Multivalent and Multispecific DR5-Binding Fusion Proteins

• US 20190309083A1
• Filed: 04/18/2019
• Published: 10/10/2019
• Est. Priority Date: 07/16/2015
• Status: Active Grant

• Current Assignee: Inhibrx, Inc.
• Original Assignee: Inhibrx, Inc.
• Inventors: Timmer, John C., Jones, Kyle S., Razai, Amir S., Hussain, Abrahim, Willis, Katelyn M., Deveraux, Quinn, Eckelman, Brendan P.
• Granted Patent: US 11,117,973 B2
• Time in Patent Office: Days
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 39/39558 against tumor tissues, cells, antigens
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: C07K 16/2878 against the NGF-receptor/TNF-receptor superfamily, e.g. CD27, CD30, CD40, CD95
• CPC Class Codes: C07K 2317/22 from camelids, e.g. camel, llama or dromedary
• CPC Class Codes: C07K 2317/24 containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
• CPC Class Codes: C07K 2317/33 Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
• CPC Class Codes: C07K 2317/35 Valency
• CPC Class Codes: C07K 2317/567 Framework region [FR]
• CPC Class Codes: C07K 2317/569 Single domain, e.g. dAb, sdAb, VHH, VNAR or nanobody®
• CPC Class Codes: C07K 2317/73 Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• CPC Class Codes: C07K 2317/75 Agonist effect on antigen

Abstract

The disclosure relates generally to molecules that specifically engage death receptor 5 (DR5), a member of the TNF receptor superfamily (TNFRSF). More specifically the disclosure relates to multivalent and multispecific molecules that bind at least DR5.

Claims

1. A method of treating a neoplasm comprising administering to a subject in need thereof a therapeutically effective amount of an isolated polypeptide that binds death receptor 5 (DR5) and comprises a plurality of DR5 binding domains (DR5BDs), wherein each DR5BD is a VHH comprising a CDR1 comprising the amino acid sequence of SEQ ID NO:

128, a CDR2 comprising the amino acid sequence of SEQ ID NO;

131, and a CDR3 comprising the amino acid sequence of SEQ ID NO;

130, and wherein adjacent DR5BDs are operably linked by an amino acid linker.

8. The method of claim 1, wherein the plurality of DR5BDs is two DR5BDs.

9. The method of claim 1, wherein the plurality of DR5BDs is four DR5BDs.

10. The method of claim 1, wherein the plurality of DR5BDs is six DR5BDs.

11. The method of claim 1, wherein each DR5BD comprises the amino acid sequence of SEQ ID NO:

87.

12. The method of claim 8, wherein each DR5BD comprises the amino acid sequence of SEQ ID NO:

87.

13. The method of claim 9, wherein each DR5BD comprises the amino acid sequence of SEQ ID NO:

87.

14. The method of claim 10, wherein each DR5BD comprises the amino acid sequence of SEQ ID NO:

87.

19. The method of claim 1, wherein the isolated polypeptide comprises an immunoglobulin hinge region and an immunoglobulin Fc region.

20. The method of claim 19, wherein the immunoglobulin Fc region is an IgG1 Fc region, an IgG2 Fc region, an IgG3 Fc region, or an IgG1 Fc region.

21. The method of claim 19, wherein the immunoglobulin Fc region comprises an amino acid sequence selected from SEQ ID NOs:

1-5 or 127.

26. The method of claim 1, wherein each VHH is a humanized VHH.

33. The method of claim 9, wherein the polypeptide is a homodimer of the structure:

DR5BD-Linker-DR5BD-Linker-Hinge-Fc, where each DR5BD is a humanized VHH sequence.

47. The method of claim 33, wherein each DR5BD comprises the amino acid sequence of SEQ ID NO:

87.

48. The method of claim 1, wherein the polypeptide comprises the amino acid sequence of SEQ ID NO:

113.

49. The method of claim 48, wherein the polypeptide is a homodimer of the amino acid sequence of SEQ ID NO:

113 fused to an Fc region polypeptide.

