CN101787061A - Application of Quzhazhigan in preparation of preparations for preventing and treating cardiac-cerebral ischemia diseases, and preparation method thereof - Google Patents

Application of Quzhazhigan in preparation of preparations for preventing and treating cardiac-cerebral ischemia diseases, and preparation method thereof Download PDF

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CN101787061A
CN101787061A CN 201010116358 CN201010116358A CN101787061A CN 101787061 A CN101787061 A CN 101787061A CN 201010116358 CN201010116358 CN 201010116358 CN 201010116358 A CN201010116358 A CN 201010116358A CN 101787061 A CN101787061 A CN 101787061A
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stilbene
sheaf
extract
glycosides
song
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CN101787061B (en )
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代家红
何金星
胡琳
胡群
胡茂
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昆明翔昊科技有限公司
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Abstract

The invention relates to application of Quzhazhigan serving as polyhydroxy stilbene glycoside compounds in preparation of preparations for preventing and treating ischemic cardio-cerebrovascular diseases, and a preparation method thereof. The Quzhazhigan is compounds (E)-1-(3,5-dihydroxy phenyl)-2(3-hydroxyl-4-O-beta-D-pyran glucose phenyl)ethane, and has a chemical structural formula. Test results show that the Quzhazhigan can significantly reduce injury caused by ischemia reperfusion after cardio-cerebral infarction and has wide application in the preparation of preparations for preventing and treating cardiac-cerebral ischemia diseases. The Quzhazhigan can be extracted from roots and/or rhizome of Rheum lhasaense and prepared in pure alcohol/water systems, thereby having the advantagesof simple convenient property, environment-friendly property and strong practicability.

Description

曲札疾苷在制备防治心脑缺血疾病制剂中的应用及其制备 Application and Qu sheaf disease glycosides in Preparation prevention of ischemic heart and brain diseases formulation

方法 method

技术领域 FIELD

[0001] 本发明涉及植物活性成分曲札芪苷在制备防治心脑缺血制剂中的用途及曲札芪苷、曲札芪苷提取物的一种制备方法。 [0001] The present invention relates to a method for preparing plant active ingredient sheaf stilbene glycosides song and song sheaf use in the manufacture of stilbene glycosides prevention of ischemic heart and brain formulation, Sapporo curved stilbene glucoside extract. 所述曲札芪苷为化合物(E)-l-(3,5-二羟苯基)-2- (3-羟基-4-0-D-吡喃葡萄糖苯基)乙烯,其制备方法是从拉萨大黄的根和/或根茎中提取,通过纯化步骤制备。 The curved sheaf stilbene glucoside compound (E) -l- (3,5- dihydroxyphenyl) -2- (3-hydroxy--4-0-D- glucopyranose phenyl) ethylene, which is prepared la is extracted from rhubarb roots and / or rhizomes, prepared by a purification step.

背景技术 Background technique

[0002] 曲札芪苷为化合物(E)-l_(3,5- 二羟苯基)-2-(3-羟基-4_0-|3 _D_吡喃葡萄糖苯基)乙烯,或称为3,5,3' ,4'-四羟基芪_3' -OP-葡萄糖苷,因其植物来源拉萨大黄根茎在藏药材中又名曲札(罗达尚主编。新修晶珠本草,四川科学技术出版社,2004),故称该化合物为曲札芪苷。 [0002] Qu sheaf stilbene glucoside compound (E) -l_ (3,5- dihydroxyphenyl) -2- (3-hydroxy--4_0- | 3 _D_ glucopyranosyl phenyl) ethylene, otherwise known as 3 , 5,3 ', 4'-tetrahydroxy-stilbene _3' -OP- glucoside, because plant sources in Lhasa Tibetan medicine rhubarb roots in music and Sapporo (Rhoda still editor of the newly built crystal beads Materia Medica, Sichuan Science and technology Press, 2004), so that the compound is a song Sapporo stilbene glycosides. 芪类物质是一类由二苯乙烯(图1)作为母体结构的衍生物,并由Gorham(Gorham J. Progress in Phytochemistry, Pergamon Press :0xford,1980, Vol. 6, pp. 203-252)于1980年正式命名为芪(Stilbenoids)。 Stilbene-based material is a class of stilbene (FIG. 1) as the derivative of a parent structure by Gorham (Gorham J. Progress in Phytochemistry, Pergamon Press:.. 0xford, 1980, Vol 6, pp 203-252) in 1980 officially named Qi (Stilbenoids). 芪类物质作为植物防御素多存在种子植物中,随着研究的深入,其化学、生物学多样性和不断新发现的生物活性受到了人们的 Stilbene as a plant substance there is more than defensin seed plants, with further research, its chemical, biological diversity and constant new discoveries of biological activity by the people

重视。 Seriously. 芪的母体结构式如下: [0003] Stilbene parent structural formula: [0003]

[0004] 90年代以后,以白藜戸醇(Resveratrol)为代表的天然芪类成分的药理活性研究受到广泛关注。 [0004] Since the 1990s, the pharmacological activity of quinoa to Kobe alcohol (Resveratrol), represented by stilbene natural ingredients attracted widespread attention. 国际上对如白藜戸醇、紫檀芪(Pterostilbene)等芪的药理作用研究主要集中于抗氧化、抗肿瘤以及植物雌激素活性等方面。 Alcohols such as C. quinoa Kobe, research stilbene pharmacological effects pterostilbene (Pterostilbene) focused on the anti-oxidation, anti-tumor activity of phytoestrogens and other aspects of the world. 国内在芪苷类成分如虎杖苷(Polydatin, piceid)、大黄降月旨素(Rhapontin, rhaponticin, 土大黄苷)、二苯乙烯苷的降血脂、降血糖以及心脑血管活性方面进行一系列研究。 Domestic as polydatin (Polydatin, piceid),, stilbene glycoside fat, lowering blood sugar and cardiovascular activity of a series of studies in the stilbene glycosides rhubarb month drop purpose hormone (Rhapontin, rhaponticin, rhaponticin) . 芪及芪苷类物质在多方面凸显出的潜在应用价值受到越来越多的关注。 Qi Qi glycosides substances and attracted more and more attention in the potential value of many highlights. 如中国专利申请99806755. 5及02806583. 2均公开了含有芪类化合物的复合抗癌剂,中国专利申请99122378. 0公开了波叶组大黄所含芪苷化合物的降糖活性;中国专利申请01118193.5公开了波叶组大黄所含芪苷类化合物的降血脂活性;中国专利01134283. 8公开了提取自何首乌的芪苷类化合物二苯乙烯甙的抗心脑缺血作用。 The Chinese Patent Application No. 99806755.5 and 02806583.2 disclose a composite containing a stilbene compound of the anticancer agent, Chinese Patent Application No. 99122378.0 discloses hypoglycemic activity of the wave leaf group stilbene glucoside compounds contained Rheum; Chinese Patent Application No. 01118193.5 discloses hypolipidemic activity contained in the wave leaf group Rheum stilbene glycosides; and Chinese Patent No. 01134283.8 discloses an anti-ischemic effect heart extracted from Polygonum stilbene glycosides stilbene glycosides brain.

[0005] 曲札芪苷于1984年(Y. Kashiwada, et al. Isolation andCharacterization of Stilbenes. Chem. Pharm. Bull. 1984,32(9) :3501-3517)首次从芋大黄(商品名:lmo-Diao) 中得到并进行了结构确认,得率约为0.4%,此后,有报道从河套大黄中分离得到少量该化合物(李军林等。河套大黄非蒽醌类成分研究,中草药,98,29:721),华北大黄中也含有该化合物,得率约为1.8% (王爱芹等。华北大黄中芪类成分研究,中草药,2001,32:878)。 [0005] Qu sheaf stilbene glycosides 1984 (.... Y. Kashiwada, et al Isolation andCharacterization of Stilbenes Chem Pharm Bull 1984,32 (9):. 3501-3517) for the first time from Conus rhubarb (trade name: lmo- diao) obtained to confirm structure and a yield of about 0.4%, and thereafter, it is reported that the compound was isolated as a small amount (Li Junlin from other constituents of the Loop Loop rhubarb rhubarb class of non-anthraquinone, herbs, 98,29: 721 ), also contains the North rhubarb compounds, yield of about 1.8% (Wang Aiqin North et Rheum stilbene constituents in herbal medicine, 2001,32: 878). 目前尚无曲札芪苷相关药理活性方面的研究或报道。 Current research related to aspects of the pharmacological activity is no song Sapporo stilbene glycosides or reported.

[0006] 曲札芪苷的糖苷取代位于非间位羟基取代的苯环上,与大黄降脂素、去氧土大黄苷、虎杖苷及何首乌中得到的系列二苯乙烯苷等芪苷类化合物的糖苷取代位置有结构上的差异。 [0006] Qu sheaf substituted stilbene glycosides glycosides position located between the non-hydroxy-substituted benzene ring, compound Rhapontinum, deoxy rhaponticin, Polygonum polydatin and obtained a series of stilbene glycosides stilbene glycosides there are structural differences glycoside substitution position. 与大黄降脂素、去氧土大黄苷、虎杖苷相比,曲札芪苷具有较多的酚羟基,本发明通过清除DPra自由基试验发现曲札芪苷的自由基清除能力高于上述化合物。 And Rhapontinum, deoxy rhaponticin than polydatin, Sapporo stilbene glycosides song more phenolic hydroxyl group, the present invention is by testing found song clearing DPra radical sheaf stilbene glycosides radical scavenging ability than the above compound . 自由基是缺血再灌注时形成的,是缺血组织损伤的重要根源,抗氧化剂和自由基清除剂可减轻缺血组织的损伤,曲札芪苷强的自由基清除活性使其在保护缺血心脑组织损伤中更具优势。 Free radicals are formed when ischemia and reperfusion of ischemic tissue injury is a major source of antioxidants and free radical scavengers reduce ischemic tissue damage, strong song Sapporo stilbene glycoside radical scavenging activity in protecting lack of it tissue damage in heart and brain blood advantage. 本发明人首次对来源于拉萨大黄的曲札芪苷提取物及其纯品进行了系列防治缺血性心脑血管疾病方面的药理和药效研究。 The present inventors have for the first time pharmacology and efficacy studies series prevention of ischemic heart disease and cerebrovascular aspects of Lhasa rhubarb song from Sapporo stilbene glycoside extracts and pure conducted. 研究采用两种与人类疾病相近的动物模型,测定多种药效学指标, 综合评价两种样品对急性心脑缺血的影响。 Research in two animal models of human disease similar measure multiple pharmacodynamic index, comprehensive evaluation of the impact of the two samples of acute myocardial and cerebral ischemia. 试验结果表明,曲札芪苷提取物和纯品能明显降低冠状动脉结扎致心肌缺血大鼠血清CK-MB及LDH的释放,縮小心肌梗死范围,抑制血小板聚集,提示其对心肌缺血有明显保护作用。 The results show that, Qu Sapporo stilbene glucoside extract and pure significantly reduced coronary artery ligation-induced myocardial ischemia release serum CK-MB and LDH, reduced myocardial infarct size, inhibiting platelet aggregation, suggesting that there is on Myocardial Ischemia protective effect. 曲札芪苷提取物和纯品可以明显减少MCAO大鼠脑梗塞范围,使其行为障碍程度得到明显改善,对缺血性心脑血管疾病的具有显著的保护作用。 Qu sheaf stilbene glucoside extracts and pure MCAO can significantly reduce infarct size in rats, it significantly improved degree of behavioral disorders, it has a significant protective effect on ischemic cardiovascular and cerebrovascular diseases. 因此,曲札芪苷提取物及其纯品在防治心脑缺血方面具有潜在的临床应用价值。 Therefore, the song Sapporo stilbene glycoside extracts and pure potential clinical value in the prevention and treatment of cardio-cerebral ischemia. [0007] 曲札芪苷不同的结构还带来化合物单体水溶性的优势。 [0007] stilbene glycosides sheaf different song structure also brings the advantage of a water-soluble monomer compounds. 芪类物质结构中苯环上羟基的增加和葡糖基的取代使化合物的水溶性增加,如白藜芦醇在水中溶解度是O. 03g/ L(标准状况:25°C, 100kPa),虎杖苷因带有糖基使其在水中的溶解度增加到约0. 5g/L,比白藜芦醇多一个羟基的白杉皮醇(Piceata皿ol)水溶解度为0. 5g/L。 Hydroxy substituent on the phenyl ring increases the material structure and stilbene glucosyl group increases the water-soluble compounds, such as resveratrol solubility in water is O. 03g / L (standard conditions: 25 ° C, 100kPa), Polygonum cuspidatum because it glycosyl glycoside having increased to about 0. 5g / L, more than one more hydroxyl Shirasugi resveratrol cortisol (Piceata dish oL) a water solubility of 0. 5g / L of solubility in water. 而结构中苯环上甲氧基的取代使化合物水溶性大大降低。 The substituent on the benzene ring structure methoxy-soluble compound is greatly reduced. 如大黄降脂素和去氧土大黄苷(甲基虎杖苷)的水溶性小于0.04g/L。 The Rhapontinum deoxy rhaponticin and (meth polydatin) water solubility of less than 0.04g / L. 曲札芪苷结构中带有糖基和较多的酚羟基,其溶解度约为1. lg/L,为其制作成注射、口服液等制剂应用带来便利。 Qu sheaf stilbene glycosides having sugar structure, and more phenolic hydroxyl groups, a solubility of about 1. lg / L, for convenience made into preparations for injection application, oral and the like.

