Variants derived from ActRIIB

• US 9,399,669 B2
• Filed: 12/28/2012
• Issued: 07/26/2016
• Est. Priority Date: 02/02/2007
• Status: Active Grant

• Current Assignee: Acceleron Pharma Inc. (Merck & Co., Inc.)
• Original Assignee: Acceleron Pharma Inc. (Merck & Co., Inc.)
• Inventors: Knopf, John, Seehra, Jasbir, Kumar, Ravindra
• Primary Examiner(s): KEMMERER, ELIZABETH
• Application Number:
  US13/730,418
• Publication Number:
  US 20130184210A1
• Time in Patent Office: 1,306 Days
• Field of Search: None
• US Class Current: 1/1
• 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: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 11/00 Drugs for disorders of the respiratory system
• CPC Class Codes: A61P 13/08 of the prostate
• CPC Class Codes: A61P 19/02 for joint disorders, e.g. arthritis, arthrosis
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 21/02 Muscle relaxants, e.g. for tetanus or cramps
• CPC Class Codes: A61P 21/04 for myasthenia gravis
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 3/00 Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/04 Anorexiants; Antiobesity agents
• CPC Class Codes: A61P 3/06 Antihyperlipidemics
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/12 Antihypertensives
• CPC Class Codes: C07K 14/705 : Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: C07K 14/71 : for growth factors; for growth regulators
• CPC Class Codes: C07K 14/72 : for hormones for neuromediators C07K14/70571
• CPC Class Codes: C07K 16/18 : against material from animals or humans
• 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 2319/70 : containing domain for protein-protein interaction

Abstract

In certain aspects, the present invention provides compositions and methods for modulating (promoting or inhibiting) growth of a tissue, such as bone, cartilage, muscle, fat, and/or neuronal tissue. The present invention also provides methods of screening compounds that modulate activity of an ActRIIB protein and/or an ActRIIB ligand. The compositions and methods provided herein are useful in treating diseases associated with abnormal activity of an ActRIIB protein and/or an ActRIIB ligand.

Claims

1. A variant ActRIIB protein comprising an amino acid sequence that is at least 95% identical to the sequence of amino acids 29-109 of SEQ ID NO:

2, wherein the protein comprises at least one N-X-S/T sequence at a position other than an endogenous N-X-S/T sequence of ActRIIB, wherein the at least one N-X-S/T sequence is at a position outside of the ligand-binding pocket, wherein the protein comprises a N at the position corresponding to position 24 of SEQ ID NO;

2 and a S or T at the position corresponding to position 26 of SEQ ID NO;

2 and wherein the variant ActRIIB protein inhibits signaling by myostatin and/or GDF11 in a cell-based assay.

2. The variant ActRIIB protein of claim 1, wherein the protein comprises a R at the position corresponding to position 64 of SEQ ID NO:

2.

3. The variant ActRIIB protein of claim 1, wherein the protein comprises at least one additional alteration with respect to the sequence of SEQ ID NO:

2, and wherein the at least one additional alteration is a conservative alteration positioned within the ligand binding pocket.

4. The variant ActRIIB protein of claim 1, wherein the protein comprises at least one additional alteration with respect to the sequence of SEQ ID NO:

2, and wherein the at least one additional alteration is an alteration at one or more positions selected from an amino acid corresponding to K74, R40, Q53, K55, F82 and L79 of SEQ ID NO;

2.

5. The variant ActRIIB protein of claim 1, wherein the protein comprises an amino acid sequence beginning at an amino acid corresponding to any of amino acids 22-25 of SEQ ID NO:

2 and ending at an amino acid corresponding to any of amino acids 131, 133, or 134 of SEQ ID NO;

2.

6. The variant ActRIIB protein of claim 1, wherein the protein is a fusion protein further comprising a heterologous portion.

7. The variant ActRIIB protein of claim 6, wherein the protein is a homodimer.

8. The variant ActRIIB protein of claim 6, wherein the heterologous portion comprises a constant region from an IgG heavy chain.

9. The variant ActRIIB protein of claim 1, wherein the protein comprises an amino acid sequence that is at least 98% identical to the sequence of amino acids 29-109 of SEQ ID NO:

2.

10. The variant ActRIIB protein of claim 1, wherein the protein comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:

5.

11. The variant ActRIIB protein of claim 10, wherein the protein has an N at the position corresponding to position 5 of SEQ ID NO:

5.

12. The variant ActRIIB protein of claim 10, wherein the protein has a D the position corresponding to position 60 of SEQ ID NO:

5.

13. The variant ActRIIB protein of claim 10, wherein the protein has an E at the position corresponding to position 60 of SEQ ID NO:

5.

14. The variant ActRIIB protein of claim 10, wherein the protein is a dimer.

15. The variant ActRIIB protein of claim 11, wherein the protein is a dimer.

16. The variant ActRIIB protein of claim 12, wherein the protein is a dimer.

17. The variant ActRIIB protein of claim 13, wherein the protein is a dimer.

18. A pharmaceutical preparation comprising the protein of claim 1.

19. The variant ActRIIB protein of claim 1, wherein the protein comprises a K at the position corresponding to position 64 of SEQ ID NO:

2.

20. The variant ActRIIB protein of claim 1, wherein the protein comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:

5.

21. The variant ActRIIB protein of claim 1, wherein the protein comprises an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO:

5.

22. The variant ActRIIB protein of claim 1, wherein the protein comprises an amino acid sequence that is at least 95% identical to the amino acid sequence of SEQ ID NO:

12.

23. The variant ActRIIB protein of claim 1, wherein the protein comprises an amino acid sequence that is at least 99% identical to the amino acid sequence of SEQ ID NO:

12.

24. The variant ActRIIB protein of claim 1, wherein the protein comprises one or more modified amino acid residues selected from:

a glycosylated amino acid, a PEGylated amino acid, a farnesylated amino acid, an acetylated amino acid, a biotinylated amino acid, and an amino acid conjugated to a lipid moiety.

25. The variant ActRIIB protein of claim 6, wherein the heterologous portion comprises an Fc domain.

26. The variant ActRIIB protein of claim 25, wherein the Fc domain comprises the amino acid sequence of SEQ ID NO:

13.

27. The variant ActRIIB protein of claim 6, wherein the fusion protein further comprises a linker domain positioned between the ActRIIB polypeptide and the heterologous polypeptide.

28. The variant ActRIIB protein of claim 1, wherein the protein inhibits signaling by myostatin in a cell-based assay.

29. The variant ActRIIB protein of claim 1, wherein the protein inhibits signaling by GDF11 in a cell-based assay.