Cardiovascular disease is the major cause of mortality and morbidity in the industrialized world. Thrombosis resulting from the activation of the blood coagulation cascade is the common etiology which underlies the three main categories that comprise this disease which includes: coronary artery disease leading to acute myocardial infarction (AMI), cerebrovascular disease leading to stroke and venous thrombosis which often leads to pulmonary embolism. There are only a limited number of therapies available to treat these diseases none of which are overwhelmingly effective. We propose in this phase II proposal to improve upon and evaluate the selective inhibitors of blood coagulation factors VIIa and Xa successfully engineered from mutants of a prototypical Kunitz serine protease inhibitor in the phase I portion of the grant. These studies will include site directed and random mutagenesis to improve the potency and selectivity of the inhibitors based on the three dimensional structural determination of an enzyme inhibitor complex and the full in vitro and in vivo evaluation of the inhibitory properties and antithrombotic effectiveness of selected inhibitors in established experimental whole animal models of thrombosis. It is anticipated that a novel and more clinically effective antithrombotic treatment will emerge as a result of these studies.
