CN116348102A - 包含异原莪述烯醇的用于防老化或皮肤再生的组合物 - Google Patents
包含异原莪述烯醇的用于防老化或皮肤再生的组合物 Download PDFInfo
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- CN116348102A CN116348102A CN202180052690.3A CN202180052690A CN116348102A CN 116348102 A CN116348102 A CN 116348102A CN 202180052690 A CN202180052690 A CN 202180052690A CN 116348102 A CN116348102 A CN 116348102A
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Abstract
Description
技术领域
本发明涉及用于防老化或皮肤再生的组合物,并且更特别地,涉及包含异原莪述烯醇(Isoprocurcumenol)的用于防老化或皮肤再生的组合物。
背景技术
皮肤老化是不可避免的现象。老化导致皮肤弹性的损失,产生皱纹并引起诸如色素沉着和表皮萎缩的症状。皮肤老化主要分为生理老化(其指示皮肤的功能、结构和形状的变化)和由外部刺激例如紫外线引起的老化。
随着皮肤的老化,细胞的功能下降,并且同时,皮肤受到外部环境例如紫外线的影响。首先,随着老化的进行,角质形成细胞的分裂速率减慢。虽然正常的皮肤再生周期为约28天,但这个周期随着老化而增大。其次,已知存在于表皮层中的成纤维细胞的数量随着老化而减少,并且同时,功能下降。成纤维细胞是产生胶原蛋白和弹性蛋白纤维以为皮肤提供弹性的细胞。然而,当成纤维细胞的数量减少并且其功能下降时,发生真皮萎缩。最后,基质蛋白酶的表达随着老化而增加。因此,胶原蛋白和弹性蛋白纤维被基质蛋白酶分解,从而导致真皮萎缩的增加。
然而,即使在100岁年龄的生活中不能完全防止老化,对可以改善皮肤老化以增强美感的物质的期望和需求也在增加。因此,需要快速开发减缓或改善皮肤老化的物质。
表皮生长因子(Epidermal growth factor,EGF)是调控表皮或表皮细胞生长和存活的生长因子。EGF也被认为是与癌症相关的因子,并且已在医学上用于治疗伤口和烧伤。近来,EGF已被广泛用作用于化妆品的原料,并且已经对包含EGF的用于防止皮肤老化的化妆品组合物进行了许多研究,例如韩国专利第2093884号。因此,需要研究具有与EGF的功能类似的功能并且稳定且可用于通过皮肤屏障的天然来源的小分子物质。然而,已知EGF的生物合成、分离和纯化是昂贵的,并且难以以活性状态储存EGF。此外,存在EGF由于其分子量大而不能容易地通过皮肤屏障的问题。
因此,需要研究具有与EGF的功能类似的功能并且稳定且可用于通过皮肤屏障的天然来源的小分子物质。
发明内容
技术问题
本公开内容的一个方面是提供用于防老化或皮肤再生的组合物,所述组合物包含异原莪述烯醇(其为一种天然来源的小分子物质)或其盐作为活性成分以提供具有与EGF的功能类似的功能并且稳定且可用于通过皮肤屏障的用于防老化或皮肤再生的组合物。
技术方案
根据本公开内容的一个方面,本发明提供了用于防老化或皮肤再生的组合物,其包含作为活性成分的由下式(1)表示的异原莪述烯醇或其盐:
[式(1)]
有益效果
根据本公开内容的一个方面,异原莪述烯醇可以从各种植物物种例如莪术(zedoaria)中提取。根据本发明的组合物具有通过使用其而不具有副作用的优点。此外,与EGF一样,异原莪述烯醇与EGFR结合以激活与细胞的存活、生长和增殖相关的信号,从而增加皮肤细胞对外部刺激的抗性和生存力,并且增强由于外部刺激或老化而受损的皮肤再生。此外,虽然异原莪述烯醇具有与EGF的功能类似的功能,但与EGF不同的是,异原莪述烯醇稳定且尺寸小,并因此具有能够移动通过皮肤屏障到达皮肤基底层以执行功能的优点。因此,本发明的异原莪述烯醇可以以多种方式使用,例如在药物、准药物、化妆品、香料材料、功能性生物材料和功能性食品材料中使用。
