Anti-ALK2 antibody
This application provides: an antibody which specifically binds to an ALK2 protein and has an activity of inhibiting BMP signal transduction mediated by ALK2; a method for producing the antibody; and a pharmaceutical composition comprising the antibody, for treating and/or preventing ectopic ossification and/or bone dysplasia, anemia, or diffuse intrinsic pontine glioma (DIPG).
1. An antibody or an antigen-binding fragment thereof, comprising:
(i) heavy chain CDRs consisting of the amino acid sequences of SEQ ID NO: 65 for CDRH1, SEQ ID NO: 66 for CDRH2, and SEQ ID NO: 67 for CDRH3; and light chain CDRs consisting of the amino acid sequences of SEQ ID NO: 68 for CDRL1, SEQ ID NO: 69 for CDRL2, and SEQ ID NO: 70 for CDRL3;
(ii) a heavy chain variable region sequence consisting of amino acid numbers 20 to 140 of the amino acid sequence of SEQ ID NO: 107; and a light chain comprising a light chain variable region sequence consisting of amino acid numbers 21 to 129 of the amino acid sequence of SEQ ID NO: 52;
(iii) a heavy chain comprising the amino acid sequence consisting of amino acid numbers 20 to 470 of the amino acid sequence of SEQ ID NO: 107; and a light chain comprising the amino acid sequence consisting of amino acid numbers 21 to 234 of the amino acid sequence of SEQ ID NO: 52;
(iv) heavy chain CDRs consisting of the amino acid sequences of SEQ ID NO: 72 for CDRH1, SEQ ID NO: 73 for CDRH2, and SEQ ID NO: 74 for CDRH3; and light chain CDRs consisting of the amino acid sequences of SEQ ID NO: 75 for CDRL1, SEQ ID NO: 76 for CDRL2, and SEQ ID NO: 77 for CDRL3;
(v) a heavy chain variable region sequence consisting of amino acid numbers 20 to 137 of the amino acid sequence of SEQ ID NO: 113; and a light chain comprising a light chain variable region sequence consisting of amino acid numbers 21 to 128 of the amino acid sequence of SEQ ID NO: 119;
(vi) a heavy chain comprising the amino acid sequence consisting of amino acid numbers 20 to 467 of the amino acid sequence of SEQ ID NO: 113; and a light chain comprising the amino acid sequence consisting of amino acid numbers 21 to 233 of the amino acid sequence of SEQ ID NO: 119;
(vii) heavy chain CDRs consisting of the amino acid sequences of SEQ ID NO: 59 for CDRH1, SEQ ID NO: 60 for CDRH2, and SEQ ID NO: 61 for CDRH3; and light chain CDRs consisting of the amino acid sequences of SEQ ID NO: 62 for CDRL1, SEQ ID NO: 63 or 71 for CDRL2, and SEQ ID NO: 64 for CDRL3;
(viii) a heavy chain variable region sequence consisting of amino acid numbers 20 to 142 of the amino acid sequence of SEQ ID NO: 105; and a light chain comprising a light chain variable region sequence consisting of amino acid numbers 21 to 133 of the amino acid sequence of SEQ ID NO: 36;
(ix) a heavy chain comprising the amino acid sequence consisting of amino acid numbers 20 to 468 of the amino acid sequence of SEQ ID NO: 105; and a light chain comprising the amino acid sequence consisting of amino acid numbers 21 to 238 of the amino acid sequence of SEQ ID NO: 36;
(x) heavy chain CDRs consisting of the amino acid sequences of SEQ ID NO: 78 for CDRH1, SEQ ID NO: 79 for CDRH2, and SEQ ID NO: 80 for CDRH3; and light chain CDRs consisting of the amino acid sequences of SEQ ID NO: 81 for CDRL1, SEQ ID NO: 82 for CDRL2, and SEQ ID NO: 83 for CDRL3;
(xi) a heavy chain variable region sequence consisting of amino acid numbers 20 to 137 of the amino acid sequence of SEQ ID NO: 123; and a light chain comprising a light chain variable region sequence consisting of amino acid numbers 21 to 129 of the amino acid sequence of SEQ ID NO: 129; or
(xii) a heavy chain comprising the amino acid sequence consisting of amino acid numbers 20 to 467 of the amino acid sequence of SEQ ID NO: 123; and a light chain comprising the amino acid sequence consisting of amino acid numbers 21 to 234 of the amino acid sequence of SEQ ID NO: 129.
2. The antibody or antigen-binding fragment thereof according to claim 1 , comprising:
(i) heavy chain CDRs consisting of the amino acid sequences of SEQ ID NO: 65 for CDRH1, SEQ ID NO: 66 for CDRH2, and SEQ ID NO: 67 for CDRH3; and light chain CDRs consisting of the amino acid sequences of SEQ ID NO: 68 for CDRL1, SEQ ID NO: 69 for CDRL2, and SEQ ID NO: 70 for CDRL3;
(ii) a heavy chain variable region sequence consisting of amino acid numbers 20 to 140 of the amino acid sequence of SEQ ID NO: 107; and a light chain comprising a light chain variable region sequence consisting of amino acid numbers 21 to 129 of the amino acid sequence of SEQ ID NO: 52; or
(iii) a heavy chain comprising the amino acid sequence consisting of amino acid numbers 20 to 470 of the amino acid sequence of SEQ ID NO: 107; and a light chain comprising the amino acid sequence consisting of amino acid numbers 21 to 234 of the amino acid sequence of SEQ ID NO: 52.
3. The antibody or antigen-binding fragment thereof according to claim 2 , comprising heavy chain CDRs consisting of the amino acid sequences of SEQ ID NO: 65 for CDRH1, SEQ ID NO: 66 for CDRH2, and SEQ ID NO: 67 for CDRH3; and light chain CDRs consisting of the amino acid sequences of SEQ ID NO: 68 for CDRL1, SEQ ID NO: 69 for CDRL2, and SEQ ID NO: 70 for CDRL3.
4. The antibody or antigen-binding fragment thereof according to claim 2 , comprising a heavy chain variable region sequence consisting of amino acid numbers 20 to 140 of the amino acid sequence of SEQ ID NO: 107; and a light chain comprising a light chain variable region sequence consisting of amino acid numbers 21 to 129 of the amino acid sequence of SEQ ID NO: 52.
5. The antibody or antigen-binding fragment thereof according to claim 2 , comprising a heavy chain comprising the amino acid sequence consisting of amino acid numbers 20 to 470 of the amino acid sequence of SEQ ID NO: 107; and a light chain comprising the amino acid sequence consisting of amino acid numbers 21 to 234 of the amino acid sequence of SEQ ID NO: 52.
6. The antibody or antigen-binding fragment thereof according to claim 1 , comprising:
(i) heavy chain CDRs consisting of the amino acid sequences of SEQ ID NO: 72 for CDRH1, SEQ ID NO: 73 for CDRH2, and SEQ ID NO: 74 for CDRH3; and light chain CDRs consisting of the amino acid sequences of SEQ ID NO: 75 for CDRL1, SEQ ID NO: 76 for CDRL2, and SEQ ID NO: 77 for CDRL3;
(ii) a heavy chain variable region sequence consisting of amino acid numbers 20 to 137 of the amino acid sequence of SEQ ID NO: 113; and a light chain comprising a light chain variable region sequence consisting of amino acid numbers 21 to 128 of the amino acid sequence of SEQ ID NO: 119; or
(iii) a heavy chain comprising the amino acid sequence consisting of amino acid numbers 20 to 467 of the amino acid sequence of SEQ ID NO: 113; and a light chain comprising the amino acid sequence consisting of amino acid numbers 21 to 233 of the amino acid sequence of SEQ ID NO: 119.
7. The antibody or antigen-binding fragment thereof according to claim 6 , comprising heavy chain CDRs consisting of the amino acid sequences of SEQ ID NO: 72 for CDRH1, SEQ ID NO: 73 for CDRH2, and SEQ ID NO: 74 for CDRH3; and light chain CDRs consisting of the amino acid sequences of SEQ ID NO: 75 for CDRL1, SEQ ID NO: 76 for CDRL2, and SEQ ID NO: 77 for CDRL3.
8. The antibody or antigen-binding fragment thereof according to claim 6 , comprising a heavy chain variable region sequence consisting of amino acid numbers 20 to 137 of the amino acid sequence of SEQ ID NO: 113; and a light chain comprising a light chain variable region sequence consisting of amino acid numbers 21 to 128 of the amino acid sequence of SEQ ID NO: 119.