[0008] 芪类物质如白藜戸醇可从虎杖(Polygonum cuspidatum Sieb. et Zucc)中提取纯化,也可进行合成,而芪苷类物质多从植物资源中提取。 [0008] stilbene extractable material, such as Chenopodium album Kobe alcohol from polygonum (Polygonum cuspidatum Sieb. Et Zucc) purified, can be synthesized, and multi-stilbene glycosides extracted from plant material resources. 研究表明,寥科植物(Polygonaceae),特别是大黄属(Rheum)植物中常含有较高的芪苷类成分,如河套大黄(Rheum hotaoense CYCheng et CT Kao)的根茎中土大黄苷的含量可达3% (逯海龙等,HPLC法测定栽培河套大黄中土大黄苷的含量,药物分析杂志,2007,27 :ll),大黄降脂素在部分地区栽培的唐古特大黄(Rhe咖tangutic咖Maxim, ex Balf.)根茎中的含量可达到10% (张德等,天然产物研究与开发,2005,17 :217)。 Studies have shown that very few plants (Polygonaceae), in particular rhubarb (Rheum.) Plants often contain high stilbene glycosides, such as Turkey rhubarb roots glycoside content Loop rhubarb (Rheum hotaoense CYCheng et CT Kao) up to 3 % (Lu et Dragon, HPLC method for determination of the Loop cultivated rhubarb rhubarb Middle measured glycosides, Journal of pharmaceutical analysis, 2007,27: ll), Rhapontinum cultivated in parts Rheum tanguticum (Rhe coffee tangutic coffee Maxim, ex . BALF) rhizome content may reach 10% (Zhang et al., natural product Research and Development, 2005, 17: 217).

[OOO9] 本发明人在对野生和种植的拉萨大黄(Rheum lhasaense AJLi et PK Haiao) 根茎进行系统的成分研究中首次发现该植物含有丰富的芪苷类成分,除去氧土大黄苷、虎杖苷外(中国发明专利200510010757. X, 200710066456. 8),还含有3〜5%的曲札芪苷。 [OOO9] The present invention is carried out constituents of the system of wild and cultivated la rhubarb (Rheum lhasaense AJLi et PK Haiao) rhizomes first discovered the plant rich in stilbene glycosides, removing oxygen rhaponticin, polydatin outer (Chinese patent 200510010757. X, 200710066456. 8), further comprising a curved sheaf ~ 5% stilbene glycosides. 我们在系统研究拉萨大黄化学成分和制备曲札芪苷提取物、纯品的过程中还发现,与上述富含芪苷成分的大黄植物品种相比,拉萨大黄除含高含量的多羟基芪苷类化合物的特点外, 几乎不含具有潜在毒副作用的蒽醌及蒽醌苷类化学成分,此特点为该植物中芪苷类有效部位或单一成分的开发、产业化生产带来了实用性和应用优势。 We studied the chemical composition and the preparation of la Rheum stilbene glycoside extract music sheaf system, purifying the product also found that compared with the above-described plant varieties of rhubarb stilbene glucoside enriched ingredients, except Rheum Lhasaense POLYHYDROXYSTIBENE glycosides containing high levels of anthraquinone and anthraquinone glycosides outer characteristics of chemical constituents based compound, having a potential almost no side effects, the development of this feature for plants stilbene glycosides active site or a single component, industrial production and brings practicality application advantages.

发明内容 SUMMARY

[0010] 本发明的目的在于克服上述现有技术不足,发明一种曲札芪苷在制备预防和治疗缺血性心脑血管疾病制剂中的应用,及发明一个制备曲札芪苷及含有曲札芪苷提取物的方法。 [0010] The object of the present invention is to overcome the disadvantages of the prior art described above, an inventive song sheaf application stilbene glycosides formulation for the prevention and treatment of ischemic cardiovascular and cerebrovascular diseases, and the invention is a preparation containing the song and song sheaf stilbene glycosides the method of extraction was sheaf stilbene glycosides.

[0011] 本发明是这样实现的: [0011] The present invention is implemented as follows:

[0012] 曲札芪苷在制备预防和治疗缺血性心脑血管疾病制剂中的应用,所述曲札芪苷为化合物(E)-l-(3,5-二羟苯基)-2-(3-羟基-4-0-eD-吡喃葡萄糖苯基)乙烯,化学结构 [0012] Application of curvature sheaf stilbene glycosides formulation for the prevention and treatment of ischemic cardiovascular disease, said curved Sapporo stilbene glucoside compound (E) -l- (3,5- dihydroxyphenyl) -2 - (3-hydroxy-phenyl -4-0-eD- glucopyranosyl) ethylene, the chemical structure

是: [0013] Is: [0013]

<formula>formula see original document page 5</formula> <Formula> formula see original document page 5 </ formula>

[0014] 使用的曲札芪苷为曲札芪苷提取物。 [0014] Qu sheaf stilbene glycosides used for the song sheaf stilbene glucoside extract.

[0015] 所述曲札芪苷提取物,按重量百分比计,曲札芪苷提取物中含有曲札芪苷的有效 [0015] The curved sheaf stilbene glucoside extract, by weight percent, Sapporo curved stilbene glycoside extract contains an effective curvature sheaf stilbene glycosides

成份为50% -100%。 Ingredients 50% to 100%.

[0016] 所述制备方法包括下列步骤: [0016] The preparation method comprises the steps of:

[0017] (1)前处理:前处理步骤包括拉萨大黄原植物根或根茎的采集、净制和/或干燥、 切碎或粉碎; [0017] (1) Pretreatment: pre-processing step includes acquiring the original la rhubarb roots or rhizomes of the plant, the net system and / or dried, pulverized or chopped;

[0018] (2)提取:所述提取是采用水、含水或不含水有机溶剂作为提取溶剂对步骤(1)所述拉萨大黄根或根茎进行提取获得拉萨大黄提取液;所述有机溶剂包括含有广4个碳原子的醇、酮、醚以及它们的混合物。 [0018] (2) Extraction: is the extraction with water, aqueous or non-aqueous organic solvent as an extraction solvent in step (1) the la rhubarb roots or rhizomes are extracted to obtain la rhubarb extract; comprising the organic solvent containing alcohols wide to 4 carbon atoms, ketones, ethers, and mixtures thereof.

[0019] (3)后处理:后处理步骤包括回收或部分回收步骤(2)所得提取液中的有机溶剂 [0019] (3) Processing: or post-treatment step comprising recovering part of the recovery step (2) the resulting organic solvent extract

得到提取浓縮液,所得提取浓縮液可加入或不加水,继而将提取液进行静置沉淀,固液分离 Resulting extract concentrate, the resulting extract concentrate may be added or without addition of water, the extract was then left to settle, the solid-liquid separation

除去部分杂质;除杂质后的液相经进一步吸附树脂层析纯化、分段洗脱、洗脱物浓縮析晶获 Removing a portion of the impurities; other liquid impurities after further purified by chromatography on adsorption resin, segment elution eligible for crystallization was concentrated

得到以重量百分比计,含有曲札芪苷为50% _100%的曲札芪苷提取物。 Resulting in weight percent, comprising curved stilbene glycosides sheaf 50% _100% sheaf curved stilbene glucoside extract.

[0020] 上述曲札芪苷提取物的一种制备方法,其步骤(2)所述有机溶剂为甲醇、乙醇、丙 [0020] A method for preparing the above-described curved sheaf stilbene glycoside extract which step (2) said organic solvent is methanol, ethanol, propanol

酮、正丁醇或乙醚;其步骤(2)所述提取溶剂为50〜100%的乙醇或甲醇水溶液。 Ketone, n-butanol or ether; which step (2) the extraction solvent is 50~100% aqueous ethanol or methanol.

[0021] 上述曲札芪苷纯品的一种制备方法,其特征在于,所述制备方法是采用拉萨大黄 [0021] A method for preparing the above-described curved sheaf stilbene glycosides pure product, characterized in that the preparation method employed Rheum Lhasaense

作为原料,通过提取纯化步骤获得所需曲札芪苷纯品;所述曲札芪苷纯品纯度以重量百分 As a starting material, to give the desired product pure music sheaf stilbene glycosides by extraction and purification steps; stilbene glucoside said curved Sapporo weight percent purity of the pure

比计,为90% -100%。 Ratio, 90% -100%.

[0022] 所述曲札芪苷提取物的一种制备方法,其特征在于曲札芪苷提取物是从野生或栽培的拉萨大黄(Rheum lhasaense AJ Li et PK Haiao)的根或根茎中提取。 The [0022] A method for producing curved sheaf stilbene glycoside extract, wherein the curved sheaf stilbene glycoside extract from wild or cultivated la rhubarb (Rheum lhasaense AJ Li et PK Haiao) extracting the root or rhizome. [0023] 曲札芪苷在制备曲札芪苷纯品中的用途。 [0023] Qu Qu sheaf stilbene glycosides prepared sheaf stilbene glycosides use of pure.

[0024] 曲札芪苷提取物在制备预防或治疗心脑血管疾病制剂中的用途。 [0024] Qu sheaf stilbene glycoside extract for the prevention or treatment of cardiovascular and cerebrovascular diseases formulation.

[0025] 曲札芪苷制剂作为各种医药上可接受的制剂或保健食品,可用于制作药物或保健食品。 [0025] Qu sheaf stilbene glycosides of various formulations as a pharmaceutically acceptable formulation or health food, can be used to produce a pharmaceutical or nutraceutical. [0026] 提供曲札芪苷的在制备防治心脑缺血疾病制剂中的应用,所述曲札芪苷为(E)-l-(3,5-二羟苯基)-2-(3-羟基-4-0-eD-吡喃葡萄糖苯基)乙烯或3,5,3', 4'-四羟基芪心脑_3' -OP-葡萄糖苷,曲札芪苷的化学结构式如下。 [0026] In the preparation of disease control formulation provide myocardial and cerebral ischemia sheaf stilbene glycosides song in the song Sapporo stilbene glucoside (E) -l- (3,5- dihydroxyphenyl) -2- (3 - glucopyranose -4-0-eD- hydroxy phenyl) ethylene or 3,5,3 ', 4'-tetrahydroxy-stilbene heart and brain _3' -OP- glucoside, curved sheaf of formula stilbene glycosides follows. [0027] [0027]

HO OH HO OH

/3 2 / 32

HO HO

[0028]目前研究较多的多羟基芪苷类化合物如虎杖苷、土大黄苷、去氧土大黄苷等,其葡萄糖基取代在所示曲札芪苷的化学结构的C-3位,曲札芪苷的糖基则取代在C-3'位,这种类型的芪苷类化合物在抗心脑缺血活性方面目前尚无研究。 [0028] The present study more stilbene glycosides of polyhydroxy compounds such as Polydatin, rhaponticin, deoxy rhaponticin the like, their glucose group substituted at C-3 position of the curve shown in the chemical structure sheaf stilbene glycosides, Qu Sapporo glycosylated stilbene glycosides are substituted at C-3 'position, no study of this type of stilbene glycosides in the activity of anti-myocardial and cerebral ischemia. 本发明对曲札芪苷进行了抗心脑缺血方面的药理和药效研究,发现曲札芪苷纯品及含有曲札芪苷的提取物在急性心脑缺血疾病的模型中均显示了显著的药理学活性。 The present inventors performed the song sheaf stilbene glycosides pharmacology and efficacy studies of cerebral ischemia anticardiolipin aspect, Sapporo found song and stilbene glycosides containing pure music sheaf stilbene glucoside extracts are shown in the model of acute cerebral ischemia in heart disease significant pharmacological activity. 研究采用两种与人类疾病相近的动物模型,测定多种药效学指标,综合评价两种样品对急性心脑缺血的影响。 Research in two animal models of human disease similar measure multiple pharmacodynamic index, comprehensive evaluation of the impact of the two samples of acute myocardial and cerebral ischemia. 试验结果表明,50、 100mg/kg曲札芪苷提取物和15、30mg/kg纯品腹腔注射能明显降低冠状动脉结扎致心肌缺血大鼠血清CK-MB及LDH的释放,显著縮小心肌梗死范围,显示其对心肌缺血带来的损伤有显著的保护作用。 The results showed that, 50, 100mg / kg song sheaf stilbene glucoside extract and 15,30mg / kg intraperitoneal injection of the pure product could significantly reduce coronary artery ligation-induced myocardial ischemia in rat serum CK-MB and LDH release, significantly reduced myocardial infarct range, show that there is a significant protective effect on myocardial ischemia caused by injury. 50、100mg/kg曲札芪苷提取物和15、30mg/kg还可以明显减少MCA0大鼠脑梗塞范围,明显改善大鼠局灶性脑缺血所致的神经功能程度,表明其对脑缺血的损伤具有显著的保护作用。 50,100mg / kg song sheaf stilbene glucoside extract and 15,30mg / kg also significantly reduced infarct size MCA0 rats, significantly improved neurologic ischemia induced focal brain, indicating that the lack of cerebral damage to the blood has a significant protective effect. 多个剂量的曲札芪苷提取物和纯品均能抑制血小板聚集-肾上腺素所致的小数体内血栓形成。 Doses of the plurality sheaf stilbene glucoside extract and pure inhibited platelet aggregation - epinephrine induced fractional thrombus formation in vivo. 上述试验指标显示,曲札芪苷在防治心脑缺血方面具有潜在的临床应用价值,由此,本发明提供曲札芪苷在制备治疗缺血性心脑血管疾病药物中的用途。 Above test indicators, curved sheaf stilbene glycosides have potential clinical value in the prevention and treatment of ischemic heart and brain, thereby, the present invention provides for the preparation of curvature sheaf stilbene glycosides treatment of ischemic cardiovascular disease medicine. [0029] 所述的制备方法是从拉萨大黄的根和/或根茎提取目标物质,所述拉萨大黄为Rheum lhasaense A. J丄i et PK Haiao。 The method of preparing the [0029] target substance is extracted from the root of Rheum Lhasaense and / or rhizomes of Rheum Lhasaense said Rheum lhasaense A. J Shang i et PK Haiao.

