Severance Integrative Research Institute for Cerebral and Cardiovascular Disease, Yonsei University Health System, Seoul, South Korea.
Eur J Pharm Biopharm. 2012 May;81(1):14-23. doi: 10.1016/j.ejpb.2012.01.013. Epub 2012 Jan 28.
Clinical applications of RNA interference-based therapeutics such as small interfering RNAs (siRNAs) have been limited mainly due to low intracellular delivery efficiency in vitro and in vivo. In this study, facially amphipathic deoxycholic acid (DA)-modified polyethyleneimine (PEI(1.8)) (DA-PEI(1.8)) was synthesized and used as a potent carrier system for siRNA targeted against matrix metalloproteinase-2 (MMP-2) to inhibit the migration of vascular smooth muscle cells (SMCs), which is the major pathomechanism in the development of atherosclerosis and restenosis after arterial injury. A representative facial amphipathic bile acid DA having a high membrane permeability was conjugated to the terminal amine groups of the low molecular weight PEI(1.8) via amide bonds. The DA-PEI(1.8) conjugates formed self-assembled nanoparticles with siRNA molecules in an aqueous phase and the DA-PEI(1.8)/siRNA polyplexes became stabilized and condensed as particle incubation time increased from 0 to 4h. Both cellular internalization and target gene silencing were enhanced as the DA-PEI(1.8)/siRNA polyplexes stabilized. When vascular SMCs were transfected with MMP-2 siRNA, the DA-PEI(1.8)/siRNA polyplex formulation led to a significant decrease in MMP-2 gene expression, resulting in the suppression of cell migration. These results suggest that the DA-PEI(1.8)/MMP-2 siRNA delivery system may be useful in anti-restenotic treatment for various vasculoproliferative disorders such as atherosclerosis, in-stent restenosis, and vein graft failure.
基于 RNA 干扰的治疗方法的临床应用，如小干扰 RNA（siRNA），主要受到体外和体内低细胞内递送效率的限制。在这项研究中，合成了具有面两亲性的去氧胆酸（DA）修饰的聚乙烯亚胺（PEI(1.8)）（DA-PEI(1.8)），并将其用作针对基质金属蛋白酶-2（MMP-2）的 siRNA 的有效载体系统，以抑制血管平滑肌细胞（SMC）的迁移，这是动脉损伤后动脉粥样硬化和再狭窄发展的主要病理机制。一种具有高膜通透性的代表性面两亲性胆酸 DA 通过酰胺键与低分子量 PEI(1.8)的末端胺基连接。DA-PEI(1.8) 缀合物在水相中与 siRNA 分子形成自组装纳米颗粒，并且随着粒子孵育时间从 0 增加到 4 小时，DA-PEI(1.8)/siRNA 聚集体变得稳定和凝聚。随着 DA-PEI(1.8)/siRNA 聚集体的稳定，细胞内摄取和靶基因沉默都得到增强。当血管平滑肌细胞用 MMP-2 siRNA 转染时，DA-PEI(1.8)/siRNA 聚集体配方导致 MMP-2 基因表达显著下降，从而抑制细胞迁移。这些结果表明，DA-PEI(1.8)/MMP-2 siRNA 递药系统可能对各种血管增殖性疾病（如动脉粥样硬化、支架内再狭窄和静脉移植物失败）的抗再狭窄治疗有用。
