CN116478029B - 木麻黄醇e及其制备方法和在抗炎药物中的应用 - Google Patents
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Abstract
本发明公开了木麻黄醇E及其制备方法和在抗炎药物中的应用。所述的木麻黄醇E其结构如式(1)所示。本发明发现了一种新的、具有良好抗炎活性的大环二苯庚烷木麻黄醇E，它在抗炎药物开发方面具有广泛的应用前景。。
Description
技术领域
本发明属于天然药物领域，具体涉及一种新的大环二苯庚烷-木麻黄醇E及其制备方法和在制备抗炎药物中的应用。
背景技术
炎症介质是指参与和介导炎症反应的化学因子，如可溶性气体NO，细胞因子IL-1、IL-6、和TNF-α，前列腺素等，可导致发热疼痛和组织损伤，这些炎症介质可维持和发展炎症反应。因此，一些能够下调这些炎症介质的药物可用于临床抗炎止痛，如塞来昔布。然而现有的抗炎药物对有些病人无效，长期使用这些抗炎药物会产生不良反应。因此，有必要寻找和开发新型的抗炎药物。
木麻黄(Casuarina equisetifolia L.)系木麻黄科木麻黄属常绿乔木，原产于澳大利亚及太平洋岛屿，现在我国广东、广西、福建、台湾和浙江广泛种植，常用于沿海防风林，目前面积逾30万hm2。根据《中国中药资源大典》和《中华本草》记载，木麻黄树皮、枝叶、种子可入药，主要功效为止泻止痢。其中，关于木麻黄嫩枝、叶、树皮、凋落物的化学成分及现代药理活性研究较多，但对木麻黄果实中活性化学成分的研究基本没有报道。
迄今，未见木麻黄中具抗炎活性的大环二苯庚烷成分的研究报道。
发明内容
本发明的第一个目的是提供一种来源于木麻黄(Casuarina equisetifolia L.)果实的大环二苯庚烷-木麻黄醇E、或其药学上可接受的酯化衍生物。
本发明的木麻黄醇E，其结构如式(1)所示：
本发明的第二个目的在于提供式(1)所示的木麻黄醇E的制备方法，其特征在于，木麻黄醇E是从木麻黄(Casuarina equisetifolia L.)果实中制备分离得到的。
具体材料可以是干品或鲜品。
具体步骤优选为：
a、制备总浸膏：将木麻黄的果实粉碎后用有机溶剂-水的混合溶液浸提，提取液浓缩除去有机溶剂后得到混悬液，混悬液依次用石油醚、乙酸乙酯、正丁醇萃取，乙酸乙酯萃取液经减压浓缩后得到总浸膏；
b、分离纯化：总浸膏经正相硅胶柱层析，以二氯甲烷-甲醇从体积比100:0→0:100洗脱，收集二氯甲烷-甲醇体积比95:5洗脱的馏分为E13，馏分E13进一步经正相硅胶柱层析，以二氯甲烷-甲醇体积比从200:1→0:100洗脱，收集二氯甲烷-甲醇体积比50:1洗脱的馏分为E13-44，取馏分E13-44经凝胶柱层析，以二氯甲烷-甲醇体积比1:1洗脱，所得馏分经纯化得到大环二苯庚烷。
优选，所述的馏分经纯化是经岛津LC-6AD半制备高效液相色谱，色谱柱为Cosmosil5C18-MS-II column 5μm,10mm×250mm、流动相为乙腈-水体积比30:70、流速为2mL/min，收集保留时间为22min的馏分，获得木麻黄醇E。
所述的有机溶剂-水的混合溶液优选为体积分数95％的乙醇水溶液。
本发明的第三个目的是提供木麻黄果实在制备如式(1)所示的木麻黄醇E中的应用。
本发明式(1)所示的木麻黄醇E能够明显抑制LPS诱导下小鼠巨噬细胞中一氧化氮(NO)的产生。半数抑制浓度IC50为10.91±2.63μM，阳性对照药物氨基胍盐酸盐的IC50为6.72±1.20μM。说明木麻黄醇E具有良好的抗炎活性，是新的抗炎活性物质，有望作为先导化合物开发出新型的抗炎药物。
因此，本发明的第四个目的在于提供式(1)所示的木麻黄醇E、或其药学上可接受的酯化衍生物在制备抗炎药物中的应用。
本发明的第五个目的是提供一种抗炎药物，其特征在于，含有式(1)所示的木麻黄醇E、或其药学上可接受的酯化衍生物作为活性成分。
本发明发现了一种新的、具有良好抗炎活性的木麻黄醇E，它在抗炎药物开发方面具有广泛的应用前景。
附图说明
图1是化合物木麻黄醇E的1H NMR谱；
图2是化合物木麻黄醇E的13C NMR谱；
图3是化合物木麻黄醇E的HSQC谱；
图4是化合物木麻黄醇E的1H-1H COSY谱；
图5是化合物木麻黄醇E的HMBC谱；
图6是化合物木麻黄醇E的NOESY谱；
图7是化合物木麻黄醇E的HR-ESIMS谱；
图8是化合物木麻黄醇E的CD谱；
图9是化合物木麻黄醇E的UV谱；
具体实施方式
以下实施例是对本发明的进一步说明，不是对本发明的限制。
实施例1：本发明式(1)所示木麻黄醇E的制备
1.1仪器与试剂
