NAT-A exhibits anti-tumor activity against acquired epidermal growth factor receptor- tyrosine kinase inhibitor-resistant non-small cell lung cancer via inhibiting the β-catenin/PKM2 axis.

Hyperactivation of the Wnt/β-catenin pathway is a key driver of epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) resistance in non-small cell lung cancer (NSCLC), with pyruvate kinase M2 (PKM2) serving as a critical transcriptional coactivator of β-catenin. We show that PKM2 upregulation is associated with TKI resistance through stabilizing β-catenin and amplifying Wnt/β-catenin signaling, suggesting a non-canonical or catalytic activity-independent role of PKM2. Here, we identify Neoantimycin-A (NAT-A), a natural compound isolated from Streptomyces conglobatus by our group, as a novel therapeutic agent that suppresses EGFR-TKI-resistant NSCLC. Mechanistically, NAT-A disrupts the interaction between phosphorylated PKM2 (Ser37/Y105) and β-catenin, promoting β-catenin degradation in a PKM2-dependent manner and subsequent suppression of downstream oncogenic signaling. Both in vitro and in vivo studies confirm NAT-A's anti-tumor efficacy, including inhibition of cell proliferation and induction of apoptosis in EGFR-TKI-resistant models. Our findings demonstrate that activation of the PKM2/β-catenin axis drives EGFR-TKI resistance, and highlight NAT-A as a candidate agent that interferes with their interaction to exert anti-tumor effects against refractory EGFR-TKI-resistant NSCLC.