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PROTACs & Molecular Glues

Advanced proteolysis-targeting solutions for next-generation drug discovery and development

PROTACs (PROteolysis-TArgeting Chimeras) artificially hijack the components of the UPS to degrade a target protein. PROTAC drugs are hetero-bifunctional small molecules that contain two functional ligands connected via a linker; one ligand binds to a target protein and the other ligand binds to an E3 ligase.

Bringing these two entities into proximity theoretically leads to polyubiquitylation and proteasomal degradation of the target protein. However, given the complexity this scenario does not always play out, and the PROTAC discovery strategy faces several challenges and pitfalls.

To address these challenges, LifeSensors developed high-throughput tools for monitoring both polyubiquitylation and degradation of a target protein. Measuring ubiquitylation of a protein in a plate-based format accelerates PROTAC-based drug discovery.

We have developed several platforms that enable rapid, quantitative monitoring of in vitro as well as cellular ubiquitylation.

High-Throughput

Rapid screening and analysis capabilities

Quantitative

Precise measurement and data analysis

Versatile

In vitro and cellular applications

PROTAC® Degradation Assays

Comprehensive degradation monitoring and analysis for your target proteins using our validated assay platforms.

PROTAC® Ubiquitination Assays

High-throughput ubiquitination monitoring to accelerate your PROTAC development pipeline.

Target Exploration and Validation

Identify and validate optimal protein targets for your therapeutic development programs.

Novel E3 Ligases for PROTAC®

Access our proprietary library of E3 ligases to expand your PROTAC design possibilities.

PROTAC Ternary Complex Assays

Characterize and optimize ternary complex formation for improved PROTAC efficacy.

PROTAC for Cancer

Specialized PROTAC solutions targeting oncogenic proteins for cancer therapeutics.

PROTAC for Neuroscience

Targeted protein degradation approaches for neurological disease applications.

Explore Our Research Publications

Discover the latest findings and innovations in PROTAC technology

PROTACs & Molecular Glues

PROTACs (Proteolysis-targeting chimeric molecules) artificially hijack the components of the UPS to degrade a target protein. PROTAC drugs are hetero-bifunctional small molecules that contain two functional ligands connected via a linker; one ligand binds to a target protein and the other ligand binds to an E3 ligase. Bringing these two entities into proximity theoretically leads to polyubiquitylation and proteasomal degradation of the target protein. However, given the complexity this scenario does not always play out, and the PROTAC discovery strategy faces several challenges and pitfalls.

To address these challenges, LifeSensors developed high-throughput tools for monitoring both polyubiquitylation and degradation of a target protein. Measuring ubiquitylation of a protein in a plate-based format accelerates PROTAC-based drug discovery. We have developed several platforms that enable rapid, quantitative monitoring of in vitro as well as cellular ubiquitylation.

PROTAC® Degradation Assays

PROTACs (PROteolysis TArgeting Chimeras) are heterobifunctional molecules with a distinct ligand that targets a specific E3 ligase which is tethered by another ligand specific for the target protein, using an optimized chemical linker. This allows to exploit cell’s own degradation machinery (Ubiquitin proteasome system) to degrade target proteins. PROTAC approach has shown great promise to degrade proteins that are previously deemed extremely difficult to target but their expansion to vast undruggable proteome remains elusive due to lack of efficient workflow to understand structure-activity relationship and rationally design potent PROTACs.

LifeSensors offer a comprehensive evaluation of PROTAC mediated ubiquitination and degradation using traditional western blotting, reporter gene assays (fluorescent proteins, HiBiT) and LifeSensors’ high-throughput ubiquitination and degradation assays. Our targeted protein degradation team has a strong track record in developing preclinical PROTAC programs starting with identification of ligands for target, ligands for E3 ligases, designing model PROTACs, establishing ubiquitination, degradation, and lead optimization. During the optimization stage we offer collaboration with chemistry teams to rationally design a PROTAC that has superior ubiquitination and degradation profiles that can translate into reliable clinical candidates.

