Derivatisation of biological molecules

Abstract

The present disclosure relates to a new polymerisation process in which ethylenically unsaturated monomers are polymerised by a living radical polymerisation process in the presence of an initiator and a catalyst. Polymers produced by this new process are also thought to be novel and may be used to derivatise biological molecules to improve their efficacy as therapeutic treatments. A preferred polymer is of formula The polymers are particularly suitable for derivatising proteins, such as interferon-α.

What is claimed is: 
     
         1 . A polymerisation process in which ethylenically unsaturated monomers are polymerised by a living radical polymerisation process in the presence of an initiator of the general formula I or II and a catalyst 
       
         
           
           
               
               
           
         
       
       wherein
 R 1  is an electron-withdrawing group; 
 R 2  is selected from C═O, C(═O)NR 9  or a bond; wherein R 9  is H or C 1-4  alkyl; 
 R 3  is selected from the group consisting of C 1 -C 20  alkylene, C 3 -C 8  cycloalkylene, C(═O)R 10 , C(═O)NR 11 , C 1 -C 20  alkoxy, C 2 -C 20  alkenylene, C 2 -C 20  alkenyl oxiranylene, arylene, heterocyclene and aralkylene; in which 0 to all of the hydrogen atoms are replaced with halogen, C 1 -C 6  alkyl optionally substituted with 1 to 2 substituents selected from the group consisting of C 1 -C 4  alkoxy, aryl, heterocyclyl, C(═O)R 13 , C(═O)NR 11 R 12 , oxiranyl and glycidyl; 
 R 10  is alkylene of from 1 to 20 carbon atoms, alkoxy from 1 to 20 carbon atoms, oligo(alkoxy) in which each alkoxy group has from 1 to 3 carbon atoms, aryloxy or heterocyclyloxy; any of which groups may have substituents selected from optionally substituted alkoxy, oligoalkoxy, amino (including mono- and di-alkyl amino and trialkyl ammonium, which alkyl groups, in turn, may have substituents selected from acyl, alkoxycarbonyl, alkenoxycarbonyl, aryl and hydroxy) and hydroxyl groups; 
 R 11  and R 12  are independently H or alkyl of from 1 to 20 carbon atoms, or R 11  and R 12  may be joined together to form an alkanediyl group of from 2 to 5 carbon atoms, thus forming a 3- to 6-membered ring; 
 R 4  and R 5  are each independently selected from H, Z, halogen, C 1-20  alkyl, C 3 -C 8  cycloalkyl, OH, CN, C 2 -C 20  alkenyl, C 2 -C 20  alkenyl oxiranyl, C(═O)R 13 , glycidyl, aryl, heterocyclyl, arylkyl, aralkenyl, in which 0 to all of the hydrogen atoms are replaced with halogen, C 1 -C 6  alkyl optionally substituted with 1 to 2 substituents selected from the group consisting of C 1 -C 4  alkoxy, aryl, heterocyclyl, C(═O)R 13 , C(═O)NR 11 R 12 , oxiranyl and glycidyl; 
 where R 13  is alkyl of from 1 to 20 carbon atoms, alkoxy of from 1 to 20 carbon atoms, oligo(alkoxy) in which each alkoxy group has 1 to 3 carbon atoms, aryloxy or heterocyclyloxy any of which groups may have substituents selected from optionally substituted alkoxy, oligoalkoxy, amino (including mono- and di-alkyl amino and trialkyl ammonium, which alkyl groups, in turn may have substituents selected from acyl, alkoxycarbonyl, alkenoxycarbonyl, aryl and hydroxy) and hydroxyl groups; and 
 L is a linking group; 
 Z is selected from the group consisting of Cl, Br, I, OR 14 , SR 15 , SeR 15 , OP(═O)R 15 , OP(═O)(═OR 15 ) 2 , O—N(R 15 ) 2  and S—C(═S)N(R 15 ) 2 , where R 14  is alkyl of from 1 to 20 carbon atoms in which each of the hydrogen atoms may be independently replaced by halide, R 15  is aryl or a straight or branched C 1 -C 20  alkyl group, and where an N(R 15 ) 2  group is present, the two R 15  groups may be joined to form a 5- or 6-membered heterocyclic ring; 
 R 6  is CH 2 E 2 , H or C 1-4  alkyl; 
 R 7  and R 8  are each independently selected from H and C 1-4  alkyl; 
 m is 0-4; and 
 E 1  and E 2  are each, independently, a leaving group, or a precursor to a leaving group. 
 
