Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1993 Mar 1;290(Pt 2):309–312. doi: 10.1042/bj2900309

Benzyloxycarbonyl-D-Phe-Pro-methoxypropylboroglycine: a novel inhibitor of thrombin with high selectivity containing a neutral side chain at the P1 position.

G Claeson 1, M Philipp 1, E Agner 1, M F Scully 1, R Metternich 1, V V Kakkar 1, T DeSoyza 1, L H Niu 1
PMCID: PMC1132273  PMID: 8452516

Abstract

Thrombin, the blood-clotting enzyme, is a serine proteinase with trypsin-like specificity and is able to cleave Arg-Xaa peptide bonds but only in a very limited number of substrates (and sites therein). For the prevention and treatment of thrombosis the control of thrombin activity is a key target, and a variety of synthetic inhibitors have been introduced recently, all of which have a positive charge at the P1 site. We report the synthesis of the first example of a new class of inhibitor containing a neutral side chain at the P1 site, the peptide benzyloxycarbonyl-D-Phe-Pro- methoxypropylboroglycine. The peptide is a potent inhibitor of thrombin [Ki (limiting) = 7 nM] and is highly selective for its target enzyme in respect of other serine proteinases. This may be expected to confer considerable advantage in terms of specificity of action and reduced toxicity over conventional, positively charged, inhibitors.

Full text

PDF
309

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bajusz S., Barabás E., Tolnay P., Széll E., Bagdy D. Inhibition of thrombin and trypsin by tripeptide aldehydes. Int J Pept Protein Res. 1978 Oct;12(4):217–221. doi: 10.1111/j.1399-3011.1978.tb02889.x. [DOI] [PubMed] [Google Scholar]
  2. Berliner L. J., Shen Y. Y. Physical evidence for an apolar binding site near the catalytic center of human alpha-thrombin. Biochemistry. 1977 Oct 18;16(21):4622–4626. doi: 10.1021/bi00640a015. [DOI] [PubMed] [Google Scholar]
  3. Bode W., Mayr I., Baumann U., Huber R., Stone S. R., Hofsteenge J. The refined 1.9 A crystal structure of human alpha-thrombin: interaction with D-Phe-Pro-Arg chloromethylketone and significance of the Tyr-Pro-Pro-Trp insertion segment. EMBO J. 1989 Nov;8(11):3467–3475. doi: 10.1002/j.1460-2075.1989.tb08511.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bode W., Turk D., Karshikov A. The refined 1.9-A X-ray crystal structure of D-Phe-Pro-Arg chloromethylketone-inhibited human alpha-thrombin: structure analysis, overall structure, electrostatic properties, detailed active-site geometry, and structure-function relationships. Protein Sci. 1992 Apr;1(4):426–471. doi: 10.1002/pro.5560010402. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bone R., Shenvi A. B., Kettner C. A., Agard D. A. Serine protease mechanism: structure of an inhibitory complex of alpha-lytic protease and a tightly bound peptide boronic acid. Biochemistry. 1987 Dec 1;26(24):7609–7614. doi: 10.1021/bi00398a012. [DOI] [PubMed] [Google Scholar]
  6. Farr-Jones S., Smith S. O., Kettner C. A., Griffin R. G., Bachovchin W. W. Crystal versus solution structure of enzymes: NMR spectroscopy of a peptide boronic acid-serine protease complex in the crystalline state. Proc Natl Acad Sci U S A. 1989 Sep;86(18):6922–6924. doi: 10.1073/pnas.86.18.6922. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fenton J. W., 2nd, Ofosu F. A., Moon D. G., Maraganore J. M. Thrombin structure and function: why thrombin is the primary target for antithrombotics. Blood Coagul Fibrinolysis. 1991 Feb;2(1):69–75. [PubMed] [Google Scholar]
  8. Fenton J. W., 2nd Regulation of thrombin generation and functions. Semin Thromb Hemost. 1988 Jul;14(3):234–240. doi: 10.1055/s-2007-1002783. [DOI] [PubMed] [Google Scholar]
  9. Hauptmann J., Markwardt F. Pharmacologic aspects of the development of selective synthetic thrombin inhibitors as anticoagulants. Semin Thromb Hemost. 1992;18(2):200–217. doi: 10.1055/s-2007-1002426. [DOI] [PubMed] [Google Scholar]
  10. Kaiser B., Hauptmann J., Markwardt F. Untersuchungen zur Pharmakodynamik synthetischer Thrombininhibitoren vom Typ basisch substituierter N alpha-arylsulfonylierter Phenylalaninamide. Pharmazie. 1987 Feb;42(2):119–121. [PubMed] [Google Scholar]
  11. Kettner C., Mersinger L., Knabb R. The selective inhibition of thrombin by peptides of boroarginine. J Biol Chem. 1990 Oct 25;265(30):18289–18297. [PubMed] [Google Scholar]
  12. Kettner C., Shaw E. D-Phe-Pro-ArgCH2C1-A selective affinity label for thrombin. Thromb Res. 1979;14(6):969–973. doi: 10.1016/0049-3848(79)90014-8. [DOI] [PubMed] [Google Scholar]
  13. Mattson C., Eriksson E., Nilsson S. Anti-coagulant and anti-thrombotic effects of some protease inhibitors. Folia Haematol Int Mag Klin Morphol Blutforsch. 1982;109(1):43–51. [PubMed] [Google Scholar]
  14. Ni F., Konishi Y., Frazier R. B., Scheraga H. A., Lord S. T. High-resolution NMR studies of fibrinogen-like peptides in solution: interaction of thrombin with residues 1-23 of the A alpha chain of human fibrinogen. Biochemistry. 1989 Apr 4;28(7):3082–3094. doi: 10.1021/bi00433a052. [DOI] [PubMed] [Google Scholar]
  15. Philipp M., Maripuri S. Inhibition of subtilisin by substituted arylboronic acids. FEBS Lett. 1981 Oct 12;133(1):36–38. doi: 10.1016/0014-5793(81)80465-6. [DOI] [PubMed] [Google Scholar]
  16. Rae I. D., Scheraga H. A. H and 13C nuclear magnetic resonance spectra of some peptides with fibrinogen-like reactivity. Int J Pept Protein Res. 1979 Mar;13(3):304–314. doi: 10.1111/j.1399-3011.1979.tb01884.x. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES