摘要
目的通过凝血酶亲和色谱前沿分析计算阿加曲班、隐丹参酮、二氢丹参酮Ⅰ及丹参酮ⅡA四种化合物与凝血酶之间的平衡解离常数(KD)。方法制备凝血酶亲和色谱柱(10 mm×2.0 mm,5μm),4种化合物浓度由低到高依次通过色谱柱,制作突破曲线,记录拐点时间。根据前沿色谱理论,以1/[L]m对1/mL作图,回归曲线斜率和截距之比即为KD。结果阿加曲班的平衡解离常数为(6.95±0.61)×10^(-9)mol·L^(-1),与文献报道的数据5.0 nmol·L^(-1)接近。隐丹参酮、二氢丹参酮Ⅰ及丹参酮ⅡA的平衡解离常数分别为(13.49±0.92)×10^(-9)、(3.24±0.32)×10^(-9)和(2.88±0.31)×10^(-9)mol·L^(-1)。亲和强度顺序为:隐丹参酮<阿加曲班<二氢丹参酮Ⅰ<丹参酮ⅡA。结论本法可用于药物与凝血酶间平衡解离常数KD值的计算。
Objective To calculate the equilibrium dissociation constant(KD)between argatroban,cryptotanshinone,dihydrotanshinone Ⅰ and tanshinone ⅡA through the frontier analysis of thrombin affinity chromatography.Methods A thrombin affinity chromatography column(10 mm×2.0 mm,5μm)was prepared.Each compound was passed through the column from low to high concentration.A breakthrough curve was made and the turning point time was recorded.According to the frontier chromatography theory,the model was plotted with 1/[L]m versus 1/mL.The ratio of the slope to the intercept of the regression curve is KD.Results The KD of argatroban was(6.95±0.61)×10^(-9) mol·L^(-1),which was close to the data 5.0 nmol·L^(-1)in the literature.The KD values of cryptotanshinone,dihydrotanshinone Ⅰ and tanshinone ⅡA were(13.49±0.92)×10^(-9),(3.24±0.32)×10^(-9) and(2.88±0.31)×10^(-9) mol·L^(-1),respectively.The order of affinity strength is:cryptotanshinone<argatroban<dihydrotanshinone Ⅰ<tanshinone ⅡA.Conclusion This method can be used to calculate the equilibrium dissociation constant between the drugs and thrombin.
作者
王欣
邓子杰
胥丹
郭东男
乔艳茹
王嗣岑
侯晓芳
WANG Xin;DENG Zijie;XU Dan;GUO Dongnan;QIAO Yanru;WANG Sicen;HOUXiaofang(Department of Pharmacy,Xi'an No.1 Hospital,Xi'an 710002,China;School of Pharmacy,Xi'an Jiaotong University,Xi'an 710061,China)
出处
《沈阳药科大学学报》
CAS
CSCD
北大核心
2021年第10期1013-1018,共6页
Journal of Shenyang Pharmaceutical University
基金
陕西省科技厅面上项目(2021JM-039)
中国博士后科学基金面上项目(2018M643681)。
关键词
凝血酶
亲和色谱
前沿分析法
平衡解离常数
thrombin
affinity chromatography
the frontier analysis
the equilibrium dissociation constant(KD)
