title: Covalent protein-oligonucleotide conjugates for efficient delivery of antisense molecules.
abstract: Antisense oligonucleotides have been covalently attached to asialoglycoprotein (ASGP) via disulfide bond conjugation chemistry. These conjugates were characterized extensively by an array of chemical, chromatographic, and spectroscopic means. Multiple (approximately six) oligonucleotides can be conjugated to each ASGP molecule. The molecular conjugates were used to deliver antisense oligonucleotides complementary to the mRNA of the interleukin 6 signal transduction protein (gp130) to modulate the acute phase response of hepatoma (HepG2) cells in vitro. These conjugates were biologically active, as measured by inhibition of the cytokine-stimulated up-regulation of the acute phase protein haptoglobin. The level of inhibition was comparable to that found with previous technology featuring noncovalent complexes of ASGP-poly(L-lysine) and oligonucleotide. Because of the ability to control the stoichiometry of the conjugate and its unimolecular nature (as opposed to bimolecular for the noncovalent conjugates), this methodology holds great promise for further development and application.
authors.affiliation: Center for Engineering in Medicine, Massachusetts General Hospital, Shriners Burns Institute, Boston 02114, USA.
authors.forename: S B
authors.initials: SB
authors.lastname: Rajur
authors.forename: C M
authors.initials: CM
authors.lastname: Roth
authors.forename: J R
authors.initials: JR
authors.lastname: Morgan
authors.forename: M L
authors.initials: ML
authors.lastname: Yarmush
mesh_terms: D001212:Asialoglycoproteins / Q000008:administration & dosage* / Q000138:chemical synthesis / Q000737:chemistry*; D006528:Carcinoma, Hepatocellular / Q000378:metabolism; D003432:Cross-Linking Reagents / Q000737:chemistry; D004220:Disulfides / Q000737:chemistry; D004337:Drug Carriers; D016503:Drug Delivery Systems; D006801:Humans; D016376:Oligonucleotides, Antisense / Q000008:administration & dosage* / Q000138:chemical synthesis / Q000737:chemistry*; D011725:Pyridines / Q000737:chemistry; D013386:Succinates / Q000737:chemistry; D014407:Tumor Cells, Cultured / Q000187:drug effects
chemical_list: D001212:Asialoglycoproteins; D003432:Cross-Linking Reagents; D004220:Disulfides; D004337:Drug Carriers; D016376:Oligonucleotides, Antisense; D011725:Pyridines; D013386:Succinates; C109849:sulfosuccinimidyl 6-(3'-(2-pyridyldithio)propionamido)hexanoate