Study On The Chemical Constituents Of The Rhizome Of Kaempferia Galanga L.

Aromatic ginger（Kaempferia galanga L.）,belongs to the family Zingiberaceae,which is mainly distributed in the tropical and subtropical Asia including China,Myanmar,Indonesia,Malaysia,and Thailand.In China,it has been widely cultivated in Taiwan,Guangdong,Guangxi and Yunnan.In traditional Chinese medicine,the rhizome of aromatic ginger is prescribed for relieving pain and promoting digestion.The volatile oil extracted from the rhizome of K.galanga could be widely used in food,cosmetics,and medicines.According to modern pharmacological studies,the rhizome of K.galanga has been reported possessed broad bioactivities,such as antitumor,anti-inflammation,antioxidant,and antipyretic activities.At present,the chemical constituents of K.galanga are rarely reported,including flavonoids,terpenoids,and volatile oil.In this study,the rhizomes of K.galanga were extracted with 75%ethanol-water,and the extracts were successively extracted with cyclohexane,chloroform,ethyl acetate,and n-butanol,respectively.Various chromatographies were applied to the separation and purification of the constituents.Meanwhile,on basis of the physical and chemical properties and spectroscopic data（1D and 2D NMR,MS）,51 compounds were successfully isolated and elucidated.They were identified as cyclic dipeptides:Cyclo-（L-Val-L-Phe）（1）,Cyclo-（L-Leu-L-Ile）（2）,Cyclo-（L-Val-L-Leu）（3）,Cyclo-（L-Val-L-Val）（4）,Cyclo-（L-Ala-L-Ile）（5）,Cyclo-（L-Ala-L-Leu）（6）,Cyclo-（L-Ala-L-Phe）（7）,Cyclo-（L-Val-L-Ala）（8）,Cyclo-（L-Phe-L-Tyr）（9）,Cyclo-（L-Leu-L-Tyr）（10）,Cyclo-（L-Val-L-Tyr）（11）,Cyclo-（L-Asp-OCH₃-L-Phe）（12）,Cyclo-（L-Tyr-L-Ile）（13）,Cyclo-（L-Pro-L-Tyr）（14）,Cyclo-（L-Leu-L-Phe）（15）,Cyclo-（L-Glu-OCH₃-L-Phe）（16）;pyroglutamyl dipeptides:L-pGlu-L-Leu-OCH₃（17）,Pyroglutamyl-phenylalanine methyl ester（18）,Pyroglutamyl-tyrosinemethylester（19）;diarylheptanoids:（1R,3R,5R）-1,5-Epoxy-3-hydroxy-1-（3,4-dihydroxyphenyl）-7-（3,4-dihydroxyphenyl）heptane（20*）,（1R,3R,5R）-1,5-Epoxy-3-hydroxy-1-（3,4-dihydroxyphenyl）-7-（4-hydroxy phenyl）heptane3-O-β-D-glucopyranoside（21*）,1-（4-hydroxy-3-methoxyphenyl）-7-（4-hydroxyphenyl）heptane-1,2,3,5,6-pentaol（22*）,Phaeoheptanoxide（23）,（3R,5S）-3,5-Dihydroxy-1,7-bis（3,4-dihydroxyphenyl）heptane（24）,HedycoropyranB（25）;terpenoids:6β-Acetoxy-1α-14α-dihydroxyisopimara-8（9）,15-diene（26*）,Hedytriol（27）,（3R,4R,6S）-3,6-Dihydroxy-1-menthene（28）,Oxyphyllenodiol B（29）,（1R,2S,4R）-p-Menth-5-ene-1,2,8-triol（30）;phenolics:1-O-4-Carboxylphenyl-（6-O-4-hydroxybenzoyl）-β-D-glucopyranoside（31*）,trans-4-Methoxy ethyl cinnamate（32）,Ferulic acid（33）,trans-p-Hydroxy-cinnamic acid（34）,trans-4-Methoxycinnamic acid（35）,Methyl（2R,3S）-2,3-dihydroxy-3-（4-methoxyphenyl）propanoate（36）,Ethyl（2R,3S）-2,3-dihydroxy-3-（4-methoxyphenyl）propanoate（37）,p-Hydroxybenzoic acid（38）,p-Methoxybenzoic acid（39）,Vanillic acid（40）,Methyl3,4-dihydroxybenzoate（41）,4-methoxybenzyl-O-β-D-glucopyranoside（42）;other types:Linolenic acid（43）,Linoleic acid（44）,5,6-Dimethyl citrate（45）,3-Carboxyethyl-3-hydroxyglutaric acid 1,5-dimethyl ester（46）,Trimethyl citrate（47）,1,5-Dimethyl citrate（48）,Furan-2-carboxylic acid（49）,β-Sitosterol（50）,β-Daucosterol（51）.Among these purified constituents,compounds 1-16 are sixteen cyclic dipeptides,compounds 17-19 are three pyroglutamyl dipeptides,compounds 20-25 are six diarylheptanoids,compounds 26-30 are five terpenoids,compounds 31-42 are phenolics,and compounds 43-51 are belong to other types.Among them,compounds 20-22、26、31 are five new compounds.Diarylheptanoids were evaluated for their inhibitory effects on lipopolysaccharide（LPS）-induced nitric oxide（NO）production in a macrophage RAW 264.7 cell line.Except for compound 25,most of diarylheptanoids(IC₅₀ 17.26-79.89μmol/L)exhibited pronounced inhibitory activities compared to indomethacin(IC₅₀ 32.26±1.76μmol/L).The results demonstrated that diarylheptanoids isolated from the rhizome of K.galanga show significant anti-inflammatory effect.