Multitarget-directed protein complex and method of use
Patent Information
- Authority / Receiving Office
- JP · JP
- Patent Type
- Applications
- Current Assignee / Owner
- FBD BIOLOGICS LTD
- Filing Date
- 2024-06-20
- Publication Date
- 2026-06-25
Smart Images
Figure 2026520976000002 
Figure 2026520976000003 
Figure 2026520976000004
Abstract
Claims
1. (a) Fc and, (b) TIGIT (a T cell immune receptor having an immunoglobulin domain and an ITIM domain) ligand-binding domain, (c) PD-L1 (programmed death ligand 1) binding domain and (d) CD47 binding domain and A protein complex containing [the specified ingredient].
2. The protein complex according to claim 1, wherein the TIGIT ligand-binding domain can bind to cells expressing a TIGIT ligand (e.g., PVR or nectin-2) (e.g., cancer cells) and / or can block the interaction between TIGIT and the TIGIT ligand.
3. The protein complex according to claims 1 to 2, wherein the TIGIT ligand-binding domain is or comprises a TIGIT extracellular domain.
4. The protein complex according to claim 3, wherein the TIGIT extracellular domain comprises an amino acid sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:
1.
5. The protein complex according to any one of claims 1 to 4, wherein the PD-L1 binding domain can bind to cells expressing PD-L1 (e.g., cancer cells) and / or can block the interaction between PD-1 (programmed cell death protein 1) and PD-L1.
6. The protein complex according to any one of claims 1 to 5, wherein the PD-L1 binding domain is or comprises a PD-1 extracellular domain.
7. The protein complex according to claim 6, wherein the PD-1 extracellular domain comprises an amino acid sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:
2.
8. The protein complex according to claim 6, wherein the PD-1 extracellular domain includes a sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:
36.
9. The protein complex according to claim 8, wherein the amino acid corresponding to S39 of SEQ ID NO: 36 is H.
10. The protein complex according to claim 8 or 9, wherein the PD-1 extracellular domain further comprises a PD-L1 surface interaction sequence, and the PD-L1 surface interaction sequence comprises an amino acid sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NOs. 37, 38, 39, or 40.
11. The protein complex according to any one of claims 1 to 6 and 8 to 10, wherein the PD-L1 binding domain comprises an amino acid sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:
17.
12. The protein complex according to claims 1 to 11, wherein the CD47-binding domain can bind to cells expressing CD47 (e.g., cancer cells) and / or can block the interaction between CD47 and signal regulatory protein α (SIRPα).
13. The protein complex according to any one of claims 1 to 12, wherein the CD47-binding domain is or comprises a SIRPα extracellular domain.
14. The protein complex according to claim 13, wherein the SIRPα extracellular domain includes a sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:
3.
15. The protein complex according to claim 14, wherein the SIRPα extracellular domain contains one or more amino acid mutations at positions corresponding to H24, I31, E54, G55, H56, and / or D73 of SEQ ID NO:
3.
16. The SIRPα extracellular domain is as follows: (a) The amino acid corresponding to H24 in SEQ ID NO: 3 is R. (b) The amino acid corresponding to I31 in SEQ ID NO: 3 is T. (c) The amino acid corresponding to E54 in SEQ ID NO: 3 is A. (d) The amino acid corresponding to G55 in SEQ ID NO: 3 is K. (e) The amino acid corresponding to H56 in SEQ ID NO: 3 is Q, and (f) The amino acid corresponding to D73 in SEQ ID NO: 3 is I. The protein complex according to claim 14 or 15, comprising one or more of the following.
17. The protein complex according to any one of claims 1 to 16, wherein the CD47-binding domain comprises an amino acid sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, or 100% identical to SEQ ID NO:
18.
18. The protein complex according to any one of claims 1 to 17, wherein the TIGIT ligand-binding domain is optionally linked to the N-terminus of the CH2 domain in the Fc via a hinge region.
19. The protein complex according to claim 18, wherein the PD-L1 binding domain is optionally linked to the N-terminus of the TIGIT ligand binding domain via a first linker peptide.
20. The protein complex according to claim 19, wherein the CD47 binding domain is optionally linked to the C-terminus of the CH3 domain in the Fc via a second linker peptide.
21. The protein complex according to claim 18, wherein the CD47 binding domain is optionally linked to the C-terminus of the CH3 domain in the Fc via a first linker peptide.
22. The protein complex according to claim 21, wherein the PD-L1 binding domain is optionally linked to the C-terminus of the CD47 binding domain via a second linker peptide.
