他克莫司—二甲基-β-环糊精包合物及白蛋白纳米剂型的研究
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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)%。
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)%.
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)%。
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)%.
引文
[1]王永,陆兵,朱厚础.纳米粒作为脑靶向给药系统的研究进展[J].解放军药学学报,2006,22(3)：202-205.
[2]张阳德.纳米药物学[M].北京：化学工业出版社,2006.193.
[3]王朝辉,姜同英,张婧.脑靶向纳米载体系统的研究进展[J].中国生化药物杂志,2008,29(5),355-358.
[4]Srinivas G, Harikrishna D, Aliasgar S, et al.A. review of stimuli-responsivenano carrier for drug and gene delivery[J]. J Controlled Release,2008,126:187-204.
[5]陈桐楷,李园,林华庆.固体脂质纳米粒给药系统新载体的研究进展[J].中国医药导报2009,24：10-12.
[6]张丽碚,方晓玲.聚合物纳米粒和胶束的主动靶向研究进展[J].国外医学：药学分册,2005,32(6)：399-402.
[7]刘惠娟,李平.磁性载体在药物传递系统中的应用[J].中国药师,2009,5：663-666.
[8]吴珍,杜士明,董永成.纳米粒作为脑靶向给药系统的研究进展[J].中国药房,2007,18(13)：1029-1031.
[9]S Dreis, F Rothweiler, M Michaelis, et al. Preparation, characterisation and maintenance of drug efficacy of doxorubicin-loaded human serum albumin (HSA) nanoparticles[J]. Int J Pharm,2007,341:207-214.
[10]Marion G A, Hanns C M, Klaus L. Freeze drying of human serum albumin (HSA) nanoparticles with different excipients[J]. Int J Pharm,2008,363:162-169.
[11]胡连栋,唐星,崔福德.维甲酸固体脂质纳米粒冷冻干燥工艺研究[J].中国医药工业杂志,2007,38(1)：22-26.
[12]吴镭,平其能.药剂学发展与展望[M].北京：化学工业出版社,2002.63-64.
[13]王芬.纳米二氧化硅对PC12神经细胞及HEK293人胚肾细胞毒性的研究[M].上海：华东理工大学,2009.
[14]Storm G, Belliot S O, Daemen T. Surface modification of nanoparticles to oppose uptake by the mononuclear phagocyte system[J]. Adv Drug Deliv Rev,1995,16:190-197.
[15]Lin W, Garnett M C, Schacht E, et al. Preparation and in vitro characterization of HSA-mPEG nanoparticles[J]. Int J Pharm,1999,189 (2):161-170.
[16]张良珂,侯世祥,毛声俊.受体介导米托蒽醌白蛋白纳米粒肿瘤细胞靶向性研究[J]. 四川大学学报(医学版),2006,37(1)：77-79.
[17]Wartllck H, Schmitt S, Strebhardt K, et al. Tumor cell delivery of antisense oligonucleotidos by human serum albumin nanoparticles[J]. J Controlled Release,2004,96
(3):483-495.
[18]齐崴,何志敏.牛血清白蛋白的同源模建与结构解析[J].计算机与应用化学,2006,23(1)：821-824.
[19]苏华,胡晋红,李凤前.白蛋白纳米粒的制备工艺及其靶向性研究进展[J].中国药学杂志,2005,40(9)：641-644.
[20]杨莹.白蛋白纳米粒的制备工艺及在药学中的应用[J].今日药学,2009,19(4)：57-59.
[21]Muller B G, Leuenberger H, Kissel T. Albumin Nanospheres as Carrier for Passive Drug Targeting:An Optimized Manufacturing Technique [J]. Pharm Res,1996,13 (1):32-37.
[22]石莉,邓意辉,董晓辉等.超声法制备齐多夫定碳酸胆固醇酯牛血清白蛋白纳米粒[J].沈阳药科大学学报,2008,25(10)：759-764.
[23]Weber C, Coester C, Kreuter J, et al. Desolvafion process and Surface characteristics of protein nanoparticles[J]. Int J Pharm,2000,196:197-200.
