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Thromb Haemost 1992; 67(03): 325-330
DOI: 10.1055/s-0038-1648441
Original Articles
Schattauer GmbH Stuttgart

In Vitro Inhibition of Blood Coagulation by Tripeptide Aldehydes - A Retrospective Screening Study Focused on the Stable D-MePhe-Pro-Arg-H · H2SO4

Daniel Bágdy
The Institute for Drug Research, Budapest, Hungary
,
Èva Barabás
The Institute for Drug Research, Budapest, Hungary
,
Sándor Bajusz
The Institute for Drug Research, Budapest, Hungary
,
Erzsébet Széll
The Institute for Drug Research, Budapest, Hungary
› Author Affiliations
Further Information

Publication History

Received 03 September 1990

Accepted after revision 29 September 1991

Publication Date:
03 July 2018 (online)

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Summary

A series of peptide aldehydes synthetized in our institute during the last 15 years were screened to detect their inhibitory effect on blood coagulation. Simple conventional clotting assays, platelet function tests and fibrinolytic methods were used to evaluate the inhibitory potency of the compounds in complex clotting systems as well as their supposed antifibrinolytic effect in vitro. Special attention was paid to the possible interactions with blood cells and plasma proteins, and to the functional stability of the inhibitors in several tissue homogenates. D-Phe-Pro-Arg-H (GYKI-14166, RGH-2958), Boc-D-Phe-Pro-Arg-H (GYKI-14451) and D-MePhe-Pro-Arg-H (GYKI-14766) were found to be the most potent inhibitors. The peptide aldehydes via formation of reversible complexes with thrombin impede the enzyme to react with the coagulation factors, platelet membrane and vessel wall. The compounds inhibit platelet aggregation induced by thrombin specifically without changing the sensitivity of platelets to other inducers. D-Phe-Pro-Arg-H and D-MePhe-Pro-Arg-H showed no antifibrinolytic effect. D-MePhe-Pro-Arg-H and Boc-D-Phe-Pro-Arg-H proved to be stable in dry state for years and in solution at room temperature for several days. The anticoagulant activity of the compounds was declared in NIH antithrombin units.

 