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PROTAC® Ubiquitination Assays

PROTACs (PROteolysis TArgeting Chimeras) are heterobifunctional molecules with a distinct ligand that targets a specific E3 ligase which is tethered by another ligand specific for the target protein, using an optimized chemical linker. Current PROTAC discovery heavily relies on monitoring the Ternary complex formation using biophysical and biochemical approaches like TR-FRET proximity ligand assays and SPR that does not monitor true function of PROTAC i.e., PROTAC mediated ubiquitination. Designing PROTACs that harness the protein’s intrinsic properties, such as exposed lysines that in turn result in efficient ubiquitination, is crucial to develop potent degraders. Most current PROTACs utilize only a few E3 ligases, which ultimately hinders the vast expansion of the undruggable proteasome. LifeSensors offers a comprehensive evaluation of PROTAC mediated ubiquitination and degradation using customized assays. The in vitro ubiquitination platform was designed to monitor multiple variants of PROTAC with variable linkerology, exit vectors, variable ligands for an E3 ligase, or targets to evaluate the best combination that results in robust ubiquitination. We offer collaboration with chemistry teams to rationally design PROTACs that have superior ubiquitination efficiency that can translate into superior degradation in vivo.

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Target Exploration and Validation

PROTACs (PROteolysis TArgeting Chimeras) are heterobifunctional molecules with a distinct ligand that targets a specific E3 ligase which is tethered by another ligand specific for the target protein, using an optimized chemical linker. This allows for exploitation of the cell’s own degradation machinery (Ubiquitin proteasome system) to degrade target proteins. Validation of a novel target’s ability to degrade with subsequent observation of the predicted phenotype is crucial for PROTAC drug discovery.

At LifeSensors, we offer a comprehensive overview of endogenous target protein expression, generate cell lines with a degrader tag (dTAG, BromoTAG®) on target protein of interest and induce selective degradation with TAGged protein of interest that can elucidate the role of the target in physiological function or a disease mechanism to produce a relevant phenotype. Successful validation of a target followed by the design of chemical modulators like PROTACs that can selectively modulate the endogenous target is largely considered direct evidence of TPD campaign.

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Novel E3 Ligases for PROTAC ®

Proteolytic targeting chimeras (PROTACs) are heterobifunctional small molecules consisting of an E3 ligase recruiting molecule and a target protein ligand, tethered by chemical linkers. There are more than 600 E3 ligases in the human proteome, but only a few ligases are currently being used for targeted protein degradation, such as Cereblon (CRBN), Von Hippel Lindau (VHL), MDM2, and IAPs. While CRBN and VHL have shown broad applicability for PROTAC applications, the lack of tractable ligands for novel E3 ligases and the lack of reliable screening methods limit the expansion of PROTACs to new E3 ligases. Despite the great success of traditional drug targets (such as kinases) and “undruggable” targets (such as RAS family proteins), there are still many targets that remain elusive and do not benefit from PROTACs.

Lifesensors has developed a suite of assays that can screen for novel ligands and design PROTACs recruiting novel ligases and aid in the discovery of a new generation of PROTACs. Currently, LifeSensors has a library of ~50 ligases that can be explored for PROTAC applications based on target protein compartmentalization and tissue specificity for efficient degradation. Our Novel E3 ligases for PROTAC service include high-throughput screening for identifying novel ligands, characterizing ligand affinity to novel E3 ligase, demonstrating selectivity, PROTAC design, and evaluating target degradation.

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Advantages of Choosing LifeSensors for your PROTAC Drug Discovery

PROTAC Ternary Complex Assays

PROTACs (PROteolysis Targeting Chimeras) are heterobifunctional molecules that combine a target-binding warhead with an E3 ligase-binding moiety, resulting in the formation of a ternary complex between the target protein and the E3 ligase to induce ubiquitination and degradation. It is increasingly clear that the length and composition of the linker connecting the two ligands plays a crucial role in the physicochemical properties and biological activity of PROTACs. The PROTAC field is rapidly evolving and is currently undergoing an important transition from synthetically manageable alkyl and polyethylene glycol linkers to more complex functional linkers. Traditionally, ternary complex formation has been assessed using cell-free proximity ligand assays such as TR-FRET, ALPHA screening assays, and SPR-based methods.

