富勒烯—四氧化三铁纳米复合物磁靶向药物转运系统的制备及应用

发布时间：2018-05-13 08:37

本文选题：富勒烯 + 四氧化三铁　；参考：《郑州大学》2014年硕士论文

【摘要】：富勒烯（C60）具有独特的结构和特殊的光学性质，在药物转运和肿瘤光动力学治疗方面具有潜在应用价值，但其难溶性极大地限制了在生物医学领域中的应用。本课题研究目的是对富勒烯进行化学修饰，在其表面连接磁性四氧化三铁纳米粒（IONPs）和亲水性基团聚乙二醇2000（PEG2000），开发出新型磁性水溶性药物转运载体C60-IONP-PEG，并携带小分子光敏剂血卟啉单甲醚（HMME），构建一个具有良好的磁靶向能力、光疗能力和核磁成像能力的多功能肿瘤靶向给药系统C60-IONP-PEG/HMME。本课题首先构建了基于富勒烯衍生物的药物载体。利用Bingle环加成反应、酯水解反应、水热化学键合反应合成四氧化三铁负载的富勒烯（C60-IONP-COOH），利用聚乙二醇2000（PEG(2000)-NH2）的氨基与该富勒烯衍生物的羧基加成，制备出磁性的C60载体系统C60-IONP-PEG。使用傅里叶变换红外光谱验证该载体系统制备是否成功，使用透射电子显微镜、激光纳米粒度分析仪等进行形态表征，，使用振动样品磁测量仪来检测磁性，使用热重分析仪测定该系统中PEG的含量，使用3T核磁共振扫描仪考察该系统核磁成像能力。结果表明该磁性载体系统制备成功，C60-IONP-PEG具有高度的水溶性、明显的超顺磁性和显著的核磁成像能力，并且具有合适的粒径和电位，可作为优良的磁靶向药物转运载体。在磁靶向纳米制剂的制备中，以小分子光敏剂HMME为模型药物，通过单因素实验考察HMME与载体材料的投料比、超声时间、探超功率等因素对制剂载药量的影响，确定了拟构建肿瘤靶向给药系统C60-IONP-PEG/HMME的最佳处方工艺。使用透射电子显微镜、激光纳米粒度分析仪和紫外全波长扫描仪对该系统进行表征，结果表明C60-IONP-PEG能够高效负载HMME（载药率~125.3%），构建的给药系统具有良好的稳定性和大小均一的粒径，能够缓释药物，可持续释放药物达120小时。在体外抗肿瘤活性评价中，选用鼠源黑色素瘤（B16-F10）细胞作为研究对象，对HMME和C60协同增强光动力学治疗肿瘤的作用进行了研究，结果表明空白载体C60-IONP-PEG对B16-F10细胞无明显毒性，但在532nm激光照射下能够高效产生活性氧，可作为光敏剂应用于光动力学治疗中。此外，532nm激光照射后，C60-IONP-PEG/HMME体系中的C60和HMME均高效产生活性氧，协同增强光疗作用，共同杀伤肿瘤细胞，且具有浓度和光照强度依赖性。实验结果还表明C60-IONP-PEG/HMME具有良好的磁靶向性，对磁场区域肿瘤细胞抑制增殖作用强。细胞内的T2场核磁信号强度随C60-IONP-PEG浓度的增加而减弱，其核磁成像的图像呈现明显的黑化效应。在体内光疗活性评价中，以接种B16-F10细胞的C57小鼠为模型动物，分别考察了C60-IONP-PEG/HMME、C60-IONP-PEG、HMME在光照和不光照条件下对小鼠黑色素瘤抑制作用，结果表明C60-IONP-PEG/HMME在磁场作用下可滞留于肿瘤部位，532nm激光照射后发挥高效的光疗作用，其效果明显优于单纯的C60或HMME。此外，C60-IONP-PEG能够使荷瘤小鼠肿瘤、肝、脾、肾等器官的T2场信号显著降低，具有良好的负性增强效果，可作为体内核磁成像T2场的造影剂。以上结果表明，多功能肿瘤靶向光疗系统C60-IONP-PEG/HMME具有良好的磁靶向性、高效的光疗活性和卓越的核磁成像能力，在肿瘤诊断和治疗中具有潜在的应用价值。
[Abstract]:Fullerene (C60), with its unique structure and special optical properties, has potential application value in drug transport and photodynamic therapy, but its insolubility greatly restricts its application in the field of biomedicine. The purpose of this study is to modify the fullerenes and connect magnetic iron oxide to the surface of the fullerenes. Rice grain (IONPs) and hydrophilic group polyethylene glycol 2000 (PEG2000) have developed a new magnetic water soluble drug transport carrier, C60-IONP-PEG, and carry small molecular photosensitizer, porphyrin monomethyl ether (HMME), to construct a multi-functional tumor targeting drug delivery system with good magnetic targeting ability, phototherapy ability and nuclear magnetic imaging ability, C60-IONP-PEG/HMME.
First, a drug carrier based on fullerene derivatives was constructed. The Bingle ring addition reaction, ester hydrolysis reaction, thermochemical bonding reaction were used to synthesize the fullerene (C60-IONP-COOH) loaded with iron oxide, and the magnetic C60 load was prepared by the addition of the amino group of 2000 (PEG (2000) -NH2) with the carboxyl group of the fullerene derivative. The body system C60-IONP-PEG. uses Fourier transform infrared spectroscopy to verify the success of the carrier system, using transmission electron microscope, laser nanoscale analyzer and so on for morphological characterization, using a vibrating sample magnetometer to detect the magnetic properties, using a thermogravimetric analyzer to determine the content of PEG in the system, and using 3T NMR scanner. The results show that the magnetic carrier system is successfully prepared. C60-IONP-PEG has high water solubility, obvious superparamagnetic and significant magnetic imaging ability, and has a suitable particle size and potential. It can be used as a good carrier for magnetic targeting drug delivery.