[0030] 本发明进一步提供曲札芪苷和含有曲札芪苷提取物的制备方法,所述制备方法包括下列步骤: [0030] The present invention further provides a curved sheaf stilbene glycosides and methods of preparation comprising curved sheaf stilbene glycoside extract, the manufacturing method comprising the steps of:

[0031] (1)前处理:前处理步骤包括拉萨大黄原植物根或根茎的采集、净制和/或干燥、 切碎或粉碎。 [0031] (1) Pretreatment: pre-processing step includes acquiring the original la rhubarb roots or rhizomes of the plant, the net system and / or dried, pulverized or shredded.

[0032] (2)提取:所述提取是采用含水或不含水有机溶剂作为提取溶剂对步骤(1)所述拉萨大黄根或根茎进行提取获得拉萨大黄提取液;所述有机溶剂包括含有广4个碳原子的醇、酮、醚以及它们的混合物。 [0032] (2) Extraction: is the extraction with an aqueous or non-aqueous organic solvent as an extraction solvent in step (1) for extracting the root or rhizome of rhubarb la la rhubarb extract obtained; containing the organic solvent comprises a wide 4 carbon atoms, alcohols, ketones, ethers, and mixtures thereof.

[0033] (3)后处理:后处理步骤包括回收或部分回收步骤(2)所得提取液中的有机溶剂得到提取浓縮液,所得提取浓縮液可加入或不加水,继而将提取液进行静置沉淀,固液分离除去部分杂质;除杂质后的液相经进一步吸附树脂层析纯化、分段洗脱、洗脱物浓縮析晶可得到权利要求2〜3所述的提取物。 [0033] (3) Post-treatment: After the processing step includes recovery or partial recovery of step (2) the resulting organic extract was concentrated to obtain an extraction solvent, the resulting extract concentrate may be added or without addition of water, the extract solution was then the precipitate was allowed to stand, the solid-liquid separation to remove part of the impurities; other liquid impurities after further purified by chromatography on adsorption resin, segment elution crystallization was concentrated to obtain the extract as claimed in claim 2 or 3.

6[0034] 步骤(1)所述前处理步骤中,所述拉萨大黄是指蓼科植物Rheum lhasaense 6 [0034] Step (1) in the pretreatment step, said Rheum Lhasaense refers Polygonaceae Rheum lhasaense

AJ丄i et PKHaiao。 AJ Shang i et PKHaiao. 所述拉萨大黄可以是野生的,也可以是人工种植的,人工引种栽培 The wild Rheum Lhasaense may be, may be planted, the introduction of artificial cultivation of

大黄的成功可以进一步扩大拉萨大黄的药源或产生更优的变种。 Rhubarb can successfully expand Lhasa rhubarb drug source or produce better varieties. 本发明发现,拉萨大黄根 Finding of the present invention, rhubarb root Lasa

及根茎中存在高含量的曲札芪苷,随采收年限、季节、地点以及其它因素的不同,拉萨大黄 And the presence of high levels of music sheaf stilbene glycosides rhizomes harvested with age, season, location, and various other factors, Rheum Lhasaense

根及根茎所含曲札芪苷的量可达3%〜5% (以干重计),此在植物中非常罕见。 Song contained in an amount of roots and rhizomes sheaf stilbene glycosides up to 3% ~ 5% (by dry weight), in this very rare plants.

[0035] 此外,本发明还惊奇发现,拉萨大黄根和根茎中几乎不含蒽醌类化合物,此在大黄 [0035] Further, the present inventors have also surprisingly found that, in the roots and rhizomes Rheum Lhasaense almost no anthraquinones, this rhubarb

类植物中为罕见,这是拉萨大黄的一项非常重要的特征:其一,应用拉萨大黄入药不存在对 For rare plants, Lhasa rhubarb This is a very important feature: First, the application does not exist to Lhasa rhubarb medicine

蒽醌类化合物潜在严重毒副作用的担忧;其二,应用拉萨大黄提取二苯乙烯类活性成分时, When Second, rhubarb extract applied la stilbene active ingredient; anthraquinones potentially serious toxicity concerns

其提取工艺可以不受蒽醌类化合物特别是蒽醌苷的干扰,因此工艺可以更简便,上述特点 Extracting process can not anthraquinone compounds, especially anthraquinone glycoside interference, and thus the process may be more simple, the above-described characteristics

为拉萨大黄中芪苷类有效部位或单一成分的开发、利用和产业化生产带来了应用优势和实用性。 Lhasa rhubarb for the development of effective parts of the stilbene glycosides or single component, utilization and industrial production brings advantages and practical applications.

[0036] 拉萨大黄地上茎叶中二苯乙烯类成分含量很低,作为提取原料的价值相对较低, 但是仍可作为二苯乙烯类化合物的来源之一。 [0036] The ground leaves and stems of rhubarb la stilbenes component content is very low, as the relatively low value of the extraction of raw materials, but still be used as a source of stilbene compounds. 本发明优选采用拉萨大黄的地下根和根茎作为提取原料,但不排除以其整个植株作为提取原料。 The present invention preferably uses Lasa subterranean rhubarb roots and rhizomes extract as a raw material, but does not preclude its whole plants as the raw material for extraction.

[0037] 采集的拉萨大黄一般需要经过净制、切制和/或粉碎等前处理。 [0037] collected Rheum Lhasaense generally require net system, cut and / or crushed before treatment. 所述净制可包括去除采集过程中附着于拉萨大黄根和根茎上的泥土等杂物,还可以包括将根和根茎与地上茎叶分开等。 The net system acquisition process may include removing la rhubarb roots attached to the roots and clay debris, may further comprise separating the ground roots and rhizomes of stems and leaves. 所采集的拉萨大黄可以应用其鲜品,也可以采用其风干、晒干或机器干燥的干品;所得此根和根茎或整个植株优选进行适当切制或粉碎以利提取。 Rheum Lhasaense acquired fresh goods which may be applied, which may be used dry, dried or machine dried dry matter; this resultant whole plants or roots and rhizomes preferably appropriately cut or pulverized to take Liti.

[0038] 步骤(2)所述有机溶剂为甲醇、乙醇、丙酮、正丁醇或乙醚。 [0038] Step (2) the organic solvent is methanol, ethanol, acetone, diethyl ether, or n-butanol. 其提取溶剂优选乙醇或甲醇的水溶液;其中特别优选乙醇水溶液作为提取溶剂;进一步地,本发明优选采用45%〜95%乙醇水溶液作为提取溶剂。 Extracting solvent, preferably aqueous ethanol or methanol; wherein the aqueous ethanol is particularly preferred as solvent; Further, the present invention preferably employs an aqueous solution of 45% ~ 95% ethanol as the extraction solvent.

[0039] 步骤(2)所得拉萨大黄提取液需经适当的后处理方可获得曲札芪苷提取物。 [0039] Step (2) la rhubarb extract obtained by suitable after-treatments need only obtain curved sheaf stilbene glucoside extract. 由于拉萨大黄所含的一些主要的芪及芪苷类物质如土大黄苷元、去氧土大黄苷元、去氧土大黄苷、虎杖苷水溶性有限,本发明发现,回收提取液中的有机溶剂和/或适当浓縮提取物可以使提取物所含的部分脂溶性芪及芪苷类化合物、杂质从提取液中析出,而曲札芪苷由于水溶性较好则保留在上清液中。 Because some major stilbene and stilbene glycosides contained substances such as soil Rheum Lhasaense aglycone, Rumex deoxy aglycone, deoxy rhaponticin, polydatin limited water-solubility, the present inventors have found that the recovery of the organic extract the solvent and / or a suitable portion of the concentrated extract can make the fat-soluble stilbene and stilbene glycosides contained in the extract, the extract from the precipitation of impurities, but since the song Sapporo stilbene glycosides is preferably soluble in the supernatant is retained . 其转移率可达85%以上。 Its transfer rate of up to 85%. 上清液中的曲札芪苷可以采用合适的有机溶剂萃取进行纯化,例如采用正丁醇、乙酸乙酯_甲醇混合溶剂、正丁醇_ 二氯甲烷混合溶剂等萃取;此类化合物还可以采用连续逆流萃取法等本领域内熟知的方法萃取纯化。 Qu supernatant sheaf stilbene glycosides suitable organic solvent extraction may be purified using, for example with n-butanol, a mixed solvent of ethyl acetate and methanol _, _ extracted with methylene chloride mixed solvent of n-butanol and the like; such compounds may also be extraction purification method well known in the art of continuous countercurrent extraction method.

[0040] 本发明中优选采用大孔吸附树脂层析法纯化上清液中的二苯乙烯类化合物。 [0040] In the present invention, preferred macroporous resin chromatography in the supernatant was purified stilbene compound. 上述后处理步骤中,随着提取液中的有机溶剂的回收,上清液上样于大孔吸附树脂柱时,上清液中所含的曲札芪苷易于被树脂柱床所吸附;上样吸附后,树脂柱床可采用水以及低浓度有机溶剂水溶液洗涤以进一步去除极性较大的杂质,随后可采用较高浓度的有机溶剂水溶液分步洗脱所需的化学成分,所得洗脱溶液经浓縮、析晶、干燥即可获得所述的曲札芪苷提取物。 After the above processing step, the extract recovered with organic solvent, the supernatant was loaded onto a macroporous resin column, the supernatant contained in the song sheaf stilbene glycosides readily adsorbed by the resin bed; the after sample adsorption, the resin bed can be washed with water and an aqueous solution of a low concentration of organic solvent to further remove more polar impurities, followed by a higher concentration of the organic solvent can be an aqueous solution of the desired chemical composition of the elution step, the resulting elution the solution was concentrated, crystallization, drying curve can be obtained glycoside extract sheaf stilbene said. 提取物可采用醇水或单纯的水溶液重结晶进行进一步精制来提高曲札芪苷的纯度,通过反复重结晶、脱色或其它纯化手段可得权利要求7〜8所述的纯品。 Recrystallized from aqueous or aqueous alcohol extract can be further purified to simply increase the purity of the song sheaf stilbene glycosides, by repeated recrystallization, discolored or other purification means available product according to claim 7~8 pure.

[0041] 采用大孔树脂吸附上清液所含曲札芪苷时,柱床体积一般按每100ml吸附树脂(装柱后体积)吸附l〜3g上清液设置,这时一般可以获得良好的吸附转移率。 [0041] When macroporous resin contained in the supernatant using curved sheaf stilbene glycosides, 100ml bed volume adsorption resin per ships (installed after the column volume) of the supernatant was adsorbed l~3g provided, then generally possible to obtain good adsorption transfer rate. 上清液上样吸附后,可采用水溶液、10%〜15%的乙醇溶液或甲醇水溶液洗涤除去大部分水溶性杂质, After adsorption of the supernatant sample, may be used an aqueous solution, washed with aqueous methanol or ethanol solution of 10% ~ 15% of the most water-soluble impurities was removed,

7随后采用约25%〜50%的乙醇或甲醇水溶液即可洗脱吸附于大孔树脂的曲札芪苷。 7 can then adsorbed on a macroporous resin was eluted curved sheaf stilbene glycosides using about 25% ~ 50% aqueous ethanol or methanol. 本发明中,优选的大孔吸附树脂为弱极性和中极性的树脂,例如商品型号D101、D1300、AB8型吸附树脂。 In the present invention, the preferred resin is a macroporous weakly polar and polar resins, such as Item No. D101, D1300, AB8 adsorption resin. 在合适的条件下,大孔树脂吸附法所得产物中,曲札芪苷总含量可达约50%至85%,转移率为80%〜95%。 Under suitable conditions, the resulting macroporous adsorption resin product, the total content of the song Sapporo stilbene glycosides up to about 50-85%, the transfer ratio of 80 ~ 95%.

[0042] 经过上述纯化步骤后,曲札芪苷已经得到富集,采用这些提取物作为原料可以方便地制备曲札芪苷含量更高的提取物和纯品。 [0042] After these purification steps, the song has been Sapporo stilbene glucoside enriched using these extracts can be conveniently prepared stilbene glucoside content of greater curvature sheaf extract and pure as the starting material. 例如,本发明中,采用大孔树脂吸附法所得洗脱部分为原料,通过合适浓縮,析晶可得含量为70%〜89%的曲札芪苷提取物,此提取物采用反复重结晶、活性碳脱色、硅胶层析、聚酰胺等层析结合重结晶法纯化可获得纯度高达90%以上的曲札芪苷纯品。 For example, the present invention, a macroporous resin adsorption resulting eluted fractions as a raw material, by a suitable concentration, crystallization available content of 70% ~89% of curvature sheaf stilbene glucoside extract, the extract is recrystallized repeatedly , activated carbon decolorization chromatographed on silica gel, polyamide, etc. Chromatograph recrystallization purification, the purity of more than 90% pure song sheaf stilbene glycosides.

