PROTAC
(PROteolysis TArgeting Chimeras)
PROTACs provide significant therapeutic potential with possible prolonged pharmacodynamics, improved potency, and ability to target proteins previously thought of as “undruggable”. PROTACs are heterobifunctional small molecules consisting of a target binding handle bridged via a chemical linker to an E3 ligase handle which hijacks the endogenous ubiquitin machinery to target proteins, resulting in subsequent ubiquitination and degradation of your target of interest.
At o2h discovery, we have strong expertise in the synthesis of PROTACs and related chemistry, providing comprehensive solutions for PROTAC building blocks, linkers, compound libraries, and related intermediates to support Targeted Protein Degradation and integrated drug discovery programs.
protein degradation – biology
o2h biologists can support you in addressing the following questions:
Q. Is my target degraded?
Q. Is my target ubiquitinated?
Q. Do they form a ternary complex?
Q. Are they soluble and cell permeable?
Q. What is the binary affinity for POI and E3 ligase?
Q. What is the phenotypic consequence of target degradation?
Our biology evaluation suite is designed to provide comprehensive insights into ternary complex formation containing the degrader molecule, with tagged E3 ligase and the target of interest, transiently transfected into suitable cell lines. This is a crucial step in targeted protein degradation and helps in optimising the degrader molecule. We also study the degradation kinetics of endogenous targets using CRISPR gene editing following PROTAC treatment. Cellular dose response curves can be monitored in real time allowing accurate determination of DC50 values. o2h utilises SPR studies to understand binary affinities and ternary complex formation of degrader molecules in a biophysical setting. In addition, we can measure phenotypic consequences of target degradation, dissect alterations in downstream signalling and conduct functional studies.
For a complete understanding of your compounds, our ADME profiling covers essential parameters like ChamelogK, cellular permeability and solubility, ensuring they meet the necessary criteria.
Case Study
(PROTAC-induced BRD4-VHL ternary complex formation and BRD4 protein degradation. HEK292 cells transfected with NanoLuc-BRD4 donor plasmid and Halo-Tag-VHL acceptor plasmid at 1:100 ratios. Subsequently cells treated with 1uM MZ1 or AT1 PROTAC and compared with untreated DMSO control.
(A, B) Indicates ternary complex formation at 6h (C, D) BRD4-BD2 protein levels at 6h (E, F) real-time ternary complex formation with BRD4-B2/VHL following PROTAC treatment (G, H) real-time kinetics of BRD4-BD2 protein degradation following PROTAC treatment. MG132, a potent, reversible and cell permeable proteasomal inhibitor rescues BRD4 degradation in all condition.)
If you want to Jump-start your PROTAC based drug discovery program (targeted protein degradation), or would like to get a quote, write to us at discovery@o2h.com
