In vitro and in vivo gene transfer by an optimized alpha-cyclodextrin conjugate with polyamidoamine dendrimer.
The purpose of the present study is to optimize the structure of the polyamidoamine starburst dendrimer (dendrimer) conjugate with alpha-cyclodextrin (alpha-CDE conjugate) as a nonviral vector. alpha-CDE conjugates of dendrimer (generation 3, G3) with various average degrees of substitution (DS) of alpha-CyD of 1.1, 2.4, and 5.4 were prepared. alpha-CDE conjugates formed the complexes with pDNA, resulting in a change of the particle sizes of pDNA complexes, but the distinction of physicochemical properties among their vector/pDNA complexes was only very slight. The membrane-disruptive ability of alpha-CDE conjugates on liposomes encapsulating calcein and their cytotoxicity to NIH3T3 and HepG2 increased with an increase in the DS value of alpha-CyD. In vitro gene transfer activity of alpha-CDE conjugates in both NIH3T3 and HepG2 cells augmented as the charge ratio (vector/pDNA) increased, and the activity of alpha-CDE conjugate (DS 2.4) was the highest at higher charge ratios among dendrimer (G3), the three alpha-CDE conjugates, and TransFast. After intravenous administration of pDNA complexes in mice, alpha-CDE conjugate (DS 2.4) delivered pDNA more efficiently in spleen, liver, and kidney, compared with dendrimer and other alpha-CDE conjugates (DS 1.1 and 5.4). The potential use of alpha-CDE conjugate (G3, DS 2.4) could be expected as a nonviral vector in vitro and in vivo, and these data may be useful for design of alpha-CyD conjugates with other nonviral vectors.
Kihara
Fumihiro
F
Faculty of Pharmaceutical Sciences, Kumamoto University, 5-1 Oe-honmachi, Japan.
Arima
Hidetoshi
H
Tsutsumi
Toshihito
T
Hirayama
Fumitoshi
F
Uekama
Kaneto
K
D016475:3T3 Cells; D000818:Animals; D002470:Cell Survival; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D003505:Cyclodextrins / Q000138:chemical synthesis / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D018014:Gene Transfer Techniques; D008081:Liposomes; D051379:Mice; D008807:Mice, Inbred BALB C; D009757:Nylons / Q000737:chemistry*; D010316:Particle Size; D010713:Phosphatidylcholines; D011073:Polyamines / Q000737:chemistry*; D014162:Transfection; D047391:alpha-Cyclodextrins
D003505:Cyclodextrins; D008081:Liposomes; D009757:Nylons; D010713:Phosphatidylcholines; D011073:Polyamines; D047391:alpha-Cyclodextrins; C032613:alpha-cyclodextrin