Meeting the Challenge of Predicting Hepatic Clearance of Compounds Slowly Metabolized by Cytochrome P450 Using a Novel Hepatocyte Model, HepatoPac
Meeting the Challenge of Predicting Hepatic Clearance of Compounds Slowly Metabolized by Cytochrome P450 Using a Novel Hepatocyte Model, HepatoPac
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DOI：
10.1124/dmd.113.053397
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发表时间：
2013-12-01
影响因子：
3.9
通讯作者：
Tweedie, Donald
中科院分区：
文献类型：
--
作者：
Chan, Tom S.;Yu, Hongbin;Tweedie, Donald
生成准确的体外固有清除率数据是预测体内人体清除率的一个重要方面。悬浮液中的原代肝细胞通常用于预测体内清除率;然而，孵育时间通常限于4-6小时，这不足以准确评价缓慢代谢化合物的代谢稳定性。HepatoPac是一种微模式化的肝细胞-成纤维细胞共培养系统，可用于连续孵育长达7天。本研究评估了人HepatoPac预测17种市售化合物体内清除率（CL）的能力，这些化合物具有低至中等清除率（< 12 ml/min/kg）。使用充分搅拌模型将各化合物的体外消失半衰期转换为肝脏清除率，校正和不校正血浆蛋白结合。使用3个个体供体，准确预测了17种化合物中10种的肝脏CL（59%;预测清除率在观察到的人体体内清除率值的2倍范围内）。预测准确度增加至76%（17种化合物中的13种），验收标准定义为3倍以内。当仅考虑代表17种化合物中的10种的低清除率化合物（< 5 ml/min/kg）时，预测的准确度在2倍内为60%，在3倍内为90%。此外，将HepatoPac中三种缓慢代谢化合物（阿普唑仑、美洛昔康和甲苯磺丁脲）的周转率与悬浮肝细胞中的周转率直接进行比较。与悬浮肝细胞相比，使用HepatoPac时阿普唑仑和甲苯磺丁脲的转化率约为2倍，这与外推值大致一致（校正HepatoPac的孵育时间较长和细胞数量较低）。HepatoPac（而非悬浮肝细胞）显示出美洛昔康的显著转换。这些结果证明了HepatoPac用于预测体内肝脏清除率的实用性，特别是对于低清除率化合物。
Generating accurate in vitro intrinsic clearance data is an important aspect of predicting in vivo human clearance. Primary hepatocytes in suspension are routinely used to predict in vivo clearance; however, incubation times have typically been limited to 4-6 hours, which is not long enough to accurately evaluate the metabolic stability of slowly metabolized compounds. HepatoPac is a micropatterened hepatocyte-fibroblast coculture system that can be used for continuous incubations of up to 7 days. This study evaluated the ability of human HepatoPac to predict the in vivo clearance (CL) of 17 commercially available compounds with low to intermediate clearance (< 12 ml/min per kg). In vitro half-life for disappearance of each compound was converted to hepatic clearance using the well stirred model, with and without correction for plasma protein binding. Hepatic CL, using three individual donors, was accurately predicted for 10 of 17 compounds (59%; predicted clearance within 2-fold of observed human in vivo clearance values). The accuracy of prediction increased to 76% (13 of 17 compounds) with an acceptance criterion defined as within 3-fold. When considering only low clearance compounds (< 5 ml/min per kg), which represented 10 of the 17 compounds, the accuracy of prediction was 60% within 2-fold and 90% within 3-fold. In addition, the turnover of three slowly metabolized compounds (alprazolam, meloxicam, and tolbutamide) in HepatoPac was directly compared with turnover in suspended hepatocytes. The turnover of alprazolam and tolbutamide was approximately 2-fold greater using HepatoPac compared with suspended hepatocytes, which was roughly in line with the extrapolated values (correcting for the longer incubation time and lower cell number with HepatoPac). HepatoPac, but not suspended hepatocytes, demonstrated significant turnover of meloxicam. These results demonstrate the utility of HepatoPac for prediction of in vivo hepatic clearance, particularly with low clearance compounds.