Modeling Therapeutic Antibody-Small Molecule Drug-Drug Interactions Using a Three-Dimensional Perfusable Human Liver Coculture Platform
Modeling Therapeutic Antibody-Small Molecule Drug-Drug Interactions Using a Three-Dimensional Perfusable Human Liver Coculture Platform
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DOI：
10.1124/dmd.116.071456
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发表时间：
2016-12-01
影响因子：
3.9
通讯作者：
Griffith, Linda G.
中科院分区：
文献类型：
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作者：
Long, Thomas J.;Cosgrove, Patrick A.;Griffith, Linda G.
传统的体外人肝细胞培养模型在短期培养期间失去了关键的肝功能，例如代谢活性。先进的三维（3D）肝脏共培养平台提供了扩展肝细胞功能的潜力，并允许研究更复杂的生物相互作用，这可以改善和完善人类药物安全性评价。在这里，我们使用灌注流三维微反应器平台的冻存原代人肝细胞和枯否细胞的共培养研究细胞色素P450 3A 4亚型（CYP 3A 4）活性的调节慢性白细胞介素6（IL-6）介导的炎症超过2周。肝细胞培养物在2周内保持稳定，具有一致的白蛋白产生和基础IL-6水平。模拟炎症状态的直接IL-6刺激诱导CYP 3A 4活性的剂量依赖性抑制、C反应蛋白（CRP）分泌增加和脱落可溶性白细胞介素-6受体（IL-6 R）水平降低，表明预期的肝脏IL-6生物活性。托珠单抗是一种用于治疗类风湿性关节炎的抗IL-6 R单克隆抗体，临床上已证明其可通过调节细胞色素P450酶活性影响小分子药物的药代动力学，这一作用在传统的肝培养物中未观察到。我们现在已经概括了在3D生物反应器系统中的临床观察。在持续存在IL-6的情况下，托珠单抗显示出去抑制CYP 3A 4活性，同时在72小时后降低CRP浓度。CYP 3A 4活性的这种变化降低了小分子CYP 3A 4底物辛伐他汀羟基酸的半衰期和曲线下面积，直至末次可测量浓度（AUC（last）），在托珠单抗治疗前后测量。我们的结论是，下一代体外肝培养平台非常适合这些类型的长期治疗研究，并显示出改善药物安全性评估的前景。
Traditional in vitro human liver cell culture models lose key hepatic functions such as metabolic activity during short-term culture. Advanced three-dimensional (3D) liver coculture platforms offer the potential for extended hepatocyte functionality and allow for the study of more complex biologic interactions, which can improve and refine human drug safety evaluations. Here, we use a perfusion flow 3D microreactor platform for the coculture of cryopreserved primary human hepatocytes and Kupffer cells to study the regulation of cytochrome P450 3A4 isoform (CYP3A4) activity by chronic interleukin 6 (IL-6)-mediated inflammation over 2 weeks. Hepatocyte cultures remained stable over 2 weeks, with consistent albumin production and basal IL-6 levels. Direct IL-6 stimulation that mimics an inflammatory state induced a dose-dependent suppression of CYP3A4 activity, an increase in C-reactive protein (CRP) secretion, and a decrease in shed soluble interleukin-6 receptor (IL-6R) levels, indicating expected hepatic IL-6 bioactivity. Tocilizumab, an anti-IL6R monoclonal antibody used to treat rheumatoid arthritis, has been demonstrated clinically to impact small molecule drug pharmacokinetics by modulating cytochrome P450 enzyme activities, an effect not observed in traditional hepatic cultures. We have now recapitulated the clinical observation in a 3D bioreactor system. Tocilizumab was shown to desuppress CYP3A4 activity while reducing the CRP concentration after 72 hours in the continued presence of IL-6. This change in CYP3A4 activity decreased the half-life and area under the curve up to the last measurable concentration (AUC(last)) of the small molecule CYP3A4 substrate simvastatin hydroxy acid, measured before and after tocilizumab treatment. We conclude that next-generation in vitro liver culture platforms are well suited for these types of long-term treatment studies and show promise for improved drug safety assessment.