CN112912062A - 含有西伯利亚冷杉油的皮肤屏障强化用组合物 - Google Patents
含有西伯利亚冷杉油的皮肤屏障强化用组合物 Download PDFInfo
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- CN112912062A CN112912062A CN201980069156.6A CN201980069156A CN112912062A CN 112912062 A CN112912062 A CN 112912062A CN 201980069156 A CN201980069156 A CN 201980069156A CN 112912062 A CN112912062 A CN 112912062A
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- skin
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Abstract
本发明涉及一种皮肤屏障强化用组合物,作为有效成分含有西伯利亚冷杉(Abies sibirica)油。
Description
技术领域
本申请要求基于2018年10月16日韩国专利申请第10-2018-0123302号的优先权利益,在该韩国专利申请的文献中公开的所有内容作为一部分包含在本说明书中。
本发明涉及一种含有西伯利亚冷杉油的皮肤屏障强化用组合物。
背景技术
皮肤从外部保护个体而起到所谓屏障功能的非常重要的作用。屏障功能是防御化学物质、大气污染物、干燥的环境、紫外线等之类来自外部的各种刺激和阻止体内水分通过皮肤过度流失的保护功能,这种保护功能只有在由角质形成细胞构成的角质层(Stratumcorneum,horney layer)正常形成时才可以维持其功能。这种皮肤在组织学上由表皮(epidermis)、真皮(dermis)、皮下脂肪(subcutis)三层构成,其中表皮存在于最外部,从而在皮肤老化方面起到最重要的作用,由此在皮肤美容方面也成为最集中的研究对象。
在表皮中,构成最外层的角质层的成分为角质细胞和皮肤脂质,皮肤脂质负责皮肤的屏障功能,这种皮肤的屏障功能通过调节角质的细胞***和分化,保护皮肤免受外部有害物质的影响,防止体内物质的流失且防止皮肤水分蒸发,可以看作是重要的功能。
在皮肤脂质中,负责皮肤屏障功能的主要脂质为表皮的角朊细胞(keratinocytes)分化产生的脂质,该脂质填充分化的角质细胞(corneocytes)之间的空间,也起到赋予细胞之间结合力的作用。这种脂质主要由神经酰胺、胆固醇以及游离脂肪酸构成,其分别占脂质层脂质整体的约40%~65%、10%以及25%。另外,具有一起存在少量的植物鞘氨醇(phytosphingosine)、鞘氨醇(sphingosine)的组成。
另一方面,公知鼻子中存在的嗅觉感受器也包括各种脏器在内的皮肤中。此时,与鼻子不同,公知脏器或者皮肤中存在的嗅觉感受器的功能与在鼻子中时不同。在所述嗅觉感受器中,当OR6M1(嗅觉感受器家族6亚家族M成员1;Olfactory receptor family6subfamily M member 1)在表皮细胞中表达时,可以作为诊断或者评价皮肤屏障功能的生物标志物使用。然而,目前为止尚未公开有与所述嗅觉感受器发生反应的气味,尚未有报告指出利用与所述嗅觉感受器发生反应的皮肤屏障强化功能或者保湿效果功能。
[现有技术文献]
[专利文献]
大韩民国授权专利第10-1023503号
发明内容
为此,本发明人确认到西伯利亚冷杉(Abies sibirica)油与能够诊断或者评价皮肤屏障功能的作为嗅觉感受器的OR6M1发生反应,从而具有强化皮肤屏障或者修复受损的皮肤屏障的效果,由此完成了本发明。
因此,本发明的目的在于提供一种作为有效成分含有西伯利亚冷杉油的皮肤屏障强化用组合物。
为了达成所述目的,本发明提供一种皮肤屏障强化用组合物,作为有效成分含有西伯利亚冷杉(Abies sibirica)油。
本发明的组合物具有强化皮肤屏障的效果。
附图说明
