NS5B is the RNA-dependent RNA polymerase that catalyzes the replication of the hepatitis C virus genome. It is a major target for antiviral drugs including nucleoside analogs, such as the prodrugs mericitabine and sofosbuvir, which get metabolized to 2?-fluoro-2?C-methylcytidine-5?-triphosphate and 2?-fluoro-2?C-methyluridine-5?-triphosphate, respectively. These analogs act as chain terminators after they are incorporated during RNA synthesis. Recently, it has been shown that NS5B can efficiently remove chain terminators by a nucleotide-mediated excision reaction that rescues RNA synthesis. In this study, we use transient-state kinetics to understand the efficiency of inhibition for five nucleoside analogs. We show that CTP analogs are readily incorporated into a growing primer by NS5B but are also efficiently excised. In contrast, although UMP analogs are more slowly incorporated, the excision of UMP is slow and inefficient, and modifications to the 2?-carbon of the UTP ribose ring further decreased rates of excision to an undetectable level. Taken together, these data suggest that the clinical effectiveness of sofosbuvir is largely a function of being intractable to nucleotide-mediated excision compared with similar nucleoside analogs.
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P Angeletti SpA, Merck Res Labs, Ist Ric Biol Mol, Via Pontina Km 30,000, I-00040 Pomezia, ItalyP Angeletti SpA, Merck Res Labs, Ist Ric Biol Mol, Via Pontina Km 30,000, I-00040 Pomezia, Italy
Colarusso, Stefania
Attenni, Barbara
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P Angeletti SpA, Merck Res Labs, Ist Ric Biol Mol, Via Pontina Km 30,000, I-00040 Pomezia, ItalyP Angeletti SpA, Merck Res Labs, Ist Ric Biol Mol, Via Pontina Km 30,000, I-00040 Pomezia, Italy
Attenni, Barbara
Avolio, Salvatore
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P Angeletti SpA, Merck Res Labs, Ist Ric Biol Mol, Via Pontina Km 30,000, I-00040 Pomezia, ItalyP Angeletti SpA, Merck Res Labs, Ist Ric Biol Mol, Via Pontina Km 30,000, I-00040 Pomezia, Italy
Avolio, Salvatore
Malancona, Savina
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P Angeletti SpA, Merck Res Labs, Ist Ric Biol Mol, Via Pontina Km 30,000, I-00040 Pomezia, ItalyP Angeletti SpA, Merck Res Labs, Ist Ric Biol Mol, Via Pontina Km 30,000, I-00040 Pomezia, Italy
Malancona, Savina
Harper, Steven
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P Angeletti SpA, Merck Res Labs, Ist Ric Biol Mol, Via Pontina Km 30,000, I-00040 Pomezia, ItalyP Angeletti SpA, Merck Res Labs, Ist Ric Biol Mol, Via Pontina Km 30,000, I-00040 Pomezia, Italy
Harper, Steven
Altamura, Sergio
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P Angeletti SpA, Merck Res Labs, Ist Ric Biol Mol, Via Pontina Km 30,000, I-00040 Pomezia, ItalyP Angeletti SpA, Merck Res Labs, Ist Ric Biol Mol, Via Pontina Km 30,000, I-00040 Pomezia, Italy
Altamura, Sergio
Koch, Uwe
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P Angeletti SpA, Merck Res Labs, Ist Ric Biol Mol, Via Pontina Km 30,000, I-00040 Pomezia, ItalyP Angeletti SpA, Merck Res Labs, Ist Ric Biol Mol, Via Pontina Km 30,000, I-00040 Pomezia, Italy
Koch, Uwe
Narjes, Frank
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P Angeletti SpA, Merck Res Labs, Ist Ric Biol Mol, Via Pontina Km 30,000, I-00040 Pomezia, ItalyP Angeletti SpA, Merck Res Labs, Ist Ric Biol Mol, Via Pontina Km 30,000, I-00040 Pomezia, Italy
机构:
Abbott Labs, Dept R4CQ, Global Pharmaceut Res & Dev, Abbott Pk, IL 60064 USAAbbott Labs, Dept R4CQ, Global Pharmaceut Res & Dev, Abbott Pk, IL 60064 USA
Middleton, T.
Lim, H. B.
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Abbott Labs, Dept R4CQ, Global Pharmaceut Res & Dev, Abbott Pk, IL 60064 USAAbbott Labs, Dept R4CQ, Global Pharmaceut Res & Dev, Abbott Pk, IL 60064 USA
Lim, H. B.
Montgomery, D.
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Abbott Labs, Dept R4CQ, Global Pharmaceut Res & Dev, Abbott Pk, IL 60064 USAAbbott Labs, Dept R4CQ, Global Pharmaceut Res & Dev, Abbott Pk, IL 60064 USA
Montgomery, D.
Rockway, T.
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Abbott Labs, Dept R4CQ, Global Pharmaceut Res & Dev, Abbott Pk, IL 60064 USAAbbott Labs, Dept R4CQ, Global Pharmaceut Res & Dev, Abbott Pk, IL 60064 USA
Rockway, T.
Liu, D.
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Abbott Labs, Dept R4CQ, Global Pharmaceut Res & Dev, Abbott Pk, IL 60064 USAAbbott Labs, Dept R4CQ, Global Pharmaceut Res & Dev, Abbott Pk, IL 60064 USA
Liu, D.
Klein, L.
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Abbott Labs, Dept R4CQ, Global Pharmaceut Res & Dev, Abbott Pk, IL 60064 USAAbbott Labs, Dept R4CQ, Global Pharmaceut Res & Dev, Abbott Pk, IL 60064 USA
Klein, L.
Qin, W.
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Abbott Labs, Dept R4CQ, Global Pharmaceut Res & Dev, Abbott Pk, IL 60064 USAAbbott Labs, Dept R4CQ, Global Pharmaceut Res & Dev, Abbott Pk, IL 60064 USA
Qin, W.
Harlan, J. E.
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Abbott Labs, Dept R4CQ, Global Pharmaceut Res & Dev, Abbott Pk, IL 60064 USAAbbott Labs, Dept R4CQ, Global Pharmaceut Res & Dev, Abbott Pk, IL 60064 USA
Harlan, J. E.
Kati, W. M.
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Abbott Labs, Dept R4CQ, Global Pharmaceut Res & Dev, Abbott Pk, IL 60064 USAAbbott Labs, Dept R4CQ, Global Pharmaceut Res & Dev, Abbott Pk, IL 60064 USA
Kati, W. M.
Molla, A.
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Abbott Labs, Dept R4CQ, Global Pharmaceut Res & Dev, Abbott Pk, IL 60064 USAAbbott Labs, Dept R4CQ, Global Pharmaceut Res & Dev, Abbott Pk, IL 60064 USA