Blanc Mettral Jaurès, Faller Nicolas, Cruchon Sandra, Sottas Loïc, Buclin Thierry, Schild Laurent, Choong Eva, Nahimana Aimable, Decosterd Laurent A
Laboratory of Clinical Pharmacology, Lausanne University Hospital, University of Lausanne, Lausanne, Switzerland.
Department of Pharmacology and Toxicology, Faculty of Biology and Medicine, Lausanne University Hospital, University of Lausanne, Lausanne, Switzerland.
Drug Metab Lett. 2019;13(2):102-110. doi: 10.2174/1872312813666190207150207.
Cancer cells undergo genetic and environmental changes that can alter cellular disposition of drugs, notably by alterations of transmembrane drug transporters expression. Whether the influx organic cation transporter 1 (OCT1) encoded by the gene SLC221A1 is implicated in the cellular uptake of imatinib is still controversial. Besides, imatinib ionization state may be modulated by the hypoxic acidic surrounding extracellular microenvironment.
To determine the functional contribution of OCTs and extracellular pH on imatinib cellular disposition.
We measured imatinib uptake in two different models of selective OCTs drug transporter expression (transfected Xenopus laevis oocytes and OCT-expressing HEK293 human cells), incubated at pH 7.4 and 6, using specific mass spectrometry analysis.
Imatinib cellular uptake occurred independently of OCT1- OCT2- or OCT3-mediated drug transport at pH 7.4. Uptake of the OCTs substrate tetraethylammonium in oocytes remained intact at pH 6, while the accumulation of imatinib in oocytes was 10-fold lower than at pH 7.4, irrespectively of OCTs expressions. In OCT1- and OCT2-HEK cells at pH 6, imatinib accumulation was reduced by 2- 3-fold regardless of OCTs expressions. Since 99.5% of imatinib at pH6 is under the cationic form, the reduced cellular accumulation of imatinib at such pH may be explained by the lower amount of uncharged imatinib remaining for passive diffusion across cellular membrane.
Imatinib is not a substrate of OCTs 1-3 while the environmental pH modulates cellular disposition of imatinib. The observation that a slightly acidic extracellular pH influences imatinib cellular accumulation is important, considering the low extracellular pH reported in the hematopoietic leukemia/ cancer cell microenvironment.
癌细胞会经历基因和环境变化,这些变化可改变药物的细胞分布,尤其是通过改变跨膜药物转运蛋白的表达。由基因SLC22A1编码的有机阳离子转运体1(OCT1)是否参与伊马替尼的细胞摄取仍存在争议。此外,伊马替尼的离子化状态可能会受到细胞外微环境缺氧酸性环境的调节。
确定OCTs和细胞外pH对伊马替尼细胞分布的功能贡献。
我们使用特定的质谱分析方法,在pH 7.4和pH 6条件下孵育,测量了两种不同的选择性OCTs药物转运蛋白表达模型(转染的非洲爪蟾卵母细胞和表达OCT的HEK293人细胞)中伊马替尼的摄取情况。
在pH 7.4时,伊马替尼的细胞摄取独立于OCT1、OCT2或OCT3介导的药物转运。在pH 6时,卵母细胞中OCTs底物四乙铵的摄取保持完整,而无论OCTs表达如何,伊马替尼在卵母细胞中的积累比在pH 7.4时低10倍。在pH 6的OCT1和OCT2-HEK细胞中,无论OCTs表达如何,伊马替尼的积累都减少了2至3倍。由于在pH 6时99.5%的伊马替尼处于阳离子形式,伊马替尼在这种pH下细胞积累减少可能是由于用于被动扩散穿过细胞膜的不带电荷的伊马替尼量较低。
伊马替尼不是OCTs 1-3的底物,而环境pH调节伊马替尼的细胞分布。考虑到造血白血病/癌细胞微环境中报道的低细胞外pH,细胞外pH略酸性影响伊马替尼细胞积累这一观察结果很重要。