  • References

  • 1 Blomback B, Blomback M, Olsson P, Svendsen L, Aberg G. Synthetic peptide with anticoagulant and vasodilatory activity. Scand J Clin Lab Invest 1969; 24 Suppl. (107) 59-64
    PubMedSearch in Google Scholar
  • 2 Dorman LC, Cheng RC, Marshall FN. Mechanism of thrombin action on fibrinogen: activity of thrombin towards alpha(A)-fibrinogen peptides. In: Chemistry and Biology of Peptides. Meienhofer J. (ed). Ann Arbor Sci. Publ. Inc.; Ann Arbor, MI: 1972. pp 455-459
    Search in Google Scholar
  • 3 Geratz JD. Synthetic low molecular weight inhibitors of thrombin. Folia Haematol (Leipz.) 1972; 98: 455-470
    PubMedSearch in Google Scholar
  • 4 Markwardt F. Synthetic low molecular thrombin inhibitors. New concept of anticoagulants?. Haemostasis 1984; 3: 182-202
    PubMedSearch in Google Scholar
  • 5 Okamoto S, Hijikata A, Kinjo K, Kikumoto R, Ohkubo K, Tono-mura S, Tamao Y. A novel series of synthetic thrombin inhibitors having extremely potent and highly selective action. Kobe J Med Sci 1975; 21: 43-51
    PubMedSearch in Google Scholar
  • 6 Bajusz S, Barabas E, Szell E, Bagdy D. Peptide aldehyde inhibitors of the fibrinogen-thrombin reaction. In: Peptides - Chemistry, Structure and Biology. Meienhofer J. (ed). Ann Arbor Sci. Publ. Inc.; Ann Arbor, MI,: 1975. pp 603-608
    Search in Google Scholar
  • 7 Bajusz S, Barabas E, Tolnay P, Szell E, Bagdy D. Inhibition of thrombin and trypsin by peptide aldehydes. Int J Pept Protein Res 1978; 12: 217-221
    PubMedSearch in Google Scholar
  • 8 Gaffney PJ, Miller-Andersson M, Kirkwood TBL. Unreliability of chromogenic substrates for assay of clotting activity of thrombin. Haemostasis 1978; 7: 109-112
    PubMedSearch in Google Scholar
  • 9 Gaffney PJ, Philo RD. A commentary of new methodology in haemostasis using chromogenic substrates. In: Perspectives in Haemostasis. Fareed J, Messmore HL, Fenton II JW, Brinkhous KM. (eds). Pergamon Press; New York: 1981. pp 405-417
    Search in Google Scholar
  • 10 Bagdy D, Tolnay P. The action of some proteinase inhibitors from plants on blood clotting. Excerpta Med Int Congr Ser. 1962 No. 48, 21/612.115.3
    PubMedSearch in Google Scholar
  • 11 Aoyagi T, Takeuchi T, Matsuzaki A, Kawamura K, Kondo S, Hamada M, Maeda K, Umezawa H. Leupeptins, new protease inhibitors from actinomycetes. J Antibiot (Tokyo) 1969; 22: 283-286
    CrossrefPubMedSearch in Google Scholar
  • 12 Bajusz S, Szell E, Barabas E, Bagdy D. US Patent No. 4,399,065 (1983)
    PubMed
  • 13 Bajusz S, Szell E, Barabas E, Bagdy D. US Patent No. 4,478,745 (1984)
    PubMed
  • 14 Bajusz S, Szell E, Bagdy D, Dioszegi M, Fittler Zs, Jozsa F, Horvath Gy, Tomori E. US Patent No. 4,703,036 (1987)
    PubMed
  • 15 Bajusz S, Szell E, Bagdy D, Barabas E, Horvath Gy, Dioszegi M, Fittler Zs, Szabo G, Juhasz A, Szilagyi G. Highly active and selective anticoagulants: D-Phe-Pro-Arg-H, a free tripeptide prone to spontaneous inactivation, and its stable N-methyl derivative, D-MePhe-Pro-Arg-H. J Med Chem 1990; 33: 1729-1735
    CrossrefPubMedSearch in Google Scholar
  • 16 Bagdy D, Barabas E, Szell E, Bajusz S. Über die biochemische Pharmakologie einiger Tripeptidaldehyde. Folia Haematol (Leipz) 1982; 109: 22-32
    PubMedSearch in Google Scholar
  • 17 Bagdy D, Bajusz S, Rabloczky Gy. RGH-2958 (GYKI-14166) An updated monograph. Drugs of the Future 1985; 10 (10) 829-834
    CrossrefPubMedSearch in Google Scholar
  • 18 Mattson Ch, Eriksson E, Nilsson S. Anticoagulant and antithrombotic effects of some proteinase inhibitors. Folia Haematol (Leipz) 1982; 109: 43-51
    PubMedSearch in Google Scholar
  • 19 Witting JI, Pouliott Cr, Catalfamo JL, Fareed J, Fenton II JW. Thrombin inhibition with dipeptidyl argininals. Thromb Res 1988; 50: 461-468
    CrossrefPubMedSearch in Google Scholar
  • 20 Fareed J, Messmore HL, Kindel G, Balis JU. Inhibition of serine proteinases by low molecular weight peptides and their derivatives. Ann NY Acad Sci USA 1981; 370: 765-784
    CrossrefPubMedSearch in Google Scholar
  • 21 Tremoli E, Morazzoni G, Maderna P, Colli S, Paoletti R. Studies on the antithrombotic action of Boc-D-Phe-Pro-Arg-H (GYKI 14451). Thromb Res 1981; 23: 549-553
    CrossrefPubMedSearch in Google Scholar
  • 22 Bagdy D, Barabas E, Kazi E, Bereznay T. Hungarian Patents No. 155,833 (1965), 155,996 (1966)
    PubMed
  • 23 Aoki N, Naito K, Yoshida N. Inhibition of platelet aggregation by proteinase inhibitors. Possible involvement of proteinases in platelet aggregation. Blood 1978; 52: 1-12
    CrossrefPubMedSearch in Google Scholar
  • 24 Astrup T, Müllertz S. The fibrin plate method for estimating fibrinolytic activity. Arch Biochem Biophys 1952; 40: 346-351
    CrossrefPubMedSearch in Google Scholar
  • 25 Born GVR, Cross MJ. The aggregation of blood platelets. J Physiol (London) 1963; 168: 178-193
    CrossrefPubMedSearch in Google Scholar
  • 26 Acki N, von Kaulla KN. Potentiation by synthetic fibrinolytic agents of vascular activator and urokinase-induced lysis of human plasma clots as demonstrated by thrombelastography. Thromb Diathes Haemorrh (Stuttgart) 1972; 27: 246-251
    PubMedSearch in Google Scholar
  • 27 Verstraete M, Verwilghen R. Haematological disorders. In Drug Treatment. Principles and Practice of Clinical Pharmacology and Therapeutics. Avery GS. (ed) 2nd ed. Churchill-Livingstone; Edinburgh: 1980. p 919
    Search in Google Scholar
  • 28 The United States Pharmacopeia Twentieth Revision. XX. United States Pharmacopeial Convention Inc.; Rockville, MD: 1980. pp 365-366
  • 29 Koväcs A, Barabas E, Aränyi P, Rabloczky Gy. Thrombin inhibitor (GYKI-14766) prevents thrombin induced vascular effects. Eur J Pharmacol 1990; 183: 1844-1845
    PubMedSearch in Google Scholar
  • 30 Markwardt F. Pharmacological control of blood coagulation by synthetic, low molecular weight inhibitors of clotting enzymes. A new concept of anticoagulants. Trends Pharm Sei 1980; 1: 152-157
    PubMedSearch in Google Scholar