Diquaternary ammonium compounds as transfection agents.
Diquaternary ammonium compounds as transfection agents.
复制标题
双季铵化合物作为转染剂。
DOI：
10.1021/bc000099z
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发表时间：
2001
影响因子：
4.7
通讯作者：
MacDonald,RC
中科院分区：
文献类型：
--
作者：
Rosenzweig,HS;Rakhmanova,VA;MacDonald,RC
双季铵盐是介导哺乳动物细胞系 DNA 转染的新型试剂。N,N'-二油基-N,N,N',N'-四甲基-1,2-乙二胺 (TmedEce),N,N'-二油基-N,N,N',N'-四甲基-1,3-丙二胺(PropEce)、N,N'-二油基-N,N,N',N'-四甲基-1,6-己二胺 (HexEce) 及其相应的 N,N'-二十六烷基饱和类似物（TmedAce、PropAce 和 HexAce）均已合成并表征。它们是通过相应的四甲基二胺与 2.2 M 当量的长链烷基卤（饱和或不饱和）的双 Menshutkin 反应制备的。反应在无水乙腈中进行约。在 60 °C 下反应 3 天，对于大多数衍生物来说，二季铵卤化物的收率良好，甚至接近定量。使用这些新试剂在体外实现了与市售试剂（例如 Lipofectin、Lipofectace、Lipofectamine 和 O-乙基二油酰磷脂酰胆碱三氟甲磺酸盐）相当的 DNA 转染。不需要使用Colipid来有效转染，但血清确实显着抑制转染。饱和和不饱和衍生物的水合行为不同。饱和衍生物似乎在水合时保留了层状晶体阵列结构，而不饱和衍生物形成胶束和/或脂质体，具体取决于离子强度： HexEce 在水和盐水中都是胶束； PropEce 在水中呈胶束状，但在盐水中呈层状； TmedEce 两者都是层状的。尽管水合模式不同，所有这些不饱和衍生物均与 DNA 形成高效的转染复合物。改变四级位点之间的距离对饱和衍生物影响转染功效的顺序为 HexAce > TmedAce = PropAce，对于不饱和衍生物，影响转染功效的顺序为 PropEce = HexEce > TmedEce，差异较小。
Diquaternary ammonium salts constitute a new class of reagent for mediating transfection of DNA in mammalian cell lines.N,N‘-dioleyl-N,N,N‘,N‘-tetramethyl-1,2-ethanediamine (TmedEce),N,N‘-dioleyl-N,N,N‘,N‘-tetramethyl-1,3-propanediamine (PropEce),N,N‘-dioleyl-N,N,N‘,N‘-tetramethyl-1,6-hexanediamine (HexEce), and their correspondingN,N‘-dicetyl saturated analogues (TmedAce, PropAce and HexAce) have all been synthesized and characterized. They were prepared via a bis-Menshutkin reaction of the corresponding tetramethyldiamine with 2.2 M equiv of a long-chain alkyl halide (saturated or unsaturated). The reaction was run in anhydrous acetonitrile for ca. 3 days at 60 °C, which produced the diquaternary ammonium halides in good to nearly quantitative yields for most derivatives. DNA transfection comparable to commercially available reagents such as Lipofectin, Lipofectace, Lipofectamine, andO-ethyldioleoylphosphatidylcholinium triflate has been achieved in vitro with these new reagents. There was no need to use a colipid for effective transfection, but serum did significantly inhibit transfection. The saturated and the unsaturated derivatives differed with respect to hydration behavior. The saturated derivatives appeared to retain a lamellar-type crystalline array structure upon hydration, whereas the unsaturated versions formed micelles and/or liposomes, depending on the ionic strength: HexEce was micellar in both water and saline; PropEce was micellar in water but lamellar in saline; and TmedEce was lamellar in both. Despite these different hydration patterns, all of these unsaturated derivatives formed productive transfection complexes with DNA. Varying the distance between the quaternary sites affected transfection efficacy in the order HexAce > TmedAce = PropAce for the saturated derivatives and in the order PropEce = HexEce > TmedEce, with a smaller spread, for the unsaturated derivatives.