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

• 批准号: 10376239
• 负责人: HONGMIN LI
• 金额: $ 55万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-02-15 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10376239
• 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; Clinical; Complement; Complex; Dengue; Dengue Hemorrhagic Fever; Dengue Virus; 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; Persons; Pharmaceutical Preparations; Polyproteins; Populations at Risk; Prevention; Protease Inhibitor; Proteins; Public Health; Renilla Luciferases; Replicon; Research; Resistance; Resources; Site; Solid; Structure-Activity Relationship; System; Technology; Testing; 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; antiviral drug development; arthropod-borne; base; burden of illness; combat; drug development; drug discovery; efficacy evaluation; fighting; high throughput screening; human disease; human pathogen; in vivo; inhibitor; innovation; nanomolar; novel; public health priorities; screening; small molecule libraries; therapeutic target; therapy design; tool; vaccine development; viral resistance; virology

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:

项目成果

Calcimycin Inhibits Cryptococcus neoformans In Vitro and In Vivo by Targeting the Prp8 Intein Splicing.

• DOI: 10.1021/acsinfecdis.2c00137
• 发表时间: 2022-09-09
• 期刊: ACS INFECTIOUS DISEASES
• 影响因子: 5.3
• 作者: Tharappel, Anil Mathew;Li, Zhong;Zhu, Yan Chun;Wu, Xiangmeng;Chaturvedi, Sudha;Zhang, Qing-Yu;Li, Hongmin
• 通讯作者: Li, Hongmin

Erythrosin B is a potent and broad-spectrum orthosteric inhibitor of the flavivirus NS2B-NS3 protease.

• DOI: 10.1016/j.antiviral.2017.12.018
• 发表时间: 2018-03
• 期刊: Antiviral research
• 影响因子: 7.6
• 作者: Li Z;Sakamuru S;Huang R;Brecher M;Koetzner CA;Zhang J;Chen H;Qin CF;Zhang QY;Zhou J;Kramer LD;Xia M;Li H
• 通讯作者: Li H

Methylene blue is a potent and broad-spectrum inhibitor against Zika virus in vitro and in vivo.

• DOI: 10.1080/22221751.2020.1838954
• 发表时间: 2020-12
• 期刊: Emerging microbes & infections
• 影响因子: 13.2
• 作者: Li Z;Lang Y;Sakamuru S;Samrat S;Trudeau N;Kuo L;Rugenstein N;Tharappel A;D'Brant L;Koetzner CA;Hu S;Zhang J;Huang R;Kramer LD;Butler D;Xia M;Li H
• 通讯作者: Li H

In vitro and in vivo characterization of erythrosin B and derivatives against Zika virus.

• DOI: 10.1016/j.apsb.2021.10.017
• 发表时间: 2022-04
• 期刊: ACTA PHARMACEUTICA SINICA B
• 影响因子: 14.5
• 作者: Li, Zhong;Xu, Jimin;Lang, Yuekun;Wu, Xiangmeng;Hu, Saiyang;Samrat, Subodh Kumar;Tharappel, Anil M.;Kuo, Lili;Butler, David;Song, Yongcheng;Zhang, Qing-Yu;Zhou, Jia;Li, Hongmin
• 通讯作者: Li, Hongmin

JMX0207, a Niclosamide Derivative with Improved Pharmacokinetics, Suppresses Zika Virus Infection Both In Vitro and In Vivo.

• DOI: 10.1021/acsinfecdis.0c00217
• 发表时间: 2020-10-09
• 期刊: ACS infectious diseases
• 影响因子: 5.3
• 作者: Li Z;Xu J;Lang Y;Fan X;Kuo L;D'Brant L;Hu S;Samrat SK;Trudeau N;Tharappel AM;Rugenstein N;Koetzner CA;Zhang J;Chen H;Kramer LD;Butler D;Zhang QY;Zhou J;Li H
• 通讯作者: Li H