图1是在HEK293细胞中观察OR6M1的细胞膜表达的相片。
图2是基于DMSO处理测定cAMP(环磷酸腺苷)浓度的图表。
图3是基于西伯利亚冷杉油的浓度测定cAMP浓度的图表。
图4是测定因实验例3的紫外线而受损的皮肤屏障修复程度的图表。
具体实施方式
以下,进一步详细说明本发明。
本发明涉及一种皮肤屏障强化用组合物,作为有效成分含有西伯利亚冷杉(Abiessibirica)油。
西伯利亚冷杉(Abies sibirica)油作为从西伯利亚冷杉中提取的油,可以是从西伯利亚冷杉的叶、根、茎、果实或者其混合物中提取的油,但不限于此。
对于所述西伯利亚冷杉油的提取,只要是本领域中公知的方法,则不受特别限定,作为一实现例,可以利用水蒸气蒸馏法(steam distillation)获得。所述水蒸气蒸馏法意指,通过水蒸气进行蒸馏而收集有效成分的方法。
具体为,若将西伯利亚冷杉放入一个设计成允许水蒸气通过的容器中并使其通过,则西伯利亚冷杉的细胞壁被破坏而使细胞壁之间的精华和水蒸气聚集在一起。这些通过管道的同时被冷却而蒸汽被分离为蒸馏液(水溶液成分),精华被分离为油(脂溶性成分),所述油轻而存在于上层部,因此可以通过分离所述上层部,获得西伯利亚冷杉油。
另外,所述西伯利亚冷杉油具有约34种主要成分,其中20重量%至30重量%的莰烯(Camphene)、10重量%至20重量%的δ-3-蒈烯(delta-3-Carene)、10重量%至20重量%的α-蒎烯(alpha-Pinene)、1重量%至5重量%的月桂烯(Myrcene)构成主要成分。
所述西伯利亚冷杉油的特征为,具有松树(pine)香,与皮肤中存在的作为嗅觉感受器的人的OR6M1基因发生反应。
所述OR6M1(嗅觉感受器家族6亚家族M成员1;Olfactory receptor family6subfamily M member 1)基因作为嗅觉感受器,编码嗅细胞的嗅觉感受器6M1(olfactoryreceptor 6M1)蛋白质,OR6M1基因的美国国家生物技术信息中心编号(NCBI accessionID)为NM_001005325.1。
另外,当作为嗅觉感受器的所述OR6M1基因在表皮细胞中表达时,可以作为诊断或者评价皮肤屏障功能的生物标志物使用。
本发明的西伯利亚冷杉油与表皮细胞中存在的嗅觉感受器OR6M1发生反应,可以通过所述反应强化皮肤屏障。
更加具体为,所述西伯利亚冷杉油的气味与表皮细胞中存在的嗅觉感受器OR6M1发生反应,可以通过所述反应强化皮肤屏障。
在本发明中,强化皮肤屏障效果意指,还包括修复受损的皮肤屏障的效果。
因此,本发明的组合物可以表现出强化皮肤屏障以及修复受损的皮肤屏障的效果。
所述皮肤屏障(角质层(Stratum corneum),Skin barrier)由坏死的角质细胞(Coneocyte)和细胞间脂质(Intercellular lipid)构成,作为保护皮肤免受外部刺激的影响,并阻止水分从皮肤蒸发的皮肤保护膜,在皮肤健康中起到核心功能。
因此,本发明的组合物可以还表现出增进皮肤保湿的效果。
本发明的西伯利亚冷杉油可以相对于组合物总重量,含有0.01重量%至10重量%,优选含有0.01重量%至1重量%。
当所述西伯利亚冷杉油含有0.01重量%至1重量%时,不仅适合显示本发明所意图的效果,而且可以将组合物的稳定性以及安全性全部满足,在成本效益方面也是优选的。
本发明的皮肤屏障强化用组合物可以为化妆料组合物。
所述化妆料组合物可以提供为适合局部应用的所有剂型。例如,可以提供为在液相、水相中分散油相得到的乳剂、在油相中分散水相得到的乳剂、悬浮液、固体、凝胶、粉末、糊剂、泡沫(foam)或者气溶胶组合物的剂型。这种剂型的组合物可以根据本领域的常规方法进行制备。
可以是,所述化妆料组合物除了所述物质之外,在不破坏皮肤屏障强化效果的范围内,具体含有能够对皮肤屏障强化效果产生协同效应的其他成分。根据本发明的化妆料组合物可以包括选自由维生素、高分子肽、分子多糖以及神经鞘脂组成的组中的物质。另外,根据本发明的化妆料组合物可以包括保湿剂、润滑剂、表面活性剂、紫外线吸收剂、防腐剂、杀菌剂、抗氧化剂、pH调节剂、有机和无机颜料、香料、凉感剂或者止汗剂。本领域的技术人员可以在不损坏本发明的目的以及效果的范围内,容易地选定所述成分的调配量。
