FGF mutants with reduced proteolysis and aggregation

Patent metadata

Field	Value
Publication number	US-9273106-B2
Application number	US-201314134482-A
Country	US
Kind code	B2
Filing date	Dec 19, 2013
Priority date	Jun 4, 2008
Publication date	Mar 1, 2016
Grant date	Mar 1, 2016

Abstract

The invention provides FGF21 mutant polypeptides with reduced proteolysis and aggregation, pharmaceutical compositions comprising the same.

First claim

What is claimed is:

1. An isolated fusion protein comprising:
(a) an IgG constant domain;
(b) a linker sequence fused to the IgG constant domain; and
(c) an FGF21 mutant fused to the linker sequence and wherein the FGF21 mutant comprises the amino acid sequence of SEQ ID NO: 4, wherein an arginine residue has been substituted for the leucine residue at position 98, and wherein a glutamic acid residue has been substituted for the glycine at position 170.

2. The isolated fusion protein of claim 1 , further comprising
(i) an N-terminal truncation of 8 or fewer residues;
(ii) a C terminal truncation of 12 or fewer residues; or
(iii) an N-terminal truncation of 8 or fewer residues and a C terminal truncation of 12 or fewer residues.

3. The isolated fusion protein of claim 2 , wherein the polypeptide is capable of lowering blood glucose in a mammal.

4. The isolated fusion protein of claim 1 , wherein the polypeptide is covalently linked to one or more polymers.

5. The isolated fusion protein of claim 4 , wherein the polymer is PEG.

6. A pharmaceutical composition comprising the isolated fusion protein of claim 1 and a pharmaceutically acceptable formulation agent.

7. A method selected from the group consisting of
reducing triglyceride levels in a patient,
improving glucose tolerance in a patient,
lowering body weight in a patient, and
lowering insulin levels in a patient,
said method comprising administering to a human patient in need thereof the pharmaceutical composition of claim 6 .

8. The method of claim 7 , wherein said patient has diabetes.

9. The method of claim 7 , wherein said patient is obese.

10. The isolated fusion protein of claim 1 , further comprising a lysine residue substituted for the alanine at position 45.

11. The isolated fusion protein of claim 10 , further comprising
(i) an N-terminal truncation of 8 or fewer residues;
(ii) a C terminal truncation of 12 or fewer residues; or
(iii) an N-terminal truncation of 8 or fewer residues and a C terminal truncation of 12 or fewer residues.

12. The isolated fusion protein of claim 11 , wherein the polypeptide is capable of lowering blood glucose in a mammal.

13. The isolated fusion protein of claim 10 , wherein the polypeptide is covalently linked to one or more polymers.

14. The isolated fusion protein of claim 13 , wherein the polymer is PEG.

15. A pharmaceutical composition comprising the isolated fusion protein of claim 10 and a pharmaceutically acceptable formulation agent.

16. A method selected from the group consisting of
reducing triglyceride levels in a patient,
improving glucose tolerance in a patient,
lowering body weight in a patient, and
lowering insulin levels in a patient,
said method comprising administering to a human patient in need thereof the pharmaceutical composition of claim 15 .

17. The method of claim 16 , wherein said patient has diabetes.

18. The method of claim 16 , wherein said patient is obese.

19. An isolated nucleic acid encoding a fusion protein comprising:
(a) an IgG constant domain;
(b) a linker sequence fused to the IgG constant domain; and
(c) an FGF21 mutant fused to the linker sequence and wherein the FGF21 mutant comprises the amino acid sequence of SEQ ID NO: 4, wherein an arginine residue has been substituted for the leucine residue at position 98, and wherein a glutamic acid residue has been substituted for the glycine at position 170.

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

21. A host cell comprising the nucleic acid molecule of claim 19 .

Assignees

• Amgen Inc

Inventors

• Belouski Edward John
• Ellison Murielle Marie
• Hamburger Agnes Eva
• Hecht Randy Ira
• Li Yue-Sheng
• Michaels Mark Leo
• Sun Jeonghoon
• Xu Jing

Classifications

• C07K19/00

  Hybrid peptides {, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes} · CPC title

• C12N5/10

  Cells modified by introduction of foreign genetic material · CPC title

• C07K16/18

  against material from animals or humans · CPC title

• A61K47/42

  Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein (oligopeptides having up to five amino acids {A61K47/183}; polyamino acids A61K47/34) · CPC title

• C07K14/50 Primary

  Fibroblast growth factor [FGF] · CPC title