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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：
10318299
负责人：
HONGMIN LI
金额：
$ 56.34万
依托单位：
UNIVERSITY OF ARIZONA
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-02-15 至 2023-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10318299
关键词：
Acquired Immunodeficiency Syndrome Active Sites Address Affect Affinity Animals Antiviral Agents Antiviral Therapy Arboviruses Area Binding Biological Assay Biological Availability Biological Models Cell Culture Techniques Cells Charge Cleaved cell Clinical Complement Complex Dengue Dengue Hemorrhagic Fever Dengue Virus Development Development Plans Disease Disease Outbreaks Drug Kinetics Drug Targeting Ensure Enzyme Inhibitor Drugs Enzyme Interaction Enzymes Flavivirus Flavivirus Infections Fluorescence Resonance Energy Transfer Generations Genes Goals Growth Hepatitis C Infection Japanese Encephalitis Vaccines Japanese encephalitis virus Lead Libraries Life Luciferases Mass Vaccinations Meningitis Microcephaly Modeling Multienzyme Complexes Mus Mutagenesis Mutation Outcome Study Peptide Hydrolases Pharmaceutical Preparations Polyproteins Populations at Risk Prevention Protease Inhibitor Proteins Public Health Renilla Luciferases Replicon Research Resistance Resources Site Solid Structure Structure-Activity Relationship System Technology Testing Therapeutic Tick-Borne Encephalitis Virus Toxic effect Toxicity Tests 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 Zika Virus anti-viral efficacy arthropod-borne base burden of illness combat drug development drug discovery efficacy evaluation fighting high throughput screening human disease human pathogen in vivo inhibitor/antagonist innovation nanomolar novel public health priorities screening small molecule libraries therapeutic target therapy design tool vaccine development viral resistance virology

项目摘要

Project Summary/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. The long- range goal of this proposal is to address this need by developing a new system to identify, screen and validate a class of potential flavivirus inhibitors, and successfully identify and analyze candidate compounds that show promise in disrupting viral survival. During flaviviral 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 objectives of this project are to develop innovative high throughput screening strategies to identify and characterize compounds that orthosterically inhibit the function of the critical protease NS2B- NS3. To achieve project objectives, in Specific Aim 1, we will develop and perform novel high- throughput screening primary, secondary, and tertiary assays capable of screening large chemical libraries to identify orthosteric inhibitors. In Specific Aim 2, after optimizing these assays, we will perform large-scale HTS to screen a compound library of >400,000 compounds for potential protease inhibitor candidates, employing five levels of screening to fully eliminate false positives. In Specific Aim 3, we will evaluate the efficacy of the resulting candidate compounds for their capacity to block protease activity, and determined their modes of actions. Lead compounds against the protease will be tested for cellular toxicity, reduction of flavivirus titer in cell culture, resistant variants, and limited structure-activity relationship. We will ultimately test the toxicity, pharmacokinetics, and efficacy in live animals for the most potent compounds. We anticipate the outcome of this study will be the generation of crucial new information on flavivirus inhibitor-enzyme interaction at structurally significant sites. This creates the potential for identifying novel classes of flaviviral inhibitors, suitable for addressing the global public health need for new antiviral agents to fight flaviviruses.

项目总结/文摘: