Antibody-drug conjugate against receptor tyrosine kinase EphA5

Abstract

This provides a novel lung cancer therapy that targets tumor-specific targets such as EphA5. [Solution] The present invention provides antibodies, binding polypeptides, and immunoconjugates that are specific to human EPH receptor A5 (EphA5) in certain contexts.

Claims

1. An antibody or its antigen-binding fragment that specifically binds to the epitope of the human EPH receptor A5, and the following: a. A heavy chain variable region comprising three heavy chain complementarity determination regions (HCDRs), HCDR1 contains the amino acid sequence shown in SEQ ID NO: 16, HCDR2 contains an amino acid sequence selected from the group consisting of SEQ ID NO: 17 to 19, and HCDR3 has a heavy chain variable region containing the amino acid sequence shown in SEQ ID NO: 20; b. A light chain variable region comprising three light chain complementarity determination regions (LCDRs), LCDR1 contains an amino acid sequence selected from the group consisting of SEQ ID NO: 21 and 22. LCDR2 contains the amino acid sequence shown in SEQ ID NO: 23, and LCDR3 contains the amino acid sequence shown in SEQ ID NO: 24, and the light chain variable region Antibody or antigen-binding fragments containing such fragments.

2. a. A heavy chain containing a variable region that includes an amino acid sequence having at least 85% sequence identity with SEQ ID NO:1; b. Light chain and a variable region containing an amino acid sequence having at least 85% sequence identity with SEQ ID NO: 8 The antibody or antigen-binding fragment according to claim 1, further comprising:

3. a. The heavy chain variable region includes HCDR1 containing the amino acid sequence shown in SEQ ID NO: 16, HCDR2 containing the amino acid sequence shown in SEQ ID NO: 17, and HCDR3 containing the amino acid sequence shown in SEQ ID NO: 20; and the light chain variable region includes LCDR1 containing the amino acid sequence shown in SEQ ID NO: 21, LCDR2 containing the amino acid sequence shown in SEQ ID NO: 23, and LCDR3 containing the amino acid sequence shown in SEQ ID NO: 24; b. The heavy chain variable region includes HCDR1 containing the amino acid sequence shown in SEQ ID NO: 16, HCDR2 containing the amino acid sequence shown in SEQ ID NO: 18, and HCDR3 containing the amino acid sequence shown in SEQ ID NO: 20; and the light chain variable region includes LCDR1 containing the amino acid sequence shown in SEQ ID NO: 21, LCDR2 containing the amino acid sequence shown in SEQ ID NO: 23, and LCDR3 containing the amino acid sequence shown in SEQ ID NO: 24; c. The heavy chain variable region includes HCDR1 containing the amino acid sequence shown in SEQ ID NO: 16, HCDR2 containing the amino acid sequence shown in SEQ ID NO: 19, and HCDR3 containing the amino acid sequence shown in SEQ ID NO: 20; and the light chain variable region includes LCDR1 containing the amino acid sequence shown in SEQ ID NO: 21, LCDR2 containing the amino acid sequence shown in SEQ ID NO: 23, and LCDR3 containing the amino acid sequence shown in SEQ ID NO: 24; d. The heavy chain variable region includes HCDR1 containing the amino acid sequence shown in SEQ ID NO: 16, HCDR2 containing the amino acid sequence shown in SEQ ID NO: 17, and HCDR3 containing the amino acid sequence shown in SEQ ID NO: 20; and the light chain variable region includes LCDR1 containing the amino acid sequence shown in SEQ ID NO: 22, LCDR2 containing the amino acid sequence shown in SEQ ID NO: 23, and LCDR3 containing the amino acid sequence shown in SEQ ID NO: 24; e. The heavy chain variable region comprises HCDR1 containing the amino acid sequence shown in SEQ ID NO: 16, HCDR2 containing the amino acid sequence shown in SEQ ID NO: 18, and HCDR3 containing the amino acid sequence shown in SEQ ID NO: 20; and the light chain variable region comprises LCDR1 containing the amino acid sequence shown in SEQ ID NO: 22, LCDR2 containing the amino acid sequence shown in SEQ ID NO: 23, and LCDR3 containing the amino acid sequence shown in SEQ ID NO: 24; or f. The antibody or antigen-binding fragment according to claim 1 or 2, wherein the heavy chain variable region comprises HCDR1 containing the amino acid sequence shown in SEQ ID NO: 16, HCDR2 containing the amino acid sequence shown in SEQ ID NO: 19, and HCDR3 containing the amino acid sequence shown in SEQ ID NO: 20; and the light chain variable region comprises LCDR1 containing the amino acid sequence shown in SEQ ID NO: 22, LCDR2 containing the amino acid sequence shown in SEQ ID NO: 23, and LCDR3 containing the amino acid sequence shown in SEQ ID NO:

24.

