The pharmacokinetics of oral dihydroartemisinin and artesunate in healthy Thai volunteers.

The pharmacokinetics of oral dihydroartemisinin (DHA) following the dose of 2 and 4 mg/ kg body weight dihydroartemisinin (Twisinin, T-2 Program, Thailand) and 4 mg/kg body weight oral artesunate (AS; Guilin Pharmaceutical Works, Guangxi, China) were investigated in 20 healthy Thai volunteers (10 males, 10 females). All formulations were generally well tolerated. Oral DHA was rapidly absorbed from gastrointestinal tract with marked inter-individual variation. The pharmacokinetics of DHA following the two dose levels were similar and linearity in its kinetics was observed. Based on the model-independent pharmacokinetic analysis, median (95% CI) values for Cmax of 181 (120-306) and 360 (181-658) ng/ml were achieved at 1.5 hours following 2 and 4 mg/kg body weight dose, respectively. The corresponding values for AUC0-infinity, t1/2z, CL/f and Vz/f were 377 (199-1,128) vs 907 (324-2,289) ng.h/ml, 0.96 (0.70-1.81) vs 1.2 (0.75-1.44) hours, 7.7 (4.3-12.3) vs 6.6 (3.1-10.1) l/kg, and 90.5 (28.6-178.2) vs 6.6 (3.1-10.1) ml/min/kg, respectively (2 vs 4 mg/kg dose). Oral AS was rapidly biotransformed to DHA, which was detectable in plasma as early as 15 minutes of AS dosing. Following 4 mg/kg dose, median (95% CI) value for Cmax of 519 (236-284) ng/ml was achieved at 0.7 (0.25-1.5) hours. AUC0-infinity, and t1/2z were 657 (362-2,079) ng.h/ml, 0.74 (0.34-1.42) hours, respectively. Cmax of DHA following oral AS were significantly higher, but total systemic exposure was greater following oral DHA at the same dose level (4 mg/kg body weight). There was no significant sex difference in pharmacokinetics of DHA.

Pharmacokinetics of prophylactic mefloquine.

The pharmacokinetics of the prophylactic dose of mefloquine (Lariam: 500 mg every 4 weeks, with a loading dose of 750 mg on the first week) was studied in six healthy Thai male volunteers. Mefloquine was well tolerated during the study period of 16 weeks. The only side-effects found were nausea and diarrhea in 2 volunteers after the first dose of mefloquine. The mean minimum concentration of mefloquine at steady state ranged from 290 to 460 ng/ml. The maximum concentration on week 16 after the last dose was 1558 +/- 48 ng/ml at the mean time of 38 +/- 19 hours. The other pharmacokinetic parameters obtained were: absorption half life = 6.6 +/- 3.0 hours; distribution = 5.1 +/- 3.1 days; terminal half life = 12.9 +/- 2.2 days; apparent volume of distribution = 10.5 +/- 2.3 l/kg; area under the concentration-time curve = 26.9 +/- 2.2 mg/dl. Although this prophylaxis regimen is ideal when considering the compliance, the minimum concentration obtained was much too low for optimum therapeutic concentration. We therefore suggest that weekly prophylaxis schedule should be a better regimen as the difference between minimum and maximum mefloquine concentration would be smaller.

Plasma quinine levels in patients with falciparum malaria when given alone or in combination with tetracycline with or without primaquine.

Quinine has been an effective drug for severe chloroquine-resistant falciparum malaria. However, there has been a decline in the sensitivity of Plasmodium falciparum to quinine. In 1978-1979 the cure rate was 94% compared to 86% in 1979-1980 and 76% in 1980-1981. The combination of quinine and tetracycline has improved the cure rate to 95-100%. However, the mechanism responsible for this has not been identified. We have compared plasma quinine levels on day 2, day 5 and day 7 (before and at 2 hours after dosing) in twenty-one patients with acute falciparum malaria who were treated with quinine alone (8 patients) or quinine in combination with tetracycline (8 patients) or quinine with tetracycline and primaquine (5 patients). All patients who received combination of quinine and tetracycline with or without primaquine responded well to the treatment with no recrudescence. Two patients who were treated with quinine alone had RI responses. Plasma quinine concentrations from the quinine alone group were significantly lower than those obtained from combination groups on days 2, 5 and 7. The minimal plasma quinine levels from quinine alone group were all lower than MIC, ie below 10 micrograms/ml while those obtained from the combination group were higher than MIC for 7 days. The results from the present study suggest that tetracycline has influence on the maintenance of plasma quinine levels above MIC throughout the treatment period. Therefore, this must be one possible explanation for the better cure rate.

Determination of quinine and quinidine in biological fluids by high performance liquid chromatography.

A simple, specific, rapid and sensitive High Performance Liquid Chromatography (HPLC) method has been developed to measure plasma level of quinine and quinidine. The drugs were extracted successively from plasma at basic pH with chloroform and quantified on a revers-phase Z-module C18 HPLC column with fluorescence detector (excitation 340 nm, emission 425 nm). The isocratic mobile phase used was the mixture of 0.05 M ammonium formate and acetonitrile (93.5:6.5, v/v), adjusted pH to 2.0 with ortho-phosphoric acid. The limits of quantitation for these compounds were as low as 4 ng/ml of plasma, using a 0.25 ml specimen. Calibration curves were linear (R squared = 0.9994) in the range 0-7,000 ng/ml. An interassay reproducibility was 6.8%, 0.3% and 1.2% at the concentrations of 250 ng, 4,000 ng and 8,000 ng of quinine, respectively. Inter-assay coefficient of variation of quinidine was 1.8%, 2.7% and 3.7% at the concentrations of 250 ng, 1,500 ng and 3,000 ng, respectively. Recovery of quinine and quinidine were 76% and 81%, respectively. The method has been used in the analysis of quinine and quinidine in healthy volunteers receiving quinine or quinidine intravenously. The method is now being used to assay samples from field studies with satisfactory results.

Determination of mefloquine in biological fluids using high performance liquid chromatography.

A simple, specific and sensitive High Performance Liquid Chromatography (HPLC) method for determination of whole blood of mefloquine has been developed. WR 184806 was used as internal standard, using a two step extraction procedure followed by revers phase HPLC. Acetonitrile and dichloromethane were used as extraction solvents. Octanesulphonic acid was used as an ionpairing reagent. Detection of extracted mefloquine and internal standard was achieved at 222 nm. Calibration curves for mefloquine in whole blood showed linearity with correlation coefficients of 0.9999. The limitation of detection using a 1 ml sample was 50 ng/ml. Recovery of mefloquine varied from 61% to 81%. Due to the very similar behavior of the internal standard during extraction, changes in recovery are of minor importance. Good accuracy and precision were obtained (intra-assay coefficient of variation ranged between 1.8% and 5%; inter-assay coefficient of variation were less than 10% at 100 ng/ml and less than 6% at 1,000 ng/ml). The assay employs a rapid and simple two-step extraction which requires a small sample volume. The low limit of detection of mefloquine and the short retention time make the method suitable for routine analysis of mefloquine.