Thromb Haemost 2010; 103(04): 815-825
DOI: 10.1160/TH09-03-0176
New Technologies, Diagnostic Tools and Drugs
Schattauer GmbH

Assessment of laboratory assays to measure rivaroxaban – an oral, direct factor Xa inhibitor

Meyer Michel Samama
1   Hôtel-Dieu University Hospital, Paris, France
2   Biomnis Laboratory, Ivry-sur-Seine, France
,
Jean-Luc Martinoli
3   Stago, Asnières, France
,
Léna LeFlem
2   Biomnis Laboratory, Ivry-sur-Seine, France
,
Céline Guinet
2   Biomnis Laboratory, Ivry-sur-Seine, France
,
Geneviève Plu-Bureau
1   Hôtel-Dieu University Hospital, Paris, France
,
François Depasse
2   Biomnis Laboratory, Ivry-sur-Seine, France
,
Elisabeth Perzborn
4   Bayer Schering Pharma AG, Wuppertal, Germany
› Author Affiliations
Further Information

Publication History

Received: 19 March 2009

Accepted after major revision: 22 February 2009

Publication Date:
22 November 2017 (online)

Summary

Although there is no need for routine coagulation monitoring with rivaroxaban – an oral, direct factor Xa inhibitor – a haemostasis assay might be valuable to measure its pharmacodynamic effects. This study aimed to find assays, among those commercially available, to measure rivaroxaban pharmacodynamics. Several global conventional clotting tests, as well as clotting or chromogenic assays to measure anti-factor Xa activity, were studied. A thrombin generation test using calibrated automated thrombogram was also done. Tests were performed with the indirect factor Xa inhibitor fondaparinux for comparison. A concentration-dependent prolongation of prothrombin time (PT), dilute PT, and activated partial thromboplastin time was observed with rivaroxaban. The results varied depending on the reagents. This variability cannot be standardised with the international normalised ratio system commonly used for vitamin K antagonists. Using a standard calibration curve, PT test results can be expressed in plasma concentrations of rivaroxaban rather than PT seconds or ratio. Standard methods for HepTest and two-step prothrombinase-induced clotting time (PiCT) resulted in a paradoxical response, with low concentrations of rivaroxaban reducing clotting times. This was not observed with shorter incubation times, or when antithrombin-deficient (immunodepleted) plasma was used. The chromogenic tests found a dose-dependent relationship between anti-factor Xa activity and rivaroxaban concentration. Modified specific factor Xa chromogenic assays are being further investigated. One-step PiCT and HepTest with shortened incubation times, as well as the widely available PT assay (using a rivaroxaban calibrator) could be useful to monitor the pharmacodynamic effects of rivaroxaban accurately. Finally, all clotting and chromogenic assays showed a concentration-dependent effect induced by rivaroxaban.

