Studies comparing in vivo:in vitro metabolism of three pharmaceutical compounds in rat, dog, monkey, and human using cryopreserved hepatocytes, microsomes, and collagen gel immobilized hepatocyte cultures.
Studies comparing in vivo:in vitro metabolism of three pharmaceutical compounds in rat, dog, monkey, and human using cryopreserved hepatocytes, microsomes, and collagen gel immobilized hepatocyte cultures.
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发表时间：
2001-07
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通讯作者：
N. Hewitt;Karl-Uhlrich Bühring;J. Dasenbrock;J. Haunschild;B. Ladstetter;D. Utesch
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作者：
N. Hewitt;Karl-Uhlrich Bühring;J. Dasenbrock;J. Haunschild;B. Ladstetter;D. Utesch
比较了三种药物化合物 EMD68843、EMD96785 和 EMD128130 在新鲜和冷冻保存的肝细胞 (CPH) 悬浮液和大鼠、狗、猴子和人肝脏微粒体以及新鲜人和大鼠肝细胞胶原凝胶固定培养物 (GIC) 中的体内代谢。一半的主要体内代谢物是由第一阶段（羟基化、氧化、水解、N-脱烷基化）产生的，一半是由第二阶段代谢（主要是葡萄糖醛酸化，但也有硫酸化和甘氨酸缀合）产生的。肝细胞中产生的每种化合物的 1 相和 2 相代谢物的特性和百分比与每个物种体内的代谢物的特性和百分比进行了很好的比较。 GIC 中的葡萄糖醛酸化作用比 CPH 中更广泛。相比之下，CPH 而非 GIC 产生硫酸盐代谢物。微粒体（仅补充 NADPH）产生大部分 1 相代谢物，但不产生 2 相代谢物。 CPH 中的代谢与新鲜肝细胞悬浮液中的代谢相同。通过 CPH 和微粒体检测代谢中离散物种的差异。 CPH 的细胞色素 P450 和葡萄糖醛酸基 S-转移酶含量不能解释这些细胞中 1 相和 2 相代谢物百分比或母体化合物消失率的物种差异。这些数据显示体内和体外形成的主要代谢物之间具有良好的相关性。与微粒体不同，CPH 和 GIC 进行连续的 1 期和 2 期代谢。每个体外系统都有其自身的优势，然而，对于短期代谢研究，CPH 可能更有用，因为它们比 GIC 更容易获得、更容易和更快地制备，并且比微粒体具有更全面的酶系统。
The in vivo metabolism of three pharmaceutical compounds, EMD68843, EMD96785, and EMD128130, was compared in fresh and cryopreserved hepatocyte (CPH) suspensions and microsomes from rat, dog, monkey, and human livers and fresh human and rat hepatocyte collagen gel immobilized cultures (GICs). Half of the major in vivo metabolites was produced by phase 1 (hydroxylation, oxidation, hydrolysis, N-dealkylation) and half by phase 2 metabolism (mostly glucuronidation but also sulfation and glycine conjugation). The identity and percentage of phase 1 and 2 metabolites from each compound produced in hepatocytes compared well with that in each species in vivo. Glucuronidation was more extensive in GICs than in CPHs. In contrast, CPHs but not GICs, produced sulfate metabolites. Microsomes (supplemented with NADPH only) produced most of the phase 1 but no phase 2 metabolites. Metabolism in CPHs was the same as in fresh hepatocyte suspensions. Discrete species differences in metabolism were detected by CPHs and microsomes. Cytochrome P450 and glucuronosyl S-transferase contents of CPHs did not account for species differences in the percentage of phase 1 and 2 metabolites or the rate of disappearance of the parent compounds in these cells. These data show a good correlation between major metabolites formed in vivo and in vitro. CPHs and GICs, unlike microsomes, carried out sequential phase 1 and 2 metabolism. Each in vitro system has its own advantages, however, for short-term metabolism studies CPHs may be more useful since they are readily available, easier and quicker to prepare than GICs, and have more comprehensive enzyme systems than microsomes.