Background
Methods
Chemicals
Animals
Pharmacokinetic (PK) study
Tissue distribution of [14C] DHA study
Mass balance and elimination of [14C] DHA study
Metabolic profile of [14C] DHA study
Protein binding of [14C] DHA study
HPLC-ECD method
Data evaluation
Results
Dose formulation analysis
Pharmacokinetics of [14C] DHA and unchanged DHA
14C-Dihydroartemisinin 3 mg/20 μCi/kg | Unchanged DHA 3 mg/kg | ||
---|---|---|---|
Parameters | Plasma Three-compartment | Blood Three-compartment | Plasma Two-compartment |
Cmax (ng/ml) | 3446 ± 396 | 3105 ± 430 | 1999 ± 507 |
Tmax (hr) | 0.02 | 0.02 | 0.02 |
AUCinf. (ng*h/ml) | 21014 ± 2718 | 52690 ± 4971 | 933.2 ± 125.4 |
t1/2 – alpha (hr) | 0.21 ± 0.05 | 0.18 ± 0.02 | 0.25 ± 0.04 |
t1/2 – beta (hr) | 15.49 ± 1.64 | 19.68 ± 3.88 | 1.03 ± 0.19 |
t1/2 – gamma (hr) | 75.57 ± 7.34 | 122.13 ± 8.73 | - |
CL (ml/hr/kg) | 144.63 ± 18.17 | 43.58 ± 30.70 | 3350.02 ± 423.37 |
Vss (ml/kg) | 8216 ± 828 | 10792 ± 3771 | 1658 ± 540 |
MRT (hr) | 57.00 ± 2.84 | 148.57 ± 90.24 | 0.49 ± 0.12 |
Cmax at first peak (ng/ml) | 3446 ± 396 (0.02 h) | 3105 ± 430 (0.02 h) | 1999 ± 507 (0.02 h) |
at second peak (ng/ml) | 2373 ± 195 (0.19 h) | 1557 ± 955 (0.94 h) | 1638 ± 468 (0.38 h) |
at third peak (ng/ml) | 863 ± 537 (1.70 h) | 978 ± 164 (2.00 h) | - |
at fourth peak (ng/ml) | 584 ± 65.5 (6.00 h) | 833 ± 99 (6.00) | - |
Tissue distribution of [14C] DHA
Parameters | 1 hr | 6 hr | 24 hr | 48 hr | 72 hr | 96 hr | 192 hr | Half-lives (hr) | Tissue AUC (ng·hr/g) | % of total AUC |
---|---|---|---|---|---|---|---|---|---|---|
Blood | 936.19 | 795.07 | 366.76 | 252.05 | 194.59 | 130.92 | 118.71 | 122.13 ± 8.73 | 45463 ± 6810 | 1.07 |
Plasma | 591.34 | 540.83 | 216.03 | 93.02 | 64.70 | 34.58 | 22.84 | 75.57 ± 7.34 | 20750 ± 1031 | 0.49 |
Brain | 578.68 | 326.57 | 160.11 | 127.38 | 138.35 | 119.67 | 133.52 | 72.21 ± 6.33 | 34189 ± 4885 | 0.81 |
Eyes | 279.20 | 239.95 | 64.78 | 46.86 | 51.58 | 98.06 | 121.94 | 75.22 ± 9.40 | 16519 ± 3048 | 0.39 |
Adrenals | 5614.75 | 3451.81 | 2034.62 | 3002.03 | 1630.57 | 1653.43 | 1543.60 | 92.48 ± 44.34 | 448390 ± 273502 | 10.59 |
Fat | 1391.80 | 218.23 | 80.24 | 62.09 | 61.62 | 43.52 | 84.86 | 131.69 ± 21.94 | 25253 ± 1490 | 0.60 |
Muscle | 698.14 | 499.29 | 171.74 | 163.19 | 174.04 | 92.20 | 115.14 | 64.45 ± 13.54 | 36371 ± 6994 | 0.86 |
Lungs | 1161.72 | 1040.99 | 398.52 | 305.34 | 303.10 | 236.70 | 221.64 | 86.19 ± 18.00 | 85916 ± 12999 | 2.03 |
Heart | 1512.27 | 1107.89 | 647.58 | 527.3 | 568.43 | 377.26 | 443.20 | 74.35 ± 25.84 | 104598 ± 24036 | 2.47 |
