Releasable polymeric conjugates based on aliphatic biodegradable linkers

Abstract

Activated polymeric bicine derivatives such as Formula (I), as well as conjugates made therewith are disclosed. Methods of making and using the bicine derivatives are also disclosed.

1 . A compound comprising the Formula (I):  
       
         
           
           
               
               
           
         
         wherein:  
         R 1  and R 2  are independently selected from the group consisting of substantially non-antigenic polymer residues, H, C 1-6  alkyls, C 2-6  alkenyl, C 2-6  alkynyl, aralkyls, and terminal branching groups;  
         Z is selected from among moieties actively transported into a target cell, hydrophobic moieties, bifunctional linking moieties and combinations thereof,  
         Y 1-3  may be the same or different and are selected from among O, S or NR 11 ;  
         L 1  and L 2  may be the same or different bifunctional linkers;  
         R 3 -R 11 , R 24  and R 25  may be the same or different and are selected from the group consisting of hydrogen, C 1-6 alkyls, C 2-6  alkenyl, C 2-6  alkynyl, C 3-19  branched alkyls, C 3-8  cycloalkyls, C 1-6  substituted alkyls, C 2-6  substituted alkenyls, C 2-6  substituted alkynyls, C 3-8  substituted cycloalkyls, aryls, substituted aryls, aralkyls, C 1-6  heteroalkyls, substituted C 1-6  heteroalkyls, C 1-6  alkoxy, phenoxy and C 1-6  heteroalkoxy,  
         L 3  and L 4  may be the same or different and are selected from:  
           —C(O)(CR 30 R 31 )Y 15 (CR 32 R 33 )C(O)— or  —C(O)(CR 30 R 31 )(CR 32 R 33 )C(O)— wherein: 
 Y 15  is selected from O, S, NR 34  or CH 2 , and  
 R 30-34  may be the same or different and are selected from H, alkyl, alkenyl, alkynyl, heteroalkyl or aryl;  
   
         A is selected from among leaving groups, functional groups, biologically active moieties and OH;  
         a, b, c, d, and e are independently 0 or 1  
         m, n, o, and p are independently positive integers,  
         f and g are 0 or 1, provided that  
         at least one of (f+a) or (g+c) is equal to 2.  
       
     
     
         2 . The compound of  claim 1 , wherein R 3 -R 10 , R 24-25  and R 30-34  are each hydrogen; and Y 15  is O or NR 34 .  
     
     
         3 . The compound of  claim 1 , wherein a, b, c, d, f, g, m, n, o and p are each 1, and e is 0 or 1.  
     
     
         4 . The compound of  claim 1 , wherein (c and g) are each 0.  
     
     
         5 . The compound of  claim 1 , wherein (a and f) are each 0.  
     
     
         6 . The compound of  claim 1 , wherein (c, g, and d) are each 0.  
     
     
         7 . The compound of  claim 1 , wherein (a, b and f) are each 0.  
     
     
         8 . The compound of  claim 1 , wherein R 1  comprises a polyalkylene oxide.  
     
     
         9 . The compound of  claim 1 , wherein R 2  comprises a polyalkylene oxide.  
     
     
         10 . The compound of  claim 1 , wherein R 1  comprises a polyethylene glycol.  
     
     
         11 . The compound of  claim 1  wherein R 2  comprises a polyethylene glycol.  
     
     
         12 . The compound of  claim 1  wherein R 1  or R 2  further include a capping group J, selected from the group consisting of OH, NH 2 , SH, CO 2 H, C 1-6  alkyl moieties,  
       
         
           
           
               
               
           
         
       
     
     
         13 . A compound of  claim 10 , selected from the group consisting of:  
       
         
           
           
               
               
           
         
       
     
     
         14 . The compound of  claim 1 , wherein R 1  is selected from the group consisting of:  
         J-O—(CH 2 CH 2 O) x — J-O—(CH 2 CH 2 O) x —CH 2 C(O)—O—,  J-O—(CH 2 CH 2 O) x —CH 2 CH 2 NR 12 —,  J-O—(CH 2 CH 2 O) x —CH 2 CH 2 SH—,  —OC(O)CH 2 —O—(CH 2 CH 2 O) x —CH 2 C(O)—O—,  —NR 12 CH 2 CH 2 —O—(CH 2 CH 2 O) x —CH 2 CH 2 NR 12 — and  —SHCH 2 CH 2 —O—(CH 2 CH 2 O) x —CH 2 CH 2 SH—.  wherein: 
 x is the degree of polymerization;  
 R 12  is selected from the group consisting of hydrogen, C 1-6  alkyls, C 2-6  alkenyl, C 2-6  alkynyl, C 3-19  branched alkyls, C 3-8  cycloalkyls, C 1-6  substituted alkyls, C 2-6  substituted alkenyls, C 2-6  substituted alkynyls, C 3-8  substituted cycloalkyls, aryls, substituted aryls, aralkyls, C 1-6  heteroalkyls, substituted C 1-6  heteroalkyls, C 1-6  alkoxy, phenoxy and C 1-6  heteroalkoxy,  
 and J is a capping group.  
   
