Long-acting fgf21 fusion proteins and pharmaceutical composition comprising same

Abstract

A fusion protein comprises an FGF21 mutant protein and an Fc region of an immunoglobulin. The fusion protein exhibits improved pharmacological efficacy, in vivo duration and protein stability. A pharmaceutical composition containing the fusion protein as an active ingredient may be effectively used as a therapeutic agent for diabetes, obesity, dyslipidemia, metabolic syndrome, non-alcoholic fatty liver disease or non-alcoholic steatohepatitis.

1 . A fusion protein comprising a fibroblast growth factor 21 (FGF21) mutant protein and an Fc region of an immunoglobulin,
 wherein the FGF21 mutant protein comprises the following mutation:   (i) a substitution of the amino acid at position 180 from the N-terminus of a wild-type FGF21 protein with the amino acid E.   
     
     
         2 . The fusion protein of  claim 1 , wherein the FGF21 mutant protein further comprises the following mutation:
 (ii) a substitution of the amino acids at positions 170 to 174 from the N-terminus of a wild-type FGF21 protein with the amino acid sequence of TGLEAN (SEQ ID NO: 44).   
     
     
         3 . The fusion protein of  claim 1 , wherein the FGF21 mutant protein further comprises the following mutation:
 (iii) a substitution of the amino acid at position 170 from the N-terminus of a wild-type FGF21 protein with the amino acid N.   
     
     
         4 . The fusion protein of  claim 1 , wherein the FGF21 mutant protein further comprises the following mutations:
 (iv) a substitution of the amino acids at positions 98 to 101 from the N-terminus of a wild-type FGF21 protein with the amino acid sequence of EIRP (SEQ ID NO: 42);   (v) a substitution of the amino acids at positions 170 to 174 from the N-terminus of a wild-type FGF21 protein with the amino acid sequence of TGLEAV (SEQ ID NO: 43);   (vi) a substitution of the amino acid at position 174 from the N-terminus of a wild-type FGF21 protein with the amino acid N; or   (vii) a combination of the substitution (iv) and the substitution (v), or a combination of the substitution (iv) and the substitution (v).   
     
     
         5 . The fusion protein of  claim 1 , wherein the amino acid residue N of the FGF21 mutant protein introduced by a mutation is glycosylated. 
     
     
         6 . The fusion protein of  claim 1 , wherein the wild-type FGF21 protein comprises the amino acid sequence of SEQ ID NO: 1. 
     
     
         7 . The fusion protein of  claim 1 , wherein the FGF21 mutant protein comprises an amino acid sequence selected from the group consisting of SEQ ID NOs: 15 to 23. 
     
     
         8 . The fusion protein of  claim 1 , wherein the FGF21 mutant protein is connected to the Fc region of the immunoglobulin via a linker. 
     
     
         9 . The fusion protein of  claim 5 , wherein the linker is connected to the C-terminus of the Fc region of the immunoglobulin and the N-terminus of the FGF21 mutant protein. 
     
     
         10 . The fusion protein of  claim 5 , wherein the linker is a peptide consisting of 10 to 30 amino acid residues, and wherein the linker comprise an amino acid sequence selected from the group consisting of SEQ ID NOs: 2 to 5. 
     
     
         11 . The fusion protein of  claim 1 , wherein the Fc region of the immunoglobulin is any one of the Fc regions of IgG1, IgG2, IgG3, IgG4 and IgD, or a hybrid Fc containing a combination thereof. 
     
     
         12 . The fusion protein of  claim 9 , wherein the hybrid Fc comprises an IgG4 region and an IgD region. 
     
     
         13 . A pharmaceutical composition comprising the fusion protein according to  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         14 . An isolated nucleic acid molecule encoding the fusion protein according to  claim 1 . 
     
     
         15 . An expression vector comprising the nucleic acid molecule of  claim 14 . 
     
     
         16 . A host cell comprising the nucleic acid molecule of  claim 14 . 
     
     
         17 . A method for treating diabetes, obesity, dyslipidemia, metabolic syndrome, non-alcoholic fatty liver disease or non-alcoholic steatohepatitis in a subject in need thereof, comprising administering the pharmaceutical composition of  claim 13  to the subject. 
     
     
         18 . A method selected from the group consisting of:
 reducing blood glucose level in a subject;   reducing body weight in a subject;   reducing triglyceride or low-density lipoprotein levels in a subject; and   improving insulin sensitivity in a subject   wherein the method comprises administering the pharmaceutical composition of  claim 14  to the subject.   
     
     
         19 . The method of  claim 18 , wherein the subject is obese. 
     
     
         20 . The method of  claim 18 , wherein the subject has diabetes.