CN105407871A - 护肤组合物 - Google Patents
护肤组合物 Download PDFInfo
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- CN105407871A CN105407871A CN201480043974.6A CN201480043974A CN105407871A CN 105407871 A CN105407871 A CN 105407871A CN 201480043974 A CN201480043974 A CN 201480043974A CN 105407871 A CN105407871 A CN 105407871A
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- extract
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- skin
- cell
- ore deposit
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Abstract
本发明涉及一种组合物,当将其施用于皮肤或摄入时,其提供增强的大/微循环。当局部施用时,该组合物具有在诸如减少黑眼圈、头皮健康、减少腋下发黑的领域的应用,且当口服摄入时,其提供了心血管疾病减少。使用茜草(<i>Rubia?cordifolia</i>)和紫矿(<i>Butea?monosperma</i>)提取物的组合实现了抗炎症和包括抗衰老和皮肤美白益处的皮肤健康益处。
Description
发明领域
本发明涉及一种组合物,当将其施用在皮肤上或摄入时,会形成增强的血液大循环或血液微循环。当局部施用时,该组合物可应用于以下领域:诸如减少黑眼圈、头皮健康、减少腋下发黑,而当口服摄入时,其可减少心血管疾病。在这两种情况下,均存在抗炎的益处以及其它皮肤健康的益处,包括抗衰老和皮肤美白。
发明背景
生活方式的改变与年龄使得发生重要健康问题(如心血管危险因素)的可能性增加。血管弹性的损伤是发生此类疾病的主要危险因素,其反映为高血压、心绞痛和动脉粥样硬化。在人类中,对内皮功能的研究已成为大循环或微脉管系统的重点。微循环是将新鲜血液递送到存在于嵌入器官组织内的脉管系统中的最细小的血管。这与大循环截然不同,后者将血液输送至器官并从器官运出。 微循环的主要功能包括:1.调节血液流动和组织灌注;2.调节血压;3.调节组织液(肿胀或水肿);4.递送氧气和其它营养物质并且移除二氧化碳和其它代谢废物;5.调节体温以及6.减轻久坐不动引起的老化(皱纹、黑眼圈)。除了为整体健康益处保持这些功能之外,微循环还增进皮肤健康。
皮肤微循环是一个复杂而动态的系统,其对于体温调节、皮肤代谢和经皮渗透非常重要。对皮肤的血液供应由微动脉、毛细血管和微静脉的网络所组织成的浅层和深层血管丛提供。皮肤暴露于环境压力源(诸如UV、化学污染物和颗粒物)或生理(非环境的)压力源(心理压力、久坐不动引起的老化以及炎症)中。此类受损的血液流动导致诸如黑眼圈、皱纹、伤口愈合缓慢和水肿的生理效应。
本发明人一直在寻找通过可局部施用或口服摄入的组合物来解决问题(如减少黑眼圈和整体皮肤健康)的方案。发明人希望使用可以大量存在于自然界中的活性物质(或活性物质的组合)来实现这一目标。他们已经发现:包含高于0.025%羟基蒽醌和羟基萘醌的茜草(Rubia Cordifolia)提取物与包含高于0.02%紫铆因的紫矿(Butea monosperma)提取物的组合,当将其局部施用或摄入时,会形成增强的微循环从而产生以上益处。
茜草和紫矿均已被单独用于局部应用但没有同时用于单一组合物中。
