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Molecular Modulators of HCV Replication

HCV 复制的分子调节剂

基本信息

批准号：
7169837
负责人：
Neerja Kaushik-Basu
金额：
$ 25.8万
依托单位：
UNIV OF MED/DENT OF NJ-NJ MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-04-01 至 2010-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7169837
关键词：
Amino Acids Binding Sites Biochemical Biological Assay C-terminal Cell Culture System Cell Line Cells Characteristics Complex Consensus Sequence Conserved Sequence Cultured Cells Development Drug Design Elements Enzymatic Biochemistry Enzymes Event Family Genetic Transcription Genome Genomics Glutathione S-Transferase Goals Hepatitis C virus Human Human Cell Line In Vitro Kinetics Knowledge Lead Liver Luciferases Mediating Membrane Molecular Mutation Nonstructural Protein Nucleosides Numbers Open Reading Frames Pharmaceutical Preparations Phenylalanine Play Polymerase Process Production Protein Microchips Proteins Proteomics RNA RNA Viruses RNA chemical synthesis RNA replication RNA-Directed RNA Polymerase Range Regulation Replication-Associated Process Replicon Reporter Ribonucleoproteins Roentgen Rays Role Screening procedure Series Site-Directed Mutagenesis Source Structural Protein Structure Structure-Activity Relationship Surface Plasmon Resonance System Tail Technology Tissues Viral Viral Genome Viral Proteins Virion Virus Virus Diseases Virus Replication abstracting base cDNA Library combinatorial design enzyme mechanism in vivo inhibitor/antagonist insight mutant new technology novel novel strategies nucleoside inhibitor programs protein protein interaction replicase three dimensional structure transmission process viral RNA virus host interaction virus tropism yeast two hybrid system

项目摘要

(Revised Abstract) DESCRIPTION: The molecular events that mediate replication of hepatitis C virus (HCV) are largely obscure. The HCV nonstructural protein, NS5B, that synthesizes the viral positive strand RNA genome via (-) strand intermediate, is believed to be the key enzyme responsible for its replication but there are many unanswered questions about how its activity is controlled. Moreover, very few host cell protein(s)/factor(s) and other non-structural (NS) HCV proteins, which interact with NS5B and modulate its function, synthesis or turnover, have been identified and characterized. Proposed here, therefore, are two broad aspects, structure-activity relationship (SAR) optimization studies to design, and synthesize novel NS5B specific inhibitors and identification of cellular/viral protein(s) facilitating NS5B's activity upon interaction. In Aim I, we will explore the potential of phenylalanine-derived non-nucleoside compounds in inhibiting NS5B's enzymatic activity. These studies will be guided by the resolved crystal structures of NS5B and its similarities with other polymerase's structures. NS5B contains a hydrophobic C-terminal tail that aids in membrane integration of the protein to form a replisome complex which is associated with cytoskeletal elements. However, the physical and functional interactions between NS5B and the cellular and other viral components of this complex are poorly understood. In Aim II we propose to identify HCV nonstructural proteins interacting with NS5B using the ciphergen proteomic technology. For this, we will employ the UHCV-11 cell line, inducibly expressing the entire HCV open reading frame except NS5B. The kinetic parameters governing these interactions will be validated via surface plasmon resonance (Biacore). In Aim III, we propose to identify host cell protein(s) from normal and HCV-infected liver explants, interacting with NS5B and thereby modulating its catalytic function. A critical understanding of NS5B, in perspective of its structure-function relationships and its combinatorial interaction with other proteins, will provide insights into the molecular effectors mediating HCV replication. This will enable us to understand, at the molecular level, the replication process of hepatitis C virus and will further facilitate the development of effective drugs/inhibitors against HCV.

