Chemical Constituents From The EtOAc Fraction Of Cortex Moutan And Their Anti-tumor Activities

Objective:Study the chemical constituents of Cortex Moutan（Paeonia Suffruticosa Andrews）,conduct pharmacodynamics,network pharmacology and molecular docking experiments,predict possible targets and pathways,screen natural compounds with potential anti-tumor activity.The research provides basis for the study of anti-tumor pharmacodynamics and rational development of Cortex Moutan.Methods:1.The dried medicinal materials of Cortex Moutan were pulverized,75%Et OH was used for backflow fraction,and the fraction was distilled under decompress to get the Et OH fraction.The solvent gradient extraction method was adopted,followed by extraction with PE,CH₂Cl₂,Et OAc and n-Bu OH,and four fractions were obtained by vacuum distillation.All kinds of chromatographic separation technology of chemical components were applied to detach the chemical constituents from the Et OAc fraction of Cortex Moutan with good activity,and the structural identification of compounds by nuclear magnetic resonance spectroscopy(¹H-NMR and ¹³C-NMR)and other techniques.2.The anti-tumor activity of Cortex Moutan was evaluated by MTT assay.The cell viability rates of the human breast cancer MDA-MB-231 cell line,the human lung adenocarcinoma A549 cell line and the human liver cancer SMMC-7721 cell line were compared with the positive control drug Taxol by applying the Cortex Moutan extract and the isolated monomer compounds,compared and judged its anti-tumor activity.3.The network pharmacology method and molecular docking technology were adopted to study the target and action mechanism of Cortex Moutan in the treatment of tumor diseases.Results:1.In the process of researching the chemical component of the Et OAc fraction of Cortex Moutan,a total of 24 compounds were isolated,and the structures were determined by ¹H-NMR and ¹³C-NMR,their structures were identified as benzoic acid（1）,4-hydroxybenzoic acid（2）,paeonol（3）,vanillic acid（4）,ethyl3,5-dihydroxy-4-methoxybenzoate（5）,gallic acid（6）,methyl gallate（7）,ethyl gallate（8）,ferulic acid（9）,isoferulic acid（10）,ethyl 3-（4-hydroxy-3-methoxyphenyl）acrylate（11）,paeoniflorin（12）,benzoylpaeoniflorin（13）,hederagenin（14）,methyl hederagenin（15）,kalopanaxsaponin A（16）,oleanolic acid（17）,betulinic acid（18）,naringenin（19）,quercetin（20）,catechin（21）,stigmasterol（22）,daucosterol（23）,6-hydroxy-7-methoxycoumarin（24）.There were 11 phenolic compounds（1-11）;7terpenoids（12-18）;3 flavonoids（19-21）;2 sterols（22-23）;1 coumarin compound（24）.Compounds 5,11,15,16,22,and 24 were isolated from Cortex Moutan for the first time.2.In vitro test results showed that the Et OAc fraction from Cortex Moutan had better anti-tumor activity.Compounds 3,4,6,16,17,20,22 and 24 had anti-tumor activity in MDA-MB-231 cells;compounds 3,6,16,18 and 20 had certain anti-tumor activity against A549 cells;the inhibitory effect of compound 3,6,15,16,17,18,19and 20 on SMMC-7721 cells inhibited the proliferation of tumor cells.Compounds 3,6,16 and 20 exhibited better anti-tumor activity than positive control drug Taxol in the viability rate of three cell lines（MDA-MB-231,A549 and SMMC-7721）,among which compound 16 had the strongest anti-tumor activity.3.The effects for network pharmacology displayed that flavonoids and monoterpenoids of Cortex Moutan were associated with many anti-tumor targets,EGFR and CTNNB1 were the key targets in the protein interaction network.The anti-tumor pathways include PI3K-Akt and Ras signaling pathways.The molecular docking results showed that the binding index of quercetin to EGFR was the highest,followed by benzoylpaeoniflorin,suggesting that quercetin and benzoylpaeoniflorin might be potential inhibitors of EGFR.Conclusion:In this study,24 compounds were isolated from the Et OAc fraction of Cortex Moutan,which provided a basis for further research on the pharmacological mechanism of Cortex Moutan and its rational development and utilization.In vitro test results proved that Cortex Moutan has anti-tumor effect.Network pharmacology and molecular docking were used to further explore the anti-tumor mechanism of Cortex Moutan,which preliminarily revealed that Cortex Moutan may synergistically exert anti-tumor effects through multiple components,targets and signaling pathways,laying a foundation for its in-depth research,and also for the development and application of Cortex Moutan provides a theoretical basis.