[0043] 通过上述步骤制备的提取物是以曲札芪苷为主要化学成分的混合物。 [0043] The extract is prepared by the above step song sheaf stilbene glucoside mixture main chemical constituents. 可以理解,此提取物中曲札芪苷的含量随原料批次不同、处理工艺参数变化而有所变化。 It will be appreciated, this extract song sheaf stilbene glucoside content with different batches of raw materials, process parameters variations vary. 一般地,按重量百分比计,所述提取物中含有约50%〜85%的曲札芪苷。 Generally, by weight percent, the extract contains about 50% ~ 85% of the curved sheaf stilbene glycosides.

[0044] 显然,上述提取物可以用于进一步精制纯化以获得曲札芪苷纯品。 [0044] Obviously, the above extract may be further purified for purification to obtain pure music sheaf stilbene glycosides. 本发明中,所谓纯品是指化合物的纯度不低于90%。 In the present invention, the term pure refers to purity of the compound is not less than 90%.

[0045] 由此本发明进一步提供拉萨大黄提取物在制备曲札芪苷纯品,以及曲札芪苷提取物在制备该曲札芪苷纯品中的用途。 [0045] Thus the present invention further provides a la rhubarb extract in preparing pure music sheaf stilbene glycosides and glycoside extract song Sapporo stilbene use of the song sheaf stilbene glucoside in the preparation of pure product.

[0046] 研究结果显示,曲札芪苷提取物及曲札芪苷纯品在作为制备治疗心脑血管疾病的 [0046] The results show that, Qu sheaf stilbene glucoside extracts and pure music sheaf stilbene glycosides prepared as in the treatment of cardiovascular and cerebrovascular diseases

治疗药物方面有广泛的用途。 Drug treatment of a wide range of uses. 所述的有效部位中活性成分的结构清楚、组成明确,有利于各 The effective part of the apparent structure of the active ingredient, a clear composition, the benefit of all

级产品的质量监测和控制,符合天然产物复合制剂的发展方向和要求。 Quality monitoring and control stage product line with the development of natural products and complex formulation requirements. 另外,本发明所述有 Further, the present invention has

效部位的制备方法简单易行,生产成本低,可连续操作,具有极强的实用性。 Simple and efficient method of preparing the site, low production cost, can be operated continuously, have highly practical.

[0047] 按照常规的制剂工艺,可以将本发明之提取物制备成任何一种适用于临床应用的 [0047] According to the conventional preparation process, the extract can be prepared into any of the present invention suitable for use in clinical applications

药物制剂。 Pharmaceutical preparations. 如片剂,胶囊剂,丸剂,散剂,膏剂,分散剂,颗粒剂,粉针剂,水针剂等。 Such as tablets, capsules, pills, powders, pastes, dispersants, granules, powder, water injection and the like.

[0048] 同样,本发明之曲札芪苷提取物也可以与其它活性成分联合使用形成新的制剂。 [0048] Similarly, the present invention sheaf song stilbene glucoside extracts may be used in combination to form a new formulation with other active ingredients.

显然,这些含有本发明所述有效部位的制剂在本发明的权利要求范围内。 Clearly, these preparations contain the active site of the present invention as claimed in the present invention within the scope of the claims.

附图说明 BRIEF DESCRIPTION

[0049] 图1为本发明薄层鉴别(碘显色)。 [0049] FIG TLC (iodine color development) of the present invention.

[0050] 图2野生拉萨大黄原料供试品液HPLC典型图谱(曲札芪苷,RT :7. 862)。 [0050] FIG. 2 wild Rheum Lhasaense test material was typical HPLC profiles (Triton sheaf stilbene glycosides, RT:. 7 862). [0051] 图3种植拉萨大黄原料供试品液HPLC典型图谱(曲札芪苷,RT :7. 885)。 [0051] FIG. 3 la planting raw rhubarb liquid test typical HPLC profiles (Triton sheaf stilbene glycosides, RT:. 7 885). [0052] 图4曲札芪苷提取物供试品液HPLC典型图谱(曲札芪苷,RT :7. 878)。 [0052] FIG. 4 sheaf stilbene glycoside extract test was typical HPLC profiles (Triton sheaf stilbene glycosides, RT:. 7 878). [0053] 图5曲札芪苷纯品HPLC典型图谱(曲札芪苷,RT :7. 896)。 [0053] FIG. 5 sheaf stilbene glucoside pure typical HPLC profiles (Triton sheaf stilbene glycosides, RT:. 7 896).

[0054] 其中在图l中:l.表示去氧土大黄苷对照品液;2.表示曲札芪苷对照品液;3.表示野生拉萨大黄原料(20080913)供试品液;4.表示种植拉萨大黄原料(20080915)供试品液;5.表示种植拉萨大黄原料(20080915)供试品液;6.表示种植拉萨大黄原料(20080915)供试品液。 . [0054] In Figure l wherein: l represents deoxy rhaponticin reference substance solution; 2 represents a curve sheaf stilbene glycosides reference substance solution; 3 represents wild Rheum Lhasaense material (20,080,913) the test fluid; represents 4.. Rheum Lhasaense planting material (20,080,915) test solution; 5 represents la Rheum planting material (20,080,915) the test liquid; 6 represents Rheum Lhasaense planting material (20,080,915) the test liquid.

具体实施方案 Specific embodiments

[0055] 以下通过具体实施例将进一步说明本发明的内容。 [0055] The following will further illustrate the present invention by way of specific examples. 下文实施例仅为就本发明所述有效部位的来源、制备方法、化学组成及其用途的特殊说明,显然,本发明权利范围不受这些实施例限制。 Example Hereinafter will only source embodiment of the present invention the active parts of the production method, the chemical composition described and its special-purpose, clearly, the scope of the claimed invention is not limited by these examples.

[0056] 实施例1 :拉萨大黄原料及曲札芪苷提取物中曲札芪苷的定性鉴别与定量检测[0057] (1)样品来源 [0056] Example 1: Qualitative and quantitative identification detection curved Sapporo stilbene glycosides [0057] (1) Materials and song Rheum Lhasaense sheaf stilbene glycoside extract sample origin

[0058] 野生拉萨大黄根茎:收购于西藏拉萨地区,除去杂质,洗净,切成片或块,晒干。 [0058] Rhubarb Rhizome wild Lasa: Acquisition of Lhasa area, remove impurities, washed, cut into pieces or blocks, and dried. 批号:20080913 Lot: 20080913

[0059] 种植拉萨大黄根茎:收获于云南香格里拉县,除去杂质,洗净,切成片或块,晒干。 [0059] Growing roots of Rheum Lasa: La County Yunnan to harvest, remove impurities, washed, cut into pieces or blocks, and dried. 批号:20080915 Lot: 20080915

[0060] 对照品:曲札芪苷,自制,纯度99%,批号:080602。 [0060] Reference Sample: Qu Sapporo stilbene glycosides, made, purity 99%, batch number: 080602. [0061] (2)样品及标准品溶液配制 [0061] (2) Sample preparation and standard solution

[0062] 对照品溶液配制:精密称取以五氧化二磷真空干燥的曲札芪苷对照品适量,以甲醇配成lmg/ml对照品储备液,27%甲醇_3%乙酸溶液稀释成25g/ml作为标准品溶液。 [0062] The control solution preparation: Accurately weighed dried curved sheaf stilbene glycosides controlled amount of phosphorus pentoxide in vacuo, methanol dubbed lmg / ml reference standard stock solution, 27% methanol solution diluted to _3% acetic acid 25g / ml as a standard solution. [0063] 拉萨大黄原料供试品溶液制备:精密称取以五氧化二磷真空干燥的上述拉萨大黄根茎粉末10g,95X乙醇回流提取3次(120min/次),合并提取液,滤纸过滤,滤液进一步以0. 45 ii m微孔滤膜过滤,弃初滤液,取续滤液作为拉萨大黄供试品溶液。 [0063] Preparation of the test solution Rheum Lhasaense material: phosphorus pentoxide accurately weighed to the dried rhizome of rhubarb powder 10g la vacuo, 95X ethanol was extracted 3 times (120min / times), the combined extracts, filter paper, the filtrate further to 0. 45 ii m filter membrane, primary filtrate discarded, the filtrate as the test solution Rheum Lhasaense. [0064] 曲札芪苷提取物供试品溶液制备:精密称取以五氧化二磷真空干燥的实施例2所述曲札芪苷提取物适量,以甲醇配成10mg/ml样品储备液,精密吸取lml,置于25ml容量瓶中,27 %甲醇定容,密塞,摇匀,过0. 45 ii m微孔滤膜,弃初滤液,取续滤液作为拉萨大黄提取物供试品溶液。 [0064] Qu sheaf stilbene glycosides extract was prepared for the test solution: accurately weighed in a dry sheaf Example 2 stilbene glucoside extracts amount of phosphorus pentoxide in vacuo, methanol dubbed 10mg / ml stock solution of the sample, lml of precision drawing, placed in 25ml volumetric flask, volume 27% methanol, Mesa, shake, through 0. 45 ii m microporous membrane, primary filtrate discarded, the filtrate la rhubarb extracts as test solution .

[0065] (3)定性鉴别:薄层色谱法检查 [0065] (3) qualitative identification: TLC inspection

[0066] 本法采用硅胶薄层层析。 [0066] Act using silica gel thin layer chromatography. 取上述供试品液及对照品液各41,分别点于同一硅胶薄 Take the above test solution and reference solution in each 41, point in the same silica gel thin

层层析板(硅胶G)板上,以氯仿:甲醇(体积比,7.5 : 2.5)为展开剂展开,取出,晾干,置 Layer chromatography plate (silica gel G) plates with chloroform: methanol (volume ratio, 7.5: 2.5) as developing solvent, removed, dried, set

于碘缸中显色,在对照品相同的位置,供试品应有相应的斑点。 Iodine color in the cylinder, at the same reference position, the test should have the appropriate spot.

[0067] (4)供试品所含曲札芪苷含量定量检测:高效液相色谱法(HPLC) [0067] (4) contained in the test song sheaf stilbene quantitative glucosinolate content: High Performance Liquid Chromatography (HPLC)

[0068] 色谱条件:采用Zorbax SB-C18 (4. 5 X 150mm, Agilent)反相色谱柱,27 %甲 [0068] Chromatographic conditions: A Zorbax SB-C18 (4. 5 X 150mm, Agilent) reverse-phase column, 27% A

醇_3%醋酸水为流动相,流速0. 8ml/min,用DAD紫外检测仪进行检测,检测波长319nm。 Alcohol _3% aqueous acetic acid as the mobile phase, a flow rate of 0. 8ml / min, detection with UV detector DAD, detection wavelength 319nm.

[0069] 标准曲线建立:上述对照品储备液分别稀释成5、15、25、35、45ii g/ml溶液,各取 [0069] The standard curve established: above the reference stock solution were diluted 5,15,25,35,45ii g / ml solution, depicting

10 iU分别注入液相色谱仪,按上述色谱条件测定峰面积,以各对照品峰面积(Y,A)对进样 10 iU were injected into the liquid chromatograph, the peak area was measured by the above chromatographic conditions, the peak area of ​​each reference standard (Y, A) of the sample

量(X,mg)进行线性回归,回归方程:Y = 6. 3324X+0. 1158,r = 0. 9999 (曲札芪苷,n = 5)。 Amount (X, mg) linear regression, the regression equation: Y = 6. 3324X + 0 1158, r = 0. 9999 (Sapporo curved stilbene glycosides, n = 5)..

[0070] 样品测定:上述拉萨大黄原料供试品溶液、曲札芪苷提取物供试品溶液各10 iU [0070] Sample measurement: the above raw rhubarb la test solution, extract song Sapporo stilbene glycoside each test solution 10 iU

分别注入液相色谱仪中,根据标准曲线计算拉萨大黄原料提取物、曲札芪苷提取物中曲札 Were injected into the liquid chromatograph, the computing la raw rhubarb extract standard curve, curved Sapporo stilbene glycoside extract song sheaf

芪苷的含量。 Stilbene glucoside content.

[0071] (4)结果: [0071] (4) Results:

[0072] i)定性鉴别:拉萨大黄原料提取物及曲札芪苷提取物的典型薄层色谱图如图3所示,结果表明,在对照品相同的位置,供试品均有相应的斑点。 [0072] i) qualitative identification: TLC typical raw rhubarb extract FIG la and song sheaf stilbene glucoside extracts shown in Figure 3, the results show that the same reference position, the test has a corresponding spot .

[0073] ii)曲札芪苷含量检测:拉萨大黄原料及其曲札芪苷提取物的典型HPLC谱图如图4〜6所示,结果表明,拉萨大黄野生药材原料和种植药材原料中曲札芪苷的含量分别为4. 38%和4. 18%,提取物中曲札芪苷的含量为78.5%。 [0073] ii) stilbene glycosides sheaf song content detection: Typical HPLC la Rheum stilbene glucoside starting material and curved sheaf extract spectrum shown in FIG. 4 ~ 6, the results showed that wild herbs la rhubarb medicinal raw materials and planting Qu Sapporo stilbene glucoside content of 4.38% and 4.18%, respectively, the content of the extract song sheaf stilbene glycosides of 78.5%. [0074] 实施例2 :曲札芪苷提取物制备 [0074] Example 2: Preparation of curvature sheaf stilbene glycosides extracted

[0075] (1)材料:拉萨大黄根茎:收购于西藏拉萨地区,除去杂质,洗净,切成片或块,晒干。 [0075] (1) Material: Lasa rhubarb roots: Acquisition of Lhasa area, remove impurities, washed, cut into pieces or blocks, and dried. 批号:20080913 Lot: 20080913

[0076] 树脂:DIOI大孔吸附树脂,天津市海光化工有限公司产品,按照生产厂说明书进 [0076] Resin: DIOI macroporous resin, sea-Chemical Co., Ltd, Tianjin, according to the manufacturer instructions into

行预处理。 Pretreated.

