卡式瓶多剂量重组人干扰素α2b注射液与冻干制剂的比对研究及优势分析

Comparative study and analysis of advantages of multi-dose recombinant human interferonα2b injection in cartridges and freeze-dried preparation

目的通过剂型比对探讨卡式瓶多剂量重组人干扰素α2b注射液的剂型优势。方法将长春生物制品研究所有限责任公司在研卡式瓶多剂量重组人干扰素α2b注射液与同公司市售注射用重组人干扰素α2b各3批,同时置(25±2)℃避光条件下储存,0、1、2、3和6个月取样,亲和层析前处理浓缩后样品,分别进行非还原型SDSPAGE、SEC-HPLC、质谱肽图、质谱二硫键分析,比对两种剂型中蛋白聚合、降解、修饰产物种类及变化趋势;未经处理样品直接进行生物学活性检测,比对两种剂型有效性指标的变化趋势。结果SDS-PAGE和SEC-HPLC分析结果相符,两种剂型在加速比对研究周期内均可见蛋白二聚体产生,冻干制剂可见多种蛋白多聚体产生,液体制剂仅见单一种类的蛋白多聚体产生,冻干制剂的电泳纯度和色谱纯度下降趋势较液体制剂明显;质谱分析结果显示,两种剂型均产生了乙酰化修饰产物、氧化修饰产物和二硫键错配产物,两种剂型内各种修饰产物的变化趋势趋于一致;两种剂型生物学活性无明显下降趋势,稳定程度相似。结论长春生物制品研究所有限责任公司卡式瓶多剂量重组人干扰素α2b注射液(25±2)℃存放6个月内聚合产物少,蛋白纯度下降速度慢,生物学活性稳定无降低趋势,极具剂型优势。

Objective To investigate the advantages of multi-dose recombinant human interferon(rhIFN)α2 b injection in cartridges by the comparison of dosage forms.Methods Three batches of multi-dose rhIFNα2 b injection in cartridges and three batches of commercial freeze-dried rhIFNα2 b for injection,either manufactured by Changchun Institute of Biological Products Co.,Ltd.,were stored at(25±2)℃,protected from light,from which samples were taken 0,1,2,3 and 6 months after storage,concentrated,pretreated by affinity chromatography,and then analyzed by SDS-PAGE,SECHPLC,mass spectrometry peptide mapping and mass spectrometry disulfide bond assay.The protein polymerization,degradation as well as kind and change trend of modification products of the two dosage forms were compared.The untreated samples were analyzed for biological activity to compared the change trends of effectiveness indexes of the two dosage forms.Results SDS-PAGE and SEC-HPLC showed visible protein dimers in both the dosage forms in the accelerating comparison research cycle.A variety of protein polymer was observed in freeze-dried preparation,while only a single kind of protein polymer in liquid preparation.The electrophoretic and chromatographic purities of freeze-dried preparation decreased more obviously than those of liquid preparation.Mass spectrometry showed that acetylated modified products,oxidized modified products and disulfide bond mismatch products were produced in both the dosage forms,of which the change trends were in agreement.However,the biological activities of the two dosage forms showed no significant decrease,while the stabilities were similar.Conclusion Less polymers were produced in the multi-dose rhIFNα2 b injection in cartridges,manufactured by Changchun Institute of Biological Products Co.,Ltd.,within 6 months after storage at(25±2)℃,while the protein purity decreased slowly,and the biological activity showed no decreasing trend,indicating the advantage of dosage form.