CN104717957A - 包含藜芦酸或其盐作为有效成分的皮肤状态改善用组合物 - Google Patents
包含藜芦酸或其盐作为有效成分的皮肤状态改善用组合物 Download PDFInfo
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- CN104717957A CN104717957A CN201380052911.2A CN201380052911A CN104717957A CN 104717957 A CN104717957 A CN 104717957A CN 201380052911 A CN201380052911 A CN 201380052911A CN 104717957 A CN104717957 A CN 104717957A
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- skin
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Abstract
本发明涉及包含藜芦酸(veratric acid)及其盐作为有效成分的皮肤状态改善用化妆品、药学及食品组合物。本发明的组合物增大胶原蛋白(collagen)合成,并抑制胶原酶(collagenase)的活性,从而具有优秀的皱纹改善功效。并且,即使在照射紫外线的情况下,也增进因紫外线而减少的胶原蛋白的量,并减少因紫外线而增进活性的胶原酶的活性,从而具有优秀的改善皱纹功效。并且,促进毛囊真皮乳头细胞的增殖,并抑制胰岛素样生长因子-1(IGF-1)的分泌促进及转化生长因子-β1(TGF-beta1)的合成,从而具有促进育发及预防脱发的功效。并且,由于本发明的组合物来源于作为天然物质的植物,因而对人体无害,无皮肤副作用,从而能够安全地适用于化妆品、药学及食品组合物。
Description
技术领域
本发明涉及包含藜芦酸或药学上能够接受的其盐的组合物,更详细地,涉及产品稳定性优秀,能够在对皮肤无副作用的情况下安全地使用,并对防紫外线、改善皱纹效果、促进生发及预防脱发优秀的组合物。
背景技术
因环境污染、汽车尾气等,激素不均衡现象越来越严重。由此,脱发发生率变高,发生年龄也变低,导致皮肤免疫系统不协调,从而发生各种皮肤疾病。因此,深度研究开发可有效解决除了美观之外,还成为严重的社会问题的各种现象,例如,由内源性衰老和紫外线引起的皱纹、免疫不均衡及脱发等现象的物质。
皱纹因皮肤老化而发生,老化的皮肤是指与老化的过程相关的自然变化的总体。皮肤老化大致区分为两种,即,区分为在整个皮肤表面表示与年龄相关的皮肤功能、结构或形状变化的生理老化和由紫外线引起的光老化。
若皮肤进行老化,则明显发生真皮的变化。70岁以后发生的真皮萎缩是代表性的老化现象。真皮的变化是因成纤维细胞(fibroblast)的数量和它们的合成能力的减少而由细胞外基质中具有大分子量的物质变化而发生。其具体变化可例举胶原束的分离、糖胺聚糖的合成减少、胶原蛋白和弹性纤维(elastin)的数量和直径的减少、胶原蛋白和弹性纤维的粉碎及血管的膨胀等。一般,在皮肤的水分含量、胶原蛋白含量及对外部环境的免疫作用能力等各种复合因素中,对皱纹的形成影响最大的是作为减少胶原蛋白的生成量和胶原蛋白含量的胶原蛋白分解酶的胶原酶的表达量和活性。
作为用于预防如上所述的皱纹的方法,具有剥皮术、肉毒杆菌及填充剂等,在上述剥皮术中,使用抑制活性氧的发生来预防细胞的老化的抗氧化剂,或者使用羟基酸(乙醇酸)、β羟基酸(水杨酸)等的化学药品来剥皮肤,从而改善形成于皮肤的薄的皱纹,上述肉毒杆菌向形成有皱纹的面部肌肉注射规定量的肉毒杆菌毒素来改善皱纹,上述填充剂由透明质酸(hyalulonic acid)等的物质填充皱纹部位来缓解皱纹。
就上述方法而言,皱纹改善效果微弱,或者使用过多的情况下,发生副作用,并存在稳定性及安全性问题,因而需要开发来源于天然物质,从而副作用少、安全,且皱纹改善效果优秀的组合物。
脱发是指应正常存在毛发的部位没有毛发的状态,一般意味着头皮的终毛脱落。脱发的原因由遗传原因和作为男性激素的雄激素(androgen)作为主要因子而发生。与脱发相关的组织有皮脂腺(sebaceousgland)、表皮和毛囊的角化细胞(keratinocyte)、真皮乳头细胞(dermal papilla cell)、汗腺(sweat gland)、毛根鞘(root sheath of the scalp hair follicle)等。
据报告,脱发部位组织比普通毛发部位组织,5-α还原酶(5-αreductase)的活性更高。众所周知,5-α还原酶使睾酮(testosterone)还原为双氢睾酮(DHT,dehydrotestosterone),在男性的第二性征中,睾酮参与骨骼肌的增加、精子的形成等,双氢睾酮参与青春痘、皮脂的增加、脱发及前列腺肥大症等的相应组织中(J.Invest.Dermatol.1995;105(2):209-14)。因此,正在进行很多寻找如下物质的研究,上述物质为抑制参与从睾酮还原为双氢睾酮的反应的生物酶,即,5-α还原酶的活性,来治疗脱发症的药理活性物质。
