Multi-specific antigen-binding molecules and uses thereof

Abstract

Various bispecific antibodies that specifically bind to both blood coagulation factor IX/activated blood coagulation factor IX and blood coagulation factor X and functionally substitute for the cofactor function of blood coagulation factor VIII, that is, the function to promote activation of blood coagulation factor X by activated blood coagulation factor IX, were produced. From these antibodies, multispecific antigen-binding molecules having a high activity of functionally substituting for blood coagulation factor VIII were successfully discovered.

1 - 29 . (canceled) 
     
     
         30 . A bispecific antibody that recognizes blood coagulation factor IX and/or activated blood coagulation factor IX, and recognizes blood coagulation factor X, wherein the bispecific antibody comprises:
 a first antibody H chain comprising CDRs 1, 2, and 3 that comprise SEQ ID NOs: 105-107, respectively;   a second antibody H chain comprising CDRs 1, 2, and 3 that comprise:
 SEQ ID NOs: 126-128, respectively, 
 SEQ ID NOs: 129-131, respectively, or 
 SEQ ID NOs: 135-137, respectively; 
   a first antibody L chain comprising CDRs 1, 2, and 3 that comprise:
 SEQ ID NOs: 144-146, respectively, 
 SEQ ID NOs: 150-152, respectively, 
 SEQ ID NOs: 153-155, respectively, or 
 SEQ ID NOs: 156-158, respectively; and 
   a second antibody L chain comprising CDRs 1, 2, and 3 that comprise:
 SEQ ID NOs: 144-146, respectively, 
 SEQ ID NOs: 150-152, respectively, 
 SEQ ID NOs: 153-155, respectively, or 
 SEQ ID NOs: 156-158, respectively. 
   
     
     
         31 . The bispecific antibody of  claim 30 , wherein:
 the first antibody H chain comprises a variable region comprising SEQ ID NO: 45;   the second antibody H chain comprises a variable region comprising SEQ ID NO: 52, 53, or 55; and   each of the first and second antibody L chains comprises a variable region comprising a sequence independently selected from SEQ ID NOs: 58, 60, 61, and 62.   
     
     
         32 . The bispecific antibody of  claim 31 , wherein:
 the first antibody H chain comprises an antibody H chain constant region consisting of SEQ ID NO: 69, 70, 72, or 73;   the second antibody H chain comprises an antibody H chain constant region of:
 SEQ ID NO: 69 or 70, when the first antibody H chain comprises an antibody H chain constant region of SEQ ID NO: 72 or 73, or 
 SEQ ID NO: 72 or 73, when the first antibody H chain comprises an antibody H chain constant region of SEQ ID NO: 69 or 70; and 
   each of the first and second antibody L chains comprises an antibody L chain constant region consisting of SEQ ID NO: 74.   
     
     
         33 . The bispecific antibody of  claim 32 , wherein:
 the first antibody H chain comprises SEQ ID NO: 19 or 20;   the second antibody H chain comprises SEQ ID NO: 24, 25, 26, or 27; and   each of the first and second antibody L chains comprises a sequence independently selected from SEQ ID NO: 28, 30, 31, and 32.   
     
     
         34 . A bispecific antibody that recognizes blood coagulation factor IX and/or activated blood coagulation factor IX, and recognizes blood coagulation factor X, wherein the bispecific antibody comprises:
 a first antibody H chain that is an H chain of an antibody that binds to an epitope overlapping with an epitope bound by an antibody consisting of an antibody H chain consisting of SEQ ID NO: 19 or 20 and an antibody L chain consisting of SEQ ID NO: 28, 30, 31, or 32;   a second antibody H chain that is an H chain of an antibody that binds to an epitope overlapping with an epitope bound by an antibody consisting of an antibody H chain consisting of SEQ ID NO: 24, 25, 26, or 27 and an antibody L chain consisting of SEQ ID NO: 28, 30, 31, or 32; and   a first antibody L chain and a second antibody L chain, each of which is independently selected from (a) and (b):   (a) an L chain of an antibody that binds to an epitope overlapping with an epitope bound by an antibody that consists of an antibody H chain consisting of SEQ ID NO: 19 or 20 and an antibody L chain consisting of SEQ ID NO: 28, 30, 31, or 32; and   (b) an L chain of an antibody that binds to an epitope overlapping with an epitope bound by an antibody that consists of an antibody H chain consisting of SEQ ID NO: 24, 25, 26, or 27 and an antibody L chain consisting of SEQ ID NO: 28, 30, 31, or 32.   
     
     
         35 . The bispecific antibody of  claim 34 , wherein:
 the first antibody H chain comprises, by Kabat numbering, any one or more of an isoleucine at amino acid position 34; an asparagine, glutamine, or serine at amino acid position 35; a serine at amino acid position 49; an arginine at amino acid position 61; a glutamic acid at amino acid position 62; a serine or threonine at amino acid position 96; a lysine or arginine at amino acid position 98; a phenylalanine or tyrosine at amino acid position 100; a glycine at amino acid position 100b; and a tyrosine at amino acid position 102;   the second antibody H chain comprises, by Kabat numbering, any one or more of an aspartic acid at amino acid position 35; an arginine at amino acid position 53; a lysine at amino acid position 73; a glycine at amino acid position 76; a lysine or arginine at amino acid position 96; a tyrosine at amino acid position 98; a tyrosine at amino acid position 100; and a histidine at amino acid position 100a; and   the first antibody L chain and the second antibody L chain each independently comprises, by Kabat numbering, any one of more of a lysine or arginine at amino acid position 27; a glutamic acid at amino acid position 30; an arginine at amino acid position 31; a glutamine at amino acid position 32; an arginine or glutamine at amino acid position 50; a serine at amino acid position 52; an arginine at amino acid position 53; a lysine at amino acid position 54; a glutamic acid at amino acid position 55; a serine at amino acid position 92; a serine at amino acid position 93; a proline at amino acid position 94; and a proline at amino acid position 95.   
     
