Methods for preparing polymeric reagents and compositions of polymeric reagents

Abstract

Methods for preparing active carbonate esters of water-soluble polymers are provided. Also provided are other methods related to the active carbonate esters of water-soluble polymers, as well as corresponding compositions.

What is claimed is: 
     
         1 . A synthetic method comprising:
 (a) combining a composition comprising an aprotic solvent and an amine-terminated or hydroxy-terminated, water-soluble polymer with a composition comprising an activated carbonate reagent, optionally in the presence of a catalyst or acid-neutralizing base, wherein the composition comprising the activated carbonate reagent is added such that there is an excess of the activated carbonate reagent relative to the amine-terminated or hydroxy-terminated, water-soluble polymer, to thereby result in a composition comprising an active carbonate ester of the water-soluble polymer and unreacted activated carbonate reagent; and   (b) adding a composition comprising a reactive molecule to the composition comprising the active carbonate ester of the water-soluble polymer and unreacted activated carbonate reagent, wherein the composition comprising a reactive molecule is added such that substantially all of the unreacted activated carbonate reagent is substantially consumed.   
     
     
         2 . The method of  claim 1 , wherein the active carbonate ester of the water-soluble polymer has a structure of the following formula:
   POLY-O—(C═O)—O—R
   
       wherein:
 POLY is a water-soluble polymer; and 
 R is an organic radical, 
 with the proviso that when POLY is a linear, methyl-capped poly(ethylene glycol), the linear, methyl-capped polyethylene glycol has a molecular weight of at least 103. 
 
     
     
         3 . The method as in any one of  claims 1  and  2 , wherein the activated carbonate reagent has a structure of one of the following formulae: R—O(C═O)—OR; R—O(C═O)—OR′; and R—O(C═O)X, wherein R is an organic radical, R′ is an organic radical different than R, and X is Cl, Br, or I. 
     
     
         4 . The method as in any one of  claims 1 ,  2  and  3 , wherein the aprotic solvent is selected from the group consisting of acetonitrile, dimethylsulfoxide, dimethylformamide, chloroform, dichloromethane, and combinations thereof. 
     
     
         5 . The method of  claim 1 , further comprising reacting the active carbonate ester of the water-soluble polymer in one or more reactions to form a polymeric reagent. 
     
     
         6 . The method of  claim 1 , wherein the active carbonate ester of the water-soluble polymer is isolated by distilling off the solvent. 
     
     
         7 . The method of  claim 1 , wherein the active carbonate ester of the water-soluble polymer is isolated by precipitation upon addition of a non-solvent. 
     
     
         8 . The method of  claim 7 , wherein the non-solvent is selected from the group consisting of isopropyl alcohol, hexane, ethyl ether, and combinations thereof. 
     
     
         9 . The method of  claim 1 , wherein the amine-terminated or hydroxy-terminated, water-soluble polymer is a water-soluble polymer bearing a terminal amine group or hydroxy group, wherein the water-soluble polymer is selected from the group consisting of poly(alkylene glycol), poly(olefinic alcohol), poly(vinylpyrrolidone), poly(hydroxyalkylmethacrylamide), poly(hydroxyalkylmethacrylate), poly(saccharide), poly(α-hydroxyacetic acid), poly(acrylic acid), poly(vinyl alcohol), polyphosphazene, polyoxazoline, poly(N-acryloylmorpholine), and copolymers or terpolymers thereof. 
     
     
         10 . The method of  claim 6 , wherein the water-soluble polymer is a poly(ethylene glycol). 
     
     
         11 . The method of  claim 10 , wherein the poly(ethylene glycol) is a linear poly(ethylene glycol). 
     
     
         12 . The method of  claim 10 , wherein the poly(ethylene glycol) is a branched poly(ethylene glycol). 
     
     
         13 . The method of  claim 10 , wherein the poly(ethylene glycol) is a multi-arm poly(ethylene glycol). 
     
     
         14 . The method of  claim 10 , wherein the poly(ethylene glycol) has a molecular weight of about 100 to about 100,000 Daltons. 
     
     
         15 . The method as in any one of  claims 1 ,  2 ,  3  and  4 , wherein the amine-terminated or hydroxy-terminated, water-soluble polymer is an amine-terminated, water-soluble polymer. 
     
     
         16 . The method as in any one of  claims 1 ,  2 ,  3  and  4 , wherein the amine-terminated or hydroxy-terminated, water-soluble polymer is a hydroxy-terminated, water-soluble polymer. 
     
