Mebendazole impairs the expression and function of enzymes in nucleotide metabolism pathways, leading to Selective Cytotoxicity, Cell Cycle Arrest, and Damage to Cell Morphology in Gastric Cancer.

The nucleotide biosynthetic pathway is essential for tumor replication through DNA and RNA synthesis, making it a critical target for cancer therapies. Mebendazole (MBZ) has shown promise in inhibiting cell proliferation and glucose metabolism in gastric cancer (GC) cell lines. However, its impact on nucleotide synthesis remains unclear. This study investigates MBZ's role in nucleotide biosynthesis and its contribution to antiproliferative effects by modulating key DNA/RNA synthesis targets. MBZ demonstrates antitumor activity similar to 5-FU in GC cells, with lower toxicity to non-tumor cells. Gene expression analysis revealed overexpression of nucleotide metabolism genes (PRPS1, TYMS, MTHFD1, and HPRT1) in tumor samples, and higher levels of TYMS, MTHFD1, and HPRT1 were associated with poor survival in GC patients. MBZ treatment reduced the expression of these genes, induced G0/G1 phase cell cycle arrest, and inhibited metastatic cell proliferation after 48 h. Molecular docking studies indicated that MBZ binds more strongly to PRPS1 and HPRT1 than their natural ligands or inhibitors. Our findings suggest that MBZ modulates nucleotide synthesis pathways, contributing to its selective antiproliferative effects in GC cells. This study highlights potential new pharmacological targets for drug repurposing and further investigation into the broader therapeutic applications of MBZ.