J.F. Sutra

Enhancement of moxidectin bioavailability in lamb by a natural flavonoid: quercetin

Moxidectin is an antiparasitic drug widely used in cattle, sheep and companion animals. Due to the involvement of P-glycoprotein (P-gp) and cytochrome P450 3A in the metabolism of moxidectin, we studied the influence of various P-gp interfering agents (ivermectin, quercetin and ketoconazole) on the metabolism of 14C moxidectin in cultured rat hepatocytes over 72 h. This in vitro study allowed selection of compounds which are able to increase the moxidectin bioavailability in lambs. From this, the modulation of moxidectin pharmacokinetics in plasma of lambs was studied after co-administration of 0.2 mg kg(-1) moxidectin (subcutaneously (SC)) and 0.2 mg kg(-1) ivermectin (SC), or 10 mg kg(-1) quercetin (SC), or 10 mg kg(-1) ketoconazole (orally). Ivermectin and quercetin increased significantly the quantity of 14C moxidectin in the rat hepatocytes. Ketoconazole co-administration led to a higher concentration of moxidectin in the rat hepatocytes. In vivo, only quercetin was able to modify the pharmacokinetics of moxidectin in plasma of lambs by increasing significantly the area under the plasma concentration-time curve. This study allowed the use of a natural agent, quercetin, to improve the bioavailability of moxidectin.

Comparative distribution of ivermectin and doramectin to parasite location tissues in cattle

Pharmacokinetic studies have been used traditionally to characterize drug concentration profiles achieved in the bloodstream. However, endectocide molecules exert their persistent and broad spectrum activity against parasites localized in many different tissues. The aim of this study was to compare the distribution of ivermectin (IVM) and doramectin (DRM) to different tissues in which parasites are found following subcutaneous administration to calves. Holstein calves weighing 120-140 kg were injected in the shoulder area with commercially available formulations of IVM (Ivomec 1% MSD AGVET, NJ, USA) (Group A) or DRM (Dectomax 1%, Pfizer, NY, USA) (Group B). Two treated calves were sacrificed at 1, 4, 8, 18, 28, 38, 48 or 58 days post-treatment. Plasma, abomasal and small intestinal fluids and mucosal tissues, bile, faeces, lung and skin samples were collected, extracted, derivatized and analyzed by high performance liquid chromatography (HPLC) with fluorescence detection to determine IVM and DRM concentrations. IVM and DRM were distributed to all the tissues and fluids analyzed. Concentrations >0.1 ng/ml (ng/g) were detected between 1 and 48 days post-treatment in all the tissues and fluids investigated. At 58 days post-treatment, IVM and DRM were detected only in bile and faeces, where large concentrations were excreted. Delayed Tmax values for DRM (4 days post-administration) compared to those for IVM (1 day) were observed in the different tissues and fluids. High IVM and DRM concentrations were measured in the most important target tissues, including skin. The highest IVM and DRM concentrations were measured in abomasal mucosa and lung tissue. Enhanced availabilities of both IVM (between 45 and 244%) and DRM (20-147%) were obtained in tissues compared to plasma. There was good correlation between concentration profiles of both compounds in plasma and target tissues (mucosal tissue, skin, and lung). Drug concentrations in target tissues remained above 1 ng/g for either 18 (IVM) or 38 (DRM) days post-treatment. The characterization of tissue distribution patterns contributes to our understanding of the basis for the broad-spectrum endectocide activity of avermectin-type compounds.

Determination of moxidectin in plasma by high-performance liquid chromatography with automated solid-phase extraction and fluorescence detection.

Moxidectin is a newly developed potent anthelmintic agent with a high potency although present at very low concentration in cattle plasma. A method is described for the determination of moxidectin in plasma using high-performance liquid chromatography with fluorescence detection (excitation and emission wavelengths 383 and 447 nm, respectively). The fluorescent derivative was obtained by a dehydrative reaction with trifluoroacetic anhydride and N-methylimidazole. The method employs 1-ml plasma samples and has linear calibration graphs (r = 0.997) over the concentration range studied, i.e., 0.1-10 ng/ml. Solid-phase extraction using the Benchmate procedure was used for sample preparation. Recoveries at low concentrations (0.1-10 ng/ml) were higher than 75%. The limit of quantification was 0.1 ng/ml (C.V. 6.95%). The method is suitable for the pharmacokinetic study of moxidectin after subcutaneous administration to cows.

