CN103906525B - 楹树提取物在制备治疗胃溃疡药物中的应用 - Google Patents
楹树提取物在制备治疗胃溃疡药物中的应用 Download PDFInfo
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Abstract
一种楹树提取物及其制备方法，所述楹树提取物用于制备质子泵抑制剂和治疗酸相关性疾病，如胃溃疡和慢性胃炎的药物。
Description
技术领域
本发明涉及一种楹树[Albizia chinensis(Osbeck)Merr.]中提取物及其制备方法，本发明还涉及楹树提取物作为质子泵抑制剂在制备治疗胃溃疡和酸相关性疾病药物中的医药用途，医药技术领域，。
背景技术
胃溃疡是消化***常见的疾病，且易复发。随着现代生活节奏加快，精神紧张和工作压力增大，使得胃溃疡的发病率呈上升趋势，危害及人们的身心健康。目前临床上应用较多的抗溃疡西药主要为雷尼替丁、奥美拉唑等药物，虽然西药近期疗效明显，但其存在服药后出现不良反应和毒副作用。因此，人们开始研发高效、低毒的新型的抗溃疡药物。基于我国传统中药的长期临床实践和用药历史，从传统中草药中寻找和挖掘具有抗胃溃疡的天然活性成分或有效组分，具有先天优势和广阔的临床应用价值。
楹树Albizia chinensis(Osbeck)Merr.为豆科合欢属(Albizia)植物，其药用部位为树皮，树皮含鞣质，有固涩止泻、收敛生肌之功效，主治肠炎、腹泻、痢疾(全国中草药汇编，全国中草药汇编编写组编，第二版，下册，1986年，PP768)。另有记载楹树皮含三萜皂苷(又称合欢催产素)，有收缩子宫的作用(中药大辞典，江苏新医学院编，1998年，上册，PP937)。近年来，我们从楹树皮中分离得到一系列结构复杂的具有肿瘤细胞毒活性的三萜皂苷(J.Nat.Prod.2009，72，632-639；Carbohydr.Res.2010，345，1877-1881)。此外，我们还发现楹树Albizia chinensis提取物具有镇静催眠的活性(中国专利公开号CN101766676A)。迄今为止尚未见有关楹树皮提取物在抑制质子泵活性和治疗胃溃疡或慢性胃炎方面的药理活性研究的报道。
发明内容
本发明要解决的技术问题是，提供一类新的治疗胃溃疡和酸相关性疾病的药物，即楹树Albizia chinensis(Osbeck)Merr.提取物。本发明的楹树Albizia chinensis(Osbeck)Merr.提取物可作为质子泵抑制剂，用于制备治疗胃溃疡和酸相关性疾病药物。
本发明要解决的另一个技术问题是提供制备楹树提取物的方法以及这种方法获得的楹树提取物。
本发明要解决的又一个技术问题是提供一种药物组合物，包括作为活性成分的楹树提取物及制药领域中常用的载体。
本发明要解决的再一个技术问题是提供楹树提取物Albizia chinensis(Osbeck)Merr.在制备治疗胃溃疡和酸相关性疾病药物中的应用。
解决本发明的技术问题，本发明采取如下的技术方案：
本发明涉及一种可用于制备质子泵抑制剂治疗胃溃疡和酸相关性疾病药物的楹树提取物，其制备方法为：
(1)将楹树原药材粉碎并用溶剂提取，提取液浓缩得楹树浸膏。
优选的楹树原药材是楹树茎皮。楹树原药材经干燥并适当的粉碎，以利增大与溶剂的接触面积提高效率。
原药材的提取溶剂可以使用醇类溶剂或水与醇类的混合溶剂。
优选的醇类包括甲醇、乙醇、异丙醇、丁醇等，最优选的是乙醇。
乙醇浓度可以是体积比50-100％；优选的浓度为体积比80～95％。
提取时溶剂使用量是溶剂体积与原药材重量的体积/重量比(单位：升/公斤)为3～10∶1，优选是3～5∶1。
提取可以在静态或动态下进行，优选在动态条件下，例如搅拌。为了提高提取的效率，可以使用超声波等。
提取的温度是从室温(例如20℃)到溶剂回流温度的范围内，优选的温度是溶剂回流的温度下。