[0077] (2)制备 [0077] (2) Preparation of

[0078] i)拉萨大黄前处理及其曲札芪苷含量测定: [0078] i) pre-processing and music Rheum Lhasaense sheaf stilbene glycosides Determination:

[0079] 采集的拉萨大黄植株,去泥晾干,分离地下根茎和地上茎叶,粉碎机分别将其粉碎成粗粉末。 [0079] Rheum Lhasaense plants collected, dried mud to isolated ground rhizomes and stem and leaf, respectively mill pulverized into a coarse powder.

[0080] 取地下根茎和地上茎叶粉末各100g,用95%乙醇浸泡回流提取,120分钟/次,间隔取样进行HPLC检测,直至提取液中基本不含曲札芪苷,合并提取液,按实施例1中所述方法方法曲札芪苷的含量,此检测结果作为药材所含该化合物总量。 [0080] Take ground rhizomes and stem and leaf of each powder 100g, soaked with 95% ethanol extraction at reflux for 120 minutes / time, sampled for HPLC detection intervals until the mixture is substantially free of stilbene glycosides sheaf song, the combined extracts extracted, by song content Sapporo stilbene glycoside in Example 1 embodiment of the methods, the detection result of the total amount of compounds contained as a medicine.

[0081] 结果:批号20080913的拉萨大黄地下根茎和地上茎叶中的所含曲札芪苷化合物总量分别为5. 01 %和0. 001%。 [0081] Results: total curvature stilbene glucoside compounds contained sheaf lot 20080913 and rhizomes of Rheum Lhasaense ground stems and leaves was 5.01% and 0.001%, respectively. [0082] ii)提取: [0082] ii) Extraction:

[0083] 拉萨大黄地下根茎粉末每份600g,分别以50%、75%、95%乙醇;50%、90%甲醇; [0083] la rhizome of rhubarb powder per 600 g of, respectively, 50%, 75%, 95% ethanol; 50%, 90% methanol;

50%丙酮;正丁醇作为提取溶剂提取。 50% acetone; n-butanol extraction as an extraction solvent. 各份药材分别以上述不同提取溶剂共5. 4L回流提取 In each of these different parts of herbs were co-extraction solvent refluxing 5. 4L

3次(2. 4L/1. 8L/1. 2L) , 120min/次。 3 (2. 4L / 1. 8L / 1. 2L), 120min / times. 分别合并提取液,HPLC法检测提取液中曲札芪苷平均 Extracts were combined, HPLC assay extract stilbene glycosides average curvature sheaf

量(A):,计算曲札芪苷转移率(B):提取液中曲札芪苷总量/药材曲札芪苷总量X100%。 The amount of (A) :, TORTUOSITY sheaf stilbene glycosides transfer rate (B): total extract song sheaf stilbene glycosides / total curved sheaf stilbene glycosides medicine X100%.

[0084] 提取试验结果如表1所示。 [0084] Extraction test results are shown in Table 1.

[0085] 表1 :不同提取溶剂的提取试验结果 [0085] Table 1: Test results of different extraction solvents to extract the

[0086] [0086]

提取液 50%乙醇 75%乙醇 95%乙醇 50%甲醇 90%甲醇 50%丙酮 正丁醇 Extract 50% ethanol 75% ethanol 95% ethanol 50% methanol and 90% methanol 50% acetone-butanol

A 22.50g 23.10 g 27.12 g 21. 60 g 28.68 g 27. 18 g 24. 36 g A 22.50g 23.10 g 27.12 g 21. 60 g 28.68 g 27. 18 g 24. 36 g

B 74.8 % 76.8% 90.2 % 70.6 % 93.5 % 90.4 % 81.2 % B 74.8% 76.8% 90.2% 70.6% 93.5% 90.4% 81.2%

[0087] A :HPLC法检测的提取液中曲札芪苷化合物量;B :曲札芪苷转移率 [0087] A: the amount of the compound sheaf stilbene glycoside extract was detected by HPLC Qu; B: Transfer rate QU sheaf stilbene glycosides

[0088] 上述提取试验中,以50%、75%、95%乙醇;50%、90%甲醇;50%丙酮;正丁醇作 [0088] The extraction experiment, 50%, 75%, 95% ethanol; 50%, 90% methanol; 50% acetone; n-butanol as

为提取溶剂提取时,提取步骤中曲札芪苷转移率在70%〜95%之间。 When the extraction is the extraction solvent, the extraction step Qu sheaf stilbene glycosides transfer rate of 70 ~ 95%. 可见它们均可作为该 They can be seen as the

化合物的有效提取溶剂,其中,鉴于乙醇的安全、廉价且提取转移率较高,因此本发明优选 Efficient extraction solvent for the compound, wherein, in view of safety ethanol extraction inexpensive and high transfer rate, the present invention is therefore preferably

乙醇水溶液作为提取溶剂。 Aqueous ethanol as solvent.

[0089] iii)后处理: [0089] iii) Post-treatment:

[0090] 沉淀处理:分别合并步骤ii)所得各提取溶液,减压(0. 1〜0. 08Mpa,6(TC )回收其 [0090] The precipitation process: each merging step ii) each of the resulting extraction solution under reduced pressure (0. 1~0 08Mpa, 6 (TC.) Recover its

中所含的有机溶剂,加水定容至600mL后,混合均匀,4"放置24h,离心(4000rpmX 30min), After the organic solvent contained, add water to 600 mL, mixed, 4 "Place 24h, centrifuged (4000rpmX 30min),

得上清液和沉淀。 To obtain supernatant and a precipitate. 沉淀真空干燥,称重(C) ;HPLC法检测上清液和沉淀中曲札芪苷的量(D、 The precipitate was dried in vacuo, weighed (C); HPLC method detects the amount of supernatant and pellet song sheaf stilbene glycosides (D,

)。 ). 计算上清液和沉淀中曲札芪苷转移率(F、G):上清液或沉淀曲札芪苷总量/提取液曲 Qu calculated supernatant and pellet sheaf stilbene glycosides transfer rate (F, G): the total amount of the supernatant or precipitate sheaf stilbene glycosides curved / bent extract

札芪苷总量X100%。 Sapporo total stilbene glycosides X100%.

[0091] 表2 :沉淀处理试验结果 [0091] Table 2: Test results precipitating

[0092]<table>table see original document page 11</column></row> <table> [0092] <table> table see original document page 11 </ column> </ row> <table>

[0093] C:沉淀量D:沉淀中曲札芪苷量[0094] E :上清液中曲札芪苷量E :沉淀中曲札芪苷转移率 [0093] C: amount of precipitate D: precipitation amount Qu sheaf stilbene glycosides [0094] E: supernatant sheaf stilbene glycosides quantity Qu E: Qu precipitate sheaf stilbene glycosides Transfer Rate

[0095] F :上清液中曲札芪苷转移率; [0095] F: Supernatant song sheaf stilbene glycosides transfer rate;

[0096] 表2数据显示,上述各组提取液浓縮除有机溶剂加水后低温静置,除正丁醇组外,其余各组样品均产生沉淀,沉淀中曲札芪苷的转移率在2%〜5%之间,沉淀量在10〜30g之间,上清液中曲札芪苷转移率在85%〜95%之间,可见该步骤处理可有效去除脂溶性杂质,且曲札芪苷的损失较小。 [0096] Data in Table 2 show, each of the groups except the low temperature concentrated extract was allowed to stand after addition of water the organic solvent, n-butanol addition to the group, the other set of samples were each precipitation, the precipitation rate of the transfer curve sheaf stilbene glycosides 2 % ~ 5% between, the amount of precipitation between 10~30g, stilbene glycosides sheaf supernatant curved transfer rate of 85 ~ 95%, showing that the fat-soluble step process can effectively remove impurities, and stilbene curved Sapporo glycosides smaller losses. 因此本发明把该步骤作为制备曲札芪苷提取物的一个重要环节。 Thus, the present invention is the important part of this step as a sheaf prepare triamcinolone stilbene glucoside extract. [0097] 上清液处理:除正丁醇提取液外,合并其它全部上清液,再次过滤,HPLC法检测所得滤液中的曲札芪苷总量,将其平均分成4份,分别上样于4根D101大孔吸附树脂柱(小5. Ocm, 105cm,柱床体积2000ml)和HPLC检查D101树脂柱上样流出液,未见二苯乙烯类化合物泄漏。 [0097] The supernatant process: in addition to butanol extract, but all of the other combined supernatants were filtered again, the total amount of the resulting curve Sapporo stilbene glycosides detected by HPLC the filtrate, which were divided into 4 parts, respectively, the sample 4 to D101 macroporous resin column (small 5. Ocm, 105cm, bed volume 2000ml) and checked by HPLC D101 resin column effluent sample, no leakage stilbene compound. 4根D101树脂柱分别顺次以1000ml水、1000ml 15%乙醇或10%甲醇洗涤,HPLC检查洗涤流出液,流出液基本不含或仅含少量曲札芪苷。 4 resin column D101 to 1000ml were sequentially washed with water, 1000ml 15% ethanol or 10% methanol, HPLC and check the wash effluent, the effluent is substantially free of curvature or only small amounts of stilbene glycosides sheaf. D101树脂柱再分别以25、50 % 7醇或20 %甲醇、45 %甲醇洗脱,收集洗脱液,减压浓縮,真空干燥得曲札芪苷提取物,计量所得终产物的重量(H),HPLC法检测终产物中曲札芪苷总量(I);计算终产物中的曲札 D101 resin column 25, 50, respectively, then 7% alcohol or 20% methanol, 45% methanol, the eluate was collected and concentrated under reduced pressure, and dried in vacuo to give song sheaf stilbene glucoside extract, the resulting measurement of the final product weight ( H), HPLC was used to detect the total amount of (I) final product Qu sheaf stilbene glycosides; TORTUOSITY sheaf final product

芪苷的含量(J):终产物中曲札芪苷总量/终产物重量X100X;计算曲札芪苷转移率(K): Stilbene glucoside content of (J): the end product of the total curvature sheaf stilbene glycosides / weight of the final product X100X; TORTUOSITY Sapporo stilbene glycosides transfer rate (K):

终产物中曲札芪苷总量/上样滤液中曲札芪苷总量X 100% 。 Qu sheaf total stilbene glycosides final product / total curved loading sheaf stilbene glycosides filtrate X 100%. [0098] 表3上清液处理试验结果 [0098] Table 3 Test results of the processing supernatant

树脂柱D101大孔吸附树脂 D101 macroporous resin column, the resin

[0099] [0099]

<table>table see original document page 11</column></row> <table> <Table> table see original document page 11 </ column> </ row> <table>

[0100] H :终产物量; I :终产物量中曲札芪苷总量 [0100] H: the amount of the final product; I: Sapporo stilbene glycosides total amount of the final product Qu

[0101] J:终产物量中曲札芪苷含量K:曲札芪苷转移率; [0101] J: the amount of the final product sheaf music content stilbene glycosides K: Qu sheaf stilbene glycosides transfer rate;

[0102] 每根树脂柱上样滤液(750ml)中曲札芪苷总量:33. 82g[0103] 表3数据显示,从吸附树脂上洗脱部分的曲札芪苷含量在50%〜85%之间,曲札芪 [0102] Each sample filtrate resin column (750ml) stilbene glycosides total sheaf Qu:. 33 82g [0103] Data in Table 3 show that the adsorption resin is eluted from the curved portion of the sheaf stilbene glycoside content of 50% ~ 85 % between Qu Qi Sapporo

苷的转移率在85%〜96%之间。 Glycosides transfer rate between 85% ~96%. 其中,鉴于乙醇的安全、廉价,因此本发明优选乙醇水溶液 Wherein, in view of the ethanol is safe, inexpensive, and therefore the present invention is preferably aqueous ethanol

作为洗脱剂,洗脱浓度优选25%〜50%之间,可以预见,随着洗脱液乙醇浓度的降低,洗脱 As eluant, eluting with 25% ~ 50% concentration is preferably, can be expected, with decreasing the ethanol concentration of the eluate, eluted

部分曲札芪苷的含量会有所上升,而转移率有所下降,乙醇浓度升高则反之。 Song content sheaf stilbene glycosides portion will rise, while the transfer rate is decreased, the ethanol concentration is increased and vice versa.

[0104] 实施例3 :由拉萨大黄提取物及曲札芪苷提取物制备纯品曲札芪苷 [0104] Example 3: Preparation of pure product were extracted by curve sheaf stilbene glycosides la rhubarb extracts and stilbene glycosides song sheaf

[0105] (1)材料:拉萨大黄根茎:收购于西藏拉萨地区,除去杂质,洗净,切成片或块,晒 [0105] (1) Material: Lasa rhubarb roots: Acquisition of Lhasa area, remove impurities, washed, cut into pieces or blocks, sun

干。 dry. 批号:20080913。 Lot number: 20080913.

