摘要
目的 发现并证明大分子杂质是影响注射用聚山梨酯80安全性的因素,测定去大分子聚山梨酯80的主要物理常数。方法理论分子量为1310 Da的注射用聚山梨酯80原液用超滤膜得到组分A(>100 kDa)、组分B(10~100 kDa)和组分C(<10 kDa)。各组分冷冻干燥后获得各组分在原液中的比例。各组分用生理盐水配置成10%浓度,按照《中国药典》2015年版进行豚鼠过敏和类过敏反应检查,同时用昆明种小鼠进行静脉注射的5 d毒性检查,并观察5 d恢复期。按照《中国药典》方法检测各组分的物理常数。结果分子量大于10 kDa的组分占85%以上,聚山梨酯80原液、组分A和组分B能诱发过敏和类过敏反应,而组分C反应阴性。组分A和B能在5 d恢复期减慢体重恢复;根据器官指数变化,聚山梨酯80原液能导致小鼠肝脏、肾脏、脾脏和肺轻度损伤,5 d后仍不能完全恢复;其中肝肾损伤得到了显微病理支持。其中组分C对各器官的影响最小,且容易恢复。去大分子后(组分C),其增溶能力、粘度、密度、吸收光谱、pH和颗粒大小得到了不同程度的改善。结论注射用聚山梨酯80中含有大量的大分子杂质,这些杂质是导致器官毒性、过敏和类过敏反应的重要因素,去除大分子杂质能提高其安全性。
Objective The major aim of the present study were to prove that macromolecular impurities in polysorbate 80 for injection are a risk factor,and determine the main physical indexes of polysorbate 80 free of macromolecular substances.Methods The original polysorbate 80 for injection,with a theoretical molecular weight of 1310 Da,was treated with ultra-filters to obtain component A(>100 kDa),B(10~100 kDa),and C(<10 kDa).The components were lyophilized to obtain their contents.Each component was prepared with normal saline at a concentration of 10%,anaphylactic and anaphylactoid reactions on guinea pigs that may be caused by the original polysorbate 80 and its components were recorded according to Chinese Pharmacopoeia of 2015th edition.Kunming mice were intravenously injected with them for 5 days'administration and 5 days’recovery to observe their organic toxicity.Results The proportion of substances with a molecular weight above 10 kDa in the original polysorbate 80 was above 85%.The original polysorbate 80 and its component A and B resulted in positive reactions in both anaphylactic and anaphylactoid guinea pigs,while component C didn't.In Kunming mice,component A and B caused an obvious retard of body weight increase in the 5 days’recovery.The original polysorbate 80 and its components caused mild but significant toxicity in the liver,kidney,spleen,and lung at organ indexes both in 5 days administration and recovery.Their mild toxicity on the liver and kidney was verified by histological assay.However,toxicity caused by component C was much milder and more recoverable.The physicochemical constants of component C,including solubilization capacity,viscosity,density,absorption spectrum,pH,and particle size were improved.Conclusion All the results suggested that the main substances in polysorbate 80 for injection are macromolecular impurities that are an important risk factor to cause anaphylactoid reactions and organic toxicity,and macromolecular impurity removal improved its safety and physicochemical constants.
作者
夏恒
段金连
李月
邱玲
韩依伦
陈欢
黎勇坤
段为钢
XIA Heng;DUAN Jinlian;LI Yue;QIU Ling;HAN Yilun;CHEN Huan;LI Yongkun;DUAN Weigang(Yunnan Provincial Key Laboratory of Molecular Biology for Sinomedicine,Yunnan University of Chinese Medicine,Kunming 650500,China)
出处
《云南中医学院学报》
2021年第1期1-16,共16页
Journal of Yunnan University of Traditional Chinese Medicine
基金
国家自然科学基金(81560645)
云南省中医药联合专项重点项目(2018FF001-002)。
关键词
注射用聚山梨酯80
大分子杂质
类过敏反应
器官毒性
临界胶束浓度
增溶
polysorbate 80 for injection
macromolecular impurities
anaphylactoid reactions
organic toxicity
critical micelle concentration
solubilization
