CN115025143B - 一种毛果鱼藤提取物的制备方法和用途 - Google Patents
一种毛果鱼藤提取物的制备方法和用途 Download PDFInfo
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Abstract
本发明公开了一种毛果鱼藤提取物的制备方法和用途，毛果鱼藤(Derris eriocarpa How)为豆科鱼藤属灌木，毛果鱼藤提取物为毛果鱼藤水提物的乙醇提取物，提取方法为：取毛果鱼藤粉碎成细粉，加入适量水加热提取若干次，收集提取液浓缩至1.0～1.5 g/ml，硅胶色谱柱对以上浓缩液进行柱层析分离，以不同浓度乙醇为流动相进行梯度洗脱，收集各洗脱物浓缩至浸膏。该毛果鱼藤提取物可以在制备治疗/预防高尿酸血症及其相关疾病或具有调节尿酸功能药物方面应用。本发明首次发现毛果鱼藤提取物具有明显的调节尿酸活性，具有制备预防或治疗高尿酸血症及其相关疾病的药物制剂的成药潜力，且药物制剂的制备方法简单，成本低，成药经济效益好。
Description
技术领域
本发明属于中医药技术领域，具体涉及一种毛果鱼藤提取物的制备方法和用途。
背景技术
高尿酸血症(HUA)为嘌呤代谢紊乱所致，是痛凤的生化基础，并与高血压、抑郁症、高脂血症、动脉粥样硬化、阿尔茨海默病、肥胖、胰岛素抵抗、肾病、炎症、感染性疾病的严重程度、肿瘤溶解综合征、更年期综合征、器官移植并发症和某些疑难病症（阻塞性睡眠呼吸暂停、原发性干燥综合征等）的发生密切相关。控制高尿酸血症对治疗痛风等严重代谢性疾病及与之相关的疾病具有重要意义。
目前，临床上用于抗高尿酸血症的药物主要为减少尿酸生成和增加尿酸排泄两方面。减少尿酸生成的药物有黄嘌呤氧化酶（Xanthine oxidase，XOD）抑制剂，主要有别嘌呤醇；fexbuxostat为黄嘌呤氧化酶/脱氢酶选择性抑制剂药，该药对适度肾损伤患者比别嘌呤醇有更好的安全性。别嘌呤醇会引起过敏、肝肾损伤及骨髓抑制等不良反应，fexbuxostat也有肝功能异常、胃肠道反应、关节痛和出疹等不良反应报道。促尿酸排泄药有丙磺舒、苯溴马隆、苯磺唑酮等，作用于肾近曲小管的尿酸盐转运体，抑制尿酸的重吸收；近来也发现苯溴马隆有引起暴发性肝炎的危险。因此，研究高效低毒的抗高尿酸血症药物已经迫在眉睫。近年研究发现中药对高尿酸血症具有一定的治疗作用，且毒副作用小。如：专利申请CN109700837A公开了高原荨麻的正丁醇提取物或醇溶部位提取物中的一种或两种的混合物具有抗高尿酸血症作用，抑制肝脏XOD活力及调控肾脏尿酸转运体基因表达的作用，可用来制备抗高尿酸血症的药物。专利CN104383292B公开了铁皮石斛总多酚富集的提取物作为制备治疗高尿酸血症药物的活性成分，可降低血液中尿酸的浓度，在治疗高尿酸血症上具有显著的效果，且不具毒副作用。专利CN103191235B公开了葛根、陈皮和松叶三种药材的混合醇提物能降低高尿酸血症的大鼠血清的UA水平，降低血清和肝的XOD活性，降低血清和肝的ADA活性。等等。可见，蛮多中药材的提取物或中药复方制剂都具有制备治疗高尿酸血症药物的成药潜力，而且，中药具有副作用小、综合治疗能力强等优点，因此，开发中药材提取物的新用途，积极探索新的治疗高尿酸血症药物的潜在药物靶标，寻找新型的治疗高尿酸血症药物是当今临床亟需解决的重要问题。
