CN111544458A - 麦吊云杉提取物及其制备方法和在制药中的用途 - Google Patents

麦吊云杉提取物及其制备方法和在制药中的用途 Download PDF

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CN111544458A
CN111544458A CN201910111443.0A CN201910111443A CN111544458A CN 111544458 A CN111544458 A CN 111544458A CN 201910111443 A CN201910111443 A CN 201910111443A CN 111544458 A CN111544458 A CN 111544458A
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胡金锋
江维
熊娟
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Abstract

本发明涉及医药与保健品技术领域，涉及麦吊云杉提取物及其制备方法和在制药中的用途，本发明公开了麦吊云杉提取物包括粗提取物和单一成分的制备方法，以及作为腺苷三磷酸‑柠檬酸裂解酶[adenosine triphosphate(ATP)‑citrate lyase，ACL]抑制剂的药物用途。本发明从松科(Pinaceae)云杉属植物麦吊云杉[Picea brachytyla(Franch.)Pritz.]枝叶中提取和制备粗提取物及其萜类单体化合物，经实验证实，所述粗提物和萜类单体化合物均能显著抑制ACL活性，可作为ACL抑制剂或先导化合物用于制备糖脂代谢紊乱相关疾病的治疗药物或保健品；所述糖脂代谢相关代谢疾病包括高脂血症、动脉粥样硬化、非酒精性脂肪肝、II型糖尿病及肥胖症。

