A fluorometric screening assay for drug efflux transporter activity in the blood-brain barrier

被引:20
作者
Bachmeier, CJ [1 ]
Miller, DW [1 ]
机构
[1] Univ Nebraska, Med Ctr, Dept Pharmaceut Sci, Omaha, NE 68198 USA
关键词
BCECF; blood-brain barrier; MRP; P-gp;
D O I
10.1007/s11095-004-9016-0
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
Purpose. To examine the capability of a fluorometric assay to identify and characterize the drug efflux interactions of a broad spectrum of drug agents in an hi vitro model of the blood-brain barrier (BBB). Methods. Various concentrations of drug agent (1, 10, and 100 muM) were evaluated for their effect on the cellular accumulation of the P-glycoprotein (P-gp) probe R123 (3.2 muM), and the mixed P-gp and multidrug resistance-associated protein (MRP) probe, BCECF (1 muM), in bovine brain microvessel endothelial cell (BBMEC) monolayers. Drugs demonstrating a significant effect were further quantitated using an expanded concentration range and a nonlinear regression curve fit to determine the potency (IC50) and efficacy (Imax) of the drug for P-gp and/or MRP. Results. Several of the 36 therapeutic agents examined showed drug efflux transporter interactions in BBMEC monlayers. Melphalan and risperidone significantly enhanced the accumulation of R123 over control (1.47- and 1.82-fold, respectively) with resulting IC(50)s of 1.4 and 14.6 muM, respectively. Chlorambucil and valproic acid significantly enhanced the accumulation of BCECF compared to control monolayers (2.02- and 4.01-fold, respectively) With resulting IC50S of 146.1 and 768.5 muM, respectively. Conclusions. The current study demonstrates the feasibility of a fluorometric assay consisting of R123 and BCECF in assessing the drug efflux interactions of a variety of drugs in the BBB.
引用
收藏
页码:113 / 121
页数:9
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