毛果鱼藤(Derris eriocarpa How)为豆科鱼藤属灌木。毛果鱼藤具有具有抗炎、利尿的作用。毛果鱼藤中含3,5-二甲氧基-4’-异丙基反式二苯乙烯、2’-deoxyisoauriculatin、flemichapparin B、1-[6-羟基-2-甲氧基-2",2"-二甲基吡喃-（5",6":3,4）]-2-（4’-甲氧基苯基）-1,2-乙二酮、1-[6-羟基-2-甲氧基-2",2"-二甲基吡喃-（5",6":3,4）]-2-（3’,4’-亚甲二氧基苯基）-1,2-乙二酮、dimethylalpinum isoflavone、4-羟基-3-[4’-0-（3,3-二甲基烯丙基）（苯基]-5-甲氧基-2",2"-二甲基吡喃-5",6":6,7）-香豆素、derrubone、甘草宁G、derrusnin、木豆素、6a-dihydroxy-9-methoxypterocarpan等成分。目前针对毛果鱼藤研究较少，主要集中在毛果鱼藤的化学成分分析方面，另外，还有一些是从毛果鱼藤中提取具有抗氧化和抗炎作用物质；如，专利CN201710529968.7公开了毛果鱼藤石油醚提取物具有抗氧化作用，特别针对II型糖尿病的氧化应激有良好效果。检测结果显示，毛果鱼藤提取物对糖尿病模型小鼠的血清氧化应激性指标SOD、GSH-PX显著性增加，亦显著性降低了MDA水平；专利CN201710529969.1公开了毛果鱼藤石油醚提取物具有抗炎作用，特别针对II型糖尿病的相关炎症有良好效果。检测结果显示，给与毛果鱼藤石油醚提取物治疗的实验模型小鼠的TNF-α、MCP-1、IL-6、IL1-β等炎症趋化因子以及炎症因子显著降低，胰腺中炎症介质因子COX-2的表达下降。但目前还没有将毛果鱼藤用于制备预防或治疗高尿酸血症及其相关疾病或具有调节尿酸功能药物的相关介绍，所以，如何将毛果鱼藤扩大化使用，使之产生最大的功效，发挥出应有的效能，仍是医学医药研究的主攻方向。
发明内容
本发明的目的是提供一种毛果鱼藤提取物的制备方法和用途，本发明首次发现毛果鱼藤提取物具有明显的调节尿酸活性，具有制备预防或治疗高尿酸血症及其相关疾病的药物制剂的成药潜力，且药物制剂的制备方法简单，成本低，成药经济效益好。
为实现上述目的，本发明采用如下技术方案：
一种毛果鱼藤提取物的制备方法，所述毛果鱼藤为为豆科鱼藤属灌木，其特征在于，所述制备方法包括以下步骤：
（1）取毛果鱼藤粉碎成细粉，加入3～9倍质量蒸馏水先浸泡30～90min，然后加热至70～100℃提取30～90min，过滤，收集滤液，保留滤渣；
（2）将步骤（1）所得滤渣重复提取1～3次，每次加入3～9倍质量蒸馏水加热至70～100℃提取30～90min，过滤，收集滤液，合并多次提取得到的滤液浓缩至1.0～1.5 g/mL，得到毛果鱼藤的水提物；
（3）采用硅胶色谱柱对毛果鱼藤的水提物进行柱层析分离，用乙醇为流动相进行梯度洗脱，收集各洗脱物浓缩至浸膏即得毛果鱼藤提取物。
步骤（3）中乙醇的体积浓度≤95%。在乙醇的体积浓度≤95%中选取若干个浓度组成梯度洗脱液。
优选地，所述乙醇的体积浓度为30%、65%、80%、95%。
以上毛果鱼藤的水提物或毛果鱼藤提取物在制备治疗/预防高尿酸血症及其相关疾病药物或具有调节尿酸功能药物方面的用途。优选地，毛果鱼藤提取物是指毛果鱼藤水提物的30～75%（体积浓度）乙醇洗脱部位。
