Branched discrete peg constructs

Abstract

Disclosed are general and “substantially pure” branched discrete polyethylene glycol constructs useful in attaching to a variety of biologically active groups, for example, preferential locators, as well as biologics like enzymes, for use in diagnostics, e.g. imaging, therapeutics, theranostics, and moieties specific for other applications. In its simplest intermediate state, a branched branched discrete polyethylene glycol construct is terminated at one end by a chemically reactive moiety, “A”, a group that is reactive with a biologic material that creates “A”, which is a biologically reactive group, connected through to a branched core (BC) which has attached at least two dPEG-containing chains, indicated by the solid line, , having terminal groups, which can be charged, non-reactive or reactable moieties and containing between about 2 and 64 dPEG residues.

1 . (canceled) 
     
     
         2 . A substantially pure compound, represented by: 
       
         
           
           
               
               
           
         
         wherein: 
         (a) A is a biologically active group or a chemically reactive or reactable moiety; 
         (b) the wavy line,   is a linear discrete polyethylene glycol chain containing between about 4 and 48 discrete ethylene oxide residues optionally substituted with N, S, Si, Se, or P, aryl groups, or alkyl groups; and having chemically reactive or reactable end groups that are independently reactable; and optionally having branching side chains which can contain diagnostic or therapeutic groups and being cleavable; 
         (c) each BC group may have, independently,   or   attached; and 
         (d)   and   are part of 
       
       
         
           
           
               
               
           
         
       
       respectively, where the solid lines  , are discrete polyethylene glycol containing chains that have between about 8 and 72 ethylene oxide residues and have a terminal moiety, wherein the terminal moiety can be neutral or charged. 
     
     
         3 . A substantially pure compound, represented by: 
       
         
           
           
               
               
           
         
         wherein: 
         (a) A is a biologically active group or a chemically reactive or reactable moiety; 
         (b) the wavy line,  , is a linear discrete polyethylene glycol chain containing between about 4 and 48 discrete ethylene oxide residues optionally substituted with N, S, Si, Se, or P, aryl groups, or alkyl groups; and having chemically reactive or reactable end groups that are independently reactable; and optionally having branching side chains which can contain diagnostic or therapeutic groups and being cleavable; 
         (c) BC is a trifunctional core and may be the same as BC 1 , or BC 2 ; where BC 1  and BC 2  are not the same; and BC 1  and BC 2  independently may have attached either   or  , where each is different for attachment to BC 1  and BC 2 , respectively, but can be either   or  ; and 
         (d)   and   are part of 
       
       
         
           
           
               
               
           
         
       
       respectively, where the solid lines,  , are discrete polyethylene glycol containing chains that have between about 8 and 72 ethylene oxide residues and have a terminal moiety, wherein the terminal moiety can be neutral or charged. 
     
     
         4 . A substantially pure compound represented by: 
       
         
           
           
               
               
           
         
         wherein: 
         a) A is one or more of 
       
       
         
           
           
               
               
           
         
         b) the wavy line,  , is a linear discrete polyethylene glycol chain containing between about 4 and 48 discrete ethylene oxide residues optionally substituted with N, S, Si, Se, or P, aryl groups, or alkyl groups; and having chemically reactive or reactable end groups that are independently reactable; and optionally having branching side chains which can contain diagnostic or therapeutic groups and being cleavable; 
         c) BC is a branching core containing a carbon core with 3 or 4 reactable sites, one of said carbon reactable sites being unique, for attachment of   and the other 2 or 3 carbon reactable sites being independently reactive or reactable moieties, for attachment of  , respectively; and 
         d)   and   are part of 
       
       
         
           
           
               
               
           
         
       
       respectively, where the solid lines,  , are discrete polyethylene glycol containing chains that have between about 8 and 72 ethylene oxide residues and have a terminal moiety, wherein the terminal moiety can be neutral or charged. 
     
     
         5 . A substantially pure compound, represented by: 
       
         
           
           
               
               
           
         
         wherein: 
         a) each A 1 , A 2 , and A 3  is a biologically active group or a chemically reactive or reactable moiety and, independently may be the same or different; 
         b) the wavy line,  , is a linear discrete polyethylene glycol chain containing between about 4 and 48 discrete ethylene oxide residues optionally substituted with N, S, Si, Se, or P, aryl groups, or alkyl groups; and having chemically reactive or reactable end groups that are independently reactable; and optionally having branching side chains which can contain diagnostic or therapeutic groups and being cleavable; 
         c) any BC, independently, may be the same or different and each BC is a branching core containing a carbon core with 3 or 4 reactable sites, one of said carbon reactable sites being unique, for attachment of   and the other 2 or 3 carbon reactable sites being independently reactive or reactable moieties, for attachment of  , respectively, which is  . 
       
