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EP-4739703-A1 - HETERODIMERIC FC MOLECULES AND USES THEREOF

EP4739703A1EP 4739703 A1EP4739703 A1EP 4739703A1EP-4739703-A1

Abstract

The present disclosure provides heteromultimeric complexes comprising a first polypeptide and a second polypeptide, wherein the first and second polypeptides are not the same and comprise a heteromultimerization modification that reduces the formation of homomultimers and promotes the formation of heteromultimers. The present disclosure further provides fusion proteins, multispecific ligand-binding proteins, conjugates, and pharmaceutical compositions comprising such heteromultimeric complexes, including methods of making and uses to treat a wide range of diseases or conditions.

Inventors

  • IM, Sunkyoung
  • LEE, MINJI
  • CHOI, DONGHOON
  • YU, TAEKYUNG
  • LEE, JIE-OH
  • JEONG, HYE JIN
  • CHO, GEUN YOUNG

Assignees

  • NeoImmuneTech, Inc.
  • Postech Academy-Industry Foundation

Dates

Publication Date
20260513
Application Date
20240702

Claims (20)

  1. 1. A heterodimeric protein comprising a first polypeptide and a second polypeptide, wherein (a) the first polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 27, and (b) the second polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 32 or SEQ ID NO: 29.
  2. 2. The heterodimeric protein of claim 1, wherein the second polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 32. 3. The heterodimer protein of claim 1, wherein the second polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 29. 4. A heterodimeric protein comprising a first polypeptide and a second polypeptide, wherein (a) the first polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 16, and (b) the second polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 20. 5. A heterodimeric protein comprising a first polypeptide and a second polypeptide, wherein the first polypeptide and second polypeptide are not the same, wherein the first polypeptide comprises a first CH3 domain and the second polypeptide comprises a second CH3 domain, wherein (a) the first CH3 domain comprises the amino acid sequence set forth in SEQ ID NO: 24, and (b) the second CH3 domain comprises the amino acid sequence set forth in SEQ ID NO: 26, wherein: (1) amino acid residue N50, S60, or both of the first CH3 domain are not modified or modified with a positive charge modification, and amino acid residue N50, S60, or both of the second CH3 domain are not modified or modified with a negative charge modification, such that the first CH3 domain and the second CH3 domain have opposite charges; (2) amino acid residue N50, S60, or both of the first CH3 domain are not modified or modified with a negative charge modification, and amino acid residue N50, S60, or both of the second CH3 domain are not modified or modified with a positive charge modification, such that the first CH3 domain and the second CH3 domain have opposite charges;
  3. (3) amino acid residue D59, R69, or both of the first CH3 domain are not modified or modified with a positive charge modification, and amino acid residue D59, R69, or both of the second CH3 domain are not modified or modified with a negative charge modification, such that the first CH3 domain and the second CH3 domain have opposite charges;
  4. (4) amino acid residue D59, R69, or both of the first CH3 domain are not modified or modified with a negative charge modification, and amino acid residue D59, R69, or both of the second CH3 domain are not modified or modified with a positive charge modification, such that the first CH3 domain and the second CH3 domain have opposite charges;
  5. (5) amino acid residue K52 of the first CH3 domain is not modified or modified with a positive charge modification, and amino acid residue F65 of the second CH3 domain is not modified or modified with a negative charge modification, such that the first CH3 domain and the second CH3 domain have opposite charges; and/or (6) amino acid residue K52 of the first CH3 domain is not modified or modified with a negative charge modification, and amino acid residue F65 of the second CH3 domain is not modified or modified with a positive charge modification, such that the first CH3 domain and the second CH3 domain have opposite charges.
