BMS-593214, an active site-directed factor VIIa inhibitor: Enzyme kinetics, antithrombotic and antihaemostatic studies

Pancras C. Wong; Joseph M. Luettgen; Alan R. Rendina; Charles A. Kettner; Baomin Xin; Robert M. Knabb; Ruth R. Wexler; E. Scott Priestley

doi:10.1160/TH10-01-0025

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2010; 104(02): 261-269
DOI: 10.1160/TH10-01-0025

Blood Coagulation, Fibrinolysis and Cellular Haemostasis

Schattauer GmbH

BMS-593214, an active site-directed factor VIIa inhibitor: Enzyme kinetics, antithrombotic and antihaemostatic studies

Pancras C. Wong

¹Thrombosis Research, Bristol-Myers Squibb Company, Pennington, New Jersey, USA

,

Joseph M. Luettgen

¹Thrombosis Research, Bristol-Myers Squibb Company, Pennington, New Jersey, USA

,

Alan R. Rendina

¹Thrombosis Research, Bristol-Myers Squibb Company, Pennington, New Jersey, USA

,

Charles A. Kettner

²Discovery Chemistry, Bristol-Myers Squibb Company, Pennington, New Jersey, USA

,

Baomin Xin

³Pharmaceutical Candidate Optimization, Bristol-Myers Squibb Company, Pennington, New Jersey, USA

,

Robert M. Knabb

¹Thrombosis Research, Bristol-Myers Squibb Company, Pennington, New Jersey, USA

,

Ruth R. Wexler

²Discovery Chemistry, Bristol-Myers Squibb Company, Pennington, New Jersey, USA

,

E. Scott Priestley

²Discovery Chemistry, Bristol-Myers Squibb Company, Pennington, New Jersey, USA

› Author Affiliations

Abstract

PDF Download

Summary

Factor (F) VIIa in association with tissue factor (TF) is the primary in vivo initiator of blood coagulation and activates FX and FIX to generate thrombin, which plays a key role in the pathogenesis of thrombosis. We evaluated the enzyme kinetics, antithrombotic and antihaemostatic properties of BMS-593214, an active-site, direct FVIIa inhibitor. Studies were conducted in enzymatic assays, and in anesthetised rabbit models of electrically-induced carotid arterial thrombosis (AT), thread-induced vena cava venous thrombosis (VT) and cuticle bleeding time (BT). Antithrombotic efficacy of BMS-593214 given intravenously was evaluated for both the prevention and treatment of AT and VT. BMS-593214 displayed direct, competitive inhibition of human FVIIa in the hydrolysis of a tripeptide substrate with K_i of 5 nM. However, it acted as a noncompetitive inhibitor of the activation of the physiological substrate FX by TF/VIIa with K_i of 9.3 nM. BMS-593214 showed selectivity for FVIIa and exhibited species differences in TF-FVIIa-dependent anticoagulation with similar potency in human and rabbit plasma. BMS-593214 was efficacious in the prevention and treatment models of AT and VT with ED₅₀ values of 1.1 to 3.1 mg/kg. Furthermore, BMS-593214 exhibited a wide therapeutic window with respect to BT. These results suggest that inhibition of FVIIa with small-molecule active-site inhibitors represents a promising antithrombotic approach for the development of new therapies for the prevention and treatment of AT and VT.

Keywords

Anticoagulant - animal models - haemostasis - thrombosis - tissue factor / factor VIIa

PDF (379 kb)

