CN110876702A - 红车轴草提取物用作酪氨酸酶抑制剂的用途 - Google Patents
红车轴草提取物用作酪氨酸酶抑制剂的用途 Download PDFInfo
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Abstract
本发明公开了红车轴草提取物用作酪氨酸酶抑制剂的用途。体外酶活试验结果表明：红车轴草水提取物和不同浓度乙醇提取物对酪氨酸酶均有一定的抑制活性，因此，红车轴草提取物是一种优良的酪氨酸酶天然抑制剂，有进一步开发成美容化妆品或皮肤用药的潜力。
Description
技术领域
本发明涉及红车轴草提取物的一种新用途，尤其是在用作酪氨酸酶抑制剂中的新用途。
背景技术
红车轴草(Trifolium pretense L.)为豆科蝶形花亚科车轴草属(Trifoliumlinn.)植物，为一年或多年生的草本植物，又名三叶草、红花苜蓿、红三叶、红荷兰翘摇等。红车轴草分布于世界各地，在我国主要散布于湖北、新疆、云南等地，资源较为丰富。我国民间将红车轴草花序入药，主要用于止咳、止喘以及解痉等，将其制成软膏后可治疗局部溃疡。该植物主要含异黄酮、黄酮、香豆素、有机酸等化学成分，其主要成分异黄酮具有雌激素样作用，对人体内的激素水平具有双向调节功能，从而能治疗多种与雌激素水平紊乱有关的妇科疾病如更年期综合征和骨质疏松。随着对红车轴草化学成分及药理活性研究的不断深入，还发现其对抗癌、抗疟、祛痰、解痉等有一定作用，可用于治疗百日咳及支气管炎等症。
酪氨酸酶(Tyrosinase)是一种氧化酶，是调控黑色素生成的限速酶，在黑色素生成过程中起着至关重要的作用，它可催化L-酪氨酸合成L-多巴，然后将多巴氧化生成多巴醌，进而形成色素，其在人体中过量表达是导致雀斑、黄褐斑、黑色素瘤等色素沉着的主要原因，在目前使用较多的脱色剂中，氢醌、曲酸、含汞制剂、过氧化氢等都是毒性较大的物质，故寻找天然有效的酪氨酸酶抑制剂是美白化妆品及皮肤用药的研究热点。
发明内容
本发明的一个目的是提供红车轴草提取物用作酪氨酸酶抑制剂的新用途。
体外酶活试验结果表明：红车轴草水提取物和不同浓度乙醇提取物对酪氨酸酶均有一定的抑制活性，其中乙醇提取物的活性高于水提取物，而40％乙醇提取物的活性接近于洋甘菊。
以上结果表明：红车轴草提取物是一种优良的酪氨酸酶天然抑制剂，有进一步开发成化妆品或皮肤用药的潜力。
本发明的另一个目的是提供一种酪氨酸酶抑制剂。
红车轴草提取物既可单独作为活性成分用于酪氨酸酶抑制剂，也可与其它具有相同活性的成分如洋甘菊提取物、熊果苷等一起作为活性成分用于酪氨酸酶抑制剂，从而进一步提高对酪氨酸酶的抑制活性。所述酪氨酸酶抑制剂中除了含有活性成分，还含有药学上可接受的载体。
优选地，所述红车轴草提取物是红车轴草醇提取物，所述醇包括但不限于乙醇、甲醇、丙二醇、正丁醇、异丁醇等一元或多元醇。
本发明优选地是20-80％乙醇。
本发明进一步优选地是40％乙醇。
基于以上发明，本发明进一步提供一种化妆品或日化用品，所述化妆品或日化用品中含有美白剂，所述美白剂是红车轴草提取物。所述化妆品包括面膜、乳液、霜膏等剂型，所述日化用品包括香皂、沐浴露等产品剂型。所述化妆品或日化用品的主要功能是美白。
基于以上发明，本发明再进一步提供一种治疗黑色素沉着所导致疾病的药物，该药物的活性成分是红车轴草提取物。所述黑色素沉着所导致的疾病包括但不限于雀斑、黄褐斑、黑色素瘤。