附图说明
图1示出了确定在通过细胞内注射EGFR活性测量生物传感器系统使生物传感器蛋白(EGFP-SH2)表达时,该蛋白质通过EGF以及还通过异原莪述烯醇表达的结果。
图2示出了确定取决于异原莪述烯醇浓度的对作为角质形成细胞的HaCaT细胞的细胞毒性的结果。
图3示出了确定作为EGFR亚型的ERK和AKT被异原莪述烯醇激活的结果。
图4示出了确定通过异原莪述烯醇增加的egr1、c-myc、c-jun和c-fos基因的表达模式的结果。
图5示出了通过CCK-8测定确定通过异原莪述烯醇的角质形成细胞的生长促进作用的结果。
图6示出了显示出通过伤口愈合评价来确定通过异原莪述烯醇的角质形成细胞的增殖和生长促进作用的结果的照片(a)和图(b)。
图7示出了在由UVB引起的细胞损伤的条件下角质形成细胞的抗性由于异原莪述烯醇而增加。
具体实施方式
在下文中,将参照附图描述本发明的优选实施方案。然而,本发明的实施方案可以被修改成各种形式,并且本发明的范围不限于以下描述的实施方案。
发明人通过在实验上确定作为天然来源的小分子物质的异原莪述烯醇具有与表皮生长因子(EGF)的功能类似的功能并且稳定且尺寸小从而易于移动通过皮肤屏障而完成了本发明。
本发明涉及由下式(1)表示的异原莪述烯醇或其盐用于制备用于防老化或皮肤再生的组合物的用途:
[式(1)]
具体地,本发明提供了用于防老化或皮肤再生的组合物,其包含作为活性成分的由下式(1)表示的异原莪述烯醇或其盐:
[式(1)]
异原莪述烯醇可以从各种植物物种例如莪术的提取物中获得,并且具有与细胞中激活与细胞的存活、生长和增殖相关的信号的表皮生长因子(EGF)的功能类似的功能,从而增加皮肤细胞对外部刺激的抗性和生存力,并且增强由于外部刺激或老化而受损的皮肤再生。
本发明的组合物可以以10nM至100μM、或1μM至100μM,例如5μM至100μM,优选10μM至100μM,并且更优选25μM至50μM的量包含异原莪述烯醇或其盐。当异原莪述烯醇或其盐的浓度小于1μM时,异原莪述烯醇或其盐的作用可能无法充分表达,当浓度大于100μM时,细胞毒性可能显著增大。
本发明的“皮肤再生”可以是指促进皮肤细胞的生长并且增强生存力以防止来自外部刺激或内部刺激的细胞损伤,减少皮肤皱纹和增强皮肤弹性。此外,皮肤再生包括改善或治疗由外部环境引起的皮肤损伤,例如由烧伤引起的皮肤和组织损伤的含义。由此,本发明的组合物可以改善或治疗烧伤伤口。
此外,本发明的异原莪述烯醇或其盐可以与EGFR结合以促进细胞增殖或生长,从而激活防老化或皮肤再生。通过本发明的异原莪述烯醇或其盐增殖或生长的细胞可以为表皮细胞,优选为角质形成细胞(其为产生角蛋白的细胞),但本发明不限于此。
作为构成动物内各种组织的主要成分的蛋白质的角蛋白使毛发有光泽和弹性,并赋予皮肤生命力和弹性。此外,皮肤的角质层中的角蛋白蛋白质对化学品具有强的抗性,从而针对对于皮肤的化学刺激执行保护功能。
因此,本发明的组合物可以增强皮肤弹性或改善皮肤皱纹的问题。
同时,本发明的组合物可以增强对由于外部刺激而导致的细胞损伤的抗性,从而防止或改善由于外部刺激而导致的细胞损伤。外部刺激可以为由化妆品或其他外用剂导致的化学刺激、物理刺激或紫外线,优选紫外线,但本发明不限于此。
此外,本发明的组合物可以为肠胃外组合物或经口组合物,例如药物组合物、化妆品组合物、或健康功能性食品组合物。
具体地,当本发明的组合物为药物组合物时,所述组合物可以另外地包含可药用载体,并且所述组合物可以另外地包含在制备药物组合物时通常使用的合适的载体、模板和稀释剂。组合物还可以根据本领域常规已知的方法以经口制剂、外用制剂、栓剂(例如散剂、颗粒剂、片剂、胶囊剂、混悬剂、乳剂、糖浆剂和气雾剂)和无菌注射液的形式进行配制和使用。
