聚乙二醇衍生物修饰的IL-12制备及体外评价  

Preparation and in vitro evaluation of polyethylene glycol derivatives modified IL-12

作　　者：刘艺 邹镇阳 李宗艳 韩峰[1] 乔天奎 王建刚 LIU Yi;ZOU Zhenyang;LI Zongyan;HAN Feng;QIAO Tiankui;WANG Jiangang(School of Medicine and Pharmacy,Ocean University of China,Qingdao 266000,China;Kanglitai Biomedical(Qingdao)Co.,LTD,Qingdao 266000,China)

机构地区：[1]中国海洋大学医药学院,青岛266000 [2]康立泰生物医药(青岛)有限公司,青岛266000

出　　处：《中国免疫学杂志》2024年第7期1495-1500,1506,共7页Chinese Journal of Immunology

基　　金：国家科技重大专项“重大新药创制”(2017ZX09303017);山东省重大科技创新工程项目(NO2018CXGC1404)。

摘　　要：目的:制备聚乙二醇衍生物修饰的IL-12,确定其修饰位点,并在稳定性、血象恢复、抗肿瘤等多方面对其进行体外评价。方法:使用聚乙二醇丙醛、马来酰亚胺衍生物修饰IL-12,超高效液相色谱(UPLC)确定两种不同衍生物的修饰位点;使用重组人IL-12、PEG-IL-12刺激NK92细胞,试剂盒评估IFN-γ活性及细胞毒性;刺激CD34+评估其血象恢复潜力;刺激NK细胞杀伤活力评估其在提高免疫力、抗肿瘤方面的效果。结果:与重组人IL-12相比,PEG-IL-12具有更高的稳定性、血象恢复潜能和抗肿瘤效果。结论:PEG-IL-12可有效克服重组人IL-12的诸多缺点,显著提高IL-12在临床试验中广泛应用的可能。Objective:To prepare IL-12 modified with polyethylene glycol derivatives and determine its modification sites,and to evaluate it in vitro in terms of stability,hemogram recovery,anti-tumor effects and other aspects.Methods:IL-12 was modified with polyethylene glycol propionaldehyde and maleimide imide derivatives,and modified sites of two different derivatives were determined by ultra performance liquid chromatography(UPLC).Recombinant human IL-12 and PEG-IL-12 were used to stimulate NK92 cells,activity and cytotoxicity of IFN-γwere evaluated by kits.CD34+was stimulated to evaluate its blood picture recovery potential.NK cell killing was stimulated to evaluate its effectiveness in improving immunity and anti-tumor effects.Results:Compared with recombinant human IL-12,PEG-IL-12 had higher stability,blood picture recovery potential and anti-tumor effects.Conclusion:PEGIL-12 can effectively overcome many drawbacks of recombinant human IL-12,greatly improving possibility of its widespread application in clinical trials.

关 键 词：聚乙二醇 IL-12 血象恢复 抗肿瘤 

分 类 号：R392.1[医药卫生—免疫学]