Studies On The Constituents And Biological Activities Of Clematis Manshurica

Clematis manshurica Rupr.,belonging to Ranunculaceae family,is mainly distributed in northeast,Shanxi,Shandong,Hebei,eastern part of Inner Mongolia of China and is also widely distributed in Korea and Russia.It is commonly used as a traditional Chinese medicine "Wei Ling Xian",which has the effect of dispelling wind,removing dampness and unblocking collaterals to alleviate pain,and as an ethno-medicine commonly used for the treatment of rheumatism and arthritis.The modern pharmacology studies have shown that this plant showed strong anti-inflammatory and anti-tumor activities.We reviewed the chemical constituents and pharmacological activities of Clematis genus.The roots and rhizomes of C.manshurica were subjected to phytochemical investigation.The antiproliferative effects of some isolated compounds against PC-3 human prostate cancer cells in vitro were evaluated.By means of various chromatographic methods such as D101 macroporous resin,opening silica gel,Sephadex LH-20 column chromatography and preparative HPLC,75 compounds were isolated from the 60%EtOH extracts of the roots and rhizomes of C.manshurica,and their structures were elucidated on the basis of spectroscopic data and physico-chemical methods,such as ID,2D NMR and HR-ESI-MS.These compounds include 30 triterpenoid saponins,14 phenolic glycosides,1 lignan,1 stilbene,8 lactones,4 fatty acids,2 saccharides,3 sterols,10 alkaloids and amines.12 new compounds were named as clematomanshurica saponins F(1),G(2),H(3),I(4),J(5),K(6),L(17),clemomandshuricoside D(31),manshuricaphenside A(33),manshuricaterside A(48),2-(2-hydroxyethyl amino)ethyl icosanoate(73),and 2-(2-hydroxyethyl amino)ethyl linoleate(75).6 new natural products were 2-(2-hydroxyethyl amino)ethyl octanoate(68),2-(2-hydroxyethyl amino)ethyl decanoate(69),2-(2-hydroxyethyl amino)ethyl dodecanoate(70),2-(2-hydroxyethyl amino)ethyl tetradecanoate(71),2-(2-hydroxyethyl amino)ethyl hexadecanoate(72),and 2-(2-hydroxyethyl amino)ethyl icosanoate(74).The other known compounds were identified as raddeanoside 15(7),hederacolchiside E(8),huzhangoside B(9),cernuoside A(10),saponin PJ3(11),3-O-β-D-glucopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1-2)[β-D-glucopyranosyl-(1→4)]-α-L-arabinopyranosyl oleanolic acid(12),CP7a(13),CP7(14),CP4(15),CP2(16),kizutasaponin K10(18),kizutasaponin K12(19),leontoside D(20),ciwujianoside A(21),hederacolchiside F(22),3-O-a-L-rhamnopyranosyl-(1→3)-a-L-rhamnopyranosyl-(1→4)-a-L-arabinopyranosyl hederagenin 28-0-a-L-rhamnopyranosyl-(1→4)-β-D-glucopyranosyl-(1→6)-β-D-glucopyranoside(23),3-O-a-L-rhamnopyranosyl-(1→3)-α-L-rhamnopyranosyl-(1→4)-α-L-arabinopyranosyl hederagenin 28-0-β-D-glucopyranosyl-(1→6)-β-D-glucopyranoside(24),cernuoside A(25),oleanolic acid(26),21a-hydroxyoleanolic acid(27),21β-hydroxyoleanolic acid(28),corosolic acid(29),tormentic acid(30),5-O-feruloyl-3-O-(β-D-glucopyranosyl)-2-deoxy-D-ribono-y-lactone(32),5-0-feruloyl-3-O-2-deoxy-D-ribono-y-lactone(34),ferulic acid(35),methyl ferulate(36),ethyl ferulate(37),isoferulic acid(38),methyl isoferulate(39),methyl ferulic acid(40),methyl 3,4-dimethoxy cinnamate(41),1,2,4-trimethoxybenzene(42),3-hydroxy-4-methoxybenzoic acid(43),4-methoxybenzoic acid(44),asarinin(45),curcusinol(46),7-hydroxyterpineol(47),5R-5-hydroxymethyl-2(5H)-furanone(49),5R-dihydro-5-hydroxymethyl-2(3H)-furanone(50),5-hydroxymethyl-2-furaldehyde(51),3-hydroxy butanolide(52),2-hydroxymethyl-D-ribono-y-lactone(53),dihydro-3-hydroxy-5-hyroxymethyl-2(3H)-furanone(54),3-O-β-D-glucopyranosyl-2-hydroxymethyl-D-ribono-y-lactone(55),anemonin(56),5-hydroxy-4-oxopentanoic acid(57),methyl 3,4-dihydroxybutanoate(58),succinic acid(59),methyl docosanoate(60),L-rhamnose(61),glucose(62),β-stitosterol(63),stigmasterol(64),daucosterol(65),methyl pyroglutamate(66),and methyl 7-methoxy-3-indolecarbonate(67).Among them,compounds 7,8,10,12,23-25,28-30,36,37,39-41,45-47,60,66 were obtained from genus Clematis for the first time.Twenty-six isolated compounds were evaluated for antiproliferative activity against human prostate cancer PC-3 cell lines in vitro by MTT method.Compounds 5,6,12-17 exhibited strong cell growth inhibitory activities against PC-3 cell line,with the GI50 values of 1.50,1.29,0.71,1.25,1.32,2.07,3.10,2.62 μM,respectively.Of all these compounds,12 showed the maximum activity.