O-ethylphosphatidylcholine:: A metabolizable cationic phospholipid which is a serum-compatible DNA transfection agent
O-ethylphosphatidylcholine:: A metabolizable cationic phospholipid which is a serum-compatible DNA transfection agent
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DOI：
10.1021/js990006q
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发表时间：
1999-09-01
影响因子：
3.8
通讯作者：
Lahiri, MK
中科院分区：
文献类型：
--
作者：
MacDonald, RC;Rakhmanova, VA;Lahiri, MK
以磷脂酰胆碱为原料，与三氟甲烷磺酸乙酯一步反应制得1,2-二甘油酯-sn-甘油-3-乙基磷酸胆碱。这和相关的o -烷基磷脂酰胆碱构成了生物脂结构的第一个化学稳定的三酯，也是第一个完全由酯键连接的生物代谢物组成的磷脂的阳离子衍生物。阳离子磷脂与质粒DNA复合物高效转染细胞。转染效率最高的复合物中，正电荷比负电荷多几个百分点。通过在10%血清中连续培养，获得了普通细胞系转染的适度刺激。磷脂复合物在37℃几乎纯净的血清中孵育至少2小时，对转染效率没有有害影响。脂质毒性低；BHK细胞耐受剂量为2mg /2 × 10(6)个细胞，浓度为1mg /mL。脂质是可生物降解的；它在体外被磷脂酶A(2)水解，在培养细胞中代谢，半衰期为几天。阳离子磷脂的合成路线非常适合于制备具有各种不同性质的定制衍生物。
1,2-Dioleoyl-sn-glycero-3-ethylphosphocholine was prepared in a one-step reaction from phosphatidylcholine by reaction with ethyl trifluoromethanesulfonate. This and related O-alkyl phosphatidylcholines constitute the first chemically stable triesters of biological lipid structures and the first cationic derivatives of phospholipids consisting entirely of biological metabolites linked with ester bonds. The complex of cationic phospholipid and plasmid DNA transfected cells with high efficiency. Maximum efficiency of transfection was obtained with complexes in which the positive charge was a few percent in excess over the negative charge. Modest stimulation of transfection of common cell lines was obtained by continuous culture in the presence of 10% serum. Incubation of the phospholipid complex for at least 2 h at 37 degrees C in nearly pure serum had no deleterious effects on transfection efficiency. The lipid has low toxicity; BHK cells tolerated amounts of 2 mg/2 x 10(6) cells at concentrations of 1 mg/mL. The lipid is biodegradable; it was hydrolyzed by phospholipase A(2) in vitro and was metabolized with a half-life of a few days in cells in culture. The synthetic route to cationic phospholipids is well suited to the preparation of derivatives that are tailor-made to have a wide variety of different properties.