Notch激活复合物激酶（NACK）是Notch转录机制的一个组成部分，对于Notch介导的肿瘤形成至关重要。然而，NACK调节Notch介导的转录的机制尚不明确。在这里，我们证明NACK结合并水解ATP，只有ATP结合的NACK才能结合于Notch三元复合物（NTC）。基于此，我们试图寻找这种ATP依赖性功能的抑制剂，并使用计算流程发现了第一个小分子抑制剂Z271-0326，它直接阻断了Notch介导的转录活性，并在PDX小鼠模型中表现出强大的抗肿瘤活性。总之，我们发现了第一个抑制剂，可以通过阻断NTC下游的Notch活性，为Notch驱动的癌症的有效治疗提供希望。 © 2023作者。

Notch activation complex kinase (NACK) is a component of the Notch transcriptional machinery critical for the Notch-mediated tumorigenesis. However, the mechanism through which NACK regulates Notch-mediated transcription is not well understood. Here, we demonstrate that NACK binds and hydrolyzes ATP and that only ATP-bound NACK can bind to the Notch ternary complex (NTC). Considering this, we sought to identify inhibitors of this ATP-dependent function and, using computational pipelines, discovered the first small-molecule inhibitor of NACK, Z271-0326, that directly blocks the activity of Notch-mediated transcription and shows potent antineoplastic activity in PDX mouse models. In conclusion, we have discovered the first inhibitor that holds promise for the efficacious treatment of Notch-driven cancers by blocking the Notch activity downstream of the NTC.© 2023 The Author(s).