Dual Repurposing Strategy: Rivaroxaban and Ciprofloxacin as a Synergistic Anticancer Therapy in Cervical Cancer: Insight into Inhibition of Metastasis via Targeting the EMT Pathway

Objective: This study assessed the anticancer and antimetastatic impact of rivaroxaban-ciprofloxacin mixture and explored its molecular mechanism by examining its ability to target the epithelial-mesenchymal transition (EMT) pathway. Methods: Following incubation periods of 24 and 72 hours, HeLa and normal human fibroblast (NHF) cell lines were employed to investigate the anticancer and safety properties of rivaroxaban, ciprofloxacin, rivaroxaban-ciprofloxacin mixture, and carboplatin, with concentrations varying from 0.1 to 1000 µg/ml. The wound healing assay (WHA) of cervical cancer cells was employed to assess the anti-metastatic effects of the mixture at three time points: 0, 12, and 24 hours.. A combination index (CI) and selectivity toxicity index (SI) score were estimated to assess the potential synergistic effects of the mixture’s components and their selectivity toxicity. A computational molecular docking simulation was conducted to evaluate the binding affinity of rivaroxaban and ciprofloxacin to different regulatory signals in the EMT pathway. Results: The MTT and WHS assays outcomes revealed that the rivaroxaban-ciprofloxacin mixture possesses anticancer and antimetastatic properties. This combination significantly suppresses the growth of cervical cancer cells more effectively than carboplatin, rivaroxaban, or ciprofloxacin alone. Additionally, the mixture results in a 98% delay of cancer cell migration after 24 hours of incubation. Furthermore, the mixture’s cytotoxicity on NHF cells was significantly lower than carboplatin. The combination of rivaroxaban and ciprofloxacin showed synergistic cytotoxic effects, as confirmed by the CI score. Also, it displays a selective cytotoxicity toward cancer cells, as indicated by the SI score. The molecular docking study results revealed that the most favorable interactions for rivaroxaban and ciprofloxacin occurred with the matrix metalloprotease and Tankyrase enzyme, with docking scores of -9 kcal/mol and -8.9 kcal/mol, respectively. Conclusion: The study’s findings and the established pharmacokinetic and safety profiles of mixture drugs suggest that the pairing of rivaroxaban and ciprofloxacin offers a promising and safer option for treating cervical cancer.