鸡肠狼毒植物根中二化学成分及其药理活性研究

鸡肠狼毒植物根中二化学成分及其药理活性研究

鸡肠狼毒属于甲状腺齐属植物，主要分布在中国云南，是一种多年生草本植物。鸡肠狼毒以根入药,具有利水消肿、泻下通便的功效,主治各种水肿、便秘、外伤出血、疮及无名肿毒等。吴大刚、陈道峰等曾对其进行植化研究。我们对该植物的石油醚萃取部分进行了较详细的研究,从中分离得到8个已知的曼西醇型二萜(图1)。1材料和方法1.1溶剂、带标植物的研究核磁共振谱用VarianUnityNOVA400/54核磁共振仪(溶剂为CDCl3,TMS为内标)。鸡肠狼毒植物(2010年采集自云南)由云南大学蔡乐博士鉴定;层析用硅胶为青岛海洋化工厂出品;TLC用溶剂系统:S1氯仿-甲醇,S2石油醚-乙酸乙酯,S3甲醇-水,层析所用溶剂为分析纯。1.2粗树莓原汁的制备鸡肠狼毒根粉2.5kg,以约15L乙醇反复提取得到乙醇提取物,减压蒸馏得到浸膏,总浸膏用水分散后,依次用石油醚、乙酸乙酯、正丁醇萃取。石油醚萃取物(23.5g)经硅胶柱层析(100～200目,500g粗硅胶),石油醚-乙酸乙酯(10∶1～1∶1)梯度洗脱,洗脱液合并,减压浓缩得到A(1.6g)、B(2.8g)、C(4.4g)、D(9.7g)、E(2.5g)五部分。A部分经反复硅胶柱层析得77mg化合物7,B部分经反复硅胶柱层析得25mg化合物2和19mg化合物5,C部分经反复硅胶柱层析得到12mg化合物6,D部分经反复硅胶柱层析得到38mg化合物1和23mg化合物3,E部分经反复硅胶柱层析得到6mg化合物4和15mg化合物8。2结果2.1h,s,d,d℃,HR-ESI-MS显示分子式为C42H54O15(m/z821.3390[M+Na]+,calc.for821.3355)。1H-NMR(400MHz,CDCl3)δH6.19(1H,ddd,J=10.4,6.4,1.2Hz,H-8),5.94(1H,dd,J=10.8,1.2Hz,H-5),5.91(1H,dd,J=10.0,5.2Hz,H-9),5.80(1H,s,H-14),5.44(1H,d,J=4.0Hz,H-3),4.84(1H,d,J=6.4Hz,H-7),4.12(1H,d,J=8.8Hz,H-17α),3.72(1H,dd,J=10.8,4.0Hz,H-4),3.51(1H,dd,J=8.8,1.2Hz,H-17β)。13C-NMR(100MHz,CDCl3)47.5(1),86.4(2),77.6(3),47.7(4),68.5(5),53.4(6),62.8(7),125.8(8),129.8(9),85.8(10),44.6(11),37.0(12),90.0(13),73.5(14),90.0(15),18.9(16),69.8(17),25.1(18),21.2(19),24.1(20),173.3(3-OPr),27.9(2′),8.7(3′),169.1(5-OAc),20.7(2′),170.6(7-OAc),22.4(2′),170.3(10-OAc),20.8(2′),168.4(15-OAc),22.2(2′),165.6(14-OBz),130.1(1′),130.0(2′,6′),128.3(3′,5′),133.2(4′),173.5(2-OMp),34.3(2′),18.2(3′),18.8(4′)。以上NMR数据与文献值基本一致。2.2c-nmr,ccl3,5-opr,5-opr,5-opr,5-opr,5-opr,5-opr,5-opr,5-1,3,5-,3,5-opr,5-1,3,5-,3,5-opr,5-1,3,5-1,3,4.3,5-,7.3,5-opr,5-1,3,5-opr,5-1,3,5-,7.3,5-1,3,5-opr,5-1,3,5-opr,5-1,3,5-opr,5-1,3,4.3,5-,4.3,5-opr,5-,7-1,3,5-opr,5-1,3,4.3,5-,4.3,4.3,4.3,4.3,4.3,4.3,5-opr,5-1,3,5-opr,5-1,3,5-,5-1,3,5-1,3,5-1,3,5-1,3,5-1,3,5-1,3,5-1,3,5-1,3,5-,5-1,3,5-3,5-3,5-1,3,5-3,5-1,3,5-3,5-3,5-3,5-3,5-3,5-3,5-3,5-3,5-3,5-3,5-3,5-3,5-3,5-3,5-3,5-3,5-3,5-3,4.3,4.