Buck Jonas, Mueller Dennis, Mettal Ute, Ackermann Miriam, Grisch-Chan Hiu Man, Thöny Beat, Zumbuehl Andreas, Huwyler Jörg, Witzigmann Dominik
Division of Pharmaceutical Technology, Department of Pharmaceutical Sciences, University of Basel, Klingelbergstrasse 50/70, 4056 Basel, Switzerland.
Department of Chemistry, University of Fribourg, 1700 Fribourg, Switzerland.
ACS Omega. 2020 Sep 15;5(38):24724-24732. doi: 10.1021/acsomega.0c03303. eCollection 2020 Sep 29.
Cellular delivery of DNA vectors for the expression of therapeutic proteins is a promising approach to treat monogenic disorders or cancer. Significant efforts in a preclinical and clinical setting have been made to develop potent nonviral gene delivery systems based on lipoplexes composed of permanently cationic lipids. However, transfection efficiency and tolerability of such systems are in most cases not satisfactory. Here, we present a one-pot combinatorial method based on double-reductive amination for the synthesis of short-chain aminolipids. These lipids can be used to maximize the DNA vector delivery when combined with the cationic lipid 1,2-dioleoyl-3-trimethylammonium propane (DOTAP). We incorporated various aminolipids into such lipoplexes to complex minicircle DNA and screened these systems in a human liver-derived cell line (HuH7) for gene expression and cytotoxicity. The lead aminolipid AL-A12 showed twofold enhanced gene delivery and reduced toxicity compared to the native DOTAP:cholesterol lipoplexes. Moreover, AL-A12-containing lipoplexes enabled enhanced transgene expression in the zebrafish embryo model.
用于治疗性蛋白质表达的DNA载体的细胞递送是治疗单基因疾病或癌症的一种有前景的方法。在临床前和临床环境中，人们已经做出了巨大努力来开发基于由永久阳离子脂质组成的脂质体的高效非病毒基因递送系统。然而，在大多数情况下，此类系统的转染效率和耐受性并不令人满意。在此，我们提出了一种基于双还原胺化的一锅组合方法，用于合成短链氨基脂质。当与阳离子脂质1,2-二油酰基-3-三甲基铵丙烷（DOTAP）结合使用时，这些脂质可用于最大化DNA载体递送。我们将各种氨基脂质掺入此类脂质体中以复合小环DNA，并在人肝源细胞系（HuH7）中筛选这些系统的基因表达和细胞毒性。与天然的DOTAP:胆固醇脂质体相比，先导氨基脂质AL-A12的基因递送增强了两倍，毒性降低。此外，含AL-A12的脂质体在斑马鱼胚胎模型中能够增强转基因表达。
