Inhibiting Glycolysis and Disrupting the Mitochondrial HK2–VDAC1 Protein–Protein Interaction Using a Bifunctional Lonidamine-Conjugated Metal Probe for Combating Triple-Negative Breast Cancer

Triple-negative breast cancer (TNBC) relies primarily on aerobic glycolysis for energy and rapid cancer cell proliferation. Hexokinase 2 (HK2), a key enzyme regulating glycolysis, is overexpressed in TNBC, promoting tumor cell proliferation and apoptosis resistance by interacting with the mitochondrial membrane's voltage-dependent anion channel 1 (VDAC1). However, the development of bioactive molecules for effectively disrupting the HK2-VDAC1 interaction remains challenging. Herein, we have modified londamine (LND) with an iridium(III) complex to create bifunctional far-red probe <b>1</b>. This complex not only has the ability to distinguish TNBC cells from normal cells by probing HK2 in mitochondria, but also significantly enhances antitumor activity by inhibiting mitochondrial glycolysis and effectively disrupting the HK2-VDAC1 interaction. This led to increased Bax-VDAC1 interaction, opening of the mitochondrial permeability transition pores (MPTPs), and generation of ROS, ultimately leading to mitochondrial dysfunction and enhanced cancer cell apoptosis. Probe <b>1</b> also demonstrated stronger antiproliferative activity than LND alone in a TNBC mouse model by targeting the HK2-VDAC1 interaction without causing overt toxicity. This work showcases the potential of probe <b>1</b> as an effective therapeutic agent for TNBC by inhibiting the mitochondrial HK2-VDAC1 interaction.