药学学报

No.	1
1		127.6		130.1
2	7.07 (1H, d, J = 2.2 Hz)	110.0	6.76 (1H, s)	105.8
3		149.4		153.6
3-OMe	3.91 (3H, s)	56.1	3.88 (3H, s)	56.4
4		151.6		140.6
4-OMe	3.91 (3H, s)	56.1	3.88 (3H, s)	61.2
5	6.85 (1H, d, J = 8.2 Hz)	111.1		153.6
5-OMe			3.88 (3H, s)	56.4
6	7.10 (1H, dd, J = 8.2, 2.2 Hz)	123.6	6.76 (1H, s)	105.8
1′		124.3		124.2
2′		146.2		146.3
2′-OMe	3.80 (3H, s)	62.2	3.81 (3H, s)	62.2
3′		143.2		143.2
3′-OMe	3.91 (3H, s)	61.4	3.91 (3H, s)	61.5
4′		148.8		148.9
4′-OMe	3.99 (3H, s)	61.6	3.99 (3H, s)	61.6
5′		143.2		143.2
5′-OMe	3.91 (3H, s)	61.4	3.91 (3H, s)	61.5
6′		146.2		146.3
6′-OMe	3.80 (3H, s)	62.2	3.81 (3H, s)	62.2
α	6.88 (1H, d, J = 16.0 Hz)	126.8	6.90 (1H, d, J = 15.9 Hz)	128.1
β	7.27 (1H, d, J = 16.0 Hz)	146.3	7.25 (1H, d, J = 15.9 Hz)	146.0
C=O		193.7		193.5

No.	1
1		127.6		130.1
2	7.07 (1H, d, J = 2.2 Hz)	110.0	6.76 (1H, s)	105.8
3		149.4		153.6
3-OMe	3.91 (3H, s)	56.1	3.88 (3H, s)	56.4
4		151.6		140.6
4-OMe	3.91 (3H, s)	56.1	3.88 (3H, s)	61.2
5	6.85 (1H, d, J = 8.2 Hz)	111.1		153.6
5-OMe			3.88 (3H, s)	56.4
6	7.10 (1H, dd, J = 8.2, 2.2 Hz)	123.6	6.76 (1H, s)	105.8
1′		124.3		124.2
2′		146.2		146.3
2′-OMe	3.80 (3H, s)	62.2	3.81 (3H, s)	62.2
3′		143.2		143.2
3′-OMe	3.91 (3H, s)	61.4	3.91 (3H, s)	61.5
4′		148.8		148.9
4′-OMe	3.99 (3H, s)	61.6	3.99 (3H, s)	61.6
5′		143.2		143.2
5′-OMe	3.91 (3H, s)	61.4	3.91 (3H, s)	61.5
6′		146.2		146.3
6′-OMe	3.80 (3H, s)	62.2	3.81 (3H, s)	62.2
α	6.88 (1H, d, J = 16.0 Hz)	126.8	6.90 (1H, d, J = 15.9 Hz)	128.1
β	7.27 (1H, d, J = 16.0 Hz)	146.3	7.25 (1H, d, J = 15.9 Hz)	146.0
C=O		193.7		193.5

Compound	IC₅₀/μmol·L^-1
2	> 100.	93.65
5	> 100.	> 100.
6	> 100.	> 100.
7	32.55	34.15
8	> 100.	> 100.
9	> 100.	27.64
10	4.19	4.85
11	1.91	0.98
13	59.73	75.72
14	> 100.	> 100.
15	> 100.	> 100.
16	> 100.	82.12
17	> 100.	> 100.

Compound	IC₅₀/μmol·L^-1
2	> 100.	93.65
5	> 100.	> 100.
6	> 100.	> 100.
7	32.55	34.15
8	> 100.	> 100.
9	> 100.	27.64
10	4.19	4.85
11	1.91	0.98
13	59.73	75.72
14	> 100.	> 100.
15	> 100.	> 100.
16	> 100.	82.12
17	> 100.	> 100.

云南野独活的化学成分研究

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刘志莹 ¹ , 葛格 ¹ , 刘庭亮 ² , 郑文琳 ² , 郭亚萍 ² , 王书云 ¹^,^* , 吴炎 ²^,^*

药学学报 | 专题报道: 以多学科交叉探寻中药现代化发展之路 2025,60(3): 646-654

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药学学报 | 专题报道: 以多学科交叉探寻中药现代化发展之路 2025, 60(3): 646-654

云南野独活的化学成分研究

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刘志莹¹, 葛格¹, 刘庭亮², 郑文琳², 郭亚萍², 王书云¹^,^*, 吴炎²^,^*

作者信息

1.河南大学药学院, 河南开封 475004

2.深圳技术大学药学院, 广东深圳 518118

通讯作者:

^*王书云, E-mail: wangshuyun0426@163.com

吴炎, E-mail: wuyan1219@yeah.net

Chemical constituents from Miliusa tenuistipitata

Zhi-ying LIU¹, Ge GE¹, Ting-liang LIU², Wen-lin ZHENG², Ya-ping GUO², Shu-yun WANG¹^,^*, Yan WU²^,^*

Affiliations

1. School of Pharmacy, Henan University, Kaifeng 475004, China

2. College of Pharmacy, Shenzhen Technology University, Shenzhen 518118, China

出版时间: 2025-03-12 doi: 10.16438/j.0513-4870.2024-0772

文章导航

摘要

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采用硅胶柱色谱、LH-20葡聚糖凝胶柱色谱及半制备液相等色谱分离技术, 从云南野独活(Miliusa tenuistipitata W. T. Wang) 枝叶的95%乙醇提取物中分离得到20个化合物。通过多种现代波谱学方法鉴定化合物结构, 分别为tenuistone L (1)、tenuistone K (2)、樱花素(3)、藿香黄酮醇(4)、雷杜辛黄酮醇(5)、垂叶黄素(6)、蔓荆子黄素(7)、3, 7-二甲氧基-5, 3′, 4′-三羟基黄酮(8)、泽兰黄醇(9)、(+) -miliusol (10)、miliusane XIX (11)、miliusane XVIII (12)、miliusolide (13)、19, 20-dihydromiliusolide (14)、debilisone C (15)、goniothalamusin (16)、2, 10-dihydroxy-3, 9-dimethoxy-8-oxo-protoberberine (17)、consanguine B (18)、6-羟基豆甾-4, 22-二烯-3-酮(19) 和6-羟基豆甾-4-烯-3-酮(20), 其中化合物1和2为新化合物, 化合物6、9、18和20为首次从番荔枝科植物中分离得到, 化合物5、7、17和19为首次从野独活属植物中分离得到。采用MTT法进行抗肿瘤活性测试, 结果发现化合物10和11可以显著抑制MDA-MB-231和BT-549细胞增殖, 其半数抑制浓度(IC₅₀) 为0.98~4.85 μmol·L^-1。

关键词

云南野独活 / 番荔枝科 / 化学成分 / 抗肿瘤活性

Abstract

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Twenty compounds were isolated from the twigs and leaves of the 95% ethanol extract of Miliusa tenuistipitata W. T. Wang by various chromatographic techniques including silica gel column chromatography, sephadex LH-20 and preparative HPLC. Their structures were determined by analysis of spectral data as tenuistone L (1), tenuistone K (2), sakuranetin (3), pachypodol (4), retusin (5), penduletin (6), casticine (7), 3, 7-dimetoxi-5, 3′, 4′-tri-hidroxiflavona (8), eupatin (9), (+)-miliusol (10), miliusane XIX (11), miliusane XVIII (12), miliusolide (13), 19, 20-dihydromiliusolide (14), debilisone C (15), goniothalamusin (16), 2, 10-dihydroxy-3, 9-dimethoxy-8-oxo-protoberberine (17), consanguine B (18), 6β-hydroxystigmasta-4, 22-dien-3-one (19) and 6β-hydroxystigmast-4-en-3-one (20), respectively. Among them, compounds 1 and 2 are new compounds, compounds 6, 9, 18 and 20 are isolated from Annonaceae plants for the first time and compounds 5, 7, 17 and 19 are isolated from the genus Miliusa for the first time. Human triple negative breast cancer cells (MDA-MB-231 and BT-549) were used to evaluate the antitumor activity of these isolates by MTT assay. Compounds 10 and 11 showed significant anti-proliferation activity against the test cells with half inhibition concentration (IC₅₀) of 0.98-4.85 μmol·L^-1.