[24]K Langer, S Balthasar, V Vogel, et al. Optimization of the preparation process for human serum albumin (HSA) nanoparticles[J]. Int J Phann,2003,257:169-180.
[25]Vogel V, Langer K, Balthasar S, et al. Characterization of serum albumin nanoparticles by sedimentation velocity analysis and electron microscopy[J]. Prog Colloid Polym Sci,2002,119:28-33.
[26]Coombes A G, Breeze V, Wu Lin, et al. Lactic acid-stabilised albumin for mierosphere formulation and biomedical coatings[J]. Biomaterials,2001,22:1-5.
[27]Chen C O, Lin W, Coombes A G, et al. Preparation of human serum albumin microspheres by a novel acetone-heat denaturation method[J]. J Microencapsulation,1994, 11(4),395-407.
[28]Merodio M, Irache J M, Eclancher F, et al. Ganciclovir-loaded albumin nanoparticles: characterization and in vitro release proterties[J]. Eur J Pharm Biopharm,2001,12(3):249-253.
[29]孙望强.大分子纳米粒的制备工艺及其性能研究[D].武汉：华中科技大学,2004.
[30]何治柯,梁建功,谢海燕等.水溶性纳米粒子的制备方法.专利公开号CN 1431070A.
[31]Meziani M J, Sun Y P. Prntein-conjugated nanourtieles from rapid expansion of supereritieal fluid lufion into aqueous solution[J]. J Am Chem Soc,2003,125 (26):8015-8018.
[32]Hyuncheol K, Shaun B R, Karl G C. Investigating the Movement of Intravitreal Human Serum Albumin Nanoparticles in the Vitreous and Retina[J]. Pharm Res,2009,26 (2)：329-337.
[33]张良珂,侯世祥,宋相容等.一种白蛋白纳米粒的制备与评价[J].中国药学杂志,2007,42(5)：365-367.
[34]郭莉媛,姚晶萍,隋丽华.白藜芦醇白蛋白纳米粒的制备及其抗卵巢癌细胞增殖作用的研究[J].高等学校化学学报,2009,30：474-477.
[35]张晓燕,平其能.多西紫杉醇白蛋白纳米粒的制备及体外评价[J].药学进展,2008,32(5)：223-228.
[36]王恺,马光辉.白蛋白纳米球药物载体的制备及表征[J].过程工程学报,2004,4(2)：155-159.
[37]张良珂,侯世祥,卢懿等.叶酸偶联米托蒽醌白蛋白纳米粒的制备及体外性质研究[J].中国药学杂志,2005,40(10)：763-765.
[38]Guilin W, Kevin Siggers, Sufeng Zhang, et al. Preparation of BMP-2 Containing Bovine Serum Albumin (BSA) Nanoparticles Stabilized by Polymer Coating[J]. Pharm Res, 2008,25 (12):2896-2909.
[39]林川.他克莫司的药理作用及临床应用概述[J].海峡药学,2007,19(9)：87-89.
[40]Petrers, P H, Fittona, et al. Tacrolimus:A review of pharmacology and therapeutic potential in hepatic and renal transplantation[J]. Drugs,1993,46(4):746-749.
[41]Hutehinson I V. The mode of action of prograf (FK506) and its significance for long term graft survive. New Horizons in Kidney[J]. Transplantation,1997,1:22-26.
[42]Yoshihiro M, Hiroyuki T, Akihiro N, et al. PET measurement of FK506 concentration in a monkey model of stroke[J]. Nucl. Med Biol,2007,34:703-707.
[43]Alf L, Hiromitsu Y, Hirofumi T, et al. A pH-sensitive microsphere system for the colon delivery of tacrolimus containing nanoparticles[J]. Pharmacol Exp Ther,2005,104:337-346.
[44]Alf L, Hiromitsu Y, Hirofumi T, et al. Nanoparticles Enhance Therapeutic Efficiency by Selectively Increased Local Drug Dose in Experimental Colitis in Rats[J]. J Pharmacol Exp Ther,2005,315:196-202.
