ANTIPERIOSTIN ANTIBODIES AND THEIR USES

Abstract

This paper describes antibodies that block periostin function. It also describes their use in treating cancer and modifying the immune properties of tumors.

Claims

1. A recombinant antibody or antigen-binding fragment thereof that binds to periostin, characterized in that the antibody or antigen-binding fragment thereof comprises: a) an immunoglobulin heavy chain CDR1 (CDR-H1) comprising the amino acid sequence established at SEQ ID NO: 1 (GYTFTSYG); b) an immunoglobulin heavy chain CDR2 (CDR-H2) comprising an amino acid sequence established at any of SEQ ID NO: 2 (ISAYNGNT), 3 (ISAYSGNT), 4 (ISAYQGNT), 5 (ISAYTGNT) or 6 (ISAYDGNT); c) an immunoglobulin heavy chain CDR3 (CDR-H3) comprising an amino acid sequence established at any of SEQ ID NO: 7 (DILWPFDY), 8 (DVLWPFDY) or 9 (DMLWPFDY); d) an immunoglobulin light chain CDR1 (CDR-L1) comprising the amino acid sequence set out in SEQ ID NO: 10 (SSDIGSNR);e) an immunoglobulin light chain CDR2 (CDR-L2) comprising the amino acid sequence set out in SEQ ID NO: 11 (SND); and f) an immunoglobulin light chain CDR3 (CDR-L3) comprising the amino acid sequence set out in SEQ ID NO: 12 (AAWDDSLSTYV).; 2. A recombinant antibody or antigen-binding fragment thereof that binds to periostin, characterized in that the antibody or antigen-binding fragment thereof comprises any one, two, three, four, five, or six of: a) an immunoglobulin heavy chain CDR1 (CDR-H1) comprising the amino acid sequence set out in SEQ ID NO: 1 (GYTFTSYG); b) an immunoglobulin heavy chain CDR2 (CDR-H2) comprising an amino acid sequence set out in SEQ ID NO: 16 (ISAYXGNT); c) an immunoglobulin heavy chain CDR3 (CDR-H3) comprising an amino acid sequence set out in SEQ ID NO: 17 (DXLWPFDY); d) an immunoglobulin light chain CDR1 (CDR-L1) comprising the amino acid sequence set out in SEQ ID NO: 10 (SSDIGSNR); e) an immunoglobulin light chain CDR2 (CDR-L2) comprising the amino acid sequence set out in SEQ ID NO: 11 (SND);(f) an immunoglobulin light chain CDR3 (CDR-L3) comprising the amino acid sequence set out in SEQ ID NO: 12 (AAWDDSLSTYV); wherein X is any amino acid residue.; 3. The recombinant antibody or antigen-binding fragment thereof according to claim 1 or 2, further characterized in that the recombinant antibody or antigen-binding fragment thereof comprises: a) an immunoglobulin heavy chain CDR1 (CDR-H1) comprising the amino acid sequence stated in SEQ ID NO: 1 (GYTFTSYG); b) an immunoglobulin heavy chain CDR2 (CDR-H2) comprising the amino acid sequence eww nn / ίζηζ / E / γι stated in SEQ ID NO: 2 (ISAYNGNT); c) an immunoglobulin heavy chain CDR3 (CDR-H3) comprising the amino acid sequence stated in SEQ ID NO: 9 (DMLWPFDY); d) an immunoglobulin light chain CDR1 (CDR-L1) comprising the amino acid sequence stated in SEQ ID NO: 10 (SSDIGSNR); e) an immunoglobulin light chain CDR2 (CDR-L2) comprising the amino acid sequence set out in SEQ ID NO: 11 (SND);yf) an immunoglobulin light chain CDR3 (CDR-L3) comprising the amino acid sequence set out in SEQ ID NO: 12 (AAWDDSLSTYV).; 4. The recombinant antibody or antigen-binding fragment thereof according to any of claims 1 to 3, further characterized in that the recombinant antibody or antigen-binding fragment thereof is human, chimeric, or humanized.

5. The recombinant antibody or antigen-binding fragment thereof according to any of claims 1 to 4, further characterized in that the recombinant antibody or antigen-binding fragment thereof is an IgG antibody.

