粒细胞-巨噬细胞集落刺激因子缓释微球的研制  

Preparation of Sustained-release Microspheres Loaded with Granulocyte-macrophage Colony-stimulating Factor

作　　者：于如月 邹茜茜 朱琳[1] 吴飞[1] 金拓[1] YU Ruyue;ZOU Xixi;ZHU Lin;WU Fei;JIN Tuo(School of Pharmacy,Shanghai Jiao Tong University,Shanghai 200240)

机构地区：[1]上海交通大学药学院,上海200240

出　　处：《中国医药工业杂志》2021年第12期1615-1620,1621,共7页Chinese Journal of Pharmaceuticals

摘　　要：粒细胞-巨噬细胞集落刺激因子(granulocyte-macrophage colony-stimulating factor,GM-CSF)存在半衰期短(约1 h)、易失活和不易透过胃肠道黏膜的特点,几乎不能口服,其注射剂也需要每日多次给药,患者顺应性较差。聚乙二醇(PEG)化、蛋白质的脂化及常规微球制备方法都对蛋白构象和生物活性有一定破坏,同时存在严重突释和蛋白聚集体形成导致的不完全释放问题。本研究选择GM-CSF为模型蛋白,通过本实验室发明的双水相冷冻相分离方法首先将蛋白制备成葡聚糖(DEX)颗粒预制剂,进而通过s/o/w法制备了DEX颗粒复合PLGA微球。担载GM-CSF的DEX颗粒制成的PLGA微球在d1突释载药量的19%~23%,40 d内连续缓慢释放至载药量的83%~90%,释出的GM-CSF对人红系白血病(TF-1)细胞的增殖活性保持在90%以上。用w/o/w复乳法制得的担载GM-CSF的PLGA微球在d1突释载药量的30%,23 d内缓慢释放至载药量的53%,前6 d释出的GM-CSF的细胞增殖活性为60%,之后下降至50%。2种方法制备的微球的体外释放曲线证明PLGA微球本身在释放过程中可平稳缓释GM-CSF,GM-CSF-DEX颗粒在微球体系中可有效保护GM-CSF的生物活性,同时实现4周以上的平稳缓释。Granulocyte-macrophage colony-stimulating factor(GM-CSF) cannot be administrated by oral route and requires multiple daily injections to achieve therapeutic concentration,which lead to poor compliance.This can be attributed to its short half-life(about 1 h),poor stability and permeability across gastrointestinal mucosa.However,proteins suffer partially deactivation and conformation damage by PEGylation,lipidation of proteins,and conventional preparation methods for microspheres.Otherwise,burst release and incomplete release by aggregations of proteins are impossible to ignore.According to difficulties discussed above,the PLGA microspheres loaded with GM-CSF-dextran(DEX) particles was prepared by a stable aqueous-aqueous "emulsion" followed by lyophilization and then encapsulated into the PLGA microspheres by solid-in-oil-in-water(s/o/w) emulsion method.The burst release on day 1 was 19%-23%,and 83%-90% of total drug loading was released within 40 days.Meanwhile,the preserved bioactivity of GM-CSF determined by human erythroleukemia(TF-1) cell proliferation was more than 90%.The PLGA microspheres of control group was prepared by water-in-oil-in-water(w/o/w) emulsion method.The burst release on day 1 was 30%,and only 53% of total drug loading was released within 23 days.The preserved bioactivity of GM-CSF was 60% within the first 6 days and then dropped to 50%.The PLGA microspheres loaded with GM-CSF-DEX particles was demonstrated to be a feasible strategy to achieve sustained release of GM-CSF for up to 4 weeks and the DEX particles could protect bioactivity of GM-CSF during both the preparation process and the release period.

关 键 词：粒细胞-巨噬细胞集落刺激因子 冷冻相分离 缓释微球 蛋白生物活性 

分 类 号：R944.9[医药卫生—药剂学]