US2008206373公开了一种个人护理组合物,其包含至少一种提取物,所述提取物选自油榄仁(Terminalia bellerica)、紫矿、粗糠柴(Mallotus philippinensis)、宽叶榆绿木(Anogeissus latifolia)、藏木香(Innula racemosa)、孟加拉榕(Ficus benghalensis)、夹竹桃(Nerium indicum)、补骨脂(Psoralea corylifolia)、儿茶(Acacia
catechu)、冷杉(Abies pindrow)、雪松(Cedrus deodara)、余甘子(Emblica officinalis)、辣木(Moringa oleifera)、洋甘草(Glycyrrhiza glabra)的提取物以其混合物,和皮肤病学上可接受的载体。
KR20090119340公开了一种用于皮肤增白的化妆品组合物,其含有茜草提取物以确保对抑制黑素生成及酪氨酸酶活化的效果,其已应用于皮肤美白。
印度专利申请1099/MUM/2008公开了含有蒽醌衍生物的茜草植物提取物,所述提取物经评估具有抗微生物活性并且还提供抗炎和抗刺激物特性。
迄今为止,还不知道茜草与紫矿提取物的组合会形成增强的微循环,当局部施用时,其产生减少黑眼圈、头皮健康、减少腋下发黑的益处,且当口服摄入时,表现为心血管疾病的减少。
发明概述
本发明提供了用于增强微/大血液循环的局部、口服或肠胃外组合物,其包含
(i) 0.1至10%的包含高于0.025%羟基蒽醌和羟基萘醌的茜草提取物;
(ii) 0.1至10%的包含高于0.02%紫铆因的紫矿提取物。
优选地,所述组合物包含按重量计0.1至5%的茜草提取物和按重量计0.3至3%的紫矿提取物。
发明详述
通过阅读以下详细描述和所附权利要求,这些和其它的方面、特征以及优点对于本领域普通技术人员将变得显而易见。为了避免疑问,本发明的一个方面的任何特征可被用于本发明的任何其它方面。“包含(comprising)”一词的意思是“包括(including)”但不一定“由…组成”或“由…构成”。换言之,所列举的步骤或选项不必是详尽的。需要注意的是,在以下说明中所给出的实例旨在澄清本发明而并非旨在将本发明局限于那些实例本身。相似地,除非另外指明,否则所有的百分比均为重量/重量百分比。除了在操作和比较实例中,或另行明确指出,在本说明书及权利要求书中表明材料数量或反应条件、材料的物理性质和/或用途的所有数字应当理解为由“约”一词修饰。以“从x到y”的格式表达的数值范围应当理解为包括x和y。当针对特定特征以“从x到y”的格式描述了多个优选的范围时,其被理解为也预期涵盖了不同端值组合的所有范围。
如本文中所用的“局部组合物”意在包括为了更好的皮肤健康益处而应用于哺乳动物(特别是人类)的外表面(例如,皮肤)的组合物。此类组合物一般可以被分为免洗型或洗去型(优选免洗型)并且包括施用于人体的、主要用于增进皮肤健康但也可以用于改善外观、清洁、气味控制或一般美观的任何产品。本发明的组合物可以是液体、乳液、洗液、霜剂、泡沫、凝胶、条皂、棒状物、条状物、衬垫或贴剂的形式。此类局部组合物的非限制性实例包括:免洗型润肤液和润肤霜、止汗剂、除臭剂、唇膏、粉底、睫毛膏、美黑剂或防晒乳液以及洗去型产品如洗发剂、护发素、沐浴露或香皂。如本文中所用的“皮肤”意在包括面部和身体(例如,颈部、胸部、背部、臂部、腋下、手部、腿部、臀部和头皮)的皮肤,并且特别是面部的眼部下方部分。本发明的局部组合物特别可用于对起皱的或更容易起皱的皮肤区域应用(特别是暴露于日光下的身体部分)。
通过食物产品意指用于口服摄入或静脉内注射进入哺乳动物特别是人的产品。通过口服摄入途径而引入人体的食物产品是特别优选的。食物产品也被称为可食用产品,其特别优选用于递送本发明的协同组合,其可以是液体形式诸如汤或饮料(如茶或咖啡)或非液体形式如涂抹食品、调味料、甜品或面包。通过饮料(如茶饮料)递送是特别优选的。可食用产品可以是固体或粉状食品补充剂的形式。
组合物包含茜草和紫矿提取物。水提物是特别优选的。就本发明而言,水提物是已经首先用水提取过的植物来源的提取物。然而,该提取物可以使用其它溶剂进一步分馏。组合物包含0.1至10%的包含高于0.025%羟基蒽醌和羟基萘醌的茜草提取物。羟基蒽醌具有通式结构:
在以上结构中,侧基R1至R8优选地选自以下给出的基团:
R1、R4、R5和R8选自H或OH之一;
R2选自H、OH、CH3或COOH;
R3和R6选自H、OH或CH2OH之一;
R7选自H、CH3或OCH3,并且
R8为H。
更优选地,羟基蒽醌优选以下的一种或多种:
在上表中AQ指的是蒽醌。
存在于用于本发明的组合物中的茜草提取物中的优选的羟基萘醌为:
。
茜草属(茜草科)是一种多年生、草本、纤细、分枝的攀缘植物,有很长的圆柱状的根,曲折,具有红色薄皮,其广泛分布于印度、中国、温带亚洲、非洲和热带澳大利亚。茜草具有很高的商业和医药重要性。在医药上,茜草作为活性物质的重要来源而用于Ayurveda、传统中医体系中的各种处方以及其它现代药物。在本发明中,优选地将植物的茎部用于组合物。
用于本发明的茜草提取物优选地使用两种提取方法之一进行提取以制备两种类型的提取物、所述提取物均基本上为水提物的:(i)水提物或(ii)水提物的酚级分。
用于制备茜草水提物的优选方法:
将茜草的茎部做成粉末并用蒸馏水(1:10的w/v)在80˚C回流约6小时提取,然后用棉布过滤。然后用旋转蒸发仪在45˚C、减压条件下将滤液干燥。得到粘性粉末并将之称为水提物。该提取物通常包含约0.025至10%的羟基蒽醌和羟基萘醌的混合物。
用于制备茜草水提物的酚级分的优选方法:
将以上制备的水提物(10 g)溶于1 L
0.1M NaOH溶液中。然后用1 L 乙酸乙酯(1L)提取该溶液。将乙酸乙酯层从混合物分离。使用1 M HCl中和水层,然后用氯仿提取。收集氯仿层并用盐水溶液洗涤,然后用旋转蒸发仪干燥。所得干燥粉末被称为酚级分或羟基蒽醌和羟基萘醌富集级分。得到约0.76
g 此级分。该提取物通常包含20至约90%的羟基蒽醌和羟基萘醌的混合物。
因此有可能通过上述提取方法来制备包含0.025至90%的羟基蒽醌和羟基萘醌的混合物的茜草提取物。因此,此类提取物优选地包括在本发明的组合物中。茜草提取物以组合物重量的0.1至10%存在。
紫矿通常被称为凤凰木(Flame of forest)并属于豆科(Fabaceae)。在印度,其被冠以各种名字如紫铆(palas)、palash、mutthuga、bijasneha、塔格(dhak)、khakara、chichra、吉纳(Bastard Teak)、孟加拉奇诺(Bengal
Kino)和Nourouc。除了非常干旱的地区,其通常遍布于印度、缅甸和锡兰。其通常生长在开阔的草地上以及散布在混交林中。该植物的几乎所有部分均被长期用于医药应用和用于其它目的。在本发明中,优选地将植物的花的提取物用于本发明的组合物。
将两种类型的紫矿提取物优选的用于本发明的组合物:水提物和经富集的提取物。
用于制备水提物的优选方法:
将100朵干燥紫矿花用800
ml 水在室温、真空(800 mbarr)下提取24小时。将约730 ml 水提物从花分离。将水提物蒸发至干燥以得到约25 g 固体。该提取物通常包含约0.1至4 重量%的紫铆因。
用于制备经富集的提取物的优选方法:
将从以上制备的水提物得到的固体溶于200 ml 水中并通过MCI凝胶填充的柱。用水冲洗MCI基座以移除未吸附的糖类。用乙醇洗脱吸附的级分以得到经富集的级分-1(紫铆因含量为2%)。将经富集的级分-1蒸发至干燥以得到~23 g 固体。
将该固体溶于10% 乙醇水溶液(500 ml),然后用乙醚溶剂提取以得到经富集的级分-2(紫铆因含量为25%)。将该提取物蒸发至干燥以得到约8 g 固体粉末。该提取物通常包含约2至30%重量的紫铆因。
组合物包含0.1至10%的包含高于0.02 %紫铆因的紫矿提取物。紫铆因具有结构:
。
因此有可能通过上述提取方法来制备包含0.02至30%的紫铆因的紫矿提取物。因此,此类提取物优选地包括在本发明的组合物中。紫矿提取物包含高于0.02%的紫铆因,优选在提取物重量的0.1至30%之间。
