The dynamic field of cancer research is continually evolving, driven by the pursuit of deeper understanding and novel therapeutic strategies against this multifaceted disease. Metabolomics, an emerging discipline, offers a wide-angle view of the metabolic shifts occurring in organisms. This article explores the application of metabolomics in preclinical cancer research, focusing on its critical role in the discovery of biomarkers, the assessment of therapeutic efficacy and safety, and the facilitation of drug-target mapping within the scope of immuno-oncology.