Design, syntheses and in vitro gene delivery efficacies of novel mono-, di- and trilysinated cationic lipids: A structure-activity investigation
Design, syntheses and in vitro gene delivery efficacies of novel mono-, di- and trilysinated cationic lipids: A structure-activity investigation
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DOI：
10.1021/jm030541
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发表时间：
2004-04-08
影响因子：
7.3
通讯作者：
Chaudhuri, A
中科院分区：
文献类型：
--
作者：
Karmali, PP;Kumar, VV;Chaudhuri, A
本发明描述了在CHO、COS-1和HepG 2细胞中的结构-活性研究，包括设计、合成和评价含有单-、二-和三赖氨酸头基的新型系列阳离子两亲物（1-10）的相对体外基因递送功效。观察到几个有趣的和相当意外的转染概况。一般而言，与（a）具有较长烃尾的单赖氨酸类似物（脂质2-4）、（B）二赖氨酸（脂质5-7）和三赖氨酸头基类似物（脂质8-10）和（c）在其极性区域具有多个正电荷的市售LipofectAmine相比，与DOPE组合使用的具有肉豆蔻基尾的脂质1显示出上级的转染性质。作为这些新的赖氨酸化阳离子脂质的相对DNA压缩功效的初步估计，使用动态激光光散射技术测量代表性脂质复合物的流体动力学直径。我们的lipoplex大小的数据是一致的概念，即共价接枝的阳离子脂质的头基区域中的带正电荷的官能团的数量不断增加，不一定会导致更紧凑的lipoplex。凝胶阻滞和DNA酶I敏感性试验表明，所有新的单，二，和三赖氨酸化的阳离子脂质类似的脂质/DNA结合的相互作用。基于MTT测定法的细胞活力结果清楚地表明，三赖氨酸类似物（8-10）与它们的单赖氨酸（1-4）和二赖氨酸（5-7）对应物相比的总体较低的转染性质不太可能源于不同的毒性相关效应。总之，本研究结果支持这样的观点，即在通过在头基区域共价接枝多个赖氨酸官能团来确保阳离子脂质的增强的基因递送功效时需要谨慎。
Structure-activity investigation including design, syntheses, and evaluation of relative in vitro gene delivery efficacies of a novel series of cationic amphiphiles (1-10) containing mono-, di-, and trilysine headgroups are described in CHO, COS-1, and HepG2 cells. Several interesting and rather unexpected transfection profiles were observed. In general, lipid 1 with the myristyl tail used in combination with DOPE as colipid exhibited superior transfection properties compared to (a) the monolysinated analogues with longer hydrocarbon tails (lipids 2-4), (b) the dilysine (lipids 5-7) and the trilysine headgroup analogues (lipids 8-10), and (c) commercially available LipofectAmine with multiple positive charges in its polar region. As a preliminary estimate of the relative DNA-compacting efficacies of these new lysinated cationic lipids, the hydrodynamic diameters of representative lipoplexes were measured using dynamic laser light scattering technique. Our lipoplex size data are consistent with the notion that covalent grafting of an increasing number of positively charged functional groups in the headgroup region of cationic lipids need not necessarily result in more compacted lipoplexes. Both gel retardation and DNase I sensitivity assays indicated similar lipid/DNA binding interactions for all the novel mono-, di-, and trilysinated cationic lipids. MTT-assay-based cell viability results clearly demonstrate that the overall lower transfection properties of trilysine analogues (8-10) compared to their mono- (1-4) and dilysinated (5-7) counterparts are unlikely to originate from differential toxicity related effects. Taken together, the present findings support the notion that caution needs to be exercised in ensuring enhanced gene delivery efficacies of cationic lipids through covalent grafting of multiple lysine functionalities in the headgroup region.