Acylated insulin

Abstract

The present invention relates to protracted human insulin derivatives in which the A21 and the B3 amino acid residues are, independently, any amino acid residue which can be coded for by the genetic code except Lys, Arg and Cys; Phe<SUP>B1 </SUP>may be deleted; the B30 amino acid residue is (a) a non-codable, lipophilic amino acid having from 10 to 24 carbon atoms, in which case an acyl group of a carboxylic acid with up to 5 carbon atoms is bound to the epsilon-amino group of Lys<SUP>B29</SUP>; or (b) the B30 amino acid residue is deleted or is any amino acid residue which can be coded for by the genetic code except Lys, Arg and Cys, in any of which cases the C-amino group of Lys<SUP>B29 </SUP>has a lipophilic substituent; and any Zn<SUP>2+</SUP> complexes thereof with the proviso that when B30 is Thr or Ala and A21 and B3 are both Asn, and Phe<SUP>B1 </SUP>is present, then the insulin derivative is always present as a Zn<SUP>2+</SUP> complex.

1 . An insulin derivative having the following sequence:  
       
         
           
                 
               
                     
                 
                     
                 
                     
                 
                   
                     
                       
                       
                           
                           
                       
                     
                   
                 
                     
                 
                     
                 
             
                
                
               
               
                
                
                
                
               
            
           
         
       
       wherein 
 (a) Xaa at positions A21 and B3 are Asn;  
 (b) Xaa at position B1 is Phe;  
 (c) Xaa at position B30 is Thr; and  
 (d) the ε-amino group of Lys B29  is substituted with a lipophilic substituent having at least 6 carbon atoms;  
 wherein the insulin derivative is a Zn 2+  complex.  
 
     
     
         2 . The insulin derivative of  claim 1 , wherein the lipophilic substituent is an acyl group corresponding to a carboxylic acid having at least 6 carbon atoms.  
     
     
         3 . The insulin derivative of  claim 1 , wherein the lipophilic substituent is an acyl group corresponding to a fatty acid having at least 6 carbon atoms.  
     
     
         4 . The insulin derivative of  claim 1 , wherein 2-4 Zn 2+  ions are bound to each hexamer of the insulin derivative.  
     
     
         5 . The insulin derivative of  claim 1 , wherein the insulin derivative has a lipophilicity relative to human insulin, k′ rel , greater than 1.  
     
     
         6 . A pharmaceutical composition comprising (a) the insulin derivative of  claim 1 , (b) an isotonic agent, (c) a preservative, and (d) a buffer.  
     
     
         7 . The pharmaceutical composition of  claim 6 , said composition further comprising an insulin or an insulin analogue which has a rapid onset of action.  
     
     
         8 . The pharmaceutical composition of  claim 6 , wherein the insulin derivative is in the form of a hexamer.  
     
     
         9 . The insulin derivative of  claim 3 , wherein said derivative is N εB29 -octanoyl insulin.  
     
     
         10 . The insulin derivative of  claim 9 , wherein 2-4 Zn 2+  ions are bound to each hexamer of the insulin derivative.  
     
     
         11 . A pharmaceutical composition comprising (a) the insulin derivative of  claim 9 , (b) an isotonic agent, (c) a preservative, and (d) a buffer.  
     
     
         12 . The pharmaceutical composition of  claim 11 , wherein said composition is an aqueous solution.  
     
     
         13 . The pharmaceutical composition of  claim 12 , wherein the aqueous solution has a pH in the range of 6.5-8.5.  
     
     
         14 . The pharmaceutical composition of  claim 12 , wherein the insulin derivative has a solubility exceeding 600 nmol per ml of the aqueous solution.  
     
     
         15 . The pharmaceutical composition of  claim 11 , said composition further comprising an insulin or an insulin analogue which has a rapid onset of action.  
     
     
         16 . The pharmaceutical composition of  claim 11 , wherein the insulin derivative is in the form of a hexamer.  
     
     
         17 . The pharmaceutical composition of  claim 6 , wherein said composition is an aqueous solution.  
     
     
         18 . The pharmaceutical composition of  claim 17 , wherein the aqueous solution has a pH in the range of 6.5-8.5.  
     
     
         19 . The pharmaceutical composition of  claim 17 , wherein the insulin derivative has a solubility exceeding 600 nmol per ml of the aqueous solution.