以往,5-α还原酶抑制剂有非那雄胺(finasteride)之类的化合物,改变给药容量来使用为前列腺治疗剂及防脱发剂。非那雄胺是在便利性和功效方面优秀的药,但由于用来制备非那雄胺的试剂价格高或具有剧毒性,因而存在制备成本负担重或污染环境的问题,尤其,由于难以除去副产物,因而目的产物的纯度下降,或者由于使用容易被水分抑制的试剂或活性衍生物,因而难以进行大量生产。而且,类固醇激素本身也对正常的生理活性起到重要的作用,因而在人为阻断类固醇激素的生成本身的情况下,有可能导致各种问题。
因而,需要开发比抑制5-α还原酶活性的物质,更促进真皮乳头细胞的增殖本身,或者促进帮助增殖的生长因子的分泌,并抑制作为如上所述的酶的下游信号转导物质的TGF-β1生物合成的物质。
作为来源于天然物质的改善皮肤皱纹、预防脱发及育发方法,在韩国公开特许第0868784号中公开了含有网盖红褶伞液体培养提取物的皱纹改善化妆品组合物,在韩国公开特许第2007-0123712号中公开了包含红毛丹及荔枝提取物作为有效成分的皮肤老化防止用及皮肤皱纹改善用化妆品组合物。并且,在韩国公开特许第2010-0119226号中公开了包含海藻类提取物的防止脱发或促进育发的组合物,在韩国公开特许第2004-0097416号中公开了含有猕猴桃提取物的脱发及脂溢性皮肤症状预防及改善用保健食品。并且,在韩国公开特许第2011-0078678号中公开了包含异黄酮苷元的皱纹改善用化妆品组合物,在韩国公开特许第2008-0069816号中公开了含有普洱茶提取物的皮肤皱纹及弹性改善组合物。并且,在韩国公开特许第2009-0021963号中公开了以2-甲基-庚-2-烯(2-methyl-hept-2ene)衍生物及氧化芳樟醇(linalool oxide)的一种以上作为有效成分的防脱发及促进毛发生长的组合物,在韩国公开特许第2012-0039384号中公开了含有类黄酮的生发及育发组合物。
但是,到目前为止,还没有公开用于利用如本发明中的藜芦酸(veratric acid)或其盐来防紫外线、改善皮肤皱纹、促进生发及预防脱发的方法。
对此,本发明人为了利用天然物质来开发对人体无害,安全性和稳定性优秀,且效果突出的来源于植物的皮肤状态改善用组合物而努力,其结果发现以藜芦酸或其盐作为有效成分的组合物对防紫外线效果、改善皮肤皱纹、促进生发及预防脱发非常有效,从而完成了本发明。
发明内容
技术问题
本发明的一目的在于,提供包含藜芦酸(veratric acid)或化妆学上能够接受的其盐作为有效成分的皮肤状态改善用化妆品组合物。
本发明的再一目的在于,提供包含藜芦酸(veratric acid)或药学上能够接受的其盐作为有效成分的皮肤状态改善用药学组合物。
本发明的另一目的在于,提供包含藜芦酸(veratric acid)或食品学上能够接受的其盐作为有效成分的皮肤皱纹改善及预防用食品组合物。
技术方案
作为一实施方式,本发明涉及包含藜芦酸或化妆学上能够接受的其盐作为有效成分的皮肤状态改善用化妆品组合物。
在本发明中,上述藜芦酸(veratric acid)具有3,4-二甲氧基苯甲酸(3,4-dimethoxybenzoic acid)的IURAC名的物质,其结构如以下化学式1。
化学式1:
上述藜芦酸可从风铃木(Tabebuia impetiginosa)、留兰香(Mentha spicata)等中提取、分离及纯化,并具有抗氧化效果和降压效果,据报告,是一种可通过抑制3-羟基-3-甲基戊二酰-辅酶A(HMG-CoA,3-hydroxy-3-methylglutaryl-coenzyme A)还原酶来改善心血管疾病及高脂肪血症的候选物质(Eur J Pharmacol.2011;J Cardiovasc Pharmacol.2012;Mol Cell Biochem.2012)。
在上述植物中,利用溶剂来提取藜芦酸的情况下,可将水、碳原子数为1~4的无水或含水低级醇(甲醇、乙醇、丙醇、丁醇等)、上述低级醇和水的混合溶剂、丙酮、乙酸乙酯(ethyl acetate)、氯仿(chloroform)或1,3-丁二醇(1,3-butylene glycol)作为提取溶剂来获取。优选地,利用甲醇、乙醇或丁醇水溶液来提取,更优选地,利用丁醇水溶液来提取。
并且,上述藜芦酸除了利用上述提取溶剂来提取的方法之外,还可经过通常的纯化过程来获取。例如,还可通过分馏物获取藜芦酸,通过利用具有规定分子量截止值的超滤膜的分离、利用各种色谱法(为了基于大小、电荷、疏水性或亲和性的分离而制备)的分离等、追加实施的各种纯化方法获取上述分馏物。
在本发明中,作为上述藜芦酸的化妆学上能够接受的盐,由化妆学上能够接受的游离酸(free acid)形成的酸加成盐或由碱形成的金属盐有用。作为一例,游离酸可使用无机酸和有机酸。无机酸可使用盐酸、硫酸、溴酸、亚硫酸或磷酸等,有机酸可使用柠檬酸、醋酸、马来酸、富马酸、葡萄糖酸、甲磺酸等。并且,金属盐可使用碱金属盐或碱土金属盐、钠、钾、钙盐。但并不局限于此。
在本发明中,改善皮肤状态是指防紫外线、改善皱纹、促进生发及预防脱发。
在本发明中,包含藜芦酸或化妆学上能够接受的其盐的化妆品组合物保护成纤维细胞,促进胶原蛋白的合成,并抑制胶原酶的活性,来呈现优秀的皮肤皱纹改善功效。