Spleen | 4387.40 | 3684.97 | 3061.40 | 4216.03 | 4446.65 | 4464.57 | 5170.22 | 114.02 ± 57.67 | 617495 ± 410768 | 14.58 |
Kidneys | 4097.87 | 4502.00 | 2535.56 | 2324.72 | 3286.27 | 1802.63 | 1819.89 | 77.90 ± 8.41 | 191781 ± 66422 | 4.53 |
Liver | 4594.85 | 3516.69 | 1403.29 | 1104.94 | 1503.74 | 842.90 | 1270.24 | 95.18 ± 37.90 | 172474 ± 85780 | 4.07 |
Stomach | 1366.22 | 932.33 | 464.21 | 342.30 | 337.48 | 229.69 | 218.27 | 72.58 ± 12.98 | 82125 ± 13428 | 1.94 |
Stomach Content | 36.03 | 6289.79 | 22.45 | 98.12 | 18.69 | 11.76 | 343.37 | 16.75 ± 16.30 | 22362 ± 98089 | 0.53 |
Large intestine (LI) | 1818.56 | 8394.43 | 1420.58 | 562.96 | 509.29 | 220.41 | 304.72 | 62.53 ± 43.59 | 70217 ± 25001 | 1.66 |
LI Content | 2914.59 | 35567.01 | 5127.28 | 2037.53 | 1426.62 | 491.66 | 1333.83 | 37.20 ± 6.08 | 588542 ± 104261 | 13.90 |
Small intestine (SI) | 8155.95 | 3014.58 | 821.84 | 335.42 | 389.72 | 235.61 | 304.12 | 49.99 ± 34.89 | 131264 ± 24631 | 3.10 |
SI Content | 46426.85 | 11387.30 | 2283.61 | 633.61 | 575.05 | 176.57 | 250.03 | 28.73 ± 9.05 | 1492988 ± 63053 | 35.25 |
Testis | 624.14 | 513.53 | 176.47 | 155.78 | 151.29 | 123.54 | 141.04 | 86.24 ± 40.68 | 48249 ± 6078 | 100.00 |
Total Amounts | 87186.55 | 86023.26 | 21457.06 | 16390.68 | 15831.77 | 11385.67 | 13961.18 | 69.03 ± 22.13 | 4234952.00 | 1.14 |
% total recovery | 34.57 | 34.10 | 8.51 | 6.50 | 6.28 | 4.51 | 5.53 |
Mass balance determination
Sample time (days) | Urine | Feces | Cage wash |
---|---|---|---|
0–0.04 | 4.18 ± 1.06 | ||
0.04–0.33 | 25.49 ± 1.98 | 1.52 ± 0.58 | |
0.33–1.0 | 14.01 ± 5.11 | 29.56 ± 11.75 | |
1–2 | 4.78 ± 1.30 | 6.14 ± 1.71 | |
2–3 | 1.63 ± 0.92 | 1.64 ± 0.78 | |
3–4 | 0.72 ± 0.19 | 0.78 ± 0.31 | |
4–5 | 0.57 ± 0.25 | 0.46 ± 0.09 | |
5–6 | 0.46 ± 0.37 | 0.15 ± 0.06 | |
6–7 | 0.16 ± 0.02 | 0.11 ± 0.05 | |
7–8 | 0.09 ± 0.04 | 0.04 ± 0.04 | |
Total | 52.09 ± 8.94 | 40.39 ± 13.22 | 1.88 ± 2.27 |
Ratio of excretion (urine/feces) | 1.37 ± 0.39 | ||
Balance (% of dose) | 94.36 ± 16.71 | ||
Elimination parameters | |||
Peak height (μg/rat) | 234.48 ± 33.74 | 292.39 ± 150.29 | |
Peak time (days) | 0.37 ± 0.06 | 0.67 ± 0.09 | |
Fast elimination phase (days) | 0.32 ± 0.07 | 0.35 ± 0.11 | |
Slow elimination phase (days) | 1.36 ± 0.25 | 1.29 ± 0.47 |
Urinary and faecal elimination
Metabolite profiles of [14C] DHA
Time | Total | Free fraction | Glucuronide conjugation | Other conjugation fraction |
---|---|---|---|---|
Plasma (n = 5) | ||||
0 | 0.00 | 0.00 | 0.00 | 0.00 |