 
  • References

  • 1 Reynolds MW, Fahrbach K, Hauch O. et al. Warfarin anticoagulation and outcomes in patients with atrial fibrillation: a systematic review and metaanalysis. Chest 2004; 126: 1938-1945.
  • 2 Ansell J, Hirsh J, Hylek E. et al. Pharmacology and management of the vitamin K antagonists: American College of Chest Physicians evidence-based clinical practice guidelines (8th Edition). Chest 2008; 133: 160S-198S.
  • 3 Hirsh J, Bauer KA, Donati MB. et al. Parenteral anticoagulants: American College of Chest Physicians Evidence-Based Clinical Practice Guidelines (8th Edition). Chest 2008; 133: 141S-159S.
  • 4 Walenga JM, Jeske WP, Samama MM. et al. Fondaparinux: a synthetic heparin pentasaccharide as a new antithrombotic agent. Expert Opin Investig Drugs 2002; 11: 397-407.
  • 5 Perzborn E, Strassburger J, Wilmen A. et al. In vitro and in vivo studies of the novel antithrombotic agent BAY 59–7939 ﺹ an oral, direct Factor Xa inhibitor. J Thromb Haemost 2005; 03: 514-521.
  • 6 Mann KG, Jenny RJ, Krishnaswamy S. Cofactor proteins in the assembly and expression of blood clotting enzyme complexes. Annu Rev Biochem 1988; 57: 915-956.
  • 7 Mueck W, Becka M, Kubitza D. et al. Population model of the pharmacokinetics and pharmacodynamics of rivaroxaban ﺹ an oral, direct Factor Xa inhibitor ﺹ in healthy subjects. Int J Clin Pharmacol Ther 2007; 45: 335-344.
  • 8 Mueck W, Eriksson BI, Bauer KA. et al. Population pharmacokinetics and pharmacodynamics of rivaroxaban – an oral, direct factor xa inhibitor – in patients undergoing major orthopaedic surgery. Clin Pharmacokinet 2008; 47: 203-216.
  • 9 Eriksson BI, Borris L, Dahl OE. et al. Oral, direct Factor Xa inhibition with BAY 59-7939 for the prevention of venous thromboembolism after total hip replacement. J Thromb Haemost 2006; 04: 121-128.
  • 10 Turpie AG, Fisher WD, Bauer KA. et al. BAY 59-7939: an oral, direct factor Xa inhibitor for the prevention of venous thromboembolism in patients after total knee replacement. A phase II dose-ranging study. J Thromb Haemost 2005; 03: 2479-2486.
  • 11 Gerotziafas GT, Chakroun T, Samama MM. et al. In vitro comparison of the effect of fondaparinux and enoxaparin on whole blood tissue factor-triggered thromboelastography profile. Thromb Haemost 2004; 92: 1296-1302.
  • 12 American Diagnostica Inc.. HepTest. Clotting procedures for the quantitative determination of heparin in plasma and whole blood. Available at: www.american-diagnostica.de/fileadmin/user_upload/datasheets/830.pdf
  • 13 Calatzis A, Peetz D, Haas S. et al. Prothrombinase-induced clotting time assay for determination of the anticoagulant effects of unfractionated and low-molecular-weight heparins, fondaparinux, and thrombin inhibitors. Am J Clin Pathol 2008; 130: 446-454.
  • 14 Harder S, Parisius J, Picard-Willems B. Monitoring direct FXa-inhibitors and fondaparinux by Prothrombinase-induced Clotting Time (PiCT): Relation to FXa-activity and influence of assay modifications. Thromb Res 2008; 123: 396-403.
  • 15 Thiagarajan P, Pengo V, Shapiro SS. The use of the dilute Russell viper venom time for the diagnosis of lupus anticoagulants. Blood 1986; 68: 869-874.
  • 16 Hemker HC, Giesen PL, Ramjee M. et al. The thrombogram: monitoring thrombin generation in platelet-rich plasma. Thromb Haemost 2000; 83: 589-591.
  • 17 Marlar RA, Kleiss AJ, Griffin JH. An alternative extrinsic pathway of human blood coagulation. Blood 1982; 60: 1353-1358.
  • 18 Calatzis A, Spannagl M, Gempeler-Messina P. et al. The prothrombinase-induced clotting test: A new technique for the monitoring of anticoagulants. Haemostasis 2000; 30 (Suppl. 02) 172-174.
  • 19 Graff J, Picard-Willems B, Harder S. Monitoring effects of direct FXa-inhibitors with a new one-step prothrombinase-induced clotting time (PiCT) assay: comparative in vitro investigation with heparin, enoxaparin, fondaparinux and DX 9065a. Int J Clin Pharmacol Ther 2007; 45: 237-243.
  • 20 Hemker HC, Giesen P, al Dieri R. et al. The calibrated automated thrombogram (CAT): a universal routine test for hyper- and hypocoagulability. Pathophysiol Haemost Thromb 2002; 32: 249-253.
  • 21 Kubitza D, Becka M, Voith B. et al. Safety, pharmacodynamics, and pharmacokinetics of single doses of BAY 59-7939, an oral, direct factor Xa inhibitor. Clin Pharmacol Ther 2005; 78: 412-421.
  • 22 Gerotziafas GT, Elalamy I, Depasse F. et al. In vitro inhibition of thrombin generation, after tissue factor pathway activation, by the oral, direct Factor Xa inhibitor rivaroxaban. J Thromb Haemost 2007; 05: 886-888.
  • 23 Lormeau JC, Herault JP. The effect of the synthetic pentasaccharide SR 90107/ORG 31540 on thrombin generation ex vivo is uniquely due to ATIII-mediated neutralization of factor Xa. Thromb Haemost 1995; 74: 1474-1477.
  • 24 Samama MM, Le Flem L, Guinet C. et al. Three different patterns of calibrated automated thrombogram obtained with six different anticoagulants. J Thromb Haemost 2007; 05: 2554-2556.
  • 25 Dyke CK, Becker RC, Kleiman NS. et al. First experience with direct Factor Xa inhibition in patients with stable coronary disease: a pharmacokinetic and pharmacodynamic evaluation. Circulation 2002; 105: 2385-2391.
  • 26 Samama MM, Gerotziafas GT, Elalamy I. et al. Biochemistry and clinical pharmacology of new anticoagulant agents. Pathophysiol Haemost Thromb 2002; 32: 218-224.
  • 27 Samama MM, Le Flem L, Guinet C. et al. Suitability of chromogenic anti-FXa methods to measure rivaroxaban in human plasma. Boston ISTH 2009, poster ref. PP-WE-199. 2009.
  • 28 Perzborn E, Harwardt M. Inhibition of thrombin generation in human plasma by rivaroxaban, an oral, direct FXa inhibitor. Boston ISTH 2009, poster ref. PPMO-184. 2009.