Inhibition and stimulation of the human breast cancer resistance protein as in vitro predictor of drug–drug interactions of drugs of abuse
Inhibition and stimulation of the human breast cancer resistance protein as in vitro predictor of drug–drug interactions of drugs of abuse
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DOI：
10.1007/s00204-018-2276-y
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发表时间：
2018-08
影响因子：
6.1
通讯作者：
Lea Wagmann;H. Maurer;M. Meyer
中科院分区：
文献类型：
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作者：
Lea Wagmann;H. Maurer;M. Meyer
转运蛋白介导的药物-药物相互作用(DDI)可能导致不良临床事件。由于滥用药物(DOA)在市场上没有临床前的安全性研究，关于与膜转运蛋白相互作用的信息非常有限。因此，对13个不同类别的DOA进行了体外亲和力测试，以检测它们与人乳腺癌耐药蛋白(HBCRP)的亲和力，hBCRP是一种重要的外排转运蛋白。由于腺苷5‘-三磷酸(ATP)的水解对hBCRP活性至关重要，因此测定了腺苷5’-二磷酸(ADP)的形成，并将其作为hBCRP ATPase活性的体外标志物。采用亲水作用高效液相色谱与高分辨串联质谱仪联用进行ADP定量，并与hBCRP底物柳氮磺吡啶或hBCRP抑制剂正钒酸作参比孵育时的ADP含量进行比较。如果DOA引起刺激或抑制，则进行进一步的研究，如Michaelis-Menten动力学模型或IC50值测定。在所测试的DOA中，有7种化合物对hBCRP-ATPase有显著的刺激作用。内分泌激素3，4-BDB和植物生物碱美托宁的刺激性最强。它们对hBCRP ATPase的亲和力低于柳氮磺胺吡啶，但与另一种hBCRP模型底物瑞舒伐他汀相当。5种DOA对hBCRP-ATPase的抑制有统计学意义。IC50值的测定表明，合成的大麻素受体激动剂JWH-200和Win 55,212-2是与原钒酸相当的最强的抑制剂。目前的研究清楚地表明，在模型底物或抑制剂的范围内，测试的DOA部分显示出对hBCRP的高亲和力。因此，存在hBCRP介导的DDI的风险，这需要在临床环境中考虑。
Transporter-mediated drug–drug interactions (DDI) may induce adverse clinical events. As drugs of abuse (DOA) are marketed without preclinical safety studies, only very limited information about interplay with membrane transporters are available. Therefore, 13 DOA of various classes were tested for their in vitro affinity to the human breast cancer resistance protein (hBCRP), an important efflux transporter. As adenosine 5′-triphosphate (ATP) hydrolysis is crucial for hBCRP activity, adenosine 5′-diphosphate (ADP) formation was measured and used as in vitro marker for hBCRP ATPase activity. ADP quantification was performed by hydrophilic interaction liquid chromatography coupled to high-resolution tandem mass spectrometry and its amount in test compound incubations was compared to that in reference incubations using the hBCRP substrate sulfasalazine or the hBCRP inhibitor orthovanadate. If DOA caused stimulation or inhibition, further investigations such as Michaelis–Menten kinetic modeling or IC50value determination were conducted. Among the tested DOA, seven compounds showed statistically significant hBCRP ATPase stimulation. The entactogen 3,4-BDB and the plant alkaloid mitragynine were identified as strongest stimulators. Their affinity to the hBCRP ATPase was lower than that of sulfasalazine but comparable to that of rosuvastatin, another hBCRP model substrate. Five DOA showed statistically significant hBCRP ATPase inhibition. Determination of IC50values identified the synthetic cannabinoid receptor agonists JWH-200 and WIN 55,212-2 as the strongest inhibitors comparable to orthovanadate. The present study clearly demonstrated that tested DOA show in part high affinities to the hBCRP within the range of model substrates or inhibitors. Thus, there is a risk of hBCRP-mediated DDI, which needs to be considered in clinical settings.