23. The protein complex according to any one of claims 1 to 17, wherein the PD-L1 binding domain is optionally linked to the N-terminus of the CH2 domain in the Fc via a hinge region.
24. The protein complex according to claim 23, wherein the TIGIT ligand-binding domain is optionally linked to the N-terminus of the PD-L1-binding domain via a first linker peptide.
25. The protein complex according to claim 24, wherein the CD47 binding domain is optionally linked to the C-terminus of the CH3 domain in the Fc via a second linker peptide.
26. The protein complex according to any one of claims 1 to 17, wherein the CD47-binding domain is optionally linked to the N-terminus of the CH2 domain in the Fc via a hinge region.
27. The protein complex according to claim 26, wherein the PD-L1 binding domain is optionally linked to the N-terminus of the CD47 binding domain via a first linker peptide.
28. The protein complex according to claim 27, wherein the TIGIT ligand-binding domain is optionally linked to the C-terminus of the CH3 domain in Fc via a second linker peptide.
29. The protein complex according to any one of claims 1 to 28, wherein the Fc is human IgG1Fc.
30. The protein complex according to any one of claims 1 to 28, wherein the Fc is human IgG4Fc.
31. The aforementioned hinge region is Human IgG4 hinge region optionally containing the S228P mutation according to EU numbering. The protein complex according to any one of claims 18 to 28 and 30.
32. (a) A first polypeptide comprising, in the direction from the N-terminus to the C-terminus, a first PD-L1 binding domain, an optional first linker peptide, a first TIGIT ligand binding domain, an optional first hinge region, a first Fc region, an optional second linker peptide, and a first CD47 binding domain, (b) A second polypeptide comprising, in the direction from the N-terminus to the C-terminus, a second PD-L1 binding domain, an optional third linker peptide, a second TIGIT ligand binding domain, an optional second hinge region, a second Fc region, an optional fourth linker peptide, and a second CD47 binding domain. A protein complex containing [the specified ingredient].
33. The protein complex according to claim 32, wherein the first PD-L1 binding domain and / or the second PD-L1 binding domain includes a sequence that is at least 80%, 90%, 95%, or 100% identical to SEQ ID NO: 2 or 17.
34. The protein complex according to claim 32 or 33, wherein the first TIGIT ligand-binding domain and / or the second TIGIT ligand-binding domain comprises a sequence that is at least 80%, 90%, 95%, or 100% identical to SEQ ID NO:
1.
35. The protein complex according to claims 32 to 34, wherein the first CD47-binding domain and / or the second CD47-binding domain comprises a sequence that is at least 80%, 90%, 95%, or 100% identical to Sequence ID No. 3 or 18.
36. The protein complex according to any one of claims 32 to 35, wherein the first hinge region and / or the second hinge region include a sequence that is at least 80% identical to Sequence ID No.
8.
37. The protein complex according to any one of claims 32 to 36, wherein the first Fc region and / or the second Fc region includes a sequence that is at least 80% identical to sequence number 9.
38. The protein complex according to any one of claims 32 to 35, wherein the first hinge region and / or the second hinge region include a sequence that is at least 80% identical to Sequence ID No.
10.
39. The protein complex according to any one of claims 32 to 35 and 38, wherein the first Fc region and / or the second Fc region includes a sequence that is at least 80% identical to SEQ ID NO:
11.
40. The protein complex according to any one of claims 32 to 39, wherein the first linker peptide and / or the third linker peptide comprises a sequence that is at least 80% identical to SEQ ID NO:
4.
41. The protein complex according to any one of claims 32 to 40, wherein the second linker peptide and / or the fourth linker peptide comprises a sequence that is at least 80% identical to SEQ ID NO:
5.
42. The protein complex according to any one of claims 32 to 41, wherein the first polypeptide and / or the second polypeptide comprises a sequence that is at least 80%, 90%, 95%, or 100% identical to SEQ ID NO: 12, 13, 19, or 20.
43. (a) A first polypeptide comprising, in the direction from the N-terminus to the C-terminus, a first TIGIT ligand-binding domain, an optional first linker peptide, a first PD-L1-binding domain, an optional first hinge region, a first Fc region, an optional second linker peptide, and a first CD47-binding domain, (b) A second polypeptide comprising, in the direction from the N-terminus to the C-terminus, a second TIGIT ligand-binding domain, an optional third linker peptide, a second PD-L1 binding domain, an optional second hinge region, a second Fc region, an optional fourth linker peptide, and a second CD47 binding domain. A protein complex containing [the specified ingredient].
44. The protein complex according to claim 43, wherein the first TIGIT-binding domain and / or the second TIGIT-binding domain comprises a sequence that is at least 80%, 90%, 95%, or 100% identical to Sequence ID No.
