Abstract Purpose: We developed and applied a streamlined workflow to discover and optimize cereblon (CRBN)-mediated molecular glues. Methods: We combine computational and structural approaches to define the CRBN target space with both CRBN-focused small molecule and DNA-encoded (DEL) libraries for high-throughput screening of novel molecular glues. Proximity based protein-protein interaction assays are applied to screen for induced CRBN proximity of novel glue degrader targets. Cell-based degradation methods can then drive lead optimization, with advanced biophysical and structural methodologies for structural and functional aided design. Results and Conclusions: We have identified multiple leads from rational screening approaches inducing concentration-dependent target-CRBN proximity in TR-FRET and NanoBRET assays. Hits were orthogonally confirmed to induce ternary complex formation by biophysical methods including surface plasmon resonance (SPR) and size exclusion chromatography (SEC). Cellular readouts demonstrated degradation activity using HiBiT and immunofluorescence-based methods dependent on CRBN E3 ligase expression or activity and proteasomal degradation. This integrated workflow together with proteomics technologies for on-target and off-target selectivity, is a rapid, mechanism-centric discovery platform for CRBN molecular glues across diverse target classes taking advantage of advanced but readily available methodologies. Citation Format: Simon Woehrle, Andreas Blum, Beatrix Blume, Jörg Bomke, Hans-Peter Buchstaller, Vincenza Dragone, Isabella Ferrara, Alessia Gambardella, Jakub Gunera, Ulrich Grädler, Ingo Kober, Johannes Krieger, Birgitta Leuthner, Andrea Unzue Lopez, Oliver Schadt, Christina Schindler, Fiona J. Sorrell, Ana M. Esteves, Sandra P. Santos, Raquel L. Sousa, Margarida M. Silva, Ana R. Lemos, Pedro M. Sousa, Tiago M. Bandeiras, Maria Emanuela Cuomo. Rational discovery of novel molecular glues abstract. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 5788.
Woehrle et al. (2026) studied this question.