[45]费文雷,陈家祺,庞志清等.FK506纳米粒制备及在兔眼组织中的分布[J].眼科研究,2004,22(2)：164-166.
[46]费文雷,陈家祺,钟诗龙等.FKS06及其纳米粒的房水药代动力学的实验研究[J].中华眼科杂志,2006,42(4)：305-308.
[47]王启平,李东,张振伟等.他克莫司纳米微球的制备及其体外释药性能[J].中国现代药物应用,2008,2(14)：21-23.
[48]武国军,文爱东等.普乐可复在肾移植患者体内的药代动力学研究探讨[J].第四军医大学学报,2001,22(20)：1909-1911.
[49]Mekki O, Lee C, Carrier GJ. The effect of food on oral bioavailability of tacrolimus (FK506) in liver transplant patients[J]. Abstract Clin Pharmcol Ther,1993,5(2):229-230.
[50]钟秀英.药用p-环糊精包合物制备、检验技术研究进展[J].中药材,2003,26(4)：301-305.
[51]Hassan N, Asghar K. Investigation dchanism of β-lactam conjugated telechelic polymers of PEG and β-cyclodextrin as the new nanosized drug carrier devices[J]. Carbo Polym,2009, 76 (1) 46-50.
[52]谷福根,高永良,崔福德.环糊精包合物研究进展[J].中国新药杂志,2005,14(6)：686-693.
[53]张莉,罗来辉.环糊精包合物的研究进展[J].华北工学院学报,2003,24(4)：278-281.
[54]Thompson D O. Cyclodextrins enabling excipients:their present and future use in pharmaceuticals[J]. Crit Rev Ther Drug Carrier Syst,1997,14 (1) 101-104.
[55]Li J, Loh X J. Cyclodextrin-based supramolecular architectures:Syntheses, structures, and applications for drug and gene delivery[J]. Adv Drug Delivery Rev,2008,60 (9): 1000-1017.
[56]马开,秦文杰,高寒.痛可停胶囊挥发油p-环糊精包合物的实验研究[J].中药新药与临床药理,2003,14(6)：409-413.
[57]丁燕飞,姚瑶,周彦彬等.尼群地平p-环糊精包合物的研制[J].中国医院药学杂志,2003,23(8)：461-464.
[58]Stella VJ, He Q. Cyclodextrins[J]. Toxicol Pathol,2008,36(1):30-42.
[59]尤思路,高秀蓉,廖昌军.饱和水溶液法制备葛根素的p-环糊精包合物研究[J].西南军医,2008,10(4)：17-19.
[60]易军,李晓芳,苏建湘等.超声法制备石菖蒲挥发油p-环糊精包合物[J].时珍国医国药,2006,17(3)：381-383.
[61]齐美玲,高晶,邵清龙.大蒜素-环糊精包合物的制备及大蒜素的含量测定[J].北京理工大学学报,2004,24(7)：650-655.
[62]邵伟,袁敏,王大庆等.黄芩甙-羟丙基-p-环糊精包合物的研究[J].中成药,2001,23(5)：318-322.
[63]孙伟张,曾仁杰,景莉等.喷雾干燥法制备盐酸异丙嗪p-环糊精包合物的实验研究[J].西南国防医药,2001,1](6)385-388.
[64]Beudad H, Legrand P, Lebas G, et al. Combined hydroxyl propyleyelodextrin and poly (alkylcyanoaerylatel nanoparticles intended for oral administration of saquinavir[J]. Int J Pharm,2001,218:113-124.
[65]Francesca M, Marcos G, Paola M, et al. A new drug nanocarrier consisting of chitosan and hydoxypropylcyclodextrin[J]. Eur J Pharm Biopharm,2006,63:79-86.
[66]Alexander H K, Mana J A. Chitosan/cyclodextrin nanoparticles as macromolecular drug delivery system[J]. Int J Pharm,2007,34:134-142.
[67]尹婕,邓盛齐.环糊精及其衍生物在新型给药系统中的应用进展[J].国外医药抗生素分册,2006,27(5)：228-232.
