Thalidomide and its analogs are molecular glues (MGs) that lead to targeted ubiquitination and degradation of key cancer proteins via the cereblon (CRBN) E3 ligase. Here, we develop a direct-to-biology (D2B) approach for accelerated discovery of MGs. In this platform, automated, high throughput, and nano scale synthesis of hundreds of pomalidomide-based MGs was combined with rapid phenotypic screening, enabling an unprecedented fast identification of potent CRBN-acting MGs. The small molecules were further validated by degradation profiling and anti-cancer activity. This revealed E14 as a potent MG degrader targeting IKZF1/3, GSPT1 and 2 with profound effects on a panel of cancer cells. In a more generalized view, integration of automated, nanoscale synthesis with phenotypic assays has the potential to accelerate MGs discovery.

Berlin Institute of Health Berlin Germany

Department of Hematology Oncology and Cancer Immunology Charité Universitätsmedizin Berlin corporate member of Freie Universität Berlin and Humboldt Universität zu Berlin Berlin Germany

Department of Internal Medicine 3 University Hospital Ulm 89081 Ulm Germany

German Cancer Consortium Heidelberg Germany

Institute of Molecular and Translational Medicine Faculty of Medicine and Dentistry and Czech Advanced Technology and Research Institute Palackӯ University in Olomouc Olomouc Czech Republic

Max Delbrück Center for Molecular Medicine Berlin Germany

University of Groningen Department of Drug Design A Deusinglaan 1 9713 AV Groningen The Netherlands

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Precipitation-first synthesis and deep profiling of a 1200-member acrylamide library for covalent drug discovery