Pharmacokinetic Study of Vadadustat and High-Resolution Mass Spectrometric Characterization of Its Novel Metabolites in Equines for the Purpose of Doping Control

Overview

Journal Curr Drug Metab

Publisher Bentham Science Publishers

Specialties Chemistry
Endocrinology

Date 2022 Aug 26

PMID 36017833

Authors

Hideaki Ishii

Mariko Shibuya

Kanichi Kusano

Yu Sone

Takahiro Kamiya

Ai Wakuno

Hideki Ito

Kenji Miyata

Fumio Sato

Taisuke Kuroda

Masayuki Yamada

Gary Ngai-Wa Leung

Affiliations

Soon will be listed here.

Abstract

Background: Vadadustat, a hypoxia-inducible factor prolyl hydroxylase (HIF-PHD) inhibitor, is a substance which carries a lifetime ban in both horse racing and equestrian competition. A comprehensive metabolic study of vadadustat in horses has not been previously reported.

Objective: Metabolism and elimination profiles of vadadustat in equine plasma and urine were studied for the purpose of doping control.

Methods: A nasoesophageal administration of vadadustat (3 g/day for 3 days) was conducted on three thoroughbred mares. Potential metabolites were comprehensively detected by differential analysis of full-scan mass spectral data obtained from both in vitro studies with liver homogenates and post-administration samples using liquid chromatography high-resolution mass spectrometry. The identities of metabolites were further substantiated by product ion scans. Quantification methods were developed and validated for the establishment of the excretion profiles of the total vadadustat (free and conjugates) in plasma and urine.

Results: A total of 23 in vivo and 14 in vitro metabolites (12 in common) were identified after comprehensive analysis. We found that vadadustat was mainly excreted into urine as the parent drug together with some minor conjugated metabolites. The elimination profiles of total vadadustat in post-administration plasma and urine were successfully established by using quantification methods equipped with alkaline hydrolysis for cleavage of conjugates such as methylated vadadustat, vadadustat glucuronide, and vadadustat glucoside.

Conclusion: Based on our study, for effective control of the misuse or abuse of vadadustat in horses, total vadadustat could successfully be detected for up to two weeks after administration in plasma and urine.

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References

Markham A . Vadadustat: First Approval. Drugs. 2020; 80(13):1365-1371. DOI: 10.1007/s40265-020-01383-z. View

Pergola P, Spinowitz B, Hartman C, Maroni B, Haase V . Vadadustat, a novel oral HIF stabilizer, provides effective anemia treatment in nondialysis-dependent chronic kidney disease. Kidney Int. 2016; 90(5):1115-1122. DOI: 10.1016/j.kint.2016.07.019. View

Haase V, Chertow G, Block G, Pergola P, deGoma E, Khawaja Z . Effects of vadadustat on hemoglobin concentrations in patients receiving hemodialysis previously treated with erythropoiesis-stimulating agents. Nephrol Dial Transplant. 2018; 34(1):90-99. PMC: 6322440. DOI: 10.1093/ndt/gfy055. View

Nangaku M, Kondo K, Takabe S, Ueta K, Kaneko G, Otsuka M . Vadadustat for anemia in chronic kidney disease patients on peritoneal dialysis: A phase 3 open-label study in Japan. Ther Apher Dial. 2020; 25(5):642-653. PMC: 8451920. DOI: 10.1111/1744-9987.13611. View

Eckardt K, Agarwal R, Mk Farag Y, Jardine A, Khawaja Z, Koury M . Global Phase 3 programme of vadadustat for treatment of anaemia of chronic kidney disease: rationale, study design and baseline characteristics of dialysis-dependent patients in the INNO2VATE trials. Nephrol Dial Transplant. 2020; 36(11):2039-2048. PMC: 8577631. DOI: 10.1093/ndt/gfaa204. View

Min J, Yang H, Ivan M, Gertler F, Kaelin Jr W, Pavletich N . Structure of an HIF-1alpha -pVHL complex: hydroxyproline recognition in signaling. Science. 2002; 296(5574):1886-9. DOI: 10.1126/science.1073440. View

Muchnik E, Kaplan J . HIF prolyl hydroxylase inhibitors for anemia. Expert Opin Investig Drugs. 2011; 20(5):645-56. DOI: 10.1517/13543784.2011.566861. View

Yan L, Colandrea V, Hale J . Prolyl hydroxylase domain-containing protein inhibitors as stabilizers of hypoxia-inducible factor: small molecule-based therapeutics for anemia. Expert Opin Ther Pat. 2010; 20(9):1219-45. DOI: 10.1517/13543776.2010.510836. View

Bunn H . New agents that stimulate erythropoiesis. Blood. 2006; 109(3):868-73. DOI: 10.1182/blood-2006-08-019083. View

10.

