CN109394557A - 包含芥子酸的皮肤皱纹改善或弹性增强的组合物 - Google Patents
包含芥子酸的皮肤皱纹改善或弹性增强的组合物 Download PDFInfo
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- CN109394557A CN109394557A CN201810930380.7A CN201810930380A CN109394557A CN 109394557 A CN109394557 A CN 109394557A CN 201810930380 A CN201810930380 A CN 201810930380A CN 109394557 A CN109394557 A CN 109394557A
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- skin
- enhancing
- wrinkle
- sinapic acid
- collagen
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Abstract
本发明涉及包含芥子酸(Sinapinic acid)或其盐作为有效成分的用于改善皮肤皱纹、增强弹力、增强皮肤保湿、去除角质、抑制红斑或改善皮肤光老化的组合物以及保健食品组合物。更详细地,本发明的芥子酸具有增加前胶原蛋白分泌量、促进胶原蛋白的生物合成性、减少MMPs基因的表达、增加弹力蛋白(elastin)基因的表达、抑制红斑或抑制皮肤表皮层的增厚的效果,能够有效改善皮肤皱纹、增强弹力、增强保湿、去除角质、抑制红斑或改善皮肤光老化。
Description
技术领域
本发明涉及包含芥子酸(Sinapinic acid)或其化妆品学上允许的盐作为有效成分的用于改善皮肤皱纹或增强弹力的化妆品原料组合物。
背景技术
最近,随着医疗技术的发展带来的平均寿命的延长及人们对生活质量的提高和美好健康生命的追求的增加,对皮肤美容以及健康的关心日益增长。迎合人们要维持健康皮肤的目的,开发出了各种美容功能性化妆品,尤其预防、缓解和改善皮肤皱纹的研究非常活跃。同时,人们的意识也发生了变化,认为近来的化妆品成分在到达皮肤真皮供给营养成分的方面受限,也可通过摄取食品来为皮肤供给营养成分或功能成分而达到皮肤美容效果,从而挖掘内在美(inner beauty)食品素材的研究也正在活跃地进行(Chung,S.et al.,γ-Linolenic acid in borage oil reverses epidermal hyperproliferation in guineapigs.J Nutr,132,3090-3097,2002;Tanno,O.et al.,Nicotinamide increasesbiosynthesis of ceramides as well as other stratum corneum lipids to improvethe epidermal permeability barrier.Br J Derm,143,524-531,2000)。
皮肤大致由表皮(epidermis)、真皮(dermis),皮下脂肪(hypodermis) 这三层构成。表皮,尤其是表皮的最外层的角质层起到皮肤屏障作用,能够抑制水分或电解质通过皮肤流失,另一方面,真皮层起到通过合成胶原蛋白和弹力蛋白来维持皮肤的弹力并支撑构造的作用。即,胶原蛋白和弹力蛋白是由成纤维细胞生成的主要蛋白质,关系到皮肤的机械性坚固性、组织结合力及弹性等(Moskowit,R.W.et al,Role of collagen hydroiysatein bone and joint disease,Semin Arthritis Rheu,30:87,2000)。胶原蛋白根据形态和结构特征构成各种同工型(isoform),人体组织中共存在28种胶原蛋白同种型 (isotype),其中作为皮肤组织中存在的胶原蛋白,已知的是1、3、4、6、7、 13、14、17型胶原蛋白等。1型和3型胶原蛋白构成细胞间质构成成分,7 型胶原蛋白成为真皮与表皮交界处(dermal andepidermal junction)的主要构成物质(Pei,M.et al.,Expression of collagen type I,II and III in loose body of osteoarthitis,J.Orthop.Sci.,5:288,2000)。
皮肤结合组织中,细胞外基质蛋白质中存在最多的是I型胶原蛋白,除此之外还存在弹力蛋白、纤连蛋白(fibronectin)、整联蛋白(integrin)、原纤维蛋白(fibrillin)、聚糖等蛋白质。新合成的前胶原蛋白经过酶促反应分泌到皮肤细胞的细胞外空间之后形成三螺旋结构(triple helix configuration) 的微纤维(microfibril),微纤维与富含亮氨酸的小蛋白聚糖(leucine-rich small proteoglycans)结合而形成原纤维(fibril)。最终,如此合成的原纤维聚集而形成用于提供皮肤的结合力和弹性的胶原纤维(Bateman,J.F.etal.,Collagen superfamily.Extracellular matrix,Harwood,New York,2,22-26,1996)。
