title: Phosphoramidate dinucleosides as hepatitis C virus polymerase inhibitors.
abstract: 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.
authors.affiliation: Institut des Biomolécules Max Mousseron, UMR 5247 CNRS - Universite Montpellier 1 - Universite Montpellier 2, Place Eugene Bataillon, CC1704, 34095 Montpellier Cedex 5, France.
authors.forename: Ivan
authors.initials: I
authors.lastname: Zlatev
authors.forename: Hélène
authors.initials: H
authors.lastname: Dutartre
authors.forename: Ivan
authors.initials: I
authors.lastname: Barvik
authors.forename: Johan
authors.initials: J
authors.lastname: Neyts
authors.forename: Bruno
authors.initials: B
authors.lastname: Canard
authors.forename: Jean-Jacques
authors.initials: JJ
authors.lastname: Vasseur
authors.forename: Karine
authors.initials: K
authors.lastname: Alvarez
authors.forename: François
authors.initials: F
authors.lastname: Morvan
mesh_terms: 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
chemical_list: D004791:Enzyme Inhibitors; D009705:Nucleosides; D009943:Organophosphorus Compounds; D017361:Viral Nonstructural Proteins; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase