A high throughput compatible workflow for the biochemical identification and characterisation of molecular glues.

Abstract

Molecular glues are an emerging modality which induces or enhances an interaction between two proteins. Molecular glues can target proteins via proximity-induced degradation or sequestration and can, therefore, provide opportunities for therapeutic intervention to targets that cannot be modulated by traditional small molecule approaches. Due to their modest molecular weight, molecular glues may not encounter the bioavailability issues associated with PROTACs. Characterisation of molecular glues in hit finding and hit optimisation settings can be challenging, as both the affinity of the glue for the target protein and the resulting improvement in affinity between the proteins of interest need to be assessed in parallel. Here, we propose and validate a workflow to derive both key parameters from a classic concentration response experiment. Furthermore, we provide a method for the rational determination of optimum biochemical assay conditions to identify and characterise molecular glues.