CN109111387B - 一种氨基酸类化合物、应用及制备方法 - Google Patents
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- C07D207/22—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having one double bond between ring members or between a ring member and a non-ring member with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
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Abstract
本发明提供一种氨基酸类化合物的制备方法,包括以下步骤:用乙醇提取黄精粉末,得到的提取液用二氯甲烷萃取,萃取层浓缩得到浸膏;浸膏用二氯甲烷‑甲醇进行第一次洗脱,收集得到第一次洗脱后的活性组分;第一次洗脱后的活性组分经柱色谱第二次洗脱,收集第二次洗脱后的活性组分经柱色谱第三次洗脱,收集第三次洗脱后的活性组分经柱色谱第四次洗脱,洗脱液纯化得到氨基酸类化合物。本发明首次从黄精中提取出氨基酸类化合物,并且研究得出其在抗肝癌及抗乳腺癌方面有显著的效果,具有重要的临床应用价值。本发明方法所使用的有机溶剂毒性较小,易于回收;且本方法具有工序少、设备简单、操作方便、可行性好、生产成本低廉等特点。
Description
技术领域
本发明涉及药物提取技术领域,具体涉及到一种氨基酸类化合物、应用及制备方法。
背景技术
多花黄精,来源于百合科黄精属多花黄精植物的干燥根茎,又名姜形黄精。主产于我国南方地区,湖南省栽培多花黄精历史悠久,是其道地产地之一。《本草纲目》言其“得坤土之精,为补养中宫之胜品”,具有补肾益精,滋阴润燥的功效,长期用于治疗肾虚亏损,脾胃虚弱,肺虚燥咳,体倦乏力之症,同时也是数十种复方滋补药剂的重要组分,黄精根状茎形状有如山芋,山区老百姓常把它当作蔬菜食用药用植物,具有补脾,润肺生津的作用。此外,由于黄精还含有多种功效成分,如呋甾烯醇型皂甙、螺甾烯醇型皂甙和多糖等,通常还用于抗氧化、抗疲劳以及抗菌方面。
目前已经从黄精中提取了多种活性物质,如专利申请号CN104069348A公开了一种黄精提取物及其制备方法与应用,该发明将黄精通过乙醇热提后,再用石油醚、乙酸乙酯、正丁醇萃取,得到的化合物中主要含5-羟甲基糠醛、甾体皂苷和酚酸类(木脂素等)成分,主要用于抗糖尿病及其并发症上;专利申请号CN201510395570.X公开了一种黄精中多种活性成分的提取分离方法,通过闪式提取技术提取分离黄精多糖、黄精皂苷和黄精黄酮等活性成分。目前从黄精中提取分离出焦谷氨酸甲酯至今尚未见报道。
焦谷氨酸甲酯属于已经公开的化合物,有人从红树药用植物角果木、味噌、短瓣花等植物中分离提纯出焦谷氨酸甲酯,但是对其在抗肝癌和乳腺癌中的应用尚没有研究。
发明内容
本发明的目的是克服现有技术的不足,提供一种氨基酸类化合物、应用及制备方法。
本发明提供了一种氨基酸类化合物,所述氨基酸类化合物的结构式如下:
本发明提供了一种氨基酸类化合物的应用,所述氨基酸类化合物用于制备抗肝癌和/或抗乳腺癌的产品。
本发明还提供了一种氨基酸类化合物的制备方法,包括以下步骤:
