扁枝石松中石杉型三萜成分研究

1、扁枝石松中石杉型三萜成分研究【摘要】目的對(duì)扁枝石松全草80%乙醇提取物中的石杉型三萜類化學(xué)成分進(jìn)展研究。方法利用正相和反相硅膠柱色譜方法進(jìn)展別離純化，通過S,1H-NR,13-NR波譜數(shù)據(jù)鑒定化合物構(gòu)造。結(jié)果從扁枝石松共別離得到7個(gè)石杉型三萜類化合物，分別鑒定為serrat-14-en-3,21-dil；serrat-14-en-3,21-dil；3-hydrxy-21-aetxy-14-serraten；3,21-dihydrxy-14-serraten-24-iaid(lyernuiaidA,；14-serraten-3,21,24-tril(lylavanl,3,20,21-trihyd

2、rxy-14-serraten-24-iaid；21-episerratenedil-3-L-arabinpyranside(inundsideE,。結(jié)論化合物7為首次從該屬植物中別離得到，化合物3為首次從該植物中分到?！娟P(guān)鍵詞】扁枝石松；化學(xué)成分；石杉型三萜Abstrat：bjetiveTstudytheserratene-typetriterpenidsfrthe80%alhlextratfDiphasiastruplanatu.ethdsThepundsereislatedandpurifiedbysiliagel,18reverse-phasesiliagellunhratgraphi

3、ethds,andthEirstruturesereeluidatedbyspetralanalysis.ResultsSevenserratene-typetriterpenidserebtainedandidentifiedasserrat-14-en-3,21-dil(1),serrat-14-en-3,21-dil(2),3-hydrxy-21-aetxy-14-serraten(3),3,21-dihydrxy-14-serraten-24-iaid(lyernuiaidA,4),14-serraten-3,21,24-tril(lylavanl,5),3,20,21-trihydr

4、xy-14-serraten-24-iaid(6),21-episerratenedil-3-L-arabinpyranside(inundsideE,7).nlusinpund7asislatedfrDiphasiastruandpund3frtheplantfrthefirsttie.Keyrds：Diphasiastruplanatu;heialnstituents;Serratene-typetriterpenids扁枝石松DiphasiastruplanatuL.屬石松科扁枝石松屬植物，又名伸筋草、扁心草，具舒筋活血、祛風(fēng)散寒、通經(jīng)、消炎等作用，主治風(fēng)濕骨痛，月經(jīng)不調(diào)，跌打損傷，燒、

5、燙傷等1。石松科植物化學(xué)成分主要含有石松生物堿2和石杉型三萜。石杉型三萜是一類特殊的五環(huán)三萜，其中環(huán)為七元環(huán)，另外四個(gè)環(huán)為六元環(huán)，七個(gè)角甲基或其氧化形式，14-15位常有雙鍵，3位和21位為氧接官能團(tuán)。據(jù)報(bào)道該類化合物具有很強(qiáng)的抗腫瘤促進(jìn)活性、腫瘤化學(xué)預(yù)防以及抑制白色念珠菌分泌型天冬氨酸蛋白酶活性35。因此我們對(duì)在貴州民間被廣泛用于治療風(fēng)濕性關(guān)節(jié)炎、跌打損傷等疾病的扁枝石松進(jìn)展三萜類化學(xué)成分的系統(tǒng)研究，從中別離得到7個(gè)石杉型三萜類化合物，化合物7為首次從該屬植物中分得，化合物3為首次從該植物中分到。2方法與結(jié)果2.1提取別離15kg扁枝石松枯燥全草，粉碎，用80%乙醇料液比為15回流浸提4次，

6、提取時(shí)間分別為3h，3h，2h，2h，合并4次濾液，減壓回收乙醇，得粗浸膏。浸膏加適量水混懸，依次以石油醚、氯仿、醋酸乙酯和正丁醇萃取，分別得石油醚、氯仿、醋酸乙酯和正丁醇萃取物210，150，380g和410g。石油醚萃取局部經(jīng)硅膠柱層析，用石油醚-丙酮(10110)梯度洗脫，經(jīng)TL檢識(shí)，合并得5局部。其中Fr3局部經(jīng)硅膠柱層析，用石油醚-丙酮(81)洗脫，得到化合物1(200g)和2(120g)；Fr4局部經(jīng)硅膠柱層析，用石油醚-醋酸乙酯(101)洗脫，得到化合物3(300g)。氯仿萃取局部經(jīng)硅膠柱層析，氯仿-甲醇(20111)梯度洗脫，經(jīng)TL檢識(shí)后合并得4局部，其中Fr3局部經(jīng)硅膠柱層析

