Download PDF
Thromb Haemost 1998; 79(02): 423-430
DOI: 10.1055/s-0037-1615002
Letters to the Editor
Schattauer GmbH

Anti-thrombotic Effects of CX-397, a Recombinant Hirudin Analog, in a Canine Model of Coronary Artery Thrombosis

Tadashi Ohyama
1   Pharmacology, Development Research Laboratories, Kaken Pharmaceutical Co., Ltd., Kyoto, Japan
,
Toshimitsu Hori
1   Pharmacology, Development Research Laboratories, Kaken Pharmaceutical Co., Ltd., Kyoto, Japan
,
Mie Moriike
1   Pharmacology, Development Research Laboratories, Kaken Pharmaceutical Co., Ltd., Kyoto, Japan
,
Taiji Asano
1   Pharmacology, Development Research Laboratories, Kaken Pharmaceutical Co., Ltd., Kyoto, Japan
,
Hideya Hayashi
2   Pharmaceuticals and Biotechnology Laboratory, Japan Energy Corporation, Saitama, Japan
,
Kazunori Iwade
3   Department of Cardiology, The Heart Institute of Japan, Tokyo Women’s Medical College, Tokyo, Japan
,
Saichi Hosoda
3   Department of Cardiology, The Heart Institute of Japan, Tokyo Women’s Medical College, Tokyo, Japan
› Author Affiliations
Further Information

Publication History

Received 11 February 1997

Accepted after resubmission 10 September 1997

Publication Date:
08 December 2017 (online)

Also available at
    Permissions and Reprints

Summary

CX-397, a recombinant hirudin analog, is a potent and specific inhibitor of human α-thrombin. We conducted a comparative study of CX-397 and heparin in a canine model of left circumflex (LCX) coronary artery thrombosis to evaluate the anti-thrombotic efficacy of CX-397. Administration of drugs (i. v.; bolus + infusion) was commenced 10 min prior to the initiation of LCX coronary artery electrolytic injury (100 μA for 300 min). All saline-treated control animals (7/7) developed thrombotic occlusion during the experimental period, leaving a residual thrombus mass of 15.4 ± 3.8 mg. Treatment with CX-397 at three incremental dose levels reduced the incidence of occlusion (4/7, 2/5 and 0/7) and decreased thrombus weight (12.6 ± 2.5 mg, 6.3 ± 3.0 mg and 2.1 ± 1.3 mg, respectively) in a dose-dependent manner. At the in termediate dose (15,000 ATU/kg + 15,000 ATU/kg/h) or higher, CX-397 showed significant anti-thrombotic effects (p <0.05 and p <0.01) and suppressed increases in thrombin-antithrombin III complex (TAT) levels (p <0.01 and p <0.001). In the heparin (80 U/kg + 60 U/kg/h)-treated group, the incidence of occlusion (5/7) and thrombus weight (14.1 ± 6.2 mg) did not differ significantly from the control group. Plas ma TAT levels in the heparin group decreased compared with the control group (p <0.01), but was less potent than the intermediate dose CX-397 (p <0.01). The anti-coagulation (activated partial thromboplastin time and activated clotting time) and template bleeding time prolongation effects of heparin were more potent than those of the intermediate dose CX-397 which showed significant anti-thrombotic effects. In conclusion, CX-397 dose-dependently suppressed thrombus formation by inhibition of thrombin activity in a canine coronary artery injury model. The anti-thrombotic efficacy of CX-397 was more potent than that of heparin at equivalent anti-coagulation dosage.

 

