权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Antiviral potential of helicase inhibitors

解旋酶抑制剂的抗病毒潜力

基本信息

批准号：
8211062
负责人：
David N Frick
金额：
$ 36.56万
依托单位：
UNIVERSITY OF WISCONSIN MILWAUKEE
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-02-01 至 2014-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8211062
关键词：
ATP Hydrolysis ATP phosphohydrolase Acquired Immunodeficiency Syndrome Antiviral Agents Basic Science Biological Assay Boxing Categories Cells Chemicals Chemistry Clinic Clinical Clinical Trials Collaborations Collection DDX1 gene DDX1 protein DNA DNA Binding DNA Viruses Data Dengue Dengue Virus Development Direct Costs Drug Delivery Systems Enzymes Flavivirus Funding Funding Opportunities Future Goals HIV Hepatitis C Hepatitis C virus Human Japanese Encephalitis Japanese Encephalitis Viruses Japanese encephalitis virus Kansas Learning Ligand Binding Link Molecular Molecular Bank Monitor New York Patients Peptide Hydrolases Peptides Pharmaceutical Preparations Polymerase Preclinical Drug Evaluation Production Proteins RNA RNA Viruses RNA helicase A RNA replication Research Institute Research Project Grants Role Screening procedure Specificity Testing Translating United States National Institutes of Health Virus Virus Replication West Nile virus Yellow Fever Yellow fever virus anti-hepatitis C base biodefense burden of illness design drug development helicase high throughput screening human WRN protein inhibitor/antagonist innovation medical schools pathogen public health relevance response scale up small molecule tool viral RNA

项目摘要

DESCRIPTION (provided by applicant): In response to the Funding Opportunity Announcement "Development of Assays for High-Throughput Drug Screening," we propose here to develop assays to identify and analyze inhibitors of helicases needed for RNA virus replication. All viruses require helicases to separate duplex DNA or RNA, yet helicases remain undeveloped antiviral drug targets. Although some helicase inhibitors that target DNA viruses are progressing in clinical trials, no antiviral agents targeting RNA viruses have entered the clinical arena. We therefore will target RNA viruses, focusing on pathogens with abnormally high global burdens of disease: hepatitis C virus (HCV), Dengue virus (DENV), West Nile virus (WNV), Japanese encephalitis virus (JEV), yellow fever virus (YFV), and human immunodeficiency virus (HIV). The assays to be developed are needed as part of an ongoing collaborative hypothesis-driven research project to develop HCV NS3 helicase inhibitors as chemical probes to facilitate basic science and for drug development. In that endeavor, we have initiated an uHTS at the MLPCN that uses our recently developed molecular beacon based helicase assay (MBHA) as a primary screen. To evaluate HCV helicase inhibitor specificity, MBHAs will be developed using helicases encoded by viruses related to HCV (DNV, WNV, YFV, and JEV) and human cellular helicases. The human proteins to be analyzed have all been linked to HIV replication and include the DEAD-box proteins DDX1, DDX3, DDX24, MDA-5, RNA helicase A (RHA), and Werner syndrome protein (WRN). In aim 2, assays will be developed to analyze the cellular antiviral efficacy, and mechanism of action of HCV helicase inhibitors. Mechanistic assays include ATPase assays, ligand binding assays, protein oligomerization assays, and simultaneous protease/helicase assays. Most of these mechanistic assays could be used with either HCV helicase or one of the ten other helicases targeted here to screen for additional classes of helicase inhibitors. PUBLIC HEALTH RELEVANCE: In collaboration with a NIH high throughput-screening center, my lab is searching a collection of hundreds of thousands of small molecules for compounds that might inhibit the helicase encoded by the hepatitis C virus (HCV). In this project, we will develop tests to rapidly search these HCV helicase inhibitors for compounds that could be used to treat hepatitis C. We will also develop similar assays to identify compounds that might inhibit viruses that cause AIDS, yellow fever, Dengue fever, West Nile and Japanese encephalitis.