一个具体实施中,利用角质形成细胞(HaCaT)和成纤维细胞(fibroblast)来照射户外紫外线(UVB),并观察根据藜芦酸浓度的细胞生存率,其结果,确认了藜芦酸的浓度越高,细胞的生存率越高。上述结果表明藜芦酸可呈现防紫外线效果。
再一个具体实施中,利用人的正常成纤维细胞来观察根据藜芦酸浓度的胶原蛋白合成增加率,其结果确认了藜芦酸的浓度越高,胶原蛋白合成增加率越高。上述结果表明藜芦酸可通过合成胶原蛋白来呈现改善皱纹的效果。
另一个具体实施中,利用人类的正常成纤维细胞来照射户外紫外线,并观察根据藜芦酸浓度的胶原蛋白合成量及胶原酶活性,其结果确认了因照射户外紫外线而使活性增加的胶原酶活性减少,且藜芦酸的浓度越增加,因照射户外紫外线而减少的胶原蛋白合成就越增加。上述结果表明藜芦酸可呈现皱纹预防效果。
还一个具体实施中,对人类皮肤使用包含藜芦酸的化妆品来观察皮肤弹性,其结果确认了包含在化妆品的藜芦酸的含量越高,皮肤弹性就越增加。上述结果表明藜芦酸可呈现皱纹改善效果。
在本发明中,上述包含藜芦酸或化妆学上能够接受的其盐的组合物抑制毛囊真皮乳头细胞的增殖作用、调节毛发的生长循环的生长因子IGF-1(Insulin-like growth factor-1)的分泌促进作用及构成毛发的蛋白质的合成抑制因子TGF-β1(Tranforming growth factor-β1)的合成,来呈现促进生发及预防脱发的功效。
一个具体实施中,利用人类的毛囊真皮乳头细胞来观察根据藜芦酸浓度的细胞的增殖率,其结果表明藜芦酸的浓度越高,毛囊真皮乳头细胞的增殖率越高,从而确认到藜芦酸使毛囊真皮乳头细胞增殖来呈现促进生发效果。并且,对脱发症患者的脱发部位适用按不同浓度包含藜芦酸的生发剂6个月,其结果表明从1个月至2个月开始就出现新的毛发,并确认到在适用藜芦酸的浓度高的生发剂的实验组中,效果更好。因而,藜芦酸呈现促进生发功效。
再一个具体实施中,利用人类的毛囊真皮乳头细胞来观察根据藜芦酸浓度的IFG-1及TGF-β1的合成量,其结果确认了藜芦酸的浓度越增加,IGF-1的分泌量越增加,TGF-β1的分泌量越减少。上述结果表明藜芦酸可呈现防脱发效果。
本发明的化妆品组合物除了作为具有防紫外线、改善皱纹、促进生发及预防脱发功效的有效成分的藜芦酸或化妆学上能够接受的其盐之外,还包含化妆品组合物中通常所利用的成分。例如,包含作为抗氧化剂、稳定剂、增溶剂、维生素、颜料及香料之类的化妆品接受的通常的辅助剂及载体。
上述化妆品组合物可剂型化为溶液、悬浮液、乳浊液、糊剂、凝胶、乳霜、乳液、粉饼、肥皂、含有表面活性剂的洁面霜、油、粉末粉底、乳浊液粉底、蜡粉底及喷剂等,但并不局限于此。更详细地,化妆品组合物可制备为营养霜、收敛化妆水、柔软化妆水、乳液、精华素、洗发露、营养乳或按摩霜的剂型。
在上述化妆品组合物的剂型为糊剂、乳霜或凝胶的情况下,作为载体成分,可包含动物油、植物油、蜡、石蜡、淀粉、黄芪、纤维素衍生物、聚乙二醇(polyethylene glycol)、硅、膨润土、二氧化硅、滑石或氧化锌等。
上述化妆品组合物的剂型为粉饼或喷剂的情况下,作为载体成分,可包含乳糖、滑石、二氧化硅、氢氧化铝、硅酸钙或聚酰胺粉,尤其,在喷剂的情况下,还可包含氯氟烃(chlorofluorohydrocarbon)、丙烷/丁烷或二甲醚(dimethyl ether)之类的推进剂。
上述化妆品组合物为溶液或乳浊液的情况下,作为载体成分,可利用溶剂、增溶剂或乳化剂,例如,可包含水、乙醇、异丙醇(isopropanol)、碳酸乙酯(ethyl carbonate)、乙酸乙酯(ethyl acetate)、苯甲醇(benzyl alcohol)、苯甲酸苄酯(benzyl benzoate)、丙二醇(propylene glycol)、1,3-丁基乙二醇油(1,3-butyl glycol oil)、甘油脂肪酯(glycerol-aliphatic ester)、聚乙二醇或山梨糖醇脂肪酯(sorbitan-aliphatic ester)等。
上述化妆品组合物的剂型为悬浮液的情况下,作为载体成分,可包含水、乙醇或丙二醇之类的液相稀释剂、乙氧基化异硬脂醇(ethoxylated isostearyl alcohol)、聚氧乙烯山梨醇酯(polyoxyethylene sorbitol ester)或聚氧乙烯山梨糖醇酐酯(polyoxyethylene sorbitan ester)之类的悬浮剂、微晶纤维素、甲基氢氧化铝(aluminum metahydroxide)、膨润土、琼脂或黄芪等。
上述化妆品组合物的剂型为含有表面活性剂的洁面霜的情况下,作为载体成分,可包含脂肪族醇硫酸盐、脂肪族醇醚硫酸盐、磺基琥珀酸单酯、羟乙基磺酸盐、咪唑啉衍生物、甲基牛磺酸盐、肌氨酸盐、脂肪酸酰胺醚硫酸盐、烷基酰胺甜菜碱、脂肪族醇、脂肪酸甘油酯、脂肪酸二乙醇酰胺、植物油、羊毛脂衍生物或乙氧基化甘油脂肪酸酯等。
在上述化妆品组合物中,相对于组合物的总重量,可包含0.00001至15重量百分比的上述藜芦酸或化妆学上能够接受的其盐,优选地,可包含0.0001至10重量百分比,更优选地,可包含0.0001至5重量百分比。在藜芦酸的重量小于0.00001重量百分比的情况下,防紫外线、改善皱纹、促进生发及预防脱发的功效微弱,在藜芦酸的重量大于15重量百分比的情况下,随着含量增加而增加的效果非常微弱,并存在不能确保剂型的稳定性的问题。