4. The antibody or antigen-binding fragment according to any one of claims 1 to 3, wherein the amino acid sequence of the heavy chain variable region has at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with an amino acid sequence selected from the group consisting of SEQ ID NO: 2 to 7.

5. The antibody or antigen-binding fragment according to any one of claims 1 to 4, wherein the heavy chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 2 to 7.

6. The antibody or antigen-binding fragment according to any one of claims 1 to 5, wherein the heavy chain variable region consists of an amino acid sequence selected from the group consisting of SEQ ID NO: 2 to 7.

7. The antibody or antigen-binding fragment according to any one of claims 1 to 6, wherein the heavy chain variable region is encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 26 to 31.

8. The antibody or antigen-binding fragment according to any one of claims 1 to 7, wherein the amino acid sequence of the light chain variable region has at least 85%, 90%, 95%, 96%, 97%, 98%, or 99% sequence identity with an amino acid sequence selected from the group consisting of SEQ ID NO: 9 to 13.

9. The antibody or antigen-binding fragment according to any one of claims 1 to 3 and 8, wherein the light chain variable region comprises an amino acid sequence selected from the group consisting of SEQ ID NO: 9 to 13.

10. The antibody or antigen-binding fragment according to any one of claims 1 to 3 and 8 to 9, wherein the light chain variable region consists of an amino acid sequence selected from the group consisting of SEQ ID NO: 9 to 13.

11. The antibody or antigen-binding fragment according to any one of claims 1 to 3 and 8 to 9, wherein the light chain variable region is encoded by a nucleic acid sequence selected from the group consisting of SEQ ID NO: 33 to 37.

12. a. The heavy chain variable region is indicated by SEQ ID NO: 2, and the light chain variable region is indicated by SEQ ID NO: 9; b. The heavy chain variable region is indicated by SEQ ID NO: 2, and the light chain variable region is indicated by SEQ ID NO: 10; c. The heavy chain variable region is indicated by SEQ ID NO: 2, and the light chain variable region is indicated by SEQ ID NO: 11; d. Whether the heavy chain variable region is indicated by SEQ ID NO: 2 and the light chain variable region is indicated by SEQ ID NO: 12; e. The heavy chain variable region is indicated by SEQ ID NO: 2, and the light chain variable region is indicated by SEQ ID NO: 13; f. Whether the heavy chain variable region is indicated by SEQ ID NO: 3 and the light chain variable region is indicated by SEQ ID NO: 9; g. The heavy chain variable region is indicated by SEQ ID NO: 3, and the light chain variable region is indicated by SEQ ID NO: 10; h. Is the heavy chain variable region indicated by SEQ ID NO: 3, and the light chain variable region indicated by SEQ ID NO: 11? i. Is the heavy chain variable region indicated by SEQ ID NO: 3, and the light chain variable region indicated by SEQ ID NO: 12? j. Whether the heavy chain variable region is indicated by SEQ ID NO: 3 and the light chain variable region is indicated by SEQ ID NO: 13; k. The heavy chain variable region is indicated by SEQ ID NO: 4, and the light chain variable region is indicated by SEQ ID NO: 9; l. The heavy chain variable region is indicated by SEQ ID NO: 4, and the light chain variable region is indicated by SEQ ID NO: 10; m. The heavy chain variable region is indicated by SEQ ID NO: 4, and the light chain variable region is indicated by SEQ ID NO: 11; n. The heavy chain variable region is indicated by SEQ ID NO: 4, and the light chain variable region is indicated by SEQ ID NO: 12; o. The heavy chain variable region is indicated by SEQ ID NO: 4, and the light chain variable region is indicated by SEQ ID NO: 13; p. Is the heavy chain variable region indicated by SEQ ID NO: 5, and the light chain variable region indicated by SEQ ID NO: 9? q. Is the heavy chain variable region indicated by SEQ ID NO: 5, and the light chain variable region indicated by SEQ ID NO: 10? r. The heavy chain variable region is indicated by SEQ ID NO: 5, and the light chain variable region is indicated by SEQ ID NO: 11; s. The heavy chain variable region is indicated by SEQ ID NO: 5, and the light chain variable region is indicated by SEQ ID NO: 12; t. The heavy chain variable region is indicated by SEQ ID NO: 5, and the light chain variable region is indicated by SEQ ID NO: 13; u. The heavy chain variable region is indicated by SEQ ID NO: 6, and the light chain variable region is indicated by SEQ ID NO: 9; v. Whether the heavy chain variable region is indicated by SEQ ID NO: 6 and the light chain variable region is indicated by SEQ ID NO: 10; w. The heavy chain variable region is indicated by SEQ ID NO: 6, and the light chain variable region is indicated by SEQ ID NO: 11; x. The heavy chain variable region is indicated by SEQ ID NO: 6, and the light chain variable region is indicated by SEQ ID NO: 12; y. The heavy chain variable region is indicated by SEQ ID NO: 6, and the light chain variable region is indicated by SEQ ID NO: 13; z. The heavy chain variable region is indicated by SEQ ID NO: 7, and the light chain variable region is indicated by SEQ ID NO: 9; aa. The heavy chain variable region is indicated by SEQ ID NO: 7, and the light chain variable region is indicated by SEQ ID NO: 10; bb. The heavy chain variable region is indicated by SEQ ID NO: 7, and the light chain variable region is indicated by SEQ ID NO: 11; cc. The heavy chain variable region is shown in SEQ ID NO: 7, and the light chain variable region is shown in SEQ ID NO: 12; or dd. The antibody or antigen-binding fragment according to any one of claims 1 to 11, wherein the heavy chain variable region is indicated by SEQ ID NO: 7 and the light chain variable region is indicated by SEQ ID NO:

13.

13. a. The heavy chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 2, and the light chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 9; b. The heavy chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 2, and the light chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 10; c. The heavy chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 2, and the light chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 11; d. The heavy chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 2, and the light chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 12; e. The heavy chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 2, and the light chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 13; f. The heavy chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 3, and the light chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 9; g. The heavy chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 3, and the light chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 10; h. The heavy chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 3, and the light chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 11; i. The heavy chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 3, and the light chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 12; j. The heavy chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 3, and the light chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 13; k. The heavy chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 4, and the light chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 9; l. The heavy chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 4, and the light chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 10; m. The heavy chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 4, and the light chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 11; n. The heavy chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 4, and the light chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 12; o. The heavy chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 4, and the light chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 13; p. Whether the heavy chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 5, and whether the light chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 9; q. Does the heavy chain variable region contain a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 5, and does the light chain variable region contain a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 10? r. The heavy chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 5, and the light chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 11; s. The heavy chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 5, and the light chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 12; t. The heavy chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 5, and the light chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 13; u. The heavy chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 6, and the light chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 9; v. The heavy chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 6, and the light chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 10; w. The heavy chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 6, and the light chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 11; x. The heavy chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 6, and the light chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 12; y. The heavy chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 6, and the light chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 13; z. The heavy chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 7, and the light chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 9; aa. The heavy chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 7, and the light chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 10; bb. The heavy chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 7, and the light chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 11; cc. The heavy chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 7, and the light chain variable region contains a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 12; or dd. The antibody or antigen-binding fragment according to any one of claims 1 to 11, wherein the heavy chain variable region comprises a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO: 7, and the light chain variable region comprises a sequence having at least 90% sequence identity with the sequence shown in SEQ ID NO:

13.

14. a. The heavy chain includes a heavy chain complementarity determination region 1 (HCDR1) containing the sequence shown in SEQ ID NO: 16, a heavy chain variable region containing the sequence shown in SEQ ID NO: 18, and a heavy chain variable region containing the sequence shown in SEQ ID NO: 20; b. A light chain variable region comprising light chain complementarity determination region 1 (LCDR1) containing the sequence shown in SEQ ID NO: 21, LCDR2 containing the sequence shown in SEQ ID NO: 23a, and LCDR3 containing the sequence shown in SEQ ID NO: 24, and An antibody or its antigen-binding fragment that specifically binds to the human EPH receptor A5 epitope, including [the specified antibody].

15. a. The heavy chain includes a heavy chain complementarity determination region 1 (HCDR1) containing the sequence shown in SEQ ID NO: 16, a heavy chain variable region containing the sequence shown in SEQ ID NO: 19, and a heavy chain variable region containing the sequence shown in SEQ ID NO: 20; b. A light chain variable region comprising light chain complementarity determination region 1 (LCDR1) containing the sequence shown in SEQ ID NO: 21, LCDR2 containing the sequence shown in SEQ ID NO: 23, and LCDR3 containing the sequence shown in SEQ ID NO: 24, and An antibody or its antigen-binding fragment that specifically binds to the human EPH receptor A5 epitope, including [the specified antibody].