     
         36 . The bispecific antibody of  claim 30 , wherein the first and second antibody L chains are identical. 
     
     
         37 . The bispecific antibody of  claim 31 , wherein the first and second antibody L chains are identical. 
     
     
         38 . The bispecific antibody of  claim 32 , wherein the first and second antibody L chains are identical. 
     
     
         39 . The bispecific antibody of  claim 33 , wherein the first and second antibody L chains are identical. 
     
     
         40 . The bispecific antibody of  claim 34 , wherein the first and second antibody L chains are identical. 
     
     
         41 . The bispecific antibody of  claim 35 , wherein the first and second antibody L chains are identical. 
     
     
         42 . A bispecific antibody comprising a first polypeptide, a second polypeptide, a third polypeptide, and a fourth polypeptide, wherein:
 (a) the first polypeptide is an antibody H chain consisting of SEQ ID NO: 19, the second polypeptide is an antibody H chain consisting of SEQ ID NO: 24, and the third and fourth polypeptides are antibody L chains, each consisting of SEQ ID NO: 30;   (b) the first polypeptide is an antibody H chain consisting of SEQ ID NO: 19, the second polypeptide is an antibody H chain consisting of SEQ ID NO: 24, and the third and fourth polypeptides are antibody L chains, each consisting of SEQ ID NO: 31;   (c) the first polypeptide is an antibody H chain consisting of SEQ ID NO: 19, the second polypeptide is an antibody H chain consisting of SEQ ID NO: 27, and the third and fourth polypeptides are antibody L chains, each consisting of SEQ ID NO: 28;   (d) the first polypeptide is an antibody H chain consisting of SEQ ID NO: 20, the second polypeptide is an antibody H chain consisting of SEQ ID NO: 25, and the third and fourth polypeptides are antibody L chains, each consisting of SEQ ID NO: 32; or   (e) the first polypeptide is an antibody H chain consisting of SEQ ID NO: 20, the second polypeptide is an antibody H chain consisting of SEQ ID NO: 26, and the third and fourth polypeptides are antibody L chains, each consisting of SEQ ID NO: 31.   
     
     
         43 . A pharmaceutical composition comprising the bispecific antibody of  claim 30  and a pharmaceutically acceptable carrier. 
     
     
         44 . A pharmaceutical composition comprising the bispecific antibody of  claim 31  and a pharmaceutically acceptable carrier. 
     
     
         45 . A pharmaceutical composition comprising the bispecific antibody of  claim 34  and a pharmaceutically acceptable carrier. 
     
     
         46 . A pharmaceutical composition comprising the bispecific antibody of  claim 35  and a pharmaceutically acceptable carrier. 
     
     
         47 . A pharmaceutical composition comprising the bispecific antibody of  claim 36  and a pharmaceutically acceptable carrier. 
     
     
         48 . A pharmaceutical composition comprising the bispecific antibody of  claim 42  and a pharmaceutically acceptable carrier. 
     
     
         49 . A kit comprising the bispecific antibody of  claim 30 . 
     
     
         50 . A kit comprising the bispecific antibody of  claim 31 . 
     
     
         51 . A kit comprising the bispecific antibody of  claim 34 . 
     
     
         52 . A kit comprising the bispecific antibody of  claim 36 . 
     
     
         53 . A kit comprising the bispecific antibody of  claim 42 . 
     
     
         54 . A method for treating or reducing the incidence of bleeding, a disease accompanying bleeding, or a disease caused by bleeding in a subject in need thereof, the method comprising administering a therapeutically effective amount of the pharmaceutical composition of  claim 43  to the subject. 
     
     
         55 . A method for treating or reducing the incidence of bleeding, a disease accompanying bleeding, or a disease caused by bleeding in a subject in need thereof, the method comprising administering a therapeutically effective amount of the pharmaceutical composition of  claim 44  to the subject. 
     
     
         56 . A method for treating or reducing the incidence of bleeding, a disease accompanying bleeding, or a disease caused by bleeding in a subject in need thereof, the method comprising administering a therapeutically effective amount of the pharmaceutical composition of  claim 45  to the subject. 
     
     
         57 . A method for treating or reducing the incidence of bleeding, a disease accompanying bleeding, or a disease caused by bleeding in a subject in need thereof, the method comprising administering a therapeutically effective amount of the pharmaceutical composition of  claim 46  to the subject. 
     
     
         58 . A method for treating or reducing the incidence of bleeding, a disease accompanying bleeding, or a disease caused by bleeding in a subject in need thereof, the method comprising administering a therapeutically effective amount of the pharmaceutical composition of  claim 47  to the subject. 
     
     
         59 . A method for treating or reducing the incidence of bleeding, a disease accompanying bleeding, or a disease caused by bleeding in a subject in need thereof, the method comprising administering a therapeutically effective amount of the pharmaceutical composition of  claim 48  to the subject. 
     
     
         60 . A kit comprising the bispecific antibody of  claim 35 .