6. The recombinant antibody or antigen-binding fragment thereof according to any of claims 1 to 5, further characterized in that the recombinant antibody or antigen-binding fragment thereof comprises one or more mutations to reduce one or more effector functions of the recombinant antibody or antigen-binding fragment thereof.

7. The recombinant antibody or antigen-binding fragment thereof according to claim 6, further characterized in that the one or more mutations for reducing one or more effector functions of the recombinant antibody or antigen-binding fragment thereof comprise one or more mutations or sets of mutations selected from: N434A, N434H, T307A / E380A / N434A, M252Y / S254T / T256E, 433K / 434F / 436H, T250Q, T250F, M428L, M428F, T250Q / M428L, N434S, V308W, V308Y, V308F, M252Y / M428L, D259IA / 308F, M428LA / 308F, Q311V / N434S, T307Q / N434A, E258FA / 427T, S228P, L235E, S228P / L235E / R409K, S228P / L235E, K370Q, K370E, deletion of G446, deletion of K447, and combinations thereof of lgG4 according to the EU numbering system.

8. The recombinant antibody or antigen-binding fragment thereof according to claim 6, further characterized in that the one or more mutations to reduce one or more effector functions of the recombinant antibody or antigen-binding fragment thereof comprise: the S228P, F234A and L235A mutations of lgG4 according to the EU numbering system.

9. The recombinant antibody or antigen-binding fragment thereof according to any of claims 1 to 5, further characterized in that the recombinant antibody or antigen-binding fragment thereof is a Fab, F(ab)2, a single-domain antibody or a single-stranded variable fragment (scFv).

10. The recombinant antibody or antigen-binding fragment thereof according to any one of claims 1 to 9, further characterized in that it comprises an immunoglobulin heavy chain variable region and an immunoglobulin light chain variable region: a) wherein the immunoglobulin heavy chain variable region comprises an amino acid sequence that is at least approximately 90%, 95%, 97%, 99% or 100% identical to that set forth in SEQ ID NO: 13; and b) wherein the immunoglobulin light chain variable region comprises an amino acid sequence that is at least approximately 90%, 95%, 97%, 99% or 100% identical to that set forth in SEQ ID NO: 14; wherein amino acid residue number 55 of SEQ ID NO: 13 is asparagine, serine, glutamine, threonine or aspartic acid, or where amino acid residue number 100 of SEQ ID NO: 13 is methionine, isoleucine or valine.

11. The recombinant antibody or antigen-binding fragment thereof according to any of claims 1 to 10, further characterized in that the recombinant antibody or antigen-binding fragment thereof, when bound to periostin, comes into contact with one of the following amino acid residues: N276, R284, E288, L287, V295 and K302 of SEQ ID NO:

15.

12. The recombinant antibody or antigen-binding fragment thereof according to any of claims 1 to 11, further characterized in that the recombinant antibody or antigen-binding fragment thereof, when bound to periostin, comes into contact with two, three, four, or five of the following amino acid residues: N276, R284, E288, L287, V295, and K302 of SEQ ID NO:

15.

13. The recombinant antibody or antigen-binding fragment thereof according to any of claims 1 to 12, further characterized in that the recombinant antibody or antigen-binding fragment thereof, when bound to periostin, comes into contact with all of the following amino acid residues: N276, R284, E288, L287, V295 and K302 of SEQ ID NO:

15.

14. A recombinant antibody or antigen-binding fragment thereof that binds to periostin, characterized in that it comprises an immunoglobulin heavy chain variable region and an immunoglobulin light chain variable region: a) wherein the immunoglobulin heavy chain variable region comprises an amino acid sequence that is at least approximately 90%, 95%, 97%, 99% or 100% identical to that set forth in SEQ ID NO: 13; and b) wherein the immunoglobulin light chain variable region comprises an amino acid sequence that is at least approximately 90%, 95%, 97%, 99% or 100% identical to that set forth in SEQ ID NO: 14; wherein amino acid residue number 55 of SEQ ID NO: 13 is asparagine, serine, glutamine, threonine or aspartic acid, or where amino acid number 100 of SEQ ID NO: 13 is methionine, isoleucine or valine.

