Three-step pretargeting methods and compounds

Abstract

Methods, compounds, compositions and kits that relate to pretargeted delivery of diagnostic and therapeutic agents are disclosed. In particular, three-step pretargeting methods are described.

What is claimed is:  
     
         1 . A method of enhancing active agent localization at a target site in a mammalian recipient, which method comprises: 
 administering to the recipient a first conjugate comprising a targeting moiety and a biotin, whereupon the first conjugate localizes to the target site;    administering to the recipient avidin or streptavidin; and    thereafter administering to the recipient a second conjugate comprising biotin, a linker resistant to biotimidase cleavage and an active agent, wherein second conjugate localization at the target site is enhanced as a result of prior localization of the first conjugate.    
     
     
         2 . A method of  claim 1  wherein the targeting moiety is proteinaceous.  
     
     
         3 . A method of  claim 1  wherein the targeting moiety is an oligonucleotide, a peptide, a polypeptide, a cytokine, a monoclonal antibody, a monovalent fragment thereof.  
     
     
         4 . A method of  claim 3  wherein the monoclonal antibody is a human, a humanized or a chimeric monoclonal antibody.  
     
     
         5 . A method of  claim 3  wherein the monoclonal antibody or fragment thereof is reactive with an antigen recognized by the antibody NR-LU-10.  
     
     
         6 . A method of  claim 1  wherein the active agent is selected from the group consisting of radionuclides, chemotherapeutic drugs, anti-tumor agents and toxins.  
     
     
         7 . A method of  claim 6  wherein the active agent is a radionuclide selected from the group consisting of Re-186, Re-188, Tc-99m, Y-90, At-211, Pb-212, Bi-212, Sm-153, Eu-169, Lu-177, Cu-67, Rh-105, In-111, Au-198, I-123 and I-131.  
     
     
         8 . A method of  claim 6  wherein the active agent is a cytokine or a lectin inflammatory response promoter.  
     
     
         9 . A method of  claim 1  wherein the step of administering the second conjugate is conducted by intralesional or intraarterial injection.  
     
     
         10 . A method of  claim 9  wherein the second conjugate is administered via an artery supplying target site tissue.  
     
     
         11 . A method of  claim 9  wherein the second conjugate is administered via an artery selected from the group consisting of hepatic artery, carotid artery, bronchial artery and renal artery.  
     
     
         12 . A method of  claim 1  wherein the second conjugate is administered intravenously.  
     
     
         13 . A method of  claim 1  wherein the second conjugate comprises a biotin-DOTA compound of the following formula:  
       
         
           
           
               
               
           
         
       
       wherein a linker L is selected from the group comprising: 
 1) a D-amino acid-containing linker of the formula  
                     
 2) a linker of the formula  
                     
 3) a linker of the formula  
                     
 4) a linker of the formula  
                     
  the group comprising: 
 a) —NH—CO—(CH 2 ) n —O—;  
 b) —NH—;  
                     
 d) —NH—CS—NH—; and  
 e) —NH—CO— (CH 2 ) n —NH—,  
 wherein R 1  is hydrogen, lower alkyl; lower alkyl substituted with one or more hydrophilic groups including (CH 2 ) m —OH, (CH 2 ) m —OSO 3 , (CH 2 ) m —SO 3 , and  
                     
 , where m is 1 or 2; glucuronide-substituted amino acids; or other glucuronide derivatives;  
 
 R 2  is hydrogen; lower alkyl; substituted lower alkyl having one or more substituents selected from the group comprising hydroxy, sulfate, and phosphonate; or a hydrophilic moiety;  
 R 3  is hydrogen; an amine; a lower alkyl; a hydroxy-, sulfate- or phosphonate-substituted lower alkyl; a glucuronide; or a glucuronide-derivatized amino acid;  
 R 4  is hydrogen, lower alkyl or  
                     
 R′ is hydrogen; —(CH 2 ) 2 —OH or a sulfate or phosphonate derivative thereof; or  
                     
  ;  
 R″ is a bond or —(CH 2 ) n —CO—NH—; and  
 n ranges from 0-5.  
 
     
     
         14 . A method of  claim 13  wherein L is a D-amino acid-incorporating linker of the formula  
       
         
           
           
               
               
           
         
       
     
     
         15 . A method of  claim 14  wherein R 1  is CH 3  and R 2  is H.  
     
     
         16 . A method of  claim 13  wherein L is a linker of the formula  
       
         
           
           
               
               
           
         
       
       .  
     
     
         17 . A method of  claim 16  wherein R 3  is hydrogen; R 4  is CH 3 ; and n is 4.  
     
     
         18 . A method of  claim 16  wherein R 3  is hydrogen; R 4  is CH 3 ; and n is 0.  
     
     
         19 . A method of  claim 16  wherein R 3  is hydrogen; R 4  is  
       
         
           
           
               
               
           
         
       
       ; and n is 4.  
     
     
         20 . A method of  claim 13  wherein L is a linker of the formula  
       
         
           
           
               
               
           
         
       
       , wherein L′ is selected from the group comprising: 
 a) —NH—CO—(CH2) n —O—;  
 b) —NH—;  
 c)  
                     
  ;  
 d) —NH—CS—NH—; and  
 e) —NH—CO—(CH 2 ) n —NH— or a bis-DOTA derivative thereof.