Rosuvastatin Pharmacokinetics in Caucasian and Asian Subjects Residing in the United States

Online Resource 1: Supplementary Methods

Blood sampling

Venous blood samples (4 mL) for rosuvastatin, N-desmethyl rosuvastatin, and rosuvastatin lactone assay were taken before and at 0.5, 1, 2, 3, 4, 5, 6, 8, 10, 12, 18, 24, 30, 36, 48, 54, 60, and 72 h after rosuvastatin administration. Samples were collected into tubes containing lithium-heparin anticoagulant and centrifuged within 30 min; plasma was then harvested and mixed 1:1 with sodium acetate buffer, 0.1 mol/L (pH 4.0) and stored at –70°C until assayed.

Determination of plasma rosuvastatin, N-desmethyl rosuvastatin, and rosuvastatin lactone

Plasma samples were analyzed for rosuvastatin, N-desmethyl rosuvastatin, and rosuvastatin lactone by use of a method (high-performance liquid chromatography with tandem mass spectrometric detection) developed and validated at AstraZeneca (Wilmington, DE, US) (data on file). The method was established and validated at Covance Laboratories (Indianapolis, IN, US), which provided sample analysis for this trial. A robotic liquid-handling system was used to perform the sample preparation in a 96-well format. Plasma proteins were precipitated via simple protein precipitation and filtration. Analysis of the filtrate was accomplished by multiple-reaction monitoring via positive electrospray ionization-tandem mass spectrometric detection.

The lower limit of quantitation for rosuvastatin was 0.050 ng/mL. The upper limit was 100.0 ng/mL and was extended by dilution. The lower limit of quantitation for N-desmethyl rosuvastatin and rosuvastatin-lactone was 0.125 ng/mL; the upper limit was 25.0 ng/mL.

The accuracy and precision of the bioanalytical method were ensured on the basis of the results for spiked plasma quality control samples, which were assayed on each day of trial sample analysis. For rosuvastatin, accuracy averaged 100.0% (14.8% relative standard deviation [RSD]) at 0.25 ng/mL, 104.5% (5.7% RSD) at 6.25 ng/mL, and 105.2% (5.8% RSD) at 75.0 ng/mL. ForN-desmethyl rosuvastatin, accuracy averaged 103.4% (7.4% RSD) at 0.625 ng/mL, 105.5% (10.5% RSD) at 2.00 ng/mL, and 101.5% (7.0% RSD) at 20.0 ng/mL. For rosuvastatin-lactone, accuracy averaged 97.8% (9.7% RSD) at 0.625 ng/mL, 97.5% (9.5% RSD) at
2.00 ng/mL, and 98.5% (13.1% RSD) at 20.0 ng/mL.

Derivation of pharmacokinetic parameters

The apparent t1/2 was calculated as 0.693/z where z is the terminal elimination rate constant calculated by log-linear regression of the terminal portion of the plasma concentration-time curve. AUC(0–t) was determined by use of the linear trapezoidal rule, and AUC was determined as AUC(0–t) + Clast/z (where Clast is the last measurable plasma concentration).

Online Resource 2: Supplementary Tables
Supplementary Table 1 Dietary characteristics by ethnic groups

Chinese
n=26 / Filipino
n=27 / Asian-Indian
n=26 / Korean
n=26 / Vietnamese
n=26 / Japanese
n=27 / Caucasian
n=26
Daily caloric intake (kcal)
LSmean (SD) / 2,835
(1,334) / 2,663
(817.6) / 2,674
(1,153) / 2,696
(965.0) / 2,728
(883.1) / 2,244
(888.0) / 3,050
(1,103)
Differencea / -215.1 / -386.2 / -375.8 / -353.7 / -321.5 / -805.3
90% CI / -690.1–259.9 / -861.1–88.78 / -850.7–99.19 / -829.6–121.2 / -801.1–158.2 / -1,280– -330.4
% total fatb
LSmean (SD) / 36.44
(8.30) / 33.09
(8.79) / 33.71
(5.70) / 32.59
(9.15) / 30.08
(9.69) / 31.37
(8.86) / 34.57
(7.44)
Differencea / 1.87 / -1.48 / -0.86 / -1.97 / -4.49 / -3.19
90%CI / -1.96–5.71 / -5.31–2.35 / -4.69–2.98 / -5.81–1.86 / -8.3– -0.62 / -7.03–0.64
% saturated fatb
LSmean (SD) / 10.56
(3.20) / 10.30
(3.92) / 10.81
(3.04) / 9.92
(4.12) / 8.99
(3.14) / 8.08
(3.55) / 10.70
(2.40)
Differencea / -0.14 / -0.40 / 0.11 / -0.78 / -1.71 / -2.62
90% CI / -1.69–1.41 / -1.95–1.16 / -1.44–1.67 / -2.33–0.77 / -3.28– -0.15 / -4.17– -1.07
% carbohydrateb
LSmean (SD) / 46.01
(10.55) / 49.98
(10.15) / 50.26
(7.08) / 51.43
(10.39) / 54.21
(10.92) / 50.07
(11.06) / 49.14
(10.32)
Differencea / -3.13 / 0.85 / 1.12 / 2.29 / 5.07 / 0.93
90% CI / -7.78–1.53 / -3.80–5.50 / -3.53–5.77 / -2.36–6.94 / 0.37–9.77 / -3.72–5.58
% proteinb
LSmean (SD) / 16.46
(4.20) / 15.78
(3.86) / 14.96
(3.55) / 15.86
(3.60) / 14.61
(3.42) / 16.61
(5.03) / 14.52
(4.43)
Differencea / 1.94 / 1.26 / 0.44 / 1.34 / 0.09 / 2.09
90% CI / 0.08–3.80 / -0.60–3.12 / -1.42–2.30 / -0.52–3.20 / -1.79–1.96 / 0.23–3.95
Daily cholesterol intake (mg)
LSmean (SD) / 409.2
(305.4) / 357.2
(193.1) / 257.6
(126.8) / 318.2
(242.3) / 328.4
(205.3) / 312.3
(232.0) / 485.7
(469.9)
Differencea / -76.44 / -128.5 / -228.1 / -167.5 / -157.3 / -173.4
90%CI / -201.9–
48.98 / -253.9–
-3.10 / -353.5–
-102.7 / -292.9–
-42.09 / -284.0–
-30.64 / -298.8–
-47.94

