重组人凝血酶体外药学及体内药效学的研究  

Research of in vitro pharmacy and in vivo pharmacodynamics of recombinant human thrombin

作　　者：徐文峰[1] 徐硕[1] 张闪闪 邝咏梅[1] 金鹏飞[1] XU Wen-feng;XU Shuo;ZHANG Shan-shan;KUANG Yong-mei;JIN Peng-fei(Department of Pharmacy,Beijing Key Laboratory of Assessment of Clinical Drugs Risk and Individual Application(Beijing Hospital),Beijing Hospital/National Center of Gerontology/Institute of Geriatric Medicine,Chinese Academy of Medical Sciences,Beijing 100730,China)

机构地区：[1]北京医院、国家老年医学中心、中国医学科学院老年医学研究院药学部、北京市药物临床风险与个体化应用评价重点实验室(北京医院),北京100730

出　　处：《中国临床药理学杂志》2024年第23期3420-3423,共4页The Chinese Journal of Clinical Pharmacology

基　　金：首都卫生发展科研专项重点攻关基金资助项目(首发-2024-1-4052)。

摘　　要：目的研究重组人凝血酶的体外药学特性(蛋白质含量及纯度、生物学活性)和体内药效学(促凝血作用)。方法用福林酚法测定重组人凝血酶中蛋白质的含量,以十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法对重组人凝血酶中蛋白质的纯度进行测定,用人凝血酶活性检查法对重组人凝血酶的生物学活性进行测定。将18只雄性新西兰兔用Pristima系统按体质量随机分为空白组(0.9%NaCl溶液)、低剂量实验组(200 U·mL^(-1)重组人凝血酶)、高剂量实验组(1000 U·mL^(-1)重组人凝血酶),每组6只。以兔肝损伤模型考察重组人凝血酶的促凝血作用。结果重组人凝血酶的蛋白质含量为每支(1.980±0.024)mg,生物学活性为每支(5764.3±197.7)U,生物学比活性为(2911.2±99.8)U·mg^(-1),其目的蛋白(非还原电泳35 kD处)条带单一,杂蛋白较少。低、高剂量实验组与空白组相比,可以使兔肝创面的平均凝血时间分别缩短57.0%和71.8%,平均失血量分别减少87.7%和91.9%。结论重组人凝血酶具有较高的蛋白质纯度和生物学比活性,体内促凝血作用显著。Objective To investigate the in vitro pharmaceutical properties(protein content and purity,biological activity)and in vivo pharmacodynamics(procoagulant activity)of recombinant human thrombin.Methods The content of protein in recombinant human thrombin was determined by folin-reagent method.Sodium dodecyl sulphate-polyacrylamide gel electrophoresis was used to determine its purity.Human thrombin activity test was applied to detect its biological activity.Eighteen male New Zealand rabbits were randomly divided according to body mass into blank group(0.9%NaCl solution),experimental-L group(200 U·mL^(-1) recombinant human thrombin)and experimental-H group(1000 U·mL^(-1) recombinant human thrombin),6 rabbits in each group.The procoagulant effects of recombinant human thrombin were investigated in the rabbit liver injury model.Results The protein content of recombinant human thrombin was(1.980±0.024)mg per container,its biological activity was(5764.3±197.7)U per container and its biological specific activity was(2911.2±99.8)U·mg^(-1).The target protein(35 kD in non-reductive electrophoresis)has a single band and few miscellaneous proteins were found on the gel.Compared with the blank group,the me an hemostasis time and mean blood loss weight of rabbit liver wound were reduced by 57.0%and 87.7%in the experimental-L group,which in the experimental-H group were reduced by 71.8%and 91.9%.Conclusion Recombinant human thrombin has high protein purity and specific biological activity,and has significant procoagulant activity in vivo.

关 键 词：重组人凝血酶 蛋白质含量 蛋白质纯度 生物学活性 促凝血作用 

分 类 号：R973.2[医药卫生—药品]