据悉,皮肤老化是因为胶原蛋白的含量减少,胶原蛋白作为在皮肤真皮组织的胶原质中占大部分的蛋白质,为皮肤带来张力和强度,因而胶原蛋白的减少与皮肤老化和皱纹形成存在紧密联系。皮肤老化大致分为生理学老化引起的内因性老化以及持续暴露在紫外线(ultraviolet radiation,UV)引起的光老化。反复暴露在紫外线会增加胶原酶,引发胶原纤维的变性和破坏,而降低皮肤弹力、促进皱纹生成。即,持续暴露在紫外线的皮肤组织中会增加活性氧簇(ROS)的产生,后者会通过表皮生长因子受体(epidermal growthfactor receptor,(EGF-R)、肿瘤坏死因子受体(tumor necrosis factor –receptor,TNF-R)等介导的信号传递体系而促进促炎性细胞因子的生成 (Sachsenmaier,C.et al.,Involvement of growth factor receptors in the mammalian UVC response.Cell,78:963-972,1994;Dy,L.C.et al., Augmentation of ultraviolet B radiation-inducedtumor necrosis factor production by the epidermal platelet-activating factorreceptor.J.Biol.Chem.: 274,26917-2692,1999),这些受体的活性化最终会构成使得用于介导包含胞外信号调节激酶(mitogen-activated protein kinase,MAPK)在内的下游(downstream)的信号传递的蛋白质活性化的连续性磷酸化,结果会使活化剂蛋白-1(activator protein-1,AP-1)和核因子κB(nuclear factorκB,NF-κB) 等转移因子活性化而引发炎症反应,并促进基质金属蛋白酶(matrix metalloproteinase,MMPs)等胶原酶(collagenase)的活性,从而降低皮肤弹力并促进皱纹生成(Kang,S.et al.,Inflammationand extracellular matrix degradation mediated by activated transcriptionfactors nuclear factor-kB and activator protein-l in Inflammatory acnelesions in vivo.Am J.Pathol.,166: 1691-1699,2005)。AP-1调节与细胞的生长和分化有关的大量基因的表达,并强力调节部分MMPs的表达,由AP-1调节其表达的MMP中MMP-1已知为胶原酶1(collagenase 1),将1型和3型胶原蛋白作为基质。
发明内容
本发明的目的是提供一种包含芥子酸(Sinapinic acid)或其化妆品学上允许的盐作为有效成分的用于改善皮肤皱纹或增强弹力的化妆品原料组合物等。
但是,本发明要解决的技术问题不限于以上提及的问题,本领域普通技术人员可从以下记载明确未提及的其它问题。
本发明提供一种有芥子酸(Sinapinic acid)或其化妆品学上允许的盐作为有效成分的用于改善皮肤皱纹或增强弹力的化妆品原料组合物。
上述芥子酸(Sinapinic acid)能够起到如下作用:ⅰ)增加前胶原蛋白(procollagen)的分泌量,ⅱ)减少基质金属蛋白酶(matrixmetalloproteinase, MMP)基因的表达,ⅲ)增加Ⅰ型胶原α1(collagen type Ⅰ alpha 1,Col Ⅰ α1)、Ⅰ型胶原α2或Ⅲ型胶原α1(Col3α1)的表达,或者ⅳ)增加弹力蛋白(elastin)基因的表达。
上述化妆品原料组合物还可包含由维生素C、视黄酸、转化生长因子(transforming growth factor,TGF)、动物胎盘来源的蛋白质、桦木酸以及小球藻提取物构成的组中的一种以上。
本发明提供一种包含芥子酸(Sinapinic acid)或其化妆品学上允许的盐作为有效成分的用于增强皮肤保湿、去除角质、抑制红斑或改善皮肤光老化的化妆品原料组合物。
上述化妆品原料组合物抑制皮肤表皮层的增厚。
上述化妆品原料组合物抑制真皮与表皮交界处的胶原纤维(collagen fiber)网受损。
本发明提供一种包含芥子酸(Sinapinic acid)或其食品学上允许的盐作为有效成分的用于改善皮肤皱纹或增强弹性的保健食品组合物。
本发明提供一种包含芥子酸(Sinapinic acid)或其食品学上允许的盐作为有效成分的用于增强皮肤保湿、去除角质、抑制红斑或改善皮肤光老化的保健食品组合物。
本发明涉及包含芥子酸(Sinapinic acid)或其盐作为有效成分的用于改善皮肤皱纹、增强弹力、增强皮肤保湿、去除角质、抑制红斑或改善皮肤光老化的组合物,上述芥子酸促进皮肤维芽细胞中前胶原蛋白的分泌,增加活性化的胶原蛋白的量,增加皮肤真皮层中胶原纤维(collagen fiber)的量,抑制催化胶原蛋白的分解的MMPs基因的表达。能够用于改善皮肤皱纹或增加弹力。
