PFK-015 (PFK15) is an effective inhibitor of PFKFB3 (IC50: 110 nM) and inhibits PFKFB3 activity in Y cells (IC50: 20 nM).

PFKFB3:110 nM, PFKFB3:20 nM

PFK-015 (25 mg/kg, i.p.) inhibits the growth, metastatic spread, and glucose metabolism of LLC tumors in syngeneic mice. In three distinct athymic mouse models of xenografted human cancers, the anticancer efficacy of PFK-015 is comparable to that of established chemotherapy drugs. PFK-015 exhibits favorable pharmacokinetic properties in vivo.

PFK-015 effectively reduces levels of F26BP, glucose uptake, and intracellular adenosine triphosphate (ATP) in Jurkat T-cell leukemia cells and H522 lung adenocarcinoma cells. Additionally, PFK-015 exhibits significant inhibitory effects on the growth of a variety of cancer cells.

Recombinant PFKFB3 assay: Kinase reactions are conducted by incubating 13 ng of recombinant human PFKFB3 protein in a reaction mix containing 10 μmol/L ATP, 10 μmol/L F6P, and either dimethyl sulfoxide (DMSO) vehicle control, 3PO, or PFK15 for 1 hour at room temperature. Kinase activity is measured with the Adapta Universal Kinase Assay per manufacturer

Viability is determined using trypan blue exclusion. Cells were incubated in 20% trypan blue for 5 minutes. Cells excluding trypan blue are counted using a standard hemocytometer to determine the total number of viable cells. Experiments are conducted in triplicate. (Only for Reference)