[68]Hidetoshi A, Kiyokazu Y, Kouzou M, et al. Comparative studies of the enhancing effects of cyclodextrins on the solubility and oral bioavailability of tacrolimus in rats[J]. J Pharm Sci,2001,90 (6):690-701.
[69]陶少林.灵芝酸单体Me肠溶胶囊及药动学的研究[M].上海：华东理工大学,2009.
[70]顾磊.两种交联剂在氟尿嘧啶壳聚糖微球制备中的研究[M].上海：华东理工大学,2008.
[71]K Langer, M G Anhorn, I Steinhauser, et al. Human serum albumin (HSA) nanoparticles:Reproducibility of preparation process and kinetics of enzymatic degradation[J]. Int J Pharm,2008,347:109-117.
[72]陆彬,李凤前.白蛋白微球的热变性对其水溶性及酶降解性的影响[J].药学学报,2000,35(7)：535-538.
[73]Feng Q L, Hua S, Jing W. Preparation and characterization of sodium ferulate entrapped bovine serum albumin nanoparticles for liver targeting[J]. Int J Pharm,2008,349:274-282.
[74]王旭波,徐希明,葛亮等.半乳糖化阿霉素白蛋白纳米粒的制备及其质量评价[J].中国新药杂志,2005,14(7)：870-872.
[75]苏华,胡晋红,李凤前.表面活性剂对高水溶性药物阿魏酸钠白蛋白纳米粒的载药特性影响[J].药学服务与研究,2005,5(2)：153-156.
[76]陈悦HPLC法测定他克莫司的含量及有关物质[J].药物分析志,2008,28(2)：309-313.
[77]郑国钢,管燕,罗金文.反相高效液相色谱法测定他克莫司胶囊的含量[J].中国药业,2007,16(1)：17-18.
[78]李国琴,曾苏.梯度洗脱高效液相色谱法检测他克莫司有关物质[J].中国药学杂志,2008,43(7)：551-558.
[79]Takeshi A, Tomoko N, Masaru Y, et al. Quantitative deterruination of tautonmric FK506 by reversed-phase liquid chromatography[J]. J Pharm Biomed Anal,1996,14:339-346.
[80]梁陈方,王亚洲.反相高效液相色谱法测定他克莫司的血药浓度[J].广西医学,2009,31(3)：425-426.
[2]张阳德.纳米药物学[M].北京：化学工业出版社,2006.193.
[3]王朝辉,姜同英,张婧.脑靶向纳米载体系统的研究进展[J].中国生化药物杂志,2008,29(5),355-358.
[4]Srinivas G, Harikrishna D, Aliasgar S, et al.A. review of stimuli-responsivenano carrier for drug and gene delivery[J]. J Controlled Release,2008,126:187-204.
[5]陈桐楷,李园,林华庆.固体脂质纳米粒给药系统新载体的研究进展[J].中国医药导报2009,24：10-12.
[6]张丽碚,方晓玲.聚合物纳米粒和胶束的主动靶向研究进展[J].国外医学：药学分册,2005,32(6)：399-402.
[7]刘惠娟,李平.磁性载体在药物传递系统中的应用[J].中国药师,2009,5：663-666.
[8]吴珍,杜士明,董永成.纳米粒作为脑靶向给药系统的研究进展[J].中国药房,2007,18(13)：1029-1031.
[9]S Dreis, F Rothweiler, M Michaelis, et al. Preparation, characterisation and maintenance of drug efficacy of doxorubicin-loaded human serum albumin (HSA) nanoparticles[J]. Int J Pharm,2007,341:207-214.
[10]Marion G A, Hanns C M, Klaus L. Freeze drying of human serum albumin (HSA) nanoparticles with different excipients[J]. Int J Pharm,2008,363:162-169.
[11]胡连栋,唐星,崔福德.维甲酸固体脂质纳米粒冷冻干燥工艺研究[J].中国医药工业杂志,2007,38(1)：22-26.
[12]吴镭,平其能.药剂学发展与展望[M].北京：化学工业出版社,2002.63-64.
[13]王芬.纳米二氧化硅对PC12神经细胞及HEK293人胚肾细胞毒性的研究[M].上海：华东理工大学,2009.