另外,本发明的皮肤屏障强化用组合物可以为药学组合物。
所述药学组合物可以添加将所述组合物作为有效成分而常用的无机或者有机载体,以固体、半固体或者液状形态制成口服给药或者非口服给药制剂。
作为用于所述口服给药的制剂,可以列举出片剂、丸剂、颗粒剂、软、硬胶囊剂、散剂、细粒剂、粉剂、乳剂、糖浆剂、微丸剂等。另外,作为用于所述非口服给药的制剂可以列举出注射剂、点滴剂、软膏、乳液、喷雾、悬浮剂、油剂、栓剂等。为了将本发明的有效成分制剂化,只要按照常规方法就能够容易地进行制剂化,并且可以适当使用表面活性剂、赋形剂、着色剂、香辛料、保存剂、稳定剂、缓冲剂、悬浮剂、其他常用的辅助剂。
根据本发明的药学组合物,其强化皮肤屏障的效果优异,因此可以有效用于治疗以及预防因皮肤屏障受损而引发的皮肤疾病。因所述皮肤屏障受损而引发的皮肤疾病包括特应性皮炎(atopic dermatitis)、干皮症(xeroderma)、牛皮廯(psoriasis)、鱼鳞藓(ichthyosis)、痤疮等,但不限于此。
所述药学组合物可以通过口服、非口服、直肠、局部、经皮、静脉内、肌肉内、腹腔内、皮下等给药。
另外,所述活性成分的给药量将根据待治疗对象的年龄、性别、体重和待治疗的特定疾病或者病理状态、疾病或者病理状态的严重程度、给药途径以及处方者的判断而不同。基于这些因素,所述给药剂量是在本领域技术人员的水平范围内作出的决定。一般给药量为0.001mg/kg/日至2000mg/kg/日,具体为0.5mg/kg/日至1500mg/kg/日。
另外,本发明的皮肤屏障强化用组合物可以为食品组合物。
根据本发明的食品组合物除了上述成分之外,可以还以不阻碍西伯利亚冷杉油的功效的水平的量,进一步包括具有强化皮肤屏障效果的其他成分。例如,可以包括糖分、酸、糖醇中的任一个以上。
根据本发明的食品组合物可以为健康食品、功能性食品以及食品添加剂组合物。所述组合物可以通过包括添加各种赋形剂或者添加剂的步骤的常规方法,应用为片剂、丸剂、胶囊剂、颗粒剂、饮品剂、焦糖、减脂棒、茶包等各种剂型。除了有效成分之外,可以根据剂型或者使用目的,本领域技术人员毫不费力地适当选择和调配本领取中通常使用的成分,并将其调配到组合物中,当与其他成分调配时,可以产生协同效应。
用于实施发明的方式
以下,为了具体说明本发明,列举实施例进行详细说明。然而,根据本发明的实施例可以变形为各种不同方式,本发明的范围不应被解释为限于以下详细叙述的实施例。本发明的实施例是为了向本领域技术人员更加完整地说明本发明而提供的。
实施例1.西伯利亚冷杉(Abies sibirica)油
作为西伯利亚冷杉油,使用了Roberter公司的产品。
比较例1.樟子松(Pinus sylvestris)油
使用了樟子松油。
实验例1.OR6M1基因的细胞膜表达的观察
为了使作为嗅觉感受器的OR6M1基因过表达,使用了韩国大邱庆北科学技术院制造的向pcDNA(质粒克隆DNA;plasmid cloning DNA)加入OR6M1基因的质粒(plasmid)。
将含所述OR6M1基因的质粒注入到不含OR6M1基因的HEK293细胞中,通过免疫细胞化学法(immunocytochemistry)确认了OR6M1在所述HEK293细胞中是否正常表达(图1)。
此时,作为第一抗体(Primary antibody)使用了anti-FLAG(mouse(小鼠),Sigma#M1804),作为第二抗体(Secondary antibody)使用了anti-mouse(Alexa 568,Abcam#ab175472),为了成像,使用了共聚焦显微镜(Confocal microscope,LSM700,Zeiss,400x)。
在图1的结果中,红色表示作为嗅觉感受器的OR6M1,可以确认到所述OR6M1在HEK293细胞中过表达。
实验例2.西伯利亚冷杉油与OR6M1基因反应性的确认
为了使作为嗅觉感受器的OR6M1基因过表达,使用了韩国大邱庆北科学技术院制造的向pcDNA(plasmid cloning DNA)加入OR6M1基因的质粒(plasmid)。
