Fgf21 mutants and uses thereof

Patent metadata

Field	Value
Publication number	US-2016244497-A1
Application number	US-201615007438-A
Country	US
Kind code	A1
Filing date	Jan 27, 2016
Priority date	Oct 10, 2008
Publication date	Aug 25, 2016
Grant date	—

Abstract

The invention provides nucleic acid molecules encoding FGF21 mutant polypeptides, FGF21 mutant polypeptides, pharmaceutical compositions comprising FGF21 mutant polypeptides, and methods for treating metabolic disorders using such nucleic acids, polypeptides, or pharmaceutical compositions.

First claim

What is claimed is:

1 . An isolated nucleic acid molecule comprising a nucleotide sequence encoding a polypeptide of SEQ ID NO: 4 having at least one amino acid substitution that is:
(a) a lysine residue at one or more of positions 36, 72, 77, 126 and 175; (b) a cysteine residue at one or more of positions 37, 38, 46, 91, 69, 77, 79, 87, 91, 112, 113, 120, 121, 125, 126, 175, 170, and 179; (c) an arginine residue at one or more of positions 56, 59, 69, and 122; (d) a glycine residue at position 170; (e) a glycine residue at position 171;
and combinations of (a)-(e).

2 . A vector comprising the nucleic acid molecule of claim 1 .

3 . A host cell comprising the vector of claim 2 .

4 . The host cell of claim 3 that is a eukaryotic cell.

5 . The host cell of claim 3 that is a prokaryotic cell.

6 . A process of producing a polypeptide encoded by the vector of claim 2 comprising culturing a host cell comprising the vector of claim 2 under suitable conditions to express the polypeptide, and optionally isolating the polypeptide.

7 . A polypeptide produced by the process of claim 6 .

8 . The polypeptide of claim 7 , further comprising a proline or glycine residue added to the C-terminus of the polypeptide.

9 . An isolated polypeptide comprising the amino acid sequence of SEQ ID NO: 4 having at least one amino acid substitution that is:
(a) a lysine residue at one or more of positions 36, 72, 77, 126 and 175; (b) a cysteine residue at one or more of positions 37, 38, 46, 91, 69, 77, 79, 87, 91, 112, 113, 120, 121, 125, 126, 175, 170, and 179; (c) an arginine residue at one or more of positions 56, 59, 69, and 122; (d) a glycine residue at position 170; (e) a glycine residue at position 171;
and combinations of (a)-(e),

10 . The polypeptide of claim 9 , further comprising a proline or glycine residue added to the C-terminus of the polypeptide.

11 . The isolated polypeptide of claim 9 , wherein the polypeptide is covalently linked to one or more polymers.

12 . The isolated polypeptide of claim 11 , wherein the polypeptide is covalently linked to one polymer.

13 . The isolated polypeptide of claim 12 , wherein the polymer is a water-soluble polymer.

14 . The isolated polypeptide of claim 13 , wherein the water-soluble polymer is polyethylene glycol (PEG), monomethoxy-polyethylene glycol, dextran, cellulose, poly-(N-vinyl pyrrolidone) polyethylene glycol, propylene glycol homopolymers, polypropylene oxide/ethylene oxide co-polymers, polyoxyethylated polyols, or polyvinyl alcohol.

15 . The isolated polypeptide of claim 14 , wherein the water-soluble polymer is PEG.

16 . The isolated polypeptide of claim 11 , wherein the polymer is a branched polymer.

17 . The isolated polypeptide of claim 9 , wherein the polypeptide has a PEG moiety covalently linked to its amino-terminus.

18 . The isolated polypeptide of claim 9 , wherein the polypeptide is covalently linked to two polymers.

19 . The isolated polypeptide of claim 18 , wherein one if the two polymers is a water-soluble polymer.

20 . The isolated polypeptide of claim 19 , wherein the water-soluble polymer is polyethylene glycol (PEG), monomethoxy-polyethylene glycol, dextran, cellulose, poly-(N-vinyl pyrrolidone) polyethylene glycol, propylene glycol homopolymers, polypropylene oxide/ethylene oxide co-polymers, polyoxyethylated polyols, or polyvinyl alcohol.