16. a. The heavy chain includes a heavy chain complementarity determination region 1 (HCDR1) containing the sequence shown in SEQ ID NO: 16, a heavy chain variable region containing the sequence shown in SEQ ID NO: 17, and a heavy chain variable region containing the sequence shown in SEQ ID NO: 20; b. A light chain variable region including complementarity determination region 1 (LCDR1) containing the sequence shown in SEQ ID NO: 22, LCDR2 containing the sequence shown in SEQ ID NO: 23, and LCDR3 containing the sequence shown in SEQ ID NO: 24, and An antibody or its antigen-binding fragment that specifically binds to the human EPH receptor A5 epitope, including [the specified antibody].

17. a. The heavy chain includes a heavy chain complementarity determination region 1 (HCDR1) containing the sequence shown in SEQ ID NO: 16, a heavy chain variable region containing the sequence shown in SEQ ID NO: 18, and a heavy chain variable region containing the sequence shown in SEQ ID NO: 20; b. A light chain variable region comprising light chain complementarity determination region 1 (LCDR1) containing the sequence shown in SEQ ID NO: 22, LCDR2 containing the sequence shown in SEQ ID NO: 23, and LCDR3 containing the sequence shown in SEQ ID NO: 24, and An antibody or its antigen-binding fragment that specifically binds to the human EPH receptor A5 epitope, including [the specified antibody].

18. a. A heavy chain containing a heavy chain variable region including the amino acid sequence shown in SEQ ID NO: 5; b. Light chain and a light chain variable region including the amino acid sequence shown in SEQ ID NO: 11 An antibody or its antigen-binding fragment, including an antibody.

19. a. A heavy chain containing a heavy chain variable region including the amino acid sequence shown in SEQ ID NO: 6; b. Light chain and a light chain variable region including the amino acid sequence shown in SEQ ID NO: 11 An antibody or its antigen-binding fragment, including an antibody.

20. An antibody or antigen-binding fragment thereof according to any one of claims 1 to 19, selected from the group consisting of a full-length antibody, a Fab, and a single-chain variable fragment (scFv).

21. An antibody or antigen-binding fragment thereof according to any one of claims 1 to 20, which is a full-length antibody.

22. A humanized antibody or antigen-binding fragment thereof according to any one of claims 1 to 21.

23. The antibody or antigen-binding fragment according to any one of claims 1 to 22, wherein the heavy chain further comprises a constant domain of a human immunoglobulin heavy chain, and the light chain further comprises a constant domain of a human light chain.

24. The antibody or antigen-binding fragment according to claim 23, wherein the constant domain of the human immunoglobulin heavy chain is derived from the IgG1 heavy chain.

25. The antibody or antigen-binding fragment according to claim 23 or 224, wherein the constant domain of the human immunoglobulin heavy chain comprises the amino acid sequence shown in SEQ ID NO:

14.

26. The antibody according to claim 23, wherein the constant domain of the human light chain is derived from the human kappa light chain.

27. The antibody or antigen-binding fragment according to claim 23 or claim 26, wherein the constant domain of the human light chain comprises the amino acid sequence shown in SEQ ID NO:

15.

28. a. A heavy chain containing at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with respect to the amino acid sequence shown in SEQ ID NO: 38; b. Light chain containing at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with respect to the amino acid sequence shown in SEQ ID NO:

40. An antibody or its antigen-binding fragment that specifically binds to the human EPH receptor A5 epitope, including [the specified antibody].

29. a. The variable region of the heavy chain includes HCDR1 containing the amino acid sequence shown in SEQ ID NO: 16, HCDR2 containing the amino acid sequence shown in SEQ ID NO: 17, and HCDR3 containing the amino acid sequence shown in SEQ ID NO: 20; and b. The antibody or antigen-binding fragment according to claim 28, wherein the variable region of the light chain comprises LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 21, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 23, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO:

24.

30. An antibody or antigen-binding fragment according to any one of claims 1 to 29, wherein the heavy chain is indicated by SEQ ID NO: 38 and the light chain is indicated by SEQ ID NO:

40.

