Preparation of EcN-conjugated drug-loaded PSMA/PELA micro-particles and their ability to target hypoxic tumor environment

Xiaoming LUO; Haoyue MAO; Ling YANG; Bin HAN; Chunli SU; Haixing SONG

doi:10.16016/j.1000-5404.201811194

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Preparation of EcN-conjugated drug-loaded PSMA/PELA micro-particles and their ability to target hypoxic tumor environment

Basic Medicine | 更新时间：2026-01-09

- Preparation of EcN-conjugated drug-loaded PSMA/PELA micro-particles and their ability to target hypoxic tumor environment
- Journal of Third Military Medical University Vol. 41, Issue 10, Pages: 947-954(2019)
- 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：
- 基金信息：
  
  National Natural Science Foundation for Young Scholars of China(81501581);Key Research and Development Project of Science and Technology Department of Sichuan Province(2017GZ0426)
- DOI：10.16016/j.1000-5404.201811194
  CLC： R378.2+1;R730.53;R944.9
- Received：29 November 2018，
  
  Revised：2019-01-02，
  
  Published：30 May 2019
- 稿件说明：
移动端阅览
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：

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

PEREIRA M C, RESHETNYAK Y K, ANDREEV O A. Advanced targeted nanomedicine[J]. J Biotechnol, 2015, 202: 88-97. DOI: 10.1016/j.jbiotec.2015.01.009.

PRABHU R H, PATRAVALE V B, JOSHI M D. Polymeric nanoparticles for targeted treatment in oncology: current insights[J]. Int J Nanomedicine, 2015, 10: 1001-1018. DOI: 10.2147/IJN.S56932.

SHA H, ZOU Z, XIN K, et al. Tumor-penetrating peptide fused EGFR single-domain antibody enhances cancer drug penetration into 3D multicellular spheroids and facilitates effective gastric cancer therapy[J]. J Control Release, 2015, 200: 188-200. DOI: 10.1016/j.jconrel.2014.12.039.

DAS M, DUAN W, SAHOO S K. Multifunctional nanoparticle-EpCAM aptamer bioconjugates:A paradigm for targeted drug delivery and imaging in cancer therapy[J]. Nanomedicine, 2015, 11(2): 379-389. DOI:10.1016/j.nano.2014.09.002.

DHANI N, FYLES A, HEDLEY D, et al. The clinical significance of hypoxia in human cancers[J]. Semin Nucl Med, 2015, 45(2): 110-121. DOI: 10.1053/j.semnuclmed.2014.11.002.

HONG M, ZHU S, JIANG Y, et al. Efficient tumor targeting of hydroxycamptothecin loaded PEGylated niosomes modified with transferrin[J]. J Control Release, 2009, 133(2): 96-102. DOI: 10.1016/j.jconrel.2008.09.005.

FELFOUL O, MOHAMMADI M, TAHERKHANI S, et al. Magneto-aerotactic bacteria deliver drug-containing nanoliposomes to tumour hypoxic regions[J]. Nat Nanotechnol, 2016, 11(11): 941-947. DOI: 10.1038/nnano.2016.137.

ZHOU S B, GRAVEKAMP C, BERMUDES D, et al. Tumour-targeting bacteria engineered to fight cancer[J]. Nat Rev Cancer, 2018, 18(12): 727-743. DOI: 10.1038/s41568-018-0070-z.

THAMM D H, KURZMAN I D, KING I, et al. Systemic administration of an attenuated, tumor-targeting Salmonella typhimurium to dogs with spontaneous neoplasia: phase I evaluation[J]. Clin Cancer Res, 2005, 11(13): 4827-4834. DOI: 10.1158/1078-0432.CCR-04-2510.

TOSO J F, GILL V J, HWU P, et al. Phase I study of the intravenous administration of attenuated Salmonella typhimurium to patients with metastatic melanoma[J]. J Clin Oncol, 2002, 20(1): 142-152. DOI: 10.1200/JCO.2002.20.1.142.

SOUZA É L, ELIAN S D, PAULA L M, et al. Escherichia coli strain Nissle 1917 ameliorates experimental colitis by modulating intestinal permeability, the inflammatory response and clinical signs in a faecal transplantation model[J]. J Med Microbiol, 2016, 65(3): 201-210. DOI: 10.1099/jmm.0.000222.

STRITZKER J, WEIBEL S, HILL P J, et al. Tumor-specific colonization, tissue distribution, and gene induction by probiotic Escherichia coli Nissle 1917 in live mice[J]. Int J Med Microbiol, 2007, 297(3): 151-162. DOI: 10.1016/j.ijmm.2007.01.008.

ZHOU S N, ZHANG M X, WANG J S. Tumor-targeted delivery of TAT-Apoptin fusion gene using Escherichia coli Nissle 1917 to colorectal cancer[J]. Med Hypotheses, 2011, 76(4): 533-534. DOI: 10.1016/j.mehy.2010.12.010.

SHI C L, GUO X, QU Q Q, et al. Actively targeted delivery of anticancer drug to tumor cells by redox-responsive star-shaped micelles[J]. Biomaterials, 2014, 35(30): 8711-8722. DOI: 10.1016/j.biomaterials.2014.06.036.

SIMS L B, HUSS M K, FRIEBOES H B, et al. Distribution of PLGA-modified nanoparticles in 3D cell culture models of hypo-vascularized tumor tissue[J]. J Nanobiotechnology, 2017, 15(1): 67. DOI: 10.1186/s12951-017-0298-x.

KRAMER M G, MASNER M, FERREIRA F A, et al. Bacterial therapy of cancer: promises, limitations, and insights for future directions[J]. Front Microbiol, 2018, 9: 16. DOI: 10.3389/fmicb.2018.00016.

LIU K, CHEN W J, YANG T T, et al. Paclitaxel and quercetin nanoparticles co-loaded in microspheres to prolong retention time for pulmonary drug delivery[J]. Int J Nanomed, 2017, 12: 8239-8255. DOI:10.2147/IJN.S147028.

KAMALY N, YAMEEN B, WU J, et al. Degradable controlled-release polymers and polymeric nanoparticles: mechanisms of controlling drug release[J]. Chem Rev, 2016, 116(4): 2602-2663. DOI: 10.1021/acs.chemrev.5b00346.

LOUISE MEYER R, ZHOU X F, TANG L, et al. Immobilisation of living bacteria for AFM imaging under physiological conditions[J]. Ultramicroscopy, 2010, 110(11): 1349-1357. DOI: 10.1016/j.ultramic.2010.06.010.

WANG C, TANG Z, ZHAO Y, et al. Three-dimensional in vitro cancer models: A short review[J]. Biofabrication, 2014, 6(2): 022001. DOI: 10.1088/1758-5082/6/2/022001.

WANG Y, MIRZA S, WU S, et al. 3D hydrogel breast cancer models for studying the effects of hypoxia on epithelial to mesenchymal transition[J]. Oncotarget, 2018, 9(63): 32191-32203. DOI: 10.18632/oncotarget.25891.

LUO X M, JIA G Q, SONG H X, et al. Promoting antitumor activities of hydroxycamptothecin by encapsulation into acid-labile nanoparticles using electrospraying[J]. Pharm Res, 2014, 31(1): 46-59. DOI: 10.1007/s11095-013-1130-4.

LUO C H, HUANG C T, SU C H, et al. Bacteria-mediated hypoxia-specific delivery of nanoparticles for tumors imaging and therapy[J]. Nano Lett, 2016, 16(6): 3493-3499. DOI: 10.1021/acs.nanolett.6b00262.

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