1.
45. The protein complex according to claim 43 or 44, wherein the first PD-L1 binding domain and / or the second PD-L1 binding domain comprises a sequence that is at least 80%, 90%, 95%, or 100% identical to Sequence ID No. 2 or 17.
46. The protein complex according to any one of claims 43 to 45, wherein the first CD47-binding domain and / or the second CD47-binding domain comprises a sequence that is at least 80%, 90%, 95%, and 100% identical to Sequence ID No. 3 or 18.
47. The protein complex according to any one of claims 43 to 46, wherein the first hinge region and / or the second hinge region include a sequence that is at least 80% identical to Sequence ID No.
8.
48. The protein complex according to any one of claims 43 to 47, wherein the first Fc region and / or the second Fc region includes a sequence that is at least 80% identical to sequence number 9.
49. The protein complex according to any one of claims 43 to 46, wherein the first hinge region and / or the second hinge region include a sequence that is at least 80% identical to sequence number 10.
50. The protein complex according to any one of claims 43 to 46 and 49, wherein the first Fc region and / or the second Fc region includes a sequence that is at least 80% identical to SEQ ID NO:
11.
51. The protein complex according to any one of claims 43 to 50, wherein the first linker peptide and / or the third linker peptide comprises a sequence that is at least 80% identical to SEQ ID NO:
4.
52. The protein complex according to any one of claims 43 to 51, wherein the second linker peptide and / or the fourth linker peptide comprises a sequence that is at least 80% identical to SEQ ID NO:
5.
53. The protein complex according to any one of claims 43 to 52, wherein the first polypeptide and / or the second polypeptide comprises a sequence that is at least 80%, 90%, 95%, or 100% identical to SEQ ID NO: 14 or 21.
54. (a) A first polypeptide comprising, in the direction from the N-terminus to the C-terminus, a first TIGIT ligand-binding domain, an optional first hinge region, a first Fc region, an optional first linker peptide, a first CD47-binding domain, an optional second linker peptide, and a first PD-L1-binding domain, (b) A second polypeptide comprising, in the direction from the N-terminus to the C-terminus, a second TIGIT ligand-binding domain, an optional second hinge region, a second Fc region, an optional third linker peptide, a second CD47-binding domain, an optional fourth linker peptide, and a second PD-L1-binding domain. A protein complex containing [the specified ingredient].
55. The protein complex according to claim 54, wherein the first TIGIT-binding domain and / or the second TIGIT-binding domain comprises a sequence that is at least 80%, 90%, 95%, or 100% identical to Sequence ID No.
1.
56. The protein complex according to claim 54 or 55, wherein the first CD47-binding domain and / or the second CD47-binding domain comprises a sequence that is at least 80%, 90%, 95%, or 100% identical to Sequence ID No. 3 or 18.
57. The protein complex according to any one of claims 54 to 56, wherein the first PD-L1 binding domain and / or the second PD-L1 binding domain includes a sequence that is at least 80%, 90%, 95%, or 100% identical to SEQ ID NO: 2 or 17.
58. The protein complex according to any one of claims 54 to 57, wherein the first hinge region and / or the second hinge region includes a sequence that is at least 80% identical to Sequence ID No.
8.
59. The protein complex according to any one of claims 54 to 58, wherein the first Fc region and / or the second Fc region includes a sequence that is at least 80% identical to sequence number 9.
60. The protein complex according to any one of claims 54 to 57, wherein the first hinge region and / or the second hinge region includes a sequence that is at least 80% identical to Sequence ID No.
10.
61. The protein complex according to any one of claims 54 to 57 and 60, wherein the first Fc region and / or the second Fc region includes a sequence that is at least 80% identical to SEQ ID NO:
11.
62. The protein complex according to any one of claims 54 to 61, wherein the first linker peptide and / or the third linker peptide comprises a sequence that is at least 80% identical to SEQ ID NO:
5.
63. The protein complex according to any one of claims 54 to 62, wherein the second linker peptide and / or the fourth linker peptide comprises a sequence that is at least 80% identical to SEQ ID NO:
4.
64. The protein complex according to any one of claims 54 to 63, wherein the first polypeptide and / or the second polypeptide comprises a sequence that is at least 80%, 90%, 95%, or 100% identical to SEQ ID NO: 15 or 22.
