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 B1 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 ε-amino group of Lys B29 ; 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 ε-amino group of Lys B29 has a lipophilic substituent; and any Zn 2+ complexes thereof with the proviso that when B30 is Thr or Ala and A21 and B3 are both Asn, and Phe B1 is present, then the insulin derivative is always present as a Zn 2+ complex.

1  An insulin derivative having the following sequence:  
       
         
           
           
               
               
           
         
       
       wherein 
 (a) Aaa at positions A21 and B3 are, independently, any amino acid residue which can be coded for by the genetic code except Lys, Arg and Cys;  
 (b) Xaa at position B1 is Phe or is deleted;  
 (c) Xaa at position B30 is any amino acid residue which can be coded for by the genetic code except Lys, Arg and Cys; and  
 (d) the ε-amino group of Lys B29  is substituted with a lipophilic substituent having at least 10 carbon atoms;  
 wherein the insulin derivative is a Zn 2+  complex and the Zn 2+  complex of the insulin derivative is more water soluble than the insulin derivative without Zn 2+ .  
 
     
     
         2  The insulin derivative according to  claim 1 , wherein Xaa at position A21 is Asn.  
     
     
         3  The insulin derivative according to  claim 2 , wherein the lipophilic substituent has from 12 to 24 carbon atoms.  
     
     
         4  The insulin derivative according to  claim 1 , wherein Xaa at position A21 is Ala, Asn, Gin, Gly or Ser.  
     
     
         5  The insulin derivative according to  claim 4 , wherein the lipophilic substituent has from 12 to 24 carbon atoms.  
     
     
         6  The insulin derivative according to  claim 1 , wherein Xaa at position B1 is deleted.  
     
     
         7  The insulin derivative according to  claim 6 , wherein the lipophilic substituent has from 12 to 24 carbon atoms.  
     
     
         8  The insulin derivative according to  claim 1 , wherein Xaa at position B1 is Phe.  
     
     
         9  The insulin derivative according to  claim 8 , wherein the lipophilic substituent has from 12 to 24 carbon atoms.  
     
     
         10  The insulin derivative according to  claim 1 , wherein Xaa at position B3 is Asn, Asp, Gln or Thr.  
     
     
         11  The insulin derivative according to  claim 10 , wherein the lipophilic substituent has from 12 to 24 carbon atoms.  
     
     
         12  The insulin derivative according to  claim 1 , wherein Xaa at position B30 is Ala or Thr.  
     
     
         13  The insulin derivative according to  claim 12 , wherein the lipophilic substituent has from 12 to 24 carbon atoms.  
     
     
         14  The insulin derivative according to  claim 1 , wherein Xaa at position A21 is Ala, Asn, Gin, Gly or Ser, Xaa at position B3 is Asn, Asp, Gin or Thr, and Xaa at position B30 is Ala or Thr.  
     
     
         15  The insulin derivative according to  claim 14 , wherein the lipophilic substituent has from 12 to 24 carbon atoms.  
     
     
         16  The insulin derivative according to  claim 1 , wherein Xaa at position A21 is Asn, Xaa at position B3 is Asn, Xaa at position B1 is Phe and Xaa at position B30 is Thr.  
     
     
         17  The insulin derivative according to  claim 16 , wherein the lipophilic substituent has from 12 to 24 carbon atoms.  
     
     
         18  The insulin derivative according to  claim 1 , wherein the lipophilic substituent has from 12 to 24 carbon atoms.  
     
     
         19  The insulin derivative according to  claim 1  which is in the form of a hexamer.  
     
     
         20  The insulin derivative according to  claim 19 , wherein the lipophilic substituent has from 12 to 24 carbon atoms.  
     
     
         21  The insulin derivative according to  claim 19 , wherein Xaa at position A21 is Asn, Xaa at position B1 is Phe, Xaa at position B3 is Asn, and Xaa at position B30 is Thr.  
     
     
         22  The insulin derivative according to  claim 19 , wherein two zinc ions bind to the hexamer.  
     
     
         23  The insulin derivative according to  claim 22 , wherein the lipophilic substituent has from 12 to 24 carbon atoms.  
     
     
         24  The insulin derivative according to  claim 19 , wherein three zinc ions bind to the hexamer.  
     
     
         25  The insulin derivative according to  claim 24 , wherein the lipophilic substituent has from 12 to 24 carbon atoms.  
     
     
         26  The insulin derivative according to  claim 19 , wherein four zinc ions bind to the hexamer.  
     
     
         27  The insulin derivative according to  claim 26 , wherein the lipophilic substituent has from 12 to 24 carbon atoms.  
     
     
         28  A pharmaceutical composition which is an aqueous solution, comprising (a) an insulin derivative according to  claim 1 , (b) an isotonic agent, (c) a preservative and (d) a buffer.  
     
     
         29  The pharmaceutical composition according to  claim 28 , wherein the pH of the aqueous solution is in the range of 6.5-8.5.  
     
     
         30  The pharmaceutical composition according to  claim 28 , wherein the solubility of the insulin derivative exceeds 600 nmol/ml of the aqueous solution.  
     
     
         31  The pharmaceutical composition according to  claim 28 , further comprising an insulin or an insulin analogue which has a rapid onset of action.  
     
     
         32  The pharmaceutical composition according to  claim 28 , wherein Xaa at position A21 is Asn, Xaa at position B3 is Asn, Xaa at position B1 is Phe and Xaa at position B30 is Thr.  
     
