Tanaka Ko, Kanazawa Takanori, Ogawa Takaya, Suda Yumiko, Takashima Yuuki, Fukuda Tsunehiko, Okada Hiroaki
Laboratory of Pharmaceutics and Drug Delivery, Department of Pharmaceutical Science, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Hachioji, Tokyo, Japan.
Chem Pharm Bull (Tokyo). 2011;59(2):202-7. doi: 10.1248/cpb.59.202.
We have engineered a novel, non-viral, multifunctional gene vector (STR-CH(2)R(4)H(2)C) that contained stearoyl (STR) and a block peptide consisting of Cys (C), His (H), and Arg (R). STR-CH(2)R(4)H(2)C can form a stable nano-complex with plasmid DNA (pDNA) based on electronic interactions and disulfide cross linkages. In this study, we evaluated the efficacy of STR-CH(2)R(4)H(2)C as a gene vector. We first determined the optimal weight ratio for STR-CH(2)R(4)H(2)C/pDNA complexes. The complexes with a weight ratio of 50 showed the highest transfection efficacy. We also examined the transfection efficacy of STR-CH(2)R(4)H(2)C/pDNA complexes with or without serum and compared STR-CH(2)R(4)H(2)C/pDNA transfection efficacy with that of Lipofectamine. Even in the presence of serum, STR-CH(2)R(4)H(2)C showed higher transfection efficacy than did Lipofectamine. In addition, we determined the mechanism of transfection of the STR-CH(2)R(4)H(2)C/pDNA complexes using various cellular uptake inhibitors and evaluated its endosomal escape ability using chloroquine. Macropinocytosis was main cellular uptake pathway of STR-CH(2)R(4)H(2)C/pDNA complexes. Our results suggested that STR-CH(2)R(4)H(2)C is a promising gene delivery system.
我们构建了一种新型的、非病毒的多功能基因载体（STR-CH₂R₄H₂C），它含有硬脂酰基（STR）以及由半胱氨酸（C）、组氨酸（H）和精氨酸（R）组成的嵌段肽。基于静电相互作用和二硫键交联，STR-CH₂R₄H₂C能与质粒DNA（pDNA）形成稳定的纳米复合物。在本研究中，我们评估了STR-CH₂R₄H₂C作为基因载体的效果。我们首先确定了STR-CH₂R₄H₂C/pDNA复合物的最佳重量比。重量比为50的复合物显示出最高的转染效率。我们还检测了有血清和无血清条件下STR-CH₂R₄H₂C/pDNA复合物的转染效率，并将STR-CH₂R₄H₂C/pDNA的转染效率与脂质体转染试剂进行了比较。即使在有血清存在的情况下，STR-CH₂R₄H₂C也比脂质体转染试剂表现出更高的转染效率。此外，我们使用各种细胞摄取抑制剂确定了STR-CH₂R₄H₂C/pDNA复合物的转染机制，并使用氯喹评估了其从内涵体逃逸的能力。巨胞饮作用是STR-CH₂R₄H₂C/pDNA复合物主要的细胞摄取途径。我们的结果表明，STR-CH₂R₄H₂C是一种有前景的基因递送系统。
