Nitrogen-Based Linkers for Attaching Modifying Groups to Polypeptides and Other Macromolecules

Abstract

Compounds comprising a peptide moiety, a linker moiety and a water-soluble polymer moiety such as a poly(ethylene glycol) moiety are disclosed. Various linker moieties for use in these compounds are also disclosed, along methods for their synthesis.

1 . A linker moiety compound, wherein the linker moiety compound has 
       
         
           
           
               
               
           
         
       
       wherein
 α is an integer, 1≦α≦7; 
 β is an integer, 1≦β≦7; 
 δ is an integer, 2≦δ≦5; 
 A is either CO 2 H, activated CO 2 H, NH 2 , NCO, CHO, maleimide, or vinyl sulfone; and 
 E is either NH 2 , CO 2 H, CHO, maleimide, or NHBoc. 
 
     
     
         2 . The linker moiety compound of  claim 1 , wherein
 α=β=1 or 2;   δ=3;   A is either CO 2 H or activated CO 2 H; and   E is NHBoc.   
     
     
         3 . A linker moiety compound, wherein the linker moiety compound has the structure: 
       
         
           
           
               
               
           
         
       
       wherein
 κ is an integer, 0≦κ≦8; 
 ρ is an integer, 0≦ρ≦8; 
 τ is an integer, 2≦τ≦5; 
 A is either NHR or NRBoc; 
 R is alkyl; and 
 E is either NH 2 , CO 2 H, activated CO 2 H, CHO, maleimide, or NRBoc, wherein R is H, or alkyl. 
 
     
     
         4 . The linker moiety compound of  claim 3 , wherein
 κ=ρ=0;   τ=3;   A is NRBoc;   R is CH 3 ; and   E is CO 2 H or CONHS.   
     
     
         5 . A linker moiety compound, wherein the linker moiety compound has the structure: 
       
         
           
           
               
               
           
         
         wherein 
         χ is an integer, 1≦χ≦4 
         φ is an integer, 1≦φ≦4; 
         γ is an integer, 2≦γ≦8; 
         η is an integer 2≦η≦8; 
         A is either CO 2 H, activated CO 2 H, NH 2 , NCO, CHO, maleimide, or vinyl sulfone; 
         B is either CH or N; 
         C is either CO(CH 2 ) σ CO or (CH 2 ) σ ; 
         D is either CH or N; 
         E is either NH 2 , NHBoc, CO 2 H, CHO, or maleimide; and 
         σ is an integer, 2≦σ≦5. 
       
     
     
         6 . The linker moiety compound of  claim 5 , wherein
 χ=1;   φ=1;   γ is an integer 2≦γ≦3;   η is an integer 2≦η≦3;   A is either CO 2 H or activated CO 2 H;   B is N;   C is CO(CH 2 ) σ CO or (CH 2 ) σ ;   D is N;   E is NHBoc; and   σ is an integer, 2≦σ≦3.   
     
     
         7 - 8 . (canceled) 
     
     
         9 . A compound comprising a peptide moiety, a linker moiety, and a water-soluble polymer moiety, wherein the linker moiety is between the peptide moiety and the water-soluble polymer moiety and having the structure: 
       
         
           
           
               
               
           
         
       
       wherein
 α is an integer, 1≦α≦7; 
 β is an integer, 1≦β≦7; 
 δ is an integer, 2≦δ≦5; 
 A is either CO, NH, NCO, SO 2 CH 2 CH 2 ; and 
 E is either NH or CO. 
 
     
     
         10 . The compound of  claim 9 , wherein
 α=β=1 or 2;   δ=3;   A is CO; and   E is NH.   
     
     
         11 . A pharmaceutical composition comprising
 (a) a compound according to  claim 9 ;   (b) one or more pharmaceutically acceptable diluents, preservatives, solubilizers, emulsifiers, adjuvants or carriers.   
     
     
         12 . The composition of  claim 11 , wherein
 α=β=1 or 2;   δ=3;   A is CO; and   E is NH.   
     
     
         13 . A compound comprising a peptide moiety, a linker moiety, and a water-soluble polymer moiety, wherein the linker moiety is between the peptide moiety and the water-soluble polymer moiety and having the structure: 
       
         
           
           
               
               
           
         
       
       wherein
 κ is an integer, 0≦κ≦8; 
 ρ is an integer, 0≦κ≦8; 
 τ is an integer, 2≦κ≦5; 
 A is NH or NR; 
 R is alkyl; and 
 E is either NH, CO, or NR, wherein R is H, or alkyl. 
 
