ANTI-HSPA5 (GRP78) ANTIBODIES AND USES THEREOF

• US 20190330318A1
• Filed: 07/24/2017
• Published: 10/31/2019
• Est. Priority Date: 07/25/2016
• Status: Abandoned Application

• Current Assignee: Biogen MA Inc. (Biogen, Inc.), The Board of Regents of the University of Colorado (University of Colorado System)
• Original Assignee: Biogen MA Inc. (Biogen, Inc.), The Board of Regents of the University of Colorado (University of Colorado System)
• Inventors: Bennett, Jeffrey L., Ransohoff, Richard M., Shimizu, Fumitaka, Schaller, Kristin L.
• Application Number:
  US16/319,715
• Publication Number:
  US 20190330318A1
• Time in Patent Office: Days
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 2039/507 Comprising a combination of two or more separate antibodies
• CPC Class Codes: A61K 2039/54 characterised by the route of administration
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: C07K 16/18 against material from animals or humans
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype
• CPC Class Codes: C07K 2317/56 variable (Fv) region, i.e. VH and/or VL
• CPC Class Codes: C07K 2317/75 Agonist effect on antigen
• CPC Class Codes: C12N 15/85 for animal cells
• CPC Class Codes: G01N 2800/24 Immunology or allergic disorders
• CPC Class Codes: G01N 2800/52 Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
• CPC Class Codes: G01N 33/6854 Immunoglobulins

Abstract

This disclosure provides anti-HSPA5 antibodies. Also featured is a method for delivering an agent into the brain, spinal cord, or other component of the central nervous system. In addition, methods of identifying a human subject at risk of relapse of neuromyelitis optica (NMO) or neuropsychiatric systemic lupus erythematosus (NP-SLE) are disclosed, as are methods of determining the severity of an attack of NMO or NP-SLE, and methods of diagnosing NP-SLE.

Claims

1. An isolated antibody or antigen-binding fragment thereof that specifically binds human heat shock protein family A (Hsp70) member 5 (human HSPA5), wherein the antibody or antigen-binding fragment thereof:

(i) binds to the same epitope on human HSPA5 as a reference antibody;

or (ii) competes with the reference antibody to bind to human HSPA5, wherein the reference antibody comprises;

(a) a heavy chain variable region (VH) comprising VH complementarity determining regions 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NO;

1, SEQ ID NO;

2, SEQ ID NO;

3, respectively, and a light chain variable region (VL) comprising VL complementarity determining regions 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NO;

4, SEQ ID NO;

5, and SEQ ID NO;

6, respectively;

or

(b) a VH comprising VH complementarity determining regions 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NO;

11, SEQ ID NO;

12, SEQ ID NO;

13, respectively, and a VL comprising VL complementarity determining regions 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NO;

14, SEQ ID NO;

15, and SEQ ID NO;

16, respectively.

2. The antibody or antigen-binding fragment of claim 1, wherein:

(a) the VH of the reference antibody consists of SEQ ID NO;

7 and the VL of the reference antibody consists of SEQ ID NO;

8;

or

(b) the VH of the reference antibody consists of SEQ ID NO;

17 and the VL of the reference antibody consists of SEQ ID NO;

18.

3. The antibody or antigen-binding fragment of claim 1, wherein the reference antibody comprises a heavy chain and a light chain, wherein:

(a) the heavy chain consists of SEQ ID NO;

9 and the light chain consists of SEQ ID NO;

10;

or

(b) the heavy chain consists of SEQ ID NO;

19 and the light chain consists of SEQ ID NO;

20.

4. The antibody or antigen-binding fragment of any one of claims 1 to 3, wherein the antibody comprises a heavy chain of a human IgG isotype.

5. The antibody or antigen-binding fragment of claim 4, wherein the human IgG isotype is selected from the group consisting of IgG1, IgG2a, IgG2b, IgG3, and IgG4.

6. The antibody or antigen-binding fragment of any one of claims 1 to 5, wherein the antibody comprises a light chain of a human lambda isotype.

7. The antibody or antigen-binding fragment of any one of claims 1 to 5, wherein the antibody comprises a light chain of a human kappa isotype.

8. The antibody or antigen-binding fragment of any one of claims 1 to 3, wherein the antigen-binding fragment is selected from the group consisting of an scFv, an sc(Fv)2, an Fab, an F(ab)2, a diabody, a humanized nanobody, and a humanized Variable domain of New Antigen Receptor (VNAR).

9. The antibody or antigen-binding fragment of any one of claims 1 to 8, wherein the antibody or antigen-binding fragment is human or humanized.