不希望被理论所束缚,发明人相信两种植物提取物的组合按本发明通过不同的机制工作以提供所需的协同益处。一种机制是通过增加一氧化氮在所需细胞中的生产。一氧化氮(NO)是影响血管紧张度的重要信号分子。在内皮细胞中经eNOS酶提高的一氧化氮生产与多种益处相关。这些益处随着内皮(微内皮/大内皮)细胞的类型而变化。据信,eNOS酶的调节机制在这两种细胞类型中是相同的。当局部施用时,微内皮细胞中更高的NO导致微循环增强、解毒作用增强、伤口愈合增强、炎症后超色素沉着减少。大内皮细胞中更高的NO与心血管疾病的减少、血压降低、动脉粥样硬化和其它大血管并发症的减少相关。本发明人已经发现:当使用具有增强量的多酚(即羟基蒽醌和羟基萘醌)的茜草提取物时,其活化eNOS从而增加内皮细胞中的NO生产。NO生产的增加是经由eNOS磷酸化作用实现的。存在若干种活化该酶的磷酸化位点的机制,例如,它们可能是受体-配体相互作用或超极化。其准确详细的作用机制尚不清楚。值得注意的是,用于体外分析的浓度将会显著低于用在局部或可食用组合物中的水平以得到相似的益处。当用于体外实验(其中细胞与提取物和活性物质直接接触)时,所得到的益处以及提取物的浓度将比考虑了生物有效性和渗透而配制的组合物低约1000倍。
据信,紫铆因的作用是通过不同的机制。紫铆因,紫矿提取物的一种成分,不影响eNOS酶。然而,其在调节II期抗氧化酶、降低精氨酸酶活性和增加自由基清除酶中有作用。已知此机制会抑制活性氧类别(ROS),其将清除产生于细胞内的NO。清除机制是由于NO与自由基(如初生态氧或超氧化物)的反应,其可导致过氧化亚硝酸的形成。据信,过氧化亚硝酸的形成具有负面影响,因为其可通过触发更高的超氧化物而使情况恶化。本发明人相信:通过本发明的方法的协同益处是由于这两种机制的组合 - 一种影响eNOS活性,而另一种则影响根除其它可以使产生的NO淬灭的反应物质而出现的。
本发明的组合物可作为局部组合物施用。此类局部组合物优选地包含化妆品可接受的基底。化妆品可接受的基底可以选自液体、乳液、洗液、霜剂、泡沫、凝胶、条皂、棒状物、面膜、衬垫或贴剂。
可选地,本发明的组合物可以被配制成用于口服的食品组合物。口服组合物优选地被配制成饮料(如汤、茶或咖啡)或非液体形式(如涂抹食品、调味料、甜品、面包)或固体或粉状食品补充剂。
根据本发明的另一个方面,提供了改善皮肤微循环的方法,其包括将本发明的组合物施用于皮肤。
现在,本发明将借助以下非限制性实例进行说明。
实施例
所用材料
DAF FM-DA购自Invitrogen(Eugene, Oregon)。Dolbeco’s
Modified Essential Medium(DMEM)购自Sigma(St. Louis, MO,
USA),胎牛血清(Foetal
bovine serum,FBS)购自Gibco。
茜草和紫矿的样品来源于Bangalore, India的供应商Channabasappa and Co.。样品的地理来源,就茜草而言为印度、尼泊尔和伊朗,而就紫矿而言为印度。
制备培养基:
制备
Dolbeco
’
s Modified Essestial Medium
(
DMEM
)。
该培养基的制备如下:在890 ml 高压消毒蒸馏水中加入1小瓶(13.4克)DMEM(Sigma-Aldrich;目录号 #
D5796 SIGMA),然后加入100 ml FBS(Gibco®;目录号 #
16000-044)和10 ml 青霉素-链霉素溶液(Sigma-Aldrich;目录号 # P4333 SIGMA)。
在CO2培养箱内以37ºC、5% CO2的条件,于含有10% v/v FBS(来自Gibco)的DMEM生长培养基(如上所制备)中生长人类内皮细胞系(EA.hy926,1X105
细胞/10 ml DMEM)(American Type Culture Collection(ATCC);CRL-2922 - ATCC)。