将含所述OR6M1基因的质粒注入到不含OR6M1基因的HEK293细胞中。
另外,为了使作为嗅觉感受器的OR6V1基因过表达,使用了韩国大邱庆北科学技术院制造的向pcDNA(plasmid cloning DNA)加入OR6V1基因的质粒(plasmid)。
将含所述OR6V1基因的质粒注入到不含OR6V1基因的HEK293细胞中。
当利用DMSO溶剂处理所述注入有OR6M1基因的HEK293细胞以及注入有OR6V1基因的HEK293细胞时,未见注入有所述OR6M1以及OR6V1基因的各个HEK293细胞的cAMP浓度发生变化(图2)。
将在所述实施例1中制备的西伯利亚冷杉油溶解到DMSO溶剂中,制备0.1ppm(10-5%)、1ppm(10-4%)、10ppm(10-3%)以及100ppm(10-2%)溶液。
利用所述浓度的西伯利亚冷杉油分别处理所述注入有OR6M1基因的HEK293细胞以及注入有OR6V1基因的HEK293细胞,观察HEK293细胞的cAMP浓度(图3)。
其结果,OR6V1基因不与西伯利亚冷杉油发生反应,OR6M1基因与西伯利亚冷杉油发生反应而HEK293细胞的cAMP浓度增加,显示出依赖于西伯利亚冷杉油浓度的结果。
因此,可以确认到作为嗅觉感受器的OR6M1基因与西伯利亚冷杉油选择性地发生反应。
实验例3.受损的皮肤屏障的修复效果的测定
测定了西伯利亚冷杉油对因紫外线导致的皮肤损伤而受损的皮肤屏障功能的修复效果。
将新生儿的角质形成细胞(normal human epidermal keratinocyte:P988,Lonza)使用角质形成培养基(KGM)在60mm的细胞培养皿(cell culture dish)中以1.25×104cells/dish(细胞/皿)密度进行分装之后,在37℃、5%CO2培养器中培养至融合(confluency)80%左右。
将所述实施例1的西伯利亚冷杉油溶解到DMSO溶剂中,制备100ppm(10-2%)溶液。
另外,将所述比较例1的樟子松(Pinus sylvestris)油溶解到DMSO溶剂中,制备100ppm(10-2%)溶液。
然后,将对所述角质形成细胞进行紫外线(UVB 25Mj/cm2)处理、一起进行紫外线(UVB 25Mj/cm2)以及100ppm的西伯利亚冷杉油溶液处理、一起进行紫外线(UVB 25Mj/cm2)以及100ppm的樟子松油溶液处理的各个细胞培养2天之后,确认了作为表皮分化标志物的角蛋白(keratin)-1(KRT1,Hs01549614_g1)以及角蛋白(keratin)-10(KRT10,Hs01043114_g1)的基因变化。
各个表皮分化标志物表达变化是通过去除细胞生长培养基,添加1mL的试剂(Trizol,Invitrogen),按照英杰(invitrogen)公司的RNA分离法分离RNA之后,利用紫外线检测器(HEWLETT PACKARD)在260nm下定量RNA之后,进行逆转录-聚合酶链反应(Reversetranscription-polymerase chain reaction;RT-PCR)。为了各样本对KRT1和KRT10的基因分析,使用了探针法(taqman probe;Hs01549615_g1,Hs00166289_m1),以作为互补基因的RPL13A(Hs01578912_m1)为基准进行了校正。
其结果,当将未处理紫外线的角质形成细胞的KRT1的mRNA表达量(con)作为基准时,可以确认到仅进行紫外线处理的KRT1的mRNA表达量显著减少。然而,一起进行紫外线以及西伯利亚冷杉油处理的KRT1的mRNA表达量相比仅进行紫外线处理的结果,显示高的表达量,从所述结果可以确认到西伯利亚冷杉油具有修复受损的皮肤屏障的效果。虽然一起进行紫外线以及樟子松油处理的KRT1的mRNA表达量略高于仅进行紫外线处理的结果,但远低于一起进行紫外线以及西伯利亚冷杉油处理的结果。另外,KRT10的测定结果也与KRT1类似(图4)。
从所述实验例1至实验例3的结果可以可知,由于角质形成细胞具有作为嗅觉感受器的OR6M1,因此西伯利亚冷杉油与细胞的OR6M1发生反应而修复受损的皮肤屏障,所述樟子松油不与OR6M1发生反应而无法修复受损的皮肤屏障。