     
     
         6 . A method of externally imaging an antibody, antibody fragment, or a peptide attached to an externally imageable radiolabel, which comprises attaching said imageable radiolabel to and said antibody, antibody fragment, or a peptide to the structure of  claim 2 , wherein RG is said radiolabel and A comprises said antibody, antibody fragment, or a peptide. 
     
     
         7 . The method of  claim 6 , wherein said antibody has been generated against a tumor associated glycoprotein. 
     
     
         8 . The method of  claim 7 , wherein said antibody comprises B72.3, CC49, V59, or 3E8. 
     
     
         9 . The method of  claim 8 , wherein said antibody comprises CC49; domain deleted CC49; CC49 fragments, including Fab′; CC49 diabodies; CC49 scFv; and single chains derived therefrom. 
     
     
         10 . The method of  claim 6 , wherein said antibody comprises 3E8, domain deleted 3E8, 3E8 fragments, 3E8 diabodies, and single chains derived therefrom. 
     
     
         11 . The substantially pure compound of  claim 2 , having the formula:
 (a) H 2 N-dPEG 12 -Tris(-dPEG 12 -Tris(-dPEG 11 -m) 3 ) 3 ;   (b) MAL-dPEG 12 -Tris(-dPEG 12 -Tris(m-dPEG 11 ) 3 ) 3 ;   (c) H 2 N-dPEG 24 -Tris(-dPEG 12 -Tris(-dPEG 24 -m) 3 ) 3 ;   (d) MAL-dPEG 24 -Tris(-dPEG 12 -Tris(m-dPEG 24 ) 3 ) 3 ; or   (e) MAL-dPEG 24 -Tris(-dPEG 24 -Tris(m-dPEG 24 ) 3 ) 3 .   
     
     
         12 . A method of externally imaging an antibody, antibody fragment, or a peptide attached to an externally imagable radiolabel (RG), which comprises attaching said imagable radiolabel and said antibody, antibody fragment, or a peptide to the substantially pure compound of  claim 2 , wherein A comprises said antibody, antibody fragment, or a peptide. 
     
     
         13 . A substantially pure compound, represented by: 
       
         
           
           
               
               
           
         
         wherein: 
         a) AC is an attachment core and G is a chemically reactable moiety or a moiety useful for one or more diagnostic or therapeutic applications, and where x=1 to 10; and A is a biologically active group or a chemically reactive or reactable moiety; optionally G is a branched chain; 
         b) the wavy line,  , is a linear discrete polyethylene glycol chain containing between about 4 and 48 discrete ethylene oxide residues optionally substituted with N, S, Si, Se, or P, aryl groups, or alkyl groups; and having chemically reactive or reactable end groups that are independently reactable; and optionally having branching side chains and being cleavable; 
         c) BC is a branching core containing a carbon core with 3 or 4 reactable sites, one of said carbon reactable sites being unique and the other 2 or 3 carbon reactable sites being independently reactive or reactable moieties; and 
         d)   and   are part of 
       
       
         
           
           
               
               
           
         
       
       respectively, where the solid lines,  , are discrete polyethylene glycol containing chains that have between about 2 and 64 ethylene oxide residues and have a terminal moiety, wherein the terminal moiety can be neutral, charged or independently reactable. 
     
     
         14 . A substantially pure compound, represented by: 
       
         
           
           
               
               
           
         
         wherein: 
         a) A is a biologically active group or chemically reactive or reactable moiety; 
         b) the wavy line,  , is a linear discrete polyethylene glycol chain containing between about 4 and 48 discrete ethylene oxide residues optionally substituted with N, S, Si, Se, or P, aryl groups, or alkyl groups; and having chemically reactive or reactable end groups that are independently reactable; and optionally having branching side chains and being cleavable; 
         c) when x=1, the compound is part of a second  ; 
         d) when x=2 or 3, the aryl core is part of a unique branching core (BC), where O  A is a solid line,  , with a terminal group; and 
         e) R is any group known in the art that can be added as an additional attachment point for another A; 
         wherein, the solid line,  , is a discrete polyethylene glycol-containing chain that have between about 2 and 64 ethylene oxide residues and have a terminal moiety, wherein the terminal moiety can be neutral, charged, or independently reactable; and BC is a branching core containing a carbon core with 3 or 4 reactable sites, one of said carbon reactable sites being unique, for attachment of  , and the other 2 or 3 carbon reactable sites being independently reactive or reactable moieties, for attachment of  , respectively.