  • References

  • 1 Fuster V, Steele PM, Chesebro JH. Role of platelets and thrombosis in coronary atherosclerotic disease and sudden death. J Am Coll Cardiol 1985; 5: 175B-84B.
    CrossrefPubMedGoogle Scholar
  • 2 Sherman CT, Litvack F, Grundfest W, Lee M, Hickey A, Chaux A, Kass R, Blanche C, Matloff J, Morgenstern L, Ganz W, Swan HJC, Forrester J. Coronary angioscopy in patients with unstable angina pectoris. N Engl J Med 1986; 315: 913-9.
    CrossrefPubMedGoogle Scholar
  • 3 Ambrose JA, Winters SL, Arora RR, Eng A, Riccio A, Gorlin R, Fuster V. Angiographic evaluation of coronary artery morphology in unstable angina. J Am Coll Cardiol 1986; 1: 472-8.
    PubMedGoogle Scholar
  • 4 Davies MJ, Thomas AC. Plaque fissuring – the cause of acute myocardial infarction, sudden ischemic death, and crescendo angina. Br Heart J 1985; 53: 363-73.
    CrossrefPubMedGoogle Scholar
  • 5 Telford AM, Wilson C. Trial of heparin versus atenolol in prevention of myocardial infarction in intermediate coronary syndrome. Lancet 1981; 1: 1225-8.
    PubMedGoogle Scholar
  • 6 Théroux P, Ouimet H, McCans J, Latour JG, Joly P, Lévy G, Pelletier E, Juneau M, Stasiak J, deGuise P, Pelletier GB, Rinzler D, Waters DD. Aspirin, heparin, or both to treat acute unstable angina. N Engl J Med 1988; 319: 1105-11.
    CrossrefPubMedGoogle Scholar
  • 7 The Risc Group.. Risk of myocardial infarction and death during treatment with low dose aspirin and intravenous heparin in men with unstable coronary artery disease. Lancet 1990; 336: 827-30.
    CrossrefPubMedGoogle Scholar
  • 8 Hirsh J. Heparin. N Engl J Med 1991; 324: 1565-74.
    CrossrefPubMedGoogle Scholar
  • 9 Markwardt F. Pharmacology of hirudin: one hundred years after the first report of the anticoagulant agent in medicinal leeches. Biomed Biochem Acta 1985; 44: 1007-13.
    PubMedGoogle Scholar
  • 10 Kaiser B, Markwardt F. Antithrombotic and haemorrhagic effects of the naturally occurring thrombin inhibitor hirudin. Folia Haematol Leipzig 1988; 115: 41-6.
    PubMedGoogle Scholar
  • 11 Bucha E, Nowak G, Markwardt F. Prevention of experimental coronary thrombosis by hirudin. Folia Haematol Leipzig 1988; 115: 52-8.
    PubMedGoogle Scholar
  • 12 Krupinski K, Breddin HK, Markwardt F, Haarmann W. Antithrombotic effects of three thrombin inhibitors in a rat model of laser-induced thrombosis. Haemostasis 1989; 19: 74-82.
    PubMedGoogle Scholar
  • 13 Komatsu Y, Misawa S, Sukesada A, Ohba Y, Hayashi H. CX-397, a novel recombinant hirudin analog having a hybrid sequence of hirudin variants-1 and -3. Biochem Biophys Res Commun 1993; 196: 773-9.
    CrossrefPubMedGoogle Scholar
  • 14 Hayashi H, Komatsu Y, Inoue Y, Sasaki M, Murata T, Fukazawa T, Goto Y, Misawa S, Takaku H, Furuya H, Matsuda H, Abe S. CX-397, a novel recombinant hirudin analog: (II) Physicochemical and antithrombotic properties. Thromb Haemost 1993; 69: 1302 (Abstract 2720).
    PubMedGoogle Scholar
  • 15 Romson JL, Haack DW, Lucchesi BR. Electrical induction of coronary artery thrombosis in the ambulatory canine: A model for in vivo evaluation of anti-thrombotic agents. Thromb Res 1980; 17: 841-53.
    CrossrefPubMedGoogle Scholar
  • 16 Romson JL, Haack DW, Abrams GD, Lucchesi BR. Prevention of occlusive coronary artery thrombosis by prostacyclin infusion in the dog. Circulation 1981; 64: 906-14.
    CrossrefPubMedGoogle Scholar
  • 17 Jergens AE, Turrentine MA, Kraus KH, Johnson GS. Buccal mucosa bleeding times of healthy dogs and of dogs in various pathologic states, including thrombocytopenia, uremia and von Willebrand’s disease. Am J Vet Res 1987; 48: 1337-42.
    PubMedGoogle Scholar
  • 18 Taguchi G. Unequal numbers of repetitions. In: System of experimental design. Two-way layout method. Clausing D ed. Michigan: American Supplier Institute; 1987: 36-40.
    Google Scholar
  • 19 Takiguchi Y, Asai F, Wada K, Hayashi H, Nakashima M. Antithrombotic effect of a novel recombinant hirudin analogue, CX-397, in a rat arterial thrombosis model. Br J Pharmacol 1995; 116: 3056-60.
    CrossrefPubMedGoogle Scholar
  • 20 Heras M, Chesebro JH, Webster MWI, Mruk JS, Grill DE, Penny WJ, Bowie EJW, Badimon L, Fuster V. Hirudin, heparin, and placebo during deep arterial injury in the pig. Circulation 1990; 82: 1476-84.
    CrossrefPubMedGoogle Scholar
  • 21 Agnelli G, Pascucci C, Cosmi B, Nenci GG. The comparative effects of recombinant hirudin (CGP 39393) and standard heparin on thrombus growth in rabbits. Thromb Haemost 1990; 63: 204-7.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 22 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.
