基于透明质酸修饰的磁性碳纳米管靶向给药系统的研究

发布时间：2018-05-01 17:44

本文选题：碳纳米管 + 透明质酸　；参考：《郑州大学》2014年硕士论文

【摘要】：碳纳米管具有良好的细胞膜穿透性，并且可通过吸附、化学偶联或者包合作用携带化学药物或者生物大分子穿过细胞膜，进入细胞或组织，在传递药物方面有着潜在的优势。然而，未加修饰的碳纳米管表面疏水性强，在细胞中易聚集，毒性较大，，不适合作为药物载体。本课题旨在通过表面共价修饰的方法，将磁性Fe3O4纳米粒和具有肿瘤靶向性和亲水性的透明质酸多糖接枝到碳纳米管上，得到一种高水溶性、低毒性的磁性碳纳米管，通过物理吸附（π-π）堆积的作用装载化疗药物多西他赛（Docetaxel，DTX），构建一个水溶性良好的磁靶向给药系统，并以人乳腺癌MCF-7细胞和S180肉瘤荷瘤小鼠为对象考察其体内外抗肿瘤活性。首先，通过一系列的羧基化、氨基化以及还原反应，将Fe3O4纳米粒负载到碳纳米管上，得到超顺磁性的碳纳米管，再通过酯化反应将分子量14000~20000的透明质酸接枝到碳纳米管表面，得到水溶性、形态、粒径、电位均良好的磁性碳纳米管（CNTs-Fe3O4-HA）。并通过傅里叶变换红外光谱(FT-IR)、激光纳米粒度分析仪(DLS)、振动样品磁强计（VSM）以及透射电子显微镜（TEM）等对其合成过程及磁性行为进行表征，结果表明载体系统制备成功且具有良好的超顺磁性。其次，以合成的CNTs-Fe3O4-HA为载体，DTX为模型药物，通过物理吸附π-π堆积的作用构建CNTs-Fe3O4-HA/DTX给药系统，通过单因素考察优化处方设计，所得的CNTs-Fe3O4-HA/DTX给药系统载药率高达160%，所得制剂药物浓度高达2.3mg·ml-1，其在各个介质中可长期稳定存在，具有作为纳米给药系统的潜力。本研究以人乳腺癌MCF-7细胞为实验对象考察该给药系统的体外抗肿瘤活性。细胞毒性实验表明CNTs-Fe3O4-HA对细胞无明显毒性，而与DTX相比，CNTs-Fe3O4-HA/DTX在作用时间为72h时表现出较强的细胞抑制，表明该给药系统具有一定的缓释作用。细胞摄取实验表明CNTs-Fe3O4-HA可高效率的穿透细胞膜，促进了DTX的跨膜转运。此外，以S180肉瘤荷瘤小鼠为模型动物，考察CNTs-Fe3O4-HA/DTX给药系统的组织分布以及体内抗肿瘤活性。组织分布实验表明，CNTs-Fe3O4-HA可提高药物在肿瘤部位的浓集，降低DTX的肺毒性，且在外加磁场作用下长期连续给药后可有效提药物在肿瘤部位的浓度，降低DTX在肺中的分布，且在外加磁场作用下长期连续给药后可显著提高药物在肿瘤部位的分布，该结果表明，CNTs-Fe3O4-HA/DTX在体内具有显著的磁靶向能力。体内抗肿瘤实验表明，CNTs-Fe3O4-HA/DTX给药系统提高了DTX的抗肿瘤活性，且在外加磁场作用下，CNTs-Fe3O4-HA/DTX给药系统对肿瘤体积的增长抑制更加明显。
[Abstract]:Carbon nanotubes (CNTs) have good cell membrane penetration and have potential advantages in drug delivery by means of adsorption, chemical coupling or inclusion cooperation through cell membranes or biological macromolecules carrying chemical drugs or biological macromolecules. However, unmodified carbon nanotubes have strong hydrophobicity, easy aggregation and toxicity in cells, so they are not suitable as drug carriers. The aim of this study was to graft magnetic Fe3O4 nanoparticles and hyaluronic acid polysaccharides with tumor targeting and hydrophilicity onto carbon nanotubes by surface covalent modification to obtain a highly water-soluble and low-toxic magnetic carbon nanotubes. The chemotherapeutic drug docetaxelon DTX was loaded by physical adsorption (蟺-蟺) to construct a water-soluble magnetic targeting drug delivery system. The antitumor activity of human breast cancer MCF-7 cells and S180 sarcoma bearing mice was investigated in vitro and in vivo. First, the superparamagnetic carbon nanotubes were prepared by loading Fe3O4 nanoparticles onto carbon nanotubes through a series of carboxylation, amination and reduction reactions, and then grafted hyaluronic acid with molecular weight 14000,000000 onto the surface of carbon nanotubes by esterification. A magnetic carbon nanotube CNTs-Fe _ 3O _ 4-HAN _ 4 with good water solubility, morphology, particle size and potential was obtained. The synthesis process and magnetic behavior were characterized by Fourier transform infrared spectroscopy (FTIR), laser nanocrystalline analyzer (DLSX), vibrating sample magnetometer (VSM) and transmission electron microscope (TEM). The results showed that the carrier system was successfully prepared and had good superparamagnetic properties. Secondly, the drug delivery system of CNTs-Fe3O4-HA was constructed by physical adsorption of 蟺-蟺 accumulation, and the prescription design was optimized by single factor investigation. The drug loading rate of the CNTs-Fe3O4-HA/DTX drug delivery system is as high as 160 and the drug concentration of the preparation is as high as 2.3mg ml-1, which can exist in various media for a long time and has the potential as a nanometer drug delivery system. In this study, human breast cancer MCF-7 cells were used to investigate the antitumor activity of the drug delivery system in vitro. The cytotoxicity test showed that CNTs-Fe3O4-HA had no obvious cytotoxicity, but CNTs-Fe3O4-HA / DTX exhibited strong cell inhibition at 72 h compared with DTX, indicating that the drug delivery system had a certain sustained release effect. Cell uptake assay showed that CNTs-Fe3O4-HA could penetrate the cell membrane efficiently and promote the transmembrane transport of DTX. In addition, S180 sarcoma bearing mice were used as model animals to investigate the tissue distribution and antitumor activity of CNTs-Fe3O4-HA/DTX delivery system. The results of tissue distribution showed that CNTs-Fe _ 3O _ 4-HA could increase the concentration of DTX in the tumor site and decrease the pulmonary toxicity of DTX. The concentration of CNTs-Fe _ 3O _ 4-HA in the tumor site and the distribution of DTX in lung could be effectively extracted after long-term continuous administration under the action of external magnetic field. The results showed that CNTs-Fe _ 3O _ 4-HA / DTX had significant magnetic targeting ability in vivo. In vivo antitumor experiments showed that CNTs-Fe3O4-HA / DTX administration system increased the antitumor activity of DTX, and CNTs-Fe3O4-HA / DTX system inhibited tumor volume more obviously under the action of external magnetic field.
【学位授予单位】：郑州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R943

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