Phosphofructokinase-1 (PFK-1) is a key enzyme in glycolysis that recommences the transformation of fructose-6-phosphate into fructose-1,6-bisphosphate, likely a rate control point of energy metabolism. PKF-1 is involved in the cellular energy metabolism process and is regulated both at the molecular level by feedback inhibition and at the post-translational level. The altered regulation of PFK-1 is intrinsic to numerous neoplasms, including cancers in which increased aerobic glycolysis is linked with rapid cell division and tumor resistance. Since PFK-1 is also related to cancer metabolism, especially the increased metabolism, the target point may be of great value. PFK-1 is a potential therapeutic target because it is involved in automating glycolytic flux and, subsequently, is essential for tumor growth and therapy resistance, especially in triple-negative breast cancer. Apart from cancer, PFK-1 is involved in metabolic and inflammation diseases, for example, diabetes, obesity, and cardiovascular disease, because it regulates glucose homeostasis insulin sensitivity, and macrophage polarization. Inhibiting PFK-1 could have potential benefits in these diseases since it suggests that treatment can target system inflammation and energy production. However, new issues arise with the treatment in that creating a selective, low-toxicity PFK-1 inhibitor that takes all of PFK-1 physiological functions into mind can still be regarded as a tricky exertion. This chapter is specifically devoted to discussing the general biochemical and physiological functions of PFK-1, the regulation of this enzyme, and its contribution to pathophysiology. Further design of specific inhibitors and individualized approaches to the treatment of cancer and metabolic disorders will dramatically shift therapeutic paradigms by including PFK-1 as a target. More in-depth studies of the structure, regulation, and role of PFK-1 will help uncover all kinds of possibilities for using it as a multipurpose target for treatments.