Accelerated glucose metabolism is a common feature of cancer cells. Hexokinase 2 (HK2) as the rate-limiting enzyme catalyzes the first step of glucose metabolism. It is overexpressed in most of the human cancers and has been a promising target for cancer therapy. Here, we report a novel selective HK2 inhibitor Benitrobenrazide (BNBZ), with nanomolar inhibitory potency. In vitro, BNBZ directly binds to HK2, induces apoptosis, and inhibits proliferation of HK2-overexpressed cancer cells. BNBZ also significantly inhibits the glycolysis of SW1990 cells by targeting HK2. The knockdown or knockout of HK2 expression in SW1990 cells can reduce their sensitivity to BNBZ. Additionally, oral administration of BNBZ can effectively inhibit tumor growth in SW1990 and SW480 xenograft models. In general, BNBZ significantly inhibited glycolysis and cancer cell proliferation in vitro and in vivo by directly targeting HK2 with high potency and low toxicity, and can be developed as a novel HK2 small-molecule candidate drug for future cancer therapeutics.

葡萄糖代谢加速是癌细胞的一个共同特征。己糖激酶 2 (HK2) 作为限速酶催化葡萄糖代谢的第一步。它在大多数人类癌症中过度表达，并且已成为癌症治疗的有希望的靶标。在此，我们报道了一种新型选择性 HK2 抑制剂苯甲酰苯肼 (BNBZ)，具有纳摩尔级抑制效力。在体外，BNBZ 直接与 HK2 结合，诱导细胞凋亡，并抑制 HK2 过表达的癌细胞的增殖。 BNBZ 还通过靶向 HK2 显着抑制 SW1990 细胞的糖酵解。 SW1990细胞中HK2表达的敲低或敲除可以降低其对BNBZ的敏感性。此外，口服BNBZ可以有效抑制SW1990和SW480异种移植模型中的肿瘤生长。总体而言，BNBZ通过直接靶向HK2，在体外和体内显着抑制糖酵解和癌细胞增殖，且高效低毒，可开发为未来癌症治疗的新型HK2小分子候选药物。