CHIMERIC RECEPTORS AND USES THEREOF IN IMMUNE THERAPY

• US 20150139943A1
• Filed: 10/17/2014
• Published: 05/21/2015
• Est. Priority Date: 10/17/2013
• Status: Active Grant

• Current Assignee: St Jude Children's Research Hospital Incorporated (Abbott Laboratories Incorporated), Cogent Biosciences, Inc., National University of Singapore
• Original Assignee: St Jude Children's Research Hospital Incorporated (Abbott Laboratories Incorporated), National University of Singapore
• Inventors: Campana, Dario, Kudo, Ko
• Granted Patent: US 10,144,770 B2
• Time in Patent Office: Days
• US Class Current: 424/85.2
• CPC Class Codes: A61K 2039/515 Animal cells
• CPC Class Codes: A61K 2039/572 cytotoxic response
• CPC Class Codes: A61K 2239/31 characterized by the route of administration
• CPC Class Codes: A61K 2239/38 characterised by the dose, timing or administration schedule
• CPC Class Codes: A61K 2300/00 Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
• CPC Class Codes: A61K 35/17 Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes when activated by a specific antigen A61K39/00
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 39/39558 against tumor tissues, cells, antigens
• CPC Class Codes: A61K 39/4611 T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
• CPC Class Codes: A61K 39/4631 Chimeric Antigen Receptors [CAR]
• CPC Class Codes: A61K 39/464429 Molecules with a "CD" designation not provided for elsewhere
• CPC Class Codes: C07K 14/7051 T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 14/70517 CD8
• CPC Class Codes: C07K 14/70535 Fc-receptors, e.g. CD16, CD32, CD64 (CD2314/705F)
• CPC Class Codes: C07K 14/70578 NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95 NGF-receptor C07K14/71, TNF-receptor C07K14/7151
• CPC Class Codes: C07K 16/2887 against CD20
• CPC Class Codes: C07K 16/3084 against tumour-associated gangliosides
• CPC Class Codes: C07K 16/32 against translation products of oncogenes
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/02 containing a signal sequence
• CPC Class Codes: C07K 2319/03 : containing a transmembrane segment
• CPC Class Codes: C07K 2319/32 : fusions with soluble part of a cell surface receptor, "decoy receptors"
• CPC Class Codes: C07K 2319/90 : containing a motif for post-translational modification
• CPC Class Codes: C12N 2740/13043 : viral genome or elements thereof as genetic vector

Abstract

Disclosed herein are chimeric receptors comprising an extracellular domain with affinity and specific for the Fc portion of an immunoglobulin molecule (Ig) (e.g., an extracellular ligand-binding domain of F158 FCGR3A or V158 FCGR3A variant); a transmembrane domain (e.g., a transmembrane domain of CD8α); at least one co-stimulatory signaling domain (e.g., a co-stimulatory signaling domain of 4-1BB); and a cytoplasmic signaling domain comprising an immunoreceptor tyrosine-based activation motif (ITAM) (e.g., a cytoplasmic signaling domain of CD3ζ). Also provided herein are nucleic acids encoding such chimeric receptors and immune cells expressing the chimeric receptors. Such immune cells can be used to enhance antibody-dependent cell-mediated cytotoxicity and/or to enhance antibody-based immunotherapy, such as cancer immunotherapy.

Claims

1. A chimeric receptor, comprising:

(a) an extracellular ligand-binding domain of an Fc receptor;

(b) a transmembrane domain;

(c) at least one co-stimulatory signaling domain; and

(d) a cytoplasmic signaling domain comprising an immunoreceptor tyrosine-based activation motif (ITAM);

wherein either (c) or (d) is located at the C-terminus of the chimeric receptor; and

wherein if (a) is an extracellular ligand-binding domain of CD16A, (d) does not comprise an ITAM domain of an Fc receptor.

2. The chimeric receptor of claim 1, wherein (d) is located at the C-terminus of the chimeric receptor.

3. The chimeric receptor of claim 1, further comprising (e) a hinge domain which is located at the C-terminus of (a) and the N-terminus of (b).

4. The chimeric receptor of claim 1, wherein the chimeric receptor comprises:

(a) an extracellular ligand-binding domain of F158 FCGR3A or V158 FCGR3A variant,

(b) a hinge and transmembrane domain of CD8α

,

(c) a co-stimulatory signaling domain of 4-1BB, and

(d) a cytoplasmic signaling domain of CD3ζ

.

