文献：cRGD peptide-conjugated polyethylenimine-based lipid nanoparticle for intracellular delivery of siRNA in hepatocarcinoma therapy

文献链接：https://pubmed.ncbi.nlm.nih.gov/34042551/

作者：Shuang Yanga , Dandan Wangb , Xia Zhanga , Yaojun Suna and Bin Zhengc

相关产品： DSPE-PEG2000-cRGD  磷脂-聚乙二醇2000-靶向穿膜肽

https://www.ruixitech.com/product-details/dspe-peg-crgd.html

原文摘要：The effective delivery system plays an important role in the application of siRNA in the antitumor study. However, until now, researches on the delivery systems targeting hepatocarcinoma cells are still being explored. Here we designed and prepared a novel siRNA delivery system, cRGD-PSH-NP, which was based on a modified polyethyleneimine (PSH) and DSPE-PEG2000-cRGD. cRGD-PSH-NP loaded with survivin siRNA (cRGD-PSH-NP/S) was composed of egg phosphatidylcholine, cationic PSH, PEGylated lipids, survivin siRNA, and cRGD peptide as a targeting ligand. The formulations of cRGD-PSH-NP/S were optimized and characterized. In vitro investigations showed excellent gene silencing and antitumor activity compared with the unmodified nanoparticles in HepG2 cells. In vivo antitumor efficacy of cRGD-PSH-NP/S exhibited potent tumor inhibition (74.71%) in HepG2-bearing nude mice without inducing toxicity. These data suggested further research of cRGD-PSH-NP/S in hepatocarcinoma therapy.

该文献设计并制备一种新的siRNA传递系统cRGD-PSH-NP，它基于修饰的聚乙烯亚胺（PSH）和DSPE-PEG2000-cRGD。载生存素siRNA（cRGD-PSH-NP/S）的cRGD-PSH-NP由卵磷脂酰胆碱、阳离子PSH、聚乙二醇化脂质、生存素siRNA和cRGD肽作为靶向配体组成。为了增加循环时间，准确地靶向肝cancer细胞，利用DSPE-PEG2000-cRGD作为纳米颗粒的靶向配体，构成了最外层的结构。对纳米颗粒的性质进行了表征，并评价了体外和体内的转染活性。

图为：DSPE-PEG2000-cRGD的结构公式

DSPE-PEG2000-cRGD在纳米颗粒的制备及配方的优化中的应用：

采用乙醇稀释法制备脂质纳米颗粒（NP）或含survivivinsiRNA（NP/S）的脂质纳米颗粒。为了验证cRGD-PSH-NP作为siRNA的载体优于其他纳米颗粒，制备了PEINP、PSH-NP和cRGD-NP，并与生存素siRNA结合。简单地说，将含有PSH（PSH-NP）、ePC、胆固醇、PSH和DSPE-PEG2000的NP溶解在乙醇中，均匀混合得到脂质溶液。然后将脂质溶液注入PBS溶液中，以快速混合，得到不同浓度PSH的PSH-NP。对于含有siRNA的PSH-NP（PSH-NP/S），将生存素siRNA溶解在预冷的DEPC水中，加入PSH-NP溶液中混合。同时，制备PEI-NP/S作为对照组，以确定修饰后的PEI（PSH）的低有害性和高细胞摄取效率，并证明在纳米颗粒中加入PSH而不是PEI的必要性。在制备过程中，PEI-NP/S组中的PSH被PEI）取代，其他工艺与PSHNP/S的制备方法一致。对于含有cRGD（cRGD-NP）的NP，将ePC、胆固醇、PEI、DSPE-PEG2000和DSPE-PEG2000-cRGD溶解在乙醇中，快速混合注入PBS溶液中。然后将Survivin siRNA溶解在预冷的DEPC水中，加入到cRGD-NP中，然后混合，得到cRGD-NP/S。用cRGD-NP/S进行比较，说明PSH是否能提高纳米颗粒的转染效率。最后，设计并制备了代表含有cRGD和PSH的NP的cRGD-PSH-NP。

图为：cRGD-PSH-NP的结构示意图及其在siRNA传递到cancer细胞中的应用

综上所述，含有基于PEI（PSH）的阳离子聚合物和靶向配体（DSPE-PEG2000-cRGD）的纳米颗粒cRGD-PSH-NP可以传递siRNA。发现cRGD-PSH-NP的有害性较低，并且能够有效地结合siRNA。体内外实验表明，cRGD-PSH-NP/S是安全的，具有较高的抗cancer活性。认为cRGD-PSHNP可能适合和有效地用于进一步研究其他核酸化合物的传递。