Description

麦吊云杉提取物及其制备方法和在制药中的用途
技术领域
本发明涉及医药与保健品技术领域，涉及麦吊云杉提取物及其制备方法和在制药中的用途，尤其是作为ACL抑制剂在制备治疗ATP-柠檬酸裂解酶介导疾病药物中的用途。
背景技术
现有技术公开了腺苷三磷酸-柠檬酸裂解酶[adenosine triphosphate(ATP)-citrate lyase，ACL]是肝脏脂肪合成通路上的由介导瘦素调节糖脂代谢平衡的关键效应基因之一。ACL位于细胞质中，能将胞质内的柠檬酸催化转变成为乙酰辅酶A，进而产生脂肪酸合成的前体分子-丙二酸单酰辅酶A，因此ACL是连接细胞内葡萄糖分解代谢与脂肪酸合成通路的重要代谢酶，是糖代谢与产生脂肪酸之间的桥梁(Elshourbagy et al.，Eur JBiochem 1992,204:491-499)。研究表明，ACL异常升高在糖脂代谢紊乱中扮演重要角色，ACL表达的改变能使细胞糖、脂代谢发生异常，导致机体发生肥胖症、Ⅱ型糖尿病、高脂血症、非酒精脂肪肝等代谢型疾病或癌症。
研究还表明代谢综合征(metabolic syndrome,MS)是以中心型肥胖、高血压、血脂异常、代谢异常等多种代谢异常集簇为内涵，以胰岛素抵抗为共同病理生理基础的一组临床症候群；近年来临床上MS的发病率呈增高趋势，严重侵害患者身心健康。
有关研究报道了II型糖尿病(non-insulin-dependent diabetes mellitus)和肥胖症(obesity)是严重威胁人类身体健康的内分泌紊乱性代谢类疾病，常伴随着外周组织细胞对胰岛素敏感性的降低(即胰岛素抵抗)，引起肌肉、肝脏及脂肪组织中的糖脂代谢平衡失调；研究表明基因沉默或抑制ACL能降低葡萄糖诱导的胰岛素分泌，游离脂肪酸的升高能抑制ACL活性从而导致胰腺细胞的应激和凋亡，脂肪酸和胆固醇合成离不开细胞质中的乙酰辅酶A参与；因此，ACL可视为治疗糖尿病和肥胖症疾病的重要药物靶点(Guay et al.,J Bio Chem 2007,282:35657-35665；Chu et al.,J Bio Chem 2010,285:32606–32615；Pearce et al.,Biochem J 1998,334:113–119)。
研究报道了高脂血症是由于全身脂肪代谢紊乱引起血浆中一种或几种脂质结构失衡的疾病，主要是血中总胆固醇(totalcholesterol,TC)或甘油三酯(triglyceride,TG)过高或高密度脂蛋白胆固醇(HDL-C)过低；ACL抑制剂能明显降低血浆中胆固醇和甘油三酯水平(Pearce et al.,Biochem J 1998,334,113-119)，因此，ACL可视为治疗高血脂症的重要药物靶点。
研究报道了非酒精性脂肪性肝病(Nonalcoholic Fatty Liver Disease，NAFLD)是指除外酒精和其他明确的损肝因素所致的肝细胞内脂肪过度沉积为主要特征的临床病理综合征，与胰岛素抵抗和遗传易感性密切相关的获得性代谢应激性肝损伤，研究显示，在非酒精性脂肪肝疾病中ACL是过度表达的，并导致甘油三酯和肝细胞脂肪变性(Wang etal.,Hepatology 2009,49:1166-1175)，因此，ACL也是治疗高血脂症的重要药物靶点。
寻找ACL的特异性抑制剂成为本领域技术人员的共识，通过抑制ACL的活性调节脂肪细胞中脂肪酸和胆固醇合成，对代谢紊乱疾病，如II型糖尿病和肥胖症、高血脂症或非酒精性脂肪肝的预防和治疗有着重要的应用前景。
研究认为，恶性肿瘤的发生与异常脂类代谢有关，异常脂类代谢途径参与肿瘤细胞的物质、能量合成、生长和耐药的信号转导等的过程；有研究发现ACL与数种肿瘤的恶性进程相关，且在多种肿瘤细胞中活性是上调的，ACY抑制可使体内和体外的绝大多数肿瘤细胞停止生长，因此，有关ACL的抑制剂可用于治疗或预防癌症或在癌症发展时用于减缓其进展。
麦吊云杉(Picea brachytyla)隶属松科(Pinaceae)云杉属(Picea)植物，是一种常绿乔木，成年后高达30余米，是我国特有的面临濒危灭绝的珍稀树种之一，于1992年10月被列为国家珍稀树种第一批保护树种(Fu et al.,China Plant Red Data Book,SciencePress:Beijing；New York,1992)，主要分布于我国秦岭、大巴山脉及四川北部，目前其化学成分和药理活性尚未有任何报道。
基于现有技术的现状和基础，本申请的发明人拟进行保护性地采集少许麦吊云杉植物样品研发其在制药中的新用途，具体的，本申请拟提供麦吊云杉提取物及其制备方法和在制药中的用途，尤其是作为ACL抑制剂在制备治疗ATP-柠檬酸裂解酶介导疾病药物中的用途。
发明内容
本发明的目的在于基于现有技术的现状和基础，提供麦吊云杉提取物及其制备方法和在制药中的用途，尤其是作为ACL抑制剂在制备治疗ATP-柠檬酸裂解酶介导疾病药物中的用途。本发明提供了一种麦吊云杉提取物，包括有粗提取物和单成分，及其作为ACL抑制剂的药物用途，可进一步用于制备治疗高血脂、心血管疾病及其他ACL介导的疾病的药物或保健品中应用。
本发明的进一步目的是提供了所述麦吊云杉提取物包括粗提取物和萜类单体化合物的制备方法。