并且,上述芥子酸增加皮肤组织的含水量,降低水分蒸发量,具有皮肤保湿改善效果,能够抑制红斑,减少变得增厚的皮肤表皮层的厚度,能够去除角质或防止紫外线等引导的皮肤光老化。
附图说明
图1示出人皮肤成纤维细胞中前胶原蛋白的分泌量的测定结果。
图2a至图2d示出摄取根据实施例2、比较例3以及比较例4制备的实验食物的小鼠的体重增加量及食物摄取量的测定结果。
图3a至图3d示出摄取根据实施例2、比较例3以及比较例4制备的实验食物的小鼠的皮肤组织的水分量、弹性以及红斑指数的测定结果。
图4是示出摄取根据实施例2、比较例3以及比较例4制备的实验食物小鼠的背皮肤组织的照片。
图5a至图5b示出摄取根据实施例2、比较例3以及比较例4制备的实验食物的小鼠等的背皮肤组织的皱纹形成程度的测定结果。
图6示出摄取根据实施例2、比较例3以及比较例4制备的实验食物的小鼠的皮肤厚度的测定结果。
图7示出摄取根据实施例2、比较例3以及比较例4制备的实验食物的小鼠的胶原纤维(collagen fiber)量及形态变化。
图8示出经本发明的芥子酸(Sinapinic acid)处理的无毛小鼠的背组织中胶原蛋白及MMPs基因的表达变化。
图9示出经本发明的芥子酸(Sinapinic acid)处理的无毛小鼠背组织中弹力蛋白(Elastin)基因的表达变化。
具体实施方式
本发明人经确认,芥子酸促进皮肤成纤维细胞中前胶原蛋白的分泌,增加皮肤真皮层中胶原纤维的量,抑制MMPs基因的表达,促进皮肤组织中含水量以及抑制水分蒸发,从而具有改善皮肤皱纹、增强弹力、增强皮肤保湿的效果,减少红斑指数以及减少变得增厚的皮肤表皮层的厚度,从而具有抑制红斑、去除角质以及改善皮肤光老化的效果,由此完成了本发明。
本说明书中,“改善皮肤皱纹”是指维持或强化与皱纹及弹力相关的能力。作为皮肤真皮层的胶原纤维的胶原蛋白(collagen)和作为弹力纤维的弹力蛋白(elastin)是起到这种作用的主要蛋白质,主管皮肤弹力,胶原蛋白的生物合成受到皮肤的内外影响。具体地,皮肤细胞如果由于自然老化而致使细胞活性减少,则引发胶原纤维的减少,或者作为外因被紫外线过量照射或因压力等生成的活性氧与蛋白质的硫醇基(thiol:-SH)反应而阻碍酶活性或增加胶原蛋白、弹力蛋白等的分解酶的表达来增加皮肤皱纹并减少弹力而造成皮肤老化。
本说明书中,“皮肤光老化”是指由于紫外线暴露而致使皮肤老化的生理现象,如果长期暴露在太阳(紫外线),脸部、颈部皱纹会增加,皮肤变更干燥,皮肤表面变得粗糙,或者引发雀斑等色素沉着。
用于改善皮肤皱纹或增强弹力的化妆品原料组合物
本发明提供一种包含芥子酸(Sinapinic acid)或其化妆品学上允许的盐作为有效成分的用于改善皮肤皱纹或增强弹力的化妆品原料组合物。
具体地,上述芥子酸由以下化学式1表示,结构式为C19H22O2,分子量为294.4g/mol,IUPAC(国际纯粹与应用化学联合会)名称为3,5-二甲氧基-4-羟基苯丙烯酸[3-(4-hydroxy-3,5-dimethoxyphenyl)prop-2-enoic acid]。
并且,在具有与上述芥子酸同等功效的范围内,可包含芥子酸水合物、芥子酸衍生物等,还可包含其溶剂化合物或立体异构体。
化学式1
上述芥子酸的取得方法没有特别限制,可从含有上述芥子酸的食物分离或利用公知制法进行化学合成或从市场中购买使用。
上述芥子酸能够起到如下作用:ⅰ)增加前胶原蛋白(procollagen)的分泌量,ⅱ)减少基质金属蛋白酶(matrixmetalloproteinase,MMP)基因的表达,ⅲ)增加Ⅰ型胶原α1(collagen type Ⅰ alpha 1,Col Ⅰ α1)、Ⅰ型胶原α2或Ⅲ型胶原α1(Col3α1)的表达,或者ⅳ)增加弹力蛋白(elastin) 基因的表达。
具体地,作为构成皮肤的主要蛋白质的胶原蛋白在存在于皮肤真皮中的成纤维细胞合成为前胶原蛋白的形态之后以细胞外基质分泌,以细胞外基质分泌的前胶原蛋白由于存在于细胞表面的前胶原肽酶(procollagen peptidase)而致使C-末端分解并转换为活性型胶原蛋白,因而测定C肽 (C-peptide)含量即可测定出活性化的胶原蛋白的含量。即,本发明的芥子酸能够促进皮肤成纤维细胞中前胶原蛋白的合成,从而增加活性化的胶原蛋白的量。
并且,催化胶原蛋白分解的基质金属蛋白酶(matrixmetalloproteinase, MMP)在哺乳类中存在23种类型,其中作为由于紫外线而增加的MMP类型已知的是1型、3型及9型,这三种类型的MMPs是分解1型、3型胶原蛋白的酶。具体地,MMP-1起到切断胶原纤维的中间的作用,MMP-3、MMP-9 起到将上述切断的胶原纤维切断为三等分的作用。即,本发明的芥子酸能够明显减少MMPs基因的表达,从而抑制胶原纤维的分解,增加皮肤组织中胶原蛋白蛋白质的合成,抑制皱纹的形成。
皮肤的真皮由作为胶原纤维的胶原蛋白和作为弹力纤维的弹力蛋白构成。上述胶原纤维形成皮肤张力并构成结构性统合,弹力蛋白纤维关系到皮肤弹力,能够增强皮肤弹力。具体地,本发明的芥子酸促进弹力蛋白表达,从而达到增强皮肤弹力的效果。
进而,本发明的化妆品原料组合物还可包含由维生素C、视黄酸、转化生长因子(transforming growth factor,TGF)、动物胎盘来源的蛋白质、桦木酸以及小球藻提取物构成的组中的一种以上,优选的是维生素C,但不限于此。具体地,根据本发明的一实施例确认到,将芥子酸和维生素C一同处理到皮肤成纤维细胞时,与单独处理上述芥子酸或维生素C的情况相比,前胶原蛋白的分泌量增加10p%至11p%,根据Colby公式证明了显著上升效果。即,芥子酸与维生素C一同使用的情况比单独使用芥子酸的情况,皮肤成纤维细胞中前胶原蛋白的分泌增加,能够有效增加活性化的胶原蛋白的量。
上述芥子酸用量优选为在0.1μM至1000μM浓度范围内,但不限于此。此时,如果芥子酸小于上述浓度范围,则前胶原蛋白的合成以及催化胶原蛋白分解的MMPs基因表达抑制程度降低,因而难以发挥改善皮肤皱纹或增加弹力的效果,如果大于上述浓度范围,则有可能存在包括细胞毒性的毒性的忧虑。
如上所述,本发明的芥子酸能够促进前胶原蛋白分泌,从而增加活性化的胶原蛋白的量,减少催化胶原蛋白分解的MMPs基因的表达,增加弹力蛋白基因的表达,预防或缓解皮肤皱纹,并增强皮肤弹力。
本发明的化妆品原料组合物包含芥子酸作为有效成分,同时可添加皮肤学上允许的赋形剂来制备成基础化妆器组合物(化妆水、面霜、精华液、洗面奶以及卸妆水等卸妆液、面膜、润肤油)、彩妆组合物(粉底、口红、睫毛膏、基底液)、头发用产品组合物(洗头膏、护发素、焗油膏、发胶)或香皂等形态。
上述赋形剂不受限制,例如可包含皮肤软化剂、皮肤渗透增强剂、着色剂、芳香剂、乳化剂、浓化剂及溶剂。并且,还可包含香料、色素、杀菌剂、抗氧化剂、防腐剂以及保湿剂等,出于改善物性的目的,还可包含粘增剂、无机盐类、合成高分子物质等。例如,作为本发明的化妆品原料组合物制备卸妆液或香皂的情况下可以在通常的卸妆液和基础油中添加上述芥子酸来容易制得。制备面霜的情况下,可以一般的水包油型(O/W)乳状液添加芥子酸或其盐制得。其中还可添加香料、螯合物、色素、抗氧化剂、防腐剂等用于改善物性的蛋白质、矿物质、维生素等合成或天然素材。本发明的化妆品原料组合物中所包含的芥子酸的含量不限于此,优选为相对于组合物总量包含0.001至10重量百分比,更优选为0.01至5重量百分比。如果上述含量小于0.001重量百分比,无法期待所要达到的抗老化或改善皱纹的效果,如果大于10重量百分比,则存在安全问题或难以制备剂型。
用于改善皮肤皱纹或增强弹力的保健食品组合物
本发明提供一种包含芥子酸(Sinapinic acid)或其食品学上允许的盐作为有效成分的用于改善皮肤皱纹或增强弹性的保健食品组合物。上述芥子酸的具体内容在上面已经表述过。
本发明的用于改善皮肤皱纹或增强弹性的保健食品组合物中,将上述芥子酸用作保健食品的添加剂时,可以直接添加或与其他食品或食品成分一同使用,可根据通常的方法适当使用。有效成分的混合量可根据预防、保健或治疗等使用目的适当决定。
保健食品的剂型不仅可以是散剂、颗粒剂、丸剂、锭剂、胶囊剂等形态,还可以是一般食品或饮料形态。
上述食品的种类不受特别限制,可添加上述物质的食品的例子有肉类、香肠、面包、巧克力、糖类、膨化食品类、饼干类、披萨、方便面、其他面类、口香糖类、包括冰淇淋类的各种乳制品、各种汤、饮料、茶、饮剂、酒精饮料及复合维生素等,可包含通常意义上的各种食品。
通常,在制造食品或饮料时,相对于100重量份的原料,可添加15重量份以下的上述芥子酸,优选的是添加10重量份以下的量。但是,如果是出于健康或卫生的目的或出于保健的目的而需要长期摄取的情况下,可将上述量调节到上述范围以下,并且,本发明利用来自天然物的分馏物,因而不存在安全问题,也可以使用上述范围以上的量。
本发明的保健食品中饮料可像普通饮料一样添加各种香味剂或天然碳水化物等添加成分。上述的天然碳水化物可以是葡萄糖、果糖等单糖,麦芽糖、蔗糖等双糖,糊精、环式糊精等多糖,木糖醇、山梨糖醇、赤藻糖醇等糖醇。作为甜味剂,可使用索马甜、甜叶菊提取物等天然甜味剂,或糖精、阿斯巴甜等合成甜味剂等。上述天然碳水化物的比率可以是每100mL本发明的饮料约为0.01~0.04g,优选为0.02~0.03g。
除上面提到的以外,本发明的用于改善皮肤皱纹或增强弹力的保健食品还可含有各种营养剂、维生素、电解质、调味剂、着色剂、果胶酸及其盐、藻酸钠及其盐、有机酸、保护性胶体增粘剂、pH调节剂、稳定化剂、防腐剂、甘油、酒精、碳酸饮料中使用的碳酸化剂。除此之外,本发明的用于改善睡眠的组合物还可含有用于制造天然果汁、果汁饮料及蔬菜饮料的果肉。这些成分可单独使用或混合使用。这些添加剂的比率不受限制,一般相对于 100重量份本发明的保健食品在0.01~0.1重量份范围内选择。
增强皮肤保湿、去除角质、抑制红斑或改善皮肤光老化的化妆品原料组合物
本发明提供一种包含芥子酸(Sinapinic acid)或其化妆品学上允许的盐作为有效成分增强皮肤保湿、去除角质、抑制红斑或改善皮肤光老化的化妆品原料组合物。上述芥子酸的具体内容在上面已经表述过。
具体地,上述化妆品原料组合物能够增加皮肤组织的含水量,减少水分蒸发量,从而增强皮肤保湿,减少红斑指数来抑制红斑。
并且,上述化妆品原料组合物能够抑制皮肤表皮层的增厚,抑制皮肤真皮与表皮交界处的胶原纤维(collagen fiber)网受损,从而改善皮肤光老化。具体地,如果由于紫外线等造成光老化,为了保护皮肤真皮层而增加形成角质层使得皮肤变厚。由此,紫外线照射使得皮肤表皮层的厚度变厚意味着光老化引起的皮肤损伤严重。根据本发明的一实施例,确认照射了紫外线的小鼠的皮肤厚度变化的结果,紫外线照射致使皮肤厚度明显增加,且上述照射了紫外线的小鼠摄取包含芥子酸的实验食物后,皮肤厚度明显减少。即,本发明的芥子酸能够减少因紫外线照射而变得增厚的表皮层的厚度,从而改善皮肤光老化。