描述（由申请人提供）：为了响应资助机会公告“开发高通量药物筛选测定法”，我们在此建议开发测定法来识别和分析 RNA 病毒复制所需的解旋酶抑制剂。所有病毒都需要解旋酶来分离双链 DNA 或 RNA，但解旋酶仍然是尚未开发的抗病毒药物靶标。尽管一些针对DNA病毒的解旋酶抑制剂正在临床试验中取得进展，但还没有针对RNA病毒的抗病毒药物进入临床舞台。因此，我们将针对RNA病毒，重点关注全球疾病负担异常高的病原体：丙型肝炎病毒（HCV）、登革热病毒（DENV）、西尼罗河病毒（WNV）、日本脑炎病毒（JEV）、黄热病病毒（YFV）和人类免疫缺陷病毒（HIV）。待开发的检测方法是正在进行的假设驱动合作研究项目的一部分，该项目旨在开发 HCV NS3 解旋酶抑制剂作为化学探针，以促进基础科学和药物开发。在这一努力中，我们在 MLPCN 启动了 uHTS，使用我们最近开发的基于分子信标的解旋酶测定 (MBHA) 作为主要筛选。为了评估 HCV 解旋酶抑制剂的特异性，将使用 HCV 相关病毒（DNV、WNV、YFV 和 JEV）和人类细胞解旋酶编码的解旋酶来开发 MBHA。待分析的人类蛋白均与 HIV 复制有关，包括 DEAD-box 蛋白 DDX1、DDX3、DDX24、MDA-5、RNA 解旋酶 A (RHA) 和维尔纳综合征蛋白 (WRN)。在目标 2 中，将开发分析 HCV 解旋酶抑制剂的细胞抗病毒功效和作用机制的测定方法。机制测定包括 ATP 酶测定、配体结合测定、蛋白质寡聚化测定和同步蛋白酶/解旋酶测定。大多数这些机制测定可与 HCV 解旋酶或此处针对的十种其他解旋酶之一一起使用，以筛选其他类别的解旋酶抑制剂。公共健康相关性：我的实验室与 NIH 高通量筛选中心合作，正在数十万种小分子中寻找可能抑制丙型肝炎病毒 (HCV) 编码的解旋酶的化合物。在这个项目中，我们将开发测试来快速搜索这些HCV解旋酶抑制剂，寻找可用于治疗丙型肝炎的化合物。我们还将开发类似的分析方法来识别可能抑制导致艾滋病、黄热病、登革热、西尼罗河和日本脑炎的病毒的化合物。

项目成果

期刊论文数量（13）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（1）

Role of the Conserved DECH-Box Cysteine in Coupling Hepatitis C Virus Helicase-Catalyzed ATP Hydrolysis to RNA Unwinding.

保守的 DECH-Box 半胱氨酸在丙型肝炎病毒解旋酶催化 ATP 水解与 RNA 解旋偶联中的作用。

DOI：
10.1021/acs.biochem.8b00796
发表时间：
2018
期刊：
Biochemistry
影响因子：
2.9
作者：
Yerukhimovich,MarkM;Marohnic,ChristopherC;Frick,DavidN
通讯作者：
Frick,DavidN

Discovering new medicines targeting helicases: challenges and recent progress.

发现靶向解旋酶的新药物：挑战和最新进展。

DOI：
10.1177/1087057113482586
发表时间：
2013-08
期刊：
Journal of biomolecular screening
影响因子：
0
作者：
Shadrick WR;Ndjomou J;Kolli R;Mukherjee S;Hanson AM;Frick DN
通讯作者：
Frick DN

New Techniques to Study Intracellular Receptors in Living Cells: Insights Into RIG-I-Like Receptor Signaling.

研究活细胞内受体的新技术：深入了解 RIG-I 样受体信号传导。

DOI：
10.1007/5584_2018_297
发表时间：
2019
期刊：
Advances in experimental medicine and biology
影响因子：
0
作者：
Corby,MJ;Raicu,Valerica;Frick,DavidN
通讯作者：
Frick,DavidN

Discovery of the 2-phenyl-4,5,6,7-Tetrahydro-1H-indole as a novel anti-hepatitis C virus targeting scaffold.

DOI：
10.1016/j.ejmech.2015.04.022
发表时间：
2015
期刊：
European journal of medicinal chemistry
影响因子：
6.7
作者：
Andreev IA;Manvar D;Barreca ML;Belov DS;Basu A;Sweeney NL;Ratmanova NK;Lukyanenko ER;Manfroni G;Cecchetti V;Frick DN;Altieri A;Kaushik-Basu N;Kurkin AV
通讯作者：
Kurkin AV

Analysis of the Enzymatic Activity of an NS3 Helicase Genotype 3a Variant Sequence Obtained from a Relapse Patient.