提取可连续或间歇进行，间歇提取时可重复1～5次，优选2-4次。
提取时间是1-5小时，优选为2-3小时。
合并提取液，在常压或减压加热浓缩成膏状，优选在减压条件下浓缩。
优选楹树浸膏的制备方法如下：称量粉碎的楹树茎皮药材，以药材重量计每公斤用3-5升的80-95％乙醇回流2-4次，每次2-3小时；合并提取液，提取液经减压浓缩得到楹树(Albizia chinensis(Osbeck)Merr)浸膏。
(2)楹树浸膏进一步通过聚酰胺柱色谱，通过醇类溶剂梯度洗脱，精制纯化得到楹树提取物。
楹树浸膏以10-30％的乙醇溶解，优选20％的乙醇溶解。
聚酰胺吸附剂干粉的用量以体积(升)与药材提取浸膏的重量(公斤)的比例为20～40倍，优选25～30倍。
洗脱剂可以醇类与水的混合溶剂；优选的醇类溶剂为甲醇和乙醇，最优选醇类溶剂为乙醇。
洗脱溶剂的用量为聚酰胺干粉的体积3-10倍，优选洗脱剂用量为聚酰胺干粉的体积4-5倍。
依次用10-30％的乙醇、40-80％的乙醇进行梯度洗脱，40-80％的乙醇洗脱液减压回收得到楹树提取物。
优选的聚酰胺柱色谱纯化步骤包括：将楹树浸膏以10-30％的乙醇溶解后，加到30～90目的聚酰胺柱上[聚酰胺吸附剂干粉的用量以体积(升)与药材提取浸膏的重量(公斤)的比例为25-30]，依次用10-30％的乙醇、40-80％的乙醇进行梯度洗脱，洗脱溶剂的用量为3-5倍聚酰胺干粉的体积，浓缩40-80％的乙醇洗脱液得到楹树提取物。
本发明涉及了一种药物组合物，包括以本发明所述的方法提取的楹树提取物及药学上可接受的载体。
本发明还涉及含有作为活性成份的本发明提取物和常规药物赋形剂或辅剂的药物组合物。通常本发明药物组合物含有0.1～95％重量的本发明提取物。
本发明还提供一种药物组合物，它包括药物有效剂量的，作为活性成分的如本发明方法提取的楹树提取物和/或楹树及药学上可接受的载体。
本发明提取物的药物组合物可根据本领域公知的方法制备。用于此目的时，如果需要，可将本发明提取物与一种或多种固体或液体药物赋形剂和/或辅剂结合，制成可作为人药或兽药使用的适当的施用形式或剂量形式。
本发明提取物或含有它的药物组合物可以单位剂量形式给药，给药途径可为肠道或非肠道，如口服、静脉注射、肌肉注射、皮下注射、腹腔注射、鼻腔、口腔粘膜、眼、肺和呼吸道、皮肤、***、直肠等，优选口服给药。
给药剂型可以是液体剂型、固体剂型或半固体剂型。液体剂型可以是溶液剂(包括真溶液和胶体溶液)、乳剂(包括o/w型、w/o型和复乳)、混悬剂、注射剂(包括水针剂、粉针剂和输液)、滴眼剂、滴鼻剂、洗剂和搽剂等。固体剂型可以是片剂(包括普通片、肠溶片、含片、分散片、咀嚼片、泡腾片、口腔崩解片)、胶囊剂(包括硬胶囊、软胶囊、肠溶胶囊)、颗粒剂、散剂、微丸、滴丸、栓剂、膜剂、贴片、气(粉)雾剂、喷雾剂等；半固体剂型可以是软膏剂、凝胶剂、糊剂等。
本发明提取物可以制成普通制剂、也可以是缓释制剂、控释制剂、靶向制剂及各种微粒给药***。
为了将单位给药剂型制成片剂，可以广泛使用本领域公知的各种赋形剂，包括稀释剂、黏合剂、润湿剂、崩解剂、润滑剂、助流剂。稀释剂可以是淀粉、糊精、蔗糖、葡萄糖、乳糖、甘露醇、山梨醇、木糖醇、微晶纤维素、硫酸钙、磷酸氢钙、碳酸钙等；湿润剂可以是水、乙醇、异丙醇等；粘合剂可以是，淀粉浆、糊精、糖浆、蜂蜜、葡萄糖溶液、微晶纤维素、***胶浆、明胶浆、羧甲基纤维素钠、甲基纤维素、羟丙基甲基纤维素、乙基纤维素、丙烯酸树脂、卡波姆、聚乙烯毗咯烷酮、聚乙二丙醇等；崩解剂可以是干淀粉、微晶纤维素、低取代羟丙基纤维素、交联聚乙烯毗咯烷酮、交联羧甲基纤维素钠、羧甲基淀粉钠、碳酸氢钠与构椽酸、碳酸钙、聚氧乙烯山梨糖醇脂肪酸酯、十二烷基磺酸钠；润滑剂和助流剂可以是滑石粉、二氧化硅、硬脂酸盐、酒石酸、液体石蜡、聚乙二醇等。