本发明所述提取物由麦吊云杉枝叶经由本领域所涉常规的提取分离方法制备而得，其中粗提取物步骤可用下述方法制得：取干燥的麦吊云杉枝叶粉碎机粉粹；用8倍量的90％甲醇浸渍提取3次，每次24小时，得麦吊云杉提取液；麦吊云杉提取液用布氏漏斗抽滤，合并滤液，减压浓缩成浸膏，得麦吊云杉醇提物；其中萜类单体化合物采用以下方法制得：麦吊云杉粗提取物加适量水混悬后，先后用石油醚、乙酸乙酯和正丁醇萃取，分别得石油醚部分、乙酸乙酯部分和正丁醇部分，浓缩后真空干燥，分别得醇提物的石油醚部位、乙酸乙酯部位和正丁醇部位；取上述乙酸乙酯部位，经反复硅胶柱色谱、Sephadex LH-20凝胶色谱及反相半制备高效液相色谱(semi-RP-HPLC)分离制备，分别得到单体化合物brachytylinF(I),brachytylin G(II),23-hydroxyisomangiferolic acid A(III),abiesadine J(IV)，abietic acid(V)和isopimaric acid(VI)。
本发明所述萜类单体化合物结构如下所示：
Figure BDA0001968300450000031
本发明采用体外抑制ACL活性模型考察了麦吊云杉枝叶醇提物及醇提物石油醚部位、乙酸乙酯部分、正丁醇部分对ACL的抑制作用；并针对抑制活性最优的乙酸乙酯部分进行分离纯化，得到具有新化学结构和已知化学结构的萜类化合物，经体外多次生物活性测试研究表明，制得的植物粗提取物、有效部位及萜类化合物均具有抑制ACL的活性。
可进一步制备预防或治疗由ACL介导的糖脂代谢紊乱相关疾病，如高脂血症、动脉粥样硬化、非酒精性脂肪肝、Ⅱ型糖尿病、肥胖或癌症的药物或保健品。
本发明的优点是：
从麦吊云杉枝叶提取的麦吊云杉粗提取物以及从中分离得到的两个新三萜化合物(brachytylin F和brachytylin G)以及四个已知结构萜类化合物23-hydroxyisomangiferolic acid A,abiesadine J,abietic acid及isopimaric acid均可作为ACL抑制剂，对开发成为具有预防或治疗由ACL介导的糖脂代谢紊乱相关疾病，如高脂血症、动脉粥样硬化、非酒精性脂肪肝、Ⅱ型糖尿病、肥胖或癌症的药物或保健品，具有广阔的应用前景。
附图说明
图1为化合物23-hydroxyisomangiferolic acid A的1H NMR谱图。
图2为化合物abiesadine J的1H NMR谱图。
图3为化合物abietic acid的1H NMR谱图。
图4为化合物isopimaric acid的1H NMR谱图。
具体实施方式
下面结合具体实施例对本发明作进一步阐述，但这些实施例绝非对本发明有任何限制。本领域技术人员在本说明书的启示下对本发明实施方式中所作的任何变动都将落在权利要求书的范围内。
下述制备例中，比旋光测试通过JASCO P-1020旋光仪完成；紫外和红外光谱数据分别通过Shimadzu UV-2550紫外光谱仪和Nicolet AVATAR 360型红外光谱仪获得；NMR用Bruker Avance II 400型仪测定；ESI-MS由Agilent 1100Series LC/MSD G1946D型仪测定,HR-ESI-MS由AB Sciex TripleTOF 5600型仪测定；所使用的硅胶和薄层板为烟台康比诺公司生产，Sephadex LH-20凝胶为瑞士GE Healthcare Bio-Sciences公司生产；半制备HPLC为Waters e2695，配备2998PDA和2424蒸发光散射双检测器以及SunFire ODS(5μm,250×10mm)半制备柱；所有试剂均为上海国药集团化学试剂有限公司生产。
实施例1、制备麦吊云杉枝叶粗提取物
(1)取干燥的麦吊云杉枝叶800g，粉碎机粉碎；
(2)用8倍量的90％甲醇浸渍提取3次，每次24小时，得麦吊云杉醇提取液；
(3)麦吊云杉醇提取液用布氏漏斗抽滤，合并滤液，减压浓缩成浸膏，得麦吊云杉粗提取物120g。
实施例2、制备麦吊云杉萜类单体化合物
取麦吊云杉粗提取物110g加适量水混悬后，先后用石油醚、乙酸乙酯和正丁醇萃取，得石油醚部分、乙酸乙酯部分和正丁醇部分，浓缩后真空干燥，得醇提物的石油醚部分20g、乙酸乙酯部分32g和正丁醇部分24g。继而取麦吊云杉乙酸乙酯部分30g，以等倍硅胶(100～200目)拌样，进行柱层析，以石油醚：乙酸乙酯15:1-0:1及乙酸乙酯：甲醇15:1-0:1梯度洗脱，得到10个组分(Fr.1-Fr.10)；亚组分Fr.5继续过MCI柱色谱，采用水：甲醇(1:1-0:1)洗脱；对1:4洗脱组分采用Sephadex LH-20进一步纯化，重结晶得到isopimaric acid(10mg)(氢谱谱图如图4所示)，剩余部分经过半制备型HPLC进一步分离，得到化合物abietic acid(2.9mg)(氢谱谱图如图3所示)；
