Biotechnology Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, P.O. Box: 91775-1365, Mashhad, Iran.
Department of Medicinal Chemistry, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.
AAPS PharmSciTech. 2019 Feb 12;20(3):111. doi: 10.1208/s12249-018-1235-5.
Development of efficient non-viral carriers is one of the major challenges of gene delivery. In the current study, we designed, synthesized, and evaluated the in vitro gene delivery efficiency of novel amphiphilic constructs composed of cholesterol and low molecular weight protamine (LMWP: VSRRRRRRGGRRRR) peptide. Vectors having both hydrophobic and hydrophilic moieties were evaluated in terms of particle size and charge, DNA condensation ability, cytotoxicity, and gene transfection efficiency. The prepared vectors spontaneity self-assembled into the liposome-like particles with a high local positive density. The nano-vehicle A (H5-LMWP-Cholestrol) and nano-vehicle B (LMWP-Cholesterol) could form micelles at concentrations above 50 μg/mL and 65 μg/mL, respectively. The gel retardation assay showed that nano-vehicles A and B could condense pDNA more efficiently than the corresponding unconjugated peptides. The mean of size and zeta potential of complexed nano-vehicle A at N/P ratios of 5, 15, and 30 were 151 nm and 23 mv, and those of nano-vehicle B were 224 nm and 19 mv, respectively. In terms of transfection efficiency, the designed nano-vehicles showed almost two-fold higher gene expression level compared to PEI 25 kDa at optimal N/P ratios, and also exhibited negligible cytotoxicity on a model cancer cell, Neuro 2a. The findings of the present study revealed that these cationic micelles can be promising candidates as non-viral gene delivery vehicles.
开发高效的非病毒载体是基因传递的主要挑战之一。在本研究中，我们设计、合成并评估了由胆固醇和低分子量鱼精蛋白（LMWP：VSRRRRRRGGRRRR 肽）组成的新型两亲性构建体的体外基因传递效率。根据粒径和电荷、DNA 凝聚能力、细胞毒性和基因转染效率评估了具有亲水性和疏水性部分的载体。制备的载体自发自组装成具有局部正密度的类脂质体颗粒。纳米载体 A（H5-LMWP-胆固醇）和纳米载体 B（LMWP-胆固醇）可以在浓度高于 50μg/mL 和 65μg/mL 时分别形成胶束。凝胶阻滞实验表明，纳米载体 A 和 B 比相应的未缀合肽更有效地浓缩 pDNA。在 N/P 比为 5、15 和 30 时，复合纳米载体 A 的平均粒径和 Zeta 电位分别为 151nm 和 23mv，复合纳米载体 B 的平均粒径和 Zeta 电位分别为 224nm 和 19mv。就转染效率而言，与 25kDa 的 PEI 相比，设计的纳米载体在最佳 N/P 比下表现出近两倍的更高基因表达水平，并且在模型癌细胞 Neuro 2a 上也表现出可忽略的细胞毒性。本研究的结果表明，这些阳离子胶束可以作为非病毒基因传递载体的有前途的候选物。
