印度蛇菰的三萜成分

印度蛇菰的三萜成分 3 印度蛇菰的三萜成分 刘锡葵李忠荣邱明华聂瑞麟 ( ) 中国科学院昆明植物研究所 , 昆明 650204 ( ( ) ) 摘要 从我国民间药用植物印度蛇菰 Arn. Griff . 中分离得到 7 个化合 B al anop hora i n dica ((β) ) β物 , 经鉴定为 : 棕榈酰- 香树酯 蛇菰素 A、棕榈酰羽扇豆烯醇酯 蛇菰素 B、乙酰- 香树 β酯 、乙酰羽扇豆烯醇酯 、- 香树脂酮 、羽扇豆烯酮及棕榈酸 。运用光谱和化学的方法对它们的 结构进行解析 。其中羽扇豆型萜为首次自该植物中分得 ; 蛇菰素 A 和 B 具有较强的护肝作用 。 关键词 印度蛇菰 , 三萜酯 , 蛇菰素 A , 蛇菰素 B 分类号 Q 946 Triterpene Const ituents f rom B a l a nop hor a i n dica L IU Xi - Kui L I Zho ng - Ro ng Q IU Ming - Hua N I E Rui - Lin ( )L aboratory of Phytoche m ist ry , Ku n m i n g I nst i t ute of B ot a ny , T he Chi nese A ca de m y of S ciences , Kunming 650204 β Abstract Fro m herbs of t he Chinese fol k medical plant B al anop hora i n dica , seven co mpounds : - ( ) ( ) β amyrin palmitate balanop horin A, lupeol palmitate balanop horin B, - amyrin acetate , lupeol ac2 β etate , - amyro ne , lupeo ne and palmitic acid were isolated.Their st ruct ures were elucidated by t he ba2 sis of spect ral and chemical evidences , respectively. The co mpounds of lupane were obtained f ro m B . i n dica , firstly. Balanop horin A and B exhibited st ro ng activit y against liver damage induced by CCl. 4 Key words B al anop hora i n dica , Triterpene ester , Balanop horin A and B ()B al anop hora i n dica Arn. Griff . was a kind of parasitic plant wit h root , belo ng to species of t he family Balanop horaceae . It is used as a fol k medicine plant for t he to nics and hemostatic , indigenous to t he p rovince Yun2 nan . To our knowledge , t he biologically active p rinciple and co nstit uent s f ro m t his plant , have not been described as yet . Therefore we st udied o n t he co nstit uent s of it . Here we report o n t he investigatio n of t he isolatio n and st ruct ural elucidatio n of t wo t riterpene esters f ro m t his plant . RES UL TS A ND D ISCUSSIO N ( ) The pet roleum benzine ext ract of t he herbs of B al anop hora i n dica Arn. Griff , was subjected to repeated β( column chro matograp hy o n silica gel and aluminum o xide , to give 7 co mpounds : - amyrin 3 - palmitate 1 , ) balanop horin A, β β( ) ( ) ( ) ( ) - amyro ne 2 ,- amyrin 3 - acetate 3 ,lupeol 3 - palmitate 4 , balanop horinB , lupeo ne 5 , lupeol 3 - 3 中国科学院昆明植物研究所植物化学开放实验室基金资助 1997 - 09 - 17 收稿 , 1998 - 04 - 21 接受发表 () () acetate 6and palmitic acid 7. The yield of balanop horin A and B were 1121 % and 1134 % , respectively. - 1 ( ) Balanop horin A showed a molecular io n peak at m/ z 665cm M + 1 ,CHOin t he mass spect rum. The IR 46 80 2 - 1 spect rum showed at 1710 ,1165 ,and haven’t at 3300,3500cm ,indicating t he p resence of a carbo nylo xy group 1 () δ( ) O = C - O. The H - NM R spect rum appeared o ne olefinic p roto n signal at 5115 1 H ,brs,o ne o xygen - bear2 δδ() (ing carbo n p roto n signal at4147 1 H ,br ,t J = 714 Hzin t he lowfield range ;and nine met hyl p roto n at1111 3 H , ) ( ) ( ) ( ) ( ) δ s, 0194 3 H , s, 0 . 