亚组分Fr.6继续过MCI柱色谱，采用水：甲醇(1:1-0:1)洗脱；对1:4洗脱组分采用Sephadex LH-20进一步纯化，经过半制备型HPLC进一步分离，分别得到化合物23-hydroxyisomangiferolic acid A(1.5mg)(氢谱谱图如图1所示),abiesadine J(3.1mg)(氢谱谱图如图2所示)，以及brachytylin F和brachytylin G的混合物(1.4mg)；
化合物23-hydroxyisomangiferolic acid A,abiesadine J,abietic acid,isopimaric acid依据与文献数据(MS、1H NMR、比旋光)对比，最终确证其结构；
化合物brachytylin F和brachytylin G为一对新的环阿屯烷三萜类化合物的C-24位差向异构体混合物(比例1:1)，光谱及理化数据如下：白色无定形粉末；[α]20 D+12.8(c0.14,MeOH)；UV(MeOH)λmax(logε)204(2.2)nm；IR(KBr)νmax 3400,3030,2950,2870,1680,1630,1450cm-1；1H-NMR(400MHz，CD3OD)：δ1.49(1H,m,H-1a),1.25(1H,m,H-1b),1.76(1H,m,H-2a),1.57(1H,m,H-2b),3.30(1H,dd,J＝10.8,4.8Hz,H-3),1.31(1H,m,H-5),1.60(1H,m,H-6a),0.80(1H,m,H-6b),1.34(1H,m,H-7a),1.10(1H,m,H-7b),1.50(1H,m,H-8),2.00(1H,m,H-11a),1.11(1H,m,H-11b),1.82(1H,m,H-12a),1.62(1H,m,H-12b),1.30(2H,m,H-15),1.90(1H,m,H-16a),1.31(1H,m,H-16b),1.60(1H,m,H-17),0.97(3H,s,H-18),0.55,0.34(each 1H,d,J＝4.0Hz,H-19a and H-19b),1.57(1H,m,H-20),0.88(3H,d,J＝6.0Hz,H-21),1.44(2H,m,H-22),1.75(2H,m,H-23),4.38(1H,m,H-24),6.37(1H,br s,H-27a),5.91/5.89(1H,br s,H-27b),0.81(3H,s,H-28),0.97(3H,s,H-29),0.89(3H,s,H-30)；13C-NMR(150MHz,CD3OD):δ32.0(C-1),30.4(C-2),78.9(C-3),40.5(C-4),47.1(C-5),21.1(C-6),26.0(C-7),48.0(C-8),20.0(C-9),26.1(C-10),26.4(C-11),32.9(C-12),45.3(C-13),48.8(C-14),35.5(C-15),28.1(C-16),52.2/52.1(C-17),18.0(C-18),29.9(C-19),32.2/32.1(C-20),18.3/18.2(C-21),35.9/35.7(C-22),33.0/32.9(C-23),72.8/72.1(C-24),141.5/141.2(C-25),168.2(C-26),127.2/126.9(C-27),14.0(C-28),25.4(C-29),19.3(C-30)；HRESIMS[M-H]-m/z 471.3483(calcd for C30H47O4,471.3480).。
实施例3、麦吊云杉粗提取物及萜类单体化合物对ATP柠檬酸裂解酶抑制活性实验
材料：ACL---人ACLY/acly/ATP citreate lyase蛋白(His标签)(SinoBiological公司购买)；激酶检测试剂盒ADP-GLO(Promega公司)；
操作步骤：将化合物、酶与底物37度共孵育半小时，加入ADP-Glo&#8482；试剂孵育半小时，以终止反应，并消耗完剩余的ATP；再加入激酶检测试剂(使ADP转化成ATP的同时，还使用偶联的萤光素酶/萤光素反应来检测新合成的ATP)孵育半小时用Envision读值)；初筛选择所述提取物浓度为20μg/mL时对ACL酶活性的百分抑制率进行考察，试验结果进一步测定IC50值：样品临用前加入10％DMSO和2.5mL测活体系稀释配成1％DMSO的反应液，酶反应的温度为37℃，时间为30min。酶反应结束后，加入25mL ADP-Glo试剂室温培育30min，随后加入激酶检测试剂室温培育30min，在EnVision上测量，其动力学曲线一级反应的斜率作为酶的活性指标；以相对活性对化合物浓度作图，经公式v/v0＝100/(1+b*[I]/IC50)拟合得到IC50值，实验重复三次，结果取三次的平均值；测试数据所表1所示：测试结果表明，麦吊云杉粗提取物及萜类单体化合物均对ACL均表现出较强抑制活性，表明所述的粗提取物及其萜类单体化合物可用于制备预防或治疗由ACL介导的糖脂代谢紊乱相关疾病，如高脂血症、动脉粥样硬化、非酒精性脂肪肝、Ⅱ型糖尿病、肥胖或癌症的药物或保健品，或作为该类药物的先导化合物。
表1.麦吊云杉粗提取物及其萜类单体化合物抑制ACL活性数据
Figure BDA0001968300450000061
a BMS 303141：Positive control。

