Glucagon analogues

Abstract

The invention provides materials and methods for promoting weight loss or preventing weight gain, and in the treatment of diabetes, metabolic syndrome and associated disorders. In particular, the invention provides novel glucagon analogue peptides effective in such methods. The peptides may mediate their effect by having increased selectivity for the GLP-1 receptor as compared to human glucagon.

1 . A nucleic acid encoding a peptide having the formula X-Z
 wherein   X is a peptide which has the formula I   
       
         
           
                 
               
                   (SEQ ID NO: 4) 
                 
                   His-Ser-Gln-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Lys- 
                 
                     
                 
                   Tyr-Leu-Asp-Arg-Ala-Arg-Ala-Asp-Asp-Phe-Val-Ala- 
                 
                     
                 
                   Trp-Leu-Lys-Glu-Ala 
                 
             
                
                
                
                
                
                
               
            
           
         
         or differs from formula I at up to 4 of the following positions whereby, if different from formula I: 
         the residue at position 2 is selected from: Aib, D-Ser; 
         the residue at position 16 is: Lys, Asp, Glu; 
         the residue at position 18 is selected from: Lys, H is, Ala, Ser, Tyr; 
         the residue at position 20 is selected from: Gln, H is, Lys, Arg, Glu; 
         the residue at position 21 is: Glu; 
         the residue at position 24 is selected from: Gln, Leu, Glu, Lys, Arg, Asp; 
         the residue at position 27 is selected from: Met, Cys, Arg, Glu, Leu; 
         the residue at position 28 is selected from: Asn, Ser, Arg, Lys, Ala, Leu, Glu, Asp; and 
         the residue at position 29 is selected from: Thr, Glu, Lys; 
         and Z is absent or a sequence of 1-20 amino acid units selected from the group consisting of Ala, Leu, Ser, Thr, Tyr, Cys, Glu, Lys, Arg, Dbu, Dpr and Orn; 
         with the proviso that X is not His-Ser-Gln-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Lys-Tyr-Leu-Asp-Arg-Ala-Arg-Ala-Asp-Asp-Phe-Val-Ala-Trp-Leu-Lys-Ser-Thr (SEQ ID NO:5). 
       
     
     
         2 . The nucleic acid according to  claim 1 , wherein X differs from formula I at up to 4 of the following positions whereby, if different from formula I:
 the residue at position 2 is selected from: Aib, D-Ser;   the residue at position 18 is selected from: Lys, H is, Ala, Ser, Tyr;   the residue at position 20 is selected from: Gln, H is, Lys, Arg, Glu;   the residue at position 24 is selected from: Gln, Leu, Glu, Lys, Arg;   the residue at position 27 is selected from: Met, Cys, Arg, Glu, Leu;   the residue at position 28 is selected from: Asn, Ser, Arg, Lys, Ala, Leu; and   the residue at position 29 is selected from: Thr, Glu, Lys.   
     
     
         3 . The nucleic acid according to  claim 2 , comprising the residues 27-Lys and 28-Ser. 
     
     
         4 . The nucleic acid according to  claim 3 , wherein X additionally differs from formula I at one or two of the following positions whereby, if different from formula I:
 the residue at position 2 is selected from: Aib, D-Ser;   the residue at position 18 is selected from: Lys, H is, Ala, Ser, Tyr;   the residue at position 20 is selected from: Gln, H is, Lys, Arg, Glu;   the residue at position 24 is selected from: Gln, Leu, Glu, Lys, Arg; and   the residue at position 29 is selected from: Thr, Glu, Lys.   
     
     
         5 . The nucleic acid according to  claim 1 , wherein the residues at positions 16 and 20 are capable of forming a salt bridge. 
     
     
         6 . The nucleic acid according to  claim 5 , comprising the residues 16-Arg, 20-Asp. 
     
     
         7 . The nucleic acid according to  claim 1 , wherein X comprises one or more of the following sets of residues:
 16-Arg;   16-Arg, 20-Asp;   16-Arg, 20-Asp, 24-Ala;   16-Arg, 20-Asp, 27-Lys, 28-Ser;   16-Arg, 20-Asp, 29-Ala;   16-Arg, 27-Lys, 28-Ser;   16-Arg, 27-Lys, 28-Ser, 29-Ala;   24-Ala, 27-Lys, 28-Ser;   24-Ala, 27-Lys, 28-Ser, 29-Ala;   24-Ala;   27-Lys; 28-Ser;   20-Glu, 28-Ser, 29-Thr;   24-Glu, 28-Ser, 29-Thr;   27-Glu, 28-Arg;   2-D-Ser, 28-Ser, 29-Thr; or   20-His, 28-Ser, 29-Thr.   
     
