CAJ | 학술논문

目的制备葡激酶突变体(K35R,DGR)的聚乳酸-羟基乙酸(PLGA)微球,使其在包封和释放过程中都能保持活性.方法使用复乳溶剂挥发法制备DGR的PLGA微球,研究了搅拌速度、PLGA浓度、内水相和外水相中的添加剂对蛋白包封率以及微球性质的影响,并进行了DGR微球的体外和体内释放试验.结果 2%聚乙烯醇可以有效抑制超声乳化时DGR在水/二氯甲烷界面上的变性,将DGR的活性回收率从16%提高到几乎100%.在外水相中加入NaCl可以显著提高蛋白包封率,同时对微球的粒径分布和表面形态也产生了重要影响.DGR微球的体外释放呈现两个时相,15 d释放大约DGR总活性的50%.DGR微球在体内持续释放5 d.结论制备的PLGA微球,DGR包封率高,稳定性较好,是DGR的良好载药系统.
목적제비포격매돌변체(K35R,DGR)적취유산-간기을산(PLGA)미구,사기재포봉화석방과정중도능보지활성.방법사용복유용제휘발법제비DGR적PLGA미구,연구료교반속도、PLGA농도、내수상화외수상중적첨가제대단백포봉솔이급미구성질적영향,병진행료DGR미구적체외화체내석방시험.결과 2%취을희순가이유효억제초성유화시DGR재수/이록갑완계면상적변성,장DGR적활성회수솔종16%제고도궤호100%.재외수상중가입NaCl가이현저제고단백포봉솔,동시대미구적립경분포화표면형태야산생료중요영향.DGR미구적체외석방정현량개시상,15 d석방대약DGR총활성적50%.DGR미구재체내지속석방5 d.결론제비적PLGA미구,DGR포봉솔고,은정성교호,시DGR적량호재약계통.
Aim To produce poly (lactic-co-glycolic acid) (PLGA) microspheres, containing a staphylokinase variant (K35R, DGR) with reduced immunogenecity and antiplatclet aggregation activities,which allowed the preservation of protein stability during both particle processing and drug release.Methods DGR-loaded microspheres were fabricated using a double emulsion-solvent evaporation technique. The effects of preparative parameters, such as stirring rate, polymer concentration, and the excipients of both internal and external aqueous phase (W2 ), on DGR encapsulation efficiency and microsphere characteristics were investigated. In vitro and in vivo release of DGR were conducted and the cause for instability of DGR during release was also investigated. Results Moderate ultrasonic treatment of aqueous DGR/dichloromethane mixtures caused approximately. Eighty four per cent DGR denaturation.However, the activity recovery of DGR almost amounted to 100% when 2% polyvinyl alcohol (PVA) was added into the aqueous phase. It was found that NaCl in the external water phase significantly increased DGR encapsulation efficiency. Furthermore, NaCl in the external water phase played a role in determining size and surface morphology of microsphere. In vitro release test showed a burst release of DGR from microspheres, followed by sustained release of 50% total activity over 15 days. In vivo experiments showed that DGR released from microspheres sustained 5 days. Denaturation of DGR within microspheres might be resulted from acidic microclimate. Conclusion The stability of DGR was effectively protected during microencapsulation and a relatively high encapsulation efficiency of DGR was obtained. PLGA microspheres could be an effective carrier for DGR.