Xianming Lan

Raphani Semen (RS) has two different application forms in the Chinese Pharmacopoeia (ChP): raw RS and stir-fried RS. However, the compounds of raw and stir-fried RS decoctions, in vivo metabolic pathways, and the effects on promoting gastrointestinal motility remain completely unclear. To address this, a Taxonomy-Driven Core Scaffold Expansion (TDCSE) strategy was developed to track compounds of raw and stir-fried RS decoctions and subsequent in vivo metabolic pathways. Then, a zebrafish model was used to evaluate the effect of raw and stir-fried RS decoctions on promoting gastrointestinal motility. Finally, through network pharmacology and molecular docking techniques, the potential targets and pathways of absorbed compounds were predicted. The results indicated that 26 compounds were detected in the stir-fried RS decoction. In addition to these 26 compounds, another 50 compounds were detected in the raw RS decoction. Moreover, a total of 123 metabolites (including prototypes) were detected after removing the duplications. A total of 18, 71, 59, and 4 metabolites were discovered in plasma, urine, feces, and liver tissue, respectively. Evaluation in a zebrafish model demonstrated that both raw RS and stir-fried RS decoctions possessed the effect of promoting gastrointestinal motility compared with the model group. The promoting gastrointestinal motility potential targets, PTGS2, TNF, PARP1, MTOR, etc., and the IL-17 pathway were predicted. Collectively, this study clarified the differences in compounds and in vivo metabolites of raw and stir-fried RS decoctions. It provided a reference for further investigation into the effect of promoting gastrointestinal motility of RS.

Xanthohumol (XN), a natural prenylated flavonoid extracted and isolated from the hop plant (Humulus lupulus), possesses diverse pharmacological activities. Although the metabolites of XN have been investigated in the previous study, a comprehensive metabolic profile has been insufficient in vivo or in vitro until now. The current study was aimed at systematically elucidating the metabolic pathways of XN after oral administration to rats. Herein, a UHPLC-Q-Exactive Orbitrap MS was adopted for the potential metabolites detection. A stepwise targeted matching strategy for the overall identification of XN metabolites was proposed. A metabolic net (53 metabolites included) on XN in vivo and in vitro, as well as the metabolic profile investigation, were designed, preferably characterizing XN metabolites in rat plasma, urine, liver, liver microsomes, and feces. On the basis of a stepwise targeted matching strategy, the net showed that major in vivo metabolic pathways of XN in rats include glucuronidation, sulfation, methylation, demethylation, hydrogenation, dehydrogenation, hydroxylation, and so on. The proposed metabolic pathways in this research will provide essential data for further pharmaceutical studies of prenylated flavonoids and lay the foundation for further toxicity and safety studies.

Naringenin (5,7,4′-trihydroxyflavanone), belonging to the flavanone subclass, is associated with beneficial effects such as anti-oxidation, anticancer, anti-inflammatory, and anti-diabetic effects. Drug metabolism plays an essential role in drug discovery and clinical safety. However, due to the interference of numerous endogenous substances in metabolic samples, the identification and efficient characterization of drug metabolites are difficult. Here, ultra-high-performance liquid chromatography (UHPLC) coupled with high-resolution mass spectrometry was used to obtain mass spectral information of plasma (processed by three methods), urine, feces, liver tissue, and liver microsome samples. Moreover, a novel analytical strategy named “ion induction and deduction” was proposed to systematically screen and identify naringenin metabolites in vivo and in vitro. The analysis strategy was accomplished by the establishment of multiple “net-hubs” and the induction and deduction of fragmentation behavior. Finally, 78 naringenin metabolites were detected and identified from samples of rat plasma, urine, feces, liver tissue, and liver microsomes, of which 67 were detected in vivo and 13 were detected in vitro. Naringenin primarily underwent glucuronidation, sulfation, oxidation, methylation, ring fission, and conversion into phenolic acid and their composite reactions. The current study provides significant help in extracting target information from complex samples and sets the foundation for other pharmacology and toxicology research.