Aprotinin is a competitive inhibitor of the factor VIIa-tissue factor complex.
A highly purified preparation of human plasma factor VIIa was submitted to chromogenic assays with S-2288 factors IXa, Xa, activated protein C and thrombin being absent. Factor VIIa alone or in the presence of calcium, kept its activity even in the presence of high concentrations of aprotinin, inhibition appeared only in the presence of a factor VIIa-tissue factor complex. A two-stage amidolytic assay using activation of purified factor X and hydrolysis of S-2765 chromogenic substrate by the generated Xa was used to show a competitive inhibition with a Ki value of 30 microM. Aprotinin had no effect on factor Xa amidolytic activity per se. The factor VIIa-tissue factor complex could be adsorbed to immobilized aprotinin and removed by a chaotropic ion like KSCN 3 M. The assays with the DFP inactivated VIIa-tissue factor complex proved that the interaction involved the active site of factor VIIa. The inhibition of the VIIa-tissue factor complex was demonstrated in a clotting assay using aprotinin enriched normal or factor VIII deficient plasma.
Chabbat
J
J
Centre National de Transfusion Sanguine, Paris, France.
Porte
P
P
Tellier
M
M
Steinbuch
M
M
D007611:Aprotinin / Q000737:chemistry / Q000494:pharmacology*; D001777:Blood Coagulation / Q000187:drug effects; D002863:Chromogenic Compounds; D004789:Enzyme Activation / Q000187:drug effects; D015942:Factor VIIa / Q000037:antagonists & inhibitors* / Q000378:metabolism; D005170:Factor X / Q000378:metabolism; D006801:Humans; D011485:Protein Binding; D013329:Structure-Activity Relationship; D013925:Thromboplastin / Q000037:antagonists & inhibitors* / Q000378:metabolism
D002863:Chromogenic Compounds; D005170:Factor X; D013925:Thromboplastin; D007611:Aprotinin; D015942:Factor VIIa