细胞毒性测定:
在开始实验前,有必要确定不同提取物的细胞毒性剂量。不同提取物的细胞毒性通过如下MTT((3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2H-四氮唑溴)测定法进行确定:
所培养的人类内皮细胞系EA.hy926细胞购自American
Type Culture Collection(CRL-2922 - ATCC)并在补充了2 mM L-谷氨酰胺、100 U/ml 青霉素、100 μg/ml 链霉素和10% 体积/体积 FBS(Gibco, Invitrogen)的DMEM(Sigma)中培养。在37oC下、95% 潮湿空气和5% CO2中培养细胞。在达到70-80%汇合度后,通过胰酶消化(0.25% Tryp-EDTA,进行2-3 分钟)对细胞进行继代培养。出于实验目的,将细胞接种到细胞培养板(CLS3524 Sigma)上。
将2x106 人类内皮细胞系(EaHy926)悬浮于10 ml DMEM生长培养基中。将100
µL 该悬浮液放入96孔平板底板(Thermo
Scientific, Nunc;目录号:161093)的每个孔中。因此每个孔具有2x104 个细胞的细胞浓度。然后在CO2培养箱(Thermo Scientific;Forma*
Series II 3110 Water-Jacketed CO2 Incubator)中,于37ºC、5% CO2的条件下让细胞贴壁4小时。4小时后,在DMEM中制备不同浓度(浓度范围:0.1 –100 µg/ml)的提取物,并将其加入到细胞中并进一步培养24小时。为了制备以上浓度范围内的提取物,首先将提取物溶于DMSO(二甲基亚砜,Sigma-Aldrich;目录号D5879)中以达到50 mg/ml的原液浓度。将提取物的DMSO原液进一步重悬浮于DMEM中(浓度范围:0.1–100 µg/ml)。24小时后,用磷酸盐缓冲盐水(PBS)缓冲液洗涤细胞并在CO2培养箱中用100
µL MTT试剂(Sigma-Aldrich;目录号M5655 SIGMA)处理4小时。通过将粉末状MTT溶于PBS中来制备MTT试剂(浓度:5 mg/mL)。4小时后,移除上清液并收集细胞内部所形成的MTT甲晶体,并将其溶于DMSO中并且使用Biorad multiplate reader(Bio-Rad;型号680)在540 nm波长测量吸光度。
观察到当经过24小时温育,最大非细胞毒性浓度对于茜草提取物为20 μg/ml,而对于紫矿提取物为50 μg/ml。因此对于后续实验,使用的茜草提取物的非细胞毒性浓度为20 μg/ml或更低,而紫矿提取物为50 μg/mL或更低。还发现:在长达4小时中,茜草提取物的最大非细胞毒性浓度为50 μg/ml。
制备
EA.Hy926
细胞用于一氧化氮测定
所培养的人类内皮细胞系EA.hy926细胞购自American
Type Culture Collection(CRL-2922 - ATCC)并在补充了2 mM L-谷氨酰胺、100 U/ml 青霉素、100 μg/ml 链霉素和10% 体积/体积 FBS(Gibco, Invitrogen)的DMEM(Sigma)中培养。 在37oC下、95% 潮湿空气和5% CO2中培养细胞。在达到70-80%汇合度后,通过胰酶消化(0.25% Tryp-EDTA,进行2-3 分钟)对细胞进行继代培养。 出于实验目的,将细胞接种到24孔组织培养板(Sigma-Aldrich;Z688789
SIGMA)上。在贴壁后,在进行任何实验之前让细胞在无血清低葡萄糖DMEM中经受12-14小时饥饿,以使细胞保持在静止状态并降低NO生产的基础水平。
使用流式细胞术测量细胞内一氧化氮
实验流程:
1. 将EAHy926(5x105)细胞接种到24孔组织培养板中并让其贴壁12-16小时。