2,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4H-NMR(400MHz,CDCl3)δH6.19(1H,ddd,J=10.4,6.8,1.2Hz,H-8),6.02(1H,dd,J=10.8,1.2Hz,H-5),5.91(1H,dd,J=10.4,5.8Hz,H-9),5.83(1H,s,H-14),5.51(1H,d,J=3.6Hz,H-3),4.87(1H,d,J=6.8Hz,H-7),4.19(1H,d,J=8.8Hz,H-17α),3.95(1H,dd,J=10.8,3.6Hz,H-4),3.53(1H,dd,J=8.8,1.2Hz,H-17β)。13C-NMR(100MHz,CDCl3)46.6(1),87.6(2),78.6(3),47.8(4),68.7(5),53.6(6),63.0(7),125.9(8),129.9(9),85.8(10),44.7(11),37.1(12),90.1(13),73.3(14),90.2(15),18.3(16),69.9(17),25.2(18),21.0(19),24.3(20),173.6(3-OPr),28.1(2′),8.8(3′),169.3(5-OAc),21.0(2′),170.4(7-OAc),21.3(2′),170.7(10-OAc),22.5(2′),168.5(15-OAc),22.4(2′),164.8(2-OBz),129.2(1′),129.8(2′,6′),128.2(3′,5′),132.8(4′),165.9(14-OBz),128.4(1′),129.5(2′,6′),128.1(3′,5′),132.6(4′)。以上NMR数据与文献值基本一致。2.3d,l,nmr,5-opr,5-opr,5--1,3.2.3,4.4h,4.4h,5-7,4.4h,5-7,4.4h,3.4h,3.4h,3.4h,3.4h,3.4h,3.4h,3.4h,3.4h,3.3,3.4h,3.4h,3.3,5-opr,7.2,3.3,5-opr,5-opr,5-opr,5-opr,7.2,3.2,3.2,3.2,3.2,3.2,3.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,5-opr,5-,5-opr,5-1,3,5-oc,5-10-,5-oc,5-1,3,4.3,4.3,4.3,4.4,5-10.3,4.3,4.3,4.3,4.3,4.3,4.2,4.3,4.3,4.3,4.3,4.3,4.3,4.2,4.2,4.3,5-,5-1,3.2,4.2,4.3,4.4h,4.4h,4.4h,4.4h,4.4h,4.4h,4.4h,5.4h,4.4h,4.4h,4.4h,4.4h,4.4h,4.4h,4.4h,5.4h,4.4h,4.4h,4.4h,13c-7,4.4h,5-7,10.H-NMR(400MHz,CDCl3)δH6.17(1H,ddd,J=10.0,6.4,1.2Hz,H-8),5.96(1H,dd,J=11.2,1.2Hz,H-5),5.90(1H,dd,J=10.0,5.6Hz,H-9),5.93(1H,s,H-14),5.36(1H,t,J=3.2Hz,H-3),4.86(1H,d,J=6.4Hz,H-7),4.12(1H,d,J=8.4Hz,H-17α),3.50(1H,dd,J=8.4,1.2Hz,H-17β),3.48(1H,dd,J=11.2,3.2Hz,H-4)。13C-NMR(100MHz,CDCl3)45.6(1),38.1(2),76.8(3),50.0(4),68.9(5),53.7(6),63.3(7),125.7(8),129.9(9),85.9(10),44.9(11),37.3(12),90.0(13),73.4(14),89.2(15),14.0(16),70.0(17),25.4(18),21.1(19),24.9(20),174.1(3-OPr),28.2(2′),9.1(3′),169.2(5-OAc),21.1(2′),170.7(7-OAc),22.7(2′),170.4(10-OAc),21.0(2′),168.6(15-OAc),22.5(2′),165.5(14-OBz),130.0(1′),130.0(2′,6′),128.6(3′,5′),133.4(4′)。