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Importance of measuring pharmacologically active metabolites of edoxaban: development and validation of an ultra-high-performance liquid chromatography coupled with a tandem mass spectrometry method

Abstract

Although DOACs do not require regular measurements of their blood concentrations, clinical situations may require an assessment of their concentration. Among the factor Xa inhibitors, edoxaban is the only compound for which some metabolites (e.g. edoxaban-M4) are reported to be pharmacologically active. Therefore, their contribution could interfere with assays used for the estimation of edoxaban concentration. In addition, drug interactions may alter the metabolite/parent compound ratio making the sole estimation of edoxaban concentration, a poor assessment of the overall anticoagulation. To develop a validated UHPLC–MS/MS method to quantify simultaneously edoxaban and its more relevant M4-metabolite in human plasma. Electrospray ionization and chromatographic separation were optimized for the simultaneous dosage of edoxaban and edoxaban-M4. The method was validated according to regulatory guidelines for bioanalytical method validation. The total run time was 6 min. The method was validated for calibration curves, precision, accuracy, carry-over, selectivity, matrix effect and short-time stability. This method permits quantification of edoxaban and edoxaban-M4 providing complementary information about the inhibitory effect of this active metabolite in chronometric or chromogenic assays. Although patients treated with edoxaban exhibits usually low concentrations of active metabolites, the measurement of edoxaban-M4 is interesting; especially in case of drug interactions. Indeed, concomitant prescriptions of edoxaban and carbamazepine or rifampicin is frequent and may lead to disturbance of the estimations of edoxaban concentration by chromogenic anti-Xa assays. Therefore, patients are at risk of having inadequate control of anticoagulation supporting the need of measuring the most representative edoxaban metabolite concomitantly to the parent compound.

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Abbreviations

%Dev:

Percentage of relative standard deviation

CV:

Coefficient of variation

DMSO:

Dimethyl sulfoxide

DOACs:

Direct oral anticoagulants

EMA:

European Medicines Agency

ESI:

Electro spray ionization

ESI + :

Electro spray on positive ionization mode

FDA:

Food and Drug Administration

ICH:

International Council of Harmonization

IS:

Internal standard

LC:

Liquid chromatography

LLOQ:

Lower limit of quantification

ME:

Matrix effect

MRM:

Multiple reaction monitoring

MS:

Mass spectrometry

NPP:

Normal pooled plasma

PBS:

Phosphate buffer saline

PPP:

Platelet poor plasma

QCs:

Quality controls

UHPLC–MS/MS:

Ultra-high-performance liquid chromatography coupled with mass spectrometry

ULOQ:

Upper limit of quantification

VKAs:

Vitamin K antagonists

VTE:

Venous thromboembolisms

References

  1. 1.

    Siriez R et al (2019) Development of new methodologies for the chromogenic estimation of betrixaban concentrations in plasma. Int J Lab Hematol. https://doi.org/10.1111/ijlh.12963

  2. 2.

    Siriez R et al (2018) Betrixaban: impact on routine and specific coagulation assays-a practical laboratory guide. Thromb Haemost 118(7):1203–1214

  3. 3.

    Douxfils J et al (2018) Laboratory testing in patients treated with direct oral anticoagulants: A practical guide for clinicians. J Thromb Haemost 16(2):209–219

  4. 4.

    Douxfils J, Gosselin RC (2017) Laboratory assessment of direct oral anticoagulants. Semin Thromb Hemost 43(3):277–290

  5. 5.

    Douxfils J et al (2016) Edoxaban: impact on routine and specific coagulation assays A practical laboratory guide. Thromb Haemost 115(2):368–381

  6. 6.

    Douxfils J et al (2013) Impact of apixaban on routine and specific coagulation assays: a practical laboratory guide. Thromb Haemost 110(2):283–294

  7. 7.

    Douxfils J et al (2012) Assessment of the impact of rivaroxaban on coagulation assays: laboratory recommendations for the monitoring of rivaroxaban and review of the literature. Thromb Res 130(6):956–966

  8. 8.

    Douxfils J et al (2012) Impact of dabigatran on a large panel of routine or specific coagulation assays Laboratory recommendations for monitoring of dabigatran etexilate. Thromb Haemost 107(5):985–997

  9. 9.

    Skeppholm M et al (2014) On the monitoring of dabigatran treatment in "real life" patients with atrial fibrillation. Thromb Res 134(4):783–789

  10. 10.

    Parasrampuria DA, Truitt KE (2016) Pharmacokinetics and pharmacodynamics of edoxaban, a non-vitamin k antagonist oral anticoagulant that inhibits clotting factor xa. Clin Pharmacokinet 55(6):641–655

  11. 11.

    Food and Drug Administration (2014) Clinical pharmacology and biopharmaceutics review(s)

  12. 12.

    Parasrampuria DA et al (2016) Edoxaban drug–drug interactions with ketoconazole, erythromycin, and cyclosporine. Br J Clin Pharmacol 82(6):1591–1600

  13. 13.

    Food and Drug Administration (2018) Bioanalytical method validation: guidance for industry. https://www.fda.gov/downloads/drugs/guidances/ucm070107.pdf. Accessed 6 Feb 2019

  14. 14.

