Anti-gpiib/iiia antibodies or uses thereof

Abstract

The present invention provides antibodies and antigen-binding molecules thereof which specifically bind the α and/or β subunits of the non-active form of the GPIIb/IIIIa receptor. The antibodies and antigen-binding molecules can be genetically fused and/or conjugated to heterologous moieties and used, for example, as targeting moieties. The invention also includes methods for screening for these antibodies, as well as methods of making and methods of using chimeric molecules derived from the antibodies.

1 . An antibody or antigen-binding molecule thereof which specifically binds to a GPIIb/IIIa epitope,
 (a) wherein the antibody or antigen-binding molecule thereof specifically bind to the same GPIIb/IIIa epitope as an antibody selected from 34D10, 12B2, 2A2, 35D1, 36A8, 4B11, 1H6, 38G8, 21F10, 38A8, 18F7, 38F6, 13C1, 5C4, 23C10, 37C7, 28C2, 9D6, or 28F4;   (b) wherein the antibody or antigen-binding molecule thereof competitively inhibits GPIIb/IIIa binding by an antibody selected from 34D10, 12B2, 2A2, 35D1, 36A8, 4B11, 1H6, 38G8, 21F10, 38A8, 18F7, 38F6, 13C1, 5C4, 23C10, 37C7, 28C2, 9D6, or 28F4; or   (c) wherein the antibody or antigen-binding molecule thereof comprises at least one, at least two, at least three, at least four, or at least five complementarity determining regions (CDR) or variants thereof of an antibody selected from the 34D10, 12B2, 2A2, 35D1, 36A8, 4B11, 1H6, 38G8, 21F10, 38A8, 18F7, 38F6, 13C1, 5C4, 23C10, 37C7, 28C2, 9D6, or 28F4 antibodies.   
     
     
         2 . The antibody or antigen-binding molecule thereof of  claim 1 , which comprises six CDRs or variants thereof of an antibody selected from the 34D10, 12B2, 2A2, 35D1, 36A8, 4B11, 1H6, 38G8, 21F10, 38A8, 18F7, 38F6, 13C1, 5C4, 23C10, 37C7, 28C2, 9D6, or 28F4 antibodies. 
     
     
         3 . (canceled) 
     
     
         4 . An antibody or antigen-binding molecule thereof which specifically binds to a GPIIb/IIIa epitope, comprising
 (i) a VH-CDR1 comprising the consensus sequence XIYAMS wherein X 1  represents amino acid residues Thr (T), Ser (S), or Ala (A);   (ii) a VH-CDR2 comprising the consensus sequence SIX 2 X 3 GX 4 X 5 TYX 6 X 7 DSVKX 8  wherein X 2  represents amino acid residues Ser (S) or Asn (N), X 3  represents amino acid residues Ser (S) or Gly (G), X 4  represents amino acid residues Ser (S) or Gly (G), X 5  represents amino acid residues Ser (S), Asn (N), or Thr (T), X 6  represents amino acid residues Tyr (Y) or Phe (F), X 7  represents amino acid residues Leu (L) or Pro (P), and X 8  represents amino acids Gly (G) or Arg (R);   (iii) a VH-CDR3 comprising the consensus sequence GGDYGYAX 9 DY, wherein X 9  represents amino acid residues Leu (L) or Met (M);   (iv) a VL-CDR1 comprising the sequence RASSSVNYMY (SEQ ID NO: 28);   (v) a VL-CDR2 comprising the sequence YTSNLAP (SEQ ID NO: 29); and,   (vi) a VL-CDR3 comprising the sequence QQFSSSPWT (SEQ ID NO: 30).   
     
     
         5 . The antibody or antigen-binding molecule thereof according to  claim 4 , comprising
 (i) a VH-CDR1 sequence selected from SEQ ID NOS: 25, 31, 37, 43, or 111;   (ii) a VH-CDR2 sequence selected from SEQ ID NOS: 26, 32, 38, 44, or 112;   (iii) a VH-CDR3 sequence selected from SEQ ID NOS: 27, 33, 39, 45, or 113;   (iv) a VL-CDR1 sequence selected from SEQ ID NOS: 28, 34, 40, 117, or 114;   (v) a VL-CDR2 sequence selected from f SEQ ID NOS: 29, 35, 41, 118, or 115; and,   (vi) a VL-CDR3 sequence selected from SEQ ID NOS: 30, 36, 42, 119, or 116.   
     