所述高尿酸血症及其相关疾病为由尿酸代谢失调引起的高血压、高脂血症、动脉粥样硬化、肥胖、胰岛素抵抗、肾病、炎症、感染性疾病、肿瘤溶解综合征、更年期综合征、器官移植并发症、阻塞性睡眠呼吸暂停和原发性干燥综合征等中的一种或多种。
所述的毛果鱼藤提取物选择性添加常规辅料，按照常规工艺制成临床可接受的用于治疗/预防高尿酸血症及其相关疾病或具有调节尿酸功能的药物制剂。
所述制剂包括片剂、胶囊剂、颗粒剂、酊剂、酒剂等。
所述的常规辅料包括淀粉、乳糖、微晶纤维素、糊精、磷酸钙、聚乙二醇-4000、聚乙二醇-6000、羧甲基纤维素钠、羟丙纤维素或交联聚维酮等中一种以上。
本发明的有益效果是：
1、本发明首次发现毛果鱼藤提取物具有明显的调节尿酸活性，具有制备预防或治疗高尿酸血症及其相关疾病的药物制剂的成药潜力，为开发抗高尿酸血症创新药物提供了新的物质基础，具有潜在且巨大的社会效益和经济效益；
2、本发明的毛果鱼藤资源丰富，毛果鱼藤来源方便易得，可用于毛果鱼藤提取物的大量制备。虽然毛果鱼藤水提物在高用量时也具有一定的抗高尿酸血症活性，但是用量大且活性低，因此，在本发明的毛果鱼藤制备方法中，将毛果鱼藤先经过多次水提合并提取液，得到毛果鱼藤水提物，再对毛果鱼藤水提物用不同浓度的乙醇进行洗脱得到不同的洗脱部位，最后收集对高尿酸血症及其相关疾病具有显著疗效的部位，极大程度提高了最终得到的毛果鱼藤提取物及其药物制剂调节尿酸的活性；而且整个提取制备方法简单易行、成本低、污染小，利于节能减排条件下的大规模生产，产业化前景好；
3、本发明分别通过研究毛果鱼藤提取物对次黄嘌呤引起高尿酸血症小鼠血清尿酸水平的影响以及毛果鱼藤提取物对尿酸钠诱导的大鼠足趾肿胀的影响，有力证明了本发明的毛果鱼藤提取物对高尿酸血症具有明显的调节尿酸活性，毛果鱼藤水提物在高用量时也具有一定的抗高尿酸血症活性；
4、本发明的毛果鱼藤水提物或毛果鱼藤提取物还具有祛风活血等功效，可用于风湿骨痛、铁打损伤、肥大性脊柱炎等症，可以在用于预防或治疗高尿酸血症或痛风的同时起到治疗其他合并症的效果；
5、将毛果鱼藤提取物与辅料混合制成片剂、胶囊剂、滴丸、颗粒剂、酊剂、酒剂等，这些制剂较为方便服用，能单独或者搭配其他药剂服用，通过内服能将药性通过血液快速输送至病症部位进行有效治疗。
具体实施方式
为了更加详细的介绍本发明，下面结合实施例，对本发明做进一步说明。
实施例1毛果鱼藤水提物的制备
将毛果鱼藤粉碎成细粉，取毛果鱼藤细粉1kg，加入4L蒸馏水浸泡60min，然后加热至80℃提取60min，过滤，收集滤液；滤渣再加入4L蒸馏水加热至80℃提取60min，过滤，收集滤液；滤渣再加入4L蒸馏水加热至80℃提取60min，过滤，收集滤液；合并3次提取液，浓缩至1.0～1.5g/mL，即得。
实施例2 毛果鱼藤水提物的制备
将毛果鱼藤粉碎成细粉，取毛果鱼藤细粉1kg，加入6L蒸馏水浸泡60min，然后加热至90℃提取60min，过滤，收集滤液；滤渣再加入6L蒸馏水加热至90℃提取60min，过滤，收集滤液；滤渣再加入6L蒸馏水加热至90℃提取60min，过滤，收集滤液；合并3次提取液，浓缩至1.0～1.5g/mL，即得。
实施例3 毛果鱼藤水提物的制备
将毛果鱼藤粉碎成细粉，取毛果鱼藤细粉1kg，加入8L蒸馏水浸泡60min，然后加热至98℃提取60min，过滤，收集滤液；滤渣再加入8L蒸馏水加热至98℃提取60min，过滤，收集滤液；滤渣再加入8L蒸馏水加热至98℃提取60min，过滤，收集滤液；合并3次提取液，浓缩至1.0～1.5g/mL，即得。
