High affinity TGFbeta nucleic acid ligands and inhibitors

Abstract

Methods are described for the identification and preparation of high-affinity nucleic acid ligands to TGFβ2. Included in the invention are specific RNA ligands to TGFβ2 identified by the SELEX method. Also included are RNA ligands that inhibit the interaction of TGFβ2 with its receptor.

What is claimed is:  
     
         1 . A method of inhibiting a transforming growth factor β2 (TGFβ2) comprising contacting said TGFβ2 with a nucleic acid ligand of TGFβ2.  
     
     
         2 . The method of  claim 1 , wherein the nucleic acid ligand of TGFβ2 is a ligand comprising a ligand having a nucleotide sequence selected from the group consisting of SEQ ID NOS:21-87, 89, 91-93, 109, 111, 114-116, 118-121, 129, 131, 138, 140, 144, 146-181, 184-189, 192, and 193.  
     
     
         3 . The method of  claim 1  wherein said nucleic acid ligand is conjugated to polyethylene glycol (PEG).  
     
     
         4 . The method of  claim 3  wherein said PEG has a molecular weight of about between 10-80 K.  
     
     
         5 . The method of  claim 3  wherein said PEG has a molecular weight of about 20-45 K.  
     
     
         6 . The method of  claim 1  wherein said ligand is  
       
         
           
           
               
               
           
         
       
       wherein 
 X=PEG, and  
 LIGAND=rGrGrArGrGfUfUrAfUfUrAfCrArGrArGfUfCfUrGfUfUrArGfCfUrGfUrAfCfUfCfC-3′-3′-dT (SEQ ID NO:115), wherein rG is 2′OH G, rA is 2′OH A, fU is 2′F U and fC is 2′F C.  
 
     
     
         7 . A method for targeting a nucleic acid ligand to a site in a patient comprising TGFβ82 comprising: 
 covalently linking said nucleic acid ligand to a Non-Immunogenic, High Molecular 20 Weight Compound or Lipophilic Compound to form a Complex, and administering said Complex to said patient, whereby said nucleic acid ligand is targeted to a site in a patient comprising TGFβ2.  
 
     
     
         8 . The method of  claim 7 , wherein the nucleic acid ligand of TGFβ2 is a ligand comprising a ligand having a nucleotide sequence selected from the group consisting of SEQ ID NOS:21-87, 89, 91-93, 109, 111, 114-116, 118-121, 129, 131, 138, 140, 144, 146-181, 184-189, 192, and 193.  
     
     
         9 . The method of  claim 7  wherein said nucleic acid ligand is conjugated to polyethylene glycol (PEG).  
     
     
         10 . The method of  claim 9  wherein said PEG has a molecular weight of about between 10-80 K.  
     
     
         11 . The method of  claim 9  wherein said PEG has a molecular weight of about 20-45 K.  
     
     
         12 . The method of  claim 7  wherein said ligand is  
       
         
           
           
               
               
           
         
       
       wherein 
 X=PEG, and  
 LIGAND=rGrGrArGrGfUfUrAfUfUrAfCrArGrArGfUfCfUrGfUfUrArGfCfUrGfUrAfCfUfCfC-3′-3′-dT (SEQ ID NO:1 15), wherein rG is 2′OH G, rA is 2′OH A, fU is 2′F U and fC is 2′F C.  
 
     
     
         13 . A method for treating a TGF□2-mediated pathological conditions comprising administering a nucleic acid ligand capable of binding to TGF□2 to a patient in need thereof.  
     
     
         14 . The method of  claim 13 , wherein the nucleic acid ligand of TGFβ2 is a ligand comprising a ligand having a nucleotide sequence selected from the group consisting of SEQ ID NOS:21-87, 89, 91-93, 109, 111, 114-116, 118-121, 129, 131, 138, 140, 144, 146-181, 184-189, 192, and 193.  
     
     
         15 . The method of  claim 13  wherein said nucleic acid ligand is conjugated to polyethylene glycol (PEG).  
     
     
         16 . The method of  claim 15  wherein said PEG has a molecular weight of about between 10-80 K.  
     
     
         17 . The method of  claim 15  wherein said PEG has a molecular weight of about 20-45 K.  
     
     
         18 . The method of  claim 13  wherein said ligand is  
       
         
           
           
               
               
           
         
       
       wherein 
 X=PEG, and  
 LIGAND=rGrGrArGrGfUfUrAfUfUrAfCrArGrArGfUfCfUrGfUfUrArGfCfUrGfUrAfCfUfCfC-3′-3′-dT (SEQ ID NO: 115), wherein rG is 2′OH G, rA is 2′OH A, fU is 2′F U and fC is 2′F C.