Benserazide, a dopadecarboxylase inhibitor, suppresses tumor growth by targeting hexokinase 2

Abstract

BackgroundHexokinase (HK) is the rate-limiting enzyme in the first reaction of glycolysis. And Hexokinase 2 (HK2) is most closely related to malignant tumor which expresses at higher level compared with normal cells. HK2 plays a pivotal role in tumor initiation and maintenance, which provides a new target for cancer therapy.MethodsStructure-based virtual ligand screening was used in hit identification from ZINC Drug Database. Microscale thermophoresis assay was performed to evaluate the binding affinity. Enzyme inhibition, cytotoxicity, apoptosis, intracellular ATP level, mitochondrial membrane potential (MMP), glucose uptake and lactate production experiments were undertaken in SW480 cells to identify Benz as a HK2 inhibitor. Western blot was used to test protein expression. SW480 cells xenograft mouse models were used for in vivo study. Nano-particles of Benz were prepared to improve the antitumor efficacy and tumor targeting of Benz. HPLC was used to measure the concentration of free Benz in tumor tissues.ResultsBenserazide (Benz), was identified as a selective HK2 inhibitor, could specifically bind to HK2 and significantly inhibit HK2 enzymatic activity in vitro. In addition, Benz reduced glucose uptake, lactate production and intracellular ATP level, and could cause cell apoptosis and an increased loss of MMP as well. In vivo study indicated that intraperitoneal (ip) injection of Benz at 300 and 600 mg/Kg suppressed cancer growth in tumor-bearing mice and no toxicity shown. To further improve the antitumor efficacy and tumor targeting of Benz, nano-particles of Benz was prepared. Liposomal Benz at 100 and 200 mg/Kg performed potent inhibitory effects on tumor-bearing mice, showing reduced dose and better efficacy.ConclusionsOur study provides a new direction for the development of Benz and its analogues as novel antitumor agents for cancer therapy.Electronic supplementary materialThe online version of this article (doi:10.1186/s13046-017-0530-4) contains supplementary material, which is available to authorized users.

Regulatory Role of Hexokinase 2 in Modulating Head and Neck Tumorigenesis

“…With extensive studies, including the present investigation, showing that HK2 is a master promoting factor in different stages of carcinogenesis ( Supplementary Table 4), much efforts were put in exploring HK-mediated tumor inhibitors, despite that no potent drugs targeting HK2 are yet available in clinic (80)(81)(82)(83)(84). Taking the most promising HK2-targeting drug, 3-bromopyruvate (3-BP), as an example, although single treatment of 3-BP, at the appropriate dose and formulation, could effectively destroy glycolytic tumors and seems to be non-toxic to all sorts of vertebrates, certain failure cases were still reported in clinical test (83).…”

Benserazide Perturbs Kif15‐kinesin Binding Protein Interaction with Prolonged Metaphase and Defects in Chromosomal Congression: A Study Based on in silico Modeling and Cell Culture

“…The high IC 50 indicated that benserazide is relatively less toxic compared to the anticancer drugs. Several other studies have also reported the toxicity of benserazide in the similar range, for example the IC 50 of benserazide in the cell lines such as SW480, Lovo, HCT116, and MCF7 was 143, 151, 181 and 186 μM respectively . The cytotoxicity of benserazide on HeLa cells shows its potential for repurposing it as an anticancer agent.…”

Ikarugamycin inhibits pancreatic cancer cell glycolysis by targeting hexokinase 2

“…The bioactive compounds that target the Warburg effect to fight cancer can be summarized as being members of two groups: metabolite analogs, for example, 3‐bromopyruvate, and secondary metabolites from natural sources. A series of bioactive natural products have been identified that inhibit HK2, including heterocycles, 2,6‐disubstituted glucosamines, and benserazide . The alkaloids of the PTM class and related compounds are a small group of approximately 30 metabolites with a unique polycyclic skeleton, and macrolactams, which are widely produced by the genera Lysobacter , Saccharophagus, and Streptomyces .…”

“…A series of bioactive natural products have been identified that inhibit HK2, including heterocycles, 2,6-disubstituted glucosamines, and benserazide. 39,40 The alkaloids of the PTM class The ECAR was normalized to the total protein content between groups prior to making comparisons and is reported as mpH/min. F, Quantification of glycolytic capacity.…”