5. The chimeric receptor of claim 4, wherein the chimeric receptor further comprises a signal peptide of CD8α

.

6. The chimeric receptor of claim 4, wherein the extracellular domain of F158 FCGR3A and V158 FCGR3A variant consist of the amino acid sequences of SEQ ID NO:

70 and SEQ ID NO;

57, respectively.

7. The chimeric receptor of claim 4, wherein the hinge and transmembrane domains of CD8α

consist of the amino acid sequence of SEQ ID NO;

58.

8. The chimeric receptor of claim 4, wherein the co-stimulatory signaling domain of 4-1BB consists of the amino acid sequence of SEQ ID NO:

59.

9. The chimeric receptor of claim 4, wherein the cytoplasmic signaling domain of CD3ζ

consists of the amino acid sequence of SEQ ID NO;

60.

10. The chimeric receptor of claim 5, wherein the signal peptide of CD8α

consists of the amino acid sequence of SEQ ID NO;

61.

11. The chimeric receptor of claim 4, which comprises the amino acid sequence of residues 22 to 436 of SEQ ID NO:

1, or residues 22 to 436 of SEQ ID NO;

31.

12. The chimeric receptor of claim 5, which comprises the amino acid sequence of SEQ ID NO:

1 or SEQ ID NO;

31.

13. A nucleic acid comprising a nucleotide sequence encoding a chimeric receptor of claim 1.

14. The nucleic acid of claim 13, wherein the nucleotide sequence encodes a chimeric receptor that comprises:

(a) an extracellular ligand-binding domain of F158 FCGR3A or V158 FCGR3A variant,

(b) a hinge and transmembrane domain of CD8α

,

(c) a co-stimulatory signaling domain of 4-1BB, and

(d) a cytoplasmic signaling domain of CD3ζ

.

15. The nucleic acid of claim 14, wherein the nucleotide sequence encodes the amino acid sequence of residues 22 to 436 of SEQ ID NO:

1, or residues 22 to 436 of SEQ ID NO;

31.

16. The nucleic acid of claim 15, wherein the nucleotide sequence encodes the amino acid sequence of SEQ ID NO:

1 or SEQ ID NO;

31.

17. The nucleic acid of claim 16, wherein the nucleotide sequence comprises SEQ ID NO:

64 or SEQ ID NO;

71.

18. A pharmaceutical composition comprising the nucleic acid of claim 13 and a pharmaceutically acceptable carrier or excipient.

19. The pharmaceutical composition of claim 18, wherein the nucleic acid comprises a nucleotide sequence encoding a chimeric receptor comprising:

(a) an extracellular ligand-binding domain of F158 FCGR3A or V158 FCGR3A variant,

(b) a hinge and transmembrane domain of CD8α

,

(c) a co-stimulatory signaling domain of 4-1BB, and

(d) a cytoplasmic signaling domain of CD3ζ

.

20. A vector comprising the nucleic acid of claim 13.

21. The vector of claim 20, wherein the nucleic acid comprises a nucleotide sequence encoding a chimeric receptor comprising:

(a) an extracellular ligand-binding domain of F158 FCGR3A or V158 FCGR3A variant,

(b) a hinge and transmembrane domain of CD8α

,

(c) a co-stimulatory signaling domain of 4-1BB, and

(d) a cytoplasmic signaling domain of CD3ζ

.

22. The vector of claim 20, which is a viral vector.

23. The vector of claim 22, wherein the viral vector is a retroviral vector or a lentiviral vector.

24. A pharmaceutical composition comprising the vector of claim 20 and a pharmaceutically acceptable carrier.

25. A host cell comprising the chimeric receptor of claim 1.

26. The host cell of claim 25, wherein the chimeric receptor comprises:

(a) an extracellular ligand-binding domain of F158 FCGR3A or V158 FCGR3A variant,

(b) a hinge and transmembrane domain of CD8α

,

(c) a co-stimulatory signaling domain of 4-1BB, and

(d) a cytoplasmic signaling domain of CD3ζ

.

27. The host cell of claim 25, which is a T lymphocyte or an NK cell.

28. The host cell of claim 27, wherein the T lymphocyte or the NK cell is activated and/or expanded ex vivo.

29. The host cell of claim 27, wherein the T lymphocyte or NK cell is an autologous T lymphocyte or an autologous NK cell isolated from a patient having a cancer.

