title: Targeted delivery of a triplex-forming oligonucleotide to hepatic stellate cells.
abstract: Liver fibrosis is characterized by abnormal accumulation of extracellular matrix (ECM), namely, fibrillar collagens in the hepatic stellate cells (HSCs). Earlier, we developed an antigene approach, using a type alpha1(I) collagen gene promoter specific triplex-forming oligonucleotide (TFO) to inhibit collagen gene expression. In this paper, to enhance overall delivery of TFOs to the liver and more specifically to HSCs, we synthesized mannose 6-phosphate-bovine serum albumin (M6P-BSA) by phosphorylating p-nitrophenyl-alpha-d-mannopyranoside, reducing its nitro group, and reacting it with thiophosgene to produce p-isothiocyanatophenyl-6-phospho-alpha-d-mannopyranoside (itcM6P) for conjugation with BSA. (33)P-TFO was conjugated with M6P-BSA via a disulfide bond, and the stability of the (M6P)(20)-BSA-TFO conjugate was determined. Following tail vein injection into rats, (M6P)(20)-BSA-(33)P-TFO rapidly cleared from the circulation and accumulated mainly in the liver. Almost 66% of the injected (M6P)(20)-BSA-(33)P-TFO accumulated in the liver at 30 min postinjection, which was significantly higher than that deposited after injection of (33)P-TFO. A large proportion of the injected (M6P)(20)-BSA-(33)P-TFO was taken up by the HSCs as evidenced by determination of radioactivity in the digested liver cells upon liver perfusion and separation on a Nycodenz gradient. Therefore, this TFO conjugate may be used for the treatment of liver fibrosis.
authors.affiliation: Department of Pharmaceutical, University of Tennessee Health Science Center, Memphis, Tennessee 38163, USA.
authors.forename: Zhaoyang
authors.initials: Z
authors.lastname: Ye
authors.forename: Kun
authors.initials: K
authors.lastname: Cheng
authors.forename: Ramareddy V
authors.initials: RV
authors.lastname: Guntaka
authors.forename: Ram I
authors.initials: RI
authors.lastname: Mahato
mesh_terms: D000818:Animals; D004247:DNA / Q000008:administration & dosage* / Q000138:chemical synthesis / Q000378:metabolism; D004220:Disulfides / Q000737:chemistry; D004337:Drug Carriers / Q000008:administration & dosage / Q000138:chemical synthesis / Q000378:metabolism; D018390:Gene Targeting / Q000379:methods; D066298:In Vitro Techniques; D008099:Liver / Q000166:cytology* / Q000378:metabolism*; D008297:Male; D008360:Mannosephosphates / Q000008:administration & dosage / Q000138:chemical synthesis; D009690:Nucleic Acid Conformation / Q000187:drug effects; D009838:Oligodeoxyribonucleotides / Q000008:administration & dosage* / Q000138:chemical synthesis / Q000378:metabolism; D010477:Perfusion; D051381:Rats; D017207:Rats, Sprague-Dawley; D012710:Serum Albumin, Bovine / Q000008:administration & dosage / Q000138:chemical synthesis; D013388:Succinimides / Q000008:administration & dosage / Q000138:chemical synthesis; D013873:Thionucleotides / Q000008:administration & dosage / Q000138:chemical synthesis / Q000378:metabolism
chemical_list: D004220:Disulfides; D004337:Drug Carriers; D008360:Mannosephosphates; D009838:Oligodeoxyribonucleotides; D013388:Succinimides; D013873:Thionucleotides; C051802:triplex DNA; D012710:Serum Albumin, Bovine; C018151:N-succinimidyl 3-(2-pyridyldithio)propionate; C027693:mannose-6-phosphate; D004247:DNA