In vitro and in vivo metabolism and pharmacokinetics of a chalcone extract from the hairy cliff bean vine
Natural compounds have made significant contributions to the discovery of new drugs, and flavonoids are a major category of natural products that have attracted much attention due to their excellent anti-inflammatory, anti-tumor, antibacterial, anti ulcer, hepatoprotective, and cardiovascular protective activities. The anti-inflammatory active compound 2,5-dimethoxyfuran [4 “, 5”: 3,4] chalcone, extracted and isolated from the stem of Millettia velutina Dunn in the legume family, belongs to chalcone compounds. The research results of Ma et al. showed that the compound has strong anti-inflammatory activity. In vitro cell experiments, it mainly exhibits good anti-inflammatory activity by inhibiting IL-1 β secretion and Caspase-1 activation, as well as inhibiting ASC aggregation. And it can significantly alleviate LPS induced acute shock in mice in vivo experiments. In the early stages of new drug development, conducting research on drug metabolism can help quickly identify compounds that are safe, effective, and have good pharmacokinetic properties, eliminate unqualified compounds in advance, and save research and development costs. Currently, there have been no reports on in vivo and in vitro metabolic data of this compound in domestic and foreign journals. This study investigated the metabolites of 2,5-dimethoxyfuran [4 “, 5”: 3,4] chalcone in liver microsomes of five species including rats, mice, rhesus monkeys, Beagle dogs, and humans, as well as in rats. The pharmacokinetic studies of 2,5-dimethoxyfuran [4 “, 5”: 3,4] chalcone and its metabolite (M1) in vitro and in vivo were also conducted to explore the main substance basis of the compound, laying the foundation for further mechanism research.

 

 

The hairy cliff bean vine is one of the traditional Chinese medicines with good anti-inflammatory effects. Sample 1, as its main anti-inflammatory component, has received limited research both domestically and internationally, and there have been no reports on its in vitro and in vivo metabolism. The UPLC-QE Orbitrap MS method used in this experiment has high sensitivity and specificity, and has significant advantages in the discovery and identification of metabolites. At the same time, the metabolites of sample 1 in liver microsomes of five species including mice, rats, monkeys, dogs, and humans were identified as M1 and M2, and the metabolites M1 and M2 were found in vivo M2、M3、M4、M5， Afterwards, the structure of each metabolite was inferred based on the mass spectrometry fragments, and M1 was synthesized using synthetic methods, proving the accuracy and reliability of the inferred compound structure. The UFLC-MS/MS method was used for quantitative analysis of in vivo samples. Methodological validation showed that the method is stable and reliable, and can quickly detect trace target compounds in the sample. Compared with liquid chromatography and UV analysis methods, it has higher sensitivity advantages.
The pharmacokinetic parameters of compounds are an important indicator for evaluating compounds. Through in vitro and in vivo studies, the main metabolite is M1. Then, the metabolic rates of sample 1 and M1 were studied in rats, and their pharmacokinetic parameters were calculated. It was found that sample 1 was converted to M1 in vivo and further metabolized to M5. This study investigated the in vivo and in vitro metabolites of sample 1, inferring five metabolites. The pharmacokinetic parameters of sample 1 and the main metabolite M1 were measured in rats, and the changes in the plasma of sample 1 and M1 were explored. The results showed that compound 1 was unstable in vivo and rapidly metabolized into M1, indicating that its metabolites rather than itself played an anti-inflammatory role. This study provides a theoretical basis for elucidating the mechanism of anti-inflammatory substances in vivo.