基于门舒特金反应的酶触发自降解高分子载体及其在基因输送中的应用

徐昌活; 周泉; 祝鼎成; 邱娜莎; 邵世群; 刘祥瑞; 申有青

doi:10.11777/j.issn1000-3304.2017.16316

您当前的位置：

首页 >

文章列表页 >

基于门舒特金反应的酶触发自降解高分子载体及其在基因输送中的应用

研究论文 | 更新时间：2021-03-19

- 基于门舒特金反应的酶触发自降解高分子载体及其在基因输送中的应用
- Esterase-activated Self-immolative PolymericVector Based on Menschutkin ReactionforGene Delivery
- 高分子学报 2017年第6期页码：937-945
- 作者机构：
  
  生物质化工教育部重点实验室浙江大学化学工程与生物工程学院杭州 310027
- 作者简介：
  
  申有青, E-mail:shenyq@zju.edu.cn You-qing Shen, E-mail: shenyq@zju.edu.cn
- 基金信息：
  
  国家重点基础研究发展计划(2014CB931900);国家自然科学基金重大项目(51403185)
- DOI：10.11777/j.issn1000-3304.2017.16316
  中图分类号：
- 纸质出版日期：2017-6，
  
  收稿日期：2016-10-18，
  
  修回日期：2016-11-16，
扫描看全文
徐昌活, 周泉, 祝鼎成, 邱娜莎, 邵世群, 刘祥瑞, 申有青. 基于门舒特金反应的酶触发自降解高分子载体及其在基因输送中的应用[J]. 高分子学报, 2017,(6):937-945.

Chang-huo Xu, Quan Zhou, Ding-cheng Zhu, Na-sha Qiu, Shi-qun Shao, Xiang-rui Liu, You-qing Shen. Esterase-activated Self-immolative PolymericVector Based on Menschutkin ReactionforGene Delivery[J]. Acta Polymerica Sinica, 2017,(6):937-945.
徐昌活, 周泉, 祝鼎成, 邱娜莎, 邵世群, 刘祥瑞, 申有青. 基于门舒特金反应的酶触发自降解高分子载体及其在基因输送中的应用[J]. 高分子学报, 2017,(6):937-945. DOI： 10.11777/j.issn1000-3304.2017.16316.

Chang-huo Xu, Quan Zhou, Ding-cheng Zhu, Na-sha Qiu, Shi-qun Shao, Xiang-rui Liu, You-qing Shen. Esterase-activated Self-immolative PolymericVector Based on Menschutkin ReactionforGene Delivery[J]. Acta Polymerica Sinica, 2017,(6):937-945. DOI： 10.11777/j.issn1000-3304.2017.16316.

摘要

用邻位苄基溴与双胺进行门舒特金反应，合成了2种线性的季铵盐阳离子聚合物.其中，含有酚基酯键的阳离子聚合物，一旦进入细胞后，可以在细胞内的酯酶催化下快速水解，使得聚合物自降解断裂为不带电的非季铵盐小分子，从而快速释放DNA，最终达到提高转染效率的目的.通过对复合物纳米颗粒的粒径和电势测定，证明了这2种阳离子聚合物都能够有效地结合DNA形成表面带正电的复合物纳米颗粒.凝胶阻滞电泳实验表明，所合成的阳离子聚合物都能稳定地包裹DNA.而在酯酶条件下，含有酚基酯键的阳离子聚合物可以发生降解，使得纳米复合物释放出DNA.同时，含有酚基酯键的阳离子聚合物由于其独特的可降解性，相比于PEI，降低了细胞毒性.在体外细胞转染实验中，2种阳离子聚合物都有较好的转染效果.其中酯酶响应的载体在高N/P下依然表现出较高的转染效率，说明该阳离子载体能够在细胞内有效降解并释放出DNA。

Abstract

Cationic polymers have been widely explored as non-viral gene delivery vectors. The high density of cationic charges and high molecular weight of cationic polymers enable them efficiently condense DNA into nanosized polyplexes

which protect DNA from degradation and facilitate DNA cellular internalization. However

these characters also make the polyplexes hardly dissociate to release DNA

hindering its access to the DNA transcription process for gene expression. In this study

we synthesized two kinds of linear quaternized ammonium-based cationic polymers from 2

6-dibromomethylphenolic esters using Menschutkin Reaction. The esterases in cytosol could hydrolyze the phenolic ester and trigger a self-immolative chain fragmentation

leading to the quick release of the packed DNA for efficient transcription. The DLS measurements of the size and zeta potential of the resulting polyplexes showed that these cationic polymers could pack DNA molecules efficiently into positively-charged nanoparticles. Agarose-gel electrophoresis experiment revealed that anionic DNA molecules were condensed stably and compactly with cationic polymers. Furthermore

self-immolative elimination was successfully monitored by high performance liquid chromatography (HPLC)

which confirmed the esterase-responsiveness and compatibility of this well-designed cationic polymers. Gel retardation assay showed that the packed DNA could be quickly released after incubation with esterase solution

indicating that the polyplexes would become incompact and disassemble once in the cytosol. Meanwhile

compared with commercially available PEI

the polycation had a lower cytotoxicity in cancer cells

which might contribute to the fragmentation and the removal of the quaternary amine cationic charges. In A549 and Hela cancer cells

these two kinds of polyplexes achieved excellent gene transfection efficiency. But complex

was better than complex

at high N/P ratios

thanks to the activation of self-immolative elimination by intracellular esterases.