在使用包含上述藜芦酸或化妆学上能够接受的其盐作为有效成分的化妆品组合物的情况下,可增加成纤维细胞的保护作用、皮肤的胶原蛋白的合成,或者抑制胶原酶的活性,来呈现皮肤皱纹改善功效。并且,可抑制毛囊真皮乳头细胞的增殖促进、作为毛发生长因子的IGF-1的分泌促进及作为毛发抑制因子的TGF-β1的分泌,来呈现促进生发及预防脱发的功效。
作为再一实施方式,本发明涉及包含藜芦酸或药学上能够接受的其盐作为有效成分的皮肤状态改善用药学组合物。
与本发明的藜芦酸的结构、提取法及功效等相关的内容如上所述。
并且,在本发明中,改善皮肤状态如上所述。
在本发明中,作为上述藜芦酸的药学上接受的盐,由药学上能够接受的游离酸(free acid)形成的酸加成盐或由碱形成的金属盐有用。作为一例,游离酸可使用无机酸和有机酸。无机酸可使用盐酸、硫酸、溴酸、亚硫酸或磷酸等,有机酸可使用柠檬酸、醋酸、马来酸、富马酸、葡萄糖酸、甲磺酸等。并且,金属盐可使用碱金属盐或碱土金属盐、钠、钾、钙盐。但并不局限于此。
并且,本发明的药学组合物除了作为具有防紫外线、改善皱纹、促进生发及预防脱发功效的有效成分的藜芦酸或药学上能够接受的其盐之外,还包含药学组合物中通常所利用的成分。例如,本发明的组合物可包含润滑剂、润湿剂、甜味剂、香味剂、乳化剂、悬浮剂、保鲜剂及载体。
适合作为上述药学组合物的载体,可例举乳糖、葡萄糖、蔗糖、山梨糖醇、甘露醇、淀粉、阿拉伯胶、磷酸钙、藻酸盐、明胶、硅酸钙、微晶纤维素、聚乙烯吡咯烷酮(polyvinylpyrrolidone)、纤维素、水、糖浆、甲基纤维素、羟基苯甲酸甲酯(methyl hydroxybenzoate)、羟基苯甲酸丙酯(propyl Hydroxybenzoate)、滑石、硬脂酸镁及矿物油等,但并不局限于此。例如,在Remington's Pharmaceutical Sciences(19th ed.,1995)中详细记载了适当的药学上接受的载体及制剂。
上述药学组合物能够以口服或非口服的方式进行给药。优选地,能够以非口服的方式进行给药,更优选地,能够以基于涂敷的局部给药(topical application)的方式适用。
上述药学组合物以药学上有效的量进行给药。在本发明中,药学上有效的量是指对以可适用于医学治疗或预防的合理的受益/危险比率治疗或预防疾病充分的量,有效容量水平可根据疾病种类及其重症度、药物的活性、患者的年龄、体重、健康、性别、患者对药物的敏感度、所使用的特定提取物的给药时间、给药途径及排放率、治疗期间、包含与所使用的特定提取物配合或同时所使用的药物的因素及在医学上熟知的其他因素而决定。一般,以成人为基准,可将0.001至100mg/kg的容量以一天1次或数次的方式进行给药。并且,在外用剂的情况下,优选地,以成人为基准,以1.0至3.0ml的量,以一天1次至5次涂敷的方式继续给药1个月以上。但本发明的范围并不局限于上述给药量。
上述药学组合物根据本发明所属技术领域的普通技术人员可容易实施的方式,来与药学上接受的载体和/或赋形剂一同进行制剂化,从而能够以单位容量形态制备或者以內入于大容量容器内的方式制备。此时,剂型可根据治疗目的剂型化为药剂学领域中通常的制剂,例如,可剂型化为片剂、胶囊剂、散剂、颗粒、悬浮剂、乳剂、糖浆剂、乳浊剂、硬膏剂、软膏剂、喷剂、油剂、凝胶剂、酒精剂、酊剂、浴剂、搽剂、乳液剂、贴剂、垫剂、乳霜剂等。并且,优选地,可用作以直接涂敷于患处的局部给药为目的的皮肤外用剂,在此情况下,优选为软膏剂、乳液剂、喷剂、凝胶等的形态。并且,上述皮肤外用剂可包含在能够直接适用于治疗部位的载体(support base)或基质,基体例如包括纱布或绷带。
在上述药学组合物中,相对于组合物的总重量,包含0.00001至15重量百分比的上述藜芦酸或药学上能够接受的其盐,优选地,包含0.0001至10重量百分比,更优选地,包含0.0001至5重量百分比。在藜芦酸的重量小于0.00001重量百分比的情况下,防紫外线、改善皱纹、促进生发及预防脱发的功效微弱,在藜芦酸的重量大于15重量百分比的情况下,随着含量增加而增加的效果非常微弱,并存在不能确保剂型的稳定性的问题。
在使用包含上述藜芦酸或药学上能够接受的其盐作为有效成分的药学组合物的情况下,可增加成纤维细胞的保护作用、皮肤的胶原蛋白的合成,或者抑制作为分解胶原蛋白的酶的胶原酶的活性,来呈现皮肤皱纹改善功效。并且,可抑制毛囊真皮乳头细胞的增殖促进、IGF-1的分泌促进及TGF-β1的分泌,来呈现促进生发及预防脱发的功效。
作为另一实施方式,本发明涉及包含藜芦酸或食品学上能够接受的其盐作为有效成分的皱纹改善及预防用食品组合物。
与上述藜芦酸的结构、提取法及功效等相关的内容如上所述。
在本发明中,作为上述藜芦酸的食品学上接受的盐,由食品学上能够接受的游离酸(free acid)形成的酸加成盐或由碱形成的金属盐有用。作为一例,游离酸可使用无机酸和有机酸。无机酸可使用盐酸、硫酸、溴酸、亚硫酸或磷酸等,有机酸可使用柠檬酸、醋酸、马来酸、富马酸、葡萄糖酸、甲磺酸等。并且,金属盐可使用碱金属盐或碱土金属盐、钠、钾、钙盐。但并不局限于此。
一个具体实施中,观察对照射紫外线(UV)的雌性裸鼠以口服的方式进行给药的情况的效果,其结果确认到在将藜芦酸给药的组中具有预防皱纹效果。
