CN103720723A - 一种绵马贯众水提物及其制备方法和应用 - Google Patents
一种绵马贯众水提物及其制备方法和应用 Download PDFInfo
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- CN103720723A CN103720723A CN201210390422.5A CN201210390422A CN103720723A CN 103720723 A CN103720723 A CN 103720723A CN 201210390422 A CN201210390422 A CN 201210390422A CN 103720723 A CN103720723 A CN 103720723A
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- Prior art keywords
- rhizoma dryopteris
- dryopteris crassirhizomatis
- water extract
- residue
- rhizoma
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Abstract
本发明涉及一种绵马贯众的水提物及其制备方法和应用,该水提物由以下方法制备:用水煎煮,煎煮液浓缩,即得。或采用以下方法制备:绵马贯众粉碎,用石油醚浸泡后,渗滤提取,回收残渣,残渣用乙醇提取,得到残渣和提取液,其中残渣用水煎煮,煎煮液浓缩,得到绵马贯众水提物。本发明提供的绵马贯众水提物对老年痴呆有防治作用。
Description
技术领域
本发明涉及绵马贯众的提取物,具体涉及一种绵马贯众水提物及其制备方法和应用。
背景技术
老年痴呆症是严重威胁人类健康的神经退行性疾病。Alzheimerdisease(简称AD)是一种主要老年痴呆症,表现为记忆、理解、计算、判断、定向、语言、抽象思维、自我控制等能力障碍,导致患者独立生活和工作能力丧失。随着社会老龄化的加速,AD已成为第4大引起人类死亡的疾病,我国即将进入老龄社会,而且该病逐渐呈现的年轻化趋势,发病率在逐年增高。迄今为止尚未有治愈老年痴呆症的有效手段,但服用乙酰胆碱酯酶抑制剂(ACHEI)是目前治疗老年痴呆症最有效的治疗方式。临床上使用的多奈哌齐(donepezil),美曲膦酯(metrifonate)和石杉碱甲(huperzine A)等药物普遍存在外周副作用较大,长期治疗效果不理想的现象。新研究表明,给予大鼠大脑皮层内灌注选择性丁酰胆碱酶抑制剂(BuCHEI)可使乙酰胆碱浓度增加15倍,且未发现胆碱样副作用。临床试验也表明,脑脊液中BuCHE抑制与AD患者的认知功能具有相关性。因此这为寻找疗效AD病,具有成本低廉、效果好,毒副作用小的活性化合物提供了新的方法。
绵马贯众又名:贯节、贯渠、百头、虎卷、扁苻、贯来、贯中、渠母、贯钟、伯芹、药渠、黄钟、伯萍、乐藻、草鸱头、伯药、药藻、凤尾草、蕨薇菜根、贯仲、管仲、东北绵马贯众。是鳞毛蕨科植物粗茎鳞毛蕨的Dryopteris crassirhizoma Nakai.的干燥根茎及叶柄残基、具有清热解毒、止血杀虫之功效。绵马贯众的化学成分主要有异槲皮苷、紫云英苷、冷蕨苷、绵马贯众苷、绵马贯众素、杜鹃素、绵马酚、绵马次酚、茶烯-b、铁线蕨酮等;间苯三酚类化合物包括黄绵马酸BB、PB、AB,白绵马素、新绵马素等。
现代药理研究证实,绵马贯众具有以下作用:
1、抗菌作用:实验证实:绵马贯众水提液对痢疾杆菌、伤寒杆菌、大肠杆菌、变形杆菌、绿浓杆菌、柘草芽胞杆菌、金黄色葡萄球菌及部分皮肤真菌均有不同抑制作用。
2、驱虫作用:绵马贯众对动物血吸虫病的实验治疗有显著疗效,有抗日本血吸虫作用。对猪蛔虫、绵羊肺线虫。肝吸虫的活动有不同程度的抑制,能驱除牛肝蛭等。
3、抗病毒作用:研究表明,绵马贯众不仅在体外,而且在体内对白血病细胞有明显的抑制作用;美国加州大学张以满和香港中文大学杨显荣发现狗脊具有抑制艾滋病毒的作用;绵马贯众水、乙醇两种提取物对流感病毒(FMl株)、呼吸道合胞病毒(Long)、副流感病毒(I型、Ⅲ型)、腺病毒(AD3)均具有一定的抗病毒作用。
4、对子宫平滑肌的作用:绵马贯众对家兔离体及在体子宫平滑肌有显著的兴奋作用,收缩增强,张力提高。
5、止血作用:炒炭后止血作用增强,出血时闻和凝血时间比生品明显缩短。
6、抗白血病作用:研究采用体外甲基纤维素培养法与L一651自血病小鼠体内实验法,观察砒霜、斑蝥、绵马贯众和牵牛子等10味中药的抗白血病作用.并用柔红霉素和环磷酰胺作了对比研究,结果表明中药不仅在体外,而且在体内对自血病细胞具有明显的抑制作用,可明显延长L一65l小鼠生存期。
7、抗衰老作用:潘伟槐等用不同浓度的绵马贯众水提物的培养基饲养果蝇,测定不同日龄果蝇体内SOD活性及MDA含量的变化,结果与对照组比较,绵马贯众能够显著提高果蝇体内SOD活性,抑制体内MDA含量增加,从而起到延缓衰老的功效。绵马贯众可以明显提高果蝇的平均寿命。
关于绵马贯众的提取物,有以下相关报道:
CN97112454.X公开了绵马贯众有效部位在制备抗疟疾的应用,其中其有效部位的制备方法为:绵马贯众粉碎,用乙醚回流提取,浓缩,室温放置1-2周,析出黄褐色沉淀,凝集沉淀,用乙醚充分溶解后搅拌均匀,抽滤,至溶液为淡黄色为止,沉淀自然干燥,得到浅黄色粉末,即得。
CN03150044.7公开了绵马贯众有效部位及单体的新制药用途,具体说是绵马贯众有效部位及单体在制备抗癌和抗关节炎药物中的应用,该绵马贯众有效部位的提取方法有两个:第一个是,绵马贯众粉碎,用甲醇、丙酮或乙醚回流提取,提取液浓缩,室温放置,其余方法同上一个专利的方法;第二个方法是,绵马贯众用二氧化碳超临界萃取,提取温度35-65℃、提取压力20-30MPa。
CN200910162507.6涉及一种适用于防治果树、蔬菜等经济作物上害螨的植物杀螨剂及其制备方法和用途,其公开了绵马贯众的提取物为绵马贯众与乙醇按照1:2-8的比例,温浸4-24h,反复渗漉得到,其中萃取剂可以为石油醚。
CN200910045493.X公开了绵马贯众的醇提物,其制备方法包括以下步骤:绵马贯众用95%乙醇冷浸提取,滤过,药渣于室温下置于通风处晾干,然后用热水提取3次,滤过,合并提取液,浓缩,离心,上清液及三氯醋酸去游离蛋白,离心,上清液用水透析3天,透析液浓缩至小体积后,加乙醇至含醇量为80%,离心,沉淀,冷冻干燥,即得绵马贯众的总多糖提取物。可以用于防治系统性红斑狼疮、急性呼吸窘迫综合症,
未见到绵马贯众的石油醚浸提方面的报道及其在抗老年痴呆方面的报道。
发明内容
本发明的目的是提供一种绵马贯众的提取物。
本发明提供的绵马贯众提取物为绵马贯众水提物。
本发明提供的绵马贯众水提物由以下方法制备:绵马贯众用水煎煮,煎煮液浓缩,得到绵马贯众的水提物。
优选地,所述绵马贯众水提物由以下方法制备:绵马贯众用6-10倍量的水煎煮1-3次,每次煎煮1-3h,煎煮液浓缩,得到绵马贯众的水提物。