15. A recombinant antibody or antigen-binding fragment thereof that binds to periostin, characterized in that it comprises an immunoglobulin heavy chain variable region and an immunoglobulin light chain variable region: a) wherein the immunoglobulin heavy chain variable region comprises an amino acid sequence that is at least approximately 90%, 95%, 97%, 99% or 100% identical to that set out in SEQ ID NO: 13; and b) wherein the immunoglobulin light chain variable region comprises an amino acid sequence that is at least approximately 90%, 95%, 97%, 99% or 100% identical to that set out in SEQ ID NO:

14.

16. A recombinant antibody or antigen-binding fragment thereof that binds to periostin, characterized in that it comprises an immunoglobulin heavy chain variable region and an immunoglobulin light chain variable region: a) wherein the immunoglobulin heavy chain variable region comprises an amino acid sequence identical to that set out in SEQ ID NO: 13; and b) wherein the immunoglobulin light chain variable region comprises an amino acid sequence identical to that set out in SEQ ID NO:

14.

17. The recombinant antibody or antigen-binding fragment thereof according to any of claims 14 to 16, further characterized in that the recombinant antibody or antigen-binding fragment thereof is human, humanized, or chimeric.

18. The recombinant antibody or antigen-binding fragment thereof according to any of claims 14 to 17, further characterized in that the recombinant antibody or antigen-binding fragment thereof is an IgG antibody.

19. The recombinant antibody or antigen-binding fragment thereof according to any of claims 14 to 18, further characterized in that the recombinant antibody or antigen-binding fragment thereof comprises one or more mutations to reduce one or more effector functions of the recombinant antibody or antigen-binding fragment thereof.

20. The recombinant antibody or antigen-binding fragment thereof according to claim 19, further characterized in that the one or more mutations for reducing one or more effector functions of the recombinant antibody or antigen-binding fragment thereof comprise one or more mutations or sets of mutations selected from: N434A, N434H, T307A / E380A / N434A, M252Y / S254T / T256E, 433K / 434F / 436H, T250Q, T250F, M428L, M428F, T250Q / M428L, N434S, V308W, V308Y, V308F, M252Y / M428L, D259I / V308F, M428LA / 308F, Q311V / N434S, T307Q / N434A, E258FA / 427T, S228P, L235E, S228P / L235E / R409K, S228P / L235E, K370Q, K370E, deletion of G446, deletion of K447, and combinations of these of lgG4 according to the EU numbering system.

21. The recombinant antibody or antigen-binding fragment thereof according to claim 19, further characterized in that the mutation(s) to reduce one or more effector functions of the recombinant antibody or antigen-binding fragment thereof comprise: the S228P, F234A and L235A mutations of lgG4 according to the EU numbering system.

22. The recombinant antibody or antigen-binding fragment thereof according to any of claims 14 to 16, further characterized in that the recombinant antibody or antigen-binding fragment thereof is a Fab, F(ab)2, a single-domain antibody or a single-stranded variable fragment (scFv).

23. A recombinant antibody or antigen-binding fragment thereof characterized in that it binds to the fasciclin 2 (FAS2) domain of periostin.

24. The recombinant antibody or antigen-binding fragment thereof according to claim 23, further characterized in that the recombinant antibody or antigen-binding fragment thereof, when bound to periostin, comes into contact with an amino acid residue selected from amino acid 276 to 302 of SEQ ID NO:

15.

25. The recombinant antibody or antigen-binding fragment thereof according to claim 23 or 24, further characterized in that the recombinant antibody or antigen-binding fragment thereof, when bound to periostin, comes into contact with one of the following amino acid residues: N276, R284, E288, L287, V295 or K302 and SEQ ID NO:

15.

26. The recombinant antibody or antigen-binding fragment thereof according to any of claims 23 to 25, further characterized in that the recombinant antibody or antigen-binding fragment thereof, when bound to periostin, comes into contact with two, three, four, or five of the following amino acid residues: N276, R284, E288, L287, V295, and K302 of SEQ ID NO:

15.

