A biodegradable poly(amido amine) based on the antimicrobial polymer polyhexamethylene biguanide for efficient and safe gene delivery
A biodegradable poly(amido amine) based on the antimicrobial polymer polyhexamethylene biguanide for efficient and safe gene delivery
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基于抗菌聚合物聚六亚甲基双胍的可生物降解聚（酰胺基胺），用于高效、安全的基因传递
DOI：
10.1016/j.colsurfb.2019.110355
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
Pingtian Ding
中科院分区：
文献类型：
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作者：
Haonan Xing;Lin Cheng;Mei Lu;Hui Liu;Lang Lang;Tianzhi Yang;Xiaoyun Zhao;Hui Xu;Li Yang;Pingtian Ding
受抗菌聚合物优异的膜亲和性的启发，我们合成了一种新型的具有侧链的可生物降解的聚（氨基胺）聚合物，其模拟了广泛使用的用于基因递送的生物杀灭剂聚双胍（PHMB）。以N，N′-胱胺双丙烯酰胺（CBA）和N-叔丁氧羰基-1，6-己二胺（Boc-DAH）为原料，通过Michael加成聚合反应制备了CBA/DAH高分子骨架，然后进行N-叔丁氧羰基脱保护。然后通过与双氰胺的加成反应将暴露的伯氨基部分（约75%）转化为双胍，得到最终产物CBA-DAH-biguanide（CBA-DAH-BG）。聚合物CBA-DAH-BG能够将质粒DNA（pDNA）缩合成纳米尺寸（<200 nm）、带正电荷（>35 mV）的聚合复合物，其对肝素和DNA酶I具有良好的抗性。在二硫苏糖醇（DTT）的存在下，CBA-DAH-BG具有快速的DNA释放能力，通过二硫键的断裂使其具有生物降解性，有助于DNA解包，降低细胞毒性。CBA-DAH-BG/pDNA聚合复合物的特征在于在两种细胞系（即，NIH/3 T3和U87 MG），与均不含双胍基团的25 kDa聚乙烯亚胺（PEI）和中间产物CBA-DAH相比。值得注意的是，网格蛋白介导的内吞作用和脂筏在复合物的内化中发挥了重要作用。总之，本文描述的这种策略可以代表用于设计具有高转染效率和生物相容性的更先进的非病毒基因载体的创新途径。
Inspired by the excellent membrane affinity of antimicrobial polymers, we synthesized a novel biodegradable poly(amino amine) polymer with pendent side chains that mimic the widely used biocide polyhexamethylene biguanide (PHMB) for gene delivery. Michael addition polymerization was utilized to form the polymer scaffold betweenN,N′-cystaminebisacrylamide (CBA) andN-Boc-1,6-diaminohexane (Boc-DAH) followed by N-Boc deprotection. Then the exposed primary amino groups were partly (about 75%) transformed into biguanide by an addition reaction with dicyandiamide to obtain the final product CBA-DAH-biguanide (CBA-DAH-BG). The polymer CBA-DAH-BG was able to condense plasmid DNA (pDNA) into nano-sized (<200 nm), positively-charged (>35 mV) polyplexes that were well resistant to heparin and DNase I. Rapid DNA release was observed in the presence of dithiothreitol (DTT), indicating that CBA-DAH-BG was equipped with biodegradability by the cleavage of disulfide bonds, which was helpful for unpacking DNA and decreasing cytotoxicity. CBA-DAH-BG/pDNA polyplexes were characterized by efficient cellular uptake efficacy, extremely low cytotoxicity, and high transfection efficiency in two cell lines (i.e., NIH/3T3 and U87 MG), compared to 25 kDa polyethyleneimine (PEI) and the intermediate product CBA-DAH that were both devoid of biguanide groups. Of note, clathrin-mediated endocytosis and lipid rafts played an important role in the internalization of the polyplexes. Taken together, this strategy described herein may represent an innovative avenue for the design of more advanced nonviral gene vectors with high transfection efficiency and biocompatibility.