In silico screening and in vitro biological evaluation reveal Queuine as a promising MAP4K4 inhibitor for treating pancreatic cancer.

Pancreatic cancer (PC) remains one of the deadliest cancer types, with limited success in treatment despite advances in research. MAP4K4 is overexpressed in pancreatic tumors and linked to disease progression, making it a promising target for PC therapy. This study aimed to identify bioactive nutraceutical molecules targeting MAP4K4 using molecular docking, molecular dynamics (MD) simulations, and MM-GBSA calculations. The computational findings were validated through in vitro MTT cell viability assays and MAP4K4 enzyme ELISA tests. Queuine and Thiamine were identified as potent MAP4K4 inhibitors with comparable docking scores to the approved drug Gemcitabine. MD simulations confirmed stable binding for 100 ns (3 runs), with average binding free energy values of -50 kcal/mol for Queuine, -47 kcal/mol for Thiamine, and − 18 kcal/mol for Gemcitabine. In vitro assays showed that Thiamine was non-cytotoxic at high concentrations, while Queuine had a significantly lower IC50 (5.95 µM) compared to Gemcitabine (64.17 µM), making it nearly ten times more potent. MAP4K4 ELISA tests confirmed Queuine’s superior binding and enzyme inhibition compared to Gemcitabine. Synergy studies combining Queuine (0.25–1.25 µM) and Gemcitabine (0.05–2.5 µM) revealed strong synergistic effects, suggesting enhanced efficacy at lower doses. These findings highlight Queuine as a promising natural therapeutic agent for PC, with potential for combination therapy with Gemcitabine. Further in vivo studies are recommended to explore its therapeutic potential.