EcN共价PSMA/PELA载药微球的制备及其靶向肿瘤低氧环境的初步研究

罗晓明; 毛皓月; 杨铃; 韩彬; 苏春丽; 宋海星

doi:10.16016/j.1000-5404.201811194

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EcN共价PSMA/PELA载药微球的制备及其靶向肿瘤低氧环境的初步研究

基础医学 | 更新时间：2026-01-09

- EcN共价PSMA/PELA载药微球的制备及其靶向肿瘤低氧环境的初步研究
- Preparation of EcN-conjugated drug-loaded PSMA/PELA micro-particles and their ability to target hypoxic tumor environment
- 第三军医大学学报 2019年41卷第10期页码：947-954
- 作者机构：
  
  1.610500 成都，成都医学院：公共卫生学院预防医学教研室
  2.610500 成都，成都医学院：生物技术学院实验教学中心
- 作者简介：
  
  [ 宋海星，E-mail: shx@cmc.edu.cn SONG Haixing, E-mail: shx@cmc.edu.cn
- 基金信息：
  
  国家自然科学基金青年科学基金(81501581);四川省科技厅重点研发项目(2017GZ0426)
- DOI：10.16016/j.1000-5404.201811194
  中图分类号： R378.2+1;R730.53;R944.9
- 收稿：2018-11-29，
  
  修回：2019-01-02，
  
  纸质出版：2019-05-30
- 稿件说明：
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罗晓明, 毛皓月, 杨铃, 等. EcN共价PSMA/PELA载药微球的制备及其靶向肿瘤低氧环境的初步研究[J]. 第三军医大学学报, 2019,41(10):947-954.

Xiaoming LUO, Haoyue MAO, Ling YANG, et al. Preparation of EcN-conjugated drug-loaded PSMA/PELA micro-particles and their ability to target hypoxic tumor environment[J]. Journal of Third Military Medical University, 2019, 41(10): 947-954.
罗晓明, 毛皓月, 杨铃, 等. EcN共价PSMA/PELA载药微球的制备及其靶向肿瘤低氧环境的初步研究[J]. 第三军医大学学报, 2019,41(10):947-954. DOI： 10.16016/j.1000-5404.201811194.

Xiaoming LUO, Haoyue MAO, Ling YANG, et al. Preparation of EcN-conjugated drug-loaded PSMA/PELA micro-particles and their ability to target hypoxic tumor environment[J]. Journal of Third Military Medical University, 2019, 41(10): 947-954. DOI： 10.16016/j.1000-5404.201811194.

摘要

目的

将Escherichia coli Nissle 1917(EcN)与聚苯乙烯马来酸酐(poly styrene-co-maleic anhydride

PSMA)/聚DL-乳酸-聚乙二醇共聚物[(poly (DL-Lactic acid) and poly ethyleneglycol block copolymer

PELA]聚合物载药微粒共价结合，研究其靶向肿瘤低氧环境的效果。

方法

溶剂挥发法制备不同混合比例(质量比为7 :3和5 :5)的PSMA和PELA携载阿霉素(Dox)的PSMA/PELA微粒；通过EDC和NHS催化反应使得EcN表面的氨基和PSMA/PELA载药微粒表面的酸酐发生共价结合，平板计数法表征接枝后EcN的生长活性；利用HepG2细胞体外构建3D肿瘤模型，研究EcN共价PSMA/PELA载药微粒在肿瘤低氧区域的渗透效果；小鼠肝癌细胞H22建立皮下肿瘤模型，体内验证其靶向肿瘤低氧区域的效果。

结果

成功制备载Dox的PSMA/PELA微粒(DMP

7/3

和DMP

5/5

)，两种微粒球形规则，粒径700~800 nm；DMP

7/3

和DMP

5/5

载药量分别为(4.27±0.19)%和(3.94±0.11)%，包裹效率分别为(85.4±2.6)%和(78.8±3.5)%，载药量和包裹效率差异无统计学意义；CLSM结果显示，EcN成功接枝在DMP

7/3

表面，且接枝后的细菌具有生长活力；体外渗透实验研究表明，与EcN未共价接枝的DMP比较，EcN修饰的聚合物载药微粒(EcN@DMP

7/3

)可较多地渗透至肿瘤内部；体内分布实验证实，静脉给药4 h和24 h后，EcN@DMP

7/3

治疗组肿瘤组织内Dox浓度分别是DMP

7/3

治疗组浓度的3倍和6倍(

0.01)。

结论

通过EDC和NHS催化反应方法可使EcN表面的氨基和PSMA/PELA载药微粒表面的酸酐发生共价反应，且反应后的EcN依然维持其生长活性，能有效驱动PSMA/PELA载药微粒靶向肿瘤内部低氧区域。

Abstract

Objective

To conjugate

Escherichia coli

Nissle 1917 (EcN) with drug-loaded poly styrene-co-maleic anhydride(PSMA)/poly (DL-lactic acid) and poly ethyleneglycol block copolymer (PELA) polymer micro-particles and investigate the ability of the micro-particles to target the hypoxic tumor environment.

Methods

PSMA and PELA were mixed at the ratio of 7 :3 or 5 :5 (m/m) to prepare doxorubicin (Dox)-loaded PSMA/PELA micro-particles through solvent evaporation method. The covalent binding was achieved by EDC-NHS catalytic reaction of the carboxyl groups on the surface of EcN with the amino groups on the surface of PSMA/PELA drug-loaded micro-particles. In an

in vitro

3D tumor model constructed using HepG2 cells

we investigated the permeation of EcN-conjugated PSMA/PELA drug-loaded micro-particles into the hypoxic tumor area. We further tested the performance of the drug-loaded micro-particles for targeting the hypoxic tumor environment in a mouse model bearing subcutaneous hepatoma H22 cell xenograft.

Results

We successfully prepared Dox-loaded micro-particles with different PSMA/PELA ratios (DMP

7/3

and DMP

5/5

) using solvent e

vaporation method

and both of the micro-particles showed a regular spherical shape with an average size of 700 to 800 nm. DMP

7/3

and DMP

5/5

micro-particles had comparable Dox-loading efficiency [(4.27±0.19)%

(3.94±0.11)%

]

and encapsulation efficiency [(85.4± 2.6)%

(78.8±3.5)%

]

. Confocal laser scanning microscopy showed that EcN was successfully conjugated on the surface DMP

7/3

micro-particles. Compared with DMP microparticles without EcN conjugation

the EcN-conjugated micro-particles (EcN@DMP

7/3

) permeated efficiently into the central area in the

in vitro

3D tumor model. In the tumor-bearing mice

bio-distribution analyses revealed that at 4 and 24 h after intravenous administration

EcN@DMP

7/3

resulted in 3-folder and 6-folder increases in Dox accumulation in the tumors

respectively

as compared with the non-conjugated DMP

7/3

micro-particles (

0.01).

Conclusion

EDC-NHS catalytic reaction allows the amino groups on the outer surface of EcN to covalently bind to anhydride on the surface of drug-loaded PSMA/PELA particles without affecting the growth activity of EcN

which effectively drives the drug-loaded particles to target the hypoxic region inside the tumor.

关键词

Keywords

references

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