aDifference of Asian ethnic group to Caucasian subjects derived from least squares mean (ethnic group effect), bPercent of total daily caloric intake
CI confidence interval, LSmean least squares mean, SD standard deviation

Supplementary Table2: Geometric Mean Cmax and AUC(0-t) by pooled ethnic group and genotype

Polymorphism / Genotype / Cmax: Geometric Mean (95% CI) / AUC(0-t): Geometric Mean (95% CI)
Pooled-
Asiana
n=100 / Asian-Indian
n=16 / Caucasian
n=22 / Pooled-
Asiana
n=100 / Asian-Indian
n=16 / Caucasian
n=22
SLCO1B1: T521>C
(Val174Ala) / T\T
T\C
C\C / 19.67
(17.48, 22.13)
27.90
(22.34, 34.85)
– / 16.92
(13.04, 21.96)
–
– / 9.84
(7.52 12.88)
16.99
(11.10, 26.01)
16.30
(5.75, 46.24) / 181.1
(163.8, 200.2)
247.9
(205.2, 299.4)
– / 164.0
(131.4, 204.7)
–
– / 98.4
(78.3, 123.7)
165.7
(115.4, 237.9)
147.0
(60.6, 356.5)
SLCO1B1:
A388>G
(Asn130Asp) / A\A
A\G
G\G / 20.65
(13.68, 31.18)
20.56
(17.00, 24.85)
21.69
(18.85, 24.97) / 15.05
(8.73, 25.96)
20.42
(13.89, 30.03)
13.06
(7.57, 22.54) / 11.05
(7.68, 15.90)
11.05
(7.68, 15.89)
15.06
(8.73, 25.97) / 189.3
(133.7, 267.9)
182.4
(155.4, 214.1)
201.4
(178.8, 226.7) / 144.3
(91.1, 228.5)
199.7
(144.3, 276.5)
125.7
(79.4, 199.1) / 116.4
(85.7, 158.1)
98.9
(72.8, 134.4)
161.1
(101.7, 255.2)
ABCG2:
C421>A(Gly141Lys) / C\C
C\A
A\A / 17.46
(15.11, 20.18)
24.14
(20.79, 28.02)
44.56
(28.33, 70.06) / 16.05
(12.25, 21.03)
15.20
(5.53, 41.81)
39.60
(14.40, 108.93) / 11.56
(8.90, 15.01)
11.97
(8.17, 17.55)
– / 159.1
(140.9, 179.6)
222.4
(196.3, 252.1)
387.8
(265.2, 567.3) / 160.4
(127.8, 201.3)
159.0
(67.9, 372.1)
231.0
(98.7, 540.6) / 111.4
(89.4, 138.7)
124.9
(90.6, 172.3)
–

aThePooled-Asian group comprised all Chinese, Filipino, Japanese, Korean and Vietnamese subjects

AUC(0-t)area under the plasma concentration-time curve from time 0 to the time of the last quantifiable concentration,CI confidence interval, Cmaxmaximum plasma concentration

Online Resource 3: Supplementary figures

Supplementary Fig.1Box-and-whisker plots of rosuvastatin AUC(0–t) for Asian and Caucasian subjects.

The upper and lower edges of the box represent the 75th and 25th percentiles, respectively. The horizontal line through the box represents the median and the upper and lower whiskers represent the 95th and 5th percentiles, respectively, whilst the crosses represent the extreme values.

AUC(0–t)area under the plasma concentration-time curve from time 0 to the time of the last quantifiable concentration

Supplementary Fig. 2Box-and-whisker plots of rosuvastatin Cmax for Asian and Caucasian subjects.

Cmaxmaximum plasma concentration

Supplementary Fig. 3Box-and-whisker plots of N-desmethyl rosuvastatin AUC(0–t) for Asian and Caucasian subjects.

AUC(0–t)area under the plasma concentration-time curve from time 0 to the time of the last quantifiable concentration

Supplementary Fig. 4Box-and-whisker plots of N-desmethyl rosuvastatin Cmax for Asian and Caucasian subjects.

Cmax maximum plasma concentration