21 . The isolated polypeptide of claim 20 , wherein the water-soluble polymer is PEG.

22 . The isolated polypeptide of claim 18 , wherein one of the polymers is branched.

23 . The isolated polypeptide of claim 18 , wherein both of the polymers are branched.

24 . The isolated polypeptide of claim 9 , wherein the polypeptide has a PEG moiety covalently linked to its amino-terminus.

25 . A pharmaceutical composition comprising the isolated polypeptide of claim 9 and a pharmaceutically acceptable formulation agent.

26 . The pharmaceutical composition of claim 25 , wherein the pharmaceutically acceptable formulation agent is a carrier, adjuvant, solubilizer, stabilizer, or anti-oxidant.

27 . A method for treating a metabolic disorder comprising administering to a human patient in need thereof the pharmaceutical composition of claim 26 .

28 . The method of claim 27 , wherein the metabolic disorder is diabetes.

29 . The method of claim 27 , wherein the metabolic disorder is obesity.

30 . An isolated nucleic acid encoding a polypeptide comprising the amino acid sequence of SEQ ID NO: 4 having at least one amino acid substitution that is:
(a) a lysine residue at one or more of positions 36, 72, 77, 126 and 175; (b) a cysteine residue at one or more of positions 37, 38, 46, 91, 69, 77, 79, 87, 91, 112, 113, 120, 121, 125, 126, 175, 170, and 179; (c) an arginine residue at one or more of positions 56, 59, 69, and 122; (d) a glycine residue at position 170; (e) a glycine residue at position 171;
and combinations of (a)-(e),
and which comprises additions, deletions or further substitutions that make the polypeptide at least 85% identical to SEQ ID NO:4, provided that the at least one amino acid substitution of claim 1 (a)-(e) is not further modified.

31 . A vector comprising the nucleic acid molecule of claim 30 .

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

33 . The host cell of claim 32 that is a eukaryotic cell.

34 . The host cell of claim 32 that is a prokaryotic cell.

35 . A process of producing a polypeptide encoded by the vector of claim 30 comprising culturing a host cell comprising the vector of claim 30 under suitable conditions to express the polypeptide, and optionally isolating the polypeptide.

36 . A polypeptide produced by the process of claim 35 .

37 . The polypeptide of claim 36 , further comprising a proline or glycine residue added to the C-terminus of the polypeptide.

38 . An isolated polypeptide comprising the amino acid sequence of SEQ ID NO: 4 having at least one amino acid substitution that is:
(a) a lysine residue at one or more of positions 36, 72, 77, 126 and 175; (b) a cysteine residue at one or more of positions 37, 38, 46, 91, 69, 77, 79, 87, 91, 112, 113, 120, 121, 125, 126, 175, 170, and 179; (c) an arginine residue at one or more of positions 56, 59, 69, and 122; (d) a glycine residue at position 170; (e) a glycine residue at position 171;
and combinations of (a)-(e),
and which comprises additions, deletions or further substitutions that make the polypeptide at least 85% identical to SEQ ID NO:4, provided that the at least one amino acid substitution of claim 1 (a)-(e) is not further modified.

39 . The polypeptide of claim 38 , further comprising a proline or glycine residue added to the C-terminus of the polypeptide.

40 . The isolated polypeptide of claim 38 , wherein the polypeptide is covalently linked to one polymer.

41 . The isolated polypeptide of claim 40 , wherein the polymer is a water-soluble polymer.

42 . The isolated polypeptide of claim 41 , wherein the water-soluble polymer is polyethylene glycol (PEG), monomet

Assignees

• Amgen Inc

Inventors

• Walker Kenneth W
• Gegg Jr Colin V
• Hecht Randy I
• Belouski Edward J
• Li Yue-Sheng
• Michaels Mark L
• Xu Jing
• Ellison Murielle Marie

Classifications

• A61P3/10

  for hyperglycaemia, e.g. antidiabetics · CPC title

• A61P3/00

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

• A61P3/04

  Anorexiants; Antiobesity agents · CPC title

• A61K38/00

  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 title

• C07K14/50 Primary

  Fibroblast growth factor [FGF] · CPC title