31. a. A heavy chain containing at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with respect to the amino acid sequence shown in SEQ ID NO: 39; b. Light chain containing at least 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity with respect to the amino acid sequence shown in SEQ ID NO:

40. An antibody or its antigen-binding fragment that specifically binds to the human EPH receptor A5 epitope, including [the specified antibody].

32. a. The variable region of the heavy chain includes HCDR1 containing the amino acid sequence shown in SEQ ID NO: 16, HCDR2 containing the amino acid sequence shown in SEQ ID NO: 18, and HCDR3 containing the amino acid sequence shown in SEQ ID NO: 20; and b. The antibody or antigen-binding fragment according to claim 31, wherein the variable region of the light chain comprises LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 21, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 23, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO:

24.

33. An antibody or antigen-binding fragment according to any one of claims 1 to 27, 31, and 32, wherein the heavy chain is indicated by SEQ ID NO: 39 and the light chain is indicated by SEQ ID NO:

40.

34. Dissociation constant (K) for binding to human EpHA5 D ) is 1.25 × 10 -9 An antibody or antigen-binding fragment according to any one of claims 1 to 33, wherein the M value is less than M.

35. Dissociation constant (K) for binding to human EpHA5 D ) but 8×10 -10 M~1.1×10 -9 The antibody or antigen-binding fragment according to any one of claims 1 to 34, wherein M.

36. Regarding binding to human EpHA5, the dissociation constant (K) is within 2 times that of the reference antibody, indicating good binding. D An antibody or antigen-binding fragment having ) the antibody or antigen-binding fragment according to any one of claims 1 to 35, wherein the reference antibody is an antibody comprising a variable heavy chain indicated by 11C12 or SEQ ID NO: 1 and a variable light chain indicated by SEQ ID NO: 8, and optionally the reference antibody is of the same form.

37. Dissociation constant (K) for binding to human EpHA5 D An antibody or antigen-binding fragment thereof which is more than 1.2 times better than a reference antibody, wherein the reference antibody is an antibody comprising a variable heavy chain indicated by 11C12 or SEQ ID NO: 1 and a variable light chain indicated by SEQ ID NO: 8, and optionally the reference antibody is of the same form, according to any one of claims 1 to 36.

38. Dissociation constant (K D The antibody or antigen-binding fragment according to any one of claims 34 to 37, which is determined by Biacore.

39. An antibody or antigen-binding fragment thereof exhibiting increased thermal stability compared to a reference antibody, wherein the reference antibody is an antibody comprising a variable heavy chain indicated by 11C12 or SEQ ID NO: 1 and a variable light chain indicated by SEQ ID NO: 8, and optionally the reference antibody is of the same form, according to any one of claims 1 to 38.

40. An antibody or antigen-binding fragment thereof having a melting temperature profile having a Tm1 that is greater than 5°C or about 5°C greater than that of a reference antibody, wherein the reference antibody is an antibody comprising a variable heavy chain indicated by 11C12 or SEQ ID NO: 1 and a variable light chain indicated by SEQ ID NO: 8, and optionally the reference antibody is of the same form, according to any one of claims 1 to 39.

41. An antibody or antigen-binding fragment thereof having a melting temperature profile having a Tm1 that is greater than 10°C or about 10°C greater than that of a reference antibody, wherein the reference antibody is an antibody comprising a variable heavy chain indicated by 11C12 or SEQ ID NO: 1 and a variable light chain indicated by SEQ ID NO: 8, and optionally the reference antibody is of the same form, according to any one of claims 1 to 40.

42. The antibody or antigen-binding fragment according to claim 40 or 41, wherein the Tm1 of the thermal denaturation curve is greater than 60°C.

43. The antibody or antigen-binding fragment according to any one of claims 40 to 42, wherein the Tm1 is approximately 60°C to approximately 70°C.

44. The antibody or antigen-binding fragment according to any one of claims 40 to 43, wherein the Tm1 is approximately 64°C, approximately 65°C, approximately 66°C, approximately 67°C, approximately 68°C, or approximately 69°C.

45. The antibody or antigen-binding fragment according to any one of claims 40 to 44, wherein the melting temperature profile is monophasic.

46. An antibody or antigen-binding fragment thereof, wherein the aggregation onset temperature (Tagg) is increased by approximately 1°C, approximately 2°C, approximately 3°C, approximately 4°C, or approximately 5°C compared to a reference antibody, wherein the reference antibody is an antibody comprising a variable heavy chain indicated by 11C12 or SEQ ID NO: 1 and a variable light chain indicated by SEQ ID NO: 8, according to any one of claims 1 to 45.

