Characterization of metabolism feature and potential pharmacological changes of morusin-a promising anti-tumor drug-by ultra-high-performance liquid chromatography coupled time-of-flight mass spectrometry and network pharmacology

Abstract Morusin, a prenylated flavone isolated from Morus species, was treated as a potential anti-tumor drug since it inhibited effects on numerous types of human cancer cells. In some sense, drugs-related metabolites always contribute to pharmacological changes, inducing therapy improvement, reduced efficacy, or side-effects. Thus, the characterization of metabolites and the potential functions exerted great importance in clinical applications. Till now, the metabolism feature of morusin was still unclear, and its pharmacological changes were still unconducted, too. In this work, an integrated strategy based on metabolites profiling and network pharmacological was applied to characterize the metabolism feature and reveal pharmacological changes of morusin in vivo. As a result, a total of 31 metabolites (19 in plasma, 8 in urine, 30 in feces, 6 in heart, 17 in liver, 4 in spleen, 6 in lung, 6 in kidney, and 3 in brain) were screened out in rats, and 11 of them were characterized for the first time. Among them, metabolites M6, M18, M19, M20, and M28, were the main metabolites. Phase I metabolic reactions of hydroxylation, dehydrogenation (cyclization), isomerization and phase II reactions of glucuronidation occurred, and glucuronidation and hydroxylation were the two main metabolic reactions. Moreover, the pharmacological difference between morusin and five main metabolites was predicted by an network pharmacological method, and these 6 candidates targeted 177 targets. Meanwhile, in addition to common pathways (PI3K-Akt signaling pathway, proteoglycans in cancer, hepatitis B, cAMP signaling pathway, and viral carcinogenesis) of morusin in cancer, six metabolites’ targets were involved in prostate cancer, chemokine signaling pathway, ras signaling pathway and neuroactive ligand-receptor interaction, indicating that these functional changes might result in novel pharmacological mechanism and new indications. Our work provided the metabolism feature and functional modifications of morusin in vivo for the first time, and meaningful information for further pharmacological validations or potential indications in clinic were supplied.