93 3 H , s, 0 . 84 15 H , brs , 5 ×CH, 0 . 80 3 H , s; seventeen met hylene p roto ns at 1 . 23 3 () δ34 H ,brs ,17 ×CH,and t he great majorit y p roto ns signal showed gat her at0 . 80 - 2 . 28 rang in t he highfield. 2 It suggested t hat balanop horin A could be an t ritepene ester derivative. The IR spect rum absorp tio n at 1370 ,1350 , β( were indicated t hat balanop horin A could be belo ng to t he- amyrin t ype t riterpene ester derivative Snatzke et al 13 δ) () ,1962. The C NM R and D EP T spect rum of balanop horin A revealed o ne carbo nyl carbo n 173 . 4,nine met hyl (δδ) ( carbo ns 24 . 3 ,16 . 8 ,25 . 9 ,28 . 0 ,32 . 4 ,23 . 7 ,14 . 1, six quaternary carbo ns 38 . 2 ,39 . 8 ,37 . 1 ,41 . 7 ,32 . 6 , ) δδ( ) ( 34 . 8,four met hine carbo n 80 . 5 bearing an o xygen ato m,55 . 2 ,47 . 5 ,47 . 2 and t wo loefinic carbo ns 121 . ) β 6d ,145 . 1s. These data can be acco mmodated o n t he- amyrin t ype t riterpene having lo ng chain fat ty acid. TheEI - mass spect rum of balanop horin A showed a characteristic f ragment io n for t he loss of palmitic acid at m/ z 218 13 1 ( ) βbase peak. On co mpariso n of t he C NM R and H - NM R spect rum of balanop horin A wit h t hat of- amyrin (β) () Bhat tacharyya et al ,1986,was identified as- amyrin 3 - palmitate 1. The EI - mass spect rum of t he al ka2 βline hydrolysis p roduct was correspo nding wit h t hat of- amyrin ,and f urt her co nfirmed t hat balanop horin A should βbe assigned to t he- amyrin ester . Fro m t he above evidence ,t he st ruct ure of balanop houin A was established to be β- amyrin 3 - palmitate. 13 Balanop horin B was calculated for CHOby t he FABMS and C NM R spect ro met ry. The IR spect rum ap2 46 80 2 1 ( ) ( ) peared t wo absorp tio n at 1710 C = O,and 1625 C = C. The H NM R spect rum was quite similar to t hat of bal2 () () δanop horin A ,but it showed t wo olefinic p roto n signals at 4 . 66 1 H ,s,4 . 54 1 H ,s;eight met hyl p roto n at0 . 76 () () ( ) ( ) ( ) ( ) 3 ,s,0 . 81 9 H ,s , 3 ×CH, 0 . 84 3 H , s, 0 . 92 3 H , s, 1 . 00 3 H , s, 1 . 66 3 H , s; and seventeen met hylene 3 13 δ) δ (() p roto ns at1 . 23 34 H ,brs ,17 ×CH. The C NM R and D EP T spect rum revealed o ne carbo n 173 . 5, t wo 2 (δδ ) () olefinic carbo ns 109 . 4t ,150 . 8s,five quaternary carbo ns 38 . 9 , 4 . 09 , 38 . 1 , 42 . 7 , 43 . 0, and six met hine (δ) carbo n 80 . 7 ,55 . 5 ,50 . 4 ,37 . 2 ,48 . 4 ,48 . 0. It exp ressed t hat balabop horin B could be a lupeol t ype derivative. ( The EI - mass spect rum of it exhibited a characteristic f ragment io n for t he loss of plamitic acid at m/ z 409 M - ) () 256,239 ,257 ,amd 426 . It was identified as lupeol 3 - palmitate 4by co mpariso n of t he data of NM R wit h lupe2 () ol Dreyer et al ,1972. Al kaline hydrolysis of it yields a t riterpene co mpound. The EIMS of t he t riterpene was cor2 respo