Extended tethering approach for rapid identification of ligands

Abstract

The invention concerns a method for rapid identification and characterization of binding partners for a target molecule, and for providing binding partners with improved binding affinity. More specifically, the invention concerns an improved tethering method for the rapid identification of at least two binding partners that bind near one another to a target molecule.

1 - 74 . (canceled)  
     
     
         75 . A method of synthesizing a non-peptide small molecule comprising reacting a first ligand with a second ligand, wherein 
 (a) the first ligand has inherent binding affinity for a first site of interest on a Target Biological Molecule (TBM) having a reactive nucleophile; and    (b) the second ligand has been identified from among a library of ligand candidates to have inherent binding affinity for a second site of interest on said TBM by screening said library with a complex formed between a reactive derivative of said first ligand and the nucleophile of said TBM resulting in the formation of an irreversible covalent bond between said complex and said second ligand:    
     
     
         76 . The method of  claim 75  wherein said first ligand has been identified from among a library of ligand candidates by screening said library with said TBM under conditions resulting in the formation of a reversible covalent bond between said nuclephile and said first ligand  
     
     
         77 . The method of  claim 75  wherein said nucleophile is selected from the group consisting of thiol, protected thiol, reversible disulfide, hydroxyl, protected hydroxyl, amino, protected amino, carboxyl and protected carboxyl groups.  
     
     
         78 . The method of  claim 75  wherein said first and second ligands are covalently linked to one another through a disulfide bond.  
     
     
         79 . The method of  claim 75  wherein said first and second ligands are directly fused to one another.  
     
     
         80 . The method of  claim 75  wherein said small molecule consists essentially of said first and second ligands, covalently linked through a disulfide bond.  
     
     
         81 . The method of  claim 78  further comprising the step of synthesizing derivatives of said small molecule.  
     
     
         82 . The method of  claim 81  wherein the disulfide bond covalently linking said first and second ligands to one another is replaced by a different covalent linkage.  
     
     
         83 . The method of  claim 82  wherein the different linkage is a straight chain, branched chain or aromatic linker.  
     
     
         84 . The method of  claim 83  wherein the different linkage is a straight chain linker of at least two atoms in length.  
     
     
         85 . The method of  claim 83  wherein the different linkage is a straight chain linker of at least four atoms in length.  
     
     
         86 . The method of  claim 83  wherein the different linkage is a straight chain linker of at least 12 atoms in length.