High-throughput assay using a GFP-expressing replicon for SARS-CoV drug discovery.
Identifieur interne : 001B13 ( PubMed/Corpus ); précédent : 001B12; suivant : 001B14High-throughput assay using a GFP-expressing replicon for SARS-CoV drug discovery.
Auteurs : Feng Ge ; Sheng Xiong ; Fu-Sen Lin ; Zhi-Ping Zhang ; Xian-En ZhangSource :
- Antiviral research [ 1872-9096 ] ; 2008.
English descriptors
- KwdEn :
- Animals, Antiviral Agents (pharmacology), Cell Line, Chlorocebus aethiops, Cricetinae, Drug Evaluation, Preclinical, Green Fluorescent Proteins (genetics), Green Fluorescent Proteins (metabolism), Humans, Replicon, SARS Virus (drug effects), SARS Virus (genetics), Severe Acute Respiratory Syndrome (drug therapy), Severe Acute Respiratory Syndrome (virology), Small Molecule Libraries (pharmacology), Vero Cells, Virus Replication (drug effects).
- MESH :
- chemical , genetics : Green Fluorescent Proteins.
- chemical , metabolism : Green Fluorescent Proteins.
- chemical , pharmacology : Antiviral Agents, Small Molecule Libraries.
- drug effects : SARS Virus, Virus Replication.
- drug therapy : Severe Acute Respiratory Syndrome.
- genetics : SARS Virus.
- virology : Severe Acute Respiratory Syndrome.
- Animals, Cell Line, Chlorocebus aethiops, Cricetinae, Drug Evaluation, Preclinical, Humans, Replicon, Vero Cells.
Abstract
The causative agent of severe acute respiratory syndrome (SARS) has been identified as a novel coronavirus, SARS-CoV. The development of rapid screening assays is essential for antiviral drug discovery. By using a cell line expressing a SARS-CoV subgenomic replicon, we developed a high-throughput assay and used it to screen small molecule compounds for inhibitors of SARS-CoV replication in the absence of live virus. The assay system involves minimal manipulation after assay set-up, facilitates automated read-out and minimizes risks associated with hazardous viruses. Based on this assay system, we screened 7035 small molecule compounds from which we identified 7 compounds with anti-SARS-CoV activity. We demonstrate that the compounds inhibited SARS-CoV replication-dependent GFP expression in the replicon cells and reduced SARS-CoV viral protein accumulation and viral RNA copy number in the replicon cells. In a SARS-CoV plaque reduction assay, these compounds were confirmed to have antiviral activity. The target of one of the hit compounds, C12344, was validated by the generation of resistant replicon cells and the identification of the mutations conferring the resistant phenotype. These compounds should be valuable for developing anti-SARS therapeutic drugs as well as research tools to study the mechanism of SARS-CoV replication.
DOI: 10.1016/j.antiviral.2008.05.005
PubMed: 18584889
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pubmed:18584889Le document en format XML
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The development of rapid screening assays is essential for antiviral drug discovery. By using a cell line expressing a SARS-CoV subgenomic replicon, we developed a high-throughput assay and used it to screen small molecule compounds for inhibitors of SARS-CoV replication in the absence of live virus. The assay system involves minimal manipulation after assay set-up, facilitates automated read-out and minimizes risks associated with hazardous viruses. Based on this assay system, we screened 7035 small molecule compounds from which we identified 7 compounds with anti-SARS-CoV activity. We demonstrate that the compounds inhibited SARS-CoV replication-dependent GFP expression in the replicon cells and reduced SARS-CoV viral protein accumulation and viral RNA copy number in the replicon cells. In a SARS-CoV plaque reduction assay, these compounds were confirmed to have antiviral activity. The target of one of the hit compounds, C12344, was validated by the generation of resistant replicon cells and the identification of the mutations conferring the resistant phenotype. These compounds should be valuable for developing anti-SARS therapeutic drugs as well as research tools to study the mechanism of SARS-CoV replication.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">18584889</PMID><DateCompleted><Year>2008</Year><Month>11</Month><Day>11</Day></DateCompleted><DateRevised><Year>2020</Year><Month>04</Month><Day>03</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1872-9096</ISSN><JournalIssue CitedMedium="Internet"><Volume>80</Volume><Issue>2</Issue><PubDate><Year>2008</Year><Month>Nov</Month></PubDate></JournalIssue><Title>Antiviral research</Title><ISOAbbreviation>Antiviral Res.</ISOAbbreviation></Journal><ArticleTitle>High-throughput assay using a GFP-expressing replicon for SARS-CoV drug discovery.