CN109125376B - 一种防治旋毛虫病的药物 - Google Patents
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Abstract
本发明公开了一种防治旋毛虫病的药物，属于生物医药技术领域。本发明提供了一种获取简单、可口服给药使用方便、能够有效防治旋毛虫的杀虫药物王不留行正丁醇提取物，其中有效成分王不留行黄酮苷的含量(重量分数)不少于15％，刺桐碱的含量不少于1.5％，该药物可以使患旋毛虫病的小鼠身上成虫减少79.53％，肌肉幼虫减少77.70％。这一发现对于开发安全有效的抗旋毛虫病新药物，具有巨大的应用价值。
Description
技术领域
本发明涉及一种防治旋毛虫病的药物，属于生物医药技术领域。
背景技术
旋毛虫病是一种世界范围的严重人畜共患病，通过消化未加工的未煮熟的粪便以及含有感染性旋毛虫幼虫的其他动物的肉进行传播和感染，约有120-150种哺乳动物可自然感染此虫，尤其是猪、犬、猫、狐和某些鼠类感染率较高。旋毛虫病不仅直接危害养猪业和肉类业的经济利益，而且严重危害人类健康。
阿苯达唑(ABZ)是治疗人类旋毛虫病的基本驱虫剂。然而，其不溶于水，吸收缓慢限制了它们的吸收和生物利用度。此外，三岁以下儿童和孕妇禁用阿苯达唑。因此，目前迫切需要开发安全有效的抗旋毛虫病新药物，特别是那些副作用小，耐受性高的中药提取物。
发明内容
本发明的第一个目的是提供王不留行正丁醇提取物在制备防治旋毛虫病的药物中的应用，所述王不留行正丁醇提取物中王不留行黄酮苷的含量不少于15％，王不留行刺桐碱的含量不少于1.5％。
在本发明的一种实施方式中，所述药物中包含王不留行正丁醇提取物、药物载体和/或药用辅料。
在本发明的一种实施方式中，所述药物载体包含微囊、微球、纳米粒以及脂质体。
在本发明的一种实施方式中，所述药用辅料包含赋形剂以及附加剂。
在本发明的一种实施方式中，所述药用辅料包含溶剂、抛射剂、增溶剂、助溶剂、乳化剂、着色剂、黏合剂、崩解剂、填充剂、润滑剂、润湿剂、渗透压调节剂、稳定剂、助流剂、矫味剂、防腐剂、助悬剂、包衣材料、芳香剂、抗黏合剂、整合剂、渗透促进剂、pH值调节剂、缓冲剂、增塑剂、表面活性剂、发泡剂、消泡剂、增稠剂、包合剂、保湿剂、吸收剂、稀释剂、絮凝剂与反絮凝剂、助滤剂以及释放阻滞剂。
在本发明的一种实施方式中，所述附加剂包含微晶纤维素、羟丙基甲基纤维素以及精制卵磷脂。
在本发明的一种实施方式中，所述药物的剂型包括颗粒剂、胶囊剂、片剂、丸剂和口服液。
在本发明的一种实施方式中，所述王不留行正丁醇提取物应用的产品形式包含药物制剂、食品、营养品以及保健品。
本发明的第二个目的是提供一种防治旋毛虫病的药物，所述药物中包含王不留行正丁醇提取物，所述王不留行正丁醇提取物中王不留行黄酮苷的含量不少于15％，王不留行刺桐碱的含量不少于1.5％。
本发明的第三个目的是提供上述药物的制备方法，所述方法包括：称取炒王不留行种子，粉碎后经石油醚脱脂，然后用10倍(质量)的70％乙醇85℃水浴加热回流提取2h，减压浓缩得70％乙醇提取物，待挥发至无乙醇气味后，混悬于水中依次用乙酸乙酯和水饱和正丁醇萃取2次；合并水饱和正丁醇相，减压浓缩至干，去离子水复溶，真空干燥得王不留行正丁醇相干粉，经HPLC检测其中王不留行黄酮苷的含量(重量分数)不低于15％，刺桐碱的含量不低于1.5％。
本发明的有益效果：
本发明通过制备王不留行黄酮苷的含量不少于15％且王不留行刺桐碱的含量不少于1.5％的王不留行正丁醇提取物，并将其应用于感染旋毛虫的小鼠，通过喂食，发现王不留行正丁醇提取物可使成虫减少79.53％，肌肉幼虫减少77.70％。而且，该王不留行正丁醇提取物还具备一定的水溶性，比旋毛虫病的常用药物阿苯达唑(ABZ)更易被吸收利用。这一发现对于开发安全有效的抗旋毛虫病新药物，具有巨大的应用价值。
具体实施方式
(一)HPLC检测王不留行黄酮苷和刺桐碱含量的方法：
将正丁醇提取物干粉溶解于甲醇中备用。使用色谱柱AgilentC18柱(4.6mm×250mm,5μm)；流动相使用甲醇-0.3％磷酸溶液；流速设为0.8mL/min；检测波长设为280nm；柱温设为30℃；进样量设为15μL。依次检测王不留行黄酮苷和刺桐碱标准品，然后检测正丁醇提取物干粉中王不留行黄酮苷和刺桐碱的含量。
实施例1：王不留行正丁醇提取物的制备
取100g炒王不留行种子，粉碎后经石油醚脱脂。然后用10倍(质量)的70％乙醇85℃水浴加热回流提取2h，减压浓缩得70％乙醇提取物，待挥发至无乙醇气味后，混悬于水中依次用乙酸乙酯和水饱和正丁醇萃取2次。合并水饱和正丁醇相，减压浓缩至干，去离子水复溶，真空干燥得王不留行正丁醇相干粉，经HPLC检测其中王不留行黄酮苷的含量(重量分数)不低于15％，刺桐碱的含量不低于1.5％。
实施例2：王不留行正丁醇提取物的杀虫药物药效评估
用300只旋毛虫幼虫口服感染6周龄ICR实验小鼠，感染旋毛虫的小鼠随机分为3组：Ⅰ组：感染非治疗组(即对照组，n＝12，n为小鼠数量)；Ⅱ组(肌肉期，n＝12)：从感染后第7天开始口服给药王不留行正丁醇提取物，并持续14天；Ⅲ组(肌肉期，n＝12)：从第7天开始接受ABZ给药并持续14天(Ⅱ组、Ⅲ组的用药量相同)。分别在第7天和第40天通过颈脱位处死小鼠6只。获得肌肉样品对旋毛虫成虫和幼虫进行计数统计。结果如表1所示。
表1王不留行正丁醇提取物的杀虫药物药效评估
在小鼠被感染后第7天和第40天我们对旋毛虫成虫和肌肉中幼虫的寄生虫数量进行了统计。结果表明，与对照组相比，王不留行正丁醇提取物和ABZ处理组成虫分别下降79.53％和81.22％，肌肉幼虫减少77.70％和90.70％。因此，王不留行正丁醇提取物是一种有效的用于防治旋毛虫病的药物，并且可能还是一种潜在的用于治疗由旋毛虫感染引起的炎症和相关疾病的有效药物。
另外，在20℃下我们测定了所得王不留行正丁醇提取物在100g水中达到饱和状态时所溶解的质量3.43g，即溶解度为3.43g。与阿苯达唑相比，本发明所提供的王不留行正丁醇提取物溶解度从0.3g提高到3.43g。这说明王不留行正丁醇提取物比阿苯达唑更易吸收，生物利用度更高，且王不留行正丁醇提取物作为中药提取物，其应用范围更广。
对比例1：王不留行黄酮苷和刺桐碱的杀虫药物药效评估
用300只旋毛虫幼虫口服感染6周龄ICR实验小鼠，感染旋毛虫的小鼠随机分为3组：Ⅳ组(肌肉期，n＝12)：从感染后第7天开始口服从王不留行正丁醇提取物中分离到的王不留行黄酮苷，并持续14天；Ⅴ组(肌肉期，n＝12)：从第7天开始口服从王不留行正丁醇提取物中分离到的王不留行刺桐碱并持续14天；Ⅵ组(肌肉期，n＝12)：从第7天开始口服市售王不留行黄酮苷和市售王不留行刺桐碱的复合物(按照王不留行黄酮苷与刺桐碱质量比为10:1进行复配)并持续14天(Ⅱ组、Ⅲ、Ⅳ、Ⅴ、Ⅵ组的用药质量相同)。分别在第7天和第40天通过颈脱位处死小鼠6只。获得肌肉样品对旋毛虫成虫和幼虫进行计数统计，如表2所示。
结果表明，仅口服从王不留行正丁醇提取物中分离到的王不留行黄酮苷、仅口服从王不留行正丁醇提取物中分离到的王不留行刺桐碱、口服市售王不留行黄酮苷和市售王不留行刺桐碱质量比为10:1进行复配得到的复合物时，与对照组(Ⅰ组)相比，小鼠身上的旋毛虫成虫数量分别下降了18.27％、55.32％和54.31％，小鼠身上的旋毛虫幼虫数量分别下降了12.88％、37.56％和38.95％。
表2王不留行黄酮苷和/或刺桐碱的杀虫药物药效评估
虽然本发明已以较佳实施例公开如上，但其并非用以限定本发明，任何熟悉此技术的人，在不脱离本发明的精神和范围内，都可做各种的改动与修饰，因此本发明的保护范围应该以权利要求书所界定的为准。
Claims (5)
1.王不留行正丁醇提取物在制备防治旋毛虫病的药物中的应用，其特征在于，所述王不留行正丁醇提取物中包括含量不少于15%的王不留行黄酮苷和含量不少于1.5%的王不留行刺桐碱；
所述王不留行正丁醇提取物的制备方法，包括：
（1）称炒王不留行种子，粉碎，脱脂；然后用10倍质量的70%乙醇，85℃加热回流提取2h，减压浓缩得70%乙醇提取物；
（2）待挥发至无乙醇气味后，混悬于水中依次用乙酸乙酯和水饱和正丁醇萃取2次，合并水饱和正丁醇相，减压浓缩至干，复溶，干燥，即得王不留行正丁醇相干粉。
2.根据权利要求1所述的应用，其特征在于，所述药物中包含王不留行正丁醇提取物、药物载体和/或药用辅料；所述药物载体包含微囊、微球、纳米粒以及脂质体；所述药用辅料包含赋形剂以及附加剂。
3.根据权利要求2所述的应用，其特征在于，所述附加剂包含微晶纤维素、羟丙基甲基纤维素以及精制卵磷脂。
4.根据权利要求1-3任一所述的应用，其特征在于，所述药物的剂型包括颗粒剂、胶囊剂、片剂、丸剂和口服液。
5.根据权利要求4所述的应用，其特征在于，所述王不留行正丁醇提取物应用的产品形式为药物制剂。
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