30. The host cell of claim 27, wherein the T lymphocyte or NK cell is an allogenic T lymphocyte or an allogenic NK cell.

31. The host cell of claim 30, wherein the T lymphocyte is an allogeneic T lymphocyte, in which the expression of the endogenous T cell receptor has been inhibited or eliminated.

32. The host cell of claim 28, wherein the T lymphocyte is activated in the presence of one or more agents selected from the group consisting of anti-CD3/CD28, IL-2, and phytohemoagglutinin.

33. The host cell of claim 28, wherein the NK cell is activated in the presence of one or more agents selected from the group consisting of CD 137 ligand protein, CD 137 antibody, IL-15 protein, IL-15 receptor antibody, IL-2 protein, IL-12 protein, IL-21 protein, and K562 cell line.

34. A pharmaceutical composition comprising the host cell of claim 25 and a pharmaceutically acceptable carrier or excipient.

35. The pharmaceutical composition of claim 34, further comprising an antibody that is cytotoxic to cancer cells.

36. The pharmaceutical composition of claim 35, wherein the antibody binds to cancer cells and has a human or humanized Fc portion which binds to human CD 16.

37. The pharmaceutical composition of claim 36, wherein the antibody is selected from the group consisting of Rituximab, Trastuzumab, hu14.18K322A, Epratuzumab, Cetuximab, and Labetuzumab.

38. A method for enhancing efficacy of an antibody-based immunotherapy of a cancer in a subject being treated with an anti-cancer antibody, the method comprising administering to the subject a therapeutically effective amount of T lymphocytes or NK cells that express the chimeric receptor of claim 1.

39. The method of claim 38, wherein the chimeric receptor comprises:

(a) an extracellular ligand-binding domain of F158 FCGR3A or V158 FCGR3A variant,

(b) a hinge and transmembrane domain of CD8α

,

(c) a co-stimulatory signaling domain of 4-1BB, and

(d) a cytoplasmic signaling domain of CD3ζ

.

40. The method of claim 38, wherein the antibody has a humanized Fc portion, which bind to human CD16.

41. The method of claim 38, wherein the cancer is selected from the group consisting of carcinoma, lymphoma, sarcoma, blastomas, and leukemia.

42. The method of claim 41, wherein the cancer is selected from the group consisting of a cancer of B-cell origin, breast cancer, gastric cancer, neuroblastoma, osteosarcoma, lung cancer, melanoma, prostate cancer, colon cancer, renal cell carcinoma, ovarian cancer, rhabdomyo sarcoma, leukemia, and Hodgkin'"'"'s lymphoma.

43. The method of claim 41, wherein the cancer of B-cell origin is selected from the group consisting of B-lineage acute lymphoblastic leukemia, B-cell chronic lymphocytic leukemia, and B-cell non-Hodgkin'"'"'s lymphoma.

44. The method of claim 38, wherein the antibody is selected from the group consisting of Rituximab, Trastuzumab, hu14.18K322A, Epratuzumab, Cetuximab, and Labetuzumab.

45. The method of claim 38, wherein the T lymphocytes or NK cells are autologous T lymphocytes or autologous NK cells isolated from the subject.

46. The method of claim 45, wherein, prior to the administration step, the autologous T lymphocytes or autologous NK cells are activated and/or expanded ex vivo.

47. The method of claim 38, wherein the T lymphocytes or NK cells are allogeneic T lymphocytes or allogeneic NK cells.

48. The method of claim 47, wherein the allogeneic T lymphocytes are T lymphocytes, in which the expression of the endogenous T cell receptor has been inhibited or eliminated.

49. The method of claim 47, wherein, prior to the administration step, the allogeneic T lymphocytes or allogeneic NK cells are activated and/or expanded ex vivo.

50. The method of claim 38, wherein the chimeric receptor is introduced into the T lymphocytes or the NK cells by a method selected from the group consisting of retroviral transduction, lentiviral transduction, DNA electroporation, and RNA electroporation.

51. A method of enhancing a T lymphocyte or an NK cell antibody-dependent cell cytotoxicity (ADCC) in a subject, the method comprising administering to the subject a therapeutically effective amount of T lymphocytes or NK cells that express the chimeric receptor of claim 1.