7、，石油醚-丙酮(8101)梯度洗脫，經(jīng)TL檢識(shí)后合并得Fr3-1，F(xiàn)r3-2和Fr3-3三局部。Fr3-1經(jīng)硅膠柱層析，氯仿-甲醇(201)洗脫得化合物4(120g)；Fr3-2局部經(jīng)硅膠柱層析，氯仿-甲醇(151)洗脫得化合物5(70g)；Fr3-3局部經(jīng)反相硅膠柱層析，30%、50%、70%、90%甲醇-水洗脫，經(jīng)TL檢識(shí)后合并得Fr3-3-1和Fr3-3-2兩局部，經(jīng)反復(fù)硅膠柱層析，氯仿-甲醇、石油醚-醋酸乙酯梯度洗脫后得白色粉末6(60g)和7(40g)。2.2構(gòu)造鑒定2.2.1化合物1白色粉末，EI-S/z442+,427,409,391,234,220,207,189。1H-NR(

8、400Hz,Dl3-eD)：5.33(1H,brs,H-15),3.22(1H,dd,J=10.4,5.6Hz,H-21),3.17(1H,dd,J=10.4,6.4Hz,H-3),0.96,0.95,0.83,0.82,0.80,0.76,0.67(s,7H3)；13NR(100Hz,Dl3-eD)：38.3(-1),24.9(-2),78.1(-3),37.7(-4),55.4(-5),18.5(-6),44.8(-7),38.5(-8),56.8(-9),36.7(-10),26.6(-11),27.0(-12),62.5(-13),137.8(-14),121.8(-15),23.5(

9、-16),49.2(-17),38.4(-18),35.8(-19),26.8(-20),78.4(-21),36.8(-22),27.5(-23),15.0(-24),15.2(-25),19.3(-26),55.7(-27),12.9(-28),14.1(-29),26.8(-30)。其光譜數(shù)據(jù)與文獻(xiàn)6報(bào)道的serrat-14-en-3,21-dil根本一致。2.2.2化合物2白色粉末，EI-S/z442+,427,409,302,284,269,255,234,220,207,189。1H-NR(400Hz,Dl3)：5.32(1H,brs,H-15),3.45(1H,brs,H-21)

10、,3.19(1H,dd,J=11.6,4.8Hz,H-3),0.97,0.93,0.89,0.83,0.80,0.77,0.69(s,7H3)。13-NR(400Hz,Dl3)：38.5(-1),25.2(-2),78.8(-3),38.1(-4),55.7(-5),18.9(-6),45.1(-7),38.9(-8),56.8(-9),35.9(-10),25.4(-11),27.1(-12),62.8(-13),138.5(-14),122.0(-15),24.0(-16),43.3(-17),37.1(-18),31.2(-19),27.5(-20),76.2(-21),37.4(-22

11、),28.1(-23),15.4(-24),15.7(-25),19.8(-26),56.2(-27),13.3(-28),21.8(-29),27.7(-30)。以上數(shù)據(jù)與文獻(xiàn)6報(bào)道的serrat-14-en-3,21-dil一致。2.2.3化合物3白色晶體(氯仿-甲醇),EI-S/z484+,469,424,409,391,262,207,189。1H-NR(400Hz,Dl3)：5.32(1H,brs,H-15),4.50(1H,dd,J=11.2,4.4Hz,H-21),3.18(1H,dd,J=11.2,4.4Hz,H-3),2.06(3H,s,H3),0.97,0.90,0.84,

12、0.83,0.80,0.77,0.69(s,7H3)；13-NR(100Hz,Dl3)：38.6(-1),27.5(-2),78.8(-3),38.1(-4),55.7(-5),18.9(-6),45.1(-7),38.9(-8),62.8(-9),35.9(-10),25.3(-11),27.1(-12),57.0(-13),138.3(-14),121.9(-15),23.8(-16),49.6(-17),37.7(-18),36.7(-19),24.1(-20),81.1(-21),37.1(-22),28.1(-23),15.7(-24),15.4(-25),19.8(-26),56.

13、0(-27),13.4(-28),15.8(-29),27.5(-30),171.1(=),21.3(H3)。其光譜數(shù)據(jù)與文獻(xiàn)7報(bào)道的3-hydrxy-21-aetxy-14-serraten一致。2.2.4化合物4白色粉末，EI-S/z472+,454,439,421,332,270,237,220,203,187。1H-NR(400Hz,5D5N)：5.46(1H,brs,H-15),3.65(1H,brs,H-21),3.36(1H,dd,J=12.0,4.0Hz,H-3),1.69,1.13,1.02,0.92,0.85,0.81(s,6H3)；13-NR(100Hz,5D5N)：39

14、.7(-1),29.4(-2),78.2(-3),49.4(-4),57.0(-5),21.3(-6),45.6(-7),37.3(-8),62.5(-9),38.9(-10),26.7(-11),27.7(-12),57.3(-13),139.0(-14),122.9(-15),24.6(-16),43.8(-17),36.4(-18),31.8(-19),25.7(-20),75.3(-21),38.0(-22),24.8(-23),180.9(-24),14.3(-25),19.8(-26),56.8(-27),13.9(-28),22.2(-29),28.7(-30)。其譜學(xué)數(shù)據(jù)與文獻(xiàn)

