Article Figures & Data
Figures
- Fig. 1.
Expression efficacies of CYPs and their activities to hydroxylize tolbutamide. (A) Expression efficacies evaluated by Western blotting. The first line (COS-7) was transfected by the mock agent, and the expression efficacy was adjusted by the internal transfection control protein (Gaussia princeps luciferase, Glu). Glyceraldehyde-3-phosphate dehydrogenase is a housekeeping protein; (B) the relative tolbutamide hydroxylase activities of mutant CYP2C9 alleles in COS-7 cells were compared with that of wild-type CYP2C9*1 (as 100%). Significantly different from wild-type CYP2C9, *P < 0.05, **P < 0.01, respectively.
- Fig. 2.
(A) Plasma concentration-time curves of tolbutamide in healthy subjects with genotype of CYP2C9*1/*1 (■) (n = 7), CYP2C9*1/*3 (●) (n = 7), CYP2C9*1/*13 (▲) (n = 2), CYP2C9*3/*3 (*), and CYP2C9*3/*13 (□) after a single oral dose of 500 mg tolbutamide. (B) Plasma concentration-time curves of 4-hydroxytolbutamide in healthy subjects with the corresponding genotypes.
- Fig. 3.
(A) Plasma concentration-time curves of tolbutamide in healthy subjects with genotypes of CYP2C9*1/*1 (■) (n = 7), CYP2C9*1/*16 (●) (n = 2), CYP2C9*1/*19 (▲), CYP2C9*1/*27 (□), CYP2C9*1/*29 (○), and CYP2C9*1/*34 (*) after a single oral dose of 500 mg tolbutamide. (B) Plasma concentration-time curves of 4-hydroxytolbutamide in healthy subjects with the corresponding genotypes.
- Fig. 4.
(A) Plasma concentration-time curves of tolbutamide in healthy subjects with genotype CYP2C9*1/*1 (■) (n = 7), CYP2C9*1/*40 (●), and CYP2C9*1/*41 (▲) after a single oral dose of 500 mg tolbutamide. (B) Plasma concentration-time curves of 4-hydroxytolbutamide in healthy subjects with the corresponding genotypes. (C) Plasma concentration-time curves of tolbutamide in healthy subjects with genotype CYP2C9*1/*1 (■) (n = 7), CYP2C9*1/*42 (●), CYP2C9*1/*45 (▲), CYP2C9*1/*46 (*), and CYP2C9*1/*48 (□) after a single oral dose of 500 mg tolbutamide. (D) Plasma concentration-time curves of 4-hydroxytolbutamide in healthy subjects with the corresponding genotypes.
- Fig. 5.
Correlations between in vitro enzymatic activity and in vivo activity (t1/2 of tolbutamide).
- Fig. 6.
The amount of urinary 4-hydroxytolbutamide and carboxytolbutamide of 2C9 variants excreted over 24 hours compared with that of wild-type 2C9*1/*1.
- Fig. 7.
(A) Average serum glucose concentration-time profiles after single oral administration of 75 g dextrose in seven normal healthy subjects with CYP2C9*1/*1 (■) (n = 7) genotypes and seven subjects with CYP2C9*1/*3 (●) (n = 7) genotypes. (B) Average delta serum glucose level from the baseline measured before oral intake of 75 g dextrose in subjects with CYP2C9*1/*1 (■) (n = 7) and CYP2C9*1/*3 (●) (n = 7) genotypes.
Tables
- TABLE 1
Demographic characteristics and CYP2C9 genotypes of 30 subjects aged 21∼22 years old
Genotype Number Gender Body Weight (kg) BMI Male Female CYP2C9*1/*1 7 3 4 57.9 ± 13.3 21.31 ± 4.47 CYP2C9*1/*3 7 4 3 53.3 ± 10.3 19.71 ± 2.92 CYP2C9*1/*13 1 1 44 18.31 CYP2C9*3/*3 1 1 68 22.56 CYP2C9*3/*13 1 1 53 20.20 CYP2C9*1/*16 1 1 80 27.68 CYP2C9*1/*19 1 1 52.5 19.40 CYP2C9*1/*27 1 1 65 20.52 CYP2C9*1/*29 1 1 50 19.05 CYP2C9*1/*34 1 1 65 22.49 CYP2C9*1/*40 1 1 47 19.07 CYP2C9*1/*41 1 1 54 18.47 CYP2C9*1/*42 1 1 43 19.15 CYP2C9*1/*45 1 1 65 20.02 CYP2C9*1/*46 1 1 60 21.37 CYP2C9*1/*48 1 1 50 21.16 BMI, body mass index.
