绒毛崖豆藤中一种查尔酮提取物的体内外代谢及药代动力学研究

Study on the metabolites and pharmacokinetics of a chalcone extract from Millettia velutina Dunn

采用UPLC-QE-Orbitrap-MS检测方法,通过比较2,5-二甲氧基呋喃[4″,5″:3,4]查耳酮(1)在体外大鼠、小鼠、恒河猴、Beagle犬和人五个种属肝微粒体体系中孵育0 min和60 min的样品,以及比较静脉注射该化合物及空白的C57小鼠的血浆、粪便、尿液样品,研究其在体内外代谢产物。初步推断孵育后体外代谢产物有M1、M2共2种,体内代谢产物M1、M2、M3、M4、M5共5种,并且M5为M1进一步代谢而来,采用化学合成方法制得代谢产物M1。将大鼠随机分组并给药,于设定的系列时间点取血,采用超高速液相色谱质谱联用(UFLC-MS/MS)检测方法,测定1及M1血药浓度,定量研究该化合物及M1在大鼠体内的变化趋势并计算药代参数。用DAS 2.0软件计算药代动力学参数,1的C_(max)=405.96μg/L,Tmax=0.083 h,AUC_(0-t)=190.64μg/(L·h),T_(1/2)=3.74 h,M1的C_(max)=281.291μg/L,T_(max)=0.083 h,AUC_(0-t)=561.30μg/(L·h),T_(1/2)=3.01 h。研究结果表明,2,5-二甲氧基呋喃[4″,5″:3,4]查耳酮在体内代谢过程中主要发生了还原、去甲基、开环、加羟基等反应,其中主要代谢产物为M1,并且M1进一步代谢为M5,本研究为揭示其药理药效作用奠定了物质基础。

The UPLC-QE-Orbitrap-MS assay was used to compare the 2,5-dimethoxyfurano[4″,5″:3,4]chalcone(1)samples cultured in rat,mouse,rhesus monkey,Beagle dog and human liver microsomes for 0 min and 60 min in vitro,and compare the plasma,feces,and urine samples of intravenous injection of the compound and blank C57 mice to study its metabolites in vitro and in vivo.It was preliminarily concluded that there were two kinds of metabolites M1 and M2 in vitro after incubation,and five kinds of metabolites M1,M2,M3,M4 and M5 in vivo.M5 was further metabolized from M1,and the metabolite M1 was obtained by organic synthesis.Ultrafast liquid chromatography-mass spectrometry(UFLC-MS/MS)detection method was used to determine the plasma concentrations of 1 and M1.The change trend of the compound and M1 in rats was quantitatively studied and the pharmacokinetic parameters were calculated.Rats were randomly divided into groups and given drugs,and blood was collected at a set series of time points.The plasma concentrations of 1 and M1 were determined.The pharmacokinetic parameters were calculated by DAS2.0 software.The C_(max) of 1 was 405.96μg/L,T_(max)=0.083 h,AUC_(0-t)=190.64μg/(L·h),T_(1/2)=3.74 h.The C_(max) of M1 was 281.291μg/L,T_(max)=0.083 h,AUC_(0-t)=561.30μg/(L·h),T_(1/2)=3.01 h.These results showed that 2,5-dimethoxyfuran[4″,5″:3,4]chalcone undergoes reduction,demethylation,ring opening,and hydroxylation reactions during the metabolism of the body.The main metabolite is M1,and M1 is further metabolized to M5.This study laid a material foundation for revealing its pharmacological effects.