根据本发明的药物组合物可以以外用制剂(例如散剂、颗粒剂、片剂、胶囊剂、混悬剂、乳剂、糖浆剂和气雾剂)和无菌注射液的形式进行配制和使用,并且可以优选具有乳膏剂、凝胶剂、贴剂、喷雾剂、软膏剂、硬膏剂、洗剂、搽剂、糊剂或敷剂的制剂。可以包含在所述组合物中的载体、赋形剂和稀释剂可以为包含以下中的至少一者的化合物:乳糖、右旋糖、蔗糖、寡糖、山梨糖醇、甘露糖醇、木糖醇、赤藓糖醇、麦芽糖醇、淀粉、阿拉伯胶、藻酸盐、明胶、磷酸钙、硅酸钙、纤维素、甲基纤维素、微晶纤维素、聚乙烯吡咯烷酮、水、羟基苯甲酸甲酯、羟基苯甲酸丙酯、滑石、硬脂酸镁和矿物油。
所述组合物可以使用稀释剂或赋形剂(例如,填料、增量剂、粘结剂、润湿剂、崩解剂和表面活性剂)来制备。例如,用于经口施用的固体包括片剂、丸剂、散剂、颗粒剂、胶囊剂等,并且这样的固体可以通过将提取物与至少一种赋形剂(例如淀粉、碳酸钙、蔗糖、乳糖和明胶)混合来制备。除了简单的赋形剂之外,还可以使用润滑剂,例如硬脂酸镁滑石。除了本领域通常使用的简单稀释剂例如水和液体石蜡之外,用于经口使用的液体制剂(例如混悬剂、溶液剂、乳剂和糖浆剂)还可以包含各种赋形剂(例如润湿剂、甜味剂、空气清新剂和防腐剂)。
用于肠胃外施用的制剂可以包括无菌水溶液、非水溶剂、混悬剂、乳剂、冻干剂和栓剂。非水溶剂和混悬剂可以包括丙二醇、聚乙二醇、植物油例如橄榄油、和可注射酯例如油酸乙酯。可以使用Witepsol、聚乙二醇、Tween 61、可可豆脂、月桂精、甘油明胶等作为用于栓剂的基质。
根据本发明的药物组合物可以以药理学有效量施用。在本发明中,“药理学有效量”是指足以以适用于医学治疗的合理的效益/风险比来治疗疾病的量。有效剂量水平可以根据患者疾病的类型和严重程度、药物的活性、对药物的敏感性、施用的持续时间和途径以及排泄的速率、治疗的时间段、诸如同时使用的药物的因素,以及医学领域公知的其他因素来确定。根据本发明的药物组合物可以作为单一治疗剂施用或与其他治疗剂组合施用,可以与常规治疗剂顺序施用或同时施用,并且可以单次施用或多次施用。鉴于所有以上因素,重要的是施用可以获得最大效果而不引起副作用的最小量。这样的量可以容易地由本发明所属领域的技术人员确定。
本发明的药物组合物可以通过多种途径施用于对象。本发明的药物组合物根据各种因素(例如,要治疗的疾病,施用的途径,患者的年龄、性别和体重,以及疾病的严重程度)和作为活性成分的药物的类型来确定。
本发明的组合物可以以健康功能性食品组合物的形式提供。此外,为了皮肤再生的目的(例如,防老化、增强皮肤弹性和改善皮肤皱纹的问题),可以将所述组合物添加到食品中。根据本发明的健康功能性食品组合物可以按照原样使用或者与其他食品或食品成分一起使用,并且可以根据常规方法适当地使用。待混合的活性成分的量可以根据预期用途例如预防和保健或治疗性治疗适当地确定。当制备食品或饮料时,基于总组合物的重量,异原莪述烯醇或其盐通常可以以大于0重量%且多至15重量%,优选1重量%至10重量%或更小的量添加到根据本发明的健康功能性食品组合物中。然而,异原莪述烯醇在稳定性方面没有问题,因此可以以等于或大于以上范围的量作为活性成分使用。
食品的类型没有特别限制,但可以包含所述材料的食品的实例为肉类、香肠、面包、巧克力、糖果、小吃、糖果点心、比萨、拉面、其他面条、口香糖、和乳制品(包括冰淇淋)、各种汤、饮料、茶、饮品、维生素复合物等,并且可以包括常规意义上的所有健康功能性食品。
与常规饮料一样,该饮料可以包含各种调味剂或天然碳水化合物作为另外的成分。天然碳水化合物为单糖例如葡萄糖和果糖,二糖例如麦芽糖和蔗糖,多糖例如糊精和环糊精,以及糖醇例如木糖醇、山梨糖醇和赤藓糖醇。甜味剂可以包括天然甜味剂例如奇异果甜蛋白和甜叶菊提取物、或合成甜味剂例如糖精和阿斯巴甜。天然碳水化合物的量可以为每100mL的根据本发明的健康功能性食品组合物约0.01g至0.20g,优选约0.04g至0.10g。
