Novel insulin derivatives

Abstract

The present invention relates to insulin derivatives which are naturally occurring insulins or analogues thereof which have a side chain attached either to the α-amino group of the N-terminal amino acid residue of the B chain or to the ε-amino group of a Lys residue present in the B chain of the parent insulin, the side chain being of the general formula: —W—X—Y—Z wherein W, X, Y and Z are as defined in the disclosure.

1 . An insulin derivative which is a naturally occurring insulin or an analogue thereof which has a side chain attached to the α-amino group of the N-terminal amino acid residue of the B chain or to the ε-amino group of a Lys residue present in the B chain of the parent insulin, the side chain being of the general formula:
   —W—X—Y—Z
 
 wherein W is:
 an α-amino acid residue having a carboxylic acid group in the side chain which residue forms, with one of its carboxylic acid groups, an amide group together with the α-amino group of the N-terminal amino acid residue of the B chain or together with the ε-amino group of a Lys residue present in the B chain of the parent insulin; 
 a chain composed of two, three or four α-amino acid residues linked together via amide bonds, which chain—via an amide bond—is linked to the α-amino group of the N-terminal amino acid residue of the B chain or to the ε-amino group of a Lys residue present in the B chain of the parent insulin, the amino acid residues of W being selected from the group of amino acid residues having a neutral side chain and amino acid residues having a carboxylic acid group in the side chain so that W has at least one amino acid residue which has a carboxylic acid group in the side chain; or 
 a covalent bond from X to the α-amino group of the N-terminal amino acid residue of the B chain or to the ε-amino group of a Lys residue present in the B chain of the parent insulin; 
 
 X is:
 — C O—; 
 —CH(COOH) C O—; 
 —N(CH 2 COOH)CH 2   C O—; 
 —N(CH 2 COOH)CH 2 CON(CH 2 COOH)CH 2   C O—; 
 —N(CH 2 CH 2 COOH)CH 2 CH 2   C O—; 
 —N(CH 2 CH 2 COOH)CH 2 CH 2 CON(CH 2 CH 2 COOH)CH 2 CH 2   C O—; 
 —NHCH(COOH)(CH 2 ) 4 NH C O—; 
 —N(CH 2 CH 2 COOH)CH 2   C O—; or 
 —N(CH 2 COOH)CH 2 CH 2   C O—. 
 
 that 
 a) when W is an amino acid residue or a chain of amino acid residues, via a bond from the underscored carbonyl carbon forms an amide bond with an amino group in W, or 
 b) when W is a covalent bond, via a bond from the underscored carbonyl carbon forms an amide bond with the N-terminal α-amino group in the B chain or with the ε-amino group of a Lys residue present in the B chain of the parent insulin; 
 Y is:
 —(CH 2 ) m — where m is an integer in the range of 6 to 32; 
 a divalent hydrocarbon chain comprising 1, 2 or 3 CH═CH— groups and a number of —CH 2 — groups sufficient to give a total number of carbon atoms in the chain in the range of 10 to 32; 
 a divalent hydrocarbon chain of the formula —(CH 2 ) v C 6 H 4 (CH 2 ) w — wherein v and w are integers or one of them is zero so that the sum of v and w is in the range of 6 to 30; and 
 
 Z is:
 —COOH; 
 —CO-Asp; 
 —CO-Glu; 
 —CO-Gly; 
 —CO-Sar; 
 —CH(COOH) 2 ; 
 —N(CH 2 COOH) 2 ; 
 —SO 3 H; or 
 —PO 3 H; 
 
 and any Zn 2+  complexes thereof, provided that when W is a covalent bond and X is —CO—, then Z is different from —COOH. 
 
     
     
         2 . An insulin derivative according to  claim 1 , wherein side chain —W—X—Y—Z is attached to the α-amino group of the N-terminal amino acid residue of the B chain of the parent insulin. 
     
     
         3 . An insulin derivative according to  claim 1 , wherein side chain —W—X—Y—Z is attached to the ε-amino group of a Lys residue present in the B chain of the parent insulin. 
     
     
         4 . An insulin derivative according to  claim 1 , wherein W is a covalent bond. 
     
     
         5 . An insulin derivative according to  claim 1 , wherein W is an α-amino acid residue having from 4 to 10 carbon atoms. 
     
     
         6 . An insulin derivative according to  claim 5 , wherein W is selected from the group consisting of α-Asp, β-Asp, α-Glu, γ-Glu, α-hGlu and δ-hGlu. 
     
