The Metabolism of Separase Inhibitor Sepin-1 in Human, Mouse, and Rat Liver Microsomes

Abstract

Separase, a known oncogene, is widely overexpressed in numerous human tumors of breast, bone, brain, blood, and prostate. Separase is an emerging target for cancer therapy, and separase enzymatic inhibitors such as sepin-1 are currently being developed to treat separase-overexpressed tumors. Drug metabolism plays a critical role in the efficacy and safety of drug development, as well as possible drug–drug interactions. In this study, we investigated the in vitro metabolism of sepin-1 in human, mouse, and rat liver microsomes (RLM) using metabolomic approaches. In human liver microsomes (HLM), we identified seven metabolites including one cysteine–sepin-1 adduct and one glutathione–sepin-1 adduct. All the sepin-1 metabolites in HLM were also found in both mouse and RLM. Using recombinant CYP450 isoenzymes, we demonstrated that multiple enzymes contributed to the metabolism of sepin-1, including CYP2D6 and CYP3A4 as the major metabolizing enzymes. Inhibitory effects of sepin-1 on seven major CYP450s were also evaluated using the corresponding substrates recommended by the US Food and Drug Administration. Our studies indicated that sepin-1 moderately inhibits CYP1A2, CYP2C19, and CYP3A4 with IC50 < 10 μM but weakly inhibits CYP2B6, CYP2C8/9, and CYP2D6 with IC50 > 10 μM. This information can be used to optimize the structures of sepin-1 for more suitable pharmacological properties and to predict the possible sepin-1 interactions with other chemotherapeutic drugs.

Metabolism of a Selective Serotonin and Norepinephrine Reuptake Inhibitor Duloxetine in Liver Microsomes and Mice

“…In this study, we investigated the metabolic pathways of DLX in human liver microsomes (HLM) /mouse liver microsomes (MLM) and mice using liquid chromatography mass spectrometry (LC-MS)-based metabolomic approaches. Our previous studies demonstrated that LC-MS-based metabolomics is a rapid and effective approach to investigate drug metabolism ( Li et al, 2018 ; Li et al, 2020 ), bioactivation ( Li et al, 2011b ; Li et al, 2014 ; Liu et al, 2015 ; Liu et al, 2016 ; MacKenzie et al, 2020 ), and toxicity (O'Connell and Watkins, 2010; Li et al, 2013 ; Lu et al, 2019 ; Zhao et al, 2019 ). Here, a total of 39 metabolites generated from DLX were identified, of which 13 metabolites are novel.…”

The roles of Cyp1a2 and Cyp2d in pharmacokinetic profiles of serotonin and norepinephrine reuptake inhibitor duloxetine and its metabolites in mice

“…2A showed that Cyp1a2 protein expression is absent in Cyp1a2-KO mouse livers. The O-deethylation of phenacetin is regarded as one of the probe reactions primarily catalyzed by human CYP1A2 or mouse Cyp1a2 ( Li et al, 2018 ). The formation of O-deethylation-phenacetin was suppressed by 80% in LS9 from Cyp1a2-KO mice compared with WT mice ( Fig.…”

“…To test if propranolol has the inhibitory effects on Cyp2d in our mouse model used in the present study, 16 h after propranolol treatment the LS9s were prepared from mice. The Cyp2d function was first evaluated using Cyp2d probe dextromethorphan ( Li et al, 2018 ). No significant difference was observed between WT + control and KO + control groups, indicating that Cyp1a2-KO had no effect on Cyp2d activity ( Fig.…”

“…The selective probe phenacetin was used to validate the function of Cyp1a2 in the liver from WT and Cyp1a2-KO mice ( Li et al, 2018 ). Liver S9 fraction (LS9) was prepared by adding 10 folds of pre-chilled 1x PBS (w/v) to the livers from male WT or Cyp1a2-KO mice aged between 8 and 12 weeks (n = 4).…”

Metabolic profiling of norepinephrine reuptake inhibitor atomoxetine

“…Metabolomics has proven to be an effective and powerful tool to investigate drug metabolism (Li et al, 2018;Li et al, 2020), bioactivation (Li et al, 2014;Liu et al, 2015;Liu et al, 2016), and drug toxicity (Li et al, 2013;Lu et al, 2019;Zhao et al, 2019). We identified 31 metabolites and adducts generated from ATX in mice and LMs, of which 16 are novel.…”