nding wit h t hat of lupeol . The st ruct ure of it was established to be lupeol 3 - palmitate ,again. This co mpound ( ) was obtained f ro m B . i n dica , firstly Yadagiri et al ,1984 ; Chengalur et al ,1976. (Balanop horin A and B exhibited st ro ng activities against liver damage induced by CClin mice Lin et al , 4 ) 1988. EXPERIM ENTAL General procedures The NM R spect ra were performed choroformed using TMS as int . standard at 400M Hz wit h a Brucker AM - 400 inst rument . The carbo n type was determined by D EP T experiment s. IR spect ra were recorded in KBr pellet s o n a Per kin - Elmer 577 interfero meter . EIMS and FABMS :positive , direct inlet 70eV o n ( ) ( V G Autospec inst rument . For CC , silica gel 200 ,300 mesh , Qingdao and aluminum o xide neut ral , 200 ,300 ) () mesh ,Shanghai. TL C p recoated silic gel plate HF0 . 25mm in t hickness. 254 () Plant material The whole plant of B al anop hora i n dica Arn. Griff were collected in Xishuangbanna , Yun2 nan p rovince ,in October ,1996 ,and identified by Mr . Cui J ing - yun , Xishuangbanna Tropical Botanic Garden , t he Chinese Academy of Scinences ,where t he herbarium specimen has been deposited. Identif ication of the kno wn triterpenes All of t he known were identified by co mpariso n wit h aut hentic sam2 ples by t heir NM R or MS spect ra data . () Extraction and Separation of triterpenes The dry whole plant powered material 700 gwas ext racted wit h () pet roleum benzine yielding ,af ter evap n ,a brown yellow oil residue 80 g. The pet roleum benzine ext ract was dis2 solved in benzene and ext racted wit h met hanol . The benzene layer ,o n evap n of t he solvent , 46 g residue was ob2 (tained. The benzene ext ract chro matograp hied o n a column of silica gel wit h pet roleum benzine - aceto ne 20?1,2 ) ( ) ( ) ?1to give 3 f ractio ns in increasing of polarit y. Fractio n 2 29 gwas p urified by CC o n aluminum o xide neut ral () () () wit h benzene - met hanol 4?1to f urnish balanop horin A 8 . 5 gand B 9 . 4 g. Fractio n 3 was separated similarly ββ( ) ( ) ( as t hat for f ractio n 2 to afford- amyrin 3 - acetate 40 mg, - amyro ne 21 mg. And lupeol 3 - acetate 34 ) () mg,lupeo ne 25 mg. Palmitic acid was obtained f ro m f ractoin 1 . () ( ) Balanop horin A 1. Amorp hous white powders MeCO,mp 77 ?,dissolved in chloroform ,pet roleum ben22 KBr- 1 ν( ) zine and benaene. IRcm ?2900 ,2825 ,1710 ,1450 ,1370 ,1350 ,1235 ,1165 ,1085 ,980 ,710 . FABMSm/ z?max 1 ) ( ) ( ) ( ) ( δ665 M + 1. EIMS m/ z?665 ,409 ,218 base peak,257 ,239 ,203 ,190 ,175 ,109 ,69 . H NM R CDCl?5 . 15 3 () () ( ) ) ( 1 H ,brs ,12 - H,4 . 47 1 H ,t ,J = 7 . 4 Hz ,3 - H,2 . 26 2 H ,t ,J = 7 . 4 Hz ,2’- H,1 . 23 34 H , brm , CH,1 . 11 2 () ( ) ( ) ( ) ( 3 H ,s , 27 - CH, 0 . 94 3 H , s , 30 - CH, 0 . 93 3 H , s , 29 - CH, 0 . 84 15 H , brs , CH, 0 . 