靶向纳米钻石-甲氨蝶呤药物体系制备及与MCF-7细胞作用

doi:10.7503/cjcu20190247

高等学校化学学报 ›› 2019, Vol. 40 ›› Issue (9): 1998.doi: 10.7503/cjcu20190247

靶向纳米钻石-甲氨蝶呤药物体系制备及与MCF-7细胞作用

李林^1,^*(),许鑫汝¹,李英奇³,张彩凤^1,²

^1. 太原师范学院化学系, 晋中 030619
^2. 山西省腐植酸工程技术研究中心, 晋中 030619
^3. 山西大学化学化工学院, 太原 030006

收稿日期:2019-04-28 出版日期:2019-09-10 发布日期:2019-08-20
通讯作者: 李林 E-mail:lilin@tynu.edu.cn
基金资助:
山西省青年科技研究基金(201801D221440);山西省应用基础研究基金(201801D121257);太原师范学院大学生科技创新项目(2018503 CXCY1850)

Preparation of Targeting Nanodiamond-metaminopterone Drug System and Its Interaction with MCF-7 Cells ^†

LI Lin^1,^*(),XU Xinru¹,LI Yingqi³,ZHANG Caifeng^1,²

^1. Department of Chemistry, Taiyuan Normal University, Jinzhong, 030619, China
^2. Humic Acid Engineering and Technology Research Center of Shanxi Province, Jinzhong 030619, China
^3. College of Chemistry and Chemical Engineering, Shanxi University, Taiyuan 030006, China

Received:2019-04-28 Online:2019-09-10 Published:2019-08-20
Contact: LI Lin E-mail:lilin@tynu.edu.cn
Supported by:
? Supported by the Shanxi Youth Science and Technology Research Fund Project, China(201801D221440);the Shanxi Provincial Applied Fundamental Research Fund Project, China(201801D121257);the Taiyuan Normal University College Students Science and Technology Innovation Project, China(No. 2018503 CXCY1850)(2018503 CXCY1850)

摘要/Abstract

摘要：

以羧基化纳米钻石(ND-COOH)为基体, 通过共价键合方法将聚乙二醇二胺(H₂N-PEG-NH₂)、叶酸(FA)和缩水甘油(GLY)偶联于ND-COOH表面, 赋予纳米钻石载体较好的水溶分散性和靶向性, 借助氢键和范德华力等作用力负载甲氨蝶呤(MTX), 得到靶向纳米钻石-聚乙二醇二胺-叶酸/缩水甘油/甲氨蝶呤(ND-PEG-FA/GLY/MTX NPF/G/M)纳米药物体系. 采用透射电子显微镜、 X射线能量色散谱、粒径及电位测试证实已制备NPF/G/M. 体外释药发现NPF/G/M在肿瘤环境(pH=5.5)中的药物释放量为正常生理环境(pH=7.4)中的3倍, 表明其具有良好的药物输送特性. 此外, 利用流式细胞术和MTT毒性测试探究了MCF-7细胞摄取NPF/G/M的机制及动力学特性和细胞毒性, 结果表明NPF/G/M以依赖能量、温度、网格蛋白、小窝蛋白和叶酸受体介导的机制进入细胞, 从而将药物缓慢释放于细胞内, 进而诱导细胞凋亡. 研究结果表明, NPF/G/M可作为一种良好的药物输送体系, 为其应用于乳腺癌的临床治疗提供理论参考.

关键词: 纳米钻石, 聚乙二醇二胺, 叶酸, 缩水甘油, 甲氨蝶呤, 靶向纳米药物

Abstract:

We fabricated a targeted nanodiamond drug system(NPF/G/M) through physical adsorption of methotrexate(MTX) using carboxylated nanodiamonds(ND-COOH) modified with polyethylene glycol diamine(H₂N-PEG-NH₂), folic acid(FA) and glycidol(GLY) as carriers. The preparation of NPF/G/M was verified by morphological observation, energy dispersive spectroscopy(EDS) of the elemental analysis, zeta potential and particle size. The results showed that NPF/G/M can be effectively released in the tumor site, which is three times than in the physiological environment(pH=7.4). Relying on the high affinity between folate and folate receptors, NPF/G/M was uptaken by tumor cells through energy, temperature, clathrin, caveolin-dependent and folate receptor-mediated endocytosis. Interestingly, toxicity tests showed that NPF/G/M can slowly release the drug into the cells and induce cell apoptosis. The above results indicate that NPF/G/M can be used as a good drug delivery system, which will provide a theoretical reference for clinical application in breast cancer.

Key words: Nanodiamond, Polyethylene glycol diamine, Folic acid, Glycidol, Methotrexate, Targeted nanodrug

中图分类号:

O631
O611.4

TrendMD:

李林, 许鑫汝, 李英奇, 张彩凤. 靶向纳米钻石-甲氨蝶呤药物体系制备及与MCF-7细胞作用. 高等学校化学学报, 2019, 40(9): 1998.

LI Lin, XU Xinru, LI Yingqi, ZHANG Caifeng. Preparation of Targeting Nanodiamond-metaminopterone Drug System and Its Interaction with MCF-7 Cells ^†. Chem. J. Chinese Universities, 2019, 40(9): 1998.

图/表 10

Fig.1 Structures of FA-NHS(A), GLY(B) and MTX(C)

Scheme 1 Synthesis of ND-PEG-FA/GLY/MTX nanomedicine

Fig.2 TEM images of ND(A), NPF/G(B) and NPF/G/M(C)

Fig.3 EDS spectra of ND(A), NPF/G(B) and NPF/G/M(C) Inset: SEM images of corresponding nanomaterials.

Table 1 Mass fraction of major elements of various nanomaterials

Nanoparticles	Mass fraction of major element(%)
Nanoparticles	C	N	O
ND	97.01	0	2.99
NPF/G	81.90	7.64	10.47
NPF/G/M	75.38	8.56	16.07

Fig.4 Particle size(A) and Zeta potential(B) of various nanomaterials

Fig.5 Release curves of ND-PEG-FA/GLY/MTX nanomedicine at pH=5.5(a) and 7.4(b)

Fig.6 Effect of uptake quality of ND-PEG-FA/GLY/MTX by MCF-7 cells (A) The effect of different concentrations of free FA on the amount of ND-PEG-FA/GLY/MTX uptaken by MCF-7 cellsby flow cytometry; (B) the mechanism of ND-PEG-FA/GLY/MTX nanomedicine uptaken by MCF-7 cells; (C) dynamics of the nanomedicine ND-PEG-FA/GLY/MTX uptaken by MCF-7 cells.

Fig.7 Microscopy images of MCF-7 cells incubated with different nanoparticles for 48 h(A1—A4) and 72 h(B1—B4) (A1, B1) Control; (A2, B2) NPF/G;(A3, B3) NPF/G/M; (A4, B4) MTX.

Fig.8 Effects of various nanomaterials on MCF-7 cells viability by MTT assay with different time

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靶向纳米钻石-甲氨蝶呤药物体系制备及与MCF-7细胞作用

Preparation of Targeting Nanodiamond-metaminopterone Drug System and Its Interaction with MCF-7 Cells ^†

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靶向纳米钻石-甲氨蝶呤药物体系制备及与MCF-7细胞作用

Preparation of Targeting Nanodiamond-metaminopterone Drug System and Its Interaction with MCF-7 Cells †

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Preparation of Targeting Nanodiamond-metaminopterone Drug System and Its Interaction with MCF-7 Cells ^†