In the preparation of magnetic targeting nanoscale preparation, using the small molecular photosensitizer HMME as the model drug, the influence of the dosage of HMME and the carrier material, the ultrasonic time and the super power on the dosage of drug delivery was investigated by single factor experiment, and the best prescription technology for the construction of the tumor targeting drug delivery system C60-IONP-PEG/HMME was determined. Transmission electron was used. The system was characterized by microscope, laser nanoscale analyzer and UV full wavelength scanner. The results showed that C60-IONP-PEG could efficiently load HMME (~125.3%). The system had good stability and uniform size, sustained release drug and sustained release of drugs for 120 hours.
In the evaluation of anti tumor activity in vitro, the mouse source melanoma (B16-F10) cells were selected as the research object, and the synergistic effect of HMME and C60 on the photodynamic treatment of the tumor was studied. The results showed that the blank carrier C60-IONP-PEG had no obvious toxicity to B16-F10 cells, but it could produce the living oxygen efficiently under the 532nm laser irradiation. The photosensitizer is used in photodynamic therapy. In addition, after 532nm laser irradiation, both C60 and HMME in the C60-IONP-PEG/HMME system produce the living oxygen efficiently, synergistically enhance the phototherapy effect, kill the tumor cells together, and have the concentration and light intensity dependence. The experimental results also show that C60-IONP-PEG/HMME has good magnetic targeting and magnetic field. Regional tumor cells have a strong inhibitory effect on proliferation. The intensity of nuclear magnetic signal in T2 field decreases with the increase of C60-IONP-PEG concentration, and the image of NMR imaging shows obvious blackening effect.
In the evaluation of phototherapy activity in vivo, C57 mice inoculated with B16-F10 cells were used as model animals. The inhibitory effects of C60-IONP-PEG/HMME, C60-IONP-PEG and HMME on the melanoma of mice under light and light conditions were investigated. The results showed that C60-IONP-PEG/HMME could be retained at the tumor site under the effect of magnetic field, and 532nm laser irradiated high efficiency. The effect of phototherapy is obviously better than that of simple C60 or HMME.. C60-IONP-PEG can significantly reduce the T2 field signal of tumor, liver, spleen and kidney in tumor bearing mice. It has a good negative enhancement effect and can be used as a contrast agent in the T2 field of MRI in vivo.
The above results show that the multi-functional tumor targeted phototherapy system C60-IONP-PEG/HMME has good magnetic targeting, high effective phototherapy activity and excellent magnetic imaging ability, and has potential application value in the diagnosis and treatment of tumor.

【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

【参考文献】