     
         17 . The method as in any one of  claims 1 ,  2 ,  3  and  4 , wherein the activated carbonate reagent is selected from the group consisting of di(1-benzotriazolyl) carbonate, disuccinimidyl carbonate, p-nitrophenyl chloroformate, trichlorophenyl chloroformate, p-nitrophenyl succinimidyl carbonate, p-nitrophenyl 1-benzotriazolyl carbonate, pentafluorophenyl chloroformate, and 1,1′-carbonyldiimidazole. 
     
     
         18 . The method as in any one of  claims 1 ,  2 ,  3  and  4 , wherein the excess of the activated carbonate reagent relative to the amine-terminated or hydroxy-terminated, water-soluble polymer represents at least about a 5 mol % excess. 
     
     
         19 . The method as in any one of  claims 1 ,  2 ,  3  and  4 , wherein the excess of the activated carbonate reagent relative to the amine-terminated or hydroxy-terminated, water-soluble polymer represents at least about a 50 mol % excess. 
     
     
         20 . The method as in any one of  claims 1 ,  2 ,  3  and  4 , wherein the excess of the activated carbonate reagent relative to the amine-terminated or hydroxy-terminated, water-soluble polymer represents at least about a two fold molar excess. 
     
     
         21 . The method as in any one of  claims 1 ,  2 ,  3  and  4 , wherein the reactive molecule is selected from the group consisting of water, a lower alkyl monohydric alcohol, hydrogen sulfide, and a lower alkyl monobasic amine. 
     
     
         22 . The method as in  claim 21 , wherein the reactive molecule is water. 
     
     
         23 . The method as in any one of  claims 1 ,  2 ,  3  and  4 , wherein the reactive molecule is selected from hydroxyalkyl attached to a solid support, mercaptoalkyl attached to a solid support, primary amine attached to a solid support, and secondary amine attached to a secondary support. 
     
     
         24 . A synthetic method comprising:
 (a) combining a composition comprising an aprotic solvent and a hydroxy-terminated, water-soluble polymer having the following structure:
   CH 3 O—(CH 2 CH 2 O) n —CH 2 CH 2 —OH
 
   
       wherein (n) is an integer from 2 to about 4000, with a composition comprising di(1-benzotriazolyl) carbonate, wherein the composition comprising the di(1-benzotriazolyl) carbonate is added such that there is an excess of the di(1-benzotriazolyl) carbonate relative to the hydroxy-terminated, water-soluble polymer, to thereby result in a composition comprising an active carbonate ester of the water-soluble polymer having the following structure: 
       
         
           
           
               
               
           
         
       
       wherein (n) is an integer from 2 to about 4000, and unreacted di(1-benzotriazolyl) carbonate;
 (b) adding a composition comprising a reactive molecule to the composition comprising the active carbonate ester of a water-soluble polymer and unreacted di(1-benzotriazolyl) carbonate, wherein the composition comprising the reactive compound is added such that substantially all of the unreacted di(1-benzotriazolyl) carbonate is substantially consumed. 
 
     
     
         25 . The method as in  claim 24 , wherein the reactive molecule is selected from the group consisting of water, a lower alkyl monohydric alcohol, hydrogen sulfide, and a lower alkyl monobasic amine. 
     
     
         26 . The method as in  claim 24 , wherein the reactive molecule is selected from hydroxyalkyl attached to a solid support, mercaptoalkyl attached to a solid support, primary amine attached to a solid support, and secondary amine attached to a secondary support. 
     
     
         27 . A synthetic method comprising:
 (a) combining a composition comprising an aprotic solvent and a hydroxy-terminated, water-soluble polymer having the following structure:
   CH 3 O—(CH 2 CH 2 O) n —CH 2 CH 2 —OH
 
   
       wherein (n) is an integer from 2 to about 4000, with a composition comprising disuccinimidyl carbonate, wherein the composition comprising the disuccinimidyl carbonate is added such that there is an excess of the disuccinimidyl carbonate relative to the hydroxy-terminated, water-soluble polymer, to thereby result in a composition comprising an active carbonate ester of the water-soluble polymer having the following structure: 
       
         
           
           
               
               
           
         
       
       wherein (n) is an integer from 2 to about 4000, and unreacted disuccinimidyl carbonate;
 (b) adding a composition comprising a reactive molecule to the composition comprising the active carbonate ester of the water-soluble polymer and unreacted disuccinimidyl carbonate, wherein the composition comprising the reactive molecule is added such that substantially all of the unreacted disuccinimidyl carbonate is substantially consumed. 
 
