Mutated fibroblast growth factor (fgf) 1 and methods of use

Abstract

The present disclosure provides FGF1 mutant proteins, such as those having an N-terminal deletion, point mutation(s), or combinations thereof, which can reduce blood glucose in a mammal. Such mutant FGF1 proteins can be part of a chimeric protein that includes a β-Klotho-binding protein, an FGFR1c-binding protein, a β-Klotho-binding protein and a FGFR1c-binding protein, a C-terminal region from FGF19 or FGF21. In some examples, mutant FGF1 proteins have reduced mitogenic activity. Also provided are nucleic acid molecules that encode such proteins, and vectors and cells that include such nucleic acids. Methods of using the disclosed molecules to reduce blood glucose levels are also provided.

We claim: 
     
         1 . An isolated mutated mature fibroblast growth factor (FGF) 1 protein comprising:
 a deletion of at least six consecutive N-terminal amino acids;   a point mutation at R35; and   a point mutation at K112, K113, K118, R119 or combinations thereof, wherein the amino acid numbering is based on SEQ ID NO: 5   
     
     
         2 . The isolated mutated mature FGF1 protein of  claim 1 , wherein the a deletion of at least six contiguous N-terminal amino acids comprises deletion of at least 9 consecutive N-terminal amino acids. 
     
     
         3 . The isolated mutated mature FGF1 protein of  claim 1 , wherein the a deletion of at least six contiguous N-terminal amino acids comprises deletion of 9 consecutive N-terminal amino acids. 
     
     
         4 . The isolated mutated mature FGF1 protein of  claim 1 , wherein the point mutation at R35 is R35E. 
     
     
         5 . The isolated mutated mature FGF1 protein of  claim 1 , wherein the point mutation at K112 is K112D, K112E, or K112Q. 
     
     
         6 . The isolated mutated mature FGF1 protein of  claim 1 , wherein the point mutation at K113 is K113D, K113E, or K113Q. 
     
     
         7 . The isolated mutated mature FGF1 protein of  claim 1 , wherein the point mutation at R119 is R119G, R119V, or R119E. 
     
     
         8 . The isolated mutated mature FGF1 protein of  claim 1 , wherein the isolated protein comprises a point mutation at both K112 and K113. 
     
     
         9 . The isolated mutated mature FGF1 protein of  claim 1 , wherein the isolated protein comprises a point mutation at:
 K113;   K118;   R119;   K113 and R119; or   K118 and R119,   
     
     
         10 . The isolated mutated mature FGF1 protein of  claim 8 , wherein the point mutation at K112 is K112Q and the point mutation at K113 is K113Q. 
     
     
         11 . The isolated mutated mature FGF1 protein of  claim 1 , wherein the protein comprises at least 90% sequence identity to SEQ ID NO: 194. 
     
     
         12 . The isolated mutated mature FGF1 protein of  claim 1 , wherein the protein comprises at least 95% sequence identity to SEQ ID NO: 194. 
     
     
         13 . The isolated mutated mature FGF1 protein of  claim 1 , wherein the protein comprises at least 98% sequence identity to SEQ ID NO: 194. 
     
     
         14 . A method of reducing blood glucose in a mammal, comprising:
 administering to the subject a therapeutically effective amount of the isolated mutated mature FGF1 protein of  claim 1 , thereby reducing blood glucose in the mammal.   
     
     
         15 . The method of  claim 14 , wherein the administering is subcutaneous, intraperitoneal, intramuscular, or intravenous. 
     
     
         16 . The method of  claim 14 , wherein the amount of FGF1 protein administered is at least 0.5 mg/kg. 
     
     
         17 . The method of  claim 14 , wherein the method further comprises administering an additional therapeutic compound. 
     
     
         18 . The method of  claim 17 , wherein the additional therapeutic compound is an alpha-glucosidase inhibitor, amylin agonist, dipeptidyl-peptidase 4 (DPP-4) inhibitor, meglitinide, sulfonylurea, or a peroxisome proliferator-activated receptor (PPAR)-gamma agonist. 
     
     
         19 . The method of  claim 18 , wherein the PPAR-gamma agonist is a thiazolidinedione (TZD), aleglitazar, farglitazar, muraglitazar, or tesaglitazar. 
     
     
         20 . The method of  claim 19 , wherein the TZD is pioglitazone, rosiglitazone, rivoglitazone, or troglitazone. 
     
     
         21 . The method of  claim 14 , wherein the mammal is a human.