51. The method of claim 1, wherein each amino acid linker consists of 5-20 amino acids.

52. The method of claim 51, wherein each amino acid linker is composed predominantly of glycine and serine.

53. The method of claim 52, wherein each amino acid linker comprises an amino acid sequence selected from GGSGGS (SEQ ID NO:

11);

GGSGGSGGS (SEQ ID NO;

     12);

GGSGGSGGSGGS (SEQ ID NO;

     13); and

GGSGGSGGSGGSGGS (SEQ ID NO;

     14).

54. The method of claim 19, wherein the immunoglobulin hinge region comprises an amino acid sequence selected from EPKSSDKTHTCPPC (SEQ ID NO:

6), DKTHTCPPC (SEQ ID NO;

     7), and ESKYGPPCPPC (SEQ ID NO;

     8)

55. The method of claim 47, wherein each amino acid linker consists of 5-20 amino acids.

56. The method of claim 55, wherein each amino acid linker is composed predominantly of glycine and serine.

57. The method of claim 56, wherein each amino acid linker comprises an amino acid sequence selected from GGSGGS (SEQ ID NO:

11);

GGSGGSGGS (SEQ ID NO;

     12);

GGSGGSGGSGGS (SEQ ID NO;

     13); and

GGSGGSGGSGGSGGS (SEQ ID NO;

     14).

58. The method of claim 1, wherein the neoplasm is cancer.

59. The method of claim 9, wherein the neoplasm is cancer.

60. The method of claim 33, wherein the neoplasm is cancer.

61. The method of claim 48, wherein the neoplasm is cancer.

63. The method of claim 58, wherein the cancer is bladder cancer, breast cancer, uterine or cervical cancer, ovarian cancer, prostate cancer, testicular cancer, esophageal cancer, gastrointestinal cancer, pancreatic cancer, colorectal cancer, colon cancer, kidney cancer, head and neck cancer, lung cancer, stomach cancer, germ cell cancer, bone cancer, liver cancer, thyroid cancer, skin cancer, neoplasm of the central nervous system, lymphoma, leukemia, myeloma, sarcoma, mesothelioma, leukemia, lymphoma, myeloma, or virus-related cancer.

64. The method of claim 58, wherein the cancer is a metastatic cancer, refractory cancer, or recurrent cancer.

65. The method of claim 59, wherein the cancer is bladder cancer, breast cancer, uterine or cervical cancer, ovarian cancer, prostate cancer, testicular cancer, esophageal cancer, gastrointestinal cancer, pancreatic cancer, colorectal cancer, colon cancer, kidney cancer, head and neck cancer, lung cancer, stomach cancer, germ cell cancer, bone cancer, liver cancer, thyroid cancer, skin cancer, neoplasm of the central nervous system, lymphoma, leukemia, myeloma, sarcoma, mesothelioma, leukemia, lymphoma, myeloma, or virus-related cancer.

66. The method of claim 59, wherein the cancer is a metastatic cancer, refractory cancer, or recurrent cancer.

67. The method of claim 60, wherein the cancer is bladder cancer, breast cancer, uterine or cervical cancer, ovarian cancer, prostate cancer, testicular cancer, esophageal cancer, gastrointestinal cancer, pancreatic cancer, colorectal cancer, colon cancer, kidney cancer, head and neck cancer, lung cancer, stomach cancer, germ cell cancer, bone cancer, liver cancer, thyroid cancer, skin cancer, neoplasm of the central nervous system, lymphoma, leukemia, myeloma, sarcoma, mesothelioma, leukemia, lymphoma, myeloma, or virus-related cancer.

68. The method of claim 60, wherein the cancer is a metastatic cancer, refractory cancer, or recurrent cancer.

69. The method of claim 61, wherein the cancer is bladder cancer, breast cancer, uterine or cervical cancer, ovarian cancer, prostate cancer, testicular cancer, esophageal cancer, gastrointestinal cancer, pancreatic cancer, colorectal cancer, colon cancer, kidney cancer, head and neck cancer, lung cancer, stomach cancer, germ cell cancer, bone cancer, liver cancer, thyroid cancer, skin cancer, neoplasm of the central nervous system, lymphoma, leukemia, myeloma, sarcoma, mesothelioma, leukemia, lymphoma, myeloma, or virus-related cancer.