80 3 H , s , 16’- 3 3 3 3 13 ) () (CH. C NM R data see table. Balanop horin A 20 mg,was t reated wit h NaO H - HO 210 g NaO H dissolved in 3 2 ) 20 ml HOat 160 ?for 12 hr . The reactio n mix was ext racted wit h CHCl,af ter evap n ,obtained a residue. The 2 3 ( ) EIMS spect ra of t he residue exhibited a f ragment io n peak at m/ z 426 ,218 base peak. KBr - 1 () ( ν) Balanop horin B 4. Amorp hous white powders MeCO, mp 68 ,69 ?. IRcm ?3040 , 2900 , 2825 , max 2 ( ) ( ) 1710 ,1625 ,1450 ,1440 ,1438 ,1370 ,1165 ,970 ,900 ,710 . EIMS m/ z?665 M + 1,409 ,426 ,257 ,239 ,218 ,95 ,1 ( ) ) ( δ ( ) ( ) ( 68 ,55 base peak. H NM R CDCl?4 . 46 1 H , s , 29 - H, 4 . 54 1 H , s , 29 - H, 4 . 44 1 H , dd ,J = 10 . 5 ,3 ) ( ) ( ) ( ) ) (516 Hz ,3 - H,1 . 23 34 H ,brm ,CH,1 . 66 3 H , s ,30 - CH,1 . 00 3 H , s ,27 - CH,0 . 92 3 H ,24 - CH,0 . 2 3 3 3 () () () () ( ) 84 3 H ,s ,29 - CH,0 . 81 9 H ,s,0 . 76 3 H ,s,0 . 76 3 H ,s ,16’- CH,2 . 26 2 H ,t ,J = 716 Hz ,2’- CH,2 . 31 3 3 2 13 () ( ) ( 1 H ,m ,18 - H. C NM R data see table. Balanop horin B 20 mg,was t reated wit h NaO H - HO 2 . 0g NaO H 2 ) dissolved in 20ml HOat 160 ?for 12 hr . The EIMS spect ra of t he p roduct exhibited a f ragment io n peak at m/ 2 z 426 ,189 ,203 . 1 13 β() ( ) - Amyro ne 2. CHO , M424 . Colorless needles MeO H. The EIMS , H and C - NM R spect ra data 30 48 β( ) were identical to t he p ublished reference spect ra data of- amyro ne Go nzalez , 1981.et al 1 β() ( ) - Amyrin acetate 3. CHO,M468 . Colorless pillars MeCO,mp 236,237 ?Similary ,t he EIMS H32 52 2 2 13 β( and C - NM R spect ra data were identical to t hose of t he reference co mpound ,yrin acetate Seo - amet al ) 1975. 1 13 () ( ) L upeno ne 5. CHO M424 . White needles MeO H. The EI - MS , H and C - NM R spect ra were cor2 30 48 , () respo nding wit h t hat of lupene 3 - o ne Budzi kiewicz et al , 1963. 13 表 1 蛇菰素 A 和 B 的 C NMR 数据13Table 1 C NMR data of balanop ho rin A and B C Balanop ho rin A Balanop ho rin B C Balanop ho rin A Balanop ho rin B ( ))( 蛇菰素 A 蛇菰素 B 蛇菰素 A 蛇菰素 B 碳序 碳序 3717 t 18 t 19 t 14 t 1 3721 3129 2316 t 2314 t 3618 t 4010 t 2 22 8015 t 8017 t 2814 q 2715 q 3 23 3812 s 38154 s 1618 q 1613 q 4 24 5512 d 5515 d 1515 q 1613 q 5 25 1812 t 1813 t 1618 q 1613 q 6 26 3313 t 3413 t 2518 q 1415 q 7 27 3918 s 4019 s 2810 q 1813 q 8 28 4715 d 5014 d 3214 q 10914 t 9 29 3711 s 3811 s 2317 q 1914 q 10 30 2315 t 2110 t 17314 s 17315 s 11 ′1 12116 d 2512 t 3419 t 3418 t 12 2′ 14511 s 3712 d 2511 t 2512 t 13 3′ 14 4117 s 4217 s 2913 t 2913 t 4′ 15 2619 t 2715 t ???? ? ? 