     
     
         15 . The compound of  claim 1 , wherein R 2  is selected from group consisting of:  
         J-O—(CH 2 CH 2 O) x — J-O—(CH 2 CH 2 O) x —CH 2 C(O)—O—,  J-O—(CH 2 CH 2 O) x —CH 2 CH 2 NR 13 —,  J-O—(CH 2 CH 2 O) x —CH 2 CH 2 SH—,  —OC(O)CH 2 —O—(CH 2 CH 2 O) x —CH 2 C(O)—O—,  —NR 13 CH 2 CH 2 —O—(CH 2 CH 2 O) x —CH 2 CH 2 NR 13 — and  —SHCH 2 CH 2 —O—(CH 2 CH 2 O) x —CH 2 CH 2 SH—,  wherein: 
 x is the degree of polymerization;  
 R 13  is selected from the group consisting of hydrogen, C 1-6  alkyls, C 2-6  alkenyls, C 2-6  alkynyls, C 3-12  branched alkyls, C 3-8  cycloalkyls, C 1-6  substituted alkyls, C 2-6  substituted alkenyls, C 2-6  substituted alkynyls, C 3-8  substituted cycloalkyls, aryls substituted aryls, aralkyls, C 1-6  heteroalkyls, substituted C 1-6  heteroalkyls, C 1-6 alkoxy, phenoxy and C 1-6  heteroalkoxy, and J is a capping group.  
   
     
     
         16 . The compound of  claim 1 , wherein R 1-2  are individually selected from the group consisting of:  
         CH 3 —O—(CH 2 CH 2 O) x —,  CH 3 —O—(CH 2 CH 2 O) x —CH 2 C(O)—O—,  CH 3 —O—(CH 2 CH 2 O) x —C 2 CH 2 NH— and  CH 3 —O—(CH 2 CH 2 O) x —CH 2 CH 2 SH— wherein x is the degree of polymerization.    
     
     
         17 . The compound of  claim 1 , wherein R 1  and R 2  each comprise a polymer residue of the formula  
       
         
           
           
               
               
           
         
         wherein x is the degree of polymerization.  
       
     
     
         18 . The compound of  claim 17 , wherein R 1  and R 2  each have a weight average molecular weight of from about 2,000 Da to about 25,000 Da.  
     
     
         19 . The compound of  claim 1  wherein L 1  and L 2  are independently selected from the group consisting of:  
       
         
           
           
               
               
           
         
         wherein: 
 R 14 -R 17  and R 19  are independently selected from the group consisting of hydrogen, C 1-6  alkyls, C 2-6  alkenyls, C 2-6  alkynyls, C 3-12  branched alkyls,  
  C 3-8  cycloalkyls, C 1-6  substituted alkyls, C 2-6  substituted alkenyls, C 2-6  substituted alkynyls, C 3-8  substituted cycloalkyls, aryls substituted aryls, aralkyls, C 1-6  heteroalkyls, substituted C 1-6  heteroalkyls, C 1-6  alkoxy, phenoxy and C 1-6  heteroalkoxy; and  
 R 18  is selected from the group consisting of hydrogen, C 1-6  alkyls, C 2-6  alkenyls, C 2-6  alkynyls, C 3-12  branched alkyls, C 3-8  cycloalkyls, C 1-6  substituted alkyls, C 2-6  substituted alkenyls, C 2-6  substituted alkynyls, C 3-8  substituted cycloalkyls, aryls substituted aryls, aralkyls, C 1-6  heteroalkyls, substituted C 1-6  heteroalkyls, C 1-6  alkoxy, phenoxy and C 1-6  heteroalkoxy, NO 2 , haloalkyl and halogen; and  
 
         t and q are individually selected positive integers from about 1 to about 4.  
       
     
     
         20 . The compound of  claim 1  wherein L 3  and L 4  are independently selected from the group consisting of:  
         —C(O)CR 30 R 31 OCR 32 R 33 C(O)—;  —C(O)CR 30 R 31 NR 34 CR 32 R 33 C(O)—;  —C(O)CR 30 R 31 SCR 32 R 33 C(O)—, or  —C(O)(CR 30 R 31 ) n C(O)—;  wherein: 
 R 30-34  are independently selected from H, C 1-6  alkyl, C 2-6  alkenyl, C 2-6  alkynyl, C 1-6  heteroalkyl or aryl, and  
 n is a positive integer from about 2 to about 3.  
   
     
     
         21 . A compound of  claim 1 , comprising the formula:  
       
         
           
           
               
               
           
         
         where A 1  is a leaving group.  
       
     
     
         22 . A compound of  claim 1 , selected from the group consisting of:  
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         wherein A 1  is a leaving group.  
       
     
     
         23 . The compound of  claim 1 , wherein A 1  is a leaving group selected from the group consisting of  
       
         
           
           
               
               
           
         
       
     
     
         24 . The compound of  claim 23 , wherein A is:  
       
         
           
           
               
               
           
         
       
     
     
         25 . The compound of  claim 1 , wherein A is selected from the group consisting of maleimidyl, vinyl sulfonyl, hydroxy, amino, carboxy, mercapto, hydrazide, and carbazate functional groups.  
     
