Cellular and biochemical characterization of VX-710 as a chemosensitizer: Reversal of P-glycoprotein-mediated multidrug resistance in vitro
Cellular and biochemical characterization of VX-710 as a chemosensitizer: Reversal of P-glycoprotein-mediated multidrug resistance in vitro
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DOI：
10.1097/00001813-199702000-00004
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发表时间：
1997-02-01
影响因子：
2.3
通讯作者：
Harding, MW
中科院分区：
文献类型：
--
作者：
Germann, UA;Shlyakhter, D;Harding, MW
评价了VX-710或（S）-N-[2-氧代-2-（3，4，5-三甲氧基苯基）十六烷基]-哌啶-2-羧酸1，7-双（3-吡啶基）-4-庚酯（FK 506结合蛋白FKBP 12的新型非大环配体）逆转P-糖蛋白介导的多药耐药的体外能力。VX-710在0.5-5 μ M恢复多种多药耐药细胞对阿霉素，长春新碱，依托泊苷或紫杉醇的细胞毒性作用的敏感性，包括药物选择的人骨髓瘤和上皮癌细胞，以及人MDR 1 cDNA转染的小鼠白血病和成纤维细胞。摄取实验表明，VX-710在0.5-2.5 μ M完全恢复[C-14]阿霉素在多药耐药细胞的细胞内积累，这表明VX-710抑制P-糖蛋白的药物外排活性。VX-710可有效抑制[H-3]叠氮平或[I-125]碘芳基叠氮哌唑嗪对P-糖蛋白的光亲和标记，EC（50）值为0.75和0.55 μ M。此外，P-糖蛋白被VX-710的氚化光亲和类似物特异性标记，未标记的VX-710抑制类似物结合，EC（50）为0.75 μ M。VX-710还以浓度依赖性方式刺激钒酸盐可降解P-糖蛋白ATP酶活性2- 3倍，表观k（a）为0.1 μ M。这些数据表明，VX-710与P-糖蛋白的直接、高亲和力相互作用阻止了多药耐药肿瘤细胞中MDR 1基因产物的细胞毒性药物外排。
VX-710 or (S)-N-[2-Oxo-2-(3,4,5-trimethoxyphenyl)a cetyl]-piperidine-2-carboxylic acid 1,7-bis(3-pyridyl)-4-heptyl ester, a novel non-macrocyclic ligand of the FK506-binding protein FKBP12, was evaluated for its ability to reverse P-glycoprotein-mediated multidrug resistance in vitro. VX-710 at 0.5-5 mu M restored sensitivity of a variety of multidrug resistant cells to the cytotoxic action of doxorubicin, vincristine, etoposide or paclitaxel, including drug-selected human myeloma and epithelial carcinoma cells, and human MDR1 cDNA-transfected mouse leukemia and fibroblast cells. Uptake experiments showed that VX-710 at 0.5-2.5 mu M fully restored intracellular accumulation of [C-14]doxorubicin in multidrug resistant cells, suggesting that VX-710 inhibits the drug efflux activity of P-glycoprotein. VX-710 effectively inhibited photoaffinity labeling of P-glycoprotein by [H-3]azidopine or [I-125]iodoaryl azido-prazosin with EC(50) values of 0.75 and 0.55 mu M. Moreover, P-glycoprotein was specifically labeled by a tritiated photoaffinity analog of VX-710 and unlabeled VX-710 inhibited analog binding with an EC(50) of 0.75 mu M. VX-710 also stimulated the vanadate-inhibitable P-glycoprotein ATPase activity 2- to 3-fold in a concentration-dependent manner with an apparent k(a) of 0.1 mu M. These data indicate that a direct, high-affinity interaction of VX-710 with P-glycoprotein prevents efflux of cytotoxic drugs by the MDR1 gene product in multidrug resistant tumor cells.