As one of the well-known hallmarks of cancer malignancy, most proliferating cancer cells exhibit enhanced rates of glycolysis. Hexokinase 2 (HK2) is the rate-limiting enzyme catalyzing the first step of glycolysis, and is often overexpressed in most cancer cells. Thus, targeting HK2 appears to be a promising anticancer therapy. However, selective inhibition of HK2 and the polar nature of the target site remain challenges to the development of small-molecule inhibitors, which could be addressed by targeting unique domains of HK2, such as its N-terminal domain. Here, we review different target–inhibitor binding modes and the associated pharmacological effects, which would be informative for rational molecular design. We also highlight further perspectives and strategies to develop novel HK2 inhibitors for cancer therapy.