黃圣卓,蔣思萍,朱華結

1中國科學院昆明植物研究所植物化學與植物資源持續(xù)利用國家重點實驗室,昆明 650204;2西藏高原生物研究所,拉薩 850001;3中國科學院研究生院,北京 100049

藏藥臭蚤草中 11個倍半萜化學成分研究

黃圣卓1,3,蔣思萍2,朱華結1＊

1中國科學院昆明植物研究所植物化學與植物資源持續(xù)利用國家重點實驗室,昆明 650204;2西藏高原生物研究所,拉薩 850001;3中國科學院研究生院,北京 100049

首次進行中國特有菊科藏藥臭蚤草 (Pulicaria insignisDrumm.ex Dunn)的化學成分研究,從全草中分離得到 11個倍半萜類化合物,通過波譜數(shù)據(jù)分析分別鑒定為 Xanthanolide(1),(3αR,4αR,5R,7αS,9αS)-5-Hydroxy-5-methyl-3,8-dimethylenedecahydroazuleno[6,5-β]furan-2(3H)-one(2)10α-Hydroxy-14H-inuviscolide(3), (3R,3αR,4αS,7αS,8S,9αS)-3,4α,8-Tr imethyldecahydroazuleno[6,5-β]furan-2,5-dione(4)8-Epi-conferti(5), (3αR,5S,11αS,E)-5-Hydroxy-10-methyl-3,6-dimethylene-3α,4,5,6,7,8,11,11α-octahydrocyclodeca[α]furan-2 (3H)-one(6),(3αR,8αR,9αR)-5,8α-Dimethyl-3-methylene-3α,4,8,8α,9,9α-hexahydronaphtho[2,3-β]furan-2,6 (3H,7H)-dione(7),Pterodonoic acid(8),8-Epi-isovangustin(9),(3αR,4αS,8R,8αR,9αR)-8-Hydroxy-5,8α-dimethyl-3-methylene-3,3α,4,4α,8,8α,9,9α-octahydronaphtho[2,3-β]furan-2(7H)-one(10),(5S,8αR,9αR)-3-(Hydroxymethyl)-5,8α-dimethyl-6,7,8,8α,9,9α-hexahydronaphtho[2,3-β]furan-2(5H)-one(11)。所有化合物均首次從該植物中分離得到,同時補充了文獻中化合物 6和 9所缺少的碳譜數(shù)據(jù)。

藏藥;臭蚤草;倍半萜

臭蚤草Pulicaria insignisDrumm.ex Dunn是菊科 Compositea(Asteraceae)旋復花族Inluneae蚤草屬Pulicaria的多年生草本。僅產西藏南部江孜、拉薩、察日壩、沙伽、尼木、日喀則、南太林等處。生于山脊巖石上,常見于石礫坡地和草叢中,海拔 4000～4310 m,根狀莖粗壯、多分枝,“成片叢生,植物有粘液,具惡臭”[1],為西藏特有的藥用植物。臭蚤草的花在西藏供藥用,藏名“Ming·chen·serpo”;全草味苦性涼,具有鎮(zhèn)咳舒肝,清血熱,透骨蒸。用于肺癆咳嗽,兩肋疼痛,勞熱骨蒸,花序有清熱、止痛的功效,還有驅跳蚤的作用[2]。目前人們從該屬植物中分離到的化合物主要為倍半萜、二萜和黃酮等化合物[3-10],未見臭蚤草的化學成分報道。為了了解臭蚤草的活性成分,我們對該植物全草進行化學成分研究,從中分離得到 11化合物,所有化合物均首次從該植物中分離得到,同時補充了文獻中化合物6和 9所缺少的碳譜數(shù)據(jù)。

1 儀器和材料

VGA Autospec-3000型質譜儀;Bruker AM-400和DRX-500超導核磁共振儀,T MS(四甲基硅烷)為內標;JASCO D IP-370和 OA AA-55型數(shù)字旋光儀,單位為 c單位為 g/mL;薄層色譜和正相柱色譜硅膠(青島海洋化工);Sephadex LH-20(Phar macia公司);ODS(Y MC公司);MCI樹脂(三菱公司)。