    He L et al (2017) Determination of edoxaban equivalent concentrations in human plasma by an automated anti-factor Xa chromogenic assay. Thromb Res 155:121–127

  15. 15.

    EMA, ICH Topic Q2 (R1) (1995) Validation of analytical procedures: text and methodology—NOTE FOR GUIDANCE ON VALIDATION OF ANALYTICAL PROCEDURES: TEXT AND METHODOLOGY (CPMP/ICH/381/95)

  16. 16.

    Rohde G (2008) Determination of rivaroxaban—a novel, oral, direct factor Xa inhibitor—in human plasma by high-performance liquid chromatography-tandem mass spectrometry. J Chromatogr B 872(1–2):43–50

  17. 17.

    Lagoutte-Renosi J et al (2018) A simple and fast HPLC–MS/MS method for simultaneous determination of direct oral anticoagulants apixaban, dabigatran, rivaroxaban in human plasma. J Chromatogr B 1100–1101:43–49

  18. 18.

    Foerster KI et al (2018) Simultaneous quantification of direct oral anticoagulants currently used in anticoagulation therapy. J Pharm Biomed Anal 148:238–244

  19. 19.

    Mendell J et al (2015) The effect of rifampin on the pharmacokinetics of edoxaban in healthy adults. Clin Drug Investig. https://doi.org/10.1007%2Fs40261-015-0298-2

  20. 20.

    Ruff CT et al (2015) Association between edoxaban dose, concentration, anti-factor Xa activity, and outcomes: an analysis of data from the randomised, double-blind ENGAGE AF-TIMI 48 trial. Lancet 385(9984):2288–2295

  21. 21.

    Verhamme P et al (2016) Dose reduction of edoxaban preserves efficacy and safety for the treatment of venous thromboembolism An analysis of the randomised, double-blind HOKUSAI VTE trial. Thromb Haemost 116(4):747–753

  22. 22.

    Weitz JI et al (2010) Randomised, parallel-group, multicentre, multinational phase 2 study comparing edoxaban, an oral factor Xa inhibitor, with warfarin for stroke prevention in patients with atrial fibrillation. Thromb Haemost 104(3):633–641

  23. 23.

    Levy JH et al (2016) When and how to use antidotes for the reversal of direct oral anticoagulants: guidance from the SSC of the ISTH. J Thromb Haemost 14(3):623–627

  24. 24.

    CBIP (2018) Centre Belge d’Information Pharmacothérapeutique - Bon usage des médicaments - Interactions des médicaments

  25. 25.

    Vazquez SR (2018) Drug-drug interactions in an era of multiple anticoagulants: a focus on clinically relevant drug interactions. Hematol Am Soc Hematol Educ Program 2018(1):339–347

  26. 26.

    Wang JZ et al (2017) Incidence and management of seizures after ischemic stroke: systematic review and meta-analysis. Neurology 89(12):1220–1228

  27. 27.

    Jabareen A et al (2018) Treatment with antiepileptic drugs in patients with stroke. A change in clinical practice may be required. J Neurol Sci 395:4–7

  28. 28.

    Galgani A et al (2018) Pharmacokinetic interactions of clinical interest between direct oral anticoagulants and antiepileptic drugs. Front Neurol 9:1067

  29. 29.

    Di Gennaro L et al (2019) Carbamazepine interaction with direct oral anticoagulants: help from the laboratory for the personalized management of oral anticoagulant therapy. J Thromb Thrombolysis. https://doi.org/10.1007/s11239-019-01866-1

  30. 30.

    Stollberger C, Finsterer J (2016) Interactions between non-vitamin K oral anticoagulants and antiepileptic drugs. Epilepsy Res 126:98–101

  31. 31.

    European Medicines Agency (2015) Lixiana: EMEA/H/C/002629/0000, 25 Apr 2015. https://www.ema.europa.eu/en/documents/assessment-report/lixiana-epar-public-assessment-report_en.pdf. Accessed 23 Oct 2019

  32. 32.

    Food and Drug Administration (2015) Savaysa: clinical pharmacology and biopharmaceutics review(s). 2015. Accessed 12 Jun 2015

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Acknowledgements

The authors would like to acknowledge Christelle Vancraeynest, Vincent Maloteau, Lionel Pochet and Jonathan Evrard for their contribution to this work. The authors would also like to thank the NAB-X biobank from the University of Namur (UNamur) for the management and storage of plasma samples.

Author information

Correspondence to Romain Siriez.

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Conflict of interest

The other authors have no conflicts of interest to disclose.

Ethical approval

Among the authors, J. Douxfils is CEO and founder of QUALIblood s.a. and reports personal fees from Portola Pharmaceuticals, Stago, Roche, Roche Diagnostics and Daiichi-Sankyo, outside the submitted work.

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Siriez, R., Alpan, L., Elasaad, K. et al. Importance of measuring pharmacologically active metabolites of edoxaban: development and validation of an ultra-high-performance liquid chromatography coupled with a tandem mass spectrometry method. J Thromb Thrombolysis (2020). https://doi.org/10.1007/s11239-019-02030-5

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Keywords

  • Edoxaban
  • Edoxaban-M4
  • Ultra-high-performance liquid chromatography/tandem mass spectrometry
  • Metabolite
  • Direct oral anticoagulant