Structure-Based Optimization of Covalent, Small-Molecule Stabilizers of the 14-3-3σ/ERα Protein-Protein Interaction from Nonselective Fragments.
Structure-Based Optimization of Covalent, Small-Molecule Stabilizers of the 14-3-3σ/ERα Protein-Protein Interaction from Nonselective Fragments.
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DOI：
10.1021/jacs.3c05161
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发表时间：
2023-09-20
影响因子：
15
通讯作者：
Arkin, Michelle R.
中科院分区：
文献类型：
--
作者：
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.
蛋白质 - 蛋白质相互作用（PPIs）的稳定化已成为化学生物学和药物研发中一种有前景的策略。确定用于稳定天然PPIs的合适起点以及随后将其精心设计成具有选择性和强效的分子胶水，缺乏基于结构的优化策略。我们先前已经鉴定出一种二硫键片段，它能稳定枢纽蛋白14 - 3 - 3σ与其若干底物（包括ERα和C - RAF）的结合。在此，我们展示了基于结构对非选择性片段进行优化，以获得14 - 3 - 3σ/ERα复合物的选择性和高效小分子稳定剂。例如，经过更精心设计的分子胶水在化合物浓度高达150 μM时，对14 - 3 - 3σ/C - RAF没有稳定作用。采用包括质谱和荧光各向异性在内的正交生物物理分析方法来建立构效关系。通过X射线晶体学阐明了37种化合物的结合模式，这进一步辅助了伴随的基于结构的优化。通过靶向14 - 3 - 3σ/ERα界面中的特定氨基酸并用螺环锁定构象，优化后的共价稳定剂181在效力、协同性和选择性方面达到了与天然产物壳梭孢菌素 - A相似的水平。这个案例研究展示了在分子胶水开发中考虑结构、动力学和协同性的价值。
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.
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影响因子：
3.4
作者：
Hallenbeck KK;Turner DM;Renslo AR;Arkin MR
通讯作者：
Arkin MR
影响因子：
18.2
作者：
Kenanova, Dyana N.;Visser, Emira J.;Virta, Johanna M.;Sijbesma, Eline;Centorrino, Federica;Vickery, Holly R.;Zhong, Mengqi;Neitz, R. Jeffrey;Brunsveld, Luc;Ottmann, Christian;Arkin, Michelle R.
通讯作者：
Arkin, Michelle R.
影响因子：
7.3
作者：
Freeman, Alyson K.;Morrison, Deborah K.
通讯作者：
Morrison, Deborah K.
影响因子：
7.3
作者：
Gehringer, Matthias;Laufer, Stefan A.
通讯作者：
Laufer, Stefan A.
影响因子：
120.1
作者：
Boike, Lydia;Henning, Nathaniel J.;Nomura, Daniel K.
通讯作者：
Nomura, Daniel K.