     
         33  The pharmaceutical composition according to  claim 28 , wherein the lipophilic substituent has from 12 to 24 carbon atoms.  
     
     
         34  The pharmaceutical composition according to  claim 28 , wherein the insulin derivative is in the form of a hexamer.  
     
     
         35  A method of treating diabetes in a patient in need of such a treatment, comprising administering to the patient a therapeutically effective amount of a pharmaceutical composition according to  claim 28 .  
     
     
         36  An insulin derivative having the following sequence:  
       
         
           
           
               
               
           
         
       
       wherein 
 (a) Xaa at positions A21 and B3 are, independently, any amino acid residue which can be coded for by the genetic code except Lys, Arg and Cys;  
 (b) Xaa at position BI is Phe or is deleted;  
 (c) Xaa at position B30 is deleted; and  
 (d) the ε-amiiino group of Lys B29  is substituted with a lipophilic substituent having at least 10 carbon atoms;  
 wherein the insulin derivative is a Zn 2+  complex and the Zn 2+  complex of the insulin derivative is more water soluble than the insulin derivative without Zn 2+ .  
 
     
     
         37  The insulin derivative according to  claim 36 , wherein Xaa at position A21 is Ala, Asn, Gln, Gly or Ser.  
     
     
         38  The insulin derivative according to  claim 37 , wherein the lipophilic substituent has from 12 to 24 carbon atoms.  
     
     
         39  The insulin derivative according to  claim 36 , wherein Xaa at position B1 is deleted.  
     
     
         40  The insulin derivative according to  claim 39 , wherein the lipophilic substituent has from 12 to 24 carbon atoms.  
     
     
         41  Thc insulin derivative according to  claim 36 , wherein Xaa at position B1 is Phe.  
     
     
         42  The insulin derivative according to  claim 41 , wherein the lipophilic substituent has from 12 to 24 carbon atoms.  
     
     
         43  The insulin derivative according to  claim 36 , wherein Xaa at position B3 is Asn, Asp, Gln or Thr.  
     
     
         44  The insulin derivative according to  claim 43 , wherein the lipophilic substituent has from 12 to 24 carbon atoms.  
     
     
         45  The insulin derivative according to  claim 36  wherein Xaa at position A21 is Ala, Asn, Gln, Gly or Ser, and Xaa at position B3 is Asn, Asp, Gin or Thr.  
     
     
         46  The insulin derivative according to  claim 45 , wherein the lipophilic substituent has from 12 to 24 carbon atoms.  
     
     
         47  The insulin derivative according to  claim 36 , wherein Xaa at position A21 is Asn, Xaa at position B1 is Phe, and Xaa at position B3 is Asn.  
     
     
         48  The insulin derivative according to  claim 47 , wherein the lipophilic substituent has from 12 to 24 carbon atoms.  
     
     
         49  The insulin derivative according to  claim 36 , wherein the lipophilic substituent has from 12 to 24 carbon atoms.  
     
     
         50  The insulin derivative according to  claim 36  which is in the form of a hexamer.  
     
     
         51  The insulin derivative according to  claim 50 , wherein the lipophilic substituent has from 12 to 24 carbon atoms.  
     
     
         52  The insulin derivative according to  claim 50 , wherein Xaa at position A21 is Asn, Xaa at position B3 is Asn, and Xaa at position B1 is Phe.  
     
     
         53  The insulin derivative according to  claim 50 , wherein two zinc ions bind to the hexamer.  
     
     
         54  The insulin derivative according to  claim 53 , wherein the lipophilic substituent has from 12 to 24 carbon atoms.  
     
     
         55  The insulin derivative according to  claim 50 , wherein three zinc ions bind to the hexamer.  
     
     
         56  The insulin derivative according to  claim 55 , wherein the lipophilic substituent has from 12 to 24 carbon atoms.  
     
     
         57  The insulin derivative according to  claim 50 , wherein four zinc ions bind to the hexamer.  
     
     
         58  The insulin derivative according to  claim 57 , wherein the lipophilic substituent has from 12 to 24 carbon atoms.  
     
     
         59  A pharmaceutical composition which is an aqueous solution, comprising (a) an insulin derivative according to  claim 36 , (b) an isotonic agent, (c) a preservative and (d) a buffer.  
     
     
         60  The pharmaceutical composition according to  claim 59 , wherein the pH of the aqueous solution is in the range of 6.5-8.5.  
     
     
         61  The pharmaceutical composition according to  claim 59 , wherein the solubility of the insulin derivative exceeds 600 nmol/ml of the aqueous solution.  
     
     
         62  The pharmaceutical composition according to  claim 59 , further comprising an insulin or an insulin analogue which has a rapid onset of action.  
     
     
         63  The pharmaceutical composition according to  claim 59 , wherein the insulin derivative is a Zn 2+  complex.  
     
     
         64  The pharmaceutical composition according to  claim 59 , wherein Xaa at position A21 is Asn, Xaa at position B3 is Asn, and Xaa at position B1 is Phe.  
     
     
         65  The pharmaceutical composition according to  claim 59 , wherein the lipophilic substituent has from 12 to 24 carbon atoms.  
     
     
         66  The pharmaceutical composition according to  claim 59 , wherein the insulin derivative is in the form of a hexamer.  
     
     
         67  A method of treating diabetes in a patient in need of such a treatment, comprising administering to the patient a therapeutically effective amount of a pharmaceutical composition according to  claim 59.