小葉三點(diǎn)金的黃酮類成分研究

劉小輝,王長虹,侴桂新,華 燕

1西南林學(xué)院,昆明 650224; 2中藥標(biāo)準(zhǔn)化教育部重點(diǎn)實(shí)驗(yàn)室上海中醫(yī)藥大學(xué)中藥研究所,上海 201210

小葉三點(diǎn)金的黃酮類成分研究

劉小輝1,2,王長虹2,侴桂新2,華 燕1＊

1西南林學(xué)院,昆明 650224;2中藥標(biāo)準(zhǔn)化教育部重點(diǎn)實(shí)驗(yàn)室上海中醫(yī)藥大學(xué)中藥研究所,上海 201210

從小葉三點(diǎn)金 (Desm odium m icrophyllum)乙醇溶液中分離得到 9個(gè)黃酮類化合物,通過波譜學(xué)方法分別鑒定為 (–)-表兒茶素 (1)、genistein-4′-O-β-glucoside(2)、異葒草苷 (isoorientin,3)、葒草苷 (orientin,4)、木犀草素 (luteotin,5)、異日本獐牙菜素 (isos wertiajaponin,6)、luteoayamenin(7)、木犀草素-7-O-β-D-葡萄糖苷 (luteolin-7-galactoside,8)、木犀草素-7-O-鼠李糖苷 (luteolin-7-rhamnoside,9)。所有化合物均為首次在該植物中分離得到。

小葉三點(diǎn)金;化學(xué)成分;黃酮

小葉三點(diǎn)金 (Desm odium m icrophyllum)為豆科山螞蝗屬 (Desm odium)植物,別名斑鳩窩、辮子草、碎米柴、爬地香、哮靈草、小葉山綠豆等,以其根及全草入藥。有清熱解毒、利濕通絡(luò)、消炎止血、活血化瘀的功能,在民間用于治療尿路感染、糖尿病、泌尿系結(jié)石、慢性胃炎、慢性氣管炎等癥[1,2]。近年來對山螞蝗屬的植物研究發(fā)現(xiàn),該屬植物具有良好的鎮(zhèn)痛、消炎、抗氧化、保護(hù)血管等作用[3-6]。我們對民間藥用植物小葉三點(diǎn)金進(jìn)行了系統(tǒng)的化學(xué)成分研究,從中分離得到 9個(gè)黃酮類型的化合物,均為首次在該植物中分離得到。

1 儀器和材料

LCQ DECAXPplus質(zhì)譜儀;Brucker AM500型核磁共振儀(T MS為內(nèi)標(biāo));柱層析硅膠和薄層層析硅膠(青島海洋化工廠);凝膠 Sephadex LH-20(Pharmacia公司)。

小葉三點(diǎn)金采自云南省嵩明縣,由西南林學(xué)院杜凡教授鑒定為小葉三點(diǎn)金 Desm odium m icrophyllum(Thunb.)DC.。

2 提取與分離

小葉三點(diǎn)金藥材全草 5 kg用 75%乙醇提取三次,濾液濃縮得到流浸膏約 2.4 L,大孔吸附樹脂(D101型)吸附,然后依次用水、35%、65%、95%乙醇沖洗,其中 35%乙醇段為黃酮段 (212 g)。黃酮段先用硅膠柱色譜,以氯仿 -甲醇梯度洗脫,由10∶1洗脫部分得到化合物 1(17 mg)、2(28 mg);由 4∶1洗脫部分得到化合物 3(5 mg)、4(20 mg)、6(8 mg)、7(10 mg);由 1∶1洗脫部分得到化合物 5(13 mg)、8(9 mg)和 9(7 mg)。

