Identification of osalmid metabolic profile and active metabolites with anti-tumor activity in human hepatocellular carcinoma cells

作者：Zhe Wu, Yaqiong Zhan, Li Wang, Jiepeng Tong, Li Zhang, Mengjia Lin, Xuehang Jin, Lushun Jiang, Yan Lou, Yunqing Qiu · 发表于：Biomedicine & Pharmacotherapy · 年份：2020 · DOI：10.1016/j.biopha.2020.110556 · 被引用次数：16 · 研究领域：Metal-Catalyzed Oxygenation Mechanisms、RNA modifications and cancer、Cancer, Hypoxia, and Metabolism

BACKGROUNDS: Ribonucleotide reductase (RR) catalyzes the essential step in the formation of all four deoxynucleotides. Upregulated activity of RR plays an active role in tumor progression. As the regulatory subunit of RR, ribonucleotide reductase subunit M2 (RRM2) is regarded as one of the effective therapeutic targets for DNA replication-dependent diseases, such as cancers. Recent studies have revealed that osalmid significantly inhibits the activity of RRM2, but the metabolic profile of osalmid remains unknown. OBJECTIVE: The aim of this study was to clarify the metabolic profile including metabolites, isoenzymes and metabolic pathways of osalmid. The anti-human hepatocellular carcinoma activity and mechanism of metabolites were further investigated. MATERIALS AND METHODS: Ultra high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC/Q-TOF-MS) was used for identifying metabolites and for characterizing phase I and phase II metabolic pathways with recombinant enzymes or in human liver microsomes of osalmid. The eHiTS docking system was used for potential RRM2 inhibitor screening among metabolites. Cytotoxicity assays were performed for evaluating cell proliferation inhibitory activity of metabolites. Cell cycle assays and cell apoptosis assays were assessed by flow cytometry. Western blotting analysis of RRM2, cyclin D1, p21, p53, phosphorylated p53, Bcl-2 and Bax was performed to explore the anti-hepatocellular carcinoma mechanism of the act...