优选地,上述芥子酸的用量在0.1μM至1000μM的浓度范围内,但不限于此。此时,如果芥子酸小于上述浓度范围,则抑制皮肤组织的含水量的增加,增加水分蒸发量,不能有效抑制皮肤表皮层的增厚,因而存在难以发挥皮肤保湿、去除角质、抑制红斑或改善皮肤光老化的效果。而如果大于上述浓度范围,则有可能存在包含细胞毒性的毒性。
如上所述,本发明的芥子酸能够有效维持皮肤组织内水分,减少红斑指数,防止皮肤表皮层的厚度变得增厚,从而增强皮肤保湿,促进去除角质,抑制红斑,并改善皮肤光老化。
增强皮肤保湿、去除角质、抑制红斑或改善皮肤光老化的保健食品组合物
本发明提供一种包含芥子酸(Sinapinic acid)或其食品学上允许的盐作为有效成分的用于增强皮肤保湿、去除角质、抑制红斑或改善皮肤光老化的保健食品组合物。上述芥子酸的具体内容在上面已经表述过。
以下,为了有助于理解本发明,而提出优选实施例。然而,以下实施例只是为了更容易理解本发明而提供的,并不能由此限定本发明的内容。
准备例
准备例1
从ATCC(美国菌种保藏中心,美国美弗吉尼亚洲马纳萨斯)购买人皮肤成纤维细胞(normal human dermal fibroblasts,neonatal foreskin),并利用成纤维细胞生长培养基(fibroblast growth medium)(德国海德堡的Promo Cell公司)在37℃、5%CO2孵化器中对所购细胞进行培养来用于实验。
准备例2
从Oriental Bio公司(韩国京畿道)购入5周龄雄性白化无毛活鼠 (Skh-1),喂食固体饲料经过1周适应期。实验动物分为4个组,每组分配5只用于实验。饲养期间自由摄取饲料和水,温度维持22±1℃,湿度维持60±5%,每天将光周期和暗周期调节为12小时。
实施例
实施例1.I型前胶原蛋白C肽(Procollagen Type IC-peptide)(PIP)浓度测定
将准备例1的人皮肤成纤维细胞在12孔板(well-plate)中按照1.0×106细胞/孔(cells/well)进行分株,按100μM的浓度添加芥子酸(SA),在 CO2培养基中培养24小时。去除各孔(well)的培养基后利用PBS(磷酸缓冲盐溶液)洗涤1次,再加入1ml的PBS之后按照20mJ/cm2条件照射紫外线B(ultraviolet B,UVB)。将各孔(well)的PBS再更换为培养基培养 24小时之后,利用I型前胶原蛋白C肽EIA试剂盒(日本宝生物工程株式会社(TakaraBio))来测定分泌到培养基的前胶原蛋白的量。将胶原蛋白测定试剂盒中包含的标准溶液按照各浓度进行稀释后,在450nm条件下测定吸光度来制成标准浓度曲线,算出胶原蛋白生成量。
比较例1
除了利用100μM的维生素C进行处理之外,按照与上述实施例1相同的方法进行。
比较例2
除了利用100μM的芥子酸及100μM的维生素C进行处理之外,按照与上述实施例1相同的方法进行。
其结果,如图1所示,照射紫外线的对照细胞(+UVB)的情况相比正常细胞(-UVB),前胶原蛋白分泌量显著减少,实施例1相比只照射紫外线的对照细胞(+UVB),胶原蛋白量明显增加,增加了36%。并且,比较例2相比照射紫外线的对照细胞(+UVB),胶原蛋白量明显增加,增加了 47%,该数值比比较例1或实施例1中观察到的胶原蛋白还高。即,本发明的芥子酸具有与已知为具有稳定并促进胶原蛋白的维生素C相似的效果。并且,上述芥子酸与维生素C一同使用时,随着促进前胶原蛋白的分泌,能够增加活性化的胶原蛋白的量。
实施例2.摄取添加了芥子酸的锭剂食物
给准备例2的小鼠供给在遵照AIN-93啮齿动物饮食(rodent diet)构成 (Reeves,PG et al.,J Nutr,123:1939-1951,1993)调配的锭剂食物中添加 0.2%芥子酸(西格玛奥德里奇)制得的食物,供给10周,且于每天上午10-11 点(韩国时间)之间与水一同供给,上述AIN-93啮齿动物饮食的组成如下表1。
实验饲养期间,在无毛活鼠的背部照射紫外线B(ultraviolet B,UVB),每周照射3次,至于紫外线照射量,第一周照射100mJ/cm2,第二周照射 200mJ/cm2,第三周到第十周照射300mJ/cm2。
实验饲养最后一天,在使实验动物空腹6小时后麻醉的状态下,采取血液和背皮肤组织。血液试样利用注射器从腹部下大静脉中采取,并放入涂敷了乙二胺四乙酸(EDTA)的试管(tube)中,在2000×g条件下进行15分钟的离心分离之后分离血浆,在-70℃温度下冷冻保管至进行分析为止。背皮肤组织的一部分立即在70℃温度下冷冻保管后用于分子生物学检查,一部分利用10%福尔马林溶液进行固定,用于免疫组织化学染色。
比较例3
3-1)除了供给遵照AIN-93啮齿动物饮食构成调配的锭剂食物之外,按照与上述实施例2相同的方法进行。
3-2)除了未照射紫外线B(ultraviolet B,UVB)之外,按照与上述比较例3-1)相同的方法进行。
比较例4
除了供给在AIN-93锭剂食物中添加0.2%维生素C(西格玛奥德里奇) 制得的食物之外,按照与上述实施例2相同的方法进行。
