干扰素α聚乳酸乙醇酸微球的研究
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摘要
本研究以干扰素α(IFNα)为模型药物,以聚乳酸—乙醇酸(PLGA)为载体材料,采用W/O/W复乳溶剂挥发法制备了干扰素微球,并对制得微球的缓释效果及相关性质进行了评价。
在W/O/W复乳溶剂挥发法制备微球的研究中,首先以微球粒径、圆整度为指标,考察了微球制备过程中的各种影响因素,初步确定了干扰素微球的制备工艺条件。然后通过L9(3~4)正交试验设计考察了PLGA浓度、聚乙烯醇(PVA)浓度、油水相比、稳定剂种类与用量对微球质量的影响,对W/O/W法制备干扰素微球的工艺进行了优化,确定了制备干扰素微球的最佳工艺条件。
在微球缓释效果的研究中,首先对微球的体外缓释效果进行了考察。采用自行设计的体外释药方法对0.17dL/g、0.39dL/g、0.60dL/g、0.89dL/g、1.13dL/g五个不同黏度,5%、10%、15%、20%、25%五个不同浓度的PLGA制备的干扰素微球的体外缓释效果进行了比较性研究,得出了不同黏度、不同浓度的PLGA微球缓释时间的长短,进一步完善了微球的最佳制备工艺。结果表明15%的0.89dL/g PLGA干扰素微球的缓释效果较好,其体外释放能很好的符合缓释制剂的特征,Higuchi方程为Q=2.2034t~(1.2)+25.605,r=0.9023,15天的累积释放量约为75%。对此微球的体外释放液中的干扰素的生物学活性进行考查,发现其一周的体外释放液都有很好的生物学活性。以大鼠为研究对象,对干扰素微球的体内缓释效果进行考察,干扰素微球具有明显的缓释作用,生物利用度比干扰素粉针提高了2倍。
本研究还对15%的0.89dL/g PLGA微球的理化性质与生物学性质进行评价。结果干扰素微球的球形圆整,不黏连,粒径分布较均匀,平均粒径为65.84μm。干扰素微球的堆密度为0.1856g/ml;休止角为29.42°,流动性较好:吸湿性非常小,在90%相对湿度的环境中,吸湿增重比不大于1%,但是湿度会使干扰素的含量下降,所以干扰素微球应该密封干燥保存。DSC分析结果表明干扰素被包裹在PLGA微球中,而不是黏附在微球表面。顶空—气质联用色谱法测定了微球中残留的二氯甲烷量,得出微球有机溶剂残留量符合药典规定。微球的生物学性质研究主要是指生物降解性和生物相容性研究。在微球的生物降解性研究中,对其体内、外降解情况都进行了研究,结果微球在体内降解速率比在体外降解速率要快,在体内6周可以完全降解,不会在体内造成蓄积。对微球的体内组织相容性进行研究,大鼠肌肉注射一定的微球,在6周内观察,注射部位肌肉无充血,无水肿,触之无包块,对机体无明显刺激性反应,说明微球具有良好的生物相容性。本研究为多肽、蛋白质类药物新制剂的开发开辟了道路,为微球制剂的研究提供了理论技术指导。根据目前的研究结果,如再予以更深入细致的研究,干扰素微球剂有望成为临床治疗乙肝、丙肝等疾病的一种新剂型。
Interferon-alpha(IFNα)was chosen as a model drug and poly(lactic acid-glycolic acid) (PLGA)was used as the carrier to prepare Interferon-alpha sustained-release microspheres by W/O/W double emulsions solvent evaporation method.Furthermore,the drug release behaviors and the properties of IFNαmicrospheres were deeply studied.
During the process of IFNαmicrospheres preparation,the effect of various factors were studied at first with the particle's size and roudness of the microspheres as the criteria.Then the optimum formulation of IFNαmicrospheres prepared by W/O/W double emulsions solvent evaporation method was determined by employing L9(3~4)orthogonal test design,including the effect of PLGA concentration,polyvinyl alcohol(PVA) concentration,oil/water ratio,and choice of stabilizers and quantity used.
In the sustained-release study of IFNαmicrospheres,the in vitro release behaviors of IFNαmicrospheres were studied firstly by an improved release method.We compared the in vitro release profiles of IFNαmicrospheres,which prepared by five different viscosities, 0.17dL/g、0.39dL/g、0.60dL/g、0.89dL/g、1.13dL/g and five different concentrations,5%、10%、15%、20%、25%.The optimum formulation of IFNαmicrospheres preparation was also further perfected by the results of the in vitro release.In vitro release profile of IFNαmicrospheres prepared by 15%0.89dL/g PLGA met the criteria of sustained release formulation preferably and it was able to prolong drug action.Higuchi equation was Q=2.203t~(1/2)+25.605,r=0.9023.The accumulated amount of IFNαreleased was 75%in 15 days.Moreover,the biological activity of IFNαwas measured by Cytopathic Inhibition Assay suggesting that bioactive IFNαhad been firmly released from microsphere for more than 7 days.The pharmacokinetic experiments of this microspheres showed an obvious sustained fashion,and the relative bioavailability of IFNαmicrospheres to IFNαwas about 200%.