     
     
         2 . A polymerisation process according to  claim 1 , in which R 4  and R 5  are each methyl and R 3  is —CO—R 10  in which R 10  is oligoalkoxy. 
     
     
         3 . A polymerisation process according to  claim 2 , in which R 10  is an oligoethoxy in which there are 2 to 10 ethoxy groups. 
     
     
         4 . A polymerisation process according to  claim 1 , wherein E 1  and E 2  (if present) are each independently selected from —SR 17 , —SO 2 R 17 , —OSO 2 R 17 , —N + R 17   3 , —N + HR 2   17 , —N + H 2 R 17 , halogen, or —OAr, in which R 17  represents an alkyl or aryl group and Ar represents a substituted aryl group containing at least one electron-withdrawing substituent. 
     
     
         5 . A polymerisation process according to  claim 4 , wherein E 1  and E 2  are each, independently, an (optionally substituted) aryl sulfide or aryl sulfone group. 
     
     
         6 . A polymerisation process according to  claim 4 , wherein groups E 1  and E 2  are identical and are a group of formula 
       
         
           
           
               
               
           
         
       
     
     
         7 . A polymerisation process according to  claim 1 , wherein R 1  is selected from a keto group, an ester group, and a sulphone group. 
     
     
         8 . A polymerisation process according to  claim 1 , wherein the linking group, L, is selected from a bond, a C 1-10  alkylene group, or an optionally substituted aryl or heteroaryl, any of which groups may have substituents selected from optionally substituted alkoxy, oligoalkoxy, amino (including mono- and di-alkyl amino and trialkyl ammonium, which alkyl groups, in turn, may have substituents selected from acyl, alkoxycarbonyl, alkenoxycarbonyl, aryl and hydroxy) and hydroxyl groups. 
     
     
         9 . A polymerisation process according to  claim 1 , wherein the initiator of general formula I is 
       
         
           
           
               
               
           
         
       
     
     
         10 . A polymerisation process according to  claim 1 , comprising subsequent reaction step(s) in which groups E 1  and E 2  (if present) which are precursors to leaving groups are converted into leaving groups. 
     
     
         11 . A polymerisation process according to  claim 1 , in which the ethylenically unsaturated monomers are zwitterionic monomers. 
     
     
         12 . A polymerisation process according to  claim 11 , in which the zwitterionic monomers are 2-methacryloyloxyethyl-2′-trimethylammonium ethyl phosphate inner salt. 
     
     
         13 . A compound of general formula II 
       
         
           
           
               
               
           
         
       