27. The recombinant antibody or antigen-binding fragment thereof according to any of claims 23 to 26, further characterized in that the recombinant antibody or antigen-binding fragment thereof, when bound to periostin, comes into contact with all of the following amino acid residues: N276, R284, E288, L287, V295 or K302 and SEQ ID NO:

15.

28. The recombinant antibody or antigen-binding fragment thereof according to any of claims 23 to 27, further characterized in that the antibody or antigen-binding fragment thereof comprises: a) an immunoglobulin heavy chain CDR1 (CDR-H1) comprising the amino acid sequence established at SEQ ID NO: 1 (GYTFTSYG); b) an immunoglobulin heavy chain CDR2 (CDR-H2) comprising an amino acid sequence established at any of SEQ ID NO: 2 (ISAYNGNT), 3 (ISAYSGNT), 4 (ISAYQGNT), 5 (ISAYTGNT) or 6 (ISAYDGNT); c) an immunoglobulin heavy chain CDR3 (CDR-H3) comprising an amino acid sequence established at any of SEQ ID NO: 7 (DILWPFDY), 8 (DVLWPFDY) or 9 (DMLWPFDY); d) an immunoglobulin light chain CDR1 (CDR-L1) comprising the amino acid sequence set out in SEQ ID NO: 10 (SSDIGSNR);e) an immunoglobulin light chain CDR2 (CDR-L2) comprising the amino acid sequence set out in SEQ ID NO: 11 (SND); and f) an immunoglobulin light chain CDR3 (CDR-L3) comprising the amino acid sequence set out in SEQ ID NO: 12 (AAWDDSLSTYV).; 29. The recombinant antibody or antigen-binding fragment thereof according to any of claims 23 to 28, further characterized in that the recombinant antibody or antigen-binding fragment thereof is chimeric, humanized, or human.

30. The recombinant antibody or antigen-binding fragment thereof according to any of claims 23 to 29, further characterized in that the recombinant antibody or antigen-binding fragment thereof is an IgG antibody.

31. The recombinant antibody or antigen-binding fragment thereof according to any of claims 23 to 30, further characterized in that the recombinant antibody or antigen-binding fragment thereof comprises one or more mutations to reduce one or more effector functions of the recombinant antibody or antigen-binding fragment thereof.

32. The recombinant antibody or antigen-binding fragment thereof according to claim 31, further characterized in that the one or more mutations for reducing one or more effector functions of the recombinant antibody or antigen-binding fragment thereof comprise one or more mutations or sets of mutations selected from: N434A, N434H, T307A / E380A / N434A, M252Y / S254T / T256E, 433K / 434F / 436H, T250Q, T250F, M428L, M428F, T250Q / M428L, N434S, V308W, V308Y, V308F, M252Y / M428L, D259IA / 308F, M428LA / 308F, Q311V / N434S, T307Q / N434A, E258FA / 427T, S228P, L235E, S228P / L235E / R409K, S228P / L235E, K370Q, K370E, deletion of G446, deletion of K447, and combinations of these of lgG4 according to the EU numbering system.

33. The recombinant antibody or antigen-binding fragment thereof according to claim 31, further characterized in that the one or more mutations to reduce one or more effector functions of the recombinant antibody or antigen-binding fragment thereof comprise the S228P, F234A and L235A mutations of lgG4 according to the EU numbering system.

34. The recombinant antibody or antigen-binding fragment thereof according to any of claims 23 to 30, further characterized in that the recombinant antibody or antigen-binding fragment thereof is a Fab, F(ab)2, a single-domain antibody or a single-stranded variable fragment (scFv).

35. The recombinant antibody or antigen-binding fragment thereof according to any of claims 1 to 34, further characterized in that the antibody has an IC50 of less than approximately 50 nanomolar in a cell adhesion assay performed with human lung fibroblast cells and / or mouse fibroblast cells.

36. A nucleic acid characterized in that it encodes the recombinant antibody or antigen-binding fragment thereof as claimed in any of claims 1 to 35.

37. A cell line characterized in that it comprises the nucleic acid as claimed in claim 36.

38. The cell line according to claim 37, further characterized in that the cell line is a Chinese hamster ovary cell line.