The IFNαmicrospheres prepared by optimum formulation were white powder with fine fluidity.The surface morphology of microspheres had a spherical shape with a smooth and porous surface.Particle size of IFNαmicrospheres were nearly normality and the average diameter of microspheres was 65.84μm.The bulk density and the angel of repose of microspheres were 0.1856g/ml and 29.42°respectively.The moisture absorption of microspheres was low for their hygroscopic ratio less than 1%under the condition of 90% relative humidity.However,IFNαmicrospheres should be stored in airproof and dry container since humidity could accelerate IFNαdegradation.The DSC curve of microspheres confirmed that IFNαwas loaded within the microspheres,rather than adsorbed on their surfaces.For dichloromethane was used to prepare microspheres,the residual quantity of dichloromethane in microspheres should be measured.The amount determined by gas chromatography and mass spectra was lower than the criteria of Chinese Pharmacopeia.The biodegradability of PLGA microspheres was investigated by an improved method in vitro and by intramuscular injection in rats in vivo.The biodegradable speeds of IFNαmicrospheres in vivo were faster than those in vitro.After six weeks,the degradable percentage of IFNαmicrospheres was 80%in vitro and 100%in vivo,so that IFNαmicrospheres would not be accumulated in body.Biocompatibility of microspheres had been observed for six weeks by intramuscular injection in rats.It showed that IFNαmicrospheres were biocompatible for only slight inflammatory response but no pathological change at the site of injection.
The results of the above study provided a useful exploration for the experimental methods and theories of peptide and protein microspheres and a scientific basis for developing of IFNαmicrospheres as a new drug to cure hepatitis.
在W/O/W复乳溶剂挥发法制备微球的研究中,首先以微球粒径、圆整度为指标,考察了微球制备过程中的各种影响因素,初步确定了干扰素微球的制备工艺条件。然后通过L9(3~4)正交试验设计考察了PLGA浓度、聚乙烯醇(PVA)浓度、油水相比、稳定剂种类与用量对微球质量的影响,对W/O/W法制备干扰素微球的工艺进行了优化,确定了制备干扰素微球的最佳工艺条件。
在微球缓释效果的研究中,首先对微球的体外缓释效果进行了考察。采用自行设计的体外释药方法对0.17dL/g、0.39dL/g、0.60dL/g、0.89dL/g、1.13dL/g五个不同黏度,5%、10%、15%、20%、25%五个不同浓度的PLGA制备的干扰素微球的体外缓释效果进行了比较性研究,得出了不同黏度、不同浓度的PLGA微球缓释时间的长短,进一步完善了微球的最佳制备工艺。结果表明15%的0.89dL/g PLGA干扰素微球的缓释效果较好,其体外释放能很好的符合缓释制剂的特征,Higuchi方程为Q=2.2034t~(1.2)+25.605,r=0.9023,15天的累积释放量约为75%。对此微球的体外释放液中的干扰素的生物学活性进行考查,发现其一周的体外释放液都有很好的生物学活性。以大鼠为研究对象,对干扰素微球的体内缓释效果进行考察,干扰素微球具有明显的缓释作用,生物利用度比干扰素粉针提高了2倍。
本研究还对15%的0.89dL/g PLGA微球的理化性质与生物学性质进行评价。结果干扰素微球的球形圆整,不黏连,粒径分布较均匀,平均粒径为65.84μm。干扰素微球的堆密度为0.1856g/ml;休止角为29.42°,流动性较好:吸湿性非常小,在90%相对湿度的环境中,吸湿增重比不大于1%,但是湿度会使干扰素的含量下降,所以干扰素微球应该密封干燥保存。DSC分析结果表明干扰素被包裹在PLGA微球中,而不是黏附在微球表面。顶空—气质联用色谱法测定了微球中残留的二氯甲烷量,得出微球有机溶剂残留量符合药典规定。微球的生物学性质研究主要是指生物降解性和生物相容性研究。在微球的生物降解性研究中,对其体内、外降解情况都进行了研究,结果微球在体内降解速率比在体外降解速率要快,在体内6周可以完全降解,不会在体内造成蓄积。对微球的体内组织相容性进行研究,大鼠肌肉注射一定的微球,在6周内观察,注射部位肌肉无充血,无水肿,触之无包块,对机体无明显刺激性反应,说明微球具有良好的生物相容性。本研究为多肽、蛋白质类药物新制剂的开发开辟了道路,为微球制剂的研究提供了理论技术指导。根据目前的研究结果,如再予以更深入细致的研究,干扰素微球剂有望成为临床治疗乙肝、丙肝等疾病的一种新剂型。
Interferon-alpha(IFNα)was chosen as a model drug and poly(lactic acid-glycolic acid) (PLGA)was used as the carrier to prepare Interferon-alpha sustained-release microspheres by W/O/W double emulsions solvent evaporation method.Furthermore,the drug release behaviors and the properties of IFNαmicrospheres were deeply studied.
During the process of IFNαmicrospheres preparation,the effect of various factors were studied at first with the particle's size and roudness of the microspheres as the criteria.Then the optimum formulation of IFNαmicrospheres prepared by W/O/W double emulsions solvent evaporation method was determined by employing L9(3~4)orthogonal test design,including the effect of PLGA concentration,polyvinyl alcohol(PVA) concentration,oil/water ratio,and choice of stabilizers and quantity used.
In the sustained-release study of IFNαmicrospheres,the in vitro release behaviors of IFNαmicrospheres were studied firstly by an improved release method.We compared the in vitro release profiles of IFNαmicrospheres,which prepared by five different viscosities, 0.17dL/g、0.39dL/g、0.60dL/g、0.89dL/g、1.13dL/g and five different concentrations,5%、10%、15%、20%、25%.The optimum formulation of IFNαmicrospheres preparation was also further perfected by the results of the in vitro release.In vitro release profile of IFNαmicrospheres prepared by 15%0.89dL/g PLGA met the criteria of sustained release formulation preferably and it was able to prolong drug action.Higuchi equation was Q=2.203t~(1/2)+25.605,r=0.9023.The accumulated amount of IFNαreleased was 75%in 15 days.Moreover,the biological activity of IFNαwas measured by Cytopathic Inhibition Assay suggesting that bioactive IFNαhad been firmly released from microsphere for more than 7 days.The pharmacokinetic experiments of this microspheres showed an obvious sustained fashion,and the relative bioavailability of IFNαmicrospheres to IFNαwas about 200%.
