Repurposed a mixture of Canagliflozin and metronidazole as Novel Dual-Target Therapy for Cervical Cancer: Mechanistic Insights into targeting the Akt pathway

Objective: This study explores repurposing combined Canagliflozin and Metronidazole therapy as a dual-target approach for cervical cancer, focusing on Akt signaling mechanisms.Materials and methods: This study systematically evaluated the cytotoxic effects of Canagliflozin, Metronidazole, their 1:1 combination, and the reference drug 5-Fluorouracil (5-FU) on HeLa (human cervical cancer) and HFF (normal human Foreskin Fibroblast) cell lines. Using the MTT assay at 24 and 72 hours, cellular viability was assessed, and the selective toxicity index (SI) was calculated to determine specificity towards cancer cells. Drug interactions were analyzed through combination index (CI) using Compusyn software. Molecular docking simulation predicted compounds' binding affinities to kinases in the Akt pathway, including PI3K, PDK1, Akt, and mTOR.Results: The drug combination demonstrated potent, selective cytotoxicity against HeLa cells, with an IC₅₀ of 25.1 µg/mL at 72 hours, while showing minimal toxicity to HFF cells (IC₅₀ > 1000 µg/mL). The selectivity index (SI) surpassed that of 5-fluorouracil, and the combination index (CI) analysis indicated strong synergy. Molecular docking revealed substantial binding affinities to the Akt pathway, with docking scores of -10.5, -9.0, -9.7, and -8.9 kcal/mol for canagliflozin and -5.5, -5.4, -5.5, and -5.1 kcal/mol for metronidazole across PI3K, PDK1, Akt, and mTOR kinases. Conclusion: The mixture demonstrates notable synergistic anticancer activity with a high degree of selectivity for cervical cancer cells. This effect is likely mediated through the suggested dual inhibition of the Akt signaling pathway. Such a repurposing strategy presents a promising targeted therapeutic approach that warrants further investigation.