       wherein
 R 1  is an electron-withdrawing group; 
 R 2  is selected from C═O, C(═O)NR 9  or a bond; wherein R 9  is H or C 1-4  alkyl; 
 R 3  is selected from the group consisting of C 1 -C 20  alkylene, C 3 -C 8  cycloalkylene, C(═O)R 10 , C(═O)NR 11 , C 1 -C 20  alkoxy, C 2 -C 20  alkenylene, C 2 -C 20  alkenyl oxiranylene, arylene, heterocyclene and aralkylene; in which 0 to all of the hydrogen atoms are replaced with halogen, C 1 -C 6  alkyl optionally substituted with 1 to 2 substituents selected from the group consisting of C 1 -C 4  alkoxy, aryl, heterocyclyl, C(═O)R 13 , C(═O)NR 11 R 12 , oxiranyl and glycidyl; 
 R 10  is alkylene of from 1 to 20 carbon atoms, alkoxy from 1 to 20 carbon atoms, oligo(alkoxy) in which each alkoxy group has from 1 to 3 carbon atoms, aryloxy or heterocyclyloxy; any of which groups may have substituents selected from optionally substituted alkoxy, oligoalkoxy, amino (including mono- and di-alkyl amino and trialkyl ammonium, which alkyl groups, in turn, may have substituents selected from acyl, alkoxycarbonyl, alkenoxycarbonyl, aryl and hydroxy) and hydroxyl groups; 
 R 11  and R 12  are independently H or alkyl of from 1 to 20 carbon atoms, or R 11  and R 12  may be joined together to form an alkanediyl group of from 2 to 5 carbon atoms, thus forming a 3- to 6-membered ring; 
 R 4  and R 5  are each independently selected from H, Z, halogen, C 1-20  alkyl, C 3 -C 8  cycloalkyl, OH, CN, C 2 -C 20  alkenyl, C 2 -C 20  alkenyl oxiranyl, C(═O)R 13 , glycidyl, aryl, heterocyclyl, arylkyl, aralkenyl, in which 0 to all of the hydrogen atoms are replaced with halogen, C 1 -C 6  alkyl optionally substituted with 1 to 2 substituents selected from the group consisting of C 1 -C 4  alkoxy, aryl, heterocyclyl, C(═O)R 13 , C(═O)NR 11 R 12 , oxiranyl and glycidyl; 
 where R 13  is alkyl of from 1 to 20 carbon atoms, alkoxy of from 1 to 20 carbon atoms, oligo(alkoxy) in which each alkoxy group has 1 to 3 carbon atoms, aryloxy or heterocyclyloxy any of which groups may have substituents selected from optionally substituted alkoxy, oligoalkoxy, amino (including mono- and di-alkyl amino and trialkyl ammonium, which alkyl groups, in turn may have substituents selected from acyl, alkoxycarbonyl, alkenoxycarbonyl, aryl and hydroxy) and hydroxyl groups; and 
 L is a linking group; 
 Z is selected from the group consisting of Cl, Br, I, OR 14 , SR 15 , SeR 15 , OP(═O)R 15 , OP(═O)(═OR 15 ) 2 , O—N(R 15 ) 2  and S—C(═S)N(R 15 ) 2 , where R 14  is alkyl of from 1 to 20 carbon atoms in which each of the hydrogen atoms may be independently replaced by halide, R 15  is aryl or a straight or branched C 1 -C 20  alkyl group, and where an N(R 15 ) 2  group is present, the two R 15  groups may be joined to form a 5- or 6-membered heterocyclic ring; 
 R 6  is CH 2 E 2 , H or C 1-4  alkyl; 
 R 7  and R 8  are each independently selected from H and C 1-4  alkyl; and 
 m is 0-4. 
 
     
     
         14 . A compound according to  claim 13 , in which R 4  and R 5  are each methyl and R 3  is —CO—R 10  in which R 10  is oligoalkoxy. 
     
     
         15 . A compound according to  claim 14 , in which R is an oligoethoxy in which there are 2 to 10 ethoxy groups. 
     
     
         16 . A compound according to  claim 13 , wherein R 1  is selected from a keto group, an ester group, and a sulphone group. 
     
     
         17 . A compound according to  claim 13 , wherein the linking group, L, is selected from a bond, a C 1-10  alkylene group, or an optionally substituted aryl or heteroaryl, any of which groups may have substituents selected from optionally substituted alkoxy, oligoalkoxy, amino (including mono- and di-alkyl amino and trialkyl ammonium, which alkyl groups, in turn, may have substituents selected from acyl, alkoxycarbonyl, alkenoxycarbonyl, aryl and hydroxy) and hydroxyl groups. 
     
     
         18 . A polymer of general formula VII or VIII: 
       
         
           
           
               
               
           
         
       