植物樣品于 2007年 10月采于西藏拉薩,植物標本由西藏高原生物所蔣思萍研究員鑒定為Pulicaria insignisDrumm.exDunn,標本保存于西藏高原生物所標本室。

2 提取和分離

90 Kg干燥臭蚤草全草,粉碎后用甲醇提取濃縮獲得浸膏,石油醚脫脂后用乙酸乙酯萃取 10次濃縮獲得浸膏 780 g。乙酸乙酯部石油醚 /乙酸乙酯(20∶1-0∶1)分為 5個部分 (20∶1,10∶1,5∶1,2∶1, 0∶1),其中石油醚/乙酸乙酯 (10∶1)部分經MCI樹脂脫色(收集甲醇洗脫部分),再經石油醚/丙酮 (50∶1-10∶1)和氯仿/丙酮 (100∶1-20∶1)多次反復梯度洗脫,經ODS柱(甲醇/水 1∶4-4∶1)梯度洗脫后,再分別經 SephadexLH-20凝膠(氯仿/甲醇 1∶1洗脫)純化,分別獲得化合物 1(256 mg),2(11.1 mg),3(851 mg),4(16.2 mg),5(382 mg),6(8.8 mg),7(20 mg),8 (8.8 mg),9(171 mg),10(4 mg),11(18.8 mg)。

3 結構鑒定

Xanthanolide(1) 白色塊狀晶體,C15H20O3; mp.128～129℃,+26.6(c0.0064 g/mL,下同,MeOH);1H NMR(CDCl3,500 MHz)δ(ppm): 6.12(d,J=3.2 Hz,1H),5.43(d,J=3.2 Hz,1 H),5.33(m,1H),4.54(td,J=8.7,2.7 Hz,1H), 3.22(m,1H),2.39(m,1H),2.30(m,3H),2.10 (m,3 H),2.08(s,3H),1.87(m,1H),1.80(dd, 1H),1.02(d,J=7.0 Hz,3H),ESI-MS(positive)m/z:249[M+1]+,271[M+Na]+,228;碳譜數(shù)據(jù)見表 1。其波譜數(shù)據(jù)與文獻[11]報道一致。

(3αR,4αR,5R,7αS,9αS)-5-Hydroxy-5-methyl-3,8-d imethylenedecahydroazuleno[6,5-β]furan-2(3H)-one(2) 白色塊狀晶體,C15H20O3;mp.132～133℃;+62.3(c0.0014,CHCl3);1H NMR (CDCl3,400 MHz)δ(ppm):2.69(1H,d,J=10.5 Hz),1.96(1H,m),1.65(1H,m),1.85(m,1 H), 1.65(m,1H),1.71(1H,m),2.28(1H,t,J= 13.3,3.7 Hz),1.24(1H,m),2.66(1H,m),4.31 (1H,ddd,J=10.6,9.3,1.6 Hz),2.55(1H,dd,J= 15.5,10.6 Hz),6.22(1H,d,J=3.5 Hz),5.54 (1H,d,J=3.5 Hz),5.09(1H,s),4.97(1H,s), 1.20(3H,s),EI-MSm/z:248[M]+,230[MH2O]+,215,190,145;碳譜數(shù)據(jù)見表 1。其波譜數(shù)據(jù)與文獻[12]報道一致。