65. (a) A first polypeptide comprising, in the direction from the N-terminus to the C-terminus, a first PD-L1 binding domain, an optional first linker peptide, a first CD47 binding domain, an optional first hinge region, a first Fc region, an optional second linker peptide, and a first TIGIT ligand binding domain, (b) A second polypeptide comprising, in the direction from the N-terminus to the C-terminus, a second PD-L1 binding domain, an optional third linker peptide, a second CD47 binding domain, an optional second hinge region, a second Fc region, an optional fourth linker peptide, and a second TIGIT ligand binding domain. A protein complex containing [the specified ingredient].
66. The protein complex according to claim 65, wherein the first TIGIT-binding domain and / or the second TIGIT-binding domain comprises a sequence that is at least 80%, 90%, 95%, or 100% identical to Sequence ID No.
1.
67. The protein complex according to claim 65 or 66, wherein the first CD47-binding domain and / or the second CD47-binding domain comprises a sequence that is at least 80%, 90%, 95%, or 100% identical to Sequence ID No. 3 or 18.
68. The protein complex according to any one of claims 65 to 67, wherein the first PD-L1 binding domain and / or the second PD-L1 binding domain includes a sequence that is at least 80%, 90%, 95%, or 100% identical to SEQ ID NO: 2 or 17.
69. The protein complex according to any one of claims 65 to 68, wherein the first hinge region and / or the second hinge region includes a sequence that is at least 80% identical to Sequence ID No.
8.
70. The protein complex according to any one of claims 65 to 69, wherein the first Fc region and / or the second Fc region includes a sequence that is at least 80% identical to sequence number 9.
71. The protein complex according to any one of claims 65 to 68, wherein the first hinge region and / or the second hinge region include a sequence that is at least 80% identical to Sequence ID No.
10.
72. The protein complex according to any one of claims 65 to 68 and 71, wherein the first Fc region and / or the second Fc region includes a sequence that is at least 80% identical to SEQ ID NO:
11.
73. The protein complex according to any one of claims 65 to 72, wherein the first linker peptide and / or the third linker peptide comprises a sequence that is at least 80% identical to SEQ ID NO:
4.
74. The protein complex according to any one of claims 65 to 73, wherein the second linker peptide and / or the fourth linker peptide comprises a sequence that is at least 80% identical to SEQ ID NO:
5.
75. The protein complex according to any one of claims 65 to 74, wherein the first polypeptide and / or the second polypeptide comprises a sequence that is at least 80%, 90%, 95%, or 100% identical to SEQ ID NO:
16.
76. Polynucleotide encoding the protein complex according to any one of claims 1 to 75 Nucleic acids, including
77. The nucleic acid according to claim 76, which is DNA (e.g., cDNA) or RNA (e.g., mRNA).
78. A vector comprising one or more nucleic acids according to claim 76 or 77.
79. A cell comprising the vector according to claim 78.
80. The cell according to claim 79, which is a CHO cell.
81. A cell comprising one or more nucleic acids according to claim 76 or 77.
82. A method for producing a protein complex, (a) Culturing the cells according to any one of claims 79 to 81 under conditions sufficient for the cells to produce the protein complex, (b) Collecting the protein complex produced by the cells Methods that include...
83. A protein complex according to any one of claims 1 to 75, covalently bound to a therapeutic agent. A protein conjugate containing [the specified ingredient].
84. The protein conjugate according to claim 83, wherein the therapeutic agent is a cytotoxic or cell proliferation inhibitory agent.
85. A method for treating a subject with cancer, Administering a therapeutically effective amount of a composition comprising the protein complex described in any one of claims 1 to 75 or the protein conjugate described in claim 83 or 84 to the subject. Methods that include...
86. The method according to claim 85, wherein the subject has cancer cells expressing PVR, nectin-2, CD47, and / or PD-L1.
87. The method according to claim 85 or 86, wherein the cancer is breast cancer, prostate cancer, non-small cell lung cancer, pancreatic cancer, diffuse large B-cell lymphoma, mesothelioma, lung cancer, ovarian cancer, colon cancer, pleural tumor, gliablastoma, esophageal cancer, gastric cancer, synovial sarcoma, thymic carcinoma, endometrial cancer, stomach cancer, bile duct cancer, head and neck cancer, hematological cancer, or a combination thereof.
88. A method to reduce the rate of tumor growth, The tumor cells are brought into contact with an effective amount of a composition comprising the protein complex described in any one of claims 1 to 75 or the protein conjugate described in claim 83 or 84. Methods that include...
89. A method to kill tumor cells, The tumor cells are brought into contact with an effective amount of a composition comprising the protein complex described in any one of claims 1 to 75 or the protein conjugate described in claim 83 or 84. Methods that include...
90. A pharmaceutical composition comprising a protein complex according to any one of claims 1 to 75 and a pharmaceutically acceptable carrier.