Key words

Miliusa tenuistipitata / Annonaceae / chemical constituent / antitumor activity

引用本文

刘志莹, 葛格, 刘庭亮, 郑文琳, 郭亚萍, 王书云, 吴炎. 云南野独活的化学成分研究. 药学学报, 2025 , 60 (3) : 646 -654 . DOI: 10.16438/j.0513-4870.2024-0772

Zhi-ying LIU, Ge GE, Ting-liang LIU, Wen-lin ZHENG, Ya-ping GUO, Shu-yun WANG, Yan WU. Chemical constituents from Miliusa tenuistipitata[J]. Acta Pharmaceutica Sinica, 2025 , 60 (3) : 646 -654 . DOI: 10.16438/j.0513-4870.2024-0772

正文

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云南野独活(Miliusa tenuistipitata W. T. Wang) 为番荔枝科(Annonaceae) 野独活属(Miliusa) 植物, 俗称短柄密榴木, 主要分布于贵州晴隆、云南勐海及勐腊、西藏墨脱等地区, 生于海拔1 500 m山地阔叶林中或山谷灌木丛中。野独活属植物民间用于散寒止痛、祛风除湿, 也用于治疗下肢发凉、麻木、疼痛、抽筋等^[1]。该属植物主要含有高龙胆酸衍生物^[2]、黄酮^[3]及生物碱^[4]等化学成分^[5], 具有抗肿瘤^{[6, 7]}、抗炎^{[8, 9]}及抗菌^{[10, 11]}等药理活性。目前对云南野独活的化学成分和药理作用研究较少, 为进一步探索其化学内涵、揭示其活性成分, 本研究对云南野独活枝叶的化学成分进行系统研究, 共从中分离得到20个化合物, 包括2个查尔酮类化合物(1和2), 1个二氢黄酮类化合物(3), 6个黄酮类化合物(4~9), 3个高龙胆酸衍生物(10~12), 4个番荔枝内酯类化合物(13~16), 2个生物碱类化合物(17和18) 和2个甾醇类化合物(19和20), 化合物结构见图 1。其中化合物1和2为新化合物, 化合物6、9、18和20为首次从番荔枝科植物中分离得到, 化合物5、7、17和19为首次从野独活属植物中分离得到。以人三阴性乳腺癌细胞MDA-MB-231和BT-549为供试细胞株, 采用MTT法对部分化合物进行抗肿瘤活性测试, 结果发现, 化合物10和11可以显著抑制供试细胞增殖, 其IC₅₀值为0.98~4.85 μmol·L^-1。

结果与讨论

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1 结构鉴定

化合物1为淡黄色固体, 易溶于甲醇。HR-ESI-MS谱中显示分子离子峰m/z 419.169 6 [M+H]⁺ (计算值为C₂₂H₂₇O₈⁺, 419.170 0), 提示化合物分子式为C₂₂H₂₆O₈, 不饱和度为10。UV光谱(MeOH) 显示在242 (log ε = 2.15) 和340 (log ε = 2.33) nm处有最大吸收, 且带Ⅰ强而带Ⅱ弱, 提示化合物1为查尔酮类化合物。¹H NMR谱中(表 1), δ_H 6.88 (1H, d, J = 16.0 Hz) 和δ_H 7.27 (1H, d, J = 16.0 Hz) 为查尔酮的α和β特征质子信号。此外, ¹H NMR谱中还有: 7个甲氧基质子信号[δ_H 3.99 (3H, s), 3.91 (3H, s), 3.91 (3H, s), 3.91 (3H, s), 3.91 (3H, s), 3.80 (3H, s), 3.80 (3H, s)], 1组ABX耦合系统质子信号[δ_H 7.10 (1H, dd, J = 8.2, 2.2 Hz), 7.07 (1H, d, J = 2.2 Hz), 6.85 (1H, d, J = 8.2 Hz)], 此偶合关系经¹H-¹H COSY谱(图 2) 所证实[H-5 (δ_H 6.85) 与H-6 (δ_H 7.10) 相关]。¹³C NMR谱给出22个碳信号, 包括1个羰基碳信号(δ_C 193.7)、14个sp²杂化碳信号(δ_C 151.6, 149.4, 148.8, 146.3, 146.2, 146.2, 143.2, 143.2, 127.6, 126.8, 124.3, 123.6, 111.1, 110.0) 和7个甲氧基碳信号(δ_C 62.2, 62.2, 61.6, 61.4, 61.4, 56.1, 56.1)。通过与文献^{[12, 13]}数据比对, 进一步确证化合物1为查尔酮类化合物。HMBC谱(图 2) 中, 通过H-2 (δ_H 7.07) 和H-6 (δ_H 7.10) 分别与C-1 (δ_C 127.6)、C-4 (δ_C 151.6)、C-β (δ_C 146.3) 相关, H-5 (δ_H 6.85) 与C-4 (δ_C 151.6) 相关, 以及3-OMe (δ_H 3.91) 与C-3 (δ_C 149.4) 相关和4-OMe (δ_H 3.91) 与C-4 (δ_C 151.6) 相关, 提示ABX耦合系统在B环, 并确定了取代基在B环的位置。HMBC谱中, 通过2′-OMe (δ_H 3.80) 和6′-OMe (δ_H 3.80) 与C-2′ (δ_C 146.2) 和C-6′ (δ_C 146.2) 相关, 3′-OMe (δ_H 3.91) 和5′-OMe (δ_H 3.91) 与C-3′ (δ_C 143.2) 和C-5′ (δ_C 143.2) 相关, 4′-OMe (δ_H 3.99) 与C-4′ (δ_C 148.8) 相关, 可确定A环取代基位置。通过HMBC谱中, H-α (δ_H 6.88) 分别与C-1 (δ_C 127.6)、C-1′ (δ_C 124.3)、羰基碳(δ_C 193.7) 相关, H-β (δ_H 7.27) 分别与C-2 (δ_C 110.0)、C-6 (δ_C 123.6)、羰基碳(δ_C 193.7) 相关, 最终确定化合物的结构。综上所述, 化合物1结构为3, 4, 2′, 3′, 4′, 5′, 6′-heptamethoxychalcone。经文献检索, 化合物1为一未见报道的新化合物, 命名为tenuistone L。