表1
实验例1.无毛活鼠的体重及食物摄取量变化
对上述实施例2、比较例3以及比较例4的小鼠,测定体重及食物摄取量。其结果,如图2a至图2d所示,芥子酸及维生素C的摄取对上述小鼠的体重及食物摄取量未造成明显影响。
实验例2.测定皮肤保湿、弹性及红斑指数
针对上述实施例2、比较例3以及比较例4的小鼠,利用皮肤水分测量仪(Corneometer)、皮肤水分流失测量仪(Tewameter)、皮肤弹性测量仪 (Cutometer)、皮肤黑色素和血红素测量仪(Mexameter,CK Electronics GmbH)在实验结束日测定一次皮肤含水量、水分蒸发量、弹性及红斑指数。测定时,记录轻压实验动物等的特定部分显示的数值。
其结果,如图3a至图3d所示,照射10周紫外线的比较例3-1)相比未照射紫外线的比较例3-2),皮肤组织的含水量及弹性明显减少,另一方面水分蒸发量及红斑指数明显增加。并且,实施例2的情况下,尽管照射了相同强度的UV,但相比比较例3-1),含水量及弹性明显增加,分别增加了61%及86%,而水分蒸发量及红斑指数明显减少,分别减少了58%及23%。
实验例3.测定背皮肤组织的皱纹
针对照射10周紫外线的实施例2、比较例3-1及比较例4的小鼠的皮肤,利用硅橡胶制作等效模型之后测定皱纹形成程度。在上述小鼠的背部附着具有直径为1cm的圆形孔的盘(disk),混合等效模型制作用试药,薄薄地展开涂抹到无毛活鼠的背部,完全干燥后,小心地取出盘来制作等效模型。等效模型的制作在温度为20~23℃且湿度为45~50%的恒温恒湿状态下实施,并使用了用于制作等效模型的硅橡胶印模材料(韩国首尔的Epigem公司)。至于制成的等效模型的分析,利用计算机影像分析仪(Visioline VL650,德国CKelectronic GmbH公司)分析出皱纹的总面积(total wrinkle area)、最大皱纹深度(maxwrinkle depth)、皱纹平均深度(mean depth)及平均长度 (mean length)等4项皱纹指标项目。
其结果,如图4所示,比较例3-1)与未照射紫外线的比较例3-2)情况相比形成了粗且深的皱纹的同时形成了小细纹,实施例2虽然照射了相同强度的UV,但相比上述比较例3-1),深皱纹几乎消失,皱纹的粗细和深度均得到了显著改善。并且,如图5b所示,根据利用计算机影像分析仪在等效模型中将皱纹形成程度数值化的结果也可确认到,实施例2与比较例3-1) 的情况相比,皱纹总面积、皱纹平均深度、皱纹平均长度及皱纹最大深度均明显减少,分别减少了80%、38%、69%及64%,这种芥子酸的皱纹改善功效比在维生素C中观察到的皱纹改善功效相似或更好。即,摄取芥子酸能够显著抑制紫外线引起的皱纹生成。
实验例4.皮肤厚度变化
(1)测定背皮肤厚度变化
针对上述实施例2、比较例3以及比较例4的小鼠臀部的皮肤厚度,利用数字微卡尺(digital micro caliper)(加拿大安大略的马拉松手表有限公司(Marathon WatchCompany Ltd))进行测定。测定时使用的卡尺能够测定至0.01mm,且具有能够对厚度施加一定力的调节功能,因而能够在施加同行力的状态下测定皮肤厚度。
其结果,如图6的(A)部分所示,比较例3-1)相比未照射紫外线的比较例3-2)的情况,皮肤厚度明显增加,实施例2与比较例3-1)相比,皮肤厚度明显减少,减少了71%。
(2)皮肤组织的免疫组织化学染色
摘除上述实施例2、比较例3以及比较例4的小鼠的背皮肤组织利用10%福尔马林进行固定。之后,委托韩国CFC(韩国京畿道)进行苏木精和伊红 (Hematoxylin and eosin,H&E)染色后利用光学显微镜(日本奥林巴斯公司的IX71)进行观察,利用数码相机(日本奥林巴斯公司的DP71)拍照。
其结果,如图6的(B)部分所示,观察到比较例3-1)与未照射紫外线的比较例3-2)相比,皮肤表皮层变得增厚,在实施例2的情况下爱,因照射紫外线而变得增厚的表皮层的厚度显著减少。
实验例5.真皮层内胶原纤维(collagen fiber)的量及形态变化
除了进行马松三色(Masson's trichrome,M&T)染色之外,按照与上述
实验例4-(2)相同的方法进行。
其结果,如图7所示,比较例3-2)中,表皮下的真皮层中观察到了严重被苯胺蓝(aniline blue)染色的胶原纤维,但比较例3-1)中,胶原纤维只是轻微被苯胺蓝(anilineblue)染色。并且,实施例2相比比较例3-1),真皮层中胶原纤维的密度相对稠密,排列规则,但被苯胺蓝(aniline blue) 染色的程度更强,几近于比较例3-2)的状态。
实验例6.小鼠皮肤组织的基因表达变化
每0.1g上述实施例2、比较例3以及比较例4的小鼠的背皮肤添加1ml 的Trizol溶液并粉碎组织之后,在4℃、12000×g中进行10分钟的离心分离。将上层液移入新试管之后添加200ml的氯仿(chloroform)进行涡旋(vortex)。将这一过程反复两次之后,将上层液移入新试管之后,按照1:1比率添加异丙醇(isoprophanol)和上层液。大力摇晃10次之后室温放置15分钟之后,在12000×g、4℃条件下进行10分钟的离心分离之后去除上层液,在剩下的沉淀物中加入1ml的70%乙醇(ethanol)后在7500×g、4℃条件下进行 5分钟的离心分离。之后,将去除乙醇后装有RNA沉淀物的试管在室温下干燥15分钟之后,使用无核酸酶纯水(nuclease free water)使RNA微丸 (pellet)溶解。利用紫外/可见光分光光度计(UV/VIS spectrophotometer) (贝克曼库尔特公司的DU730)在260nm及280nm波长下测定所提取的 RNA试样的浓度,实施琼脂糖凝胶电泳(agarose gel electrophoresis)来确认RNA试样的完整性(integrity)。
