Novel receptor-mediated gene delivery system comprising plasmid/protamine/sugar-containing polyanion ternary complex.
Poly(ethylene glycol) (PEG) derivative having both carboxylic acid-, and lactose-side chains (Lac-PEG-C) deposited onto the surface of DNA/protamine (PRT) complex, and the self-assembled ternary complex was obtained. The diameter of the complexes was 180-200 nm, and they showed good size stability even in the high ionic strength solutions. Lac-PEG-C coating reduced their surface electric potential, and effectively avoided the albumin-induced aggregation. DNA/PRT/Lac-PEG-C complex did not coagulate the red blood cells, and their cytotoxicity evaluated by WST-1 was very low. Lac-PEG-C added to the plasmid/PRT complex prior to the incubation with HepG2 cells extremely enhanced the gene-expression, and by the plasmid/PRT/Lac-PEG-C complex prepared at 1:1.5:8 in weight, 56-fold higher expression of luciferase than that without Lac-PEG-C was observed. The treatment with asialofetuin or phenylarsine oxide evidently interfered with the gene-expression. The high gene expression by the plasmid/PRT/Lac-PEG-C ternary complex on the hepatocyte would be attributed to the asialoglycoprotein receptor-mediated endocytosis.
Faculty of Pharmaceutical Sciences, Teikyo University, Sagamiko, Kanagawa 199-0195, Japan.
Kazuo
K
Maruyama
Fumito
F
Iwasaki
Tomoko
T
Takizawa
Hironobu
H
Yanagie
Takuro
T
Niidome
Etsuko
E
Yamada
Tomoko
T
Ito
Yoshiyuki
Y
Koyama
D006528:Carcinoma, Hepatocellular / Q000235:genetics* / Q000473:pathology; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D020099:Coated Materials, Biocompatible / Q000737:chemistry*; D016503:Drug Delivery Systems / Q000379:methods*; D018390:Gene Targeting / Q000379:methods; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D006801:Humans; D007785:Lactose / Q000737:chemistry*; D008113:Liver Neoplasms / Q000235:genetics / Q000473:pathology; D008422:Materials Testing; D010957:Plasmids / Q000008:administration & dosage* / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry*; D011479:Protamines / Q000737:chemistry*; D014162:Transfection / Q000379:methods
D020099:Coated Materials, Biocompatible; D011479:Protamines; D011092:Polyethylene Glycols; D007785:Lactose