  6. 6. The heterodimeric protein of claim 5, wherein the modification in the first CH3 domain comprises (a) N50K or N50R, (b) K52R, (c) D59K or D59R, (d) S60K or S60R, (e) R69K, or (f) any combination thereof; and wherein the modification in the second CH3 comain comprises (a) N50D or N50E, (b) D59E, (c) S60D or S60E, (d) R69D or R69E, (e) F65D or F65E, or (f) any combination thereof.
  7. 7. The heterodimeric protein of claim 5, wherein the modification in the first CH3 domain comprises (a) N50D or N50E, (b) K52D or K52E, (c) D59E, (d) S60D or S60E, (e) R69D or R69E, or (f) any combination thereof; and wherein the modification in the second CH3 domain comprises (a) N50K or N50R, (b) D59K or D59R, (c) S60K or S60R, (d) R69K, (e) F65K or F65R, or (f) any combination thereof.
  8. 8. The heterodimeric protein of any one of claims 5 to 7, wherein: (a) the amino acid residue N50 of the first CH3 domain is modified to N50K or N50R and the amino acid residue S60 of the second CH3 domain is modified to S60D or S60E, (b) the amino acid residue S60 of the first CH3 domain is modified to S60D or S60E and the amino acid residue N50 of the second CH3 domain is modified to N50K or N50R, (c) the amino acid residue K52 of the first CH3 domain is not modified or modified to K52R and the amino acid residue F65 of the second CH3 domain is modified to F65D or F65E, (d) the amino acid residue D59 of the first CH3 domain is modified to D59K or D59R and the amino acid residue R69 of the second CH3 domain is modified to R69D or R69E, (e) the amino acid residue R69 of the first CH3 domain is modified R69D or R69E and the amino acid residue D59 of the second CH3 domain is modified D59K or D59R, or (f) any combination of (a) to (e).
  9. 9. The heterodimeric protein of any one of claims 5 to 8, wherein the first CH3 domain or the second CH3 domain comprises the amino acid sequence set forth in SEQ ID NO: 28, SEQ ID NO: 31, or SEQ ID NO: 33.
  10. 10. The heterodimeric protein of any one of claims 5 to 8, wherein the first CH3 domain comprises the amino acid sequence set forth in SEQ ID NO: 28, and the second CH3 domain comprises the amino acid sequence set forth in SEQ ID NO: 31 or SEQ ID NO: 33.
  11. 11. The heterodimeric protein of any one of claims 5 to 8, wherein: (a) the first polypeptide further comprises a first hinge domain, a first CH2 domain, or both; (b) the second polypeptide further comprises a second hinge domain, a second CH2 domain, or both; or (c) both (a) and (b).
  12. 12. The heterodimeric protein of claim 11, wherein the first hinge domain and the second hinge domain are each selected from an IgGl hinge, an IgG2 hinge, an IgG3 hinge, an IgG4 hinge, an IgD hinge, or a synthetic linker.
  13. 13. The heterodimeric protein of claim 12, wherein the synthetic linker comprises a glycineserine linker, a glycine-alanine linker, an alanine-serine linker, or combinations thereof.
  14. 14. The heterodimeric protein of any one of claims 11 to 13, wherein the first hinge domain and the second hinge domain are the same or different.
  15. 15. The heterodimeric protein of any one of claims 11 to 14, wherein: (a) the first hinge domain comprises the amino acid sequence set forth in SEQ ID NO: 22, (b) the second hinge domain comprises the amino acid sequence forth in SEQ ID NO: 22, or (c) both (a) and (b).
  16. 16. The heterodimeric protein of any one of claims 11 to 15, wherein the first CH2 domain and the second CH2 domain are the same or different.
  17. 17. The heterodimeric protein of claim 16, wherein: (a) the first CH2 domain comprises the amino acid sequence set forth in SEQ ID NO: 25, (b) the second CH2 domain comprises the amino acid sequence set forth in SEQ ID NO: 25, or (c) both (a) and (b).
  18. 18. The heterodimeric protein of any one of claims 1 to 17, further comprising a first biologically active molecule.
  19. 19. The heterodimeric protein of any one of claim 18, wherein the first biologically active molecule is attached to the N-terminus of the first polypeptide, the C-terminus of the first polypeptide, the N-terminus of the second polypeptide, the C-terminus of the second polypeptide, or any combination thereof.
  20. 20. The heterodimeric protein of any one of claim 19, wherein the first biologically active molecule is attached directly or via a linker.

