全 文 :天然产物研究与开发 Nat Prod Res Dev 2015,27:562-566,584
文章编号:1001-6880(2015)4-0562-06
收稿日期:2014-11-11 接受日期:2015-01-26
基金项目:公益性行业(农业)科研专项(201303117);海南省科技
成果示范推广专项(CGSF2011003);海南省中药现代化专
项(ZY201408);海南省产学研一体化专项(CXY20130044)
* 通讯作者 Tel:86-898-66961869;E-mail:daihaofu@ itbb. org. cn
四瓣崖摩化学成分研究
米承能1,2,3,梅文莉2,3,左文健2,3,蔡彩虹2,3,王 昊2,3,李绍鹏1,戴好富2,3*
1海南大学园艺园林学院,海口 570228;2 中国热带农业科学院热带生物技术研究所
农业部热带作物生物学与遗传资源利用重点实验室;3 海南省黎药资源天然产物研究与利用重点实验室,海口 571101
摘 要:从楝科崖摩属植物四瓣崖摩的树枝中分离得到 14 个化合物,运用波谱学方法分别鉴定为:22ξ-
hydroxytirucalla-7,24-dien-3,23-dione(1)、dymacrin D(2)、泽屋萜(3)、α-菠菜甾醇(4)、8-hydroxy-6-methoxy-3-
pentylisocoumarin(5)、ichenxanthone(6)、10-oxo-isodauc-3-en-15-al(7)、( + )-ent-ficusol(8)、3-甲氧基-4-羟基苯甲
酸甲酯(9)、香草醛(10)、2,4-二羟基-3,6-二甲基苯甲酸甲酯(11)、2-羟基-4-甲氧基-6-丙基苯甲酸甲酯(12)、2,
4-二羟基-6-甲基苯甲酸甲酯(13)和琥珀酸甲酯(14),以上化合物均为首次从崖摩属植物中分离得到。采用 El-
man比色法对全部化合物进行活性测试,结果表明化合物 11 对乙酰胆碱酯酶具有抑制活性。
关键词:四瓣崖摩;化学成分;乙酰胆碱酯酶
中图分类号:R284. 2 文献标识码:A DOI:10. 16333 / j. 1001-6880. 2015. 04. 002
Chemical Constituents from Amoora tetrapetala
MI Cheng-neng1,2,3,MEI Wen-li2,3,ZUO Wen-jian2,3,CAI Cai-hong2,3,WANG Hao2,3,LI Shao-peng1,DAI Hao-fu2,3*
1Hainan University,College of Horticulture,Haikou 570228,China;2Key Laboratory of Biology and Genetic Resources of
Tropical Crops,Ministry of Agriculture,Institute of Tropical Bioscience and Biotechnology,Chinese Academy of Tropical Agricultural
Sciences;3Hainan Key laboratory for research and development of natural products from Li folk medicine,Haikou 571101,China
Abstract:Fourteen compounds were isolated from the branches of Amoora tetrapetala and structurally identified by spec-
tral analysis. The compounds were identified as 22ξ-hydroxytirucalla-7,24-dien-3,23-dione (1),dymacrin D (2),zeorin
(3),α-spinasterol (4),8-hydroxy-6-methoxy-3-pentylisocoumarin (5),lichenxanthone (6),10-oxo-isodauc-3-en-15-al
(7),( + )-ent-ficusol (8),3-methoxy-4-hydroxybenzoate (9),vanillin (10),methyl 2,4-hydroxy-3,6-dimethylbenzo-
ate (11),methyl 2-hydroxy-4-methoxy-6-propylbenzoate (12),methyl 2,4-dihydroxy-6-methylbenzoate (13) and meth-
yl hydrogen succinate (14) . All the compounds were isolated from this genus for the first time. All chemical constituents
were tested for acetylcholinesterase inhibitory activity using the Ellman colorinetric method. The results indicated that
compound 11 exhibited inhibitory activity against acetylcholinesterase.
Key words:Amoora tetrapetala;chemical constituents;acetylcholinesterase inhibitory activity
四瓣崖摩( Amoora tetrapetala Pellegr)是楝科
(Meliaceae)崖摩属(Amoora)植物。崖摩属植物全
世界约 25 ~ 30 种,主要分布于印度、马来半岛一带,
自印度至伊里安岛。我国产 6 ~ 7 种,见于西南和华
南地区[1]。四瓣崖摩为越南及我国滇、黔、桂、粤广
布外,其余多为区域特有或狭域特有,多分布于季雨
林或雨林中[2]。文献报道同属植物铁椤 ( Amoora
tsangii Merr. )的枝和干在民间常用来杀虫[3],其树
皮煎汁可驱虱[4];四瓣崖摩乙醇提取物对小菜蛾成
虫的产卵忌避作用明显[5],但对其化学成分的研究
报道较少。为进一步研究和利用崖摩属植物,本课
题组对四瓣崖摩树枝进行化学成分研究,从中分离
鉴定出 14 个化合物,运用波谱学方法分别鉴定为:
22ξ-hydroxytirucalla-7,24-dien-3,23-dione(1)、dyma-
crin D(2)、泽屋萜(3)、α-菠菜甾醇(4)、8-hydroxy-
6-methoxy-3-pentylisocoumarin ( 5 )、 lichenxanthone
(6)、10-oxo-isodauc-3-en-15-al(7)、( + )-ent-ficusol
(8)、3-甲氧基-4-羟基苯甲酸甲酯(9)、香草醛(10)、
2,4-二羟基-3,6-二甲基苯甲酸甲酯(11)、2-羟基-4-
甲氧基-6-丙基苯甲酸甲酯(12)、2,4-二羟基-6-甲基
苯甲酸甲酯(13)和琥珀酸甲酯(14)。以上化合物
均为首次从崖摩属植物中分离得到。
1 仪器与材料
薄层层析硅胶和柱色谱硅胶(200 ~ 300 目)和
硅胶 H为青岛海洋化工厂产品;Sephadex LH-20 为
Merck公司产品;ODS(20 ~ 45 μm)为 Fuji 公司产
品;MS 谱在 Autospec-3000 质谱仪上测定;NMR 用
Bruker AV-500 型超导核磁仪测定,以 TMS 为内标;
旋光仪为 Rudolph Research Analytical 生产的 Ru-
dolph Autopol Ⅲ polarimeter 型旋光仪;乙酰胆碱酯
酶、碘化硫代乙酰胆碱、二硫代二硝基苯甲酸
(DNTB)、他克林均购自 Sigma 公司;ELX-800 酶标
仪购自美国宝特公司;超净工作台为上海博讯实业
有限公司医疗设备厂。
四瓣崖摩树枝于 2013 年 9 月采集于海南省白
沙县,经中国热带农业科学院热带生物技术研究所
刘寿柏博士鉴定为四瓣崖摩 Amoora tetrapetala Pel-
legr,凭证标本(SBYM201309)存放于中国热带农业
科学院热带生物技术研究所。
2 提取与分离
四瓣崖摩树枝(7. 5 kg)用 95%乙醇室温浸取
三次,每次 7 d,减压浓缩,得乙醇提取物 315. 8 g,将
提取物分散于水中成悬浊液,用石油醚、乙酸乙酯、
正丁醇萃取,得石油醚萃取物 35. 0 g,乙酸乙酯萃取
物 75. 8 g,正丁醇萃取物 31. 8 g。乙酸乙酯部分采
用硅胶(硅胶 H)减压柱,以石油醚-乙酸乙酯(0 ∶ 1
~ 1∶ 0)梯度洗脱,分段收集得到 8 个流分(Fr. 1 ~
8)。Fr. 2(1. 2 g)经 ODS(甲醇-水 3∶ 1 ~ 1∶ 0)梯度
洗脱,得 8 个流分(Fr. 2-1 ~ Fr. 2-8)。Fr. 2-2(69. 0
mg)经反复硅胶柱(200 ~ 300 目)色谱,得化合物 12
(23. 4 mg)、6(1. 8 mg);Fr. 2-3(56. 2 mg)经 Sepha-
dex LH-20 (氯仿-甲醇 1 ∶ 1),得到化合物 5 (1. 2
mg)。Fr. 3(5. 2 g)经 ODS(甲醇-水 1∶ 1 ~ 1∶ 0)梯度
洗脱,得到 20 个流分(Fr. 3-1 ~ Fr. 3-20)。再经过
反复 Sephadex LH-20(氯仿-甲醇 1 ∶ 1)色谱以及硅
胶柱色谱得到化合物 10(7. 1 mg)、9(3. 1 mg)、11
(208. 0 mg)、7(5. 1mg)、4(6. 3 mg)、2(56. 6 mg)、1
(4. 2 mg)、13(1. 4 mg)。Fr. 4(13. 6 g)经 ODS(甲
醇-水 1∶ 1 ~ 1∶ 0)梯度洗脱,得到 10 个流分(Fr. 4-1
~ Fr. 4-10)。Fr. 4-5 (62. 8 mg)经 Sephadex LH-20
(氯仿-甲醇 1 ∶ 1)色谱,结晶得到化合物 3 (14. 7
mg)。Fr. 5(19. 8 g)经 ODS(甲醇-水 3∶ 7 ~ 1∶ 0)梯
度洗脱,得到 18 个流分(Fr. 5-1 ~ Fr. 5-18)。再经
过反复的 Sephadex LH-20(氯仿-甲醇 1 ∶ 1)色谱以
及硅胶柱色谱得到化合物 14 (9. 9 mg)、8 (12. 5
mg)。
3 结构鉴定
化合物 1 白色针晶;[α]25D + 37 ( c 0. 3,
CHCl3);ESI-MS m/z 477[M + Na]
+,453 [M-H]-;
结合 NMR谱和 ESI-MS 数据推断分子式为 C30 H46
O3;
1H NMR (500 MHz,CD3COCD3 ) δ:6. 35 (1H,