（修订摘要）描述：介导丙型肝炎病毒（HCV）复制的分子事件在很大程度上是模糊的。HCV非结构蛋白NS 5 B通过（-）链中间体合成病毒正链RNA基因组，被认为是负责其复制的关键酶，但关于其活性如何控制还有许多未解的问题。此外，已经鉴定和表征了非常少的宿主细胞蛋白/因子和其他非结构（NS）HCV蛋白，其与NS 5 B相互作用并调节其功能、合成或周转。因此，本文提出了两个广泛的方面，设计和合成新型NS 5 B特异性抑制剂的结构-活性关系（SAR）优化研究以及促进NS 5 B在相互作用时的活性的细胞/病毒蛋白的鉴定。在目的I中，我们将探索苯丙氨酸衍生的非核苷化合物在抑制NS 5 B的酶活性方面的潜力。这些研究将由NS 5 B的解析晶体结构及其与其他聚合酶结构的相似性指导。NS 5 B含有疏水性C末端尾，其有助于蛋白质的膜整合以形成与细胞骨架元件相关的复制体复合物。然而，NS 5 B与该复合物的细胞和其他病毒组分之间的物理和功能相互作用知之甚少。在目的II中，我们建议使用密码子蛋白质组学技术鉴定与NS 5 B相互作用的HCV非结构蛋白。为此，我们将使用UHCV-11细胞系，其可诱导表达除NS 5 B之外的整个HCV开放阅读框架。控制这些相互作用的动力学参数将通过表面等离子体共振（Biacore）进行验证。在目的III中，我们提出从正常和HCV感染的肝外植体中鉴定宿主细胞蛋白，其与NS 5 B相互作用，从而调节其催化功能。从NS 5 B的结构-功能关系及其与其他蛋白质的组合相互作用的角度，对NS 5 B的关键性理解将为介导HCV复制的分子效应器提供深入了解。这将使我们能够在分子水平上了解丙型肝炎病毒的复制过程，并将进一步促进开发有效的抗HCV药物/抑制剂。

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Structure-based virtual screening, synthesis and SAR of novel inhibitors of hepatitis C virus NS5B polymerase.

DOI：
10.1016/j.bmc.2010.05.030
发表时间：
2010-07-01
期刊：
BIOORGANIC & MEDICINAL CHEMISTRY
影响因子：
3.5
作者：
Talele, Tanaji T.;Arora, Payal;Kulkarni, Shridhar S.;Patel, Maulik R.;Singh, Satyakam;Chudayeu, Maksim;Kaushik-Basu, Neerja
通讯作者：
Kaushik-Basu, Neerja

Identification and characterization of coumestans as novel HCV NS5B polymerase inhibitors.

鉴定和表征香豆作为新型HCV NS5B聚合酶抑制剂。

DOI：
10.1093/nar/gkm1178
发表时间：
2008-03
期刊：
NUCLEIC ACIDS RESEARCH
影响因子：
14.9
作者：
Kaushik-Basu, Neerja;Bopda-Waffo, Alain;Talele, Tanaji T.;Basu, Amartya;Costa, Paulo R. R.;da Silva, Alcides J. M.;Sarafianos, Stefan G.;Noel, Francois
通讯作者：
Noel, Francois

Non-nucleoside inhibitors of the hepatitis C virus NS5B RNA-dependant RNA polymerase: 2-aryl-3-heteroaryl-1,3-thiazolidin-4-one derivatives.

丙型肝炎病毒 NS5B RNA 依赖性 RNA 聚合酶的非核苷抑制剂：2-芳基-3-杂芳基-1,3-噻唑烷-4-酮衍生物。

DOI：
10.1016/j.bmcl.2008.10.023
发表时间：
2008
期刊：
Bioorganic & medicinal chemistry letters
影响因子：
2.7
作者：
Rawal,RavindraK;Katti,SB;Kaushik-Basu,Neerja;Arora,Payal;Pan,Zhenhua
通讯作者：
Pan,Zhenhua

Etodolac Thiosemicarbazides: A novel class of hepatitis C virus NS5B polymerase inhibitors

DOI：
10.12991/201317382
发表时间：
2013-01-01
期刊：
MARMARA PHARMACEUTICAL JOURNAL
影响因子：
0
作者：
Cikla, Pelin;Arora, Payal;Kucukguzel, S. Guniz
通讯作者：
Kucukguzel, S. Guniz

Persistent growth of a human plasma-derived hepatitis C virus genotype 1b isolate in cell culture.

人血浆来源的丙型肝炎病毒基因型 1b 分离株在细胞培养物中持续生长。

DOI：
10.1371/journal.ppat.1000910
发表时间：
2010
期刊：
PLoS pathogens
影响因子：
6.7
作者：
Silberstein,Erica;Mihalik,Kathleen;Ulitzky,Laura;Plant,EwanP;Puig,Montserrat;Gagneten,Sara;Yu,Mei-yingW;Kaushik-Basu,Neerja;Feinstone,StephenM;Taylor,DeborahR
通讯作者：
Taylor,DeborahR