Coagulation factor-targeting to tlt-1 on activated platelets

Abstract

The current invention relates to: procoagulant proteins which may, for example, be fusion proteins or chemical conjugates; methods of producing said procoagulant proteins; polynucleotides that encode said fusion proteins and cells that expresses them. Furthermore, the current invention relates to procoagulant proteins for use as a medicament. Individuals that have a coagulopathy, such as haemophilia A and B with or without inhibitors, may be treated with the procoagulant proteins of the current invention.

1 - 17 . (canceled) 
     
     
         18 . A procoagulant protein comprising a coagulation factor covalently attached to a monoclonal antibody or a fragment thereof, the monoclonal antibody or fragment thereof capable of specifically binding TLT-1 or a fragment or a variant thereof. 
     
     
         19 . The procoagulant protein according to  claim 18 , wherein the coagulation factor is a serine protease or a derivative thereof. 
     
     
         20 . The procoagulant protein according to  claim 19 , wherein the serine protease is a FVII polypeptide. 
     
     
         21 . The procoagulant protein according to  claim 19 , wherein the serine protease is a FIX polypeptide. 
     
     
         22 . The procoagulant protein according to  claim 18 , wherein the monoclonal antibody or fragment thereof is a Fab fragment. 
     
     
         23 . The procoagulant protein according to  claim 18 , the monoclonal antibody or fragment thereof having an epitope, wherein the epitope comprises one or more residues selected from the group consisting of V17, Q18, C19, H20, Y21, R22, L23, Q24, D25, V26, K27, A28, L63, G64, G65, G66, L67, L68, G89, A90, R91, G92, P93, Q94, I95, and L96 of SEQ ID NO: 5. 
     
     
         24 . The procoagulant protein according to  claim 18 , the monoclonal antibody or fragment thereof having an epitope, wherein the epitope comprises one or more residues selected from the group consisting of L36, P37, E38, G39, C40, Q41, P42, L43, V44, S45, S46, A47, V73, T74, L75, Q76, E77, E78, D79, A80, G81, E82, Y83, G84, C85, M86, R91, G92, P93, Q94, I95, L96, H97, R98, V99, S100, and L101 of SEQ ID NO: 5. 
     
     
         25 . The procoagulant protein according to  claim 18 , the monoclonal antibody or fragment thereof having an epitope, wherein the epitope comprises one or more residues selected from the group consisting of V17, Q18, C19, H20, Y21, R22, L23, Q24, D25, V26, K27, A28, R91, G92, P93, Q94, I95, L96, H97, R98, V99, S100, and L101 of SEQ ID NO: 5. 
     
     
         26 . The procoagulant protein according to  claim 18 , the monoclonal antibody or fragment thereof having an epitope, wherein the epitope comprises one or more residues selected from the group consisting of K118, 1119, G120, S121, L122, A123, E124, N125, A126, and F127 of SEQ ID NO: 6. 
     
     
         27 . The procoagulant protein according to  claim 18 , the monoclonal antibody or fragment thereof having an epitope, wherein the epitope comprises one or more residues selected from the group consisting of E5, T6, H7, K8, I9, G10, S11, L12, A13, E14, N15, A16, F17, S18, D19, and P20 of SEQ ID NO: 7. 
     
     
         28 . The procoagulant protein according to  claim 18 , the monoclonal antibody or fragment thereof having an epitope, wherein the epitope comprises one or more residues selected from the group consisting of K133, I134, G135, S136, L137, A138, N140, A141, F142, S143, D144, P145, and A146 of SEQ ID NO: 2. 
     
     
         29 . The procoagulant protein according to  claim 18 , the monoclonal antibody or fragment thereof having an epitope, wherein the epitope comprises K133, I134, G135, S136, L137, A138, N140, A141, F142, S143, D144, P145 and A146 of SEQ ID NO: 2. 
     
     
         30 . The procoagulant protein according to  claim 28 , wherein the heavy chain of the monoclonal antibody or fragment thereof comprises a CDR3 sequence of amino acids 117 to 121 (GVFTS) of SEQ ID NO: 40. 
     
     
         31 . The procoagulant protein according to  claim 28 , wherein the heavy chain of the monoclonal antibody or fragment thereof comprises:
 a CDR1 sequence of amino acids 49 to 53 (RYWMT) of SEQ ID NO: 40, wherein one of these amino acids may be substituted by a different amino acid; and   a CDR2 sequence of amino acids 68 to 84 (EINPDSSTINYNPSLKD) of SEQ ID NO: 40, wherein one, two, three or four of these amino acids may be substituted by a different amino acid; and/or   a CDR3 sequence of amino acids 117 to 121 (GVFTS) of SEQ ID NO: 40, wherein one, two or three of these amino acids may be substituted by a different amino acid;   
       and wherein the light chain of the monoclonal antibody or a fragment thereof comprises:
 a CDR1 sequence of amino acids 43 to 58 (RSSQSLVHRNGNTYFH) of SEQ ID NO: 41, wherein one, two, three or four of these amino acids may be substituted with a different amino acid; and/or 
 a CDR2 sequence of amino acids 74 to 80 (KVSNRFS) of SEQ ID NO: 41, wherein one or two of these amino acids may be substituted with a different amino acid; and/or 
 a CDR3 sequence of amino acids 113 to 121 (SQSTHVPYT) of SEQ ID NO: 41, wherein one or two of these amino acids may be substituted with a different amino acid. 
 
     
     
         32 . The procoagulant protein according to  claim 18 , which is a conjugate of the coagulation factor and the monoclonal antibody or a fragment thereof. 
     
     
         33 . A process for preparing the conjugate of  claim 32 , comprising:
 chemically conjugating the TLT-1 monoclonal antibody or fragment thereof, with one reactive group (RS1) of a linker and   reacting the coagulation factor with another reactive group (RS2) of said linker.   
     
     
         34 . The process of  claim 33 , wherein said linker is a polymer.