title: Improvement of hepatocyte-specific gene expression by a targeted colchicine prodrug.
abstract: Colchicine, an established tubulin inhibitor, interferes with the trafficking of endocytotic vesicles and thereby promotes the escape of lysosome-entrapped compounds. To improve its potency and cell specificity, a targeted prodrug of colchicine was synthesized by conjugation to a high-affinity ligand (di-N(alpha),N(epsilon)-(5-(2-acetamido-2-deoxy-beta-D-galactopyranosyloxy)pentanomido)lysine, K(GalNAc)(2)) for the asialoglycoprotein receptor on parenchymal liver cells. The resulting colchicine-K(GalNAc)(2) conjugate bound to this receptor with an affinity of 4.5 nM. Confocal microscopy studies confirmed rapid uptake and receptor dependency of a prodrug conjugated with fluorescein isothiocyanate. Colchicine-K(GalNAc)(2) substantially increased the transfection efficiency of polyplexed DNA in parenchymal liver cells in a concentration- and receptor-dependent fashion. Colchicine-K(GalNAc)(2) was found to enhance the transfection efficiency by 50-fold at 1 nM, whereas the parental colchicine was ineffective. In conclusion, this nontoxic colchicine-K(GalNAc)(2) conjugate can be a useful tool to improve the transfection efficiency of hepatic nonviral gene transfer vehicles.
authors.affiliation: Division of Biopharmaceutics, Leiden/Amsterdam Center for Drug Research, Gorlaeus Laboratories, Leiden University, P.O. Box 9502, 2300 RA Leiden, The Netherlands. rossenbe@lacdr.leidenuniv.nl
authors.forename: Sabine M W
authors.initials: SM
authors.lastname: van Rossenberg
authors.forename: Karen M
authors.initials: KM
authors.lastname: Sliedregt-Bol
authors.forename: Gerben
authors.initials: G
authors.lastname: Koning
authors.forename: Hans
authors.initials: H
authors.lastname: van den Elst
authors.forename: Theo J C
authors.initials: TJ
authors.lastname: van Berkel
authors.forename: Jacques H
authors.initials: JH
authors.lastname: van Boom
authors.forename: Gijs A
authors.initials: GA
authors.lastname: van der Marel
authors.forename: Erik A L
authors.initials: EA
authors.lastname: Biessen
mesh_terms: D000818:Animals; D037263:Asialoglycoprotein Receptor / Q000737:chemistry / Q000378:metabolism; D003078:Colchicine / Q000031:analogs & derivatives* / Q000494:pharmacology*; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects*; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism*; D008099:Liver / Q000166:cytology / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal; D011355:Prodrugs / Q000138:chemical synthesis / Q000378:metabolism / Q000494:pharmacology* / Q000633:toxicity; D011869:Radioligand Assay; D018675:Toxicity Tests / Q000379:methods; D014162:Transfection
chemical_list: D037263:Asialoglycoprotein Receptor; D011355:Prodrugs; D016650:Fluorescein-5-isothiocyanate; D003078:Colchicine