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DOI: 10.1055/s-0038-1649793
Characterisation of a Novel Series of Aprotinin-derived Anticoagulants
II. Comparative Antithrombotic Effects on Primary Thrombus Formation In VivoPublikationsverlauf
Received 03. Februar 1995
Accepted after resubmission 24. März 1995
Publikationsdatum:
06. Juli 2018 (online)
![](https://www.thieme-connect.de/media/10.1055-s-00035024/199502/lookinside/thumbnails/10-1055-s-0038-1649793-1.jpg)
Summary
Upon vascular damage platelet activation and blood coagulation are initiated. Interference at the initial level of the activation of the coagulation cascade can result in effective inhibition of thrombus formation. The in vivo antithrombotic properties of a series of bovine pancreatic trypsin inhibitor mutants (BPTI, aprotinin) 4C2, 7L22, 5L15, 5L15-PEG, 6L15 and 5L84, as described in the accompanying paper, with a combined inhibitory activity on factor Xa, factor VIIa-tissue factor complex, factor XIa and plasma kallikrein were compared to rTAP, r-hirudin, heparin and enoxaparin in a platelet rich thrombosis model in hamsters.
Platelet dependent thrombus deposition was quantified by dedicated image analysis after transillumination of the femoral vein to which a standardised vascular trauma was applied. After increasing intravenous bolus injections all tested agents, except for aprotinin, induced a dose dependent decrease of thrombus formation and a concomitant prolongation of the aPTT. From the linear correlation between these two parameters it was found that 5 out of the 6 tested aprotinin analogues, rTAP and r-hirudin completely inhibited thrombus formation at a therapeutical (2- to 3-fold) aPTT prolongation while 4C2, heparin and enoxaparin only inhibited thrombus formation for 40 to 50 percent at a 2-fold aPTT prolongation. Based on the calculated IC50 values for thrombus formation rTAP was found to be the most active compound in this model.
It is concluded that acceptable interference at the initial level of the blood coagulation, e. g. within a therapeutical aPTT prolongation, can significantly inhibit platelet deposition at a site of vascular injury.
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References
- 1 Stassen JM, Lambeir AM, Mathyssens G, Ripka WC, Nyström A, Sixma JJ, Vermylen J. Characterisation of a novel series of aprotinin-derived anticoagulants. I. In vitro and pharmacological properties Thromb Haemost 1995; 74: 646-654
- 2 Friedman M, Van den Bovenkamp GJ. The pathogenesis of a coronary thrombus. Am J Pathol 1966; 48: 19-44
- 3 Davies MJ, Thomas AC. Plaque fissuring: The cause of acute myocardial infarction, sudden ischaemic death, and crescendo angina. Br Heart J 1985; 53: 363-373
- 4 Fuster V, Chesebro JH. Mechanism of unstable angina. N Engl J Med 1986; 315: 1023-1025
- 5 Folts JD, Crowell EB, Rowe GG. Platelet aggregation in partially obstructed vessels and its elimination with aspirin. Circulation 1976; 54: 365-370
- 6 Willerson JT, Hillis DL, Winniford M, Buja LM. Speculation regarding mechanisms responsible lor acute ischemic disease syndromes. J Am Coll Cardiol 1986; 8: 245-250
- 7 Gold HK, Coller BS, Vasuda T, ballon JT, Guerrero JL, Leinbaeh LC, Ziskind AA, Collen D. Rapid and sustained coronary artery rceanalization with combined bolus injection of recombinant tissue-type plasminogen activator and a monoclonal antiplatelet GPIIb/llla antibody in a canine preparation. Circulation 1988; 77: 670-677
- 8 Chiang TM, Kang AH. Isolation and purification of collagen alpha 1(1) receptor from human platelet membrane. J Biol chem 1982; 257: 7581-7586