Description

HETERODIMERIC FC MOLECULES AND USES THEREOF CROSS REFERENCE TO RELATED APPLICATIONS [0001] This PCT application claims the priority benefit of U.S. Provisional Application No. 63/511,726, filed July 3, 2023, which is incorporated herein by reference in its entirety. REFERENCE TO SEQUENCE LISTING SUBMITTED ELECTRONICALLY [0002] The content of the electronically submitted sequence listing in .XML file (4241_042PC01_SequenceListing_ST26.xml; 85,846 bytes; July 2, 2024) filed with the application is incorporated herein by reference in its entirety. FIELD OF THE DISCLOSURE [0003] The present disclosure provides heteromultimeric complexes (e.g., heterodimeric proteins) comprising a modified Fc and uses of such complexes. BACKGROUND OF THE DISCLOSURE [0004] Antibody-based therapeutics have been used successfully to treat a variety of diseases, including cancer and autoimmune/inflammatory disorders. Yet, improvements are still needed, particularly with respect to enhancing their clinical efficacy. The development of multispecific antibodies (e.g., bispecific antibodies) has great clinical potential (e.g., can recognize two different antigens, recruit different types of effector cells, and/or modulate multiple signaling pathways). However, production of multispecific antibodies can be challenging. The broad application of multispecific antibodies has been hindered by the difficulties of developing a platform for producing such antibodies that exhibit favorable half-life, high stability, lack of immunogenicity, and feasibilities for large scale manufacturing and purification. Therefore, there remains a need for a new and improved platform for developing heteromultimeric complexes (e.g., multispecific antibodies) that can be used in treating a wide range of diseases and conditions. BRIEF SUMMARY OF THE DISCLOSURE [0005] Provided herein is a heterodimeric protein comprising a first polypeptide and a second polypeptide, wherein (a) the first polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 16, and (b) the second polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 20. [0006] Also provided herein is a heterodimeric protein comprising a first polypeptide and a second polypeptide, wherein (a) the first polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 27, and (b) the second polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 29 or SEQ ID NO: 32. In some aspects, the second polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 29. In some aspects, the second polypeptide comprises the amino acid sequence set forth in SEQ ID NO: 32. [0007] Some aspects of the present disclosure is related to a heterodimeric protein comprising a first polypeptide and a second polypeptide, wherein the first polypeptide and second polypeptide are not the same, wherein the first polypeptide comprises a first CH3 domain and the second polypeptide comprises a second CH3 domain, wherein (a) the first CH3 domain comprises the amino acid sequence set forth in SEQ ID NO: 24, and (b) the second CH3 domain comprises the amino acid sequence set forth in SEQ ID NO: 26, wherein: (1) amino acid residue N50, S60, or both of the first CH3 domain are not modified or modified with a positive charge modification, and amino acid residue N50, S60, or both of the second CH3 domain are not modified or modified with a negative charge modification, such that the first CH3 domain and the second CH3 domain have opposite charges; (2) amino acid residue N50, S60, or both of the first CH3 domain are not modified or modified with a negative charge modification, and amino acid residue N50, S60, or both of the second CH3 domain are not modified or modified with a positive charge modification, such that the first CH3 domain and the second CH3 domain have opposite charges; (3) amino acid residue D59, R69, or both of the first CH3 domain are not modified or modified with a positive charge modification, and amino acid residue D59, R69, or both of the second CH3 domain are not modified or modified with a negative charge modification, such that the first CH3 domain and the second CH3 domain have opposite charges; (4) amino acid residue D59, R69, or both of the first CH3 domain are not modified or modified with a negative charge modification, and amino acid residue D59, R69, or both of the second CH3 domain are not modified or modified with a positive charge modification, such that the first CH3 domain and the second CH3 domain have opposite charges; (5) amino acid residue K52 of the first CH3 domain is not modified or modified with a positive charge modification, and amino acid residue F65 of the second CH3 domain is not modified or modified with a negative charge modification, such that the first CH3 domain and the second CH3 domain have opposite charges; and/or (6) amino acid residue K52 of the first CH3 domain is not modified or modified with a negative charge modification, and amino