Byer-Alcorace Alexander, Thomas Cody, Taub Mitchell E, Piekos Stephanie
Department of Drug Metabolism and Pharmacokinetics, Boehringer Ingelheim Pharmaceuticals Inc, Ridgefield, Connecticut.
Department of Drug Metabolism and Pharmacokinetics, Boehringer Ingelheim Pharmaceuticals Inc, Ridgefield, Connecticut.
Drug Metab Dispos. 2025 Apr;53(4):100051. doi: 10.1016/j.dmd.2025.100051. Epub 2025 Feb 12.
Over the last several decades, efforts in medicinal chemistry have aimed to reduce the extent of CYP metabolism of new chemical entities. This approach, however, has led to increased susceptibility to metabolism by non-CYP-mediated pathways, particularly involving other phase I enzymes such as aldehyde oxidase (AO). Commonly used in vitro models, such as suspended or cocultured primary human hepatocytes, have limitations in evaluating the disposition of compounds metabolized by AO due to low or variable levels of enzyme activity. Thus, an in vitro model that exhibits high to moderate levels of AO activity that can better predict the contribution of AO to drug metabolism and its impact on drug clearance is needed. A novel, 2D+ primary human hepatocyte model, TruVivo, was evaluated for its potential utility to improve hepatic clearance (CL) predictions and determine the contribution of AO to drug metabolism in humans. TruVivo demonstrated stable levels of AO activity for at least 2 weeks that were higher than levels in other hepatocyte models. CL predictions generated using TruVivo for the reference compounds carbazeran, zoniporide, zaleplon, and O6-benzylguanine were within 2-fold of reported in vivo CL values. Furthermore, the estimated fraction metabolized by AO for zaleplon and zoniporide was within 25% of reported in vivo values, whereas that for carbazeran and O6-benzylguanine was similar to those generated in other systems. These findings suggest TruVivo may offer a novel means to assess CL of AO substrates more accurately, even over extended incubation times for low clearance compounds. SIGNIFICANCE STATEMENT: The TruVivo in vitro primary human hepatocyte model maintains high levels of aldehyde oxidase (AO) activity for at least 14 days in culture, making the system suitable for evaluating slowly metabolized compounds, particularly those metabolized by AO. This novel system may therefore be useful for improving human clearance predictions for AO substrates.
在过去几十年中，药物化学领域一直致力于降低新化学实体的细胞色素P450（CYP）代谢程度。然而，这种方法导致了新化学实体对非CYP介导途径代谢的易感性增加，特别是涉及其他I相酶，如醛氧化酶（AO）。常用的体外模型，如悬浮或共培养的原代人肝细胞，由于酶活性低或变化大，在评估AO代谢化合物的处置方面存在局限性。因此，需要一种体外模型，该模型具有高至中等水平的AO活性，能够更好地预测AO对药物代谢的贡献及其对药物清除率的影响。一种新型的二维+原代人肝细胞模型TruVivo，评估了其在改善肝清除率（CL）预测以及确定AO对人体药物代谢贡献方面的潜在效用。TruVivo在至少2周内表现出稳定的AO活性水平，高于其他肝细胞模型中的水平。使用TruVivo对参考化合物卡巴西兰、佐尼普明、扎来普隆和O6-苄基鸟嘌呤进行的CL预测在报告的体内CL值的2倍以内。此外，扎来普隆和佐尼普明经AO代谢的估计分数在报告的体内值的25%以内，而卡巴西兰和O6-苄基鸟嘌呤的该分数与其他系统中的结果相似。这些发现表明，即使对于低清除率化合物延长孵育时间，TruVivo也可能提供一种更准确评估AO底物CL的新方法。重要声明：TruVivo体外原代人肝细胞模型在培养中至少14天保持高水平的醛氧化酶（AO）活性，使该系统适用于评估缓慢代谢的化合物，特别是那些由AO代谢的化合物。因此，这个新系统可能有助于改善对AO底物的人体清除率预测。
