A comparative study on the in vitro biotransformation of medicagenic acid using human liver microsomes and S9 fractions. (1st September 2020)

• Record Type: Journal Article
• Authors: Peeters, Laura; Vervliet, Philippe; Foubert, Kenn; Hermans, Nina; Pieters, Luc; Covaci, Adrian
• Is Part Of: Chemico-biological interactions. Volume 328(2020)
• Publication Date: 2020-09-01
• Subjects: In vitro liver biotransformation -- Medicagenic acid -- Non-target screening -- Human liver microsomes -- Human liver S9; Biochemistry -- Periodicals; Toxicological chemistry -- Periodicals; Biochemistry -- Periodicals; Biologie moléculaire -- Périodiques; Biochimie -- Périodiques; Toxicologie biochimique -- Périodiques; 572
• DOI: 10.1016/j.cbi.2020.109192
• Languages: English
• ISSNs: 0009-2797

Abstract

Abstract: Many natural products are prodrugs which are biotransformed and activated after oral administration. The investigation of gastrointestinal and hepatic biotransformation can be facilitated by in vitro screening methods. This study compares two widely used in vitro models for hepatic biotransformation: 1) human S9 fractions and 2) human liver microsomes and cytosolic fractions in a two-step sequence, with the purpose of identifying differences in the biotransformation of medicagenic acid, the putative precursor of active metabolites, responsible for the medicinal effects of the herb Herniaria hirsuta . The combination of liquid chromatography coupled to high-resolution mass spectrometry with subsequent suspect and non-target data analysis allowed the identification of thirteen biotransformation products, four of which are reported here for the first time. Eight biotransformation products resulting from oxidative Phase I reactions were identified. Phase II conjugation reactions resulted in the formation of three glucuronidated and two sulfated biotransformation products. No major differences could be observed between incubations with human liver S9 or when utilizing human microsomal and cytosolic fractions. Apart from two metabolites, both methods rendered the same qualitative metabolic profile, with minor quantitative differences. As a result, both protocols applied in this study can be used to study in vitro human liver biotransformation reactions. Graphical abstract: Image 1 Highlights: Comparison of S9 and microsomal + cytosolic fractions for in vitro liver biotransformation. Aim: Identifying differences in biotransformation of medicagenic acid. 13 biotransformation products were identified, 4 are reported for the first time. Similar qualitative metabolic profile with minor quantitative differences. Both protocols can be used to study in vitro human liver biotransformation.