(-)-Epigallocatechin Gallate is a Noncompetitive Inhibitor of NAD Kinase

Tonghai Liu; Wenjia Shi; Yiluan Ding; Qiqi Wu; Bei Zhang; Naixia Zhang; Mingliang Wang; Daohai Du; Hao Zhang; Bo Han; Dean Guo; Jie Zheng; Qi Li; Cheng Luo

doi:10.1021/acsmedchemlett.2c00163

(-)-Epigallocatechin Gallate is a Noncompetitive Inhibitor of NAD Kinase

ACS Med Chem Lett. 2022 Oct 31;13(11):1699-1706. doi: 10.1021/acsmedchemlett.2c00163. eCollection 2022 Nov 10.

Authors

Tonghai Liu^{1

2

3}, Wenjia Shi⁴, Yiluan Ding², Qiqi Wu⁴, Bei Zhang^{2

3}, Naixia Zhang², Mingliang Wang⁵, Daohai Du², Hao Zhang², Bo Han⁶, Dean Guo², Jie Zheng^{2

3}, Qi Li², Cheng Luo^{1

2

3

4

5}

Affiliations

¹ School of Pharmaceutical Science and Technology, Hangzhou Institute for Advanced Study, UCAS, Hangzhou 310024, China.
² State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.
³ University of Chinese Academy of Sciences, Beijing 100049, China.
⁴ School of Chinese Materia Medica, Nanjing University of Chinese Medicine, Nanjing 210023, China.
⁵ Zhongshan Institute for Drug Discovery, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Zhongshan 528437, China.
⁶ School of Pharmacy/Key Laboratory of Xinjiang Phytomedicine Resource and Utilization, Ministry of Education, Shihezi University, Shihezi 832003, China.

Abstract

Nicotinamide adenine dinucleotide kinase (NADK) controls the intracellular NADPH content and provides reducing power for the synthesis of macromolecules and anti-ROS. Moreover, NADK is considered to be a synthetic lethal gene for KRAS mutations. To discover NADK-targeted probes, a high-throughput screening assay was established and optimized with a Z factor of 0.71. The natural product (-)-epigallocatechin gallate (EGCG) was found to be a noncompetitive inhibitor of NADK with K _i = 3.28 ± 0.32 μΜ. The direct binding of EGCG to NADK was determined by several biophysical methods, including NMR spectroscopy, surface plasmon resonance (SPR) assay, and hydrogen-deuterium exchange mass spectrometry (HDX-MS). The SPR assay showed a K _d of 1.78 ± 1.15 μΜ. The HDX-MS experiment showed that EGCG was bound at the non-substrate-binding sites of NADK. Besides, binding mode prediction and derivative activity analysis revealed a potential structure-activity relationship between EGCG and NADK. Furthermore, EGCG can specifically inhibit the proliferation of KRAS-mutated lung cancer cell lines without affecting KRAS wild-type lung cancer cell lines.