Characterization of the transport of nucleoside analog drugs by the human multidrug resistance proteins MRP4 and MRP5
Characterization of the transport of nucleoside analog drugs by the human multidrug resistance proteins MRP4 and MRP5
复制标题
DOI：
10.1124/mol.63.5.1094
复制
发表时间：
2003-05-01
影响因子：
3.6
通讯作者：
Borst, P
中科院分区：
文献类型：
--
作者：
Reid, G;Wielinga, P;Borst, P
人多药耐药蛋白MRP 4和MRP 5是有机阴离子转运蛋白，具有转运环核苷酸和一些核苷单磷酸类似物的能力。在癌症和病毒感染的化疗中使用的碱基和核苷类似物是潜在的底物。为了评估MRP 4和MRP 5对这些药物耐药性的可能贡献，我们研究了MRP 4和MRP 5介导的转运。在细胞毒性试验中，MRP 4赋予抗病毒剂9-（2-膦酰甲氧基乙基）腺嘌呤（PMEA）的抗性，高效液相色谱分析表明，与MRP 5一样，MRP 4以未修饰的形式转运PMEA。MRP 4还介导了对其他非环核苷膦酸酯的实质性耐药性，而MRP 5则没有。除了低水平的MRP 4介导的克拉屈滨耐药性外，临床使用的抗癌核苷的细胞毒性不受MRP 4或MRP 5过表达的影响。相比之下，MRP 5介导了基于嘧啶的抗病毒2 '，3'-双脱氧核苷2 '，3'-二氨基-2 '，3'-双脱氧胸苷5 '-单磷酸（d4 TMP）及其氨基磷酸酯衍生物丙氨酰-d4 TMP从分别负载有2'，3 '-二氨基-2'，3 '-双脱氧胸苷前药环水杨酰-d4 TMP和芳氧基氨基磷酸酯d4 TMP（So 324）的细胞中的流出。此外，只有来自MRP 5过表达细胞的内翻膜囊泡积累丙氨酸-d4 TMP。进行了细胞外排和囊泡摄取研究，以进一步比较MRP 4和MRP 5介导的转运，结果表明双嘧达莫、地拉卓、硝基苯甲基巯基嘌呤核苷、西地那非、曲喹辛和MK 571对MRP 4的抑制作用大于MRP 5，而环核苷酸和单磷酸化核苷类似物对这两种泵的抑制作用同样较差。这些结果强烈表明，MRP 4和MRP 5对基于核苷酸的底物的亲和力较低。
The human multidrug resistance proteins MRP4 and MRP5 are organic anion transporters that have the unusual ability to transport cyclic nucleotides and some nucleoside monophosphate analogs. Base and nucleoside analogs used in the chemotherapy of cancer and viral infections are potential substrates. To assess the possible contribution of MRP4 and MRP5 to resistance against these drugs, we have investigated the transport mediated by MRP4 and MRP5. In cytotoxicity assays, MRP4 conferred resistance to the antiviral agent 9-(2-phosphonomethoxyethyl) adenine (PMEA) and high-performance liquid chromatography analysis showed that, like MRP5, MRP4 transported PMEA in an unmodified form. MRP4 also mediated substantial resistance against other acyclic nucleoside phosphonates, whereas MRP5 did not. Apart from low-level MRP4-mediated cladribine resistance, the cytotoxicity of clinically used anticancer nucleosides was not influenced by overexpression of MRP4 or MRP5. In contrast, MRP5 mediated efflux of the pyrimidine-based antiviral 2',3'-dideoxynucleoside 2',3 '-didehydro-2',3'-dideoxythymidine 5'-monophosphate (d4TMP) and its phosphoramidate derivative alaninyl-d4TMP from cells loaded with the 2',3'-didehydro-2',3'-dideoxythymidine prodrugs cyclosaligenyl-d4TMP and aryloxyphosphoramidate d4TMP (So324), respectively. Moreover, only inside-out membrane vesicles derived from MRP5-overexpressing cells accumulated alaninyl-d4TMP. Cellular efflux and vesicular uptake studies were carried out to further compare transport mediated by MRP4 and MRP5 and showed that dipyridamole, dilazep, nitrobenzyl mercaptopurine riboside, sildenafil, trequinsin and MK571 inhibited MRP4 more than MRP5, whereas cyclic nucleotides and monophosphorylated nucleoside analogs were equally poor inhibitors of both pumps. These results strongly suggest that the affinity of MRP4 and MRP5 for nucleotide-based substrates is low.