A novel thromboxane receptor antagonist, nstpbp5185, inhibits platelet aggregation and thrombus formation in animal models

Summary

A novel benzimidazole derivative, nstpbp5185, was discovered through in vitro and in vivo evaluations for antiplatelet activity. Thro-maboxane receptor (TP) is important in vascular physiology, haemostasis and pathophysiological thrombosis. Nstpbp5185 concentration-dependently inhibited human platelet aggregation caused by collagen, arachidonic acid and U46619. Nstpbp5185 caused a right-shift of the concentration-response curve of U46619 and competitively inhibited the binding of ³H-SQ-29548 to TP receptor expressed on HEK-293 cells, with an IC₅₀ of 0.1 μM, indicating that nstpbp5185 is a TP antagonist. In murine thrombosis models, nstpbp5185 significantly prolonged the latent period in triggering platelet plug formation in mesenteric and FeCl₃-induced thrombi formation, and increased the survival rate in pulmonary embolism model with less bleeding than aspirin. This study suggests nstpbp5185, an orally selective antithrombotic agent, acting through blockade of TXA₂ receptor, may be efficacious for prevention or treatment of pathologic thrombosis.

Supplementary Material to this article is available online at www.thrombosis-online.com.

Keywords

Benzimidazole - mouse thrombotic model - thromboxane receptor antagonist

• References

• 1 Bhatt DL, Topol EJ.. Scientific and therapeutic advances in antiplatelet therapy. Nat Rev Drug Discov 2003; 2: 15-28.
• 2 Eisert WG.. How to get from antiplatelet to antithrombotic treatment. Am J Ther 2001; 8: 443-449.
• 3 Jackson SP, Schoenwaelder SM.. Antiplatelet therapy: in search of the 'magic bullet. Nat Rev Drug Discov 2003; 2: 775-789.
• 4 Hamberg M. et al. Thromboxanes: a new group of biologically active compounds derived from prostaglandin endoperoxides. Proc Natl Acad Sci USA 1975; 72: 2994-2998.
• 5 Moncada S, Vane JR.. Pharmacology and endogenous roles of prostaglandin endoperoxides, thromboxane A2, and prostacyclin. Pharmacol Rev 1978; 30: 293-331.
• 6 de Leval X. et al. New developments on thromboxane and prostacyclin modulators part II: prostacyclin modulators. Curr Med Chem 2004; 11: 1243-1252.
• 7 Dogne JM. et al. New developments on thromboxane and prostacyclin modulators part I: thromboxane modulators. Curr Med Chem 2004; 11: 1223-1241.
• 8 Bauer J. et al. Pathophysiology of isoprostanes in the cardiovascular system: implications of isoprostane-mediated thromboxane A2 receptor activation. Br J Pharmacol 2014; 171: 3115-3131.
• 9 Collaboration AT.. Collaborative meta-analysis of randomised trials of antiplatelet therapy for prevention of death, myocardial infarction, and stroke in high risk patients. Br Med J 2002; 324: 71-86.
• 10 Tantry US. et al. Resistance to antiplatelet drugs: current status and future research. Expert Opin Pharmacother 2005; 6: 2027-2045.
• 11 Awtry EH, Loscalzo J.. Aspirin. Circulation 2000; 101: 1206-1218.
• 12 Angiolillo DJ. et al. Influence of aspirin resistance on platelet function profiles in patients on long-term aspirin and clopidogrel after percutaneous coronary intervention. Am J Cardiol 2006; 97: 38-43.
• 13 Patrono C.. Aspirin resistance: definition, mechanisms and clinical read-outs. J Thromb Haemost 2003; 1: 1710-1713.
• 14 Davi G, Patrono C.. Platelet activation and atherothrombosis. N Engl J Med 2007; 357: 2482-2494.
• 15 Davi G. et al. Thromboxane receptors antagonists and/or synthase inhibitors. Handb Exp Pharmacol 2012; 261-286.
• 16 Franchini M, Mannucci PM.. New antiplatelet agents: why they are needed. Eur J Intern Med 2009; 20: 733-738.
• 17 Meade EA. et al. Differential inhibition of prostaglandin endoperoxide synthase (cyclooxygenase) isozymes by aspirin and other non-steroidal anti-inflammatory drugs. J Biol Chem 1993; 268: 6610-6614.
• 18 Bots ML. et al. Thromboxane prostaglandin receptor antagonist and carotid atherosclerosis progression in patients with cerebrovascular disease of ischaemic origin: a randomized controlled trial. Stroke 2014; 45: 2348-2353.
• 19 Chamorro A.. TP receptor antagonism: a new concept in atherothrombosis and stroke prevention. Cerebrovasc Dis 2009; 27 (Suppl. 03) 20-27.
• 20 Smirnov AV. et al. Antithrombotic activity of a new benzimidazole derivative in the thrombosis model in mice. Bull Exp Biol Med 2014; 157: 580-582.
• 21 Mustard JF. et al. Preparation of suspensions of washed platelets from humans. Br J Haematol 1972; 22: 193-204.
• 22 Born GV, Cross MJ.. The Aggregation of Blood Platelets. J Physiol 1963; 168: 178-195.