The IFNαmicrospheres prepared by optimum formulation were white powder with fine fluidity.The surface morphology of microspheres had a spherical shape with a smooth and porous surface.Particle size of IFNαmicrospheres were nearly normality and the average diameter of microspheres was 65.84μm.The bulk density and the angel of repose of microspheres were 0.1856g/ml and 29.42°respectively.The moisture absorption of microspheres was low for their hygroscopic ratio less than 1%under the condition of 90% relative humidity.However,IFNαmicrospheres should be stored in airproof and dry container since humidity could accelerate IFNαdegradation.The DSC curve of microspheres confirmed that IFNαwas loaded within the microspheres,rather than adsorbed on their surfaces.For dichloromethane was used to prepare microspheres,the residual quantity of dichloromethane in microspheres should be measured.The amount determined by gas chromatography and mass spectra was lower than the criteria of Chinese Pharmacopeia.The biodegradability of PLGA microspheres was investigated by an improved method in vitro and by intramuscular injection in rats in vivo.The biodegradable speeds of IFNαmicrospheres in vivo were faster than those in vitro.After six weeks,the degradable percentage of IFNαmicrospheres was 80%in vitro and 100%in vivo,so that IFNαmicrospheres would not be accumulated in body.Biocompatibility of microspheres had been observed for six weeks by intramuscular injection in rats.It showed that IFNαmicrospheres were biocompatible for only slight inflammatory response but no pathological change at the site of injection.
The results of the above study provided a useful exploration for the experimental methods and theories of peptide and protein microspheres and a scientific basis for developing of IFNαmicrospheres as a new drug to cure hepatitis.
引文
1. Sinha VR, Trehan A: Biodegradable microspheres for protein delivery. J Control Release 2003, 90(3):261-280.
2. Wang ZY, Zhao YM, Wang F, Wang J: Syntheses of poly(lactic acid-co-glycolic acid) Serial Biodegradable Polymer Materials via Direct Melt Polycondensation and Their Characterization. J Applied Polymer Science 2006, 99(2):244-252.
3. Mahmutovic S, Beslagic E: Signifinance of the interferon(IFN) in the therapy. Bosn J Basic Med Sci 2004, 4(4):42-44.
4. Isaacs A, Lindenmann J: Virus interference. I. The interferon. Proc R Soc Lond B Biol Sci 1957, 147(927):258-267.
5. De Andrea M, Ravera R, Gioia D, Gariglio M, Landolfo S: The interferon system:an overview. Eur J Paediatr Neurol 2002, 6 Suppl A:A41 -46.
6. Cleland JL, Jones LS: Stable formulations of recombinant human growth hormone and interferon-γ for microencapsulation in biodegradable microspheres. Pharm Res 1996, 13(10):1464-1468.
7. Yang J, Cleland JL: Factors affecting the in vitro release of recombinant human interferon-gamma (rhIFN-gamma) from PLGA microspheres .J Pharm Sci 1997, 86(8):908-914.
8. Kajihara M, Sugie T, Mizuno M, Tamura N, Sano A, Fujioka K, Kashiwazaki Y, Yamaoka T, Sugawara S, Urabe Y: Development of new drug delivery system for protein drugs using silicone (I). J Control Release 2000, 66(1 ):49-61.
9. Yamagata Y, Iga K, Ogawa Y: Novel sustained-release dosage forms of proteins using polyglycerol esters of fatty acids. J Control Release 2000, 63(3):319-329.
10. Van Slooten ML, Boerman O, Romoren K, Kedar E, Crommelin DJ, Storm G: Liposomes as sustained release system for human interferon-gamma: biopharmaceutical aspects. Biochim Biophys Acta 2001, 1530(2-3): 134-145.
11. Bailon P, Palleroni A, Schaffer CA, Spence CL, Fung WJ, Porter JE, Ehrlich GK, Pan W, Xu ZX, Modi MW et al: Rational design of a potent, long-lasting form of interferon: a 40 kDa branched polyethylene glycol-conjugated interferon alpha-2a for the treatment of hepatitis C. Bioconjug Chem 2001,12(2):195-202.
12. Guiziou B, Armstrong DJ, Elliott PN, Ford JL, Rostron C: Investigation of in-vitro release characteristics of NSAID-loaded polylactic acid microspheres. J Microencapsul 1996, 13(6):701-708.
13. O'Donnell PB, McGinity JW: Influence of processing on the stability and release properties of biodegradable microspheres containing thioridazine hydrochloride. Eur J Pharm Biopharm 1998, 45(1):83-94.
14. Jeyanthi R, Mehta RC, Thanoo BC, DeLuca PP: Effect of processing parameters on the properties of peptide-containing PLGA microspheres. J Microencapsul 1997,14(2): 163-174.
15. Uchida T, Goto S: Oral delivery of poly(lactide-co-glycolide) microspheres containing ovalbumin as vaccine formulation: particle size study. Biol Pharm Bull 1994,17(9): 1272-1276.
16. Cleland JL: Solvent evaporation processes for the production of controlled release biodegradable microsphere formulations for therapeutics and vaccines. Biotechnol Prog 1998, 14(1):102-107.
