Stephanie van Hoppe

27 ABCG2 & ABCB1 transport afatinib and restrict its oral availability and brain accumulation Figure 1. In vitro transport of afatinib. Transepithelial transport of afatinib (2 µM) was assessed in MDCK-II cells either non-transduced ( A, B ) or transduced with hABCB1 ( C, D ), hABCG2 ( E, F ), or mAbcg2 ( G, H ) cDNA. At T = 0 h afatinib was added to the donor compartment; thereafter at t = 2, 4 and 8 h the concentrations were measured and plotted as total amount (nmol) of transport (n = 3). B, D-H : Zosuquidar (5 µM) and/or Ko143 (5 µM) were added as indicated to inhibit hABCB1 and hABCG2 or mAbcg2, respectively. r , relative 0 2 4 6 8 0 0.5 1.0 1.5 2.0 Afatinib transport (nmol) hABCG2 + Zos 0 2 4 6 8 0 0.5 1.0 1.5 2.0 Time (h) Afatinib transport (nmol) mAbcg2 + Zos 0 2 4 6 8 0 0.5 1.0 1.5 2.0 Time (h) Afatinib transport (nmol) hABCG + KO143 + Zos 0 2 4 6 8 0 0.5 1.0 1.5 2.0 Time (h) Afatinib transport (nmol) mAbcg2 + KO143 + Zos 0 2 4 6 8 0.0 0.5 1.0 1.5 2.0 time (hours) Afatinib transport (nmol) Parental 0 2 4 6 8 0.0 0.5 1.0 1.5 2.0 time (hours) Afatinib transport (nmol) hABCB1 0 2 4 6 8 0.0 0.5 1.0 1.5 2.0 time (hours) Afatinib transport (nmol) AB BA Parental + Zosuquidar 0 2 4 6 8 0.0 0.5 1.0 1.5 2.0 time (hours) Afatinib transport (nmol) hABCB1 + Zosuquidar r = 1.3 r = 0.8 r = 7.6 r = 0.9 r = 5.6 r = 0.8 r = 23 r = 1.0 * ** *** *** *** A B C D E F G H AB BA AB BA AB BA Figure 1 - In vitro transport of af tinib. Transepithelial transport of fatinib (2 µM) was assessed in MDCK-II cells eith r non-transduced ( A, B ) r transduc with hABCB1 ( C, D ), hABCG2 ( E, F ), or mAbcg2 ( G, H ) cDNA. At T = 0 h afatinib was added to the donor comp rtment; her after at t = 2, 4 and 8 h the concentra ions were measured an plot ed as total m unt (nmol) of transport (n = 3). B, D-H : Zosuquidar (5 µM) and/o Ko143 (5 µM were added as ndicated to inhibit hABCB1 and hABCG2 o mAbcg2, respectively. r , relative transport ratio. BA (red squares) translocation from the basolateral to the apical compartment; AB (black circles), translocation from the apical to the basolateral compartment. Data are presented as the mean ± SD. Owing to high experimental reproducibility, in many cases the error bars are smaller than the symbols used to denote a data point. * , P < 0.05; ** , P < 0.01; *** , P < 0.001 compared with AB translocation.