title: Novel nucleoside analogues targeting HCV replication through an NS5A-dependent inhibition mechanism.
abstract: A series of new tricyclic nucleosides were synthesized and evaluated as hepatitis C virus (HCV) replication inhibitors. Initial screening in a HCV replicon system, derived from a genotype 1b isolate, identified 9-benzylamino-3-(β-D-ribofuranosyl)-3H-imidazo[4',5':5,6]pyrido[2,3-b]pyrazine (15d) as the most potent analogue. Comparative assessment of 15d activity against HCV full-length viruses or subgenomic replicons derived from genotypes 1 to 4 revealed a specificity of the compound for genotypes 1 and 3. Surprisingly, resistance mutations selected against 15d were mapped to domains II and III of the non-structural protein 5A (NS5A), but not to the RNA-dependent RNA polymerase residing in NS5B. These results argue that compound 15d might represent a lead for the development of a novel class of NS5A inhibitors.
authors.affiliation: Division of Pharmaceutical Chemistry, Department of Pharmacy, National and Kapodistrian University of Athens, Athens, Greece.
authors.forename: Nikolaos
authors.initials: N
authors.lastname: Lougiakis
authors.affiliation: Molecular Virology Laboratory, Hellenic Pasteur Institute, Athens, Greece.
authors.forename: Efseveia
authors.initials: E
authors.lastname: Frakolaki
authors.affiliation: Division of Pharmaceutical Chemistry, Department of Pharmacy, National and Kapodistrian University of Athens, Athens, Greece.
authors.forename: Panagiota
authors.initials: P
authors.lastname: Karmou
authors.affiliation: Division of Pharmaceutical Chemistry, Department of Pharmacy, National and Kapodistrian University of Athens, Athens, Greece.
authors.forename: Nicole
authors.identifier: http://orcid.org/0000-0003-3272-759X
authors.initials: N
authors.lastname: Pouli
authors.affiliation: Division of Pharmaceutical Chemistry, Department of Pharmacy, National and Kapodistrian University of Athens, Athens, Greece.
authors.forename: Panagiotis
authors.initials: P
authors.lastname: Marakos
authors.affiliation: Department of Infectious Diseases, Molecular Virology, University of Heidelberg, Heidelberg, Germany.
authors.forename: Vanesa
authors.initials: V
authors.lastname: Madan
authors.affiliation: Department of Infectious Diseases, Molecular Virology, University of Heidelberg, Heidelberg, Germany.|German Center for Infection Research, Heidelberg University, Heidelberg, Germany.|Division of Virus-Associated Carcinogenesis, German Cancer Research Center (DKFZ), Heidelberg, Germany.
authors.forename: Ralf
authors.initials: R
authors.lastname: Bartenschlager
authors.affiliation: Molecular Virology Laboratory, Hellenic Pasteur Institute, Athens, Greece.
authors.forename: Niki
authors.initials: N
authors.lastname: Vassilaki
mesh_terms: D000998:Antiviral Agents / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology; D002460:Cell Line; D024882:Drug Resistance, Viral / Q000187:drug effects; D017930:Genes, Reporter; D005838:Genotype; D016174:Hepacivirus / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D006575:Heterocyclic Compounds, 3-Ring / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008856:Microscopy, Fluorescence; D009705:Nucleosides / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology*; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D012367:RNA, Viral / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D017423:Sequence Analysis, RNA; D017361:Viral Nonstructural Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D014779:Virus Replication / Q000187:drug effects*; D012324:RNA-Dependent RNA Polymerase
chemical_list: D000998:Antiviral Agents; D006575:Heterocyclic Compounds, 3-Ring; D009705:Nucleosides; D012367:RNA, Viral; D017361:Viral Nonstructural Proteins; C000623128:9-benzylamino-3-(ribofuranosyl)-3H-imidazo(4',5'-5,6)pyrido(2,3-b)pyrazine; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase
keywords: N-ribonucleosides; NS5A domains II, III; antiviral activity; imidazo[4′,5′:5,6]pyrido[2,3-b]pyrazine; nucleoside scaffold; resistance mutations