实施例4 毛果鱼藤水提物的制备
将毛果鱼藤粉碎成细粉，取毛果鱼藤细粉1kg，加入8L蒸馏水浸泡80min，然后加热至80℃提取80min，过滤，收集滤液；滤渣再加入6L蒸馏水加热至90℃提取60min，过滤，收集滤液；滤渣再加入4L蒸馏水加热至98℃提取40min，过滤，收集滤液；合并3次提取液，浓缩至1.0～1.5g/mL，即得。
实施例5 毛果鱼藤水提物乙醇洗脱部位的制备
取采用实施例3的制备方法制备得到的毛果鱼藤水提物浓缩液，采用硅胶色谱柱进行柱层析分离，以体积浓度为5%、10%、15%、20%、25%、30%、35%、40%、45%、50%、55%、60%、65%、70%、75%、80%、85%、90%、95%的乙醇为流动相依次进行梯度洗脱，分别收集各洗脱物浓缩至浸膏即得毛果鱼藤提取物。
实施例6 毛果鱼藤水提物乙醇洗脱部位的制备
取采用实施例3的制备方法制备得到的毛果鱼藤水提物浓缩液，采用硅胶色谱柱进行柱层析分离，以体积浓度为15%、30%、45%、60%、75%、90%的乙醇为流动相依次进行梯度洗脱，分别收集各洗脱物浓缩至浸膏即得毛果鱼藤提取物。
实施例7 毛果鱼藤水提物乙醇洗脱部位的制备
取采用实施例3的制备方法制备得到的毛果鱼藤水提物浓缩液，采用硅胶色谱柱进行柱层析分离，以体积浓度为30%、60%、75%、95%的乙醇为流动相依次进行梯度洗脱，分别收集各洗脱物浓缩至浸膏即得毛果鱼藤提取物。
实施例8 毛果鱼藤水提物乙醇洗脱部位的制备
取采用实施例3的制备方法制备得到的毛果鱼藤水提物浓缩液，采用硅胶色谱柱进行柱层析分离，以体积浓度为15%、35%、55%、75%、95%的乙醇为流动相依次进行梯度洗脱，分别收集各洗脱物浓缩至浸膏即得毛果鱼藤提取物。
实施例9 毛果鱼藤提取物片剂的制备
【处方】由实施例7的方法制得的毛果鱼藤水提物的60%乙醇洗脱部位（取相当于毛果鱼藤药材4 kg的量）、干淀粉适量；
【制备方法】将毛果鱼藤水提物的60%乙醇洗脱部位、干淀粉混匀，加入适量羧甲基纤维素钠、滑石粉，制成颗粒，干燥，压制成1000片，所得的毛果鱼藤提取物片，每片含毛果鱼藤4 g。
以上所述的干淀粉、羧甲基纤维素钠和滑石粉均可在市场上购买得到。
实施例10 毛果鱼藤提取物胶囊的制备
【处方】由实施例7的方法制得的毛果鱼藤水提物的60%乙醇洗脱部位（取相当于毛果鱼藤药材4kg的量）、干淀粉适量；
【制备方法】将毛果鱼藤水提物的60%乙醇洗脱部位、干淀粉混匀，加入适量羧甲基纤维素钠、滑石粉，制成颗粒，干燥，装囊成1000粒，所得的毛果鱼藤提取物胶囊，每粒含毛果鱼藤4g。
以上所述的干淀粉、羧甲基纤维素钠和滑石粉均可在市场上购买得到。
实施例11 毛果鱼藤提取物颗粒剂的制备
【处方】由实施例7的方法制得的毛果鱼藤水提物的60%乙醇洗脱部位（取相当于毛果鱼藤药材8kg的量）、干淀粉适量；
【制备方法】将毛果鱼藤水提物的60%乙醇洗脱部位、干淀粉混匀，加入适量羧甲基纤维素钠、滑石粉，制成颗粒，干燥，分装成1000袋，所得的毛果鱼藤提取物颗粒，每袋含毛果鱼藤8g。
以上所述的干淀粉、羧甲基纤维素钠和滑石粉均可在市场上购买得到。
药性试验
为了验证本发明毛果鱼藤提取物调节尿酸的活性，本申请人进行了以下试验：
1、毛果鱼藤提取物对次黄嘌呤引起高尿酸血症小鼠血清尿酸水平的影响
供试品：所述毛果鱼藤水提物均是由实施例3中所述方法制的毛果鱼藤水提物，所述毛果鱼藤提取物均是由实施例7中所述方法制得的毛果鱼藤水提物的50%乙醇洗脱部位。
