Fellner Stephan, Bauer Björn, Miller David S, Schaffrik Martina, Fankhänel Martina, Spruss Thilo, Bernhardt Günther, Graeff Claudia, Färber Lothar, Gschaidmeier Harald, Buschauer Armin, Fricker Gert
Institute of Pharmacy, University of Regensburg, Regensburg, Germany.
J Clin Invest. 2002 Nov;110(9):1309-18. doi: 10.1172/JCI15451.
Paclitaxel concentrations in the brain are very low after intravenous injection. Since paclitaxel is excluded from some tumors by p-glycoprotein (p-gp), the same mechanism may prevent entry into the brain. In vitro, paclitaxel transport was examined in capillaries from rat brains by confocal microscopy using BODIPY Fl-paclitaxel. Western blots and immunostaining demonstrated apical expression of p-gp in isolated endothelial cells, vessels, and tissue. Secretion of BODIPY Fl-paclitaxel into capillary lumens was specific and energy-dependent. Steady state luminal fluorescence significantly exceeded cellular fluorescence and was reduced by NaCN, paclitaxel, and SDZ PSC-833 (valspodar), a p-gp blocker. Leukotriene C(4) (LTC(4)), an Mrp2-substrate, had no effect. Luminal accumulation of NBDL-cyclosporin, a p-gp substrate, was inhibited by paclitaxel. In vivo, paclitaxel levels in the brain, liver, kidney, and plasma of nude mice were determined after intravenous injection. Co-administration of valspodar led to increased paclitaxel levels in brains compared to monotherapy. Therapeutic relevance was proven for nude mice with implanted intracerebral human U-118 MG glioblastoma. Whereas paclitaxel did not affect tumor volume, co-administration of paclitaxel (intravenous) and PSC833 (peroral) reduced tumor volume by 90%. Thus, p-gp is an important obstacle preventing paclitaxel entry into the brain, and inhibition of this transporter allows the drug to reach sensitive tumors within the CNS.
静脉注射后,大脑中的紫杉醇浓度非常低。由于紫杉醇可被P-糖蛋白(P-gp)排除在某些肿瘤之外,相同的机制可能会阻止其进入大脑。在体外,使用BODIPY Fl-紫杉醇通过共聚焦显微镜检查了大鼠脑毛细血管中的紫杉醇转运。蛋白质免疫印迹和免疫染色显示,P-gp在分离的内皮细胞、血管和组织中呈顶端表达。BODIPY Fl-紫杉醇向毛细血管腔的分泌具有特异性且依赖能量。稳态管腔荧光显著超过细胞荧光,并被NaCN、紫杉醇和P-gp阻滞剂SDZ PSC-833(伐司朴达)降低。白三烯C4(LTC4)是一种多药耐药相关蛋白2(Mrp2)底物,没有影响。P-gp底物NBDL-环孢素的管腔蓄积受到紫杉醇的抑制。在体内,静脉注射后测定了裸鼠脑、肝、肾和血浆中的紫杉醇水平。与单一疗法相比,联合使用伐司朴达可提高脑中的紫杉醇水平。对于植入人脑U-118 MG胶质母细胞瘤的裸鼠,已证明其具有治疗相关性。虽然紫杉醇不影响肿瘤体积,但联合使用紫杉醇(静脉注射)和PSC833(口服)可使肿瘤体积缩小90%。因此,P-gp是阻止紫杉醇进入大脑的重要障碍,抑制这种转运蛋白可使药物到达中枢神经系统内的敏感肿瘤。