A cell-based method for detecting the activity of vitamin K epoxide reductase

摘要：
维生素K环氧化物还原酶(VKOR)是开发抗凝药的重要靶点,其活性测定对鉴定靶向VKOR新型抑制剂具有重要的应用价值.目前测定VKOR活性的常用方法为基于报告基因的方法,是一种间接测定VKOR活性的方法,不能用于鉴定靶向VKOR新型抑制剂.为建立一种快速、直接测定VKOR活性的方法,首先建立了一种基于HPLC检测维生素K(VK)和维生素K环氧化物(VKO)的方法,该方法简便快速,准确性高,重复性好,各检测物色谱峰分离良好.在此基础上,利用GGCX基因敲除的细胞,建立了直接测定细胞内VKOR活性的方法,更加便捷地鉴定靶向VKOR抑制剂,为研发靶向VKOR新型抗凝药提供帮助.

Vitamin K epoxide reductase(VKOR)is an important target for the development of anticoagulant.It is applied with important value to measure the VKOR activity for the identification of new inhibitors targeting VKOR.However,a reporter-gene assay has been widely used to determine VKOR activity indirectly,it could not be used to identify new inhibitors targeting VKOR.In order to establish a method for determination of VKOR activity rapidly and directly,which can be used to identify inhibitors targeting VKOR,here we develop a method for the determination of vitamin K and vitamin K epoxide based on HPLC firstly.The method is simple,rapid,accurate,reproducible to determine vitamin K and vitamin K epoxide,which can be separated well.On the basis of determination of vitamin K and vitamin K epoxide,we use a GGCX gene knockout cells to determine its intracellular VKOR activity directly,which can be used to identify specific VKOR inhibitors easily and quickly.This method will provide technical support for the development of new anticoagulants targeting VKOR.

作者：
徐亚奇 王一伊 李飞 邱稳辉 沈滟 沈国民

Xu Yaqi;Wang Yiyi;Li Fei;Qiu Wenhui;Shen Yan;Shen Guomin(School of Basic Medical Science,Henan University of Science and Technology,Luoyang 471023,China;Henan International Joint Laboratory of Thrombosis and Hemostasis,Henan University of Science and Technology,Luoyang 471023,China)

机构地区：
河南科技大学基础医学院 河南科技大学河南省血栓与止血国际联合实验室

出处：
《betway官方app 学报：自然科学版》 CAS 北大核心 2022年第4期107-111,共5页

Journal of Henan Normal University(Natural Science Edition)

基金：
国家自然科学基金(82170133,81770140) 河南省科技攻关项目(212102310629 212102310877)。

关键词：
维生素K环氧化物还原酶 高效液相色谱 维生素K 维生素K环氧化物 华法林

vitamin K epoxide reductase high performance liquid chromatography vitamin K vitamin K epoxide warfarin

分类号：
R446 [医药卫生—诊断学]

一种基于细胞测定维生素K环氧化物还原酶活性的方法.pdf