       wherein
 R 1  is an electron-withdrawing group; 
 R 2  is selected from C═O, C(═O)NR 9  or a bond; wherein R 9  is H or C 1-4  alkyl; 
 R 3  is selected from the group consisting of C 1 -C 20  alkylene, C 3 -C 8  cycloalkylene, C(═O)R 10 , C(═O)NR 11 , C 1 -C 20  alkoxy, C 2 -C 20  alkenylene, C 2 -C 20  alkenyl oxiranylene, arylene, heterocyclene and aralkylene; in which 0 to all of the hydrogen atoms are replaced with halogen, C 1 -C 6  alkyl optionally substituted with 1 to 2 substituents selected from the group consisting of C 1 -C 4  alkoxy, aryl, heterocyclyl, C(═O)R 13 , C(═O)NR 11 R 12 , oxiranyl and glycidyl; 
 R 10  is alkylene of from 1 to 20 carbon atoms, alkoxy from 1 to 20 carbon atoms, oligo(alkoxy) in which each alkoxy group has from 1 to 3 carbon atoms, aryloxy or heterocyclyloxy; any of which groups may have substituents selected from optionally substituted alkoxy, oligoalkoxy, amino (including mono- and di-alkyl amino and trialkyl ammonium, which alkyl groups, in turn, may have substituents selected from acyl, alkoxycarbonyl, alkenoxycarbonyl, aryl and hydroxy) and hydroxyl groups; 
 R 11  and R 12  are independently H or alkyl of from 1 to 20 carbon atoms, or R 11  and R 12  may be joined together to form an alkanediyl group of from 2 to 5 carbon atoms, thus forming a 3- to 6-membered ring; 
 R 4  and R 5  are each independently selected from H, Z, halogen, C 1-20  alkyl, C 3 -C 8  cycloalkyl, OH, CN, C 2 -C 20  alkenyl, C 2 -C 20  alkenyl oxiranyl, C(═O)R 13 , glycidyl, aryl, heterocyclyl, arylkyl, aralkenyl, in which 0 to all of the hydrogen atoms are replaced with halogen, C 1 -C 6  alkyl optionally substituted with 1 to 2 substituents selected from the group consisting of C 1 -C 4  alkoxy, aryl, heterocyclyl, C(═O)R 13 , C(═O)NR 11 R 12 , oxiranyl and glycidyl; 
 where R 13  is alkyl of from 1 to 20 carbon atoms, alkoxy of from 1 to 20 carbon atoms, oligo(alkoxy) in which each alkoxy group has 1 to 3 carbon atoms, aryloxy or heterocyclyloxy any of which groups may have substituents selected from optionally substituted alkoxy, oligoalkoxy, amino (including mono- and di-alkyl amino and trialkyl ammonium, which alkyl groups, in turn may have substituents selected from acyl, alkoxycarbonyl, alkenoxycarbonyl, aryl and hydroxy) and hydroxyl groups; and 
 L is a linking group; 
 Z is selected from the group consisting of Cl, Br, I, OR 14 , SR 15 , SeR 15 , OP(═O)R 15 , OP(═O)(═OR 15 ) 2 , O—N(R 15 ) 2  and S—C(═S)N(R 15 ) 2 , where R 14  is alkyl of from 1 to 20 carbon atoms in which each of the hydrogen atoms may be independently replaced by halide, R 15  is aryl or a straight or branched C 1 -C 20  alkyl group, and where an N(R 15 ) 2  group is present, the two R 15  groups may be joined to form a 5- or 6-membered heterocyclic ring; 
 R 6  is CH 2 E 2 , H or C 1-4  alkyl; 
 R 7  and R 8  are each independently selected from H and C 1-4  alkyl; 
 m is 0-4; 
 E 1  and E 2  are each, independently, a leaving group, or a precursor to a leaving group; 
 f is 1 to 500; and 
 Groups M are the same or different and are residues of ethylenically unsaturated monomers. 
 
     
     
         19 . A polymer according to  claim 18 , in which R 4  and R 5  are each methyl and R 3  is —CO—R 10  in which R 10  is oligoalkoxy. 
     
     
         20 . A polymer according to  claim 19 , in which R 10  is an oligoethoxy in which there are 2 to 10 ethoxy groups. 
     
     
         21 . A polymer according to  claim 18 , wherein E 1  and E 2  (if present) are each independently selected from —SR 17 , —SO 2 R 17 , —OSO 2 R 17 , —N + R 17   3 , —N + HR 2   17 , —N + H 2 R 17 , halogen, or —OAr, in which R 17  represents an alkyl or aryl group and Ar represents a substituted aryl group containing at least one electron-withdrawing substituent. 
     
     
         22 . A polymer according to  claim 21 , wherein E 1  and E 2  are each, independently, an (optionally substituted) aryl sulfide or aryl sulfone group. 
     