将从背皮肤组织中提取的RNA试样作为对象,利用多聚胸腺嘧啶T重复寡核苷酸引物(oligo dT primer)和上标逆转录酶(superscript reverse transcriptase)(美国马里兰州盖瑟斯堡的GIBCO BRL公司)执行反转录 (reverse transcription)来合成cDNA。将通过反转录(Reverse transcription) 得到的cDNA作为模板(template),使用需要放大的基因cDNA的5’和3’侧翼序列(flanking sequence)作为引物(primer)来执行PCR(聚合酶链式反应)。此时使用的引物序列(primer sequence)如下表2。对放大的PCR 产物1μl利用1%琼脂糖凝胶(agarose gel)进行电泳来确认DNA带(band)。
表2
其结果,如图8所示,比较例3-1)与比较例3-2)相比,皮肤组织的 Col1α1、Col1α2以及Col3α1的表达明显减少,MMP-1a、MMP-1b、MMP-3 以及MMP-9基因的表达明显增加。另一方面,实施例2与比较例3-1)相比,Col1α1、Col1α2以及Col3α1的表达明显增加,MMP-1a、MMP-1b、 MMP-3以及MMP-9基因的表达明显减少。即,芥子酸能够增加皮肤组织的胶原蛋白蛋白质合成,阻碍胶原纤维的分解,从而抑制紫外线照射引起的皱纹形成。
并且,如图9所示,比较例3-1)相比未照射紫外线的比较例3-2),弹力蛋白的表达明显减少,实施例2与比较例3-1)相比,弹力蛋白的表达明显增加,而且与比较例4相比,也具有突出的效果。即,芥子酸能够发挥通过促进弹力蛋白表达来增强皮肤弹力的效果。
以下将对含有本发明的化合物的制剂例进行说明,但本发明并不是为了进行限定,而是详细说明。
制剂例1:散剂的制备
芥子酸:20mg
乳糖水合物:100mg
滑石:10mg
将上述成分混合后,填充到气密布制备散剂。
制剂例2:锭剂的制备
芥子酸:10mg
玉米淀粉:100mg
乳糖水合物:100mg
硬脂酸镁:2mg
将上述成分混合后,根据通常的锭剂的制备方法打锭制备锭剂。
制剂例3:胶囊剂的制备
芥子酸:10mg
微晶纤维素:3mg
乳糖水合物:14.8mg
硬脂酸镁:0.2mg
将上述成分混合后,根据通常的胶囊剂的制备方法填充到明胶胶囊来制备胶囊剂。
制剂例4:注射剂的制备
芥子酸:10mg
甘露醇:180mg
注射用灭菌蒸馏水:2974mg
磷酸一氢钠:26mg
将上述成分混合后,根据通常的注射剂的制备方法,按照每一安瓶(2mL)含有上述成分含量的方式制备。
制剂例5:液剂的制备
芥子酸:10mg
异性化糖:10mg
甘露醇:5mg
纯净水:适量
柠檬香:适量
将上述成分按照通常的制备方法在纯净水中加入各成分溶解之后,加入适量柠檬香之后加入纯净水调节为总量为100mL之后灭菌并填充到褪色瓶来制备液剂。
制剂例6:保健食品的制备
芥子酸:10mg
维生素混合物:适量
维生素A醋酸酯:70μg
维生素E:1.0mg
维生素B1:0.13mg
维生素B2:0.15mg
维生素B6:0.5mg
维生素B12:0.2μ
维生素C:10mg
生物素:10μg
烟酰胺:1.7mg
叶酸:50μg
泛酸钙:0.5mg
无机质混合物:适量
硫酸亚铁:1.75mg
氧化锌:0.82mg
碳酸镁:25.3mg
第一磷酸钾:15mg
第二磷酸钾:55mg
柠檬酸钾:30mg
碳酸钙:100mg
氯化镁:24.8mg
上述维生素及矿物质混合物的组成比是按照将比较适合于健康食品的成分按照优选的实施例混合组成的,但可以任意变形实施其配比,可以根据通常的健康食品制备方法混合上述成分之后制备颗粒,并根据通常的方法使用于健康食品组合物的制备。
制剂例7:健康饮料的制备
芥子酸:10mg
维生素C:15g
维生素E(粉末):100g
乳酸亚铁:19.75g
氧化锌:3.5g
烟酰胺:3.5g
维生素A:0.2g
维生素B1:0.25g
维生素B2:0.3g
纯净水:适量
根据通常的健康饮料制造方法将上述成分混合之后,在85℃条件下搅拌加热约1小时之后,将制得的溶液过滤并装入经灭菌处理的2l容器进行密封灭菌之后冷藏保管,之后用于制造本发明的健康饮料。
上述组成比是按照将比较适合于嗜好饮料的成分按照优选的实施例混合组成的,但可根据需要阶层或需要国家、使用用途等地域性、民族性爱好任意变形其配比来使用。
以下将对含有本发明的化合物的化妆品原料组合物进行说明,但本发明并不是为了进行限定,而是详细说明。
制造例1:营养化妆水(乳液)
本发明的芥子酸:2.0重量百分比
角鲨烯:5.0重量百分比
蜜蜡:4.0重量百分比
聚山梨酯60:1.5重量百分比
山梨坦倍半油酸酯:1.5重量百分比
液体石蜡:0.5重量百分比
