酶响应的树枝状大分子键合物在体外肿瘤模型中的渗透行为研究

陈思亲; 周泉; 相佳佳; 周珠贤; 唐建斌; 申有青

doi:10.11777/j.issn1000-3304.2020.20257

您当前的位置：

首页 >

文章列表页 >

酶响应的树枝状大分子键合物在体外肿瘤模型中的渗透行为研究

研究论文 | 更新时间：2021-05-07

- 酶响应的树枝状大分子键合物在体外肿瘤模型中的渗透行为研究
- Evaluation of in Vitro Tumor Penetration Behavior of Enzyme-responsive Dendrimer-drug Conjugate
- 高分子学报 2021年52卷第5期页码：477-488
- 作者机构：
  
  1.浙江大学化学工程与生物工程学院生物纳米工程中心生物质化工教育部重点实验室　杭州 310027
  2.浙江大学杭州国际科创中心　杭州 311200
- 作者简介：
  
  E-mail: shenyq@zju.edu.cn You-qing Shen, E-mail: shenyq@edu.cn
- 基金信息：
  
  国家自然科学基金(基金号 51833008，51522304，21704090)、浙江省重点研发计划(项目号 2020C01123)和中国博士后科学基金(基金号 2019M662059)资助项目()
- DOI：10.11777/j.issn1000-3304.2020.20257
  中图分类号：
- 纸质出版日期：2021-5-3，
  
  网络出版日期：2021-2-8，
  
  收稿日期：2020-11-28，
  
  修回日期：2020-12-24，
扫描看全文
陈思亲, 周泉, 相佳佳, 周珠贤, 唐建斌, 申有青. 酶响应的树枝状大分子键合物在体外肿瘤模型中的渗透行为研究[J]. 高分子学报, 2021,52(5):477-488.

Si-qin Chen, Quan Zhou, Jia-jia Xiang, Zhu-xian Zhou, Jian-bin Tang, You-qing Shen. Evaluation of in Vitro Tumor Penetration Behavior of Enzyme-responsive Dendrimer-drug Conjugate[J]. Acta Polymerica Sinica, 2021,52(5):477-488.
陈思亲, 周泉, 相佳佳, 周珠贤, 唐建斌, 申有青. 酶响应的树枝状大分子键合物在体外肿瘤模型中的渗透行为研究[J]. 高分子学报, 2021,52(5):477-488. DOI： 10.11777/j.issn1000-3304.2020.20257.

Si-qin Chen, Quan Zhou, Jia-jia Xiang, Zhu-xian Zhou, Jian-bin Tang, You-qing Shen. Evaluation of in Vitro Tumor Penetration Behavior of Enzyme-responsive Dendrimer-drug Conjugate[J]. Acta Polymerica Sinica, 2021,52(5):477-488. DOI： 10.11777/j.issn1000-3304.2020.20257.

摘要

实体肿瘤组织中固有的高渗透压、高细胞密度和乏血供等生物屏障导致纳米药物难以在肿瘤组织中浸润，从而难以渗透到肿瘤内部发挥治疗作用. 为了克服上述纳米药物的被动扩散瓶颈，提升其在肿瘤组织中的渗透效果，本文设计了一种基于主动转胞吞作用来实现跨细胞传递和肿瘤渗透的纳米载药系统. 利用

-谷氨酰胺转移酶(GGT)响应的两性离子基团(BGA)修饰了以喜树碱(CPT)为核心的第四代树枝状大分子(G4/CPT)，制备了一种具有精准结构和肿瘤特异性酶响应电荷反转的药物-树枝状大分子键合物(G4/CPT-BGA)，其分子量为20 kDa，粒径约为5 nm，表面电势约为−2 mV. 研究发现G4/CPT-BGA能够被GGT催化产生由负到正的电荷反转，并且能够水解释放出所携载的化疗药物喜树碱，从而有效杀伤肿瘤细胞. 通过流式细胞术实验和激光共聚焦显微镜证明了G4/CPT-BGA能通过小窝蛋白介导的细胞内吞被肿瘤细胞摄取，随后通过高尔基体介导的细胞外排途径被释放出细胞，由此通过这种迭代不断的“内吞-外排”作用(转胞吞)实现跨细胞传递. 最后，通过激光共聚焦显微镜观察G4/CPT-BGA在三维肿瘤球中的浸润效果，证明了G4/CPT-BGA能够基于主动转胞吞作用实现在肿瘤球中的高效渗透作用. 因此，用GGT响应的两性离子基团对树枝状大分子的端基进行修饰，能赋予该载体主动转胞吞作用的功能，从而克服被动转运过程中的扩散障碍，为设计其他具有肿瘤增强渗透能力的抗癌纳米药物递送系统提供了可能.