51. A pharmaceutical composition comprising the antibody or antigen-binding fragment of any one of claims 1 to 22.

55. The method of any one of claims 52 to 54, wherein the antibody or antigen-binding fragment is an antibody or antigen-binding fragment of any one of claims 1 to 22.

10. An isolated antibody or antigen-binding fragment thereof that specifically binds human HSPA5, wherein the antibody or antigen-binding fragment thereof comprises:

(a) a heavy chain variable region (VH) comprising VH complementarity determining regions 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NO;

1, SEQ ID NO;

2, SEQ ID NO;

3, respectively, and a light chain variable region (VL) comprising VL complementarity determining regions 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NO;

4, SEQ ID NO;

5, and SEQ ID NO;

6, respectively;

or

(b) a VH comprising VH complementarity determining regions 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NO;

11, SEQ ID NO;

12, SEQ ID NO;

13, respectively, and a VL comprising VL complementarity determining regions 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NO;

14, SEQ ID NO;

15, and SEQ ID NO;

16, respectively.

11. The antibody or antigen-binding fragment of claim 10, wherein:

(a) the VH consists of an amino acid sequence that is at least 75% identical to the amino acid sequence set forth in SEQ ID NO;

7 and the VL consists of an amino acid sequence that is at least 75% identical to the amino acid sequence set forth in SEQ ID NO;

8;

or

(b) the VH consists of an amino acid sequence that is at least 75% identical to the amino acid sequence set forth in SEQ ID NO;

17 and the VL consists of an amino acid sequence that is at least 75% identical to the amino acid sequence set forth in SEQ ID NO;

18.

12. The antibody or antigen-binding fragment of claim 10, wherein:

(a) the VH consists of an amino acid sequence that is at least 95% identical to the amino acid sequence set forth in SEQ ID NO;

7 and the VL consists of an amino acid sequence that is at least 95% identical to the amino acid sequence set forth in SEQ ID NO;

8, wherein the VH and VL are not identical to the amino acid sequence set forth in SEQ ID NOs;

7 and 8, respectively;

or

(b) the VH consists of an amino acid sequence that is at least 95% identical to the amino acid sequence set forth in SEQ ID NO;

17 and the VL consists of an amino acid sequence that is at least 95% identical to the amino acid sequence set forth in SEQ ID NO;

18, wherein the VH and VL are not identical to the amino acid sequence set forth in SEQ ID NOs;

17 and 18, respectively.

13. The antibody or antigen-binding fragment of claim 10, wherein:

(a) the VH consists of SEQ ID NO;

7 and the VL consists of SEQ ID NO;

8;

or

(b) the VH consists of SEQ ID NO;

17 and the VL consists of SEQ ID NO;

18.

14. The antibody or antigen-binding fragment of claim 10, wherein the antibody or antigen-binding fragment comprises a heavy chain and a light chain, wherein:

(a) the heavy chain consists of an amino acid sequence that is at least 75% identical to the amino acid sequence set forth in SEQ ID NO;

9 and the light chain consists of an amino acid sequence that is at least 75% identical to the amino acid sequence set forth in SEQ ID NO;

10;

or

(b) the heavy chain consists of an amino acid sequence that is at least 75% identical to the amino acid sequence set forth in SEQ ID NO;

19 and the light chain consists of an amino acid sequence that is at least 75% identical to the amino acid sequence set forth in SEQ ID NO;

20.

15. The antibody or antigen-binding fragment of claim 10, wherein the antibody comprises a heavy chain and a light chain, wherein:

(a) the heavy chain consists of an amino acid sequence that is at least 95% identical to the amino acid sequence set forth in SEQ ID NO;

9 and the light chain consists of an amino acid sequence that is at least 95% identical to the amino acid sequence set forth in SEQ ID NO;

10, wherein the heavy chain and light chain are not identical to the amino acid sequence set forth in SEQ ID NOs;

9 and 10, respectively;

or

(b) the heavy chain consists of an amino acid sequence that is at least 95% identical to the amino acid sequence set forth in SEQ ID NO;

19 and the light chain consists of an amino acid sequence that is at least 95% identical to the amino acid sequence set forth in SEQ ID NO;

20, wherein the heavy chain and light chain are not identical to the amino acid sequence set forth in SEQ ID NOs;

19 and 20, respectively.

16. The antibody or antigen-binding fragment of claim 10, wherein the antibody comprises a heavy chain and a light chain, wherein:

(a) the heavy chain consists of SEQ ID NO;

9 and the light chain consists of SEQ ID NO;

10;

or

(b) the heavy chain consists of SEQ ID NO;

19 and the light chain consists of SEQ ID NO;

20.

