“Building-block crosslinking”micelles for enhancing cellular transfection of biocompatible polycations  

“嵌段交联”胶束策略用于增强生物相容性阳离子聚合物的胞内转染效率

作　　者：Pengkai Wu Shuai Yin Tianqing Liu Dan Ding Kaikai Wang 武鹏凯;殷帅;刘天庆;丁丹;王开开(School of Pharmacy,Nantong University,Nantong,226001,China;School of Pharmacy,China Pharmaceutical University,Nanjing,210009,China;QIMR Berghofer Medical Research Institute,Brisbane,QLD,4029,Australia;State Key Laboratory of Medicinal Chemical Biology,Key Laboratory of Bioactive Materials,Ministry of Education,College of Life Sciences,Nankai University,Tianjin,300071,China)

机构地区：[1]School of Pharmacy,Nantong University,Nantong,226001,China [2]School of Pharmacy,China Pharmaceutical University,Nanjing,210009,China [3]QIMR Berghofer Medical Research Institute,Brisbane,QLD,4029,Australia [4]State Key Laboratory of Medicinal Chemical Biology,Key Laboratory of Bioactive Materials,Ministry of Education,College of Life Sciences,Nankai University,Tianjin,300071,China

出　　处：《Science China Materials》2021年第1期241-251,共11页中国科学（材料科学（英文版）

基　　金：supported by the National Natural Science Foundation of China(81903556);the Natural Science Fund for Colleges and Universities in Jiangsu Province(19KJB350004);supported by the National Health and Medical Research Council(NHMRC)Early Career Fellowship(1112258)of Australia。

摘　　要：Incorporating functional ligands and biodegradable bonds into biocompatible low-molecular-weight(LMW)polymers,such as 1.8 kDa poly(ethylenimine)(PEI1.8 k),is a common strategy to improve the properties of LMW polymers including biosafety and delivery efficacy.This study demonstrates the hypothesis that introducing different functional ligands and linked reductive disulfides in PEI 1.8k will achieve superior siRNA transfection efficiency.By incorporating PEI-X(X represents cholesterol(Ch),heptafluorobutyric anhydride(HFBA,F)and 4-carboxyphenylboronic acid(PBA))functional ligands into PEI 1.8k and subsequently crosslinking with each other via disulfide bond links,reductive-responsive PEI-X-SS-X-PEI copolymers were constructed to enhance the cellular transfection via the synergistic effect of the high affinity of Ch,F and PBA to cell membranes and the disulfide reduction triggered intracellular disassembly of micelles and subsequent siRNA release.Extraordinarily,ternary Ch-SS-F-SS-PBA micelles exhibited the strongest siRNA transfection efficiencies in in vitro cell experiments and in vivo animal experiments due to the coordination of enhanced serum stability,promoted cell uptake and endosomal escape,and cell targeting ability.This strategy of constructed multifunctional polymer here we called"building-block crosslinking"showed a simple and smart way to synthesize new materials.Also this strategy of constructing ligands-directed reduction-sensitive micelles improves the transfection efficiency of LMW PEI and provides a valuable insight to develop novel gene delivery systems.将功能性配体和生物可降解性键结合到生物相容性的低分子量(LMW)聚合物中,如1.8k聚乙烯亚胺(PEI),是提高LMW聚合物生物安全性和递送效率的常见策略.本研究证明了在PEI 1.8k中引入不同功能性配体并使用还原性二硫键将其交联可获得超高siRNA递送效率.通过将X(X代表胆固醇(Ch)、七氟丁酸酐(HFBA,F)和4-羧苯基硼酸(PBA))功能配体分别引入PEI 1.8k中得到PEI-X单体聚合物,然后将三者通过二硫键相互交联可成功构建还原响应性PEI-X-SS-X-PEI共聚物.利用Ch、F、PBA对细胞膜的高亲和力、还原性二硫键触发的细胞内胶束解体及随后siRNA释放的协同作用,PEI-X-SS-X-PEI可显著增强siRNA的细胞转染.这种三元型Ch-SS-F-SS-PBA/siRNA形成的胶束在体外细胞实验和体内动物实验中均表现出最强的siRNA转染效率,这得益于胶束具有增强的血清稳定性、提升的细胞摄取和溶酶体逃逸能力以及细胞靶向性的协同作用.这种"嵌段交联"型的多功能聚合物的构建展示了一种简单并智能的材料合成方法.同时,这种配体介导构建还原敏感型胶束的策略提高了LMW-PEI的转染效率,为开发新的基因递送系统提供了有价值的思路.

关 键 词：siRNA delivery ligand modification low-molecular-weight polymers transfection efficiency 

分 类 号：R450[医药卫生—治疗学] TQ317[医药卫生—临床医学]