Antithrombotic activity of LB30057, a newly synthesized direct thrombin inhibitor

Kyu-Tae Kang; Byoung-In Jung; Ok-Nam Bae; Moo-Yeol Lee; Seung-Min Chung; Joo-Young Lee; Sang-Koo Lee; In-Chul Kim; Jin-Ho Chung

doi:10.1055/s-0037-1613549

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2003; 89(01): 104-111
DOI: 10.1055/s-0037-1613549

Blood Coagulation, Fibrinolysis and Cellular Haemostasis

Schattauer GmbH

Antithrombotic activity of LB30057, a newly synthesized direct thrombin inhibitor

Authors

Kyu-Tae Kang

²Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul
Byoung-In Jung

²Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul
Ok-Nam Bae

²Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul
Moo-Yeol Lee

²Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul
Seung-Min Chung

²Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul
Joo-Young Lee

²Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul
Sang-Koo Lee

¹Biotech Research Institute, LG Chemical, Taejon, Korea
In-Chul Kim

¹Biotech Research Institute, LG Chemical, Taejon, Korea
Jin-Ho Chung

²Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul

Abstract

Summary

An amidrazonophenylalanine derivative, LB30057, inhibits the catalytic activity of thrombin potently by interaction with the active site of thrombin, and has high water solubility. In the present study, we evaluated the effect of LB30057 on the biological activities of thrombin at various tissues, and determined whether thrombin inhibition by LB30057 could reduce the incidence of occlusive thrombosis in an in vivo animal model. Treatment with LB30057 to human plasma prolonged clotting times in a concentration-dependent manner. LB30057 suppressed significantly thrombin-induced phosphatidylserine (PS) exposure in platelets, suggesting that LB30057 could inhibit blood coagulation accelerated by PS exposure. In human platelets, soluble thrombin- and clot-induced platelet aggregation was inhibited by LB30057 potently. Consistent with this finding, LB30057 showed concentration-dependent inhibitory effects on serotonin secretion and P-selectin expression induced by thrombin in platelets. In the blood vessel isolated from the guinea pig, treatment with LB30057 resulted in a concentration-dependent inhibition of thrombin-induced vascular contraction. In vivo study revealed that LB30057 following oral administration significantly increased the time to occlusion and improved carotid arterial patency using rat carotid artery thrombosis model. All these results suggest that LB30057 is a potent inhibitor of biological activities of thrombin at various target tissues and, therefore, might be developed as an anti-thrombotic agent for treatment and prevention of thrombotic diseases.

