Pharmacokinetic and Relative Bioavailability of Three Secoiridoid Glycosides in Beagle Dog Plasma After Oral Administration of Conventional and Enteric-Coated Capsules of Gentianella acuta Extract

Objective:To simultaneously investigate the pharmacokinetics of gentiopicroside,sweroside,and swertiamarin,which are constituents of Gentianella acuta,by developing and validating a simple,sensitive,and fast ultra-high-performance liquid chromatography–tandem mass spectrometry method.Materials and Methods:Blood samples were collected from the forward limb veins of six beagle dogs following oral gavage with G.acuta,the whole plant extract(39.90 mg/kg).Plasma samples were processed using liquid–liquid extraction.The analytes and paeoniflorin(internal standard[IS])were separated using an Acquity?UPLC ethylene bridged hybrid amide column(2.1 mm×100 mm,1.7μm)with isocratic elution using a mobile phase consisting of acetonitrile and 0.1%formic acid in water(80:20,v/v)at a flow rate of 0.4 mL/min.Quantification was performed using multiple reaction monitoring of the fragmentation transitions at m/z 401.1→179.0,403.1→195.0,419.1→179.0,and 525.2→449.1 for gentiopicroside,sweroside,swertiamarin,and the IS,respectively.Results:The linearity of the analytical response was good and the calibration curves were linear over concentration ranges of 1.20–192.0,0.40–159.0,and 0.20–209.3 ng/mL for gentiopicroside,sweroside,and swertiamarin,respectively.The extraction recovery was in the range of 84.72%–91.34%,84.58%–93.43%,and 82.75%–91.37%for gentiopicroside,sweroside,and swertiamarin,respectively.Conclusions:The method was successfully used to evaluate the pharmacokinetic parameters of gentiopicroside,sweroside,and swertiamarin in beagle dogs.

Simultaneous Determination and Pharmacokinetics of Tetrandrine,Fangchinoline,and Cyclanoline in Rat Plasma by Ultra-High Performance Liquid Chromatography-Mass Spectrometry after Oral Administration of Stephaniae Tetrandrae Radix Extract

Objective:The objective of the study was to develop a rapid and sensitive ultra-performance liquid chromatography-tandem mass spectrometric method for the determination of tetrandrine,fangchinoline,and cyclanoline in rat plasma and to investigate their pharmacokinetics after oral administration of Stephaniae Tetrandrae Radix extracts.Methods:Sample pretreatment involved methanol pretreatment and liquid–liquid extraction of ethyl acetate from plasma with methanol.Tramadol was used as the internal standard.The analysis was performed using an high strength silica T3 column(100 mm×2.1 mm,1.8μm)and a gradient elution method consisting of mobile phase solution A(0.1%formic acid in water)and B(acetonitrile)at a flow rate of 0.4 mL/min.The detection was performed using a triple quadrupole tandem mass spectrometer in the multiple reaction monitoring mode and using an electrospray ionization source in the positive ionization mode.Results:High efficiency was achieved with an analysis time of 4 min/sample.The calibration curve linear in the concentration range of 1250 ng/ml(R^(2)≥0.9900)and the lower limit of quantification is 1 ng/ml.The intraday and interday precision(relative standard deviation)values were lower than 9.4.Accuracy(relative error)was within 10.3%at all three quality control levels.Conclusions:This method was successfully applied in pharmacokinetics of tetrandrine,fangchinoline,and cyclanoline in rats after oral administration of Stephaniae Tetrandrae Radix extracts.The maximum plasma concentration(C_(max))of tetrandrine,fangchinoline,and cyclanoline was 124.71±16.08,84.56±3.28,and 57.61±6.26 ng/mL,respectively.The time to reach C_(max)was 10.39±3.04 for tetrandrine,10.17±3.04 for fangchinoline,and 6.40±3.16 for cyclanoline.The pharmacokinetic results might help further guide the clinical application of Stephaniae Tetrandrae Radix.

Exploring the Molecular Mechanism of Radix Astragali on Colon Cancer Based on Integrated Pharmacology and Molecular Docking Technique

Objective:The objective of this study was to study the mechanism of Radix Astragali on colon cancer by integrated pharmacology and molecular docking technique.Methods:Integrative pharmacology-based research platform of traditional Chinese medicine(TCMIP)V2.0 was used to obtain the chemical components and corresponding targets of Radix Astragali and the target information of colon cancer to create the main target network of drugs and diseases.Gene Ontology(GO),Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis was carried out using Hiplot website,and the interaction network of“Traditional Chinese Medicine-component-target-pathway”was established,and molecular docking with main targets was carried out for the key components.Results:Twenty-seven chemical constituents of Radix Astragali,their 254 corresponding targets,and 44 colon cancer-related targets were obtained.Through proteins interacting,70 nodes were obtained as core targets.GO analysis showed that it mainly acts on lipid metabolism,nuclear receptor activity,phagocytic cup,etc.KEGG pathway analysis showed that it was mainly enriched in the estrogen signaling pathway,C-type lectin receptor signaling pathway,PI3K-Akt signaling pathway,etc.The multidimensional network,quantitative estimate of the drug,and molecular docking showed that the main targets are AKT1,BCL2,and CDK6,and the key components involved are kumatakenin,astragaloside VIII,and choline.Conclusion:Kumatakenin,AstragalosideⅧ,Choline and other compounds of Radix Astragali may affect colon cancer by acting on AKT1,BCL2 and other targets,thereby regulating estrogen signaling pathway,C-type lectin receptor signaling pathway,PI3K-Akt signaling pathway and so on.Those will provide theoretical reference for future research on the material basis and mechanism of its pharmacodynamics.