英菲格拉替尼及其活性代谢产物对大鼠肝微粒体中CYPs和UGTs的抑制作用

Inhibitory effects of infigratinib and its pharmacologically active metabolites on CYPs and UGTs in rat liver microsomes

目的考察英菲格拉替尼及其活性代谢产物BHS697、CQM157对大鼠肝微粒体中细胞色素P450(CYPs)和尿苷二磷酸葡萄糖醛酸转移酶(UGTs)的抑制作用。方法用体外孵育法,将待测化合物(英菲格拉替尼、BHS697或CQM157)、大鼠肝微粒体分别与CYP2B6、CYP2C8、CYP2C9、CYP2C19、CYP2D6、CYP3A4的特异性探针底物共孵育或分别与UGT1A1、UGT1A3、UGT1A6、UGT1A9、UGT2B7的特异性探针底物共孵育,用高效液相色谱-串联质谱法检测相应特征代谢物的生成量,用GraphPad Prism 8.0计算半数抑制浓度(IC_(50))和抑制常数(K_(i))。结果英菲格拉替尼、BHS697和CQM157对大鼠CYP2B6、CYP2C8、CYP2C9、CYP2C19、CYP2D6、CYP3A4和UGT1A6、UGT2B7的抑制较弱,IC_(50)值均大于10μmol·L^(-1);对UGT1A1具有中等抑制作用,IC_(50)值分别为2.70、3.17、7.43μmol·L^(-1)。英菲格拉替尼对大鼠UGT1A9和CQM157对大鼠UGT1A3均具有中等抑制作用,IC_(50)值分别为5.61、9.57μmol·L^(-1)。可逆性抑制分析发现,英菲格拉替尼、BHS697和CQM157均非竞争性抑制大鼠UGT1A1,K_(i)值分别为1.83、2.51、5.84μmol·L^(-1)。结论英菲格拉替尼对大鼠UGT1A1和UGT1A9具有中等抑制作用,其活性代谢产物BHS697、CQM157对大鼠UGT1A1也具有中等抑制作用。

Objective To evaluate the inhibitory effects of infigratinib and its pharmacologically active metabolites,BHS697 and CQM157,on cytochrome P450(CYPs)and UDP-glucuronosyltransferases(UGTs)in rat liver microsomes.Methods By adopting in vitro incubation method,the tested compounds(infigratinib,BHS697 or CQM157)and rat liver microsomes were incubated with the specific probe substrates of CYP2B6,CYP2C8,CYP2C9,CYP2C19,CYP2D6,CYP3A4,respectively,or the specific probe substrates of UGT1A1,UGT1A3,UGT1A6,UGT1A9,UGT2B7,respectively.The production of characteristic metabolites was detected by high performance liquid chromatographytandem mass spectrometry.The half maximal inhibitory concentration(IC_(50))and inhibition constant(K_(i))were calculated by Graph Pad Prism8.0.Results Infigratinib,BHS697 and CQM157 weakly inhibited CYP2 B6,CYP2 C8,CYP2 C9,CYP2 C19,CYP2 D6,CYP3 A4 and UGT1A6,UGT2B7 in rat liver microsomes,with IC_(50)values all more than 10μmol·L^(-1),and moderately inhibited UGT1A1 with IC_(50)values of 2.70,3.17,7.43μmol·L^(-1),respectively.Infigratinib had a moderate inhibitory effect on UGT1A9 and CQM157 had a moderate inhibitory effect on UGT1A3,with IC_(50)values of 5.61 and 9.57μmol·L^(-1),respectively.The reversible inhibition analysis showed that infigratinib,BHS697 and CQM157 all non-competitively inhibited UGT1A1,with K_(i)values of 1.83,2.51 and 5.84μmol·L^(-1),respectively.Conclusion Infigratinib had moderate inhibitory effects on UGT1A1 and UGT1A9 in rat liver microsomes and its pharmacologically active metabolites,BHS697 and CQM157,also had moderate inhibitory effects on UGT1A1.