Discovery and characterization of a novel glucose-6-phosphate dehydrogenase (G6PD) inhibitor via high-throughput screening

Zhongyuan Luo; Daohai Du; Yanjun Liu; Tian Lu; Liping Liu; Hualiang Jiang; Kaixian Chen; Changliang Shan; Cheng Luo

doi:10.1016/j.bmcl.2021.127905

Discovery and characterization of a novel glucose-6-phosphate dehydrogenase (G6PD) inhibitor via high-throughput screening

Bioorg Med Chem Lett. 2021 May 15:40:127905. doi: 10.1016/j.bmcl.2021.127905. Epub 2021 Mar 6.

Authors

Zhongyuan Luo¹, Daohai Du¹, Yanjun Liu², Tian Lu², Liping Liu², Hualiang Jiang², Kaixian Chen², Changliang Shan³, Cheng Luo⁴

Affiliations

¹ School of Chinese Materia Medica, Nanjing University of Chinese Medicine, 138 Xianlin Road, Qixia, Nanjing 210023, Jiangsu, China; The Center for Chemical Biology, Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zuchongzhi Road, Shanghai 201203, China.
² The Center for Chemical Biology, Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zuchongzhi Road, Shanghai 201203, China.
³ State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy and Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin 300350, China. Electronic address: changliangshan@nankai.edu.cn.
⁴ School of Chinese Materia Medica, Nanjing University of Chinese Medicine, 138 Xianlin Road, Qixia, Nanjing 210023, Jiangsu, China; The Center for Chemical Biology, Drug Discovery and Design Center, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zuchongzhi Road, Shanghai 201203, China; School of Pharmaceutical Science and Technology, Hangzhou Institute for Advanced Study, UCAS, Hangzhou 310024, China. Electronic address: cluo@simm.ac.cn.

PMID: 33689874
DOI: 10.1016/j.bmcl.2021.127905

Abstract

Altered glucose-6-phosphate dehydrogenase (G6PD) status is influential in many cellular pathophysiological processes and diseases, making G6PD a potential target for cancer therapy. However, the available G6PD inhibitors are very limited and restricted. Here we developed a reducing equivalent nicotinamide adenine dinucleotide phosphate (NADPH) absorption photometry assay based on enzyme kinetics to characterize G6PD activity. In this way, we performed a high-throughput screening (HTS) to an in house library. And then we identified compound named Wedelolactone inhibiting G6PD strongly in a non-competitive, reversible way. In addition, we did the surface Plasmon Resonance (SPR) assay and indicated the K_D between Wedelolactone and G6PD protein was 3.64 μM. Furthermore, our basic colony formation assay showed the inhibitory effect of Wedelolactone on the proliferation of ovarian cancer cells (IC₅₀ ~ 10 µM). Thus, we provided a high-throughput screening assay to quickly and efficiently discover G6PD inhibitors, and identified Wedelolactone as a G6PD inhibitor, implying that Wedelolactone suppresses ovarian cancer partly through targeting G6PD.

Keywords: Absorption photometry assay; Enzyme kinetics assay; Glucose-6-phosphate dehydrogenase inhibitor; High-throughput screening.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Antineoplastic Agents / chemistry*
Antineoplastic Agents / pharmacology
Apoptosis / drug effects
Cell Line, Tumor
Cell Proliferation / drug effects
Coumarins / chemistry*
Coumarins / pharmacology
Drug Screening Assays, Antitumor
Enzyme Inhibitors / chemistry*
Enzyme Inhibitors / pharmacology
Female
Glucosephosphate Dehydrogenase / antagonists & inhibitors*
High-Throughput Screening Assays
Humans
NADP / metabolism
Ovarian Neoplasms / drug therapy*
Oxidation-Reduction
Protein Binding
Structure-Activity Relationship
Surface Plasmon Resonance

Substances

Antineoplastic Agents
Coumarins
Enzyme Inhibitors
wedelolactone
NADP
Glucosephosphate Dehydrogenase