title: High affinity peptide inhibitors of the hepatitis C virus NS3-4A protease refractory to common resistant mutants.
abstract: Hepatitis C virus (HCV) NS3-4A protease is essential for viral replication. All current small molecular weight drugs against NS3-4A are substrate peptidomimetics that have a similar binding and resistance profile. We developed inhibitory peptides (IPs) capping the active site and binding via a novel "tyrosine" finger at an alternative NS3-4A site that is of particular interest for further HCV drug development. The peptides are not cleaved due to a combination of geometrical constraints and impairment of the oxyanion hole function. Selection and optimization through combinatorial phagemid display, protein crystallography, and further modifications resulted in a 32-amino acid peptide with a K(i) of 0.53 nm. Inhibition of viral replication in cell culture was demonstrated by fusion to a cell-penetrating peptide. Negligible susceptibility to known (A156V and R155K) resistance mutations of the NS3-4A protease was observed. This work shows for the first time that antiviral peptides can target an intracellular site and reveals a novel druggable site on the HCV protease.
authors.affiliation: Research Group Directed Evolution, Helmholtz Centre for Infection Research (HZI), 38124 Braunschweig, Germany.
authors.forename: Jonas
authors.initials: J
authors.lastname: Kügler
authors.forename: Stefan
authors.initials: S
authors.lastname: Schmelz
authors.forename: Juliane
authors.initials: J
authors.lastname: Gentzsch
authors.forename: Sibylle
authors.initials: S
authors.lastname: Haid
authors.forename: Erik
authors.initials: E
authors.lastname: Pollmann
authors.forename: Joop
authors.initials: J
authors.lastname: van den Heuvel
authors.forename: Raimo
authors.initials: R
authors.lastname: Franke
authors.forename: Thomas
authors.initials: T
authors.lastname: Pietschmann
authors.forename: Dirk W
authors.initials: DW
authors.lastname: Heinz
authors.forename: John
authors.initials: J
authors.lastname: Collins
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology; D001665:Binding Sites; D002352:Carrier Proteins / Q000737:chemistry*; D057846:Cell-Penetrating Peptides / Q000737:chemistry; D003461:Crystallography / Q000379:methods; D015195:Drug Design; D024882:Drug Resistance, Viral / Q000187:drug effects / Q000235:genetics; D006801:Humans; D020128:Inhibitory Concentration 50; D047908:Intracellular Signaling Peptides and Proteins; D007700:Kinetics; D008958:Models, Molecular; D008968:Molecular Conformation; D009154:Mutation; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry; D012997:Solvents / Q000737:chemistry; D017361:Viral Nonstructural Proteins / Q000737:chemistry*; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases
chemical_list: D000998:Antiviral Agents; D002352:Carrier Proteins; D057846:Cell-Penetrating Peptides; D047908:Intracellular Signaling Peptides and Proteins; D019151:Peptide Library; D010455:Peptides; D012997:Solvents; D017361:Viral Nonstructural Proteins; C084422:NS3 protein, hepatitis C virus; D012697:Serine Endopeptidases; D043583:Nucleoside-Triphosphatase; D053487:DEAD-box RNA Helicases; D000085242:Viral Proteases; C455041:NS4A cofactor peptide, Hepatitis C virus