47. The antibody or antigen-binding fragment according to any one of claims 1 to 46, wherein the aggregation onset temperature (Tagg) is greater than approximately 67°C.

48. The antibody or antigen-binding fragment according to any one of claims 1 to 47, wherein the aggregation onset temperature (Tagg) is approximately 67°C to approximately 71°C.

49. The antibody or antigen-binding fragment according to any one of claims 1 to 48, wherein the Tag is approximately 67°C, approximately 68°C, approximately 69°C, approximately 70°C, or approximately 71°C.

50. An antibody or antigen-binding fragment according to any one of claims 1 to 49, which binds to human EphA5 on the cell surface.

51. An antibody or antigen-binding fragment thereof that binds to human EphA5 expressed on the surface of cells, optionally on the surface of the H460 cell line, with an increased EC50 compared to a reference antibody, wherein the reference antibody is an antibody comprising a variable heavy chain indicated by 11C12 or SEQ ID NO: 1 and a variable light chain indicated by SEQ ID NO: 8, and optionally the reference antibody is of the same form, according to any one of claims 1 to 50.

52. An antibody or antigen-binding fragment thereof that binds to human EphA5 expressed on the surface of a cell, optionally on the surface of an H460 cell line, at an EC50 of less than or equal to 0.020 μg / mL, wherein optionally the EC50 is determined by flow cytometry, according to any one of claims 1 to 51.

53. An antibody or antigen-binding fragment thereof that binds to human EphA5 expressed on the surface of cells, optionally on the surface of H460 cell lines, at an EC50 of approximately 0.010 μg / mL to 0.020 μg / mL, wherein optionally the EC50 is determined by flow cytometry, according to any one of claims 1 to 52.

54. An antibody or antigen-binding fragment according to any one of claims 1 to 53, which binds to human EphA5 expressed on the surface of cells, optionally on the surface of H460 cell lines, at an EC50 of approximately 0.015 μg / mL, approximately 0.016 μg / mL, approximately 0.017 μg / mL, approximately 0.018 μg / mL, approximately 0.019 μg / mL, or approximately 0.020 μg / mL, wherein optionally the EC50 is determined by flow cytometry.

55. An antibody or antigen-binding fragment according to any one of claims 1 to 54, which is internalized by human EphA5-expressing cells.

56. A nucleic acid encoding the heavy chain of an antibody or an antigen-binding fragment according to any one of claims 1 to 55.

57. A nucleic acid encoding the light chain of an antibody or an antigen-binding fragment according to any one of claims 1 to 55.

58. A nucleic acid encoding the heavy chain and light chain of an antibody or antigen-binding fragment according to any one of claims 1 to 55.

59. A vector comprising the nucleic acid according to any one of claims 56 to 58.

60. The vector according to claim 59, which is an expression vector.

61. A vector comprising a nucleic acid encoding a heavy chain and a nucleic acid encoding a light chain, wherein the heavy chain and the light chain are antibodies or antigen-binding fragments according to any one of claims 1 to 58.

62. The vector according to claim 61, which is a bisistronic vector.

63. A vector system comprising a first vector containing a first nucleic acid encoding a heavy chain and a second vector containing a second nucleic acid encoding a light chain, wherein the heavy chain and the light chain are antibodies or antigen-binding fragments according to any one of claims 1 to 58.

64. The vector system according to claim 63, wherein the first vector and the second vector are expression vectors, respectively.

65. A host cell comprising the vector according to any one of claims 61 to 62 or the vector system according to any one of claims 63 to 64.

66. The host cell according to claim 65, which is a mammalian cell.

67. A method for producing an antibody, comprising the steps of introducing a vector according to any one of claims 61 to 62 and / or a vector system according to any one of claims 63 to 64 into a host cell, culturing the host cell under conditions for expressing an antibody or antigen-binding fragment from the host cell, and isolating or purifying the antibody or antigen-binding fragment.

68. The method according to claim 67, wherein the host cell is a mammalian cell.

69. An immunoconjugate having the formula Ab-(LD), a. Ab is an antibody or antigen-binding fragment according to any one of claims 1 to 55; b. L is a linker; and c. D is a cytotoxic drug. Immunoconjugate.

70. The immunoconjugate according to claim 69, wherein the linker is a severable linker.

71. The immunoconjugate according to claim 69 or claim 70, wherein the linker is a cathepsin-cleavable linker.

72. The immunoconjugate according to claim 71, wherein the cathepsin-cleavable linker comprises valine-citrulline (Val-Cit).