3 結(jié)構(gòu)鑒定

化合物 1 黃褐色晶體 (甲醇水),C15H14O6。 mp.310～312 ℃,[α= -55.39°(c0.31, MeOH);ESI-MSm/z:289[M-H]–;1H NMR(500 MHz,CD3OD)δ:6.97(1 H,d,J=1.9 Hz,H-2′), 6.79(1 H,dd,J=8.2,1.9 Hz,H-6′),6.75(1 H,d, J=8.2 Hz,H-5′),5.94(1 H,d,J=2.3 Hz,H-8), 5.91(1 H,d,J=2.3 Hz,H-6),4.52(1 H,d,J= 8.0 Hz,H-2),4.17(1 H,m,H-3),2.85(1 H,dd,J =16.8,4.6 Hz,H-4α),2.73(1 H,dd,J=16.8,2.9 Hz,H-4β)。13C NMR(125 MHz,CD3OD)δ:80.17 (C-2),67.78(C-3),29.54(C-4),157.96(C-5), 96.71(C-6),158.29(C-7),96.20(C-8),157.66 (C-9),100.39(C-10),132.59(C-1′),116.20(C-2′),146.23(C-3′),146.07(C-4′),115.62(C-5′), 119.71(C-6′)。以上數(shù)據(jù)與文獻(xiàn)[7]報(bào)道一致,故確定該化合物為(–)-表兒茶素。

化合物 2 淡黃色晶體 (甲醇),C21H20O10。mp.255～257℃;ESI-MSm/z:477[M+HCOO]–;1H NMR(500 MHz,DMSO-d6)δ:12.93(1 H,s,5-OH),9.60(1 H,s,7-OH),8.42(1 H,s,H-2),7.40 (2 H,d,J=8.5 Hz,H-2′,6′),6.83(2 H,d,J=8.5 Hz,H-3′,5′),6.72(1 H,d,J=2.2 Hz,H-8),6.47 (1 H,d,J=2.2 Hz,H-6),5.06(1 H,d,J=7.8 Hz,H-1″),3.71(1 H,m,H-6″a),3.51(2 H,m,H-5″,6″b),3.28(2 H,m,H-2″,3″),3.18(1 H,m,H-4″)。13C NMR(125 MHz,DMSO-d6)δ:154.55(C-2),122.54(C-3),180.49(C-4),161.61(C-5), 99.55(C-6),162.99(C-7),94.51(C-8),157.19 (C-9),106.06(C-10),120.97(C-1′),130.13(C-2′,6′),115.06(C-3′,5′),157.47(C-4′),99.84 (C-1″),73.05(C-2″),76.38(C-3″),69.58(C-4″),77.16(C-5″),60.60(C-6″)。以上數(shù)據(jù)與文獻(xiàn)[8]報(bào)道一致,故確定該化合物為 genistein-4′-O-βglucoside。

化合物 3 亮黃色粉末 (甲醇),C21H20O11。mp.255～258℃;ESI-MSm/z:447[M-H]–;1H NMR(500 MHz,DMSO-d6)δ:13.57(1 H,s,5-OH),10.56(1 H,br.s,7-OH),9.91(1 H,s,4′-OH),9.41(1 H,s,3′-OH),7.43(1 H,dd,J=2.2, 8.2 Hz,H-6′),7.41(1 H,d,J=2.2 Hz,H-2′),6.90 (1 H,d,J=8.2 Hz,H-5′),4.60(1 H,d,J=9.8 Hz,H-1″),糖上的其它氫信號δ:4.86～3.20 ppm。13C NMR(125 MHz,DMSO-d6)δ:164.33(C-2), 103.50(C-3),182.56(C-4),161.38(C-5),94.17 (C-6),163.91(C-7),109.55(C-8),156.88(C-9),104.10(C-10),122.12(C-1′),113.99(C-2′), 146.43(C-3′),150.37 (C-4′),116.74 (C-5′), 119.67(C-6′),73.72(C-1″),71.30(C-2″),79.62 (C-3″),70.88(C-4″),82.27(C-5″),62.18(C-6″)。以上數(shù)據(jù)與文獻(xiàn)[9]報(bào)道一致,故確定該化合物為異葒草苷。

