Reaction of thrombin and proteinases of the fibrinolytic system with a mechanism-based inhibitor, 3,4-dihydro-3-benzyl-6-chloromethylcoumarin.
Thrombin, plasmin and tissue plasminogen activator (one- and two-chain forms) were examined with respect to their reaction with the suicide substrate, 3,4-dihydro-3-benzyl-6-chloromethylcoumarin, at 4 degrees C. The enzymes were irreversibly inhibited and the apparent second-order rate constants ki/Ki were 31,000, 316, 187 and 250 M-1.s-1, respectively. The extent of fibrin clot lysis induced by urokinase and two-chain tissue plasminogen activator was considerably decreased after treatment of these enzymes with the dihydrocoumarin derivative (molar excess of inhibitor over enzyme ranging from 6 to 21 for urokinase and 50 to 1500 for tissue plasminogen activator). This inhibitor has been tested as anticoagulant in human plasma and was effective at prolonging the prothrombin time from 12 to 40 s.
Mor
A
A
Laboratoire de Bioactivation des Peptides, Institut Jacques Monod, Paris, France.
Maillard
J
J
Favreau
C
C
Reboud-Ravaux
M
M
D001777:Blood Coagulation / Q000187:drug effects; D003374:Coumarins / Q000494:pharmacology*; D005341:Fibrinolysin / Q000037:antagonists & inhibitors*; D005342:Fibrinolysis; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D011480:Protease Inhibitors; D011517:Prothrombin Time; D013917:Thrombin / Q000037:antagonists & inhibitors*; D010959:Tissue Plasminogen Activator / Q000037:antagonists & inhibitors*; D014568:Urokinase-Type Plasminogen Activator / Q000037:antagonists & inhibitors*
D003374:Coumarins; D011480:Protease Inhibitors; C058018:3,4-dihydro-3-benzyl-6-chloromethylcoumarin; D013917:Thrombin; D010959:Tissue Plasminogen Activator; D005341:Fibrinolysin; D014568:Urokinase-Type Plasminogen Activator