Kihara Fumihiro, Arima Hidetoshi, Tsutsumi Toshihito, Hirayama Fumitoshi, Uekama Kaneto
Faculty of Pharmaceutical Sciences, Kumamoto University, 5-1 Oe-honmachi, Japan.
Bioconjug Chem. 2003 Mar-Apr;14(2):342-50. doi: 10.1021/bc025613a.
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.
本研究的目的是优化聚酰胺-胺型树枝状大分子（树枝状聚合物）与α-环糊精的共轭物（α-CDE共轭物）作为非病毒载体的结构。制备了树枝状聚合物（第3代，G3）与α-CyD平均取代度（DS）分别为1.1、2.4和5.4的各种α-CDE共轭物。α-CDE共轭物与pDNA形成复合物，导致pDNA复合物的粒径发生变化，但其载体/pDNA复合物之间的物理化学性质差异非常小。α-CDE共轭物对包裹钙黄绿素的脂质体的膜破坏能力及其对NIH3T3和HepG2细胞的细胞毒性随着α-CyD的DS值增加而增强。α-CDE共轭物在NIH3T3和HepG2细胞中的体外基因转移活性随着电荷比（载体/pDNA）的增加而增强，并且在树枝状聚合物（G3）、三种α-CDE共轭物和TransFast中，α-CDE共轭物（DS 2.4）在较高电荷比时活性最高。在小鼠静脉注射pDNA复合物后，与树枝状聚合物和其他α-CDE共轭物（DS 1.1和5.4）相比，α-CDE共轭物（DS 2.4）在脾脏、肝脏和肾脏中更有效地递送pDNA。α-CDE共轭物（G3，DS 2.4）有望作为一种非病毒载体在体外和体内使用，这些数据可能有助于设计与其他非病毒载体的α-CyD共轭物。
