Studies On Chemical Constituents From Two Plants Lonicera Japonica And Viburnum Odoratissimum Of Caprifoliaceae And Their Biological Activities

Caprifoliaceae belongs to dicotyledonous plants,and it is an important and old family.Most of Caprifoliaceae plants are shrubs or vines.A total of twelve genera include Lonicera,Sambucus,Viburnum,Triosteum,Heptacodium,Symphoricarpos,Linnaea,Kolkwitzia,Abelia,Dipelta,Weigela and Leycesteria.There are about 800 kinds of plants in Caprifoliaceae,which are widely distributed all over the world,mainly in the eastern Asia and eastern North America.The Caprifoliaceae plants are beautiful and leafy,which are famous for their abundant ornamental plants,such as Viburnum macrocephalum,Abelia grandiflora and Weigelaflorida.In addition,many plants from Caprifoliaceae have some medicinal value,such as Lonicera japonica and Sambucus williamsii and so on.This paper researched on chemical composition and biological activities fom two plants of Caprifoliaceae(Lonicera japonica Thunb.and Viburnum odoratissimum Ker-Gawl.).The flower buds of L.japonica belong to the Lonicera genus in the Caprifoliaceae family,which have a long history and abundant resources.The efficacy of L.japonica is significant,and it is known as heat-clearing and detoxicating medicine since ancient times.It tastes sweet,cold in nature and attributive to the lung,heart and stomach meridians.It can clear heat,remove toxin,course wind and dissipate heat,which is mainly used in the treatment of carbuncle furuncle swollen,erysipelas,poisonous blood flow field,wind-heat cold,warm disease and fever.Now the researches on chemical components and pharmacological activities of L.japonica focused on flavonoids and coffee acyl quinic acid compounds.The pharmacodyamic material basis is not perfect.Thus,in order to clarify the pharmacodyamic material basis and provide new basis for the depth development and quality control of L.japonica,we carried out a systematic study on its chemical composition and biological activities.By the means of various chromatographic methods such as macroporous adsorption resin,silica gel,polyamide,ODS and reversed-phase preparative HPLC,eighty five compounds were isolated from the aqueous extract of the air dried flower buds of L.japonica.On the basis of analysis of the physical and chemical properties of compounds and combination with the MS,IR,OR,1D-NMR,2D-NMR and ECD methods,the chemical structures of all compounds were elucidated as:Lonjaponspiroside A(Al),Lonjaponspiroside B(A2),5-?-Carboxystrictosidin(A3),L-phenylalaninosecologanin B(A4),L-phenylalaninosecologanin(A5),(35,4R,5S)-4-Ethenyl-3-(?-D-glucopyranosyloxy)-4,4a,5,6,7,8-hexahydro-8-oxo-3H-pyrano[3,4-c]-pyridine-6-sulfonic acid(A6)?(E)-Aldosecologanin(A7),(Z)-Aldosecologanin(A8),Loniceranan A(A9),Loniceranan B(A10),Loniceranan C(All),Dimethyl-secologanoside Secoxyloganin(A12),Secoxyloganin(A13),Secologanin(A14),Secologanoside(A15),Secologanic acid(A16),Ethyl secologanoside(A17),Secologanin dimethy acetal(A18),Demethylsecologanol(A19),Sweroside(A20),6'-O-?-D-Apiofuranosylsweroside(A21),Vogeloside(A22),epi-Vogeloside(A23),Swertiamarin(A24),Loganic acid(A25),8-Epiloganic acid(A26),Loganin(A27),8-Epiloganin(A28),7-Ketologanin(A29),Harpagide(A30),Harpagoside(A31),6"-O-?-Glucopyranosylharpagoside(A32),Kingiside(A33),8-Epikingiside(A34),7S-O-Methylmorroniside(A35),Abscisic acid(A36),Bluemenol A(A37),Loniceracycloside(A38),Urceolide(A39),Betulalbuside A(A40),8-Hydroxygeraniol-8-O-?