17. Chang AC, Gupta RK: Stabilization of tetanus toxoid in poly(DL-lactic-co-glycolic acid) microspheres for the controlled release of antigen. J Pharm Sci 1996, 85(2):129-132.
18. Sampath SS, Garvin K, Robinson DH: Preparation and characterization of biodegradable poly(L-lactic acid) gentamicin delivery systems. Int J Pharm 1992, 78(2-3): 165-174.
19. Conti B, Genta 1, Modena T, Pavanetto F: Investigation on process parameters involved in polylactide-co-glycolide microspheres preparation.. Drug Dev Ind Pharm 1995, 21(5):615-622.
20. Carrascosa C, Torres-Aleman I, Lopez-Lopez C, Carro E, Espejo L, Torrado S, Torrado JJ: Microspheres containing insulin-like growth factor I for treatment of chronic neurodegeneration. Biomaterials 2004, 25(4):707-714.
21. Wang N, Wu XS: A novel approach to stabilization of protein drugs in poly(lactide-co-glycolic acid) microspheres using agarose hydrogel. . Int J Pharm 1998, 166( I): 1-14.
22. Thomasin C, Corradin G, Men Y, Merkle HP, Gander B: Tetanus toxoid and synthetic malaria antigen containing poly(lactide)/ poly(lactide-co-glycolide) microspheres : importance of polymer degradation and antigen immune response.. J Control Release 1996, 41(1-2): 131 -145.
23. Bleich J, Mueller BW, Wassmus W: Aerosol solvent extraction system. A new microparticle production technique. Int J Pharm 1993, 97( 1 -3): 111 -117.
24. Hayashi Y, Yoshioka S, Aso Y, Li Wan Po A, Terao T: Entrapment of proteins in poly(L-lactide) microspheres using reversed micelle solvent evaporation. Pharm Res 1994, 11(2):337-340.
25. Ruiz JM, Tissier B, Benoit JP: Microencapsulation of peptide: a study of the phase separathion of poly(D,L-lacticacid-co-glycolic acid) copolymers 50/50 by Silicone oil. . Int J Pharm 1989, 49(1):69-77.
26. McGee JP, Davis SS, O'Hagan DT: Zero order release of protein from poly(D,L-lactide-co-glycolide) microparticles prepared using a modified phase separation technique. . J Control Release 1995, 34(2):77-86.
27. Castellanos IJ, Cuadrado WO, Griebenow K: Prevention of structural perturbations and aggregation upon encapsulation of bovine serum albumin into poly(lactide-co-glycolide) micropheres using the solid-in-oil-in water technique. J Pharm Pharmacol 2001, 53(8): 1099-1107.
28. Park TG, Yong Lee H, Sung Nam Y: A new preparation method for protein loaded poly(D, L-lactic-co-glycolic acid) microspheres and protein release mechanism study. J Control Release 1998, 55(2-3): 181-191.
29. Mehta RC, Thanoo BC, DeLuca PP: Peptide containing microspheres from low molecular weight andhydrophilic poly(d,l-lactide-co-glycolide). J Control Release 1996, 41(3):249-257.
30. Crotts G, Sah H, Park TG: Absorption determines in vitro protein release rate from biodegradable microspheres : quantitative analysis of analysis surface area during degradation. J Control Release 1997, 47( 1): 101 -111.
31. Uchida T, Yoshida K, Ninomiya A, Goto S: Optimization of preparative conditions for polylactide (PLA) microspheres containing ovalbumin. Chem Pharm Bull (Tokyo) 1995, 43(9): 1569-1573.
32. Rojas J, Pinto-Alphandary H, Leo E, Pecquet S, Couvreur P, Fattal E: Optimization of the encapsulation and release of beta-lactoglobulin entrapped poly(DL-lactide-co-glycolide) microspheres. Int J Pharm 1999, 183(1):67-71.
33. Rosa GD, Iommelli R, La Rotonda MI, Miro A, Quaglia F: Influence of the co-encapsulation of different non-ionic surfactants on the properties of PLGA insulin-loaded microspheres. J Control Release 2000, 69(2):283-295.
34. Joyanthi R, Thanoo BC, Metha RC, DeLuca PP: Effect of solvent removal technique on the metrix characteristics of polylactide/glycolide microspheres for peptide delivery. . J Control Release 1996, 38(2):235-244.
35. Yang YY, Chia HH, Chung TS: Effect of preparation temperature on the characteristics and release profiles of PLGA microspheres containing protein fabricated by double-emulsion solvent extraction/evaporation method. J Control Release 2000, 69(1):81-96.
36. Wasington C: Drug release from microspheres systems: a critical review. . Int J Pharm 1990, 58:1-8.
37. Sanchez A, Tobio M, Gonzalez L, Fabra A, Alonso MJ: Biodegradable micro- and nanoparticles as long-term delivery vehicles for interferon-alpha. Eur J Pharm Sci 2003, 18(3-4):221-229.
38. Kim HK, Park TG: Comparative study on sustained release of human growth hormone from semi-crystalline poly(L-lactic acid) and amorphous poly(D,L-lactic-co-glycolic acid) microspheres:morphological effect on protein release,J Control Release 2004,98(1):115-125.
39.Park TG,Lu WQ,Crotts G:Important of in vitro experimental conditions on protein release kinetics,stability and polymer degradation in protein encapsulated poly(D,L-lactic-co-glycolic acid)microspheres.,J Control Release 1995,33(2):211-222.
