Chimeric fibroblast growth factors with altered receptor specificity

Abstract

The present invention is directed to novel chimeric fibroblast growth factor (FGF) polypeptides, novel DNA encoding chimeric FGF polypeptides, and to the recombinant production of chimeric FGF polypeptides, and to methods, compositions and assays utilizing chimeric FGF polypeptides for the therapeutic treatment of metabolic-related disorders and other conditions, and for producing pharmaceutically active compositions including chimeric FGF polypeptides, the compositions having therapeutic and pharmacologic properties including those associated with the treatment of metabolic-related disorders and other conditions.

1 . A chimeric fibroblast growth factor 19 (FGF19) polypeptide, the sequence of the polypeptide comprising:
 a C-terminal portion that comprises a C-terminal portion of the hFGF19 polypeptide sequence; and   an N-terminal portion that comprises an N-terminal portion of the hFGF21 polypeptide sequence, wherein   the C-terminal portion of the hFGF19 polypeptide sequence is from about 45 to about 185 residues in length, the C-terminal portion of the hFGF19 polypeptide sequence having a first position and a final position,   the N-terminal portion of the hFGF21 polypeptide sequence is from about 7 to about 140 residues in length, the C-terminal portion of the hFGF21 polypeptide sequence having a first position and a final position,   wherein the chimeric hFGF19 polypeptide does not substantially activate FGFR4 in either a Klotho-beta independent or Klotho-beta dependent manner, and   wherein the chimeric FGF19 polypeptide activates FGFR1c in a Klotho-beta dependent manner.   
     
     
         2 . The chimeric FGF19 polypeptide of  claim 1 , wherein:
 the first position of the C-terminal portion of the hFGF19 polypeptide sequence corresponds to a position of SEQ ID NO:1 selected from the group consisting of: 25, 26, 27, 28, 30, 33, 35, 37, 40, 41, 42, 43, 44, 45, 52, 53, 54, 56, 57, 58, 59, 72, 73, 74, 79, 80, 81, 143, 144, 145 and 146; and   the final position of the C-terminal portion of the hFGF19 polypeptide sequence corresponds to about position 194 of SEQ ID NO:1.   
     
     
         3 . The chimeric FGF19 polypeptide of  claim 2 , wherein:
 the first position of the C-terminal portion of the hFGF19 polypeptide sequence corresponds to position 25 of SEQ ID NO:1, and   the final position of the C-terminal portion of the hFGF19 polypeptide sequence corresponds to about position 194 of SEQ ID NO:1.   
     
     
         4 . The chimeric FGF19 polypeptide of  claim 1 , wherein:
 the first position of the N-terminal portion of the hFGF21 polypeptide sequence corresponds to about position 1 of SEQ ID NO:2; and   the final position of the N-terminal portion of the hFGF21 polypeptide sequence corresponds to a position of SEQ ID NO:2 selected from the group consisting of: 20, 21, 22, 23, 25, 27, 29, 31, 34, 35, 36, 37, 38, 39, 46, 47, 48, 50, 51, 52, 53, 66, 67, 68, 73, 74, 75, 135, 136, 137 and 138.   
     
     
         5 . The chimeric FGF19 polypeptide of  claim 4 , wherein:
 the first position of the N-terminal portion of the hFGF21 polypeptide sequence corresponds to about position 1 of SEQ ID NO:2; and   the final position of the N-terminal portion of the hFGF21 polypeptide sequence corresponds to position 20 of SEQ ID NO:2.   
     
     
         6 - 16 . (canceled) 
     
     
         17 . The chimeric FGF19 polypeptide of  claim 1 , wherein the chimeric hFGF19 polypeptide is fused to a second polypeptide, the second polypeptide is selected from the group consisting of: the Fc portion of an immunoglobulin, an analog of the Fc portion of an immunoglobulin and one or more fragments of the Fc portion of an immunoglobulin. 
     
     
         18 . The chimeric FGF19 polypeptide of  claim 17 , wherein the immunoglobulin is selected from the group consisting of: IgG-1, IgG-2, IgG-3, IgG-4, IgA-1, IgA-2, IgE, IgD and IgM. 
     
     
         19 . The chimeric FGF19 polypeptide of  claim 17 , wherein the Fc portion is human or humanized. 
     
     
         20 . The chimeric FGF19 polypeptide of  claim 17 , wherein the C-terminus of the chimeric hFGF19 polypeptide is fused to the N-terminus of the second polypeptide. 
     
     
         21 . The chimeric FGF19 polypeptide of  claim 20 , wherein the C-terminus of the chimeric hFGF19 polypeptide is fused to the N-terminus of the second polypeptide via a linker, the linker is selected from the group consisting of: a [Gly]n linker, a [Gly3Ser]m linker and a [Gly4Ser]m linker, wherein n is an integer from 1-30 and m is an integer from 1-6. 
     