-D-glucoside(A41),Kankanoside P(A42),(2E,6Z)-2,6-Dimethyl-8-?-D-glucosyloxy-2,6-octadienoic acid(A43),Laminamplexoside C(A44),(Z)-2,6-Dimethyl-2-octene-1,8-diol(A45),Monoterpenoid acid(A46),Monoterpene(A47),3,4-Dicaffeoylquinic acid(A48),4,5-di-O-Caffeoylquinic acid(A49),5-Caffeoylquinic acid(A50),3-O-Caffeoylquinic acid(A51),3-O-Caffeoylquinic acid methyl ester(A52),Quercetin(A53),Isoquercitrin(A54),Quercetin 3-O-L-rhamnoside(A55),Rutin(A56),Tamarixetin 3-O-neohesperidoside(A57),Luteolin-7-O-glucoside(A58),Liquiritin(A59),Hesperidin(A60),(7'R,8'S)-Dihydrodehydrodiconiferyl alcohol 4'-O-?-D-glucopyranoside(A61),Icariside E4(A62),Icariglucoside(A63),(7R,8S)-erythro-7,9,9'-Trihydroxy-3,3'-dimethoxy-8-O-4'-neolignan-4-O-?-D-glucopyranoside(A64),(7R,8R)-threo-7,9,9'-Trihydroxy-3,3'-dimethoxy-8-O-4'-neolignan-4-O-?-D-glucopyranoside(A65),(7S,8R)-erythro-7,9,9'-Trihydroxy-3,3'-dimethoxy-8-O-4'-neolignan-4-O-?-D-glucopyranoside(A66),Eugenylglucoside(A67),Eugenyl-?-D-xylopyranosyl-(1?6)-?-D-glucopyranoside(A68),Eugenol rutinoside(A69),4-Allyl-2-methoxyphenyl-6-O-?-D-apiosyl-(1?6)-?-D-glucoside(A70),Hesperidin(A71),?-D-Xylopyranosyl-(1"?6 ')-?-D-glucopyranoside(A72),?-D-Glucopyranose 6-[(2E)-3-phenylprop-2-enoate](A73),?-D-Glucopyranose 6-[(2E)-3-phenylprop-2-enoate](A74),Dihydroconiferylalcohol(A75),Caffeic acid(A76),trans-Cinnamic acid(A77),4-Methoxyphenyl-acetaldehyde(A78),4-Hydroxybenzaldehyde(A79),(3R)-1-Octen-3-yl?-D-primereroside(A80),Icariside G1(A81),Lonfuranacid A(A82),Lonfuranacid B(A83),trans-4-Hydroxy-2-nonenoic acid(A84),4-(2-Formyl-5-hydroxymethylpyrrol-1-yl)butyric acid(A85),including six monoterpene indole alkaloids,twenty nine iridoid glycosides,two sesquiterpenoids,ten monoterpenoids,five coffee acyl quinic acids,eight flavonoids,six lignans and nineteen others.Of all these isolates,eight compounds(A1,A2,A9,A10,A11,A63,A82,A83)were new,thirty two compounds were isolated from Lonicera genus for the first time.In order to further research the biological activities of all kinds of ingredients from L.japonica,we tested the anti-inflammatory effects of twenty eight iridoid glycoside compounds(A7-A20,A22-A35)isolated from L.japonica,based on the evaluation of the inhibitory effect on NO production in lipopolysaccharide(LPS)-induced RAW 264.7 cells.We observed that compounds A7 and A8 exhibited strong NO inhibitory activity with IC50 values of 12.60±1.50 and 7.96± 0.47?M,respectively,while the other compounds did not display any significant activity.In addition,compounds A7 and A8 had no effects on the survival rates of RAW 264.7 cells at a concentration of 100 ?M.We tested the anti-diabetic effects of fifty two compounds(A1-A20,A22-A35,A53,A55-A62,A64-A65,A67-A68,A70-A72,A82-A83)isolated from L.japonica,based on the evaluation of their inhibitory effect on two therapeutic targets(?-glucosidase and PTP1B).It was notable that compounds A1 and A2,displayed significant anti-diabetic activities in vitro involving both ?-glucosidase and PTP1B inhibition with IC50 values of 0.81± 0.04 and 0.74 ± 0.02 mM;6.14±0.53 and 8.42 ± 0.64?M,respectively.In order to clarify the mode of action of compounds A1 and A2 on ?