进一步优选,所述绵马贯众的水提物由以下方法制备:绵马贯众用水煎煮2次,每次加入药材重量的6倍量的水煎煮,每次煎煮2小时,合并两次煎煮液,浓缩至干,得到绵马贯众水提物。
上述水提物中:
所述绵马贯众的使用部位为根茎及叶柄残基。
本发明还提供了上述绵马贯众水提物的制备方法,该方法包括以下步骤:绵马贯众用6-10倍量的水煎煮1-3次,每次1-3h,煎煮液浓缩,得到绵马贯众的水提物。
本发明还提供了含上述绵马贯众的水提物的制剂,由绵马贯众的水提物和药学上可接受的载体或稀释剂组成。
本发明提供的一种绵马贯众水提物,由以下方法制备:绵马贯众粉碎,用石油醚浸泡后,渗滤提取,回收残渣,残渣用乙醇提取,得到残渣和提取液,其中残渣用水煎煮,煎煮液浓缩,得到绵马贯众的水提物。
优选地,所述绵马贯众水提物,由以下方法制备:
绵马贯众粉碎,用石油醚浸泡18-36h后,渗滤提取至3-8倍药材量的渗滤液,回收残渣,残渣用70%-90%乙醇提取2-3次,每次提取1-3小时,得到残渣和提取液,残渣用水煎煮,煎煮1-3次,每次1-3h,煎煮液浓缩,得到绵马贯众的水提物。
进一步优选,绵马贯众的水提物,由以下方法制备:
1)绵马贯众粉碎,用石油醚浸泡24-36h后,渗滤提取至5-6倍药材量的渗漉液,回收残渣;
2)将步骤1)得到的残渣挥干石油醚,用3-8倍量体积浓度为70-80%的乙醇提取2次,每次1.5-2小时,得到残渣和提取液;
3)将步骤2)得到的残渣挥干乙醇,用绵马贯众6-8倍量的水煎煮2-3次,每次1.5-2小时,合并两次煎煮液,浓缩,至干,得到绵马贯众水提物。
上述水提物中:
所述绵马贯众优选为绵马贯众,所述绵马贯众的使用部位为根茎及叶柄残基。
所述粉碎为250-850μm,优选为350-710μm的粒径;
所述石油醚为60-90℃的石油醚,其用量为药材重量的的3-5倍量体积,以绵马贯众为100g为例,则石油醚的用量为300-500ml。
本发明还提供了绵马贯众水提物的另一个制备方法,该方法包括以下步骤:绵马贯众粉碎,用石油醚浸泡后,渗滤提取,回收残渣,残渣用乙醇提取,得到残渣和提取液,其中残渣用水煎煮,煎煮液浓缩,得到绵马贯众的水提物。
本发明还提供了含上述绵马贯众水提物的制剂,由绵马贯众的水提物和药学上可接受的载体或稀释剂组成。
所述药学上可接受的载体或稀释剂是指药学领域常规的药物载体,选自填充剂、粘合剂、崩解剂、润滑剂、助悬剂、润湿剂、溶剂、表面活性剂或矫味剂中的一种或几种。
所述填充剂选自淀粉、蔗糖、乳糖、甘露醇、山梨醇、木糖醇、微晶纤维素或葡萄糖等;
所述粘合剂选自纤维素衍生物、藻酸盐、淀粉、糊精、明胶或聚乙烯吡咯烷酮等;
所述崩解剂选自微晶纤维素、羧甲基淀粉钠、交联聚乙烯吡咯烷酮、低取代羟丙基纤维素或交联羧甲基纤维素钠;
所述润滑剂选自硬脂酸、聚乙二醇、碳酸钙、碳酸氢钠、微粉硅胶、滑石粉或硬脂酸镁;
所述助悬剂选自微粉硅胶、蜂蜡、纤维素、固态聚乙二醇;
所述润湿剂选自甘油、吐温-80、乙氧基氢化蓖麻油或卵磷脂;
所述溶剂选自乙醇、液态聚乙二醇、异丙醇、吐温-80、甘油、丙二醇或植物油,所述植物油选自大豆油、蓖麻油、花生油、调和油等;
所述表面活性剂选自十二烷基苯磺酸钠、硬脂酸、聚氧乙烯-聚氧丙烯共聚物、脂肪酸山梨坦或聚山梨酯(吐温)等;
所述矫味剂选自阿斯巴甜、蔗糖素、香精、柠檬酸或糖精钠。
所述制剂为片剂、糖衣丸、糖浆、合剂、胶囊、注射溶液、溶液、乳液、油膏、乳膏、吸入剂、气雾剂或栓剂。
本发明还提供了绵马贯众的水提物或含绵马贯众的水提物的制剂在制备防治老年痴呆的药物中的应用。
本发明提供的绵马贯众提取物具有以下优点:
