摘要
目的 对注射用重组白细胞抑制因子和水蛭肽嵌合蛋白(human recombinant neutrophil inhibitory factor and hirulog hybrid,TNHH)的处方及工艺进行筛选和优化,并考察其稳定性。方法 以单因素试验结果为依据,pH范围、甘露醇用量、聚维酮K30用量为自变量,高分子蛋白含量为响应值,采用CCF响应曲面设计试验,分析各自变量及其交互作用对注射用重组白细胞抑制因子和水蛭肽嵌合蛋白中高分子蛋白含量的影响,筛选出最优处方。为了方便操作,对最优处方调整后制备中试规模3批样品,分别置于40℃、相对湿度(relative humidity,RH)75%条件下2、4周,2~8℃条件下3、6个月,取样,检测外观、pH、反相高效液相色谱(reversed phase-high performance liquid chromatography,RP-HPLC)纯度和体积排阻高效液相色谱(size exclusion chromatography-high performance liquid chromatography,SECHPLC)纯度,验证该处方和工艺的稳定性。结果 筛选出的最优处方为:pH 4.982 6,甘露醇用量7.986 4%,聚维酮K30用量1.902 7%,最终调整为pH 5.0,甘露醇用量8.0%,聚维酮K30用量2.0%。采用优化处方和工艺制备的注射用重组白细胞抑制因子和水蛭肽嵌合蛋白制剂质量稳定,符合临床用药要求。结论 优选的注射用重组白细胞抑制因子和水蛭肽嵌合蛋白制剂处方工艺合理,适合工业化生产。
Objective To screen and optimize the formulation and technology of human recombinant neutrophil inhibitory factor and hirulog hybrid(TNHH)for injection,and investigate its stability.Methods Based on the results of the single factor experiment,with the pH range,mannitol dosage and povidone K30 dosage as independent variables,and the content of high molecular protein as response value,the response surface design(CCF)test was used to analyze the effects of the respective variables and their interaction on the content of high molecular protein in TNHH for injection to screen out the optimal formulation. In order to facilitate the operation,the optimal formulation was adjusted to prepare three batches of samples in pilot scale,which were placed at 40 ℃,75% relative humidity(RH)for 2,4 weeks and 2 ~ 8 ℃ for 3,6 months,respectively. The samples were taken and the appearance,pH,purity of reversed phase-high performance liquid chromatography(RP-HPLC)and purity of size exclusion chromatography-high performance liquid chromatography(SECHPLC)were detected to verify the stability of this formulation and process.Results The optimal formulation was pH 4. 982 6,mannitol 7. 986 4% and povidone K30 1. 902 7%,which was finally adjusted to pH 5. 0,mannitol 8. 0% and povidone K302. 0%. The TNHH preparation for injection prepared by the optimized prescription and process were stable in quality and met the clinical medication requirements.Conclusion The optimum formulation of TNHH preparation for injection is reasonable in the process and suitable for industrial production.
作者
李娜
王轲
张培彪
刘忠
张贵民
LI Na;WANG Ke;ZHANG Peibiao;LIU Zhong;ZHANG Guimin(Shandong Engineering Laboratory of Protein Drugs,Shandong Newtimes Pharmaceutical CO.,Ltd.,Linyi 273400,Shandong Province,China;不详)
出处
《中国生物制品学杂志》
CAS
CSCD
北大核心
2023年第8期967-972,979,共7页
Chinese Journal of Biologicals
基金
山东省重点研发计划(泰山产业领军人才工程)(tscy20200329)
省新旧动能转换重大工程重大课题攻关项目(临发改动能[2020]235号)。
关键词
重组白细胞抑制因子和水蛭肽嵌合蛋白
冻干粉
响应曲面设计
Human recombinant neutrophil inhibitory factor and hirulog hybrid
Lyophilized powder
Response surface methodoligy(RSM)