[0106] (2)拉萨大黄提取物制备:取拉萨大黄干燥根茎粉末100g,采用95%乙醇作为提取溶剂回流提取3次(2. 4L/1. 8L/1. 2L) , 120min/次,合并提取液,浓縮,真空干燥,得拉萨大黄提取物。 [0106] (2) Preparation of la rhubarb extract: Take dried rhizome of rhubarb powder Lasa 100g, using 95% ethanol as the extraction solvent reflux extraction three times (2. 4L / 1 8L / 1 2L..), 120min / times, combined The extract was concentrated, and dried in vacuo to give la rhubarb extract.

[0107] (3)纯化:拉萨大黄提取物2g,以40g 200目硅胶拌,采用硅胶柱层析纯化。 [0107] (3) Purification: la rhubarb extract 2g, 40g 200 mesh silica to mix, purified by chromatography on a silica gel column. 采用 use

氯仿/甲醇(V/V,8/1)作为洗脱溶剂,分部收集不同流份,薄层层析(TLC)监测,合并含有 Chloroform / methanol (V / V, 8/1) as the eluting solvent, collecting different fractions segment, thin layer chromatography (TLC) monitoring, fractions containing

曲札芪苷的流分,减压回收溶剂,15%乙醇重结晶,真空干燥,得曲札芪苷纯品。 Qu sheaf stilbene glycosides fractions under reduced pressure to recover the solvent, was recrystallized from 15% ethanol, and dried in vacuo to give pure music sheaf stilbene glycosides.

[0108] 实施例2中所述曲札芪苷提取物2g,按16 : 1加入纯水(32ml) ,6(TC加热溶解, [0108] Example 2 in the curved sheaf stilbene glucoside extract 2g, 16: 1 was added water (32ml), 6 (TC dissolved by heating,

放置4小时析晶,抽滤,晶体加入适量水,O. 5%活性碳脱色,重结晶,得1. 25g曲札芪苷,真 4 hours crystallization, filtration, water was added q.s. crystals, O. 5% activated bleaching, recrystallization, to give 1. 25g song sheaf stilbene glycosides, true

空干燥,得曲札芪苷纯品。 Air dried to give pure music sheaf stilbene glycosides.

[0109] (4)HPLC检测:按实施例1中HPLC法检测所得曲札芪苷的纯度,其不同流份产物中去氧土大黄苷的纯度可在90%〜99.9%之间;结晶-脱色-重结晶所得曲札芪苷纯度在98%〜99. 6%之间。 [0109] (4) HPLC Detection: Example 1. HPLC assay the purity of the resulting curve sheaf stilbene glycosides, different parts of which the purity of the product stream deoxy rhaponticin may be between 90% ~99.9%; crystallization - bleaching - resulting recrystallized song sheaf stilbene glycosides purity between 98% ~99 6%. HPLC检测典型图谱见图7。 Typical HPLC detection map shown in Figure 7. [0110] (5)曲札芪苷结构确认 [0110] (5) stilbene glycosides Structural confirmation curved sheaf

[0111] 检测所得曲札芪苷的UV、 negative FABMS、MS?H NMR、13CNMR谱图,其检测数据如下: ? [0111] detecting the resulting curve sheaf stilbene glycosides UV, negative FABMS, MS H NMR, 13CNMR spectrum, which detects data are as follows:

[0112] 曲札芪苷:白色毛状晶体(水),mp. 220-225 °C。 [0112] Qu sheaf stilbene glycosides: hairy white crystals (water), mp 220-225 ° C. 紫外灯下呈蓝紫色荧光。 Bluish violet color fluorescence under UV light. UV入max(Me涯):217 (4. 38) , 320 (4. 48) ;EI_MS(m/z) :406 [M]+, 244 [M_glycosy] + ;negative FAB-MS (m/z) :405 [M_H]—, 243 [MH-glycosy]—; 1H画R(CD30D, 500M Hz) S ppm :7. 44(1H,d, J = 2. 0Hz,C2' _H) , 7. 05(1H,dd, ^ = 2. OHz, J2 = 8. 0Hz,C6' _H) ,6. 92(lH,d,J = 16. 3Hz,CH) ,6. 83(1H,d, J = 16. 3Hz,CH) ,6. 82(1H,d, J = 8. 3Hz ,C5' _H) ,6. 47(1H,d, J = 2. 2Hz, C2,6-H) ,6. 18(1H, t, J = 2. 2Hz, C4_H) ,4. 81(1H, d, J = 7. 4Hz, anomericH),3.95(1H, dd, 1 = 2.0, 2. lHz, J2 = 12. 2Hz, C6 〃 —H),3.74(1H, dd, ^ = 6. OHz, J2 ==12. lHz, C6〃 -H) ,3. 41-3. 56(4H, m sugar-H) ;13C NMR(CD30D, 125MHz) S ppm :141. 1 (C_l),105. 9(C-2,6) , 159. 4(C-3,5) , 102. 7(C-4) , 131. 3(C-1' ),116.6(C_2' ),146.9(C_3'),148.1(C-4' ),117.2(C-5' ),123.0(06' ) , 127. 8 (C_ a ) , 129. 1 (C_ P ) , 104. 4 (C_l 〃 ),74.9(C-2〃 ),78.4(C-3〃 ),71.4(C_4" ),77.5(C_5〃 ) The UV max (Me ya): 217 (4. 38), 320 (4. 48); EI_MS (m / z): 406 [M] +, 244 [M_glycosy] +; negative FAB-MS (m / z) : 405 [m_H] -, 243 [MH-glycosy] -; 1H Videos R (CD30D, 500M Hz) S ppm:. 7 44 (1H, d, J = 2. 0Hz, C2 '_H), 7. 05 ( 1H, dd, ^ = 2. OHz, J2 = 8. 0Hz, C6 '_H), 6. 92 (lH, d, J = 16. 3Hz, CH), 6. 83 (1H, d, J = 16. 3Hz, CH), 6. 82 (1H, d, J = 8. 3Hz, C5 '_H), 6. 47 (1H, d, J = 2. 2Hz, C2,6-H), 6. 18 (1H , t, J = 2. 2Hz, C4_H), 4. 81 (1H, d, J = 7. 4Hz, anomericH), 3.95 (1H, dd, 1 = 2.0, 2. lHz, J2 = 12. 2Hz, C6 ... 〃 -H), 3.74 (1H, dd, ^ = 6. OHz, J2 == 12 lHz, C6〃 -H), 3 41-3 56 (4H, m sugar-H); 13C NMR (CD30D , 125MHz) S ppm:.. 141 1 (c_l), 105 9 (C-2,6), 159. 4 (C-3,5), 102. 7 (C-4), 131. 3 (C- 1 '), 116.6 (C_2'), 146.9 (C_3 '), 148.1 (C-4'), 117.2 (C-5 '), 123.0 (06'), 127. 8 (C_ a), 129. 1 ( C_ P), 104. 4 (c_l 〃), 74.9 (C-2 〃), 78.4 (C-3〃), 71.4 (C_4 "), 77.5 (C_5〃) 62.5(C_6〃 )。[0113] 通过与文献的比较,可以看出,本检测结果中各质子的化学位移数值与偶合常数的数值均与文献中(Y. Kashiwada, et al. I solation andCharacterization ofStilbenes. Chem. Pharm. Bull. 1984,32(9) :3501-3517 ;郑俊华,果德安。 62.5 (C_6〃). [0113] by comparison with the literature, it can be seen, the chemical shift value and the present value of the coupling constant in each of the detection results in the literature are protons (Y. Kashiwada, et al. I solation andCharacterization ofStilbenes . Chem Pharm Bull 1984,32 (9):... 3501-3517; Zheng Junhua, Dean fruit. 大黄现代研究.北京大学医学出版社,2007,第一版,286)的相应数值吻合。 Rhubarb modern research. Peking University Medical Press, 2007, first edition, 286) of the corresponding values ​​match. 曲札芪苷鉴定为(E)-l-(3,5-二羟苯基)-2- (3-羟基-4-0- -D-吡喃葡萄糖苯基)乙烯 Qu sheaf stilbene glycosides was identified as (E) -l- (3,5- dihydroxyphenyl) -2- (3-hydroxy-phenyl -4-0- -D- glucopyranose) ethylene

[0114] [(E)_1_ (3, 5_dihydroxyphenyl)_2_ (3_hydroxyl_4_0_ P _D_glucopyranyphenyl)ethylene],或称为3,5,3' ,4'-四羟基芪_3' _0_ P -葡萄糖苷(3, 5, 3' ,4' -tetra hydroxystilbene-3' -OP _D_glucopytanoside)。 [0114] [(E) _1_ (3, 5_dihydroxyphenyl) _2_ (3_hydroxyl_4_0_ P _D_glucopyranyphenyl) ethylene], otherwise known as 3,5,3 ', 4'-tetrahydroxy-stilbene-_3' _0_ P - glucoside (3, 5 , 3 ', 4' -tetra hydroxystilbene-3 '-OP _D_glucopytanoside). [0115] 实施例4 :拉萨大黄提取物的药理学作用[cms] (1)试验样品 [0115] Example 4: Pharmacological effect of la rhubarb extract [cms] (1) Test sample

[0117] 曲札芪苷提取物:实验代号QZQ-OO-Ol。 [0117] Qu sheaf stilbene glucoside extract: experiment code QZQ-OO-Ol. 样品按照实施例2所述步骤制备。 Samples were prepared as described in Example 2 step. 其中曲札芪苷含量为78.5% Wherein Sapporo stilbene glycosides song content 78.5%

[0118] 曲札芪苷:实验代号QZQ。 [01] stilbene glycosides song Sapporo: Experiment code QZQ. 样品按照实施例3所述步骤有曲札芪苷提取物制备,纯度为99. 1%。 Samples according to Example 3 has a curved step sheaf stilbene glycosides Extract preparation, purity of 99.1%.

[0119] 虎杖苷:购自西安华萃生物技术有限公司,批号:P0L050817,纯度为98.0X。 [0119] Polydatin: purchased from Cui Xian Biotechnology Co., Ltd., batch number: P0L050817, purity of 98.0X. [0120] 上述试验样品均以-羟丙基环糊精配制成不同浓度的供试溶液。 [0120] The test samples are - hydroxypropyl cyclodextrins formulated test solutions of different concentrations. [0121] (2)试验动物 [0121] (2) Test Animals

[0122] SPF级ICR小鼠,购自于昆明医学院实验动物中心。 [0122] SPF grade ICR mice were purchased from Kunming Medical Experimental Animal Center. [0123] SPF级SD大鼠,购自于昆明医学院实验动物中心。 [0123] SPF SD rats, purchased from Kunming Medical Experimental Animal Center. [0124] (3)试验方法[0125] 对大鼠MCAO局灶性脑缺血的影响 [0124] (3) Test Method [0125] Effect of focal cerebral ischemia in the rat MCAO

[0126] 各组雄性SD大鼠动物每日按剂量灌胃给药一次,连续5天。 [0126] Male SD rats of each group of animals at a dose gavage once daily for 5 consecutive days. 末次灌胃后30min, 用12%水合氯醛腹腔注射350mg/kg麻醉动物,左侧卧位,沿右耳眼线中点切开皮肤,分离颞肌,绞断颧骨,在颧骨根前方用牙科钻钻孔,暴露MCAO,在大脑下静脉和嗅束间用针挑起, 断之,棉球压迫止血后,分层缝合肌肉和皮肤。 After the last gavage 30min, with 12% chloral hydrate injected intraperitoneally 350mg / kg the animals were anesthetized, the left lateral position, right ear skin incision along the midpoint of the liner, the separation temporalis muscle, snare cheekbones, in front cheekbones Root dental drill then exposure of MCAO, provoke intravenous needle and olfactory tract at the brain, off, the cotton after hemostasis, layered suture muscle and skin. 假手术组除了不挑断MCAO外,其余步骤相同。 Except that Tiaoduan MCAO, the remaining steps same as sham operation group. 动物于造模后6h及24h根据运动等行为表现进行神经功能评分,以此作为脑功能障碍指标。 Animals at 6h and 24h after modeling exercise performance based on the conduct neurological score, as indicators of brain dysfunction. 评分采用盲法,即评分者不知道给药的情况, 0127] 评分标准如下: Rating blinded, i.e., does not know the situation score of administration, 0127] Rating criteria were as follows:

0128] 症状行为障碍评分 0128] symptoms of conduct disorder scores

0129] _ 0129] _

0130] 0131] 0132] 0133] 0134] 0135] 0136] 0137] 0138] 0130] 0131] 0132] 0133] 0134] 0135] 0136] 0137] 0138]

提鼠尾离开地面约一尺,手术对侧前肢出现腕屈曲,肘屈曲,肩内旋1〜4 或有腕,肘屈曲又有内旋。 Mention rat tail about one foot off the ground, forelimb contralateral surgical carpal flexion, elbow flexion, wrist or spin ~ 4, there are elbow flexion shoulder internal rotation.

