Doricakova Aneta, Theile Dirk, Weiss Johanna, Vrzal Radim
Department of Cell Biology and Genetics, Faculty of Science, Palacky University, Slechtitelu 27, 783 71, Olomouc, Czech Republic.
Department of Clinical Pharmacology and Pharmacoepidemiology, University of Heidelberg, Im Neuenheimer Feld 410, 69120, Heidelberg, Germany.
Naunyn Schmiedebergs Arch Pharmacol. 2017 Jan;390(1):49-59. doi: 10.1007/s00210-016-1304-9. Epub 2016 Sep 27.
The pregnane X receptor (PXR) is a transcription factor regulating P-glycoprotein (P-gp; ABCB1)-mediated transport and cytochrome P450 3A4 (CYP3A4)-mediated metabolism of xenobiotics thereby affecting the pharmacokinetics of many drugs and potentially modulating clinical efficacy. Thus, pharmacokinetic drug-drug interactions can arise from PXR activation. Here, we examined whether the selective α1-adrenoreceptor blocker tamsulosin or the antagonist of muscarinic receptors tolterodine affect PXR-mediated regulation of CYP3A4 and of P-gp at the messenger RNA (mRNA) and protein level in an enantiomer-specific way. In addition, the effect of tamsulosin and tolterodine on P-gp activity was evaluated. We used quantitative real-time PCR, gene reporter assay, western blotting, rhodamine efflux assay, and calcein assay for determination of expression, activity, and inhibition of P-glycoprotein. The studied compounds significantly and concentration-dependently increased PXR activity in the ABCB1-driven luciferase-based reporter gene assay. We observed much stronger induction of ABCB1 mRNA by S-tamsulosin as compared to the R or racemic form. R or racemic form of tolterodine and R-tamsulosin concentration-dependently increased P-gp protein expression; the latter also enhanced P-gp efflux function in a rhodamine-based efflux assay. R-tamsulosin and all forms of tolderodine slightly inhibited P-gp. The effect on CYP3A4 expression followed the same pattern but was much weaker. Taken together, tamsulosin and tolterodine are demonstrated to interfere with P-gp and CYP3A4 regulation in an enantiomer-specific way.
孕烷X受体(PXR)是一种转录因子,可调节P-糖蛋白(P-gp;ABCB1)介导的外源性物质转运以及细胞色素P450 3A4(CYP3A4)介导的外源性物质代谢,从而影响许多药物的药代动力学,并可能调节临床疗效。因此,PXR激活可引发药代动力学方面的药物相互作用。在此,我们研究了选择性α1-肾上腺素能受体阻滞剂坦索罗辛或毒蕈碱受体拮抗剂托特罗定是否以对映体特异性方式在信使核糖核酸(mRNA)和蛋白质水平上影响PXR介导的CYP3A4和P-gp的调节。此外,还评估了坦索罗辛和托特罗定对P-gp活性的影响。我们使用定量实时聚合酶链反应、基因报告基因检测、蛋白质免疫印迹、罗丹明外排检测和钙黄绿素检测来测定P-糖蛋白的表达、活性和抑制情况。在基于ABCB1驱动的荧光素酶报告基因检测中,所研究化合物显著且呈浓度依赖性地增加了PXR活性。我们观察到,与R型或消旋体形式相比,S-坦索罗辛对ABCB1 mRNA的诱导作用更强。R型或消旋体形式的托特罗定以及R-坦索罗辛呈浓度依赖性地增加P-gp蛋白表达;后者在基于罗丹明的外排检测中还增强了P-gp的外排功能。R-坦索罗辛和所有形式的托特罗定都对P-gp有轻微抑制作用。对CYP3A4表达的影响遵循相同模式,但作用较弱。综上所述,已证明坦索罗辛和托特罗定以对映体特异性方式干扰P-gp和CYP3A4的调节。