Non-steroidal anti-inflammatory drugs do not influence the urinary testosterone/epitestosterone glucuronide ratio

Abstract

The UDP Glucuronosyl Transferase (UGT) enzymes are important in the pharmacokinetics, and conjugation, of a variety of drugs including non-steroidal anti-inflammatory drugs (NSAIDs) as well as anabolic androgenic steroids (AAS). Testosterone glucuronidation capacity is strongly associated with a deletion polymorphism in the UGT2B17 gene. As the use of high doses of NSAIDs has been observed in athletes there is a risk for a drug-drug interaction that may influence the doping tests for AAS. In vitro studies show inhibitory potential on UGT2B7, 2B15, and 2B17 enzymes by NSAIDs. The aim of this study was to investigate if concomitant use of NSAIDs and a single dose of testosterone enanthate would affect the excretion rate of testosterone and epitestosterone glucuronide (TG and EG) as well as the T/E ratio, thereby affecting the outcome of the testosterone doping test. The study was designed as an open, randomized, cross-over study with subjects being their own control. The 23 male healthy volunteers, with either two, one or no allele (ins/ins, ins/del, or del/del) of the UGT2B17 gene, received the maximum recommended dose of NSAID (Ibuprofen or Diclofenac) for 6 days. On day three, 500 mg of testosterone enanthate was administered. Spot urine samples were collected for 17 days. After a wash-out period of 4 months the volunteers received 500 mg testosterone enanthate only, with subsequent spot urine collection for 14 days. The glucuronides of testosterone and epitestosterone were quantified. NSAIDs did not affect the excretion of TG or EG before the administration of testosterone. The concomitant use of NSAIDs and testosterone slightly increased the TG excretion while the EG excretion was less suppressed compared to testosterone use only. The effects of the NSAIDs on the TG and EG excretion did not differ between the UGT2B17 genotype groups. In conclusion, the outcome of testosterone doping tests does not seem to be affected by the use of NSAIDs.

Keywords: NSAID; T/E ratio; UGT2B17; diclofenac; epitestosterone; ibuprofen; testosterone.

Figure 1

Illustration of the open, randomized, cross-over study with subjects being their own control. The wash-out period was at least 4 months.

Figure 2

Average urinary testosterone/epitestosterone ratios (A) and urinary testosterone glucuronide excretion (ng/μmol creatinine) (B) during 14 days in UGT2B17 del/del (top), ins/del (middle) and ins/ins (bottom) genotype groups of healthy volunteer males. The subjects were followed during two administration cycles with at least 4 months wash-out between the cycles. During one cycle NSAID was administered for 6 days (day-3 to day-2). In both cycles an intramuscular dose of 500 mg testosterone enanthate was administered on day-0. Asterisks denote statistically significant differences between the groups (*p < 0.05; **p < 0.01). Vertical bars denote standard deviations.

Figure 3

Average urinary epitestosterone glucuronide excretion (ng/μmol creatinine) during 14 days in healthy volunteer males. The subjects were followed during two administration cycles with at least 4 months wash-out between the cycles. During one cycle NSAID was administered for 6 days (day-3 to day-2). In both cycles an intramuscular dose of 500 mg testosterone enanthate was administered on day-0. Asterisks denote statistically significant differences between the groups (*p < 0.05; **p < 0.01) Vertical bars denote standard deviations.