Patent ID: 7252931

Claim:
A method for generating a combinatorial library of bi-ligand compounds that can simultaneously bind to a CL site and an SL site of an enzyme, wherein the enzyme binds a common ligand (CL) or a common ligand mimic (CL mimic) at a common ligand site (CL site) and binds a specificity ligand (SL) at an adjacent specificity ligand site (SL site); wherein an interface region is defined as the atoms of the enzyme between the CL site and SL site, and atoms of an SL, if bound to the enzyme; wherein the enzyme catalyzes a reaction involving the SL and a reactive atom of the CL; and wherein a CL reactive region is defined as the reactive atom of the CL and immediately adjacent atoms of the CL; comprising the steps of (a) identifying an atom of the interface region, comprising the steps of (1) binding a CL to the CL site of said enzyme; (2) irradiating a nucleus of an atom of the CL reactive region; and (3) identifying in a multidimensional NMR experiment an NMR cross-peak corresponding to a nucleus that is perturbed by the irradiation of the nucleus of the CL reactive region, thereby identifying an atom of the interface region; then (b) identifying an atom in the CL mimic that is proximal to the interface region, comprising the steps of (1) binding a CL mimic to the CL site of said enzyme; (2) irradiating the nucleus of the interface atom identified in step (a); and (3) identifying in a multidimensional NMR experiment an NMR cross-peak corresponding to a nucleus of the CL mimic that is perturbed by the irradiation of the interface nucleus, thereby identifying an atom of the CL mimic that is proximal to the interface region; and (c) synthesizing at least two compounds by modifying at least one atom of the CL mimic identified in step (b)(3) by attaching a substituent group to said atom, thereby generating a combinatorial library of bi-ligand compounds that can simultaneously bind to a CL site and an SL site of an enzyme.