9. The antibody or antigen-binding fragment thereof according to claim 6 , comprising a heavy chain comprising the amino acid sequence consisting of amino acid numbers 20 to 467 of the amino acid sequence of SEQ ID NO: 113; and a light chain comprising the amino acid sequence consisting of amino acid numbers 21 to 233 of the amino acid sequence of SEQ ID NO: 119.
10. The antibody or an antigen-binding fragment thereof according to claim 1 , comprising:
(i) heavy chain CDRs consisting of the amino acid sequences of SEQ ID NO: 59 for CDRH1, SEQ ID NO: 60 for CDRH2, and SEQ ID NO: 61 for CDRH3; and light chain CDRs consisting of the amino acid sequences of SEQ ID NO: 62 for CDRL1, SEQ ID NO: 63 or 71 for CDRL2, and SEQ ID NO: 64 for CDRL3;
(ii) a heavy chain variable region sequence consisting of amino acid numbers 20 to 142 of the amino acid sequence of SEQ ID NO: 105; and a light chain comprising a light chain variable region sequence consisting of amino acid numbers 21 to 133 of the amino acid sequence of SEQ ID NO: 36; or
(iii) a heavy chain comprising the amino acid sequence consisting of amino acid numbers 20 to 468 of the amino acid sequence of SEQ ID NO: 105; and a light chain comprising the amino acid sequence consisting of amino acid numbers 21 to 238 of the amino acid sequence of SEQ ID NO: 36.
11. The antibody or antigen-binding fragment thereof according to claim 10 , comprising a heavy chain CDRs consisting of the amino acid sequences of SEQ ID NO: 59 for CDRH1, SEQ ID NO: 60 for CDRH2, and SEQ ID NO: 61 for CDRH3; and light chain CDRs consisting of the amino acid sequences of SEQ ID NO: 62 for CDRL1, SEQ ID NO: 63 or 71 for CDRL2, and SEQ ID NO: 64 for CDRL3.
12. The antibody or antigen-binding fragment thereof according to claim 10 , comprising a heavy chain variable region sequence consisting of amino acid numbers 20 to 142 of the amino acid sequence of SEQ ID NO: 105; and a light chain comprising a light chain variable region sequence consisting of amino acid numbers 21 to 133 of the amino acid sequence of SEQ ID NO: 36.
13. The antibody or antigen-binding fragment thereof according to claim 10 , comprising: a heavy chain comprising the amino acid sequence consisting of amino acid numbers 20 to 468 of the amino acid sequence of SEQ ID NO: 105; and a light chain comprising the amino acid sequence consisting of amino acid numbers 21 to 238 of the amino acid sequence of SEQ ID NO: 36.
14. An antibody or an antigen-binding fragment thereof according to claim 1 , comprising:
(i) heavy chain CDRs consisting of the amino acid sequences of SEQ ID NO: 78 for CDRH1, SEQ ID NO: 79 for CDRH2, and SEQ ID NO: 80 for CDRH3; and light chain CDRs consisting of the amino acid sequences of SEQ ID NO: 81 for CDRL1, SEQ ID NO: 82 for CDRL2, and SEQ ID NO: 83 for CDRL3;
(ii) a heavy chain variable region sequence consisting of amino acid numbers 20 to 137 of the amino acid sequence of SEQ ID NO: 123; and a light chain comprising a light chain variable region sequence consisting of amino acid numbers 21 to 129 of the amino acid sequence of SEQ ID NO: 129; or
(iii) a heavy chain comprising the amino acid sequence consisting of amino acid numbers 20 to 467 of the amino acid sequence of SEQ ID NO: 123; and a light chain comprising the amino acid sequence consisting of amino acid numbers 21 to 234 of the amino acid sequence of SEQ ID NO: 129.
15. The antibody or antigen-binding fragment thereof according to claim 14 , comprising heavy chain CDRs consisting of the amino acid sequences of SEQ ID NO: 78 for CDRH1, SEQ ID NO: 79 for CDRH2, and SEQ ID NO: 80 for CDRH3; and light chain CDRs consisting of the amino acid sequences of SEQ ID NO: 81 for CDRL1, SEQ ID NO: 82 for CDRL2, and SEQ ID NO: 83 for CDRL3.
