FC FUSION PROTEINS COMPRISING NOVEL LINKERS OR ARRANGEMENTS

• US 20140113370A1
• Filed: 04/13/2012
• Published: 04/24/2014
• Est. Priority Date: 04/13/2011
• Status: Active Grant

• Current Assignee: Bristol-Myers Squibb Company
• Original Assignee: Bristol-Myers Squibb Company
• Inventors: Camphausen, Ray, Saeed-Kothe, Amna, Davis, Jonathan, Mitchell, Tracy S.
• Granted Patent: US 9,469,676 B2
• Time in Patent Office: Days
• US Class Current: 435/328
• 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: C07K 14/47 from mammals
• CPC Class Codes: C07K 14/78 Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
• CPC Class Codes: C07K 16/244 Interleukins [IL]
• CPC Class Codes: C07K 16/2863 against receptors for growth factors, growth regulators
• CPC Class Codes: C07K 16/40 against enzymes
• CPC Class Codes: C07K 16/46 Hybrid immunoglobulins hybrids of an immunoglobulin with a peptide not being an immunoglobulin C07K19/00
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C07K 2317/94 Stability, e.g. half-life, pH, temperature or enzyme-resistance
• CPC Class Codes: C07K 2318/20 Antigen-binding scaffold molecules wherein the scaffold is not an immunoglobulin variable region or antibody mimetics
• CPC Class Codes: C07K 2319/30 Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
• CPC Class Codes: C07K 7/06 having 5 to 11 amino acids
• CPC Class Codes: C07K 7/08 having 12 to 20 amino acids gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68
• CPC Class Codes: G01N 2333/948 acting on peptide bonds (3.4)
• CPC Class Codes: G01N 33/573 for enzymes or isoenzymes

Abstract

The application provides Fc fusion proteins having novel arrangements. In one embodiment, the application provides Fc fusion proteins comprising a ¹⁰Fn3 domain. In another embodiment, the application provides Fc fusion proteins comprising linkers derived from the naturally occurring C-terminal tail regions of membrane bound or secretory immunoglobulins.

Claims

1. A polypeptide comprising:

(a) a ¹⁰Fn3 domain having an altered amino acid sequence relative to the wild-type sequence, wherein the ¹⁰Fn3 domain binds to a target molecule with a K_D of less than 500 nM;

(b) an immunoglobulin (Ig) Fc domain; and

(c) a hinge sequence.

2. The polypeptide of claim 1, wherein the polypeptide has one of the following arrangements from N-terminus to C-terminus:

¹⁰Fn3 domain-hinge-Fc domain or hinge-Fc domain-linker-¹⁰Fn3 domain.

4. The polypeptide of claim 1, wherein the polypeptide is a dimer.

5. The polypeptide of claim 1, wherein the polypeptide further comprises a second ¹⁰Fn3 domain having an altered amino acid sequence relative to the wild-type sequence and wherein the second ¹⁰Fn3 domain binds to a target molecule with a K_D of less than 10 μ

M.

19. A nucleic acid encoding a polypeptide of claim 1.

20. A vector comprising the nucleic acid of claim 19.

22. A host cell comprising the vector of claim 20.

32. A method of detecting a target molecule in a sample, comprising:

(a) contacting the sample with the polypeptide of claim 1, wherein said contacting is carried out under conditions that allow the polypeptide to form a complex with the target molecule; and

(b) detecting the complex.

33. A method of treating a disease or disorder comprising administering the polypeptide of claim 1 to a subject in need thereof.

3. (canceled)

6-18. -18. (canceled)

21. (canceled)

23. A polypeptide comprising an immunoglobulin Fc domain and a heterologous polypeptide, wherein the heterologous polypeptide is fused to the N-terminus or the C-terminus of the Fc domain by a polypeptide linker, wherein the polypeptide linker comprises a sequence that is derived from the C-terminal tail region of the heavy chain of a membrane bound or secretory immunoglobulin or that is (a) at least 80% identical to any one of SEQ ID NOs:

33, 37, 38, 45, 47, 48, 50, 81-83, and 90-98, (b) comprises at least 5 contiguous amino acids of any one of SEQ ID NOs;

33, 37, 38, 45, 47, 48, 50, 81-83, and 90-98, (c) comprises at least 10 contiguous amino acids of any one of SEQ ID NOs;

33, 47, 48, 50, 81-83, and 90-98, or (d) comprises any one of SEQ ID NOs;

33, 37, 38, 45, 47, 48, 50, 81-83, and 90-98.

24. The polypeptide of claim 23, wherein the polypeptide linker comprises a sequence that is derived from the C-terminal tail region of the heavy chain of a membrane bound or secretory immunoglobulin, and the linker is (a) at least 80% identical to any one of SEQ ID NOs:

46, 49, 51-70, or 86 or, (b) comprises at least 5 contiguous amino acids of any one of SEQ ID NOs;

46, 49, 51-70, or 8651-70, (c) comprises at least 10 contiguous amino acids of any one of SEQ ID NOs;

46, 49, 51-70, or 86, or (d) comprises any one of SEQ ID NOs;

46, 49, 51-70, or 86.

25. The polypeptide of claim 23, wherein the heterologous polypeptide is fused to the C-terminus of the Fc domain.

26. The polypeptide of claim 23, wherein the heterologous polypeptide is fused to the N-terminus of the Fc domain.

27. The polypeptide of claim 23, wherein the heterologous polypeptide comprises a ¹⁰Fn3 domain.

28. The polypeptide of claim 23, wherein the immunoglobulin Fc domain comprises a hinge or a portion thereof.

29. A nucleic acid encoding a polypeptide of claim 23.

30. A vector comprising the nucleic acid of claim 29.

31. A host cell comprising the vector of claim 30.

34. A method of detecting a target molecule in a sample, comprising:

(a) contacting the sample with the polypeptide of claim 23, wherein said contacting is carried out under conditions that allow the polypeptide to form a complex with the target molecule; and

(b) detecting the complex.

35. A method of treating a disease or disorder comprising administering the polypeptide of claim 23 to a subject in need thereof.