此外,根据本发明的健康功能性食品组合物可以包含各种营养补充剂、维生素、电解质、调味剂、着色剂、果胶酸及其盐、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、用于碳酸饮料的碳酸化剂等。此外,根据本发明的健康功能性食品组合物可以包含用于制备天然果汁、果汁饮料和蔬菜饮料的果肉。这些组分可以单独使用或组合使用。这些添加剂的量不是非常重要,但是可以在每100重量份的根据本发明的健康功能性食品组合物0.01重量份至0.20重量份的范围内选择。
本发明的组合物可以以化妆品组合物的形式提供。根据本发明的化妆品组合物的制剂可以呈以下形式:皮肤洗剂、皮肤软化剂、紧肤剂、收敛剂、洗剂、乳液洗剂、保湿乳液、滋养乳液、按摩霜、滋养霜、保湿霜、护手霜、粉底、精华液、滋养精华液、剥撕式面膜、皂、清洁泡沫、清洁洗剂、清洁霜、身体洗剂或身体清洁剂。
根据本发明的化妆品组合物可以另外地包含选自水溶性维生素、油溶性维生素、高分子肽、高分子多糖和鞘脂的化合物。
可以使用任何水溶性维生素,只要其可以添加到化妆品组合物中即可。优选地,可以使用维生素B1、维生素B2、维生素B6、吡哆素、吡哆素盐酸盐、维生素B12、泛酸、烟酸、烟酰胺、叶酸、维生素C、维生素H等。还可以使用其盐(例如,盐酸硫胺素和抗坏血酸钠盐)或其衍生物(例如,抗坏血酸-2-磷酸钠盐和抗坏血酸-2-磷酸镁盐)作为本发明中的水溶性维生素。水溶性维生素可以通过常规方法(例如,微生物转化法、从微生物培养物中纯化的方法和酶促法或化学合成法)获得。
可以使用任何可用的维生素,只要其可以添加到化妆品中即可,并且优选地包括维生素A、胡萝卜素、维生素D2、维生素D3、维生素E(例如,d1-α生育酚、d-α生育酚和d-α生育酚)等。还可以使用其衍生物(例如,抗坏血酸棕榈酸酯、抗坏血酸硬脂酸酯、抗坏血酸二棕榈酸酯、dl-α生育酚乙酸酯、dl-α生育酚烟酸酯、维生素E、DL-泛醇、D-泛醇和泛酰基乙醚)作为本发明中的油溶性维生素。可用的维生素可以通过常规方法(例如,微生物转化法、从微生物培养物中纯化的方法和酶促法或化学合成法)获得。
可以使用任何高分子肽,只要其可以添加到化妆品中即可,并且优选地包括胶原蛋白、水解胶原蛋白、明胶、弹性蛋白、水解弹性蛋白、角蛋白等。高分子肽可以通过常规方法例如从微生物培养基中纯化的方法、酶促法或化学合成法来获得,或者可以常规地通过从天然产物例如猪或牛的真皮或蚕丝纤维中纯化来获得。
可以使用任何高分子多糖,只要其可以添加到化妆品中即可,并且优选地包括羟乙基纤维素、黄原胶、透明质酸钠、硫酸软骨素或其盐(例如,钠盐)。例如,硫酸软骨素或其盐可以常规地通过从哺乳动物或鱼类中纯化来获得。
可以使用任何鞘脂,只要其添加到化妆品中即可,并且优选地包括神经酰胺、植物鞘氨醇、鞘糖脂等。鞘脂可以通过常规方法从哺乳动物、鱼类、贝类、酵母菌、或植物中纯化,或者可以通过化学合成法来获得。
除了上述必需成分之外,本发明的化妆品组合物还可以根据需要包含通常添加到化妆品中的其他成分。
可以添加的其他成分包括油和脂肪组分、保湿剂、润肤剂、表面活性剂、有机或无机颜料、有机粉末、紫外线吸收剂、防腐剂、杀菌剂、抗氧化剂、植物提取物、pH调节剂、醇类、颜料、芳香剂、血液循环促进剂、清凉剂、止汗剂、纯净水等。
油和脂肪组分可以包括基于酯的油和脂肪、基于烃的油和脂肪、基于有机硅的油和脂肪、基于氟的油和脂肪、动物油和脂肪、植物油和脂肪等。
酯类油和脂肪可以包括三-2-乙基己酸甘油酯、2-乙基己酸十六烷基酯、肉豆蔻酸异丙酯、肉豆蔻酸丁酯、棕榈酸异丙酯、硬脂酸乙酯、棕榈酸辛酯、异硬脂酸异十六烷基酯、硬脂酸丁酯、亚油酸乙酯、亚油酸异丙酯、油酸乙酯、肉豆蔻酸异十六烷基酯、肉豆蔻酸异硬脂酯、棕榈酸异硬脂酯、肉豆蔻酸辛基十二烷基酯、异硬脂酸异十六烷基酯、癸二酸二乙酯、己二酸二异丙酯、新戊酸异烷基酯、甘油三(辛酸酯或癸酸酯)、三羟甲基丙烷三(2-乙基己酸酯)、三羟甲基丙烷三异硬脂酸酯、季戊四醇四(2-乙基己酸酯)、辛酸十六烷基酯、月桂酸癸酯、月桂酸己酯、肉豆蔻酸癸酯、肉豆蔻酸肉豆蔻酯、肉豆蔻酸十六烷基酯、硬脂酸硬脂酯、油酸癸酯、蓖麻油酸十六烷基酯、月桂酸异硬脂酸酯、肉豆蔻酸异十三烷基酯、棕榈酸异十六烷基酯、硬脂酸辛酯、硬脂酸异十六烷基酯、油酸异癸酯、油酸辛基十二烷基