Structural and formulation factors influencing pyridinium lipid-based gene transfer.
Structural and formulation factors influencing pyridinium lipid-based gene transfer.
复制标题
影响吡啶鎓脂质基因转移的结构和配方因素。
DOI：
10.1021/bc8004039
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发表时间：
2008-12
影响因子：
4.7
通讯作者：
Mahato RI
中科院分区：
文献类型：
--
作者：
Zhu L;Lu Y;Miller DD;Mahato RI
合成了一系列含杂环和氮原子的吡啶脂质，以确定基因递送的构效关系。以异亚丙基丙酮为原料，通过单酰化反应合成了氯铝酸吡喃，并将其转化为六氟磷酸吡喃，以其为关键中间体与不同伯胺反应，得到六氟磷酸羟乙基吡啶和六氟磷酸氨乙基吡啶。用不同类型的脂肪酰氯酰化这些吡啶鎓盐得到最终的吡啶鎓脂质，将其与共脂质如L-α-二油酰基磷脂酰乙醇胺（DOPE）和胆固醇（Chol）混合以通过超声处理制备阳离子脂质体。将这些脂质体与编码增强型绿色荧光蛋白（pCMS-EGFP）或荧光素酶（pDNA 3-Luc）的质粒DNA混合，并转染中国人卵巢（CHO）细胞。疏水锚链长度、锚链类型、双键构型、连接体类型、共脂质类型、阳离子脂质与共脂质的摩尔比、电荷比（N/P）和细胞类型等因素对转染效率和细胞毒性有显著影响。具有酰胺接头的吡啶鎓脂质与其酯对应物相比显示出显著更高的转染效率。当使用DOPE或胆固醇作为共脂质以制备阳离子脂质体用于与质粒DNA以3：1（+/-）电荷比形成复合物时，以1：1摩尔比的吡啶鎓脂质和共脂质制备的脂质体显示出更高的转染效率。基于疏水锚链长度为16的吡啶鎓脂质体显示出更高的转染效率和更低的细胞毒性。在相同的脂肪酸链长度下，脂肪酸链中双键为反式构型的吡啶脂质比顺式构型的吡啶脂质具有更高的转染效率。在血清存在下，C16：0和Lipofectamine显著降低了它们的转染效率，在30%及更高的血清浓度下完全丧失，而C16：1反式异构体在这些条件下仍然具有高的转染效率。总之，吡啶脂质体具有较高的转染效率，有潜力作为体外和体内转染试剂。
A series of pyridinium lipids containing a heterocyclic ring and a nitrogen atom were synthesized to determine the structure-activity relationship for gene delivery. Pyrylium chloroaluminate was synthesized by monoacylation of mesityl oxide and converted into pyrylium hexafluorophosphate, which was used as the key intermediate for reaction with different primary amines, to yield hydroxyethylpyridinium hexafluorophosphate and aminoethylpyridinium hexafluorophosphate. Acylation of these pyridinium salts with different types of fatty acid chlorides afforded the final pyridinium lipids, which were mixed with a co-lipid, such as L-alpha-dioleoylphosphatidylethanolamine (DOPE) and cholesterol (Chol) to prepare cationic liposomes by sonication. These liposomes were mixed with plasmid DNA encoding enhanced green fluorescent protein (pCMS-EGFP) or luciferase (pDNA3-Luc) and transfected into Chinese Hamster Ovary (CHO) cells. Several factors including hydrophobic anchor chain length, anchor chain type, configuration of double bond, linker type, co-lipid type, cationic lipid: co-lipid molar ratio, charge ratio (N/P), and cell type had significant influence on transfection efficiency and cytotoxicity. Pyridinium lipids with amide linker showed significantly higher transfection efficiency compared to their ester counterparts. Liposomes prepared at 1:1 molar ratio of pyridinium lipid and co-lipid showed higher transfection efficiency when either DOPE or cholesterol was used as a co-lipid to prepare cationic liposomes for complex formation with plasmid DNA at 3:1(+/−) charge ratio. Pyridinium liposomes based on hydrophobic anchor chain length of 16 showed higher transfection efficiency and lower cytotoxicity. The pyridinium lipid with trans-configuration of the double bond in fatty acid chain showed higher transfection efficiency than its counterpart with cis-configuration at the same fatty acid chain length. In the presence of serum, C16:0 and Lipofectamine significantly decreased their transfection efficiencies, which were completely lost at the serum concentration of 30% and higher, while C16:1 trans-isomer still had high transfection efficiency under these conditions. In conclusion, pyridinium lipids showed high transfection efficiency and have the potential to be used as transfection reagents in vitro and in vivo.
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影响因子：
7.3
作者：
Haldar, J;Kondaiah, P;Bhattacharya, S
通讯作者：
Bhattacharya, S
影响因子：
7.3
作者：
Ilies, MA;Seitz, WA;Balaban, AT
通讯作者：
Balaban, AT
影响因子：
2.9
作者：
Miller, CR;Bondurant, B;O'Brien, DF
通讯作者：
O'Brien, DF
DOI：
10.1016/0005-2736(90)90017-i
发表时间：
1990-03-30
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA
影响因子：
--
作者：
LEVENTIS, R;SILVIUS, JR
通讯作者：
SILVIUS, JR
DOI：
10.1016/0005-2736(92)90316-e
发表时间：
1992-11-09
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA
影响因子：
--
作者：
FARHOOD, H;BOTTEGA, R;HUANG, L
通讯作者：
HUANG, L