m,H-24),5. 36 (1H,m,H-7),4. 16 (1H,m,H-22),
2. 74 (2H,dd,J = 5. 5,14. 6 Hz,H-2),2. 18 (3H,
d,J = 0. 8 Hz,H-27),1. 96 (1H,m,H-1a),1. 41
(1H,m,H-1b),1. 95 (3H,d,J = 1. 0 Hz,H-26),
1. 82 ( 1H,m,H-20 ),1. 10 ( 3H,s,H-30 ),1. 08
(3H,s,H-29),1. 05 (3H,s,H-28),1. 00 (3H,s,H-
19),0. 89 (3H,s,H-18),0. 64 (3H,d,J = 6. 4 Hz,
H-21);13C NMR (125 MHz,CD3COCD3) δ:39. 0 (C-
1),35. 3 ( C-2 ),215. 3 ( C-3 ),48. 2 ( C-4 ),53. 0
(C-5),25. 0 ( C-6),118. 8 ( C-7),146. 7 ( C-8 ),
49. 8 (C-9),35. 7 (C-10),18. 9 (C-11),34. 4 (C-
12),44. 0 (C-13),52. 2 (C-14),34. 7 (C-15),28. 0
(C-16),50. 0 (C-17),13. 0 (C-18),21. 8 (C-19),
40. 0 (C-20),12. 6 (C-21),79. 5 (C-22),201. 2 (C-
23),120. 4 (C-24),158. 9 (C-25),25. 1 (C-26),
21. 1 (C-27),28. 3 (C-28),22. 3 (C-29),27. 8 (C-
30)。以上波谱数据与文献[6]对照基本一致,故鉴
定为 22ξ-hydroxytirucalla-7,24-dien-3,23-dione。
化合物 2 白色针状结晶;[α]25D -36 ( c 1,
CHCl3);ESI-MS m/z 461[M + Na]
+,437 [M-H]-;
结合 NMR谱和 ESI-MS 数据推断分子式为 C30 H46
O2;
1H MNR (500 MHz,CDCl3 ) δ:6. 06 (1H,s,H-
24),5. 31 (1H,br s,H-7),2. 76 (1H,m,H-2a),
2. 27 (1H,m,H-2b),2. 47 (1H,m,H-22a),2. 10
(1H,m,H-22b),2. 29 (1H,m,H-9),2. 14 (3H,s,
H-27),1. 89 (3H,s,H-26),1. 12 (3H,s,H-29),
1. 05 (3H,s,H-28),1. 01 (3H,s,H-19),1. 01 (3H,
s,H-30),0. 89 (3H,d,J = 6. 3 Hz,H-21),0. 86
(3H,s,H-18);13 C MNR (125 MHz,CDCl3) δ:38. 6
(C-1),35. 0 (C-2),217. 0 (C-3),48. 0 (C-4),53. 3
(C-5),24. 5 ( C-6),118. 0 ( C-7),145. 9 ( C-8 ),
48. 5 (C-9),35. 1 (C-10),18. 3 (C-11),33. 6 (C-
365Vol. 27 米承能等:四瓣崖摩化学成分研究
12),43. 7 (C-13),51. 4 (C-14),34. 1 (C-15),28. 5
(C-16),52. 4 (C-17),22. 1 (C-18),12. 9 (C-19),
33. 6 (C-20),19. 5 (C-21),51. 7 (C-22),201. 6 (C-
23),124. 4 (C-24),154. 9 (C-25),27. 8 (C-26),
20. 8 (C-27),24. 7 (C-28),21. 7 (C-29),27. 5 (C-
30)。以上波谱数据与文献[7]对照基本一致,故鉴
定为 dymacrin D。
化合物 3 白色粉末;ESI-MS m/z 467 [M +
Na]+,443[M-H]-;结合 NMR 谱和 ESI-MS 数据推
断分子式为 C30 H52 O2;
1H NMR (500 MHz,DMSO-
d6) δ:3. 96 (1H,d,J = 6. 5 Hz,6-OH),3. 90 (1H,s,
22-OH),3. 73 (2H,m,H-6),2. 08 (1H,m,H-21),
1. 92 (1H,s,H-16a),1. 55 (1H,s,H-16b),1. 61
(1H,m,H-20a),1. 43 (1H,m,H-20b),1. 56 (1H,
m,H-1a),0. 77 (1H,m,H-1b),1. 49 (1H,m,H-
2a),1. 31 (1H,m,H-2b),1. 46 (1H,s,H-11a),
1. 22 (1H,m,H-11b),1. 42 (1H,s,H-19a),0. 84
(1H,m,H-19b),1. 41 (1H,s,H-12a),1. 34 (1H,s,
H-12b),1. 40 (2H,m,H-7),1. 37 (1H,s,H-15a),
1. 13 ( 1H,s,H-15b),1. 36 ( 1H,m,H-3a ),1. 17
(1H,m,H-3b),1. 35 (1H,s,H-13),1. 34 (1H,s,H-