在本发明的组合物由食品组合物制备的情况下,除了藜芦酸或食品学上能够接受的其盐之外,还可包含在制备食品时通常所添加的成分作为有效成分。例如,包含蛋白质、碳水化合物、脂肪、营养素、调味剂及香味剂。上述碳水化合物为葡萄糖及果糖等的单糖、麦芽糖、蔗糖及低聚糖等的二糖、糊精及环糊精等的多糖或木糖醇、山梨糖醇、赤藓糖醇等的糖醇。上述香味剂可使用天然香味剂(奇异果甜蛋白、甜叶菊提取物(例如,蛇菊苷A,甘草甜素等))或合成香味剂(糖精、阿斯巴甜等)。
上述食品的种类没有特别的限制。可添加上述藜芦酸或食品学上能够接受的其盐的食品例如有肉类、香肠、面包、巧克力、糖果类、小吃类、饼干类、比萨、方便面、其他面类、口香糖类、包含冰淇淋类的乳制产品、各种汤、饮料、茶、口服液、酒精饮料及维生素复合剂等,包括通常意义上的所有保健食品。
在本发明的保健食品为保健饮料的情况下,与通常的饮料一样,还可包含各种香味剂或天然碳水化合物等成分。上述天然碳水化合物可使用葡萄糖、果糖之类的单糖、麦芽糖、蔗糖之类的二糖以及糊精、环糊精之类的天然甜味剂或糖精、阿斯巴甜之类的合成甜味剂等。一般,上述天然碳水化合物的比率为本发明的每100ml保健饮料中占约0.01至0.4g,优选地,占约0.02至0.03g。
并且,上述保健食品除了上述成分之外,还可包含各种营养剂、维生素、电解质、风味剂、着色剂、果胶酸及其盐、海藻酸及其盐、有机酸、保护性胶体增稠剂、pH调节剂、稳定剂、防腐剂、甘油、乙醇、用于碳酸饮料的碳酸化剂等。除此之外,本发明的保健食品可包含天然果汁、果汁饮料及用于制备蔬菜饮料的果肉。这种成分可单独或组合使用。这种添加剂的比率不太重要,但一般在本发明的每100重量份的保健食品中,在0.01至0.1重量份的范围内进行选择。
在上述食品组合物中,相对于组合物的总重量,可包含0.00001至15重量百分比的上述藜芦酸或食品学上能够接受的其盐,优选地,可包含0.0001至10重量百分比,更优选地,可包含0.0001至5重量百分比。在藜芦酸的重量小于0.00001重量百分比的情况下,皱纹改善及预防功效微弱,在藜芦酸的重量大于15重量百分比的情况下,随着含量增加而增加的效果非常微弱,并存在不能确保剂型的稳定性的问题。
在使用包含上述藜芦酸或食品学上能够接受的其盐作为有效成分的食品组合物的情况下,可增加皮肤的胶原蛋白的合成,或者抑制胶原酶的活性,来呈现皮肤皱纹改善功效。
再一个具体实施中,利用藜芦酸来进行皮肤累积刺激性实验,其结果表明藜芦酸作为天然物质对人体无害。因此,本发明的藜芦酸几乎没有毒性及副作用,因而也可长时间放心使用,尤其,可安全地适用于如上所述的化妆品、药学及食品组合物。
本发明的组合物可向个体给药来改善个体的皮肤状态。
在本发明中,术语“个体”用于改善或预防皮肤的不良状态,例如,皱纹、皮肤黑化及脱发等,是指可通过将本发明的组合物进行给药来达成这些目的的包括人类的哺乳动物,例如,狗、猪、马及羊。
在本发明中,术语“给药”是指利用适当的方法来向个体导入规定的物质的过程,作为本发明的组合物的给药途径,只要可达到目的组织,就可通过任何普通的途径以口服或口服的方式进行给药。并且,在本发明的组合物中,可借助目标对象,例如可向细胞移动的任何装置来将活性物质进行给药。
当然,本发明所属技术领域的普通技术人员能够通过上述内容及以下记载的实施例等理解上述的皮肤状态或异常状态可通过向本发明的组合物的个体内进行给药来达成改善、好转、预防或治疗等目的。
有益的效果
如上所述,本发明的包含藜芦酸及其盐作为有效成分的化妆品、药学及食品组合物具有防紫外线、改善皱纹、促进生发或预防脱发的功效。并且,因无细胞毒性及皮肤副作用,能够安全地适用于化妆品、药学及食品组合物。
附图说明
图1表示根据藜芦酸浓度的毛囊真皮乳头细胞的增殖率。
图2表示根据藜芦酸浓度的IGF-1的分泌率。
图3表示根据藜芦酸浓度的TGF-β1的抑制率。
图4表示在向人的正常成纤维细胞照射长波紫外线(UVA)的情况下,根据藜芦酸浓度的胶原蛋白的合成率。
图5表示在向成纤维细胞照射UVB的情况下,根据藜芦酸浓度的细胞生存率。
图6表示在向角质形成细胞照射UVB的情况下,根据藜芦酸浓度的细胞生存率。
图7表示在向人类的正常成纤维细胞照射UVB的情况下,根据藜芦酸浓度的胶原酶活性。
具体实施方式
以下,通过实施例更加详细地说明本发明。但这些实施例仅用于更具体地说明本发明,根据本发明的要旨,本发明的范围并不局限于这些实施例,这对于本发明所属技术领域的普通技术人员来说是显而易见的。
实施例1:生发剂的制备
以下列表1的成分含量将含有藜芦酸的生发剂制备成水凝胶基体形态。具体地,将水相的蒸馏水加热至70℃后,将油相的防腐剂和增稠剂加热至70℃,利用均质混合机(日本特殊电装基卡(Tokushu Kika)公司)来进行搅拌并乳化。在乳化结束后,将上述溶液冷却至45℃,并相对于组合物的总重量,添加0.1及1.0重量百分比的藜芦酸进行分散,之后,冷却至30℃。在表1中,实验组1及实验组2分别为包含藜芦酸的生发剂组合物。
表1
实施例2:藜芦酸的促进生发及预防脱发的测定
2-1.体外(in vitro)条件下的藜芦酸的促进生发及预防脱发的测定