供提取用植物材料木麻黄(Casuarina equisetifolia L.)的果实样品于2021年7-9月从华南植物园中木麻黄树下收集。柱层析硅胶(80～100目和200～300目)、薄层硅胶层析板均为烟台江友硅胶开发有限公司生产。葡聚糖凝胶Sephadex LH-20由瑞典AmershamBioscien ces公司生产。NMR谱用Bruker AVIII 500M型超导核磁共振仪测定，以氘代二甲基亚砜残留溶剂峰作标定，氘代二甲基亚砜为美国剑桥公司(CIL)生产。旋光用PerkinElmer 341plus旋光仪测定。CD谱用Jasco J-810圆二色谱仪测定。质谱采用API 2000LC/MS/MS质谱仪测定。
1.2木麻黄醇E的制备和结构鉴定
成熟晒干的木麻黄果实，经过粉碎，95％乙醇浸泡提取，提取液浓缩得到浸膏(641g)。浸膏用水溶，依次经过石油醚、乙酸乙酯、正丁醇萃取。萃取液浓缩，所得乙酸乙酯部位(60.3g)经正相硅胶柱层析，以二氯甲烷-甲醇(体积比从100:0→0:100)洗脱，收集二氯甲烷-甲醇体积比95:5洗脱的馏分为E13。馏分E13进一步经正相硅胶柱层析，以二氯甲烷-甲醇体积比从200:1→0:100洗脱，收集二氯甲烷-甲醇体积比50:1洗脱的馏分为E13-44；馏分E13-44经凝胶柱层析，以二氯甲烷-甲醇体积比1:1洗脱，所得馏分经岛津LC-6AD半制备高效液相色谱，色谱柱为Cosmosil 5C18-MS-II column(5μm,10mm×250mm)、流动相为乙腈-水体积比30:70、流速为2mL/min，收集保留时间为22min的馏分，获得木麻黄醇E。经结构鉴定为式(1)所示的新化合物。
式(1)所示的木麻黄醇E结构鉴定数据如下：[α]D 20＝+15.5(c 0.2,CH3OH)；ESI-MS:m/z 341[M-H]-；高分辨质谱m/z:343.1180[M+H]+，计算值：343.1176，分子式为C19H18O6。
图1是化合物木麻黄醇E的1H NMR谱；图2是化合物木麻黄醇E的13C NMR谱；图3是化合物木麻黄醇E的HSQC谱；图4是化合物木麻黄醇E的1H-1H COSY谱；图5是化合物木麻黄醇E的HMBC谱；图6是化合物木麻黄醇E的NOESY谱；图7是化合物木麻黄醇E的HR-ESIMS谱；图8是化合物木麻黄醇E的CD谱；图9是化合物木麻黄醇E的UV谱；
通过NOESY谱图确定结构中的相对构型为2R*,6S*。通过实验CD谱图与量子化学计算的CD谱图比较(图8)，确定绝对构型为2R,6S。
表1.木麻黄醇E的NMR数据(测试溶剂为氘代二甲基亚砜)
实施例2：式(1)所示的木麻黄醇E的体外抗炎活性2.1实验材料
小鼠巨噬细胞RAW264.7，DMEM培养基，NO测定试剂盒。
2.2木麻黄醇E对小鼠巨噬细胞毒性评价
将细胞以1.7×105个/mL的细胞浓度，每孔100μL铺于96孔板中(四周的孔加入100μL的PBS，避免边际效应)，培养箱中孵育24h后，将96孔板中的培养基吸出。加入溶解了不同浓度木麻黄醇E的DMEM培养基，以未加样品的细胞孔作为对照组，每组设置3个平行孔。将96孔板重新放置于培养箱中继续培养24h后，采用cck-8测定细胞活力。按下列公式求出细胞存活率，结果为木麻黄醇E在10μM和50μM浓度时细胞存活率分别为99±6％和95±5％，表明木麻黄醇E在50μM浓度及以下时对小鼠巨噬细胞没有毒性。
计算公式：细胞存活率(％)＝样品组吸光度×100/对照组吸光度
2.3体外抗炎活性评价
将细胞按1.7×104个/孔接种到96孔板中，孵育12h后，吸去培养基，加不含血清的培养基饥饿处理12h。弃去培养基后，分别加入LPS(200ng/mL)以及6.25、12.5、25和50μM四个梯度浓度的木麻黄醇E共孵育20h后，收集上清液，测定其NO浓度，其中不加LPS的培养基作为空白对照组，仅加LPS的培养基作为模型对照组。NO浓度测定采用Griess法，在540nm波长下测定吸光度。按如下公式计算抑制率，再由统计软件计算半数抑制浓度(IC50)。
计算公式：抑制率(％)＝(ODL-OD样)×100/(ODL-OD空)
ODL：LPS模型对照组的吸光度；OD样：样品组的吸光度；OD空：空白对照组的吸光度。
2.4抗炎活性结果
结果见表2，其中IC50用Mean±SD表示。
表2.木麻黄醇E对LPS诱导下小鼠巨噬细胞NO生成抑制IC50
Claims (6)
1.木麻黄醇E，其结构如式（1）所示：
式（1）。
2.一种权利要求1所述的木麻黄醇E的制备方法，其特征在于，具体步骤为：
a、制备总浸膏：将木麻黄的果实粉碎后用有机溶剂-水的混合溶液浸提，提取液浓缩除去有机溶剂后得到混悬液，混悬液依次用石油醚、乙酸乙酯、正丁醇萃取，乙酸乙酯萃取液经减压浓缩后得到总浸膏；
b、分离纯化：总浸膏经正相硅胶柱层析，以二氯甲烷-甲醇从体积比100:0→0:100洗脱，收集二氯甲烷-甲醇体积比95:5洗脱的馏分为E13，馏分E13进一步经正相硅胶柱层析，以二氯甲烷-甲醇体积比从200:1→0:100洗脱，收集二氯甲烷-甲醇体积比50:1洗脱的馏分为E13-44，取馏分E13-44经凝胶柱层析，以二氯甲烷-甲醇体积比1:1洗脱，所得馏分经纯化得到木麻黄醇E。