还可以将片剂进一步制成包衣片，例如糖包衣片、薄膜包衣片、肠溶包衣片，或双层片和多层片。
为了将给药单元制成丸剂，可以广泛使用本领域公知的各种载体。关于载体的例子是，例如稀释剂与吸收剂，如葡萄糖、乳糖、淀粉、可可脂、氢化植物油、聚乙烯吡咯烷酮、月桂酸聚乙二醇甘油酯、高岭土、滑石粉等；粘合剂，如***胶、黄菩胶、明胶、乙醇、蜂蜜、液糖、米糊或面糊等；崩解剂，如琼脂粉、干燥淀粉、海藻酸盐、十二烷基磺酸钠、甲基纤维素、乙基纤维素等。
为了将给药单元制成栓剂，可以广泛使用本领域公知的各种载体。关于载体的例子是，例如聚乙二醇、卵磷脂、可可脂、高级醇、高级醇的酯、明胶、半合成甘油酯等。
为了将给药单元制成胶囊，将有效成分本发明提取物与上述的各种载体混合，并将由此得到的混合物置于硬的明胶胶囊或软胶囊中。也可将有效成分本发明提取物制成微囊剂，混悬于水性介质中形成混悬剂，亦可装入硬胶囊中或制成注射剂应用。
例如，将本发明提取物制成注射用制剂，如溶液剂、混悬剂溶液剂、乳剂、冻干粉针剂，这种制剂可以是含水或非水的，可含一种和/或多种药效学上可接受的载体、稀释剂、粘合剂、润滑剂、防腐剂、表面活性剂或分散剂。如稀释剂可选自水、乙醇、聚乙二醇、1，3-丙二醇、乙氧基化的异硬脂醇、多氧化的异硬脂醇、聚氧乙烯山梨醇脂肪酸酯等。另外，为了制备等渗注射液，可以向注射用制剂中添加适量的氯化钠、葡萄糖或甘油，此外，还可以添加常规的助溶剂、缓冲剂、pH调节剂等。这些辅料是本领域常用的。
此外，如需要，也可以向药物制剂中添加着色剂、防腐剂、香料、矫味剂、甜味剂或其它材料。
为达到用药目的，增强治疗效果，本发明的药物或药物组合物可用任何公知的给药方法给药。
本发明提取物药物组合物的给药剂量取决于许多因素，例如所要预防或治疗疾病的性质和严重程度，患者或动物的性别、年龄、体重、性格及个体反应，给药途径、给药次数、治疗目的，因此本发明的治疗剂量可以有大范围的变化。一般来讲，本发明中药学成分的使用剂量是本领域技术人员公知的。可以根据本发明提取物组合物中最后的制剂中所含有的实际药物数量，加以适当的调整，以达到其治疗有效量的要求，完成本发明的预防或治疗目的。本发明提取物的每天的合适剂量范围本发明的提取物的用量为0.001～150mg/Kg体重，优选为0.1～100mg/Kg体重，更优选为1～60mg/Kg体重，最优选为2～50mg/Kg体重。上述剂量可以单一剂量形式或分成几个，例如二、三或四个剂量形式给药这受限于给药医生的临床经验以及包括运用其它治疗手段的给药方案。每一种治疗所需总剂量可分成多次或按一次剂量给药。本发明的提取物或组合物可单独服用，或与其他治疗药物或对症药物合并使用并调整剂量。
本发明通过楹树提取物对胃H+/K+-ATP酶抑制活性和对幽门结扎致大鼠胃溃疡的作用的研究，证明，楹树提取物具有显著的抗胃溃疡作用，其机理是通过抑制H+/K+-ATP酶活性。
有益技术效果
本发明楹树提取物作用物质基础和作用机理明确，制备工艺简单，适合工业化生产。
为治疗胃溃疡以及慢性胃炎的提供了一种新的药物。
附图说明
图1、楹树提取物的制备实施例1的流程图。
图2、楹树提取物对大鼠慢性醋酸型胃溃疡的保护作用。
具体实施方式
下面的实施例及药物活性实验用来进一步说明本发明，实施例仅用于说明，不能限制本发明的范围。
实施例1楹树提取物的制备方法
称取楹树茎皮粗粉1Kg，用3.5升(以每公斤药材3.5升提取溶剂计)的95％乙醇回流三次，每次3小时；提取液经减压浓缩得浸膏130g，再以20％的乙醇溶解，加到3200ml的聚酰胺(30～60目)柱上，依次用12升20％的乙醇、12升60％的乙醇进行梯度洗脱，浓缩60％的乙醇洗脱液至干，即得到楹树提取物8.0g。
药理实验
试验例1楹树提取物抑制胃H+/K+-ATP酶活性
实验方法：
