紫杉醇纳米脂质体凝胶剂的制备与评价

发布时间：2018-05-27 03:09

本文选题：紫杉醇 + 纳米脂质体　；参考：《浙江大学》2014年硕士论文

【摘要】：目的：探讨紫杉醇纳米脂质体凝胶剂的制备方法、体外评价及其对大鼠角叉菜胶致足肿胀模型及小鼠甲醛致痛模型的镇痛抗炎作用,以期为紫杉醇开发成治疗类风湿性关节炎的新制剂奠定实验基础。方法：1、建立紫杉醇的HPLC色谱分析法,以及紫杉醇纳米脂质体包封率的测定方法。2、采用薄膜分散高压微射流法制备紫杉醇纳米脂质体,冷冻干燥至粉末状,以包封率、粒径为指标采用正交试验筛选最佳工艺。采用透射电镜和纳米粒径测定仪观测其形态,粒径大小及分布。3、以粘度、基质色泽、涂展性及均匀度为观察指标,确定紫杉醇纳米脂质体凝胶剂最佳基质,以累积释放量为指标采用正交试验筛选最佳工艺。制备紫杉醇纳米脂质体凝胶剂并考察其体外稳定性。采用透析膜扩散法进行体外释放试验,以离体小鼠皮结合改良Franz扩散装置考察其体外透皮特性以及药物在小鼠皮肤中的分布情况。4、进一步考察紫杉醇纳米脂质体凝胶剂的皮肤刺激性、过敏反应及对小鼠甲醛致痛模型及大鼠角叉菜胶致足肿胀模型的镇痛抗炎作用。结果：1、建立了简单可行,操作简便的HPLC色谱法,用于测定紫杉醇。确定了紫杉醇纳米脂质体包封率的测定方法。2、确定紫杉醇纳米脂质体的最佳工艺为：卵磷脂含量为2%,药物与磷脂质量比为1：30,磷脂与胆固醇的质量比为10：1。采用最佳处方工艺制得的脂质体粒径为81.8nm；粒径分布系数为0.18；平均包封率73.2%。通过透射电镜观察,纳米脂质体圆形或类球形,分散性良好,具明显指纹状结构。3、通过筛选确定凝胶基质为卡波姆,紫杉醇纳米脂质体凝胶剂的最佳处方：卡波姆3%,甘油5%,透皮渗透剂3%氮酮。72h药物累积释放百分率为79.04%；48h的药物单位面积累积渗透量为429.68μg.cm-2,与紫杉醇凝胶剂比较,脂质体能显著促进紫杉醇透过皮肤(P0.05)。紫杉醇纳米脂质体给药4h后在角质层和活性皮肤层中的药物含量分别为3.41μg.cm-2.5.35μg.cm-2,与紫杉醇凝胶剂比较,能显著提高药物在皮肤中的滞留量(P0.05)。4、紫杉醇纳米脂质体用于皮肤后无明显的刺激作用,与豚鼠皮肤反复接触后未见过敏反应的发生。抗炎实验中,与模型组比较,各给药组在一定时间内均能显著减轻致炎足爪的肿胀程度(P0.05),并呈现明显的量效关系。甲醛致痛实验中,给药组与模型组比较,小鼠舔足次数明显减少(P0.05)。结论：该制剂制备工艺简单,易于涂布,粒径较小且分布均匀,体外释放缓慢,具有明显的缓释作用,与紫杉醇凝胶剂相比,纳米脂质体能增加药物的皮肤滞留量,促进脂溶性药物紫杉醇透过皮肤并具有明显镇痛抗炎作用。
[Abstract]:Objective: to investigate the preparation and in vitro evaluation of paclitaxel nano-liposome gel and its analgesic and anti-inflammatory effects on rat paw swelling induced by carrageenin and formaldehyde-induced pain in mice. In order to lay the experimental foundation for taxol to develop into a new preparation for the treatment of rheumatoid arthritis. Methods HPLC chromatographic analysis of paclitaxel and determination of encapsulation efficiency of paclitaxel nanoliposomes were established. Paclitaxel nanoliposomes were prepared by thin-film dispersion high-pressure micro-jet method. The best technology was selected by orthogonal test. The morphology, size and distribution of paclitaxel nano-liposomes were observed by transmission electron microscope (TEM) and nano-particle size analyzer. The optimum matrix of paclitaxel nano-liposome gel was determined by using viscosity, color and color of matrix, spreading property and uniformity as observation indexes. The best technology was screened by orthogonal experiment with cumulative release amount as index. Paclitaxel nano-liposome gel was prepared and its stability in vitro was investigated. In vitro release test was carried out by dialysate membrane diffusion method. The in vitro transdermal properties and drug distribution in mouse skin were investigated by using an isolated mouse skin and modified Franz diffusion apparatus. The skin irritation of paclitaxel nano-liposome gel was further investigated. Anaphylaxis and its analgesic and anti-inflammatory effects on formaldehyde-induced pain model and rat paw swelling model induced by carrageenin. Results: a simple and convenient HPLC chromatography was established for the determination of paclitaxel. The entrapment efficiency of paclitaxel nanoliposomes was determined as follows: the content of lecithin was 2, the mass ratio of drug to phospholipid was 1: 30, and the mass ratio of phospholipid to cholesterol was 10: 1. The particle size of liposome was 81.8 nm, the particle size distribution coefficient was 0.18, and the average entrapment efficiency was 73.2%. The nano-liposomes were round or globular by transmission electron microscope, and had good dispersibility and obvious fingerprint structure. The gel matrix was determined to be carbomer by screening. The best formulation of paclitaxel nano-liposome gel: carbomer 3um, glycerol 5h, transdermal osmotic agent 3% azone .72h drug cumulative release percentage 79.04g 路cm ~ (-2) per unit area of the drug was 429.68 渭 g 路cm ~ (-2), compared with paclitaxel gel. Lipid can significantly promote paclitaxel through the skin P 0.05. The contents of paclitaxel nanoliposomes in corneum and active skin layer were 3.41 渭 g.cm-2.5.35 渭 g 路cm-2, respectively, which were compared with paclitaxel gel. Paclitaxel nanoliposomes had no obvious stimulative effect on skin, and no allergic reaction occurred after repeated contact with guinea pig skin. In the anti-inflammatory experiment, compared with the model group, each group could significantly reduce the swelling degree of paw in a certain time, and showed a significant dose-effect relationship. In the experiment of formaldehyde-induced pain, compared with the model group, the mice licked their feet significantly. Conclusion: compared with paclitaxel gel, nano-liposomes can increase the skin retention of the drug, with simple preparation process, easy coating, small particle size and uniform distribution, slow release in vitro and obvious slow-release effect. Promote fat-soluble drug paclitaxel through the skin and have obvious analgesic anti-inflammatory effect.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R943

【参考文献】