化合物2为淡黄色固体, 易溶于甲醇。HR-ESI-MS谱中显示分子离子峰m/z 449.179 1 [M+H]⁺ (计算值为C₂₃H₂₉O₉⁺, 449.180 6), 提示化合物分子式为C₂₃H₂₈O₉, 不饱和度为10。UV光谱(MeOH) 中显示最大吸收在250 (log ε = 2.74) 和348 (log ε = 3.10) nm处。对比化合物2与化合物1的质谱和核磁数据发现, 化合物2比化合物1多一个甲氧基信号(δ_H 3.88, δ_C 56.4), 且化合物2的H-2 (δ_H 6.76 1H, s) 和H-6 (δ_H 6.76 1H, s) 均为单峰, C-5化学位移向低场移动42.5, 提示该甲氧基位于C-5。此外, HMBC谱(图 2) 中, δ_H 3.88与C-5 (δ_C 153.6) 相关, 进一步证实上述推测。因此, 确定化合物2结构为3, 4, 5, 2′, 3′, 4′, 5′, 6′-octamethoxychalcone。经文献检索, 化合物2为未报道的新化合物, 命名为tenuistone K。

2 抗肿瘤活性研究

采用MTT法, 以人三阴性乳腺癌细胞MDA-MB-231和BT-549为供试细胞株, 对化合物2、5~11、13~17进行抗肿瘤活性测试。结果表明, 化合物10和11可以显著抑制供试细胞增殖, 详细结果如表 2所示。

3 结论

本研究从云南野独活枝叶中分离鉴定到了20个化合物, 其中化合物1和2为新化合物, 化合物6、9、18和20为首次从番荔枝科植物中分离得到, 化合物5、7、17和19为首次从野独活属植物中分离得到。抗肿瘤活性测试发现, 化合物10和11可以显著抑制MDA-MB-231和BT-549细胞增殖, 其IC₅₀值为0.98~4.85 μmol·L^-1。本研究进一步丰富了云南野独活的化学库, 发现了具有显著抗肿瘤细胞增殖的活性化合物, 为该植物的进一步研究奠定了物质基础。

实验部分

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BUCHI Rotavapor R-100旋转蒸发仪(瑞士BUCHI公司); BCE224-1CCN万分之一分析天平[赛多利斯科学仪器(北京) 有限公司]; Bruker AV-400和Bruker AV-600核磁共振波谱仪(德国Bruker公司); ThermoFisher高分辨质谱仪(美国赛默飞世尔科技公司); Agilent 1260和UNIMICRO-EasySep^®-3050型半制备型高效液相色谱仪(美国安捷伦科技公司和上海通微分析技术有限公司); Agilent 1260和ThermoFisher分析型高效液相色谱仪(美国安捷伦科技公司和美国赛默飞世尔科技公司)。GF₂₅₄薄层色谱硅胶板(烟台新诺新材料技术有限公司); 柱层析硅胶(100~200和200~300目, 青岛海洋化工有限公司); Sephadex LH-20柱层析色谱填料(日本Tosoh Bioscience公司); ODS柱层析填料(Spherical C18 Monomeric, 120A, 50 μm, 加拿大SILICYCLE公司); 分析型HPLC色谱柱(Global Chromatography C18, 4.6 mm × 250 mm, 10 μm, 上海通微分析技术有限公司) 和Cosmosil 5C18-MS-Ⅱ (4.6 mm × 250 mm, 5 μm, 日本Nacalai Tesque公司); 半制备HPLC色谱柱(Global Chromatography C18, 10 mm × 250 mm, 10 μm, 上海通微分析技术有限公司) 和Cosmosil 5C18-MS-Ⅱ (10 mm × 250 mm, 5 μm, 日本Nacalai Tesque公司); 所用化学试剂均为色谱纯(安耐吉化学公司) 或分析纯(上海泰坦科技股份有限公司)。

云南野独活植物材料于2022年6月采集于中国云南省, 经深圳技术大学药学院姚大红博士鉴定为番荔枝科野独活属植物云南野独活(Miliusa tenuistipitata W. T. Wang), 植物标本(No. 20220925) 保存于深圳技术大学中药标本室。

1 提取分离

取干燥云南野独活(Miliusa tenuistipitata) 枝叶29 kg, 粉碎后经95%乙醇渗漉提取4次, 合并滤液并减压回收, 得浸膏0.8 kg; 浸膏使用1.5倍硅胶(1.2 kg) 拌样, 依次用石油醚、二氯甲烷和乙酸乙酯各15 L固相萃取, 获得二氯甲烷萃取物150.0 g。二氯甲烷萃取物进一步经硅胶柱色谱分离, 以石油醚-乙酸乙酯为洗脱剂(100∶0~0∶100), 获得7个组分(A1~A7)。

A2 (56.0 g) 经硅胶柱色谱(石油醚-乙酸乙酯, 100∶0~0∶100) 分离, 共获得10个组分A2C1~A2C10。A2C4部分经半制备液相(乙腈-水, 60∶40) 纯化得到化合物11 (t_R = 18 min; 66.9 mg); A2C5 (6.0 g) 部分经ODS (甲醇-水, 60∶40~100∶0) 分离再经半制备液相(乙腈-水, 46∶54) 纯化, 得到化合物5 (t_R = 43 min; 1.0 mg)。A2C7 (20.0 g) 部分经ODS (甲醇-水, 60∶40~100∶0) 分离, 获得12个组分A2C7D1~A2C7D12, A2C7D3经半制备液相(乙腈-水, 45∶55) 纯化得到化合物1 (t_R = 21 min; 8.1 mg) 及3 (t_R = 16 min; 1.0 mg); A2C7D6经半制备液相(乙腈-水, 60∶40) 纯化得到化合物13 (t_R = 58 min; 3.0 mg); A2C7D7经半制备液相(乙腈-水, 65∶35) 纯化得到化合物14 (t_R = 47 min; 5.4 mg); A2C7D8经半制备液相(乙腈-水, 74∶26) 纯化得到化合物15 (t_R = 51 min; 9.0 mg); A2C7D9经半制备液相(乙腈-水, 75∶25) 纯化得到化合物16 (t_R = 42 min; 17.2 mg); A2C7D10经半制备液相(乙腈-水, 85∶15) 纯化得到化合物19 (t_R = 93 min; 4.3 mg) 和20 (t_R = 100 min; 11.5 mg)。A2C8部分经Sephadex LH-20柱(甲醇) 分离后进一步经半制备液相(乙腈-水, 25∶75) 纯化, 得到化合物18 (t_R = 56 min; 2.0 mg)。

