Discovery of Novel Glutaminase (GLS1) and Histone Deacetylase (HDAC) Dual Inhibitors as a Promising Strategy for Cancer Therapy.

Glutaminase 1 (GLS1) and histone deacetylases (HDACs) are important targets involved in cancer metabolism and epigenetic regulation, respectively. Herein, we report the design, synthesis, and biological evaluation of first-in-class GLS1/HDAC dual inhibitors as a single-molecule strategy for cancer therapy. Through systematic structure-activity relationship studies and biochemical profiling, compound 58 was identified as a promising dual inhibitor with moderate GLS1 inhibition and potent HDAC activity, particularly against HDAC6. Compound 58 exhibited strong antiproliferative activity in multiple cancer cell lines and modulated GLS1-associated glutamine metabolism and HDAC-related acetylation markers in cells. In a CT26 tumor-bearing mouse model, compound 58 achieved a tumor growth inhibition (TGI) rate of 91.10% without obvious systemic toxicity. Overall, this study supports GLS1/HDAC dual inhibition as a promising anticancer strategy and provides compound 58 as a valuable lead for further optimization.