A metabolomic perspective of pazopanib-induced acute hepatotoxicity in mice

Abstract

To elucidate the metabolism of pazopanib, a metabolomics approach was performed based on ultra-performance liquid chromatography coupled with electrospray ionization quadrupole mass spectrometry. A total of 22 pazopanib metabolites were identified in vitro and in vivo. Among these metabolites, 17 were novel, including several cysteine adducts and aldehyde derivatives. By screening using recombinant CYPs, CYP3A4 and CYP1A2 were found to be the main forms involved in the pazopanib hydroxylation. Formation of a cysteine conjugate (M3), an aldehyde derivative (M15) and two N-oxide metabolites (M18 and M20) from pazopanib could induce the oxidative stress that may be responsible in part for pazopanib-induced hepatotoxicity. Morphological observation of the liver suggested that pazopanib (300 mg/kg) could cause liver injury. The aspartate transaminase and alanine aminotransferase in serum significantly increased after pazopanib (150, 300 mg/kg) treatment; this liver injury could be partially reversed by the broad-spectrum CYP inhibitor 1-aminobenzotriazole (ABT). Metabolomics analysis revealed that pazopanib could significantly change the levels of L-carnitine, proline and lysophosphatidylcholine 18:1 in liver. Additionally, drug metabolism-related gene expression analysis revealed that hepatic Cyp2d22 and Abcb1a (P-gp) mRNAs were significantly lowered by pazopanib treatment. In conclusion, this study provides a global view of pazopanib metabolism and clues to its influence on hepatic function.

Identifying the reactive metabolites of tyrosine kinase inhibitors in a comprehensive approach: Implications for drug‐drug interactions and hepatotoxicity

“…Trapping studies performed in vitro evidenced the formation of adducts with methoxylamine and to a lower extent with GSH, suggesting a link between RM formation and CYP TDI . These results concur with the detection of aldehyde intermediates 98a and 98b in vitro and in mice and of cysteine, N ‐acetylcysteine, and glycylcysteine adducts underlying the formation of a p ‐imine‐methide RM in mice (Figure ).…”

“…Otherwise, Choudhury et al 140 highlighted a correlation between GSH depletion and CYP1A2 but not CYP3A4 activity in hepatocyte-like cells. In addition, transaminase elevations were correlated to the amount of 98b, cysteine and glycylcysteine adducts probably arising from a p-imine-methide RM in mice ( Figure 23) and reversed by CYP inhibition, 78 supporting RM involvement in hepatotoxicity. The relationship between RM and hepatotoxicity needs to be confirmed in human.…”

“…and 98b in vitro and in mice 72,78 and of cysteine, N-acetylcysteine, and glycylcysteine adducts underlying the formation of a p-imine-methide RM in mice 78 (Figure 23).…”

“…These results are in agreement with the carboxylic acid metabolite reported by the sponsor in HLM, human hepatocytes and urine, even if its structure is not fully characterized . The formation of 98a and 98b was reported in HLM and in mice urine and feces, with a possible oxidation of 98b into carboxylic acid . The formation of a p ‐imine‐methide upon oxidation of the p ‐methylaniline structural alert, mentioned several times, is supported by the detection of a GSH adduct and of cysteine, N ‐acetylcysteine, and glycylcysteine adducts in mice and human, although this mechanism was nor clearly reported nor discussed (Figure ).…”

Impaired clearance of sunitinib leads to metabolic disorders and hepatotoxicity

“…Ultra-performance LC electrospray ionization quadrupole time-offlight MS (UPLC-ESI-QTOFMS)-based metabolomics was successfully applied to investigate the toxicity mechanisms of TK inhibitors, including pazopanib (Wang et al, 2018), sorafenib (Jensen, Parry, Huang, Beak, et al, 2017), and gefitinib (Liu et al, 2015). Metabolomics has proven to be a powerful tool to investigate drug metabolism and toxicity.…”