(1)用乙醇提取黄精粉末,得到的提取液用二氯甲烷萃取,二氯甲烷层浓缩得到浸膏;
(2)浸膏经柱色谱用二氯甲烷-甲醇进行第一次洗脱,收集得到第一次洗脱后的活性组分;
(3)第一次洗脱后的活性组分经柱色谱第二次洗脱,收集第二次洗脱后的活性组分经柱色谱第三次洗脱,收集第三次洗脱后的活性组分经柱色谱第四次洗脱,收集第四次洗脱后的洗脱液纯化得到氨基酸类化合物。
优选的,所述步骤(1)中的黄精粉末的制备方法为:将多花黄精根部经过干燥和粉碎处理,过筛后得到黄精粉末。
优选的,所述步骤(1)中乙醇的体积浓度为95%。
优选的,所述步骤(1)中黄精粉末与乙醇的质量比为1:5-8。
优选的,所述步骤(2)中的柱色谱为硅胶柱色谱。
优选的,所述步骤(2)中二氯甲烷-乙醇的体积比为1:0-6:4。
优选的,所述步骤(3)中第二次洗脱的洗脱剂为石油醚-氯仿。
优选的,所述步骤(3)中第三次洗脱的洗脱剂和第四次洗脱的洗脱剂均为氯仿-甲醇。
优选的,所述步骤(3)中第四次洗脱所用的柱色谱为葡聚糖凝胶柱。
本发明所述活性氨基酸类化合物可用于制备抗肝癌及抗乳腺癌的药品和保健品。
本发明所述步骤(1)中所述用乙醇提取黄精粉末分为多次提取,以最大限度提取出原料中的活性成分,提取次数不少于2次。
本发明的黄精粉末用二氯甲烷萃取后得到的浸膏上样于硅胶柱色谱,用体积比为1:0-6:4的二氯甲烷-甲醇进行第一次梯度洗脱,收集洗脱比例为70%(即二氯甲烷与甲醇的体积比为7:3)的洗脱液,即为第一次洗脱后的活性组分;取第一次洗脱后的活性组分上样于硅胶柱色谱,用体积比为95:5-0:1的石油醚-氯仿进行第二次梯度洗脱,收集洗脱比例为60%(即石油醚与氯仿的体积比为3:2)的洗脱液,即为第二次洗脱后的活性组分;取第二次洗脱后的活性组分上样于硅胶柱色谱,用体积比为95:5-0:1的氯仿-甲醇进行第三次梯度洗脱,收集洗脱比例为60%(即氯仿与甲醇的体积比为3:2)的洗脱液,即为第三次洗脱后的活性组分;取第三次洗脱后的活性组分上样于葡聚糖凝胶柱,用体积比为1:1的氯仿-甲醇洗脱,得到的洗脱液进一步纯化,得到氨基酸类化合物(焦谷氨酸甲酯)。
本发明所述梯度洗脱时洗脱剂的极性由小到大。
本发明所述氨基酸类化合物为焦谷氨酸甲酯。
目前,研究人员从黄精中提取了多种活性成分,如黄精多糖、皂苷和黄酮等,主要运用的方法是醇提或者是水提,再将提取液用有机溶剂或水进行多次萃取,有机层再经过洗脱,纯化得到活性成分。但是常用的萃取剂是石油醚、乙酸乙酯和正丁醇,如专利申请号CN104069348A中依次用上述3中溶剂萃取,专利申请号CN201810125885.6中用乙酸乙酯萃取,专利申请号CN201510395570.X中用正丁醇萃取;尚未出现用二氯甲烷做萃取剂来提取黄精中的活性物质,本领域的技术人员知道萃取剂不同,萃取后得到的物质也会不同。二氯甲烷与石油醚、乙酸乙酯和正丁醇属于不同类的物质,结构和性质都有较大的区别,因此,在现有技术的基础上,本领域技术人员根据使用石油醚、乙酸乙酯和正丁醇做萃取剂,不容易想到用二氯甲烷代替上述有机溶剂。