A5 (70.0 g) 部分经ODS (甲醇-水, 60∶40~100∶0) 分离, 获得13个组分(A5B1~A5B13)。A5B2 (56.0 g) 再经ODS (甲醇-水, 40∶60~100∶0) 洗脱, 得到8个组分(A5B2C1~A5B2C8), A5B2C6进一步通过硅胶柱色谱(二氯甲烷-甲醇, 100∶1) 纯化得到化合物10 (25.0 g); A5B2C3经Sephadex LH-20柱(甲醇) 分离再经半制备液相(乙腈-水, 28∶72) 纯化, 得到化合物17 (t_R = 18 min; 5.3 mg); A5B2C4经Sephadex LH-20柱(甲醇) 分离后再经半制备液相(甲醇-水, 54∶46) 纯化, 得到化合物9 (t_R = 23 min; 1.0 mg); A5B2C7经Sephadex LH-20柱(甲醇)、半制备液相(甲醇-水, 62∶38) 纯化, 得到化合物12 (t_R = 47 min; 1.9 mg)、7 (t_R = 27 min; 4.0 mg) 和8 (t_R = 33 min; 8.7 mg)。A5B4 (10.6 g) 经ODS (甲醇-水, 40∶60~100∶0) 分离, 得到43个组分(A5B4C1~A5B4C43), A5B4C14经Sephadex LH-20柱(甲醇)、半制备液相(乙腈-水, 50∶50) 纯化, 得到化合物2 (t_R = 21 min; 4.4 mg); A5B4C16经Sephadex LH-20柱(甲醇) 后经半制备液相(乙腈-水, 30∶70) 纯化, 得到化合物6 (t_R = 76 min; 1.3 mg); A5B4C24析出黄色晶体, 经半制备液相(乙腈-水, 50∶50) 纯化, 得到化合物4 (t_R = 27 min; 2.5 mg)。

2 结构鉴定

化合物1 淡黄色固体; HR-ESI-MS m/z 419.169 6 [M+H]⁺ (计算值为C₂₂H₂₇O₈⁺, 419.170 0), 分子式为C₂₂H₂₆O₈。UV (MeOH) λ_max (log ε): 242 (2.15), 330 (2.33) nm。¹H NMR (CDCl₃, 600 MHz) 和¹³C NMR (CDCl₃, 150 MHz) 数据见表 1。

化合物2 淡黄色固体; HR-ESI-MS m/z 449.179 1 [M+H]⁺ (计算值为C₂₃H₂₉O₉⁺, 449.180 6), 分子式为C₂₃H₂₈O₉。UV (MeOH) λ_max (log ε): 250 (2.74), 348 (3.10) nm。¹H NMR (CDCl₃, 600 MHz) 和¹³C NMR (CDCl₃, 150 MHz) 数据见表 1。

化合物3 淡黄色固体; HR-ESI-MS m/z 287.091 2 [M+H]⁺ (计算值为C₁₆H₁₅O₅⁺, 287.091 4), 分子式为C₁₆H₁₄O₅。¹H NMR (CDCl₃, 400 MHz) δ_H 12.02 (1H, br s, 5-OH), 7.34 (2H, d, J = 8.5 Hz, H-2′, H-6′), 6.89 (2H, d, J = 8.5 Hz, H-3′, H-5′), 6.07 (1H, d, J = 2.3 Hz, H-8), 6.04 (1H, d, J = 2.3 Hz, H-6), 5.36 (1H, dd, J = 13.0, 3.0 Hz, H-2), 3.81 (3H, s, 7-OMe), 3.09 (1H, dd, J = 17.1, 13.0 Hz, H-3), 2.79 (1H, dd, J = 17.0, 3.0 Hz, H-3); ¹³C NMR (CDCl₃, 400 MHz) δ_C 79.1 (C-2), 43.4 (C-3), 196.2 (C-4), 164.3 (C-5), 95.3 (C-6), 168.1 (C-7), 94.4 (C-8), 163.0 (C-9), 103.3 (C-10), 130.8 (C-1′), 128.1 (C-2′), 115.8 (C-3′), 156.3 (C-4′), 115.8 (C-5′), 128.1 (C-6′), 55.8 (7-OMe)。以上波谱数据与文献^[14]报道数据基本一致, 故鉴定化合物3为樱花素。

化合物4 淡黄色固体; HR-ESI-MS m/z 345.096 4 [M+H]⁺ (计算值为C₁₈H₁₇O₇⁺, 345.096 9), 分子式为C₁₈H₁₆O₇。¹H NMR (CDCl₃, 600 MHz) δ_H 12.63 (1H, br s, 5-OH), 7.72 (1H, dd, J = 8.6, 2.0 Hz, H-6′), 7.69 (1H, d, J = 2.0 Hz, H-2′), 6.97 (1H, d, J = 8.6 Hz, H-5′), 6.45 (1H, d, J = 2.2 Hz, H-8), 6.35 (1H, d, J = 2.2 Hz, H-6), 5.73 (1H, s, 4′-OH), 3.99 (3H, s, 3′-OMe), 3.87 (3H, s, 3-OMe), 3.87 (3H, s, 7-OMe); ¹³C NMR (CDCl₃, 150 MHz) δ_C 155.8 (C-2), 139.3 (C-3), 179.0 (C-4), 162.1 (C-5), 98.0 (C-6), 165.6 (C-7), 92.3 (C-8), 156.9 (C-9), 106.2 (C-10), 123.8 (C-1′), 110.5 (C-2′), 145.6 (C-3′), 148.9 (C-4′), 114.5 (C-5′), 121.7 (C-6′), 60.3 (3-OMe), 56.0 (7-OMe), 56.2 (3′-OMe)。以上波谱数据与文献^[15]报道数据基本一致, 故鉴定化合物4为藿香黃酮醇。

化合物5 淡黄色固体; HR-ESI-MS m/z 359.112 1 [M+H]⁺ (计算值为C₁₉H₁₉O₇⁺, 359.112 5), 分子式为C₁₉H₁₈O₇。¹H NMR (CDCl₃, 600 MHz) δ_H 12.64 (1H, br s, 5-OH), 7.74 (1H, dd, J = 8.6, 2.0 Hz, H-6′), 7.69 (1H, d, J = 2.0 Hz, H-2′), 7.00 (1H, d, J = 8.6 Hz, H-5′), 6.45 (1H, d, J = 2.2 Hz, H-8), 6.37 (1H, d, J = 2.2 Hz, H-6), 3.98 (3H, s, 4′-OMe), 3.97 (3H, s, 3′-OMe), 3.88 (3H, s, 7-OMe), 3.87 (3H, s, 3-OMe); ¹³C NMR (CDCl₃, 150 MHz) δ_C 156.0 (C-2), 139.1 (C-3), 178.9 (C-4), 162.2 (C-5), 98.0 (C-6), 165.6 (C-7), 92.4 (C-8), 156.9 (C-9), 106.2 (C-10), 123.1 (C-1′), 111.4 (C-2′), 148.9 (C-3′), 151.5 (C-4′), 111.0 (C-5′), 122.3 (C-6′), 60.4 (3-OMe), 56.0 (7-OMe), 56.2 (3′-OMe), 56.2 (4′-OMe)。以上波谱数据与文献^[15]报道数据基本一致, 故鉴定化合物5为雷杜辛黄酮醇。

化合物6 淡黄色固体; HR-ESI-MS m/z 345.096 4 [M+H]⁺ (计算值为C₁₈H₁₇O₇⁺, 345.096 9), 分子式为C₁₈H₁₆O₇。¹H NMR (CDCl₃, 600 MHz) δ_H 12.60 (1H, br s, 5-OH), 8.04 (1H, d, J=8.4 Hz, H-2′), 8.04 (1H, d, J = 8.4 Hz, H-6′), 6.98 (1H, d, J = 8.4 Hz, H-5′), 6.98 (1H, d, J = 8.4 Hz, H-3′), 6.51 (1H, s, H-8), 5.70 (1H, s, 4′-OH), 3.96 (3H, s, 7-OMe), 3.93 (3H, s, 6-OMe), 3.86 (3H, s, 3-OMe); ¹³C NMR (CDCl₃, 150 MHz) δ_C 156.1 (C-2), 138.9 (C-3), 179.1 (C-4), 152.5 (C-5), 132.4 (C-6), 158.2 (C-7), 90.5 (C-8), 152.9 (C-9), 106.7 (C-10), 123.1 (C-1′), 130.6 (C-2′), 115.8 (C-3′), 158.9 (C-4′), 115.8 (C-5′), 130.6 (C-6′), 61.1 (3-OMe), 60.3 (6-OMe), 56.5 (7-OMe)。以上波谱数据与文献^[15]报道数据基本一致, 故鉴定化合物6为垂叶黄素。