17),1. 24 (1H,m,H-9),1. 10 (3H,s,H-23),1. 06
(3H,s,H-30),1. 02 (3H,s,H-29),0. 96 (3H,s,H-
26),0. 93 (3H,s,H-24),0. 90 (3H,s,H-27),0. 80
(3H,s,H-25),0. 71 (1H,d,J = 10. 8 Hz,H-5),
0. 69 (3H,s,H-28);13C NMR (125 MHz,DMSO-d6)
δ:38. 9 (C-1),18. 2 (C-2),44. 8 (C-3),33. 5 (C-
4),60. 2 (C-5),66. 8 (C-6),44. 8 (C-7),42. 3 (C-
8),49. 5 (C-9),38. 8 (C-10),20. 8 (C-11),23. 8
(C-12),49. 1 (C-13),41. 6 (C-14),34. 1 (C-15),
21. 4 (C-16),54. 0 (C-17),43. 8 (C-18),41. 1 (C-
19),26. 3 (C-20),50. 5 (C-21),71. 9 (C-22),36. 8
(C-23),22. 2 (C-24),17. 1 (C-25),18. 2 (C-26),
17. 0 (C-27),16. 1 (C-28),29. 0 (C-29),31. 0 (C-
30)。以上波谱数据与文献[8]对照基本一致,故鉴
定为泽屋萜(zeorin)。
化合物 4 白色针晶;ESI-MS m/z 435 [M +
Na]+,411[M-H]-;结合 NMR 谱和 ESI-MS 数据推
断分子式为 C29H48O;
1H NMR (500 MHz,CDCl3) δ:
5. 16 (1H,s,H-7),5. 14 (1H,m,H-22),5. 02 (1H,
m,H-23),3. 60 (1H,m,H-3),1. 53 (1H,s,OH),
1. 02 (3H,m),0. 85 (3H,m),0. 81 (6H,d,J = 7. 3
Hz),0. 80 (3H,s),0. 55 (3H,s,H-18);13 C NMR
(125 MHz,CDCl3) δ:37. 3 (C-1),31. 6 (C-2),71. 2
(C-3),38. 1 (C-4),40. 4 (C-5),29. 8 (C-6),117. 6
(C-7),139. 7 ( C-8),49. 6 ( C-9),34. 4 ( C-10 ),
21. 7 (C-11),39. 6 (C-12),43. 4 (C-13),55. 3 (C-
14),23. 2 (C-15),28. 7 (C-16),56. 0 (C-17),12. 2
(C-18),13. 2 (C-19),41. 0 (C-20),21. 3 (C-21),
138. 3 (C-22),129. 6 (C-23),51. 4 ( C-24),32. 0
(C-25),19. 1 (C-26),21. 5 (C-27),25. 6 (C-28),
12. 4 (C-29)。以上波谱数据与文献[9]对照基本一
致,故鉴定为 α-菠菜甾醇(α-spinasterol)。
化合物 5 无色针晶;ESI-MS m/z 285 [M +
Na]+,261[M-H]-;结合 NMR 谱和 ESI-MS 数据推
断分子式为 C15 H18 O4;
1H NMR (500 MHz,CDCl3 )
δ:11. 15 (1H,s,8-OH),6. 47 (1H,s,H-4),6. 54
(1H,d,J = 2. 3 Hz,H-5),6. 46 (1H,d,J = 2. 3
Hz,H-7),3. 90 (3H,s,6-OCH3 ),2. 53 (2H,t,J =
7. 5 Hz,H-1),1. 69 (2H,m,H-2),1. 38 (2H,m,H-
3),1. 36 (2H,m,H-4),0. 90 (3H,m,5-CH3);
13 C
NMR (125 MHz,CDCl3) δ:167. 9 (C-1),159. 0 (C-
3),104. 7 ( C-4 ),140. 8 ( C-4a),101. 8 ( C-5 ),
167. 1 (C-6),100. 9 (C-7),164. 4 (C-8),100. 5 (C-
8a),56. 3 (OCH3),33. 6 (C-1),27. 2 (C-2),31. 8
(C-3),23. 0 (C-4),14. 2 (C-5)。以上波谱数据
与文献[10]对照基本一致,故鉴定为 8-hydroxy-6-me-
thoxy-3-pentylisocoumarin。
化合物 6 浅黄色结晶;ESI-MS m/z 309[M +
Na]+,285[M-H]-;结合 NMR 谱和 ESI-MS 数据推
断分子式为 C16 H14 O5;
1H NMR (500 MHz,CDCl3 )
δ:13. 39 (1H,s,1-OH),6. 69 (1H,d,J = 2. 4 Hz,
H-5),6. 67 (1H,d,J = 2. 4 Hz,H-7),6. 33 (1H,d,
J = 2. 3 Hz,H-4),6. 30 (1H,d,J = 2. 3 Hz,H-2),
3. 89 (3H,s,6-OCH3),3. 87 (3H,s,3-OCH3),2. 85
(3H,s,8-CH3 );
13 C NMR ( 125 MHz,CDCl3 ) δ:
163. 9 (C-1),96. 9 (C-2),166. 0 (C-3),92. 2 (C-
4),98. 6 (C-5),163. 9 (C-6),115. 6 (C-7),143. 6
(C-8),182. 6 (C-9),157. 1 (C-4a),159. 6 (C-4b),
113. 1 (C-8a),104. 3 (C-8b),55. 9 (3-OCH3),55. 8
(6-OCH3),23. 6 (CH3 )。以上波谱数据与文献
11]
对照基本一致,故鉴定为 lichenxanthone。
化合物 7 无色油状物;ESI-MS m/z 257[M +
Na]+,233[M-H]-;结合 NMR 谱和 ESI-MS 数据推
断分 子 式 为 C15 H22 O2;
1H NMR ( 500 MHz,
CD3COCD3) δ:9. 39 (1H,s,H-15),6. 80 (1H,d,J
465 天然产物研究与开发 Vol. 27
= 5. 5 Hz,H-4),1. 69 (1H,m,H-11),1. 34 (3H,s,
H-14),0. 95 (3H,s,H-12),0. 94 (3H,s,H-13);13 C
NMR (125 MHz,CD3COCD3 ) δ:27. 6 (C-1),20. 0
(C-2),144. 9 ( C-3),159. 6 ( C-4),53. 6 ( C-5 ),
56. 1 (C-6),35. 8 (C-7),39. 6 (C-8),60. 3 (C-9),
211. 6 (C-10),33. 3 (C-11),19. 9 (C-12),22. 2 (C-
13),25. 1 (C-14),193. 3 (C-15)。以上波谱数据与
文献[12]对照基本一致,故鉴定为 10-oxo-isodauc-3-
en-15-al。
化合物 8 白色针状结晶;[α]25D + 129 ( c
0. 02,CHCl3);ESI-MS m/z 249[M + Na]
+,225[M-
H]-;结合 NMR谱和 ESI-MS数据推断分子式为 C11
H14 O5;
1H MNR ( 500 MHz,CD3COCD3 ) δ: 6. 92
(1H,m,H-2),6. 75 (1H,m,H-6),6. 77 (1H,m,
H-5),4. 04 (1H,m,H-3b),3. 82 (3H,s,3-OCH3),
3. 70 (1H,dd,J = 3. 4,8. 5 Hz,H-2),3. 67 (1H,m,
H-3a),3. 63 (3H,s,1-OCH3);
13C NMR (125 MHz,
CD3COCD3) δ:174. 0 (C-1),55. 0 (C-2),65. 3 (C-
3),128. 6 ( C-1),112. 5 ( C-2),148. 3 ( C-3),
146. 9 ( C-4),115. 8 ( C-5),121. 5 ( C-6),51. 9
(1-OCH3),56. 2 (3-OCH3 )。以上波谱数据与文
献[13]对照基本一致,故鉴定为( + )-ent-ficusol。
化合物 9 无色油状物;ESI-MS m/z 205[M +
Na]+,181[M-H]-;结合 NMR 谱和 ESI-MS 数据推
断分子式为 C9H10O4;
1H NMR (500 MHz,CDCl3) δ:
7. 63 (1H,dd,J = 1. 7,8. 3 Hz,H-6),7. 55 (1H,d,
J = 1. 6 Hz,H-2),6. 94 (1H,d,J = 8. 3 Hz,H-5),
3. 96 (3H,s,COOCH3),3. 89 (3H,s,3-OCH3 );
13 C
NMR (125 MHz,CDCl3) δ:122. 4 (C-1),111. 8 (C-
2),150. 1 (C-3),146. 3 (C-4),114. 2 (C-5),124. 3
(C-6),167. 0 ( C-7),52. 1 ( COOCH3 ),56. 2 (3-
OCH3)。以上波谱数据与文献
[14]对照基本一致,故
鉴定为 3-甲氧基-4-羟基苯甲酸甲酯(3-methoxy-4-
hydroxybenzoate)。
化合物 10 棕黄色油状物;ESI-MS m/z 175[M
+ Na]+,151[M-H]-;结合 NMR 谱和 ESI-MS 数据
推断分子式为 C8H8O3;
1H NMR (500 Hz,CDCl3) δ:
9. 82 (1H,s,CHO),7. 43 (1H,m,H-6),7. 42 (1H,
s,H-2),7. 04 (1H,d,J = 8. 5 Hz,H-5),6. 22 (1H,
s,OH),3. 96 (3H,s,OCH3);
13 C NMR (125 MHz,
CDCl3) δ:130. 0 (C-1),108. 9 (C-2),147. 3 (C-3),
151. 8 ( C-4 ),114. 5 ( C-5 ),127. 7 ( C-6 ),191. 1
(CHO),56. 3 (OCH3)。以上波谱数据与文献
[15]对
照基本一致,故鉴定为香草醛(vanillin)。
化合物 11 无色针晶;ESI-MS m/z 219 [M +
Na]+,195[M-H]-;结合 NMR 谱和 ESI-MS 数据推
断分 子 式 为 C10 H12 O4;
1H NMR ( 500 MHz,
CD3COCD3) δ:12. 01 (1H,s,2-OH),6. 32 (1H,s,