Abstract

The limited distribution of anticancer nanodrugs remains a bottle-neck for their therapeutic effect. Several strategies

such as surface modification

photothermal activation

and microenvironment modulation

have been studied to improve the penetration of anticancer drugs in solid tumours. However

the inherent high osmotic pressure

high cell density

lack of blood supply

and other biological barriers in solid tumors make it difficult for nanomedicine to infiltrate in the tumors

thus inaccessible to the distal cells to exert an effective therapeutic effect. Therefore

the way to deliver sufficient drugs to infiltrate in a tumour is a key problem that should be solved quickly. Herein

we constructed a molecularly precise polylysine-dendrimer-drug conjugate with

-glutamyl transpeptidase (GGT)-sensitive termini and obtained a zwitterionic

-glutamyl functionalized dendrimer-drug conjugate

i.e.

G4/CPT-BGA. The results showed the molecular weight of G4/CPT-BGA dendrimer was 20 kDa with a small size of 5 nm and a moderate surface charge of −2 mV. The G4/CPT-BGA could undergo rapid GGT-triggered charge-reversal from zwitterionic to cationic

thereby quickly endocytosed by tumor cells

releasing the conjugated drug (CPT) to exert effective cytotoxicity. The

in vitro

endocytosis and exocytosis experiments showed that G4/CPT-BGA was able to traffic in cells through a caveolae-mediated pathway

then traffic out of cells

via

Golgi-apparatus

thus achieving active transcytosis for transcellular delivery. The tumor penetration ability of G4/CPT-BGA was investigated using three-dimensional multicellular spheroids

which showed G4/CPT-BGA could evenly distribute throughout the spheroids

via

transcytosis-mediated active tumour infiltration. Therefore

a simple modification of the cationic dendrimer with GGT responsive zwitterionic

-glutamine enables the dendrimer to actively transcytosis across cells

thereby avoiding the paracellular diffusion obstacles

which may also be applicable for designing anticancer nanomedicine systems with enhanced penetration ability.

关键词

纳米药物树枝状大分子肿瘤渗透主动转胞吞作用γ-谷氨酰胺转移酶

Keywords

NanomedicineDendrimerTumor penetrationActive transcytosisγ-Glutamyl transpeptidase

references

Porche D J . J Assoc Nurses AIDS Care , 1996 . 7 ( 2 ): 55 - 59 . DOI:10.1016/S1055-3290(96)80016-1http://doi.org/10.1016/S1055-3290(96)80016-1 .

Kundranda M N, Niu J . Drug Des Devel Ther , 2015 . 9 3767 - 3777.

Zhang H J . Onco Targets Ther , 2016 . 9 3001 - 3007.

Cabral H, Kataoka K . J Control Release , 2014 . 190 465 - 476 . DOI:10.1016/j.jconrel.2014.06.042http://doi.org/10.1016/j.jconrel.2014.06.042 .

Shi J J, Kantoff P W, Wooster R, Farokhzad O C . Nat Rew Cancer , 2017 . 17 ( 1 ): 20 - 37 . DOI:10.1038/nrc.2016.108http://doi.org/10.1038/nrc.2016.108 .

Adiseshaiah P P, Crist R M, Hook S S, McNeil S E . Nat Rev Clin Oncol , 2016 . 13 ( 12 ): 750 - 765 . DOI:10.1038/nrclinonc.2016.119http://doi.org/10.1038/nrclinonc.2016.119 .

Chan W C W . Acc Chem Res , 2017 . 50 ( 3 ): 627 - 632 . DOI:10.1021/acs.accounts.6b00629http://doi.org/10.1021/acs.accounts.6b00629 .

