Mixture-Based Screening of Focused Combinatorial Libraries by NMR: Application to the Antiapoptotic Protein hMcl-1

We report on an innovativeligand discovery strategy based on proteinNMR-based screening of a combinatorial library of & SIM;125,000compounds that was arranged in 96 distinct mixtures. Using sensitivesolution protein NMR spectroscopy and chemical perturbation-basedscreening followed by an iterative synthesis, deconvolutions, andoptimization strategy, we demonstrate that the approach could be usefulin the identification of initial binding molecules for difficult drugtargets, such as those involved in protein-protein interactions.As an application, we will report novel agents targeting the Bcl-2family protein hMcl-1. The approach is of general applicability andcould be deployed as an effective screening strategy for de novo identificationof ligands, particularly when tackling targets involved in protein-proteininteractions.

Automated summary

We introduce an innovative ligand discovery strategy using protein NMR-based screening of a combinatorial library of about 125,000 compounds arranged in 96 mixtures. This approach, based on sensitive solution protein NMR spectroscopy and chemical perturbation-based screening, followed by iterative synthesis, deconvolutions, and optimization, is shown to be useful in identifying initial binding molecules for difficult drug targets, such as those involved in protein-protein interactions. As an application, we report on the discovery of novel agents targeting the Bcl-2 family protein hMcl-1. This strategy has broad applicability and can be an effective screening approach for de novo ligand identification, especially for targets involved in protein-protein interactions.