3.根据权利要求2所述的制备方法，其特征在于，所述的所得馏分经纯化得到木麻黄醇E是：所得馏分经岛津LC-6AD半制备高效液相色谱，色谱柱为Cosmosil 5C18-MS-II column5 µm, 10 mm×250 mm、流动相为乙腈-水体积比30:70、流速为2 mL/min，收集保留时间为22 min的馏分，获得木麻黄醇E。
4.根据权利要求2所述的制备方法，其特征在于，所述的有机溶剂-水的混合溶液为体积分数95%的乙醇水溶液。
5.权利要求1所述的木麻黄醇E在制备抗炎药物中的应用。
6.一种抗炎药物，其特征在于，含有权利要求1所述的木麻黄醇E作为活性成分。
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20140100117A (ko) * | 2013-02-05 | 2014-08-14 | 주식회사 엘컴사이언스 | 오리나무 추출물 또는 그로부터 분리된 화합물을 포함하는 지방분화 억제용 조성물 |
| CN115260022A (zh) * | 2022-08-16 | 2022-11-01 | 中国科学院华南植物园 | 一种大环二苯庚烷及其制备方法和在制备抗流感病毒药物中的应用 |
-
2023
- 2023-04-14 CN CN202310405409.0A patent/CN116478029B/zh active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20140100117A (ko) * | 2013-02-05 | 2014-08-14 | 주식회사 엘컴사이언스 | 오리나무 추출물 또는 그로부터 분리된 화합물을 포함하는 지방분화 억제용 조성물 |
| CN115260022A (zh) * | 2022-08-16 | 2022-11-01 | 中国科学院华南植物园 | 一种大环二苯庚烷及其制备方法和在制备抗流感病毒药物中的应用 |
Non-Patent Citations (9)
| Title |
|---|
| Anti-inflammatory activities for the extracts and carpinontriols from branches of Carpinus turczaninowii;Ko, Ha Na等;International Journal of Pharmacology;20131231;第9卷(第2期);157-163 * |
| Extractives of the genus Corylus. II. Novel diarylheptanoids from the wood of Corylus sieboldiana Blume. (1);Watanabe, Naoki等;Mokuzai Gakkaishi;19941231;第40卷(第2期);199-203 * |
| MASAYASU NOMURA等.BIARYLHEPTANOIDS AND OTHER CONSTITUENTS FROM WOOD OF ALNUS JAPONICA.Phytochemistry.1981,第20卷(第5期),第1097-1104页. * |
| MASAYASU NOMURA等.Three New Cyclized C9-C1-C9 Compounds from Alnus japonica Steud..J.C.S. CHEM. COMM..1974,第65-66页. * |
| Milena Masullo等.Giffonins, Antioxidant Diarylheptanoids from Corylus avellana, and Their Ability to Prevent Oxidative Changes in Human Plasma Proteins.Journal of Natural Products.2021,第84卷第646-653页. * |
| Xiaoyu Xu等.Discovery of Cyclic Diarylheptanoids as Inhibitors against Influenza A Virus from the Roots of Casuarina equisetifolia.Journal of Natural Products.2022,第85卷第2142-2148页. * |
| 大环二芳基庚烷类化合物的波普特征;张毅 等;波谱学杂志;20101231;第27卷(第4期);669-676 * |
| 天然产物中环状二芳基庚烷类化合物的研究进展;张建斌 等;中草药;20080812;第39卷(第8期);1263-1268 * |
| 天然大环二芳基庚烷类化合物的研究概况;刘红星 等;天然产物研究与开发;20080215;第20卷(第1期);172-179 * |
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