73. Linker, structure: An immunoconjugate according to any one of claims 68 to 72, which is an MC-VCP having

74. Linker, structure: An immunoconjugate according to any one of claims 69 to 72, including the immunoconjugate described in any one of claims 69 to 72.

75. The immunoconjugate according to any one of claims 68 to 74, wherein the linker is a pH-cleavable linker.

76. Linker, structure: The immunoconjugate according to any one of claims 69 to 79 and 75, which is a CL2A having

77. The immunoconjugate according to any one of claims 69 to 76, wherein the cytotoxic agent is auristatin.

78. Auristatin has the following structure: The immunoconjugate according to claim 77, which is monomethyl auristatin E (MMAE) having [a specific compound].

79. LD is structure: An immunoconjugate according to any one of claims 69 to 73, 77, and 78, comprising

80. LD is structure: An immunoconjugate according to any one of claims 69 to 72, 74, 77, and 78, comprising

81. An immunoconjugate having the formula Ab-(LD), a. Ab is an antibody that specifically binds to the human EPH receptor A5 (EphA5) epitope, comprising a heavy chain indicated by SEQ ID NO: 38 and a light chain indicated by SEQ ID NO: 40; and b. LD has the following structure: including, Immunoconjugate.

82. An immunoconjugate having the formula Ab-(LD), a. Ab is an antibody that specifically binds to the human EPH receptor A5 (EphA5) epitope, comprising a heavy chain indicated by SEQ ID NO: 39 and a light chain indicated by SEQ ID NO: 40; and b. LD has the following structure: including, Immunoconjugate.

83. Cytotoxic drugs, structure: An immunoconjugate according to any one of claims 69 to 76, wherein SN38 has the properties of SN38.

84. LD is structure: An immunoconjugate according to any one of claims 69, 70, 75, 76, and 83, comprising

85. LD is structure: An immunoconjugate according to any one of claims 69 to 72, 74, and 83, including the immunoconjugate described in any one of claims 69 to 72, 74, and 83.

86. An immunoconjugate according to any one of claims 69 to 85, wherein the drug-to-antibody ratio is approximately 4.

87. An immunoconjugate according to any one of claims 69 to 85, wherein the drug-to-antibody ratio is approximately 8.

88. A pharmaceutical composition comprising an antibody or antigen-binding fragment according to any one of claims 1 to 55 and a pharmaceutically acceptable carrier.

89. A pharmaceutical composition comprising an immunoconjugate according to any one of claims 69 to 87 and a pharmaceutically acceptable carrier.

90. An antibody or antigen-binding fragment thereof for use in the treatment, improvement, and / or prevention of cancer in a subject in need thereof, wherein the cancer is associated with the expression of EPH receptor A5, EPH receptor A5 is expressed on cancer cells, and the cancer is selected from the group consisting of lung cancer, breast cancer, esophageal cancer, gastric cancer, and ovarian cancer, according to any one of claims 1 to 55.

91. An immunoconjugate for use in the treatment, improvement, and / or prevention of cancer in a subject in need thereof, wherein the cancer is associated with the expression of EPH receptor A5, EPH receptor A5 is expressed on cancer cells, and the cancer is selected from the group consisting of lung cancer, breast cancer, esophageal cancer, gastric cancer, and ovarian cancer, according to any one of claims 69 to 87.

92. A pharmaceutical composition for use in the treatment, improvement, and / or prevention of cancer in a subject in need thereof, wherein the cancer is associated with the expression of EPH receptor A5, EPH receptor A5 is expressed on cancer cells, and the cancer is selected from the group consisting of lung cancer, breast cancer, esophageal cancer, gastric cancer, and ovarian cancer, according to claim 88.

93. A pharmaceutical composition for use in the treatment, improvement, and / or prevention of cancer in a subject in need thereof, wherein the cancer is associated with the expression of EPH receptor A5, EPH receptor A5 is expressed on cancer cells, and the cancer is selected from the group consisting of lung cancer, breast cancer, esophageal cancer, gastric cancer, and ovarian cancer, according to claim 89.

94. An antibody or antigen-binding fragment thereof for use in inducing tumor regression in a subject, wherein the tumor is cancer, the tumor is associated with the expression of EPH receptor A5, EPH receptor A5 is expressed on cancer cells, and the cancer is selected from the group consisting of lung cancer, breast cancer, esophageal cancer, gastric cancer, and ovarian cancer, according to any one of claims 1 to 55.

