Bispecific anti-Pseudomonas antibody having a modified Fc region and method of use thereof
Patent Information
- Authority / Receiving Office
- JP · JP
- Patent Type
- Applications
- Current Assignee / Owner
- ASTRAZENECA AB
- Filing Date
- 2024-05-08
- Publication Date
- 2026-06-09
Smart Images
Figure 2026518603000010 
Figure 2026518603000011 
Figure 2026518603000012
Abstract
Claims
1. A bispecific antibody that specifically binds to the Pseudomonas aeruginosa PcrV protein and Psl extracellular polysaccharide, wherein the antibody includes a modified IgG Fc region, and the modified IgG Fc region includes amino acid substitutions at two or more positions numbered according to the EU numbering index of Kabat, compared to the wild-type IgG Fc region, in this case, (i) Positions 432 and 437 are each replaced with cysteine, (ii) Position 433 is histidine, or is substituted with arginine, proline, threonine, lysine, serine, alanine, methionine, or asparagine. (iii) Position 434 is either asparagine or substituted with arginine, tryptophan, histidine, phenylalanine, tyrosine, serine, methionine, or threonine. (iv) Position 435 is histidine, and (v) Position 436 is tyrosine or phenylalanine, or is substituted with leucine, arginine, isoleucine, lysine, methionine, valine, histidine, serine, or threonine. The antibody is a bispecific antibody having an extended half-life compared to the corresponding antibody having a wild-type IgG Fc region.
2. The bispecific antibody according to claim 1, wherein the modified IgG Fc region is a modified IgG1 Fc region.
3. The bispecific antibody according to claim 1 or 2, wherein the modified IgG Fc region is a modified human IgG Fc region.
4. The bispecific antibody according to any one of claims 1 to 3, wherein the bispecific antibody aggregates less in solution than an antibody comprising a heavy chain having the amino acid sequence of SEQ ID NO: 19 and a light chain having the amino acid sequence of SEQ ID NO:
20.
5. The bispecific antibody according to any one of claims 1 to 4, wherein the bispecific antibody promotes the opsonin phagocytic death activity of P. aeruginosa, and optionally, the bispecific antibody mediates the in vitro opsonin phagocytic death activity of P. aeruginosa, similar to an antibody comprising a heavy chain containing the amino acid sequence of SEQ ID NO: 19 and a light chain containing the amino acid sequence of SEQ ID NO:
20.
6. A bispecific antibody according to any one of claims 1 to 5, further comprising an amino acid insertion after position 437, wherein the amino acid insertion is optionally glutamic acid.
7. The bispecific antibody according to any one of claims 1 to 6, wherein the binding affinity of the bispecific antibody to FcRn at pH 6.0 is higher than the binding affinity of the corresponding antibody having a wild-type human IgG1 Fc region to FcRn at pH 6.
8. The bispecific antibody according to any one of claims 1 to 7, wherein the binding affinity of the bispecific antibody to FcRn at pH 7.4 is higher than the binding affinity of the corresponding antibody having a wild-type human IgG1 Fc region to FcRn at pH 7.
4.
9. The bispecific antibody according to any one of claims 1 to 8, wherein the modified human IgG1 Fc region exhibits increased pH dependence for binding affinity to FcRn compared to the corresponding antibody having a wild-type human IgG1 Fc region.
10. The bispecific antibody according to any one of claims 1 to 9, wherein the modified human IgG1 Fc region has amino acid substitutions at three of the positions 432, 433, 434, 435, 436, and 437.
11. The bispecific antibody according to any one of claims 1 to 9, wherein the modified human IgG1 Fc region has amino acid substitutions at four of the positions 432, 433, 434, 435, 436, and 437.
12. The bispecific antibody according to any one of claims 1 to 9, wherein the modified human IgG1 Fc region has amino acid substitutions at five of the positions 432, 433, 434, 435, 436, and 437.
13. The bispecific antibody according to any one of claims 1 to 9, wherein the modified human IgG1 Fc region has amino acid substitutions at six of the positions 432, 433, 434, 435, 436, and 437.
14. The bispecific antibody according to any one of claims 1 to 9, wherein the modified human IgG1 Fc region comprises the amino acid sequence of SEQ ID NO: 44 or the amino acid sequence of SEQ ID NO:
33.
15. The bispecific antibody according to any one of claims 1 to 14, wherein the bispecific antibody is not a HexaBody.
16. The bispecific antibody according to any one of claims 1 to 15, wherein the bispecific antibody competitively inhibits the binding of the antibody to PcrV, the antibody comprising a heavy chain variable region (VH) containing the amino acid sequence of SEQ ID NO: 13 and a light chain variable region (VL) containing the amino acid sequence of SEQ ID NO:
14.
