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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<author><name sortKey="Ge, Feng" sort="Ge, Feng" uniqKey="Ge F" first="Feng" last="Ge">Feng Ge</name>
<affiliation><nlm:affiliation>Institute of Life and Health Engineering, Jinan University, Guangzhou, Guangdong, PR China. tgfeng@jnu.edu.cn</nlm:affiliation>
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<author><name sortKey="Xiong, Sheng" sort="Xiong, Sheng" uniqKey="Xiong S" first="Sheng" last="Xiong">Sheng Xiong</name>
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<author><name sortKey="Lin, Fu Sen" sort="Lin, Fu Sen" uniqKey="Lin F" first="Fu-Sen" last="Lin">Fu-Sen Lin</name>
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<author><name sortKey="Zhang, Zhi Ping" sort="Zhang, Zhi Ping" uniqKey="Zhang Z" first="Zhi-Ping" last="Zhang">Zhi-Ping Zhang</name>
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<author><name sortKey="Zhang, Xian En" sort="Zhang, Xian En" uniqKey="Zhang X" first="Xian-En" last="Zhang">Xian-En Zhang</name>
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<affiliation><nlm:affiliation>Institute of Life and Health Engineering, Jinan University, Guangzhou, Guangdong, PR China. tgfeng@jnu.edu.cn</nlm:affiliation>
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<author><name sortKey="Zhang, Zhi Ping" sort="Zhang, Zhi Ping" uniqKey="Zhang Z" first="Zhi-Ping" last="Zhang">Zhi-Ping Zhang</name>
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<term>Chlorocebus aethiops</term>
<term>Cricetinae</term>
<term>Drug Evaluation, Preclinical</term>
<term>Green Fluorescent Proteins (genetics)</term>
<term>Green Fluorescent Proteins (metabolism)</term>
<term>Humans</term>
<term>Replicon</term>
<term>SARS Virus (drug effects)</term>
<term>SARS Virus (genetics)</term>
<term>Severe Acute Respiratory Syndrome (drug therapy)</term>
<term>Severe Acute Respiratory Syndrome (virology)</term>
<term>Small Molecule Libraries (pharmacology)</term>
<term>Vero Cells</term>
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<term>Small Molecule Libraries</term>
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<front><div type="abstract" xml:lang="en">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.</div>
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<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>
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