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