10α-Hydroxy-14H-inuviscolide(3) 無色油狀物質,C15H22O4;+56.7(c0.00335 g/mL, CHCl3);1H NMR(CDCl3,500 MHz)δ(ppm):6.00 (1H,d,J=3.5 Hz),5.40(1H,d,J=3.5 Hz),4.25 (1H,dd,J=12.5,10.0 Hz),2.43(1H,ddd,J= 11.0,10.5,3.5 Hz),2.33(1H,d,J=14.5 Hz), 2.20(1H,ddd,J=12.5,4.0,2.0 Hz),1.89(1H, dd,J=14.5,12.0 Hz),1.83(1H,m),1.67(1H, m),1.54(2H,m),1.48(1H,ddd,J=11.5,11.0, 2.0 Hz),1.67(3H,s),1.05(3H,s),EI-MSm/z: 267[M+1]+,230[M-H2O]+,191,190,95,81;碳譜數(shù)據(jù)見表 1。其波譜數(shù)據(jù)與文獻[12]報道一致。

(3R,3αR,4αS,7αS,8S,9αS)-3,4α,8-Tr imethyldecahydroazuleno[6,5-β]furan-2,5-dione(4)無色油狀物質,C15H22O3;+59.2(c0.0071 g/mL,CHCl3),1H-NMR (CDCl3,400 MHz)δ (ppm):1.81(1H,m),2.01(1H,m),2.05(1H, m),2.20(1H,ddd,J=19.0,9.0,9.0 Hz),2.49 (1H,ddd,J=19.0,9.0,2.0 Hz),2.57(1H,dd,J= 15.0,5.0 Hz),1.30(1H,dd,J=15.0,11.5 Hz), 1.85(1H,ddd,J=11.5,11.0,9.0 Hz),4.29(1H, ddd,J=12.0,9.0,3.0 Hz),2.29(1H,m),1.74 (1H,ddd,J=12.0,12.0,6.0 Hz),2.31(1H,m), 2.33(1H,m),1.22(3H,d,J=7.0 Hz),1.18(3H, d,J=7.0 Hz),1.08(3H,s),FAB-MS(positive)m/ z:251[M+1]+,233,111;碳譜數(shù)據(jù)見表 1。其波譜數(shù)據(jù)與文獻[13]報道一致。

8-Epi-confertin(5) 白色塊狀晶體,C15H20O3;mp.125～126℃,+163.5(c0.00315 g/mL,CHCl3),1H NMR(CDCl3,400 MHz)δ(ppm): 6.18(1H,d,J=3.5 Hz),5.53(1H,d,J=3.5 Hz), 4.28(1H,td,J=8.8,2.4 Hz),2.73(1H,m),2.67 (1H,dd,J=14.8,5.3 Hz),2.50(1H,m),2.30 (2H,m)2.17(1H,dd,J=19.5,9.7 Hz),2.15 (1H,m),2.08(1H,m),1.85(2H,m),1.43(1H, t,J=11.8 Hz),1.16(3H,d,J=7.6 Hz,),1.07 (3H,s),EI-MSm/z:248[M]+,228,230,215,204, 189,136,81;碳譜數(shù)據(jù)見表 1。其波譜數(shù)據(jù)與文獻[12]報道一致。

(3αR,5S,11αS,E)-5-Hydroxy-10-methyl-3,6-d imethylene-3α,4,5,6,7,8,11,11α-octahydrocyclodeca[β]furan-2(3H)-one(6) 淡黃色油狀物質,C15H20O3;-52.63 (c0.00152 g/mL, MeOH),1H NMR(CDCl3,400 MHz)δ(ppm):6.30 (1H,d,J=3.0 Hz),6.18(1H,d,J=3.0 Hz),5.50 (1H,dd,J=7.7,7.5 Hz),5.33(1H,s),5.16(1H, s),4.14(1H,m),4.08(1H,d,J=6.3 Hz),2.75 (2H,m),2.38(2H,m),2.04(1H,t,J=11.6 Hz), 1.96(1H,dd,J=7.0,14.5 Hz),1.85(1H,m), 1.73(2H,m),1.70(3H,s),ESI-MS(positive)m/ z:249[M+1]+,271[M+Na]+;碳譜數(shù)據(jù)見表 1。其波譜數(shù)據(jù)與文獻[14]報道一致。