40.Chert L,Apte RN,Cohen S:Characterization of PLGA microspheres for the controlled delivery of IL-1 a for tumor immunotherapy,J Control Release 1997,43(2):261-272.
41.国家药典委员会编.:中华人民共和国药典,第三部edn:化学工业出版社;2005.:附录XC.
42.Li X,Deng X,Yuan M,Xiong C,Huang Z,Zhang Y,Jia W:Investigation on process parameters involved in preparation of poly-DL-lactide-poly(ethylene glycol)microspheres containing Leptospira Interrogans antigens.Int J Pharm 1999,178(2):245-255.
43.Kim TH,Park TG:Critical effect of freezing/freeze-drying on sustained release of F1TC-dextran encapsulated within PLGA microspheres.Int J Pharm 2004,271(1-2):207-214.
44.Pistner H,Bendix DR,Muhling J,Reuther JF:Poly(L-lactide):a long-term degradation study in vivo.Part Ⅲ.Analytical characterization.Biomaterials 1993,14(4):291-298.
45.Cohen S,Yoshioka T,Lucarelli M,Hwang LH,Langer R:Controlled delivery systems for proteins based on poly(lactic/glycolic acid)microspheres.Pharm Res 1991,8(6):713-720.
46.Okada H,Toguchi H:Biodegradable microspheres in drug delivery.Crit Rev Ther Drug Carrier Syst 1995,12(1):1-99.
47.Baras B,Benoit MA,Gillard J:Parameters influencing the antigen release from spray-dried poly(DL-lactide)microparticles.Int J Pharm 2000,200(1):133-145.
48.Ogawa Y,Yamamoto M,Takada S,Okada H,Shimamoto T:Controlled-release of leuprolide acetate from polylactic acid or copoly(lactic/glycolic)acid microcapsules:influence of molecular weight and copolymer ratio of polymer.Chem Pharm Bull(Tokyo)1988,36(4):1502-1507.
49.Shah VP,Tsong Y,Sathe P,Liu JP:In vitro dissolution profile comparison-statistics and analysis of the similarity factor,f2.Pharm Res 1998,15(6):889-896.
50.Gallagher KM,Corrigan OI:Mechanistic aspects of the release of levamisole hydrochloride from biodegradable polymers.J Control Release 2000,69(2):261-272.
51.Shenderova A,Burke TG,Schwendeman SP:Stabilization of 10-hydroxycamptothecin in poly(lactide-co-glycolide)microsphere delivery vehicles.Pharm Res 1997,14(10):1406-1414.
52.Ravivarapu HB,Burton K,DeLuca PP:Polymer and microsphere blending to alter the release of a peptide from PLGA microspheres.Eur J Pharm Biopharm 2000,50(2):263-270.
53.Heller J:Controlled drug release from moomlithic bioerodeble Polymer devicesl..Pharmacy lnt 1986,11:310-318.
54.Endoh H,Kawaguchi T,Seki T,Hasegawa T,Juni K:Controlled release of 5-fluoro-2'-deoxyuridine by the combination of prodrug and polymer matrix.Chem Pharm Bull (Tokyo)1991,39(2):458-464.
55.Kaniwa N,Shameen M,Katori N:The suitable temperature for the evaluation of controlled release products in vitro.Pharmazie 1995,50(2):53-55.
56.Soppimath KS,Aminabhavi TM,Kulkarni AR,Rudzinski WE:Biodegradable polymeric nanoparticles as drug delivery devices.J Control Release 2001,70(1-2):1-20.
57.Oosha E Muller Rh,Davis SS:A continous flow system for in vitro evaluation of drug-loaded biodegradable colloidal carriers.J Pharm Sci 1988,40(6):138-145.
58.杨帆.,张永明.,赵耀明.,李丹涛.,宋风兰.,徐安龙.:干扰素微球的制备及其体外释药性能研究.中国医院药学杂志2005,25(9):807-809.
59.Vert M,Mauduit J,Li S:Biodegradation of PLA/GA polymers:increasing complexity.Biomaterials 1994,15(15):1209-1213.
60.Schakenraad JM,Hardonk MJ,Feijen J,Molenaar 1,Nieuwenhuis P:Enzymatic activity toward poly(L-lactic acid)implants.J Biomed Mater Res 1990,24(5):529-545.
61.lsobe M,Yamazaki Y,Oida S:Bone morphogenetic protein encapsulation with a biodegradable and biocompatible polymer.J Biomed Mater Res 1996,32(10):433-438.
62.Kamei S,Inoue Y:Phagocytosis of microspheres by macrophages.Adv Polym Sci 1990,94(5):107-141.
63.Visscher GE,Pearson JE,Fong JW,Argentieri G J,Robison RL,Maulding HV:Effect of particle size on the in vitro and in vivo degradation rates of poly(DL-lactide-co-glycolide)microcapsules.J Biomed Mater Res 1988,22(8):733-746.
64.Park TG:Degradation of poly(lactic-co-glycolic acid)microspheres:effect of copolymer composition.Biomaterials 1995,16(15):1123-1130.
65.Cadee JA,Brouwer LA,den Otter W,Hennink WE,van Luyn MJ:A comparative biocompatibility study of microspheres based on crosslinked dextran or poly(lactic-co-glycolic)acid after subcutaneous injection in rats.J Biomed Mater Res 2001,56(4):600-609.
66.Shive MS,Anderson JM:Biodegradation and biocompatibility of PLA and PLGA microspheres.Adv Drug Deliv Rev 1997,28(1):5-24.