- 9 Sakariassen KS, Bolhuis PA, Sixma JJ. Human blood platelet adhesion to artery subendothelium is mediated by factor VIII-von Willcbrand factor bound to subendothelium. Nature 1978; 279: 636-638
- 10 Zwaal RF A. Membrane and lipid involvement in blood coagulation. Biochim Biophys Acta 1978; 515: 163-205
- 11 Osterud B, Rapaport SL. Activation of factor IX by the reaction product of tissue factor and factor VII: additional pathway for initiating blood coagulation. Proc Natl Acad Sci USA 1977; 74: 5260-5264
- 12 Zur M, Nemerson Y. The esterase activity of coagulation factor VII. Evidence for intrinsic activity of the zymogen J Biol Chem 1978; 253: 2203-2209
- 13 Bach RR. Initiation of coagulation by tissue factor. CRC Crit Rev Biochcm 1988; 23: 339-368
- 14 Drake TA, Morrissey JH, Edgington TS. Selective cellular expression of tissue factor in human tissues Implications for disorders of hemostasis and thrombosis. Am J Pathol 1989; 134: 1087-1097
- 15 Zwaal RF A, Bevers EM, Comfurius P. Platelets and coagulation. In: Blood coagulation. Zwaal RF A, Hcmker HC. (eds) New York: Elsevier Science Publishers; 1986. pp 141-169
- 16 Mann KG, Nesheim ME, Fraey PB. Non enzymatic cofactors: Factor V. In: Blood Coagulation. Zwaal RF A, Hemker HC. (eds) New York: Elsevier Science Publishers; 1993. pp 15-32
- 17 Scully MF. Kinetic considerations on the physiologic control of thrombin and factor Xa. Semin Thromb Haemost 1992; 18: 218-223
- 18 Lankiewicz MW, Bell WR. A unique circulating inhibitor with specificity for coagulation factor X. Am J Med 1992; 93: 343-346
- 19 Imura Y, Stassen JM, Collen D. Comparative antithrombotic effects of heparin, recombinant hirudin and argatroban in a hamster femoral vein platelet-rich mural thrombosis model. J Pharmacol Exp Thcr 1992; 261: 895-898
- 20 Zoldhelyi P, Fuster V, Chesebro JH. Antithrombins as conjunctive therapy in arterial thrombolysis. Cor Art Dis 1992; 3: 1003-1009
- 21 Theroux P, Latour JP. Anticoagulants and their use in acute ischemic syndrome. In: Textbook of interventional cardiology. Topol EJ. (ed) Philadelphia: W. B. Saunders; 1990. pp 24-48
- 22 Rapold HJ, Lu HR, Wu ZM, Nijs H, Collen D. Requirement of heparin for arterial and venous thrombolysis with recombinant tissue-type plasminogen activator. Blood 1991; 77: 1020-1024
- 23 Hirsh J. Heparin and low-molecular weight heparins. Cor Art Dis 1992; 3: 990-1002
- 24 Stassen JM, Rapold HJ, Vanlinthout I, Collen D. Comparative effect of enoxaparin and heparin on arterial and venous clot lysis with alteplase in dogs. Thromb Haemost 1993; 69: 454-459
- 25 Schaffer LW, Davidson JT, Vlasuk GP, Siegl PK. Antithrombotic efficacy of recombinant tick anticoagulant peptide. A potent inhibitor of coagulation factor Xa in a primate model of arterial thrombosis Circulation 1991; 84: 1741-1748
- 26 Markwardt F. Development of hirudin as an antithrombotic agent. Semin Thromb Haemost 1989; 15: 269-282
- 27 Neeper MP, Waxman L, Smith DE, Schulman CA, Sardana M, Ellis RW, Schaffer LW, Siegl PK, Vlasuk GP. Characterization of recombinant tick anticoagulant peptide. A highly selective inhibitor of blood coagulation factor Xa J Biol Chem 1990; 265: 17746-17752
- 28 Broze GJ. Tissue factor pathway inhibitor and the revised hypothesis of blood coagulation. Trends Cardiovasc Med 1992; 2: 72-77
- 29 Eisenberg PR. Development of novel anticoagulants for use as conjunctive therapy for thrombolysis. Cor Art Dis 1992; 3: 1010-1015