17. The antibody or antigen-binding fragment of any one of claims 10 to 13, wherein the antibody comprises a heavy chain of a human IgG isotype.

18. The antibody or antigen-binding fragment of claim 17, wherein the human IgG isotype is selected from the group consisting of IgG1, IgG2a, IgG2b, IgG3, and IgG4.

19. The antibody or antigen-binding fragment of any one of claims 10 to 13, wherein the antibody comprises a light chain of a human lambda isotype.

20. The antibody or antigen-binding fragment of any one of claims 10 to 13, wherein the antibody comprises a light chain of a human kappa isotype.

21. The antibody or antigen-binding fragment of any one of claims 10 to 13, wherein the antigen-binding fragment is selected from the group consisting of an scFv, an sc(Fv)2, an Fab, an F(ab)2, a diabody, a humanized nanobody, and a humanized Variable domain of New Antigen Receptor (VNAR).

22. The antibody or antigen-binding fragment of any one of claims 10 to 21, wherein the antibody or antigen-binding fragment is human or humanized.

23. An expression vector comprising a promoter operably linked to a polynucleotide encoding a polypeptide comprising:

(i) an immunoglobulin heavy chain variable region (VH) comprising VH complementarity determining regions (CDRs) 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NOs;

1-3, respectively, wherein the VH when paired with a light chain variable region (VL) comprising the amino acid sequence set forth in SEQ ID NO;

8 binds human HSPA5;

(ii) an immunoglobulin VL comprising VL CDRs 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NOs;

4-6, respectively, wherein the VL when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO;

7 binds to human HSPA5;

(iii) an immunoglobulin VH comprising VH CDRs 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NOs;

11-13, respectively, wherein the VH when paired with a VL comprising the amino acid sequence set forth in SEQ ID NO;

18 binds human HSPA5;

or

(iv) an immunoglobulin VL comprising VL CDRs 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NOs;

14-16, respectively, wherein the VL when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO;

17 binds to human HSPA5.

24. The expression vector of claim 23, wherein the promoter is a heterologous promoter.

25. The expression vector of claim 24, wherein the heterologous promoter is a cytomegalovirus, simian virus 40, or retroviral promoter.

26. The expression vector of claim 24, wherein the heterologous promoter is the cytomegalovirus immediate early promoter.

27. The expression vector of claim 23, wherein the expression vector comprises cytomegalovirus intron-A.

28. The expression vector of claim 23, wherein the polypeptide comprises a signal peptide.

29. The expression vector of claim 28, wherein the signal peptide is a heterologous signal peptide.

30. The expression vector of any one of claims 23 to 29, wherein the polypeptide comprises:

(i) an immunoglobulin heavy chain or fragment thereof comprising a VH with the amino acid sequence set forth in SEQ ID NO;

7;

(ii) an immunoglobulin light chain or fragment thereof comprising a VL with the amino acid sequence set forth in SEQ ID NO;

8;

(iii) an immunoglobulin heavy chain or fragment thereof comprising a VH with the amino acid sequence set forth in SEQ ID NO;

17;

or

(iv) an immunoglobulin light chain or fragment thereof comprising a VL with the amino acid sequence set forth in SEQ ID NO;

18.

31. The expression vector of any one of claims 23 to 29, wherein the polynucleotide comprises the nucleic acid sequence set forth in:

SEQ ID NO;

21;

SEQ ID NO;

22;

SEQ ID NO;

23;

or

SEQ ID NO;

24.

32. The expression vector of any one of claims 23 to 31, wherein the expression vector is a plasmid, phage, virus, or retrovirus.

33. An expression vector comprising:

(a) a first polynucleotide encoding a first polypeptide comprising an immunoglobulin heavy chain or a fragment thereof comprising a heavy chain variable region (VH) comprising VH complementarity determining regions (CDRs) 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NOs;

1-3, respectively; and

a second polynucleotide encoding a second polypeptide comprising an immunoglobulin light chain or a fragment thereof comprising a light chain variable region (VL) comprising VL CDRs 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NOs;

4-6, respectively, wherein the immunoglobulin heavy chain or fragment thereof when paired with the immunoglobulin light chain or fragment thereof forms an anti-human HSPA5 antibody or human HSPA5-binding fragment thereof, or

(b) a first polynucleotide encoding a first polypeptide comprising an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising VH CDRs 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NOs;

11-13, respectively; and

a second polynucleotide encoding a second polypeptide comprising an immunoglobulin light chain or a fragment thereof comprising a VL comprising VL CDRs 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NOs;

14-16, respectively, wherein the immunoglobulin heavy chain or fragment thereof when paired with the immunoglobulin light chain or fragment thereof forms an anti-human HSPA5 antibody or human HSPA5-binding fragment thereof.

