Antibodies for inhibiting blood coagulation and methods of use thereof

Abstract

The invention includes antibodies that provide superior anti-coagulant activity by binding native human TF with high affinity and specificity. Antibodies of the invention can effectively inhibit blood coagulation in vivo. Antibodies of the invention can bind native human TF, either alone or present in a TF:VIIa complex, effectively preventing factor X binding to TF or that complex, and thereby reducing blood coagulation. Preferred antibodies of the invention specifically bind a conformational epitope predominant to native human TF, which epitope provides an unexpectedly strong antibody binding site.

1 . An antibody that binds native human tissue factor and does not substantially bind non-native tissue factor.  
     
     
         2 . An antibody of  claim 1  wherein the antibody has the binding specificity for native human tissue factor about equal to or greater than H36.D2.B7 [ATCC HB-12255].  
     
     
         3 . An antibody having the binding affinity for native human tissue factor about equal to or greater than H36.D2.B7 [ATCC HB-12255].  
     
     
         4 . An antibody having identifying characteristics of H36.D2.B7 [ATCC HB-12255].  
     
     
         5 . An antibody of  claim 1  wherein the antibody is H36.D2.B7 [ATCC HB-12255].  
     
     
         6 . An antibody that binds native tissue factor to form a complex whereby factor X binding to the complex is inhibited.  
     
     
         7 . An antibody of  claim 1  wherein the antibody is a monoclonal antibody.  
     
     
         8 . An antibody of  claim 1  that is a chimeric antibody.  
     
     
         9 . An antibody of  claim 8  that comprises a constant region of human origin.  
     
     
         10 . An antibody of  claim 1  that is a single chain antibody.  
     
     
         11 . An antibody that comprises a sequence that has at least about 70 percent sequence identity to SEQ ID NO:1.  
     
     
         12 . An antibody of  claim 11  that comprises a sequence represented by SEQ ID NO:2 or SEQ ID NO:4.  
     
     
         13 . An antibody that comprises hypervariable regions that have at least 90 percent sequence identity to SEQ ID NOS. 5 through 10 inclusive.  
     
     
         14 . An antibody of  claim 13  wherein the antibody comprises hypervariable regions represented by SEQ ID NOS. 5 through 10 inclusive.  
     
     
         15 . An isolated nucleic acid comprising a sequence encoding at least a portion of an antibody that binds native human tissue factor.  
     
     
         16 . The nucleic acid of  claim 15  wherein the monoclonal antibody is H36.D2.B7 [ATCC HB-12255].  
     
     
         17 . The nucleic acid of  claim 15  wherein the nucleic acid comprises SEQ ID NO:1 or SEQ ID NO:3.  
     
     
         18 . The nucleic acid of  claim 15  wherein the nucleic acid comprises a sequence that has at least about 70 percent sequence identity to SEQ ID NO:1 or SEQ ID NO:3.  
     
     
         19 . The nucleic acid of  claim 15  wherein the nucleic acid comprises sequences coding for antibody hypervariable regions that have at least 90 percent sequence identity to SEQ ID NOS. 5 through 10 inclusive.  
     
     
         20 . A nucleic acid comprising at least about 100 base pairs and that hybridizes to SEQ ID NO:1 or SEQ ID NO:3 under normal stringency conditions.  
     
     
         21 . A nucleic acid of  claim 20  wherein the nucleic acid hybridizes to SEQ ID NO:1 or SEQ ID NO:3 under high stringency conditions.  
     
     
         22 . A nucleic acid of  claim 15  wherein the nucleic acid comprises sequences that have at least 90 percent sequence identitity to SEQ ID NOS. 11 through 16 inclusive and code for hypervariable regions.  
     
     
         23 . A recombinant vector comprising the nucleic acid of  claim 15 , wherein the vector can express at least a portion of an antibody that binds native human tissue factor.  
     
     
         24 . A host cell comprising the vector of  claim 23 .  
     
     
         25 . A method of inhibiting blood coagulation in a mammal, comprising administering to the mammal an effective amount of an antibody capable of specifically binding native tissue factor and whereby the antibody complexes with native tissue factor, and factor X binding to the complex is inhibited.  
     
     
         26 . The method of  claim 25  wherein the complex further comprises factor VII/VIIa.  
     
     
         27 . The method of  claim 25  wherein the mammal is a human.  
     
     
         28 . The method of  claim 25  wherein the human is suffering from or suspected of having a thrombosis.  
     
     
         29 . The method of  claim 25  wherein the human is suffering from or susceptible to restenosis associated with an invasive medical procedure.  
     
     
         30 . The method of  claim 29  wherein the invasive medical procedure is angioplasty, endarterectomy, deployment of a stent, use of catheter, graft implantation or use of an arteriovenous shunt.  
     
     
         31 . The method of  claim 25  wherein the human is suffering from a thromboembolic condition associated with cardiovascular disease, an infectious disease, a neoplastic disease or use of a thrombolytic agent.  
     
     
         32 . The method of  claim 25  further comprising administering an anti-platelet composition, a thrombolytic composition or an anti-coagulant composition.  
     
     
         33 . The method of  claim 25  wherein the antibody is H36.D2.B7 [ATCC HB-12255].  
     
     
         34 . A method of reducing tissue factor levels in a mammal comprising: 
 administering to the mammal a therapeutically effective amount of an antibody capable of binding native tissue factor, the antibody linked covalently to a cell toxin or an effector molecule to provide complement-fixing ability and antibody-dependent cell-mediated cytotoxicity,    the antibody contacting cells expressing tissue factor to reduce tissue factor levels in the mammal.    
     
     
         35 . The method of  claim 34  wherein the cells expressing tissue factor are cancer cells, immune cells, or endothelial cells.  
     
     
         36 . A method of detecting tissue factor in a biological sample comprising: 
 contacting a biological sample with a monoclonal antibody of  claim 1  and analyzing the biological sample and monoclonal antibody for the presence of tissue factor in the biological sample.