Study On The Chemical Constituents And Activity Of Miao Medicine Jiuzilianhuancao

Objective:To study on the chemical constituents and activities of Calanthe discolor and find with novel structure and positive activity compounds.It provides the research basis literature for rational development and application of the plant.Method:1.The whole plants of Calanthe discolor were extracted with methanol three times(7 days/time).The total extract was extracted with petroleum ether,dichloromethane,ethyl acetate and n-butanol respectively.The extract isolated and purified by silica gel column chromatography,aluminium column chromatography,polyamide column chromatography,Sephadex chromatography,C18,MCI and recrystallization.The isolated compounds were identified by 1H-NMR,13C-NMR,MS,X-Ray single crystal diffraction,physicochemical properties and related databases.2.The evaluation of different parts were tested with MTT methods on antineoplastic activities,and the activation of macrophages by different sites were tested by ELISA methods.Mycobacterium tuberculosis H37Rv was tested by drug-based methods in vitro.Results:1.35 compounds were isolated from Calanthe discolor and identified as squalene(1),,?-sitosterol(2),ergosta-7,22-dien-3?-ol(3),22E-5?,8?-epidioxyergosta-6,22-dien-3?-ol(4),stigmast-4,6,8(14)-22-tetraen-3-one(5),24-methylenecycloartan-ol(6),3,5-dimethoxyphenanthrene-2,7-diol(7),indirubin(8),1H-imidazole-4-carboxylate(9),methyl anthranilate(10),4-hydroxyphenylpropionic methyl ester(11),methyl ferulate(12),methyl trans ferulate(13),mehyl trans coumaric acid(14),3,3'-dihydroxy-5-methoxybibenzyl(15),1-isobutyl-4-(2'-ethyl-n-hexyl)-terephthalate(16),N-benzoyl-L-phenylalaninol(17),pyrocatechol(18),caffeic acid methyl ester(19),p-coumaric(20),p-hydroxy-phenylpropionic acid(21),meth-yl 4-hydroxy-3,5-dimethoxy cinnamate(22),methyl salicylate(23),dibutyl phth-alate(24),1,2-benzendicar-boxylic acid bis[2-methylpropyl]ester(25),bis(2-ethylhexyl)benzene-1,2-dicarboxylate(26),wrightiadione(27),pinostrobin(28),xanthone(29),quercetin-3-O-?-D-galactopyranoside(30),sucrose(31),maltose(32),methyl hexadecanoate(33),n-hexadecane(34),dotriacont-l-ene(35).2.Compared with the positive drug doxorubicin,(1)the petroleum ether part(QY-1)has a weak inhibitory effect on Hela;(2)the water part(QY-5)has a weak inhibitory effect on BEL-7402;(3)all sites have no inhibitory effect on BGC-823;(4)the petroleum ether part(QY-1)and dichloromethane part(QY-2))have a certain inhibitory effect on A549,the ethyl acetate site(QY-3)has a weak inhibitory effect on A549;(5)the n-butanol part(QY-4),water part(QY-5)and total extract(QY-6)have a weak inhibitory effect on T-24;(6)the petroleum ether part(QY-1)has weak inhibitory effect on K562,total extract has a weak inhibitory effect on K562?3.The ethyl acetate fraction(QY-3)has a certain activation effect on macrophages;The ethyl acetate fraction(QY-3)has effect on inhibited H37Rv.Conclusion:1.Among the 35 compounds isolated from Calanthe discolor,there are 17 phenolic acid compounds,5 steroids,4 flavonoids,2 alkaloids,2 carbohydrate,1 phenanthrene,1 triterpenes and 3 aliphatics.Thirty-three of the above compounds were isolated from this plant for the first time.2.All parts showed certain activities in anti-tumor,which provided a scientific basis for folk using the plant to treat a variety of cancers.QY-1 and QY-3 parts played a more extensive role.Especially the QY-3 part,had an effect of inhibiting H37Rv,which suggested the rationality of widely using the plant in treatmenting cervical lymphatic tuberculosis.