Human insulin analogues

Abstract

Human insulin analogs having a positively charged amino acid residue, i.e. Lys or Arg, in position B28, further being modified in the C-terminal end of the B-chain from Phe B24 to the C-terminal amino acid residue and wherein A18, A21 and/or B3 is different from Asn as well as insulin preparations containing the human insulin analogs are provided.

1 . Human insulin analogues, characterized in that they have a positively charged amino acid residue, i.e. Lys or Arg, in position B28, that they optionally are further modified in the C-terminal end of the B-chain from Phe B24  to the C-terminal amino acid residue, with the proviso that there is no Pro in Position B29, and that optionally one or more of the amino acid residues in positions A18, A21 and B3 are different from Asn with the proviso that it does not cover compounds protected by the patent originating from Ser. No. 07/453,445.  
     
     
         2 . Human insulin analogues, characterized in that they have the following formula:  
       
         
           
           
               
               
           
         
       
       wherein X 1 , X 2 , X 3 , X 5 , Y 1 , Y 2  and Y 3  are any naturally occurring amino acid residue, X 4  is Lys or Arg, X 6  is any naturally occurring amino acid residue carrying the C-terminal hydroxy group or hydroxy, or X 5  and X 6  together form the C-terminal hydroxy group with the proviso that it does not cover compounds protected by the patent originating from Ser. No. 07/453,445.  
     
     
         3 . Human insulin analogues according to  claim 2 , wherein X 5  is selected from the group consisting of any naturally occurring amino acid residue except Pro.  
     
     
         4 . Human insulin analogues according to  claim 2 , wherein one or more of Y 1 , Y 2  and Y 3 , independent of each other, is selected from the group consisting of any naturally occurring amino acid residue except Asn.  
     
     
         5 . Human insulin analogues according to  claim 2 , wherein 
 X 1  is selected from the group consisting of Phe, Ala, His, Thr, Ser, Asn and Tyr;    X 2  is selected from the group consisting of Tyr, Thr, Glu, Asp, Ala, His, Ser and Phe;    X 3  is selected from the group consisting of Pro, Glu, Asp, Ser, Thr and His;    X 5  is selected from the group consisting of Lys, Thr, Ser, Ala, Asp and Glu;    X 6  is selected from the group consisting of Thr-OH, Ser-OH, Ala-OH, Asp-OH, Glu-OH and hydroxy, or X 5  and X 6  together form the C-terminal hydroxy group;    Y 1  is selected from the group consisting of Asn, Asp, Gly, Ser, Glu and Ala,    Y 2  is selected from the group consisting of Asn, Gln, Glu, Asp and Thr, and    Y 3  is selected from the group consisting of Asn, Asp, Gly, Ser, Glu or Ala.    
     
     
         6 . Human insulin analogues according to  claim 2 , wherein 
 X 1  is selected from the group consisting of Phe, Ala, His, Thr, Ser, Asn or Tyr;    X 2  is selected from the group consisting of Tyr, Thr, Glu, Asp, Ala, His, Ser and Phe;    X 3  is selected from the group consisting of Pro, Glu, Asp, Ser, Thr and His;    X 5  is selected from the group consisting of Lys, Thr, Ser, Ala, Asp and Glu;    X 6  is hydroxy;    Y 1  is selected from the group consisting of Asn, Asp, Gly, Ser, Glu and Ala;    Y 2  is selected from the group consisting of Asn, Gin, Glu, Asp and Thr; and    Y 3  is selected from the group consisting of Asn, Asp, Gly, Ser, Glu and Ala.    
     
     
         7 . Human insulin analogues according to  claim 2 , wherein 
 X 1  is selected from the group consisting of Phe, Ala, His, Thr, Ser, Asn and Tyr;    X 2  is selected from the group consisting of Tyr, Thr, Glu, Asp, Ala, His, Ser and Phe;    X 3  is selected from the group consisting of Pro, Glu, Asp, Ser, Thr and His;    X 5  and X 6  together form the C-terminal hydroxy group;    Y 1  is selected from the group consisting of Asn, Asp, Gly, Glu, Ser and Ala;    Y 2  is selected from the group consisting of Asn, Gln, Glu, Asp and Thr; and    Y 3  is selected from the group consisting of Asn, Asp, Gly, Glu, Ser and Ala.    
     