(3αR,8αR,9αR)-5,8α-D imethyl-3-methylene-3α,4,8,8α,9,9α-hexahydronaphtho[2,3-β]furan-2,6(3H,7H)-dione(7) 白色粉末,C15H18O3;+17.3(c0.00075 g/mL,MeOH),1H NMR (CDCl3,500 MHz)δ(ppm):6.38(1H,d,J=3.2 Hz),5.72(1H,d,J=2.6 Hz),4.61(1H,ddd,J= 11.5,8.5,4.7 Hz),3.22(1H,m),3.04(1H,dd,J=13.1,7.3 Hz),2.60(1H,td,J=14.1,5.5 Hz), 2.46(1H,dt,J=14.1,3.1 Hz),2.23(1H,t,J= 14.5 Hz),2.05(1H,m),1.94(1H,dd,J=13.8, 4.7 Hz),1.83(3H,s),1.60-1.79(2H,m),1.24 (3H,s),FAB-MS(positive)m/z:247[M+1],154, 136,107,83;碳譜數(shù)據(jù)見表 1。其波譜數(shù)據(jù)與文獻[15]報道一致。

Pterodonoic acid(8) 白色粉末,C15H20O3;+17.3(c0.00075 g/mL,CH3OH)1H NMR (CDCl3,500 MHz)δ (ppm):6.41(1H,s),5.76 (1H,s),2.88(1H,d,J=14.5 Hz),2.41-2.84 (3H,m),2.06(1H,t,J=13.6 Hz),1.70-1.85 (5H,m),1.79(s,3H),1.51(2H,dt,J=4.1,12.7 Hz),1.25(3 H,s),ESI-MS(positive)m/z:249[M +1]+,271[M+K]+,519[2 M+Na]+;碳譜數(shù)據(jù)見表 1。其波譜數(shù)據(jù)與文獻[16]報道一致。

8-Epi-isovangustin(9) 白色塊狀晶體,C15H24O3;mp.138～140℃,+17.3(c0.0035 g/ mL,CH3OH),1H NMR(CDCl3,400MHz)δ(ppm): 6.13(1H,d,J=1.1 Hz),5.63(1H,d,J=1.1 Hz), 2.46(1H,tt,J=8.5,3.5 Hz),1.94(1H,d,J=12.4 Hz),1.70(1H,dd,J=12.4 Hz),1.70～1.15 (11H,m),1.07(3H,s),0.93(3H,s),ESI-MS (positive)m/z:251[M+Na]+,503[2M+1]+;碳譜數(shù)據(jù)見表 1。其波譜數(shù)據(jù)與文獻[17]報道一致。

(3αR,4αS,8R,8αR,9αR)-8-Hydroxy-5,8αd imethyl-3-methylene-3,3α,4,4α,8,8α,9,9α-octahydronaphtho[2,3-β]furan-2(7H)-one(10) 無色油狀物質,C15H20O3;[α]21D-31.5(c0.0016 g/mL, MeOH),1H NMR(CDCl3,400 MHz)δ(ppm):3.7 (1H,dd,J=10.0,7.0 Hz),4.05(1H,ddd,J= 12.5,12.5,4.0 Hz),5.34(1H,s),6.09(1H,d,J= 3.0 Hz),5.45(1H,d,J=3 Hz),2.51(1H,dd,J= 12.5,4.0 Hz),2.30-2.40(2H,m),2.25(1H,ddd,J=12.5,12.5,3.0 Hz),2.17(1H,d,J=3.0 Hz), 2.01(1H,dddq,J=12.0,10.0,2.15,2.15 Hz), 1.69(3H,s),1.42(2H,m),0.89(3H,s),ESI-MS (positive)m/z:249[M+1]+,271[M+Na]+,519 [2M+Na]+,131,185,157,119,105;碳譜數(shù)據(jù)見表1。其波譜數(shù)據(jù)與文獻報道一致[18].