34. The expression vector of claim 33, wherein the immunoglobulin heavy chain comprises a human IgG1 heavy chain constant region and the immunoglobulin light chain comprises a human lambda light chain constant region.

35. The expression vector of claim 33, wherein the immunoglobulin heavy chain comprises a human IgG1 heavy chain constant region and the immunoglobulin light chain comprises a human kappa light chain constant region.

36. The expression vector of any one of claims 33 to 35, wherein:

(a) the VH consists of the amino acid sequence set forth in SEQ ID NO;

7, and the VL consists of the amino acid sequence set forth in SEQ ID NO;

8;

or

(b) the VH consists of the amino acid sequence set forth in SEQ ID NO;

17, and the VL consists of the amino acid sequence set forth in SEQ ID NO;

18.

37. The expression vector of claim 33, wherein:

(a) the heavy chain consists of SEQ ID NO;

9 and the light chain consists of SEQ ID NO;

10;

or

(b) the heavy chain consists of SEQ ID NO;

19 and the light chain consists of SEQ ID NO;

20.

38. The expression vector of claim 33, wherein:

(a) the first polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO;

21, and the second polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO;

22;

or

(b) the first polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO;

23, and the second polynucleotide comprises the nucleic acid sequence set forth in SEQ ID NO;

24.

39. The expression vector of any one of claims 33 to 38, wherein the expression vector is a plasmid, phage, virus, or retrovirus.

40. A cDNA comprising a polynucleotide encoding a polypeptide comprising:

(i) an immunoglobulin heavy chain variable region (VH) comprising VH complementarity determining regions (CDRs) 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NOs;

1-3, respectively, wherein the VH when paired with a light chain variable region (VL) comprising the amino acid sequence set forth in SEQ ID NO;

8 binds human HSPA5;

(ii) an immunoglobulin VL comprising VL CDRs 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NOs;

4-6, respectively, wherein the VL when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO;

7 binds human HSPA5;

(iii) an immunoglobulin VH comprising VH complementarity determining regions (CDRs) 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NOs;

11-13, respectively, wherein the VH when paired with a VL comprising the amino acid sequence set forth in SEQ ID NO;

18 binds human HSPA5;

or

(iv) an immunoglobulin VL comprising VL CDRs 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NOs;

14-16, respectively, wherein the VL when paired with a VH comprising the amino acid sequence set forth in SEQ ID NO;

17 binds human HSPA5.

41. A host cell comprising:

(a) a first expression vector comprising a first polynucleotide encoding a first polypeptide comprising an immunoglobulin heavy chain or a fragment thereof comprising a heavy chain variable region (VH) comprising VH complementarity determining regions (CDRs) 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NOs;

1-3, respectively, and a second expression vector comprising a second polynucleotide encoding a second polypeptide comprising an immunoglobulin light chain or a fragment thereof comprising a light chain variable region (VL) comprising VL CDRs 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NOs;

4-6, respectively, wherein the immunoglobulin heavy chain or fragment thereof when paired with the immunoglobulin light chain or fragment thereof forms an anti-human HSPA5 antibody or human HSPA5-binding fragment thereof;

or

(b) a first expression vector comprising a first polynucleotide encoding a first polypeptide comprising an immunoglobulin heavy chain or a fragment thereof comprising a VH comprising VH CDRs 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NOs;

11-13, respectively, and a second expression vector comprising a second polynucleotide encoding a second polypeptide comprising an immunoglobulin light chain or a fragment thereof comprising a VL comprising VL CDRs 1, 2, and 3 with the amino acid sequences set forth in SEQ ID NOs;

14-16, respectively, wherein the immunoglobulin heavy chain or fragment thereof when paired with the immunoglobulin light chain or fragment thereof forms an anti-human HSPA5 antibody or human HSPA5-binding fragment thereof.

42. The host cell of claim 41, wherein the immunoglobulin heavy chain comprises a human IgG1 heavy chain constant region and the immunoglobulin light chain comprises a human lambda light chain constant region.

43. The host cell of claim 41, wherein the immunoglobulin heavy chain comprises a human IgG1 heavy chain constant region and the immunoglobulin light chain comprises a human kappa light chain constant region.