17. The bispecific antibody according to any one of claims 1 to 16, wherein the bispecific antibody binds to the same PcrV epitope as the antibody comprising VH containing the amino acid sequence of SEQ ID NO: 13 and VL containing the amino acid sequence of SEQ ID NO:
14.
18. The bispecific antibody according to any one of claims 1 to 17, wherein the bispecific antibody comprises an antigen-binding domain that binds to the Pseudomonas aeruginosa PcrV protein, and includes VH-CDR1 containing the amino acid sequence of SEQ ID NO: 1, VH-CDR2 containing the amino acid sequence of SEQ ID NO: 2, VH-CDR3 containing the amino acid sequence of SEQ ID NO: 3, VL-CDR1 containing the amino acid sequence of SEQ ID NO: 4, VL-CDR2 containing the amino acid sequence of SEQ ID NO: 5, and VL-CDR3 containing the amino acid sequence of SEQ ID NO:
6.
19. The bispecific antibody according to claim 18, wherein the antigen-binding domain that binds to the Pseudomonas aeruginosa PcrV protein comprises VH containing the amino acid sequence of SEQ ID NO: 13 and / or VL containing the amino acid sequence of SEQ ID NO:
14.
20. The bispecific antibody according to claim 18 or 19, wherein the antigen-binding domain that binds to the Pseudomonas aeruginosa PcrV protein comprises a heavy chain variable region and a light chain variable region on separate polypeptides.
21. The bispecific antibody according to any one of claims 1 to 20, wherein the bispecific antibody competitively inhibits the binding of an antibody comprising VH containing the amino acid sequence of SEQ ID NO: 15 and VL containing the amino acid sequence of SEQ ID NO: 16 to Psl.
22. The bispecific antibody according to any one of claims 1 to 21, wherein the bispecific antibody binds to the same Psl epitope as the antibody comprising VH containing the amino acid sequence of SEQ ID NO: 15 and VL containing the amino acid sequence of SEQ ID NO:
16.
23. The bispecific antibody according to any one of claims 1 to 22, wherein the antibody comprises an antigen-binding domain that binds to an extracellular polysaccharide of Pseudomonas aeruginosa Psl, and includes a heavy chain variable region VH-CDR1 containing the amino acid sequence of SEQ ID NO: 7, VH-CDR2 containing the amino acid sequence of SEQ ID NO: 8, VH-CDR3 containing the amino acid sequence of SEQ ID NO: 9, a light chain variable region VL-CDR1 containing the amino acid sequence of SEQ ID NO: 10, VL-CDR2 containing the amino acid sequence of SEQ ID NO: 11, and VL-CDR3 containing the amino acid sequence of SEQ ID NO:
12.
24. The bispecific antibody according to claim 23, wherein the antigen-binding domain that binds to the extracellular polysaccharide of Pseudomonas aeruginosa Psl comprises VH containing the amino acid sequence of SEQ ID NO: 15 and / or VL containing the amino acid sequence of SEQ ID NO:
16.
25. The bispecific antibody according to claim 23 or 24, wherein the antigen-binding domain that binds to the extracellular polysaccharide of Pseudomonas aeruginosa Psl comprises VH and VL on the same polypeptide.
26. The bispecific antibody according to any one of claims 23 to 25, wherein the antigen-binding domain that binds to the extracellular polysaccharide of Pseudomonas aeruginosa Psl comprises a linker between VH and VL, and optionally the linker comprises the amino acid sequence of SEQ ID NO:
18.
27. The bispecific antibody according to any one of claims 23 to 26, wherein the antigen-binding domain that binds to the extracellular polysaccharide of Pseudomonas aeruginosa Psl comprises scFv.
28. The bispecific antibody according to claim 27, wherein the scFv comprises a linker, and optionally the linker comprises the amino acid sequence of SEQ ID NO:
18.
29. The bispecific antibody according to claim 27 or claim 28, wherein the scFv is oriented in the VH-linker-VL configuration.
30. The bispecific antibody according to any one of claims 27 to 29, wherein the scFv comprises the amino acid sequence of SEQ ID NO:
17.
31. The bispecific antibody according to any one of claims 27 to 30, wherein the scFv is located on the same polypeptide chain as the VH of the antigen-binding domain that binds to the Pseudomonas aeruginosa PcrV protein.
32. The bispecific antibody according to any one of claims 27 to 31, wherein the scFv is C-terminal to the VH of the antigen-binding domain that binds to the Pseudomonas aeruginosa PcrV protein.
33. The bispecific antibody comprises (i) a heavy chain of formula VH-CH1-H1-L1-S-L2-H2-CH2-CH3, where VH is the anti-Pseudomonas aeruginosa PcrV heavy chain variable domain, CH1 is the heavy chain constant region domain 1, H1 is the first heavy chain hinge region fragment, L1 is the first linker, S is the anti-Pseudomonas aeruginosa Psl scFv molecule, L2 is the second linker, H2 is the second heavy chain hinge region fragment, CH2 is the heavy chain constant region domain-2, and CH3 is the heavy chain constant region domain-3, and (ii) a light chain of formula VL-CL, where VL is the anti-Pseudomonas A bispecific antibody according to any one of claims 1 to 32, comprising a light chain which is an aeruginosa PcrV light chain variable domain, wherein CL is the antibody light chain kappa constant region or the antibody light chain lambda constant region.