     
         23 . A polymer according to  claim 21 , wherein groups E 1  and E 2  are identical and are a group of formula 
       
         
           
           
               
               
           
         
       
     
     
         24 . A polymer according to  claim 18 , wherein R 1  is selected from a keto group, an ester group, and a sulphone group. 
     
     
         25 . A polymer according to  claim 18 , wherein the linking group, L, is selected from a bond, a C 1-10  alkylene group, or an optionally substituted aryl or heteroaryl, any of which groups may have substituents selected from optionally substituted alkoxy, oligoalkoxy, amino (including mono- and di-alkyl amino and trialkyl ammonium, which alkyl groups, in turn, may have substituents selected from acyl, alkoxycarbonyl, alkenoxycarbonyl, aryl and hydroxy) and hydroxyl groups. 
     
     
         26 . A polymer according to  claim 18 , in which residues M are selected from radicals of the general formula IX 
       
         
           
           
               
               
           
         
       
       in which R 18  is selected from hydrogen, halogen, C 1-4  alkyl and groups COOR 22  in which R 22  is hydrogen and C 1-4  alkyl;
 R 19  is selected from hydrogen, halogen and C 1-4  alkyl; 
 R 20  is selected from hydrogen, halogen, C 1-4  alkyl and groups COOR 22  provided that R 18  and R 20  are not both COOR 22 ; and 
 R 21  is a C 1-10  alkyl, a C 1-20  alkoxycarbonyl, a mono- or di-(C 1-20  alkyl) amino carbonyl, a C 6-20  aryl (including alkaryl), a C 7-20  aralkyl, a C 6-20  aryloxycarbonyl, a C 1-20  aralkyloxycarbonyl, a C 6-20  arylamino carbonyl, a C 7-20  aralkyl-amino, a hydroxyl or a C 2-10  acyloxy group, any of which may have one or more substituents selected from halogen atoms, alkoxy, oligo-alkoxy, aryloxy, acyloxy, acylamino, amine (including mono and di-alkyl amino and trialkylammonium), carboxyl, sulphonyl, phosphoryl, phosphino, (including mono- and di-alkyl phosphine and tri-alkylphosphonium), zwitterionic and hydroxyl groups; 
 or R 21  and R 20  or R 21  and R 19  may together form —CONR 23 CO in which R 23  is a C 1-20  alkyl group. 
 
     
     
         27 . A polymer according to  claim 18  of formula 
       
         
           
           
               
               
           
         
       
       wherein f is 1 to 500. 
     
     
         28 . Method of derivatisation of a biological molecule in which a polymer of general formula X or general formula XI 
       
         
           
           
               
               
           
         
       
       is added to a biological molecule of general formula T 1 D 1 X or to biological molecules of general formula T 1 D 1 X and T 2 D 2 X, wherein D is S, NH or O and X is H, D 1 T 1  or D 2 T 2  to form an adduct of general formula XII or of general formula XIII 
       
         
           
           
               
               
           
         
       
