【作者】 孙军；

【导师】 高峰；

【作者基本信息】 华东理工大学 ， 药剂学， 2011， 硕士

【摘要】 他克莫司(Tacrolimus, FK506)是一种强效的免疫抑制剂,因在水中溶解度低限制了其应用。本课题通过采用超声法制备了FK506-二甲基-β-环糊精(DM-β-CD)包合物,采用去溶剂化-化学交联法制备了牛血清白蛋白(BSA)纳米粒(NP),最终制得FK506-DM-β-CD-BSA NP剂型,为新型纳米给药系统提供了实验数据。采用研磨法或超声法制备FK506-DM-β-CD包合物,结果发现超声法优于研磨法。采用超声法,通过正交试验,最优化包合物备工艺条件为：超声时间0.5 h,温度40℃,乙醇浓度75%,制得的FK506-DM-β-CD包合物的载药率为(12.4±0.3)%,极大提高了FK506的溶解度。采用去溶剂化-化学交联法制备BSA NP。通过单因素试验法考察交联时间、pH、白蛋白浓度、乙醇用量等因素对纳米粒形态、粒径、产率、体外降解过程的影响。结果发现控制乙醇用量可得到球形良好、140-300nm的多尺度纳米粒。按白蛋白浓度50mg/ml,pH9,乙醇用量4 ml,交联18h制得的纳米粒呈球形,平均粒径为(148.4±7.9)nm,产率为(97.5±0.1)%,15 min以内于人工胃液中降解(81.9±0.4)%,24 h以内于人工肠液中降解(18.8±0.1)%。添加冻干保护剂,通过冷冻干燥制得白蛋白纳米剂型。采用去溶剂化-化学交联法制备FK506-DM-β-CD-白蛋白纳米粒。当FK506浓度为3.5mg/ml时,FK506-DM-β-CD-BSA NP的包封率和载药率分别为(91.5±1.0)%和(3.2±0.1)%,粒径为(245.2±7.5)nm；与FK506-BSA NP比较, FK506-DM-β-CD-BSA NP的载药率从2.1%提高至3.2%,酶降解速度减慢,体外累积释放表明其突释更小,释药缓慢；药动学实验结果表明,大鼠静脉注射FK506-DM-β-CD-BSA NP后的血药浓度平稳且持久,其绝对生物利用度为(117.6±3.5)%。更多还原

【Abstract】 Tacrolimus (FK506) is a potent immunosuppressant, but its poor solubility restricted its application. FK506-dimethyl-(3-cyclodextrin (DM-β-CD) inclusion compound was prepared by an ultrasonic method. Bovine serum albumin (BSA) nanoparticle (NP) was prepared by a desolvation-chemical crosslinking method. FK506-DM-β-CD-BSA NP was prepared and these experimental data for a novel drug delivery system were provided.FK506-DM-P-CD inclusion compound was prepared by grind method or ultrasonic method and the results indicated that ultrasonic method was better than grind method. Then, the inclusion compound formation condition optimized by orthogonal test was 0.5 h,40℃and 75% ethanol under ultrasound. The loading efficiency of inclusion compound prepared by the above conditions was (12.4±0.3)%, while the inclusion compound greatly improved the solubility of FK506.BSA NP was prepared using a desolvation-chemical crosslinking method. The effects of crosslinking time, pH, albumin concentration, and the volume of ethanol were investigated by a single factor test. The preparations were optimized with the morphology, particle size, and yield as the indexes. The degradation of albumin nanoparticles was evaluated in vitro. These results found that the controllable nanopartieles between 140 and 300 nm were prepared by altering ethanol volume. The NP was spherical with an average diameter of (148.4±7.9)nm, yield of (97.5±0.1)% with the preparation conditions as follows:albumin 50 mg/ml, pH 9, ethanol 4 ml, and 18 h for crosslinking. Degradation of the BSA NP was (81.9±0.4)% in artificial gastric juice after 15 min and (18.8±0.1)% in artificial intestinal juice after 24 h, respectively. Albumin nanopreparation was obtained with protective agent after freeze-drying.FK506-DM-P-CD-BSA NP was also prepared by a desolvation-chemical crosslinking method. When FK506 was chosen at 3.5 mg/ml, the entrapment efficiency and loading efficiency of FK506-DM-β-CD-BSA NP were (91.5±1.0)% and (3.20±0.04)%, respectively, its particle size was (245.2±7.5)nm. Comparing with FK506-BSA NP, FK506-DM-β-CD-BSA NP increased by 2.12% to 3.20% in loading efficiency, and decreased by degradation in enzyme. The cumulative release curves indicated that FK506-DM-β-CD-BSA NP had lower break-release and slower drug release. These results of pharmacokinetical study showed that it maintained stable blood drug level and held time longer for intravenous injection of FK506-DM-β-CD-BSA NP in rats. The absolute bioavailability of FK506-DM-β-CD-BSA NP was (117.6±3.5)%.更多还原