目的 建立測定肝微粒體孵育體系中訶子酸濃度的方法，并比較其在人、犬、猴、小鼠、大鼠肝微粒體中的Ⅰ相、Ⅱ相代謝穩(wěn)定性及種屬差異，確定其在人肝微粒中的代謝表型。方法 將訶子酸與不同種屬肝微粒體共同孵育，應用UPLCMS/MS檢測孵育液中訶子酸的含量，考察其代謝穩(wěn)定性及體外動力學參數(shù)。采用化學抑制劑法確定其在人肝微粒中的代謝表型。將訶子酸與各CYP450同工酶CYP1A2、CYP2A6、CYP2C9、CYP2C19、CYP2D6、CYP2E1和CYP3A4的特異性抑制劑（α-萘黃酮、香豆素、磺胺苯吡唑、噻氯匹定、奎尼丁、二乙基二硫代氨基甲酸鈉、酮康唑）共同孵育，確定其代謝酶表型。結(jié)果 訶子酸在Ⅰ相、Ⅱ相孵育體系中均可代謝，在Ⅰ相代謝中，犬肝微粒體孵育與人最為相似，半衰期（t_1/2）分別為115.50 min和121.58 min；Ⅱ相代謝中5個種屬代謝穩(wěn)定性均中等，其中猴與人肝微粒體代謝趨勢最為相近。訶子酸在人肝微粒中的代謝是由多種CYP酶共同介導的，其中CYP2C9、CYP2E1和CYP3A4是主要的同工酶。結(jié)論 建立的UPLCMS/MS方法簡便、快速、專屬性強、靈敏性高，可用于肝微粒體孵育體系中訶子酸濃度的測定及體外代謝的研究。訶子酸在人、犬、猴、小鼠、大鼠肝微粒體中代謝存在一定種屬差異，且其代謝過程與多種CYP酶相關。

Objective To establish a method for the concentration determination of chebulinic acid (CA) in liver microsomes, and to compare its phase I and phase II metabolic stability and species differences in human, dogs, monkeys, mice and rats liver microsomes, to predict the metabolic phenotypes of CA in human liver microsomes. Methods The remaining concentrations of CA incubated in various species of liver microsomes were determined to evaluate their metabolic stability and in vitro pharmacokinetic parameters. Selective chemical inhibition was used to predict the metabolic phenotypes of CA in human liver microsomes. CA was incubated with specific inhibitors of CYP1A2, CYP2A6, CYP2C9, CYP2C19, CYP2D6, CYP2E1 and CYP3A4 isoforms (α-naphthoflavone, coumarin, sulfaphenazolum, ticlopidine, quinidine, sodium diethyldithiocarbamatre, ketoconazole) to determine the phenotypes of metabolic enzymes. Results CA can be metabolized in phase I and phase II incubation systems. In phase I metabolism, the metabolic properties of dogs and human liver microsomes were the most resembles, and the half-lives are 115.50 min and 121.58 min, respectively. Metabolic stability in phase Ⅱ metabolism is moderate, and dogs, monkeys and human liver microsomes have the similar metabolic characteristic. CA is metabolized by a variety of enzymes in human liver microsomes, among which CYP2C9, CYP2E1 and CYP3A4 are the primary isoenzymes. Conclusion A UPLC-MS/MS method was established to determine the concentration of CA, and it is simple, rapid and specific. It can be used for the concentration determination of CA in liver microsomes incubation system. The metabolic characteristics of the compound are different among human, dogs, monkeys, mice and rats liver microsomes. It is metabolized by a variety of enzymes in human liver microsomes.

R285.5

中央高校基本科研業(yè)務費專項資助項目(3332021064);國家重點研發(fā)計劃資助項目(2018YFC1708203);內(nèi)蒙古自治區(qū)科技重大專項資助(2019ZD004)