本发明提供的绵马贯众的水提物对AD(老年痴呆)有防治作用,这一提取部位能显著抑制AD模型中丁酰胆碱酯酶的活性;绵马贯众对AD模型动物能改善学习记忆行为,因此,绵马贯众的水提物能用于制备预防和治疗老年痴呆症的药物。
具体实施方式
以下实施例用于说明本发明,但不用来限制本发明的范围。
实施例1:绵马贯众的水提物
1)取绵马贯众的根茎和叶柄残基,粉碎,过35目筛,粒径500μm;
2)取绵马贯众粗粉500克,用2000mL石油醚(60-90℃)浸泡24h后,渗滤提取,至提取液颜色浅(相当于5倍药材量),回收残渣;
3)挥干步骤2)提供的残渣中的石油醚,用残渣体积3倍量的80%乙醇提取2次,每次2小时,分别收集残渣和提取液;
4)将步骤3)得到的残渣用绵马贯众重量的6倍量的蒸馏水煎煮,煎煮2次,每次2h,合并两次水煎液,浓缩至干,得绵马贯众水提物56.52g(收率是11.30%)。
实施例2:绵马贯众的水提物
1)取绵马贯众的根茎和叶柄残基,粉碎,过60目筛,粒径250μm;
2)取绵马贯众粗粉500克,用2000mL的石油醚(60-90℃)浸泡24h后,渗滤提取,至提取液颜色浅(相当于6倍药材量),回收残渣;
3)挥干步骤2)提供的残渣中的石油醚,用残渣体积5倍量的70%乙醇提取2次,每次2小时,分别收集残渣和提取液;
4)将步骤3)得到的残渣用绵马贯众重量的8倍量的蒸馏水煎煮,煎煮3次,时间分别为2h、1.5h、1.5h,合并三次水煎液,浓缩至干,得绵马贯众水提物49.46g(收率是9.89%)。
实施例3:绵马贯众的水提物
1)取绵马贯众的根茎和叶柄残基,粉碎,过25目,粒径为710μm;
2)取绵马贯众粗粉500克,用2000mL的石油醚(60-90℃)浸泡36h后,渗滤提取,至提取液颜色浅(相当于6倍药材量),回收残渣;
3)挥干步骤2)提供的残渣中的石油醚,用残渣体积6倍量的90%的乙醇提取3次,提取时间分别为2小时、1.5小时、1.5小时,分别收集残渣和提取液;
4)将步骤3)得到的残渣用绵马贯众重量的6倍量的蒸馏水煎煮,煎煮2次,煎煮时间分别为2h、1.5h,合并两次水煎液,浓缩至干,得绵马贯众水提物63.59g(收率是12.72%)。
实施例4:绵马贯众的水提物
1)取绵马贯众的根茎和叶柄残基,粉碎,过45目筛,粒径350μm;
2)取绵马贯众粗粉500克,用1500mL石油醚(60-90℃)浸泡24h后,渗滤提取,至提取液颜色浅(相当于5倍药材量),回收残渣;
3)挥干步骤2)提供的残渣中的石油醚,用残渣体积8倍量的70%乙醇提取2次,每次2小时,分别收集残渣和提取液;
4)将步骤3)得到的残渣用绵马贯众重量的6倍量的蒸馏水煎煮,煎煮2次,每次2h,合并两次水煎液,浓缩至干,得绵马贯众水提物50.72g(收率是10.14%)。
实施例5:绵马贯众的水提物
1)取绵马贯众的根茎和叶柄残基,粉碎,过45目筛,粒径350μm;
2)取绵马贯众粗粉500克,用2500mL石油醚(60-90℃)浸泡24h后,渗滤提取,至提取液颜色浅(相当于5倍药材量),回收残渣;
3)挥干步骤2)提供的残渣中的石油醚,用残渣体积6倍量的80%乙醇提取2次,每次2小时,分别收集残渣和提取液;
4)将步骤3)得到的残渣用绵马贯众重量的6倍量的蒸馏水煎煮,煎煮2次,每次2h,合并两次水煎液,浓缩至干,得绵马贯众水提物59.57g(收率是11.91%)。
实施例6:绵马贯众的水提物
1)取绵马贯众的根茎和叶柄残基,粉碎,过35目筛,粒径500μm;