2. 贴壁后,在进行任何实验之前让细胞在无血清低葡萄糖DMEM中经受12-14小时饥饿。
3. 贴壁后,细胞经过或不经过包含活性物质A(紫铆因富集的提取物)的提取物的处理并在不含FBS的DMEM中温育24小时。
4. 24小时后,向24孔组织培养板中的细胞加载DAF
FM-DA(1 μM)30 分钟,使用无血清培养基洗涤两次。
5. 随后,在37oC下、95% 潮湿空气和5% CO2中,用不同浓度的活性物质B(茜草提取物)刺激细胞45分钟。
6. 然后将经刺激的细胞用无血清培养基洗涤两次。
7. 用0.25% 胰蛋白酶-EDTA对细胞进行胰酶消化,并用2% PFA固定15 分钟。
8. 采用流式细胞术采集细胞悬浮液。以10,000个细胞的群作门控并用FACS Calibur(BD;SanDiego)测量其相对荧光强度。在经处理和未处理的细胞之间比较细胞的荧光强度(反映了NO生产)。
如下表1中,将结果制成表格。
在上表中,酚类茜草提取物是指具有约85%羟基蒽醌和羟基萘醌的提取物。
经富集的紫矿提取物是指具有约25%紫铆因的提取物。基于所取的量,组织培养板的每个孔中的实际浓度也在上表中标明。
上表中的数据表明:紫矿提取物和茜草提取物的组合在指示增强的微循环的体外测定中提供了协同活性。
茜草和紫矿提取物浓度的影响:
使用以上提及的程序测试了各种浓度的茜草和紫矿提取物的影响。由于在长达4小时中,茜草提取物的最大非细胞毒性浓度为50 μg/ml,因此在这些实验中所选用的茜草和紫矿的浓度均为最高达50 μg/ml。数据在表2中提供。
表2中所提供的结果显示:与使用单独的提取物所得到的益处相比,当使用紫矿和茜草提取物的组合时,得到了增强的或协同的益处。当浓度为0.0009%的蒽醌(对应于10 μg/ml的茜草酚类提取物)与0.0003%的紫铆因(对应于10 μg/ml的紫矿提取物)组合时,益处是明显的。所获得的益处和表中提及的浓度是基于体外实验,其中细胞与提取物和活性物质直接接触。受限于生物有效性和渗透,要达到观测到的益处所需的浓度在所配制的组合物中将是大约1000倍。
当任一提取物处于低浓度(即紫矿或茜草处于1 μg/ml)时,未获得协同益处。
Claims (5)
1.一种局部或口服或肠胃外组合物,其用于增强微/大血液循环,其包含:
(i) 0.1至10%的包含高于0.025%羟基蒽醌和羟基萘醌的茜草提取物;
(ii) 0.1至10%的包含高于0.02%紫铆因的紫矿提取物。
2.根据权利要求1的组合物,其中所述组合物包含按重量计0.1至5%的茜草提取物和按重量计0.3至3%的紫矿提取物。
3.根据权利要求1的局部组合物,其另外地包含化妆品可接受的基底,所述基底选自乳液、洗液、霜剂、泡沫、凝胶、条皂、棒状物、面膜、衬垫或贴剂。
4.根据权利要求1的口服组合物,其中所述组合物为饮料如汤、茶或咖啡;或为非液体形式如涂抹食品、调味品、甜品、面包;或为固体的或粉状的食品补充剂。
5.所述组合物在改善皮肤血液微循环上的用途,其包括将根据权利要求3所述的组合物施用于皮肤或摄入根据权利要求4所述的组合物。
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CN101360478A (zh) * | 2006-01-19 | 2009-02-04 | 株式会社太平洋 | 含有红松提取物作为活性成分的化妆品组合物 |
CN102125585A (zh) * | 2010-01-12 | 2011-07-20 | 株式会社布伦克蕾喜 | 活性氧清除剂、自由基清除剂以及氧化性细胞障碍抑制剂 |
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WO2015018631A1 (en) | 2015-02-12 |
EP3041586B1 (en) | 2016-11-30 |
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