辛酸/癸酸甘油酯:5.0重量百分比
甘油:3.0重量百分比
丁二醇:3.0重量百分比
丙二醇:3.0重量百分比
羧乙烯基聚合物:0.1重量百分比
三乙醇胺:0.2重量百分比
防腐剂、色素、香料:适量
纯净水:100重量百分比
上述配比是按照将比较适合于营养化妆水的成分按照优选的实施例混合组成的,但可任意变形其配比而使用,可根据通常的化妆品领域的制造方法制造。
制造例2:柔肤化妆水(爽肤水)
本发明的芥子酸:2.0重量百分比
甘油:3.0重量百分比
丁二醇:2.0重量百分比
丙二醇:2.0重量百分比
羧乙烯基聚合物:0.1重量百分比
PEG 12壬基苯醚:0.2重量百分比
聚山梨酯80:0.4重量百分比
乙醇:10.0重量百分比
三乙醇胺:0.1重量百分比
防腐剂、色素、香料:适量
纯净水:100重量百分比
上述配比是按照将比较适合于柔肤化妆水的成分按照优选的实施例混合组成的,但可任意变形其配比而使用,可根据通常的化妆品领域的制造方法制造。
制造例3:营养霜
本发明的芥子酸:2.0重量百分比
聚山梨酯60:1.5重量百分比
山梨坦倍半油酸酯:0.5重量百分比
PEG60氢化蓖麻油:2.0重量百分比
液体石蜡:10重量百分比
角鲨烯:5.0重量百分比
辛酸/癸酸甘油酯:5.0重量百分比
甘油:5.0重量百分比
丁二醇:3.0重量百分比
丙二醇:3.0重量百分比
三乙醇胺:0.2重量百分比
防腐剂:适量
色素:适量
香料:适量
纯净水:100重量百分比
上述配比是按照将比较适合于营养霜的成分按照优选的实施例混合组成的,但可任意变形其配比而使用,可根据通常的化妆品领域的制造方法制造。
制造例4:按摩霜
本发明的芥子酸:1.0重量百分比
蜜蜡:10.0重量百分比
聚山梨酯60:1.5重量百分比
PEG60氢化蓖麻油:2.0重量百分比
山梨坦倍半油酸酯:0.8重量百分比
液体石蜡:40.0重量百分比
角鲨烯:5.0重量百分比
辛酸/癸酸甘油酯:4.0重量百分比
甘油:5.0重量百分比
丁二醇:3.0重量百分比
丙二醇:3.0重量百分比
三乙醇胺:0.2重量百分比
防腐剂、色素、香料:适量
纯净水:100重量百分比
上述配比是按照将比较适合于按摩霜的成分按照优选的实施例混合组成的,但可任意变形其配比而使用,可根据通常的化妆品领域的制造方法制造。
制造例4:面膜
本发明的芥子酸:1.0重量百分比
聚乙烯醇:13.0重量百分比
羧甲基纤维素钠:0.2重量百分比
甘油:5.0重量百分比
尿囊素:0.1重量百分比
乙醇:6.0重量百分比
PEG12壬基苯醚:0.3重量百分比
聚山梨酯60:0.3重量百分比
防腐剂、色素、香料:适量
纯净水:100重量百分比
上述配比是按照将比较适合于面膜的成分按照优选的实施例混合组成的,但可任意变形其配比而使用,可根据通常的化妆品领域的制造方法制造。
制造例6:凝胶
本发明的芥子酸:0.5重量百分比
乙二胺醋乙酸钠(ethylendiaminesodium acetate):0.05重量百分比
甘油:5.0重量百分比
羧乙烯基聚合物:0.3重量百分比
乙醇:5.0重量百分比
PEG60氢化蓖麻油:0.5重量百分比
三乙醇胺:0.3重量百分比
防腐剂、色素、香料:适量
纯净水:100重量百分比
上述配比是将比较适合于凝胶的成分按照优选的实施例混合组成的,但可任意变形其配比而使用,可根据通常的化妆品领域的制造方法制造。
上述配比是将比较适合于化妆品原料组合物的成分按照优选的实施例混合组成的,但可适用于其他包含彩妆的各种用途的化妆品,根据其功效可用于制造薄薄地涂抹于人体的药剂即软膏,可根据需要阶层或需要国家、使用用途等地域性、民族性爱好任意变形其配比来使用。
Claims (8)
1.一种用于改善皮肤皱纹或增强弹力的化妆品原料组合物,其特征在于,包含芥子酸或其化妆品学上允许的盐作为有效成分。
2.根据权利要求1所述的用于改善皮肤皱纹或增强弹力的化妆品原料组合物,其特征在于,
上述芥子酸起到如下作用:
ⅰ)增加前胶原蛋白的分泌量,
ⅱ)减少基质金属蛋白酶基因的表达,
ⅲ)增加Ⅰ型胶原α1、Ⅰ型胶原α2或Ⅲ型胶原α1的表达,或者
ⅳ)增加弹力蛋白基因的表达。
3.根据权利要求1所述的用于改善皮肤皱纹或增强弹力的化妆品原料组合物,其特征在于,上述化妆品原料组合物还包含由维生素C、视黄酸、转化生长因子、动物胎盘来源的蛋白质、桦木酸以及小球藻提取物构成的组中的一种以上。
4.一种用于增强皮肤保湿、去除角质、抑制红斑或改善皮肤光老化的化妆品原料组合物,其特征在于,包含芥子酸或其化妆品学上允许的盐作为有效成分。
5.根据权利要求4所述的用于增强皮肤保湿、去除角质、抑制红斑或改善皮肤光老化的化妆品原料组合物,其特征在于,上述化妆品原料组合物抑制皮肤表皮层的增厚。
6.根据权利要求4所述的用于增强皮肤保湿、去除角质、抑制红斑或改善皮肤光老化的化妆品原料组合物,其特征在于,上述化妆品原料组合物抑制真皮与表皮交界处的胶原纤维网受损。
7.一种用于改善皮肤皱纹或增强弹性的保健食品组合物,其特征在于,包含芥子酸或其食品学上允许的盐作为有效成分。
8.一种用于增强皮肤保湿、去除角质、抑制红斑或改善皮肤光老化的保健食品组合物,其特征在于,包含芥子酸或其食品学上允许的盐作为有效成分。
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