     
         22 - 23 . (canceled) 
     
     
         24 . The chimeric FGF19 polypeptide of  claim 1 , wherein the chimeric FGF19 polypeptide when administered to an individual does not reduce the level of phosphorylated STAT5 polypeptide in the individual. 
     
     
         25 . The chimeric FGF19 polypeptide of  claim 1 , wherein the chimeric FGF19 polypeptide when administered to an individual reduces the amount of phosphorylated STAT5 polypeptide in the individual but this amount of phosphorylated STAT5 polypeptide is greater than the amount of phosphorylated STAT5 polypeptide upon administration of native hFGF21 to the individual. 
     
     
         26 . The chimeric FGF19 polypeptide of  claim 1 , wherein the chimeric FGF19 polypeptide when administered to an individual reduces the amount of phosphorylated STAT5 polypeptide to an amount that is any of: from 100% to 5%, from 100% to 10%, from 100% to 20%, from 100% to 30%, from 100% to 40%, from 100% to 50%, from 100% to 60%, from 100% to 70%, from 100% to 80%, from 100% to 90% or from 100% to 95%, of the amount of phosphorylated STAT5 polypeptide in the individual without such administration. 
     
     
         27 . The chimeric FGF19 polypeptide of  claim 1 , wherein the chimeric FGF19 polypeptide when administered to an individual, the reduction in the amount of phosphorylated STAT5 polypeptide is less than reduction in the amount of phosphorylated STAT5 polypeptide upon administration of native hFGF21. 
     
     
         28 . The chimeric FGF19 polypeptide of  claim 27 , wherein the reduction of the phosphorylated STAT5 polypeptide when the chimeric hFGF19 polypeptide is administered to the individual is by any of: from 0% to 5%, from 0% to 10%, from 0% to 20%, from 0% to 30%, from 0% to 40%, from 0% to 50%, from 0% to 60%, from 0% to 70%, from 0% to 80%, from 0% to 90% or from 0% to 95%, of the reduction in the amount of phosphorylated STAT5 polypeptide upon administration of native hFGF21. 
     
     
         29 . The chimeric FGF19 polypeptide of  claim 1 , wherein the chimeric FGF19 polypeptide when administered to an individual does not induce growth hormone resistance. 
     
     
         30 . The chimeric FGF19 polypeptide of  claim 1 , wherein the in vivo physiological half-life of the chimeric FGF19 polypeptide is at least or about the same as FGF19. 
     
     
         31 . The chimeric FGF19 polypeptide of  claim 1 , wherein the in vivo physiological half-life of the chimeric FGF19 polypeptide is at least or about the same as FGF21. 
     
     
         32 . A pharmaceutical composition comprising:
 (a) a therapeutically effective amount of the chimeric FGF19 polypeptide of  claim 1 ; and   (b) an acceptable pharmaceutical carrier.   
     
     
         33 . A method of treating an individual exhibiting one or more of obesity, type 1 diabetes, type 2 diabetes, high blood glucose, metabolic syndrome, atherosclerosis, hypercholesterolemia, stroke, osteoporosis, osteoarthritis, degenerative joint disease, muscle atrophy, sarcopenia, decreased lean body mass, baldness, wrinkles, increased fatigue, decreased stamina, decreased cardiac function, immune system dysfunction, cancer, Parkinson's disease, senile dementia, Alzheimer's disease and decreased cognitive function, the method comprising administering to the individual a therapeutically effective amount of the pharmaceutical composition of  claim 32 . 
     
     
         34 . A method of lowering the blood glucose of an individual in need of such treatment, the method comprising administering to the individual a therapeutically effective amount of the pharmaceutical composition of  claim 32 . 
     
     
         35 . The method of  claim 33 , wherein the individual is a human. 
     
     
         36 . An isolated nucleic acid encoding the chimeric FGF19 polypeptide of  claim 1 , or (b) the complement of the DNA molecule of (a). 
     
     
         37 - 40 . (canceled) 
     
     
         41 . The isolated nucleic acid of  claim 36 , wherein the encoded polypeptide further comprises the amino acid residues corresponding to the Fc portion of an immunoglobulin. 
     
     
         42 . An expression system comprising the nucleic acid molecule of  claim 36 . 
     
     
         43 . A host cell comprising the expression system of  claim 42 . 
     
     
         44 . A host cell comprising the nucleic acid molecule of  claim 36 . 
     
     
         45 . (canceled) 
     
     
         46 . A process for producing an isolated polypeptide comprising:
 culturing the host cell of  claim 44  under conditions suitable for expression of the encoded polypeptide; and recovering the encoded polypeptide from the cell culture.   
     
     
         47 . An isolated polypeptide produced by the process of  claim 46 . 
     
     
         48 . An antibody that binds specifically to any one of the chimeric FGF19 polypeptides of  claim 1 , wherein the antibody does not bind native FGF19 polypeptide or native FGF21 polypeptide. 
     
     
         49 . The antibody of  claim 48 , wherein the antibody is monoclonal. 
     
     
         50 . (canceled)