- 30 Eidt JF, Allison P, Noble S, Ashton J, Golino P, McNatt J, Buja LM, Willerson JT. Thrombin is an important mediator of platelet aggregation in stenosal canine coronary arteries with endothelial injury. J Clin Invest 1989; 84: 18-27
- 31 Heras M, Chesebro JH, Penny WJ, Bailey KR, Badimon E, 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: 647-665
- 32 Sitko GR, Ramjit DR, Stabilito H, Lehman D, Lynch JJ, Vlasuk GP. Conjunctive enhancement of enzymatic thrombolysis and prevention of thrombotic reocclusion with the selective factor Xa inhibitor, tick anticoagulant peptide. Comparison to hirudin and heparin in a canine model of acute coronary artery thrombosis Circulation 1992; 85: 805-815
- 33 De Macycr M, Lambeir AM, Laroche Y, Lasters I, Lauwereys M, Mathyssens G, Stanssens P, Ripka W, Vlasuk G. Recombinant Kiinitz inhibitors of coagulation proteases. I. Site specific mutagenesis. Thromb Haemost 1993; 69: 888 (abstract 1245)
- 34 De Maeyer M, Lambeir AM, Laroche Y, Lasters I, Lauwereys M, Mathyssens G, Stanssens P, Ripka W, Vlasuk G. Recombinant Kiinitz inhibitors of coagulation proteases. II. Random mutagenesis Thromb Haemost 1993; 69: 889 (abstract 1250)
- 35 Stanssens P, De Maeyer M, Lambeir AM, Laroche Y, Lauwereys M, Lasters I, Ripka WC. Design of inhibitors of serine proteases: Use of site selective and random mutagenesis to define the pharmacophore and conformational selectivity of Kunitz inhibitors towards factors Xa and VIIa. Submitted
- 36 Stockmans F, Deckmyn H, Gruwez J, Vcrmylen J, Acland R. Continuous quantitative monitoring of mural, platelet-dependent, thrombus kinetics in the crushed rat femoral vein. Thromb Haemost 1991; 65: 425-431
- 37 Imura Y, Stassen JM, Bunting S, Stockmans F, Collen D. Antithrombotic properties of L-cysteine, N-(mercaptoacetyl)-D-Tyr-Arg-Gly-Asp-sulfoxide (G4120) in a hamster platelet-rich femoral vein thrombosis model. Blood 1992; 80: 1247-1253
- 38 Laroche Y, Strome V, De Meutter J, Messens J, Lauwereys M. High-level secretion and very efficient isotopic labeling of thick anticoagulant peptide (TAP) expressed in the methlyotrophic yeast, Pischia pastoris. Bio/Technology 1994; 2: 1119-1124
- 39 Giles AR. Guidelines for the use of animals in biomedical research. Thromb Haemost 1987; 58: 1078-1084
- 40 Agnelli G, Pascucci C, Cosmi B, Nenci GG. Effects of hirudin and heparin on the binding of new fibrin to the thrombus in t-PA treated rabbits. Thromb Haemost 1991; 66: 592-597
- 41 Agnelli G, Renga C, Weitz JI, Nenci GG, Hirsh J. Sustained antithrombotic activity of hirudin after its plasma clearance: comparison with heparin. Blood 1992; 80: 960-965
- 42 Zoldhelyi P, Webster MW, 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: 2015-2022
- 43 Cannon CP, Maraganore JM, Loscalzo J, McAllister A, Eddings K, George D, Selwyn AP, Adelman B, Fox I, Braunwald E, Ganz P. Anticoagulant effects of hirulog, a novel thrombin inhibitor, in patients with coronary artery disease. Am J Cardiol 1993; 71: 778-782
- 44 Topol EJ, Bonan R, Jewitt D, Sigwart U, Kakkar VV, Rothman M, de Bono D, Ferguson J, Willerson JT, Strony J, Ganz P, Chen MD, Raymond R, Fox I, Maraganore JM, Adelman B. Use of a direct antithrombin, hirulog, in place of heparin during coronary angioplasty. Circulation 1993; 87: 1622-1629
- 45 Vlasuk GP, Ramjit D, Fujita T, Dunwiddie CT, Nutt EM, Smith DE, She-buski RJ. Comparison of the in vivo anticoagulant properties of standard heparin and the highly selective factor Xa inhibitors antistasin and tick anticoagulant peptide (TAP) in a rabbit model of venous thrombosis. Thromb Haemost 1991; 65: 257-262