摘要
目的考察乳酸-羟乙醇酸共聚物(PLGA)微球制备工艺对新城疫病毒的稳定性影响,并寻求合适的保护剂以稳定新城疫病毒的活性。方法 采用复乳-溶剂挥发法制备PLGA微球,并添加不同的保护剂进行考察。紫外分光光度法测定蛋白浓度,利用红血球凝集反应测定病毒效价。结果涡旋、高速离心和搅拌等剪切力时新城疫病毒的活性影响不大;超声时病毒活性产生一定的影响;有机溶剂对病毒活性的影响最大;加保护剂可得到一定的改善。结论制备新城疫病毒的缓释PLGA微球时,宜加冰浴超声,或缩短超声时间,延长超声间隔;选用聚乙二醇6000或羟丙-β环糊精作保护剂可显著稳定病毒活性。如果可避免有机溶剂的使用,将极大地促进新城疫病毒的活性。
Abstract
OBJECTIVE To evaluate the effects of formulation technique of PLGA microspheres on the stability of Newcastle disease virus and to look for a proper protectant to stabilize Newcastle disease virus. METHODS PLGA microspheres with or without additives were prepared by a double-emulsion solvent extraction/evaporation method. The protein concentration was determined by UV spectrophotography. The virus potency was determined by a hemagglutinin test. RESULTS The shearing forces, such as vortex, high speed centrifugation or stirring , had little influences on the activity of Newcastle disease virus. Meanwhile ultrasound had some effects, and organic solvents played the major key. The activity of virus was improved by adding some protectants. CONCLUSION To prepare the better microspheres of PLGA, ultrasound should be performed in the iced bath, and the keeping time should be shorten and the interval time should be lengthen. Polyethylene glycol 6000 or HP-β-cyclodextrin was used as a protectant to stabilize the protein. The activity of Newcastle disease virus will be raised sharply if the organic solvents are avoided.
关键词
新城疫病毒 /
微球 /
乳酸-羟乙醇酸共聚物 /
稳定性
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Key words
newcastle disease virus /
microspheres /
PLGA /
stability
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郑彩虹;许东航;梁文权.
PLGA微球制备工艺对新城疫病毒稳定性的影响[J]. 中国药学杂志, 2005, 39(22): 1724-1726
ZHENG Ci-hong;XU Dong-hng;LING Wen-qun.
Effects of formulation technique of PLGA microspheres on stability of Newcastle disease virus [J]. Chinese Pharmaceutical Journal, 2005, 39(22): 1724-1726
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参考文献
[1]刘开扬,刘芳,戴洁,等. 新城疫病毒抗癌作用及其应用的研究进展[J].中国肿瘤生物治疗杂志,2002,9(4):289.
[2]古长庆,金宁一.鸡新城疫病毒抗肿瘤作用及其机制的研究进展[J].中国肿瘤生物治疗杂志,2000,7(3):238.
[3]Phuangsab A, Lorence RM, Reichard KW, et al. Newcastle disease virus therapy of human tumor xenografts: Antitumor effects of local or systemic administration[J].Cancer Lett,2001,172(1):27.
[4]Castellanos IJ, Cruz G, Crespo R, et al. Encapsulationinduced aggregation and loss in activity of γ-chymotrypsin and their prevention[J].J Controlled Release, 2002,81:307.
[5]Zheng CH, Liang WQ, Yu HY, et al. Evaluation of different methods to determine the loading of proteins in PLGA microspheres[J]. Pharmazie,2004,59 (3):232.
[6]van de Weert, Hoechstetter J, Hennink WE, et al. The effect of a water/organic solvent interface on the structural stability of lysozyme[J].J Controlled Release,2000,68(3):351.
[7]Wang W. Instability, stabilization, and formulation of liquid protein pharmaceuticals[J].Int J Pharm,1999,185(2):129.
[8]Kang J, Schwendeman SP. Comparison of the effects of Mg(OH)2 and sucrose on the stability of bovine serum albumin encapsulated in injectable poly(d,llactidecoglycolide) implants[J].Biomaterials,2002,23(1):239.
[9]Diwan M, Park TG. Pegylation enhances protein stability stability during encapsulation in PLGA microspheres[J].J Controlled Release, 2001,73:233.
[10]Kang F, Jiang G, Hinderliter A, et al. Lysozyme stability in primary emulsion for PLGA microsphere preparation: effect of recovery methods and stabilizing excipients[J].Pharm Res, 2002,19(5):629.
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脚注
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基金
国家自然科学基金资助项目(30171113)
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