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Structure-Based Optimization of Covalent, Small-Molecule Stabilizers of the 14-3-3σ/ERα Protein–Protein Interaction from Nonselective Fragments
[Image: see text] The stabilization of protein–protein interactions (PPIs) has emerged as a promising strategy in chemical biology and drug discovery. The identification of suitable starting points for stabilizing native PPIs and their subsequent elaboration into selective and potent molecular glues...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical Society
2023
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10515640/ https://www.ncbi.nlm.nih.gov/pubmed/37676236 http://dx.doi.org/10.1021/jacs.3c05161 |
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author | Konstantinidou, Markella Visser, Emira J. Vandenboorn, Edmee Chen, Sheng Jaishankar, Priyadarshini Overmans, Maurits Dutta, Shubhankar Neitz, R. Jeffrey Renslo, Adam R. Ottmann, Christian Brunsveld, Luc Arkin, Michelle R. |
author_facet | Konstantinidou, Markella Visser, Emira J. Vandenboorn, Edmee Chen, Sheng Jaishankar, Priyadarshini Overmans, Maurits Dutta, Shubhankar Neitz, R. Jeffrey Renslo, Adam R. Ottmann, Christian Brunsveld, Luc Arkin, Michelle R. |
author_sort | Konstantinidou, Markella |
collection | PubMed |
description | [Image: see text] The stabilization of protein–protein interactions (PPIs) has emerged as a promising strategy in chemical biology and drug discovery. The identification of suitable starting points for stabilizing native PPIs and their subsequent elaboration into selective and potent molecular glues lacks structure-guided optimization strategies. We have previously identified a disulfide fragment that stabilized the hub protein 14-3-3σ bound to several of its clients, including ERα and C-RAF. Here, we show the structure-based optimization of the nonselective fragment toward selective and highly potent small-molecule stabilizers of the 14-3-3σ/ERα complex. The more elaborated molecular glues, for example, show no stabilization of 14-3-3σ/C-RAF up to 150 μM compound. Orthogonal biophysical assays, including mass spectrometry and fluorescence anisotropy, were used to establish structure–activity relationships. The binding modes of 37 compounds were elucidated with X-ray crystallography, which further assisted the concomitant structure-guided optimization. By targeting specific amino acids in the 14-3-3σ/ERα interface and locking the conformation with a spirocycle, the optimized covalent stabilizer 181 achieved potency, cooperativity, and selectivity similar to the natural product Fusicoccin-A. This case study showcases the value of addressing the structure, kinetics, and cooperativity for molecular glue development. |
format | Online Article Text |
id | pubmed-10515640 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | American Chemical Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-105156402023-09-23 Structure-Based Optimization of Covalent, Small-Molecule Stabilizers of the 14-3-3σ/ERα Protein–Protein Interaction from Nonselective Fragments Konstantinidou, Markella Visser, Emira J. Vandenboorn, Edmee Chen, Sheng Jaishankar, Priyadarshini Overmans, Maurits Dutta, Shubhankar Neitz, R. Jeffrey Renslo, Adam R. Ottmann, Christian Brunsveld, Luc Arkin, Michelle R. J Am Chem Soc [Image: see text] The stabilization of protein–protein interactions (PPIs) has emerged as a promising strategy in chemical biology and drug discovery. The identification of suitable starting points for stabilizing native PPIs and their subsequent elaboration into selective and potent molecular glues lacks structure-guided optimization strategies. We have previously identified a disulfide fragment that stabilized the hub protein 14-3-3σ bound to several of its clients, including ERα and C-RAF. Here, we show the structure-based optimization of the nonselective fragment toward selective and highly potent small-molecule stabilizers of the 14-3-3σ/ERα complex. The more elaborated molecular glues, for example, show no stabilization of 14-3-3σ/C-RAF up to 150 μM compound. Orthogonal biophysical assays, including mass spectrometry and fluorescence anisotropy, were used to establish structure–activity relationships. The binding modes of 37 compounds were elucidated with X-ray crystallography, which further assisted the concomitant structure-guided optimization. By targeting specific amino acids in the 14-3-3σ/ERα interface and locking the conformation with a spirocycle, the optimized covalent stabilizer 181 achieved potency, cooperativity, and selectivity similar to the natural product Fusicoccin-A. This case study showcases the value of addressing the structure, kinetics, and cooperativity for molecular glue development. American Chemical Society 2023-09-07 /pmc/articles/PMC10515640/ /pubmed/37676236 http://dx.doi.org/10.1021/jacs.3c05161 Text en © 2023 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by/4.0/Permits the broadest form of re-use including for commercial purposes, provided that author attribution and integrity are maintained (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Konstantinidou, Markella Visser, Emira J. Vandenboorn, Edmee Chen, Sheng Jaishankar, Priyadarshini Overmans, Maurits Dutta, Shubhankar Neitz, R. Jeffrey Renslo, Adam R. Ottmann, Christian Brunsveld, Luc Arkin, Michelle R. Structure-Based Optimization of Covalent, Small-Molecule Stabilizers of the 14-3-3σ/ERα Protein–Protein Interaction from Nonselective Fragments |
title | Structure-Based Optimization
of Covalent, Small-Molecule
Stabilizers of the 14-3-3σ/ERα Protein–Protein
Interaction from Nonselective Fragments |
title_full | Structure-Based Optimization
of Covalent, Small-Molecule
Stabilizers of the 14-3-3σ/ERα Protein–Protein
Interaction from Nonselective Fragments |
title_fullStr | Structure-Based Optimization
of Covalent, Small-Molecule
Stabilizers of the 14-3-3σ/ERα Protein–Protein
Interaction from Nonselective Fragments |
title_full_unstemmed | Structure-Based Optimization
of Covalent, Small-Molecule
Stabilizers of the 14-3-3σ/ERα Protein–Protein
Interaction from Nonselective Fragments |
title_short | Structure-Based Optimization
of Covalent, Small-Molecule
Stabilizers of the 14-3-3σ/ERα Protein–Protein
Interaction from Nonselective Fragments |
title_sort | structure-based optimization
of covalent, small-molecule
stabilizers of the 14-3-3σ/erα protein–protein
interaction from nonselective fragments |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10515640/ https://www.ncbi.nlm.nih.gov/pubmed/37676236 http://dx.doi.org/10.1021/jacs.3c05161 |
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