将动物置于平地面上,分别推双肩向内侧,检查阻力。 Animals were placed in flat face, shoulders pushed inward, respectively check resistance. 1〜3 将动物置于金属网面上,观察两前肢的张力。 The animals were placed ~ 3 metal surface, the two front legs of tension was observed. 广3 将动物置于平地面上,观察有无转圈。 The animals were placed 3 wide flat surface to observe whether a circular motion. 广2 满分12 Canton 2 out of 12

以上标准满分为12分,分数越高,表明动物行为障碍越严重 The above criteria out of 12 points, with higher scores indicating more severe animal behavior disorders

造模24h,经神经功能评分后,颈动脉插管取血, 一份分离血清按试剂盒方法测定AST及LDH的活性;一份用3. 8%枸橼酸钠以1 : 9抗凝,1000rpm离心5min制备富血小板血浆(PRP) ,3500rpm离心10min制备贫血小板血浆(PPP)。 Modeling 24h, after neurological score, carotid artery blood, a measurement of AST and LDH activity according to the kit method of separating serum; with a 3.8% sodium citrate to 1: 9 anticoagulant, 5min centrifugation 1000rpm prepared platelet rich plasma (PRP), 3500rpm centrifuge 10min prepared platelet poor plasma (PPP). 按比浊法测定ADP-2Na诱导的血小板聚集率,ADP的终浓度为6mol/L ;按试剂盒方法分别测定TT、 PT、 APTT及FIB。 Induced by ADP-2Na Turbidimetry platelet aggregation, ADP at a final concentration of 6mol / L; measured according to kit Methods TT, PT, APTT, and FIB. 取血后快速断头取脑,放入冰生理盐水中去除嗅球、小脑和低位脑干,吸干水分称取脑重,沿冠状切成5片,置入5ml含有4% TTC及lmol/LK2HP040. lml的溶液中,避光于37"温孵30min,其间每隔7-8min翻动一次,经染色后,正常脑组织呈玫瑰红色,而梗死组织呈白色,将梗塞部位剪下称重,以重量求面积法计算梗死组织重量占双侧大脑半球重的百分比作为脑梗塞范围。 Fast decapitated brain after blood, placed in an ice saline removed olfactory bulb, cerebellum and lower brain stem, brain weight drain water was weighed, cut into 5 coronal direction, placed 5ml comprising 4% TTC and lmol / LK2HP040 . lml of a solution, protected from light at 37 "incubated 30min, during which every 7-8min turned over once dyed, the normal brain tissue showed rose color, while the infarct tissue showed white, the infarction area cut weighed to wt mensuration method to calculate infarct tissue weight accounts for both hemispheres infarct as a percentage of weight.

[0139] 对胶原蛋白-肾上腺素所致小鼠血栓形成影响取20〜22g小鼠,雌雄各半,按性别和体重随机分组,见表6。 [0139] collagen - induced thrombosis in mice Effects of epinephrine taken 20~22g mice, male and female, were randomized by sex, and weight, see Table 6. 各组动物每日按剂量腹腔注射给药一次,连续5天。 Each dose group were injected intraperitoneally daily press, for 5 consecutive days. 末次给药后30min,每鼠按0. lml/10g体重的剂量静脉注射由胶原蛋白-肾上腺素制成的诱导剂, 观察并记录15min内各组动物偏瘫发生率、死亡率及恢复率。 30min, each mouse press 0. lml / 10g body weight by intravenous dose of collagen after the last administration - inducing agent made of adrenaline Groups of animals were observed and recorded hemiplegia, mortality and recovery rate in 15min. X2检验组间差异的显著性。 Significant differences between the groups X2 test. [0140] 对冠脉结扎致大鼠心肌缺血影响 [0140] Ischemia Myocardial coronary ligation

[0141] 雄性SD大鼠,330〜385g,按体重随机分组见表7,各组动物按剂量每日灌胃给药一次,连续5天,空白组和模型组给予等容量0. 5% CMC-Na 10ml/kg体重。 [0141] Male SD rats, 330~385g, randomized by body weight are shown in Table 7, each group of animals at a dose gavage once daily for 5 consecutive days, group and untreated group were given the same volume 0. 5% CMC -Na 10ml / kg body weight. 末次给药后30min,用12%水合氯醛腹腔注射350mg/kg麻醉动物,行开胸手术暴露心脏,于肺动脉圆锥及左心房间找出冠状动脉左前降支,距根部2〜3mm处结扎,快速心脏复位关闭胸腔。 After the last administration 30min, injection with 12% chloral hydrate 350mg / kg the animals were anesthetized, thoracotomy to expose the heart, to the left atrium and the pulmonary cone find anterior descending coronary artery, from 2~3mm ligated at the root, fast closed chest cardiac reset. 假手术组仅置线头而不结扎,造成急性心肌缺血模型,24h后,颈动脉插管取血分离血清按试剂盒方法测定AST、LDH及CK-MB的活性;取血后立即取心脏,置于冰生理盐水中泵出心腔内积血,去掉心房及脂肪组织,吸干水分称取心室重,随后沿冠状切成5片,在1% TTC溶液中避光于37t:孵育染色10min。 Sham group only opposing thread without ligation model of acute myocardial ischemia, 24h later, the carotid artery was measured take AST, LDH and CK-MB activity according to the kit method of separating blood serum; blood taken immediately after the heart, placed in an ice saline pumped intracardiac blood clots, and fatty tissue removed atrial, ventricular weight drain water was weighed and then cut into 5 coronal direction, in 1% TTC solution was protected from light to 37t: 10min incubation staining . 经染色后,正常心肌组织呈红色,而梗死组织呈白色,将梗塞部位剪下称重,计算坏死部分占心室重的百分比为心肌梗塞范围。 After staining, normal myocardial tissue red, white and infarcted tissue, the infarction area cut weighed to calculate the weight of necrotic portion comprises a percentage of ventricular myocardial infarction range. [0"2] (5)试验结果 [0 "2] (5) Test results

[0143] 对大鼠中动脉结扎(MCA0)致局灶性脑缺血的影响 Effect of focal cerebral ischemia [0143] induced by cerebral artery ligation (MCA0)

[0144] 曲札芪苷提取物以50, 100mg/kg,曲札芪苷纯品以15, 30mg/kg的剂量分别于MCA0 手术后即刻和缺血再灌注6h静脉注射2次,能明显减轻大鼠局灶性脑缺血所致的神经功能障碍,明显减小脑梗塞的范围,曲札芪苷提取物两剂量组分别縮小65. 87% 、40. 38% ;曲札芪苷纯品两剂量组分别縮小75. 87%、52. 21%,优于虎杖苷。 [0144] Qu sheaf stilbene glucoside extract is 50, 100mg / kg, pure music sheaf stilbene glycosides to 15, 30mg / kg doses, respectively, immediately after surgery and MCA0 ischemia reperfusion 6h intravenously twice, can significantly reduce neurological disorders caused by focal cerebral ischemia, cerebral infarction significantly reduced range, Qu Sapporo stilbene glucoside extracts two dose reduction 65.87%, 40 38%, respectively; Qu Sapporo pure stilbene glucoside two dose groups were 75.87% reduction, 52.21%, better polydatin.

[0145] 表4曲札芪苷纯品及提取物对MCA0再灌注大鼠神经功能障碍及脑梗塞范围的影 [0145] Table 4 sheaf stilbene glucoside extract and pure MCA0 Movies reperfusion disorders and neurological function of cerebral infarct

响(X士SD) Sound (X disabilities SD)

[0146] [0146]

14<table>table see original document page 15</column></row> <table> 14 <table> table see original document page 15 </ column> </ row> <table>

[0147] 与假手术组相比:A AP < 0. 01 ;与模型组相比:*P < 0. 05, **P < 0. 01 ; [0147] Compared with the sham group: A AP <0. 01; compared with model group: * P <0. 05, ** P <0. 01;

[OH8] 表5曲札芪苷纯品及提取物对MCAO再灌注大鼠血凝及血小板聚集的影B向(;±SD) [OH8] Table 5 sheaf stilbene glycosides and extracts pure MCAO reperfusion rats on blood coagulation and platelet aggregation Movies to B (; ± SD)

[0149] [0149]

<table>table see original document page 15</column></row> <table>[0150] 与假手术组相比:AP < 0. 05 ;与模型组相比:*P < 0. 05, **P < 0. 01 ; <Table> table see original document page 15 </ column> </ row> <table> [0150] Compared with sham operation group: AP <0. 05; compared with model group: * P <0. 05, * * P <0. 01;

[0151] 对胶原蛋白_肾上腺素所致小鼠血栓形成影响 [0151] _ epinephrine collagen induced thrombosis in mice Effect

[0152] 曲札芪苷提取物60mg/kg、120mg/kg剂量组,曲札芪苷纯品各剂量组均能明显抑 [0152] Qu sheaf stilbene glucoside extract 60mg / kg, 120mg / kg dose group, stilbene glycosides pure music sheaf each dose could significantly suppressed

制胶原-肾上腺素所致的小鼠体内血栓形成;。 Manufactured by Collagen - epinephrine-induced thrombosis in mice in vivo;.

[0153] 表6曲札芪苷纯品及提取物对小鼠体内血栓形成的影响 [0153] Table 6 Effect sheaf pure stilbene glucoside extract and thrombus formation in mice

[0154]<table>table see original document page 16</column></row> <table>[0155] 与对照组相比:*P < 0. 05, **P < 0. 01 [0156] 对冠脉结扎致大鼠急性心肌梗塞范围的影响[0157] 模型组大鼠血清AST、 LDH及CK-MB活性有不同程度的增高,尤以CK-MB及LDH活性增高更明显,表明心肌损伤造模成功。 [0154] <table> table see original document page 16 </ column> </ row> <table> [0155] Compared with the control group: * P <0. 05, ** P <0. 01 [0156] of Effects of acute myocardial infarct size coronary ligated rats [0157] rats in model group of AST, LDH and CK-MB activity has increased in different degrees, particularly CK-MB and LDH activity increased more significantly, indicating myocardial injury made successful model. 曲札芪苷(50mg/kg)及曲札芪苷提取物(200mg/ kg)均能明显降低CK-MB和LDH的释放,表明减轻心肌细胞损伤作用显著;两给药组大鼠的心肌梗塞范围均显著小于模型对照组,与模型组相比分别縮小64. 33%和47. 72% 。 Qu Sapporo stilbene glucoside (50mg / kg) and curved sheaf stilbene glucoside extract (200mg / kg) could significantly reduce the release of LDH and CK-MB, reduce myocardial cell injury showed a significant effect; MI rats administered two They were significantly less than the range of the model control group, 64.33% reduction and 47.72%, respectively, compared with model group. 结果见表7。 The results are shown in Table 7. [0158] 表7. QHQ及QZQ对大鼠心肌梗塞范围和酶学的影响[0159]<table>table see original document page 16</column></row> <table>[0160] 实施例5几种芪苷类物质的DPPH自由基清除活性试验[0161] (1)试验样品 [0158] Table 7. Effect on Myocardial Infarction QHQ and QZQ range and enzymology [0159] <table> table see original document page 16 </ column> </ row> <table> [0160] Example 5 several species stilbene glycosides substances DPPH radical scavenging activity test [0161] (1) test sample

[0162] 曲札芪苷:自制,样品按照实施例3所述步骤有曲札芪苷提取物制备, [0163] 纯度为99. 1%。 [0162] Qu sheaf stilbene glycosides: self, the sample according to Example 3 was prepared with a curved step sheaf stilbene glycosides extracted, [0163] purity of 99.1%.

[0164] 虎杖苷:购自西安华萃生物技术有限公司,批号:P0L050817,纯度为98.0X。 [0164] Polydatin: purchased from Cui Xian Biotechnology Co., Ltd., batch number: P0L050817, purity of 98.0X. [0165] 土大黄苷:自制,按中国发明专利200510010757.X中所述土大黄苷纯品制备,纯度:99. 5%。 [0165] rhaponticin: self, according to the Chinese invention patent 200510010757.X rhaponticin in pure preparation, purity: 995%.

[0166] 大黄降脂素:自制,从河套大黄(Rheum hotaoense CYCheng et CT Kao)中制备,纯度:99.0% [0166] Rhapontinum: self-made, prepared from Loop rhubarb (Rheum hotaoense CYCheng et CT Kao), Purity: 99.0%

[0167] 维生素E(-Tocpherol) 、 BHA(butylated hydroxyanisol) 、 DPPH自由基 [0167] Vitamin E (-Tocpherol), BHA (butylated hydroxyanisol), DPPH radical

(,-diphenyl- -picrylhydrazyl)均为Sigma产品。 (, -diphenyl- -picrylhydrazyl) were purchased from Sigma products. 其中,维生素E为常用的天然来源的 Among them, vitamin E is a common source of natural

抗氧化剂,BHA为食品工业中常用的合成抗氧化剂。 Antioxidants, BHA for the food industry commonly used synthetic antioxidants.

[0168] 上述试验样品均以甲醇配制成不同样品浓度的供试溶液。 [0168] The test samples are prepared in methanol solutions of different test sample concentration.