       wherein Groups M are the same or different and are residues of ethylenically unsaturated monomers;
 f is 1 to 500; 
 R 36  is —CH 2 E 4 , H or C 1-4  alkyl; and 
 E 3  and E 4  are each, independently, a leaving group, 
 x is 1 or 2; 
 p is 1-5; 
 q is 1-10; 
 R 1  is an electron-withdrawing group; 
 R 2  is selected from C═O, C(═O)NR 9  or a bond; wherein R 9  is H or C 1-4  alkyl; 
 R 3  is selected from the group consisting of C 1 -C 20  alkylene, C 3 -C 8  cycloalkylene, C(═O)R 10 , C(═O)NR 11 , C 1 -C 20  alkoxy, C 2 -C 20  alkenylene, C 2 -C 20  alkenyl oxiranylene, arylene, heterocyclene and aralkylene; in which 0 to all of the hydrogen atoms are replaced with halogen, C 1 -C 6  alkyl optionally substituted with 1 to 2 substituents selected from the group consisting of C 1 -C 4  alkoxy, aryl, heterocyclyl, C(═O)R 13 , C(═O)NR 11 R 12 , oxiranyl and glycidyl; 
 R 10  is alkylene of from 1 to 20 carbon atoms, alkoxy from 1 to 20 carbon atoms, oligo(alkoxy) in which each alkoxy group has from 1 to 3 carbon atoms, aryloxy or heterocyclyloxy; any of which groups may have substituents selected from optionally substituted alkoxy, oligoalkoxy, amino (including mono- and di-alkyl amino and trialkyl ammonium, which alkyl groups, in turn, may have substituents selected from acyl, alkoxycarbonyl, alkenoxycarbonyl, aryl and hydroxy) and hydroxyl groups; 
 R 11  and R 12  are independently H or alkyl of from 1 to 20 carbon atoms, or R 11  and R 12  may be joined together to form an alkanediyl group of from 2 to 5 carbon atoms, thus forming a 3- to 6-membered ring; 
 R 4  and R 5  are each independently selected from H, Z, halogen, C 1-20  alkyl, C 3 -C 8  cycloalkyl, OH, CN, C 2 -C 20  alkenyl, C 2 -C 20  alkenyl oxiranyl, C(═O)R 13 , glycidyl, aryl, heterocyclyl, arylkyl, aralkenyl, in which 0 to all of the hydrogen atoms are replaced with halogen, C 1 -C 6  alkyl optionally substituted with 1 to 2 substituents selected from the group consisting of C 1 -C 4  alkoxy, aryl, heterocyclyl, C(═O)R 13 , C(═O)NR 11 R 12 , oxiranyl and glycidyl; 
 where R 13  is alkyl of from 1 to 20 carbon atoms, alkoxy of from 1 to 20 carbon atoms, oligo(alkoxy) in which each alkoxy group has 1 to 3 carbon atoms, aryloxy or heterocyclyloxy any of which groups may have substituents selected from optionally substituted alkoxy, oligoalkoxy, amino (including mono- and di-alkyl amino and trialkyl ammonium, which alkyl groups, in turn may have substituents selected from acyl, alkoxycarbonyl, alkenoxycarbonyl, aryl and hydroxy) and hydroxyl groups; and 
 L is a linking group; 
 Z is selected from the group consisting of Cl, Br, I, OR 14 , SR 15 , SeR 15 , OP(═O)R 15 , OP(═O)(═OR 15 ) 2 , O—N(R 15 ) 2  and S—C(═S)N(R 15 ) 2 , where R 14  is alkyl of from 1 to 20 carbon atoms in which each of the hydrogen atoms may be independently replaced by halide, R 15  is aryl or a straight or branched C 1 -C 20  alkyl group, and where an N(R 15 ) 2  group is present, the two R 15  groups may be joined to form a 5- or 6-membered heterocyclic ring; 
 R 7  and R 8  are each independently selected from H and C 1-4  alkyl; and 
 m is 0-4. 
 
     
     
         29 . A method according to  claim 28 , in which R 4  and R 5  are each methyl and R 3  is —CO—R 10  in which R 10  is oligoalkoxy. 
     
     
         30 . A method according to  claim 29 , in which R is an oligoethoxy in which there are 2 to 10 ethoxy groups. 
     
     
         31 . A method according to  claim 28 , wherein R 1  is selected from a keto group, an ester group, and a sulphone group. 
     
     
         32 . A method according to  claim 28 , wherein the linking group, L, is selected from a bond, a C 1-10  alkylene group, or an optionally substituted aryl or heteroaryl, any of which groups may have substituents selected from optionally substituted alkoxy, oligoalkoxy, amino (including mono- and di-alkyl amino and trialkyl ammonium, which alkyl groups, in turn, may have substituents selected from acyl, alkoxycarbonyl, alkenoxycarbonyl, aryl and hydroxy) and hydroxyl groups. 
     
     
         33 . A method according to  claim 28 , wherein the biological molecules is/are a peptide, protein or a lipoprotein. 
     
     
         34 . A method according to  claim 28 , wherein the group or groups D 1 X/D 2 X are thiol groups, SH. 
     
     
         35 . A method according to  claim 34 , wherein the group or groups D 1 X/D 2 X are part of terminal cysteine residues on the protein(s), lipoprotein(s) or peptide(s).