Protein Degradation by In-Cell Self-Assembly of Proteolysis Targeting Chimeras
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High Resolution Image Download MS PowerPoint Slide Selective degradation of proteins by proteolysis targeting chimeras (PROTACs) offers a promising potential alternative to protein inhibition for therapeutic intervention. Current PROTAC molecules incorporate a ligand for the target protein, a linker, and an E3 ubiquitin ligase recruiting group, which bring together target protein and ubiquitinating machinery. Such hetero-bifunctional molecules require significant linker optimization and possess high molecular weight, which can limit cellular permeation, solubility, and other drug-like properties. We show here that the hetero-bifunctional molecule can be formed intracellularly by bio-orthogonal click combination of two smaller precursors. We designed a tetrazine tagged thalidomide derivative which reacts rapidly with a trans -cyclo-octene tagged ligand of the target protein in cells to form a cereblon E3 ligase recruiting PROTAC molecule. The in-cell click-formed proteolysis targeting chimeras (CLIPTACs) were successfully used to degrade two key oncology targets, BRD4 and ERK1/2. ERK1/2 degradation was achieved using a CLIPTAC based on a covalent inhibitor. We expect this approach to be readily extendable to other inhibitor-protein systems because the tagged E3 ligase recruiter is capable of undergoing the click reaction with a suitably tagged ligand of any protein of interest to elicit its degradation.
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Modification of the E3 ligands prevents the recruitment of their respective E3 ligases by the PROTAC, allowing assessment of PROTAC activity in the absence of the E3 ligase.
“The modification of the E3 ligands prevents the recruitment of their respective E3 ligases by the PROTAC, thereby allowing the assessment of the activity of the PROTAC in the absence of the E3 ligase45,46.”
From An HDAC4-specific PROTAC degrader achieves radiation sensitization by enhancing ferroptosis in lung cancer (2026) · cites this paper as [45]
The Introduction describes methylated Tz-thalidomide as a control that disrupts binding to CRBN, and the Results (BRD4 Degradation by CLIPTAC) report no BRD4 degradation with that control. This supports testing activity without effective CRBN recruitment, but the paper does not establish the claim for E3 ligands generally or test activity in cells physically lacking the E3 ligase. (LLM, conf=0.84)
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- Authors
- Lebraud, H., Wright, D. J., Johnson, C. N., & Heightman, T. D.
- Affiliations
- Astex Pharmaceuticals, 436 Cambridge Science Park, Cambridge, CB4 0QA, U.K.
- Corresponding author
- Heightman, Tom D.
- Venue
- ACS Central Science
- Publisher
- American Chemical Society (ACS)
- Published
- 2016-12-05
- Volume
- 2
- Issue
- 12
- Pages
- 927-934
- ISSN
- 2374-7943 (ISSN-L)
- 2374-7951
- PMC
- PMC5200928
- OpenAlex
- W2559947248
- Cited by
- 413
- References
- 32
- Citations by year
- 2026: 46 · 2025: 59 · 2024: 54 · 2023: 56 · 2022: 52 · 2021: 39 · 2020: 37 · 2019: 33 · 2018: 18 · 2017: 18 · 2014: 1
- FWCI
- 12.13
- Type
- journal article
- Language
- en
- Open access
- diamond
- doi.org
- Licence
- Locations
- ACS Central Science (publishedVersion, other-oa)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Degradation and Inhibitors
- Physical Sciences › Chemistry › Organic Chemistry › Click Chemistry and Applications
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ubiquitin and proteasome pathways
- Keywords
- PROTACs, bioorthogonal click chemistry, thalidomide derivatives, trans-cyclooctene, BRD4 degradation, covalent inhibitors, protein degradation, heterobifunctional molecules, linker optimization
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