Studies On The Components And Activities Of Eupatorium Adenophorum Spreng

Objective:Eupatorium adenophorum Spreng.is a synonym of Ageratina adenophora（Spreng.）,and it is a perennial herbaceous plant belonging to the Asteraceae family（Asteraceae）.As an invasive alien species,it has the high reproductive ability and spreads rapidly and can produce biological toxins or allelochemicals,which has caused great damage to agriculture,animal husbandry and forestry.This subject is to study the chemical constituents of invasive alien species,E.adenophorum Spreng.by using a series of extraction,separation and purification methods to obtain monomer compounds,especially cadinane-type sesquiterpenes.Some of these compounds were screened for their antitumor activities.It is hoped to provide a scientific basis for fully developing E.adenophorum Spreng.,using and consuming these excess resources,and turning it into a treasure.Methods:1.The whole plants of E.adenophorum Spreng.（19 kg）were extracted by 95%Et OH three times at room temperature,the solvent was recovered and the total extract was obtained.The extract was suspended in H₂O and then partitioned successively with petroleum ether,ethyl acetate,and n-butyl alcohol to afford three fractions.Macroporous resin HP 20 column chromatography（CC）,silica gel（CC）,Sephadex LH-20（CC）,TLC and semi-preparative high performance liquid chromatography were used to separate and purify the chemical components.The structures of the monomers were identified by physical and chemical analysis,MS,1H-NMR,¹³C-NMR,2D-NMR,UV,IR,CD,optical rotations and other methods.2.In this study,hepatocarcinoma cells Hep G-2 and Bel-7402,laryngeal cancer cells Hep-2,and esophageal cancer cells Eca-109 were used to conduct in vitro cytotoxicity experiments on some monomeric compounds using MTT method to study the antitumor activity of the compounds.The glucose uptake experiment of L6 skeletal muscle cells was carried out on some compounds,so as to screen the antidiabetic activity of these compounds.Results:1.35 compounds were isolated from E.adenophorum Spreng.Their structures are determined as 9-oxo-10,11-dehydroageraphorone（1）,muurol-4-en-3,8-dione（2）,9-hydroxy-ageraphorone（3）,amorph-4-en-3,8-dione（4）,（+）-（5R,7S,9R,10S）-2-oxocadinan-3,6（11）-dien-12,7-olide（5）,（-）-（5R,6R,7S,9R,10S）-cadinan-3-ene-6,7-diol（6）,1,6-dihydroxy-1-isopropyl-4,7-dimethyl-3,4-dihydronaphthalen-2（1H）-one（7）,（1S,4R）-7-hydroxycala-menen-3-one（8）,（1S,2R,4R）-1-isopropyl-4,7-dimethyl-1,2,3,4-tetrahydrona-phthalene-2,6-diol（9）,（1R,4S）-4,7-dihydroxy-4-isopropyl-1,6-dimethyl-1,2-dihydronaphthalen-2（1H）-one（named:eupatorinol,10）,（1R,6R,10S）-7-（2-hydroxypropan-2-yl）-4,10-dimethyl-1,2,3,6-tetrahydronaphthalen-2（1H）-one（named:eupatorinone A,11）,（1R,3R,6S,7R,10S）-3-hydroxy-7-isopropyl-4,10-dimethyl-1,2,3,6,9,10-hexahydronaphthalen-2（1H）-one（named:eupator-inone B,12）,（1S,4S,10R）-7-isopropyl-4,10-dimethyl-1,2,4,5,9,10-hexahydro-naphthalene-3,8-dione（named:eupatorinone C,13）,（1S,3S,6R,10R）-3-hydro-xy-4,10-dimethyl-7-（propan-2-ylidene）-1,2,3,6,9,10-hexahydronaphthalen-2（1H）-one（named:eupatorinone D,14）,（-）-isochaminic acid（15）,thymoquinol5-O-β-D-glucopyranoside（16）,thymoquinol 2-O-β-D-glucopyranoside（17）,eupatonriochromene（18）,1-（8-hydroxy-2,2-dimethyl-2H-chromen-6-yl）ethan-1-one（19）,7-hydroxyl-4-methyl isocoumarine（20）,demethoxyencecalin（21）,encecalin（22）,7-hydroxy-dehydrotremetone（23）,2R*,3S*-toxol-7-O-β-D-glucopyranoside（24）,3β-hydroxy-urs-11-en-28,13β-olide（25）,ψ-taraxasterone（26）,stigmasterol（27）,patuletin-4’-methoxyl-7-O-β-D-glucopyranoside（28）,6-hydroxykaempferol-7-O-β-D-glucopyranoside（29）,7-O-（6-methoxykaempferol）-O-β-D-glucopranoside（30）,trans-melilo-toside（31）,cis-melilotoside（32）,protocatechuic acid（33）,p-Hydroxybenzoic acid（34）,lignoceric acid（35）.Among them,compounds 1-14 were cadinane-type sesquiterpenes.Compounds 10-14 are new compounds,and their absolute configuration is determined by NMR calculation and ECD.2.The results of in vitro anti-tumor activity screening showed that the IC₅₀ values of esophageal cancer cells Eca-109 were 111.000±10.635,58.542±2.394,41.392±1.928μg/m L and 118.040±5.523,61.746±1.317,35.628±0.710μg/m L,respectively,after treatment with compounds 22 and25 at 12,24 and 48 h.The results of screening the glucose uptake activity of L6 cells were that compounds 2,11 and 13 increased the glucose uptake of L6cells by 1.600,1.426 and 1.214 times,respectively.Conclusion:Compounds 10-14 isolated from E.adenophorum,which are new compounds.Compounds 8,15,20,26,28 were isolated from genus Eupatorium for the first time,and compounds 16,17,18,24,25,29 were isolated from E.adenophorum for the first time.The results of the activity study showed that the compounds 22 and 25 have anti-tumor activity against esophageal cancer cells Eca-109.The screening results of glucose uptake activity of L6 skeletal muscle cells show that compounds 2,11,and 13 can promote glucose uptake of L6 cells in vitro.