References

References
1 Weitz I J. New oral antiocagulants in development. Thromb Haemost 2010; 103: 62-70.
2 Phillips KW, Ansell J. The clinical implications of new oral anticoagulants: will the potential advantages be achieved?. Thromb Haemost 2010; 103: 34-39.
3 Mackman N, Tilley RE, Key NS. Role of the extrinsic pathway of blood coagulation in hemostasis and thrombosis. Arterioscler Thromb Vasc Biol 2007; 27: 1687-1693.
4 Ott I. Tissue factor inhibition: another approach reducing thrombosis after vascular injury. Thromb Haemost 2010; 103: 7-8.
5 Lee AY, Vlasuk GP. Recombinant nematode anticoagulant protein c2 and other inhibitors targeting blood coagulation factor VIIa/tissue factor. J Intern Med 2003; 254: 313-321.
6 Steffel J, Lüscher TF, Tanner FC. Tissue factor in cardiovascular diseases: molecular mechanisms and clinical implications. Circulation 2006; 113: 722-731.
7 Shirk RA, Vlasuk GP. Inhibitors of Factor VIIa/tissue factor. Arterioscler Thromb Vasc Biol 2007; 27: 1895-1900.
8 Harker LA, Hanson SR, Wilcox JN. et al. Antithrombotic and antilesion benefits without hemorrhagic risks by inhibiting tissue factor pathway. Haemostasis 1996; 26 (Suppl. 01) 76-82.
9 Kelley RF, Refino CJ, O'Connell MP. et al. A soluble tissue factor mutant is a selective anticoagulant and antithrombotic agent. Blood 1997; 89: 3219-3227.
10 Himber J, Kirchhofer D, Riederer M. et al. Dissociation of antithrombotic effect and bleeding time prolongation in rabbits by inhibiting tissue factor function. Thromb Haemost 1997; 78: 1142-1149.
11 Jiao J, Kelly AB, Marzec UM. et al. Inhibition of acute vascular thrombosis in chimpanzees by an anti-human tissue factor antibody targeting the factor X binding site. Thromb Haemost 2010; 103: 224-233.
12 Suleymanov OD, Szalony JA, Salyers AK. et al. Pharmacological interruption of acute thrombus formation with minimal hemorrhagic complications by a small molecule tissue factor/factor VIIa inhibitor: comparison to factor Xa and thrombin inhibition in a nonhuman primate thrombosis model. J Pharmacol Exp Ther 2003; 306: 1115-1121.
13 Szalony JA, Suleymanov OD, Salyers AK. et al. Administration of a small molecule tissue factor/factor VIIa inhibitor in a non-human primate thrombosis model of venous thrombosis: effects on thrombus formation and bleeding time. Thromb Res 2003; 112: 167-174.
14 Olivero AG, Eigenbrot C, Goldsmith R. et al. A selective, slow binding inhibitor of factor VIIa binds to a nonstandard active site conformation and attenuates thrombus formation in vivo. J Biol Chem 2005; 280: 9160-9169.
15 Young WB, Mordenti J, Torkelson S. et al. Factor VIIa inhibitors: chemical optimization, preclinical pharmacokinetics, pharmacodynamics, and efficacy in an arterial baboon thrombosis model. Bioorg Med Chem Lett 2006; 16: 2037-2041.
16 Groebke Zbinden K, Banner DW. et al. Dose-dependent antithrombotic activity of an orally active tissue factor/factor VIIa inhibitor without concomitant enhancement of bleeding propensity. Bioorg Med Chem 2006; 14: 5357-5369.
17 Zhou J, Robinson L, Gubernator NM. et al. Tetrahydroquinoline derivatives as antithrombotic agents. 2004; US Patent number: US6825208 B2.
18 De Lucca I, Zhou J, Saiah E. et al. A selective FVIIa inhibitor with efficacy in an arterial thrombosis model. Abstracts of Papers, 230th ACS National Meeting, Washington, DC, United States, Aug. 28-Sept. 1. 2005
19 Wong PC, Crain EJ, Xin B. et al. Apixaban, an oral, direct and highly selective factor Xa inhibitor: in vitro, antithrombotic and antihemostatic studies. J Thromb Haemost 2008; 06: 820-829.
20 Krishnaswamy S, Betz A. Exosites determine the macromolecular substrate recognition by prothrombinase. Biochemistry 1997; 36: 12080-12086.
21 Mousa SA, Wityak J. Orally active isoxazoline GPIIb/IIIa antagonists. Cardiovasc Drug Rev 1998; 16: 48-61.
22 Quan ML, Lam PY, Han Q. et al. Discovery of 1-(3'-aminobenzisoxazol-5'-yl)-3-trifluoromethyl-N-[2-fluoro-4– [(2'-dimethylaminomethyl)imidazol-1-yl]phenyl]-1H-pyrazole-5-carboxyamide hydrochloride (razaxaban), a highly potent, selective, and orally bioavailable factor Xa inhibitor. J Med Chem 2005; 48: 1729-1744.
23 Wong PC, Crain EJ, Watson CA. et al. Favorable therapeutic index of the direct factor Xa inhibitors, apixaban and rivaroxaban, compared with the thrombin inhibitor dabigatran in rabbits. J Thromb Haemost 2009; 07: 1313-1320.
24 Baugh RJ, Dickinson CD, Ruf W. et al. Exosite interactions determine the affinity of factor X for the extrinsic Xase complex. J Biol Chem 2000; 275: 28826-28833.
25 Rezaie AR. DX-9065a inhibition of factor Xa and the prothrombinase complex: mechanism of inhibition and comparison with therapeutic heparins. Thromb Haemost 2003; 89: 112-121.
26 Bajaj SP, Joist JH. New insights into how blood clots: implications for the use of APTT and PT as coagulation screening tests and in monitoring of anticoagulant therapy. Semin Thromb Hemost 1999; 25: 407-418.
27 Wong PC, Crain EJ, Watson CA. et al. Nonpeptide factor Xa inhibitors III: Effects of DPC423, an orally-active pyrazole antithrombotic agent, on arterial thrombosis in rabbits. J Pharmacol Exp Ther 2002; 303: 993-1000.
28 Himber J, Wohlgensinger C, Roux S. et al. A. Inhibition of tissue factor limits the growth of venous thrombus in the rabbit. J Thromb Haemost 2003; 01: 889-895.
29 Morrissey JH. Tissue factor: in at the start… and the finish?. J Thromb Haemost 2003; 01: 878-880.
30 Wong PC, Jiang X. Apixaban, a direct factor Xa inhibitor, inhibits tissue factor-induced human platelet aggregation in vitro: Comparison with direct inhibitors of factor VIIA, XIa and thrombin. Thromb Haemost 2010; 104: 302-310.
31 Wong PC, Crain EJ, Watson CA. et al. Clopidogrel versus prasugrel in rabbits. Effects on thrombosis, haemostasis, platelet function and response variability. Thromb Haemost 2009; 101: 108-115.
32 Mackman N. Tissue-specific hemostasis in mice. Arterioscler Thromb Vasc Biol 2005; 25: 2273-2281.