ADME Sciences, Medicine Design, Worldwide Research and Development, Pfizer Inc., Groton, Connecticut (Y.-a.B., S.R., A.D.R., M.V.S.V.) and Modeling and Simulations Group, Medicine Design, Worldwide Research and Development, Pfizer Inc., Cambridge, Massachusetts (D.A.T.).
ADME Sciences, Medicine Design, Worldwide Research and Development, Pfizer Inc., Groton, Connecticut (Y.-a.B., S.R., A.D.R., M.V.S.V.) and Modeling and Simulations Group, Medicine Design, Worldwide Research and Development, Pfizer Inc., Cambridge, Massachusetts (D.A.T.)
Drug Metab Dispos. 2021 Jan;49(1):72-83. doi: 10.1124/dmd.120.000134. Epub 2020 Nov 2.
Current challenges with the in vitro-in vivo extrapolation (IVIVE) of hepatic uptake clearance involving organic anion-transporting polypeptide (OATP) 1B1/1B3 hinder drug design strategies. Here we evaluated the effect of 100% human plasma on the uptake clearance using transfected human embryonic kidney (HEK) 293 cells and primary human hepatocytes and assessed IVIVE. Apparent unbound uptake clearance (PS) increased significantly ( < 0.05) in the presence of plasma (vs. buffer incubations) for about 50% of compounds in both OATP1B1-transfected and wild-type HEK cells. Thus, plasma showed a minimal effect on the uptake ratios. With cultured human hepatocytes, plasma significantly ( < 0.05) increased PS for 11 of 19 OATP1B substrates (median change of 2.1-fold). Cell accumulation in HEK cells and hepatocytes was also increased for tolbutamide, which is not an OATP substrate. Plasma-to-buffer ratio of PS obtained in hepatocytes showed a good correlation with unbound fraction in plasma, and the relationship was best described by a "facilitated-dissociation" model. IVIVE was evaluated for the 19 OATP1B substrates using hepatocyte data in the presence of buffer and plasma. PS from buffer incubations markedly underpredicted hepatic intrinsic clearance (calculated via well stirred and parallel tube models) with an estimated bias of 0.10-0.13. Predictions improved when using PS from plasma incubations; however, considerable systemic underprediction was still apparent (0.19-0.26 bias). Plasma data with a global scaling factor of 3.8-5.3 showed good prediction accuracy (95% predictions within 3-fold; average fold error = 1.7, bias = 1). In summary, this study offers insight into the effect of plasma on the uptake clearance and its scope in improving IVIVE. SIGNIFICANCE STATEMENT: Our study using diverse anionic compounds shows that human plasma facilitates organic anion-transporting polypeptide 1B-mediated as well as passive uptake clearance, particularly for the highly bound compounds. Leveraging data from transfected human embryonic kidney 293 cells and primary human hepatocytes, we further evaluated mechanisms involved in the observed plasma-facilitated uptake transport. Enhanced hepatic uptake rate in the presence of plasma could be of relevance, as such mechanisms likely prevail in vivo and emphasize the need to maintain physiologically relevant assay conditions to achieve improved translation of transport data.
目前，涉及有机阴离子转运多肽（OATP）1B1/1B3 的肝摄取清除率的体外-体内外推（IVIVE）存在挑战，这阻碍了药物设计策略的发展。在这里，我们评估了使用转染的人胚肾（HEK）293 细胞和原代人肝细胞的 100%人血浆对摄取清除率的影响，并评估了 IVIVE。与缓冲液孵育相比，在 OATP1B1 转染和野生型 HEK 细胞中，约 50%的化合物在存在血浆时，表观未结合摄取清除率（PS）显著增加（<0.05）。因此，血浆对摄取比的影响很小。使用培养的人肝细胞时，19 种 OATP1B 底物中的 11 种（中位数变化 2.1 倍）的 PS 显著增加（<0.05）。甲苯磺丁脲在 HEK 细胞和肝细胞中的细胞积累也增加，而甲苯磺丁脲不是 OATP 底物。在肝细胞中获得的血浆与缓冲液的 PS 血浆与血浆中未结合分数呈良好相关性，且关系最好用“促进解离”模型描述。使用缓冲液孵育的 PS 对 19 种 OATP1B 底物的 IVIVE 进行了评估。来自缓冲液孵育的 PS 明显低估了肝内在清除率（通过搅拌良好和平行管模型计算），估计偏差为 0.10-0.13。当使用来自血浆孵育的 PS 时，预测得到改善；然而，仍然明显存在全身低估（0.19-0.26 偏差）。血浆数据与全球缩放因子 3.8-5.3 具有良好的预测准确性（95%预测值在 3 倍以内；平均倍误差=1.7，偏差=1）。总之，本研究深入了解了血浆对摄取清除率的影响及其在改善 IVIVE 中的作用。本研究使用不同的阴离子化合物表明，人血浆促进有机阴离子转运蛋白 1B 介导的以及被动摄取清除率，特别是对高度结合的化合物。利用转染的人胚肾 293 细胞和原代人肝细胞的数据，我们进一步评估了观察到的血浆促进摄取转运中涉及的机制。在血浆存在的情况下，增强的肝摄取率可能具有相关性，因为此类机制可能在体内普遍存在，并强调需要维持生理相关的测定条件以实现转运数据的更好转化。