     
     
         14 . The compound of  claim 13 , wherein
 κ=ρ=0;   τ=3;   A is either NHR or NR;   R is CH 3 ; and   E is CO.   
     
     
         15 . A pharmaceutical composition comprising:
 (a) a compound according to  claim 13 ;   (b) one or more pharmaceutically acceptable diluents, preservatives, solubilizers, emulsifiers, adjuvants or carriers.   
     
     
         16 . The composition of  claim 15 , wherein
 κ=ρ=0;   τ=3;   A is NR;   R is CH 3 ; and   E is CO.   
     
     
         17 . A compound comprising a peptide moiety, a linker moiety, and a water-soluble polymer moiety, wherein the linker moiety is between the peptide moiety and the water-soluble polymer moiety and having the structure: 
       
         
           
           
               
               
           
         
       
       wherein
 χ is an integer, 1≦χ≦4 
 φ is an integer, 1≦φ≦4; 
 γ is an integer, 2≦γ≦8; 
 η is an integer 2≦η≦8; 
 A is either CO, NH, NCO, or SO 2 CH 2 CH 2 ; 
 B is either CH or N; 
 C is either CO(CH 2 ) σ CO or (CH 2 ) σ ; 
 D is either CH or N; 
 E is either NH or CO; and 
 σ is an integer, 2≦σ≦5. 
 
     
     
         18 . The compound of  claim 17 , wherein
 χ=1;   φ=1;   γ is an integer 2≦γ≦3;   η is an integer 2≦η≦3;   A is CO;   B is N;   C is CO(CH 2 ) σ CO or (CH 2 ) σ ;   D is N;   E is NH; and   σ is an integer, 2≦σ≦3.   
     
     
         19 . A pharmaceutical composition comprising
 (a) a compound according to  claim 17 ;   (b) one or more pharmaceutically acceptable diluents, preservatives, solubilizers, emulsifiers, adjuvants and/or carriers.   
     
     
         20 . The composition of  claim 19 , wherein
 χ=1;   φ=1;   γ is an integer 2≦γ≦3;   η is an integer 2≦η≦3;   A is CO;   B is N;   C is CO(CH 2 ) σ CO or (CH 2 ) σ ;   D is N;   E is NH; and   σ is an integer, 2≦σ≦3.   
     
     
         21 . A compound comprising a peptide moiety, a linker moiety, and a water-soluble polymer moiety, wherein the linker moiety is between the peptide moiety and the water-soluble polymer moiety and having the structure: 
       
         
           
           
               
               
           
         
       
       wherein
 λ is an integer, 1≦λ≦4; 
 μ is an integer, 1≦μ≦4; 
 A is either CO, NH, NCO, or SO 2 CH 2 CH 2 ; 
 B is either CH or N; 
 G is either (CH 2 ) ν , CO or COCH 2 OCH 2 CO; 
 E is either NH or CO; 
 X is either CO, bond, or CONH; 
 Y is either CO, bond, or NHCO; and 
 ν is an integer, 2≦ν≦4. 
 
     
     
         22 . The compound of  claim 21 , wherein
 λ is an integer, 1≦λ≦3;   μ=2;   A is CO;   B is N;   G is either (CH 2 ) ν , CO or COCH 2 OCH 2 CO;   E is NH;   X is CO or bond;   Y is CO or bond; and   ν=2.   
     
     
         23 . A pharmaceutical composition comprising
 (a) a compound according to  claim 21 ; and   (b) one or more pharmaceutically acceptable diluents, preservatives, solubilizers, emulsifiers, adjuvants and/or carriers.   
     
     
         24 . The composition of  claim 23 , wherein
 λ is an integer, 1≦λ≦3;   μ=2;   A is CO;   B is N;   G is either (CH 2 ) ν , CO or COCH 2 OCH 2 CO;   E is NH;   X is CO or bond;   Y is CO or bond; and   ν=2.   
     
     
         25 - 38 . (canceled) 
     
     
         39 . A compound that binds to and activates the erythropoietin receptor (EPO-R), which compound comprises a peptide dimer having the formula: 
       
         
           
           
               
               
           
         
       
       wherein
 (i) in each peptide monomer of the peptide dimer, each amino acid is indicated by standard one letter abbreviation, AcG is N-acetylglycine, 1-Nal is 1-naphthylalanine, and MeG is N-methylglycine; 
 (ii) each peptide monomer of the peptide dimer contains an intramolecular disulfide bond between the two cysteine (C) residues of each monomer; and 
 (iii) PEG comprises two linear polyethylene glycol (PEG) moieties having a combined molecular weight of about 10,000 to about 60,000 Daltons. 
 
     
     
         40 - 41 . (canceled)