化合物 4 黃色粉末 (甲醇),C21H20O11。mp.256～258℃;ESI-MSm/z:447[M-H]-;1H NMR (500 MHz,DMSO-d6)δ:13.17(1 H,s,5-OH), 10.80(1 H,br.s,4′-OH),10.02(1 H,s,3′-OH), 9.04(1 H,s,7-OH),7.53(1 H,dd,J=1.8,8.4 Hz,H-6′),7.48(1 H,d,J=1.8 Hz,H-2′),6.87(1 H,d,J=8.4 Hz,H-5′),糖區(qū)還給出氫信號δ:4.97～3.26 ppm。13C NMR (125 MHz,DMSO-d6)δ: 164.08 (C-2),102.41 (C-3),182.02 (C-4), 160.37(C-5),98.11(C-6),162.53(C-7),104.53 (C-8),155.99(C-9),104.03(C-10),122.02(C-1′),114.08(C-2′),145.80(C-3′),149.58(C-4′), (C-5′),119.37(C-6′),78.76(C-1″),73.39(C-2″),70.88(C-3″),70.69(C-4″),82.00(C-5″), 61.63(C-6″)。以上數(shù)據(jù)與文獻(xiàn)[10]報(bào)道一致,故確定該化合物為葒草苷。

化合物 5 土黃色粉末 (氯仿甲醇),C15H10O6。mp.328～330℃;ESI-MSm/z:285[M-H]-;1H NMR (500 MHz,CD3OD)δ:7.37(1 H,d,J=8.7 Hz,H-6′),7.37(1 H,br.s,H-2′),6.89(1 H,d,J=8.7 Hz,H-5′),6.53(1 H,s,H-3),6.43(1 H,d,J=1.3 Hz,H-8),6.20(1 H,d,J=1.5 Hz,H-6)。13C NMR (125 MHz,CD3OD)δ:164.82(C-2),102.68(C-3),182.69(C-4),162.04(C-5),98.91(C-6), 165.18(C-7),93.79(C-8),158.23(C-9),104.12 (C-10),122.50(C-1′),112.96(C-2′),145.86(C-3′),149.80(C-4′),115.59(C-5′),119.10(C-6′)。以上數(shù)據(jù)與文獻(xiàn)[011]報(bào)道一致,故確定該化合物為木犀草素。

化合物 6 亮黃色粉末 C22H22O11。mp.280～282℃;ESI-MSm/z:461[M-H]-;1H NMR (500 MHz,DMSO-d6)δ:13.50(1 H,br.s,5-OH),9.96 (1 H,br.s,4′-OH),9.35(1 H,br.s,3′-OH)為活潑氫信號;7.48(1 H,br.s,2′-H);7.47(1 H,d,J= 8.0 Hz,6′-H),6.91(1 H,d,J=8.0 Hz,5′-H),6.79 (1 H,s,3-H),6.78(H,s,6-H),3.90(3 H,s,6-OCH3)。13C NMR(125 MHz,DMSO-d6)δ:164.02 (C-2),103.09(C-3),182.12(C-4),159.56(C-5),90.04(C-6),163.72(C-7),104.57(C-8), 156.78(C-9),104.57 (C-10),121.34 (C-1′), 113.50(C-2′),145.76 (C-3′),149.79 (C-4′), 115.95 (C-5′),119.03 (C-6′),72.80 (C-1″), 70.82(C-2″),79.06(C-3″),70.25(C-4″),81.65 (C-5″),61.73(C-6″),56.45(6-OCH3)。以上數(shù)據(jù)與文獻(xiàn)[12]報(bào)道一致,故確定該化合物為異日本獐牙菜素。