H+/K+-ATP酶活性测定：将纯化的猪胃黏膜囊泡加缓冲液稀释，加药后在37℃水浴温孵20min，加入20mg/kg ATP继续温孵30min，加入10％TCA终止反应，6000rpm离心10min。取定量上清液用定磷试剂盒于660nm测定H+/K+-ATP酶水解释放的无机磷含量。计算酶活力。
结果：
数据见表1。楹树提取物(1和10mg/L)对猪胃H+/K+-ATP酶活性的抑制率分别为28％和73％。提示楹树提取物对H+/K+-ATP酶活性具有较强的抑制作用。
表1楹树提取物对猪胃H+/K+-ATP酶活性的影响
| 浓度(mg/L) | 抑制率(％) |
| 1 | 28 |
| 10 | 73 |
试验例2楹树提取物对幽门结扎致大鼠胃溃疡的保护作用
实验方法：
取禁食48小时的雄性大鼠，随机分为3组。***麻醉下行幽门结扎术，术时经十二指肠给予100和500mg/kg的楹树提取物，对照组大鼠给予生理盐水。术后20小时处死动物，取出胃，计前胃部溃疡数。结果用One-way ANOVA进行差异显著性检验。
结果：
数据见表2。楹树提取物在100和500mg/kg剂量下对幽门结扎致大鼠胃溃疡的抑制率分别为43％和86％(p＜0.05)。提示楹树提取物对幽门结扎溃疡模型有很好的保护作用。
表2楹树提取物对大鼠幽门结扎致溃疡模型的影响
| 剂量(mg/kg) | 溃疡减少率(％) |
| 100 | 43 |
| 500 | 86 |
试验例3楹树提取物对大鼠慢性醋酸型胃溃疡的保护作用
实验方法：
大鼠禁食24小时，戊巴比妥钠麻醉，腺胃部靠幽门侧注射30％醋酸20ul。动物随机分组，术后第二天开始给药，每日一次，连续10天。取出胃并固定，将胃平展于玻璃板上，用数码相机照像，图像用spot advanced软件处理，测定溃疡面积。
结果：
数据见表3。楹树提取物在100和30mg/kg剂量下对大鼠慢性醋酸型溃疡的有效率分别为49.7％和57.2％(p＜0.01)。提示楹树提取物对慢性醋酸型溃疡有很好的保护作用。
表3楹树提取物对大鼠慢性醋酸型溃疡的影响
Claims (2)
1.一种楹树(Albizia chinensis(Osbeck)Merr)提取物在制备治疗胃溃疡疾病药物中的应用，其特征在于，楹树提取物的制备方法包括如下步骤：
步骤A：粉碎楹树(Albizia chinensis(Osbeck)Merr)茎皮，以药材重量计每kg用3-5升的80-95％乙醇回流2-4次，每次2-3小时；合并提取液，提取液经减压浓缩得到楹树(Albizia chinensis(Osbeck)Merr)浸膏；
步骤B：将步骤A中获得的楹树浸膏以10-30％的乙醇溶解后，加到30～90目的聚酰胺柱上，聚酰胺干粉的体积用量为以浸膏重量计25-30升/每kg，依次用10-30％的乙醇、40-80％的乙醇进行梯度洗脱，洗脱溶剂的用量为3-5倍聚酰胺干粉的体积，浓缩40-80％的乙醇洗脱液得到所述的楹树提取物。
2.一种楹树(Albizia chinensis(Osbeck)Merr)提取物在制备H+/K+-ATP抑制剂中的应用，其特征在于，楹树提取物的制备方法包括如下步骤：
步骤A：粉碎楹树(Albizia chinensis(Osbeck)Merr)茎皮，以药材重量计每kg用3-5升的80-95％乙醇回流2-4次，每次2-3小时；合并提取液，提取液经减压浓缩得到楹树(Albizia chinensis(Osbeck)Merr)浸膏；
步骤B：将步骤A中获得的楹树浸膏以10-30％的乙醇溶解后，加到30～90目的聚酰胺柱上，聚酰胺干粉的体积用量为以浸膏重量计25-30升/每kg，依次用10-30％的乙醇、40-80％的乙醇进行梯度洗脱，洗脱溶剂的用量为3-5倍聚酰胺干粉的体积，浓缩40-80％的乙醇洗脱液得到所述的楹树提取物。
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