44. The host cell of any one of claims 41 to 43, wherein:

(a) the VH consists of the amino acid sequence set forth in SEQ ID NO;

7, and the VL consists of the amino acid sequence set forth in SEQ ID NO;

8;

or

(b) the VH consists of the amino acid sequence set forth in SEQ ID NO;

17, and the VL consists of the amino acid sequence set forth in SEQ ID NO;

18.

45. The host cell of claim 41, wherein:

(a) the heavy chain consists of SEQ ID NO;

9 and the light chain consists of SEQ ID NO;

10;

or

(b) the heavy chain consists of SEQ ID NO;

19 and the light chain consists of SEQ ID NO;

20.

46. The host cell of any one of claims 41 to 45, wherein the host cell is a mammalian host cell.

47. The host cell of any one of claims 41 to 45, wherein the host cell is a Chinese Hamster Ovary (CHO) cell, a HEK 293 cell, or a NSO cell.

48. A method of making an antibody or antigen-binding fragment thereof that specifically binds human HSPA5, the method comprising:

culturing the host cell of any one of claims 41 to 47 in a cell culture; and

isolating the antibody or antigen-binding fragment from the cell culture.

49. The method of claim 48, further comprising formulating the antibody or antigen-binding fragment with a pharmaceutically acceptable carrier into a sterile pharmaceutical composition suitable for administration to a human subject.

50. The method of claim 49, wherein the pharmaceutical composition is suitable for intravenous or subcutaneous administration.

52. A method of delivering an agent to the brain, spinal cord, or other component of the central nervous system of a human subject in need thereof, the method comprising administering to the subject the agent and an antibody or antigen-binding fragment thereof that specifically binds human HSPA5.

53. The method of claim 52, wherein the antibody or antigen-binding fragment and the agent are administered simultaneously.

54. The method of claim 52, wherein the antibody or antigen-binding fragment and the agent are administered sequentially.

56. The method of any one of claims 52 to 55, wherein the agent is selected from the group consisting of an antibody or antigen-binding fragment thereof, a small molecule, siRNA, a degron, and a protein mimotope.

57. The method of claim 56, wherein the agent is an antibody or antigen-binding fragment thereof selected from the group consisting of:

an antibody or antigen-binding fragment that specifically binds tau,

an antibody or antigen-binding fragment that specifically binds β

-amyloid,

an antibody or antigen-binding fragment that specifically binds α

-synuclein,

an antibody or antigen-binding fragment that specifically binds TDP-43,

an antibody or antigen-binding fragment that specifically binds AQP4,

an antibody or antigen-binding fragment that specifically binds IL6R,

an antibody or antigen-binding fragment that specifically binds CD20,

an antibody or antigen-binding fragment that specifically binds CD25,

an antibody or antigen-binding fragment that specifically binds VEGF-A,

an antibody or antigen-binding fragment that specifically binds BAFF,

an antibody or antigen-binding fragment that specifically binds alpha-4-integrin,

an antibody or antigen-binding fragment that specifically binds human complement protein C3,

an antibody or antigen-binding fragment that specifically binds human complement protein C1q, and

an antibody or antigen-binding fragment that specifically binds human complement protein C5.

58. The method of claim 56, wherein the agent is a small molecule that is a small molecule chemotherapeutic.

59. A method of identifying a human subject at risk of relapse of neuromyelitis optica (NMO), the method comprising:

measuring the titer of anti-HSPA5 antibodies in the human subject; and

determining that the titer of anti-HSPA5 antibodies in the human subject are higher than a control level.

60. A method of determining the severity of an attack of neuromyelitis optica (NMO) in a human subject, the method comprising:

measuring the titer of anti-HSPA5 antibodies in the human subject; and

comparing the titer of anti-HSPA5 antibodies in the human subject to a control level.

61. A method of diagnosing a human subject with neuropsychiatric systemic lupus erythematosus (NP-SLE), the method comprising:

measuring the titer of anti-HSPA5 antibodies in the human subject; and

determining that the titer of anti-HSPA5 antibodies in the human subject are higher than a control level.

62. A method of identifying a human subject at risk of relapse of neuropsychiatric systemic lupus erythematosus (NP-SLE), the method comprising:

measuring the titer of anti-HSPA5 antibodies in the human subject; and

determining that the titer of anti-HSPA5 antibodies in the human subject are higher than a control level.

63. A method of determining the severity of an attack of neuropsychiatric systemic lupus erythematosus (NP-SLE) in a human subject, the method comprising:

measuring the titer of anti-HSPA5 antibodies in the human subject; and

comparing the titer of anti-HSPA5 antibodies in the human subject to a control level.