化合物 7 亮黃色粉末,C22H22O11。mp.184～185℃;ESI-MSm/z:461[M-H]-;1H NMR (500 MHz,DMSO-d6)δ:13.50(1 H,br.s,5-OH),9.96 (1 H,br.s,4′-OH),9.35(1 H,br.s,3′-OH)為活潑氫信號;7.48(1 H,br.s,2′-H),7.47(1 H,d,J= 8.0 Hz,6′-H),6.91(1 H,d,J=8.0 Hz,5′-H),6.75 (1 H,s,3-H),6.73(1 H,s,8-H),3.87(3 H,s,6-OCH3)。13C NMR(125 MHz,DMSO-d6)δ:163.88 (C-2),103.09(C-3),181.80(C-4),160.30(C-5),109.64(C-6),164.92(C-7),90.88(C-8), 156.78(C-9),104.07 (C-10),121.34 (C-1′), 113.50(C-2′),145.76 (C-3′),149.79 (C-4′), 115.95 (C-5′),119.03 (C-6′),72.55 (C-1″), 70.90(C-2″),79.06(C-3″),70.25(C-4″),81.83 (C-5″),61.73(C-6″),56.21(6-OCH3)。以上數(shù)據(jù)與文獻(xiàn)[13]報(bào)道一致,故確定該化合物為 Luteoayamenin。

化合物 8 亮黃色粉末 C21H20O11。mp.230～232℃;ESI-MSm/z:447[M-H]-;1H NMR (500 MHz,DMSO-d6)δ:13.03(1 H,br.s,5-OH),9.61 (2 H,br.s,3′,4′-OH),為活潑氫信號。13C NMR (125 MHz,DMSO-d6)δ:165.02(C-2),103.73(C-3),182.42(C-4),161.68(C-5),98.57(C-6), 163.06(C-7),95.17(C-8),157.02(C-8a),105.91 (C-4a),121.90(C-1′),114.06(C-2′),146.33(C-3′),150.48(C-4′),116.53(C-5′),119.71(C-6′), 100.99(C-1″),74.52(C-2″),79.85(C-3″),70.95 (C-4″),77.52(C-5″),61.02(C-6″)。以上數(shù)據(jù)與文獻(xiàn)[14]報(bào)道一致,故確定該化合物為木犀草素-7-O-β-D-葡萄糖苷。

化合物 9 亮黃色粉末,C21H20O10。mp.233～234℃;ESI-MSm/z:431[M-H]-;1H NMR (500 MHz,DMSO-d6)δ:13.03(1 H,br.s,5-OH),9.61 (2 H,br.s,3′,4′-OH),為活潑氫信號。13C NMR (125 MHz,DMSO-d6)δ:165.02(C-2),103.73(C-3),182.42(C-4),161.68(C-5),98.57(C-6), 163.06(C-7),95.17(C-8),157.02(C-9),105.91 (C-10),121.90(C-1′),114.06(C-2′),146.33(C-3′),150.48(C-4′),116.53(C-5′),119.71(C-6′), 99.83(C-1″),70.30(C-2″),72.39(C-3″),64.73 (C-4″),68.87(C-5″),18.64(C-6″)。以上數(shù)據(jù)與文獻(xiàn)[15]報(bào)道一致,故確定該化合物為木犀草素-7-O-鼠李糖苷。

1 Jiangsu NewMedical College(江蘇新醫(yī)學(xué)院),Zhong Yao Da CiDian(中藥大辭典),Shanghai:Shanghai Science and Technology Press,1977,2466-2467.

2 Wu ZY(吳征鎰),Xin Hua Ben Cao Gang Yao(新華本草綱要),Shanghai:Shanghai Science and Technology Press, 1991,130.

3 Govindarajan R,VijayakumarM,Rao CV.Antiinflammatory and Antioxidant Activities ofDesmodium gangeticum Fractions in Carrageenan-induced Inflamed Rats.Phytochem istry, 2007,21:975-979.

4 Giuliano Delle Monache,Bruno Botta,Vittorio Vinciguerra. Antimicrobial isoflavanones fromDesm odium canum.Phytochem istry,1996,41:537-544.