     
         7 . An insulin derivative according to  claim 1 , wherein W is a chain composed of two α-amino acid residues of which one has from 4 to 10 carbon atoms and a free carboxylic acid group while the other has from 2 to 11 carbon atoms but no free carboxylic acid group. 
     
     
         8 . An insulin derivative according to  claim 7  wherein W is selected from the group consisting of α-Asp-Gly; Gly-α-Asp; β-Asp-Gly; Gly-β-Asp; α-Glu-Gly; Gly-α-Glu; γ-Glu-Gly; Gly-γ-Glu; α-hGlu-Gly; Gly-α-hGlu; δ-hGlu-Gly; and Gly-δ-hGlu. 
     
     
         9 . An insulin derivative according to  claim 1 , wherein W is a chain composed of two α-amino acid residues, independently having from 4 to 10 carbon atoms, and both having a free carboxylic acid group. 
     
     
         10 . An insulin derivative according to  claim 9 , wherein W is selected from the group consisting of α-Asp-α-Asp; α-Asp-α-Glu; α-Asp-α-hGlu; α-Asp-β-Asp; α-Asp-γ-Glu; α-Asp-δ-hGlu; β-Asp-α-Asp; β-Asp-α-Glu; β-Asp-α-hGlu; β-Asp-β-Asp; β-Asp-γ-Glu; β-Asp-δ-hGlu; α-Glu-α-Asp; α-Glu-α-Glu; α-Glu-α-hGlu; α-Glu-β-Asp; α-Glu-γ-Glu; α-Glu-δ-hGlu; γ-Glu-α-Asp; γ-Glu-α-Glu; γ-Glu-α-hGlu; γ-Glu-β-Asp; γ-Glu-γ-Glu; γ-Glu-δ-hGlu; α-hGlu-α-Asp; α-hGlu-α-Glu; α-hGlu-α-hGlu; α-hGlu-β-Asp; α-hGlu-γ-Glu; α-hGlu-δ-hGlu; δ-hGlu-α-Asp; δ-hGlu-α-Glu; δ-hGlu-α-hGlu; δ-hGlu-β-Asp; δ-hGlu-γ-Glu; and δ-hGlu-δ-hGlu. 
     
     
         11 . An insulin derivative according to  claim 1 , wherein X is —CO— or —CH(COOH)CO—. 
     
     
         12 . An insulin derivative according to  claim 1 , wherein X is
 —N(CH 2 COOH)CH 2   C O—;   —N(CH 2 COOH)CH 2 CON(CH 2 COOH)CH 2   C O—;   —N(CH 2 CH 2 COOH)CH 2 CH 2   C O—;   —N(CH 2 CH 2 COOH)CH 2 CH 2 CON(CH 2 CH 2 COOH)CH 2 CH 2   C O—;   —N(CH 2 CH 2 COOH)CH 2   C O—; or   —N(CH 2 COOH)CH 2 CH 2   C O—.   
     
     
         13 . An insulin derivative according to  claim 1 , wherein Y is —(CH 2 ) m — where m is an integer in the range of from 6 to 32, from 8 to 20, from 12 to 20 or from 12-16. 
     
     
         14 . An insulin derivative according to  claim 1 , wherein Z is —COOH. 
     
     
         15 . An insulin derivative according to  claim 1 , wherein Z is —CH(COOH) 2 . 
     
     
         16 . An insulin derivative according to  claim 1 , wherein Z is —N(CH 2 COOH) 2 . 
     
     
         17 . An insulin derivative according to  claim 1 , wherein Z is —SO 3 H. 
     
     
         18 . An insulin derivative according to  claim 1 , wherein Z is —PO 3 H. 
     
     
         19 . An insulin derivative according to  claim 1 , wherein the parent insulin has Asn or Gly at position A21. 
     
     
         20 . An insulin derivative according to  claim 1 , wherein the parent insulin is a des(B1) analogue. 
     
     
         21 . An insulin derivative according to  claim 1 , wherein the parent insulin is a des(B30) analogue. 
     
     
         22 . An insulin derivative according to  claim 1 , wherein position B29 in the parent insulin can be any codable amino acid except Cys, Met, Arg and Lys and the amino acid in position B30 is Lys. 
     
     
         23 . An insulin derivative according to  claim 1 , wherein the parent insulin has Thr at position B29 and Lys at position B30. 
     
     
         24 . An insulin derivative according to  claim 1 , wherein the parent insulin is selected from the group consisting of human insulin; des(B1) human insulin; des(B30) human insulin; Gly A21  human insulin; Gly A21  des(B30) human insulin; Asp B28  human insulin; porcine insulin; Lys B28 Pro B29  human insulin; Gly A21 Arg B31 Arg B32  human insulin; and Lys B3 Glu B29  human insulin. 
     