酯、亚油酸辛基十二烷基酯、异硬脂酸异丙酯、2-乙基己酸鲸蜡硬脂酯、2-乙基己酸硬脂酯、异硬脂酸己酯、乙二醇二辛酸酯、乙二醇二油酸酯、丙二醇二癸酸酯、丙二醇二(辛醇或癸酸)、丙二醇二辛酸酯、新戊二醇二癸酸酯、新戊二醇二辛酸酯、三辛酸甘油酯、十三烷酸甘油酯、三异棕榈酸甘油酯、三异硬脂酸甘油酯、新戊酸辛基十二烷基酯、辛酸异硬脂酸酯、异壬酸辛酯、新癸酸己基癸酯、新癸酸辛基十二烷基酯、异硬脂酸异十六烷基酯、异硬脂酸异硬脂酯、异硬脂酸辛基癸酯、聚甘油油酸酯、聚甘油异硬脂酸酯、柠檬酸三异十六烷基酯、柠檬酸三异烷基酯、柠檬酸三异辛酯、乳酸月桂酯、乳酸肉豆蔻酯、乳酸十六烷基酯、乳酸辛基癸酯、柠檬酸三乙酯、柠檬酸乙酰基三乙酯、柠檬酸乙酰基三丁酯、柠檬酸三辛酯、苹果酸二异硬脂酯、羟基硬脂酸2-乙基己酯、琥珀酸二(2-乙基己基)酯、己二酸二异丁酯、癸二酸二异丙酯、癸二酸二辛酯、硬脂酸胆甾醇酯、异硬脂酸胆甾醇酯、羟基硬脂酸胆甾醇酯、油酸胆甾醇酯、油酸二氢胆甾醇酯、异硬脂酸植物甾醇酯、油酸植物甾醇酯、12-硬脂酰羟基硬脂酸异十六烷基酯、12-硬脂酰羟基硬脂酸硬脂酯、12-硬脂酰羟基硬脂酸酯异硬脂酯等。
基于烃的油和脂肪可以包括角鲨烯、液体石蜡、α-烯烃低聚物、异构烷烃、地蜡、石蜡、液体异构烷烃、聚丁烯、微晶蜡、矿脂等。
基于有机硅的油和脂肪可以包括聚甲基硅酮、甲苯基硅酮、甲基环聚硅氧烷、八甲基聚硅氧烷、十甲基聚硅氧烷、十二甲基环硅氧烷、二甲基硅氧烷/甲基十六基氧基硅氧烷共聚物、二甲基硅氧烷/甲基硬脂基氧基硅氧烷共聚物、经烷基改性的硅油、经氨基改性的硅油等。
基于氟的油和脂肪可以包括全氟聚醚等。
动物或植物油和脂肪可以包括鳄梨油、扁桃仁油、橄榄油、麻油、米糠油、红花油、豆油、玉米油、菜籽油、杏仁油、棕榈仁油、棕榈油、蓖麻油、葵籽油、葡萄籽油、棉籽油、椰子油、夏威夷坚果油、小麦胚芽油、米胚芽油、乳木果油、月见草油、澳洲坚果油、白芒花籽油、卵黄油、牛脂、马油、貂油、罗非鱼油、霍霍巴油、小烛树蜡、巴西棕榈蜡、液体羊毛脂、氢化蓖麻油等。
润湿剂可以包括水溶性低分子保湿剂、脂溶性分子保湿剂、水溶性聚合物、或油溶性聚合物。
水溶性低分子保湿剂可以包括丝氨酸、谷氨酰胺、山梨糖醇、甘露糖醇、吡咯烷酮-羧酸钠、甘油、丙二醇、1,3-丁二醇、乙二醇、聚乙二醇B(聚合度n=2或更大)、聚丙二醇(聚合度n=2或更大)、聚甘油B(聚合度n=2或更大)、乳酸、乳酸盐/酯等。
脂溶性低分子保湿剂可以包括胆固醇、胆固醇酯等。
水溶性聚合物可以包括羧基乙烯基聚合物、聚天冬氨酸盐/酯、黄芪胶、黄原胶、甲基纤维素、羟甲基纤维素、羟乙基纤维素、羟丙基纤维素、羧甲基纤维素、水溶性壳聚糖、甲壳糖、糊精等。
脂溶性聚合物可以包括聚乙烯吡咯烷酮/二十烯共聚物、聚乙烯吡咯烷酮/十六碳烯共聚物、硝酸纤维素、糊精脂肪酸酯、高分子有机硅等。
润肤剂可以包括长链酰基谷氨酸胆固醇酯、羟基硬脂酸胆固醇酯、12-羟基硬脂酸酯、硬脂酸、松香酸、羊毛脂脂肪酸胆固醇酯等。
表面活性剂可以包括非离子表面活性剂、阴离子表面活性剂、阳离子表面活性剂、两性表面活性剂等。
非离子表面活性剂可以包括自乳化单硬脂酸甘油酯、丙二醇脂肪酸酯、甘油脂肪酸酯、聚甘油脂肪酸酯、脂肪酸脱水山梨糖醇酯、聚氧乙烯(POE)脱水山梨糖醇脂肪酸酯、POE脱水山梨糖醇脂肪酸酯、POE甘油脂肪酸酯、POE烷基醚、POE脂肪酸酯、POE氢化蓖麻油、POE蓖麻油、POE/POP(聚氧乙烯/聚氧丙烯)共聚物、POE/POP烷基醚、经聚醚改性的有机硅、月桂酸烷醇酰胺、烷基胺氧化物、氢化大豆磷脂等。
阴离子表面活性剂可以包括脂肪酸皂、α-酰基磺酸盐、烷基磺酸盐、烷基烯丙基磺酸盐、烷基萘磺酸盐、烷基硫酸盐、POE烷基醚硫酸盐、烷基酰胺硫酸盐、烷基磷酸盐、POE烷基磷酸盐、烷基酰胺磷酸盐、烷基酰基烷基牛磺酸盐、N-酰基氨基酸盐、POE烷基醚羧酸盐、烷基磺基琥珀酸盐、烷基磺基乙酸钠、酰化水解胶原肽盐、全氟烷基磷酸酯等。