化合物7 淡黄色固体; HR-ESI-MS m/z 375.106 9 [M+H]⁺ (计算值为C₁₉H₁₉O₈⁺, 375.107 4), 分子式为C₁₉H₁₈O₈。¹H NMR (CDCl₃, 600 MHz) δ_H 12.61 (1H, br s, 5-OH), 7.74 (1H, dd, J = 8.5, 2.2 Hz, H-6′), 7.70 (1H, d, J = 2.2 Hz, H-2′), 6.99 (1H, d, J = 8.5 Hz, H-5′), 6.52 (1H, s, H-8), 5.75 (1H, s, 5′-OH), 4.01 (3H, s, 6-OMe), 3.98 (3H, s, 3-OMe), 3.94 (3H, s, 7-OMe), 3.89 (3H, s, 4′-OMe); ¹³C NMR (CDCl₃, 150 MHz) δ_C 152.9 (C-2), 139.1 (C-3), 179.1 (C-4), 152.5 (C-5), 132.4 (C-6), 158.9 (C-7), 90.5 (C-8), 155.8 (C-9), 106.7 (C-10), 123.7 (C-1′), 110.5 (C-2′), 145.7 (C-3′), 148.9 (C-4′), 114.5 (C-5′), 121.7 (C-6′), 60.3 (3-OMe), 61.0 (6-OMe), 56.5 (7-OMe), 56.2 (4′-OMe)。以上波谱数据与文献^[16]报道数据基本一致, 故鉴定化合物7为蔓荆子黄素。

化合物8 淡黄色固体; HR-ESI-MS m/z 331.081 0 [M+H]⁺ (计算值为C₁₇H₁₅O₇⁺, 331.081 2), 分子式为C₁₇H₁₄O₇。¹H NMR (MeOD, 600 MHz) δ_H 7.64 (1H, d, J = 2.0 Hz, H-2′), 7.55 (1H, dd, J = 8.4; 2.0 Hz, H-6′), 6.90 (1H, d, J = 8.4 Hz, H-3′), 6.59 (1H, d, J = 2.1 Hz, H-8), 6.33 (1H, d, J = 2.1 Hz, H-6), 3.88 (3H, s, 3-OMe), 3.79 (3H, s, 7-OMe); ¹³C NMR (MeOD, 150 MHz) δ_C 158.3 (C-2), 139.7 (C-3), 180.1 (C-4), 162.8 (C-5), 98.9 (C-6), 167.3 (C-7), 93.1 (C-8), 158.4 (C-9), 106.7 (C-10), 122.8 (C-1′), 116.5 (C-2′), 146.5 (C-3′), 150.2 (C-4′), 116.4 (C-5′), 122.4 (C-6′), 60.5 (3-OMe), 56.5 (7-OMe)。以上波谱数据与文献^[17]报道数据基本一致, 故鉴定化合物8为3, 7-二甲氧基-5, 3′, 4′-三羟基黄酮。

化合物9 淡黄色固体; HR-ESI-MS m/z 361.091 3 [M+H]⁺ (计算值为C₁₈H₁₇O₈⁺, 361.091 8), 分子式为C₁₈H₁₆O₈。¹H NMR (MeOD, 600 MHz) δ_H 7.61~7.59 (2H, m, H-2′, H-6′), 7.02 (1H, d, J = 8.5 Hz, H-5′), 6.69 (1H, s, H-8), 3.96 (3H, s, 6-OMe), 3.93 (3H, s, 7-OMe), 3.78 (3H, s, 4′-OMe); ¹³C NMR (MeOD, 150 MHz) δ_C 151.7 (C-2), 139.8 (C-3), 180.2 (C-4), 151.1 (C-5), 131.1 (C-6), 157.8 (C-7), 91.6 (C-8), 155.9 (C-9), 107.2 (C-10), 124.3 (C-1′), 116.2 (C-2′), 146.7 (C-3′), 147.6 (C-4′), 112.3 (C-5′), 122.1 (C-6′), 60.6 (6-OMe), 57.0 (7-OMe), 56.4 (4′-OMe)。以上波谱数据与文献^[18]报道数据基本一致, 故鉴定化合物9为泽兰黄醇。

化合物10 无色油状物; [α]

$ {}_{\mathrm{D}}^{20} $

+4.60° (c 0.016 2, CH₃OH); HR-ESI-MS m/z 305.174 6 [M+H]⁺ (计算值为C₁₈H₂₅O₄⁺, 305.174 7), 分子式为C₁₈H₂₄O₄。¹H NMR (600 MHz, CDCl₃) 6.91~6.89 (1H, dd, J = 10.2, 3.8 Hz, H-4), 5.97 (1H, d, J = 10.2 Hz, H-3), 5.57 (1H, d, J = 10.1 Hz, H-1′), 5.14 (1H, d, J = 10.1 Hz, H-2′), 4.98 (1H, m, H-6′), 4.56 (1H, m, H-5), 3.17 (1H, d, J = 17.3 Hz, H-7), 2.79 (1H, br s, 5-OH), 2.30 (1H, d, J = 17.3 Hz, H-7), 2.32 (1H, m, H-6), 2.20 (1H, dd, J = 14.0; 5.1 Hz, H-6), 2.02 (2H, m, H-4′), 2.02 (2H, m, H-5′), 1.70 (3H, s, H-9′), 1.66 (3H, s, H-8′), 1.57 (3H, s, H-10′); ¹³C NMR (150 MHz, CDCl₃) δ_C 52.5 (C-1), 175.4 (C-2), 129.0 (C-3), 149.0 (C-4), 63.4 (C-5), 39.6 (C-6), 38.2 (C-7), 196.5 (C-8), 82.4 (C-1′), 118.5 (C-2′), 145.6 (C-3′), 39.8 (C-4′), 26.2 (C-5′), 123.4 (C-6′), 132.3 (C-7′), 25.8 (C-8′), 17.0 (C-9′), 17.8 (C-10′)。以上波谱数据与文献^[19]报道数据基本一致, 故鉴定化合物10为(+) -miliusol。

化合物11 无色油状物; [α]

