A Complete Mechanistic Framework for Molecular Glue Characterization

Abstract

The use of molecular glues to modulate protein–protein interactions has emerged as a promising approach to drug previously intractable targets. Despite strong interest in this modality, a comprehensive theoretical framework is lacking. We have established a complete mechanistic analysis which enables the determination of molecular glues’ binding and cooperativity values from a simple biochemical matrix experiment. We validated the model using numerical integration and experimental data from fluorescence resonance energy transfer and fluorescence polarization assays. As the derived models enable the determination of all thermodynamic parameters of any molecular glue, they have significant applications in drug discovery, particularly for molecular glue optimization and characterization of neo-substrates.