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：
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
项目类别：
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

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.

项目摘要/摘要：许多黄病毒，如登革热病毒、寨卡病毒、西尼罗河病毒和黄热病病毒导致严重的人类疾病。然而，尚无临床批准的抗病毒疗法用于治疗黄病毒感染。因此，疫苗和抗病毒药物的开发预防和治疗黄病毒感染是一个明确的公共卫生优先事项。长- 该提案的目标是通过开发一个新系统来识别、满足这一需求，筛选并验证一类潜在的黄病毒抑制剂，并成功识别和分析有望破坏病毒存活的候选化合物。在黄病毒感染宿主细胞期间，由病毒合成单个病毒多蛋白基因组。这种多蛋白通过宿主细胞的组合被切割成其组成蛋白蛋白酶和双组分病毒蛋白酶，编码在基因 NS2B 和 NS3 中。功能这种病毒蛋白酶对于病毒组装和复制是必不可少的。本次活动的目标项目将开发创新的高通量筛选策略来识别和表征正位抑制关键蛋白酶 NS2B- 功能的化合物 NS3。为了实现项目目标，在具体目标 1 中，我们将开发并执行新颖的高通量筛选初级、二级和三级测定能够筛选大量化学库来识别正位抑制剂。在具体目标2中，优化这些之后分析中，我们将进行大规模 HTS 来筛选超过 400,000 种化合物的化合物库对于潜在的蛋白酶抑制剂候选者，采用五级筛选以完全消除误报。在具体目标 3 中，我们将评估最终候选人的功效化合物阻断蛋白酶活性的能力，并确定其作用方式。将测试针对蛋白酶的先导化合物的细胞毒性和黄病毒的减少细胞培养物中的滴度、耐药变异以及有限的构效关系。我们最终将在活体动物中测试最有效化合物的毒性、药代动力学和功效。我们预计这项研究的结果将是产生关于以下方面的重要新信息：黄病毒抑制剂-酶在结构重要位点的相互作用。这创造了潜力用于识别适合全球公众的新型黄病毒抑制剂健康需要新的抗病毒药物来对抗黄病毒。

项目成果

期刊论文数量（12）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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

赤藓红 B 及其衍生物抗寨卡病毒的体外和体内表征。

DOI：
发表时间：
2022-04
期刊：
影响因子：
0
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;Zhou, Jia;Li, Hongmin
通讯作者：
Li, Hongmin

Broad-Spectrum Small-Molecule Inhibitors Targeting the SAM-Binding Site of Flavivirus NS5 Methyltransferase.

针对黄病毒 NS5 甲基转移酶 SAM 结合位点的广谱小分子抑制剂。

DOI：
10.1021/acsinfecdis.2c00571
发表时间：
2023-06-22
期刊：
ACS infectious diseases
影响因子：
5.3
作者：
S. Samrat;Q. Bashir;Yiding Huang;Carl William Trieshmann;Anil M. Tharappel;Ran Zhang;Ke Chen;Y. G. Zheng;Zhong Li;Hongmin Li
通讯作者：
Hongmin Li

Analysis of Protein-Protein Interactions by Split Luciferase Complementation Assay.

通过裂解荧光素酶互补测定分析蛋白质-蛋白质相互作用。