     
         6 - 53 . (canceled) 
     
     
         54 . The antibody or antigen-binding molecule thereof of  claim 1 , wherein the antibody or antigen-binding molecule thereof comprises or consists of:
 (a) a single chain Fv (“scFv”);   (b) a diabody;   (c) a minibody;   (d) a polypeptide chain of an antibody;   (e) F(ab′) 2 ; or,   (f) F(ab).   
     
     
         55 . A chimeric molecule comprising,
 (i) the antibody or antigen-binding molecule thereof of  claim 1 ;   (ii) a heterologous moiety; and,   (iii) an optional linker.   
     
     
         56 . (canceled) 
     
     
         57 . The chimeric molecule according to  claim 55 , wherein the heterologous moiety comprises a clotting factor. 
     
     
         58 . The chimeric molecule according to  claim 57 , wherein the clotting factor is independently selected from the group consisting of FVII, FIX, FX, and any combinations thereof. 
     
     
         59 . The chimeric molecule according to  claim 58 , wherein the clotting factor is FVII zymogen, activatable FVII, activated FVII (FVIIa), FIX zymogen, activatable FIX, activated FIX (FIXa), FX zymogen, activatable FX, or activated FX (FXa). 
     
     
         60 . The chimeric molecule according to  claim 55 , further comprising a second heterologous moiety. 
     
     
         61 . The chimeric molecule according to  claim 60 , which comprises a formula selected from:
 (i) H1-(L1)-Tm-(L2)-H2;   (ii) H2-(L2)-Tm-(L1)-H1;   (iii) H1-(L1)-H2-(L2)-Tm;   (iv) H2-(L2)-H1-(L1)-Tm;   (v) Tm-(L1)-H1-(L2)-H2; or,   (vi) Tm-(L2)-H2-(L1)-H1   wherein,   Tm is the antibody or antigen-binding molecule thereof of  claim 1 ;   H1 is the first heterologous moiety,   H2 is the second heterologous moiety,   L1 is the first optional linker,   L2 is the second optional linker.   
     
     
         62 . (canceled) 
     
     
         63 . The chimeric molecule according to  claim 60 , wherein the second heterologous moiety comprise a half-life extending moiety. 
     
     
         64 .- 68 . (canceled) 
     
     
         69 . A chimeric protein comprising a first polypeptide chain and a second polypeptide chain, which are associated with each other,
 (1) wherein the first polypeptide chain comprises a light chain of a clotting factor and a heterologous moiety, and the second polypeptide chain comprises a heavy chain of the clotting factor and the antibody or antigen-binding molecule thereof of  claim 1 ;   (2) wherein the first polypeptide chain comprises a light chain of a clotting factor and the antibody or antigen-binding molecule thereof of  claim 1  and the second polypeptide chain comprises a heavy chain of the clotting factor and a heterologous moiety;   (3) wherein the first polypeptide chain comprises a light chain of a clotting factor and the second polypeptide chain comprises a heavy chain of the clotting factor, the antibody or antigen-binding molecule thereof of  claim 1 , and a heterologous moiety;   (4) wherein the first polypeptide chain comprises a light chain of a clotting factor and the second polypeptide chain comprises a heavy chain of the clotting factor, a heterologous moiety, and the antibody or antigen-binding molecule thereof of  claim 1 ;   (5) wherein the first polypeptide chain comprises a light chain of a clotting factor, a heterologous moiety, and the antibody or antigen-binding molecule thereof of  claim 1  and the second polypeptide chain comprises a heavy chain of the clotting factor; or   (6) wherein the first polypeptide chain comprises a light chain of a clotting factor, the antibody or antigen-binding molecule thereof of  claim 1 , and a heterologous moiety and the second polypeptide chain comprises a heavy chain of the clotting factor.   
     
     
         70 - 76 . (canceled) 
     
     
         77 . The chimeric molecule according to  claim 69 , wherein the heterologous moiety comprise a half-life extending moiety comprising a low-complexity polypeptide. 
     
