Metabolism of propafenone and verapamil by cryopreserved human, rat, mouse and dog hepatocytes: comparison with metabolism in vivo
Metabolism of propafenone and verapamil by cryopreserved human, rat, mouse and dog hepatocytes: comparison with metabolism in vivo
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DOI：
10.1007/s00210-004-0875-z
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发表时间：
2004-04-01
影响因子：
3.6
通讯作者：
Hengstler, JG
中科院分区：
文献类型：
--
作者：
Reder-Hilz, B;Ullrich, M;Hengstler, JG
在本研究中，我们使用冻存的人、犬（比格犬）、大鼠（Sprague-Dawley）和小鼠（NMRI）肝细胞检查了[C-14]普罗帕酮（P）和[C-14]维拉帕米（V）的代谢。提取后，通过HPLC结合UV和放射性检测鉴别代谢产物的百分比。使用β-葡萄糖醛酸酶裂解II相代谢物。将冻存肝细胞的药物代谢与体内相应物种的代谢进行比较，在与冻存肝细胞孵育后，鉴定了体内实验中已知的所有I相和II相代谢产物：5-羟基-P（5-OH-P）、4 '-羟基-P（4'-OH-P）、N-去丙基-P（NdesP）和相应的葡萄糖醛酸苷。普罗帕酮羟基化的优先位置存在种间差异：与冻存的人（n=4）、犬（n= 3）、大鼠（n = 3）和小鼠（n=4）肝细胞孵育后，5-OH-P分别占总代谢产物的91%、51%、16%和3%。这些结果与体内实验中已知的种间差异一致。V的代谢比P的代谢更复杂。然而，在与所有四个种属的冻存肝细胞孵育后，鉴定了体内实验中已知的所有I相代谢物和预期的葡萄糖醛酸苷。正如预期的结果，在体内实验中，有没有重大的种间差异，相对于第一阶段的代谢产物，虽然维拉帕米第一阶段的代谢产物的冻存犬肝细胞的共轭是远远弱于其他species.In结论，第一阶段和第二阶段的代谢P和V进行了评价，使用肝细胞在体外。在与冻存肝细胞悬液短期孵育后，鉴定了体内实验中已知的所有相关种属间差异。
In the present study we examined the metabolism of [C-14]propafenone (P) and [C-14]verapamil (V) using cryopreserved human, dog (Beagle), rat (Sprague-Dawley) and mouse (NMRI) hepatocytes. The percentage ratios of the metabolites were identified after extraction by HPLC with UV and radioactivity detection. Phase-II metabolites were cleaved using beta-glucuronidase. Metabolism of the drugs by cryopreserved hepatocytes was compared with that in the respective species in vivo.All phase-I and -II metabolites known from in vivo experiments: 5-hydroxy-P (5-OH-P); 4'-hydroxy-P (4'-OH-P); N-despropyl-P (NdesP) and the respective glucuronides, were identified after incubation with cryopreserved hepatocytes. Interspecies differences were observed concerning the preferential position of propafenone hydroxylation: 5-OH-P made up 91, 51, 16 and 3% of the total metabolites after incubation with cryopreserved human (n=4), dog (n=3), rat (n=3) and mouse (n=4) hepatocytes respectively. These results are consistent with interspecies differences known from in vivo experiments. The metabolism of V is more complex than that of P. Nevertheless, all phase-I metabolites known from in vivo experiments and the expected glucuronides were identified after incubation with cryopreserved hepatocytes from all four species. As expected from the results of in vivo experiments, there were no major interspecies differences with respect to phase-I metabolites although the conjugation of verapamil phase-I metabolites by cryopreserved canine hepatocytes was much weaker than for the other species.In conclusion, phase-I and phase-II metabolism of P and V was evaluated using hepatocytes in vitro. All of the relevant interspecies differences known from in vivo experiments were identified after short-term incubation with cryopreserved hepatocytes in suspension.