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Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)

The use of caffeine as a metabolic probe for human drug metabolizing enzymes.

1. Caffeine (CA) is metabolized extensively and at least 17 metabolites arising from primary and secondary biotransformation pathways are found in urine following CA ingestion. The enzymes responsible for the formation of most of the metabolites derived from CA have been identified. 2. Given the near ubiquitous consumption of CA, this compound potentially constitutes a useful substrate probe for assessment of certain xenobiotic metabolizing enzyme activities in vivo. Indeed, various ratios of CA metabolites excreted in urine (urinary metabolic ratios; MRs) are now utilized widely for the population screening of enzyme activities. 3. Excretion of the acetylated secondary metabolite 5-actylamino-6-formylamino-3-methyluracil (AFMU) is dependent on the activity of the polymorphic N-acetyltransferase (NAT2), and certain MRs incorporating AFMU may be used for NAT2 phenotyping. 4. The conversion of 1-methylxanthine (1-MX), another secondary metabolite of CA, to 1-methyluric acid (1-MU) is catalyzed by xanthine oxidase ( XO), and the urinary 1-MU to 1MX ratio reflects XO activity. 5. N3-demethylation to form paraxanthine (PX), a reaction mediated by cytochrome P4501A2 (CYP1A2), is the dominant primary metabolic pathway of CA. CA N3-demethylation activity may be used as a measure of human hepatic CYP1A2 in vitro. 6. Plasma CA clearance is considered to reflect CYP1A2 activity in vivo. Although a number of MRs are based on the excretion of PX metabolites (PX derived from CA is employed for the assessment of CYP1A2 activity in vivo), factors other than enzyme activity may affect these ratios.[1]


  1. The use of caffeine as a metabolic probe for human drug metabolizing enzymes. Miners, J.O., Birkett, D.J. Gen. Pharmacol. (1996) [Pubmed]
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