Studies in recent years have shown that protein degradability is strongly influenced by intrinsic characteristics of the protein, especially the protein’s endogenous “Ubiquitination Potential”. Hence correlating Ternary complex formation with ubiquitination potential is crucial in accurately characterizing PROTAC efficiency and helping chemists rationally design. LifeSensors offers wide array of services to validate PROTAC ternary complex, PROTAC ubiquitination and determining binding affinities (SPR).

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PROTAC for Cancer

Remarkable advances have been made in cancer targeted therapies with small molecule inhibitors and monoclonal antibodies. Drug resistance and off-target effects have limited their use making researchers seek more effective targeted therapies. With the development of proteolytic targeting chimeras (PROTACs), the utilization of protein degradation mechanisms as a therapeutic strategy to degrade oncogenic proteins is undergoing revolutionary advances. The first oral PROTACs (ARV-110 and ARV-471) showed encouraging results in clinical trials with prostate and breast cancer patients sparking greater enthusiasm for developing next generation of PROTACs. PROTACs are heterobifunctional structures consisting of ligands that bind the protein to be degraded and ligands to E3 ubiquitin ligases. PROTAC-mediated bridging between the protein of interest and E3 ligase promotes protein ubiquitination and its proteasomal degradation. There is a greater emphasis need on optimization of the ligases used in the design of PROTACs for specific tumor tissues to develop targeted therapeutics.

LifeSensors suite of assays are designed to study PROTAC-mediated ubiquitination and degradation of oncogenes of interest, identify suitable ligases and provide mechanistic insights that help chemists rationally design potent PROTACs. LifeSensors offers comprehensive solutions for all your needs related to PROTAC drug discovery.

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PROTAC for Neuroscience

A wide range of neurodegenerative diseases affect the central nervous system (CNS) and lead to impaired sensory, motor, and cognitive processes. This impairment can lead to permanent damage to neuronal health and eventual neuronal death. The complex disease pathophysiology and poorly studied underlying mechanisms have led to an increase in therapeutic efforts, which have historically been proven to be difficult. Proteolytic Targeted Chimera (PROTAC®) protein degraders open up new and exciting opportunities for degrading disease-causing mutants and protein aggregates, starting with tauopathies and synucleinopathies. Some hallmarks of the new generation of PROTACs are improved pharmacology to cross the blood-brain barrier (BBB), increased catalytic activity to break down aggregates to prevent diffusion to healthy neurons, and the ability to adapt to new delivery mechanisms. There is ample evidence that large protein aggregates are proteasome-resistant, so PROTACs that selectively drive degradation through the autophagosomal mechanism can significantly improve therapeutic outcomes.

It is of great significance to study novel E3 ligases compared to traditional E3 ligases to generate polyubiquitin chains that can promote both proteasomal and lysosomal degradation. Lifesensors has developed custom high-throughput assays to study PROTAC-mediated ubiquitination and degradation of protein aggregates to provide mechanistic insights that help chemists design reliable SARs.

Tauopathies are neurodegenerative disorders marked by the buildup of abnormal tau proteins, ultimately resulting in neuronal demise. A recent innovative study sheds light on the pathogenesis of tauopathies and offers potential therapeutic strategies targeting tau protein degradation.¹ Explore groundbreaking research on tauopathies conducted by Silva, M. C., et al., as presented in their paper “Targeted degradation of aberrant tau in frontotemporal dementia patient-derived neuronal cell models.”

Another groundbreaking study by Wang W, et al. presenting a PROTAC degrader aimed at selectively degrading tau proteins implicated in Alzheimer’s disease (AD) and related tauopathies. Their small-molecule PROTAC, C004019, effectively promotes tau clearance both in vitro and in vivo. This study demonstrates significant reductions in tau levels in the brains of wild-type, hTau-transgenic, and 3xTg-AD mice following administration of C004019. This reduction in tau correlates with improvements in synaptic and cognitive functions, highlighting the therapeutic promise of C004019 PROTAC for AD and tauopathies.² This study signals a new era in drug development targeting tau pathology, offering hope for effective treatments in neurodegenerative diseases.

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