2)取绵马贯众粗粉500g,用药材重量的6倍量的蒸馏水煎煮,煎煮2次,每次2h,合并两次水煎液,浓缩至干,得绵马贯众水提物59.35g(收率是11.87%)。
实施例7:绵马贯众水提物的合剂
1、组成:绵马贯众的水提物(实施例1的方法制备)5g,水125mL,山梨酸钾3.5g。
2、制备方法:绵马贯众水提物5g,加125mL蒸馏水溶解,过滤后加入山梨酸钾3.5g,混合均匀即得。
实施例8:含绵马贯众水提物的糖浆剂
1、组成:绵马贯众的水提物(实施例2的方法制备)5g,水45mL,单糖浆50mL,甘油10mL
2、制备方法:按照配比称取原辅料,混合均匀,即得。
实施例9:含绵马贯众水提物的胶囊
1、组成:绵马贯众的水提物(实施例6的方法制备)50g,淀粉15g,硬脂酸镁5g。
2、制备方法:按照配比称取原辅料,混合均匀,分装胶囊。
实施例10:绵马贯众水提物的颗粒剂
1、组成:绵马贯众的水提物(实施例3的方法制备)50g,蔗糖粉20g。
2、制备方法:按照配比称取原辅料,混合均匀,制成颗粒。
实验例1:绵马贯众的活性部位对丁酰胆碱酯酶抑制活性的测定
1、实验原理
丁酰胆碱酯酶(BuChE)水解丁酰胆碱(BuCh)生成胆碱及丁酸,胆碱可以与巯基显色剂5,5-二硫双硝基苯甲酸(DTNB)反应生成TNB黄色化合物,根据颜色深浅进行比色定量,水解产物胆碱的数量可以反应BuChE的活力。
2、实验分组
样品组:自制备,其中绵马贯众的水提物的制备方法参照实施例1和实施例6的方法制备,绵马贯众的石油醚提取物、乙醇提取物和乙酸乙酯提取物的制备方法具体如下;
绵马贯众的石油醚提取物:取绵马贯众粉碎,过二号筛,先用石油醚浸泡24h后,渗滤提取至5倍药材量,渗漉液减压浓缩,即得;
绵马贯众的乙醇提取物和乙酸乙酯提取物,分别为:
取绵马贯众粉碎,过二号筛,先用石油醚浸泡24h后,渗滤提取至5倍药材量,回收残渣,挥至干残渣中的石油醚,用3倍量体积浓度为80%的乙醇提取2次,每次2小时,合并两次浓缩液,减压浓缩至无醇味,用浓缩液等量的乙酸乙酯萃取,倒出上层,即得乙酸乙酯提取物,放出下层,即得乙醇提取物。
阳性对照药:四异丙基甲基焦磷酰胺(Iso-OMPA),购自SIGMA公司;
3、实验方法:
以0.05mol·L-1磷酸盐缓冲液稀释人血浆80倍的稀释液作为酶反应液,3mmol·L-1作为底物的终浓度,DTNB试液的浓度为0.625mmol·L-1,终浓度0.25mmol·L-1;在酶反应体系中,50μg·L-1的待测化合物(即本发明的样品)与BuChE悬浮于反应缓冲液中(pH7.2),加入定量的底物BuCh及显色剂DTNB,37°C孵育60分钟后,在波长412nm测定吸光值。反应体系的吸光值可以反映体系中消耗BuCh的量。根据吸光值的改变可以计算化合物对BuChE活性的抑制率。
4、统计方法:抑制率计算:抑制率/%=(A标准-A样品)/(A标准-A空白)×100。
5、实验结果:见表1
表1:绵马贯众的各提取部位对丁酰胆碱酯酶活力的抑制作用
组别 | 浓度 | 抑制率% | IC50(ug/mL) |
四异丙基甲基焦磷酰胺(ISO-OMPA) | 50 | 94.43 | 8.95 |
绵马贯众的石油醚提取物 | 50 | 47.27-- | |
绵马贯众的乙酸乙酯提取物 | 50 | 38.65-- | |
绵马贯众的乙醇提取物 | 50 | 44.12-- | |
绵马贯众的水提物(实施例1) | 50 | 63.12 | 42.027 |
绵马贯众的水提物(实施例6) | 50 | 60.24 | 25.42 |
