Abstract |
Several members of the widespread alphavirus group are pathogenic, but no therapy is available to treat these RNA virus infections. We report here a quantitative assay to screen for inhibitors of Semliki Forest virus (SFV) replication, and demonstrate the effects of 29 nucleosides on SFV and Sindbis virus replication. The anti-SFV assay developed is based on a SFV strain containing Renilla luciferase inserted after the nsP3 coding region, yielding a marker virus in which the luciferase is cleaved out during polyprotein processing. The reporter-gene assay was miniaturized, automated and validated, resulting in a Z' value of 0.52. [3H] uridine labeling for 1 h at the maximal viral RNA synthesis time point was used as a comparative method. Anti-SFV screening and counter-screening for cell viability led to the discovery of several new SFV inhibitors. 3'-amino-3'-deoxyadenosine was the most potent inhibitor in this set, with an IC50 value of 18 microM in the reporter-gene assay and 2 microM in RNA synthesis rate detection. Besides the 3'-substituted analogues, certain N6-substituted nucleosides had similar IC50 values for both SFV and Sindbis replication, suggesting the applicability of this methodology to alphaviruses in general.
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Authors | Leena Pohjala, Vladimir Barai, Alex Azhayev, Seppo Lapinjoki, Tero Ahola |
Journal | Antiviral research
(Antiviral Res)
Vol. 78
Issue 3
Pg. 215-22
(Jun 2008)
ISSN: 0166-3542 [Print] Netherlands |
PMID | 18294708
(Publication Type: Evaluation Study, Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- Antiviral Agents
- Deoxyadenosines
- Nucleosides
- RNA, Viral
- 3'-amino-3'-deoxyadenosine
- Luciferases
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Topics |
- Alphavirus
(drug effects, genetics, physiology)
- Animals
- Antiviral Agents
(chemistry, pharmacology)
- Cell Line
- Cricetinae
- Deoxyadenosines
(chemistry, pharmacology)
- Genes, Reporter
- Inhibitory Concentration 50
- Luciferases
(genetics, metabolism)
- Microbial Sensitivity Tests
(methods)
- Nucleosides
(chemistry, pharmacology)
- RNA, Viral
(biosynthesis)
- Semliki forest virus
(drug effects, genetics, physiology)
- Sindbis Virus
(drug effects, genetics, physiology)
- Virus Replication
(drug effects)
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