多西他赛复合单壁碳纳米管固体脂质纳米粒的初步研究

发布时间：2019-01-03 17:22

【摘要】：多西他赛(DTX)是已上市的一种紫杉烷类抗癌药物，主要通过破坏肿瘤细胞的有丝分裂而达到抗肿瘤效果。目前多西他赛的上市剂型为注射液，其所含吐温80黏性较大且具有溶血作用，多数患者会产生明显的过敏反应。单壁碳纳米管(SWNT)具有独特大小与中空结构，因而具有独特的理化性质，并具有独特的跨膜能力，能够被动穿过多种细胞膜，通过内吞作用可携带多种活性分子进入细胞，因而在药物、核酸转运方面具有潜在的应用价值。但是未经处理的单壁碳纳米管（SWNT）的水溶性差，且具有一定的生物毒性，限制其在生物医学领域的应用。固体脂质纳米粒（SLN）具有生理相容性好，适用于多种给药形式、适合包载亲脂性药物、通过对其表面进行特征修饰，可将药物传递至特定组织，具有靶向性等优点，因此，SLN是一种具有发展前景的新型给药系统。本课题拟对单壁碳纳米管（SWNT）进行氧化，有效提高其水分散性；将多西他赛通过π-π堆积作用吸附到单壁碳纳米管的表面，制成固体脂质纳米粒(DTX/OSWNT/SLN)，使其具有被动靶向性；通过体外抗肿瘤活性作用的考察，开展抗癌药物肿瘤治疗的初步研究。本文首先对碳纳米管进行修饰，，使其水溶性得到一定程度的改善，且其明显被截短，更有利于载药和固体脂质纳米粒的构建。本文采用熔融超声法制备多西他赛复合单壁碳纳米管固体脂质纳米粒（DTX/OSWNT/SLN），并对其进行单因素考察和正交试验设计考察，最终确定其最佳制备工艺和最优处方。制备的DTX/OSWNT/SLN纳米粒呈较规则的球形或类球形，平均粒径为181.3±3.23nm，平均Zeta电位为-33.65±1.28mV；本文采用高效液相法测定制剂中多西他赛的含量，采用超滤-离心法测定包封率为96.43±0.97%，载药量为8.51±0.08%；选取海藻糖和蔗糖的混合物作为冻干保护剂，二者的总用量为5%（w/v）；体外释药试验结果表明原料药DTX在48h内释放完全，DTX/OSWNT/SLN表现出缓释作用，释药特征符合Weibull数学模型。以人源性乳腺癌细胞MCF-7细胞为受试细胞，考察制剂的体外抗肿瘤活性。试验结果表明，DTX/OSWNT/SLN可以携带药物转运到细胞内部；空白载体对细胞无明显的增殖抑制作用；制剂组和原料组对细胞的抑制率呈浓度依赖性和时间依赖性，而且DTX/OSWNT/SLN组对细胞的抑制作用强于原料药DTX组，808nm激光照射有助于增强对肿瘤细胞的抑制作用；原料组和制剂组对MCF-7细胞周期抑制均显著地反应在G2/M期，且均可诱导MCF-7细胞的凋亡。综上所述，本文成功制备了多西他赛复合单壁碳纳米管固体脂质纳米载体DTX/OSWNT/SLN，并对其理化性质及体外抗肿瘤活性进行考察，初步认为该纳米载体是一种很有潜力的抗肿瘤药物转运系统。
[Abstract]:Docetaxel (DTX) (docetaxel) is a taxane anticancer drug, which is mainly used to destroy the mitosis of tumor cells. The dosage form of docetaxel is injection, its Tween80 is viscous and hemolytic, and most patients have obvious allergic reaction. Single walled carbon nanotubes (SWNT) have unique size and hollow structure, so they have unique physical and chemical properties and unique transmembrane capability. They can passively penetrate a variety of cell membranes and carry a variety of active molecules into cells through endocytosis. Therefore, it has potential application value in drug and nucleic acid transport. However, untreated single-walled carbon nanotubes (SWNT) have poor water solubility and biological toxicity, which limits their application in biomedical field. Solid lipid nanoparticles (SLN) have good physiological compatibility, suitable for various forms of drug delivery, suitable for encapsulating lipophilic drugs. By modifying their surface characteristics, they can be transferred to specific tissues and have the advantages of targeting. SLN is a new drug delivery system with development prospects. In order to improve the water dispersion of single-walled carbon nanotubes (SCNTs), the (SWNT) was oxidized in this paper. The docetaxel was adsorbed on the surface of single-walled carbon nanotubes by 蟺-蟺 accumulation, and the solid lipid nanoparticles (DTX/OSWNT/SLN) were prepared, which made them passive targeting. The primary study of anticancer drug tumor therapy was carried out through the investigation of anti-tumor activity in vitro. In this paper, carbon nanotubes (CNTs) were modified to some extent to improve their water solubility, and they were obviously truncated, which was more favorable to the construction of drug loading and solid lipid nanoparticles. In this paper, docetaxel composite single-walled carbon nanotube solid lipid nanoparticles (DTX/OSWNT/SLN) were prepared by melt ultrasonic method, and were investigated by single factor and orthogonal design. Finally, the optimum preparation process and the optimal formulation were determined. The prepared DTX/OSWNT/SLN nanoparticles were spherical or spherical in shape, with an average diameter of 181.3 卤3.23 nm and an average Zeta potential of -33.65 卤1.28 MV. In this paper, the content of docetaxel in the preparation was determined by HPLC, the entrapment efficiency was 96.43 卤0.97 by ultrafiltration centrifugation, and the drug loading was 8.51 卤0.08. The mixture of trehalose and sucrose was chosen as freeze-drying protectant, the total dosage of them was 5% (w / v). The results of in vitro drug release test showed that DTX was completely released within 48 hours and DTX/OSWNT/SLN showed a slow release effect. The drug release characteristics were in accordance with the Weibull mathematical model. The anti-tumor activity of human breast cancer cell line MCF-7 was investigated in vitro. The results showed that DTX/OSWNT/SLN could carry drugs into the cells, and the blank vector had no obvious inhibitory effect on cell proliferation. The inhibition rate of cells in preparation group and raw material group was concentration-dependent and time-dependent, and the inhibitory effect of DTX/OSWNT/SLN group was stronger than that of DTX group. 808nm laser irradiation could enhance the inhibitory effect on tumor cells. The inhibition of MCF-7 cell cycle in raw material group and preparation group was significantly inhibited in G _ 2 / M phase and could induce apoptosis of MCF-7 cells. To sum up, docetaxel composite solid lipid nanotubes (DTX/OSWNT/SLN,) was successfully prepared and its physicochemical properties and antitumor activity in vitro were investigated. It is considered that this nano-carrier is a potential anti-tumor drug transport system.
【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;TB383.1

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