Biochemical Characterization of the Active Anti-Hepatitis C Virus Metabolites of 2,6-Diaminopurine Ribonucleoside Prodrug Compared to Sofosbuvir and BMS-986094
| Autor: | Franck Amblard, Tony Whitaker, Sheida Amiralaei, Biing Lin, Jadd R. Shelton, Hongwang Zhang, Sijia Tao, Longhu Zhou, Robert A. Domaoal, Ahmed Khalil, Jong Hyun Cho, Steven J. Coats, Maryam Ehteshami, Tugba Ozturk, Xiao Lu, Richard A. Stanton, Sam S. Lee, Justin E. Suesserman, Raymond F. Schinazi |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Adenosine Transcription Genetic RNA Mitochondrial Hepatitis C virus Metabolite 030106 microbiology Guanosine Monophosphate Hepacivirus Viral Nonstructural Proteins Virus Replication medicine.disease_cause Antiviral Agents Cell Line 03 medical and health sciences chemistry.chemical_compound Transcription (biology) RNA polymerase medicine Humans Prodrugs Pharmacology (medical) NS5B Polymerase Pharmacology biology Chemistry DNA-Directed RNA Polymerases Prodrug Ribonucleoside Hepatitis C Infectious Diseases Biochemistry biology.protein RNA RNA Viral Ribonucleosides Sofosbuvir |
| Zdroj: | Antimicrobial Agents and Chemotherapy. 60:4659-4669 |
| ISSN: | 1098-6596 0066-4804 |
| DOI: | 10.1128/aac.00318-16 |
| Popis: | Ribonucleoside analog inhibitors (rNAI) target the hepatitis C virus (HCV) RNA-dependent RNA polymerase nonstructural protein 5B (NS5B) and cause RNA chain termination. Here, we expand our studies on β- d -2′- C -methyl-2,6-diaminopurine-ribonucleotide (DAPN) phosphoramidate prodrug 1 (PD1) as a novel investigational inhibitor of HCV. DAPN-PD1 is metabolized intracellularly into two distinct bioactive nucleoside triphosphate (TP) analogs. The first metabolite, 2′- C -methyl-GTP, is a well-characterized inhibitor of NS5B polymerase, whereas the second metabolite, 2′- C -methyl-DAPN-TP, behaves as an adenosine base analog. In vitro assays suggest that both metabolites are inhibitors of NS5B-mediated RNA polymerization. Additional factors, such as rNAI-TP incorporation efficiencies, intracellular rNAI-TP levels, and competition with natural ribonucleotides, were examined in order to further characterize the potential role of each nucleotide metabolite in vivo . Finally, we found that although both 2′- C -methyl-GTP and 2′- C -methyl-DAPN-TP were weak substrates for human mitochondrial RNA (mtRNA) polymerase (POLRMT) in vitro , DAPN-PD1 did not cause off-target inhibition of mtRNA transcription in Huh-7 cells. In contrast, administration of BMS-986094, which also generates 2′- C -methyl-GTP and previously has been associated with toxicity in humans, caused detectable inhibition of mtRNA transcription. Metabolism of BMS-986094 in Huh-7 cells leads to 87-fold higher levels of intracellular 2′- C -methyl-GTP than DAPN-PD1. Collectively, our data characterize DAPN-PD1 as a novel and potent antiviral agent that combines the delivery of two active metabolites. |
| Databáze: | OpenAIRE |
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