Faucher Franco F, Abegg Daniel, Ipock Phillip, Adibekian Alexander, Lovell Scott, Bogyo Matthew
Department of Chemistry, Stanford University.
Department of Chemistry, University of Illinois Chicago, Chicago, Illinois 60607, USA.
bioRxiv. 2023 Feb 18:2023.02.17.529022. doi: 10.1101/2023.02.17.529022.
Macrocyclic peptides are attractive for chemoproteomic applications due to their modular synthesis and potential for high target selectivity. We describe a solid phase synthesis method for the efficient generation of libraries of small macrocycles that contain an electrophile and alkyne handle. The modular synthesis produces libraries that can be directly screened using simple SDS-PAGE readouts and then optimal lead molecules applied to proteomic analysis. We generated a library of 480 macrocyclic peptides containing the weakly reactive fluorosulfate (OSF) electrophile. Initial screening of a subset of the library containing each of the various diversity elements identified initial molecules of interest. The corresponding positional and confirmational isomers were then screened to select molecules that showed specific protein labeling patterns that were dependent on the probe structure. The most promising hits were applied to standard chemoproteomic workflows to identify protein targets. Our results demonstrate the feasibility of rapid, on-resin synthesis of diverse macrocyclic electrophiles to generate new classes of covalent ligands.
大环肽因其模块化合成以及具有高靶向选择性的潜力，在化学蛋白质组学应用中颇具吸引力。我们描述了一种固相合成方法，用于高效生成包含亲电试剂和炔烃手柄的小大环文库。这种模块化合成产生的文库可使用简单的SDS-PAGE读数直接进行筛选，然后将最佳先导分子应用于蛋白质组学分析。我们生成了一个包含弱反应性氟代硫酸酯（OSF）亲电试剂的480种大环肽文库。对包含各种不同多样性元素的文库子集进行初步筛选，确定了最初感兴趣的分子。然后对相应的位置异构体和构象异构体进行筛选，以选择显示出依赖于探针结构的特定蛋白质标记模式的分子。最有前景的命中物被应用于标准的化学蛋白质组学工作流程，以鉴定蛋白质靶点。我们的结果证明了快速在树脂上合成各种大环亲电试剂以生成新型共价配体的可行性。
