Functional phenotyping of the CYP2D6 probe drug codeine in the horse

Abstract

Background: In humans, the drug metabolizing enzyme CYP2D6 is highly polymorphic resulting in substantial differences in the metabolism of drugs including anti-arrhythmics, neuroleptics, and opioids. The objective of this study was to phenotype a population of 100 horses from five different breeds and assess differences in the metabolic activity of the equine CYP2D6 homolog using codeine as a probe drug. Administration of a probe drug is a common method used for patient phenotyping in human medicine, whereby the ratio of parent drug to metabolite (metabolic ratio, MR) can be used to compare relative enzyme function between individuals. A single oral dose of codeine (0.6 mg/kg) was administered and plasma concentrations of codeine and its metabolites were determined using liquid chromatography mass spectrometry. The MR of codeine O-demethylation [(codeine)/(morphine + morphine-3-glucuronide + morphine-6-glucuronide)] was determined using the area under the plasma concentration-time curve extrapolated from time zero to infinity (AUC_0-∞) for each analyte and used to group horses into predicted phenotypes (high-, moderate-, and low-MR).

Results: The MR of codeine O-demethylation ranged from 0.002 to 0.147 (median 0.018) among all horses. No significant difference in MR was observed between breeds, age, or sex. Of the 100 horses, 11 were classified as high-MR, 72 moderate-MR, and 17 low-MR. Codeine AUC_0-∞ and O-demethylation MR were significantly different (p < 0.05) between all three groups. The mean ± SD MR was 0.089 ± 0.027, 0.022 ± 0.011, and 0.0095 ± 0.001 for high-, moderate-, and low-MR groups, respectively. The AUC for the morphine metabolites morphine-3-glucuronide and morphine-6-glucuronide were significantly different between high-and low-MR groups (p < 0.004 and p < 0.006).

Conclusions: The MR calculated from plasma following codeine administration allowed for classification of horses into metabolic phenotypes within a large population. The range of codeine metabolism observed among horses suggests the presence of genetic polymorphisms in CYP2D82 of which codeine is a known substrate. Additional studies including CYP2D82 genotyping of high- and low-MR individuals are necessary to determine the presence of CYP2D polymorphisms and their functional implications with respect to the metabolism of therapeutics.

Keywords: CYP450; Codeine; Horse; Metabolic ratio; Phenotyping.

Fig. 1

Major codeine metabolites in the human and enzymes responsible for metabolite formation

Fig. 2

Normal probability plot of logarithmic metabolic ratio [log (AUC_0-∞ codeine) / (AUC_0-∞ morphine + AUC_0-∞ M3G + AUC_0-∞ M6G)] in 100 horses after a single oral administration of codeine (0.6 mg/kg)

Fig. 3

Frequency distribution histogram of log metabolic ratio [log (AUC_0-∞ codeine) / (AUC_0-∞ morphine + AUC_0-∞ M3G + AUC_0-∞ M6G)] in 100 horses after a single oral administration of codeine (0.6 mg/kg) grouped by predicted phenotype. Dashed vertical lines denote anti-modes

Fig. 4

Mean plasma-concentration time curves for codeine (a), morphine (b), C6G (c), M3G (d), norcodeine (e), and M6G (f) based on predicted phenotype in horses following a single oral dose of codeine (0.6 mg/kg)