[14]Storm G, Belliot S O, Daemen T. Surface modification of nanoparticles to oppose uptake by the mononuclear phagocyte system[J]. Adv Drug Deliv Rev,1995,16:190-197.
[15]Lin W, Garnett M C, Schacht E, et al. Preparation and in vitro characterization of HSA-mPEG nanoparticles[J]. Int J Pharm,1999,189 (2):161-170.
[16]张良珂,侯世祥,毛声俊.受体介导米托蒽醌白蛋白纳米粒肿瘤细胞靶向性研究[J]. 四川大学学报(医学版),2006,37(1)：77-79.
[17]Wartllck H, Schmitt S, Strebhardt K, et al. Tumor cell delivery of antisense oligonucleotidos by human serum albumin nanoparticles[J]. J Controlled Release,2004,96
(3):483-495.
[18]齐崴,何志敏.牛血清白蛋白的同源模建与结构解析[J].计算机与应用化学,2006,23(1)：821-824.
[19]苏华,胡晋红,李凤前.白蛋白纳米粒的制备工艺及其靶向性研究进展[J].中国药学杂志,2005,40(9)：641-644.
[20]杨莹.白蛋白纳米粒的制备工艺及在药学中的应用[J].今日药学,2009,19(4)：57-59.
[21]Muller B G, Leuenberger H, Kissel T. Albumin Nanospheres as Carrier for Passive Drug Targeting:An Optimized Manufacturing Technique [J]. Pharm Res,1996,13 (1):32-37.
[22]石莉,邓意辉,董晓辉等.超声法制备齐多夫定碳酸胆固醇酯牛血清白蛋白纳米粒[J].沈阳药科大学学报,2008,25(10)：759-764.
[23]Weber C, Coester C, Kreuter J, et al. Desolvafion process and Surface characteristics of protein nanoparticles[J]. Int J Pharm,2000,196:197-200.
[24]K Langer, S Balthasar, V Vogel, et al. Optimization of the preparation process for human serum albumin (HSA) nanoparticles[J]. Int J Phann,2003,257:169-180.
[25]Vogel V, Langer K, Balthasar S, et al. Characterization of serum albumin nanoparticles by sedimentation velocity analysis and electron microscopy[J]. Prog Colloid Polym Sci,2002,119:28-33.
[26]Coombes A G, Breeze V, Wu Lin, et al. Lactic acid-stabilised albumin for mierosphere formulation and biomedical coatings[J]. Biomaterials,2001,22:1-5.
[27]Chen C O, Lin W, Coombes A G, et al. Preparation of human serum albumin microspheres by a novel acetone-heat denaturation method[J]. J Microencapsulation,1994, 11(4),395-407.
[28]Merodio M, Irache J M, Eclancher F, et al. Ganciclovir-loaded albumin nanoparticles: characterization and in vitro release proterties[J]. Eur J Pharm Biopharm,2001,12(3):249-253.
[29]孙望强.大分子纳米粒的制备工艺及其性能研究[D].武汉：华中科技大学,2004.
[30]何治柯,梁建功,谢海燕等.水溶性纳米粒子的制备方法.专利公开号CN 1431070A.
[31]Meziani M J, Sun Y P. Prntein-conjugated nanourtieles from rapid expansion of supereritieal fluid lufion into aqueous solution[J]. J Am Chem Soc,2003,125 (26):8015-8018.
[32]Hyuncheol K, Shaun B R, Karl G C. Investigating the Movement of Intravitreal Human Serum Albumin Nanoparticles in the Vitreous and Retina[J]. Pharm Res,2009,26 (2)：329-337.
[33]张良珂,侯世祥,宋相容等.一种白蛋白纳米粒的制备与评价[J].中国药学杂志,2007,42(5)：365-367.
[34]郭莉媛,姚晶萍,隋丽华.白藜芦醇白蛋白纳米粒的制备及其抗卵巢癌细胞增殖作用的研究[J].高等学校化学学报,2009,30：474-477.
[35]张晓燕,平其能.多西紫杉醇白蛋白纳米粒的制备及体外评价[J].药学进展,2008,32(5)：223-228.