Sun Q H, Sun X R, Ma X P, Zhou Z X, Jin E, Zhang B, Shen Y, van Kirk E A, Murdoch W J, Lott J R, Lodge T P, Radosz M, Zhao Y L . Adv Mater , 2014 . 26 ( 45 ): 7615 - 7621 . DOI:10.1002/adma.201401554http://doi.org/10.1002/adma.201401554 .

Sun Q H, Zhou Z X, Qiu N S, Shen Y Q . Adv Mater , 2017 . 29 ( 14 ): 201606628 .

Gao Jing(高晶), Wang Weiqi(王伟奇), Yu Haijun(于海军) . Acta Polymerica Sinica(高分子学报) , 2019 . 50 ( 11 ): 1156 - 1166 . DOI:10.11777/j.issn1000-3304.2019.19133http://doi.org/10.11777/j.issn1000-3304.2019.19133 .

Sun Rui(孙瑞), Qiu Nasha(邱娜莎), Shen Youqing(申有青) . Acta Polymerica Sinica(高分子学报) , 2019 . 50 ( 6 ): 588 - 601.

Prabhakar U, Maeda H, Jain R K, Sevick-Muraca E M, Zamboni W, Farokhzad O C, Barry S T . Cancer Res , 2013 . 73 ( 8 ): 2412 - 2417 . DOI:10.1158/0008-5472.CAN-12-4561http://doi.org/10.1158/0008-5472.CAN-12-4561 .

Gao N, Bozeman E N, Qian W P, Wang L Y, Chen H Y, Lipowska M, Staley C A . Theranostics , 2017 . 7 ( 6 ): 1689 - 1704 . DOI:10.7150/thno.18125http://doi.org/10.7150/thno.18125 .

Primeau A J, Rendon A, Hedley D, Lilge L, Tannock I F . Clin Cancer Res , 2005 . 11 ( 24 ): 8782 - 8788.

Minchinton A I, Tannock I F . Nat Rev Cancer , 2006 . 6 ( 8 ): 583 - 592 . DOI:10.1038/nrc1893http://doi.org/10.1038/nrc1893 .

Zhou Q, Dong C Y, Fan W F, Jiang H P, Xiang J J, Qiu N S, Piao Y, Xie T, Luo YW, Li Z C, Liu F S, Shen Y Q . Biomaterials , 2020 . 240 119902 DOI:10.1016/j.biomaterials.2020.119902http://doi.org/10.1016/j.biomaterials.2020.119902 .

Sugahara K N, Teesalu T, Karmali P P, Kotamraju V R, Agemy L, Greenwald D R, Ruoslahti E . Science , 2010 . 328 ( 5981 ): 1031 - 1035 . DOI:10.1126/science.1183057http://doi.org/10.1126/science.1183057 .

Ruoslahti E . Adv Drug Deliv Rev , 2017 . 110 3 - 12.

Sun W J, Hu Q Y, Ji W Y, Wright G, Gu Z . Physiol Rev , 2017 . 97 ( 1 ): 189 - 225 . DOI:10.1152/physrev.00015.2016http://doi.org/10.1152/physrev.00015.2016 .

Li H J, Du J Z, Du X J, Xu C F, Sun C Y, Wang H X, Cao Z T . Proc Natl Acad Sci USA , 2016 . 113 ( 15 ): 4164 - 4169 . DOI:10.1073/pnas.1522080113http://doi.org/10.1073/pnas.1522080113 .

Dewhirst M W, Secomb T W . Nat Rew Cancer , 2017 . 17 ( 12 ): 738 - 750 . DOI:10.1038/nrc.2017.93http://doi.org/10.1038/nrc.2017.93 .

Yin Q, Tang L, Cai K M, Yang X J, Yin L C, Zhang Y F, Dobrucki L W . Biomater Sci , 2018 . 6 ( 5 ): 1189 - 1200 . DOI:10.1039/C8BM00149Ahttp://doi.org/10.1039/C8BM00149A .

Wiley D T, Webster P, Gale A, Davis M E . Proc Natl Acad Sci USA , 2013 . 110 ( 21 ): 8662 - 8667 . DOI:10.1073/pnas.1307152110http://doi.org/10.1073/pnas.1307152110 .

Martin-Latil S, Gnadig N F, Mallet A, Desdouits M, Guivel-Benhassine F, Jeannin P, Prevost M C, Schwartz O, Gessain A, Ozden S, Ceccaldi P E . Blood , 2012 . 120 ( 3 ): 572 - 580 . DOI:10.1182/blood-2011-08-374637http://doi.org/10.1182/blood-2011-08-374637 .

