Development of Novel Phosphonodifluoromethyl-Containing Phosphotyrosine Mimetics and a First-In-Class, Potent, Selective, and Bioavailable Inhibitor of Human CDC14 Phosphatases

Together with protein tyrosine kinases, protein tyrosine phosphatases (PTPs) control protein tyrosine phosphorylation and regulate numerous cellular functions. Dysregulated PTP activity is associated with the onset of multiple human diseases. Nevertheless, understanding of the physiological function and disease biology of most PTPs remains limited, largely due to the lack of PTP-specific chemical probes. In this study, starting from a well-known nonhydrolyzable phosphotyrosine (pTyr) mimetic, phosphonodifluoromethyl phenylalanine (F2Pmp), we synthesized 7 novel phosphonodifluoromethyl-containing bicyclic/tricyclic aryl derivatives with improved cell permeability and potency toward various PTPs. Furthermore, with fragment- and structure-based design strategies, we advanced compound 9 to compound 15, a first-in-class, potent, selective, and bioavailable inhibitor of human CDC14A and B phosphatases. This study demonstrates the applicability of the fragment-based design strategy in creating potent, selective, and bioavailable PTP inhibitors and provides a valuable probe for interrogating the biological roles of hCDC14 phosphatases and assessing their potential for therapeutic interventions.

蛋白酪氨酸激酶（protein tyrosine kinases）与蛋白酪氨酸磷酸酶（protein tyrosine phosphatases, PTPs）协同调控蛋白质酪氨酸磷酸化进程，并调节诸多细胞生理功能。PTP活性异常失调与多种人类疾病的发生发展密切相关。然而，由于缺乏PTP特异性化学探针，目前学界对绝大多数PTP的生理功能及疾病相关生物学机制的认知仍存在显著局限。本研究以经典的非水解型磷酸酪氨酸（phosphotyrosine, pTyr）模拟物——膦酰二氟甲基苯丙氨酸（phosphonodifluoromethyl phenylalanine, F2Pmp）为起始骨架，合成了7种全新的含膦酰二氟甲基双环/三环芳基衍生物，这类衍生物对多种PTP展现出更优异的细胞通透性与抑制效力。此外，本研究依托基于片段与结构的联合设计策略，将化合物9优化升级为化合物15——一款同类首款、强效且具有高选择性的人源CDC14A与CDC14B磷酸酶抑制剂，同时具备良好的生物可利用性。本研究证实了基于片段的设计策略在开发强效、高选择性且生物可利用的PTP抑制剂中的适用性，同时为探究hCDC14磷酸酶的生物学功能、评估其在治疗干预中的应用潜力提供了极具价值的化学探针。