Exploiting Metabolic Vulnerabilities in Cancer with a Dual-Transporter-Targeted 2-Deoxyglucose Analogue for Low-Dose,...

Clinical translation of 2-deoxyglucose (2DG) as a cancer therapeutic has been precluded by the need for high systemic concentrations to outcompete circulating glucose, resulting in dose-limiting toxicity. Here, we overcome this barrier by covalently linking 2DG to 1,18-octadecanedioic acid (ODDA) to engage both glucose and long chain fatty acid transport pathways simultaneously. The resulting conjugate (2DG-ODDA) associates with serum albumin and leverages dual-transporter uptake by 4T1 triple-negative breast cancer (TNBC) cells. In vitro, 2DG-ODDA is highly potent as compared to the parent 2DG, with a 16-fold lower IC₅₀ value. Analysis of how 2DG-ODDA exerts its cytotoxic effects revealed that the compound induces apoptosis consistent with glycolytic inhibition, as confirmed by metabolic flux analysis showing a significant decrease in glycolysis-derived ATP production. 2DG-ODDA treatment also significantly reduces the rate of mitochondrial ATP production by cells, indicating that the conjugate disrupts multiple cellular processes in its mechanism of action. Further analysis revealed that 2DG-ODDA is cleaved by α-mannosidases. In vivo, low-dose subcutaneous 2DG-ODDA treatment significantly suppresses tumor growth, whereas equimolar 2DG is inactive. Together, these findings establish dual nutrient-pathway targeting as a strategy to confer anticancer activity to otherwise ineffective glycolytic inhibitors by exploiting metabolic vulnerabilities in cancer.