在添加了10%胎牛血清(FBS)的具有DMEM培养基的(美国吉毕科(Gibco.USA))6孔微板中以每孔有3×105个细胞的方式接种人类毛囊真皮乳头细胞(Hair follicle dermal papillar cell;Application,Inc.美国(USA))后,在37℃温度下,利用浓度为5%的CO2培养器来培养了24小时。第二天,利用除去了血清的培养液来清洗一次,之后,更换为除去了血清的培养液。之后,处理0、10及100μM的藜芦酸,并培养了72小时。之后,利用除去了血清的培养液来清洗一次,之后,更换为具有10%的3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)的无血清培养基,并反应3小时后,测定吸光度(O.D.)值,并换算为百分比。在图1中示出了其结果。
根据实验结果,观察到藜芦酸的浓度越增加,人类毛囊真皮乳头细胞数越增加。
2-2.体内(in vivo)条件下的藜芦酸的促进生发及预防脱发的测定
将因毛囊萎缩而头皮光滑的脱发秃顶患者、典型的男性脱发症患者及急性斑秃症患者等40名(年龄:40岁到60多岁)分为4组,并一组分配10名。将在上述实施例1中制备的生发剂以一天两次,每次3㏄的方式适用于脱发症患者的脱发部位六个月。每一个月观察脱发及育发状态。使用市场上销售的米诺地尔(韩美药品)作为比较组,并只使用50%的乙醇作为对照组。在表2中示出了其结果。
判定基准如下:
4:效果很好=有新的毛发
3:有中等程度的效果=有新的毛发(毳毛)
2:有点效果=脱发数减少
1:没有效果=完全没有变化
表2
— | 对照组 | 比较组 | 实验组1 | 实验组2 |
功效/效果 | 1.6 | 4.3 | 2.7 | 3.4 |
如上述表2,在使用包含本发明的藜芦酸的生发剂的脱发症患者中,从治疗1个月或2个月开始出现毳毛和硬毛的新的毛发,从治疗6个月开始在80%以上的患者中出现了生发效果。并且,观察到新的毛发持续生长,并未发现脱发现象。
实施例3:藜芦酸的生长因子IGF-1的分泌促进效果的测定
在24孔微板中以每孔有7.5×104个细胞的方式接种人类毛囊真皮乳头细胞(Application,Inc.,美国(USA))后,在37℃温度下,利用浓度为5%的CO2培养器来培养了24小时。之后,将培养液更换为未添加抗生剂的DMEM培养液后培养了16小时。之后,在各个孔中处理0、10、100μM藜芦酸,再培养48小时,之后,只收集了细胞培养液。
利用生长因子IGF-1测定试剂盒(DG100,R&D systems,美国(USA))来测定了生长因子IGF-1(Insulin-like growth factor-1)的分泌量。在图2中示出了其结果。
根据实验结果可知,藜芦酸在人类毛囊真皮乳头细胞中以浓度依赖性的方式增加了IGF-1的生成。
实施例4:藜芦酸的TGF-β1的生成抑制效果的测定
在24孔微板中以每孔有7.5×104个细胞的方式接种人类毛囊真皮乳头细胞(Application,Inc.,美国(USA))后,在37℃温度下,利用浓度为5%的CO2培养器来培养了24小时。之后,将培养液更换为未添加抗生剂的DMEM培养液后培养了16小时。之后,在各个孔中处理0、1、10及100μM的TGF-β1(Transforming growth factor-β1),再培养48小时,48小时之后,只收集了细胞培养液。
利用TGF-β1测定试剂盒(ab100647,abcam,美国(USA))来测定了TGF-β1的分泌量。在图3中示出了其结果。
实验结果表明,藜芦酸呈现优秀的TGF-β1生成抑制效果,藜芦酸可抑制TGF-β1的表达来呈现生发促进功效。
实施例5:藜芦酸的皱纹改善效果的测定
在具有DMEM培养基的24孔微板中以每孔有2×105个细胞的方式接种人类正常成纤维细胞(human normal fibroblast,亚洲大学(Ajou University)皮肤科),并在浓度为5%的CO2培养器中,以37℃温度培养了24小时。之后,除去各个孔的培养基,并处理0、1、10及100μM浓度的藜芦酸,再培养24小时,之后,收集细胞培养基,并制备了样品。
对于制备的上述样品的胶原蛋白的合成量,利用胶原蛋白测定试剂盒(原骨胶原类IC-肽酶免疫测定试剂盒(Procollagen Type I C-peptide EIA kit)(MK101),日本京都TAKARA)来测定了原骨胶原(procollagen)类I C-肽(Type I C-peptide:PICP)的量。在图4中示出了其结果。
实验结果表明,藜芦酸增加了胶原蛋白的合成,藜芦酸的浓度越增加,人类正常成纤维细胞的胶原蛋白的合成率上升,从而表示具有浓度依赖性的倾向。
实施例6:藜芦酸的防紫外线效果的测定
在由青霉素-链霉素及血清构成的DMEM基本培养液中培养了由紫外线诱导的角质形成细胞(HaCaT)和成纤维细胞(fibroblasts)。在24孔微板中以每孔有约1×105个细胞的方式接种后,在37℃温度下,在浓度为5%的CO2培养器中,培养至约80%以上的细胞附着于孔底面。在利用紫外线来照射24小时之前,添加了0、1、10及100μM的藜芦酸。将培养液换为磷酸盐缓冲液(PBS),并照射了紫外线(UVB20mJ/cm2)。在照射紫外线后,换为由青霉素-链霉素及血清构成的DMEM基本培养液,再培养48小时,在培养最后一天,利用细胞毒性(MTT assay;ATCCTM,30-1010K)调查来测定了细胞生存力。在图5和图6中示出了其结果。