$ {}_{\mathrm{D}}^{20} $

-1.97° (c 0.009 6, CH₃OH); HR-ESI-MS m/z 341.172 3 [M+Na]⁺ (计算值为C₁₉H₂₆NaO₄⁺, 341.172 3), 分子式为C₁₉H₂₆O₄。¹H NMR (600 MHz, CDCl₃) 7.24 (1H, m, H-4), 6.07 (1H, d, J = 9.6 Hz, H-3), 5.03 (1H, m, H-6′), 5.03 (1H, m, H-2′), 4.74 (1H, t, J = 5.4 Hz, H-5), 4.38 (1H, d, J = 9.5 Hz, H-1′), 3.63 (3H, s, 8-OMe), 2.77 (1H, d, J = 17.3 Hz, H-7), 2.74 (1H, d, J = 11.6 Hz, H-6), 2.22 (1H, d, J = 17.3 Hz, H-7), 2.09 (2H, m, H-5′), 2.06 (2H, m, H-4′), 2.04 (1H, m, H-6), 1.66 (3H, s, H-8′), 1.61 (3H, s, H-10′), 1.59 (3H, s, H-9′); ¹³C NMR (150 MHz, CDCl₃) δ_C 58.6 (C-1), 200.9 (C-2), 129.5 (C-3), 150.5 (C-4), 72.7 (C-5), 42.6 (C-6), 34.6 (C-7), 172.0 (C-8), 75.8 (C-1′), 120.2 (C-2′), 142.1 (C-3′), 40.0 (C-4′), 26.2 (C-5′), 123.7 (C-6′), 132.1 (C-7′), 25.7 (C-8′), 17.8 (C-9′), 16.7 (C-10′), 51.7 (8-OMe)。以上波谱数据与文献^[19]报道数据基本一致, 故鉴定化合物11为miliusane XIX。

化合物12 无色油状物; [α]

$ {}_{\mathrm{D}}^{20} $

+7.60° (c 0.000 1, CH₃OH); HR-ESI-MS m/z 337.200 9 [M+H]⁺ (计算值为C₁₉H₂₉O₅⁺, 337.201 0), 分子式为C₁₉H₂₈O₅。¹H NMR (600 MHz, CDCl₃) 6.90 (1H, d, J = 10.2 Hz, H-4), 6.04 (1H, d, J = 10.2 Hz, H-3), 5.05 (1H, m, H-6′), 5.02 (1H, m, H-2′), 4.79 (1H, s, H-5), 4.35 (1H, d, J = 6.0 Hz, H-1′), 3.78 (3H, s, 8-OMe), 2.40 (1H, m, H-6), 2.36 (2H, m, H-7), 2.30 (1H, m, H-7), 2.06 (2H, m, H-5′), 2.00 (2H, m, H-4′), 1.95 (1H, m, H-6), 1.68 (3H, s, H-8′), 1.62 (3H, s, H-10′), 1.60 (3H, s, H-9′); ¹³C NMR (150 MHz, CDCl₃) δ_C 53.6 (C-1), 199.7 (C-2), 129.4 (C-3), 151.1 (C-4), 64.9 (C-5), 38.0 (C-6), 30.5 (C-7), 173.5 (C-8), 73.8 (C-1′), 118.5 (C-2′), 139.4 (C-3′), 40.1 (C-4′), 26.6 (C-5′), 124.2 (C-6′), 131.7 (C-7′), 25.9 (C-8′), 17.9 (C-9′), 16.4 (C-10′), 52.7 (8-OMe)。以上波谱数据与文献^[19]报道数据基本一致, 故鉴定化合物12为miliusane XVIII。

化合物13淡黄色油状物; HR-ESI-MS m/z 357.241 8 [M+H]⁺ (计算值为C₂₃H₃₃O₃⁺, 357.242 4), 分子式为C₂₃H₃₂O₃。¹H NMR (CDCl₃, 400 MHz) δ_H 6.03 (1H, dt, J = 10.8, 7.4 Hz, H-16), 5.81 (1H, m, H-19), 5.49 (1H, m, H-15), 5.02 (2H, m, H-20), 4.59 (1H, m, H-22), 3.86 (1H, dd, J = 12.3, 3.0 Hz, H-23), 3.65 (1H, dd, J = 12.3, 3.0 Hz, H-23), 2.70 (1H, m, H-2), 2.44 (2H, m, H-17), 2.32 (1H, m, H-21), 2.17 (2H, m, H-18), 2.00 (1H, m, H-21), 1.83 (1H, m, H-3), 1.54 (2H, m, H-9), 1.45 (1H, m, H-3), 1.30 (10H, m, H-4-8); ¹³C NMR (CDCl₃, 100 MHz) δ_C 179.8 (C-1), 39.7 (C-2), 31.4 (C-3), 29.4~27.3 (C-4-8), 28.3 (C-9), 19.7 (C-10), 85.2 (C-11), 65.3 (C-12), 72.1 (C-13), 77.4 (C-14), 108.8 (C-15), 146.8 (C-16), 30.0 (C-17), 33.0 (C-18), 137.8 (C-19), 115.3 (C-20), 29.7 (C-21), 78.6 (C-22), 64.8 (C-23)。以上波谱数据与文献^[20]报道数据基本一致, 故鉴定化合物13为miliusolide。

化合物14 淡黄色油状物; HR-ESI-MS m/z 359.257 4 [M+H]⁺ (计算值为C₂₃H₃₅O₃⁺, 359.258 1), 分子式为C₂₃H₃₄O₃。¹H NMR (CDCl₃, 600 MHz) δ_H 6.03 (1H, dt, J = 10.8, 7.5Hz, H-12), 5.46 (1H, d, J = 10.8 Hz, H-15), 4.59 (1H, m, H-22), 3.86 (1H, dd, J = 12.4, 3.8 Hz, H-23), 3.65 (1H, dd, J = 12.4, 3.8 Hz, H-23), 2.69 (1H, m, H-2), 2.32 (2H, t, J = 6.9 Hz, H-10), 2.32 (2H, m, H-17), 2.27 (1H, m, H-21), 2.01 (1H, m, H-21), 1.83 (1H, m, H-3), 1.54 (2H, m, H-9), 1.47 (1H, m, H-3), 1.38 (2H, m, H-18), 1.35 (2H, m, H-19), 1.29~1.40 (10H, m, H-4-8), 0.91 (3H, t, J = 7.0 Hz, H-20); ¹³C NMR (CDCl₃, 150 MHz) δ_C 179.8 (C-1), 39.7 (C-2), 31.4 (C-3), 29.4~27.3 (C-4-8), 28.4 (C-9), 19.7 (C-10), 84.9 (C-11), 65.3 (C-12), 72.3 (C-13), 78.2 (C-14), 108.2 (C-15), 148.0 (C-16), 30.5 (C-17), 31.1 (C-18), 22.4 (C-19), 14.1 (C-20), 29.7 (C-21), 78.6 (C-22), 64.8 (C-23)。以上波谱数据与文献^[20]报道数据基本一致, 故鉴定化合物14为19, 20-dihydromiliusolide。

化合物15 淡黄色油状物; HR-ESI-MS m/z 409.271 5 [M+Na]⁺ (计算值为C₂₅H₃₈NaO₃⁺, 409.271 3), 分子式为C₂₅H₃₈O₃。¹H NMR (CDCl₃, 600 MHz) δ_H 6.03 (1H, dt, J = 10.8, 7.5 Hz, H-18), 5.46 (1H, d, J = 10.8 Hz, H-17), 4.60 (1H, m, H-24), 3.86 (1H, dd, J = 12.4, 3.1 Hz, H-25), 3.65 (1H, dd, J = 12.4, 3.1 Hz, H-25), 2.69 (1H, m, H-2), 2.32 (2H, t, J = 7.3 Hz, H-12), 2.32 (2H, m, H-19), 2.28 (1H, m, H-23), 2.00 (1H, m, H-23), 1.83 (1H, m, H-3), 1.54 (2H, m, H-11), 1.46 (1H, m, H-3), 1.38 (2H, m, H-20), 1.33 (2H, m, H-21), 1.27~1.38 (14H, m, H-4-10), 0.91 (3H, t, J = 7.1 Hz, H-22); ¹³C NMR (CDCl₃, 150 MHz) δ_C 179.8 (C-1), 39.7 (C-2), 31.4 (C-3), 29.7~27.4 (C-4-10), 28.4 (C-11), 19.8 (C-12), 85.0 (C-13), 65.3 (C-14), 72.3 (C-15), 78.2 (C-16), 108.2 (C-17), 148.0 (C-18), 30.6 (C-19), 31.1 (C-20), 22.4 (C-21), 14.1 (C-22), 29.4 (C-23), 78.6 (C-24), 64.8 (C-25)。以上波谱数据与文献^[21]报道数据基本一致, 故鉴定化合物15为debilisone C。

