血管内皮细胞靶向性Tyr-RGD-PEG-PEI纳米基因载体的合成及其生物特性的初步研究  

Vascular endothelial cells targeted Tyr-RGD-PEG-PEI nano-drug synthesis and its biological activity

作　　者：高勇[1] 何胜利[2] 钟高仁[3] 蔡洪培[4] 李琦[1] 

机构地区：[1]同济大学附属东方医院肿瘤科,上海200120 [2]复旦大学附属上海市第五人民医院肿瘤科,上海200240 [3]复旦大学放射医学研究所,上海200032 [4]第二军医大学附属长征医院消化科,上海200003

出　　处：《药学学报》2009年第9期1034-1039,共6页Acta Pharmaceutica Sinica

基　　金：上海市科学技术委员会纳米专项基金资助项目(05nm05007)

摘　　要：本文设计和合成了具有血管内皮细胞靶向性的环五肽RGD(cyclo-[Arg-Gly-Asp-D-Tyr-Lys],Tyr-RGD)修饰的纳米基因载体聚乙二醇-聚乙烯亚胺(PEG-PEI),并研究其纳米特性、载药特性及其体内外的肿瘤靶向特性。采用化学方法合成所需要的纳米载体,鉴定该纳米载体的纳米特性及载药特性,并采用体外血管内皮细胞竞争结合实验研究其靶向性,最后将荧光标记的纳米载体注入荷瘤小鼠体内,观察其肿瘤靶向性。Tyr-RGD-PEG-PEI纳米载体采用分步合成方法,所获得的载体粒径约为145nm,其对小分子干扰核糖核酸(siRNA)的包封率好,血浆内孵育30min,仅约3%的siRNA游离出来,结构稳定。薄层层析法分析鉴定其为单一斑点(Rf0.65)。受体竞争实验表明该纳米载体可有效地与多肽精氨酸-甘氨酸-天冬氨酸(RGD)竞争结合内皮细胞膜上的受体;荷瘤小鼠实验表明,载有荧光标记siRNA的Tyr-RGD-PEG-PEI纳米药物注射入小鼠体内24h后,主要分布在肿瘤组织内,而无Tyr-RGD靶向的纳米药物则除了瘤体内,还分布在肝脏、肺等脏器。合成的Tyr-RGD靶向纳米载体Tyr-RGD-PEG-PEI具有较好的纳米特性和高效的基因药物包封率,荷瘤裸鼠研究表明其肿瘤的靶向性明显优于无Tyr-RGD靶向的纳米药物。The study is designed to synthesize nano-carrier Tyr-RGD （cyclo-[Arg-Gly-Asp-d-Tyr-Lys]） and poly（ethylene glycol） modified polyethylenimine （Tyr-RGD-PEG-PEI） targeting vascular endothelial cells, then analyze its nanoparticle properties and the characteristics of drug carrying and targeting properties in vivo I in vitro tumor. The nano-carrier Tyr-RGD-PEG-PEI was synthesized with the method of chemical synthesis and the properties of this nanoparticle and drug carrying characteristics were identified. Its effect of targeting vascular endothelial cells in vitro was studied with the method of competitive binding assay. The fluorescent labeled nano-drug was injected into tumor-bearing nude mice to observe its tumor-targeting. The mean size of nanocarrier Tyr-RGD-PEG-PE was about 145 nm, good in encapsulation efficiency of siRNA. After incubation in plasma for half an hour, only about 3 percent of siRNA out. It was confirmed that it was a single spot with TLC analysis, the Rf value was 0.65. Receptor competition experiments showed that the nano could effectively compete with RGD in binding the receptors on endothelial cells. Tumor-bearing nude mice experiments showed that when containing a fluorescent-labeled siRNA of Tyr-RGD-PEG-PEI nano-drug was injected into mice, after 24 hours this nano-drug mainly distributed within the tumor tissue. However, nano-drug without Tyr-RGD appeared in tumor tissue as well as other organs such as livers, lungs, etc. The Tyr-RGD-targeted gene vector Tyr-RGD-PEG-PEI synthesized in this study has good nanoparticle properties and high efficiency of gene-drug encapsulation. Study of nude mice shows that the ability of its tumor-targeting is significantly better than nano-drug without Tyr-RGD.

关 键 词：整合素 纳米 肿瘤 内皮细胞 小分子干扰核糖核酸 

分 类 号：R943[医药卫生—药剂学]