关键词

基因输送阳离子聚合物触发自降解门舒特金反应酯酶响应

Keywords

Gene deliveryCationic polymersSelf-immolativeMenschutkin ReactionEsterase-responsive

references

S C de Smedt , J Demeester , W E Hennink . . Pharm Res , 2000 . 17 ( 2 ): 113 - 126 . DOI:10.1023/A:1007548826495http://doi.org/10.1023/A:1007548826495.

T G Park , J H Jeong , S W Kim . . Adv Drug Deliv Rev , 2006 . 58 ( 4 ): 467 - 486 . DOI:10.1016/j.addr.2006.03.007http://doi.org/10.1016/j.addr.2006.03.007.

J Vacik , B S Dean , W E Zimmer , DeanDA . . Gene Ther , 1999 . 6 ( 6 ): 1006 DOI:10.1038/sj.gt.3300924http://doi.org/10.1038/sj.gt.3300924.

D V Schaffer , D A Lauffenburger . . J Biol Chem , 1998 . 273 ( 43 ): 28004 - 28009 . DOI:10.1074/jbc.273.43.28004http://doi.org/10.1074/jbc.273.43.28004.

O Boussif , F Lezoualc'h , M A Zanta , D MergnyM , D Scherman , B Demeneix , J P Behret . . Proc Natl Acad Sci USA , 1995 . 92 ( 16 ): 7297 - 7301 . DOI:10.1073/pnas.92.16.7297http://doi.org/10.1073/pnas.92.16.7297.

C Wang . . Sci Bull , 2015 . 60 ( 3 ): 403 - 404 . DOI:10.1007/s11434-014-0719-2http://doi.org/10.1007/s11434-014-0719-2.

F Zhao , H Meng , L Yan , B Wang , Y L Zhao . . Sci Bull , 2015 . 60 ( 1 ): 3 - 20 . DOI:10.1007/s11434-014-0700-0http://doi.org/10.1007/s11434-014-0700-0.

FH Meng , W E Hennink , ZY Zhong . . Biomaterials , 2009 . 30 ( 12 ): 2180 - 2198 . DOI:10.1016/j.biomaterials.2009.01.026http://doi.org/10.1016/j.biomaterials.2009.01.026.

J X Gu , W P Cheng , J G Liu , S Y Lo , D Smith , X Z Qu , Z Z Yang . . Biomacromolecules , 2007 . 9 ( 1 ): 255 - 262 . http://pubs.acs.org/doi/abs/10.1021/bm701084w.

X Liu , J J Xiang , D C Zhu , L M Jiang , Z X Zhou , J B Tang , X R Liu , Y Z Huang , Y Q Shen . . Adv Mater , 2016 . 28 ( 9 ): 1743 - 1752 . DOI:10.1002/adma.201504288http://doi.org/10.1002/adma.201504288.

S H Choi , S H Lee , T G Park . . Biomacromolecules , 2006 . 7 ( 6 ): 1864 - 1870 . DOI:10.1021/bm060182ahttp://doi.org/10.1021/bm060182a.

M S Shim , Y N Xia . . Angew Chem Int Ed , 2013 . 52 ( 27 ): 6926 - 6929 . DOI:10.1002/anie.201209633http://doi.org/10.1002/anie.201209633.

X F Zhou , X R Liu , B X Zhao , X Liu , DC Zhu , NS Qiu , Q Zhou , Y Piao , Z X Zhou , J B Tang , YQ Shen . . J Control Release , 2016 . 234 90 - 97 . DOI:10.1016/j.jconrel.2016.05.042http://doi.org/10.1016/j.jconrel.2016.05.042.

W X Wang , C Alexander . . Angew Chem Int Ed , 2008 . 47 ( 41 ): 7804 - 7806 . DOI:10.1002/anie.v47:41http://doi.org/10.1002/anie.v47:41.

R J Amir , N Pessah , M Shamis , D Shabat . . Angew Chem Int Ed , 2003 . 115 ( 37 ): 4632 - 4637 . DOI:10.1002/(ISSN)1521-3757http://doi.org/10.1002/(ISSN)1521-3757.

M Avital-Shmilovici , D Shabat . . Bioorg Med Chem Lett , 2009 . 19 ( 14 ): 3959 - 3962 . DOI:10.1016/j.bmcl.2009.03.002http://doi.org/10.1016/j.bmcl.2009.03.002.

M Shamis , D Shabat . . Chem Eur J , 2007 . 13 ( 16 ): 4523 - 4528 . DOI:10.1002/(ISSN)1521-3765http://doi.org/10.1002/(ISSN)1521-3765.

R J Amir , M Popkov , R A Lerner , C F Barbas , D Shabat . . Angew Chem Int Ed , 2005 . 117 ( 28 ): 4452 - 4455 . DOI:10.1002/(ISSN)1521-3757http://doi.org/10.1002/(ISSN)1521-3757.

B Sauerbrei , V Jungmann , H Waldmann . . Angew Chem Int Ed , 1998 . 37 ( 8 ): 1143 - 1146 . DOI:10.1002/(ISSN)1521-3773http://doi.org/10.1002/(ISSN)1521-3773.

M Shamis , H N Lode , D Shabat . . J Am Chem Soc , 2004 . 126 ( 6 ): 1726 - 1731 . DOI:10.1021/ja039052phttp://doi.org/10.1021/ja039052p.

更多指标>

浏览量

下载量

CSCD

文章被引用时，请邮件提醒。

提交

工具集

关联资源

基于胺基与环氧的开环反应构建医用阳离子聚合物