Study On The Bioactive Constituents Of The Flower Of Wedelia(sphagneticola Trilobata(L.)pruski)

Wedelia[Sphagneticola trilobata（L.）Pruski]is the genus Sphagneticola of the Compositae family,which is native to South America and is now widely distributed in the coastal areas of China,such as Guangdong,Fujian,Hainan,Hong Kong,and Taiwan.Wedelia had a significant allelopathic potential for neighboring native plants,which could inhibit the growth of other plants,and it is listed as one of the hundred worst invasive species around the world.Ethnopharmacogically,Wedelia was applied as a remedy for treating infectious,intractable wound,rheumatism,muscle cramps,sores and swellings in some rural communities.Modern pharmacological studies have shown that its extract has antioxidant,antitumor,anti-inflammatory,antibacterial,and hepatoprotective effects.In this paper,68 compounds were isolated and purified from the chloroform fraction of 80%ethanol extract of the flowers of S.trilobata by silica gel,Sephadex LH-20,ODS and preparative Rp-HPLC.Their structure were identified as:（1）Sesquiterpene compounds:1α-Acetoxy-4α-hydroxy-6α-isobutyryloxy-9β-15α-methylisovaleryloxyprostatolide（1*）,1β-Acetoxyl-4a-hydroxy-6β-isobutyryloxy-9a-angelinoyloxyprostatolide（2*）,1β-Acetoxy-4α-hydroxy-6β-isobutyryloxy-9β-15α-methylangelinoyloxyprostatolide（3*）,1a,9β-Diacetoxyl-4β-hydroxy-6a-methacryloxy-11β-methylprostatolide（4*）,1a,9β,4a-Trihydroxy-6β-isobutyryloxyprostatolide（5*）,1a,6a,9β-Trihydroxy-4,10a-dimethyl-5a,7a,8a-endesm-3-en-8,12-olide（6*）,1α,4β,8β-Trihydroxy-6α-isobutyryloxyeudesman-9,12-olide（7*）,1α,9β-Dihydroxy-4β-hydroxymethyl-6β-methacryloxy-13β-methyleudesmanolide（8*）,1α,4β,6α-Trihydroxy-8β-angeloyloxyprostatolide（9*）,1β-Acetoxy-4α,9α-dihydroxy-6β-acetoxyprostatolide（10*）,1a,9β-Dihydroxy-6a-isobutyroxy-4,10β-dimethyl-5a,7β,8β-endesm-3-en-8,12-olide（11*）,1β,9α-Diacetoxy-4α-hydroxy-6β-isobutyroxyprostatolide（12）,1β-Acetoxy-4α,9α-dihydroxy-6β-isobutyroxyprostatolide（13）,1β-Acetoxy-4α-hydroxy-6β-isobutyryloxy-9α-isovaleryloxyprostatolide（14）,1β-Acetoxy-4α,9α-dihydroxy-6β-methacryloxyprostatolide（15）,1β,9α-Diacetoxy-4α-hydroxy-6β-methacryloxyprostatolide（16）,Trilobolide-6-O-isobutyrate（17）,1β,9α-Diacetoxy-4α-hydroxy-6β-methacryloxyprostatolide（18）,Oxidoisotrilobolide-6-O-methacrylate（19）,Oxidoisotrilobolide-6-O-isobutyrate（20）,Oxidoisotrilobolide-6-O-isobutyrate（21）,1α,4β-Dihydroxy-6α-isobutyryloxy-8β-angeloyloxyprostatolide（22）.