[36]王恺,马光辉.白蛋白纳米球药物载体的制备及表征[J].过程工程学报,2004,4(2)：155-159.
[37]张良珂,侯世祥,卢懿等.叶酸偶联米托蒽醌白蛋白纳米粒的制备及体外性质研究[J].中国药学杂志,2005,40(10)：763-765.
[38]Guilin W, Kevin Siggers, Sufeng Zhang, et al. Preparation of BMP-2 Containing Bovine Serum Albumin (BSA) Nanoparticles Stabilized by Polymer Coating[J]. Pharm Res, 2008,25 (12):2896-2909.
[39]林川.他克莫司的药理作用及临床应用概述[J].海峡药学,2007,19(9)：87-89.
[40]Petrers, P H, Fittona, et al. Tacrolimus:A review of pharmacology and therapeutic potential in hepatic and renal transplantation[J]. Drugs,1993,46(4):746-749.
[41]Hutehinson I V. The mode of action of prograf (FK506) and its significance for long term graft survive. New Horizons in Kidney[J]. Transplantation,1997,1:22-26.
[42]Yoshihiro M, Hiroyuki T, Akihiro N, et al. PET measurement of FK506 concentration in a monkey model of stroke[J]. Nucl. Med Biol,2007,34:703-707.
[43]Alf L, Hiromitsu Y, Hirofumi T, et al. A pH-sensitive microsphere system for the colon delivery of tacrolimus containing nanoparticles[J]. Pharmacol Exp Ther,2005,104:337-346.
[44]Alf L, Hiromitsu Y, Hirofumi T, et al. Nanoparticles Enhance Therapeutic Efficiency by Selectively Increased Local Drug Dose in Experimental Colitis in Rats[J]. J Pharmacol Exp Ther,2005,315:196-202.
[45]费文雷,陈家祺,庞志清等.FK506纳米粒制备及在兔眼组织中的分布[J].眼科研究,2004,22(2)：164-166.
[46]费文雷,陈家祺,钟诗龙等.FKS06及其纳米粒的房水药代动力学的实验研究[J].中华眼科杂志,2006,42(4)：305-308.
[47]王启平,李东,张振伟等.他克莫司纳米微球的制备及其体外释药性能[J].中国现代药物应用,2008,2(14)：21-23.
[48]武国军,文爱东等.普乐可复在肾移植患者体内的药代动力学研究探讨[J].第四军医大学学报,2001,22(20)：1909-1911.
[49]Mekki O, Lee C, Carrier GJ. The effect of food on oral bioavailability of tacrolimus (FK506) in liver transplant patients[J]. Abstract Clin Pharmcol Ther,1993,5(2):229-230.
[50]钟秀英.药用p-环糊精包合物制备、检验技术研究进展[J].中药材,2003,26(4)：301-305.
[51]Hassan N, Asghar K. Investigation dchanism of β-lactam conjugated telechelic polymers of PEG and β-cyclodextrin as the new nanosized drug carrier devices[J]. Carbo Polym,2009, 76 (1) 46-50.
[52]谷福根,高永良,崔福德.环糊精包合物研究进展[J].中国新药杂志,2005,14(6)：686-693.
[53]张莉,罗来辉.环糊精包合物的研究进展[J].华北工学院学报,2003,24(4)：278-281.
[54]Thompson D O. Cyclodextrins enabling excipients:their present and future use in pharmaceuticals[J]. Crit Rev Ther Drug Carrier Syst,1997,14 (1) 101-104.
[55]Li J, Loh X J. Cyclodextrin-based supramolecular architectures:Syntheses, structures, and applications for drug and gene delivery[J]. Adv Drug Delivery Rev,2008,60 (9): 1000-1017.
[56]马开,秦文杰,高寒.痛可停胶囊挥发油p-环糊精包合物的实验研究[J].中药新药与临床药理,2003,14(6)：409-413.
[57]丁燕飞,姚瑶,周彦彬等.尼群地平p-环糊精包合物的研制[J].中国医院药学杂志,2003,23(8)：461-464.