因此,可知西伯利亚冷杉油具有强化皮肤屏障的效果。
剂型例1.营养化妆水
按照下表1中记载的组分,通过常规方法制备营养化妆水。
[表1]
剂型例2.营养乳液
按照下表2中记载的组分,通过常规方法制备营养乳液。
[表2]
原料名 | 含量(重量%) |
净化水 | 剩余量 |
甘油 | 3 |
丁二醇 | 3 |
液体石蜡 | 5 |
β-葡聚糖 | 7 |
实施例1的西伯利亚冷杉油 | 1 |
卡波姆 | 0.1 |
辛酸/癸酸甘油三酯 | 3 |
角鲨烷 | 5 |
鲸蜡硬脂基葡糖苷 | 1.5 |
山梨坦硬脂酸酯 | 0.4 |
吐温60 | 1.5 |
防腐剂 | 适量 |
香料 | 适量 |
色素 | 适量 |
三乙醇胺 | 0.1 |
计 | 100 |
剂型例3.营养霜
按照下表3中记载的组分,通过常规方法制备营养霜。
[表3]
原料名 | 含量(重量%) |
甘油 | 3.5 |
液体石蜡 | 3 |
β-葡聚糖 | 7 |
卡波姆 | 7 |
实施例1的西伯利亚冷杉油 | 1 |
辛酸/癸酸甘油三酯 | 1 |
角鲨烷 | 3 |
鲸蜡硬脂基葡糖苷 | 5 |
山梨坦硬脂酸酯 | 1.5 |
吐温60 | 0.4 |
三乙醇胺 | 1.2 |
防腐剂、香料 | 适量 |
净化水 | 剩余量 |
计 | 100 |
剂型例4.局部给药用药剂(贴片剂)的制备
按照下表4中记载的组分,通过常规方法制备局部给药用药剂(贴片剂)。
[表4]
原料名 | 含量(重量%) |
实施例1的西伯利亚冷杉油 | 1 |
β-1,3-葡聚糖 | 3 |
二乙胺 | 0.7 |
亚硫酸钠 | 0.1 |
聚氧乙烯月桂醚(E.O=9) | 1 |
聚羟基乙烯十六烷基硬脂醚(Cetomacrogol 1000) | 1 |
粘性的石蜡油 | 2.5 |
辛酸酯/癸酸酯(Cetiol LC) | 2.5 |
聚乙二醇400 | 3 |
聚丙烯酸(Carbopol 934P) | 1 |
净化水 | 剩余量 |
计 | 100 |
剂型例5.片剂的制备
混合2mg的所述实施例1的西伯利亚冷杉油、100mg的玉米淀粉、100mg的乳糖以及2mg的硬脂酸镁之后,按照常规的片剂的制备方法进行压片制得。
剂型例6.丸的制备
混合0.03g的所述实施例1的西伯利亚冷杉油、1.5g的乳糖、1g的甘油以及0.5g的木糖醇之后,按照常规的制备方法制成每丸为4g。
剂型例7.饮品剂的制备
混合360mg的所述实施例1的西伯利亚冷杉油、10g的葡萄糖、0.6g的柠檬酸以及25g的液状低聚糖之后,添加300mL的净化水,向每瓶填充200mL。填充到瓶中之后,在130℃下杀菌4~5秒,制备饮料。
剂型例8.焦糖剂型的制备
混合58mg的所述实施例1的西伯利亚冷杉油、1.8g的玉米糖浆(corn syrup)、0.5g的脱脂牛奶、0.5g的大豆卵磷脂、0.6g的黄油、0.4g的植物性硬化油、1.4g的白糖、0.58g的人造黄油以及20mg的食盐,制备焦糖剂型。
Claims (8)
1.一种皮肤屏障强化用组合物,作为有效成分含有西伯利亚冷杉(Abies sibirica)油。
2.根据权利要求1所述的皮肤屏障强化用组合物,其特征在于,
所述西伯利亚冷杉油与皮肤中存在的作为嗅觉感受器的OR6M1基因发生反应。
3.根据权利要求1所述的皮肤屏障强化用组合物,其特征在于,
所述西伯利亚冷杉油为西伯利亚冷杉的叶、根、茎、果实或者它们的混合物的油。
4.根据权利要求1所述的皮肤屏障强化用组合物,其特征在于,
所述西伯利亚冷杉油相对于组合物总重量,含有0.01重量%至10重量%。
5.根据权利要求1所述的皮肤屏障强化用组合物,其特征在于,
所述组合物具有增进皮肤保湿效果。
6.根据权利要求1所述的皮肤屏障强化用组合物,其特征在于,
所述组合物为化妆料组合物。
7.根据权利要求1所述的皮肤屏障强化用组合物,其特征在于,
所述组合物为药学组合物。
8.根据权利要求1所述的皮肤屏障强化用组合物,其特征在于,
所述组合物为食品组合物。
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