Summary
The substrate recognition region of tissue factor contains two residues, Lys165 and
Lys166, which are important for macromolecular substrate activation by the tissue
factor:factor VIIa complex. Replacement of these two residues with alanine in a soluble
version of human tissue factor resulted in a mutant, hTFAA, which can bind factor
VIIa but forms an enzymatically inactive complex. We found that hTFAA inhibits the
activity of guinea pig factor VIIa, allowing us to evaluate hTFAAs effects on thrombosis
and hemostasis in a guinea pig model of recurrent arterial thrombosis. In addition
to heparin, the effects of hTFAA were compared to active site inhibited factor IXa
(F.IXai) and factor Xa (F.Xai). We found that hTFAA, F.IXai and F.Xai were potent
antithrombotics and may possess a decreased risk of hemorrhage when compared to unfractionated
heparin. When administered at a dose that inhibited thrombosis by about 90%, hTFAA
neither affected cuticle bleeding nor the activated partial thromboplastin time, and
had only a modest effect on the prothrombin time. At equi-efficacious doses, F.IXai,
F.Xai and heparin prolonged bleeding times by 20% (p >0.5), 50% (p <0.05) and 100%
(p <0.01), respectively. In summary, our study demonstrates that, unlike heparin,
specific inhibitors of factors VIIa, IXa and Xa can produce antithrombotic effects
without or with only minimally disturbing normal hemostasis. The results further suggest
that factor VIIa and factor IXa are especially promising targets for antithrombotic
drug development.
Keywords
Tissue factor - factor IX - factor X - factor VII - anticoagulants - thrombosis