阳离子表面活性剂可以包括氯化烷基三甲基铵、硬脂基三甲基氯化铵、硬脂基三甲基溴化铵、鲸蜡硬脂基三甲基氯化铵、二硬脂基二甲基氯化铵、十八烷基二甲基苄基氯化铵、山嵛基三甲基溴化铵、苯扎氯铵、硬脂酸二乙基氨基乙酯、硬脂酸二甲基氨基丙酯、羊毛酯衍生物的季铵盐等。
两性表面活性剂可以包括羧基甜菜碱型、酰胺甜菜碱型、磺基甜菜碱型、羟基磺基甜菜碱型、酰胺磺基甜菜碱型、磷酰甜菜碱型、氨基羧酸盐型、咪唑啉衍生物型、酰胺胺型等。
有机或无机颜料可以包括无机颜料(例如,硅酸、硅酸酐、硅酸镁、滑石、绢云母、云母、高岭土、三氧化二铁、黏土、膨润土、钛涂覆云母、氯氧化铋、氧化锆、氧化镁、氧化锌、氧化钛、氧化铝、硫酸钙、硫酸钡、硫酸镁、碳酸钙、碳酸镁、铁氧化物、群青色、氧化铬、氢氧化铬、菱锌矿及其配合物);有机颜料(例如聚酰胺、聚酯、聚丙烯、聚苯乙烯、聚氨酯、乙烯基树脂、尿素树脂、酚醛树脂、氟树脂、硅树脂、丙烯酸类树脂、三聚氰胺树脂、环氧树脂、聚碳酸酯树脂、二乙烯基苯/苯乙烯共聚物、丝粉、纤维素、CI颜料黄和CI颜料橙);以及其无机颜料与有机颜料的复合颜料。
有机粉末可以包括金属皂例如硬脂酸钙;烷基磷酸金属盐(例如,棕榈酸锌钠、月桂酸锌皂和月桂酸钙);酰基氨基酸的多价金属盐(例如,N-月桂酰-β-丙氨酸钙、N-月桂酰-β-丙氨酸锌和N-月桂酰甘氨酸钙);酰胺磺酸的多价金属盐(例如,N-月桂酰-牛磺酸钙和N-棕榈酰基-牛磺酸钙);酰基碱性氨基酸(例如,N-ε-月桂酰-L-赖氨酸、N-ε-棕榈酰肼、N-α-帕里托酰鸟氨酸、N-α-月桂酰精氨酸和N-α-硬化牛脂脂肪酸酰基精氨酸);N-酰基多肽例如N-月桂酰甘氨酰甘氨酸;α-氨基脂肪酸例如α-氨基辛酸和α-氨基月桂酸;聚乙烯、聚丙烯、尼龙、聚甲基丙烯酸甲酯、聚苯乙烯、二乙烯基苯/苯乙烯共聚物、四氟乙烯等。
UV吸收剂可以包括对氨基苯甲酸、对氨基苯甲酸乙酯、对氨基苯甲酸戊酯、对氨基苯甲酸辛酯、水杨酸乙二醇酯、水杨酸苯酯、水杨酸辛酯、水杨酸苄酯、水杨酸丁基苯酯、水杨酸三甲环己酯、肉桂酸苄酯、对甲氧基肉桂酸2-乙氧基乙酯、对甲氧基肉桂酸辛酯、甘油单-2-乙基己烷二-对甲氧基肉桂酸酯、对甲氧基肉桂酸异丙酯、二异丙基/二异丙基肉桂酸酯混合物、尿刊酸乙酯、羟基甲氧基二苯甲酮、羟基甲氧基二苯甲酮磺酸及其盐、二羟基甲氧基二苯甲酮、二羟基甲氧基二苯甲酮二磺酸钠、二羟基二苯甲酮、四羟基二苯甲酮、4-叔丁基-4'-甲氧基二苯甲酰甲烷、2,4,6-三苯胺-p-(羰基-2'-乙基己基-1'-氧基)-1,3,5-三嗪、2-(2-羟基-5-甲基苯基)苯并三唑等。
杀菌剂可以包括扁柏酚、三氯生、三氯羟基二苯醚、葡糖酸洗必泰、苯氧乙醇、间苯二酚、异丙基甲基苯酚、薁、水杨酸、zincphyllione、苯扎氯铵、光敏剂第301号、单硝基愈创木酚钠、十一烯酸等。
抗氧化剂可以包括丁基羟基茴香醚、丙基没食子酸、异抗坏血酸等。
pH调节剂可以包括柠檬酸、柠檬酸钠、苹果酸、苹果酸钠、富马酸、富马酸钠、琥珀酸、琥珀酸钠、氢氧化钠、磷酸一氢钠等。
醇可以包括高级醇例如鲸蜡醇。
此外,可以添加的成分不限于以上组分,并且可以在不损害本发明的目的和效果的范围内使用以上成分中的任一者,条件是它们以基于组合物总重量的优选0.01重量%至5重量%,更优选0.01重量%至3重量%的量使用。
当本发明的制剂为洗剂、糊剂、乳膏剂或凝胶剂时,载体组分可以包括动物纤维、植物纤维、蜡、石蜡、淀粉、tracanth、纤维素衍生物、聚乙二醇、有机硅、膨润土、二氧化硅、滑石、氧化锌等。
当本发明的制剂为散剂或喷雾剂时,载体组分可以包括乳糖、滑石、二氧化硅、氢氧化铝、硅酸钙或聚酰胺粉末,并且特别地,当本发明的制剂为喷雾剂时,载体组分可以另外地包含推进剂(例如,氯氟烃、丙烷/丁烷或二甲醚)。
当本发明的制剂为溶液剂或乳剂时,载体组分可以包含溶剂、溶剂化剂或乳化剂,例如水、乙醇、异丙醇、碳酸乙酯、乙酸乙酯、苄醇、苯甲酸苄酯、丙二醇、1,3-丁基乙二醇油、甘油脂族酯、聚乙二醇或脂肪酸的脱水山梨糖醇酯。
当本发明的制剂为混悬剂时,载体组分可以包括液体稀释剂(例如,水、乙醇或丙二醇)、助悬剂(例如,乙氧基化异硬脂醇、聚氧乙烯山梨糖醇酯和聚氧乙烯脱水山梨糖醇酯)、微晶纤维素、偏氢氧化铝、膨润土、琼脂、tracanth等。