Wang G W, Zhou Z X, Zhao Z H, Li Q Y, Wu Y L, Yan S, Shen Y Q, Huang P T . ACS Nano , 2020 . 14 ( 4 ): 4890 - 4904 . DOI:10.1021/acsnano.0c00974http://doi.org/10.1021/acsnano.0c00974 .

Zhou Q, Shao S Q, Wang J Q, Xu C H, Xiang J J, Piao Y, Zhou Z X, Yu Q S, Tang J B, Liu X R, Gan Z H, Mo R, Gu Z, Shen Y Q . Nat Nanotechnol , 2019 . 14 ( 8 ): 799 - 809 . DOI:10.1038/s41565-019-0485-zhttp://doi.org/10.1038/s41565-019-0485-z .

Wang G W, Wu B H, Li Q Y, Chen S Q, Jin X Q, Liu Y J, Zhou Z X, Shen Y Q, Huang P T . Small , 2020 . 16 ( 44 ): e2004172 DOI:10.1002/smll.202004172http://doi.org/10.1002/smll.202004172 .

Suzuki H, Bae Y H . Biomaterials , 2016 . 98 120 - 130 . DOI:10.1016/j.biomaterials.2016.04.037http://doi.org/10.1016/j.biomaterials.2016.04.037 .

Tang Zhaohui(汤朝晖), Chen Xuesi(陈学思) . Acta Polymerica Sinica(高分子学报) , 2019 . 50 ( 6 ): 543 - 552.

Jin Q, Deng Y Y, Chen X H, Ji J . ACS Nano , 2019 . 13 ( 2 ): 954 - 977.

Shao S Q, Zhou Q, Si J X, Tang J B, Liu X R, Wang M, Gao J Q, Wang K, Xu R Z, Shen Y Q . Nat Biomed Eng , 2017 . 1 ( 9 ): 745 - 757 . DOI:10.1038/s41551-017-0130-9http://doi.org/10.1038/s41551-017-0130-9 .

Liu Chang(刘畅), Chen Yuxin(陈宇欣), Wang Jiangfan(王江帆), Luo Xuan(罗萱), Huang Yudi(黄宇迪), Xu Jinlei(徐瑾蕾), Yan Guoping(鄢国平), Chen Si(陈思), Zhang Xianzheng(张先正) . Acta Polymerica Sinica(高分子学报) , 2018 . ( 6 ): 682 - 691 . DOI:10.11777/j.issn1000-3304.2017.17335http://doi.org/10.11777/j.issn1000-3304.2017.17335 .

Zhou Z X, Ma X P, Murphy C J, Jin E L, Sun Q H, Shen Y Q, van Kirk E A, Murdoch W J . Angew Chem Int Ed , 2014 . 53 ( 41 ): 10949 - 10955 . DOI:10.1002/anie.201406442http://doi.org/10.1002/anie.201406442 .

Forest V, Cottier M, Pourchez J . Nano Today , 2015 . 10 ( 6 ): 677 - 680 . DOI:10.1016/j.nantod.2015.07.002http://doi.org/10.1016/j.nantod.2015.07.002 .

Yazdani S, Jaldin-Fincati J R, Pereira R V S, Klip A . Traffic , 2019 . 20 ( 6 ): 390 - 403 . DOI:10.1111/tra.12645http://doi.org/10.1111/tra.12645 .

Frohlich E . Int J Nanomedicine , 2012 . 7 5577 - 5591.

Liu H M, Wang Y, Wang M M, Xiao J R, Cheng Y Y . Biomaterials , 2014 . 35 ( 20 ): 5407 - 5413 . DOI:10.1016/j.biomaterials.2014.03.040http://doi.org/10.1016/j.biomaterials.2014.03.040 .

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

纳米药物在非肌层浸润性膀胱癌灌注治疗中的应用

聚谷氨酸接枝聚乙二醇抗肿瘤药物靶向输送系统

高分子抗肿瘤纳米药物的挑战与发展

基于TACMAA化学的肿瘤微酸性环境响应高分子纳米药物载体

基于含POSS-dendrimer超强液晶凝胶的柔性可拉伸液晶光散射显示器