新疆紫草中抗生育化学成分_英文_
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天然产物研究与开发 Vo1.14 No.1NATURAL PRODUCT RESEARCH AND DEV ELOPMEN T A CONTRACEPTIVE CONSTITUENT FROM A RN EB IA EUC HRO MA (ROYL E)JO HNST 3.ZHAN G Hui 2zhen 1,2,L IAO Mao 2chuan 332,XUAN Li 2jiang 3,GUO Ji 2xian 1(1.School of Pharm acy ,Fudan U niversity ,Shanghai 200032,China ;2.N ational L aboratory of Drugs and Devices f or Family Planning ,S hanghai Institute of Planned Parenthood Research ,Shanghai 200032,China ;3.S hanghai Institute of M ateria Medica ,CA S ,Shanghai 200031,China )Abstract From the roots of A rnebia euchrom a (Royle )Johnst.,dipotassium rabdosiin 1,potassium rosmari 2nate 2,sodium rosmarinate 3and sodium ferulate 4,were isolated by means of reversed phase chromato 2graphy.Their structures were identified by spectral analyses (UV ,IR ,MS ,1H NMR ,13C NMR ).Com 2pound 1exhibited conspicuous contraceptive pound 2,3and 4were discovered in the roots of A.euchrom a firstly.K ey w ords A rnebia euchrom a (Royle )Johnst.;dipotassium rabdosiin ;contraceptive effect0 IntroductionThe root of A rnebia euchrom a is used as a traditional Chinese medicine named ”Xinjiang Zicao ”,which shows many pharmacological activities ,such as pro 2moting blood circulation ,counteracting toxicity and facilitating eruption.In 1982,Liu reported that the aqueous extract of A rnebia euchrom a could terminate the early pregnancy of mice and rabbits [1].In this pa 2per ,we reported the isolation and structural elucida 2tion of four phenolics :dipotassium rabdosiin 1,potas 2sium rosmarinate 2,sodium rosmarinate 3and sodium ferulate 4from the roots of A rnebia euchrom a .A 2mong them ,compound 1exhibited conspicuous con 2traceptive effect ,which could excite the uterine smooth muscle of mice and stimulate the necrosis of human decidual pound 2,3and 4were dis 2covered firstly from thisplant.收稿日期:2001211205 修回日期:20012122293The Project Supported by National Laboratory of Drugs and Devices for Family Planning.33Author to whom correspondence should be addressed.11 R esults and DiscussionCompound 1,a tan amorphous powder ,was separat 2ed by MCI gel CHP20P cc of the aqueous extract of the roots of A rnebia euchrom a ,and showed a dark 2blue coloration with FeCl 3reagent ,indicative of its phenolic nature.Its UV spectrum (H 2O )showed peaks at 252,282,316(sh )and 343nm.The IR spectrum revealed the presence of phenolic groups (3600~3300cm -1),conjugated carbonyl groups (1692cm -1),and aromatic rings (1600~1400cm -1).The 1H NMR (in D 2O )spectrum exhibited three one 2proton aromatic or olefinic singlets (δ6.52,6.87,7.53,s ,1H each ),two aliphatic methines (δ3.90,4.46,br.s ,1H each ),two oxygen 2bear 2ing methines (δ4.84,4.98,br.d ,1H each ),two methylenes (δ2.65~3.12,m ,4H ),and three cat 2echol rings.The protons at δ6.52and 6.87can be attributed to a 1,2,4,52tetrasubstituted benzene moiety ,and that at δ7.53to the β2proton of the caf 2feoyl group.The 13C NMR (in D 2O )spectrum showed signals due to four carboxyl carbons ,four aromatic rings ,a double bond ,two oxygen 2bearing carbons ,and four aliphatic carbons.These data wereidentical with those of rabdosiin [2] .Compound 1was suggested to be a salt because