Medicine Design, Pfizer Inc., Cambridge, Massachusetts (J.E.M., A.S.K., Y.W.), and Medicine Design, Pfizer Inc., Groton, Connecticut (K.N., N.J., Z.L., T.C.G.).
Medicine Design, Pfizer Inc., Cambridge, Massachusetts (J.E.M., A.S.K., Y.W.), and Medicine Design, Pfizer Inc., Groton, Connecticut (K.N., N.J., Z.L., T.C.G.)
J Pharmacol Exp Ther. 2018 May;365(2):262-271. doi: 10.1124/jpet.117.247296. Epub 2018 Feb 12.
The potential for drug-drug interactions (DDIs) arising from transcriptional regulation of drug-disposition genes via activation of nuclear receptors (NRs), such as pregnane X receptor (PXR), constitutive androstane receptor (CAR), and aryl hydrocarbon receptor (AhR), remains largely unexplored, as highlighted in a recent guidance document from the European Medicines Agency. The goal of this research was to establish PXR-/CAR-/AhR-specific drug-metabolizing enzyme (DME) and transporter gene expression signatures in sandwich-cultured cryopreserved human hepatocytes using selective activators of PXR (rifampin), CAR (CITCO), and AhR (omeprazole). Dose response for ligand-induced changes to 38 major human DMEs and critical hepatobiliary transporters were assessed using a custom gene expression array card. We identified novel differentially expressed drug-disposition genes for PXR (↑ABCB1/MDR1, CYP2C9, CYP2C19, and EPHX1, ↓ABCB11), CAR [↑sulfotransferase (SULT) 1E1, uridine glucuronosyl transferase (UGT) 2B4], and AhR (↑SLC10A1/NTCP, SLCO1B1/OATP1B1], and coregulated genes (CYP1A1, CYP2B6, CYP2C8, CYP3A4, UGT1A1, UGT1A4). Subsequently, DME gene expression signatures were generated for known CYP3A4 inducers PF-06282999 and pazopanib. The former produced an induction signature almost identical to that of rifampin, suggesting activation of the PXR pathway, whereas the latter produced an expression signature distinct from those of PXR, CAR, or AhR, suggesting involvement of an alternate pathway(s). These results demonstrate that involvement of PXR/CAR/AhR can be identified via expression changes of signature DME/transporter genes. Inclusion of such signature genes could serve to simultaneously identify potential inducers and inhibitors, and the NRs involved in the transcriptional regulation, thus providing a more holistic and mechanism-based assessment of DDI risk for DMEs and transporters beyond conventional cytochrome P450 isoforms.
药物-药物相互作用（DDI）的潜力源于核受体（NRs）对药物处置基因的转录调控，例如孕烷 X 受体（PXR）、组成型雄烷受体（CAR）和芳香烃受体（AhR），这在欧洲药品管理局最近的一份指导文件中得到了强调。本研究的目的是在使用 PXR（利福平）、CAR（CITCO）和 AhR（奥美拉唑）的选择性激活剂的三明治培养冷冻保存人肝细胞中建立 PXR-/CAR-/AhR 特异性药物代谢酶（DME）和转运体基因表达特征。使用定制的基因表达阵列卡评估配体诱导的 38 种主要人类 DME 和关键肝胆转运体变化的剂量反应。我们确定了 PXR（↑ABCB1/MDR1、CYP2C9、CYP2C19 和 EPHX1、↓ABCB11）、CAR [↑磺基转移酶（SULT）1E1、尿苷葡萄糖醛酸基转移酶（UGT）2B4] 和 AhR（↑SLC10A1/NTCP、SLCO1B1/OATP1B1] 的新型差异表达药物处置基因，以及共调节基因（CYP1A1、CYP2B6、CYP2C8、CYP3A4、UGT1A1、UGT1A4）。随后，为已知的 CYP3A4 诱导剂 PF-06282999 和帕唑帕尼生成了 DME 基因表达特征。前者产生的诱导特征几乎与利福平相同，表明 PXR 途径的激活，而后者产生的表达特征与 PXR、CAR 或 AhR 不同，表明涉及替代途径（多个）。这些结果表明，通过特征性 DME/转运体基因的表达变化可以识别 PXR/CAR/AhR 的参与。包含此类特征基因可以同时识别潜在的诱导剂和抑制剂以及参与转录调控的 NRs，从而提供对 DME 和转运体的 DDI 风险的更全面和基于机制的评估，超越传统的细胞色素 P450 同工酶。
