Kurosaki Tomoaki, Kitahara Takashi, Teshima Mugen, Nishida Koyo, Nakamura Junzo, Nakashima Mikiro, To Hideto, Hukuchi Hiromitsu, Hamamoto Tomoyuki, Sasaki Hitoshi
Department of Hospital Pharmacy, University Hospital of Medicine and Dentistry, 1-7-1 Sakamoto, Nagasaki 852-8501, Japan.
J Pharm Pharm Sci. 2008;11(4):56-67. doi: 10.18433/j31s3b.
In gene delivery, a fusogenic lipid such as dioleyl phosphatidylethanolamine (DOPE) which is a component of cationic liposomal vector is important factor for effective transfection efficiency. We investigated the effect of penetration enhancers as alternative helper-lipids to DOPE.
Transdermal penetraion enhancers such as N-lauroylsarcosine (LS), (R)-(+)-limonene (LM), vitamin E (VE), and phosphatidyl choline from eggs (EggPC) were used in this experiments as helper-lipids with N-[1-(2, 3-dioleyloxy) propyl]-N, N, N-trimethlylammonium chloride (DOTMA) and cholesterol (CHOL). We examined in vitro transfection efficiency, cytotoxicity, hematotoxicity, and in vivo transfection efficiency of plasmid DNA/cationic liposomes complexes.
In transfection experiments in vitro, the cationic lipoplexes containing LS had highest transfection efficiency among the other lipoplexes independently of FBS. Furthermore, the lipoplexes containing LS had lowest cell toxicity among the other lipoplexes in the presence of FBS. As the results of erythrocytes interaction experiment, DOTMA/LS/CHOL, DOTMA/VE/CHOL, and DOTMA/EggPC/CHOL lipoplexes showed extremely lower hematotoxicity. On the basis of these results, the in vivo transfection efficiencies of the lipoplexes were examined. The lipoplexes containing LS had the highest transfection activity among the other lipoplexes.
In conclusion, several transdermal penetration enhancers are available for alternative helper-lipids to DOPE in cationic liposomal vectors. Among them, DOTMA/LS/CHOL lipoplexes showed superior characteristics in in vitro transfection efficiency, cell toxicity, hematotoxicity, and in vivo transfection efficiency.
在基因递送中,作为阳离子脂质体载体成分的融合脂质,如二油酰磷脂酰乙醇胺(DOPE),是有效转染效率的重要因素。我们研究了渗透增强剂作为DOPE替代辅助脂质的作用。
本实验使用N-月桂酰肌氨酸(LS)、(R)-(+)-柠檬烯(LM)、维生素E(VE)和鸡蛋卵磷脂(EggPC)等透皮渗透增强剂作为辅助脂质,与N-[1-(2,3-二油酰氧基)丙基]-N,N,N-三甲基氯化铵(DOTMA)和胆固醇(CHOL)一起使用。我们检测了质粒DNA/阳离子脂质体复合物的体外转染效率、细胞毒性、血液毒性和体内转染效率。
在体外转染实验中,含LS的阳离子脂质体复合物在其他脂质体复合物中具有最高的转染效率,与胎牛血清无关。此外,在胎牛血清存在的情况下,含LS的脂质体复合物在其他脂质体复合物中细胞毒性最低。作为红细胞相互作用实验的结果,DOTMA/LS/CHOL、DOTMA/VE/CHOL和DOTMA/EggPC/CHOL脂质体复合物显示出极低的血液毒性。基于这些结果,检测了脂质体复合物的体内转染效率。含LS的脂质体复合物在其他脂质体复合物中具有最高的转染活性。
总之,几种透皮渗透增强剂可作为阳离子脂质体载体中DOPE的替代辅助脂质。其中,DOTMA/LS/CHOL脂质体复合物在体外转染效率、细胞毒性、血液毒性和体内转染效率方面表现出优异的特性。