| The genus Achillea(Compositae)is composed of 200 species of perennial herbs,all native to temperate regions of the northern hemisphere.Many Achillea species are used in folk medicine as anti-inflammatory,diaphoretic,diuretic,anti-spasmodic and for treatment of menalgia,rheumatic pain and hemorrhage.Phytochemical investigations of Achillea species have revealed many components such as sesquiterpenoids,flavonoids,lignans and essential oil.Modern pharmacological research demonstrated that the extracts of most of Achillea species displayed anti-inflammatory,antibacterial,antitumor and antispasmodic activities.Achillea alpina L.is a perennial herb which belongs to the tribe of Achillea.It is mainly distributed in Inner Mongolia,Hebei,Shanxi,Ningxia,the east of Gansu provinces and northeast of China,also exists in Siberia,Japan,Korea,Mongolia and the Far East.Accoding to the Chinese Pharmacopeia(2015 edition),Achillea alpina L.has the effects of eliminating dampness,detoxication,promoting blood circulation and relieving pain.It is usually used for the treatments of tonsillitis,pharyngalgia,diarrhea,intestinal carbuncle,heat stranguria,damp-heat,insect and snake bite.As a traditional Chinese herbal medicine recorded in Chinese Pharmacopeia,there are only a few reports about the constituents of this plant.In order to promote the exploitation and utilization of Achillea species,we studied the aerial part of Achillea alpine L.and systematically separated the petroleum ether,dichloromethane,ethyl acetate and n-butanol extractions,respectively.And we identified the structures of the compounds and screened their pharmacological activities subsequently.Objective: The aim is to study chemical constituents from the aerial parts of Achillea alpina L.by using various techniques such as column chromatography,preparative thin layer chromatography,preparative high performance liquid chromatography and recrystallization purification.Theirstructures were identified on basis of chemical methods and spectral data such as UV,MS,1D NMR,2D NMR.And then,screening some new,active agents by MTT colorimetric method were carried out.Methods: The air-dried aerial parts of Achillea alpina L.(9.3 kg)were crushed and extracted with 95% ethanol by reflux for three times.The ethanol extract was evaporated under reduced pressure to yield a residue(1634.2 g).And then,the residue was scattered in saturated water,partitioned with petroleum ether,dichloromethane,ethyl acetate and n-butanol.After evaporated the solvent,we got four portions of extracts with the weights of197 g,152 g,45.3 g and 165 g.Methods of silica gel column chromatography,Sephadex LH-20,polyamide column chromatography,preparative TLC,preparative HPLC and recrystallization were used in isolation and purification of the above four portions.Structures of compounds were elucidated by the spectral data of UV,MS,1H-NMR,13C-NMR,1H-1H COSY,HMQC,HMBC and NOESY.The antitumor activities of some compounds were evaluated by MTT colormetric method.Results: There were thirty compounds isolated from the aerial part of Achillea alpine L,and twenty-six compounds? structures were identified,including seven sesquiterpenoids,twelve flavonoids,two phenylpropanoids,two alkaloids and one fatty acid.The structures of these compounds were identified as follows:?-sitosterol(2),11?,13-dihydroparthenolide(3),11?,?13-?dihydroreynosin(4),11?,?13-?dihydrosantamarin(5),pectolinarigenin(6),p-hydroxycinnamic acid ester(7),6-(4?-hydroxy-1?,5?-dimethyl-5?-hexenyl)-3-methyl-2-cyclohexenone(9),compressanolid(10),Artemitin(12),1,?10-?epoxydihydropartheno?lide(13),9,?16-?dioxo-?10,?12,?14-?octadecatrienoic acid(14),tricin(15),eupatorin(16),centaureidin(17),artapshin(18),?-daucosterol(19),apigenin(20),apigenin-7-O-glucoside(21),hispidulin(22),2-hydroxy-4(1H)-quinolinone(23),2,?3,?4,?9-?tetrahydro-?1H-?pyrido[3,?4?b]?indole-?3-?carboxylic acid(24),chlorogenic acid(25),Luteolin-7-O-glucoside(26),isorhamnetin-3-O-rutinoside(27),rutin(28),tricin-7-O-glucoside(30).In vitro antitumor assay showed that tricin(15),apigenin(20),hispidulin(22)exhibited strong cell growth inhibitory activity against the human gastric cancer cell line SGC-7901,GES-1,human lung cancer cell line A549 and NCI-H292.While pectolinarigenin(6)and centaureidin(17)exhibited moderate antinproliferative activity against human cancer cell lines tested above.Conclusions: Feasible and reasonable methods have been carried out to investigate the constituents of different polar portions of the aerial part of Achillea alpina L.And twenty-six compounds were identified,among them,the bisabolane sesquiterpene 6-(4?-hydroxy-1?,5?-dimethyl-5?-hexenyl)-3-methyl-2-cyclohexenone is a new compound firstly found in nature;Guaiane-type sesquiterpenoid: compressanolide(10),9,?16-?dioxo-?10,?12,?14-?octadecatrienoic acid(14),tricin(15)and alkaloids: 2-hydroxy-4(1H)-quinolinone(23),2,?3,?4,?9-?tetrahydro-?1H-?pyrido[3,?4?b]?indole-?3-?carbox ylic acid(24)are isolated for the first time from the genus Achillea;Germacrane-type sesquiterpenoids 11?,13-dihydroparthenolide(3),1,?10-?epoxydihydropartheno?lide(13),eudesmane-type sesquiterpenoids1?,?13-?dihydroreynosin(4),11?,?13-?dihydrosantamarin(5),artapshin(18),flavonoids pectolinarigenin(6),centaureidin(17),apigenin(20),hispidulin(22)are isolated from Achillea alpina L for the first time.In addition,compounds tricin(15),apigenin(20)and hispidulin(22)exhibited strong antitumor activity. |
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