5 Anshu Rathi,RAO CV,Ravishankar B.Anti-inflammatory and anti-nocieptive activity of thewater decoctionDesm odium gangeticum.Ethnophar m acology,2004,95:259-263.

6 Gino A.Kurian,Sachu P,Varghee T.Effect of aqueous extract of theDesm odium gangeticumDC root in the severity of myocardial ingfarction.Ethnophar m acology,2005,97:457-461.

7 Xin WB(辛文波),Chou GX(侴桂新),Wang ZT(王崢濤). Chemical constituents in the leaves ofUncaria hirsute Haviland.Chin J NatMed(中國天然藥物),2008,64:262-263.

8 Philip TL,Kristiina W?h?l?.Regiospecific 4′-O-β-glucosidation of isoflavones.Tetrahedron Lett,1998,39:9559-9562.

9 SunWX(孫武興),Li X(李銑),LiN(李寧),et al.Chemical constituentsof the extraction of bamboo leaves fromPhyllostachys nigra (Lodd.ex Lindl.)Munro var.henonis (Mitf.)Stepf.ex Rendle,J Shenyang Phar m lUniv(沈陽藥科大學(xué)學(xué)報(bào)),2008,25:39-41.

10 SongDM(宋冬梅),Sun QS(孫啟時(shí)).Chemical studies on the constituents ofTrollius alticusC.A.Mey.Chin J M ed Chem(中國藥物化學(xué)雜志),2004,14(4):233-234.

11 FengWS(馮衛(wèi)生),Li HW(李紅偉),Zheng XK(鄭曉珂),et al.Chemical constituents from the leaves ofB roussonetia papyrifera.Acta Phar maceut Sin,2008,43:173-180.

12 Marina Della Greca,Maria Ferrara,Antonio Fiorentino.Antialgal compounds fromZantedeschia aethiopica.Phytochem istry,1998,49:1299-1304.

13 Ryosuke Hirose,Yukio Kazuta,Daizo Koga,et al.On the Structure of C-Glycosyl Flavones,Flavoayamenin and Luteoayamenin,in Petals ofIris NertshinskiaLoddiges form Albiflora Honda.Agric B iol Chem,1981,45:551-555.

14 Zhong H(仲浩),Xue XX(薛曉霞),Yao QQ(姚慶強(qiáng)). Studies on chemical constituents ofScutellaria barbata.Chin Tradit Herb D rugs(中草藥),2008,39:21-23.

15 Fayez E Kandil,Mary H.Grace.Polyphenols fromCornulaca monacantha.Phytochem istry,2001,58:611-613.

Flavonoids ofDesm odium m icrophyllum

L IU Xiao-hui1,2,WANG Chang-hong2,CHOU Gui-xin2,HUA Yan1＊1Southwest Forestry College,Kunm ing 650224,China;2Key Laboratory of Standardization of ChineseM edicines,M inistry of Education,Institute of ChineseM aterialM edic of Shanghai University of Traditional ChineseM edicine,Shanghai 201210,China

Nine flavonoidswere isolated from thewhole plantofDesmodium m icrophyllum,and identified as(–)-epicatechin(1),genistein-4′-O-β-glucoside(2),isoorientin(3),orientin(4),luteotin(5),isos wertiajaponin(6),luteoayamenin(7),luteolin-7-galactoside(8),and luteolin-7-rhamnoside(9).These compoundswere isolated fromDesm odium m icrophyllumfor the first time.

Desmodium m icrophyllum;chemical constituents;flavonoids

1001-6880(2010)06-0976-04

2008-12-15 接受日期:2009-04-09

中藥標(biāo)準(zhǔn)化教育部重點(diǎn)實(shí)驗(yàn)室開放課題基金資助(ZK-0601)

＊通訊作者 Tel:86-871-3863022;E-mail:huayan@mail.kib.ac.cn

R284.1;Q946.91

A