Antibodies Against Tissue Factor Pathway Inhibitor

Abstract

The invention relates to antibodies that specifically bind to tissue factor pathway inhibitor (TFPI) and that reduce the clotting time of blood. Such antibodies have utility in the treatment of subjects with a coagulopathy.

1 . A monoclonal antibody that is capable of specifically binding the K2 domain of TFPI, wherein said antibody is capable of specifically binding an epitope comprising residue R17 of SEQ ID NO: 2. 
     
     
         2 . The monoclonal antibody of  claim 1 , wherein the epitope of said antibody comprises one or more residues selected from the group consisting of E10, E11, D12, P13, R17, Y19, T21, Y23, F24, N26, Q28, Q31, C32, E33, R34, K36, and L50 of SEQ ID NO: 2. 
     
     
         3 . The monoclonal antibody of  claim 1 , wherein a light chain of said antibody comprises amino acid residues:
 E, in the position corresponding to position 31,   S, in the position corresponding to position 32,   D, in the position corresponding to position 33,   Y, in the position corresponding to position 37,   A, in the position corresponding to position 96,   T, in the position corresponding to position 97, and   F, in the position corresponding to position 99   of SEQ ID NO: 15;   and wherein a heavy chain of said antibody comprises amino acid residues:   N, in the position corresponding to position 31,   R, in the position corresponding to position 53,   S, in the position corresponding to position 54,   Y, in the position corresponding to position 57,   Y, in the position corresponding to position 59,   F, in the position corresponding to position 60,   P, in the position corresponding to position 61,   D, in the position corresponding to position 62,   Q, in the position corresponding to position 65,   Y, in the position corresponding to position 102,   D, in the position corresponding to position 103, and   D, in the position corresponding to position 106   of SEQ ID NO 18.   
     
     
         4 . The monoclonal antibody of  claim 3 , wherein said heavy chain further comprises an S in the position corresponding to position 52 of SEQ ID NO: 18. 
     
     
         5 . The monoclonal antibody of  claim 4 , wherein said light chain further comprises an H in the position corresponding to position 98 of SEQ ID NO: 15 and said heavy chain further comprises an S in the position corresponding to position 56 of SEQ ID NO: 18. 
     
     
         6 . The monoclonal antibody of  claim 1 , wherein a heavy chain of said antibody comprises:
 a CDR1 sequence of amino acids 31 to 35 (NYAMS) of SEQ ID NO: 18, wherein one of these amino acids may be substituted by a different amino acid; and/or a CDR2 sequence of amino acids 50 to 66 (TISRSGSYSYFPDSVQG) of SEQ ID NO: 18, wherein one, two or three of these amino acids may be substituted by a different amino acid; and/or   a CDR3 sequence of amino acids 99 to 110 (LGGYDEGDAMDS) of SEQ ID NO: 18, wherein one, two or three of these amino acids may be substituted by a different amino acid.   
     
     
         7 . The monoclonal antibody of  claim 1 , wherein the light chain of said antibody comprises:
 a CDR1 sequence of amino acids 24 to 39 (KSSQSLLESDGKTYLN) of SEQ ID NO: 15, wherein one, two or three of these amino acids may be substituted with a different amino acid; and/or   a CDR2 sequence of amino acids 55 to 61 (LVSILDS) of SEQ ID NO: 15, wherein one or two of these amino acids may be substituted with a different amino acid; and/or   a CDR3 sequence of amino acids 94 to 102 (LQATHFPQT) of SEQ ID NO: 15, wherein one or two of these amino acids may be substituted with a different amino acid.   
     
     
         8 . The monoclonal antibody according to  claim 1 , wherein said antibody has a binding affinity K D  value of less than 0.8 nM for TFPI, as determined by using surface plasmon resonance. 
     
     
         9 . The monoclonal antibody according to  claim 1 , wherein said antibody has a binding affinity K D  value of less than 1×10 31 10 M for TFPI, as determined by using surface plasmon resonance. 
     
     
         10 . The monoclonal antibody according to  claim 1 , wherein the monoclonal antibody is capable of binding the K2 domain of TFPI with a higher affinity than mAb2974. 
     
     
         11 . A monoclonal antibody according to  claim 1  that is capable of neutralising the TFPI inhibition of membrane-bound FVIIa/TF/FXa by at least 55% as measured in an FVIIa/TF/FXa inhibitor assay, when TFPI is saturated with said antibody. 
     
     
         12 . The monoclonal antibody according to  claim 1 , wherein the monoclonal antibody is a humanized antibody. 
     
     
         13 . The monoclonal antibody according to  claim 1 , wherein the monoclonal antibody is a human antibody. 
     
     
         14 . The monoclonal antibody according to  claim 1 , wherein the monoclonal antibody is a chimeric antibody. 
     
     
         15 . The monoclonal antibody according to  claim 1 , wherein the isotype of said antibody is IgG. 
     
     
         16 . The monoclonal antibody according to  claim 1 , wherein said isotype is selected from the group consisting of IgG1, IgG2, and IgG4. 
     
     
         17 . The monoclonal antibody of  claim 1 , wherein the monoclonal antibody is selected from the group consisting of a Fab fragment, a F(ab′) 2  fragment, a Fab′ fragment, a Fd fragment, a Fv fragment or a dAb fragment. 
     
     
         18 . A eukaryotic cell which expresses the monoclonal antibody, or a fragment thereof, according to  claim 1 . 
     
     
         19 . A pharmaceutical composition comprising an antibody according to  claim 1  and a pharmaceutically acceptable carrier or diluent. 
     
     
         20 . A method of treating a subject with a coagulopathy, comprising administering to said subject the monoclonal antibody according to  claim 1 .