Fibroblast growth factor 1 (fgf1) mutant proteins that selectively activate fgfr1b to reduce blood glucose

Abstract

The present disclosure provides FGF1 mutant proteins, which selectively bind to/activate FGFR1b. Also provided are nucleic acid molecules that encode such proteins, and vectors and cells that include such nucleic acids. Methods of using the disclosed FGF1 mutants to reduce blood glucose in a mammal and treat a metabolic disorder are provided.

We claim: 
     
         1 . An isolated mutated mature fibroblast growth factor (FGF) 1 protein, comprising:
 at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 99%, or 100% sequence identity to SEQ ID NO: 10, and retaining the N7H, N18D, D32E, Y64C, F85L, E90G, K101R, and N114S amino acid substitutions;   at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 99%, or 100% sequence identity to SEQ ID NO: 11, and retaining the K10E, L13P, L44P, I98T, and K113Q amino acid substitutions;   at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 99%, or 100% sequence identity to SEQ ID NO: 12, and retaining the N1D, G19R, and K118E amino acid substitutions; or   at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 99%, or 100% sequence identity to SEQ ID NO: 13, and retaining the S76P amino acid substitution.   
     
     
         2 . The isolated protein of  claim 1 , wherein the protein is 90 to 140 amino acids in length. 
     
     
         3 . The isolated protein of  claim 1 , wherein the mutated mature FGF1 protein comprises a deletion of 6, 7, 8, 9, 10, 11, or 12 contiguous N-terminal amino acids from a native FGF1 protein. 
     
     
         4 . The isolated protein of  claim 1 , wherein the mutated mature FGF1 protein further comprises at least one additional amino acid substitution selected from N7H, K10E, L13P, N18D, G19R, D32E, L44P, Y64C, S76P, F85L, E90G, I98T, K101R, K113Q, N114S, and K118E. 
     
     
         5 . The isolated protein of  claim 3 , wherein the deleted contiguous N-terminal amino acids are replaced with the amino acid sequence comprising MRDSSPL (SEQ ID NO: 14), SYNHLQGDVR (SEQ ID NO: 15), SYNHLQGDVRV (SEQ ID NO: 16), or SYDYMEGGDIRV (SEQ ID NO: 17). 
     
     
         6 . The isolated protein of  claim 3 , wherein the deleted contiguous N-terminal amino acids are replaced with the amino acid sequence consisting of MRDSSPL (SEQ ID NO: 14), SYNHLQGDVR (SEQ ID NO: 15), SYNHLQGDVRV (SEQ ID NO: 16), or SYDYMEGGDIRV (SEQ ID NO: 17). 
     
     
         7 . The isolated protein of  claim 1 , wherein the protein has
 decreased mitogenicity compared to a native mature FGF1 protein (SEQ ID NO: 2, 4, 5, 6, 7, 8 or 9); and   increased blood glucose lowering ability compared to a native mature FGF1 protein (SEQ ID NO: 2, 4, 5, 6, 7, 8 or 9).   
     
     
         8 . The isolated protein of  claim 1 , wherein the protein has decreased mitogenicity compared to a native mature FGF1 protein (SEQ ID NO: 2, 4, 5, 6, 7, 8 or 9). 
     
     
         9 . The isolated protein of  claim 1 , wherein the protein has increased blood glucose lowering ability compared to a native mature FGF1 protein (SEQ ID NO: 2, 4, 5, 6, 7, 8 or 9) both. 
     
     
         10 . The isolated protein of  claim 1 , wherein the protein comprises at least 95% sequence identity to SEQ ID NO: 10, and retains the N7H, N18D, D32E, Y64C, F85L, E90G, K101R, and N114S amino acid substitutions. 
     
     
         11 . The isolated protein of  claim 1 , wherein the protein comprises at least 95% sequence identity to SEQ ID NO: 11, and retains the K10E, L13P, L44P, I98T, and K113Q amino acid substitutions. 
     
     
         12 . The isolated protein of  claim 1 , wherein the protein comprises at least 95% sequence identity to SEQ ID NO: 12, and retains the N1D, G19R, and K118E amino acid substitutions. 
     
     
         13 . The isolated protein of  claim 1 , wherein the protein comprises at least 95% sequence identity to SEQ ID NO: 13, and retains the S76P amino acid substitution.