处理方法：小鼠90只，随机分为9组，每组10只，即正常组，模型组，阳性对照组（别嘌呤醇20mg.kg-1），毛果鱼藤提取物高、中、低剂量组（分别相当于毛果鱼藤药材30 g.kg-1、15 g.kg-1和7.5g.kg-1)，毛果鱼藤水提物高、中、低剂量组（分别相当于毛果鱼藤药材30g.kg-1、15 g.kg-1和7.5 g.kg-1)， 灌胃给药，每天1次，正常组和模型组灌胃给予等体积蒸馏水，连续12 d。末次给药1 h后，除正常组外，其他各组腹腔注射给予次黄嘌呤0.35 g.kg-1，2.5 h后于各组小鼠眼球取血，1000×g离心10min，取血清，采用磷钨酸比色法测定血清尿酸含量。
表1 毛果鱼藤提取物对次黄嘌呤引起高尿酸血症小鼠血清尿酸的影响（
，n=10）
与模型组比较：*P＜0.05，** P＜0.01
结果：如表1 所示，和模型组相比较，毛果鱼藤提取物高、中、低剂量组以及毛果鱼藤水提物高、中、低剂量组小鼠的血清尿算水平均明显降低，比较有统计学意义（P＜0.05）。
Claims (5)
1.一种毛果鱼藤提取物在制备治疗/预防高尿酸血症药物的用途，所述毛果鱼藤为豆科鱼藤属灌木，其特征在于，所述毛果鱼藤提取物的制备方法包括以下步骤：
（1）取毛果鱼藤粉碎成细粉，加入3～9倍质量蒸馏水先浸泡30～90min，然后加热至70～100℃提取30～90min，过滤，收集滤液，保留滤渣；
（2）将步骤（1）所得滤渣重复提取1～3次，每次加入3～9倍质量蒸馏水加热至70～100℃提取30～90min，过滤，收集滤液，合并多次提取得到的滤液浓缩至1.0～1.5 g/mL，得到毛果鱼藤的水提物；
（3）采用硅胶色谱柱对毛果鱼藤的水提物进行柱层析分离，用体积浓度为35～80%的乙醇为流动相进行梯度洗脱，收集各洗脱物浓缩至浸膏即得毛果鱼藤提取物。
2.根据权利要求1所述毛果鱼藤提取物在制备治疗/预防高尿酸血症药物的用途，其特征在于，步骤（3）中乙醇的体积浓度≤95%。
3.根据权利要求1所述毛果鱼藤提取物在制备治疗/预防高尿酸血症药物的用途，其特征在于，所述乙醇的体积浓度为30%、65%、80%、95%。
4.根据权利要求1所述毛果鱼藤提取物在制备治疗/预防高尿酸血症药物的用途，其特征在于，所述的毛果鱼藤提取物选择性添加常规辅料，按照常规工艺制成临床可接受的用于治疗/预防高尿酸血症药物。
5.根据权利要求4所述毛果鱼藤提取物在制备治疗/预防高尿酸血症药物的用途，其特征在于，所述制剂包括片剂、胶囊剂、颗粒剂、酊剂或酒剂。
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| CN105732761A (zh) * | 2016-03-08 | 2016-07-06 | 中国科学院昆明植物研究所 | 天然甜味剂齐墩果烷型三萜皂苷类化合物及其制备方法与应用 |
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| CN105732761A (zh) * | 2016-03-08 | 2016-07-06 | 中国科学院昆明植物研究所 | 天然甜味剂齐墩果烷型三萜皂苷类化合物及其制备方法与应用 |
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