Inhibition of the replication of a hepatitis C virus-like RNA template by interferon and 3'-deoxycytidine.
The development of low molecular weight inhibitors of hepatitis C virus (HCV) replication has been hindered by the lack of a good cell-based system that models the entire HCV replication cycle. To date the only two therapies approved for the treatment of HCV infection are interferon (IFN)-alpha and the nucleoside analogue, ribavirin. We have created a cell-based system that allows for the accurate quantification of the replication of an HCV-like RNA template by proteins that are encoded for by the HCV genome. The system consists of a cell line that constitutively produces luciferase in response to the production of functional HCV replicative proteins. The 293B4alpha cell line has been formatted into a semi-high throughput, cell-based screen for inhibitors of HCV replication. When these cells were treated with either IFN-alpha or -beta, luciferase production decreased in a dose-responsive manner. Counterscreening these molecules in another cell line, 293SVLuc, in which luciferase production in not dependent the presence of functional HCV proteins, showed that the inhibition of luciferase in the 293B4alpha cell line was due to inhibition of the replication of the HCV-like RNA template and not anti-cellular or -luciferase activity. Moreover, when the 293B4alpha cell line was treated with the ribonucleoside analogue, 3'-deoxycytidine, luciferase decreased in a dose-responsive manner. 3'-deoxyguanosine and 3'-deoxyuridine did not inhibit luciferase production and 3'-deoxyadenosine was too cytotoxic to determine if it had any anti-HCV activity.
King
Robert W
RW
The Experimental Station, Bristol-Myers Squibb, Wilmington, Del., USA. robert.king@bms.com
Zecher
Marianne
M
Jefferies
Matthew W
MW
D000998:Antiviral Agents / Q000494:pharmacology*; D002460:Cell Line / Q000821:virology; D003839:Deoxyadenosines / Q000494:pharmacology; D003841:Deoxycytidine / Q000031:analogs & derivatives / Q000494:pharmacology*; D003849:Deoxyguanosine / Q000031:analogs & derivatives / Q000494:pharmacology; D003857:Deoxyuridine / Q000031:analogs & derivatives / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D015195:Drug Design; D004353:Drug Evaluation, Preclinical; D017930:Genes, Reporter; D006367:HeLa Cells / Q000821:virology; D016174:Hepacivirus / Q000187:drug effects* / Q000235:genetics / Q000502:physiology; D022781:Hepatocytes / Q000821:virology; D006801:Humans; D016898:Interferon-alpha / Q000494:pharmacology*; D016899:Interferon-beta / Q000494:pharmacology*; D007668:Kidney; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D012367:RNA, Viral / Q000096:biosynthesis* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D012254:Ribavirin / Q000494:pharmacology; D013698:Templates, Genetic; D014779:Virus Replication / Q000187:drug effects*
C475627:3'-deoxyuridine; D000998:Antiviral Agents; D003839:Deoxyadenosines; D016898:Interferon-alpha; D012367:RNA, Viral; D011993:Recombinant Fusion Proteins; D003841:Deoxycytidine; C475626:3-deoxyguanosine; D012254:Ribavirin; C475625:3'-deoxycytidine; D016899:Interferon-beta; D008156:Luciferases; D003849:Deoxyguanosine; C058120:cordycepin; D003857:Deoxyuridine