Influence of Indomethacin on Steroid Metabolism: Endocrine Disruption and Confounding Effects in Urinary Steroid Profiling of Anti-Doping Analyses.

Anna Stoll,Michele Iannone,Maria Kristina Parr,Francesco Molaioni,Giuseppina De Gregorio,Xavier De La Torre,Francesco Botrè

doi:10.3390/metabo10110463

Abstract

Anabolic androgenic steroids (AAS) are prohibited as doping substances in sports by the World Anti-Doping Agency. Concentrations and concentration ratios of endogenous AAS (steroid profile markers) in urine samples collected from athletes are used to detect their administration. Certain (non-prohibited) drugs have been shown to influence the steroid profile and thereby sophisticate anti-doping analysis. It was shown in vitro that the non-steroidal anti-inflammatory drug (NSAID) indomethacin inhibits selected steroid-biotransformations catalyzed by the aldo-keto reductase (AKR) 1C3, which plays a key role in the endogenous steroid metabolism. Kinetic parameters for the indomethacin-mediated inhibition of the AKR1C3 catalyzed reduction in etiocholanolone were determined in vitro using two comparing methods. As NSAIDs are very frequently used (not only) by athletes, the inhibitory impact of indomethacin intake on the steroid metabolism was evaluated, and steroid profile alterations were detected in vivo (one male and one female volunteer). Significant differences between samples collected before, during or after the intake of indomethacin for selected steroid profile markers were observed. The presented results are of relevance for the interpretation of results from doping control analysis. Additionally, the administration of NSAIDs should be carefully reconsidered due to their potential as endocrine disruptors.

Highlights

Anabolic androgenic steroids (AAS) are very frequently used drugs in sports [1]
In this study we focused on the AKR1C3, which is known to oxidize 17-hydroxy steroids to their corresponding 17-oxo metabolites and vice versa
Our work demonstrates that the inhibition of AKR1C3 by indomethacin alters selected markers of the urinary steroid profile in healthy volunteers

Summary

Introduction

Anabolic androgenic steroids (AAS) are very frequently used drugs in sports [1]. Their use as doping agents is prohibited in and out of competition by the World Anti-Doping Agency (WADA; class S1 in the WADA prohibited list) [2]. To detect the misuse of those pseudo endogenous and some synthetic AAS, anti-doping laboratories monitor in a first step, concentrations and concentration ratios of selected EAAS according to the WADA technical document TD2018EAAS in urine samples collected from the athletes [3]. In case of misuse of pseudo endogenous AAS or some synthetic AAS, those steroid profile markers are altered and a confirmative method using gas chromatography combustion isotope-ratio mass-spectrometry (GC-c-IRMS) is Metabolites 2020, 10, 463; doi:10.3390/metabo10110463 www.mdpi.com/journal/metabolites. Since it has been shown that ratios of urinary steroids are stable over months and even years in adult humans [4,5,6] but show interindividual variations, the steroidal module of the Athlete

Objectives

Methods

Results

Discussion

Conclusion