Design, synthesis and functional characteristics of low molecular weight proline-rich motif (PRM) mimetics recognized by PRM binding domains

小分子难以调节蛋白质-蛋白质相互作用（PPI）的一个主要原因是它们通常具有平坦且相当大的结合界面，显示出很少的凹度。这些相互作用根据其特异性进行分类。一类重要的蛋白质结构域显示出溶剂暴露区域，能够结合其靶蛋白的富含脯氨酸的基序（PRM）。目前，已知6个不同的prm结合域（PRO） (Src-homology 3 （SH3）域、WW域、EVH1 （Ena/VASP homology 1）域、GYF域、UEV （ubiquitin E2 variant）域和profilin。这种prd存在于许多多组分信号复合物中，通过在其表面堆叠芳香氨基酸来识别脯氨酸残基的特征模式。此外，这种相互作用的特异性受到侧翼结合表位的影响。这个项目的目的是发现人类EVH1结构域、src激酶Fyn的sh3结构域和YAP-WW结构域的新的特异性调节剂，基于一个新的聚脯氨酸II螺旋模拟结构和先前项目阶段开发和表征的结构修饰。这些化合物代表了一类新的PRM模拟物，可能为开发特异性的、药理学相关的PRM介导的蛋白-蛋白相互作用（PPI）抑制剂提供äs起点。此外，我们通过筛选ChemBioNet文库和虚拟筛选特异性组装的重点文库来寻找新的先导化合物来调节三个结构域家族。本项目结合合理的、基于结构的小分子设计、结构生物学、新型支架的立体选择合成、多肽合成、基于支架的新型连接策略、PRO调节剂的化合物筛选和功能表征。

A major reason for the difficulty in modulating protein-protein interactions (PPI) by small molecules is that they typically possess flat and rather large binding interfaces, displaying little concavity. These interactions are classified according to their specificity. An important dass of such protein domains exhibits solvent exposed regions that are able to bind the proline-rich motifs (PRM) of their target proteins. Currently, six distinct families of PRM-binding domains (PRO) (Src-homology 3 (SH3) domains, WW domains, EVH1 (Ena/VASP homology 1) domains, GYF domains, UEV (ubiquitin E2 variant) domains and profilin are known. Present in many multi-component signaling complexes, such PRDs recognize characteristic patterns of proline residues by means of stacked aromatic amino acids on their surface. Additionally, the specificity of this interaction is influenced by flanking binding epitopes. The aim of this project is to discover new specific modulators of the human EVH1 domain, the SH3-domain of the src kinase Fyn, and the YAP-WW domain, based on a new polyproline II helix mimetic structure and structural modifications developed and characterized during the previous project phase. These compounds represent a new dass of PRM mimetics and may serve äs starting points for the development of specific, pharmacologically relevant inhibitors of PRM- mediated protein-protein interaction (PPI). Additionally, we search for new lead compounds to modulate the three families of domain by screening the ChemBioNet library and by virtual screening of specifically assembled focused libraries. The project combines rational, structurebased small molecule design, structural biology, stereoselective synthesis of novel scaffolds, peptide synthesis, new scaffold-based ligation strategies, compound screening and functional characterization of the PRO modulators.