具有还原敏感可断裂的疏水尾链的新型阳离子脂质用于siRNA递送的研究

doi:10.5246/jcps.2018.06.039

中国药学(英文版) ›› 2018, Vol. 27 ›› Issue (6): 383-396.DOI: 10.5246/jcps.2018.06.039

具有还原敏感可断裂的疏水尾链的新型阳离子脂质用于siRNA递送的研究

闫仪¹, 崔仕贺¹, 孙晶¹, 李飘飘¹, 张海涛^1,2, 王坚成^1*

1. 北京大学医学部药学院分子药剂学与新释药系统北京市重点实验室, 北京 100191
2. 中南大学药学院, 湖南长沙 410013

收稿日期:2018-05-06 修回日期:2018-05-28 出版日期:2018-06-30 发布日期:2018-06-03
通讯作者: Tel.: +86-010-82805932, E-mail: wang-jc@bjmu.edu.cn
基金资助:
National Natural Science Foundation of China (Grant No. 81473158, 81690264 and 81773650), the New Drug R&D program of China (Grant No. 2018ZX09721003-004) and the Opening Project of Key Laboratory of Drug Targeting and Drug Delivery System, Ministry of Education (Sichuan University).

Novel cationic lipid with reduction-responsive cleavable hydrophobic tail for siRNA delivery

Yi Yan¹, Shihe Cui¹, Jing Sun¹, Piaopiao Li¹, Haitao Zhang^1,2, Jiancheng Wang^1*

1. Beijing Key Laboratory of Molecular Pharmaceutics and New Drug Delivery Systems, School of Pharmaceutical Sciences, Peking University Health Science Center, Beijing 100191, China
2. School of Pharmaceutical Sciences, Central South University, Changsha 410013, Hunan, China

Received:2018-05-06 Revised:2018-05-28 Online:2018-06-30 Published:2018-06-03
Contact: Tel.: +86-010-82805932, E-mail: wang-jc@bjmu.edu.cn
Supported by:
National Natural Science Foundation of China (Grant No. 81473158, 81690264 and 81773650), the New Drug R&D program of China (Grant No. 2018ZX09721003-004) and the Opening Project of Key Laboratory of Drug Targeting and Drug Delivery System, Ministry of Education (Sichuan University).

摘要/Abstract

摘要：

为了获得更高的转染效率和更低的毒性, 本研究合成了一种新型的人字形阳离子脂质(2ssHLL), 它由亲水性天冬氨酸和两条还原敏感可断裂的疏水油酸尾链组成。通过siRNA和基于2ssHLL的脂质体间的静电相互作用, 成功制备了粒径约150 nm的均匀的球形阳离子纳米复合物。从评估结果可以看出, 该纳米复合物在HepG2细胞试验中表现出较好的细胞摄取和较低的细胞毒性。RT-PCR分析结果表明, 2ssHLL/siEGFR纳米复合物可表现出与Lipofectamine2000相类似的对靶mRNA显著的下调效应。这些增强的siRNA基因沉默效率可能归因于还原响应性断裂所诱导的疏水尾链的分离。在还原环境中观察到的纳米复合物的粒径和siRNA释放的变化也证实了上述的机制。由此, 我们认为氧化还原响应型2ssHLL将有可能成为siRNA递送的潜在纳米载体。

关键词: 小干扰RNA的递送, 二硫键, 还原敏感, 纳米复合物, 断裂

Abstract:

To achieve a higher transfection efficiency and lower toxicity, a novel herringbone-like cationic lipid (2ssHLL) composed of hydrophilic aspartic acid linked with two reduction-responsive cleavable hydrophobic oleic acid tails was synthesized and assessed in this study. In our results, the cationic nanoplexes with a uniform spherical shape and a particle size of ~150 nm were successfully prepared by the electrostatic interaction between siRNAs and 2ssHLL-based liposomes. From the results evaluated in HepG2 cells, it was shown that the nanoplexes exhibited high cellular uptake of siRNA with a low cytotoxicity. Moreover, the significant down-regulation effects of 2ssHLL/siEGFR nanoplexes on target mRNA were displayed by RT-PCR analysis, which were similar to those of Lipofectamine2000. It suggested that the enhanced siRNA gene silencing efficiency was probably attributed to the detachment of hydrophobic tail chains induced by reduction-responsive cleavage. This mechanism was also confirmed by the changes of size distribution and siRNA release of nanoplexes in the reductive environment and DTT-absence condition. Overall, we believed that the redox-active herringbone-like 2ssHLL would be a potential nanocarrier towards siRNA delivery.

Key words: siRNA delivery, Disulfide bond, Reduction-responsive, Nanoplexes, Cleavage

中图分类号:

R943

Supporting:

闫仪, 崔仕贺, 孙晶, 李飘飘, 张海涛, 王坚成. 具有还原敏感可断裂的疏水尾链的新型阳离子脂质用于siRNA递送的研究[J]. 中国药学(英文版), 2018, 27(6): 383-396.

Yi Yan, Shihe Cui, Jing Sun, Piaopiao Li, Haitao Zhang, Jiancheng Wang. Novel cationic lipid with reduction-responsive cleavable hydrophobic tail for siRNA delivery[J]. Journal of Chinese Pharmaceutical Sciences, 2018, 27(6): 383-396.

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具有还原敏感可断裂的疏水尾链的新型阳离子脂质用于siRNA递送的研究

Novel cationic lipid with reduction-responsive cleavable hydrophobic tail for siRNA delivery

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