[0169] (2)试验方法 [0169] (2) Test Method

[0170] 自由基清除活性测定参照MH Gorden等人的方法(MH Gordon, et al. Antioxidant activity of hydroxytyrosol acetate compared with that of otheroliver oil polyphenols. J. Agric. Food Chem. , 2001, 49 :2480)。 [0170] The method of free radical activity was measured referring MH Gorden et al clearance (MH Gordon, et al Antioxidant activity of hydroxytyrosol acetate compared with that of otheroliver oil polyphenols J. Agric Food Chem, 2001, 49:.... 2480). 每一测定的试液为0. 06Mm DPPH自由基甲醇溶液.参照品及供试样品均配成2. 1 (0. 06X35)mM的甲醇溶液,分别稀释成1/10, 1/4, 1/2三个不同的浓度。 Each test solution was determined to be 0. 06Mm DPPH radical solution in methanol. Reference standard and test samples were formulated as methanol 2. 1 (0. 06X35) mM respectively diluted to 1/10, 1/4, 1 three different concentrations / 2. 测定时,在3.5ml DPPH自由基甲醇溶液中加入O. 1ml参照品或样品溶液,混合,在15分钟测定其光吸收值(A,pJ,检测波长为515nm,每个不同浓度的参照品及样品平行测定三次。甲醇为空白,0.06mM DPm溶液为对照(Control)。每一参照品和供试样品得到3-4个不同浓度分别在不同时间的三组平行 Measurement, O. 1ml added to the sample or reference solution, mixed solution of methanol and 3.5ml DPPH radical, which measured absorbance (A, pJ, detection wavelength of 515nm within 15 minutes of each reference standard and various concentrations of three samples were assayed in parallel. methanol as blank, 0.06mM DPm solution as a control (control). each test sample and the reference were obtained 3-4 different concentrations of three sets of parallel, respectively, at different times

测定值。 measured value. 每一测定值表示DPra自由基在溶液中的残留值,用以下公式将其换算成DPra Each value represents the measured value DPra residual free radicals in solution, with the following formula, which is converted into DPra

自由基清除率(scavenged DPPH radical rate% ) :Scavenged DPPH radical rate% = Radical scavenging (scavenged DPPH radical rate%): Scavenged DPPH radical rate% =

control nsample^ / ncontro]y control nsample ^ / ncontro] y

[oi7i] 以此值作出样品在不同浓度时DPra自由基清除率与时间的关系图及在不同时间 [Oi7i] In this graph the sample value to DPra radical scavenging time and at different concentrations and at different times

下DPra自由基清除率与样品浓度的关系。 DPra relation radical scavenging concentration in the sample. 样品浓度用加入样品与DPPH自由基的摩尔数比值(molar ratio)来表示,本实验中取O. 10, 0. 25, 0. 50,1.00。 The molar ratio of the number of sample added to the sample and the concentrations of DPPH radical (molar ratio) is represented, in this experiment taken O. 10, 0. 25, 0. 50,1.00. 样品清除自由基的活性EC5。 EC5 radical scavenging activity of the sample. 是指使DPra自由基被清除50%所需的样品摩尔数与加入DPra摩尔数的比值。 Refers to a radical DPra cleared moles of 50% and the ratio of the sample required DPra added moles. [0172] (3)试验结果 [0172] (3) Test Results

[0173] 表8cophero1、 BHA及各芪苷类化合物DPffl自由基活性[0174] [0173] Table 8cophero1, BHA and various stilbene glycosides DPffl radical activity [0174]

<table>table see original document page 17</column></row> <table>[0175] 表8数据显示,曲札芪苷清除DPra自由基的活性较强。 <Table> table see original document page 17 </ column> </ row> <table> [0175] Data in Table 8 show, curved sheaf stilbene glycosides clear DPra radical strong activity. [0176] 实施例6曲札芪苷片剂制备 Preparation of tablets 6 cases sheaf stilbene glycosides [0176] Embodiment

[0177] 曲札芪苷提取物:样品按照实施例2所述步骤制备,其中提取溶剂采用90%乙醇;其中曲札芪苷的含量为78.5%。 [0177] Qu sheaf stilbene glucoside extract: the samples prepared according to Example 2 step, wherein the 90% ethanol extraction solvent employed; wherein the content of the song sheaf stilbene glycosides was 78.5%.

[0178] 制法:称取曲札提取物100g,溶解于适量乙醇中,加入900g PVP K29/32,混合均匀,使充分溶解;旋转蒸发使乙醇挥发,冷却,干燥,固化,粉碎固化物,取100克与50克乳糖、30克预胶化淀粉、10克羧甲基淀粉钠混合,过80目筛2遍,混合均匀,以1 %低取代羟丙基纤维素乙醇溶液(浓度50%)为粘合剂制软材,过40目筛制粒,6(TC干燥,过30目筛整粒,加入1.5克滑石粉混合均匀,压片,可制备曲札芪苷片1000片。 [0179] 实施例7 :曲札芪苷冻干粉针 [0178] Method: Weigh song Zhati extract 100g, was dissolved in an appropriate amount of ethanol, was added 900g PVP K29 / 32, mixed evenly, so that fully dissolve; rotary evaporator to evaporate the ethanol, cooled, dried, solidified, pulverized cured, take 100 g of lactose, 50 g, 30 g pregelatinized starch, 10 g sodium carboxymethyl starch are blended, passed 2 times 80 mesh sieve, mixed with 1% low-substituted hydroxypropylcellulose in ethanol (concentration 50% ) made of soft material as a binder, granulated through a 40 mesh screen, 6 (TC dried, sieved through a 30 mesh sieve, mixed 1.5 g of talc, tabletting, may be prepared music piece 1000 sheaf stilbene glycosides. [ 0179] Example 7: lyophilized Powder song sheaf stilbene glycosides

[0180] 曲札芪苷纯品:样品按照实施例3所述步骤制备,其中提取溶剂采用90%乙醇;其中曲札芪苷的含量为99.0%。 [0180] Qu sheaf pure stilbene glycosides: the sample as in Example 3, Step prepared, wherein the extraction solvent using 90% ethanol; wherein the content of the song sheaf stilbene glycosides of 99.0%.

[0181] 配方: [0181] formula:

[0182] 曲札芪苷纯品10g [0182] Qu sheaf stilbene glucoside pure product 10g

[0183] 大豆磷脂8g [0183] soya lecithin 8g

[0184] 甘露醇200g [0184] Mannitol 200g

[0185] 制备工艺:在1万级的条件下,将合格的药物和辅料一起溶于注射用水中,经脱色 [0185] Preparation process: Under the conditions of 10,000, was dissolved in conformity with the pharmaceutical excipients and water for injection, decolored

等处理后,加入注射用水稀释至所2000ml 。 Once treated, diluted with water for injection is added 2000ml. 过滤,滤液经过滤纸及0. 4 ii m的滤膜粗滤等处 Filtered and the filtrate filter through the membrane and 0. 4 ii m straining, etc.

理后,在100级的条件下,用0. 2ym以下的滤膜过滤。 After treatment under the conditions of 100, with the following membrane filtration 0. 2ym. 精滤液送入分装机,按每支2ml分 The filtrate was installed into fine points, divided by each 2ml

装,盖上带槽的盖子,送入冷冻真空干燥机,快速冷至35°C〜40°C,2〜3小时,逐步缓慢升温 Installed, cover and grooved, frozen into a vacuum dryer, flash cooled to 35 ° C~40 ° C, 2~3 hours, slowly warmed gradually

至35t:〜45t:(约需10小时)。 To 35t: ~45t :( takes about 10 hours). 塞紧盖子,取出制品,扎盖、包装、检验、合格的药用粉针成 Stoppered cover and remove products, bar cap, packaging, testing, pharmaceutically acceptable powder injection into

品(1000支10mg之规格的冻干粉针)。 Goods (1000 specifications 10mg of freeze-dried powder).

[0186] 实施例8 :曲札芪苷口服液产品 [0186] Example 8: Liquid Product curved sheaf stilbene glycosides

[0187] 配方:脂溶性芪苷(含量98 % ) 48g [0187] Formulation: fat-soluble stilbene glycosides (content 98%) 48g

[0188] 蜂蜜1000g [0188] 1000g honey

[0189] 丙二醇500g [0189] 500g of propylene glycol

[0190] 蒸馏水加至10kg [0190] Distilled water was added to 10kg

[0191] 制备方法:取处方量药物和辅料,按普通口服液制备方法制备,可得IOOO支的口服液。 Preparation [0191] Method: The formulation amount of drug and excipients was prepared by the general method of oral liquid preparation, oral IOOO branch can be obtained.

Claims (9)

  1. 曲札茋苷在制备预防和治疗缺血性心脑血管疾病制剂中的应用,所述曲札茋苷为化合物(E)-1-(3,5-二羟苯基)-2-(3-羟基-4-O-β-D-吡喃葡萄糖苯基)乙烯,化学结构是:使用的曲札茋苷为曲札茋苷提取物。 Application of song Sapporo stilbene glycosides formulation for the prevention and treatment of ischemic cardiovascular disease, said curved Sapporo stilbene glucoside compound (E) -1- (3,5- dihydroxyphenyl) -2- (3 - hydroxy -4-O-β-D- glucopyranose phenyl) ethylene, the chemical structure: use of a stilbene glycosides Sapporo song for the song sheaf stilbene glucoside extract. FSA00000046829000011.tif FSA00000046829000011.tif
  2. 2. 权利要求1所述曲札芪苷在制备预防和治疗缺血性心脑血管疾病制剂中的应用其特征在于所述曲札芪苷提取物,按重量百分比计,曲札芪苷提取物中含有曲札芪苷的有效成份为50% -100%。 The sheaf a stilbene glycosides according to claim 1 characterized in that the application and therapeutic agent for the prevention of ischemic cardiovascular disease in that said curved sheaf stilbene glucoside extract, by weight percent, stilbene glycoside extract song Sapporo containing active ingredient song stilbene glycosides sheaf of 50% to 100%.
  3. 3. 权利要求1所述曲札芪苷提取物的一种制备方法,其特征在于所述制备方法包括下列步骤:(1) 前处理:前处理步骤包括拉萨大黄原植物根或根茎的采集、净制和/或干燥、切碎或粉碎;(2) 提取:所述提取是采用水、含水或不含水有机溶剂作为提取溶剂对步骤(1)所述拉萨大黄根或根茎进行提取获得拉萨大黄提取液;所述有机溶剂包括含有广4个碳原子的醇、酮、醚以及它们的混合物。 A process for preparing a sheaf of the stilbene glucoside extracts of claim 1, wherein said preparation method comprises the following steps: (1) Pre-treatment: pre-processing step includes acquiring the original la rhubarb roots or rhizomes of the plant, net system and / or dried, pulverized or chopped; (2) extracting: the extraction is the use of water, aqueous or non-aqueous organic solvent as an extraction solvent in step (1) the roots or rhizomes Rheum Lhasaense la rhubarb extract obtained extract; wide containing the organic solvent comprises an alcohol four carbon atoms, ketones, ethers, and mixtures thereof. (3) 后处理:后处理步骤包括回收或部分回收步骤(2)所得提取液中的有机溶剂得到提取浓縮液,所得提取浓縮液可加入或不加水,继而将提取液进行静置沉淀,固液分离除去部分杂质;除杂质后的液相经进一步吸附树脂层析纯化、分段洗脱、洗脱物浓縮析晶获得到以重量百分比计,含有曲札芪苷为50% _100%的曲札芪苷提取物。 (3) Processing: or post-treatment step comprising recovering part of the recovery step (2) the resulting organic extract was concentrated to obtain an extraction solvent, the resulting extract concentrate may be added or without addition of water, then extract liquid is left to settle , solid-liquid separation to remove part of the impurities; other liquid impurities after further purified by chromatography on adsorption resin, segment elution was concentrated to obtain crystallization in weight percent, comprising curved stilbene glycosides sheaf 50% _100 Qu sheaf% stilbene glucoside extract.
  4. 4. 根据权利要求3所述曲札芪苷提取物的一种制备方法,其步骤(2)所述有机溶剂为甲醇、乙醇、丙酮、正丁醇或乙醚;其步骤(2)所述提取溶剂为50〜100%的乙醇或甲醇水溶液。 4. A process for preparing the 3 sheaf stilbene glycoside extract Claim step (2) said organic solvent is methanol, ethanol, acetone, n-butanol or ether; which step (2) the extraction 50~100% of the solvent is ethanol or aqueous methanol.
  5. 5. 曲札芪苷纯品的一种制备方法,其特征在于,所述制备方法是采用拉萨大黄作为原料,通过提取纯化步骤获得所需曲札芪苷纯品;所述曲札芪苷纯品纯度以重量百分比计,为90% -100%。 5. A process for preparing stilbene glycosides song sheaf pure product, wherein the preparation method is employed as a starting material la rhubarb, Sapporo stilbene glucoside obtained pure music desired by extraction and purification steps; the song stilbene glycosides pure Sapporo purity products in weight percentage, of 90% -100%.
  6. 6. 根据权利要求3所述曲札芪苷提取物的一种制备方法,其特征在于曲札芪苷提取物是从野生或栽培的拉萨大黄(Rheum lhasaense AJ Li et PK Haiao)的根或根茎中提取。 A method of preparing the sheaf 3 stilbene glycoside extract according to claim, characterized in that the curved sheaf stilbene glycoside extract from wild or cultivated la rhubarb (Rheum lhasaense AJ Li et PK Haiao) roots or rhizomes extract from.
  7. 7. 曲札芪苷在制备曲札芪苷纯品中的用途。 7. Qu Qu Sapporo stilbene glycosides prepared sheaf stilbene glycosides use of pure.
  8. 8. 曲札芪苷提取物在制备预防或治疗心脑血管疾病制剂中的用途。 8. Qu sheaf stilbene glycoside extract for the prevention or treatment of cardiovascular and cerebrovascular diseases formulation.
  9. 9. 曲札芪苷制剂作为各种医药上可接受的制剂或保健食品,可用于制作药物或保健食 9. Qu sheaf stilbene glycosides of various formulations as a pharmaceutically acceptable formulation or health food, can be used to produce drugs or health food
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