     
         8 . The nucleic acid according to  claim 1 , wherein X has the sequence: 
       
         
           
                 
                 
               
                     
                   (SEQ ID NO: 6) 
                 
                     
                   HSQGTFTSDYSKYLDRARADDFVAWLKSA; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 7) 
                 
                     
                   HSQGTFTSDYSKYLDRARADDFVAWLKEA; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 8) 
                 
                     
                   HSQGTFTSDYSKYLDRARAEDFVAWLKST; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 9) 
                 
                     
                   HSQGTFTSDYSKYLDRARADDFVEWLKST; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 10) 
                 
                     
                   HSQGTFTSDYSKYLDRARADDFVAWLERA; 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 11) 
                 
                     
                   H-D-Ser-QGTFTSDYSKYLDRARADDFVAWLKST; 
                 
                     
                   or 
                 
                     
                     
                 
                     
                   (SEQ ID NO: 12) 
                 
                     
                   HSQGTFTSDYSKYLDRARAHDFVAWLKST. 
                 
             
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
                
               
            
           
         
       
     
     
         9 . The nucleic acid according to  claim 1 , wherein Z has no more than 25% sequence identity with the corresponding portion of the IP-1 sequence of human oxyntomodulin having the sequence Lys-Arg-Asn-Arg-Asn-Asn-Ile-Ala (SEQ ID NO:2). 
     
     
         10 . The nucleic acid according to  claim 1 , wherein Z has a Cys as the C-terminal residue. 
     
     
         11 . The nucleic acid according to  claim 1 , wherein Z is absent. 
     
     
         12 . An expression vector comprising the nucleic acid according to  claim 1 . 
     
     
         13 . A host cell comprising the nucleic acid according to  claim 1 . 
     
     
         14 . A host cell comprising the expression vector according to  claim 12 . 
     
     
         15 . A pharmaceutical composition comprising the nucleic acid according to  claim 1 , in admixture with a pharmaceutically acceptable carrier. 
     
     
         16 . A pharmaceutical composition comprising the expression vector according to  claim 12 , in admixture with a pharmaceutically acceptable carrier. 
     
     
         17 . A pharmaceutical composition comprising the host cell according to  claim 13 , in admixture with a pharmaceutically acceptable carrier. 
     
     
         18 . A pharmaceutical composition comprising the host cell according to  claim 14 , in admixture with a pharmaceutically acceptable carrier. 
     
     
         19 . A method of reducing weight gain, promoting weight loss, or for treatment of a condition caused by or associated with excess body weight or obesity including morbid obesity, obesity linked inflammation, obesity linked gallbladder disease and obesity induced sleep apnea, or for treatment of insulin resistance, glucose intolerance, type 2 diabetes, hypertension, atherogenic dyslipidimia, atherosclerois, arteriosclerosis, coronary heart disease or stroke in a subject in need thereof, said method comprising administering the nucleic acid according to  claim 1  to said subject. 
     
     
         20 . A method of reducing weight gain, promoting weight loss, or for treatment of a condition caused by or associated with excess body weight or obesity including morbid obesity, obesity linked inflammation, obesity linked gallbladder disease and obesity induced sleep apnea, or for treatment of insulin resistance, glucose intolerance, type 2 diabetes, hypertension, atherogenic dyslipidimia, atherosclerois, arteriosclerosis, coronary heart disease or stroke in a subject in need thereof, said method comprising administering the expression vector according to  claim 12  to said subject. 
     
     
         21 . A method of reducing weight gain, promoting weight loss, or for treatment of a condition caused by or associated with excess body weight or obesity including morbid obesity, obesity linked inflammation, obesity linked gallbladder disease and obesity induced sleep apnea, or for treatment of insulin resistance, glucose intolerance, type 2 diabetes, hypertension, atherogenic dyslipidimia, atherosclerois, arteriosclerosis, coronary heart disease or stroke in a subject in need thereof, said method comprising administering the host cell according to  claim 13  to said subject. 
     
     
         22 . A method of reducing weight gain, promoting weight loss, or for treatment of a condition caused by or associated with excess body weight or obesity including morbid obesity, obesity linked inflammation, obesity linked gallbladder disease and obesity induced sleep apnea, or for treatment of insulin resistance, glucose intolerance, type 2 diabetes, hypertension, atherogenic dyslipidimia, atherosclerois, arteriosclerosis, coronary heart disease or stroke in a subject in need thereof, said method comprising administering the host cell according to  claim 14  to said subject.