H-5),3. 90 (3H,s,OCH3 ),2. 41 (3H,s,6-CH3 ),
2. 03 ( 3H,s,3-CH3 );
13 C NMR ( 125 MHz,
CD3COCD3) δ:104. 9 (C-1),160. 9 ( C-2),109. 5
(C-3),164. 1 (C-4),111. 4 (C-5),140. 6 (C-6),
173. 5 (C = O),52. 1 (OCH3 ),8. 1 (3-CH3 ),24. 2
(6-CH3)。以上波谱数据与文献
[16]对照基本一致,
故鉴定为 2,4-二羟基-3,6-二甲基苯甲酸甲酯
(methyl 2,4-hydroxy-3,6-dimethylbenzoate)。
化合物 12 黄色油状物;ESI-MS m/z 247[M +
Na]+,223[M-H]-;结合 NMR 谱和 ESI-MS 数据推
断分子式为 C12 H16 O4;
1H NMR (500 MHz,CDCl3 )
δ:11. 74 (1H,s,OH),6. 33 (1H,d,J = 2. 6 Hz,H-
3),6. 28 (1H,d,J = 2. 6 Hz,H-5),3. 92 (3H,s,
COOCH3),3. 79 (3H,s,4-OCH3),2. 82 (2H,t,J =
7. 7 Hz,H-1),1. 55 (2H,m,H-2),0. 95 (3H,t,J =
7. 4 Hz,H-3);13C NMR (125 MHz,CDCl3) δ:104. 7
(C-1),165. 7 ( C-2),98. 9 ( C-3),164. 0 ( C-4 ),
110. 8 (C-5),147. 8 (C-6),39. 0 (C-1),25. 0 (C-
2),14. 4 (C-3),172. 1 (C = O),52. 0 (COOCH3),
55. 4 (4-OCH3)。以上波谱数据与文献
[17]对照基本
一致,故鉴定为 2-羟基-4-甲氧基-6-丙基苯甲酸甲酯
(methyl 2-hydroxy-4-methoxy-6-propylbenzoate)。
化合物 13 棕黄色无定形固体;ESI-MS m/z
205[M + Na]+,181 [M-H]-;结合 NMR 谱和 ESI-
MS 数据推断分子式为 C9H10 O4;
1H NMR ( 500
MHz,CDCl3) δ:6. 30 (1H,d,J = 2. 5 Hz,H-3),
6. 25 (1H,d,J = 2. 0 Hz,H-5),3. 95 (3H,s,OCH3),
2. 51 (3H,s,CH3);
13 C NMR (125 MHz,CDCl3) δ:
144. 1 (C-1),165. 5 (C-2),101. 4 (C-3),160. 5 (C-
4),111. 5 ( C-5),105. 8 ( C-6),172. 3 ( C = O),
52. 0 (OCH3 ),24. 5 (CH3 )。以上波谱数据与文
献[18]对照基本一致,故鉴定为 2,4-二羟基-6-甲基
苯甲酸甲酯 ( methyl 2,4-dihydroxy-6-methylbenzo-
ate)。
化合物 14 无色油状物;ESI-MS m/z 155[M +
Na]+,131[M-H]-;结合 NMR 谱和 ESI-MS 数据推
565Vol. 27 米承能等:四瓣崖摩化学成分研究
断分子式为 C5H8O4;
1H NMR (500 MHz,CDCl3) δ:
3. 70 (3H,s,OCH3),2. 67 (2H,m,H-2),2. 63 (2H,
m,H-3);13 C NMR (125 MHz,CDCl3 ) δ:29. 0 (C-
2),28. 8 ( C-3),177. 6 ( COOH),172. 8 ( COO-),
52. 1 (OCH3);以上波谱数据与文献
[19]对照基本一
致,故鉴定为琥珀酸甲酯(methyl hydrogen succi-
nate)。
4 活性检测
测定化合物抗乙酰胆碱酯酶活性方法参照 El-
man[20]。待测样品均用 DMSO 进行溶解。取 110
μL磷酸缓冲液(pH 8. 0),10 μL待测样品(100 μg /
mL)和 40 μL乙酰胆碱酯酶(0. 02 μg /mL)于 96 孔
板中,温育 20 min(30 ℃),之后加入 DTNB(2. 48
mg /mL)和碘化硫代乙酰胆碱(1. 81 mg /mL)等体积
混合液 40 μL,反应体系总共 200 μL,30 min后,405
nm处酶标仪进行检测。阳性对照为他克林,反应终
浓度为 0. 08 μg /mL,阴性对照为 DMSO,反应终浓
度为 0. 1%,实验 3 次重复。按照(E-S) /E × 100%
来计算化合物对乙酰胆碱酯酶的抑制率(E 为阴性
对照平均吸光值,S为待测样品的平均吸光值)。最
终测得阴性对照的平均吸光值为 0. 9790,阳性对照
的平均吸光值为 0. 5618,化合物 11 的平均吸光值
为 0. 7797,可以得到化合物 11 对乙酰胆碱酯酶的
抑制率为 20. 36%,阳性对照(他克林)的抑制率为
42. 61%,说明化合物 11 对乙酰胆碱酯酶有一定的
抑制作用。
5 结果与讨论
本文采取多种色谱技术,从四瓣崖摩中分离得
到了 14 个化合物,均为首次从崖摩属植物中分离得
到。在本次研究中我们发现化合物 11 对乙酰胆碱