本发明萃取后得到的浸膏经过多次洗脱,纯化得到氨基酸类化合物,实现了从黄精中提取出焦谷氨酸甲酯。本发明所用的洗脱剂与现有技术中所用的洗脱剂有较大的区别,现有技术中使用较多的洗脱剂是水、乙醇或石油醚-乙酸乙酯,如专利申请号CN201711483544.8中用石油醚-乙酸乙酯为洗脱剂洗脱进行梯度洗脱,提取黄精皂苷。并且在其他中药物质的提取中,大都是一次洗脱,而且就算是多次洗脱其洗脱剂也相差不大,并未出现以石油醚-氯仿混合进行洗脱,而是以单一的石油醚或者氯仿洗脱,本领域技术人员知道以两种溶剂混合洗脱相比于单一的有机溶剂洗脱,其效果是不可预测的,因此现有技术中有以单一的石油醚或者氯仿做洗脱剂,并不能想到将两者混合做洗脱剂;而本发明三次梯度洗脱的洗脱剂都不同,第一次洗脱是二氯甲烷-甲醇,第二次洗脱是石油醚-氯仿,第三次洗脱是氯仿-甲醇,所用的洗脱剂与现有用于黄精活性成分提取的洗脱剂有较大的差别,而洗脱剂的种类不同,极性不同,最终黄精中提取得到的活性成分也不一样,因此,在现有技术的基础上,并不存在任何的启示将现有技术中的洗脱剂替换为本申请中的洗脱剂,也就难以得到本申请的技术方案。
本申请用乙醇提取黄精粉末,得到的提取液用二氯甲烷萃取,浓缩得到浸膏;浸膏用二氯甲烷-甲醇第一次洗脱,收集第一次洗脱后的活性组分用石油醚-氯仿第二次洗脱,收集第二次洗脱后的活性组分用氯仿-甲醇第三次洗脱,收集第三次洗脱后的活性组分用氯仿-甲醇第四次洗脱,得到的洗脱液纯化得到焦谷氨酸甲酯,这种化合物是首次在黄精中被提取出来,制备方法简单,操作方便。
虽然目前已有从黄精中提取出具有抗癌作用的化合物,但是其主要应用于结肠癌、H22实体瘤和S180腹水瘤,与本发明化合物的应用范围并不相同。
虽然焦谷氨酸甲酯属于已经公开的化合物,有人从红树药用植物角果木、味噌、短瓣花等植物中分离提纯出焦谷氨酸甲酯,但是对其在抗肝癌和乳腺癌中的应用尚没有研究。运用本发明的方法从黄精中提取出氨基酸类物质,即焦谷氨酸甲酯,本申请的发明人通过研究发现焦谷氨酸甲酯能有效抗肝癌及抗乳腺癌,能用于制备药品和保健品。
本发明的有益效果是:
1、本发明用乙醇提取黄精粉末,得到的提取液用二氯甲烷萃取,浓缩得到浸膏;浸膏用二氯甲烷-甲醇第一次洗脱,收集第一次洗脱后的活性组分用石油醚-氯仿第二次洗脱,收集第二次洗脱后的活性组分用氯仿-甲醇第三次洗脱,收集第三次洗脱后的活性组分用氯仿-甲醇第四次洗脱,得到的洗脱液纯化得到焦谷氨酸甲酯,这种化合物是首次在黄精中被提取出来,充分利用了黄精资源,提高了原材料的使用价值,对推进黄精皂苷的研究、应用有重要意义。
2、本发明首次从黄精中提取出焦谷氨酸甲酯,并且研究得出其在抗肝癌及抗乳腺癌方面有显著的效果,具有重要的临床应用价值。
3、本发明方法所使用的有机溶剂毒性较小,易于回收;且本方法具有工序少、设备简单、操作方便、可行性好、生产成本低廉等特点。
附图说明
图1为本发明的氨基酸类化合物的13C-NMR谱。
图2为本发明的氨基酸类化合物的1H-NMR谱。
具体实施方式
为使本发明的目的、技术方案和优点更加清楚明白,以下结合具体实施例,对本发明进一步详细说明。
实施例1
(1)取500g多花黄精根部原料,干燥后经粉碎处理,过筛得到黄精粉末,用2500ml体积浓度为95%的乙醇提取,提取使用仪器为DTQ-200型多功能提取罐,重复提取2次后,得到提取液;将提取液用2500ml的二氯甲烷进行萃取,二氯甲烷层浓缩得到浸膏;
(2)将浸膏上样于硅胶柱,用1600ml体积比为1:0-6:4的二氯甲烷-甲醇进行第一次梯度洗脱,收集洗脱比例为70%的洗脱液,即为第一次洗脱后的活性组分;