34. The bispecific antibody according to claim 33, wherein CH1 contains the amino acid sequence of SEQ ID NO:
21.
35. A bispecific antibody according to claim 33 or claim 34, wherein H1 contains the amino acid sequence of SEQ ID NO:
22.
36. A bispecific antibody according to any one of claims 33 to 35, wherein L1 contains the amino acid sequence of SEQ ID NO:
28.
37. A bispecific antibody according to any one of claims 33 to 36, wherein L2 contains the amino acid sequence of SEQ ID NO:
28.
38. A bispecific antibody according to any one of claims 33 to 37, wherein H2 contains the amino acid sequence of SEQ ID NO:
23.
39. A bispecific antibody according to any one of claims 33 to 38, wherein CH2-CH3 contains the amino acid sequence of SEQ ID NO:
30.
40. A bispecific antibody according to any one of claims 33 to 39, wherein CL is the constant region of the antibody light chain kappa.
41. A bispecific antibody according to any one of claims 33 to 40, wherein CL comprises the amino acid sequence of SEQ ID NO:
24.
42. A bispecific antibody according to any one of claims 1 to 41, comprising a heavy chain containing the amino acid sequence of SEQ ID NO: 31 and / or a light chain containing the amino acid sequence of SEQ ID NO:
20.
43. An isolated polynucleotide comprising a nucleic acid molecule encoding the heavy chain of a bispecific antibody according to any one of claims 1 to 42.
44. The isolated polynucleotide according to claim 43, further comprising a nucleic acid molecule encoding the light chain of a bispecific antibody according to any one of claims 1 to 42.
45. A vector comprising the polynucleotide described in claim 43 or 44.
46. (i) a host cell comprising the polynucleotide described in claim 43 or 44, or the vector described in claim 45.
47. A method for producing a bispecific antibody, comprising culturing the host cells described in claim 46, and optionally isolating the bispecific antibody.
48. A bispecific antibody produced by the method described in claim 47.
49. A composition comprising a bispecific antibody according to any one of claims 1 to 42 or 48, and a pharmaceutically acceptable carrier.
50. A method for treating or preventing a Pseudomonas infection in a subject requiring such treatment, comprising administering to the subject a bispecific antibody according to any one of claims 1 to 42 or 48, or a composition according to claim 49.
51. The method according to claim 50, wherein the infectious disease is a lung infection, respiratory tract infection, pneumonia, bacteremia, bone infection, joint infection, skin infection, burn infection, wound infection, or any combination thereof.
52. A method for treating bronchiectasis in a subject requiring treatment, comprising administering to the subject a bispecific antibody according to any one of claims 1 to 42 or 48, or a composition according to claim 49.
53. Forced expiratory vital capacity (FEV1) before bronchodilator administration in patients with bronchiectasis １ A method for improving ), comprising administering a bispecific antibody according to any one of claims 1 to 42 or 48, or a composition according to claim 49, to the subject.
54. A method for reducing the Pseudomonas aeruginosa load in a subject having bronchiectasis, comprising administering to the subject a bispecific antibody according to any one of claims 1 to 42 or 48, or a composition according to claim 49.
55. A method for reducing exacerbations of bronchiectasis in subjects, comprising administering to the subject a bispecific antibody according to any one of claims 1 to 42 or 48, or a composition according to claim 49.
56. A method for reducing the need for intravenous antibiotics in a subject with bronchiectasis, comprising administering to the subject a bispecific antibody according to any one of claims 1 to 42 or 48, or a composition according to claim 49.
57. A method for stabilizing lung function in a subject having bronchiectasis, comprising administering to the subject a bispecific antibody according to any one of claims 1 to 42 or 48, or a composition according to claim 49.
58. The method according to any one of claims 52 to 57, wherein the bronchiectasis is non-cystic fibrotic bronchiectasis.
59. The method according to any one of claims 50 to 58, further comprising administering an antibiotic.
60. The method according to any one of claims 50 to 59, wherein the subject is in which Pseudomonas aeruginosa is established, and optionally, the airway of the subject is in which Pseudomonas aeruginosa is established.
61. Use of a bispecific antibody according to any one of claims 1 to 42 or 48, or the composition according to claim 49, in the preparation of a pharmaceutical for use in the method according to any one of claims 50 to 60.
62. A bispecific antibody according to any one of claims 1 to 42 or 48, or a composition according to claim 49, for use in the method according to any one of claims 50 to 60.