离体大鼠肝脏灌流法制备氯吡格雷活性巯基代谢物

Preparation of active thiol metabolite of clopidogrel by isolated rat liver perfusion

目的建立氯吡格雷活性巯基代谢物(CATM)的制备方法,为顺式CATM的合成提供参考。方法以(S)-2-氧氯吡格雷为底物,采用离体大鼠肝脏灌流法和ChromCore 120 C18制备柱制备并分离纯化CATM。采用质谱和核磁共振氢谱鉴定目标产物,对比人体内活性构型(顺式CATM)保留时间确认目标产物中活性构型的占比。结果目标产物的转化率为11.71%,质谱和核磁共振氢谱鉴定目标产物为CATM。CATM的峰2~峰5为4个立体异构体,保留时间分别为21.3、22.3、26.5、27.3 min,峰面积占比分别为7.13%、7.23%、63.52%、14.97%。根据人体内CATM活性构型的保留时间为26.3 min,确定目标产物CATM中活性顺式异构体占比为63.52%。结论本方法成本低、步骤简单,可制备出活性构型占比较高的CATM。

OBJECTIVE To establish the preparation method of clopidogrel active thiol metabolite(CATM),and to provide reference for the synthesis of cis-CATM.METHODS CATM was prepared,separated and purified with isolated rat liver perfusion and ChromCore 120 C18 preparative column,using(S)-2-oxo-clopidogrel as substrate.The target compounds were identified by mass spectrometry and nuclear magnetic resonance spectroscopy.The retention time of the active configuration of CATM in the human body(cis-CATM)were compared to confirm the proportion of active configuration in the target product.RESULTS The conversion rate of the target product was 11.71%.The target products were identified as CATM by MS and 1H-NMR.Peak 2-peak 5 of CATM were four stereoisomers.The retention time of them were 21.3,22.3,26.5,27.3 min.The peak area ratios of them were 7.13%,7.23%,63.52%,14.97%,respectively.Based on that retention time of the active configuration of CATM in human body was 26.3 min,the active cis-stereoisomer in the target product CATM accounted for 63.52%.CONCLUSIONS This method is low-cost,simple,and can prepare CATM with higher active configuration.