Thiol-Selective Water-Soluble Polymer Derivatives

Abstract

The present invention provides water-soluble, polymer derivatives having a thiol-selective terminus suitable for selective coupling to thiol groups, such as those contained in the cysteine residues of proteins, as well as methods for preparing the water-soluble, polymer derivatives having a thiol-selective terminus.

1 . A method for preparing a thiol-selective derivative of a water-soluble polymer, said method comprising the steps of:
 (i) providing an electrophilically-activated polymer, POLY-L 0,1 -E,   wherein:
 POLY is a water-soluble polymer segment, 
 L is an optional linker, and 
 E is an electrophile selected from the group consisting of carboxylic acid and an activated carboxylic acid derivative, and 
   (ii) reacting said POLY-L 0,1 -E with NU-Y—S, wherein NU-Y—S is N-(2-amino-ethyl)-3-maleimido-propionamide,   
       to thereby form 
       
         
           
           
               
               
           
         
       
     
     
         2 . The method of  claim 1 , wherein L 1  is present and is a linker selected from the group consisting of C 1 -C 10  alkyl and C 1 -C 10  substituted alkyl. 
     
     
         3 . The method of  claim 2 , wherein L 1 is present and is a linker selected from the group consisting of (CH 2 ) 1, 2, 3, 4 and 5 . 
     
     
         4 . The method of  claim 3 , wherein E is an activated carboxylic acid derivative. 
     
     
         5 . The method of  claim 4 , wherein N-hydroxysuccinimidyl ester. 
     
     
         6 . The method of  claim 1 , wherein POLY is a polyalkylene oxide. 
     
     
         7 . The method of  claim 1 , wherein POLY is a polyethylene glycol having the structure Z—(CH 2 CH 2 O) n CH 2 CH 2 —, wherein n ranges from 10 to about 4,000, and Z is or includes a functional group or an end-capping group. 
     
     
         8 . The method of  claim 7 , wherein Z is selected from the group consisting of hydroxy, amino, ester, carbonate, aldehyde, acetal, aldehyde hydrate, ketone, ketal, ketone hydrate, alkenyl, acrylate, methacrylate, acrylamide, sulfone, thiol, carboxylic acid, isocyanate, isothiocyanate, hydrazide, urea, maleimide, vinylsulfone, dithiopyridine, vinylpyridine, iodoacetamide, alkoxy, benzyloxy, silane, lipid, phospholipid, biotin, and fluorescein, including activated and protected forms thereof where applicable. 
     
     
         9 . The method of  claim 7 , wherein Z is selected from the group consisting of N-hydroxysuccinimidyl ester, 1-hydroxybenzotriazolyl carbonate, amine, vinylsulfone, maleimide, N-succinimidyl carbonate, hydrazide, succinimidyl propionate, succinimidyl butanoate, succinimidyl succinate, succinimidyl ester, glycidyl ether, oxycarbonylimidazole, p-nitrophenyl carbonate, aldehyde, orthopyridyl-disulfide, and acrylol. 
     
     
         10 . A thiol-selective derivative of a water-soluble polymer, encompassed by the following structure: 
       
         
           
           
               
               
           
         
         wherein: 
         POLY is a water-soluble polymer segment; and 
         L is an optional linker. 
       
     
     
         11 . The method of  claim 10 , wherein L 1  is present and is a linker selected from the group consisting of C 1 -C 10  alkyl and C 1 -C 10  substituted alkyl. 
     
     
         12 . The method of  claim 11 , wherein L 1  is present and is a linker selected from the group consisting of (CH 2 ) 1, 2, 3, 4 and 5 . 
     
     
         13 . The method of  claim 10 , wherein POLY is a polyalkylene oxide. 
     
     
         14 . The method of  claim 10 , wherein POLY is a polyethylene glycol having the structure Z—(CH 2 CH2O) n —CH 2 CH 2 —, wherein n ranges from 10 to about 4,000, and Z is or includes a functional group or an end-capping group. 
     
     
         15 . The method of  claim 14 , wherein Z is selected from the group consisting of hydroxy, amino, ester, carbonate, aldehyde, acetal, aldehyde hydrate, ketone, ketal, ketone hydrate, alkenyl, acrylate, methacrylate, acrylamide, sulfone, thiol, carboxylic acid, isocyanate, isothiocyanate, hydrazide, urea, maleimide, vinylsulfone, dithiopyridine, vinylpyridine, iodoacetamide, alkoxy, benzyloxy, silane, lipid, phospholipid, biotin, and fluorescein, including activated and protected forms thereof where applicable. 
     
     
         16 . The method of  claim 14 , wherein Z is selected from the group consisting of N-hydroxysuccinimidyl ester, 1-hydroxybenzotriazolyl carbonate, amine, vinylsulfone, maleimide, N-succinimidyl carbonate, hydrazide, succinimidyl propionate, succinimidyl butanoate, succinimidyl succinate, succinimidyl ester, glycidyl ether, oxycarbonylimidazole, p-nitrophenyl carbonate, aldehyde, orthopyridyl-disulfide, and acrylol.