of its poor solubility in organic solvents.It was confirmed by IR spectral da 2ta.No signals above 1720cm -1were detected ,which are diagnastic to the caboxyl group.Instead ,signals at 1586cm -1and 1396cm -1were showed to be assigned to caboxylate moity.ESI 2MS showed the molecular ion peak at 795,which was 76more than that of robaosiin ,indicative of two potassium ion.Consistently ,atom absorbance spectrum confirmed the existence these two potassium ion quantatitively.Consequently ,compound 1was identified as dipotas 2sium rabdasiin.2 Experimental211 G eneralUV was tested by Lambda 20in H 2O.IR was recorded by 360F T 2IR Nicolet.NMR spectra were acquired on Bruker AM 2400,Bruker 2DRX 400and G emini 2300,with TMS as internal standard.EI 2MS data was obtained on HP5989A.Column chromatography was performed using MCI gel CHP20P (75~150μ,Mitsubishi Chemical In 2dustries Co.Ltd.),TSK gel Toyopearl HW 240F (30~60μ,TOSO Co.Ltd.),Cosmosil 75C 182OPN (42~105μ,Nacalai Tesque Inc.),Sephadex L H 220(25~100μ,Pharmacie Fine Chemical Co.Ltd.).TLC was conducted on precoated HSGF254plates (Qingdao Ocean Chemical Co.)with benzene 2ethyl acetate 2formic acid (2∶7∶1).212 Plant materialThe dried root of A rnebia euchrom a (Royle )Johnst.was bought from Shanghai Huayu Ltd.The voucher specimen was identified by Professor Guo Jixian and deposited in the Department of Pharmacognosy ,School of Pharmacy ,Fudan University.213 Extraction and IsolationThe dried roots of A rnebia euchrom a (8kg )were extracted with 70%aqueous Me 2CO.The aqueous Me 2CO extract was concentrated to yield an aqueous solution.After removal of the insolubles by filtration ,the filtrate was concentrated under reduced pressure ,and subjected to MCI gel CHP20P column chromato 2graphy with H 2O containing an increasing amount of MeOH to give five fractions :Fractions Ⅰ(196g ),Ⅱ(20.6g ),Ⅲ(22.3g ),Ⅳ(18.7g )and Ⅴ(10.5g ).Fraction Ⅰwas subsequently chromatographed over Cosmosil 75C 182OPN [H 2O 2MeOH (1∶0→3∶7)]to furnish compound 1(1.20g ).Repeated chro 22 天然产物研究与开发Vo1.14 No.1matography of fractionⅡon Sephadex L H220[H2O2 MeOH(1∶0→1∶1)],Cosmosil75C182OPN[H2O2 MeOH(1∶0→3∶7)],and TSK gel Toyopearl HW2 40F[H2O2MeOH(1:0→3:7)],afforded compounds 2(1.02g)and3(0.47g).FractionⅣwas chro2 matographed on Cosmosil75C182OPN[H2O2MeOH (1∶0→4∶6)]and then TSK gel Toyopearl HW240F[H2O2MeOH(1:0→1:1)]to yield compound4 (1.07g).214 Identif ication21411 Compound1,a tan amorphous powder. [α]22D2113.4°[c=0.91,H2O].UVλmax(H2O) nm:252,282,316(sh),343.IR(K Br)cm-1: 3600~3300(br.),1692,1586,1521,1396, 1242,1115,820,800.ESI2MS m/z:[M+K]+ 833,795,757.Elemental analysis found C 48.15%,H4.46%,calculated for C36H28O16K2・6H2O,C47.89%,H4.43%.1H NMR(300MHz, D2O)δ:2.65~3.12(m,H27”),3.90(br.s,H2 2),4.46(br.s,H21),4.84(br.dd,H28”),4.98 (br.dd,H28”),6.52(s,H28),6.87(s,H25), 7.53(s,H24),6.85(d,J=2Hz,H22”),6.76 (d,J=2Hz,H22”),6.75(d,J=8Hz,H25”), 6.67(d,J=8Hz,H25”),6.62(dd,J=2and8 Hz,H26”),6.37(dd,J=2and8Hz,H26”),6.61 (d,J=2Hz,H22’),6.64(d,J=8Hz,H25’), 6.34(dd,J=2and8Hz,H26’).13C NMR(100 MHz,D2O)δ:46.4(C21),48.4(C22),133.4(C2 3),142.4(C24),137.8(C24a),119.4(C25), 145.7(C26),149.5(C27),119.8(C28),126.2(C2 8a),123.2(C21’),117.6(C22’),145.3(C23’), 