Noncovalent inhibitors of p97 have entered clinical studies. Compared with noncovalent inhibitors, covalent inhibitors have unique advantages in maintaining inhibitory effect and improving the resistance of the target. We previously employed the activity-based protein profiling to definitely identify p97 as the protein target of FL-18 that has a unique scaffold of benpropargylamide coupled with an imidazole. In this study, we report a thorough structure-activity-relationship study involving the new scaffold. A total of three rounds of optimization led to the discovery of the most potent covalent inhibitor of p97 to date. A chemical proteomics study indicated that the newly-synthesized compounds still targeted the C522 residue of p97 and retained selectivity among the complicated whole proteome. This study provides a suite of new covalent inhibitors of p97 to assist in its biological study and drug discovery.

p97的非共价抑制剂已进入临床研究。与非共价抑制剂相比，共价抑制剂在维持抑制效果、提高靶点耐药性方面具有独特的优势。我们之前使用基于活性的蛋白质分析来确定 p97 作为 FL-18 的蛋白质靶标，FL-18 具有独特的苯丙酰胺与咪唑偶联的支架。在这项研究中，我们报告了涉及新支架的彻底的结构-活性-关系研究。总共三轮优化发现了迄今为止最有效的 p97 共价抑制剂。化学蛋白质组学研究表明，新合成的化合物仍然针对 p97 的 C522 残基，并在复杂的整个蛋白质组中保留了选择性。这项研究提供了一套新的 p97 共价抑制剂，以协助其生物学研究和药物发现。