实验结果表明,藜芦酸抑制由紫外线诱导的角质形成细胞和成纤维细胞的细胞死亡,藜芦酸的浓度越增加,细胞死亡的抑制率越增加,从而表示具有浓度依赖性的倾向。
实施例7:藜芦酸的胶原蛋白分解酶的抑制效果的测定
在具有DMEM培养基的24孔微板中以每孔有2×105个细胞的方式接种人类正常成纤维细胞(human normal fibroblast,亚洲大学皮肤科),并在浓度为5%的CO2培养器中,以37℃温度培养了24小时。之后,在各个孔中除去培养基,并处理0、1、10及100μM浓度的藜芦酸,再培养24小时,并收集细胞培养基,从而制备了样品。
对于制备的上述样品的胶原蛋白分解酶抑制效果的测定,利用胶原蛋白分解酶测定试剂盒(MMP-1ELISA kit(QIA55),Merck,德国(Germany))来测定了胶原蛋白分解酶量。在图7中示出了其结果。
实验结果表明,藜芦酸减少了人类正常成纤维细胞的胶原蛋白分解酶的生成量,藜芦酸的浓度越增加,人类正常成纤维细胞的胶原蛋白分解酶的生成量越减少,从而表示具有浓度依赖性的倾向。
实施例8:含有藜芦酸的化妆品的皱纹改善效果的测定
8-1.含有藜芦酸的营养霜的制备
制备了具有下列表3的成分及含量的含有藜芦酸的营养霜。具体地,将水相的纯化水、三乙醇胺(triethanolamine)及丙二醇加热至70℃并溶解后,添加将油相的脂肪酸、油性成分,乳化剂及防腐剂加热至70℃并溶解的液,并利用均质混合机(日本特殊电装基卡(TokushuKika)公司)来搅拌并乳化。在乳化结束后,将上述溶液冷却至45℃,并相对于组合物的总重量,添加0、0.01、0.05及1重量百分比的藜芦酸进行分散后冷却至30℃。
表3
成分 | 含量(重量百分比) |
藜芦酸 | 0.01、0.05或1 |
荷荷巴油 | 5.0 |
液体石蜡 | 7.0 |
十六芳基醇 | 2.0 |
聚甘油基-3甲基葡萄糖二硬脂酸酯 | 2.0 |
甘油硬脂酸酯 | 0.5 |
角鲨烷 | 3.0 |
丙二醇 | 4.0 |
甘油 | 5.0 |
三乙醇胺 | 0.3 |
羧基乙烯基聚合物 | 0.3 |
生育酚乙酸酯 | 0.2 |
防腐剂、香料 | 微量 |
纯化水 | 余量 |
合计 | 100 |
8-2.含有藜芦酸的营养霜的皱纹改善效果的测定
以30名21至42岁的健康女性为对象,分别在被检查人的脸上使用含有0.01、0.05及1重量百分比的通过上述8-1的方法制备的藜芦酸的营养霜,一天两次,使用3个月。对照组使用了将纯化水包含于其中来制备的营养霜,来代替藜芦酸。
通过测定皮肤弹性的变化来评价了皱纹改善效果,在恒定维持24至26℃温度,38至40%湿度的条件下,利用Cutometer SEM 474(Courage+Khazaka,Cologne,德国)来测定了皮肤弹性,以将没有皮肤弹性的情况表示为0,将皮肤弹性好的情况表示为5作为判断基准,来比较了相对值。在表4中示出了其结果。
表4
实验结果表明,使用含有藜芦酸的营养霜的情况比使用未含有藜芦酸的营养霜的情况,皮肤弹性更增加。并且,随着含有藜芦酸的浓度增加,皮肤弹性也随之提高,从而表示具有浓度依赖性的倾向。
实施例9:以口服方式进行给药的皱纹预防效果
为了测定藜芦酸的皱纹预防效果,将6至7周龄的雌性裸鼠(hairless mouse)(Skh:HR-1)分为正常组、正常组、UV/对照组、UV/藜芦酸组,并在实验期间进行了饲养,每组有8只。在正常组和UV/对照组中,将0.5ml的生理盐水以口服的方式进行给药,在UV/藜芦酸组中,以固体成分为基准,针对每1kg重量,将100mg的藜芦酸和0.5ml的盐水以口服的方式进行给药。
在每周5天的相同时间将制备的上述样品进行给药,在以口服的方式进行给药后,从第2周到第5周,在UV对照组及UV/藜芦酸给药组中,以每周3次,总UV照射量为600mJ/cm2的方式照射了UV。
为了测定皱纹预防效果,在剖检之前,从裸鼠(hairless mouse)的背部利用硅聚合物来采集皮肤模板(replica),同样,在5周后,从各样品处理组中采集皮肤模板来比较了抗老化效果,即,比较了皱纹改善效果。在表5中示出了其结果。
表5
R-参数 | 正常组 | UV/对照组 | UV/藜芦酸 |
R1值 | 0.435±0.0120 | 0.481±0.009 | 0.462±0.013 |
R2值 | 0.357±0.0105 | 0.424±0.010 | 0.405±0.011 |
R3值 | 0.212±0.0097 | 0.237±0.008 | 0.219±0.012 |
R4值 | 0.098±0.0086 | 0.138±0.014 | 0.116±0.010 |
R5值 | 0.231±0.013 | 0.318±0.010 | 0.290±0.013 |
R1值:皮肤的皱纹程度(Skin roughness),R2值:皮肤深处的皱纹程度(Maximum roughness),R3值:皮肤的平均皱纹程度(Average roughness),R4值:皱纹平均深度(Smoothness depth),R5值:皮肤的平均粗糙度(Arithmetic average roughness)(InternationalJournalofCosmeticScience,27(3):155-60(2005))。