    PubMedGoogle Scholar
  • 23 Weitz JI, Hudoba M, Massel D, Maraganore J, Hirsh J. Clot-bound thrombin is protected from inhibition by heparin-antithrombin III but is suceptible to inactivation by antithrombin III-independent inhibitors. J Clin Invest 1990; 86: 385-91.
    CrossrefPubMedGoogle Scholar
  • 24 Jackson CV, Crowe VG, Frank JD, Wilson HC, Coffman WJ, Utterback BG, Jakubowski JA, Smith GF. Pharmacological assessment of the anti-thrombotic activity of the peptide thrombin inhibitor, D-methylphenylalanyl-prolyl-arginal (GYKI-14766), in a canine model of coronary artery thrombosis. J Pharmacol Exp Ther 1992; 261: 546-52.
    PubMedGoogle Scholar
  • 25 Lynch JJ, Sitko GR, Lehman ED, Vlasuk GP. Primary prevention of coronary arterial thrombosis with the factor Xa inhibitor rTAP in a canine electrolytic injury model. Thromb Haemost 1995; 74: 640-5.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 26 Lindahl U, Thunberg L, Bäckström G, Riesenfeld J, Nordling K, Björk I. Extension and structural variability of the antithrombin-binding sequence in heparin. J Biol Chem 1984; 259: 12368-76.
    PubMedGoogle Scholar
  • 27 Carteaux JP, Gast A, Tschopp TB, Roux S. Activated clotting time as an appropriate test to compare heparin and direct thrombin inhibitors such as hirudin or Ro 46-6240 in experimental arterial thrombosis. Circulation 1995; 91: 1568-74.
    CrossrefPubMedGoogle Scholar
  • 28 Inoue Y, Komatsu Y, Satoh T, Uda K, Awaji T, Hayashi H. CX-397, a novel recombinant hirudin analog: (III) Study on the metabolism in rats. Thromb Haemost 1993; 69: 1303 (Abstract 2726).
    PubMedGoogle Scholar
  • 29 Märki MW, Wallis RB. The anticoagulant and antithrombotic properties of hirudins. Thromb Haemost 1990; 64: 344-8.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 30 Walenga JM, Hoppensteadt D, Koza M, Wallock M, Pifarre R, Fareed J. Laboratory assays for the evaluation of recombinant hirudin. Haemostasis 1991; 21 (Suppl. 01) 49-63.
    PubMedGoogle Scholar
  • 31 Zoldhelyi P, Webster MWI, Fuster V, Grill DE, Gaspar D, Edwards SJ, Cabot CF, Chesebro JH. Recombinant hirudin in patients with chronic, stable coronary artery disease. Safety, half-life, and effect on coagulation parameters. Circulation 1993; 88: 2015-22.
    CrossrefPubMedGoogle Scholar
  • 32 Tripodi A, Chantarangkul V, Arbini AA, Moia M, Mannucci PM. Effects of hirudin on activated partial thromboplastin time determined with ten different reagents. Thromb Haemost 1993; 70: 286-8.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 33 Monreal M, Monreal L, Gopegui RR, Espada Y, Angles AM, Monasterio J. Effects of two different doses of hirudin on APTT, determined with eight different reagents. Thromb Haemost 1995; 73: 219-22.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 34 Walenga JM, Hoppensteadt D, Koza M, Pifarre R, Fareed J. Comparative studies on various assays for the laboratory evaluation of r-hirudin. Semin Thromb Hemost 1991; 17: 103-12.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 35 Reid TJ, Alving BM. A quantitative thrombin time for determining levels of hirudin and Hirulog™. Thromb Haemost 1993; 70: 608-16.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 36 Nowak G, Bucha E. Quantitative determination of hirudin in blood and body fluids. Semin Thromb Hemost 1996; 22: 197-202.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 37 Gygax D, Botta L, Ehrat M, Graf P, Lefèvre G, Oroszlan P, Pfister C. Immunoassays in monitoring biotechnological drugs. Ther Drug Monit 1996; 18: 405-9.
    CrossrefPubMedGoogle Scholar
  • 38 Iwade K, Aosaki M, Ohki K, Saitoh K, Handa J, Nagashima H, Usui H, Hokari T, Uetsuka Y, Hosoda S. Changes in coagulation and fibrinolytic activity in unstable angina. Suppl to JAMA Southeast Asia 1993; 9: 10-4.
    PubMedGoogle Scholar
  • 39 Eisenberg PR, Sherman LA, Schectman K, Perez J, Sobel BE, Jaffe AS. Fibrinopeptide A: a marker of acute coronary thrombosis. Circulation 1985; 71: 912-8.
    CrossrefPubMedGoogle Scholar
  • 40 Théroux P, Latour JG, Léger Gauthier C, De Lara J. Fibrinopeptide A and platelet factor levels in unstable angina pectoris. Circulation 1987; 75: 156-62.
    CrossrefPubMedGoogle Scholar
  • 41 Merlini PA, Bauer KA, Oltrona L, Ardissino D, Cattaneo M, Belli C, Mannucci PM, Rosenberg RD. Persistent activation of coagulation mechanism in unstable angina and myocardial infarction. Circulation 1994; 90: 61-8.
    CrossrefPubMedGoogle Scholar