title: Fibrinolytic compromise by simultaneous administration of site-directed inhibitors of thrombin.
abstract: Newly developed synthetic and recombinant thrombin inhibitors possess strong anticoagulant effects. Despite these effects, interactions of these agents with enzymes in the fibrinolytic network result in the modulation of such proteases as t-PA, u-PA and streptokinase. The inhibitory spectrum of several thrombin inhibitors [D-Phe-Pro-Arg-H(GYKI 14166), D-MePhe-Pro-Arg-H(GYKI 14766), Boc-D-Phe-Pro-Arg-H (GYKI 14451), Ac-D-Phe-Pro-boroArg-OH (DuP 714), recombinant hirudin (r-Hir) and unfractionated porcine mucosal heparin complexed with antithrombin III (Heparin/AT-III)] was studied towards various serine proteases such as tissue plasminogen activator (t-PA), plasmin, plasminogen/streptokinase complex, urokinase and kallikrein. Aprotinin was also studied in the same systems as the thrombin inhibitors. All four tripeptide derivatives were found to inhibit t-PA, plasmin and plasminogen/streptokinase complex at micromolar concentrations (IC50: 0.57 mM-3.3 microM). Boc-D-Phe-Pro-Arg-H and Ac-D-Phe-Pro-boroArg-OH also inhibited urokinase, while Ac-D-Phe-Pro-boroArg-OH inhibited kallikrein as well (IC50: 0.15 mM-16 microM). In contrast, r-Hir and Heparin/AT-III did not inhibit any of these enzymes at millimolar concentrations (IC50 > or = 1 mM). Aprotinin inhibited plasmin, plasminogen/streptokinase complex and kallikrein at micromolar concentrations (IC50: 3.1-0.85 microM). In a rabbit thrombolysis model, where pre-formed clots are lysed by streptokinase, simultaneous administration of D-MePhe-Pro-Arg-H or Ac-D-Phe-Pro-boroArg-OH, at concentrations approximately 1 mumol/kg, i.v. resulted in complete inhibition of the fibrinolytic process. Aprotinin at 0.1 mumol/kg, i.v. produced similar inhibition. These results demonstrate that thrombin inhibitors may exert significant antiprotease actions against various fibrinolytic enzymes.
authors.affiliation: Department of Pharmacology, Loyola University Chicago, Stritch School of Medicine, Maywood, Illinois.
authors.forename: D
authors.initials: D
authors.lastname: Callas
authors.forename: P
authors.initials: P
authors.lastname: Bacher
authors.forename: O
authors.initials: O
authors.lastname: Iqbal
authors.forename: D
authors.initials: D
authors.lastname: Hoppensteadt
authors.forename: J
authors.initials: J
authors.lastname: Fareed
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000990:Antithrombin III / Q000737:chemistry / Q000494:pharmacology; D000991:Antithrombins / Q000494:pharmacology; D005342:Fibrinolysis / Q000187:drug effects*; D006493:Heparin / Q000737:chemistry / Q000494:pharmacology; D006629:Hirudins / Q000494:pharmacology; D008969:Molecular Sequence Data; D011480:Protease Inhibitors / Q000494:pharmacology; D011817:Rabbits; D011994:Recombinant Proteins / Q000494:pharmacology; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology; D013917:Thrombin / Q000037:antagonists & inhibitors*; D013997:Time Factors
chemical_list: D000991:Antithrombins; D006629:Hirudins; D011480:Protease Inhibitors; D011994:Recombinant Proteins; D015842:Serine Proteinase Inhibitors; D000990:Antithrombin III; D006493:Heparin; D013917:Thrombin