(5S,8αR,9αR)-3-(Hydroxymethyl)-5,8αd imethyl-6,7,8,8α,9,9α-hexahydronaphtho[2,3-β]furan-2(5H)-one(11) 淡黃色油狀物質,C15H20O3;+249.2(c0.00915 g/mL,CHCl3);1H NMR(CDCl3,400 MHz)δ(ppm):1.60(1H,m), 1.68(1H,dd,J=13.0,4.0 Hz),1.56(1H,m), 1.95(1H,m),1.56(1H,m),1.75(1H,m),2.75 (1H,ddq,J=7.5,6.5,2.0 Hz),6.36(1H,s),4.78 (1H,dd,J=13.0,6.0 Hz),2.14(1H,dd,J=13.0, 6.0 Hz),1.54(1H,dd,J=13.0,13.0 Hz),4.40 (2H,s),1.32(3H,s),1.29(3H,d,J=7.5 Hz), 2.68(br),EI-MSm/z:248[M]+,230[M-H2O]+, 215,159,146;HR-ES IMS(positive)[M+Na]+271.1314,Calcd for[C15H20O3+Na]+m/z: 271.1310;碳譜數(shù)據(jù)見表 1。其波譜數(shù)據(jù)與文獻報道一致[19].

圖 1 臭蚤草中分離到的化合物 1～11Fig.1 Compounds 1-11 isolated fromPulicaria insignis

表 1 化合物 1～11的13C NMR數(shù)據(jù) (13C,100 MHz in CDCl3)δ(ppm)Table 113C NMR of compounds 1-11(13C,100 MHz in CDCl3)δ(ppm).

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Sesquiterpenoids from Tibetan Folk DrugPulicaria insignis

HUANG Sheng-zhuo1,3,J IANG Si-ping2,ZHU Hua-jie1＊

1State key laboratory of Phytochem istry and Plant Rescources of W est of China,Kunm ing Institute of Botany,Academy of Sciences Kunm ing 650204,Yunnan China;2Plateau Institute of B iology Lhasa 850001,Tibet China;3Graduate University of Chinese Academy of Sciences Beijing 100049,China

Eleven known sesquiterpenoids were isolated from the Tibetan Folk DrugPulicaria insignis.On the basis of MS and NMR data,theywere identified as Xanthanolide(1),(3αR,4αR,5R,7αS,9αS)-5-Hydroxy-5-methyl-3,8-d imethylenedecahydroazuleno[6,5-β]furan-2(3H)-one(2),10α-hydroxy-14H-inuviscolide(3),(3R,3αR,4αS,7αS,8S,9αS)-3,4α,8-Trimethyldecahydroazuleno[6,5-β]furan-2,5-dione(4),8-Epi-conferti(5),(3αR,5S,11αS,E)-5-Hydroxy-10-methyl-3,6-dimethylene-3α,4,5,6,7,8,11,11α,-octahydrocyclodeca[α]furan-2(3H)-one(6),(3αR,8αR,9αR)-5,8α-Dimethyl-3-methylene-3α,4,8,8α,9,9α-hexahydronaphtho[2,3-β]furan-2,6(3H,7H)-dione(7), Pterodonoic acid(8),8-Epi-isovangustin(9),(3αR,4αS,8R,8αR,9αR)-8-Hydroxy-5,8α-d imethyl-3-methylene-3, 3α,4,4α,8,8α,9,9α-octahydronaphtho[2,3-β]furan-2(7H)-one(10)(5S,8αR,9αR)-3-(Hydroxymethyl)-5,8αd imethyl-6,7,8,8α,9,9α-hexahydronaphtho[2,3-β]furan-2(5H)one(11).All compounds were isolated from this plant for the first t ime,and the13C NMR data of compounds 6 and 9 were shawed in this paper.

Tibetan drug;Pulicaria insignis;chemical constituents;sesquiterpenoids

R284.2;Q946

A

1001-6880(2010)05-0736-05

2009-04-30 接受日期:2009-10-20

國家自然科學基金(NSFC#30770235);中國科學院院地合作基金(YZ-06-1)

＊通訊作者 Tel:86-871-5216179;E-mail:hjzhu@mail.kib.ac.cn