52. The method of claim 51, wherein the chimeric receptor comprising:

(a) an extracellular ligand-binding domain of F158 FCGR3A or V158 FCGR3A variant,

(b) the hinge and transmembrane domain of CD8a,

(c) a co-stimulatory signaling domain of 4-1BB, and

(d) a cytoplasmic signaling domain of CD3ζ

.

53. The method of claim 51, wherein the subject is being treated with an anti-cancer antibody.

54. The method of claim 51, wherein the antibody has a humanized Fc portion, which binds to human CD 16.

55. The method of claim 53, wherein the antibody is selected from the group consisting of Rituximab, Trastuzumab, hu14.18K322A, Epratuzumab, Cetuximab, and Labetuzumab.

56. The method of claim 53, wherein the subject has a cancer.

57. The method of claim 56, wherein the cancer is selected from the group consisting of carcinoma, lymphoma, sarcoma, blastoma, and leukemia.

58. The method of claim 57, wherein the cancer is selected from the group consisting of a cancer of B-cell origin, breast cancer, gastric cancer, neuroblastoma, osteosarcoma, lung cancer, melanoma, prostate cancer, colon cancer, renal cell carcinoma, ovarian cancer, rhabdomyo sarcoma, leukemia, and Hodgkin'"'"'s lymphoma.

59. The method of claim 58, wherein the cancer of B-cell origin is selected from the group consisting of B-lineage acute lymphoblastic leukemia, B-cell chronic lymphocytic leukemia, and B-cell non-Hodgkin'"'"'s lymphoma.

60. The method of claim 51, the T lymphocytes or NK cells are autologous T lymphocytes or autologous NK cells isolated from the subject.

61. The method of claim 60, wherein, prior to administering into the subject, the autologous T lymphocytes or autologous NK cells are activated and/or expanded ex vivo.

62. The method of claim 51, wherein the T lymphocytes or NK cells are allogeneic T lymphocytes or allogeneic NK cells.

63. The method of claim 62, wherein the allogeneic T lymphocytes are T lymphocytes in which the expression of the endogenous T cell receptor has been inhibited or eliminated.

64. The method of claim 63, wherein, prior to administering into the subject, the allogeneic T lymphocytes or allogeneic NK cells are activated and/or expanded ex vivo.

65. The method of claim 51, wherein the chimeric receptor is introduced into the T lymphocytes or the NK cells by a method selected from the group consisting of retroviral transduction, lentiviral transduction, DNA electroporation, and RNA electroporation.

66. The method of claim 51, wherein the T lymphocytes are activated in the presence of one or more agents selected from the group consisting of anti-CD3/CD28, IL-2, and phytohemoagglutinin.

67. The method of claim 51, wherein the NK cells are activated in the presence of one or more agents selected from the group consisting of CD137 ligand protein, CD137 antibody, IL-15 protein, IL-15 receptor antibody, IL-2 protein, IL-12 protein, IL-21 protein, and K562 cell line.

68. The method of claim 51, further comprising administering to the subject a therapeutically effective amount of IL-2.

69. A method for preparing immune cells expressing a chimeric receptor, comprising:

(i) providing a population of immune cells;

(ii) introducing into the immune cells a nucleic acid encoding a chimeric receptor of claim 1; and

(iii) culturing the immune cells under conditions allowing for expression of the chimeric receptor.

70. The method of claim 69, wherein the population of immune cells are derived from peripheral blood mononuclear cells (PBMC).

71. The method of claim 69, wherein the immune cells comprise natural killer cells or T lymphocytes.

72. The method of claim 69, wherein the immune cells are derived from a human cancer patient.

73. The method of claim 69, wherein the nucleic acid is a viral vector.

74. The method of claim 73, wherein the viral vector is a lentiviral vector or a retroviral vector.

75. The method of claim 69, wherein the vector is introduced into the immune cells by lentiviral transduction, retroviral transduction, DNA electroporation, or RNA electroporation.

76. The method of claim 69, further comprising (iv) activating the immune cells expressing the chimeric receptor.

77. The method of claim 71, wherein the immune cells comprise T cells, which are activated in the presence of one or more of anti-CD3 antibody, anti-CD28 antibody, IL-2, and phytohemoagglutinin.

78. The method of claim 77, wherein the immune cells comprise natural killer cells, which are activated in the presence of one or more of 4-1BB ligand, anti-4-1BB antibody, IL-15, anti-IL-15 receptor antibody, IL-2, IL-12, IL-21 and K562 cells.