Fgf mutants and uses thereof

Patent metadata

Field	Value
Publication number	US-2016168223-A1
Application number	US-201615004281-A
Country	US
Kind code	A1
Filing date	Jan 22, 2016
Priority date	Jun 4, 2008
Publication date	Jun 16, 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 polypeptide comprising an amino acid sequence of SEQ ID NO:4, further comprising the substitution of any amino acid for: the alanine residue at position 45, the leucine residue at position 86, the leucine residue at position 98, the alanine residue at position 111, the alanine residue at position 129, the glycine residue at position 170, the proline residue at position 171 or the serine residue at position 172, and combinations thereof.

2 . The isolated polypeptide of claim 1 , wherein the isolated polypeptide comprises the substitution of any amino acid for: the leucine residue at position 98, the proline residue at 171 or both the leucine residue at position 98 and the proline residue at position 171.

3 . The isolated polypeptide of claim 2 , wherein the isolated polypeptide comprises the substitution of any amino acid for both the leucine residue at position 98 and the proline residue at position 171.

4 . An isolated polypeptide comprising an amino acid sequence of SEQ ID NO: 4 having:
(a) at least one amino acid substitution that is:
(i) a glutamine, isoleucine, or lysine residue at position 19;
(ii) a histidine, leucine, or phenylalanine residue at position 20;
(iii) an isoleucine, phenylalanine, tyrosine, or valine residue at position 21;
(iv) an isoleucine, phenylalanine, or valine residue at position 22;
(v) an alanine or arginine residue at position 150;
(vi) an alanine or valine residue at position 151;
(vii) a histidine, leucine, phenylalanine, or valine residue at position 152;
(viii) an alanine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, proline, or serine residue at position 170;
(ix) an alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, lysine, serine, threonine, tryptophan, or tyrosine residue at position 171;
(x) a leucine or threonine residue at position 172; or
(xi) an arginine or glutamic acid residue at position 173; and
(b) at least one amino acid substitution that is:
(i) an arginine, glutamic acid, or lysine residue at position 26;
(ii) an arginine, glutamic acid, glutamine, lysine, or threonine residue at position 45;
(iii) a threonine residue at position 52;
(iv) a cysteine, glutamic acid, glycine, or serine residue at position 58;
(v) an alanine, arginine, glutamic acid, or lysine residue at position 60;
(vi) an alanine, arginine, cysteine, or histidine residue at position 78;
(vii) a cysteine or threonine residue at position 86;
(viii) an alanine, arginine, glutamic acid, lysine, or serine residue at position 88;
(ix) an arginine, cysteine, glutamic acid, glutamine, lysine, or threonine residue at position 98;
(x) an arginine, aspartic acid, cysteine, or glutamic acid residue at position 99;
(xi) a lysine or threonine residue at position 111;
(xii) an arginine, asparagine, aspartic acid, glutamic acid, glutamine, histidine, or lysine residue at position 129; or
(xiii) an arginine, glutamic acid, histidine, lysine, or tyrosine residue at position 134;
and combinations thereof.

5 . The isolated polypeptide of claim 4 , wherein the residue at position 98 is arginine and the residue at position 171 is proline.

6 . The isolated polypeptide of claim 4 , wherein the polypeptide comprises an amino acid sequence that is at least 85 percent identical to the amino acid sequence of SEQ ID NO: 4, but wherein the at least one amino acid substitution of claim 4 (a)(i)-(xi) and 4 (b)(i)-(xiii) is not further modified.

7 . An isolated polypeptide comprising an amino acid sequence of SEQ ID NO: 4 having at least one amino acid substitution that is:
(a) a glutamine, lysine or isoleucine residue at position 19; (b) a histidine, leucine, or phenylalanine residue at position 20; (c) an isoleucine, phenylalanine, tyrosine, or valine residue at position 21; (d) an isoleucine, phenylalanine, or valine residue at position 22; (e) an alanine or arginine residue at position 150; (f) an alanine or valine residue at position 151; (g) a histidine, leucine, phenylalanine, or valine residue at position 152; (h) an alanine, aspartic acid, cysteine, or proline residue at position 170; (i) an alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, lysine, serine, threonine, tryptophan, or tyrosine residue at position 171; (j) a leucine residue at position 172; or (k) an arginine or glutamic acid residue at position 173; and combinations thereof.

8 . The isolated polypeptide of claim 7 , wherein the residue at position 171 is proline.

9 . The isolated polypeptide of claim 7 , wherein the polypeptide comprises an amino acid sequence that is at least 85 percent identical to the amino acid sequence of SEQ ID NO: 4, but wherein the at least one amino acid substitution of claim 7 (a)-(k) is not further modified.

10 . An isolated polypeptide comprising an amino acid sequence of SEQ ID NO: 4 having at least one amino acid substitution that is:
(a) an arginine, glutamic acid, or lysine residue at position 26; (b) an arginine, glutamic acid, glutamine, lysine, or threonine residue at position 45; (c) a threonine residue at position 52; (d) a glutamic acid, glycine, or serine residue at position 58; (e) an alanine, arginine, glutamic acid, or lysine residue at position 60; (f) an alanine, arginine, or histidine residue at position 78; (g) an alanine residue at position 88; (h) an arginine, glutamic acid, glutamine, lysine, or threonine residue at position 98; (i) an arginine, aspartic acid, cysteine, or glutamic acid residue at position 99; (j) a lysine or threonine residue at position 111; (k) an arginine, asparagine, aspartic acid, glutamic acid, glutamine, histidine, or lysine residue at position 129; or (l) an arginine, glutamic acid, histidine, lysine, or tyrosine residue at position 134; and combinations thereof.

11 . The isolated polypeptide of claim 10 , wherein the residue at position 98 is arginine.

12 . The isolated polypeptide of claim 10 , wherein the polypeptide comprises an amino acid sequence that is at least 85 percent identical to the amino acid sequence of SEQ ID NO: 4, but wherein the at least one amino acid substitution of claim 10 (a)-(l) is not further modified.

13 . The isolated polypeptide of any of claims 1 , 4 , 7 or 10 , further comprising at least one amino acid substitution that is:
(a) a phenylalanine, proline, alanine, serine or glycine at position 179;
(b) a glutamic acid, glycine, proline, or serine at position 180; or
(c) a lysine, glycine, threonine, alanine, leucine, or proline at position 181.

14 . The isolated polypeptide of any of claims 1 , 4 , 7 or 10 , further comprising 1 to 10 amino acid residues fused to the C-terminus of the polypeptide.

15 . The isolated polypeptide of claim 10 , wherein the 1 to 10 amino acid residues are selected from the group consisting of glycine, proline and combinations thereof.

16 . The isolated polypeptide of any of claims 1 , 4 , 7 or 10 , wherein the polypeptide comprises:
(a) an amino-terminal truncation of no more than 8 amino acid residues, wherein the polypeptide is capable of lowering blood glucose in a mammal;
(b) a carboxyl-terminal truncation of no more than 12 amino acid residues, wherein the polypeptide is capable of lowering blood glucose in a mammal; or
(c) an amino-terminal truncation of no more than 8 amino acid residues and a carboxyl-terminal truncation of no more than 12 amino acid residues, wherein the polypeptide is capable of lowering bl

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

• C07K16/18

  against material from animals or humans · CPC title

• A61P3/10

  for hyperglycaemia, e.g. antidiabetics · CPC title

• 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

• 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