Fibroblast growth factor 21 mutations

Patent metadata

Field	Value
Publication number	US-9023791-B2
Application number	US-201113296343-A
Country	US
Kind code	B2
Filing date	Nov 15, 2011
Priority date	Nov 19, 2010
Publication date	May 5, 2015
Grant date	May 5, 2015

Abstract

The present invention provides novel polypeptide and protein variants of fibroblast growth factor 21 (FGF21) and pharmaceutical compositions comprising FGF21 polypeptide and protein variants.

First claim

What is claimed is:

1. A polypeptide variant having an amino acid sequence comprising SEQ ID NO:39.

2. The variant of claim 1 , wherein the variant further comprises one or more of the following modifications: (a) an amino-terminal truncation of no more than 5 amino acid residues; and (b) a carboxyl-terminal truncation of no more than 12 amino acid residues.

3. The variant of claim 1 , wherein the variant is covalently linked to polyethylene glycol (PEG) or polysialic acid.

4. The variant of claim 3 , wherein the polyethylene glycol (PEG) or polysialic acid comprises a branched, 40 kDa polyethylene glycol (PEG) or polysialic acid, and wherein said polyethylene glycol (PEG) or polysialic acid is covalently linked to a cysteine of the variant.

5. The variant of claim 3 , wherein the PEG is attached to the cysteine residue at position 122 of SEQ ID NO:39.

6. The variant of claim 5 , wherein the variant further comprises a branched, 40 kDa PEG group at position 122 of SEQ ID NO:39.

7. The variant of claim 1 , wherein the variant is fused to a heterologous protein consisting of one of the following: an IgG constant domain or fragment thereof; Human Serum Albumin (HSA); and an albumin-binding polypeptides.

8. The variant of claim 7 , wherein the heterologous protein is fused to the amino terminus of the variant.

9. The variant of claim 7 , wherein the heterologous protein is fused to the carboxy terminus of the variant.

10. A multimer consisting of the variant of claim 1 .

11. The multimer of claim 10 , wherein the multimer is a homodimer.

12. A pharmaceutical composition comprising the variant of SEQ ID NO:39.

13. The pharmaceutical composition of claim 12 , wherein the variant has a PEG group attached at the cysteine residue at position 122 of SEQ ID NO:39.

14. A polypeptide variant having an amino acid sequence selected from the group consisting of SEQ ID NO:16-19, 38, or 39.

15. The variant of claim 14 , wherein the variant further comprises one or more of the following modifications: (a) an amino-terminal truncation of no more than 5 amino acid residues; and (b) a carboxyl-terminal truncation of no more than 12 amino acid residues.

16. The variant of claim 14 , wherein the variant is covalently linked to polyethylene glycol (PEG) or polysialic acid.

17. The variant of claim 16 , wherein the polyethylene glycol (PEG) or polysialic acid comprises a branched, 40 kDa polyethylene glycol (PEG) or polysialic acid, and wherein said polyethylene glycol (PEG) or polysialic acid is covalently linked to a cysteine of the variant.

18. The variant of claim 16 , wherein the PEG is attached to the cysteine residue at position 122 of SEQ ID NO:39.

19. The variant of claim 18 , wherein the variant further comprises a branched, 40 kDa PEG group at position 122 of SEQ ID NO:39.

20. The variant of claim 14 , wherein the variant is fused to a heterologous protein consisting of one of the following: an IgG constant domain or fragment thereof; Human Serum Albumin (HSA); and an albumin-binding polypeptide.

21. The variant of claim 20 , wherein the heterologous protein is fused to the amino terminus of the variant.

22. The variant of claim 20 , wherein the heterologous protein is fused to the carboxy terminus of the variant.

23. A multimer consisting of at least one of the variants of claim 14 .

24. The multimer of claim 23 , wherein the multimer is a homodimer.

25. A pharmaceutical composition comprising a variant having a sequence selected from the group consisting of SEQ ID NO:16-19, 38, and 39.

26. The pharmaceutical composition of claim 25 , wherein the variant has a PEG group attached at the cysteine residue at position 122 of SEQ ID NO:39.

27. A polypeptide variant comprising a 4 amino acid N-terminally truncated mature FGF21 wild-type protein with the following substitutions made relative to SEQ ID NO:1: a glutamic acid residue substitution at position 56, a histidine residue substitution at position 74, a lysine residue substitution at position 105, an arginine residue substitution at position 150, a glutamine residue substitution at position 159, an alanine residue substitution at position 195, and one or more amino acid substitution that is:
(a) a glutamic acid residue substitution at position 82;
(b) an alanine residue substitution at position 98;
(c) an asparagine residue substitution or a glutamic acid substitution at position 130;
(d) a cysteine residue substitution at position 154; or
(e) a proline residue substitution at position 174.

28. The variant of claim 27 , wherein the variant further comprises one or more of the following modifications: (a) an amino-terminal truncation of no more than 5 amino acid residues; and (b) a carboxyl-terminal truncation of no more than 12 amino acid residues.

29. The variant of claim 27 , wherein the variant is covalently linked to polyethylene glycol (PEG) or polysialic acid.

30. The variant of claim 29 , wherein the polyethylene glycol (PEG) or polysialic acid comprises a branched, 40 kDa polyethylene glycol (PEG) or polysialic acid, and wherein said polyethylene glycol (PEG) or polysialic acid is covalently linked to a cysteine of the variant.

31. The variant of claim 27 , wherein the variant is fused to a heterologous protein consisting of one of the following: an IgG constant domain or fragment thereof; Human Serum Albumin (HSA); and an albumin-binding polypeptides.

32. The variant of claim 31 , wherein the heterologous protein is fused to the amino terminus of the variant.

33. The variant of claim 31 , wherein the heterologous protein is fused to the carboxy terminus of the variant.

34. A multimer consisting of at least one of the variants of claim 27 .

35. The multimer of claim 34 , wherein the multimer is a homodimer.

36. A variant of claim 27 having the cysteine residue substitution at position 122 of SEQ ID NO:39, wherein the variant is covalently linked to polyethylene glycol (PEG) or polysialic acid attached to said cysteine residue.

37. The variant of claim 36 , wherein the variant further comprises a branched, 40 kDa PEG group at position 122 of SEQ ID NO:39.

38. A pharmaceutical composition comprising the variant of claim 27 .

39. A pharmaceutical composition comprising the variant of claim 27 having the cysteine residue substitution at position 122 of SEQ ID NO:39, wherein the variant is covalently linked to polyethylene glycol (PEG) or polysialic acid attached to said cysteine residue.

Assignees

• Boettcher Brian R
• Caplan Shari L
• Daniels Douglas S
• Geierstanger Bernhard H
• Hamamatsu Norio
• Licht Stuart
• Loew Andreas
• Weldon Stephen Craig
• Novartis Ag
• Irm Llc

Inventors

• Boettcher Brian R
• Caplan Shari L
• Daniels Douglas S
• Geierstanger Bernhard H
• Hamamatsu Norio
• Licht Stuart
• Loew Andreas
• Weldon Stephen Craig

Classifications

• A61P5/50

  for increasing or potentiating the activity of insulin · CPC title

• A61P3/06

  Antihyperlipidemics · CPC title

• A61P3/10

  for hyperglycaemia, e.g. antidiabetics · CPC title

• A61P3/04

  Anorexiants; Antiobesity agents · CPC title

• A61P3/00

  Drugs for disorders of the metabolism (of the blood or the extracellular fluid A61P7/00) · CPC title