16 2612 t 3516 t 13′ 2917 t 3018 t 17 3216 s 4310 s 14′3110 t 3119 t 18 4712 d 4814 d 15′2217 t 2217 t 19 4618 t 4810 d 20 3418 s 15018 s 16′ 1411 q 1415 q Measured in CDCl,pp m. 3 () ( ) L upeol acetate 6,CHO,M468 . Colorless pillars MeO H,mp 217,218 ?. The EIMS and NM R spec2 32 52 2 ( ) t ra data were identical to t hose of t he reference co mpound lupeol 3 - acetate Wenkert et al , 1978. ( ) ( ) Palmitic acid 7. CHO, M256 . Colorless powders MeO H. The EIMS and NM R spect ra were corre2 16 32 2 ( ) spo nding wit h t hat of palmitic acid Vanderlan et al , 1991. Ackno wledgements We are gratef ul to t he Inst rument Analysis Group of t he Phytochemist ry Depart ment , Kunming Instit ute of Botany for measurement s of all spect ra . REFERENCES ( ) Bhat tacharyya J ,Barro s C B ,1986 . Triterpenoids of , 25 1?274,276 Cni doscul us u rens . Phytoche m ist ry Budzikiewicz H , Wilso n J M , Djerassi C , 1963 . Mass spect ro met ry in st ruct ural and stereochemical p ro blems XXXI I pentacyclic ( ) t riterpenes. J A mer Che m S oc , 85 22?3688,3699 Chengalur R N ,Asho k K K ,Chelat Q et al , 1976 . Chemical examinatio n of t he r hizo mes of Coffee Fungus , B al a nop hora i n dica ( ) Wall . ,14B 11?906,907I n di a n J Chem S ect B Dreyer D L , Pickering M V ,Co hn P ,1972 . Dist ributio n of limo noids in t he Rutaceae . , 11Phytoche m ist ry ?705,71313( )Go nzalez A G , Fraga B M , Go nzalez P et al , 1981 . C - NMR spect ra of olean - 18 - ene derivatives. Phytoche m ist ry , 20 8 ?1919,1921 ( ) Lin C N , To me W P ,1988 . Antihepatoto xic p rinciples of , 54 3?223,224S a m bucus f or m osa na . Pl a nt Medici ne Seo S , To mita Y , To ri K ,1975 . Carbo n - 13 NMR Spect ra of U RS - 12 ene applicatio n to st ruct ural assignment s of co mpo nent s ( ) of Isodon j aponicus Hara tissue cult ures. Tet ra L et t , 16 1?7,10 Snatzke G ,L amper F , Tschesche R , 1962 , U ber Triterpene ? Zuo rdnung Vo n t riterpene Zu Den Grund t ypen Durch IR Spek2 t ro skoipe. Tet rahed ron , 18?1417,1431 Vanderlan D S B , Ligia M V T , Claudia M M Y , 1991 . Caffic acid esters and t riterpenes of A l i bert i a m acrophyl l a . ( ) Phytoche m ist ry , 30 6?2089,2091 Wenkert E ,Baddeley G V ,Burfit t I R et al , 1978 . Carbo n - 13 Nuclear magnetic reso nance spect ro scop y of nat urally - occurring substance L ?. t riterpene related to lupene and hopane . , 11 O rg M a ng n Res ?337,343Yadagiri B K R ,Subba G S R R , 1984 . Triterpenio ds f ro m , 47 ( ) B al a nop hora i n dica . J N at Prod 1?1823 3 3 3 3 3 3 3 3 3 3 被引频次最高的中国科技期刊 500 名排列表中 云南植物研究名列第三十八位 中国生物学类科技期刊排行表 ()按被引频次和影响因子排序 名次 期刊名称 被引频次名次 期刊名称 影响因子 植物生态学报 0145561 1 植物学报 943 2 植物学报 0142862 植物生理学通讯 553 3 遗传学报 0139263 植物生理学报 489 4 植物生理学报 0138604 遗传学报 419 5 古脊椎动物学报 0132655 生物化学与生物物理进展 383 6 细胞生物学杂志 0128576 植物分类学报 366 7 微生物学报 0125497 生物化学与生物物理学报 323 8 植物分类学报 0125008 云南植物研究 308 9 人类学学报 0124449 古植物学报 266 10 实验生物学报 01239710 动物学报 264 11 数据来源 :中国科学院文献情报中心中国科学引文数据库 1996 年数据。本表由中国科学引文数据库统计编制。 21 被引频次是对被中国科学引文数据库 1996 年 582 种来源期刊所引用的数千种中国出版的中英文期刊进行频次统计后 编制而成的。 年引用 1994 年和 1995 年该刊刊载总次数 1996 31 影响因子的计算方法如下 :1996 年某刊的影响因子 = 1994 年和 1995 年该刊刊载论文的总篇数 本表中 1996 年的影响因子是在对中国科学引文数据库 1994,1995 年的来源期刊作了统计后编制而成。41 本着尊重原始数据的原则 ,本表对变名期刊未作任何合并处理。