70. The method of claim 61, wherein the cancer is a metastatic cancer, refractory cancer, or recurrent cancer.

2-7. -7. (canceled)

15-18. -18. (canceled)

22-25. -25. (canceled)

27-32. -32. (canceled)

34-46. -46. (canceled)

50. A method of treating a neoplasm comprising administering to a subject in need thereof a therapeutically effective amount of an isolated polypeptide that binds death receptor 5 (DR5), wherein the polypeptide is a homodimer of the amino acid sequence of SEQ ID NO:

113 fused to an Fc region polypeptide of SEQ ID NO;

2.

62. The method of claim 50, wherein the neoplasm is cancer.

71. The method of claim 62, wherein the cancer is bladder cancer, breast cancer, uterine or cervical cancer, ovarian cancer, prostate cancer, testicular cancer, esophageal cancer, gastrointestinal cancer, pancreatic cancer, colorectal cancer, colon cancer, kidney cancer, head and neck cancer, lung cancer, stomach cancer, germ cell cancer, bone cancer, liver cancer, thyroid cancer, skin cancer, neoplasm of the central nervous system, lymphoma, leukemia, myeloma, sarcoma, mesothelioma, leukemia, lymphoma, myeloma, or virus-related cancer.

72. The method of claim 62, wherein the cancer is a metastatic cancer, refractory cancer, or recurrent cancer.

73. A method of modulating immune cells to enhance tumor destruction comprising administering to a subject in need thereof a therapeutically effective amount of an isolated polypeptide that binds at least death receptor 5 (DR5) and comprises a plurality of DR5 binding domains (DR5BDs), wherein each DR5BD is a VHH comprising a CDR1 comprising the amino acid sequence of SEQ ID NO:

128, a CDR2 comprising the amino acid sequence of SEQ ID NO;

131, and a CDR3 comprising the amino acid sequence of SEQ ID NO;

130, and wherein adjacent DR5BDs are operably linked by an amino acid linker.

74. A method of treating, alleviating a symptom of, ameliorating and/or delaying the progression of a viral, bacterial, or parasitic infection, comprising administering to a subject in need thereof a therapeutically effective amount of an isolated polypeptide that binds at least death receptor 5 (DR5) and comprises a plurality of DR5 binding domains (DR5BDs), wherein each DR5BD is a VHH comprising a CDR1 comprising the amino acid sequence of SEQ ID NO:

128, a CDR2 comprising the amino acid sequence of SEQ ID NO;

131, and a CDR3 comprising the amino acid sequence of SEQ ID NO;

130, and wherein adjacent DR5BDs are operably linked by an amino acid linker.

75. A method of treating, alleviating a symptom of, ameliorating and/or delaying the progression of an autoimmune disease or disorder, comprising administering to a subject in need thereof a therapeutically effective amount of an isolated polypeptide that binds at least death receptor 5 (DR5) and comprises a plurality of DR5 binding domains (DR5BDs), wherein each DR5BD is a VHH comprising a CDR1 comprising the amino acid sequence of SEQ ID NO:

128, a CDR2 comprising the amino acid sequence of SEQ ID NO;

131, and a CDR3 comprising the amino acid sequence of SEQ ID NO;

130, and wherein adjacent DR5BDs are operably linked by an amino acid linker.

76. A method of reducing or depleting the number of T regulatory cells in a tumor in a subject, comprising administering to the subject a therapeutically effective amount of an isolated polypeptide that binds at least death receptor 5 (DR5) and comprises a plurality of DR5 binding domains (DR5BDs), wherein each DR5BD is a VHH comprising a CDR1 comprising the amino acid sequence of SEQ ID NO:

128, a CDR2 comprising the amino acid sequence of SEQ ID NO;

131, and a CDR3 comprising the amino acid sequence of SEQ ID NO;

130, and wherein adjacent DR5BDs are operably linked by an amino acid linker.