(3)取第一次洗脱后的活性组分上样于硅胶柱色谱,用3000ml体积比为95:5-0:1的石油醚-氯仿进行第二次梯度洗脱,收集洗脱比例为60%的洗脱液,即为第二次洗脱后的活性组分;取第二次洗脱后的活性组分上样于硅胶柱色谱,用体积比为95:5-0:1的氯仿-甲醇进行第三次梯度洗脱,收集洗脱比例为60%的洗脱液,即为第三次洗脱后的活性组分;取第三次洗脱后的活性组分上样于葡聚糖凝胶柱,用体积比为1:1的氯仿-甲醇洗脱,得到的洗脱液进一步纯化,得到氨基酸类化合物,其纯度大于90%。
经鉴定,该氨基酸类化合物的核磁共振谱图如表1所示,核磁共振碳谱和核磁共振氢谱如图1和2所示。
表1氨基酸类化合物的核磁谱图
通过分析波谱数据并与陈权威等在军事医学发表的《味噌含氮类化学成分研究》和程永现等在云南植物研究学报发表的《短瓣花中的含氮化合物》对照,确定该氨基酸类化合物为焦谷氨酸甲酯。
实施例2
(1)取500g多花黄精根部原料,干燥后经粉碎处理,过筛得到黄精粉末,用4000ml体积浓度为95%的乙醇提取,提取使用仪器为DTQ-200型多功能提取罐,重复提取3次后,得到提取液;将提取液用2000ml的二氯甲烷进行萃取,二氯甲烷层浓缩得到浸膏;
(2)将浸膏上样于硅胶柱,用1600ml体积比为1:0-6:4的二氯甲烷-甲醇进行第一次梯度洗脱,收集洗脱比例为70%的洗脱液,即为第一次洗脱后的活性组分;
(3)取第一次洗脱后的活性组分上样于硅胶柱色谱,用3000ml体积比为95:5-0:1的石油醚-氯仿进行第二次梯度洗脱,收集洗脱比例为60%的洗脱液,即为第二次洗脱后的活性组分;取第二次洗脱后的活性组分上样于硅胶柱色谱,用体积比为95:5-0:1的氯仿-甲醇进行第三次梯度洗脱,收集洗脱比例为60%的洗脱液,即为第三次洗脱后的活性组分;取第三次洗脱后的活性组分上样于葡聚糖凝胶柱,用体积比为1:1的氯仿-甲醇洗脱,得到的洗脱液进一步纯化,得到氨基酸类化合物,其纯度大于90%。
1、焦谷氨酸甲酯抗肿瘤活性研究
为检测焦谷氨酸甲酯对不同肿瘤细胞增殖的影响,现做以下抗肿瘤实验。
1.1构建细胞培养及相关细胞库平台
在胎牛血清中培养不同种类的癌细胞,其中不同癌细胞与编号的对应关系如表2所示;
表2不同癌细胞和与之对应编号的关系
1.2实验部分
1.2.1 MTT法分析焦谷氨酸甲酯对肿瘤细胞增殖的影响
抗肿瘤实验的原理为:活细胞的线粒体内膜上存在琥珀酸脱氢酶,该酶可将黄绿色的噻唑蓝(简称为MTT,为一种接受氢离子的染料)降解成蓝紫色的甲臜,活细胞越多,生成的蓝紫色的甲臜就越多,而死细胞因其线粒体内膜上的琥珀酸脱氢酶活性消失,无此功能;使用二甲基亚砜溶解蓝紫色的甲臜,并用酶标仪在490nm波长处测定吸光度值,可以定量反应出活细胞数量。
1.2.2焦谷氨酸甲酯抑制细胞增殖实验
(1)实验分组:将实验分为实验组(焦谷氨酸甲酯)和对照组,其中对照组为紫杉醇,不同组的药物均设置有4个浓度梯度的加药,以上每组均设有3个复孔;
(2)细胞培养:取对数生长期的HepG-2、Hela、MCF-7及BGC-823细胞,吸出原培养液,无菌PBS洗涤两次后加入适量的胰酶进行消化,并离心收集细胞,制成细胞悬液;对细胞进行计数,并稀释细胞浓度为6x103/mL,准备无菌的96孔细胞培养板并进行相应的标记,然后每孔加入100μL细胞悬液,置于37℃,CO2体积浓度为5%的培养箱内培养。待细胞贴壁生长至80%左右时,实验组每孔加入相应的药物,置于37℃,CO2体积浓度为5%的培养箱内培养24h。培养48h后,吸出原培养液,无菌PBS洗涤两次后每孔加入含0.5mg/mLMTT的培养基继续培养4h。4h后,小心吸出孔内培养基,每孔加入150μL二甲基亚砜,置摇床上匀速摇10min,使结晶充分溶解;再用酶标仪在490nm波长处测定各组吸光度值,计算细胞增殖抑制率。
