PO.ET09.04 · 实验与分子治疗
Rational discovery of novel molecular glues
作者与单位
摘要 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.
利益披露 Disclosure
S. Woehrle,
EMD Serono, Inc., The healthcare business of Merck KGaA, Darmstadt, Germany Employment.
A. Blum,
EMD Serono, Inc., The healthcare business of Merck KGaA, Darmstadt, Germany Employment.
B. Blume,
EMD Serono, Inc., The healthcare business of Merck KGaA, Darmstadt, Germany Employment.
J. Bomke,
EMD Serono, Inc., The healthcare business of Merck KGaA, Darmstadt, Germany Employment.
H. Buchstaller,
EMD Serono, Inc., The healthcare business of Merck KGaA, Darmstadt, Germany Employment.
V. Dragone,
EMD Serono, Inc., The healthcare business of Merck KGaA, Darmstadt, Germany Employment.
I. Ferrara,
EMD Serono, Inc., The healthcare business of Merck KGaA, Darmstadt, Germany Employment.
A. Gambardella,
EMD Serono, Inc., The healthcare business of Merck KGaA, Darmstadt, Germany Employment.
J. Gunera,
EMD Serono, Inc., The healthcare business of Merck KGaA, Darmstadt, Germany Employment.
U. Grädler,
EMD Serono, Inc., The healthcare business of Merck KGaA, Darmstadt, Germany Employment.
I. Kober,
EMD Serono, Inc., The healthcare business of Merck KGaA, Darmstadt, Germany Employment.
J. Krieger,
EMD Serono, Inc., The healthcare business of Merck KGaA, Darmstadt, Germany Employment.
B. Leuthner,
EMD Serono, Inc., The healthcare business of Merck KGaA, Darmstadt, Germany Employment.
A. Unzue Lopez,
EMD Serono, Inc., The healthcare business of Merck KGaA, Darmstadt, Germany Employment.
O. Schadt,
EMD Serono, Inc., The healthcare business of Merck KGaA, Darmstadt, Germany Employment.
C. Schindler,
EMD Serono, Inc., The healthcare business of Merck KGaA, Darmstadt, Germany Employment.
F. J. Sorrell,
EMD Serono, Inc., The healthcare business of Merck KGaA, Darmstadt, Germany Employment.
A. M. Esteves,
Instituto de Biologia Experimental e Tecnológica (iBET) Employment.
S. P. Santos,
Instituto de Biologia Experimental e Tecnológica (iBET) Employment.
R. L. Sousa,
Instituto de Biologia Experimental e Tecnológica (iBET) Employment.
M. M. S. Silva,
Instituto de Biologia Experimental e Tecnológica (iBET) Employment.
A. R. Lemos,
Instituto de Biologia Experimental e Tecnológica (iBET) Employment.
P. M. F. Sousa,
Instituto de Biologia Experimental e Tecnológica (iBET) Employment.
T. M. Bandeiras,
Instituto de Biologia Experimental e Tecnológica (iBET) Employment.
M. Cuomo,
EMD Serono, Inc., The healthcare business of Merck KGaA, Darmstadt, Germany Employment.