实验结果表明,在向裸鼠照射UV的情况下,表示皱纹改善程度的R1、R2、R3、R4及R5值全部增加。观察到将藜芦酸给药的裸鼠(hairless mouse)比未将藜芦酸给药的UV/对照组,减少量更多。
实施例10:藜芦酸对人体皮肤的安全性确认实验
10-1.包含藜芦酸的皮肤外用剂的制备
为了确认藜芦酸对人体皮肤是否也安全,在制备以下列表6的成发及含量包含藜芦酸的皮肤外用剂之后,执行了皮肤安全性验证实验。首先,混合纯化水、甘油、丁二醇,并在约70℃温度下溶解后(水相部分),在约70℃温度下,溶解了除了三个上述成分和三甲醇胺之外的其余成分(油相部分)。之后,将上述油相部分添加于水相部分,利用均质混合机(日本特殊电装基卡(Tokushu Kika)公司)来搅拌并第一次乳化后,最终添加了三甲醇胺。之后,除去在如上所述的混合液中所生成的气泡,再冷却至室温,从而制备了皮肤外用剂。
表6
10-2.皮肤累积刺激性实验
以30名健康成人为对象,对小臂部位,利用通过上述10-1的方法制备的皮肤外用剂,以隔日共9次的方式施行斑贴试验24小时,从而测定了藜芦酸是否刺激皮肤。对照组使用了作为角鲨烷(squalane)基体未含有藜芦酸的皮肤外用剂。
斑贴试验利用Finn小室((Finn chamber),Epitest Ltd,芬兰)来执行,向小室分别滴入15μl的各皮肤外用剂后,实施斑贴试验。利用以下实验式1来将每次在皮肤中出现的反应程度分数化,并在表7中示出了其结果。
实验式1
平均反应度=[[反应指数×反应度/总被检查人数×最高分数(4分)]×100]/检查次数
此时,在反应度中,“±”赋予1分,“+”赋予2分,“++”赋予4分,当平均反应度小于3时,判断为安全的组合物。
表7
实验结果表明,在对照组中,相当于“±”、“+”、“++”的人数在第一次斑贴试验中分别为1名、0名、0名,从第二次斑贴试验开始皮肤无反应。在包含0.1重量百分比及0.5重量百分比的藜芦酸的实验组1及实验组2的情况下,实验的所有实验组中,均没出现皮肤反应。在包含1重量百分比的藜芦酸的实验组3的情况下,相当于“±”、“+”、“++”的人数在第一次斑贴试验中分别为1名、0名、0名,在第二次斑贴试验之后,皮肤无反应。按照如上所述的公式进行计算,其结果表明,对照组和实验组3的平均反应度为0.09,实验组1和实验组2的平均反应度为0.00,均为3以下,从而被判断为是安全的组合物。
制备例1:化妆品的制备
1-1.柔软化妆水的制备
如下列表8,按照常规方法制备了包含藜芦酸作为有效成分的柔软化妆水。
表8
1-2.营养化妆水的制备
如下列表9,按照常规方法制备了包含藜芦酸作为有效成分的营养化妆水。
表9
1-3.营养霜的制备
如下列表10,按照常规方法制备了包含藜芦酸作为有效成分的营养霜。
表10
1-4.按摩霜的制备
如下列表11,按照常规方法制备了包含藜芦酸作为有效成分的按摩霜。
表11
1-5.面膜的制备
如下列表12,按照常规方法制备了包含藜芦酸作为有效成分的面膜。
表12
制备例2:药学制剂的制备
2-1.散剂的制备
为了制备包含藜芦酸作为有效成分的散剂,在混合下列表13的成分之后,填充于气密布来制备了散剂。
表13
成分 | 含量(g) |
藜芦酸 | 2 |
乳糖 | 1 |
2-2.片剂的制备
为了制备包含藜芦酸作为有效成分的片剂,在混合下列表14的成分之后,通过以下常规的片剂的制备方法打锭并制备了片剂。
表14
成分 | 含量(mg) |
藜芦酸 | 100 |
玉米淀粉 | 100 |
乳糖 | 100 |
硬脂酸镁 | 2 |
2-3.胶囊剂的制备
为了制备包含藜芦酸作为有效成分的胶囊剂,在混合下列表15的成分之后,通过以下常规的胶囊剂的制备方法,填充于明胶胶囊来制备了胶囊剂。
表15
成分 | 含量(mg) |
藜芦酸 | 100 |
玉米淀粉 | 100 |
乳糖 | 100 |
硬脂酸镁 | 2 |
Claims (7)
1.一种皮肤状态改善用化妆品组合物,其特征在于,包含藜芦酸或化妆学上能够接受的其盐作为有效成分。
2.根据权利要求1所述的皮肤状态改善用化妆品组合物,其特征在于,上述皮肤状态改善用化妆品组合物为选自由溶液、悬浮液、乳浊液、糊剂、凝胶、乳霜、乳液、粉饼、肥皂、含有表面活性剂的洁面霜、油、粉末粉底、乳浊液粉底、蜡粉底及喷剂组成的组中的一种形态。
3.根据权利要求1所述的皮肤状态改善用化妆品组合物,其特征在于,上述皮肤状态的改善为防紫外线、改善皱纹、促进生发及预防脱发。
4.一种皮肤状态改善用药学组合物,其特征在于,包含藜芦酸或药学上能够接受的其盐作为有效成分。
5.根据权利要求4所述的皮肤状态改善用药学组合物,其特征在于,上述皮肤状态的改善为防紫外线、改善皱纹、促进生发及预防脱发。
6.一种皱纹改善及预防用食品组合物,其特征在于,包含藜芦酸或食品学上能够接受的其盐作为有效成分。
7.根据权利要求1至6中任一项所述的组合物,其特征在于,以组合物的总重量为基准,包含0.00001至15.0重量百分比的上述藜芦酸或其盐。
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