酯酶具有抑制活性,推断该化合物可能具有神经保
护的作用。同时,文献报道化合物 3 具有抗结核活
性和抗 HIV 活性[8],化合物 6 对淋巴白血病 P388、
胰腺癌细胞 BXPC-3、乳腺癌细胞 MCF-7 等具有良
好的抗癌、抗肿瘤活性[21],化合物 8 具有抗结核活
性[19]。研究结果丰富了崖摩属植物化学成分和生
物活性的多样性,为研究和利用该属植物提供了科
学依据。
参考文献
1 Yunnan Institute of Botany (云南植物研究所) . Flora Yun-
nanica (云南植物志) . Beijing: Science and Technology
Press (北京科学技术出版社),1977. 231.
2 Wu ZY (吴征镒),Lu AM (路安民),Tang YC (汤彦承),
et al. Review of the genus China angiosperm families (中国
被子植物科属综论) . Beijing:Science Press (北京科学出
版社),2003. 746.
3 Hou KZ (候宽昭),Chen DZ (陈德昭) . Flora of China (中
国植物志) . Beijing:Science Press (北京科学出版社),
1997.
4 South China Institute of Botany (中国科学院华南植物研究
所) . Flora Hainanica (海南植物志) . Beijing:Science Press
(北京科学出版社),1974. 3,67.
5 Li LF (黎柳锋),Zeng T (曾涛),Wei DW (韦德卫),et
al. Effect of different plant extracts on prevention of oviposi-
tion in Plutella xylostella (L. ) adults. J Southern Agric (南
方农业学报),2011,42:155-157.
6 Liang GY,Gray AI,Waterman PC. Tirucallane and oleanane
triterpenes from the resin of Aucoumea klaineana. Phytochem-
istry,1988,27:2283-2286.
7 Mohamad K,Martin MT,Litaudon M,et al. Tirucallane triter-
penes from Dysoxylum macranthum. Phytochemistry,1999,
52:1461-1468.
8 Knig GM,Wright AD. 1H and 13C NMR and biological ac-
tivity investigations of four lichen-derived compounds. Phyto-
chem Anal,1999,10:279-284.
9 Zhao XY (赵晓亚),Sun HD (孙汉董),Wu JZ (吴继洲) .
Studies on chemical constituents from rhizome of Impatien
pritzellii var. hupehensis. China J Chin Mater Med (中国中
药杂志),2005,30:584-586.
10 Kihampa C,Nkunya MHH,Joseph CC,et al. Anti-mosquito
and antimicrobial nor-halimanoids,isocoumarins and an ani-
linoid from Tessmannia densiflora. Phytochemistry,2009,70:
1233-1238.
11 Yang JX (杨建香),Qiu SX (邱声祥),She ZG (佘志刚),
et al. Metabolites of mangrove endophytic fungus SK7RN3G1
from south China sea. Chin J Exp Tradit Med Form (中国实
验方剂学杂志),2012,18(21):95-98.
12 Hou L (侯丽),Tang GH (唐贵华),Zhang Y (张于),et
al. A new carotane sesquiterpene from Walsura robusta. Chin
J Nat Med (中国天然药物),2013,11:84-86.
13 Kuo HT,Peng CF,Huang HY,et al. Chemical constituents
and antitubercular activity of formosan Pisonia umbellifera.
Planta Med,2011,77:736-741.
14 Yoshioka T,Inokuchi T,Fujioka S,et al. Phenolic compounds
and flavonoids as plant growth regulators from fruit and leaf
of Vitex rotundifolia. Z Naturforsch,2004,59:509-514.
(下转第 584 页)
665 天然产物研究与开发 Vol. 27
3:458-463.
