Article Figures & Data
Figures
- FIG. 1.
(A) [3H]raltegravir (1 μM) accumulation in CEM, CEMVBL100, and CEMVBL100 cells treated with 300 nM tariquidar. (B) [3H]saquinavir (1 μM) accumulation in CEM, CEMVBL100, and CEMVBL100 cells treated with 300 nM tariquidar. Data in A and B are expressed as mean CARs (n = 4 biological replicates; n = 4 experimental replicates per biological replicate) ± SD. *, P < 0.05; **, P < 0.01; ***, P < 0.001. (C) Apparent permeability of [3H]raltegravir and [3H]digoxin in the A-to-B (apical-to-basolateral) and B-to-A (basolateral-to-apical) directions across the Caco-2 transwell membrane, with and without the presence of 300 nM tariquidar. Data are expressed as mean apparent permeabilities (×10−6 cm/s; n = 3 experimental replicates) ± SD. *, P < 0.05; **, P < 0.01; ***, P < 0.001.
- FIG. 2.
(A) SLC22A6- and H2O-injected oocyte uptake of [3H]raltegravir over a 4-h incubation period. (B) SLC22A6- and H2O-injected oocyte uptake of [3H]tenofovir over a 4-h incubation period. (C) SLC22A6- and H2O-injected oocyte uptake of [3H]aminohippuric acid over a 4-h incubation period. Data in A, B, and C are expressed as mean drug accumulations (pmol/oocyte) (n = 5 experimental replicates from one biological replicate) ± standard errors (SE). (D) Concentration dependency of the uptake of [3H]raltegravir by SLC22A6. (E) Concentration dependency of the uptake of [3H]tenofovir by SLC22A6. Data in D and E are expressed as mean rates of [3H]tenofovir transport (pmol/oocyte/h) (n = 4 experimental replicates from one biological replicate) ± SE.
- FIG. 3.
(A) Accumulation of 1 μM [3H]raltegravir in SLC22A6- and H2O-injected oocytes with and without the addition of 300 μM tenofovir. (B) Accumulation of 1 μM [3H]tenofovir in SLC22A6- and H2O-injected oocytes with and without the addition of 300 μM raltegravir. Data in A and B are expressed as mean drug concentrations per oocyte (pmol/oocyte) (n = 5 experimental replicates from one biological replicate) ± SE. *, P < 0.05; **, P < 0.01; ***, P < 0.001. (C) Determination of IC50 for inhibition of 1 μM [3H]raltegravir SLC22A6 transport by tenofovir. Data are expressed as mean [3H]raltegravir oocyte concentrations (pmol/oocyte) (n = 5 experimental replicates from one biological replicate) ± SE. (D) Determination of IC50 for inhibition of 1 μM [3H]tenofovir SLC22A6 transport by raltegravir. Data are expressed as mean [3H]tenofovir oocyte concentrations (pmol/oocyte) (n = 5 experimental replicates from one biological replicate) ± SE.
- FIG. 4.
(A) [3H]raltegravir (1 μM) accumulation in peripheral blood mononuclear cells alone or treated with 300 nM tariquidar, 1 mM probenecid, or 1 mM glycyl sarcosine. (B) [3H]raltegravir (1 μM) accumulation in primary renal proximal tubule cells alone or treated with 1 mM probenecid or 100 μM tenofovir. (C) [3H]tenofovir (1 μM) accumulation in primary renal proximal tubule cells alone or treated with 1 mM probenecid or 100 μM raltegravir. (D) [3H]aminohippuric acid (1 μM) accumulation in primary renal proximal tubule cells alone or treated with 1 mM probenecid. Data in A, B, C, and D are expressed as mean CARs (n = 3 experimental replicates) ± SD. *, P < 0.05; **, P < 0.01; ***, P < 0.001. (E) Relative abundance of transporter RNA in cultured renal proximal tubule cells compared to transporter RNA in whole kidney (log percent) (n = 1 experimental replicate).
Tables
- TABLE 1.
Accumulation of 1 μM raltegravir and various positive-control compounds in oocytesa
Transporter Drug Mean drug concn (pmol/oocyte) ± SD RNA/H2O ratio (P value) RNA-injected oocytes H2O-injected oocytes SLCO1A2 RAL 0.39 ± 0.14 0.35 ± 0.07 1.11 (0.09) E3S 1.48 ± 0.63 0.27 ± 0.09 5.48 (<0.01) SLCO1B1 RAL 0.41 ± 0.05 0.38 ± 0.05 1.08 (0.09) E3S 2.68 ± 1.55 0.34 ± 0.17 7.97 (<0.01) SLCO1B3 RAL 0.63 ± 0.19 0.65 ± 0.13 0.97 (0.09) E3S 0.56 ± 0.23 0.21 ± 0.03 2.60 (<0.01) SLC22A6 RAL 0.44 ± 0.12 0.20 ± 0.03 2.22 (<0.01) AHA 9.39 ± 2.64 0.17 ± 0.04 56.9 (<0.01) SLC10A1 RAL 0.20 ± 0.03 0.20 ± 0.03 1.03 (0.57) TCA 0.28 ± 0.10 0.08 ± 0.05 3.51 (<0.01) SLC15A1 RAL 0.26 ± 0.12 0.17 ± 0.03 1.52 (<0.01) GLY 0.45 ± 0.13 0.11 ± 0.05 4.22 (<0.01) SLC15A2 RAL 0.19 ± 0.04 0.17 ± 0.03 1.08 (0.16) GLY 0.29 ± 0.19 0.11 ± 0.05 2.69 (<0.01) SLC22A1 RAL 0.21 ± 0.03 0.17 ± 0.04 1.21 (0.06) TEA 0.34 ± 0.06 0.17 ± 0.04 1.99 (<0.01) ↵a Results are expressed as the mean drug concentrations per oocyte (pmol/oocyte) (n ≥ 2 biological replicates; n ≥ 4 experimental replicates per biological replicate) ± standard deviations. Also shown are the ratios of drug accumulation between transporter RNA-injected and H2O-injected oocytes. RAL, raltegravir; E3S, estrone-3-sulfate; AHA, amminohippuric acid; TCA, taurocholic acid; GLY, glycyl sarcosine; TEA, tetraethyl ammonium.