以上NMR数据与文献值基本一致。2.4c-nmr,9.5,10.4h,7.4h,5-opr,7.4h-4,5-opr,7.4,4.4e,4.4e,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,5-opr,5-opr,5-opr,5-opr,5-opr,5-opr,5-opr,5-opr,5-opr,5-opr,5-d,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4-3,4.3,4-3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,4.3,13c-3,13c-3,13c-3,13c-3,5-1,3,4.3,4.3,4.3,H-NMR(400MHz,CDCl3)δH5.79(1H,dd,J=11.2,1.2Hz,H-5),5.56(1H,d,J=5.2Hz,H-8),5.39(1H,t,J=4.0Hz,H-3),5.20(1H,s,H-14),4.25(1H,d,J=9.6Hz,H-17α),3.69(1H,dd,J=9.6,1.2Hz,H-17β),2.76(1H,dd,J=11.2,4.0Hz,H-4)。13C-NMR(100MHz,CDCl3)50.6(1),30.8(2),78.9(3),50.6(4),69.3(5),63.0(6),205.4(7),72.4(8),35.5(9),78.4(10),41.4(11),41.9(12),89.3(13),83.1(14),81.3(15),14.5(16),67.2(17),34.7(18),25.4(19),22.3(20),173.1(3-OPr),26.3(2′),8.7(3′),170.2(5-OAc),22.6(2′),169.3(10-OAc),22.0(2′),160.1(8-OBz),130.3(1′),130.1(2′,6′),128.5(3′,5′),133.4(4′),167.0(14-OBz),130.3(1′),129.9(2′,6′),128.4(3′,5′),133.4(4′)。以上NMR数据与文献值基本一致。2.5d,h,5.H-NMR(400MHz,CDCl3)δH6.19(1H,dd,J=10.4,6.4Hz,H-8),5.95(1H,d,J=10.8Hz,H-5),5.91(1H,dd,J=10.0,5.6Hz,H-9),5.82(1H,s,H-14),5.41(1H,d,J=4.0Hz,H-3),4.85(1H,d,J=6.4Hz,H-7),4.16(1H,d,J=8.4Hz,H-17α),3.74(1H,dd,J=11.2,4.0Hz,H-4),3.52(1H,dd,J=8.4,0.8Hz,H-17β)。以上NMR数据与文献值基本一致。2.6j,2,3,5.4h,t,h-17,dH-NMR(400MHz,CDCl3)δH5.78(1H,dd,J=11.2,1.2Hz,H-5),5.54(1H,d,J=5.6Hz,H-8),5.36(1H,t,J=4.0Hz,H-3),4.96(1H,s,H-14),4.92(1H,d,J=12.0Hz,H-12),4.21(1H,d,J=9.6Hz,H-17α),3.65(1H,dd,J=9.6,1.2Hz,H-17β),2.73(1H,dd,J=11.2,4.0Hz,H-4)。以上NMR数据与文献值基本一致。2.7j,2h,d,n,nh-1h,d,d,nh-17,d,d,n,h-17,2.2h,d,d,n,n,n,n,n,n,n,n,n,n,h-17H-NMR(400MHz,CDCl3)δH6.09(1H,dd,J=11.2,1.2Hz,H-5),5.46(1H,t,J=4.0Hz,H-3),5.18(1H,d,J=7.2Hz,H-8),5.04(1H,s,H-14),4.31(1H,d,J=10.0Hz,H-17α),4.06(1H,d,J=12.8Hz,H-12),3.65(1H,dd,J=10.0,1.2Hz,H-17β),3.03(1H,dd,J=11.2,4.0Hz,H-4)。以上NMR数据与文献值基本一致。2.81高年设计组集体规划2.H-NMR(400MHz,CDCl3)δH6.29(1H,ddd,J=10.0,6.4,1.2Hz,H-8),5.97(1H,dd,J=11.2,