67.Visscher GE,Robison RL,Maulding HV,Fong JW,Pearson JE,Argentieri G J:Biodegradation of and tissue reaction to 50:50 poly(DL-lactide-co-glycolide)microcapsules.J Biomed Mater Res 1985,19(3):349-365.
68.Visscher GE,Robison RL,Maulding HV,Fong JW,Pearson JE,Argentieri G J:Biodegradation of and tissue reaction to poly(DL-lactide)microcapsules.J Biomed Mater Res 1986,20(5):667-676.
69. Visscher GE, Robison MA, Argentieri GJ: Tissue response to biodegradable injectable microcapsules. J Biomater Appl 1987, 2( 1): 118-131.
70. Yanaguchi K, Anderson JM: Biocompatibility studies of naltrexone sustained release formulations. J Control Release 1992, 19(2):299-314.
71. Grizzi I, Garreau H, Li S, Vert M: Hydrolytic degradation of devices based on poly(DL-lactic acid) size-dependence. Biomaterials 1995, 16(4):305-311.
72. Bostman O: Intense granulomatous in inflammatory lesions associated with absorbable internal fixation devices made of polyglycolide in ankle fractures. Clin Orthop Relat Res 1992, 778(5): 178-199.
2. Wang ZY, Zhao YM, Wang F, Wang J: Syntheses of poly(lactic acid-co-glycolic acid) Serial Biodegradable Polymer Materials via Direct Melt Polycondensation and Their Characterization. J Applied Polymer Science 2006, 99(2):244-252.
3. Mahmutovic S, Beslagic E: Signifinance of the interferon(IFN) in the therapy. Bosn J Basic Med Sci 2004, 4(4):42-44.
4. Isaacs A, Lindenmann J: Virus interference. I. The interferon. Proc R Soc Lond B Biol Sci 1957, 147(927):258-267.
5. De Andrea M, Ravera R, Gioia D, Gariglio M, Landolfo S: The interferon system:an overview. Eur J Paediatr Neurol 2002, 6 Suppl A:A41 -46.
6. Cleland JL, Jones LS: Stable formulations of recombinant human growth hormone and interferon-γ for microencapsulation in biodegradable microspheres. Pharm Res 1996, 13(10):1464-1468.
7. Yang J, Cleland JL: Factors affecting the in vitro release of recombinant human interferon-gamma (rhIFN-gamma) from PLGA microspheres .J Pharm Sci 1997, 86(8):908-914.
8. Kajihara M, Sugie T, Mizuno M, Tamura N, Sano A, Fujioka K, Kashiwazaki Y, Yamaoka T, Sugawara S, Urabe Y: Development of new drug delivery system for protein drugs using silicone (I). J Control Release 2000, 66(1 ):49-61.
9. Yamagata Y, Iga K, Ogawa Y: Novel sustained-release dosage forms of proteins using polyglycerol esters of fatty acids. J Control Release 2000, 63(3):319-329.
10. Van Slooten ML, Boerman O, Romoren K, Kedar E, Crommelin DJ, Storm G: Liposomes as sustained release system for human interferon-gamma: biopharmaceutical aspects. Biochim Biophys Acta 2001, 1530(2-3): 134-145.
11. Bailon P, Palleroni A, Schaffer CA, Spence CL, Fung WJ, Porter JE, Ehrlich GK, Pan W, Xu ZX, Modi MW et al: Rational design of a potent, long-lasting form of interferon: a 40 kDa branched polyethylene glycol-conjugated interferon alpha-2a for the treatment of hepatitis C. Bioconjug Chem 2001,12(2):195-202.
12. Guiziou B, Armstrong DJ, Elliott PN, Ford JL, Rostron C: Investigation of in-vitro release characteristics of NSAID-loaded polylactic acid microspheres. J Microencapsul 1996, 13(6):701-708.
13. O'Donnell PB, McGinity JW: Influence of processing on the stability and release properties of biodegradable microspheres containing thioridazine hydrochloride. Eur J Pharm Biopharm 1998, 45(1):83-94.
14. Jeyanthi R, Mehta RC, Thanoo BC, DeLuca PP: Effect of processing parameters on the properties of peptide-containing PLGA microspheres. J Microencapsul 1997,14(2): 163-174.
15. Uchida T, Goto S: Oral delivery of poly(lactide-co-glycolide) microspheres containing ovalbumin as vaccine formulation: particle size study. Biol Pharm Bull 1994,17(9): 1272-1276.
16. Cleland JL: Solvent evaporation processes for the production of controlled release biodegradable microsphere formulations for therapeutics and vaccines. Biotechnol Prog 1998, 14(1):102-107.
17. Chang AC, Gupta RK: Stabilization of tetanus toxoid in poly(DL-lactic-co-glycolic acid) microspheres for the controlled release of antigen. J Pharm Sci 1996, 85(2):129-132.
18. Sampath SS, Garvin K, Robinson DH: Preparation and characterization of biodegradable poly(L-lactic acid) gentamicin delivery systems. Int J Pharm 1992, 78(2-3): 165-174.
19. Conti B, Genta 1, Modena T, Pavanetto F: Investigation on process parameters involved in polylactide-co-glycolide microspheres preparation.. Drug Dev Ind Pharm 1995, 21(5):615-622.
20. Carrascosa C, Torres-Aleman I, Lopez-Lopez C, Carro E, Espejo L, Torrado S, Torrado JJ: Microspheres containing insulin-like growth factor I for treatment of chronic neurodegeneration. Biomaterials 2004, 25(4):707-714.
21. Wang N, Wu XS: A novel approach to stabilization of protein drugs in poly(lactide-co-glycolic acid) microspheres using agarose hydrogel. . Int J Pharm 1998, 166( I): 1-14.