Chan M, Atasoylu O, Hodson E, Tumber A, Leung I, Chowdhury R . Potent and Selective Triazole-Based Inhibitors of the Hypoxia-Inducible Factor Prolyl-Hydroxylases with Activity in the Murine Brain. PLoS One. 2015; 10(7):e0132004. PMC: 4492579. DOI: 10.1371/journal.pone.0132004. View

11.

Beuck S, Schanzer W, Thevis M . Hypoxia-inducible factor stabilizers and other small-molecule erythropoiesis-stimulating agents in current and preventive doping analysis. Drug Test Anal. 2012; 4(11):830-45. DOI: 10.1002/dta.390. View

12.

Joharapurkar A, Pandya V, Patel V, Desai R, Jain M . Prolyl Hydroxylase Inhibitors: A Breakthrough in the Therapy of Anemia Associated with Chronic Diseases. J Med Chem. 2018; 61(16):6964-6982. DOI: 10.1021/acs.jmedchem.7b01686. View

13.

Mazzarino M, Perretti I, Stacchini C, Comunita F, de la Torre X, Botre F . UPLC-MS-Based Procedures to Detect Prolyl-Hydroxylase Inhibitors of HIF in Urine. J Anal Toxicol. 2020; 45(2):184-194. DOI: 10.1093/jat/bkaa055. View

14.

Hu C, Wang L, Chodosh L, Keith B, Simon M . Differential roles of hypoxia-inducible factor 1alpha (HIF-1alpha) and HIF-2alpha in hypoxic gene regulation. Mol Cell Biol. 2003; 23(24):9361-74. PMC: 309606. DOI: 10.1128/MCB.23.24.9361-9374.2003. View

15.

Elvert G, Kappel A, Heidenreich R, Englmeier U, Lanz S, Acker T . Cooperative interaction of hypoxia-inducible factor-2alpha (HIF-2alpha ) and Ets-1 in the transcriptional activation of vascular endothelial growth factor receptor-2 (Flk-1). J Biol Chem. 2002; 278(9):7520-30. DOI: 10.1074/jbc.M211298200. View

16.

Semenza G . A compendium of proteins that interact with HIF-1α. Exp Cell Res. 2017; 356(2):128-135. PMC: 5541399. DOI: 10.1016/j.yexcr.2017.03.041. View

17.

Chan M, Ilott N, Schodel J, Sims D, Tumber A, Lippl K . Tuning the Transcriptional Response to Hypoxia by Inhibiting Hypoxia-inducible Factor (HIF) Prolyl and Asparaginyl Hydroxylases. J Biol Chem. 2016; 291(39):20661-73. PMC: 5034057. DOI: 10.1074/jbc.M116.749291. View

18.

Thevis M, Schanzer W . Analytical approaches for the detection of emerging therapeutics and non-approved drugs in human doping controls. J Pharm Biomed Anal. 2014; 101:66-83. DOI: 10.1016/j.jpba.2014.05.020. View

19.

Beuck S, Bornatsch W, Lagojda A, Schanzer W, Thevis M . Development of liquid chromatography-tandem mass spectrometry-based analytical assays for the determination of HIF stabilizers in preventive doping research. Drug Test Anal. 2012; 3(11-12):756-70. DOI: 10.1002/dta.365. View

20.

Hansson A, Thevis M, Cox H, Miller G, Eichner D, Bondesson U . Investigation of the metabolites of the HIF stabilizer FG-4592 (roxadustat) in five different in vitro models and in a human doping control sample using high resolution mass spectrometry. J Pharm Biomed Anal. 2016; 134:228-236. DOI: 10.1016/j.jpba.2016.11.041. View