表1结果显示:本发明提供的绵马贯众的水提物有较强的丁酰胆碱酯酶抑制作用。
同理,对本发明的提取实施例和提取物进行实验,采用石油醚提取的绵马贯众提取物的效果同实施例1;采用水提的绵马贯众提取物同实施例6。
虽然,上文中已经用一般性说明、具体实施方式及试验,对本发明作了详尽的描述,但在本发明基础上,可以对之作一些修改或改进,这对本领域技术人员而言是显而易见的。因此,在不偏离本发明精神的基础上所做的这些修改或改进,均属于本发明要求保护的范围。
Claims (10)
1.一种绵马贯众水提物,其特征在于,所述绵马贯众水提物由以下方法制备:绵马贯众用水煎煮,煎煮液浓缩,得到绵马贯众水提物。
2.根据权利要求1所述的绵马贯众水提物,其特征在于,该绵马贯众水提物由以下方法制备:绵马贯众用6-10倍量的水煎煮1-3次,每次煎煮1-3h,煎煮液浓缩,得到绵马贯众水提物。
3.根据权利要求1所述的绵马贯众水提物,其特征在于,所述绵马贯众的水提物由以下方法制备:绵马贯众用水煎煮2次,每次加入药材重量的6倍量的水煎煮,每次煎煮2小时,合并两次煎煮液,浓缩,至干,得到绵马贯众水提物。
4.含权利要求1-3任一项所述的绵马贯众水提物的制剂,其特征在于,所述制剂由绵马贯众水提物和药学上可接受的载体或稀释剂组成。
5.一种绵马贯众水提物,其特征在于,所述绵马贯众的水提物由以下方法制备:绵马贯众粉碎,用石油醚浸泡后,渗滤提取,回收残渣,残渣用乙醇提取,得到残渣和提取液,其中残渣用水煎煮,煎煮液浓缩,得到绵马贯众水提物。
6.根据权利要求5所述的绵马贯众水提物,其特征在于,所述绵马贯众的水提物由以下方法制备:
绵马贯众粉碎,用石油醚浸泡18-36h后,渗滤提取至3-8倍药材量的渗滤液,回收残渣,残渣用70%-90%乙醇提取2-3次,每次提取1-3小时,得到残渣和提取液,残渣用水煎煮,煎煮1-3次,每次1-3h,煎煮液浓缩,得到绵马贯众水提物。
7.根据权利要求5所述的绵马贯众水提物,其特征在于,所述绵马贯众的水提物由以下方法制备:
1)绵马贯众粉碎,用石油醚浸泡24-36h后,渗滤提取至5-6倍药材量的渗漉液,回收残渣;
2)将步骤1)得到的残渣挥干石油醚,用3-8倍量体积浓度为70-80%的乙醇提取2次,每次1.5-2小时,得到残渣和提取液;
3)将步骤2)得到的残渣挥干乙醇,用绵马贯众6-8倍量的水煎煮2次,每次1.5-2小时,合并两次煎煮液,浓缩至干,得到绵马贯众水提物。
8.根据权利要求5-7任一项所述的绵马贯众水提物,所述粉碎为250-850μm,优选350-70μm的粒径;所述石油醚为60-90℃的石油醚,其用量为药材重量的的3-5倍量体积。
9.含权利要求5-8任一项所述的绵马贯众水提物的制剂,其特征在于,所述制剂由绵马贯众水提物和药学上可接受的载体或稀释剂组成。
10.权利要求1-8任一项所述的绵马贯众水提物或权利要求9所述的制剂在制备防治老年痴呆的药物中的应用。
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CN109939100B (zh) * | 2019-04-22 | 2022-04-19 | 井冈山大学 | 利用密羽贯众制备的乙酰胆碱酶抑制剂、制备方法及应用 |
CN109908120B (zh) * | 2019-04-22 | 2022-04-19 | 井冈山大学 | 一种狭顶鳞毛蕨有效成分、提取方法及其应用 |
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