0.017 | 3446.38 ± 396.35 | 2253.63 ± 537.31 | 403.87 ± 124.77 | 583.17 ± 155.52 |
0.083 | 2279.43 ± 213.77 | 925.14 ± 226.81 | 413.56 ± 69.46 | 589.70 ± 102.30 |
0.167 | 2090.86 ± 180.56 | 697.05 ± 88.04 | 480.62 ± 49.17 | 818.11 ± 88.12 |
0.25 | 1987.48 ± 378.69 | 685.05 ± 193.99 | 360.93 ± 65.83 | 684.75 ± 161.05 |
0.50 | 1489.58 ± 231.59 | 461.20 ± 56.83 | 305.81 ± 26.23 | 627.62 ± 59.19 |
0.75 | 654.49 ± 87.56 | 157.51 ± 28.11 | 131.60 ± 29.09 | 341.12 ± 66.12 |
1.0 | 514.20 ± 139.37 | 105.36 ± 55.10 | 122.76 ± 36.01 | 348.30 ± 125.91 |
1.5 | 419.48 ± 17.06 | 75.83 ± 9.85 | 74.17 ± 11.64 | 268.16 ± 14.76 |
2 | 589.26 ± 154.02 | 90.58 ± 33.58 | 110.13 ± 44.55 | 407.78 ± 112.11 |
3 | 466.65 ± 82.13 | 74.40 ± 16.25 | 80.73 ± 14.20 | 321.61 ± 69.08 |
6 | 584.41 ± 65.49 | 51.49 ± 15.64 | 49.34 ± 22.19 | 363.63 ± 162.04 |
8 | 481.59 ± 79.35 | 49.44 ± 13.34 | 57.72 ± 13.56 | 388.11 ± 72.73 |
24 | 253.62 ± 78.47 | 14.46 ± 3.15 | 20.04 ± 5.05 | 188.12 ± 47.66 |
48 | 79.17 ± 6.71 | 7.41 ± 3.24 | 8.55 ± 1.68 | 77.07 ± 14.36 |
72 | 64.38 ± 14.66 | 4.89 ± 1.83 | 4.31 ± 1.19 | 48.23 ± 12.74 |
96 | 35.26 ± 6.54 | 2.09 ± 1.39 | 2.25 ± 0.87 | 25.39 ± 7.32 |
192 | 13.48 ± 3.65 | 1.01 ± 0.92 | 0.76 ± 0.72 | 14.49 ± 1.62 |
AUC (ng·h/ml) | 21708 ± 2695 | 2188.2 ± 286.4 | 2177.6 ± 299.6 | 14193.6 ± 6838.7 |
t1/2 (hr) | 15.49 ± 1.64 | 4.01 ± 0.99 | 6.66 ± 1.04 | 20.25 ± 1.32 |
% of total | 100.2 ± 12.68 | 10.08 ± 1.32 | 10.03 ± 1.38 | 79.14 ± 8.22 |
Urine (n = 5) | ||||
0 | 0.00 | 0.00 | 0.00 | 0.00 |
0.04 | 37186.4 ± 8200.9 | 2253.63 ± 113.44 | 1611.28 ± 756.20 | 21546.88 ± 8219.11 |
0.33 | 228363.9 ± 20101.9 | 2693.12 ± 1293.63 | 8359.94 ± 1428.22 | 39881.66 ± 5139.02 |
1.0 | 123776.8 ± 38621.3 | 535.48 ± 339.64 | 732.58 ± 373.98 | 10784.80 ± 6618.73 |
2.0 | 42340.5 ± 8839.2 | 184.08 ± 70.94 | 97.20 ± 50.00 | 2003.77 ± 1218.99 |
3.0 | 14248.3 ± 6958.5 | 86.77 ± 56.02 | 29.60 ± 15.80 | 655.31 ± 605.27 |
4.0 | 6355.5 ± 1351.3 | 43.20 ± 26.91 | 17.54 ± 11.47 | 278.99 ± 208.65 |
5.0 | 4984.6 ± 1922.7 | 35.21 ± 39.14 | 12.70 ± 9.90 | 310.60 ± 306.84 |
6.0 | 4006.4 ± 2925.2 | 22.16 ± 28.94 | 7.77 ± 3.88 | 289.69 ± 338.11 |
7.0 | 1451.9 ± 149.2 | 9.75 ± 4.98 | 6.21 ± 2.92 | 147.26 ± 104.46 |
8.0 | 780.1 ± 344.4 | 5.77 ± 3.86 | 3.28 ± 1.66 | 91.76 ± 59.41 |
AUC (ng·h/ml) | 43624 ± 10214 | 2213.6 ± 933.4 | 5069.3 ± 928.2 | 35657.7 ± 9342.1 |
t1/2 (day) | 1.36 ± 0.25 | 1.25 ± 0.11 | 1.30 ± 0.16 | 1.64 ± 0.11 |
% of total | 98.43 ± 23.38 | 5.11 ± 1.45 | 12.22 ± 3.14 | 82.67 ± 3.96 |