当本发明的制剂为含表面活性剂的清洁剂时,载体组分可以包括脂肪醇硫酸盐/酯、脂肪醇醚硫酸盐/酯、磺基琥珀酸单酯、羟乙磺酸盐/酯、咪唑啉莪衍生物、甲基牛磺酸盐、肌氨酸盐/酯、脂肪酸酰胺醚硫酸盐/酯、烷基酰胺甜菜碱、脂肪醇、脂肪酸甘油酯、脂肪酸二乙醇酰胺、植物油、亚麻精衍生物、乙氧基化甘油脂肪酸酯等。
在下文中,将通过工作实例更详细地描述本发明。以下实施例仅是帮助理解本发明的实例,并且本发明的范围不限于此。
发明实施方式
(实施方案)
实验例1.异原莪述烯醇的表皮生长因子受体(EGFR)的活性的测量
存在于作为细胞膜的EGF受体的EGFR的细胞质部分中的酪氨酸残基的磷酸化伴随EGFR的激活而发生。此外,通过利用当EGFR的活性持续得到保持时,EGFR被内化至细胞中的特征,在本实验使用可以测量EGFR活性的生物传感器系统的细胞(参见韩国特许专利公开第2019-0014340号)。
用10ng/ml的EGF和100μM的异原莪述烯醇(购自InterPharm)处理生物传感器系统的细胞,并观察荧光信号的变化。在将细胞在无血清培养基中培养24小时之后,用异原莪述烯醇处理细胞。
作为结果,如图1所示,确定与仅用溶剂DMSO处理的对照组相比,在用100μM的异原莪述烯醇处理的细胞中EGFR被激活。
实验例2.取决于异原莪述烯醇浓度的细胞毒性的测量
使用HaCaT细胞作为角质形成细胞测量异原莪述烯醇的细胞毒性。
当将细胞用为黄色水溶性底物的MTT四唑进行处理时,这样的底物被线粒体酶还原为MTT甲臜,并且颜色变为蓝紫色。也就是说,由于随着活细胞的数量的增加,蓝紫色显得更深,因此在540nm波长下的蓝紫色吸光度反映了活细胞的浓度。
具体地,在培养细胞至填满96孔板的每个孔之后,将细胞用浓度为0μM、10μM、25μM、50μM和100μM的异原莪述烯醇进行处理,并在37℃下在5% CO2培养箱中培养24小时和48小时。此后,使用MTT试剂测量细胞生存力。将未用异原莪述烯醇(0μM)处理的对照组用相同量的溶剂DMSO进行处理。
作为结果,如图2所示,确定当用100μM的异原莪述烯醇培养细胞48小时时,显示出一些细胞毒性,差异仅为10%。因此确定在角质形成细胞中的细胞毒性非常低。
实验例3.由于异原莪述烯醇的ERK和AKT活性
公知细胞外信号调节激酶(ERK)和蛋白激酶B(AKT)的活性通过EGFR的活性来传递细胞生长和存活所必需的信号。此外,在实验例1中,观察到在用于测量EGFR活性的生物传感系统的细胞中,由于EGFR的激活,异原莪述烯醇产生信号。因此,测量EGFR的活性是否由异原莪述烯醇诱导至ERK和AKT的活性。
具体地,将HaCaT细胞在无血清培养基中培养,分别用10μM的异原莪述烯醇处理10分钟、30分钟、和60分钟,此后,通过Western印迹分析磷酸化且活化的ERK和AKT。作为抗体,分别使用作为活化ERK和AKT的标志物的磷酸化抗体和用于测量其全部的通用抗体。
作为结果,如图3所示,确定10μM的异原莪述烯醇在10分钟内充分诱导ERK和AKT的活性。还确定ERK和AKT在30分钟内被迅速激活。
实验例4.由于异原莪述烯醇的基因表达变化
由EGFR产生的信号经由重要的信号介导蛋白质(例如ERK和AKT)诱导对细胞生长和存活重要的基因的表达。egr1、c-myc、c-jun和c-fos基因受EGFR调控,并且公知作为促进细胞生长和存活的基因。在实验例2中,将在无血清培养基中培养的HaCaT细胞中用浓度为10μM的异原莪述烯醇(其充分诱导ERK和AKT活性)进行处理,并在1小时之后,分析各基因表达变化。
具体地,使用Tri-试剂用于mRNA提取,使用poly-A引物和逆转录酶通过逆转录聚合酶链反应(RT-PCR)产生cDNA,并且使用实时定量PCR来测量各mRNA的量。各mRNA的量用β-肌动蛋白(其为组成基因)的量进行校正。
作为结果,如图4所示,egr1、c-myc、c-jun和c-fos mRNA增加。