16. The antibody or antigen-binding fragment thereof according to claim 14 , comprising a heavy chain variable region sequence consisting of amino acid numbers 20 to 137 of the amino acid sequence of SEQ ID NO: 123; and a light chain comprising a light chain variable region sequence consisting of amino acid numbers 21 to 129 of the amino acid sequence of SEQ ID NO: 129.
17. The antibody or antigen-binding fragment thereof according to claim 14 , comprising a heavy chain comprising the amino acid sequence consisting of amino acid numbers 20 to 467 of the amino acid sequence of SEQ ID NO: 123; and a light chain comprising the amino acid sequence consisting of amino acid numbers 21 to 234 of the amino acid sequence of SEQ ID NO: 129.
18. The antibody or an antigen-binding fragment thereof according to claim 1 , wherein the antibody or antigen binding fragment inhibits ALK2-mediated BMP signal transduction.
19. The antibody or an antigen-binding fragment thereof according to claim 1 , wherein the antibody binds to a wild-type ALK2 protein and mutant ALK2 protein, and optionally binds to an ALK2 extracellular region.
20. The antibody or the antigen-binding fragment thereof according to claim 1 , wherein the antibody or the antigen-binding fragment is a monoclonal antibody, a polyclonal antibody, a chimeric antibody, a humanized antibody, a human antibody, a single chain Fv, a bispecific antibody, or a multispecific antibody.
21. The antibody or the antigen-binding fragment thereof according to claim 1 , wherein the antibody comprises a heavy chain in which one to several carboxyl-terminal amino acids are deleted and/or has a pyroglutamylated amino-terminal amino acid residue in a heavy or light chain thereof.
22. A pharmaceutical composition comprising the antibody or antigen-binding fragment thereof according to claim 1 .
23. A pharmaceutical composition comprising the antibody or antigen-binding fragment thereof according to claim 2 .
24. A pharmaceutical composition comprising the antibody or antigen-binding fragment thereof according to claim 6 .
25. A pharmaceutical composition comprising the antibody or antigen-binding fragment thereof according to claim 10 .
26. A pharmaceutical composition comprising the antibody or antigen-binding fragment thereof according to claim 14 .
27. An antibody or an antigen binding fragment thereof, wherein the antibody comprises:
(a) a heavy chain comprising amino acids 20 to 142 of SEQ ID NO: 28 and a light chain comprising amino acids 21 to 133 of SEQ ID NO: 32;
(b) a heavy chain comprising amino acids 20 to 142 of SEQ ID NO: 28 and a light chain comprising amino acids 21 to 133 of SEQ ID NO: 34;
(c) a heavy chain comprising 20 to 142 of SEQ ID NO: 30 and a light chain comprising amino acids 21 to 133 of SEQ ID NO: 32;
(d) a heavy chain comprising 20 to 142 of SEQ ID NO: 30 and a light chain comprising amino acids 21 to 133 of SEQ ID NO: 34;
(e) a heavy chain comprising 20 to 142 of SEQ ID NO: 30 and a light chain comprising amino acids 21 to 133 of SEQ ID NO: 36;
(f) a heavy chain comprising 20 to 142 of SEQ ID NO: 30 and a light chain comprising amino acids 21 to 133 of SEQ ID NO: 38;
(g) a heavy chain comprising amino acids 20 to 142 of SEQ ID NO: 105 and a light chain comprising amino acids 21 to 133 of SEQ ID NO: 36;
(h) a heavy chain comprising amino acids 20 to 140 of SEQ ID NO: 40 and a light chain comprising amino acids 21 to 129 of SEQ ID NO: 50;
(i) a heavy chain comprising amino acids 20 to 140 of SEQ ID NO: 40 and a light chain comprising amino acids 21 to 129 of SEQ ID NO: 52;