[58]Stella VJ, He Q. Cyclodextrins[J]. Toxicol Pathol,2008,36(1):30-42.
[59]尤思路,高秀蓉,廖昌军.饱和水溶液法制备葛根素的p-环糊精包合物研究[J].西南军医,2008,10(4)：17-19.
[60]易军,李晓芳,苏建湘等.超声法制备石菖蒲挥发油p-环糊精包合物[J].时珍国医国药,2006,17(3)：381-383.
[61]齐美玲,高晶,邵清龙.大蒜素-环糊精包合物的制备及大蒜素的含量测定[J].北京理工大学学报,2004,24(7)：650-655.
[62]邵伟,袁敏,王大庆等.黄芩甙-羟丙基-p-环糊精包合物的研究[J].中成药,2001,23(5)：318-322.
[63]孙伟张,曾仁杰,景莉等.喷雾干燥法制备盐酸异丙嗪p-环糊精包合物的实验研究[J].西南国防医药,2001,1](6)385-388.
[64]Beudad H, Legrand P, Lebas G, et al. Combined hydroxyl propyleyelodextrin and poly (alkylcyanoaerylatel nanoparticles intended for oral administration of saquinavir[J]. Int J Pharm,2001,218:113-124.
[65]Francesca M, Marcos G, Paola M, et al. A new drug nanocarrier consisting of chitosan and hydoxypropylcyclodextrin[J]. Eur J Pharm Biopharm,2006,63:79-86.
[66]Alexander H K, Mana J A. Chitosan/cyclodextrin nanoparticles as macromolecular drug delivery system[J]. Int J Pharm,2007,34:134-142.
[67]尹婕,邓盛齐.环糊精及其衍生物在新型给药系统中的应用进展[J].国外医药抗生素分册,2006,27(5)：228-232.
[68]Hidetoshi A, Kiyokazu Y, Kouzou M, et al. Comparative studies of the enhancing effects of cyclodextrins on the solubility and oral bioavailability of tacrolimus in rats[J]. J Pharm Sci,2001,90 (6):690-701.
[69]陶少林.灵芝酸单体Me肠溶胶囊及药动学的研究[M].上海：华东理工大学,2009.
[70]顾磊.两种交联剂在氟尿嘧啶壳聚糖微球制备中的研究[M].上海：华东理工大学,2008.
[71]K Langer, M G Anhorn, I Steinhauser, et al. Human serum albumin (HSA) nanoparticles:Reproducibility of preparation process and kinetics of enzymatic degradation[J]. Int J Pharm,2008,347:109-117.
[72]陆彬,李凤前.白蛋白微球的热变性对其水溶性及酶降解性的影响[J].药学学报,2000,35(7)：535-538.
[73]Feng Q L, Hua S, Jing W. Preparation and characterization of sodium ferulate entrapped bovine serum albumin nanoparticles for liver targeting[J]. Int J Pharm,2008,349:274-282.
[74]王旭波,徐希明,葛亮等.半乳糖化阿霉素白蛋白纳米粒的制备及其质量评价[J].中国新药杂志,2005,14(7)：870-872.
[75]苏华,胡晋红,李凤前.表面活性剂对高水溶性药物阿魏酸钠白蛋白纳米粒的载药特性影响[J].药学服务与研究,2005,5(2)：153-156.
[76]陈悦HPLC法测定他克莫司的含量及有关物质[J].药物分析志,2008,28(2)：309-313.
[77]郑国钢,管燕,罗金文.反相高效液相色谱法测定他克莫司胶囊的含量[J].中国药业,2007,16(1)：17-18.
[78]李国琴,曾苏.梯度洗脱高效液相色谱法检测他克莫司有关物质[J].中国药学杂志,2008,43(7)：551-558.
[79]Takeshi A, Tomoko N, Masaru Y, et al. Quantitative deterruination of tautonmric FK506 by reversed-phase liquid chromatography[J]. J Pharm Biomed Anal,1996,14:339-346.
[80]梁陈方,王亚洲.反相高效液相色谱法测定他克莫司的血药浓度[J].广西医学,2009,31(3)：425-426.