     
         26 . The compound of  claim 1 , wherein said terminal branching group comprises the formula:  
       
         
           
           
               
               
           
         
         wherein: 
 Y 5  is O, S or NR 46 ;  
 L 6  is a bifunctional linker selected from the same group as that which defines L 1 ;  
 L 8  is a bifunctional linker selected from the same group as that which defines L 3 ;  
 R 40 -R 46  may be the same or different and are selected from the group consisting of hydrogen, C 1-6  alkyls, C 2-6  alkenyl, C 2-6  alkynyl, C 3-19  branched alkyls, C 3-8  cycloalkyls,  
 
         C 1-6  substituted alkyls, C 2-6  substituted alkenyls, C 2-6  substituted alkynyls, C 3-8  substituted cycloalkyls, aryls, substituted aryls, aralkyls, C 1-6  heteroalkyls, substituted C 1-6  heteroalkyls, C 1-6  alkoxy, phenoxy and C 1-6  heteroalkoxy, 
 j, j′, k and k′ are each independently 0 or a positive integer,  
 q is 0 or 1;  
 g, h, v and w are independently selected positive integers;  
 R 50  is selected from the group consisting of substantially non-antigenic polymer residues, C 1-6  alkyls, C 2-6  alkenyl, C 2-6  alkynyl, and aralkyls, and  
                     
 
         wherein: 
 L 7  is a bifunctional linker selected from the same group as that which defines L 1 ;  
 L 9  is a bifunctional linker selected from the same group as that which defines L 3 ; and  
 R 60  is selected from the group consisting of substantially non-antigenic polymer residues, C 1-6  alkyls, C 2-6  alkenyl, C 2-6  alkynyl, and aralkyls.  
 
       
     
     
         27 . A compound of  claim 26 , comprising the structure:  
       
         
           
           
               
               
           
         
       
     
     
         28 . A compound of  claim 26 , comprising the structure:  
       
         
           
           
               
               
           
         
       
     
     
         29 . A compound of  claim 1 , selected from the group consisting of  
       
         
           
           
               
               
           
         
       
     
     
         30 . A compound of  claim 1 , selected from the group consisting of  
       
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         
           
           
               
               
           
         
         wherein A is a leaving group.  
       
     
     
         31 . A method of preparing a polymer conjugate, comprising reacting a compound of the formula:  
       
         
           
           
               
               
           
         
         wherein: 
 A 1  is a leaving group;  
 R 1  and R 2  are independently selected from the group consisting of substantially non-antigenic polymer residues, H, C 1-6  alkyls, C 2-6  alkenyl, C 2-6  alkynyl, aralkyls, and terminal branching groups, provided both R 1  and R 2  are not H;  
 Z is selected from among moieties actively transported into a target cell, hydrophobic moieties, bifunctional linking moieties and combinations thereof;  
 Y 1-3  may be the same or different and are selected from among O, S or NR 11 ;  
 L 1  and L 2  may be the same or different bifunctional linkers;  
 R 3 -R 11 , R 24  and R 25  may be the same or different and are selected from the group consisting of hydrogen, C 1-6  alkyls, C 2-6  alkenyl, C 2-6  alkynyl, C 3-19  branched alkyls, C 3-8  cycloalkyls, C 1-6  substituted alkyls, C 2-6  substituted alkenyls, C 2-6  substituted alkynyls, C 3-8  substituted cycloalkyls, aryls, substituted aryls, aralkyls, C 1-6  heteroalkyls, substituted C 1-6  heteroalkyls, C 1-6  alkoxy, phenoxy and C 1-6  heteroalkoxy;  
 L 3  and L 2  may be the same or different and are selected from:  
   —C(O)(CR 30 R 31 )Y 15 (CR 32 R 33 )C(O)— or  —C(O)(CR 30 R 31 )(CR 32 R 33 )C(O)— wherein:    Y 15  is selected from O, S, NR 34  or CH 2 , and    R 30-34  may be the same or different and are selected from H, alkyl, alkenyl, alkynyl, heteroalkyl or aryl;    
 
         with an amino-containing, biologically active agent under conditions sufficient to form  
         
           
             
             
                 
                 
             
           
         
         wherein A 2  is a residue of an amine containing biologically active agent.  
       
     
     
         32 . A method of preparing a bicine-based polymer transport system, comprising: 
 a) reacting a blocked bifunctional linker with an anhydride to form an extended blocked bifunctional spacer of the formula:                          b) attaching the blocked bifunctional spacer to each hydroxyl of an acid protected bicine molecule;    c) deblocking the resultant intermediate and reacting it with an activated polymer under basic coupling conditions, and    d) deprotecting the bicine acid and thereafter activating the acid with a suitable activating group under coupling conditions.    
     
     
         33 . A method of treatment, comprising administering to a mammal in need thereof, an effective amount of a compound of  claim 1 , wherein A is a residue of a biologically active agent.  
     
     
         34 . The compound of  claim 1 , wherein A is a biologically active moiety selected from the group consisting of oligonucleotides, proteins, enzymes and organic compounds.