新型紫杉烷化合物NPB304及其协同维拉帕米逆转耐药的研究
Resistance reversal effect of a novel taxane compound NPB304 and its collaboration with verapamil
梅梅 1张翼 1任金红 1谢丹 1贾雨霏 1扈金萍 1李燕 1戴均贵 1陈晓光1
作者信息
- 1. 中国医学科学院、北京协和医学院药物研究所,北京100050
- 折叠
摘要
研究新型紫杉烷类化合物NPB304逆转肿瘤多药耐药的作用.MTT法测定化疗药物的IC50,Westernblotting方法分析P-糖蛋白(P-gp)的表达,分别通过罗丹明123 (Rh123)蓄积实验和分析试剂盒测定化合物对P-gp功能及P-gp ATPase活性的影响,采用分子对接预测化合物与P-gp结合能力的强弱,用MDCK Ⅱ和MDRl-MDCKⅡ细胞模型分析NPB304的跨膜转运.在KBV细胞中,NPB304具有多药耐药逆转作用;在MCF-7/paclitaxel细胞中,NPB304协同P-gp抑制剂维拉帕米增强逆转耐药的活性,10μmol·L-1维拉帕米与paclitaxel合用时逆转倍数为56.5倍,合用NPB304增加耐药逆转倍数;NPB304与维拉帕米合用时协同增加Rh123在耐药细胞中的蓄积,NPB304 (0～1 μmol·L-1)增强维拉帕米激活P-gp ATPase活性的作用;NPB304与P-gp的TM区存在疏水相互作用,与TM区A链的结合力较强;NPB304在较低浓度(0～1.5 μmol·L-1)时激活P-gp ATPase,发挥抑制P-gp功能的作用,但不具有明显的P-gp底物特征.NPB304通过抑制P-gp的功能活性发挥自身及协同维拉帕米逆转耐药的作用.
Abstract
The tumor multidrug resistance reversal effect of NPB304,a novel taxane,was studied.MTT assay was used to determine the IC 50 of chemotherapy drugs.Western blotting assay was applied to analyze the expression of P-glycoprotein (P-gp).The effect of compounds on the P-gp function and P-gp ATPase activity was determined by rhodamine 123 (Rh123) accumulation assay and analysis kit,respectively.Molecular docking was employed to predict the binding force between compounds and P-gp.Transmembrane transport of NPB304 was analyzed using MDCK Ⅱ and MDR1-MDCKⅡ cell model.NPB304 displayed multidrug resistance reversal effect on KBV cells and MCF-7/paclitaxel cells,NPB304 collaborative with P-glycoprotein (P-gp) inhibitors verapamil enhanced the reversal activity,specifically,10 μmol·L-1 verapamil in combination with paclitaxel reversed resistance by 56.5-fold,while combined with NPB304 increased the reversal fold; NPB304 synergistically increased Rh123 accumulation in the resistant cells when combined with verapamil,and NPB304 at 0-1 μmol·L-1 enhanced the ATPase activity activated by verapamil was observed.NPB304 existed the hydrophobic interactions with the TM regions of P-gp,and the binding force between NPB304 and the A chain of the TM region was stronger.P-gp ATPase activity assay demonstrated NPB304 at lower concentrations (0-1.5 μmol·L 1) could activate the P-gp ATPase,playing a role on inhibition of P-gp function.However,NPB304 did not have an obvious feature of P-gp substrate.NPB304 exerted itself and synergy with verapamil activity on reversing tumor resistance via inhibiting the P-gp function.
关键词
NPB304/肿瘤/耐药逆转/维拉帕米/P-糖蛋白Key words
NPB304/tumor/resistance reversal/verapamil/P-glycoprotein引用本文复制引用
基金项目
国家自然科学基金资助项目(81102469)
出版年
2014
NETL
NSTL
维普
万方数据