Studies On Chemical Constituents And Anti-tumor Activity Of Ethyl Acetate Fraction Of Forsythia Suspensa(Thunb.) Vahl

Forsythia suspensa(Thunb.)Vahl is a deciduous shrub of the class Dicotyledoneae,Contortae,Oleaceae,Forsythia.It is a precious Chinese herbal medicine in China.Forsythia suspensa is rich in chemical constituents,mainly phenylethanoid glycosides,lignans,phenolic acids,flavonoids and volatile oils.Forsythia suspensa was originally contained in the’Shennong’s Classic of Materia Medica’.It has the functions of clearing heat and detoxifying,eliminating swelling and dispersing knot and other effects Forsythia sus pensa has been used by medical workers in various fields since ancient times because of its wide range of pharmacological effects,such as anti-inflammatory,anti-bacterial and so on.The research on anti-tumor of Forsythia suspensa also has a good prospect.The combination of traditional Chinese medicine theory and modern high-tech level research will provide more development space for the application of Forsythia suspensa.In this study,75%aqueous ethanol solvent was used to extract Forsythia suspensa by ultrasonic and organic solvent.After that,the chemical constituents of ethyl acetate fraction of Forsythia suspensa were separated and purified by Hp20 column chromatography,silica gel column chromatography,polyamide column chromatography,open ODS column chromatography,Sephadex LH-20 column chromatography,RP-HPLC and so on.The chemical constituents of ethyl acetate fraction of Forsythia suspensa were isolated and purified.The structures of all the compounds were identified by physicochemical properties and spectroscopies.The molecular mechanism of anti-tumor activity of Forsythia suspensa was studied by network pharmacology and molecular docking.The antitumor activity of all compounds were studied by MTT method.The volatile oil components of Forsythia suspensa from six different producing areas were analyzed by GC-MS.Finally,nine isolated compounds were obtained.It contains forsythiayanoside E(1),Epipinoresinol-4-O-β-D-glucopyranoside(2),(+)-pinoresinol-4-O-β-D-glucopyranoside(3),matairesinol-4’-O-β-D-glucopyranoside(4),(+)-epipinoresinol-4’-O-β-D-glucopyranoside(5),forsythoside I(6),kaempferol-3-O-sophoroside(7),kaempferol-3-O-rutinoside(8)and forsythoside A(9).Compound 1 is a new compound which has not been reported in the literatures.The results of network pharmacology and molecular docking confirmed that there are 10 anti-tumor active components such as quercetin,kaempferol and luteolin,which act on 16 key targets such as AKT1,IL6 and CCND1:Participated in 20 signaling pathways such as Pathways in cancer.Molecular docking showed that hydrogen bonding,Pi-cation bonding and hydrophobic interaction might be the mechanisms.The results of anti-tumor activity test showed that,the nine isolated compounds had inhibitory activity on MCF-7 and A-375.The results of GC-MS showed that,the common components of Forsythia suspensa from six different producing areas were α-pinene,β-myrcene,methyl benzoate,limonene,etc Most of the essential oils from Forsythia suspensa are monoterpenoids,aldehydes,etc.In addition,Forsythia suspensa from six different producing areas have both common and characteristic components.The quality of volatile oil components in two kinds of Forsythia suspensa from Heilongjiang and other places was compared for the first time,it is found that the two kinds of Forsythia suspensa in Heilongjiang province contain common volatile oil,and each kind of Forsythia suspensa also has its unique components.In this paper,the chemical constituents,pharmacological effects and GC-MS techniques of Forsythia suspensa were collected.The isolated compounds of Forsythia suspensa were obtained by experimental research methods,the anti-tumor activity of Forsythia suspensa was tested,in order to clarify the anti-tumor material basis of Forsythia suspensa.We expect to provide the material basis and theoretical basis for the discovery of new anti-tumor lead compounds,and the qualitative study of the volatile oil components of Forsythia suspensa expect to provide reference and basis for the quality evaluation of Forsythia suspensa in the future.