海南崖豆藤提取物HN-1的体内外代谢研究

Investigation of metabolism of HN-1 isolated from Millettia pachyloba in vivo and in vitro

应用体外肝微粒体孵育体系,采用超快速液相色谱质谱联用(UFLC-MS/MS)检测方法,研究海南崖豆藤抗炎活性成分6-methoxy-8,8-dimethyl-3-(2,4,5-trimethoxyphenyl)-4H,8H-pyrano[2,3-f]chromen-4-one(HN-1)在大鼠、小鼠、恒河猴、Beagle犬和人5个种属肝微粒体中的代谢稳定性,比较代谢种属间的差异;采用化学抑制剂法确定其在人肝微粒体中的代谢表型。采用UPLC-Q-TOF-MS/MS检测方法,通过比较体外孵育0,60 min的样品,初步推断各种属体外代谢产物;比较粪便、尿液、血浆空白和给药后的生物样品,推测HN-1在SD大鼠体内的代谢产物。结果表明HN-1在各种属肝微粒体中的代谢均较为稳定,其中犬与人的肝微粒体代谢性质最相近;在人肝微粒体中主要经CYP1A1,CYP2D6,CYP3A4 3种同工酶催化代谢;初步推测出HN-1在体内体外的代谢产物,其中包括3个体外代谢产物和5个体内代谢产物,研究结果为HN-1进一步的药理药效研究奠定基础。

Ultra-fast performance liquid chromatography-mass spectrometry(UFLC-MS/MS)was used to study the anti-inflammatory active ingredient of Millettia pachyloba,6-methoxy-8,8-dimethyl-3-(2,4,5-trimethoxyphenyl)-4 H,8 H-pyrano[2,3-f]chromen-4-one(HN-1),in liver microsomes of rats,mice,rhesus monkeys,Beagle dogs and humans metabolic stability,and compare the metabolic differences between different species.The metabolic phenotype in human liver microsomes was determined by chemical inhibitor method.Using UPLC-Q-TOF-MS/MS detection method,the in vitro metabolites of various liver microsomes were preliminarily inferred by comparing the samples incubated for 0 min and 60 min in vitro.The metabolites of HN-1 in SD rats were presumed by comparing feces,urine,plasma blanks and samples after administration.The results showed that the metabolism of HN-1 in various liver microsomes was stable,and the metabolic properties of dog and human liver microsomes were the closest.It is mainly catabolized by CYP1 A1,CYP2 D6 and CYP3 A4 isoenzymes in human liver microsomes.The metabolites of HN-1 in vitro and in vivo,including 3 in vitro metabolites and5 in vivo metabolites,were preliminarily estimated.The results laid the foundation for further pharmacological studies of HN-1.