当然，本发明的用途远不局限于此，鉴于酪氨酸酶在细菌、微生物、植物、动物中都广泛存在且发挥重要作用，本发明所提供的抑制剂在农业、畜牧业、食品添加剂等领域也具有重大应用潜力，只要是与酪氨酸酶抑制剂有关的用途都在本发明的保护范围之内。
更详尽的方案请参阅具体实施例。
具体实施方式
下面通过具体实施例对本发明进行详细地说明。
实施例1红车轴草提取物的制备
取红车轴草药材，分别用水、20％、40％、60％、80％(均为体积浓度)乙醇提取，将提取液浓缩、干燥，得到红车轴草水提取物和不同浓度乙醇提取物。
实施例2红车轴草提取物抑制酪氨酸酶活性试验
将红车轴草水提取物和不同浓度乙醇提取物分别用PBS缓冲液溶解，均配制成5mg/mL的溶液，作为样品溶液。同时将洋甘菊提取物(商业途径购买)按同法制成样品溶液，作为对照。酪氨酸酶、L-多巴均购自美国Sigma公司。
将各样品溶液2mL分别与0.005mol/L的L-多巴反应液1mL混合，37℃孵育10min后，然后加入酪氨酸酶100U，PBS缓冲液将反应体系定容至4ml，37℃继续孵育10min，取出冰浴，用分光光度计于475nm处测吸光度，按以下公式计算酪氨酸酶抑制率：
抑制率I＝[1-(A-B)/(C-D)]×100％
其中，A：加入样品、L-多巴及酪氨酸酶反应后所测的吸光度；
B：加入样品、酪氨酸酶，未加入L-多巴所测的吸光度；
C：未加样品，仅加入L-多巴与酪氨酸酶反应后所测的吸光度；
D：未加样品和L-多巴，仅加入酪氨酸酶所测的吸光度。
每次试验均重复三次取平均值，结果见表1。
表1红车轴草提取物对酪氨酸酶的抑制率
| 样品种类 | 抑制率(％) |
| 红车轴草水提取物 | 39.6 |
| 红车轴草20％乙醇提取物 | 54.2 |
| 红车轴草40％乙醇提取物 | 69.6 |
| 红车轴草60％乙醇提取物 | 64.5 |
| 红车轴草80％乙醇提取物 | 58.3 |
| 洋甘菊提取物 | 73.7 |
结果分析：洋甘菊提取物是目前最常用的美容护肤美白成分，已有大量文献报道洋甘菊具有抑制酪氨酸酶的活性。从以上试验可以看出，红车轴草水提取物和不同浓度乙醇提取物对酪氨酸酶均有一定的抑制率，其中乙醇提取物的活性高于水提取物，而40％乙醇提取物的活性接近于洋甘菊。
Claims (9)
1.红车轴草提取物用作酪氨酸酶抑制剂的用途。
2.一种酪氨酸酶抑制剂，其活性成分中含有红车轴草提取物。
3.如权利要求2所述的酪氨酸酶抑制剂，其特征在于：所述红车轴草提取物是红车轴草醇提取物。
4.如权利要求3所述的酪氨酸酶抑制剂，其特征在于：所述醇是20-80％乙醇。
5.如权利要求4所述的酪氨酸酶抑制剂，其特征在于：所述醇是40％乙醇。
6.一种化妆品，含有美白剂，其特征在于：所述美白剂是红车轴草提取物。
7.一种治疗黑色素沉着所导致疾病的药物，其特征在于：该药物的活性成分是红车轴草提取物。
8.如权利要求7所述的药物，其特征在于：所述黑色素沉着所导致的疾病包括雀斑、黄褐斑、黑色素瘤。
9.如权利要求6所述的化妆品，或权利要求7或8所述的药物，其特征在于：所述红车轴草提取物是红车轴草醇提取物，尤其是20-80％乙醇提取物，最优是40％乙醇提取物。
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