22. Thomasin C, Corradin G, Men Y, Merkle HP, Gander B: Tetanus toxoid and synthetic malaria antigen containing poly(lactide)/ poly(lactide-co-glycolide) microspheres : importance of polymer degradation and antigen immune response.. J Control Release 1996, 41(1-2): 131 -145.
23. Bleich J, Mueller BW, Wassmus W: Aerosol solvent extraction system. A new microparticle production technique. Int J Pharm 1993, 97( 1 -3): 111 -117.
24. Hayashi Y, Yoshioka S, Aso Y, Li Wan Po A, Terao T: Entrapment of proteins in poly(L-lactide) microspheres using reversed micelle solvent evaporation. Pharm Res 1994, 11(2):337-340.
25. Ruiz JM, Tissier B, Benoit JP: Microencapsulation of peptide: a study of the phase separathion of poly(D,L-lacticacid-co-glycolic acid) copolymers 50/50 by Silicone oil. . Int J Pharm 1989, 49(1):69-77.
26. McGee JP, Davis SS, O'Hagan DT: Zero order release of protein from poly(D,L-lactide-co-glycolide) microparticles prepared using a modified phase separation technique. . J Control Release 1995, 34(2):77-86.
27. Castellanos IJ, Cuadrado WO, Griebenow K: Prevention of structural perturbations and aggregation upon encapsulation of bovine serum albumin into poly(lactide-co-glycolide) micropheres using the solid-in-oil-in water technique. J Pharm Pharmacol 2001, 53(8): 1099-1107.
28. Park TG, Yong Lee H, Sung Nam Y: A new preparation method for protein loaded poly(D, L-lactic-co-glycolic acid) microspheres and protein release mechanism study. J Control Release 1998, 55(2-3): 181-191.
29. Mehta RC, Thanoo BC, DeLuca PP: Peptide containing microspheres from low molecular weight andhydrophilic poly(d,l-lactide-co-glycolide). J Control Release 1996, 41(3):249-257.
30. Crotts G, Sah H, Park TG: Absorption determines in vitro protein release rate from biodegradable microspheres : quantitative analysis of analysis surface area during degradation. J Control Release 1997, 47( 1): 101 -111.
31. Uchida T, Yoshida K, Ninomiya A, Goto S: Optimization of preparative conditions for polylactide (PLA) microspheres containing ovalbumin. Chem Pharm Bull (Tokyo) 1995, 43(9): 1569-1573.
32. Rojas J, Pinto-Alphandary H, Leo E, Pecquet S, Couvreur P, Fattal E: Optimization of the encapsulation and release of beta-lactoglobulin entrapped poly(DL-lactide-co-glycolide) microspheres. Int J Pharm 1999, 183(1):67-71.
33. Rosa GD, Iommelli R, La Rotonda MI, Miro A, Quaglia F: Influence of the co-encapsulation of different non-ionic surfactants on the properties of PLGA insulin-loaded microspheres. J Control Release 2000, 69(2):283-295.
34. Joyanthi R, Thanoo BC, Metha RC, DeLuca PP: Effect of solvent removal technique on the metrix characteristics of polylactide/glycolide microspheres for peptide delivery. . J Control Release 1996, 38(2):235-244.
35. Yang YY, Chia HH, Chung TS: Effect of preparation temperature on the characteristics and release profiles of PLGA microspheres containing protein fabricated by double-emulsion solvent extraction/evaporation method. J Control Release 2000, 69(1):81-96.
36. Wasington C: Drug release from microspheres systems: a critical review. . Int J Pharm 1990, 58:1-8.
37. Sanchez A, Tobio M, Gonzalez L, Fabra A, Alonso MJ: Biodegradable micro- and nanoparticles as long-term delivery vehicles for interferon-alpha. Eur J Pharm Sci 2003, 18(3-4):221-229.
38. Kim HK, Park TG: Comparative study on sustained release of human growth hormone from semi-crystalline poly(L-lactic acid) and amorphous poly(D,L-lactic-co-glycolic acid) microspheres:morphological effect on protein release,J Control Release 2004,98(1):115-125.
39.Park TG,Lu WQ,Crotts G:Important of in vitro experimental conditions on protein release kinetics,stability and polymer degradation in protein encapsulated poly(D,L-lactic-co-glycolic acid)microspheres.,J Control Release 1995,33(2):211-222.
40.Chert L,Apte RN,Cohen S:Characterization of PLGA microspheres for the controlled delivery of IL-1 a for tumor immunotherapy,J Control Release 1997,43(2):261-272.
41.国家药典委员会编.:中华人民共和国药典,第三部edn:化学工业出版社;2005.:附录XC.
42.Li X,Deng X,Yuan M,Xiong C,Huang Z,Zhang Y,Jia W:Investigation on process parameters involved in preparation of poly-DL-lactide-poly(ethylene glycol)microspheres containing Leptospira Interrogans antigens.Int J Pharm 1999,178(2):245-255.
43.Kim TH,Park TG:Critical effect of freezing/freeze-drying on sustained release of F1TC-dextran encapsulated within PLGA microspheres.Int J Pharm 2004,271(1-2):207-214.
44.Pistner H,Bendix DR,Muhling J,Reuther JF:Poly(L-lactide):a long-term degradation study in vivo.Part Ⅲ.Analytical characterization.Biomaterials 1993,14(4):291-298.
45.Cohen S,Yoshioka T,Lucarelli M,Hwang LH,Langer R:Controlled delivery systems for proteins based on poly(lactic/glycolic acid)microspheres.Pharm Res 1991,8(6):713-720.
