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Title: Thrombostatin FM compounds: direct thrombin inhibitors - mechanism of action in vitro and in vivo

Abstract

Novel pentapeptides called Thrombostatin FM compounds consisting mostly of D-isomers and unusual amino acids were prepared based upon the stable angiotensin converting enzyme breakdown product of bradykinin - RPPGF. These peptides are direct thrombin inhibitors prolonging the thrombin clotting time, activated partial thromboplastin time, and prothrombin time at ≥0.78, 1.6, and 1.6 μm, respectively. They competitively inhibit α-thrombin-induced cleavage of a chromogenic substrate at 4.4--8.2 μm. They do not significantly inhibit plasma kallikrein, factor (F) XIIa, FXIa, FIXa, FVIIa-TF, FXa, plasmin or cathepsin G. One form, FM19 [rOicPaF(p-Me)], blocks α-thrombin-induced calcium flux in fibroblasts with an IC50 of 6.9 ± 1.2 μm. FM19 achieved 100% inhibition of threshold α- or γ-thrombin-induced platelet aggregation at 8.4 ± 4.7 μm and 16 ± 4 μm, respectively. The crystal structure of thrombin in complex with FM19 shows that the N-terminal D-Arg retrobinds into the S1 pocket, its second residue Oic interacts with His-57, Tyr-60a and Trp-60d, and its C-terminal p-methyl Phe engages thrombin's aryl binding site composed of Ile-174, Trp-215, and Leu-99. When administered intraperitoneal, intraduodenal, or orally to mice, FM19 prolongs thrombin clotting times and delays carotid artery thrombosis. FM19, a low affinity reversible direct thrombin inhibitor, might be useful as anmore » add-on agent to address an unmet need in platelet inhibition in acute coronary syndromes in diabetics and others who with all current antiplatelet therapy still have reactive platelets.« less

Authors:
; ; ; ; ; ; ; ; ; ; ; ; ;  [1]
  1. Case Western
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
USDOE
OSTI Identifier:
1006579
Resource Type:
Journal Article
Journal Name:
J. Thromb. Haemost.
Additional Journal Information:
Journal Volume: 6; Journal Issue: (5) ; 05, 2008
Country of Publication:
United States
Language:
ENGLISH
Subject:
60 APPLIED LIFE SCIENCES; AFFINITY; AGGLOMERATION; AMINO ACIDS; ANGIOTENSIN; BRADYKININ; BREAKDOWN; CALCIUM; CATHEPSINS; CLEAVAGE; CRYSTAL STRUCTURE; ENZYME INHIBITORS; ENZYMES; FIBRINOLYSIN; FIBROBLASTS; IN VITRO; IN VIVO; KALLIKREIN; MICE; PEPTIDES; PLASMA; PROTHROMBIN; RESIDUES; SUBSTRATES; THERAPY; THROMBIN; THROMBOPLASTIN; THROMBOSIS

Citation Formats

Nieman, M T, Burke, F, Warnock, M, Zhou, Y, Sweigart, J, Chen, A, Ricketts, D, Lucchesi, B R, Chen, Z, Cera, E Di, Hilfinger, J, Kim, J S, Mosberg, H I, Schmaier, A H, Michigan), TSRL), and WU-MED). Thrombostatin FM compounds: direct thrombin inhibitors - mechanism of action in vitro and in vivo. United States: N. p., 2008. Web. doi:10.1111/j.1538-7836.2008.02937.x.
Nieman, M T, Burke, F, Warnock, M, Zhou, Y, Sweigart, J, Chen, A, Ricketts, D, Lucchesi, B R, Chen, Z, Cera, E Di, Hilfinger, J, Kim, J S, Mosberg, H I, Schmaier, A H, Michigan), TSRL), & WU-MED). Thrombostatin FM compounds: direct thrombin inhibitors - mechanism of action in vitro and in vivo. United States. https://doi.org/10.1111/j.1538-7836.2008.02937.x
Nieman, M T, Burke, F, Warnock, M, Zhou, Y, Sweigart, J, Chen, A, Ricketts, D, Lucchesi, B R, Chen, Z, Cera, E Di, Hilfinger, J, Kim, J S, Mosberg, H I, Schmaier, A H, Michigan), TSRL), and WU-MED). 2008. "Thrombostatin FM compounds: direct thrombin inhibitors - mechanism of action in vitro and in vivo". United States. https://doi.org/10.1111/j.1538-7836.2008.02937.x.
@article{osti_1006579,
title = {Thrombostatin FM compounds: direct thrombin inhibitors - mechanism of action in vitro and in vivo},
author = {Nieman, M T and Burke, F and Warnock, M and Zhou, Y and Sweigart, J and Chen, A and Ricketts, D and Lucchesi, B R and Chen, Z and Cera, E Di and Hilfinger, J and Kim, J S and Mosberg, H I and Schmaier, A H and Michigan) and TSRL) and WU-MED)},
abstractNote = {Novel pentapeptides called Thrombostatin FM compounds consisting mostly of D-isomers and unusual amino acids were prepared based upon the stable angiotensin converting enzyme breakdown product of bradykinin - RPPGF. These peptides are direct thrombin inhibitors prolonging the thrombin clotting time, activated partial thromboplastin time, and prothrombin time at ≥0.78, 1.6, and 1.6 μm, respectively. They competitively inhibit α-thrombin-induced cleavage of a chromogenic substrate at 4.4--8.2 μm. They do not significantly inhibit plasma kallikrein, factor (F) XIIa, FXIa, FIXa, FVIIa-TF, FXa, plasmin or cathepsin G. One form, FM19 [rOicPaF(p-Me)], blocks α-thrombin-induced calcium flux in fibroblasts with an IC50 of 6.9 ± 1.2 μm. FM19 achieved 100% inhibition of threshold α- or γ-thrombin-induced platelet aggregation at 8.4 ± 4.7 μm and 16 ± 4 μm, respectively. The crystal structure of thrombin in complex with FM19 shows that the N-terminal D-Arg retrobinds into the S1 pocket, its second residue Oic interacts with His-57, Tyr-60a and Trp-60d, and its C-terminal p-methyl Phe engages thrombin's aryl binding site composed of Ile-174, Trp-215, and Leu-99. When administered intraperitoneal, intraduodenal, or orally to mice, FM19 prolongs thrombin clotting times and delays carotid artery thrombosis. FM19, a low affinity reversible direct thrombin inhibitor, might be useful as an add-on agent to address an unmet need in platelet inhibition in acute coronary syndromes in diabetics and others who with all current antiplatelet therapy still have reactive platelets.},
doi = {10.1111/j.1538-7836.2008.02937.x},
url = {https://www.osti.gov/biblio/1006579}, journal = {J. Thromb. Haemost.},
number = (5) ; 05, 2008,
volume = 6,
place = {United States},
year = {2008},
month = {4}
}