1.3结果与分析
1.3.1 MTT实验检测结果
运用焦谷氨酸甲酯作用于肿瘤细胞24h后,酶标仪测定各个孔吸光度,并计算细胞增殖抑制率,结果见表3。
表3焦谷氨酸甲酯对肿瘤细胞活性的影响
其中,IC50(μM)为增殖抑制率为50%时焦谷氨酸甲酯的浓度,用于表示抗肿瘤活性;紫杉醇为阳性对照药;NC表示浓度大于25μM时,焦谷氨酸甲酯依旧无明显抗肿瘤活性。
如表3所示,焦谷氨酸甲酯以剂量依赖性方式抑制HepG-2和MCF-7肿瘤细胞的增殖,其IC50结果显著低于10μM,表明焦谷氨酸甲酯对HepG-2和MCF-7肿瘤细胞有较好的抑制作用。
而Hela和BGC-823肿瘤细胞对焦谷氨酸甲酯的浓度变化反应不明显,因此焦谷氨酸甲酯对Hela和BGC-823肿瘤细胞的抑制作用并没有其对HepG-2和MCF-7肿瘤细胞的抑制作用好。
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Non-Patent Citations (6)
Title |
---|
"Antibacterial and Antioxidant Activities of Quercetin Oxidation Products from Yellow Onion (Allium cepa) Skin";FREDDY A. RAMOS, et al.,;《J. Agric. Food Chem.》;20061231;第54卷(第10期);3551-3557 * |
"Anticancer Activity of Sida cordifolia L., – Insilico approach";Meenakshi Sundaram Muthuraman et al;《J. Pharm. Sci. & Res.》;20171231;第9卷(第8期);1363-1367 * |
"L-焦谷氨酸酯的合成及应用";彭小海等;《化学与生物工程》;20111031;第28卷(第10期);79-80 * |
"Odisolane, a Novel Oxolane Derivative, and Antiangiogenic Constituents from the Fruits of Mulberry (Morus alba L.)";Seoung Rak Lee et al.,;《J. Agric. Food Chem.》;20160426;3804-3809 * |
"Pyrrolidin-2-one derivatives from Iris pseudacorus (Iridaceae)";F. Senatore et al.,;《Biochemical Systematics and Ecology》;20031231;657-659 * |
"黄精乙酸乙酯部位的化学成分研究";陈辉等;《中药材》;20170630;第40卷(第6期);1345-1347 * |
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