实验例5.通过异原莪述烯醇的角质形成细胞的生长和增殖的测量
细胞计数试剂盒-8(Cell counting kit-8,CCK-8)是称为WST-8的水溶性四唑盐试剂,并且通过活细胞的线粒体中的脱氢酶形成橙色水溶性甲臜。由此,在450nm(其为由CCK-8显示的橙色波长)下的吸光度反映了细胞的数量,并因此用于测量细胞的生长和增殖。通过CCK-8测定,通过使用浓度为10μM或更小(即1nM、10nM、100nM、1μM和10μM)的无细胞毒性的异原莪述烯醇测量作为角质形成细胞的HaCaT细胞的生长促进作用。
具体地,在将1×104个HaCaT细胞在96孔板中培养24小时之后,将细胞换成无血清培养基以除去培养基中包含的生长因子,并用浓度为1nM、10nM、100nM、1μM和10μM的异原莪述烯醇进行处理。作为阴性对照组,使用相同量的溶剂DMSO,以及作为阳性对照组,使用浓度为1ng/ml的EGF。在培养24小时之后,用CCK8处理细胞,并通过测量在450nm下的吸光度分析细胞生长。
作为结果,如图5所示,确定当用10nM、100nM、1μM和10μM浓度的异原莪述烯醇处理细胞时,与阴性对照组相比,细胞的生长和增殖显著增加(单因素ANOVA,*p<0.05)。
实验例6.异原莪述烯醇的细胞生长促进作用(伤口愈合评价)
异原莪述烯醇的细胞生长促进作用通过伤口愈合评价来确定。具体地,在将HaCaT细胞在12孔板中充分培养之后,使用黄色尖端刮掉一定区域上的细胞以形成伤口区域,换成无血清培养基,并用1μM的异原莪述烯醇处理。作为阴性对照组,使用溶剂DMSO,以及作为阳性对照组,使用1ng/ml的EGF。
作为结果,如图6a和6b所示,确定与阴性对照组相比,当在培养6天期间异原莪述烯醇处理时,伤口区域被更快地填充,并且从第3天起,与阴性对照组相比显示出更显著的差异。(t-测试,*p<0.05)。
实验例7.异原莪述烯醇的关于抗紫外线(UVB)刺激的效力
在紫外线波长中,使对应于280nm至315nm的UVB波长穿透其中存在许多角质形成细胞的表皮层以引起细胞损伤。由于由UVB引起的细胞损伤也导致凋亡,因此确定异原莪述烯醇是否表现出对由UVB引起的细胞损伤的抗性(由于其促进角质形成细胞的生长和存活信号的传递)。
具体地,将其中培养有1×104个HaCaT细胞的96孔板用500mJ/cm2的UVB照射,然后在包含浓度为1μM的异原莪述烯醇或EGF的无血清培养基中培养。使用溶剂DMSO作为阴性对照组,并使用实验例6中使用的CCK-8法测量细胞生存力。
作为结果,如图7所示,确定在阴性对照组中由于500mJ/cm2的UVB引起凋亡,使得仅约40%的细胞存活,而由于1μM的异原莪述烯醇,约60%的细胞存活,这显示出统计学上显著的值(t-测试,*p<0.05)。
工业适用性
虽然以上已经示出和描述了示例性实施方案,但对于本领域技术人员将明显的是,可以在不脱离如由所附权利要求限定的本公开内容的范围的情况下进行修改和变化。
Claims (9)
2.根据权利要求1所述的用于防老化或皮肤再生的组合物,以10nM至100μM的量包含异原莪述烯醇或其盐。
3.根据权利要求1所述的用于防老化或皮肤再生的组合物,用于改善或治疗由烧伤引起的皮肤和组织损伤。
4.根据权利要求1所述的用于防老化或皮肤再生的组合物,用于防止或改善来自外部刺激的细胞损伤。
5.根据权利要求4所述的用于防老化或皮肤再生的组合物,其中所述外部刺激为紫外线。
6.根据权利要求1所述的用于防老化或皮肤再生的组合物,用于增强皮肤弹性。
7.根据权利要求1所述的用于防老化或皮肤再生的组合物,用于改善皮肤皱纹的问题。
8.根据权利要求1至7中任一项所述的用于防老化或皮肤再生的组合物,其中所述组合物为药物组合物、健康功能性食品组合物或化妆品组合物。
9.由下式(1)表示的异原莪述烯醇或其盐用于制备用于防老化或皮肤再生的组合物的用途。
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