D-甘露糖修饰载光敏剂纳米体系的构建及其靶向光动力治疗乳腺癌的应用

发布时间：2018-05-25 02:19

本文选题：D-甘露糖 + BODIPY光敏剂　；参考：《江南大学》2017年硕士论文

【摘要】：传统的癌症(恶性肿瘤)治疗手段包括手术、化疗、放疗等,其中化疗是最主要的治疗手段之一。但由于化疗全身给药的特点,降低了抗癌药物的生物利用度,同时也会对正常组织和细胞产生毒副作用并引起癌细胞的耐药性等问题,这些严重限制了其在癌症治疗中的应用和疗效。通过不断研究与探索,科研工作者开发出各种靶向药物输送体系,在改善化疗性能方面成果显著。目前,一种新型安全高效的靶向光动力疗法被开发出来,并将其用于癌症治疗中,取得了很好的效果。它利用靶向输送载体,将低暗毒性的光敏药物富集于肿瘤部位,经特定波长的光照后,诱导产生氧化性很强的单线态氧,进而实现对癌细胞的杀伤。由于光敏药物良好的生物相容性和高效的光疗性能,这种用于光动力疗法的纳米光敏制剂在治疗恶性肿瘤的研究与应用中具有很大的潜力。本研究设计合成了一种共价键连金刚烷的三臂BODIPY光敏剂分子,通过β-环糊精和金刚烷超分子识别作用将D-甘露糖化环糊精固定到光敏剂纳米粒子表面,从而制备了表面甘露糖分子功能修饰的光敏剂纳米粒子(BTA@CD-Man7)。这种设计赋予了BTA@CD-Man7的甘露糖受体靶向性和光动力治疗的功能。CD-Man7和BTA的分子结构通过核磁共振氢谱和时间辅助飞行质谱等进行确认。BTA@CD-Man7的形貌和流体力学直径及溶液稳定性通过透射电镜和动态光散射进行考察,结果表明,甘露糖对于维持生物溶剂中BTA@CD-Man7纳米结构的均一稳定方面具有重要作用。通过荧光分光光度计测定单线态氧捕获剂ABDA荧光强度的变化来表征不同状态下BTA@CD-Man7产单线态氧的性能,结果表明BTA@CD-Man7需解组装释放光敏剂BTA,并在665 nm LED光源辐射下,才能产生单线态氧。以表面甘露糖受体过度表达的人乳腺癌MDA-MB-231细胞作为模型,通过激光共聚焦显微镜评价BTA@CD-Man7经受体识别介导的细胞内吞作用和光动力治疗性能。实验结果表明表面甘露糖受体高表达的癌细胞MDA-MB-231能够大量摄取BTA@CD-Man7,并在溶酶体中解组装释放光敏剂BTA,在665 nm LED灯光照下生成大量的单线态氧,单态氧能与附近的生物大分子发生氧化反应,产生细胞毒性进而导致癌细胞凋亡。而对于表面甘露糖受体低表达的正常细胞MCF-10A,BTA@CD-Man7则很少被摄取,且光动力治疗效率低。最后,采用MTT细胞毒性评价法评价BTA@CD-Man7和BTA@CD的细胞光暗毒性。结果表明BTA@CD-Man7和BTA@CD具有良好的生物相容性,在黑暗条件下,两者对癌细胞与正常细胞几乎没有细胞毒性,而在给予665 nm LED光照的条件下,BTA@CD-Man7表现出更加高效的靶向光疗性能。荷瘤(MDA-MB231)小鼠光疗实验进一步证明BTA@CD-Man7在靶向光动力治疗乳腺癌的应用中很有潜力。
[Abstract]:The traditional treatment of cancer (malignant tumor) includes surgery, chemotherapy, radiotherapy and so on, among which chemotherapy is one of the most important treatment methods. However, because of the characteristics of systemic chemotherapy, the bioavailability of anticancer drugs is reduced, and at the same time, toxic side effects on normal tissues and cells and drug resistance of cancer cells are also caused. These severely limit its application and efficacy in cancer treatment. Through continuous research and exploration, researchers have developed various targeted drug delivery systems, which have achieved remarkable results in improving the performance of chemotherapy. At present, a new type of safe and efficient targeted photodynamic therapy has been developed and used in cancer treatment, and good results have been obtained. It uses the targeted transport carrier to enrich the low dark toxic Guang Min drugs in the tumor site, and after a specific wavelength of light, induces the production of highly oxidizing singlet oxygen, and then realizes the killing of cancer cells. Because of the good biocompatibility and high phototherapy performance of Guang Min drugs, this kind of nanometer Guang Min preparation used in photodynamic therapy has great potential in the research and application of malignant tumor treatment. In this study, a three-arm BODIPY Guang Min molecule with covalent bond and adamantane was designed and synthesized. D-mannose cyclodextrin was immobilized on the surface of Guang Min nanoparticles by 尾 -cyclodextrin and adamantane supramolecular recognition. Therefore, Guang Min nanoparticles modified by mannose molecular function were prepared. This design gives BTA@CD-Man7 the function of mannose receptor targeting and photodynamic therapy. CD-Man7 and BTA molecular structures are confirmed by NMR and time-assisted flight mass spectrometry. The morphology and hydrodynamic diameter and solution of BTA CD-Man7 are confirmed by nuclear magnetic resonance spectroscopy (NMR) and time-assisted flight mass spectrometry (TFAMS). The stability was investigated by transmission electron microscopy (TEM) and dynamic light scattering (DLS). The results show that mannose plays an important role in maintaining the homogeneity and stability of BTA@CD-Man7 nanostructures in biological solvents. The change of fluorescence intensity of single line oxygen trapping agent (ABDA) was measured by fluorescence spectrophotometer to characterize the properties of single line oxygen produced by BTA@CD-Man7 in different states. The results showed that BTA@CD-Man7 needed to be unassembled and released Guang Min, which was irradiated by LED light source at 665nm. In order to produce singlet oxygen. Human breast cancer MDA-MB-231 cells with overexpression of mannose receptor on the surface were used as a model to evaluate the intracellular endocytosis and photodynamic therapy of BTA@CD-Man7 mediated by receptor recognition by confocal laser microscopy. The experimental results showed that MDA-MB-231, which was highly expressed on the surface of mannose receptor, was able to ingest BTA-CD-Man7 in large quantities, and unassemble and release Guang Min in lysosome, resulting in a large amount of singlet oxygen under the irradiation of 665nm LED light. Single state oxygen can oxidize the nearby biomolecules, resulting in cytotoxicity and apoptosis of cancer cells. However, MCF-10AnBTACD-Man7 cells with low surface mannose receptor expression were rarely ingested, and the photodynamic therapy efficiency was low. Finally, the cytotoxicity of BTA@CD-Man7 and BTA@CD was evaluated by MTT cytotoxicity evaluation method. The results showed that BTA@CD-Man7 and BTA@CD had good biocompatibility. Under dark conditions, BTA@CD-Man7 and BTA@CD had little cytotoxicity to cancer cells and normal cells, while CD-Man7 showed a more efficient phototherapeutic performance under the irradiation of 665nm LED. The phototherapy of mice with MDA-MB231 further demonstrated the potential of BTA@CD-Man7 in the application of targeted photodynamic therapy for breast cancer.
【学位授予单位】：江南大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R730.5;TB383.1

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