Eprinomectin in dairy goats : dose influence on plasma levels and excretion in milk

Abstract The plasma levels and milk excretion of eprinomectin were determined in goats following topical application at doses of 0.5 mg kg−1 and 1.0 mg kg−1. The area under the concentration–time curve (AUC) was 2 times lower for 0.5 mg kg−1 (8.24 ± 3.50 ng day−1 ml−1) than for 1.0 mg kg−1 (15.68 ± 8.84 ng day−1 ml−1), suggesting that the pharmacokinetics of eprinomectin in goats is dose independent. The bioavailability of eprinomectin in lactating compared with non-lactating goats is low. This is probably due to the physiological status of dairy animals, which present a marked decrease in body fat. Comparison of the eprinomectin concentrations in the milk and plasma demonstrated a parallel disposition of the drug with a milk-to-plasma ratio of 0.10–0.25. The amount of drug recovered in the milk was 0.3–0.5% of the total administered dose. In all cases, the maximum level of residue in milk remained below the maximum acceptable level of 30 ng ml−1 permitted in lactating cattle.

Some Pharmacokinetic Parameters of Eprinomectin in Goats following Pour-on Administration

Some pharmacokinetic parameters of eprinomectin were determined in goats following topical application at a dose rate of 0.5 mg/kg. The plasma concentration versus time data for the drug were analysed using a one-compartment model. The maximum plasma concentration of 5.60±1.01 ng/ml occurred 2.55 days after administration. The area under the concentration–time curve (AUC) was 72.31±11.15 ng day/ml and the mean residence time (MRT) was 9.42±0.43 days. Thus, the systemic availability of eprinomectin to goats was significantly lower than that for cows. The low concentration of eprinomectin in the plasma of goats suggests that the pour-on dose of 0.5 mg/kg would be less effective in this species than in cows. Further relevant information about the optimal dosage and residues in the milk of dairy goats is needed before eprinomectin should be used in this species.

Ketoconazole increases the plasma levels of ivermectin in sheep

The parasiticide ivermectin and the antifungal drug ketoconazole are drugs that interact with P-glycoprotein. We have tested the ability of ketoconazole at a clinical dose to modify the pharmacokinetics of ivermectin in sheep. Lacaune lambs were administered with a single oral dose of ivermectin alone at 0.2 mg/kg (n=5) or in combination with a daily oral dose of ketoconazole (10 mg/kg) given for 3 days before and 2 days after the ivermectin (n=5). The plasma kinetics of ivermectin and its metabolite were followed over 15 days by HPLC analysis. Co-administration of ketoconazole induced higher plasma concentrations of ivermectin, leading to a substantial increase in the overall exposure of the animals to the drug. Ketoconazole did not reduce the production of the main ivermectin metabolite but it may rather act by inhibiting P-glycoprotein, and thus increasing the absorption of ivermectin. The use of a P-gp reversing agent such as ketoconazole could be useful tool to optimize antiparasitic therapy in the face of the worldwide development of anthelmintic resistance.

Enhanced absorption of pour-on ivermectin formulation in rats by co-administration of the multidrug-resistant-reversing agent verapamil.

Abstract The effect of verapamil, a multidrug-resistance (Mdr)-reversing agent on the absorption of a pour-on formulation of ivermectin was evaluated in rats. Absorption of ivermectin was effectively enhanced (40%) by the presence of verapamil, suggesting that absorption of ivermectin involves Mdr-P-glycoprotein and that verapamil should act as a competitive inhibitor for the transport and extrusion of ivermectin by P-glycoprotein. This hypothesis is consistent with other studies describing verapamil as a blocking agent of P-glycoprotein involved in the efflux of ivermectin in a resistant strain of Haemonchus contortus.

Plasma and milk kinetic of eprinomectin and moxidectin in lactating water buffaloes (Bubalus bubalis).

