OC201 and OC202e, an AI-driven drug combination, enhance the susceptibility of pancreatic cancer to FOLFIRINOX by modulating HIF-1alpha

Objective: Pancreatic cancer remains highly lethal, with a five-year survival rate of ~13%, largely due to late-stage diagnosis, aggressive progression, and resistance to standard therapies. Although FOLFIRINOX is the current first-line regimen, therapeutic resistance limits its durability. Our previous work showed that the AI-derived combination OC201 and OC202e suppresses EMT and metastasis. This study investigated whether OC201/OC202e enhances the response to FOLFIRINOX by modulating multidrug-resistance mechanisms. Methods: Panc-1 and MIA PaCa-2 cells were treated with OC201 and/or OC202e followed by FOLFIRINOX. Cell viability and clonogenic potential were assessed using MTT and colony-formation assays. Multidrug-resistance determinants-including hypoxia, extracellular matrix factors, dysregulated signaling, genetic alterations, stemness, and ABC transporters were analyzed by Western blotting and RT-qPCR. Stemness was further evaluated using sphere-formation assays. An in vivo hepatic metastasis model was established via intrasplenic injection of Panc-1 cells, followed by 10 weeks of treatment with OC201, OC202e, and/or FOLFIRINOX beginning two weeks post-injection. Metastatic burden was quantified histologically. Results: OC201/OC202e significantly enhanced FOLFIRINOX efficacy, as shown by reduced colony formation in both cell lines, while MTT assays indicated limited short-term synergy, suggesting a primarily long-term effect. In vivo, the combination further reduced hepatic metastatic lesions compared with FOLFIRINOX alone. Mechanistic analyses revealed decreased HIF-1alpha and stemness markers (CD44, Flotillin2) following combination treatment. HIF-1alpha expression correlated with stemness and FOLFIRINOX responsiveness, supporting its role in resistance. These findings align with the known EMT-stemness link and provide mechanistic insight into the observed synergy. Conclusion: The AI-derived OC201/OC202e combination enhances sensitivity to FOLFIRINOX and suppresses metastasis by targeting HIF-1alpha and the EMT-stemness axis. Reductions in clonogenicity and resistance-related markers highlight its potential as an adjuvant to standard chemotherapy and underscore the promise of AI-guided strategies for developing synergistic therapeutic combinations.