Characterization of in vitro metabolites of irisflorentin by rat liver microsomes using high‐performance liquid chromatography coupled with tandem mass spectrometry

Abstract

Belamcanda chinensis has been extensively used as antibechic, expectorant and anti-inflammatory agent in traditional medicine. Irisflorentin is one of the major active ingredients. However, little is known about the metabolism of irisflorentin so far. In this work, rat liver microsomes (RLMs) were used to investigate the metabolism of this compound for the first time. Seven metabolites were detected. Five of them were identified as 6,7-dihydroxy-5,3',4',5'-tetramethoxy isoflavone (M1), irigenin (M2), 5,7,4'-trihydroxy-6,3',5'-trimethoxy isoflavone (M3), 6,7,4'-trihydroxy-5,3',5'-trimethoxy isoflavone (M4) and 6,7,5'-trihydroxy-5,3',4'-trimethoxy isoflavone (M5) by means of NMR and/or HPLC-ESI-MS. The structures of M6 and M7 were not elucidated because they produced no MS signals. The predominant metabolite M1 was noted to be a new compound. Interestingly, it was found to possess anticancer activity much higher than the parent compound. The enzymatic kinetic parameters of M1 revealed a sigmoidal profile, with Vmax = 12.02 μm/mg protein/min, Km = 37.24 μm, CLint = 0.32 μL/mg protein/min and h = 1.48, indicating the positive cooperation. For the first time in this work, a new metabolite of irisflorentin was found to demonstrate a much higher biological activity than its parent compound, suggesting a new avenue for the development of drugs from B. chinensis, which was also applicable for other herbal plants. Copyright © 2016 John Wiley & Sons, Ltd.

<p>Metabolite Profiling in Anticancer Drug Development: A Systematic Review</p>

“…Out of 78 articles included in the analysis (42 in vitro, 42 in vivo, and 16 clinical studies), 47 (60.3%), 14 (17.9%) and 17 (21.8%) articles respectively, investigated metabolite profiles of conventional anticancer drugs/synthetic anticancer candidates, small-molecules targeted therapy, and herbderived compounds with anticancer activities. These studies involved a total of 57 (57.0%) conventional anticancer drugs/ synthetic anticancer candidates, 6, 22 (22.0%) studies for small-molecules targeted therapy, 4,5,[59][60][61][62][63][64][65][66][67][68][69][70] and 21 (21.0%) studies for herb-derived compounds with anticancer activities 2,3,7,9,[71][72][73][74][75][76][77][78][79][80][81][82][83] (Tables 1-3). Anticancer drugs or candidate compounds are metabolized by either Phase I, or phase II metabolizing enzyme alone, or both phase I and phase II metabolizing enzymes.…”

“…Relationship between the generated metabolites and anticancer activity and/or toxicity were reported in 10 (17.5%) studies for conventional anticancer drugs/synthetic anticancer candidates, 6,13,22,31,32,36,45,50 1 (4.5%) studies for smallmolecules targeted therapy, 65 and 4 (19.0%) studies for herb-derived compounds with anticancer activities. 75,76,78,79…”

“…75 The M1 and M2 metabolites of the synthetic compound irisflorentin were shown to exhibit more potent cytotoxic activity against human prostate cancer (DU145) and breast cancer (MCF-7) cell lines compared with the parent drug (IC 50 65.12 vs. 35.71 vs.>300 µM and 74.17 vs. 36.30 vs. >300 µM, respectively). 78 The metabolite of a potent antileukemic compound, phyllanthoside, also exhibited lower cytotoxic activity against human rhabdomyosarcoma A204 cell line compared with the parent compound (IC 50 of 24 µM and 0.47 nM, respectively). 50 Some metabolites showed low or no cytotoxic activity.…”

Application of ultra‐performance liquid chromatography coupled with quadrupole time‐of‐flight mass spectrometry in identification of three isoflavone glycosides and their corresponding metabolites

“…Isoflavones, the secondary metabolites of herbal medicine, possess a variety of bioactivities such as antitumor, anti‐inflammatory, antioxidative, antiviral, antibacterial, and estrogenic effects . Recently, metabolites of isoflavones in the human body have aroused great interest due to the potential higher bioactivities, which became one of the most efficient ways for drug discovery . The intestinal flora in the gastrointestinal tract exert an important influence on the in vivo biotransformation of the components in phytomedicines .…”

Biotransformações dos ácidos ent-poliáltico e ent-diidroagático por culturas microbianas e microssomas hepáticos

“…O desenvolvimento de modelos in vitro mais recentes levam em consideração a arquitetura dos órgãos in vivo e a integração do trato gastointestinal com o fígado, a fim de se obter sistemas mais próximos possíveis das condições reais. microssomas são modelos de fácil utilização e também podem ser úteis na pesquisa do metabolismo de produtos naturais bioativos(ZENGER et al, 2015;BENDIKOV et al, 2016, JIA et al, 2016. Sua utilização também permite a análise de variações metabólicas interindividuais e populacionais, apesar da extrapolação quantitativa da formação de metabólitos nesses modelos serem consideradas limitadas(FASINU, BOUIC & ROSENKRANZ, 2012).…”