we will develop and perform novel high-throughput screening assays capable of screening large chemical libraries to identify orthosteric inhibitors. In Specific Aim 3 we will evaluate the efficacy of the resulting candidate compounds for their capacity to block protease activity. Lead compounds against the protease will be tested for cellular toxicity, reduction of flavivirus titer in cell culture, and resistant variants. We will ultimately test the best lead compound toxicity and efficacy in live animals for the most potent compounds. This study will generate information on flavivirus inhibitor-enzyme interaction at structurally significant sites and potentially lead to novel classes of flaviviral inhibitors.

摘要：许多黄素病毒，例如登革热病毒，寨卡病毒，西尼罗河病毒和黄热病病毒重大的人类疾病。但是，尚无临床批准的抗病毒疗法可用于治疗黄病毒感染。因此，开发预防疫苗和抗病毒剂黄病毒感染的治疗是明显的公共卫生优先事项。在宿主细胞感染期间，一个病毒多蛋白是从病毒基因组合成的。该多蛋白被裂解成其成分蛋白质通过宿主细胞蛋白酶和两种组分病毒蛋白酶的组合，在基因中编码 NS2B和NS3。这种病毒蛋白酶的功能对于病毒组装和复制是必不可少的。这该建议的目标是制定高吞吐量筛选策略以识别和表征正向抑制关键蛋白酶功能的化合物。在特定目标1和2中我们将开发和执行能够筛选大化学物质的新型高通量筛选测定法识别正态抑制剂的库。在特定目标3中，我们将评估结果的功效候选化合物具有阻断蛋白酶活性的能力。针对蛋白酶的铅化合物将测试细胞毒性，细胞培养中黄病毒滴度的降低以及耐药性变体。我们将最终测试活动物中最佳的铅复合毒性和功效化合物。这项研究将在结构上生成有关黄病毒抑制剂 - 酶相互作用的信息重要的部位并有可能导致新型的黄病毒抑制剂。