Resistance Profile of a Hepatitis C Virus RNA-Dependent RNA Polymerase Benzothiadiazine Inhibitor
| العنوان: | Resistance Profile of a Hepatitis C Virus RNA-Dependent RNA Polymerase Benzothiadiazine Inhibitor |
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| المؤلفون: | Baohua Gu, Robert T. Sarisky, Kevin J. Duffy, Adam T. Gates, Lester L. Gutshall, Tammy T. Nguyen, Victor K. Johnston |
| المصدر: | Antimicrobial Agents and Chemotherapy. 47:3525-3530 |
| بيانات النشر: | American Society for Microbiology, 2003. |
| سنة النشر: | 2003 |
| مصطلحات موضوعية: | DNA, Complementary, viruses, Hepatitis C virus, RNA-dependent RNA polymerase, Hepacivirus, Benzothiadiazines, medicine.disease_cause, Antiviral Agents, Cell Line, Viral Proteins, chemistry.chemical_compound, RNA polymerase, Drug Resistance, Viral, medicine, Humans, Pharmacology (medical), Replicon, Enzyme Inhibitors, Polymerase Gene, Gene, NS5B, Polymerase, Pharmacology, biology, biochemical phenomena, metabolism, and nutrition, RNA-Dependent RNA Polymerase, Virology, Molecular biology, Infectious Diseases, chemistry, Mutagenesis, Site-Directed, biology.protein, RNA, Viral |
| الوصف: | Recently, a benzo-1,2,4-thiadiazine antiviral agent (C 21 H 21 N 3 O 4 S; compound 4) was shown to be a potent, highly specific inhibitor of the primary catalytic enzyme of the hepatitis C virus (HCV) replicase complex. In this study, we selected for resistance to confirm the mechanism of action for compound 4 in HCV replicon cells. As expected, spontaneous mutations or fluidity in the HCV polymerase (NS5B) coding sequence occurred upon routine passage of the HCV replicon cells in the absence of compound 4. After 1 month of culture in the presence of 10 μM compound 4, or 20 times the 50% inhibitory concentration of the replicon, replicon cells were almost 20-fold less susceptible to compound 4. Twenty-one NS5B cDNA clones were generated from the resistant replicon cells. Five mutations in the 21 NS5B clones were present at frequencies higher than that of control replicon cells, and no clone contained more than a single mutation within the polymerase gene. RNA-dependent RNA polymerase studies using purified recombinant NS5B containing these single point mutations allowed the identification of residue 414 as sufficient for biochemical resistance to compound 4. Further, the contribution of this residue to confer cell-based resistance to compound 4 was validated using a stable recombinant mutant replicon cell line which harbors a methionine-to-threonine change at residue 414. The potential for additional mutations in other nonstructural genes of HCV to contribute to the resistance profile of compound 4 is discussed. |
| تدمد: | 1098-6596 0066-4804 |
| DOI: | 10.1128/aac.47.11.3525-3530.2003 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::fbfd6bc297392e4a33ee0229da99c35f https://doi.org/10.1128/aac.47.11.3525-3530.2003 |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....fbfd6bc297392e4a33ee0229da99c35f |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 10986596 00664804 |
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| DOI: | 10.1128/aac.47.11.3525-3530.2003 |