39. A pharmaceutical composition characterized in that it comprises the recombinant antibody or antigen-binding fragment thereof as claimed in any of claims 1 to 35 and a pharmaceutically acceptable excipient, carrier or diluent.

40. The pharmaceutical composition according to claim 39, characterized in that it is formulated to be administered intravenously.

41. The pharmaceutical composition according to claim 39, characterized in that it is formulated to be administered subcutaneously.

42. The pharmaceutical composition according to claim 39, characterized in that it is formulated for intratumoral administration. eww nn / Lznz / E / Yi 43. Use of a recombinant antibody or antigen-binding fragment thereof as claimed in any of claims 1 to 35 or a pharmaceutical composition as claimed in any of claims 39 to 42 for the manufacture of a medicament for reducing the collagen content in a tumor in an individual, wherein the recombinant antibody or antigen-binding fragment thereof or the pharmaceutical composition is adapted to be administered to said individual.

44. The recombinant antibody or antigen-binding fragment thereof as claimed in any of claims 1 to 35 or the pharmaceutical composition as claimed in any of claims 39 to 42 for use in reducing collagen content in a tumor.

45. Use of a recombinant antibody or antigen-binding fragment thereof as claimed in any of claims 1 to 35 or a pharmaceutical composition as claimed in any of claims 39 to 42 for the manufacture of a medicament for treating cancer in an individual, wherein the recombinant antibody or antigen-binding fragment thereof or the pharmaceutical composition is adapted to be administered to such individual.

46. ​​Use according to claim 45, wherein cancer comprises glioblastoma, pancreatic cancer, breast cancer, bladder cancer, kidney cancer, head and neck cancer, ovarian cancer, skin cancer, stomach cancer, mesothelioma, liver cancer, endometrial cancer, colon cancer, cervical cancer, prostate cancer, or lung cancer.

47. The recombinant antibody or antigen-binding fragment thereof as claimed in any of claims 1 to 35 or the pharmaceutical composition as claimed in any of claims 39 to 42 for use in the treatment of cancer.

48. Use according to claim 47, wherein cancer comprises glioblastoma, pancreatic cancer, breast cancer, bladder cancer, kidney cancer, head and neck cancer, ovarian cancer, skin cancer, stomach cancer, mesothelioma, liver cancer, endometrial cancer, colon cancer, cervical cancer, prostate cancer, or lung cancer.

49. A recombinant antibody or antigen-binding fragment thereof as claimed in any of claims 1 to 35 or a pharmaceutical composition as claimed in any of claims 39 to 42 for the manufacture of a medicament for increasing the M1 macrophage phenotype and / or reducing the M2 macrophage phenotype in a tumor in an individual, wherein the recombinant antibody or antigen-binding fragment thereof or the pharmaceutical composition is adapted to be administered to said individual.

50. The recombinant antibody or antigen-binding fragment thereof as claimed in any of claims 1 to 35 or the pharmaceutical composition as claimed in any of claims 39 to 42 for use in increasing the M1 macrophage phenotype and / or reducing the M2 macrophage phenotype in a tumor.

51. A recombinant antibody or antigen-binding fragment thereof as claimed in any of claims 1 to 35 or a pharmaceutical composition as claimed in any of claims 39 to 42 for the manufacture of a medicament for reducing the accumulation of myeloid granulocytic suppressor cells and / or tumor-associated macrophages in an individual, wherein the recombinant antibody or antigen-binding fragment thereof or the pharmaceutical composition is adapted to be administered to said individual.

52. The recombinant antibody or antigen-binding fragment thereof as claimed in any of claims 1 to 35 or the pharmaceutical composition as claimed in any of claims 39 to 42 for use in reducing the accumulation of myeloid granulocytic suppressor cells and / or tumor-associated macrophages in a tumor.

53. A recombinant antibody or antigen-binding fragment thereof as claimed in any of claims 1 to 35 or a pharmaceutical composition as claimed in any of claims 39 to 42 for the manufacture of a medicament for increasing the frequency of CD4+ and / or CD8+ T lymphocytes in a tumor of an individual, wherein the recombinant antibody or antigen-binding fragment thereof or the pharmaceutical composition is adapted to be administered to said individual.

