重组大熊猫IFN-γ聚氰基丙烯酸正丁酯纳米球的制备与评价

Preparation and Evaluation of Recombinant Interferon-gamma Nanosphere Loaded in Polybutylcyanoacrylate in Giant Panda(Ailuropoda melanoleuca)

为了制备并评价大熊猫(Ailuropoda melanoleuca)重组干扰素γ(IFN-γ)聚氰基丙烯酸正丁酯(PBCA)纳米球,本研究利用蛋白质原核表达技术,获得了重组大熊猫IFN-γ蛋白,然后以PBCA为载药材料,采用乳化聚合法制备了大熊猫干扰素γ纳米微球(IFNγ-PBCA-NS),最后借助感染大熊猫流感病毒A/Panda/Sichuan/01/2011(H1N1)的昆明小鼠(Mus musculus)模型,通过灌胃和皮下注射药物初步评价了IFNγ-PBCA-NS的药效。结果表明,大熊猫IFN-γ的蛋白分子量约为33.5 ku,所制备的大熊猫IFNγ-PBCA-NS外观规整,粒径在50~200 nm之间,跨距0.55,大小较均匀,包封率为56.7%,载药量为0.86%。灌胃和注射两种给药途径中,IFNγ-PBCA-NS组小鼠的生命延长率均显著高于IFN-γ组(P<0.05或P<0.01)。显示IFNγ-PBCA-NS在小鼠体内有更好的缓释和抗病毒作用,可为进一步研究制备大熊猫多肽类药物微粒提供参考。

To prepare and investigate the recombinant Interferon-gamma（IFN-γ） nanosphere loaded in polybutylcyanoacrylate（PBCA） in Giant Panda（Ailuropoda melanoleuca）,we firstly obtained the recombinant IFN-γ protein through prokaryotic protein expression technology,and then,by choosing PBCA as the drug delivery material,prepared the INF-γ PBCA nanosphere（IFNγ-PBCA-NS） using an emulsion polymerization method.Further more,the Kunming Mice（Mus musculus）,which were infected with giant panda influenza virus A/Panda/Sichuan/01/2011（H1N1）,were used as models to test the efficacy of IFNγ-PBCA-NS compared to that of IFNγ after intragastric administration and subcutaneous injection of drugs.The results demonstrated that the molecular weight of INF-γ protein was about 33.5 ku,and the IFNγ-PBCA-NS was globular in appearance,with uniform particle size.The diameters ranged from 50 to 200 nm,the span was 0.55,the encapsulation efficiency of the particles was 56.7% and the drug loading was 0.86%.Moreover,the prolonged life rate of mice in the IFNγ-PBCA-NS group was significantly higher than that of the IFNγ group in both intragastric administration and subcutaneous injection approaches（P〈0.05 or P 〈0.01）.In summary,IFNγ-PBCA-NS showed better sustained release and antiviral activity in mice,which could provide references for further research on the preparation of the Giant Panda peptide drug microparticles.