Keywords

Thrombin inhibitor - LB30057 - antithrombotic activity

Full Text

References

References
1 Tapparelli C, Metternich R, Ehrhardt C, Cook NS. Synthetic low-molecular weight thrombin inhibitors: molecular design and pharmacological profile. Trends Pharmacol Sci 1993; 14: 366-76.
2 Weiss HJ, Turitto VT, Baumgartner HR. Further evidence that glycoprotein IIb-IIIa mediates platelet spreading on subendotheli-um. Thromb Haemost 1991; 65: 202-5.
3 Stenberg PE, McEver RP, Shuman MA, Jacques YV, Bainton DF. A platelet alpha-granule membrane protein (GMP-140) is expressed on the plasma membrane after activation. J Cell Biol 1985; 101: 880-6.
4 Glusa E. Vascular effects of thrombin. Semin Thromb Hemost 1992; 18: 296-304.
5 Harker LA. Platelets and vascular thrombosis. N Engl J Med 1994; 330: 1006-7.
6 Vanhoutte PM, Houston DS. Platelets, endothelium, and vasospasm. Circulation 1985; 72: 728-34.
7 Hauptmann J, Sturzebecher J. Synthetic inhibitors of thrombin and factor Xa: from bench to bedside. Thromb Res 1999; 93: 203-41.
8 Weitz JI. Low-molecular-weight heparins. N Engl J Med 1997; 337: 688-98.
9 FitzGerald GA. The human pharmacology of thrombin inhibition. Coron Artery Dis 1996; 7: 911-8.
10 Weitz JI, Hudoba M, Massel D, Maraganore J, Hirsh J. Clot-bound thrombin is protected from inhibition by heparin- antithrombin III but is susceptible to inactivation by antithrom-bin III-independent inhibitors. J Clin Invest 1990; 86: 385-91.
11 Heras M, Chesebro JH, Penny WJ, Bailey KR, Badimon L, Fuster V. Effects of thrombin inhibition on the development of acute platelet-thrombus deposition during angioplasty in pigs. Heparin versus recombinant hirudin, a specific thrombin inhibitor. Circulation 1989; 79: 657-65.
12 Kelly AB, Marzec UM, Krupski W, Bass A, Cadroy Y, Hanson SR, Harker LA. Hirudin interruption of heparin-resistant arterial thrombus formation in baboons. Blood 1991; 77: 1006-12.
13 Markwardt F. The development of hirudin as an antithrombotic drug. Thromb Res 1994; 74: 1-23.
14 Hursting MJ, Alford KL, Becker JC, Brooks RL, Joffrion JL, Knappenberger GD, Kogan PW, Kogan TP, McKinney AA, Schwarz Jr. RP. Novastan (brand of argatroban): a small-molecule, direct thrombin inhibitor. Semin Thromb Hemost 1997; 23: 503-16.
15 Gustafsson D, Antonsson T, Bylund R, Eriksson U, Gyzander E, Nilsson I, Elg M, Mattsson C, Deinum J, Pehrsson S, Karlsson O, Nilsson A, Sorensen H. Effects of melagatran, a new ow-molecular-weight thrombin inhibitor, on thrombin and fibrinolytic enzymes. Thromb Haemost 1998; 79: 110-8.
16 Oh YS, Yun M, Hwang SY, Hong S, Shin Y, Lee K, Yoon KH, Yoo YJ, Kim DS, Lee SH, Lee YH, Park HD, Lee CH, Lee SK, Kim S. Discovery of LB30057, a benzamidrazone-based selective oral thrombin inhibitor. Bioorg Med Chem Lett 1998; 8: 631-4.
17 Holmsen H, Dangelmaier CA. Measurement of secretion of serotonin. Methods Enzymol 1989; 169: 205-10.
18 Hoffman M, Monroe DM. A cell-based model of hemostasis. Thromb Haemost 2001; 85: 958-65.
19 Dachary-Prigent J, Toti F, Satta N, Pasquet JM, Uzan A, Freyssinet JM. Physiopatho-logical significance of catalytic phospholipids in the generation of thrombin. Semin Thromb Hemost 1996; 22: 157-64.
20 Walz DA, Anderson GF, Ciaglowski RE, Aiken M, Fenton JW. Thrombin-elicited contractile responses of aortic smooth muscle. Proc Soc Exp Biol Med 1985; 180: 518-26.
21 Brass LF, Manning DR, Cichowski K, Abrams CS. Signaling through G proteins in platelets: to the integrins and beyond. Thromb Haemost 1997; 78: 581-9.
22 Fanburg BL, Lee SL. A new role for an old molecule: serotonin as a mitogen. Am J Physiol 1997; 272: L795-L806.
23 Hogg PJ, Jackson CM. Fibrin monomer protects thrombin from inactivation by heparin-antithrombin III: implications for heparin efficacy. Proc Natl Acad Sci USA 1989; 86: 3619-23.
24 Kaminski M, McDonagh J. Inhibited thrombins. Interactions with fibrinogen and fibrin. Biochem J 1987; 242: 881-7.
25 Bendayan P, Boccalon H, Dupouy D, Boneu B. Dermatan sulfate is a more potent inhibitor of clot-bound thrombin than unfractionated and low molecular weight heparins. Thromb Haemost 1994; 71: 576-80.
26 Kato Y, Kita Y, Nishio M, Hirasawa Y, Ito K, Yamanaka T, Motoyama Y, Seki J. In vitro antiplatelet profile of FR171113, a novel non-peptide thrombin receptor antagonist. Eur J Pharmacol 1999; 384: 197-202.
27 Gustafsson D, Antonsson T, Bylund R, Eriksson U, Gyzander E, Nilsson I, Elg M, Mattsson C, Deinum J, Pehrsson S, Karlsson O, Nilsson A, Sorensen H. Effects of melagatran, a new low-molecular-weight thrombin inhibitor, on thrombin and fibrinolytic enzymes. Thromb Haemost 1998; 79: 110-8.