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