-glucosidase and PTP1B,the molecular docking studies were carried out to measure the relative binding energies and localize binding sites in them.The molecular docking results were in good agreement with the in vitro inhibitory assay.Viburnum odoratissimum belongs to the Viburnum genus in the Caprifoliaceae family.Its variants are named Viburnum odoratissimum var.Sessiliflorum(Geddes)Fukuoka and Viburnum odoratissimum var.Awabuki(K.Koch)Zabel ex Rumpl,respectively.Its branches and leaves could be used as medicine with clearing heat and damp,activating the channels and collaterals and detoxify.V.odoratissimum is mainly used in the treatment of rheumatic pain,swelling pain and fractures.In addition,it has anesthetic effect,which is reversible.Modern pharmacological studies have shown that V.odoratissimum exhibits a broad spectrum of biological activities including anti-tumor,antibacterial and ichthyism effects.Now dozens of compounds including diterpenoids,triterpenoids,sesquiterpenoids,lignans and coumarins were isolated and identified from V.odoratissimum.V.odoratissimum mainly contains vibsane type diterpenoid compounds.They exists only in V.odoratissimum and Viburnum awabuki,which are limited in the nature.In addition,vibsane type diterpenoid compounds has complicated structures and significant anti-tumor activity.Thus,in order to research the vibsane type diterpenoid compounds with significant anti-tumor activity,we carried out a systematic study on its chemical composition and biological activities.First,we tested the cytotoxic activities of different macroporous adsorption resins(water,15%EtOH,60%EtOH,95%EtOH)against HepG2,Hep3B and U251 cell lines.We observed that the 95%EtOH effluent fraction displayed significant activities against HepG2 with IC50 values of 11.54±0.64 ?g/mL,against Hep3B with IC50 values of 17.26± 0.93?g/mL and against U251 with IC50 values of 36.26±2.41 ?g/mL.According to the cytotoxic activity results,we carried a guide separation.By the means of various chromatographic methods such as silica gel,CHP20,sephadex LH-20,ODS,normal-phase preparative HPLC,reversed-phase preparative HPLC,thirty one compounds were isolated from the 95%EtOH effluent fraction of V.odoratissimum.On the basis of analysis of the physical and chemical properties of compounds and combination with the MS,OR,1D-NMR and 2D-NMR methods,the chemical structures of all compounds were elucidated as:Vibsanum A(B1),Vibsanin A(B2),Vibsanol C(B3),Vibsanine B(B4),Vibsanin L(B5),Vibsaniguration A(B6),Vibsaniguration B(B7),Vibsanin H(B8),Vibsanin K(B9),Vibsanin G(B10),Vibsansuspenside A(B11),Vibsansuspenside B(B12),20-Hydroxy-3-O-oxolup-28-oic acid(B13),23-Hydroxy-3-oxo-urs-12-en-28-oic acid(B14),Cyperenol acid(B15),Spathulenol(B16),Loliolide(B17),Menthiafolic acid(B18),Vibsanlignan A(B19),Secoisolariciresinol(B20),Cycloolivil(B21),Dihydrodehydodiconiferyl alcohol(B22),Vibsanlignan B(B23),(+)-Pinoresinol(B24),5-Hydroxy-3,4-dimethyl-5-pentylfuran-2(5H)-one(B25),Phthalic acid isodibutyl ester(B26),Dibutyl phthalate(B27),4-Methoxycinnamic acid(B28),p-Hydroxycinnamic acid(B29),Caffeic acid(B30),3,4,5-Trimethoxy galic acid(B31),including eleven vibsane type diterpenoids,two iridoid glycosides,two triterpenoids,two sesquiterpenoids,two monoterpenoids,six lignans and seven others.Of all these compounds,seven compounds(B1,B6,B7,Bll,B12,B19,B23)were new,seven compounds were isolated from Viburnum genus for the first time.Then we tested the cytotoxic activities of vibsane type diterpenoid compounds(B1-B11)isolated from the 95%EtOH effluent fraction against HepG2,A549 and SH-SY5Y cell lines.The results showed that most compounds displayed moderate to strong activities.