5 Jian Z (坚哲) . The role of Nrf2-ARE pathway in the oxida-
tive stress pathogenesis of vitiligo and its mechanism. Xian:
The Fourth Military Medical University(第四军医大学),
PhD. 2013.
6 Yu R,Lei W,Mandlekar S,et al. Role of a mitogen-activated
protein kinase pathway in the induction of phase II detoxif-
ying enzymes by chemicals. J Biol Chem,1999,274:27545-
27552.
7 Zhang RC (张瑞晨),Liu B (刘斌),Sun ZX (孙震晓),et
al. Effects of extract of Polygonum multiflorum on cell cycle
arrest and apoptosis of human liver cell line L02. J Chin Inte-
gr Med(中西医结合学报),2010,8:554-561.
8 Zheng BS (郑必胜),Tang FY (唐芳勇) . Progress in re-
search on polysaccharides from Polygonum Mulitiflorum
Thunb. . Acade Periodic Farm Prod Proc(农产品加工·学
刊),2008,2(127):30-32.
9 Luo ZX (罗增香),Xiang LH (项蕾红),Li J (李剑),et
al. Effects of two Chinese medicine monomers on the tyrosi-
nase activity and melanin synthesis in both pure human cul-
tured melanocytes and co-culture system of melanocytes and
keratinocytes in vitro. China J Leprosy Skin Dis(中国麻风皮
肤病杂志),2008,8:583-585.
10 Liu R (刘荣),Sun JN (孙建宁),Guo YJ (郭亚健) . As-
sessment of whitening efficacy of cosmetical materials in B16
melanoma cells. Chin J Aesthetic Med (中国美容医学),
2011,7:1114-1117.
11 Kim J,Kim JH,Jeung ES. Inhibitory effects of stilbene gluco-
side isolated from the root of Polygonum multiflorum on ty-
rosinase activity and melanin biosynthesis. J Korean Soc Appl
Biol Chem,2009,52:342-345.
12 Jiang ZQ (姜泽群),Wu Q (吴琼),Xu JM (徐继敏),et
al. Study on the mechanism of Polygonum promote the forma-
tion of melanin. J Nanjing Univ Tradit Chinese Med(南京中
医药大学学报),2010,03:190-192.
13 Jiang ZQ,Xu JM,Long MH,et al. 2,3,5,4-tetrahydroxystil-
bene-2-O-β-D-glucoside (THSG) induces melanogenesis in
B16 cells by MAP kinase activation and tyrosinase upregula-
tion. Life Sci,2009,85:345-350.
14 Emerit I,Filipe P,Freitas J,et al. Protective effect of super-
oxide dismutase against hair graying in a mouse model. Pho-
tochem Photobiol,2004,80:579-582.
15 Zhang MS (张绵松),Liu X (刘新),Meng XM (孟秀梅),
et al. Antioxidative and antimicrobial activities of Polygonum
multiflorum Thunb. in vitro. Food Sci Technol(食品科技),
2012,37:228-231.
16 Lv LS (吕丽爽),Tang J (汤坚),He QT (何淇傥) . Prop-
erty and activity of DPPH radical scavenging of stilbene glu-
cosides. J Harbin Instit Technol(哈尔滨工业大学学报),
2009,41:
櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵櫵
252-255.
(上接第 566 页)
15 Song ZZ (宋治中),Jia ZJ (贾忠建),Zhu QX (朱启秀) .
Studies on the chemical components of Bupleurum sibiricum
Vest (Ⅱ) . Chem J Chinese Uinv (高等学校化学学报),
1991,12:1469-1472.
16 Zheng WP (郑卫平),Tang YP (唐于平),Lou FC (楼凤
昌),et al. Studies on the constituents of Dendrobium chryse-
um Rolfe. J Chin Pharm Univ (中国药科大学学报),2000,
31:5-7.
17 Lubbe M,Gütlein JP,Reinke H,et al. Regioselective synthe-
sis of polyketide-type phenols by formal [3 + 3]-cyclocon-
densations of 1,3-bis ( silyloxy )-1,3-butadienes with 3-
oxoorthoesters. Synlett,2008,17:2671-2673.
18 Guo HL,Lei L,Meng D,et al. The chemical constituents of
the fungus Stereum sp. . Chem Biodivers,2006,3:210-216.
19 Xiong Y (熊英),Deng KZ (邓可众),Gao WY (高文远),
et al. Studies on chemical constituents of Ranunculus terna-
tus. China J Chin Mater Med (中国中药杂志),2008,33:
909-911.
20 Ellman GL,Courtney KD,Andres VJ,et al. A new and rapid
colorimetric determination of acetylcholinesterase activity.
Biochem Pharmacol,1961,7 (2):88-95.
21 Wansi JW,Wandji J,Waffo AFK,et al. Alkaloids from Orici-
opsis glaberrima Engl. ( Rutaceae) . Phytochemistry,2006,
67:475-480.
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