14612(C24’),118.6(C25’),122.1(C26’),132.7 (C21”),133.0(C21”),118.8(C22”),145.0(C2 3”),145.2(C23”),146.4(C24”),119.1(C25”), 12411(C26”),124.2(C26”),39.1(C27”),39.3 (C27”),79.6(C28”),79.8(C28”),179.3(C2 9”),179.7(C29”),176.4(22COO2),170.7(32 COO2).21412 Compound2,a tan amorphous powder.UVλmax(H2O)nm:197,220,284,324.IR(K Br) cm-1:3400~3000(br.),1692,1598,1521, 1447,1401,1263,1116,820,813.EI2MS m/z: 413[M2K]+.Elemental analysis found C48.58%, H4.74%,calculated for C18H15O8K・3H2O,C 47.79%,H4.65%.1H NMR(400MHz,D2O)δ: 7.08(s,H22),6.87(d,J=8Hz,H25),6.97(d, J=8Hz,H26),7.46(d,J=15.9Hz,H27),6.26 (d,J=16Hz,H28),5.06(dd,J=3.2and8.8 Hz,H210),3.01(dd,J=3.2and14.2Hz,H2 11),3.13(dd,J=8.8and14.2Hz,H211),6.91 (s,H213),6.87(d,J=7.8Hz,H216),6.80(d, J=7.8Hz,H217).13C NMR(100MHz,D2O)δ: 129.5(C21),117.7(C22),146.7(C23),149.5 (C24),118.8(C25),125.2(C26),148.6(C27), 117.7(C28),171.5(C29),79.0(C210),39.5(C2 11),132.8(C212),119.8(C213),146.4(C214), 145.3(C215),118.6(C216),124.4(C217), 179.7(C218).These data were identical with those of potassium rosmarinate[3,4].21413 Compound3,a white amorphous powder. UVλmax(H2O)nm:197,220,284,324.IR(K Br) cm-1:3400~3200(br.),1692,1598,1516, 1455,1396,1262,1115,820,800.ESI2MS m/z: [M+K]+421.Elemental analysis found C 50.72%,H4.88%,calculated for C18H15O8Na・3H2O,C49.54%,H4.82%.1H NMR(300MHz, D2O)δ:7.06(d,J=2Hz,H22),6.87(d,J=8 Hz,H25),6.95(dd,J=2and8Hz,H26),7.45 (d,J=16Hz,H27),6.25(d,J=16Hz,H28), 5.06(dd,J=4and9Hz,H210),3.01(dd,J=4 and14Hz,H211),3.13(dd,J=9and14Hz,H2 11),6.91(d,J=2Hz,H213),6.86(d,J=8 Hz,H216),6.80(dd,J=2and8Hz,H217).13C NMR(100MHz,D2O)δ:129.7(C21),117.1(C2 2),146.8(C23),149.6(C24),118.8(C25), 125.3(C26),148.6(C27),117.8(C28),171.6 (C29),79.1(C210),39.5(C211),132.9(C212), 119.9(C213),146.4(C214),145.3(C215), 118.8(C216),124.5(C217),179.8(C218).These data were identical with those of sodium rosmari2 nate[3,4].21413 Compound4,a white amorphous powder. UVλmax(H2O)nm:214,286,310.IR(K Br) cm-1:3500~3000(br.),1640,1596,1514, 1401,1275,1123,980,852,818.EIMS m/z: 194,179,150,135.1H NMR(400MHz,D2O)δ: 7.11(s,H22),7.02(d,J=8.1Hz,H25),6.84 (d,J=8.1Hz,H26),7.27(d,J=15.9Hz,H23Vo1.14 No.1张慧桢等:新疆紫草中抗生育化学成分 7),6.32(d ,J =15.9Hz ,H 28), 3.83(s ,32OCH 3).13C NMR (100MHz ,D 2O )δ:130.7(C 21),113.6(C 22),150.1(C 23),149.1(C 24),118.2(C 25),124.7(C 26),143.8(C 27),124.1(C 28),177.9(C 29),58.5(32OCH3).These data were identical with those of sodium ferulate [4].R eferences1 Liu F ,et al .Terminating the early pregnancy activity ofA rnebia euchrom a in mice and rabbits.Journal of Xinjiang Medical U niversity ,1982,5:1992 Isao A ,et al .A tetrameric derivative of caffeic acid fromRabdosia japonica.Phytochemist ry ,1989,28(9):24473 Charles J K ,et al .Carbon 213nuclear magnetic resonance oflithospermic and rosmarinic acids J Org Chem ,1975,40(12):18044 Charles J K ,et al .isoiation and.structure determination oflithospermic acid.J Org Chem ,1976,41(3):449新疆紫草中抗生育化学成分张慧桢1,2 廖矛川2 宣利江3 郭济贤1(11复旦大学药学院 上海 200032;2.上海市计划生育科学研究所国家计划生育药具重点实验室 上海 200032;3.中国科学院上海药物研究所 上海 200031)摘 要 从新疆紫草(A rnebia euchrom a (Royle )Johnst.)的根部分得四种酚类化合物,经波谱分析,确定为rabdosiin 二钾盐(dipotassium rabdosiin 1),迷迭香酸钾盐(potassium rosmarinate 2),迷迭香酸钠盐(sodium rosmarinate 3),阿魏酸钠盐(sodium ferulate 4)。