Claims (6)

1.麦吊云杉提取物，其特征在于，所述的麦吊云杉提取物包括粗提取物和单一成分；其中单一成分为式Ⅰ～式V所示的萜类单体化合物brachytylin F(I),brachytylin G(II),23-hydroxyisomangiferolic acid A(III),abiesadine J(IV)，abietic acid(V)和isopimaric acid(VI)；
Figure FDA0001968300440000011
2.如权利要求1所述的麦吊云杉提取物，其特征在于，所述的粗提取物采用下述方法制得：取干燥的麦吊云杉枝叶粉碎机粉粹；用8倍量的90％甲醇浸渍提取3次，每次24小时，得麦吊云杉提取液；麦吊云杉提取液用布氏漏斗抽滤，合并滤液，减压浓缩成浸膏，得麦吊云杉醇提物。
3.如权利要求1所述的麦吊云杉提取物，其特征在于，所述的单一成分采用下述方法制得：麦吊云杉粗提取物加适量水混悬后，先后用石油醚、乙酸乙酯和正丁醇萃取，分别得石油醚部分、乙酸乙酯部分和正丁醇部分，浓缩后真空干燥，分别得醇提物的石油醚部位、乙酸乙酯部位和正丁醇部位；取上述乙酸乙酯部位，经反复硅胶柱色谱、Sephadex LH-20凝胶色谱及反相半制备高效液相色谱(semi-RP-HPLC)分离制备，分别得到单体化合物brachytylin F(I),brachytylin G(II),23-hydroxyisomangiferolic acid A(III),abiesadine J(IV)，abietic acid(V)和isopimaric acid(VI)。
4.权利要求1所述的麦吊云杉提取物在制备ATP-柠檬酸裂解酶ACL抑制剂药物中的用途。
5.如权利要求4所述的用途，其特征在于，所述的药物为预防或治疗ACL介导的糖脂代谢紊乱相关疾病的药物或保健品；所述糖脂代谢相关代谢疾病包括高脂血症、动脉粥样硬化、非酒精性脂肪肝、II型糖尿病及肥胖症。
6.如权利要求1所述的麦吊云杉提取物在制备预防或治疗肿瘤的药物或保健品中用途。
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113563409A (zh) * 2021-06-25 2021-10-29 台州学院 天然三萜-环烯醚萜苷二聚体杂合物及其制备和在制备acl抑制剂中的用途

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2016029322A1 (en) * 2014-08-29 2016-03-03 Borea (9302-0469 Québec Inc.) High grade spruce essential oil, method for manufacturing high grade spruce essential oil and apparatus for manufacturing high grade spruce essential oil

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2016029322A1 (en) * 2014-08-29 2016-03-03 Borea (9302-0469 Québec Inc.) High grade spruce essential oil, method for manufacturing high grade spruce essential oil and apparatus for manufacturing high grade spruce essential oil

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
刘家仪等: "ATP-柠檬酸裂解酶研究进展", 《南方农业学报》 *
江维等: "秦岭地区特有松科植物麦吊云杉的化学成分研究", 《中国化学会第十二届全国天然有机化学学术会议论文摘要集》 *
白艳等: "贡嘎山和昆仑山表土中甲氧基脂肪酸的生态指示意义", 《科学通报》 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113563409A (zh) * 2021-06-25 2021-10-29 台州学院 天然三萜-环烯醚萜苷二聚体杂合物及其制备和在制备acl抑制剂中的用途

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