15、5報(bào)道的3,21-dihydrxy-14-serraten-24-iaidlyernuiaidA根本一致。轉(zhuǎn)貼于論文聯(lián)盟.ll.2.2.5化合物5白色粉末，Ei-S/z458+,440,422,407,391,269,205,187,1H-NR(400Hz,5D5N)：5.44(1H,brs,H-15),4.44(1H,brs,H-3),4.10(1H,d,J=11.2Hz,H-24),3.86(1H,d,J=10.8Hz,H-24),3.66(1H,brs,H-21),1.59,1.15,0.93,0.89,0.83,0.80(s,6H3)。13-NR(100Hz,5D5N)：34.2(-1

16、),26.8(-2),70.0(-3),44.3(-4),50.3(-5),19.7(-6),46.0(-7),38.0(-8),63.1(-9),38.6(-10),25.5(-11),27.6(-12),57.4(-13),139.1(-14),122.7(-15),24.6(-16),43.8(-17),36.4(-18),31.9(-19),26.7(-20),75.3(-21),37.7(-22),23.7(-23),65.8(-24),16.6(-25),20.1(-26),56.8(-27),13.8(-28),22.2(-29),28.7(-30).其光譜數(shù)據(jù)與文獻(xiàn)8報(bào)道的14

17、-serraten-3,21,24-tril(lylavanl)根本一致。2.2.6化合物6白色粉末，EI-S/z488+,470,452,437,408,372,250,237,219,201,187。1H-NR(400Hz,5D5N)：5.45(1H,brs,H-15),4.69(1H,brs,H-3),4.37(1H,d,J=10.4,H-20),3.82(1H,brs,H-21),1.75,1.19,1.12,0.95,0.92,0.84(s,6H3)。13-NR(100Hz,5D5N)：34.8(-1),27.9(-2),70.6(-3),48.6(-4),49.6(-5),21.2

18、(-6),45.7(-7),37.6(-8),62.6(-9),39.4(-10),25.8(-11),27.6(-12),57.6(-13),139.0(-14),122.8(-15),24.4(-16),41.3(-17),37.8(-18),43.3(-19),66.5(-20),79.6(-21),38.9(-22),25.5(-23),180.7(-24),14.1(-25),19.9(-26),56.9(-27),14.7(-28),21.7(-29),28.8(-30)。以上數(shù)據(jù)與文獻(xiàn)8報(bào)道的3,20,21-trihydrxy-14-serraten-24-iaid一致。2.2.

19、7化合物7白色粉末，ESI-S/z573-H-,609+l-；EI-S/z488,470,455,439,277,259,205,187。1H-NR(400Hz,5D5N)：5.46(1H,brs,15-H),4.78(1H,d,J=6.8Hz,H-1),3.38(1H,dd,J=11.6,4.4Hz,H-21),1.28,1.15,0.97,0.93,0.80,0.79,0.77(s,7H3)；13-NR(100Hz,5D5N)：38.1(-1),26.6(-2),88.7(-3),39.8(-4),56.2(-5),19.1(-6),45.5(-7),38.0(-8),63.0(-9),3

20、8.9(-10),25.5(-11),27.5(-12),57.3(-13),139.0(-14),122.7(-15),24.6(-16),43.8(-17),36.4(-18),31.8(-19),27.0(-20),75.3(-21),37.4(-22),28.7(-23),16.1(-24),16.9(-25),20.1(-26),56.7(-27),13.8(-28),22.2(-29),28.3(-30),107.6(-1),73.0(-2),74.7(-3),69.6(-4),66.8(-5)。其光譜數(shù)據(jù)與文獻(xiàn)9報(bào)道的21-episerratenedil-3-L-arabinpy

21、ranside(inundsideE)一致。3討論石杉型三萜類化合物在氯仿、醋酸乙酯、甲醇、二甲亞砜以及水中的溶解性都比擬差，僅在吡啶中能充分溶解，因此給別離純化及體外活性挑選工作帶來(lái)很大的困難。鑒于石杉型三萜具有較強(qiáng)的細(xì)胞毒和抗腫瘤及抗真菌活性，為了進(jìn)一步提醒該類化合物的生物活性，在其構(gòu)造中引入糖基，從而進(jìn)步該類化合物的水溶性是非常必要的。近幾年來(lái)，未見有關(guān)石杉型三萜糖苷類化合物及其生物活性的研究報(bào)道，作者從扁枝石松中別離得到一個(gè)的石杉型三萜糖苷類化合物，并正在對(duì)其生物活性進(jìn)展試驗(yàn)。繼續(xù)深化的研究對(duì)石杉型三萜糖苷類化合物的發(fā)現(xiàn)及其生物活性的說明具有一定的科學(xué)意義。【參考文獻(xiàn)】1何順志,徐文芬.貴州中草藥資源研究.貴陽(yáng)：貴州科技出版社,2022：148.2a,XQ,GangDV.ThelypdiualkalidsJ.Nat.Prd.Rep,2022,21：752.3Tanak