The pharmacokinetics and mammary excretion of moxidectin and eprinomectin were determined in water buffaloes (Bubalus bubalis) following topical administration of 0.5mgkg(-1). Following administration of moxidectin, plasma and milk concentrations of moxidectin increased to reach maximal concentrations (C(max)) of 5.46+/-3.50 and 23.76+/-16.63ngml(-1) at T(max) of 1.20+/-0.33 and 1.87+/-0.77 days in plasma and milk, respectively. The mean residence time (MRT) were similar for plasma and milk (5.27+/-0.45 and 5.87+/-0.80 days, respectively). The AUC value was 5-fold higher in milk (109.68+/-65.01ngdayml(-1)) than in plasma (23.66+/-12.26ngdayml(-1)). The ratio of AUC milk/plasma for moxidectin was 5.04+/-2.13. The moxidectin systemic availability (expressed as plasma AUC values) obtained in buffaloes was in the same range than those reported in cattle. The faster absorption and elimination processes of moxidectin were probably due to a lower storage in fat associated with the fact that animals were in lactation. Nevertheless, due to its high excretion in milk and its high detected maximum concentration in milk which is equivalent or higher to the Maximal Residue Level value (MRL) (40ngml(-1)), its use should be prohibited in lactating buffaloes. Concerning eprinomectin, the C(max) were of 2.74+/-0.89 and 3.40+/-1.68ngml(-1) at T(max) of 1.44+/-0.20 and 1.33+/-0.0.41 days in plasma and milk, respectively. The MRT and the AUC were similar for plasma (3.17+/-0.41 days and 11.43+/-4.01ngdayml(-1)) and milk (2.70+/-0.44 days and 8.49+/-3.33ngdayml(-1)). The ratio of AUC milk/plasma for eprinomectin was 0.76+/-0.16. The AUC value is 20 times lower than that reported in dairy cattle. The very low extent of mammary excretion and the milk levels reported lower than the MRL (20ngml(-1)) supports the permitted use of eprinomectin in lactating water buffaloes.

Determination of eprinomectin in plasma by high-performance liquid chromatography with automated solid phase extraction and fluorescence detection

A method is described for the determination of eprinomectin in plasma using high-performance liquid chromatography with fluorescence detection (excitation and emission wavelengths 355 and 465 nm, respectively). The fluorescent derivative was obtained by a condensation reaction with trifluoroacetic anhydride and N-methylimidazole. The method employs 1 ml plasma samples and gives linear calibration graphs (r = 0.999) over the concentration range studied (0.5-50 ng ml-1). Solid extraction using the benchmate procedure was used for sample preparation. This method permits the determination of eprinomectin at levels as low as 0.1 ng ml-1 and is suitable for the pharmacokinetic study of eprinomectin in animals.

Identification of human and rabbit cytochromes P450 1A2 as major isoforms involved in thiabendazole 5-hydroxylation

Summary— This report characterized one of the major cytochrome P450 isozyme involved in thiabendazole metabolism. This study was undertaken by using both cultured rabbit hepatocytes treated or not with drugs known to specifically induce various cytochromes P450 isoenzymes (ie, P450 1A1/2 by β‐naphthoflavone, P450 2B4 by phenobarbital, P450 3A6 by rifampicine and P450 4A by clofibrate) and human liver (THLE‐5) and bronchial (BEAS‐2B) epithelial cells expressing or not the major constitutive human cytochromes P450 (ie, CYP1A2, 2A6, 2B6, 2C9, 2D6, 2E1 or 3A4). Only hepatocytes exposed to β‐naphthoflavone and clofibrate significantly metabolized thiabendazole to 5‐hydroxythiabendazole. Extensive biotransformation of this anthelmintic only occurred in human cells expressing CYP1A2. Moreover, experiments performed on rabbit preparations showed good correlations between thiabendazole 5‐hydroxylase activity and both ethoxyresorufin and methoxyresorufin O‐dealkylase activities. Thus, CYP1A2 is a major isoenzyme involved in thiabendazole 5‐hydroxylation.