title: Hepatocyte targeting and intracellular copper chelation by a thiol-containing glycocyclopeptide.
abstract: Metal overload plays an important role in several diseases or intoxications, like in Wilson's disease, a major genetic disorder of copper metabolism in humans. To efficiently and selectively decrease copper concentration in the liver that is highly damaged, chelators should be targeted at the hepatocytes. In the present work, we synthesized a molecule able to both lower intracellular copper, namely Cu(I), and target hepatocytes, combining within the same structure a chelating unit and a carbohydrate recognition element. A cyclodecapeptide scaffold displaying a controlled conformation with two independent faces was chosen to introduce both units. One face displays a cluster of carbohydrates to ensure an efficient recognition of the asialoglycoprotein receptors, expressed on the surface of hepatocytes. The second face is devoted to metal ion complexation thanks to the thiolate functions of two cysteine side-chains. To obtain a chelator that is active only once inside the cells, the two thiol functions were oxidized in a disulfide bridge to afford the glycopeptide P(3). Two simple cyclodecapeptides modeling the reduced and complexing form of P(3) in cells proved a high affinity for Cu(I) and a high selectivity with respect to Zn(II). As expected, P(3) becomes an efficient Cu(I) chelator in the presence of glutathione that mimics the intracellular reducing environment. Finally, cellular uptake and ability to lower intracellular copper were demonstrated in hepatic cell lines, in particular in WIF-B9, making P(3) a good candidate to fight copper overload in the liver.
authors.affiliation: INAC, Service de Chimie Inorganique et Biologique (UMR_E 3 CEA UJF, FRE CNRS 3200), Commissariat à l'Energie Atomique, 17 Rue des Martyrs, 38054 Grenoble Cedex, France.
authors.forename: Anaïs M
authors.initials: AM
authors.lastname: Pujol
authors.forename: Martine
authors.initials: M
authors.lastname: Cuillel
authors.forename: Olivier
authors.initials: O
authors.lastname: Renaudet
authors.forename: Colette
authors.initials: C
authors.lastname: Lebrun
authors.forename: Peggy
authors.initials: P
authors.lastname: Charbonnier
authors.forename: Doris
authors.initials: D
authors.lastname: Cassio
authors.forename: Christelle
authors.initials: C
authors.lastname: Gateau
authors.forename: Pascal
authors.initials: P
authors.lastname: Dumy
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Mintz
authors.forename: Pascale
authors.initials: P
authors.lastname: Delangle
mesh_terms: D002478:Cells, Cultured; D002614:Chelating Agents / Q000737:chemistry* / Q000378:metabolism; D003300:Copper / Q000737:chemistry* / Q000378:metabolism; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D015394:Molecular Structure; D010456:Peptides, Cyclic / Q000737:chemistry* / Q000378:metabolism / Q000493:pharmacokinetics; D013438:Sulfhydryl Compounds / Q000737:chemistry* / Q000378:metabolism
chemical_list: D002614:Chelating Agents; D010456:Peptides, Cyclic; D013438:Sulfhydryl Compounds; D003300:Copper