化合物16 淡黄色油状物; HR-ESI-MS m/z 391.320 3 [M+H]⁺ (计算值为C₂₅H₄₃O₃⁺, 391.320 7), 分子式为C₂₅H₄₂O₃。¹H NMR (CDCl₃, 400 MHz) δ_H 5.80 (1H, ddt, J = 16.9, 10.1, 6.7 Hz, H-21), 4.98 (1H, dd, J = 17.3, 2.0 Hz, H-22), 4.92 (1H, d, J = 9.9 Hz, H-22), 4.58 (1H, m, H-24), 3.86 (1H, dd, J = 12.4, 3.0 Hz, H-25), 3.64 (1H, dd, J = 12.4, 4.6 Hz, H-25), 2.69 (1H, m, H-2), 2.29 (1H, m, H-23), 2.13 (2H, t, J = 6.7 Hz, H-12), 2.13 (2H, t, J = 6.7 Hz, H-15), 2.03 (2H, m, H-20), 1.99 (1H, m, H-23), 1.82 (1H, m, H-3), 1.46~1.26 (14H, m, H-5-11), 1.46~1.26 (8H, m, H-16-19), 1.45 (1H, m, H-3), 1.37 (2H, m, H-4); ¹³C NMR (CDCl₃, 100 MHz) δ_C 180.0 (C-1), 39.7 (C-2), 31.4 (C-3), 29.6~28.8 (C-4-11), 18.8 (C-12), 80.4 (C-13), 80.3 (C-14), 18.8 (C-15), 29.6~28.8 (C-16-19), 33.9 (C-20), 139.2 (C-21), 114.3 (C-22), 29.7 (C-23), 78.7 (C-24), 64.7 (C-25)。以上波谱数据与文献^[22]报道数据基本一致, 故鉴定化合物16为goniothalamusin。

化合物17 棕色胶状物; [α]

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-21.0° (c 0.009 3, CH₃OH), HR-ESI-MS m/z 342.133 1 [M+H]⁺ (计算值为C₁₉H₂₀NO₅⁺, 342.133 6), 分子式为C₁₉H₁₉NO₅。¹H NMR (MeOD, 600 MHz) δ_H 6.98 (1H, m, H-11), 6.94 (1H, m, H-12), 6.84 (1H, m, H-4), 6.64 (1H, m, H-1), 4.84 (1H, m, H-6), 4.73 (1H, m, H-13a), 3.85 (3H, s, 3-OMe), 3.83 (3H, s, 9-OMe), 3.15 (1H, m, H-13), 2.91 (1H, m, H-6), 2.80 (1H, m, H-5), 2.72 (1H, m, H-5), 2.72 (1H, m, H-13); ¹³C NMR (MeOD, 150 MHz) δ_C 115.9 (C-1), 128.8 (C-1b), 146.6 (C-2), 148.3 (C-3), 110.7 (C-4), 128.0 (C-4a), 30.0 (C-5), 39.8 (C-6), 165.0 (C-8), 123.1 (C-8a), 149.4 (C-9), 151.4 (C-10), 120.8 (C-11), 124.0 (C-12), 132.0 (C-12a), 39.7 (C-13), 56.6 (C-13a), 56.6 (3-OMe), 62.0 (9-OMe)。以上波谱数据与文献^[23]报道数据基本一致, 故鉴定化合物17为2, 10-dihydroxy-3, 9-dimethoxy-8-oxo-protoberberine。

化合物18 棕色胶状物; HR-ESI-MS m/z 328.117 6 [M+H]⁺ (计算值为C₁₈H₁₈NO₅⁺, 328.117 9), 分子式为C₁₈H₁₇NO₅。¹H NMR (CDCl₃, 400 MHz) δ_H 12.75 (1H, s, 9-OH), 6.99 (1H, d, J = 8.0 Hz, H-11), 6.80 (1H, s, H-1), 6.67 (1H, s, H-4), 6.61 (1H, dd, J = 8.0, 1.2 Hz, H-12), 5.58 (s, 2H, 10-OH, 2-OH), 4.83 (1H, m, H-6), 4.83 (1H, m, H-13a), 3.91 (3H, s, 3-OMe), 3.16 (1H, dd, J = 15.6, 3.9 Hz, H-13), 2.99 (1H, m, H-14), 2.94 (1H, m, H-5), 2.85 (1H, m, H-13), 2.79 (1H, m, H-5); ¹³C NMR (CDCl₃, 100 MHz) δ_C 111.3 (C-1), 128.4 (C-1b), 145.8 (C-2), 144.8 (C-3), 111.7 (C-4), 126.2 (C-4a), 29.1 (C-5), 38.6 (C-6), 168.7 (C-8), 110.8 (C-8a), 148.6 (C-9), 144.0 (C-10), 118.3 (C-11), 117.0 (C-12), 128.1 (C-12a), 36.8 (C-13), 55.6 (C-13a), 56.2 (3-OMe)。以上波谱数据与文献^[24]报道数据基本一致, 故鉴定化合物18为consanguine B。

化合物19 无色油状物; HR-ESI-MS m/z 427.356 8 [M+H]⁺ (calculated for C₂₉H₄₇O₂⁺: 427.357 1), 分子式为C₂₉H₄₆O₂。¹H NMR (CDCl₃, 400 MHz) δ_H 5.81 (1H, s, H-4), 5.14 (1H, dd, J = 15.2, 8.5 Hz, H-22), 5.03 (1H, dd, J = 15.2, 8.5 Hz, H-23), 4.34 (1H, m, H-6), 2.52 (1H, m, H-2), 2.41 (1H, m, H-2), 2.07 (1H, m, H-20), 2.06 (1H, m, H-1), 2.05 (1H, m, H-12), 2.02 (1H, m, H-7), 1.98 (1H, m, H-8), 1.89 (1H, m, H-28), 1.75 (2H, m, H-1, H-16), 1.65 (1H, m, H-15), 1.55 (2H, m, H-25, H-24), 1.53 (2H, m, H-11), 1.44 (1H, m, H-28), 1.38 (3H, s, H-19), 1.31 (1H, m, H-16), 1.25 (1H, m, H-7), 1.21 (1H, m, H-12), 1.19 (1H, m, H-17), 1.18 (1H, m, H-15), 1.06 (1H, m, H-14), 1.02 (3H, d, J = 6.6 Hz, H-21), 0.93 (1H, m, H-9), 0.85 (3H, d, J = 6.1 Hz, H-26), 0.81 (3H, d, J = 7.2 Hz, H-29), 0.80 (3H, d, J = 5.9 Hz, H-27), 0.76 (3H, s, H-18); ¹³C NMR (CDCl₃, 100 MHz) δ_C 37.2 (C-1), 34.4 (C-2), 200.6 (C-3), 126.5 (C-4), 168.6 (C-5), 73.4 (C-6), 38.7 (C-7), 29.9 (C-8), 53.8 (C-9), 38.1 (C-10), 21.1 (C-11), 39.6 (C-12), 42.5 (C-13), 56.1 (C-14), 24.4 (C-15), 29.0 (C-16), 56.1 (C-17), 12.3 (C-18), 19.6 (C-19), 40.6 (C-20), 21.3 (C-21), 138.3 (C-22), 129.6 (C-23), 51.4 (C-24), 32.0 (C-25), 21.2 (C-26), 19.1 (C-27), 25.5 (C-28), 12.4 (C-29)。以上波谱数据与文献^{[25, 26]}报道数据基本一致, 故鉴定化合物19为6-羟基豆甾-4, 22-二烯-3-酮。

