PII-027 - CHARACTERIZING NETUPITANT'S FUNCTION AS AN NADK2 INHIBITOR: IMPACT ON CELL VIABILITY AND METABOLISM.

E. Abor¹, J. Yu¹, E. Zhang¹; ¹University of Cincinnati, Cincinnati, OH, USA.

Background: Cancer is a major global health concern, driving the need for new treatment options. Recent advancements in computational drug discovery and proteomics have revealed potential drug candidates and their mechanisms. NADK2 (NAD kinase 2) is a promising target associated with cellular metabolism and various cancers. Netupitant, initially developed antiemetic for chemotherapy-induced nausea, functions by antagonizing neurokinin-1 (NK1) receptors. This study utilized computational tools, proteomic and cellular analyses to investigate netupitant as an NADK2 inhibitor, supporting its further exploration as a novel cancer treatment.

Methods: We investigated netupitant as an NADK2 inhibitor using computational and experimental methods. Molecular docking simulations with AutoDock Vina assessed netupitant's binding to NADK2 based on its crystal structure. Human cancer cell lines (MDA-MB-468, TSC2 null LAM cells) were cultured and treated with varying concentrations of netupitant for up to 72 hours. Cell viability was measured using the crystal violet assay. Proteomic analysis via western blots identified differentially expressed proteins . Cell death and vacuole formations were evaluated using immunofluorescence and confocal imaging. Statistical significance was determined by ANOVA (p < 0.05).

Results: Molecular docking simulations showed that netupitant effectively binds to the active site of NADK2, with a strong binding affinity of -10.9 kcal/mol, suggesting it may inhibit NADK2 activity. Experiments with TSC2 null LAM and MDA-MB-468 cells demonstrated that netupitant significantly reduced cell viability in a dose- and time-dependent manner (p < 0.01). Western blot analysis indicated decreased NADK2 levels and increased pro-apoptotic markers like cleaved caspase-3. Immunofluorescence confirmed enhanced cell death and vacuole formation, supporting netupitant’s role as an NADK2 inhibitor that promotes cell death in cancer cells.

Conclusion: For the first time, this study shows that netupitant is an effective NADK2 inhibitor with potential as a novel cancer treatment. Molecular docking, cell viability assays, and proteomic analyses demonstrate it's ability to disrupt cancer cell metabolism and promote apoptosis, especially in TSC2 null LAM and MDA-MB-468 cells.