NT-F71, a novel benzothiazole hydrazone, combats azole-resistant candidiasis by inhibiting phosphoglycerate mutase 1 (Gpm1) to suppress glycolysis

<jats:p>&lt;b&gt;Background and Purpose&lt;/b&gt; The global emergence and widespread dissemination of azole-resistant &lt;i&gt;Candida &lt;/i&gt;species create severe challenges for existing antifungal therapies, highlighting an urgent unmet clinical demand for novel antifungal agents and druggable molecular targets to treat candidiasis. Here, we characterised NT-F71, a novel benzothiazole hydrazone derivative, which exerted potent broad-spectrum fungicidal activity against &lt;i&gt;Candida &lt;/i&gt;species, including clinical azole-resistant &lt;i&gt;Candida albicans&lt;/i&gt; isolates. &lt;b&gt;Experimental Approach&lt;/b&gt; Integrated targeted metabolomics, drug affinity responsive target stability (DARTS) and surface plasmon resonance (SPR) assays identified phosphoglycerate mutase 1 (Gpm1) as the direct antifungal target of NT-F71. Subsequent molecular docking and recombinant enzyme activity assays verified that NT-F71 directly binds to and suppresses the catalytic function of &lt;i&gt;C. albicans&lt;/i&gt; Gpm1. &lt;b&gt;Key results&lt;/b&gt; In line with these findings, we found that NT-F71 blocks glycolytic flux via inhibition of Gpm1, which triggers compensatory upregulation of alternative carbon metabolic pathways and disrupts fungal energy homeostasis. &lt;b&gt;Conclusion and Implications&lt;/b&gt; Collectively, our findings demonstrate that targeting Gpm1 with NT-F71 to suppress glycolysis represents a promising antifungal strategy to combat azole-resistant candidiasis.</jats:p>