- TABLE 2
Tolbutamide hydroxylase activities of mutant CYP2C9 alleles in COS-7 cells compared with that of wild-type CYP2C9*1
Genotype Percentage of Matching with Wild-Type Genotype Percentage of Matching with Wild-Type CYP2C9*1 100.00% ± 10.18% CYP2C9*34 24.34% ± 0.39%** CYP2C9*3 10.73% ± 1.90%** CYP2C9*40 125.78% ± 2.60%** CYP2C9*13 1.28% ± 0.48%** CYP2C9*41 97.62% ± 6.11% CYP2C9*16 3.45% ± 0.48%** CYP2C9*42 0.32% ± 0.24%** CYP2C9*19 31.11% ± 7.43%** CYP2C9*45 0.82% ± 0.01%** CYP2C9*27 114.99% ± 5.38%** CYP2C9*46 29.06% ± 1.35%** CYP2C9*29 109.14% ± 2.60% CYP2C9*48 38.22% ± 1.27%** Values are mean ± SD. *,**Indicate significant difference from wild-type CYP2C9 at P < 0.05 and P < 0.01, respectively.
Genotype AUC0→∞ AUC0→t Cmax t1/2 CL/F mg/L × h mg/L × h mg/L h L/h CYP2C9*1/*1 970.61 ± 152.14 933.90 ± 149.02 65.94 ± 11.68 8.14 ± 1.24 0.528 ± 0.094 CYP 2C9*1/*3 1722.88 ± 379.50** 1666.49 ± 377.69** 77.34 ± 11.95 13.73 ± 1.80** 0.301 ± 0.058** CYP 2C9*1/*13 2636.63 2729.30 95.42 16.92 0.183 CYP 2C9*3/*3 6639.49 6393.57 77.02 52.71 0.075 CYP 2C9*3/*13 8792.82 8462.01 103.72 57.53 0.057 CYP 2C9*1/*16 1269.659 1322.83 71.53 12.30 0.378 CYP 2C9*1/*19 1451.51 1375.93 86.33 10.66 0.344 CYP 2C9*1/*27 853.5 814.95 70.72 7.30 0.586 CYP 2C9*1/*29 1015.49 992.06 71.58 8.29 0.492 CYP 2C9*1/*34 1846.61 1808.14 75.62 14.34 0.271 CYP 2C9*1/*40 756.18 729.97 42.13 6.60 0.661 CYP 2C9*1/*41 1038.72 985.34 65.99 7.69 0.481 CYP 2C9*1/*42 2094.06 1919.56 87.16 15.85 0.239 CYP 2C9*1/*45 1654.64 1563.04 71.12 13.41 0.302 CYP 2C9*1/*46 1813.07 1754.54 94.98 12.00 0.276 CYP 2C9*1/*48 3197.57 3168.09 79.45 23.54 0.156 Values are mean ± SD. *,**Indicate significant difference from wild-type CYP2C9 at P < 0.05 and P < 0.01, respectively.
Genotype AUC0→∞ AUC0→t Cmax t1/2 mg/L × h mg/L × h mg/L h CYP2C9*1/*1 5.812 ± 1.219 5.579 ± 1.253 0.359 ± 0.090 7.642 ± 2.285 CYP 2C9*1/*3 5.176 ± 0.904 5.042 ± 0.858 0.252 ± 0.035* 9.237 ± 3.052 CYP 2C9*1/*13 7.608 7.426 0.301 10.025 CYP 2C9*3/*3 2.653 2.441 0.065 211.759 CYP 2C9*3/*13 3.976 3.908 0.072 43.997 CYP 2C9*1/*16 3.551 3.542 0.177 5.553 CYP 2C9*1/*19 7.981 6.749 0.360 16.716 CYP 2C9*1/*27 7.506 6.414 0.396 11.635 CYP 2C9*1/*29 9.643 9.151 0.514 10.175 CYP 2C9*1/*34 5.947 5.860 0.252 8.453 CYP 2C9*1/*40 5.47 4.746 0.389 7.092 CYP 2C9*1/*41 7.534 7.013 0.409 8.351 CYP 2C9*1/*42 5.938 5.839 0.242 10.033 CYP 2C9*1/*45 5.975 5.901 0.221 7.773 CYP 2C9*1/*46 6.605 6.597 0.298 4.812 CYP 2C9*1/*48 7.402 7.268 0.217 19.589 Values are mean ± SD. *,**Indicate significant difference from wild-type CYP2C9 at P < 0.05 and P < 0.01, respectively.