95. An immunoconjugate for use in inducing tumor regression in a subject requiring such induction, wherein the tumor is cancer, the tumor is associated with the expression of EPH receptor A5, EPH receptor A5 is expressed on cancer cells, and the cancer is selected from the group consisting of lung cancer, breast cancer, esophageal cancer, gastric cancer, and ovarian cancer, according to any one of claims 69 to 87.

96. A pharmaceutical composition for use in inducing tumor regression in a subject requiring such treatment, wherein the tumor is cancer, the tumor is associated with the expression of EPH receptor A5, EPH receptor A5 is expressed on cancer cells, and the cancer is selected from the group consisting of lung cancer, breast cancer, esophageal cancer, gastric cancer, and ovarian cancer.

97. A pharmaceutical composition for use in inducing tumor regression in a subject requiring such treatment, wherein the tumor is cancer, the tumor is associated with the expression of EPH receptor A5, EPH receptor A5 is expressed on cancer cells, and the cancer is selected from the group consisting of lung cancer, breast cancer, esophageal cancer, gastric cancer, and ovarian cancer.

98. A method for treating, improving, and / or preventing cancer in a subject in need thereof, comprising the step of administering a therapeutically effective amount of an antibody or antigen-binding fragment according to any one of claims 1 to 55 to the subject, wherein the cancer is associated with the expression of EPH receptor A5, EPH receptor A5 is expressed on cancer cells, and the cancer is selected from the group consisting of lung cancer, breast cancer, esophageal cancer, gastric cancer, and ovarian cancer.

99. A method for treating, improving, and / or preventing cancer in a subject in need thereof, comprising the step of administering a therapeutically effective dose of the immunoconjugate described in any one of claims 69 to 87 to the subject, wherein the cancer is associated with the expression of EPH receptor A5, EPH receptor A5 is expressed on cancer cells, and the cancer is selected from the group consisting of lung cancer, breast cancer, esophageal cancer, gastric cancer, and ovarian cancer.

100. A method for treating, improving, and / or preventing cancer in a subject in need, comprising the step of administering a therapeutically effective amount of the pharmaceutical composition according to claim 88 to the subject, wherein the cancer is associated with the expression of EPH receptor A5, EPH receptor A5 is expressed on cancer cells, and the cancer is selected from the group consisting of lung cancer, breast cancer, esophageal cancer, gastric cancer, and ovarian cancer.

101. A method for treating, improving, and / or preventing cancer in a subject in need, comprising the step of administering a therapeutically effective amount of the pharmaceutical composition according to claim 89 to the subject, wherein the cancer is associated with the expression of EPH receptor A5, EPH receptor A5 is expressed on cancer cells, and the cancer is selected from the group consisting of lung cancer, breast cancer, esophageal cancer, gastric cancer, and ovarian cancer.

102. A method for inducing tumor regression in a subject in need thereof, comprising the step of administering a therapeutically effective amount of an antibody or antigen-binding fragment according to any one of claims 1 to 55 to the subject, wherein the tumor is cancer, the tumor is associated with the expression of EPH receptor A5, EPH receptor A5 is expressed on cancer cells, and the cancer is selected from the group consisting of lung cancer, breast cancer, esophageal cancer, gastric cancer, and ovarian cancer.

103. A method for inducing tumor regression in a subject in need thereof, comprising the step of administering a therapeutically effective dose of the immunoconjugate described in any one of claims 69 to 87 to the subject, wherein the tumor is cancer, the tumor is associated with the expression of EPH receptor A5, EPH receptor A5 is expressed on cancer cells, and the cancer is selected from the group consisting of lung cancer, breast cancer, esophageal cancer, gastric cancer, and ovarian cancer.

104. A method for inducing tumor regression in a subject in need, comprising the step of administering a therapeutically effective amount of the pharmaceutical composition according to claim 88 to the subject, wherein the tumor is cancer, the tumor is associated with the expression of EPH receptor A5, EPH receptor A5 is expressed on cancer cells, and the cancer is selected from the group consisting of lung cancer, breast cancer, esophageal cancer, gastric cancer, and ovarian cancer.

105. A method for inducing tumor regression in a subject in need, comprising the step of administering a therapeutically effective amount of the pharmaceutical composition according to claim 89 to the subject, wherein the tumor is cancer, the tumor is associated with the expression of EPH receptor A5, EPH receptor A5 is expressed on cancer cells, and the cancer is selected from the group consisting of lung cancer, breast cancer, esophageal cancer, gastric cancer, and ovarian cancer.

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