• 23 Chang MC. et al. Antithrombotic effect of crotalin, a platelet membrane glycoprotein Ib antagonist from venom of Crotalus atrox. Blood 1998; 91: 1582-1589.
• 24 Naik MU. et al. JAM-A protects from thrombosis by suppressing integrin alphaIIbbeta3-dependent outside-in signalling in platelets. Blood 2012; 119: 3352-3360.
• 25 Dejana E, Villa S, de Gaetano G.. Bleeding time in rats: a comparison of different experimental conditions. Thromb Haemost 1982; 48: 108-111.
• 26 Vroling B. et al. GPCRDB: information system for G protein-coupled receptors. Nucleic acids Res 2011; 39 (Database issue) D309-319.
• 27 Merten M, Thiagarajan P.. P-selectin in arterial thrombosis. Z Kardiol 2004; 93: 855-863.
• 28 Atkinson BT. et al. Activation of GPVI by collagen is regulated by alpha2beta1 and secondary mediators. J Thromb Haemost 2003; 1: 1278-1287.
• 29 Nieswandt B, Watson SP.. Platelet-collagen interaction: is GPVI the central receptor?. Blood 2003; 102: 449-461.
• 30 Gallet C. et al. Tyrosine phosphorylation of cortactin associated with Syk accompanies thromboxane analogue-induced platelet shape change. J Biol Chem 1999; 274: 23610-23616.
• 31 Li Z. et al. Signalling during platelet adhesion and activation. Arterioscl Thromb Vasc Biol 2010; 30: 2341-2349.
• 32 Miggin SM, Kinsella BT.. Regulation of extracellular signal-regulated kinase cascades by alpha- and beta-isoforms of the human thromboxane A(2) receptor. Mol Pharmacol 2002; 61: 817-831.
• 33 Eikelboom JW. et al. Aspirin-resistant thromboxane biosynthesis and the risk of myocardial infarction, stroke, or cardiovascular death in patients at high risk for cardiovascular events. Circulation 2002; 105: 1650-1655.
• 34 Mehta JL. et al. Increased prostacyclin and thromboxane A2 biosynthesis in atherosclerosis. Proc Natl Acad Sci USA 1988; 85: 4511-4515.
• 35 Saldeen TG. et al. Increased production of thromboxane A2 by coronary arteries after thrombolysis. Am Heart J 1993; 125: 277-284.
• 36 Needleman P. et al. Thromboxanes: selective biosynthesis and distinct biological properties. Science 1976; 193: 163-165.
• 37 Needleman P. et al. Identification of an enzyme in platelet microsomes which generates thromboxane A2 from prostaglandin endoperoxides. Nature 1976; 261: 558-560.
• 38 Ellis EF. et al. Coronary arterial smooth muscle contraction by a substance released from platelets: evidence that it is thromboxane A2. Science 1976; 193: 1135-1137.
• 39 Pawlowski NA. et al. Arachidonic acid metabolism by human monocytes. Studies with platelet-depleted cultures. J Exp Med 1983; 158: 393-412.
• 40 Coleman RA. et al. International Union of Pharmacology classification of prostanoid receptors: properties, distribution, and structure of the receptors and their subtypes. Pharmacol Rev 1994; 46: 205-229.
• 41 Halushka PV.. Thromboxane A(2) receptors: where have you gone?. Prostaglandins Other Lipid Mediat 2000; 60: 175-189.
• 42 Cheng Y. et al. Role of prostacyclin in the cardiovascular response to thromboxane A2. Science 2002; 296: 539-541.
• 43 Mehta J. et al. Platelet aggregation studies in coronary artery disease. Past 4. Effect of aspirin. Atherosclerosis 1978; 31: 169-175.
• 44 Capodanno D. et al. Antiplatelet therapy: new pharmacological agents and changing paradigms. J Thromb Haemost 2013; 11 (Suppl. 01) 316-329.
• 45 Gijon MA, Leslie CC.. Regulation of arachidonic acid release and cytosolic phospholipase A2 activation. J Leukoc Biol 1999; 65: 330-336.
• 46 Alessandrini P. et al. Physiologic variables affecting thromboxane B2 production in human whole blood. Thromb Res 1985; 37: 1-8.
• 47 Khasawneh FT. et al. Characterisation of isoprostane signalling: evidence for a unique coordination profile of 8-iso-PGF(2alpha) with the thromboxane A(2) receptor, and activation of a separate cAMP-dependent inhibitory pathway in human platelets. Biochem Pharmacol 2008; 75: 2301-2315.
• 48 Patrono C, FitzGerald GA.. Isoprostanes: potential markers of oxidant stress in atherothrombotic disease. Arterioscler Thromb Vasc Biol 1997; 17: 2309-2315.
• 49 Ting HJ, Khasawneh FT.. Platelet function and Isoprostane biology. Should isoprostanes be the newest member of the orphan-ligand family?. J Biomed Sci 2010; 17: 24.
• 50 Dogne JM. et al. Pharmacological characterisation of N-tert-butyl-N’-[2-(4’-methylphenylamino)-5-nitrobenzenesulfonyl]urea (BM-573), a novel thromboxane A2 receptor antagonist and thromboxane synthase inhibitor in a rat model of arterial thrombosis and its effects on bleeding time. J Pharmacol Exp Ther 2004; 309: 498-505.
• 51 Bousser MG. et al. Terutroban versus aspirin in patients with cerebral ischaemic events (PERFORM): a randomised, double-blind, parallel-group trial. Lancet 2011; 377: 2013-2022.