Studies On The Chemical Constituents Of The Stems Of Tinospora Sinensis And Their Bioactivities

Tinospora sinensis belongs to the genus Tinospora of the family Menispermaceae,and is widely distributed and cultivated in the South of China.The stems of this species,commonly known as "Qing-Jiu-Niu" in Chinese,have been used as a traditional Yao medicine to treat rheumatism,lumbar muscle strain,and sciatica.Previous phytochemical studies on T.sinensis have led to the isolation and identification of diterpenoids,lignans,and several other constituents.The crude extracts of T.sinensis exhibited diverse biological activities such as cytotoxic and anti-inflammatory effects.The stems of T.sinensis were investigated on their chemical constituents systematically by various kinds of chromatographic methods.Fourty-eight compounds were obtained from EtOAc fraction.These compounds were identified as follows:tinosinenosides A-D(1-4),4-epi-tinosinenoside D(5),tinosinenosides E-F(6-7),2-deacetyltinosinenoside D(8),4-epi-2-deacetyltinosinenoside D(9),2-deacetoxytinosinenoside D(10),4-epi-2-deacetoxytinosinenoside D(11),tinosinenoside G(12),tinosineside A(13),1-deacetyltinosposide A(14),tinosineside B(15),tinosinenosides H-I(16-17),tinocordiside(18),betulalbuside A(19),vervenone-10-O-β-D-glucopyranoside(20),tinosinenosides J-M(21-24),seco-isolariciresinol 9-O-β-glucopyranoside(25),sinapyl alcohol 9-0-(E)-p-coumaroyl-4-O-β-D-glucopyanoside(26),(+)-pinoresinol 4-O-β-D-glucopyranoside(27),(+)-Syringaresinol(28),tangeoside A(29),meliasendanin C(30),(-)-isolariciresinol 3a-O-β-D-glucopyranoside(31),vladinol D(32),trans-N-p-coumroyl tyramine(33),N-trans-feruloyltyramine(34),N-trans-caffeoyltyramine(35),(E)-3-[(2,3-trans)-2-(4-Hydroxy-3-methoxyphenyl)-3-hydroxymethyl-2,3-dihydrobenzo[b][1,4]dioxin-6-yl]-N-(4-hydroxyphenethyl)acrylamide(36),thoreliamide B(37),grossamide K(38),cis-grossamide K(39),daidzin(40),syringin(41),tinosiene(42),trans-ferulic acid(43),vanillic acid(44),benzyl β-D-glucoside(45),1H-indole-3-carbaldehyde(46),4-[2-formyl-5-(methoxymethyl)-1H-pyrrol-1-yl]butanoic acid(47),5-hydroxymethylfuraldehyde(48).Among the isolated fourty-eight compounds,there were fifteen diterpenoids(1-15),three sesquiterpenoids(16-18),two moniterpenoids(19-20),nineteen phenylpropanoids(21-39)and nine other components(40-48).Compounds 1-12 and compounds 16-17 were new terpenoids,compounds 21-22 were new lignans,and compounds 23-24 were new phenylpropionic acids.The major components isolated from T.sinensis were clerodane diterpenoids,which possessed multiple chiral centers.The relative configurations of compounds 1-12 were determined by comprehensive analysis of 1D and 2D NMR data,and their absolute configurations were further established by using experimental and calculated ECD data.Compounds 1-18,21-24 were assessed in terms of their cytotoxic effects against HeLa cells.Only 1-deacetyltinosposide A(14)showed mild cytotoxicity,with an IC50 value of 8.35 ±0.60 μM;no signs of cytotoxicity were found for the other compounds(IC50 ≥ 10μM).Additionally,the NO production inhibitory activity of compounds 1-15 on LPS-activated N9 microglial cells was also tested;the results showed that none of these compounds had inhibitory activity(IC50 ≥ 100μM).