54. The recombinant antibody or antigen-binding fragment thereof as claimed in any of claims 1 to 35 or the pharmaceutical composition as claimed in any of claims 39 to 42 for use in increasing the frequency of CD4+ and / or CD8+ T lymphocytes in a tumor.

55. A recombinant antibody or antigen-binding fragment thereof as claimed in any of claims 1 to 35 or a pharmaceutical composition as claimed in any of claims 39 to 42 for the manufacture of a medicament for increasing the frequency of CD8+ T lymphocytes in a tumor in an individual, wherein the recombinant antibody or antigen-binding fragment thereof or the pharmaceutical composition is adapted to be administered to said individual.

56. The recombinant antibody or antigen-binding fragment thereof as claimed in any of claims 1 to 35 or the pharmaceutical composition as claimed in any of claims 39 to 42 for use in increasing the frequency of CD8+ T lymphocytes in a tumor.

57. A recombinant antibody or antigen-binding fragment thereof as claimed in any of claims 1 to 35 or a pharmaceutical composition as claimed in any of claims 39 to 42 for the manufacture of a medicament for augmenting the function of CD8+ T lymphocytes as measured by the expression and / or release of interferon gamma by CD8+ T lymphocytes in an individual, wherein the recombinant antibody or antigen-binding fragment thereof or the pharmaceutical composition is adapted to be administered to such individual.

58. The recombinant antibody or antigen-binding fragment thereof as claimed in any of claims 1 to 35 or the pharmaceutical composition as claimed in any of claims 39 to 42 for use in enhancing the function of CD8+ T lymphocytes as measured by the expression and / or release of interferon gamma by CD8+ T lymphocytes.

59. A method for preparing a composition for decreasing the collagen content in a tumor, characterized in that it comprises mixing the recombinant antibody or antigen-binding fragment of the same as claimed in any of claims 1 to 35 and a pharmaceutically acceptable excipient, carrier, or diluent.

60. A method for preparing a composition for increasing the M1 macrophage phenotype and / or reducing the M2 macrophage phenotype in a tumor, characterized in that it comprises mixing the recombinant antibody or antigen-binding fragment thereof as claimed in any of claims 1 to 35 and a pharmaceutically acceptable excipient, carrier, or diluent.

61. A method for preparing a composition for reducing the accumulation of myeloid granulocytic suppressor cells and / or tumor-associated macrophages in an individual, characterized in that it comprises mixing the recombinant antibody or antigen-binding fragment thereof as claimed in any of claims 1 to 35 and a pharmaceutically acceptable excipient, carrier, or diluent.

62. A method for preparing a composition for increasing the frequency of CD4+ and / or CD8+ T lymphocytes in a tumor of an individual, characterized in that it comprises mixing the recombinant antibody or antigen-binding fragment thereof as claimed in any of claims 1 to 35 and a pharmaceutically acceptable excipient, carrier or diluent.

63. A method for preparing a composition for increasing the expression and / or release of interferon gamma by CD8+ T lymphocytes in a tumor in an individual, characterized in that it comprises mixing the recombinant antibody or antigen-binding fragment thereof as claimed in any of claims 1 to 35 and a pharmaceutically acceptable excipient, carrier or diluent.

64. A method for preparing a composition for treating cancer, characterized in that it comprises mixing the recombinant antibody or antigen-binding fragment thereof as claimed in any of claims 1 to 35 and a pharmaceutically acceptable excipient, carrier, or diluent.

65. The method according to claim 64, further characterized in that the cancer comprises glioblastoma, pancreatic cancer, breast cancer, bladder cancer, kidney cancer, head and neck cancer, ovarian cancer, skin cancer, stomach cancer, mesothelioma, liver cancer, endometrial cancer, colon cancer, cervical cancer, prostate cancer, or lung cancer.

66. A method for producing the recombinant antibody or antigen-binding fragment thereof as claimed in any one of claims 1 to 35, characterized in that it comprises incubating the cell line as claimed in claims 37 or 38 in a cell culture medium under conditions sufficient to permit the expression and secretion of the recombinant antibody or antigen-binding fragment thereof as claimed in any one of claims 1 to 35.