Discovery of a novel dual-target modulator of ULK1 and ERK1/2 that suppresses triple-negative breast cancer progression and metastasis via the Exo70/Cav-1/MMPs axis.

Triple-negative breast cancer (TNBC) represents an aggressive breast cancer subtype with unfavorable clinical outcomes. Because actionable therapeutic targets remain limited, current treatment options are suboptimal and patient prognosis is generally poor. Our study aims to address the dual challenges of proliferation control and metastasis inhibition in TNBC treatment. Based upon previous findings, although Unc-51 like kinase 1 (ULK1) activators can inhibit proliferation, their anti-metastatic effects may be antagonized by the abnormally activated Extracellular signal-regulated kinase 1/2 (ERK1/2) signaling pathway. Thus, we propose an innovative therapeutic strategy that synergistically targets ULK1 (activation) and ERK1/2 (inhibition). Accordingly, we successfully developed the first ULK1/ERK1/2 dual-target small-molecule modulator, UE01. Kinase activity analysis showed that UE01 can effectively activate ULK1 and inhibit ERK1. Its dual-target binding properties were validated through molecular docking, kinetic simulations, and cellular thermal shift assays (CETSA). The concentration-dependent inhibition of TNBC cell proliferation, colony formation, and migration by UE01 in vitro was superior to or equivalent to single-target positive drugs. In animal models, UE01 significantly inhibits tumor growth and lung metastasis, without showing a remarkable toxicity. The anti-tumor effect of UE01 was achieved on the one hand by inducing autophagic cell death and apoptosis, and on the other hand, its excellent anti-metastatic ability stems from key regulation of the Exo70/Cav-1/MMPs axis. By synergistically activating ULK1 and inhibiting ERK1/2, UE01 could disrupt the functional coupling of Exo70/Cav-1, thereby inhibiting the secretion of MMP-2/MMP-9 and extracellular matrix degradation. Taken together, these results demonstrate that a dual-target strategy against ERK1/2 attenuates TNBC growth and metastatic dissemination by disrupting the Exo70/Cav-1/MMPs axis, highlighting UE01 as a potential lead compound for future targeted therapy.