46.Okada H,Toguchi H:Biodegradable microspheres in drug delivery.Crit Rev Ther Drug Carrier Syst 1995,12(1):1-99.
47.Baras B,Benoit MA,Gillard J:Parameters influencing the antigen release from spray-dried poly(DL-lactide)microparticles.Int J Pharm 2000,200(1):133-145.
48.Ogawa Y,Yamamoto M,Takada S,Okada H,Shimamoto T:Controlled-release of leuprolide acetate from polylactic acid or copoly(lactic/glycolic)acid microcapsules:influence of molecular weight and copolymer ratio of polymer.Chem Pharm Bull(Tokyo)1988,36(4):1502-1507.
49.Shah VP,Tsong Y,Sathe P,Liu JP:In vitro dissolution profile comparison-statistics and analysis of the similarity factor,f2.Pharm Res 1998,15(6):889-896.
50.Gallagher KM,Corrigan OI:Mechanistic aspects of the release of levamisole hydrochloride from biodegradable polymers.J Control Release 2000,69(2):261-272.
51.Shenderova A,Burke TG,Schwendeman SP:Stabilization of 10-hydroxycamptothecin in poly(lactide-co-glycolide)microsphere delivery vehicles.Pharm Res 1997,14(10):1406-1414.
52.Ravivarapu HB,Burton K,DeLuca PP:Polymer and microsphere blending to alter the release of a peptide from PLGA microspheres.Eur J Pharm Biopharm 2000,50(2):263-270.
53.Heller J:Controlled drug release from moomlithic bioerodeble Polymer devicesl..Pharmacy lnt 1986,11:310-318.
54.Endoh H,Kawaguchi T,Seki T,Hasegawa T,Juni K:Controlled release of 5-fluoro-2'-deoxyuridine by the combination of prodrug and polymer matrix.Chem Pharm Bull (Tokyo)1991,39(2):458-464.
55.Kaniwa N,Shameen M,Katori N:The suitable temperature for the evaluation of controlled release products in vitro.Pharmazie 1995,50(2):53-55.
56.Soppimath KS,Aminabhavi TM,Kulkarni AR,Rudzinski WE:Biodegradable polymeric nanoparticles as drug delivery devices.J Control Release 2001,70(1-2):1-20.
57.Oosha E Muller Rh,Davis SS:A continous flow system for in vitro evaluation of drug-loaded biodegradable colloidal carriers.J Pharm Sci 1988,40(6):138-145.
58.杨帆.,张永明.,赵耀明.,李丹涛.,宋风兰.,徐安龙.:干扰素微球的制备及其体外释药性能研究.中国医院药学杂志2005,25(9):807-809.
59.Vert M,Mauduit J,Li S:Biodegradation of PLA/GA polymers:increasing complexity.Biomaterials 1994,15(15):1209-1213.
60.Schakenraad JM,Hardonk MJ,Feijen J,Molenaar 1,Nieuwenhuis P:Enzymatic activity toward poly(L-lactic acid)implants.J Biomed Mater Res 1990,24(5):529-545.
61.lsobe M,Yamazaki Y,Oida S:Bone morphogenetic protein encapsulation with a biodegradable and biocompatible polymer.J Biomed Mater Res 1996,32(10):433-438.
62.Kamei S,Inoue Y:Phagocytosis of microspheres by macrophages.Adv Polym Sci 1990,94(5):107-141.
63.Visscher GE,Pearson JE,Fong JW,Argentieri G J,Robison RL,Maulding HV:Effect of particle size on the in vitro and in vivo degradation rates of poly(DL-lactide-co-glycolide)microcapsules.J Biomed Mater Res 1988,22(8):733-746.
64.Park TG:Degradation of poly(lactic-co-glycolic acid)microspheres:effect of copolymer composition.Biomaterials 1995,16(15):1123-1130.
65.Cadee JA,Brouwer LA,den Otter W,Hennink WE,van Luyn MJ:A comparative biocompatibility study of microspheres based on crosslinked dextran or poly(lactic-co-glycolic)acid after subcutaneous injection in rats.J Biomed Mater Res 2001,56(4):600-609.
66.Shive MS,Anderson JM:Biodegradation and biocompatibility of PLA and PLGA microspheres.Adv Drug Deliv Rev 1997,28(1):5-24.
67.Visscher GE,Robison RL,Maulding HV,Fong JW,Pearson JE,Argentieri G J:Biodegradation of and tissue reaction to 50:50 poly(DL-lactide-co-glycolide)microcapsules.J Biomed Mater Res 1985,19(3):349-365.
68.Visscher GE,Robison RL,Maulding HV,Fong JW,Pearson JE,Argentieri G J:Biodegradation of and tissue reaction to poly(DL-lactide)microcapsules.J Biomed Mater Res 1986,20(5):667-676.
69. Visscher GE, Robison MA, Argentieri GJ: Tissue response to biodegradable injectable microcapsules. J Biomater Appl 1987, 2( 1): 118-131.
70. Yanaguchi K, Anderson JM: Biocompatibility studies of naltrexone sustained release formulations. J Control Release 1992, 19(2):299-314.
71. Grizzi I, Garreau H, Li S, Vert M: Hydrolytic degradation of devices based on poly(DL-lactic acid) size-dependence. Biomaterials 1995, 16(4):305-311.
72. Bostman O: Intense granulomatous in inflammatory lesions associated with absorbable internal fixation devices made of polyglycolide in ankle fractures. Clin Orthop Relat Res 1992, 778(5): 178-199.