The discovery of a non-competitive GOT1 inhibitor, hydralazine hydrochloride, via a coupling reaction-based high-throughput screening assay

Qiqi Wu; Zhongya Sun; Zhifeng Chen; Jingqiu Liu; Hong Ding; Cheng Luo; Mingliang Wang; Daohai Du

doi:10.1016/j.bmcl.2022.128883

The discovery of a non-competitive GOT1 inhibitor, hydralazine hydrochloride, via a coupling reaction-based high-throughput screening assay

Bioorg Med Chem Lett. 2022 Oct 1:73:128883. doi: 10.1016/j.bmcl.2022.128883. Epub 2022 Jul 9.

Authors

Qiqi Wu¹, Zhongya Sun², Zhifeng Chen³, Jingqiu Liu³, Hong Ding³, Cheng Luo¹, Mingliang Wang⁴, Daohai Du⁵

Affiliations

¹ School of Chinese Materia Medica, Nanjing University of Chinese Medicine, 138 Xianlin Road, Qixia, Nanjing 210023, Jiangsu, China; Drug Discovery and Design Center, the Center for Chemical Biology, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.
² Drug Discovery and Design Center, the Center for Chemical Biology, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China; School of Life Science and Technology, Harbin Institute of Technology, Harbin 200092, China.
³ Drug Discovery and Design Center, the Center for Chemical Biology, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.
⁴ Department of Medicinal Chemistry, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China; Zhongshan Institute for Drug Discovery, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Zhongshan 528437, China. Electronic address: wangmingliang@simm.ac.cn.
⁵ School of Chinese Materia Medica, Nanjing University of Chinese Medicine, 138 Xianlin Road, Qixia, Nanjing 210023, Jiangsu, China; Drug Discovery and Design Center, the Center for Chemical Biology, State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China. Electronic address: dhdu@simm.ac.cn.

PMID: 35820623
DOI: 10.1016/j.bmcl.2022.128883

Abstract

Glutamate oxaloacetate transaminase 1 (GOT1) plays a key role in aberrant glutamine metabolism. GOT1 suppression can arrest tumor growth and prevent the development of cancer, indicating GOT1 as a potential anticancer target. Reported GOT1 inhibitors, on the other hand, are quite restricted. Here, we developed and optimized a coupling reaction-based high-throughput screening assay for the discovery of GOT1 inhibitors. By using this screening assay, we found that the cardiovascular drug hydralazine hydrochloride inhibited GOT1 catalytic activity, with an IC₅₀ of 26.62 ± 7.45 μM, in a non-competitive and partial-reversible manner. In addition, we determined the binding affinity of hydralazine hydrochloride to GOT1, with a K_d of 16.54 ± 8.59 μM, using a microscale thermophoresis assay. According to structure-activity relationship analysis, the inhibitory activity of hydralazine hydrochloride is mainly derived from its hydrazine group. Furthermore, it inhibits the proliferation of cancer cells MCF-7 and MDA-MB-468 with a slight inhibitory effect compared to other tested cancer cells, highlighting GOT1 as a promising therapeutic target for the treatment of breast cancer.

Keywords: Glutamate oxaloacetate transaminase 1; High-throughput screening; Hydralazine hydrochloride; Inhibitor.

MeSH terms

Aspartate Aminotransferase, Cytoplasmic* / metabolism
Aspartate Aminotransferase, Cytoplasmic* / pharmacology
Cell Line, Tumor
Cell Proliferation
High-Throughput Screening Assays*
Hydralazine / pharmacology

Substances

Hydralazine
Aspartate Aminotransferase, Cytoplasmic