Article Figures & Data
Figures
- Fig. 1.
The rate of the uptake vs. the unbound concentration of pravastatin (A), rosuvastatin (B), repaglinide (C), and pitavastatin (D) in OATP1B1- and OATP1B3-overexpressing cells in buffer and human plasma. Total uptake (overexpressed cells) is depicted by circles, passive diffusion (control cells) is depicted by squares, and active uptake (the difference) is depicted by triangles.
- Fig. 2.
The plasma protein binding vs. the fold difference in the CLint value of pravastatin, rosuvastatin, repaglinide, and pitavastatin generated in buffer and human plasma for OATP1B1 and OATP1B3.
Tables
- TABLE 1
Liquid chromatography–tandem mass spectrometry methods for pravastatin, rosuvastatin, repaglinide, and pitavastatin
Analyte ESI Q1/Q3 Transition (m/z ratio) Gradient Profile [min (%B)] Flow Rate (1.0 ml/min) Declustering Potential Collision Energy Collision Cell Exit Potential Limit of Quantification Lower Upper nM nM Pitavastatin + 422.2/274.2 0.10 (5%)–0.40 (5%)–1.30 (85%)–1.60 (85%)–1.61 (5%)–2.00 (5%) 88.0 65.0 12.0 0.376 114 Repaglinide + 453.2/230.2 0.10 (5%)–0.40 (5%)–1.30 (85%)–1.60 (85%)–1.61 (5%)–2.00 (5%) 66.0 35.0 18.0 0.0610 250 Rosuvastatin + 482.3/258.3 0.40 (10%)–1.19 (90%)–1.60 (90%)–1.61 (10%)–2.00 (10%) 125 45.0 10.0 0.977 1000 Pravastatin − 423.1/303.0 0.10 (10%)–1.00 (80%)–1.40 (80%)–1.41 (10%)–1.80 (10%) −175 −22.0 −17.0 1.95 1000 ESI, electrospray ionization; m/z, mass-to-charge ratio; +/−, positive/negative ion mode.
- TABLE 2
The viability of the OATP1B1 and OATP1B3 cells in protein-free buffer and 100% human plasma
Viability in Protein-Free Buffer Viability in 100% Human Plasma Time in Buffer OATP1B1 Viability OATP1B3 Viability Time in Plasma OATP1B1 Viability OATP1B3 Viability min % % min % % 1 81.3 90.4 1 91.1 85.9 2 85.2 90.1 2 88.6 88.2 3 89.6 88.0 3 89.3 82.2 - TABLE 3
Human plasma protein binding values of the pravastatin, rosuvastatin, repaglinide, and pitavastatin (n = 3)
Compound Concentration fup μM Pravastatin 3 0.5157 ± 0.0726 200 0.5414 ± 0.0139 Rosuvastatin 1 0.2526 ± 0.0032 50 0.2499 ± 0.0538 Repaglinide 0.1 0.0224 ± 0.0011 1 0.0239 ± 0.0033 Pitavastatin 1 0.0074 ± 0.0006 10 0.0097 ± 0.0006 - TABLE 4
The Km,u, Vmax, Pdif,u, CLint, and percentage of active uptake values generated for pravastatin, rosuvastatin, repaglinide, and pitavastatin in OATP1B1 and OATP1B3 overexpressing cells in buffer and human plasma (n = 3)
CLint = Vmax/Km,u.
Compound Transporter Incubation Km,u Vmax Km,u (Reported in the Literature) Pdif,u CLint Active uptake μM pmol/min per milligram μM μl/min per milligram μl/min per milligram % Pravastatin OATP1B1 Buffer 84.9 ± 19.8 274 ± 29.1 85.7a, 104b, 109c 0.159 ± 0.0143 3.23 ± 0.827 95.3 Plasma 44.5 ± 12.8 224 ± 29.7 0.614 ± 0.0419 5.03 ± 1.59 89.1 Fold difference (Buffer/Plasma) 1.91 1.22 Fold difference (Plasma/Buffer) 3.86 1.56 OATP1B3 Buffer 57.5 ± 11.5 179 ± 14.5 228b 0.159 ± 0.0143 3.11 ± 0.672 95.1 Plasma 28.5 ± 7.02 145 ± 14.2 0.614 ± 0.0419 5.09 ± 1.35 89.2 Fold difference (Buffer/Plasma) 2.02 1.23 Fold difference (Plasma/Buffer) 3.86 1.63 Rosuvastatin OATP1B1 Buffer 22.2 ± 5.25 132 ± 14.5 9.31d, 13.0e, 13.1f 0.677 ± 0.0407 5.95 ± 1.55 89.8 Plasma 3.25 ± 1.41 50.0 ± 8.47 2.64 ± 0.116 15.4 ± 7.17 85.4 Fold difference (Buffer/Plasma) 6.83 2.64 Fold difference (Plasma/Buffer) 3.90 2.59 OATP1B3 Buffer 18.0 ± 1.74 156 ± 6.57 14.2g, 16.5e 0.677 ± 0.0407 8.67 ± 0.914 92.8 Plasma 5.61 ± 1.17 37.1 ± 3.60 2.64 ± 0.116 6.61 ± 1.52 71.5 Fold difference (Buffer/Plasma) 3.21 4.20 Fold difference (Plasma/Buffer) 3.90 0.762 Repaglinide OATP1B1 Buffer 1.07 ± 0.414 42.9 ± 9.50 1.36d 48.6 ± 1.78 40.1 ± 17.9 45.2 Plasma 0.0100 ± 0.00587 1.98 ± 0.501 84.2 ± 2.70 198 ± 126 70.2 Fold difference (Buffer/Plasma) 107 21.7 Fold difference (Plasma/Buffer) 1.73 4.94 OATP1B3 Buffer N/A N/A N/A 48.6 ± 1.78 N/A N/A Plasma N/A N/A 84.2 ± 2.70 N/A N/A Fold difference (Buffer/Plasma) N/A Fold difference (Plasma/Buffer) 1.73 Pitavastatin OATP1B1 Buffer 5.29 ± 0.984 111 ± 10.1 1.30h, 2.48d, 3.00i 6.33 ± 0.109 21.0 ± 4.35 76.8 Plasma 0.00855 ± 0.00413 5.69 ± 1.29 148 ± 18.9 665 ± 355 81.8 Fold difference (Buffer/Plasma) 619 19.5 Fold difference (Plasma/Buffer 23.4 31.7 OATP1B3 Buffer 8.80 ± 0.748 152 ± 7.46 2.60j, 3.25i 6.33 ± 0.109 17.3 ± 1.70 73.2 Plasma 0.0157 ± 0.0270 1.56 ± 1.54 148 ± 18.9 99.4 ± 192 40.2 Fold difference (Buffer/Plasma) 561 97.4 0.174 Fold difference (Plasma/Buffer) 23.4 5.75