     
         25 . An insulin derivative according to  claim 1  selected from the group consisting of N εB29 —(N α —(HOOC(CH 2 ) 14 CO)-γ-Glu) des(B30) human insulin; N εB29 —(N α —(HOOC(CH 2 ) 15 CO)-γ-Glu) des(B30) human insulin; N εB29 —(N α —(HOOC(CH 2 ) 16 CO)-γ-Glu) des(B30) human insulin; N εB29 —(N α —(HOOC(CH 2 ) 17 CO)-γ-Glu) des(B30) human insulin; N εB29 —(N α —(HOOC(CH 2 ) 18 CO)-γ-Glu) des(B30) human insulin; N εB29 —(N α —(HOOC(CH 2 ) 16 CO)-γ-Glu-N-(γ-Glu)) des(B30) human insulin; N εB29 (Na-(Asp-OC(CH 2 ) 16 CO)-γ-Glu) des(B30) human insulin; N εB29 —(N α -(Glu-OC(CH 2 ) 14 CO)-γ-Glu) des(B30) human insulin; N εB29 —(N α -(Glu-OC(CH 2 ) 14 CO—) des(B30) human insulin; N εB29 —(N α -(Asp-OC(CH 2 ) 16 CO—) des(B30) human insulin; N εB29 (Na—(HOOC(CH 2 ) 16 CO)-α-Glu-N-(β-Asp)) des(B30) human insulin; N εB29 —(N α -(Gly-OC(CH 2 ) 13 CO)-γ-Glu) des(B30) human insulin; N εB29 —(N α -(Sar-OC(CH 2 ) 13 CO)-γ-Glu) des(B30) human insulin; N εB29 —(N α —(HOOC(CH 2 ) 13 CO)-γ-Glu) des(B30) human insulin; (N εB29 (Na—(HOOC(CH 2 ) 13 CO) 43 -Asp) des(B30) human insulin; N εB29 —(N α —(HOOC(CH 2 ) 13 CO)-α-Glu) des(B30) human insulin; N εB29 —(N α —(HOOC(CH 2 ) 16 CO)-γ-D-Glu) des(B30) human insulin; N εB29 —(N α —(HOOC(CH 2 ) 14 CO)-γ-D-Asp) des(B30) human insulin N εB29 —(N α —(HOOC(CH 2 ) 14 CO)-γ-D-Asp) des(B30) human insulin; N εB29 —(N—HOOC(CH 2 ) 16 CO-γ-D-Asp) des(B30) human insulin; N εB29 —(N—HOOC(CH 2 ) 14 CO-IDA) des(B30) human insulin; N εB29 —[N—(HOOC(CH 2 ) 16 CO)—N-(carboxyethyl)-Gly] des(B30) human insulin; N εB29 —[N—(HOOC(CH 2 ) 14 CO)—N-(carboxyethyl)-Gly] des(B30) human insulin; and N εB29 —[N—(HOOC(CH 2 ) 14 CO)—N-(carboxymethyl)-β-Ala] des(B30) human insulin. 
     
     
         26 . A zinc complex of an insulin derivative according to  claim 1 , wherein each insulin hexamer in said complex binds two zinc ions. 
     
     
         27 . A zinc complex of an insulin derivative according to  claim 1 , wherein each insulin hexamer in said complex binds three zinc ions. 
     
     
         28 . A zinc complex of an insulin derivative according to  claim 1 , wherein each insulin hexamer in said complex binds four zinc ions. 
     
     
         29 . A pharmaceutical composition for the treatment of diabetes in a patient in need of such treatment, said composition comprising a therapeutically effective amount of an insulin derivative according to  claim 1  together with a pharmaceutically acceptable carrier. 
     
     
         30 . A pharmaceutical composition for the treatment of diabetes in a patient in need of such treatment, said composition comprising a therapeutically effective amount of an insulin derivative according to  claim 1  in mixture with an insulin or an insulin analogue which has a rapid onset of action, together with a pharmaceutically acceptable carrier. 
     
     
         31 . A method of treating diabetes, said method comprising administering to a patient in need of such a treatment a therapeutically effective amount of an insulin derivative according to  claim 1  together with a pharmaceutically acceptable carrier. 
     
     
         32 . A method of treating diabetes, said method comprising administering to a patient in need of such a treatment a therapeutically effective amount of an insulin derivative according to  claim 1  in mixture with an insulin or an insulin analogue which has a rapid onset of action, together with a pharmaceutically acceptable carrier.