     
         8 . Human insulin analogues according to  claim 2 , wherein X 1  is Phe; X 2  is Tyr; X 3  is Thr; X 5  is Lys; X 6  is Thr-OH or hydroxy; Y 1  is selected from the group consisting of Asn, Asp, Ser and Gly, Y 2  is selected from the group consisting of Asn, Gln, Asp, Glu and Thr, and Y 3  is selected from the group consisting of Asn, Asp, Ser and Gly.  
     
     
         9 . Human insulin analogues according to  claim 2 , wherein X 1  is Tyr; X 2  is Thr; X 3  is Pro; X 5  is Thr; X 6  is hydroxy; Y 1  is selected from the group consisting of Asn, Asp, Ser and Gly, Y 2  is selected from the group consisting of Asn, Gin, Asp, Glu and Thr, and Y 3  is selected from the group consisting of Asn, Asp, Ser and Gly.  
     
     
         10 . Human insulin analogues according to  claim 2 , wherein X 1  is Phe; X 2  is Thr; X 3  is Pro; X 5  is Thr; X 6  is hydroxy; Y 1  is selected from the group consisting of Asn, Asp, Ser and Gly, Y 2  is selected from the group consisting of Asn, Gin, Asp, Glu and Thr, and Y 3  is selected from the group consisting of Asn, Asp, Ser and Gly.  
     
     
         11 . Human insulin analogues according to  claim 2 , wherein X 1  is Phe; X 2  is Tyr; X 3  is Pro; X 5  is Thr; X 6  is hydroxy; Y 1  is selected from the group consisting of Asn, Asp, Ser and Gly, and Y 2  is selected from the group consisting of Asn, Gin, Asp, Glu and Thr, and Y 3  is selected from the group consisting of Asn, Asp, Ser and Gly.  
     
     
         12 . Human insulin analogues according to  claim 2 , wherein X 1  is Phe; X 2  is Tyr; X 3  is Thr; X 5  is Thr; X 6  is hydroxy; Y 1  is selected from the group consisting of Asn, Asp, Ser and Gly, and Y 2  is selected from the group consisting of Asn, Gin, Asp, Glu and Thr, and Y 3  is selected from the group consisting of Asn, Asp, Ser and Gly.  
     
     
         13 . Human insulin analogues according to  claim 2 , wherein said X 1  amino acid is uncharged and has a carbon atom in the gamma-position which is sp 2 -hybridized and X 6  is hydroxy.  
     
     
         14 . Human insulin analogues according to  claim 13 , wherein Y 1 , Y 2  and Y 3  are selected from the group consisting of any naturally occurring amino acid residue except Asn, and X 5  is selected from the group consisting of any naturally occurring amino acid residue except Thr.  
     
     
         15 . Human insulin analogues according to  claim 9 , wherein X 5  and X 6  together form hydroxy.  
     
     
         16 . Pharmaceutical composition comprising a human insulin analogue having the following formula:  
       
         
           
           
               
               
           
         
       
       wherein X 1 , X 2 , X 3 , X 5 , Y 1 , Y 2  and Y 3  are any naturally occurring amino acid residue, X 4  is Lys or Arg, X 6  is any naturally occurring amino acid residue carrying the C-terminal hydroxy group or hydroxy, or X 5  and X 6  together form the C-terminal hydroxy group.  
     
     
         17 . A method of treating Diabetes mellitus comprising providing to an effected individual a pharmaceutical composition comprising a human insulin analogue having the following formula:  
       
         
           
           
               
               
           
         
       
       wherein X 1 , X 2 , X 3 , X 5 , Y 1 , Y 2  and Y 3  are any naturally occurring amino acid residue, X 4  is Lys or Arg, X 6  is any naturally occurring amino acid residue carrying the C-terminal hydroxy group or hydroxy, or X 5  and X 6  together form the C-terminal hydroxy group, with the proviso that it does not cover compounds protected by the patent originating from Ser. No. 07/453,445, or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.  
     
     
         18 . The method according to claim  39 , wherein said insulin analogue exhibits low solubility at a pH value of 7.3.  
     
     
         19 . The method according to claim  40 , wherein said insulin analogue is essentially monomeric.  
     
     
         20 . The method according to claim  39 , wherein said pharmaceutical composition is formulated as an aqueous solution.