     
         78 - 81 . (canceled) 
     
     
         82 . A chimeric molecule comprising a first polypeptide chain and a second polypeptide chain, which are associated with each other,
 (1) wherein the first polypeptide chain comprises a light chain of a clotting factor and a targeting moiety, which binds to a platelet, and the second polypeptide chain comprises a heavy chain of the clotting factor and a heterologous moiety;   (2) wherein the first polypeptide chain comprises a light chain of a clotting factor and a heterologous moiety and the second polypeptide chain comprises a heavy chain of the clotting factor and a targeting moiety, which binds to a platelet;   (3) wherein the first polypeptide chain comprises a light chain of a clotting factor, a heterologous moiety, and a targeting moiety, which binds to a platelet, and the second polypeptide comprises a heavy chain of the clotting factor; or   (4) wherein the first polypeptide chain comprises a light chain of a clotting factor, a targeting moiety, which binds to a platelet, and a heterologous moiety and the second polypeptide chain comprises a heavy chain of the clotting factor, and   wherein the clotting factor is FVII, FIX, or FX.   
     
     
         83 . The chimeric molecule of  claim 82 ,
 (1) wherein the first polypeptide chain comprises CF L -L1-H or H-L1-CF L  and the second polypeptide chain comprises CF H -L2-Tm or Tm-L2-CF H ;   (2) wherein the first polypeptide chain comprises CF L -L1-Tm or Tm-L1-CF L  and the second polypeptide chain comprises CF H -L2-H or H-L2-CF H ;   (3) wherein the first polypeptide chain comprises CF L -L1-H-L2-Tm or Tm-L1-H-L2-CF L  and the second polypeptide chain comprises CF H ; or   (4) wherein the first polypeptide chain comprises CF L -L1-Tm-L2-H or H-L1-Tm-L2-CF L  and the second polypeptide chain comprises CF H ;   wherein   H is the heterologous moiety;   CF H  is the heavy chain of the clotting factor;   CF L  is the light chain of the clotting factor;   Tm is the targeting moiety, which binds to a platelet;   L1 is a first optional linker; and   L2 is a second optional linker.   
     
     
         84 - 99 . (canceled) 
     
     
         100 . A nucleic acid molecule or a set of nucleic acid molecules encoding an antibody or antigen-binding molecule thereof of  claim 1  or a complement thereof. 
     
     
         101 . A vector or a set of vectors comprising the nucleic acid molecule or the set of the nucleic acid molecules of  claim 100  or a complement thereof. 
     
     
         102 . A host cell comprising the vector according to  claim 101 . 
     
     
         103 . A method for producing an antibody or antigen-binding molecule thereof comprising culturing the host cell according to  claim 102  and recovering the antibody or antigen-binding molecule thereof from the culture medium. 
     
     
         104 . A pharmaceutical composition comprising the antibody or antigen-binding molecule thereof of  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         105 . A method of reducing a frequency or degree of a bleeding episode in a subject in need thereof comprising administering the antibody or antigen-binding molecule thereof of  claim 1 . 
     
     
         106 - 108 . (canceled) 
     
     
         109 . The method according to  claim 105 , wherein the bleeding episode is caused by a blood coagulation disorder. 
     
     
         110 - 114 . (canceled) 
     
     
         115 . A method to target a therapeutic or prophylactic agent to the surface of platelets comprising fusing the agent to the antibody or antigen-binding molecule thereof according to  claim 1 . 
     
     
         116 . A method to increase the activity of a therapeutic or prophylactic agent comprising fusing the agent to the antibody or antigen-binding molecule thereof according to  claim 1 . 
     
     
         117 . (canceled) 
     
     
         118 . A method to improve the pharmacokinetic properties of a clotting factor comprising fusing the clotting factor to the antibody or antigen-binding molecule thereof according to  claim 1 . 
     
     
         119 . (canceled) 
     
     
         120 . A method of measuring the level of platelets in plasma of a subject in need thereof comprising contacting the antibody or antigen binding molecule thereof of  claim 1  with the plasma from the subject and measuring the plasma level.