     
     
         28 . The method as in  claim 27 , wherein the reactive molecule is selected from the group consisting of water, a lower alkyl monohydric alcohol, hydrogen sulfide, and a lower alkyl monobasic amine. 
     
     
         29 . The method as in  claim 27 , wherein the reactive molecule is selected from hydroxyalkyl attached to a solid support, mercaptoalkyl attached to a solid support, primary amine attached to a solid support, and secondary amine attached to a secondary support. 
     
     
         30 . A synthetic method comprising:
 (a) combining a composition comprising an aprotic solvent and a hydroxy-terminated, water-soluble polymer having the following structure:
   CH 3 O—(CH 2 CH 2 O) n —CH 2 CH 2 —OH
 
   
       wherein (n) is an integer from 2 to about 4000, with a composition comprising p-nitrophenyl chloroformate, wherein the composition comprising the p-nitrophenyl chloroformate is added such that there is an excess of the p-nitrophenyl chloroformate relative to the hydroxy-terminated, water-soluble polymer, to thereby result in a composition comprising an active carbonate ester of the water-soluble polymer having the following structure: 
       
         
           
           
               
               
           
         
       
       wherein (n) is an integer from 2 to about 4000, and unreacted p-nitrophenyl chloroformate;
 (b) adding a composition comprising a reactive molecule to the composition comprising the active carbonate ester of water-soluble polymer and unreacted p-nitrophenyl chloroformate, wherein the composition comprising the reactive molecule is added such that substantially all of the unreacted p-nitrophenyl chloroformate is substantially consumed. 
 
     
     
         31 . The method as in  claim 30 , wherein the reactive molecule is selected from the group consisting of water, a lower alkyl monohydric alcohol, hydrogen sulfide, and a lower alkyl monobasic amine. 
     
     
         32 . The method as in  claim 30 , wherein the reactive molecule is selected from hydroxyalkyl attached to a solid support, mercaptoalkyl attached to a solid support, primary amine attached to a solid support and secondary amine attached to a secondary support. 
     
     
         33 . The composition resulting from carrying out the method of any of the methods of  claims 1  to  32 . 
     
     
         34 . A synthetic method comprising
 reacting an active carbonate ester of a water-soluble polymer prepared by
 (i) combining a composition comprising an aprotic solvent and an amine-terminated or hydroxy-terminated, water-soluble polymer with a composition comprising an activated carbonate reagent, optionally in the presence of a catalyst or acid-neutralizing base, wherein the composition comprising the activated carbonate reagent is added such that there is an excess of the activated carbonate reagent relative to the amine-terminated or hydroxy-terminated, water-soluble polymer, to thereby result in a composition comprising an active carbonate ester of the water-soluble polymer and unreacted activated carbonate reagent; and 
 (ii) adding a composition comprising a reactive molecule to the composition comprising the active carbonate ester of the water-soluble polymer and unreacted activated carbonate reagent, wherein the composition comprising a reactive molecule is added such that substantially all of the unreacted activated carbonate reagent is substantially consumed, 
   with an active agent under conjugation conditions to thereby result in a water-soluble polymer-active agent conjugate.   
     
     
         35 . A synthetic method comprising:
 reacting a polymeric reagent prepared by
 (a) combining a composition comprising an aprotic solvent and an amine-terminated or hydroxy-terminated, water-soluble polymer with a composition comprising an activated carbonate reagent, optionally in the presence of a catalyst or acid-neutralizing base, wherein the composition comprising the activated carbonate reagent is added such that there is an excess of the activated carbonate reagent relative to the amine-terminated or hydroxy-terminated, water-soluble polymer, to thereby result in a composition comprising an active carbonate ester of the water-soluble polymer and unreacted activated carbonate reagent; 
 (b) adding a composition comprising a reactive molecule to the composition comprising the active carbonate ester of the water-soluble polymer and unreacted activated carbonate reagent, wherein the composition comprising a reactive molecule is added such that substantially all of the unreacted activated carbonate reagent is substantially consumed; and 
 (c) reacting the active carbonate ester of the water-soluble polymer in one or more reactions to form a polymeric reagent 
   with an active agent under conjugation conditions to thereby result in a water-soluble polymer-active agent conjugate.