title: Novel peptides that inhibit heparanase activation of the coagulation system.
abstract: Heparanase is implicated in cell invasion, tumour metastasis and angiogenesis. It forms a complex and enhances the activity of the blood coagulation initiator - tissue factor (TF). We describe new peptides derived from the solvent accessible surface of TF pathway inhibitor 2 (TFPI-2) that inhibit the heparanase procoagulant activity. Peptides were evaluated in vitro by measuring activated coagulation factor X levels and co-immunoprecipitation. Heparanase protein and/or lipopolysaccharide (LPS) were injected intra-peritoneally and inhibitory peptides were injected subcutaneously in mouse models. Plasma was analysed by ELISA for thrombin-antithrombin complex (TAT), D-dimer as markers of coagulation activation, and interleukin 6 as marker of sepsis severity. Peptides 5, 6, 7, 21 and 22, at the length of 11-14 amino acids, inhibited heparanase procoagulant activity but did not affect TF activity. Injection of newly identified peptides 5, 6 and 7 significantly decreased or abolished TAT plasma levels when heparanase or LPS were pre-injected, and inhibited clot formation in an inferior vena cava thrombosis model. To conclude, the solvent accessible surface of TFPI-2 first Kunitz domain is involved in TF/heparanase complex inhibition. The newly identified peptides potentially attenuate activation of the coagulation system induced by heparanase or LPS without predisposing to significant bleeding tendency.
authors.forename: E
authors.initials: E
authors.lastname: Axelman
authors.forename: I
authors.initials: I
authors.lastname: Henig
authors.forename: Y
authors.initials: Y
authors.lastname: Crispel
authors.forename: J
authors.initials: J
authors.lastname: Attias
authors.forename: J-P
authors.initials: JP
authors.lastname: Li
authors.forename: B
authors.initials: B
authors.lastname: Brenner
authors.forename: I
authors.initials: I
authors.lastname: Vlodavsky
authors.affiliation: Yona Nadir, MD, PhD, Thrombosis and Hemostasis Unit, Rambam Health Care Campus, Haifa, Israel, Tel: +972 4 8453520, Fax: +972 4 8543886, Email: ynadir@netvision.net.il.
authors.forename: Y
authors.initials: Y
authors.lastname: Nadir
mesh_terms: D000818:Animals; D001777:Blood Coagulation / Q000187:drug effects; D004195:Disease Models, Animal; D004789:Enzyme Activation / Q000187:drug effects; D005170:Factor X / Q000378:metabolism; D005338:Fibrin Fibrinogen Degradation Products / Q000378:metabolism; D005966:Glucuronidase / Q000037:antagonists & inhibitors* / Q000235:genetics; D006023:Glycoproteins / Q000008:administration & dosage*; D057809:HEK293 Cells; D006801:Humans; D015850:Interleukin-6 / Q000378:metabolism; D008070:Lipopolysaccharides / Q000276:immunology; D051379:Mice; D008813:Mice, Inbred ICR; D018345:Mice, Knockout; D010446:Peptide Fragments / Q000008:administration & dosage*; D013917:Thrombin / Q000378:metabolism; D013925:Thromboplastin / Q000378:metabolism; D013927:Thrombosis / Q000097:blood* / Q000188:drug therapy; D014682:Vena Cava, Inferior / Q000378:metabolism*; D000099175:Heparanase
chemical_list: D005170:Factor X; D005338:Fibrin Fibrinogen Degradation Products; D005966:Glucuronidase; D006023:Glycoproteins; D015850:Interleukin-6; D008070:Lipopolysaccharides; D010446:Peptide Fragments; D013917:Thrombin; D013925:Thromboplastin; D000099175:Heparanase; C036309:fibrin fragment D; C021240:tissue-factor-pathway inhibitor 2
keywords: Heparanase; TFPI-2; anticoagulant; peptides; tissue factor