（2）Deterpene compounds:Kauradien-9（11）,16-oic acid（23）,Kaur-16-en-19-oic acid（24）,3α-Angeloyloxy-ent-kaur-16-en-19-oic acid（25）,3α-Tiglinoyloxy-ent-kaur-16-en-19-oic acid（26）,3α-Cimnamoyloxy-ent-kaur-16-en-19-oic acid（27）,ent-9α-Hydroxy-16-kauren-19-oic acid（28）,3α-Angeloyloxy-9β-hydroxyent-kaura-16-en-19-oic acid（29）,3α-Tiglinoyloxy-9β-hydroxyent-kaura-16-en-19-oic acid（30）,Tetrachyrin（31）;（3）Triterpenes:3β-O-palmitoyl-1β-amyrin（32）,3,4-seco-19（10→9）abeo-8α,9β,10α-eupha-4,24-dien-3-ol（33）,Glut-5-en-3β-ol（34）,3α-Acetoxy-27-hydroxylup-20（29）-en-28-oic acid methylester（35）.（4）Flavonoids:4?,6,7,8-Tetramethoxyfavones（36）,3?,8-Dimethoxyl-3,4?,6-trihydroxyquercetin（37）,3?,8-Dimethoxy-4?,6-dihydroxyflavone（38）,（2R）-5?,8-Dihydroxy-3?-methoxyflavanone（39）,Liquiritigenin（40）2,4,4?-Trihydroxychalcone（41）.（5）Other type compounds:1-[（2R,5R）-5-（（E）-5-Hydroxy-3-（hydroxymethyl）pent-3-en-1-yl）-4-methylenetetrahydrofuran-2-yl]propan-2-one（42*）,[（9R,3E,6Z）-8-Methyl-8-（（E）-4-oxopent-2-en-1-yl）-5,8-dihydro-2H-oxocin-3-yl]methyl acetate（43*）,[（9R,3E,6Z）-3-（hydroxymethyl）-8-methyl-5,8-dihydro-2H-oxocin-2-yl]pent-3-en-2-one（44*）（2R）-1-（4-（Hydroxymethyl）-3,6-dihydro-2H-pyran-2-yl）propan-2-one（45*）,4-（（5R）-5-（Hydroxymethyl）-2,5-dihydrofuran-3-yl）butan-2-ol（46*）,3-Indole formaldehyde（47）,8-Hydroxy-3-methoxy-5H-pyrido[2,1-c]pyrazin-5-one（48）,Methyl-（7S）-hydroxyhexadeca-8E,10Z,13Z-trienoate（49）,Stigmasterol（50）,Ergosta-7,24（24?）-dien-3β-ol（51）,Syringaresinol（52）,Mediolipin（53）,2,6,2?,6?-Tetramethoxy-4,4?-bis（2,3-epoxy-1-hydroxypropyl）biphenyl（54）,Syringaresinol-O-β-D-glucopyroside（55）,7-Methoxy-8-hydroxycoumarin（56）,Bis（2-ethylhexyl）phthalate（57）,Vanillin（58）,p-Hydroxyacetophenone（59）,7-Hydroxychromogen（60）,Amyl benzoate（61）,4-Allyl-2,6-dimethoxyphenol-β-D-glucopyranoside（62）,Ethyl linoleate（63）,Methyl heptadecanoate（64）,6,9-Heptadecadiene（65）,Linolenic acid（66）,Methyl linoleate（67）,Trilinolein（68）,respectively.Among them,there are35 terpenes,including 22 sesquiterpene lactones（1-22）;9 diterpenes（23-31）;4 triterpenes（32-35）,6 flavonoids（36-41）,and 27 other various type compounds（42-68）.Among them,compounds 1-11,42-46 were 16 new compounds.Furthermore,the anti-inflammatory and anti-tumor activities were evaluated for the compounds 1-8,12-31.The clonogenic survival and the cells morphology assays were carried out for compounds 1-3.The results exhibited that compounds 2(IC₅₀ 13.85±1.63μmol/L),16(IC₅₀ 7.06±1.39μmol/L),19(IC₅₀ 11.69±0.20μmol/L),20(IC₅₀ 9.67±0.31μmol/L)and 21(IC₅₀ 16.53±0.61μmol/L)showed strong antiproliferative activity against Hep G2 cells and incuced apoptosis.The colony numbers were dose-dependently reduced in HepG2 cells after treated with compounds 1-3.Meanwhile,compounds 6(IC₅₀ 11.77±0.83μmol/L)and 25(IC₅₀ 24.03±1.46μmol/L)showed strong NO inhibitory activity on BV-2 cells.