Abstract
Resistant HCV variants carrying NS5B S282T mutation confer reduced sensitivity to sofosbuvir, the sole marketed NS5B polymerase inhibitor. On the basis of the finding that 2'-α-F-2'-β-C-methylcytidine 5'-triphosphate (8) was more potent than sofosbuvir's active metabolite on inhibition of both wild-type and S282T mutant polymerase, a dual-prodrug approach has been established. Twenty-nine phosphoramidates with N4-modified cytosine were designed, synthesized, and evaluated for anti-HCV activity. The results showed that compounds 4c-4e and 4m (EC50 = 0.19-0.25 μM) exhibited comparable potency to that of sofosbuvir (EC50 = 0.15 μM) on inhibition of wild-type replicons. Notably, 4c (EC50 = 0.366 μM) was 1.5-fold more potent than sofosbuvir (EC50 = 0.589 μM) on inhibition of S282T mutant replicons. In vitro metabolic studies disclosed the possible metabolic pathways of 4c. The toxicity study results indicated a good safety profile of 4c. Together, 4c-4e and 4m hold promise for drug development for the treatment of HCV infection, especially the resistant variants with NS5B S282T mutation.
MeSH terms
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Alanine / analogs & derivatives*
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Alanine / chemical synthesis
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Alanine / pharmacokinetics
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Alanine / pharmacology
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Animals
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Antiviral Agents / chemical synthesis
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Antiviral Agents / chemistry*
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Antiviral Agents / pharmacology
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Cell Line, Tumor
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Cytidine Monophosphate / analogs & derivatives*
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Cytidine Monophosphate / chemical synthesis
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Cytidine Monophosphate / pharmacokinetics
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Cytidine Monophosphate / pharmacology
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Dogs
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Female
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Hepacivirus / drug effects*
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Hepacivirus / genetics
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Humans
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Liver / metabolism
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Male
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Mutation
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Nucleotides / chemical synthesis*
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Nucleotides / pharmacokinetics
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Nucleotides / pharmacology
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Prodrugs / chemical synthesis*
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Prodrugs / pharmacokinetics
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Prodrugs / pharmacology
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RNA-Dependent RNA Polymerase / genetics
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Replicon
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Stereoisomerism
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Structure-Activity Relationship
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Viral Nonstructural Proteins / genetics
Substances
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Antiviral Agents
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Nucleotides
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Prodrugs
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Viral Nonstructural Proteins
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isopropyl 2-((((5-(4-butyramido-2-oxopyrimidin-1(2H)-yl)-4-fluoro-3-hydroxy-4-methyltetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)amino)propanoate
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NS-5 protein, hepatitis C virus
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RNA-Dependent RNA Polymerase
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Cytidine Monophosphate
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Alanine