TGF-Beta Antagonist Multi-Target Binding Proteins

• US 20110177070A1
• Filed: 07/02/2009
• Published: 07/21/2011
• Est. Priority Date: 07/02/2008
• Status: Abandoned Application

• Current Assignee: Emergent Product Development Seattle, LLC
• Original Assignee: Emergent Product Development Seattle, LLC
• Inventors: Thompson, Peter Armstrong, Misher, Lynda, Baum, Peter Robert, Lofquist, Alan Keith
• Application Number:
  US13/001,088
• Publication Number:
  US 20110177070A1
• Time in Patent Office: Days
• US Class Current: 424/134.1
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C07K 14/71 for growth factors; for growth regulators
• CPC Class Codes: C07K 14/715 for cytokines; for lymphokines; for interferons
• CPC Class Codes: C07K 14/7151 for tumor necrosis factor [TNF], for lymphotoxin [LT]
• CPC Class Codes: C07K 14/7155 for interleukins [IL]
• CPC Class Codes: C07K 16/22 against growth factors ; against growth regulators
• CPC Class Codes: C07K 16/24 against cytokines, lymphokines or interferons
• CPC Class Codes: C07K 2317/55 Fab or Fab'
• 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 2319/32 fusions with soluble part of a cell surface receptor, "decoy receptors"
• CPC Class Codes: C07K 2319/74 containing a fusion for binding to a cell surface receptor

Abstract

This disclosure provides a multi-target fusion protein composed of a TGF? antagonist domain and another binding domain antagonistic for a heterologous target (such as IL6, IL10, VEGF, TNF, HGF, TWEAK, IGF) or agonistic for a heterologous target (such as GITR). The multi-specific fusion protein may also include an intervening domain that separates the binding domains and allows for dimerization. This disclosure also provides polynucleotides encoding the multi-specific fusion proteins, compositions of the fusion proteins, and methods of using the multi-specific fusion proteins and compositions.

Claims

1. A multi-specific fusion protein comprising a structure from amino terminus to carboxy terminus selected from the group consisting of:

(a) BD-ID-ED;

(b) ED-ID-BD; and

(c) ED1-ID-ED2

wherein;

ED is a TGFβ

antagonist and ED1 and ED2 are different antagonists wherein ED1 or ED2 is a TGFβ

antagonist;

ID is an intervening domain; and

BD is a binding domain of a TNF antagonist, an IL6 antagonist, an IL10 antagonist, a VEGF antagonist, an HGF antagonist, an IGF antagonist, or a GITR agonist.

2. The multi-specific fusion protein of claim 1, wherein the BD is an immunoglobulin variable binding domain.

3. The multi-specific fusion protein of claim 1, wherein the ED is a receptor ligand binding domain.

4. The multi-specific fusion protein of claim 1, wherein the intervening domain has the following structure:

-L1-CH2CH3-,

wherein;

L1 is an immunoglobulin hinge linker, and

—

CH2CH3- is the CH2CH3 region of an IgG1 Fc domain.

5. The multi-specific fusion protein of claim 1, wherein the BD is connected to the intervening domain by a first linker and the ED is connected to the intervening domain by a second linker and wherein the first and second linkers are the same or different.

6. The multi-specific fusion protein of claim 5, wherein the first and second linkers are selected from the group consisting of SEQ ID NO:

497-604 and 1223-122.

7. The multi-specific fusion protein of claim 1, comprising an amino acid sequence which is selected from the group consisting of SEQ ID NOS:

735-742.

10. A polynucleotide encoding the multi-specific fusion protein of claim 1.

11. An expression vector comprising the polynucleotide according to claim 10, which is operably linked to an expression control sequence.

12. A host cell comprising the expression vector according to claim 11.

13. A method for treating a subject with a malignant condition comprising administering to a subject in need thereof a therapeutically effective amount of the multi-specific fusion protein of claim 1.

14. The method of claim 13 wherein the malignant condition is selected from the group consisting of breast cancer, renal cell carcinoma, melanoma and prostate cancer.

15. The multi-specific fusion protein of claim 4, wherein said immunoglobulin hinge linker is an IgG1 hinge having the first cysteine substituted with a different amino acid.

16. The multi-specific fusion protein of claim 4, wherein said CH2CH3 region of an IgG1 Fc domain is mutated to eliminate Fcγ

RI-III interaction while retaining FcRn interaction.

17. The multi-specific fusion protein of claim 6, wherein the first linker is SEQ ID NO:

576 and the second linker is SEQ ID NO;

1223.

18. A method of producing a multi-specific fusion protein comprising culturing the host cell of claim 12 in a medium and expressing the protein.

8. A composition comprising the multi-specific fusion protein of claim and a pharmaceutically acceptable carrier, diluent, or excipient.

9. The composition of claim 8 wherein the multi-specific fusion protein exists as a dimer or a multimer in the composition.