化合物20 无色油状物; HR-ESI-MS m/z 429.372 6 [M+H]⁺ (计算值为C₂₉H₄₉O₂⁺, 429.372 7), 分子式为C₂₉H₄₈O₂。¹H NMR (CDCl₃, 400 MHz) δ_H 5.80 (1H, s, H-4), 4.34 (1H, s, H-6), 2.51 (1H, m, H-2), 2.40 (1H, m, H-2), 2.07 (1H, m, H-12), 2.05 (1H, m, H-1), 2.02 (1H, m, H-7), 1.97 (1H, m, H-8), 1.88 (1H, m, H-16), 1.73 (1H, m, H-1), 1.68 (1H, m, H-25), 1.63 (1H, m, H-15), 1.52 (2H, m, H-11), 1.39 (1H, m, H-20), 1.37 (3H, s, H-19), 1.34 (1H, m, H-22), 1.32 (1H, m, H-16), 1.27 (2H, m, H-28), 1.26 (1H, m, H-7), 1.18 (2H, m, H-23), 1.17 (2H, m, H-15, H-12), 1.03 (3H, m, H-22, H-17, H-14), 0.95 (1H, m, H-24), 0.92 (1H, m, H-9), 0.92 (3H, d, J = 6.6 Hz, H-21), 0.85 (3H, d, J = 7.7 Hz, H-29), 0.83 (3H, d, J = 7.2 Hz, H-27), 0.81 (3H, d, J = 6.8 Hz, H-26), 0.74 (3H, s, H-18); ¹³C NMR (CDCl₃, 100 MHz) δ_C 37.2 (C-1), 34.4 (C-2), 200.7 (C-3), 126.4 (C-4), 168.8 (C-5), 73.4 (C-6), 38.7 (C-7), 29.9 (C-8), 53.8 (C-9), 38.1 (C-10), 21.1 (C-11), 39.7 (C-12), 42.6 (C-13), 56.0 (C-14), 24.3 (C-15), 28.3 (C-16), 56.2 (C-17), 12.1 (C-18), 19.6 (C-19), 36.3 (C-20), 18.9 (C-21), 34.0 (C-22), 26.2 (C-23), 46.0 (C-24), 29.3 (C-25), 20.0 (C-26), 19.2 (C-27), 23.2 (C-28), 12.1 (C-29)。以上波谱数据与文献^[26]报道数据基本一致, 故鉴定化合物20为6-羟基豆甾-4-烯-3-酮。

3 抗肿瘤活性实验

将测试化合物分别用二甲基亚砜(DMSO) 溶解, 配制成1.0×10² mmol·L^-1浓度的储备药液, 分别用含10%胎牛血清的DMEM和RPMI-1640培养液将储备药液稀释成具有8个浓度梯度(100、50、25、12.5、6.25、3.125、1.562 5、0.781 25 μmol·L^-1) 的待测药液。取对数生长期的人乳腺癌细胞株MDA-MB-231和BT-549, 将细胞按5 000个/孔的密度接种于96孔板中, 每孔100 μL细胞悬液, 孵育24 h后, 分别加入100 μL不同浓度待测药液作用24 h, 然后每孔加入5 mg·mL^-1的MTT溶液20 μL, 放入培养箱中孵育3~4 h。待反应充分后弃去上清液, 每孔加入150 μL DMSO, 在96孔板振荡器上低速振荡5~10 min左右使结晶紫充分溶解, 用酶标仪测定吸光度值, 测定波长为490 nm。分析计算细胞活性: 细胞存活率= (实验孔-空白孔) / (对照孔-空白孔) ×100%。每个数据平行测定3次, 取其平均值。

作者贡献: 刘志莹是本文的第一作者, 负责云南野独活化学成分的分离纯化及结构解析工作; 葛格负责活性测试工作; 刘庭亮和郑文琳对提取分离和结构鉴定提供了一定帮助; 郭亚萍负责论文的审阅和提出写作的建议; 王书云和吴炎负责实验的整体设计与指导工作及论文的审阅。

利益冲突: 所有作者均声明不存在利益冲突。

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2025年第60卷第3期

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doi: 10.16438/j.0513-4870.2024-0772

接收时间：2024-08-12
首发时间：2025-11-06
出版时间：2025-03-12

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国家自然科学基金资助项目(82004061)

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1.河南大学药学院, 河南开封 475004

2.深圳技术大学药学院, 广东深圳 518118

通讯作者:

^*王书云, E-mail: wangshuyun0426@163.com

吴炎, E-mail: wuyan1219@yeah.net

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https://castjournals.cast.org.cn/joweb/yxxb/CN/10.16438/j.0513-4870.2024-0772

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2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

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No.	1		2
No.	δ_H	δ_C	δ_H	δ_C
1		127.6		130.1
2	7.07 (1H, d, J = 2.2 Hz)	110.0	6.76 (1H, s)	105.8
3		149.4		153.6
3-OMe	3.91 (3H, s)	56.1	3.88 (3H, s)	56.4
4		151.6		140.6
4-OMe	3.91 (3H, s)	56.1	3.88 (3H, s)	61.2
5	6.85 (1H, d, J = 8.2 Hz)	111.1		153.6
5-OMe			3.88 (3H, s)	56.4
6	7.10 (1H, dd, J = 8.2, 2.2 Hz)	123.6	6.76 (1H, s)	105.8
1′		124.3		124.2
2′		146.2		146.3
2′-OMe	3.80 (3H, s)	62.2	3.81 (3H, s)	62.2
3′		143.2		143.2
3′-OMe	3.91 (3H, s)	61.4	3.91 (3H, s)	61.5
4′		148.8		148.9
4′-OMe	3.99 (3H, s)	61.6	3.99 (3H, s)	61.6
5′		143.2		143.2
5′-OMe	3.91 (3H, s)	61.4	3.91 (3H, s)	61.5
6′		146.2		146.3
6′-OMe	3.80 (3H, s)	62.2	3.81 (3H, s)	62.2
α	6.88 (1H, d, J = 16.0 Hz)	126.8	6.90 (1H, d, J = 15.9 Hz)	128.1
β	7.27 (1H, d, J = 16.0 Hz)	146.3	7.25 (1H, d, J = 15.9 Hz)	146.0
C=O		193.7		193.5