</ArticleTitle><Pagination><MedlinePgn>107-13</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1016/j.antiviral.2008.05.005</ELocationID><Abstract><AbstractText>The causative agent of severe acute respiratory syndrome (SARS) has been identified as a novel coronavirus, SARS-CoV. The development of rapid screening assays is essential for antiviral drug discovery. By using a cell line expressing a SARS-CoV subgenomic replicon, we developed a high-throughput assay and used it to screen small molecule compounds for inhibitors of SARS-CoV replication in the absence of live virus. The assay system involves minimal manipulation after assay set-up, facilitates automated read-out and minimizes risks associated with hazardous viruses. Based on this assay system, we screened 7035 small molecule compounds from which we identified 7 compounds with anti-SARS-CoV activity. We demonstrate that the compounds inhibited SARS-CoV replication-dependent GFP expression in the replicon cells and reduced SARS-CoV viral protein accumulation and viral RNA copy number in the replicon cells. In a SARS-CoV plaque reduction assay, these compounds were confirmed to have antiviral activity. The target of one of the hit compounds, C12344, was validated by the generation of resistant replicon cells and the identification of the mutations conferring the resistant phenotype. These compounds should be valuable for developing anti-SARS therapeutic drugs as well as research tools to study the mechanism of SARS-CoV replication.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Ge</LastName><ForeName>Feng</ForeName><Initials>F</Initials><AffiliationInfo><Affiliation>Institute of Life and Health Engineering, Jinan University, Guangzhou, Guangdong, PR China. tgfeng@jnu.edu.cn</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Xiong</LastName><ForeName>Sheng</ForeName><Initials>S</Initials></Author><Author ValidYN="Y"><LastName>Lin</LastName><ForeName>Fu-Sen</ForeName><Initials>FS</Initials></Author><Author ValidYN="Y"><LastName>Zhang</LastName><ForeName>Zhi-Ping</ForeName><Initials>ZP</Initials></Author><Author ValidYN="Y"><LastName>Zhang</LastName><ForeName>Xian-En</ForeName><Initials>XE</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal 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IdType="pubmed">18584889</ArticleId><ArticleId IdType="pii">S0166-3542(08)00311-2</ArticleId><ArticleId IdType="doi">10.1016/j.antiviral.2008.05.005</ArticleId><ArticleId IdType="pmc">PMC7114325</ArticleId></ArticleIdList><ReferenceList><Reference><Citation>Chem Biol. 2004 Sep;11(9):1293-9</Citation><ArticleIdList><ArticleId IdType="pubmed">15380189</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>J Virol Methods. 2005 Oct;129(1):56-63</Citation><ArticleIdList><ArticleId IdType="pubmed">15961169</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>J Virol. 2003 Sep;77(18):9790-8</Citation><ArticleIdList><ArticleId IdType="pubmed">12941887</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Virology. 2007 Mar 30;360(1):150-8</Citation><ArticleIdList><ArticleId IdType="pubmed">17098272</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Proc Natl Acad Sci U S A. 2003 Oct 28;100(22):12995-3000</Citation><ArticleIdList><ArticleId IdType="pubmed">14569023</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>J Gen Virol. 2003 Sep;84(Pt 9):2305-2315</Citation><ArticleIdList><ArticleId IdType="pubmed">12917450</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>FEBS Lett. 2004 Oct 22;576(3):325-30</Citation><ArticleIdList><ArticleId IdType="pubmed">15498556</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>N Engl J Med. 2003 Dec 18;349(25):2431-41</Citation><ArticleIdList><ArticleId IdType="pubmed">14681510</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Science. 2003 May 30;300(5624):1399-404</Citation><ArticleIdList><ArticleId IdType="pubmed">12730501</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>N Engl J Med. 2003 May 15;348(20):1953-66</Citation><ArticleIdList><ArticleId IdType="pubmed">12690092</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>J Virol. 2001 Feb;75(3):1252-64</Citation><ArticleIdList><ArticleId IdType="pubmed">11152498</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Trends Microbiol. 2004 Oct;12(10):466-72</Citation><ArticleIdList><ArticleId IdType="pubmed">15381196</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Curr Opin Microbiol. 2004 Aug;7(4):412-9</Citation><ArticleIdList><ArticleId IdType="pubmed">15358261</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>N Engl J Med. 2003 May 15;348(20):1967-76</Citation><ArticleIdList><ArticleId IdType="pubmed">12690091</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Curr Pharm Des. 2006;12(35):4555-64</Citation><ArticleIdList><ArticleId IdType="pubmed">17168761</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Proc Natl Acad Sci U S A. 2004 Jul 6;101(27):10012-7</Citation><ArticleIdList><ArticleId IdType="pubmed">15226499</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Lancet. 2003 Oct 25;362(9393):1353-8</Citation><ArticleIdList><ArticleId IdType="pubmed">14585636</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>J Virol. 2000 Nov;74(22):10600-11</Citation><ArticleIdList><ArticleId IdType="pubmed">11044104</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Science. 2003 May 30;300(5624):1394-9</Citation><ArticleIdList><ArticleId IdType="pubmed">12730500</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Antiviral Res. 2006 Jun;70(2):39-50</Citation><ArticleIdList><ArticleId IdType="pubmed">16724398</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>Curr Pharm Des. 2006;12(35):4573-90</Citation><ArticleIdList><ArticleId IdType="pubmed">17168763</ArticleId></ArticleIdList></Reference></ReferenceList><ReferenceList><Reference><Citation>J Gen Virol. 2004 Jun;85(Pt 6):1717-1725</Citation><ArticleIdList><ArticleId IdType="pubmed">15166457</ArticleId></ArticleIdList></Reference></ReferenceList></PubmedData></pubmed></record>Pour manipuler ce document sous Unix (Dilib)
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