(j) a heavy chain comprising amino acids 20 to 140 of SEQ ID NO: 40 and a light chain comprising amino acids 21 to 129 of SEQ ID NO: 54;
(k) a heavy chain comprising amino acids 20 to 140 of SEQ ID NO: 42 and a light chain comprising amino acids 21 to 129 of SEQ ID NO: 50;
(l) a heavy chain comprising amino acids 20 to 140 of SEQ ID NO: 42 and a light chain comprising amino acids 21 to 129 of SEQ ID NO: 52;
(m) a heavy chain comprising amino acids 20 to 140 of SEQ ID NO: 42 and a light chain comprising amino acids 21 to 129 of SEQ ID NO: 54;
(n) a heavy chain comprising amino acids 20 to 140 of SEQ ID NO: 44 and a light chain comprising amino acids 21 to 129 of SEQ ID NO: 50;
(o) a heavy chain comprising amino acids 20 to 140 of SEQ ID NO: 44 and a light chain comprising amino acids 21 to 129 of SEQ ID NO: 52;
(p) a heavy chain comprising amino acids 20 to 140 of SEQ ID NO: 44 and a light chain comprising amino acids 21 to 129 of SEQ ID NO: 54;
(q) a heavy chain comprising amino acids 20 to 140 of SEQ ID NO: 44 and a light chain comprising amino acids 21 to 129 of SEQ ID NO: 56;
(r) a heavy chain comprising amino acids 20 to 140 of SEQ ID NO: 46 and a light chain comprising amino acids 21 to 129 of SEQ ID NO: 54;
(s) a heavy chain comprising amino acids 20 to 140 of SEQ ID NO: 46 and a light chain comprising amino acids 21 to 129 of SEQ ID NO: 56;
(t) a heavy chain comprising amino acids 20 to 140 of SEQ ID NO: 46 and a light chain comprising amino acids 21 to 129 of SEQ ID NO: 58;
(u) a heavy chain comprising amino acids 20 to 140 of SEQ ID NO: 48 and a light chain comprising amino acids 21 to 129 of SEQ ID NO: 56;
(v) a heavy chain comprising amino acids 20 to 140 of SEQ ID NO: 107 and a light chain comprising amino acids 21 to 129 of SEQ ID NO: 52;
(w) a heavy chain comprising amino acids 20 to 140 of SEQ ID NO: 109 and a light chain comprising amino acids 21 to 129 of SEQ ID NO: 56;
(x) a heavy chain comprising amino acids 20 to 137 of SEQ ID NO: 111 and a light chain comprising amino acids 21 to 128 of SEQ ID NO: 115;
(y) a heavy chain comprising amino acids 20 to 137 of SEQ ID NO: 111 and a light chain comprising amino acids 21 to 128 of SEQ ID NO: 117;
(z) a heavy chain comprising amino acids 20 to 137 of SEQ ID NO: 111 and a light chain comprising amino acids 21 to 128 of SEQ ID NO: 119;
(aa) a heavy chain comprising amino acids 20 to 137 of SEQ ID NO: 113 and a light chain comprising amino acids 21 to 128 of SEQ ID NO: 115;
(bb) a heavy chain comprising amino acids 20 to 137 of SEQ ID NO: 113 and a light chain comprising amino acids 21 to 128 of SEQ ID NO: 117);
(cc) a heavy chain comprising amino acids 20 to 137 of SEQ ID NO: 113 and a light chain comprising amino acids 21 to 128 of SEQ ID NO: 119;
(dd) a heavy chain comprising amino acids 20 to 137 of SEQ ID NO: 121 and a light chain comprising amino acids 21 to 129 of SEQ ID NO: 125;
(ee) a heavy chain comprising amino acids 20 to 137 of SEQ ID NO: 121 and a light chain comprising amino acids 21 to 129 of SEQ ID NO: 127;
(ff) a heavy chain comprising amino acids 20 to 137 of SEQ ID NO: 121 and a light chain comprising amino acids 21 to 129 of SEQ ID NO: 129;
(gg) a heavy chain comprising amino acids 20 to 137 of SEQ ID NO: 123 and a light chain comprising amino acids 21 to 129 of SEQ ID NO: 125;
(hh) a heavy chain comprising amino acids 20 to 137 of SEQ ID NO: 123 and a light chain comprising amino acids 21 to 129 of SEQ ID NO: 127; or
(ii) a heavy chain comprising amino acids 20 to 137 of SEQ ID NO: 123 and a light chain comprising amino acids 21 to 129 of SEQ ID NO: 129.