Phosphoramidate dinucleosides as hepatitis C virus polymerase inhibitors.
GC dinucleosides exhibiting a phosphoramidate internucleosidic linkage with neutral, amphiphile, positively or negatively charged side chains were synthesized. Their potential inhibitory effect on the hepatitis C virus (HCV) NS5B polymerase was evaluated in vitro and in HCV replicon containing cells. Whereas the amphiphile and the positively charged analogues were found to be inactive, the neutral (1) and the negatively charged (4) ones inhibited enzyme activity when tested as a diastereoisomeric mixture. The most potent inhibitor proved to be the Sp isomer of the 5'-thiophosphorylated dinucleotide bearing the carboxylic side chain (8) (IC 50 of 25 microM in vitro and an EC 50 of 9 microM in HCV subgenomic replicon). Molecular modeling suggests that the phosphoramidate dinucleoside (8) is stabilized in the active site by interactions with magnesium ions and lysine and arginine residues of the polymerase.
Zlatev
Ivan
I
Institut des Biomolécules Max Mousseron, UMR 5247 CNRS - Universite Montpellier 1 - Universite Montpellier 2, Place Eugene Bataillon, CC1704, 34095 Montpellier Cedex 5, France.
Dutartre
Hélène
H
Barvik
Ivan
I
Neyts
Johan
J
Canard
Bruno
B
Vasseur
Jean-Jacques
JJ
Alvarez
Karine
K
Morvan
François
F
D045744:Cell Line, Tumor; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D016679:Genome, Viral; D016174:Hepacivirus / Q000201:enzymology* / Q000235:genetics / Q000502:physiology; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D009705:Nucleosides / Q000737:chemistry / Q000494:pharmacology*; D009943:Organophosphorus Compounds / Q000737:chemistry*; D010084:Oxidation-Reduction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D017361:Viral Nonstructural Proteins / Q000037:antagonists & inhibitors*; D014779:Virus Replication / Q000187:drug effects; D012324:RNA-Dependent RNA Polymerase
D004791:Enzyme Inhibitors; D009705:Nucleosides; D009943:Organophosphorus Compounds; D017361:Viral Nonstructural Proteins; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase