CN117940103A - 抗细纹和皱纹的局部用组合物 - Google Patents
抗细纹和皱纹的局部用组合物 Download PDFInfo
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- CN117940103A CN117940103A CN202280058123.3A CN202280058123A CN117940103A CN 117940103 A CN117940103 A CN 117940103A CN 202280058123 A CN202280058123 A CN 202280058123A CN 117940103 A CN117940103 A CN 117940103A
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- oil
- wrinkles
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Abstract
公开了局部皮肤用组合物及其使用方法。该组合物可以包含至少一种能够处理皮肤状况的皮肤活性成分,和多种能够增加皮肤活性成分渗透到皮肤中的皮肤渗透促进剂。多种皮肤渗透促进剂可以包括乙醇、异山梨醇二甲基醚、戊二醇、乙氧基二甘醇和丙二醇。
Description
发明背景
相关申请的交叉引用
本申请要求2021年7月1日提交的美国临时专利申请序列第63/217,625号的优先权权益,其通过引用整体并入本文。
A.发明领域
本发明一般涉及能够减轻人皮肤上细纹或皱纹的外观的化妆品皮肤护理组合物。该组合物可以包含至少一种皮肤活性成分和多种能够增加皮肤活性成分渗透到皮肤中的皮肤渗透促进剂。
B.相关技术说明
由于重复性肌肉收缩,面部皮肤随着时间出现某些类型的皱纹。通常把这些类型的皱纹称为面部皱纹,其实例可以包括皱眉纹或眉间纹、前额纹、鱼尾纹、兔子纹或鼻纹、有窝的下巴、微笑提拉纹、唇纹等。目前,可以使用多种可商购获得的处理物质,并据称作为面部肌肉松弛剂/肌肉收缩抑制剂是有效的。据说这些物质能够减轻这种皱纹的外观。这种处理物质的非限制性实例包括可注射的神经调质例如(onabotulinumtoxinA)、(abobotulinumtoxinA)或/>(incobotulinumtoxinA)。其他实例包括局部施用的处理物质如化学化合物(例如γ-氨基丁酸)和肽(例如乙酰基六肽-3)。
与可注射的神经调质有关的问题,例如过敏反应、皮疹、瘙痒、注射部位反应(淤斑、出血、疼痛、红肿、肿胀或压痛)、肌肉僵硬、发烧、咳嗽、咽喉痛、流感样症状、颈部或背部疼痛和从注射部位扩散到身体其它部位的可能,会造成严重的风险包括说话困难、吞咽或呼吸困难、肌无力、睑下垂和视力模糊或复视。类似地,化学化合物具有其自身缺点,例如皮肤刺激和潜在过敏反应。除了与可注射的神经调质和化学化合物相关的生理问题之外,对于大部分受益人群来说,这些选择性美容手术的成本高昂。
已经有尝试来生产能够放松面部肌肉的天然成分。虽然这些天然成分可以提供一些肌肉松弛效果,但经常会有将这些成分输送到皮肤中的问题。特别是,有效量的天然成分可能不能渗透到皮肤表面和/或输送到皮肤的表皮层或真皮层或这些皮肤层内或下面的肌肉细胞。
因此,先前改善皮肤视觉外观的尝试已经显示出具有各种缺点,例如高成本、医疗风险、皮肤刺激、恢复期延长或未达到承诺的皮肤益处。
发明内容
已经确定了至少一个上述问题的解决方案。在本发明的一个方面,该解决方案可以包括使用皮肤渗透促进剂的组合,其可以增加皮肤活性成分向皮肤的表皮和/或真皮层和/或存在于这些层内的肌肉细胞或位于这些皮肤层下面的肌肉细胞的递送。能够增加皮肤活性成分渗透到皮肤中的皮肤渗透促进剂的组合可以包括乙醇、异山梨醇二甲基醚、戊二醇、乙氧基二甘醇和丙二醇。在一些具体实施方案中,本发明的组合物可以包含5重量%至15重量%的乙醇、5重量%至15重量%的异山梨醇二甲基醚、3重量%至7重量%的戊二醇、1重量%至5重量%的乙氧基二甘醇和1重量%至5重量%的丙二醇。
在本发明的一个方面,公开了改善人的皮肤外观的方法。在一些实施方案中,该方法包括局部施用局部皮肤用组合物,该组合物包含至少一种能够处理皮肤状况的皮肤活性成分和多种能够增加皮肤活性成分渗透到皮肤中的皮肤渗透促进剂。在一些实施方案中,多种皮肤渗透促进剂包括乙醇、异山梨醇二甲基醚、戊二醇、乙氧基二甘醇和丙二醇。在一些实施方案中,至少一种皮肤活性成分能够减少人面部肌肉的肌肉收缩和/或减轻人皮肤上细纹或皱纹的外观。在一些实施方案中,至少一种皮肤活性成分是千日菊(Acmellaoleracea)提取物、法国薰衣草(Lavendula stoechas)提取物或迷迭香(Rosmarinusofficinalis)叶提取物。在一些实施方案中,组合物包含千日菊提取物、法国薰衣草提取物和迷迭香叶提取物。在一些实施方案中,组合物包含0.02重量%至2重量%的千日菊提取物、0.01重量%至1重量%的法国薰衣草提取物和0.005重量%至0.5重量%的迷迭香叶提取物。在一些实施方案中,千日菊提取物刺激了皮肤中胶原蛋白的产生和层黏连蛋白的产生。在一些实施方案中,迷迭香叶提取物减少了皮肤中基质金属蛋白酶1、基质金属蛋白酶3或基质金属蛋白酶9的产生。
在一些实施方案中,每种提取物分别为水提取物、乙醇提取物、多元醇提取物或其组合。在一些实施方案中,迷迭香叶提取物用包含甜菜碱、乳酸和水的流体提取溶剂混合物获得。在一些实施方案中,法国薰衣草提取物是用超临界二氧化碳(CO2)提取溶剂获得的法国薰衣草花/叶/茎提取物。在一些实施方案中,千日菊提取物用包含水、乙醇和1,3丙二醇的流体提取溶剂混合物获得。
在一些实施方案中,组合物包含5重量%至15重量%的乙醇、5重量%至15重量%的异山梨醇二甲基醚、3重量%至7重量%的戊二醇、1重量%至5重量%的乙氧基二甘醇和1重量%至5重量%的丙二醇。在一些实施方案中,组合物包含30重量%到60重量%的水。在一些实施方案中,该组合物还包含鲸蜡酯、硬脂醇、鲸蜡硬脂醇橄榄油酸酯、失水山梨醇橄榄油酸酯、丙烯酸酯类/C10-30烷醇丙烯酸酯交联聚合物、鲸蜡醇棕榈酸酯和失水山梨醇棕榈酸酯。在一些实施方案中,组合物还包含3重量%至7重量%的鲸蜡酯、1重量%至5重量%的硬脂醇、1重量%至5重量%的鲸蜡硬脂醇橄榄油酸酯、1重量%至5重量%的失水山梨醇橄榄油酸酯、0.1重量%至1重量%的丙烯酸酯类/C10-30烷醇丙烯酸酯交联聚合物、0.1重量%至1重量%的鲸蜡醇棕榈酸酯和0.1重量%至1重量%的失水山梨醇棕榈酸酯。
在一些实施方式中,组合物为乳液。在一些实施方案中,乳液是水包油乳液。在一些实施方案中,将组合物施用至细纹或皱纹上。在一些实施方案中,在局部施用后减轻了细纹或皱纹的外观。在一些实施方案中,该组合物减少了引起细纹或皱纹外观的肌肉收缩。在一些实施方案中,面部肌肉是眉间复合肌、眼轮匝肌、降肌或额肌或其任意组合。在一些实施方案中,细纹或皱纹是面部皱纹。在一些实施方案中,面部皮肤是前额皮肤、脸颊皮肤、下巴皮肤和/或眼眶区域皮肤。
在一些实施方案中,将组合物直接施用至对象的皮肤。在一些实施方案中,将该组合物施用至对象的皮肤上将增加皮肤紧致度和张力,减少氧化应激,减少皮肤自由基含量,减轻细纹和/或皱纹的外观,减少皮肤松垂,减少慢性老化和光老化的皮肤病学迹象,最小化疲劳和/或压力的皮肤病学迹象,和/或使对象的皮肤再生。在一些实施方案中,组合物以乳液的形式提供。
本公开的一些方面涉及改善皮肤状况或外观,其包括将本文公开的任何一种组合物施用至有需要的皮肤。在一个方面,将本文公开的任何一种组合物施用于皮肤,并将该组合物留在皮肤上,或在一段时间后从皮肤上除去。在一些方面,本文公开的组合物用于处理和/或减少皱纹。在一些方面,本文公开的组合物用于增加皮肤紧致度和张力。在一些方面,本文公开的组合物用于降低氧化应激和皮肤自由基含量。
在一些方面,将本发明的组合物配制为局部皮肤用组合物。组合物可以配制为局部皮肤用组合物,其在使用期间每天施用至少1次、2次、3次、4次、5次、6次、7次或多于7次。在本发明的其他方面,组合物可以是储存稳定的。还预期可以例如根据理想组合物的类型,选择组合物的溶解程度以实现理想的结果。在一些实施方案中,将组合物直接施用至皮肤。
在一些方面,本发明的组合物还可以包含表面活性剂、含硅酮的化合物、UV剂、油和/或本说明书所确定的其他成分或本领域中已知的成分。该组合物可以是爽肤水、乳膏、身体乳、面膜、擦洗剂、清洁剂、霜剂、爽肤水、洗剂、凝胶、精华素、乳液(例如,水包油、油包水、水包硅酮、硅酮包水、水包油包水、油包水包油、硅酮包水包油等)、溶液(如水溶液或水醇溶液)、无水基质(如,口红或粉剂)、软膏剂、乳剂、糊状物、气溶胶、固体形式、眼部凝胶、凝胶精华、凝胶乳液等。该组合物可以配制成用于局部皮肤施用,在使用期间每天施用至少1次、2次、3次、4次、5次、6次、7次或多于7次。在本发明的一些方面,组合物可以是储存稳定或颜色稳定的,或是两者。还预期可以选择组合物的黏度以达到希望的结果,例如根据希望的组合物类型,这种组合物的黏度可以为约1cp至远超过1百万cp,或者是其间可获得的任意范围或整数(例如,在25℃下在布氏黏度计上用TC转子以2.5rpm测得的2cp、3cp、4cp、5cp、6cp、7cp、8cp、9cp、10cp、20cp、30cp、40cp、50cp、60cp、70cp、80cp、90cp、100cp、200cp、300cp、400cp、500cp、600cp、700cp、800cp、900cp、1000cp、2000cp、3000cp、4000cp、5000cp、6000cp、7000cp、8000cp、9000cp、10000cp、20000cp、30000cp、40000cp、50000cp、60000cp、70000cp、80000cp、90000cp、100000cp、200000cp、300000cp、400000cp、500000cp、600000cp、700000cp、800000cp、900000cp、1000000cp、2000000cp、3000000cp、4000000cp、5000000cp、10000000cp等)。
在非限制性方面,组合物的pH可以为约6至约9。在一些方面,pH可以为1、2、3、4、5、6、7、8、9、10、11、12、13或14。组合物可以包含甘油三酯。非限制性实例包括短链、中链和长链甘油三酯。在某些方面,甘油三酯是中链甘油三酯(例如辛酸癸酸甘油三酯)。组合物还可以包含防腐剂。防腐剂的非限制性实例包括苯氧乙醇、对羟基苯甲酸甲酯、对羟基苯甲酸丙酯、碘丙炔醇丁基氨甲酸酯、山梨酸钾、苯甲酸钠或其任意混合物。在一些实施方案中,组合物不含对羟基苯甲酸酯。
本发明的组合物可以具有UVA和UVB吸收性质。该组合物可以具有2、3、4、5、6、7、8、9、10、11、12、13、14、15、20、25、30、35、40、45、50、55、60或大于60,或是其间任意整数或可获得的防晒指数(SPF)。该组合物可以是防晒乳液、喷雾剂、霜剂、爽肤水、精华、清洁剂或面膜。
本发明的组合物还可包含以下附加成分中的任何一种、任意组合或全部:调理剂、保湿剂、pH调节剂、结构化剂、无机盐、防腐剂、增稠剂、含硅酮的化合物、精油、香精、维生素、药物成分或抗氧化剂,或上述成分的任意组合或上述成分的混合物。在某些方面,组合物可以包含前述确定的这些附加成分中的至少二种、三种、四种、五种、六种、七种、八种、九种、十种或多于十种、或全部。这些附加成分的非限制性实例在本说明书全文确定并通过引用并入本部分。这类成分的量的范围以组合物的重量或体积计可以为0.0001%至99.9%,或者是在如本说明书其它部分中所公开的范围之间的任何整数或范围,其通过引用并入本段。
还预期本说明书全文所公开的组合物可以用作免洗型或洗去型组合物。举例来说,免洗型组合物可以是局部施用至皮肤并在皮肤上保留一段时间(例如,至少5分钟、6分钟、7分钟、8分钟、9分钟、10分钟、20分钟或30分钟,或至少1小时、2小时、3小时、4小时、5小时、6小时、7小时、8小时、9小时、10小时、11小时、12小时、13小时、14小时、15小时、16小时、17小时、18小时、19小时、20小时、21小时、22小时、23小时或24小时,或者整夜或全天)的一种组合物。或者,洗去型组合物可以是旨在施用于皮肤然后在一段时间内,例如小于5分钟、4分钟、3分钟、2分钟或1分钟,(例如用水)从皮肤除去或洗去的产品。洗去型组合物的实例可以是皮肤清洁剂、洗发水、护发素或肥皂。免洗型组合物的实例可以是皮肤保湿剂、防晒剂、面膜、晚霜或日霜。
还预期了包含本发明的组合物的试剂盒。在特定的实施方案中,组合物包含在容器中。容器可以是瓶子、分配器或包装。该容器可以分配预定量的组合物。在某些方面,组合物以喷雾、薄雾、团块或液体形式分配。容器可以在其表面上包含标记。该标记可以是单词、缩写、图片或符号。
预期对于本发明的任何方法或组合物,可以实施本说明书中所讨论的任意实施方案,反之亦然。此外,本发明的组合物可以用于实现本发明的方法。
在一个实施方案中,本发明的组合物可以是药用舒适的或化妆用舒适的,或可以具有舒适的触觉性质。“药用舒适的”、“化妆用舒适的”和/或“舒适的触觉性质”描述了具有令皮肤感觉舒适的特定的触觉性质的组合物(例如,不太水或太油的组合物、具有丝滑质地的组合物、非黏性或非黏着的组合物等)。药用舒适的或化妆用舒适的还可以涉及组合物的乳脂状或润滑性质,或组合物的保湿性质。
还公开了本发明的以下方面1至21。
方面1是局部皮肤用组合物,其包含至少一种能够处理皮肤状况的皮肤活性成分和多种能够增加皮肤活性成分渗透到皮肤中的皮肤渗透促进剂,其中多种皮肤渗透促进剂包括:乙醇;异山梨醇二甲基醚;戊二醇;乙氧基二甘醇;和丙二醇。方面2是方面1的局部皮肤用组合物,其中至少一种皮肤活性成分能够减少人面部肌肉的肌肉收缩和/或减轻人皮肤上细纹或皱纹的外观。方面3是方面1或2的局部皮肤用组合物,其中至少一种皮肤活性成分是千日菊提取物、法国薰衣草提取物或迷迭香叶提取物。方面4是方面1至3中任一方面的局部皮肤用组合物,其中组合物包含千日菊提取物、法国薰衣草提取物和迷迭香叶提取物。方面5是方面4的局部皮肤用组合物,其中组合物包含0.02重量%至2重量%的千日菊提取物、0.01重量%至1重量%的法国薰衣草提取物和0.005重量%至0.5重量%的迷迭香叶提取物。方面6为方面3至5中任一方面的局部皮肤用组合物,其中每种提取物分别为水提取物、乙醇提取物、多元醇提取物或其组合。方面7是方面3至5中任一方面的局部皮肤用组合物,其中:迷迭香叶提取物用包含甜菜碱、乳酸和水的流体提取溶剂混合物获得;法国薰衣草提取物是用超临界二氧化碳(CO2)提取溶剂获得的法国薰衣草花/叶/茎提取物;千日菊提取物用包含水、乙醇和1,3丙二醇的流体提取溶剂混合物获得。方面8是方面1至7中任一方面的局部皮肤用组合物,其中该组合物包含:5重量%至15重量%的乙醇、5重量%至15重量%的异山梨醇二甲基醚、3重量%至7重量%的戊二醇、1重量%至5重量%的乙氧基二甘醇和1重量%至5重量%的丙二醇。方面9是方面1至8中任一方面的局部皮肤用组合物,其中组合物包含30重量%到60重量%的水。方面10是方面1至9中任一方面的局部皮肤用组合物,其中该组合物还包含:鲸蜡酯、硬脂醇、鲸蜡硬脂醇橄榄油酸酯、失水山梨醇橄榄油酸酯、丙烯酸酯类/C10-30烷醇丙烯酸酯交联聚合物、鲸蜡醇棕榈酸酯和失水山梨醇棕榈酸酯。方面11是方面10的局部皮肤用组合物,其中该组合物还包含:3重量%至7重量%的鲸蜡酯、1重量%至5重量%的硬脂醇、1重量%至5重量%的鲸蜡硬脂醇橄榄油酸酯、1重量%至5重量%的失水山梨醇橄榄油酸酯、0.1重量%至1重量%的丙烯酸酯类/C10-30烷醇丙烯酸酯交联聚合物、0.1重量%至1重量%的鲸蜡醇棕榈酸酯和0.1重量%至1重量%的失水山梨醇棕榈酸酯。方面12是方面1至11中任一方面的局部皮肤用组合物,其中该组合物是乳液。方面13是方面12的局部皮肤用组合物,其中乳液是水包油乳液。方面14是改善人的皮肤外观的方法,该方法包括将方面1至13中任一方面的组合物局部施用至人的皮肤上,其中该组合物改善了皮肤的外观。方面15是方面14的方法,其中将组合物施用至细纹或皱纹上。方面16是方面15的方法,其中在局部施用后减轻了细纹或皱纹的外观。方面17是方面16的方法,其中组合物减少引起细纹或皱纹外观的肌肉收缩。方面18是方面17的方法,其中面部肌肉是眉间复合肌、眼轮匝肌、降肌、或额肌或其任意组合。方面19是方面15至18中任一方面的方法,其中细纹或皱纹是面部皱纹。方面20是方面14至19中任一方面的方法,其中该组合物包括:千日菊提取物,其中千日菊提取物刺激皮肤中胶原蛋白的产生和层黏连蛋白的产生;和迷迭香叶提取物,其中迷迭香叶提取物减少皮肤中基质金属蛋白酶1、基质金属蛋白酶3或基质金属蛋白酶9的产生。方面21是方面14至20中任一方面的方法,其中面部皮肤是前额皮肤、脸颊皮肤、下巴皮肤或眼眶区域皮肤。
“局部施用”是指将组合物施用至皮肤表面上。这类组合物一般为皮肤病学上可接受的,这是因为当施用至皮肤时,其不具有异常毒性、不相容性、不稳定性、过敏反应等。可以将本发明的局部皮肤用组合物配制为达到目标溶解度,以避免在施用至皮肤后明显滴落。短语“减轻细纹和/或皱纹的外观”是指降低面部细纹和皱纹,脸颊、前额上的面部皱纹,眼睛之间的垂直皱纹,眼睛上方的横向皱纹,和嘴巴周围的木偶纹,特别是很深的皱纹或皮褶的可察觉性;减轻和/或减少细纹和/或皱纹的外观和/或深度;和/或改善眼眶下的细纹和/或眶周细纹的外观。
术语“约”或“近似”定义为本领域普通技术人员所理解的接近于。在一个非限制性实施方案中,该术语定义为偏差在10%以内,优选偏差在5%以内,更优选偏差在1%以内,最优选偏差在0.5%以内。
术语“基本上”及其变体是指为包含偏差在10%以内、偏差在5%以内、偏差在1%以内、或偏差在0.5%以内的范围。
术语“抑制”或“减少”或这些术语的任何变体包括为了达到预期结果的任何可测量的减少或完全的抑制。术语“促进”或“增加”或这些术语的任何变体包括为了达到预期结果的任何可测量的蛋白质或分子(例如,基质蛋白如胶原蛋白或弹性蛋白)的增加或产生。
如本说明书和/或权利要求所使用的,术语“有效的”表示足以实现期望的、预期的或理想的结果。
当在权利要求和/或说明书中与术语“包含”、“包括”、“具有”或“含有”或这些术语的任何变体的一起使用时,要素前面不使用数量词可以表示“一个”,但是其也符合“一个或更多个”、“至少一个”和“一个或多于一个”的含义。
如本说明书和权利要求所使用的,单词“包含”、“具有”、“包括”或“含有”是包括性的或开放式的,并且不排除附加的、未列举的要素或方法步骤。
使用的组合物和方法可以“包含”、“基本上组成为”或“组成为”本说明书通篇所公开的任意成分或步骤。关于短语“基本上组成为”,本发明的组合物和方法的基本和新颖性质是具有皮肤渗透促进剂的组合的组合物,以增加皮肤活性成分向皮肤中的递送(例如,向皮肤的表皮或真皮层和/或向存在于这些层中或之下的肌肉细胞中的递送)。
如本文所述,术语“皱纹”是皮肤上的褶皱、脊、皮褶、沟、痘痕、凹坑或凹陷区域,其可以是由习惯性的面部表情、由于衰老导致的胶原蛋白和/或弹性蛋白的流失、晒伤、吸烟、水分不足以及各种其他因素引起的。皱纹可以是很深的皮褶至细纹。本文考虑了出现在身体的任何部分上的皱纹,特别是对象的头部或颈部上的皱纹。可以根据本公开处理的皱纹包括但不限于额纹、鱼尾纹、鼻唇沟、眼下或眉间的一种或多于一种细纹及其组合。由于重复的面部肌肉收缩引起的动态面部皱纹和/或前额皱纹可以包括面部皱纹。
本文所用的“处理”是指暂时或永久减轻(或消除)皮肤瑕疵的一个或多于一个特征。当施用该组合物来处理伤口时,与没有施用的伤口相比,该组合物促进了正常愈合。也就是说,经处理的伤口的尺寸(长度、深度、高度和/或宽度)、特征、颜色和/或质地更接近正常的、未受伤的组织。在该点上,用所公开的组合物处理伤口可以防止、最小化或改善由伤口愈合引起的疤痕形成的外观。此外,当施用所公开的组合物来处理皱纹时,如果皱纹的外观或突出程度在视觉上或临床上减少,则皱纹得到处理。也就是说,与处理前的皱纹相比,长度和/或深度减小了。或者,处理可以包括防止皱纹。在该点上,所公开的组合物可以被施用至通常产生皱纹的皮肤区域,例如前额、嘴唇、眼睑、鼻唇沟、眼睛下方的皮肤或眉间,以防止皱纹的产生。
通过下面的详细描述,本发明的其他目的、特征和优点会变得明显。然而,应理解详细的描述和实施例在表明本发明的具体实施方案时仅以举例说明给出。另外,期望通过该详细描述,本发明的精神和范围内的变化和修改对于本领域技术人员会变得明显。
附图简要说明
以下附图形成本说明书的一部分,并被包含以进一步证实本发明的特定方面。通过参照一个或多于一个这些附图结合本文所示的说明书实施方案的详细说明可以更好地理解本发明。
图1提供了表明一组人的眉间区域肌肉收缩的EMG响应面积(mV.s)相对于基线(第0周)改善的平均%的数据,所述一组人使用了具有迷迭香叶提取物、法国薰衣草提取物和千日菊提取物的组合的本发明的组合物。改善的平均%表明了眉间区域肌肉收缩的减少随时间(第2周、4周和8周)的变化。“*”表示p<0.05,当与基线(第0周)相比,改善具有统计学显著性。
图2提供了表明一组人的眉间区域肌肉收缩的EMG响应面积(mV.s)相对于基线(第0周)显示出改善的人的平均%的数据,所述一组人使用了具有迷迭香叶提取物、法国薰衣草提取物和千日菊提取物的组合的本发明的组合物。显示出改善的人的平均%表明眉间区域肌肉收缩的减少随时间(第2周、4周和8周)的变化。
图3提供了表明一组人细纹或皱纹的外观减轻相对于基线(第0周)改善的平均%的数据,所述一组人使用了具有迷迭香叶提取物、法国薰衣草提取物和千日菊提取物的组合的本发明的组合物。改善的平均%表明了经处理的区域中细纹或皱纹的外观减轻随时间(第2周、4周和8周)的变化。“*”表示p<0.05,当与基线(第0周)相比,改善具有统计学显著性。
图4提供了表明一组人细纹或皱纹的减少相对于基线(第0周)显示出改善的人的平均%的数据,所述一组人使用了具有迷迭香叶提取物、法国薰衣草提取物和千日菊提取物的组合的本发明的组合物。显示出改善的人的平均%表明了经处理的区域中细纹或皱纹的外观减少随时间(第2周、4周和8周)的变化。
图5提供了数据,表明一组人眉间区域肌肉收缩的EMG响应面积(mV.s)相对于基线(第0周)改善的平均%,所述一组人使用了具有迷迭香叶提取物、法国薰衣草提取物和千日菊提取物的组合的本发明的组合物,并且对比了使用微型滚轮施用所述组合物与不使用微型滚轮施用所述组合物。改善的平均%表明当使用微型滚轮来施用组合物时,第2周、4周和8周时面部肌肉收缩的减少与基线相比较快且较强。“*”表示p<0.05,当与基线(第0周)相比,改善具有统计学显著性。
图6提供了表明一组人的眉间区域肌肉收缩的EMG响应面积(mV.s)相对于基线(第0周)显示出改善的人的平均%的数据,所述一组人使用了具有迷迭香叶提取物、法国薰衣草提取物和千日菊提取物的组合的本发明的组合物。显示出改善的人的平均%表明当使用微型滚轮来施用组合物时,眉间区域在第2周、4周和8周时面部肌肉收缩的减少较快且较强。
图7提供了表明一组人细纹或皱纹的外观减少相对于基线(第0周)改善的平均%的数据,所述一组人使用了具有迷迭香叶提取物、法国薰衣草提取物和千日菊提取物的组合的本发明的组合物。改善的平均%表明当使用微型滚轮施用组合物和不使用微型滚轮施用组合物时,经处理的区域中细纹和皱纹的外观减少随时间(第2周、4周和8周)的变化。“*”表示p<0.05,当与基线(第0周)相比,改善具有统计学显著性。
图8提供了表明一组人细纹或皱纹的减少相对于基线(第0周)改善的人的平均%的数据,所述一组人使用了具有迷迭香叶提取物、法国薰衣草提取物和千日菊提取物的组合的本发明的组合物。显示出改善的人的平均%表明当使用微型滚轮施用组合物和不使用微型滚轮施用组合物时,经处理的区域中细纹和皱纹的外观减少随时间(第2周、4周和8周)的变化。
图9提供了表明一组人的皮肤纹理/粗糙度相对于基线(第0周)改善的平均%的数据,所述一组人使用了具有迷迭香叶提取物、法国薰衣草提取物和千日菊提取物的组合的本发明的组合物。改善的平均%表明当使用微型滚轮施用组合物和不使用微型滚轮施用组合物时,经处理的区域中经改善的皮肤纹理/粗糙度随时间(第2周、4周和8周)的变化。“*”表示p<0.05,当与基线(第0周)相比,改善具有统计学显著性。
图10提供了表明一组人的皮肤纹理/粗糙度相对于基线(第0周)显示出改善的人的平均%的数据,所述一组人使用了具有迷迭香叶提取物、法国薰衣草提取物和千日菊提取物的组合的本发明的组合物。显示出改善的人的平均%表明当使用微型滚轮施用组合物和不使用微型滚轮施用组合物时,经处理的区域中经改善的皮肤纹理/粗糙度随时间(第2周、4周和8周)的变化。
详细描述
如上所述,本发明提供了与当前化妆品相关的问题的解决方案。该解决方案的前提是使用多种成分的组合来减少或逆转皱纹和其他与衰老相关的迹象。皮肤起皱和松弛或松动是影响皮肤外观的两个主要事件。在眶周、眉间、前额和口周区域的面部表情肌肉涉及各种表情,包括微笑、皱眉、眯眼和撅嘴。这些肌肉的活动对其覆盖的皮肤施加了比在面部其他区域更大的物理应力。当这些肌肉在皮肤下方收缩,然后放松并回到其原来的放松长度时,形成了皱纹。覆盖的皮肤还可以缩短和回弹,但是肌肉不可以。因此,当肌肉收缩时,皮肤往往会弯屈、向内折并形成皱纹。本文公开的组合物包含松弛皱纹的组分,以对抗与衰老相关的皮肤疲软和起皱。组合物还包含抗氧化剂,以保护皮肤免于由紫外光和其他环境因素引起的氧化应激。
以下子章节更详细地描述本发明的非限制性方面。
A.渗透促进剂
本文公开的组合物包含多种能够增加皮肤活性成分渗透到皮肤中的皮肤渗透促进剂。皮肤渗透促进剂包括乙醇、异山梨醇二甲基醚、戊二醇、乙氧基二甘醇和丙二醇。在化妆品中,乙醇因其高挥发性(涂抹后立即消失)、干燥、清爽和抗菌性质而被使用。乙醇有助于活性成分渗透到皮肤表层,并促进成分在皮肤上的沉积。异山梨醇二甲基醚是溶剂和载体,其为个人护理产品中的活性成分提供了安全、有效的递送增强机制。当用作渗透促进剂时,异山梨醇二甲基醚允许制剂师减少达到效果所需的活性物质的量,从而降低制剂成本,并降低攻击性活性成分对皮肤的刺激。戊二醇是保湿剂,其具有保湿和抗菌性质。戊二醇是可溶于水的5碳有机化合物,可用于增强皮肤对有机化合物的吸收。乙氧基二甘醇促进活性有机成分溶解至制剂中,有助于关键成分更有效地发挥作用。乙氧基二甘醇还能改善制剂的质地或厚度,使它们感觉更轻盈,更容易涂抹。丙二醇是保湿剂,添加到化妆品中以增加皮肤的保湿性。当组合使用时,这些成分充当活性成分的载体。它们容易被皮肤吸收并增强皮肤中所含活性成分的吸收。
B.活性成分
千日菊提取物是来自千日菊植物的提取物。千日菊可以在南美、马达加斯加和马斯克林群岛发现。在优选的情况下,千日菊提取物来自千日菊的花、叶和茎部分的组合。这些部分可以混合然后捣碎或浸渍,或者将其捣碎或浸渍然后进行组合。然后,可以对得到的经捣碎或经浸渍的花/叶/茎材料进行水-醇(优选水-乙醇)提取过程或水-醇-多元醇提取过程。在优选的情况中多元醇可以为1,3-丙二醇。醇可以从所得提取物中除去。然后,千日菊提取物可用于本发明的组合物中。在优选的情况下,千日菊花/叶/茎提取物可以从水-乙醇-1,3-丙二醇溶剂混合物中获得。然后,得到的提取物可用于本发明的组合物中或还可以进行加工以除去乙醇,然后可以用于本发明的组合物中。或者,提取溶剂可以是水和多元醇的组合,优选1,3-丙二醇,不含乙醇。反应混合物中存在的水、乙醇和/或多元醇的量可以按所需来改变。在一些情况下,千日菊可商购获得。在一些情况下,千日菊提取物可以以商品名EXPRESSION AF由Gattefossé(法国)所提供。在本发明的上下文中发现,千日菊提取物可以通过减少或抑制肌管中的钙流入和减少或防止动作电位产生,来减少或抑制肌管收缩。还发现了千日菊提取物可以增加皮肤细胞(例如角质细胞或人真皮成纤维细胞)中胶原蛋白的产生和层黏连蛋白的产生。千日菊提取物包括镇痛烷基酰胺,如能放松面部肌肉和减轻肌肉紧张的千日菊酰胺。
法国薰衣草提取物是来自植物法国薰衣草的提取物。法国薰衣草原产自地中海地区,是常绿灌木,高约30cm至100cm,叶为浅灰色且被绒毛,叶长约1cm至4cm。在优选的情况下,法国薰衣草提取物来自法国薰衣草的花、叶和茎部分的组合。这些部分可以混合然后捣碎或浸渍,或者将其捣碎或浸渍然后进行组合。然后,可以对得到的经捣碎或经浸渍的花/叶/茎材料使用二氧化碳(CO2作为溶剂进行超临界提取过程。然后,法国薰衣草花/叶/茎的超临界CO2提取物可用于本发明的组合物中。法国薰衣草花/叶/茎的CO2超临界提取物还可以与辛酸/癸酸甘油三酯混合,然后用于本发明的组合物中。在一些情况下,法国薰衣草可商购获得。在一些情况下,法国薰衣草可以以商品名STOCHEY’S由Barnet Products LLC(恩格尔伍德克利夫斯,新泽西(美国))所提供。在本发明的上下文中发现,法国薰衣草提取物可以通过减少或抑制肌管中的钙流入和减少或防止动作电位的产生来减少或抑制肌管收缩。法国薰衣草提取物是有效的局部抗氧化剂,可以减少氧化应激和损害皮肤的自由基。已证明法国薰衣草提取物的应用减少脂质过氧化、增加抗氧化酶活性和降低抗氧化酶活性。
迷迭香叶提取物是来自迷迭香的叶中的提取物。迷迭香原产于地中海地区,是木质多年生草本植物,具有芳香、常绿、针状的叶和白色、粉红色、紫色或蓝色的花。它是高度可以达到最高1.5米的灌木,叶子长度为约2cm至4cm,着色呈绿色(顶面)和白色(底面)。在优选的情况下,迷迭香叶提取物可以从迷迭香的叶中获得。可以使用包含甜菜碱或水合甜菜碱、氢键供体化合物(例如多元醇、有机酸等)和水的流体提取混合物对叶进行共晶提取过程。特别是,可以将叶部分捣碎或浸渍,然后置于上述共晶流体提取混合物以获得共晶提取物。然后,共晶提取物可用于本发明的组合物中。在一些优选的情况下,氢键供体是有机酸,优选乳酸。共晶利用了共晶溶剂,所述共晶溶剂是熔点低于其单独成分熔点的化合物的混合物。在一些情况下,迷迭香可商购获得。在一些情况下,迷迭香可以以商品名RosemaryEUTECTYS BLATM由Naturex(法国)提供。在本发明的上下文中发现,迷迭香叶提取物可以通过减少或抑制肌管中的钙流入和减少或防止动作电位产生,来减少或抑制肌管收缩。还发现了迷迭香叶提取物可以抑制或减少皮肤细胞(例如角质细胞或人真皮成纤维细胞)中MMP-1、MMP-3和MMP-3的产生或活性。迷迭香叶提取物还具有抗氧化和抗炎活性。迷迭香叶提取物改善血液循环,并具有一定的抗菌活性。迷迭香叶提取物包括许多芳香成分,提供了令人愉快的气味。
本文所述的提取物可以是通过本领域已知的提取方法及其组合而制备的提取物。提取方法的非限制性实例包括使用液-液提取、固相提取、水提取、乙酸乙酯、乙醇、丙酮、油、超临界二氧化碳、加热、加压、压降提取、超声提取等。提取物可以是液体、固体、干液、重新悬浮固体等。在某些情况下,每种提取物可以由以下步骤制备:(i)获得期望的植物部分(例如叶、茎、树皮、花、种子等)或整个植物;(ii)捣碎或浸渍植物部分或整个植物;(iii)任选地干燥经捣碎或经浸渍的植物部分或整个植物;(iv)在室温下(例如20℃至30℃)或加热(例如大于30℃或大于30℃,优选30℃至小于溶剂沸点的温度)下,将经捣碎或经浸渍的植物或植物部分置于提取溶剂中足够的时间(例如1分钟、5分钟、10分钟、30分钟、或45分钟或多于45分钟,或1小时、6小时、12小时、或18小时或多于18小时,或2天、3天、4天、5天、6天或多于6天);(v)收集包含提取溶剂和经提取的植物材料(例如,流体提取物)的溶液;和(vi)任选地除去提取溶剂以获得干燥的植物提取物;和(vii)任选地在液体载体(例如水、乙醇、二醇等)中复溶经干燥的植物提取物。在本发明的上下文中还考虑了可以直接使用来自迷迭香叶、法国薰衣草和/或千日菊的植物材料而无需对植物材料进行提取工艺。
C.成分的量
预期本发明的组合物可以包含任意量的本说明书所讨论的成分。该组合物还可以包含任意数目的在本说明书全文中所描述的附加成分的组合(例如,颜料或附加的化妆品或药物成分)。可以改变组合物中成分的浓度。在非限制性实施方案中,例如,千日菊提取物、法国薰衣草提取物和迷迭香叶提取物中的每种在其最终形式中可以独立地包含、基本上组成为或组成为:例如至少大约0.0001%、0.0002%、0.0003%、0.0004%、0.0005%、0.0006%、0.0007%、0.0008%、0.0009%、0.0010%、0.0011%、0.0012%、0.0013%、0.0014%、0.0015%、0.0016%、0.0017%、0.0018%、0.0019%、0.0020%、0.0021%、0.0022%、0.0023%、0.0024%、0.0025%、0.0026%、0.0027%、0.0028%、0.0029%、0.0030%、0.0031%、0.0032%、0.0033%、0.0034%、0.0035%、0.0036%、0.0037%、0.0038%、0.0039%、0.0040%、0.0041%、0.0042%、0.0043%、0.0044%、0.0045%、0.0046%、0.0047%、0.0048%、0.0049%、0.0050%、0.0051%、0.0052%、0.0053%、0.0054%、0.0055%、0.0056%、0.0057%、0.0058%、0.0059%、0.0060%、0.0061%、0.0062%、0.0063%、0.0064%、0.0065%、0.0066%、0.0067%、0.0068%、0.0069%、0.0070%、0.0071%、0.0072%、0.0073%、0.0074%、0.0075%、0.0076%、0.0077%、0.0078%、0.0079%、0.0080%、0.0081%、0.0082%、0.0083%、0.0084%、0.0085%、0.0086%、0.0087%、0.0088%、0.0089%、0.0090%、0.0091%、0.0092%、0.0093%、0.0094%、0.0095%、0.0096%、0.0097%、0.0098%、0.0099%、0.0100%、0.0200%、0.0250%、0.0275%、0.0300%、0.0325%、0.0350%、0.0375%、0.0400%、0.0425%、0.0450%、0.0475%、0.0500%、0.0525%、0.0550%、0.0575%、0.0600%、0.0625%、0.0650%、0.0675%、0.0700%、0.0725%、0.0750%、0.0775%、0.0800%、0.0825%、0.0850%、0.0875%、0.0900%、0.0925%、0.0950%、0.0975%、0.1000%、0.1250%、0.1500%、0.1750%、0.2000%、0.2250%、0.2500%、0.2750%、0.3000%、0.3250%、0.3500%、0.3750%、0.4000%、0.4250%、0.4500%、0.4750%、0.5000%、0.5250%、0.0550%、0.5750%、0.6000%、0.6250%、0.6500%、0.6750%、0.7000%、0.7250%、0.7500%、0.7750%、0.8000%、0.8250%、0.8500%、0.8750%、0.9000%、0.9250%、0.9500%、0.9750%、1.0%、1.1%、1.2%、1.3%、1.4%、1.5%、1.6%、1.7%、1.8%、1.9%、2.0%、2.1%、2.2%、2.3%、2.4%、2.5%、2.6%、2.7%、2.8%、2.9%、3.0%、3.1%、3.2%、3.3%、3.4%、3.5%、3.6%、3.7%、3.8%、3.9%、4.0%、4.1%、4.2%、4.3%、4.4%、4.5%、4.6%、4.7%、4.8%、4.9%、5.0%、5.1%、5.2%、5.3%、5.4%、5.5%、5.6%、5.7%、5.8%、5.9%、6.0%、6.1%、6.2%、6.3%、6.4%、6.5%、6.6%、6.7%、6.8%、6.9%、7.0%、7.1%、7.2%、7.3%、7.4%、7.5%、7.6%、7.7%、7.8%、7.9%、8.0%、8.1%、8.2%、8.3%、8.4%、8.5%、8.6%、8.7%、8.8%、8.9%、9.0%、9.1%、9.2%、9.3%、9.4%、9.5%、9.6%、9.7%、9.8%、9.9%、10%、11%、12%、13%、14%、15%、16%、17%、18%、19%、20%、21%、22%、23%、24%、25%、26%、27%、28%、29%、30%、35%、40%、45%、50%、60%、65%、70%、75%、80%、85%、90%、95%或99%或其中可获得的任何范围的在本说明书的全文和权利要求所提及的至少一种成分。在非限制性方面,该百分比可以按整个组合物的重量或体积进行计算。本领域普通技术人员会理解,给定组合物中的浓度可以根据成分的添加、替换和/或减少而改变。
D.载剂
本发明的组合物可以包含所有类型的载剂和载体,或掺入到所有类型的载剂和载体中。载体或载剂可以是药学上或皮肤病学上可接受的载体或载剂。载体和载剂的非限制性实例包括水、甘油、醇、油、含硅化合物、含硅酮的化合物和蜡。变体和其它合适的载体对于熟练技术人员是明显的,并且适用于本发明。在某些方面,化合物、成分和试剂的浓度和组合以这样的方式进行选择,使得组合是化学相容的并且不形成从最终产品中沉淀出来的复合物。
E.结构
本发明的组合物可以被构建或配制为多种不同的形式。非限制性实例包括乳液(例如,油包水、水包油包水、水包油、水包硅酮、硅酮包水、油包水包油、硅酮包水包油的乳液)、霜剂、爽肤水、溶液(水溶液或者水-乙醇溶液)、无水基质(例如口红和粉剂)、凝胶、面膜、磨砂膏、身体乳、peels和软膏。变体和其它的结构对于熟练技术人员是明显的,并且适用于本发明。
F.附加成分
除了本发明所公开成分的组合之外,组合物还可以包含附加成分,例如化妆品成分和药物有效成分。这些附加成分的非限制性实例在以下子章节中进行描述。
1、化妆品成分
CTFA国际化妆品成分词典和手册(2004和2008)描述了多种可以在本发明的环境下使用的非限制性化妆品成分。这些成分种类的实例包括:芳香剂(人造的和天然的;例如葡萄糖酸、苯氧乙醇和三乙醇胺)、染料和着色成分(例如蓝1、蓝1色淀、红40、二氧化钛、D&C蓝色4号、D&C绿色5号、D&C橙色4号、D&C红色17号、D&C红色33号、D&C紫色2号、D&C黄色10号和D&C黄色11号)、调味剂/香味剂(例如,甜菊(甜叶)提取物和薄荷醇)、吸附剂、润滑剂、溶剂、保湿剂(包括例如润肤剂、湿润剂、成膜剂、闭塞剂和影响皮肤天然保湿机制的试剂)、拒水剂、UV吸收剂(物理和化学吸收剂,例如对氨基苯甲酸(“PABA”)和相应的PABA衍生物、二氧化钛、氧化锌等)、精油、维生素(例如B、D、E和K)、微量金属(例如锌、钙和硒)、抗刺激剂(例如类固醇和非-类固醇抗炎药)、植物提取物(例如芦荟、柑橘、黄瓜提取物、银杏、人参和迷迭香)、抗菌剂、抗氧化剂(例如BHT和生育酚)、螯合剂(例如EDTA二钠和EDTA四钠)、防腐剂(例如对羟基苯甲酸甲酯和对羟基苯甲酸丙酯)、pH调节剂(例如氢氧化钠和柠檬酸)、吸收剂(例如淀粉辛烯基琥珀酸铝、高岭土、玉米淀粉、燕麦淀粉、环糊精、滑石和沸石)、皮肤漂白和亮肤剂(例如氢醌和乳酸烟酰胺)、湿润剂(例如山梨醇、脲、甲基葡糖醇聚醚-20、糖类同分异构体和甘露醇)、去角质剂、防水剂(例如氢氧化镁/铝硬脂酸盐)、皮肤调节剂(例如芦荟提取物、尿囊素、没药醇、神经酰胺、聚二甲基硅氧烷、透明质酸、生物糖胶-1、乙基己基甘油、戊二醇、氢化聚癸烯、辛基十二烷基油酸酯和甘草酸二钾)。这些成分的一些非限制性实例在以下子章节中提供。
a.UV吸收剂和/或反射剂
可以与本发明的组合物组合使用的UV吸收剂和/或反射剂包括化学和物理防晒物质。可以使用的化学防晒物质的非限制性实例包括对氨基苯甲酸(PABA)、PABA酯(PABA甘油酯、PABA戊基二甲酯和PABA辛基二甲酯)、PABA丁酯、PABA乙酯、PABA乙基二羟基丙酯、二苯甲酮(羟苯甲酮、磺异苯酮、二苯甲酮和二苯甲酮-1至二苯甲酮-12)、肉桂酸酯(甲氧基肉桂酸辛酯(桂皮酸酯))、对-甲氧基肉桂酸异戊酯、肉桂酸辛基甲氧基酯、西诺沙酯、肉桂酸二异丙基甲酯、甲氧基肉桂酸DEA酯、二异丙基肉桂酸乙酯、二甲氧基肉桂酸甘油辛酸酯和甲氧基肉桂酸乙酯)、肉桂酸酯、水杨酸酯(均甲基水杨酸酯、水杨酸苄酯、水杨酸乙二醇酯、水杨酸异丙苄酯等)、邻氨基苯甲酸酯、尿刊酸乙酯、胡莫柳酯、水杨酸辛酯、二苯甲酰基甲烷衍生物(例如阿伏苯宗)、奥克立林、辛基三嗪酮、棓酰棓酸三油酸酯、氨基苯甲酸甘油酯、2-羟基-1,4-萘醌与二羟基丙酮、乙基己基三嗪酮、二辛基丁酰胺基三嗪酮、苯亚甲基丙二酸脂聚硅氧烷、对苯二亚甲基二樟脑磺酸、苯基二苯并咪唑四磺酸酯二钠、二乙氨基羟基苯甲酰基苯甲酸己酯、双二乙氨基羟基苯甲酰基苯甲酸酯、双苯并唑基苯基乙基己基亚氨基三嗪、甲酚曲唑三硅氧烷、亚甲基双苯并三唑基四甲基丁基苯酚和双乙基己基氧苯酚甲氧苯基三嗪、4-甲基苯亚甲基樟脑和4-甲氧基肉桂酸异戊酯。物理防晒物质的非限制性实例包括高岭土、滑石、矿脂和金属氧化物(例如二氧化钛和氧化锌)。
b.保湿剂
可以与本发明的组合物一起使用的保湿剂的非限制性实例包括氨基酸、硫酸软骨素、双甘油、赤藓糖醇、果糖、葡萄糖、甘油、甘油聚合物、乙二醇、1,2,6-己三醇、蜂蜜、透明质酸、氢化蜂蜜、氢化淀粉水解物、肌醇、乳糖醇、麦芽糖醇、麦芽糖、甘露醇、天然保湿因子、PEG-15丁二醇、聚甘油山梨醇、吡咯烷酮羧酸的盐、PCA钾、丙二醇、糖类同分异构体、葡糖醛酸钠、PCA钠、山梨醇、蔗糖、海藻糖、脲和木糖醇。
其他实例包括乙酰化羊毛脂、乙酰化羊毛脂醇、丙氨酸、藻类提取物、库拉索芦荟(Aloe barbadensis)、库拉索芦荟提取物、库拉索芦荟凝胶、药蜀葵(althea officinalis)提取物、杏(Prunus armeniaca)仁油、精氨酸、精氨酸天冬氨酸酯、山金车(Arnicamontana)提取物、天冬氨酸、鳄梨(Persea gratissima)油、屏障鞘脂、丁醇、蜂蜡、山萮醇、β-谷甾醇、白桦(Betula alba)树皮提取物、琉璃苣(Borago officinalis)提取物、假叶树(Ruscus aculeatus)提取物、丁二醇、金盏花(Calendula officinalis)提取物、金盏花油、小烛树(Euphorbia cerifera)蜡、菜籽油、辛酸/癸酸甘油三酯、小豆蔻(Elettariacardamomum)油、巴西棕榈(Copernicia erifera)蜡、胡萝卜(Daucus carota sativa)油、蓖麻(Ricinus communis)油、神经酰胺、地蜡、鲸蜡硬脂醇聚醚-5、鲸蜡硬脂醇聚醚-12、鲸蜡硬脂醇聚醚-20、鲸蜡硬脂醇辛酸酯、鲸蜡醇聚醚-20、鲸蜡醇聚醚-24、鲸蜡醇乙酸酯、鲸蜡醇辛酸酯、鲸蜡醇棕榈酸酯、洋甘菊(Anthemis nobilis)油、胆固醇、胆固醇酯、胆甾醇羟基硬脂酸酯、柠檬酸、鼠尾草(Salvia sclarea)油、可可(Theobroma cacao)脂、椰油醇-辛酸酯/癸酸酯、椰子(Cocos nucifera)油、胶原蛋白、胶原蛋白氨基酸、玉米(Zea mays)油、脂肪酸、油酸癸酯、聚二甲基硅氧烷共聚醇、聚二甲基硅氧烷醇、己二酸二辛酯、琥珀酸二辛酯、二聚季戊四醇六辛酸酯/六癸酸酯、DNA、赤藓糖醇、乙氧基二乙二醇、亚油酸乙酯、蓝桉(Eucalyptus globulus)油、月见草(Oenothera biennis)油、脂肪酸、斑点老鹳草(Geranium maculatum)油、葡萄糖胺、葡糖谷氨酸酯、谷氨酸、甘油聚醚-26、甘油、丙三醇、二硬脂酸甘油酯、羟基硬脂酸甘油酯、月桂酸甘油酯、亚油酸甘油酯、豆蔻酸甘油酯、油酸甘油酯、硬脂酸甘油酯、硬脂酸甘油酯SE、甘氨酸、乙二醇硬酯酸酯、乙二醇硬酯酸酯SE、葡糖氨基葡聚糖、葡萄(Vitis vinifera)籽油、美洲榛(Corylus americana)坚果油、欧洲榛(Corylus americana)坚果油、己二醇、透明质酸、红花(Carthamus tinctorius)油、氢化蓖麻油、氢化椰油甘油酯、氢化椰子油、氢化羊毛脂、氢化卵磷脂、氢化棕榈油甘油酯、氢化棕榈仁油、氢化大豆油、氢化牛脂酸甘油酯、氢化植物油、水解胶原蛋白、水解弹性蛋白、水解葡糖氨基葡聚糖、水解角蛋白、水解大豆蛋白、羟基化羊毛脂、羟基脯氨酸、异鲸蜡醇硬脂酸酯、异鲸蜡醇硬脂酰氧基硬脂酸酯、油酸异癸酯、异硬脂酸异丙酯、羊毛脂酸异丙酯、肉豆蔻酸异丙酯、棕榈酸异丙酯、硬脂酸异丙酯、异硬脂酰胺DEA、异硬脂酸、异硬脂醇乳酸酯、异硬脂醇新戊酸酯、茉莉(Jasminum officinale)油、霍霍巴(Buxus chinensis)油、巨藻、石栗(Aleurites moluccana)坚果油、乳酰胺MEA、羊毛脂醇聚醚-16、羊毛脂醇聚醚-10乙酸酯、羊毛脂、羊毛脂酸、羊毛脂醇、羊毛脂油、羊毛脂蜡、薰衣草(Lavandula angustifolia)油、卵磷脂、柠檬(Citrus medica limonum)油、亚油酸、亚麻酸、澳洲坚果(Macadamiaternifolia)油、麦芽糖醇、母菊(Chamomilla recutita)油、甲基葡糖倍半硬脂酸酯、甲基硅烷醇PCA酯、矿物油、貂油、黄褐色被孢霉油、肉豆蔻醇乳酸酯、肉豆蔻醇肉豆蔻酸酯、肉豆蔻醇丙酸酯、新戊二醇二辛酸酯/二癸酸酯、辛基十二醇、肉豆蔻酸辛基十二醇酯、硬脂酰氧基硬脂酸辛基十二醇酯、羟基硬脂酸辛酯、棕榈酸辛酯、水杨酸辛酯、硬脂酸辛酯、油酸、橄榄(Olea europaea)油、橙(Citrus aurantium dulcis)油、棕榈(Elaeis guineensis)油、棕榈酸、泛硫乙胺、泛醇、泛醇基乙基醚、石蜡、PCA、桃(Prunus persica)仁油、花生(Arachis hypogaea)油、PEG-8C12-18酸酯、PEG-15椰油胺、PEG-150二硬脂酸酯、PEG-60甘油异硬脂酸酯、PEG-5甘油硬脂酸酯、PEG-30甘油硬脂酸酯、PEG-7氢化蓖麻油、PEG-40氢化蓖麻油、PEG-60氢化蓖麻油、PEG-20甲基葡糖倍半硬脂酸酯、PEG-40失水山梨醇全油酸酯、PEG-5大豆甾醇、PEG-10大豆甾醇、PEG-2硬脂酸酯、PEG-8硬脂酸酯、PEG-20硬脂酸酯、PEG-32硬脂酸酯、PEG-40硬脂酸酯、PEG-50硬脂酸酯、PEG-100硬脂酸酯、PEG-150硬脂酸酯、十五内酯、薄荷(Mentha piperita)油、矿脂、磷脂、浮游生物提取物、多氨基酸多糖缩合物、聚甘油-3二异硬脂酸酯、聚季铵盐-24、聚山梨醇酯-20、聚山梨醇酯-40、聚山梨醇酯-60、聚山梨醇酯-80、聚山梨醇酯-85、肉豆蔻酸钾、棕榈酸钾、丙二醇、丙二醇二辛酸酯/二癸酸酯、丙二醇二辛酸酯、丙二醇二壬酸酯、丙二醇月桂酸酯、丙二醇硬脂酸酯、丙二醇硬脂酸酯SE、PVP、吡哆醇二棕榈酸酯、视黄醇、视黄醇棕榈酸酯、米(Oryza sativa)糠油、RNA、迷迭香(Rosmarinus officinalis)油、玫瑰油、红花(Carthamus tinctorius)油、鼠尾草(Salviaofficinalis)油、檀香(Santalum album)油、丝氨酸、血清蛋白、芝麻(Sesamumindicum)油、牛油果(Butyrospermum parkii)脂、蚕丝粉、软骨素硫酸钠、透明质酸钠、乳酸钠、棕榈酸钠、PCA钠、聚谷氨酸钠、可溶性胶原、失水山梨醇月桂酸酯、失水山梨醇油酸酯、失水山梨醇棕榈酸酯、失水山梨醇倍半油酸酯、失水山梨醇硬脂酸酯、山梨醇、大豆(Glycine soja)油、鞘脂、角鲨烷、角鲨烯、硬脂酰胺MEA-硬脂酸酯、硬脂酸、硬脂氧基聚二甲基硅氧烷、硬脂氧基三甲基硅烷、硬脂醇、硬脂醇甘草亭酸酯、硬脂醇庚酸酯、硬脂醇硬脂酸酯、向日葵(Helianthus annuus)籽油、甜扁桃仁(Prunus amygdalus dulcis)油、合成蜂蜡、生育酚、生育酚乙酸酯、生育酚亚油酸酯、三山嵛精、十三烷醇新戊酸酯、十三烷醇硬脂酸酯、三乙醇胺、三硬脂精、脲、植物油、水、蜡、小麦(Triticum vulgare)胚芽油和依兰(Canangaodorata)油。
c.抗氧化剂
额外的抗氧化剂可以与法国薰衣草提取物和迷迭香叶提取物结合使用。可以使用的抗氧化剂的非限制性实例包括乙酰半胱氨酸、抗坏血酸衍生物如抗坏血酸多肽、抗坏血酸二棕榈酸酯、抗坏血酸甲基硅烷醇果胶酸酯、抗坏血酸棕榈酸酯、抗坏血酸硬脂酸酯、BHA、BHT、叔丁基氢醌、半胱氨酸、半胱氨酸HCI、二戊基氢醌、二叔丁基氢醌、二鲸蜡醇硫代二丙酸酯、二油基生育酚甲基硅烷醇、抗坏血酸硫酸酯二钠、二硬脂醇硫代二丙酸酯、双十三烷醇硫代二丙酸酯、没食子酸月桂酯、异抗坏血酸、抗坏血酸酯、阿魏酸乙酯、阿魏酸、没食子酸酯、氢醌、巯基乙酸异辛酯、曲酸、抗坏血酸镁、抗坏血酸磷酸酯镁、甲基硅烷醇抗坏血酸酯、天然植物抗氧化剂例如绿茶或葡萄籽提取物、去甲二氢愈创木酸、没食子酸辛酯、苯巯基乙酸、磷酸抗坏血酸酯生育酚酯钾、亚硫酸钾、没食子酸丙酯、醌、迷迭香酸、抗坏血酸钠、亚硫酸氢钠、异抗坏血酸钠、焦亚硫酸钠、亚硫酸钠、超氧化物歧化酶、巯基乙酸钠、山梨醇缩糠醛、硫二甘醇、亚硫基二乙酰胺、硫代二乙酸、巯基乙酸、硫代乳酸、硫代水杨酸、生育酚聚醚-5、生育酚聚醚-10、生育酚聚醚-12、生育酚聚醚-18、生育酚聚醚-50、生育酚、托可索伦、生育酚乙酸酯、生育酚亚油酸酯、生育酚烟酸酯、生育酚琥珀酸酯和三(壬基酚)亚磷酸酯。
d.结构化剂
在其它非限制性方面,本发明的组合物可以包含结构化剂。在某些方面,结构化剂帮助向组合物提供流变学特征以有助于组合物的稳定性。在其它方面,结构化剂还可以起乳化剂或表面活性剂的作用。结构化剂的非限制性实例包括硬脂酸、棕榈酸、硬脂醇、鲸蜡醇、山嵛醇、硬脂酸、棕榈酸、具有平均约1个至约21个环氧乙烷单元的硬脂醇的聚乙二醇醚、具有平均约1个至约5个环氧乙烷单元的鲸蜡醇的聚乙二醇醚及其混合物。
e.乳化剂
乳化剂可以降低相间界面张力并改善乳液的剂型和稳定性。乳化剂可以是非离子的、阳离子的、阴离子的和两性离子的乳化剂(参见McCutcheon's(1986);美国专利第5,011,681号;第4,421,769号;第3,755,560号)。非限制性实例包括甘油酯、丙二醇酯、丙二醇的脂肪酸酯、聚丙二醇的脂肪酸酯、山梨醇的酯、失水山梨醇酐酯、羧酸共聚物、葡萄糖的酯和醚、乙氧基化的酯、乙氧基化的醇、磷酸烷基酯、聚氧乙烯脂肪醚磷酸酯、脂肪酸酰胺、乳酰乳酸酯、脂肪酸盐、TEA硬脂酸酯、DEA油醇聚氧乙烯醚-3磷酸酯、聚乙二醇20、失水山梨醇单月桂酸酯(聚山梨醇酯20)、聚乙二醇5大豆甾醇、硬脂醇聚氧乙烯醚-2、硬脂醇聚氧乙烯醚-20、硬脂醇聚氧乙烯醚-21、鲸蜡硬脂醇聚醚-20、鲸蜡硬脂基葡糖苷、鲸蜡硬脂醇、C12-13链烷醇聚醚-3、PPG-2甲基葡糖醚二硬脂酸酯、PPG-5鲸蜡基醚-20、双-PEG/PPG-20/20聚二甲基硅氧烷、鲸蜡醇聚醚-10、聚山梨醇酯80、鲸蜡醇磷酸酯、鲸蜡醇磷酸酯钾、二乙醇胺鲸蜡醇磷酸酯、聚山梨醇酯60、甘油硬脂酸酯、PEG-100硬脂酸酯、花生醇、花生醇葡糖苷及其混合物。
f.含有硅酮的化合物
在非限制性方面,含硅酮的化合物包括分子主链由交替的硅和氧原子与连接在硅原子上的侧基组成的聚合产物家族中的任何成员。通过改变-Si-O-链的长度、侧基和交联,硅酮可以合成为各种各样的材料。它们的稠度可以从液体到凝胶到固体改变。
可以在本发明的环境下使用的含硅酮的化合物包括本说明书中所描述的或本领域普通技术人员已知的那些。非限制性实例包含硅油(例如挥发性和非挥发性油)、凝胶和固体。在一些方面,含硅化合物包括硅油,例如聚有机硅氧烷。聚有机硅氧烷的非限制性实例包括聚二甲基硅氧烷、环聚二甲基硅氧烷、聚硅氧烷-11、苯基聚三甲基硅氧烷、三甲基甲硅烷氧基氨端聚二甲基硅氧烷(trimethylsilylamodimethicone)、硬脂氧基三甲基硅烷或其混合物和其它任何给定比例的有机硅氧烷材料,以根据预期的应用(例如,对特定区域例如皮肤、毛发或眼睛)达到期望的稠度和应用特征。“挥发性硅油”包括具有低气化热的硅油,即通常低于约50卡每克硅油。挥发性硅油的非限制性实例包括:环聚二甲基硅氧烷,例如Dow Corning344Fluid、Dow Corning 345Fluid、Dow Corning 244Fluid和Dow Corning245Fluid、Volatile Silicon 7207(康涅狄格州丹伯里的Union Carbide Corp.);低黏度聚二甲基硅氧烷,即黏度为约50cst或低于50cst的聚二甲基硅氧烷(例如聚二甲基硅氧烷,例如Dow Corning 200-0.5cst Fluid)。Dow Corning Fluid可以从密歇根州米德兰的DowCorning Corporation购得。在CTFA化妆品成分词典的第三版中(通过引用并入),环聚二甲基硅氧烷和聚二甲基硅氧烷分别被描述为环状二甲基聚硅氧烷化合物和用三甲基硅氧基单元封端的完全甲基化的线性硅氧烷的混合物。可以在本发明的环境下使用的其它非限制性挥发性硅油包括从纽约州沃特福德的General Electric Co.、Silicone Products Div.和密歇根州艾德里安的Stauffer Chemical Co.的SWS Silicones Div.购得的那些。
g.去角质剂
去角质剂包括除去皮肤外表面上的死皮肤细胞的成分。这些试剂可以通过机械的、化学的和/或其他的方式起作用。机械去角质剂的非限制性实例包括磨料,例如浮石、二氧化硅、织物、纸、壳、珠、固体晶体、固体聚合物等。化学去角质剂的非限制性实例包括酸和酶去角质剂。可用作去角质剂的酸包括但不限于乙醇酸、乳酸、柠檬酸、α羟基酸、β羟基酸等。本领域技术人员已知的其他去角质剂也也预期在本发明的上下文中有用。
h.精油
精油包括来自药草、花朵、树木和其它植物的油。这些油一般以植物细胞间微小的液滴存在,并可以通过本领域技术人员已知的若干方法(例如,蒸气蒸馏、脂吸法(即通过使用脂肪提取)、浸渍、溶剂提取或机械压榨)进行提取。当这些类型的油暴露于空气时,其趋于挥发(即挥发性油)。因此,虽然许多精油是无色的,但是随着时间其被氧化并且颜色变得更深。精油不溶于水,但溶于醇、醚、固定油(植物的)和其它有机溶剂。在精油中发现的一般物理特征包括约160℃至240℃的沸点和约0.759至约1.096的密度。
精油一般通过发现油的来源植物命名。例如,玫瑰油或薄荷油分别来自玫瑰或薄荷植物。可以在本发明的环境下使用的精油的非限制性实例包括芝麻油、澳洲坚果油、茶树油、月见草油、西班牙鼠尾草油、西班牙迷迭香油、芫荽油、百里香油、麝香草油、玫瑰油、大茴香油、凤仙花油、香柠檬油、玫瑰木油、香柏油、甘菊油、鼠尾草油、香紫苏油、丁香油、柏木油、桉油、茴香油、海茴香油、乳香油、香叶油、姜油、葡萄柚油、茉莉油、杜松子油、薰衣草油、柠檬油、柠檬草油、梨莓油、来檬油、马郁兰油、没药油、苦橙花油、橙油、绿叶油、胡椒油、黑胡椒油、苦橙叶油、松油、奥图玫瑰油、迷迭香油、檀香油、绿薄荷油、甘松油、香根草油、冬青油、依兰油。还预期本领域技术人员已知的其它精油在本发明的环境下是有用的。
i.增稠剂
包括增稠剂或凝胶剂在内的增稠剂,包括可以增加组合物黏度的物质。增稠剂包括可以增加组合物黏度而基本上不改变组合物内活性成分功效的那些。增稠剂还可以增加本发明的组合物的稳定性。在本发明的一些方面,增稠剂包括氢化聚异丁烯、三羟基硬脂酸甘油酯、丙烯酰二甲基牛磺酸铵/vp共聚物、丙烯酸酯类/C10-30烷醇丙烯酸酯交联聚合物或其混合物。
可以在本发明的环境下使用的另外的增稠剂的非限制性实例包括羧酸聚合物、交联的聚丙烯酸酯聚合物、聚丙烯酰胺聚合物、多糖和胶。羧酸聚合物的实例包括含有一种或多于一种衍生自丙烯酸的单体的交联化合物、取代的丙烯酸和这些丙烯酸和取代的丙烯酸的盐和酯,其中所述交联剂含有两个或多于两个碳-碳双键,并衍生自多元醇(参见美国专利第5087445号;第4509949号;第2798053号;CTFAInternational Cosmetic IngredientDictionary,Fourth edition,1991,pp.12and 80)。市售羧酸聚合物的实例包括卡波姆,其为丙烯酸与蔗糖或季戊四醇的烯丙基醚交联的均聚物(例如,购自B.F.Goodrich的CARBOPOLTM900系列)。
交联的聚丙烯酸酯聚合物的非限制性实例包括阳离子型和非离子型聚合物。实例描述于美国专利第5100660号、第4849484号、第4835206号、第4628078号、第4599379号。
聚丙烯酰胺聚合物(包括非离子的聚丙烯酰胺聚合物,其包含经取代的支化或未支化的聚合物)的非限制性实例包括聚丙烯酰胺、异构烷烃和月桂醇聚氧醚-7、丙烯酰胺和经取代的丙烯酰胺与丙烯酸和经取代的丙烯酸的多嵌段共聚物。
多糖的非限制性实例包括纤维素、羧甲基羟乙基纤维素、乙酸丙酸羧酸纤维素、羟乙基纤维素、羟乙基乙基纤维素、羟丙基纤维素、羟丙基甲基纤维素、甲基羟乙基纤维素、微晶纤维素、纤维素硫酸钠及其混合物。其他实例为烷基取代的纤维素,其中纤维素聚合物的羟基被羟烷基化(优选地羟乙基化或羟丙基化)以形成羟烷基化的纤维素,其然后用C10至C30直链或带支链的烷基基团通过醚键进行进一步改性。一般这些聚合物为C10至C30直链或带支链的醇与羟烷基纤维素的醚。其它有用的多糖包括硬葡聚糖类,其包含每三个单元具有一个(1-6)连接的葡萄糖的(1-3)连接的葡萄糖单元的直链。
本发明可以使用的胶的非限制性实例包括阿拉伯树胶、琼脂、藻胶、藻酸、藻酸铵、支链淀粉、藻酸钙、角叉菜胶钙、肉毒碱、角叉菜胶、糊精、明胶、结冷胶、瓜尔豆胶、瓜尔胶羟丙基三甲基氯化铵、锂蒙脱石、透明质酸、水合二氧化硅、羟丙基壳聚糖、羟丙基瓜尔胶、卡拉亚胶、巨藻、角豆胶、纳豆胶、藻酸钾、角叉菜胶钾、藻酸丙二醇酯、菌核胶、羧甲基葡聚糖钠、角叉菜胶钠、黄蓍胶、黄原胶及其混合物。
j.防腐剂
可以在本发明的环境下使用的防腐剂的非限制性实例包括季铵盐防腐剂如聚季铵盐-1和苄烷铵卤化物(例如苯扎氯铵(“BAC”)和苯扎溴铵))、对羟基苯甲酸酯(例如对羟基苯甲酸甲酯和对羟基苯甲酸丙酯)、苯氧基乙醇、苄醇、氯丁醇、苯酚、山梨酸、硫柳汞或其组合。
k.润肤剂
有用的润肤剂包括以下:(a)硅油及其修饰物,例如线性和环状聚二甲基硅氧烷;氨基、烷基、烷芳基和芳基硅油;(b)脂肪和油,包括天然脂肪和油例如荷荷巴油、大豆油、葵花油、米糠油、鳄梨油、杏仁油、橄榄油、芝麻油、桃仁油、蓖麻油、椰子油、貂油;可可脂肪油;牛油、猪油;通过使上述油氢化获得的氢化油;以及合成单甘油酯、甘油二酯和甘油三酯,例如肉豆蔻酸甘油酯和2-乙基己酸甘油酯;(c)蜡,例如巴西蜡棕榈、鲸蜡、蜂蜡、羊毛脂及其衍生物;(d)疏水性植物提取物;(e)烃类,例如液状石蜡、凡士林、微晶蜡、纯地蜡、鲨烯、姥鲛烷和矿物油;(f)高级脂肪酸,例如月桂酸、肉豆蔻酸、棕榈酸、硬脂酸、山萮酸、亚油酸、亚麻油酸、亚麻酸、羊毛酸、异硬脂酸、花生四烯酸和多不饱和脂肪酸(PUFA);(g)高级醇,例如月桂醇、鲸蜡醇、硬脂醇、油醇、二十二醇、胆固醇和2-己基癸醇;(h)酯,例如辛酸十六酯、合成油脂剂、十六烷基乳酸酯、肉豆蔻酸异丙酯、肉豆蔻醇肉豆蔻酸酯、棕榈酸异丙酯、硬脂酸丁酯、油酸癸酯、异硬脂酸酯、单硬脂酸甘油酯、甘油二硬脂酸酯、甘油三硬脂酸酯、烷醇乳酸酯、柠檬酸酯和醇酒石酸酯;(i)精油及其提取物,例如薄荷提取物、茉莉提取物、樟脑提取物、白杉木提取物、苦橙皮提取物、ryu提取物、松节油、肉桂提取物、佛手柑提取物、蜜柑提取物、菖蒲提取物、松树提取物、薰衣草提取物、月桂提取物、丁香提取物、罗汉柏提取物、桉树提取物、柠檬提取物、七瓣莲提取物、百里香提取物、椒样薄荷提取物、玫瑰精油、鼠尾草提取物、芝麻油、姜油、罗勒提取物、杜松提取物、柠檬草提取物、迷迭香提取物、紫檀提取物、鳄梨提取物、葡萄提取物、葡萄籽油、没药提取物、黄瓜提取物、豆瓣菜提取物、金盏花提取物、接骨木花提取物、天竺葵提取物、菩提花提取物、苋菜提取物、海藻提取物、银杏提取物、西洋参提取物、胡萝卜提取物、瓜拉那提取物、茶树精油、荷荷巴油、紫草提取物、燕麦提取物、可可提取物、橙花精油、香草提取物、绿茶提取物、薄荷油、芦荟提取物、薄荷醇、桉叶素、丁香酚、柠檬醛、香茅提取物、冰片提取物、芳樟醇、香叶醇、月见草提取物、樟脑、麝香草酚、盘龙参酚(Spirantol)、蒎烯(penene)、柠檬烯和萜油;(j)脂类,例如胆固醇、神经酰胺、蔗糖酯和假神经酰胺,如欧洲专利说明书第556957号中所描述的;(k)维生素、矿物质和皮肤营养物,例如维生素A、维生素E和维生素K;维生素烷基酯,包括维生素C烷基酯;镁、钙和牛奶;(l)防晒霜,例如辛基甲氧基肉桂酸(Parsol MCX)和丁基甲氧基苯甲酰甲烷(Parsol1789);(l)磷脂;(m)多羟基醇,例如甘油和丙二醇;和多元醇,例如聚乙二醇;(n)抗老化组合物,例如α-羟基酸、β-羟基酸;和(o)前述任意一种组分的混合物等。
l.增黏剂
合适的增黏剂的实例包括但不限于脂肪族烃树脂、芳香族改性脂肪烃树脂、氢化聚环戊二烯树脂、环戊二烯树脂、松香、松香酯、木松香、木松香酯、塔罗油松香、塔罗油松香酯、多萜、芳香族改性多萜、萜烯酚、芳香改性氢化聚环戊二烯树脂、氢化脂肪族树脂、氢化脂肪族芳香树脂、氢化萜烯和改性的萜烯、氢化松香酸、氢化松香酯、聚异戊二烯、部分或完全氢化的聚异戊二烯、聚丁二烯、部分或完全氢化的聚丁二烯等。如一些引用的实例所证明的,增黏剂可以是完全或部分氢化的。增黏剂也可以是非极性的。(非极性表示增黏剂基本上不含具有极性基团的单体。优选地,极性基团不存在,然而,如果它们存在,优选以最高为约5重量%,优选最高为约2重量%,更优选最高为约0.5重量%的量存在。)。
m.着色剂
本发明的组合物还含有至少一种化妆品可接受的着色剂,例如颜料和染料。合适的颜料的实例包括但不限于无机颜料、有机颜料、色淀、珠光颜料、虹彩或光学可变颜料及其混合物。颜料应理解为表示无机或有机、白色或有色颗的颗粒。在本发明的范围内,所述颜料可以任选地进行表面处理,但不限于例如硅酮、全氟化合物、卵磷脂和氨基酸的处理。
n.表面活性剂
在本发明的组合物中作为表面活性剂组分的有用的表面活性剂包括非离子、阴离子、阳离子和两性(两性离子)表面活性剂,并可以互相组合使用。
o.pH调节剂
pH调节剂包括无机和有机酸和碱,特别是氨水、柠檬酸、磷酸、乙酸、氢氧化钠、乳酸、乙酰丙酸、乙醇酸、酒石酸、苹果酸、吡咯烷酮羧酸(PCA)、琥珀酸、柠檬酸、谷氨酸、2-氨基-2-甲基-1-丙醇(AMP)和三乙醇胺(TEA)。
p.还原剂
合适的还原剂包括但不限于硫脲、硫代硫酸盐(如钠盐)、亚硫酸盐、亚硫酸氢盐、偏亚硫酸氢盐、硼氢化物和次磷酸盐、抗坏血酸及其盐、酯和衍生物(如棕榈酸抗坏血酸酯和抗坏血酸多肽)、以及生育酚及其盐、酯及其衍生物(如乙酸生育酚酯)。其他还原剂列于INCI手册的第1655-56页。
q.芳香剂
本文公开的组合物可以任选地包含芳香剂。可能的芳香剂的实例包括天然油或天然衍生材料,以及合成芳香剂,例如烃、醇、醛、酮、酯、内酯、醚、腈和多官能团化合物。天然油的非限制性实例包括以下物质:罗勒(Ocimum basilicum)油、月桂(Pimento acris)油、香蜂草(Monarda didyma)油、香柠檬(Citrus aurantium bergamia)油、小豆蔻(Elettariacardamomum)油、大西洋雪松(Cedrus atlantica)油、洋甘菊(Anthemis nobilis)油、肉桂(Cinnamomum cassia)油、香茅(Cymbopogon nardus)油、鼠尾草(Salvia sclarea)油、丁香(Eugenia caryophyllus)油、丁香叶(Eufenia caryophyllus)油、油莎豆油、柏树(Cupressus sempervirens)油、柠檬桉油、斑点老鹳草油、生姜(Zingiber officinale)油、葡萄柚(Citrus grandis)油、榛子(Corylus avellana)果油、素馨(Jasminum officinale)油、杜松子油、杜松焦油、北美圆柏油、奇异果(Actinidia chinensis)水、杂熏衣草(Lavandula hybrida)油、薰衣草(Lavandula angustifolia)油、薰衣草(Lavandulaangustifolia)水、柠檬(Citrus medica limonum)油、柠檬草(Cymbopogon schoenanthus)油、酸橙(Citrus aurantifolia)油、椴树(Tilia cordata)油、椴树(Tilia cordata)水、蜜橘(Citrus nobilis)油、肉豆蔻(Myristica fragrans)油、柑橘(Citrus aurantiumdulcis)花油、柑橘(Citrus aurantium dulcis)油、柑橘(Citrus aurantium dulcis)水、广藿香(Pogostemon cablin)油、薄荷(Menthe piperita)油、薄荷(Menthe peperita)水、迷迭香(Rosmarinus officinalis)油、玫瑰油、玫瑰(Rosa damascena)提取物、玫瑰(Rosamultiflora)提取物、紫檀(Aniba rosaeodora)提取物、鼠尾草(Salvia officinalis)油、檀香(Santalum album)油、留兰香(Menthe viridis)油、茶树(Melaleuca alternifolia)油、薄荷醇和依兰依兰(Cananga odorata)油。合成烃芳香剂的一些非限制性实例包括石竹烯、β-法尼烯、柠檬烯、α-蒎烯和β-蒎烯。合成醇芳香剂的一些非限制性实例包括檀香、香茅醇、芳樟醇、苯乙醇和α-松油醇(R═H)。合成醛芳香剂的一些非限制性实例包括2-甲基十一碳烯醛、柠檬醛、己基肉桂醛、异环柠檬醛、铃兰醛和10-十一碳烯醛。合成酮芳香剂的一些非限制性实例包括开司米酮、α-紫罗酮、龙涎酮E、乙酰基二聚异常戊烯、麝香酮和吐纳麝香。合成酯芳香剂的一些非限制性实例包括乙酸苄酯、4-叔丁基环己基乙酸酯(顺式和反式)、乙酸柏木酯、乙酸三环癸烯酯、醋酸异冰片酯和α-乙酸松油酯(R=乙酰基)。合成内酯芳香剂的一些非限制性实例包括香豆素、茉莉内酯、环十五内酯(muskalactone)和桃醛。合成醚芳香剂的一些非限制性实例包括降龙涎香醚、花药和佳乐麝香。合成腈芳香剂的一些非限制性实例包括肉桂腈和gernonitrile。最后,合成多官能化合物的芳香剂的一些非限制性实例包括水杨酸戊酯、异丁香酚、二氢茉莉酮酸甲酯、天芥菜精、新铃兰醛和香草醛。
r.发泡剂
发泡剂包括例如月桂基硫酸钠、月桂酰基肌氨酸钠、烷基磺基琥珀酸钠、椰子油脂肪酸单甘醇磺酸钠、α-烯基磺酸钠、N-酰氨基酸盐如N-脂肪酰基谷氨酸盐、2-烷基-N-羧甲基-N-羟乙基咪唑啉甜菜碱、麦芽糖醇脂肪酸酯、蔗糖脂肪酸酯、聚甘油脂肪酸酯、脂肪酸双乙醇酰胺、聚氧乙烯山梨醇酐单硬脂酸酯、聚氧乙烯氢化蓖麻油和聚氧乙烯脂肪酸酯。这些发泡剂可以单独使用或它们中的两种或多于两种组合使用。
s.鞣剂
合适的鞣剂包括但不限于α-羟基醛和酮、甘油醛和相关的醇醛、各种吲哚、咪唑及其衍生物,以及各种认可的着色剂。其他合适的鞣剂包括但不限于丙酮醛、甘油醛、赤藓酮糖、四氧嘧啶、2,3-二羟基琥珀醛、2,3-二甲氧基琥珀醛、2-氨基-3-羟基-琥珀醛和2-苄氨基-3-羟基琥珀醛。
t.收敛剂
合适的收敛剂包括但不限于柠檬酸铝、乳酸铝、白桦提取物、咖啡提取物、月见草提取物、葡萄提取物、散沫花提取物、常春藤提取物、柠檬提取物、金缕梅提取物、铵和钾明矾、三聚磷酸铝、甘氨酸铝和酚硫酸铝、尿囊素氯羟铝、尿囊素羟铝、硬脂酸铝、硫酸铝和柠檬酸铝、磷酸钠铝、钠明矾、乳酸氯羟化铝钠、乳酸钙、氯化钙、硫酸钙水合物、乳酸铝钠、醋酸锌、氯化锌、硫酸锌、乳酸锌、锌沸石、苯酚磺酸锌及其组合。提取物是指整个水果、豆类和/或植物,或这些水果、豆类和/或植物的选择成分。
u.杀菌剂
合适的杀菌剂包括但不限于对羟苯甲酸的甲酯、对羟苯甲酸的乙酯、对羟苯甲酸的丙酯、或对羟苯甲酸的丁酯、苯氧乙醇、邻苯基苯酚、脱氢乙酸或其盐、对甲酚、间甲酚、邻氯间二甲苯酚、薄荷油、紫锥花、美洲血根草、辣椒、茶树精油、野生佛手柑(bergamont)、矮木丛、刺荨麻(stinging metal)、月桂、没药、拉檀根树皮、美洲花椒、金盏花、甘菊、莫匹罗星、硫酸新霉素、杆菌肽、多黏菌素B、左旋氧氟沙星、四环素(盐酸金霉素、盐酸土霉素和盐酸四环素)、克林霉素磷酸酯、硫酸庆大霉素、苯扎氯铵、苄索氯铵、己基间苯二酚、甲苄索氯铵、苯酚、季铵化合物、三氯卡班(triclocarbon)、三氯生和茶树油。
v.除臭剂和止汗剂
合适的除臭剂和止汗剂包括但不限于锌盐,例如硫酸锌和氯化锌、甘氨酸盐如甘氨酸铝锆、氯化氢铝、四氯甘氨酸铝锆、碳酸锌、邻苯基苯酚和季铵化合物如二甲基苄基氯化铵和氯化六甲双铵。
w.增白剂
皮肤增白剂的实例包括但不限于对苯二酚、曲酸、甘草和/或其衍生物、抗坏血酸和/或其衍生物、熊果苷、熊果提取物、光果甘草及其衍生物、小科藻科提取物、紫苏提取物、椰子果肉提取物和/或其他脱色剂。
x.抗微生物剂
抗微生物剂的实例包括但不限于三氯生,3,4,4′-三氯碳酰苯胺(三氯卡班);3,4,4′-三氟甲基-4,4′-二氯碳酰苯胺(卤卡班);5-氯-2-甲基-4-异噻唑啉-3-酮;碘丙炔丁基氨基甲酸酯;8-羟基喹啉;柠檬酸8-羟基喹啉;硫酸8-羟基喹啉;4-氯-3,5-二甲苯酚(氯二甲苯酚);2-溴-2-硝基丙烷-1,3-二醇;重氮烷基脲;布康唑;制真菌素;特康唑;呋喃妥因;非那吡啶;阿昔洛韦;克霉唑;氯二甲苯酚;洗必太;咪康唑;特康唑;对羟基苯甲酸丁酯;尼泊金丁酯;羟苯乙酯;对羟基苯甲酸甲酯;甲基氯异塞唑啉酮;甲基异噻唑啉;1,3-二(羟甲基)-5,5-二甲基海因和3-碘-2-丙炔基丁基氨基甲酸酯的混合物;羟基喹啉;EDTA;EDTA四钠;对羟基苯甲酸酯;烷基吡啶化合物;可可磷脂酰PG-氯化二铵;葡萄糖酸氯己定;葡萄糖酸氯己定;醋酸氯己定;羟乙基磺酸氯己定;盐酸氯己定;苯扎氯铵;苄索氯铵;聚六亚甲基双胍;及其混合物。
2.药物成分
还预期药物活性剂对本发明的组合物是有用的。药物活性剂的非限制性实例包括抗粉刺剂、用于处理酒渣鼻的试剂、止痛剂、麻醉剂、肛门直肠剂、抗组胺药、包括非甾族消炎药的消炎剂、抗生素、抗真菌剂、抗病毒素、抗微生物剂、抗癌活性剂、抗疥螨剂、灭虱剂、抗肿瘤药、止汗剂、止痒剂、抗牛皮癣药、抗脂溢剂、生物活性蛋白质和肽、烧伤处理剂、烧灼剂、脱色剂、脱毛剂、尿布疹处理剂、酶、毛发生长刺激剂、包括DFMO及其盐和类似物的毛发生长抑制剂、止血剂、角质分离剂、口疮处理剂、唇疱疹处理剂、牙科或牙周处理剂、光敏感活性剂、皮肤保护剂/屏障剂、包括激素和皮质激素的类固醇、晒伤处理剂、防晒剂、经皮活性剂、疣处理剂、创伤处理剂、创伤愈合剂等。
G.试剂盒
还预期用于本发明的某些方面的试剂盒。例如,本发明的组合物可以被包括在试剂盒内。试剂盒可以包括容器。容器可以包括安瓿、瓶子、金属管、层压管、塑料管、分配器、加压容器、屏障容器、袋、分室、口红容器、压缩容器、能够保存化妆品组合物的化妆品盘或其它类型的容器如注射或吹塑成型的塑料容器,其中保存分散体或组合物或期望的瓶子、分配器或包装。试剂盒和/或容器在其表面可包含标记。举例来说,标记可以是字词、短语、缩写、图片或符号。
所述容器可以分配预定量的组合物。在其他实施方案中,可以挤压容器(例如金属、层压或塑料管)以分配期望量的组合物。组合物可以分配为喷雾、气溶胶、液体、流体或半固体。容器可以具有喷雾、抽吸机构或挤压机构。试剂盒还可以包含使用试剂盒组分以及使用任何包含于容器内的组合物的说明书。说明书可以包含如何施用、使用和保存组合物的解释。
实施例
列出了以下实施例以说明本发明的优选实施方案。本领域技术人员应理解,以下实施例中所公开的技术代表本发明人发现的在本发明的实践中发挥良好作用的技术,并因此可以被认为为其实践建立优选的模式。然而,根据本公开,本领域技术人员应理解,在不脱离本发明的精神和范围的情况下,在所公开的具体实施方案中可以做许多改变,并仍获得相同或相似的结果。
本文所公开和要求保护的所有组合物和方法根据本公开可以不需要过多实验即而做出和实现。尽管本发明的组合物和方法已经按照优选实施方案进行了描述,但是对于本领域技术人员明显的是,在不脱离本发明的构思、精神和范围的情况下,可以对所述组合物和方法以及在本文所描述方法的步骤或步骤的顺序中进行改变。更具体地,明显地,化学和生理两方面都相关的特定试剂可以替代本文所描述的试剂,同时会实现相同或相似的结果。所有对本领域技术人员明显的这类相似的替代和改变都被视为在如由所附权利要求限定的本发明的精神、范围和构思内。
胶原蛋白是缔结组织的主要成分之一,并且有助于保持皮肤看起来年轻。胶原蛋白是人身体中最丰富的蛋白质,约占其蛋白质组成的三分之一。当胶原蛋白水平变高,皮肤更紧实、更光滑和更柔软。人的胶原蛋白产生在二十岁后开始下降,在二十岁时胶原蛋白生产以约每年1%的速率下降,皮肤逐渐变得更薄、更脆弱。随着年龄增加,嘴巴和眼睛周围开始形成细纹,前额的皱纹开始出现。
实施例1
(实施例制剂)
表1表示本发明的护肤液制剂,其被构建为水包油乳液。
表1*
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*制剂可以通过在室温或例如70℃至75℃加热下在烧杯中混合所述成分直到均匀来制备。然后,可以将制剂冷却到标准室温(20℃至25℃)。此外,如果需要,可以添加额外的成分,例如以改变为皮肤提供益处的组合物或成分的流变性质。可以添加赋形剂,例如以改变组合物的流变性质。或者,可以改变水的量,只要组合物中水的量为至少5重量/重量%,优选10重量/重量%至40重量/重量%。
实施例2
(体外肌管收缩数据)
发现迷迭香叶提取物、法国薰衣草提取物和千日菊提取物中的每一种都减少了肌管的收缩,这可以导致面部肌肉的肌肉收缩减少,和由于重复性面部肌肉收缩引起的面部深层细纹或皱纹以及动态面部皱纹和前额皱纹减少。用于获得数据的方法包括:(1)将原代人骨骼肌成肌细胞(HSMM)分化成肌管;(2)然后,用“诱发剂”(乙酰胆碱)处理肌管,继而引起钙流入并产生动作电位;和(3)该动作电位引起肌肉细胞收缩。表2提供了所使用的每种提取物的浓度量和得到的钙水平的抑制效果。
表2
*迷迭香叶提取物是共晶提取物,其中用于制备提取物的溶剂是甜菜碱、乳酸和水的流体混合物。所述提取物可以以商品名Rosemary EUTECTYS BLATM由Naturex(法国)提供。
**使用的法国薰衣草提取物是法国薰衣草花/叶/茎的超临界CO2提取物。该CO2提取物与辛酸/癸酸甘油三酯混合。该提取物以商品名Stochey’s由Barnet Products LLC(恩格尔伍德克利夫斯,新泽西(美国))提供。
***使用的千日菊提取物是水-乙醇提取物,其中溶剂包括水、乙醇和1,3丙二醇的流体混合物。从所得提取液体中除去乙醇。该提取物以商品名ExpressionAF由Gattefossé(法国)提供。
实施例3
(体外肌管收缩数据)
发现千日菊提取物提高了人真皮成纤维细胞中胶原蛋白的产生。表3提供了使用的提取物的数据和量。
表3
成分 | 胶原蛋白产量增加 |
千日菊提取物* | 1.0重量%浓度=胶原蛋白产量增加61% |
*使用的千日菊提取物是水-乙醇提取物,其中溶剂包括水、乙醇和1,3丙二醇的流体混合物。从所得提取液体中除去乙醇。该提取物以商品名ExpressionAF由Gattefossé(法国)提供。
胶原蛋白是对皮肤结构关键的一种细胞外基质蛋白。增加的胶原蛋白合成帮助改善皮肤坚实度和弹性。使用生物分析检验千日菊提取物在人真皮成纤维细胞产生前胶原肽(胶原蛋白前体)上的效果。该分析的终点是反映前胶原肽的存在和细胞活性的分光光度测量。该分析采用定量夹心酶免疫分析技术,因此对前胶原肽特异的单克隆抗体预先涂覆在微孔板上。将标准品和样品移液至孔中,存在的任何前胶原肽被固定化的抗体结合。洗去所有未结合的物质后,向孔中添加对前胶原肽特异的酶联多克隆抗体。冲洗以除去任何未结合的抗体-酶试剂之后,向孔中加入底物溶液,使颜色显现与最初步骤中结合的前胶原肽的量成比例。终止颜色显现,使用酶标仪测量在450nm处颜色的强度。
为了产生样品和对照,在37℃下10%的CO2中,用10%的胎牛血清(Mediatech)在标准DMEM生长培养基中培育亚汇合的正常成人真皮成纤维细胞(Cascade Biologics)。用每种测试成分和对照处理细胞3天。孵育之后,收集细胞培养基,并如上所述,使用来自Takara(#MK101)的夹心酶联免疫吸附实验(ELISA)定量前胶原肽的分泌量。
实施例4
(体外层黏连蛋白表达数据)
发现千日菊提取物提高了人真皮成纤维细胞中层黏连蛋白的产生。表4提供了使用的提取物的数据和量。
表4
成分 | 层黏连蛋白产量增加 |
千日菊提取物* | 1.0重量%浓度=层黏连蛋白产量增加53% |
*使用的千日菊提取物是水-乙醇提取物,其中溶剂包括水、乙醇和1,3丙二醇的流体混合物。从所得提取液体中除去乙醇。该提取物以商品名ExpressionAF由Gattefossé(法国)提供。
层黏连蛋白是真皮-表皮连接(DEJ)(也称为基膜)中的主要蛋白质。DEJ位于真皮和表皮之间,并连结形成叫做网脊的指状突起。表皮细胞从真皮的血管中吸收其养分。网脊增加了接触到这些血管和所需营养的表皮的表面积。DEJ提供两个组织隔室的黏连,并控制皮肤的结构完整性。层黏连蛋白是位于DEJ中的结构糖蛋白。考虑到将细胞保持在一起的胶,由真皮成纤维细胞分泌层黏连蛋白以帮助促进表皮细胞与DEJ的细胞内和细胞间黏附。通过量化在培育的人成纤维细胞的细胞上清液中的层黏连蛋白来监测层黏连蛋白的分泌,所述人成纤维细胞用具有测试成分(千日菊提取物)的培养基处理了3天。培育之后,在酶联免疫吸附测试(ELISA)中,使用针对每种蛋白质的免疫荧光抗体测量层黏连蛋白。对细胞代谢活性的测量进行标准化,其由3-(4,5-二甲基噻唑-2-基)-5-(3-羧基甲氧基苯基)-2-(4-磺苯基)-2H-四唑(MTS)的生物转化来测定。
实施例5
(体外MMP抑制数据)
发现迷迭香叶提取物抑制了人真皮成纤维细胞中的基质金属蛋白酶(MMP)1、基质金属蛋白酶3和基质金属蛋白酶9。表5提供了使用的提取物的数据和量。
表5
*迷迭香叶提取物是共晶提取物,其中用于制备提取物的溶剂是甜菜碱、乳酸和水的流体混合物。所述提取物可以以商品名Rosemary EUTECTYS BLATM由Naturex(法国)提供。
基质金属蛋白酶1(MMP-1)的酶活性分析:MMP是胞外蛋白酶,凭借其广泛的底物特异性在许多正常和疾病状态中起作用。MMP-1底物包括IV型胶原蛋白。分子探针Enz/Chek白明胶酶/胶原酶分析试剂盒(#E12055)利用荧光明胶底物来检测MMP1蛋白酶活性。在溶蛋白性裂解后,显示亮绿色荧光,并可以使用荧光酶标仪来观测亮绿色荧光以测量酶活性。将来自Invitrogen的Enz/Chek白明胶酶/胶原酶分析试剂盒(#E12055)设计为体外分析以测量MMP-1酶活性。分析迷迭香叶提取物。该分析依赖于纯化的MMP-1酶降解荧光明胶底物的能力。一旦底物被MMP-1特异裂解,则显示亮绿色荧光并使用荧光酶标仪来监测。在纯化的酶和底物存在或不存的条件下培育迷迭香叶提取物以确定其蛋白酶抑制能力。
基质金属蛋白酶3和基质金属蛋白酶9(MMP3和MMP9)的酶活性分析:MMP-3底物包括胶原蛋白、纤连蛋白和层黏连蛋白;MMP9底物包括VII型胶原蛋白、纤连蛋白和层黏连蛋白。使用来自BioMol International的用于MMP3(AK-400)和MMP-9(AK-410)的比色药物发现试剂盒,该分析设计用于其使用含硫多肽作为显色底物(Ac-PLG-[2-巯基-4-甲基-戊酰基]-LG-OC2H5)5,6来测量MMP的蛋白酶活性。MMP裂解位点的肽键由含硫多肽中的硫酯键替代。MMP对该键的水解产生了巯基,该巯基与DTNB[5,5'-二硫代双(2-硝基苯甲酸),埃尔曼试剂]反应形成2-硝基-5-硫代苯甲酸,这可通过在412nm(ε=13600M-1cm-1在pH 6.0及7以上)处的吸光度进行检测。分析迷迭香叶提取物。
实施例6
(体内临床数据)
获得了迷迭香叶提取物、法国薰衣草提取物和千日菊提取物的组合的体内临床数据。迷迭香叶提取物是共晶提取物,其中用于制备提取物的溶剂是甜菜碱、乳酸和水的流体混合物。所述提取物可以以商品名ROSEMARY EUTECTYS BLATM由Naturex(法国)提供。使用的法国薰衣草提取物是法国薰衣草花/叶/茎的超临界CO2提取物。该CO2提取物与辛酸/癸酸甘油三酯混合。该提取物以商品名STOCHEY’S由Barnet Products LLC(恩格尔伍德克利夫斯,新泽西(美国))提供。使用的千日菊提取物是水-乙醇提取物,其中溶剂包括水、乙醇和1,3丙二醇的流体混合物。从所得提取流体中除去乙醇。该提取物以商品名EXPRESSION AF由Gattefossé(法国)提供。
体内临床研究的目的是评估三种提取物的组合在30个人(n=30)中在四周内每日施用两次的情况下,对松弛肌肉和减少皱纹的功效。将三种提取物的组合置于皮肤病学可接受的载体上。皮肤病学可接受的载体设计用于测试三种提取物的组合的功效。相信皮肤病学可接受的载体中的其他成分不会对临床数据结果产生积极或消极影响。皮肤病学可接受的载体中使用的每种提取物的量为1重量%的迷迭香叶提取物、2重量%的法国薰衣草提取物和2重量%的千日菊提取物。
体内临床研究设计为随机对照、研究者盲法临床研究。使用以下参数:(1)对象——年龄为35岁至70岁的身体状况良好(无需体检)的女性并且经临床确定,在10分的范围内中度皱眉纹/眉间纹评分为3-7分(包含地);(2)测试位点——眉间区域;(3)评估时间点——基线(第0周)、第2周、第4周和第8周;(4)测试方法——用于肌肉响应(mV/s):AUC(曲线下的面积)的肌电描记术(EMG);和(5)测量的临床终点——0至9范围内的细纹和皱纹以及皮肤纹理。
图1至图2提供了通过EMG测试表示肌肉收缩的抑制的数据。图3至图4提供了表示细纹和皱纹的外观减轻的数据。如这些图1至图4中所示,数据表明迷迭香叶提取物、法国薰衣草提取物和千日菊提取物可以减少面部肌肉收缩并可以减轻细纹和皱纹的整体外观。以下表6提供了表示著名的可注射的神经调质((onabotulinumtoxinA))与迷迭香叶提取物、法国薰衣草提取物和千日菊提取物的组合之间的EMG响应和临床对比的数据。
表6
*表示p<0.05,当与基线相比,改善具有统计显著性。
**将BoTox-5U用作对照以验证方法并在5个人上进行。每个人对眉间区域施用5单位(U)的Botox并测量EMG响应(AUC)%。这样做是为了使上述提取物组合的研究标准化。
实施例7
(体内临床数据)
在对比临床研究中,获得了迷迭香叶提取物、法国薰衣草提取物和千日菊提取物的组合的体内临床数据。迷迭香叶提取物是共晶提取物,其中用于制备提取物的溶剂是甜菜碱、乳酸和水的流体混合物。所述提取物可以以商品名ROSEMARY EUTECTYS BLATM由Naturex(法国)提供。使用的法国薰衣草提取物是法国薰衣草花/叶/茎的超临界CO2提取物。该CO2提取物与辛酸/癸酸甘油三酯混合。该提取物以商品名STOCHEY’S由BarnetProducts LLC(恩格尔伍德克利夫斯,新泽西(美国))提供。使用的千日菊提取物是水-乙醇提取物,其中溶剂包括水、乙醇和1,3丙二醇的流体混合物。从所得提取流体中除去乙醇。该提取物以商品名EXPRESSION AF由Gattefossé(法国)提供。
对比体内临床研究的目的是评估三种提取物的组合对实验组1和实验组2参与者实施在四周内每天两次的肌肉收缩和皱纹的减少的功效,所述实验组1使用了微型滚轮(n=33),实验组2参与者不使用微型滚轮(n=30)。将三种提取物的组合置于皮肤病学可接受的载体上作为测试产品。皮肤病学可接受的载体设计用于测试三种提取物的组合在使用微型滚轮施用和不使用微型滚轮施用时的功效。实验组1的研究参与者使用了1.0mm的微型滚轮。研究参与者在用私人的洗面奶清洁他们的脸之后,早上和晚上在其眉毛之间施用含有三种提取物的组合物。对于实验组1,研究参与者在晚上每周使用3次微型滚轮,在眉毛之间来回滚动微型滚轮4次至5次,每次通过时改变方向。对于实验组1和实验组2,将封闭贴片(例如)置于眉间整夜。相信皮肤病学可接受的载体中的其他成分不会对临床数据结果产生积极或消极影响。皮肤病学可接受的载体中使用的每种提取物的量为1重量%的迷迭香叶提取物、2重量%的法国薰衣草提取物和2重量%的千日菊提取物。
对比体内临床研究设计为随机对照、研究者盲法临床研究。使用以下参数:(1)对象——年龄为35岁至70岁的身体状况良好(无需体检)的女性并且经临床确定,在10分的范围内中度皱眉纹/眉间纹评分为3-7分(包含地);(2)测试位点——眉间区域;(3)测试产品——前额;补充的产品——封闭贴片和微型滚轮;(4)评估时间点——基线(第0周)、第2周、第4周和第8周;(5)测试方法——用于肌肉响应(mV/s):AUC(曲线下的面积)的肌电描记术(EMG);和(6)测量的临床终点——0至9范围内的细纹和皱纹以及皮肤纹理。
图5至图6提供了通过EMG测试表示肌肉收缩抑制的数据,比较了使用微型滚轮和不使用微型滚轮的施用之间的改善百分比。图7至图8提供了表示细纹和皱纹的外观减轻的数据,比较了使用微型滚轮和不使用微型滚轮的施用之间的改善百分比。如图5至图8中所示,数据表明,含有迷迭香叶提取物、法国薰衣草提取物和千日菊提取物的组合的组合物的施用,其与使用微型滚轮结合以促进组合物向皮肤内渗透,这可以减少面部肌肉收缩并且可以减轻细纹和皱纹的整体外观。经确定,与基线相比,在第2、4周和8周使用微型滚轮的组合物的施用有效实现了更快和更强的面部肌肉收缩减少,并在4周内减少细纹和皱纹以及皮肤纹理,其中在8周后79%的对象在细纹和皱纹以及皮肤纹理上均显示出改善。还确定了在8周后不使用微型滚轮的组合物的施用仍有效实现了面部肌肉收缩的减少,并在4周内减少细纹和皱纹以及皮肤纹理,其中在8周后87%的对象在细纹和皱纹上均显示出改善,90%的对象在皮肤平滑度上显示出改善。
图9至图10提供了表示改善皮肤纹理/粗糙度的数据,比较了使用微型滚轮和不使用微型滚轮的施用之间的改善百分比。如图9至图10中所示,数据表明,含有迷迭香叶提取物、法国薰衣草提取物和千日菊提取物的组合物的施用,其与使用微型滚轮结合以促进组合物向皮肤内渗透,这可以在第2周、4周和8周后在眉间区域显著改善皮肤纹理和粗糙度。经确定,79%的对象在用微型滚轮施用组合物的8周后显示出皮肤平滑度有所改善。还确定了不使用微型滚轮施用组合物仍可在8周后有效改善皮肤平滑度。
以下表7提供了表示著名的可注射的神经调质((onabotulinumtoxinA))与用微型滚轮施用和不用微型滚轮施用的迷迭香叶提取物、法国薰衣草提取物和千日菊提取物的组合之间的EMG响应和临床对比的数据。
表7
*表示p<0.05,当与基线相比,改善具有统计学显著性。
**在比较实验组1和实验组2时,将BoTox-5U用作阳性对照并在5个人上进行。每个人对眉间区域施用5单位(U)的Botox并测量EMG响应(AUC)%。这样做是为了使上述提取物组合的研究标准化。
实施例8
(附加分析)
可用于确定在本说明书全文和权利要求书中所公开的任意成分或任意的成分组合或具有所述成分组合的组合物功效的附加分析,可以通过本领域普通技术人员已知的方法进行确定。以下是可以在本发明的环境中使用的非限制性分析。应认识到,可以使用其它测试过程,包括例如客观的和主观的过程。
弹性蛋白刺激分析:弹性蛋白是结缔组织蛋白,其帮助皮肤在伸展或收缩后恢复形状。弹性蛋白也是重要的负载蛋白,其用于需要储存机械能之处。弹性蛋白通过在由赖氨酰氧化酶催化的反应中连接许多可溶的弹性蛋白原蛋白分子来产生。可以通过直接ELISA夹心法在培养的人成纤维细胞中监测弹性蛋白分泌和弹性蛋白纤维,并使用Meso ScaleDiscovery系统SECTOR 2400成像系统进行分析。
层黏连蛋白和纤连蛋白刺激分析:层黏连蛋白和纤连蛋白是真皮-表皮连接(DEJ)(也称为基膜)中主要的蛋白质。DEJ位于真皮和表皮连结之间,连结形成指状突起,称为网脊。表皮细胞从真皮的血管中接收其养分。网脊增大了暴露于这些血管和所需养分的表皮的表面积。DEJ提供两个组织隔室的黏连,并控制皮肤的结构完整性。层黏连蛋白和纤连蛋白是位于DEJ中的两种结构糖蛋白。考虑到将细胞保持在一起的胶,真皮成纤维细胞分泌层黏连蛋白和纤连蛋白以帮助促进表皮细胞到DEJ的细胞内和细胞间黏连。
层黏连蛋白和纤连蛋白的分泌可以通过定量在培养的人成纤维细胞的细胞上清液中的层黏连蛋白和纤连蛋白来监测,培养的人成纤维细胞用含或不含1.0%最终浓度的测试成分的培养基处理3天。培养之后,在酶联免疫吸附测试(ELISA)中,可以使用针对每种蛋白质的免疫荧光抗体来测定层黏连蛋白和纤连蛋白含量。
ORAC分析:还可以通过测量这种成分或组合物的抗氧化活性来分析本说明书公开的活性成分、成分的组合或具有所述组合的组合物中任意一种的氧自由基吸收(或吸收率)能力(ORAC)。抗氧化活性说明减少氧化试剂(氧化剂)的能力。该分析定量抑制氧化剂,例如已知对细胞(例如皮肤细胞)造成损害的氧自由基的作用的程度及所需时长。本说明书公开的活性成分、活性成分的组合或含有所述组合的组合物中的任一种的ORAC值可以通过本领域普通技术人员已知的方法进行确定(参见U.S.公开号2004/0109905和2005/0163880;和商业上可获得的试剂盒如Zen-Bio ORAC抗氧化分析试剂盒(#AOX-2))。Zen-Bio ORAC抗氧化分析试剂盒测量了由于通过AAPH(2,2'-偶氮二-2-甲基丙基咪二盐酸盐)分解而形成过氧化氢-自由基的随时间的荧光素荧光的损失。Trolox(水溶性维生素E类似物)以剂量依赖的方式作为抑制荧光素衰变的阳性对照。
基质金属蛋白1酶活性(MMP1)测定:体外基质金属蛋白酶(MMP)抑制分析。MMP是胞外蛋白酶,其凭借广泛的底物特异性对许多正常的和疾病状态起作用。MMP1底物包括胶原蛋白IV。分子探针Enz/Chek白明胶酶/胶原酶分析试剂盒(#E12055)利用荧光明胶底物来检测MMP1蛋白酶活性。在溶蛋白性裂解后,显示亮绿色荧光,并可以使用荧光酶标仪来观测亮绿色荧光以测量酶活性。
将来自Invitrogen的Enz/Chek白明胶酶/胶原酶分析试剂盒(#E12055)设计为体外分析以测量MMP1酶活性。可以分析说明书中公开的活性成分、任意一种成分的组合、或具有所述组合的组合物。该分析依赖于纯化的MMP1酶降解荧光明胶底物的能力。一旦底物被MMP1特异地裂解,则显示亮绿色荧光,并可以使用荧光酶标仪观测亮绿色荧光。在纯化的酶和底物存在或不存的条件下孵育测试物质以确定其蛋白酶抑制能力。
环氧合酶(COX)测定:体外环氧合酶-1和-2(COX-1、-2)抑制分析。COX是展现环氧合酶和过氧化物酶两者活性的双功能酶。环氧合酶活性将花生四烯酸转换为过氧化氢内过氧化物(前列腺素G2;PGG2),过氧化物酶组分将内过氧化物(前列腺素H2;PGH2)还原为相应的醇,前列腺素、凝血烷和环前列腺素的前体。该COX抑制剂筛选分析测量环氧合酶的过氧化物酶组分。过氧化物酶活性通过监测氧化的N,N,N',N'-四甲基-对苯二胺(TMPD)的出现进行比色分析。该抑制筛选分析包含COX-1和COX-2酶两者,以筛选同工酶特异性抑制剂。可以使用比色COX(绵羊)抑制剂筛选分析(#760111,Cayman Chemical)来分析本说明书中所公开的每一种活性成分、任一种成分的组合、或具有所述组合的组合物对纯化的环氧合酶(COX-1或COX-2)活性的影响。根据制造商的说明,纯化酶、血红素和测试提取物可以在分析缓冲液中混合,并在室温下摇晃孵育15分钟。培育之后,可以加入花生四烯酸和比色底物开始反应。颜色变化可以通过在590nm读取的比色板进行评估。COX-1或COX-2活性的抑制百分比可以通过与未处理的对照物比较进行计算,以确定试验提取物抑制经纯化的酶活性的能力。
脂肪氧合酶(LO)分析:体外的脂肪氧合酶(LO)抑制分析。LO是非血红素的含铁的加双氧酶,其催化分子氧加成到脂肪酸。亚油酸盐/酯和花生四烯酸盐/酯是植物中和动物中LO的主要底物。然后可以将花生四烯酸转化为羟基二十碳四烯(HETE)酸衍生物,随后将其转化为有效的炎症介质白三烯。该分析通过测量利用花生四烯酸培养的脂肪氧合酶(5-、12-或15-LO)产生的氢过氧化物提供精确且方便的用于筛选脂肪氧合酶抑制剂的方法。比色的LO抑制剂筛选试剂盒(#760700,Cayman Chemical)可以用于确定说明书中所公开的活性成分、任意一种成分的组合、或具有所述组合的组合物的每一种抑制酶活性的能力。可以将经纯化的15-脂肪氧合酶和测试成分在分析缓冲液中混合,并在室温下摇动培育10分钟。培育之后,可以加入花生四烯酸开始反应,并且混合物可以在室温下再培育额外的10分钟。可以加入比色底物终止催化,颜色变化可以通过在490nm测定的荧光板进行评估。脂肪氧合酶活性的抑制百分比可以通过与未处理的对照物对比来计算,以确定说明书中所公开的每种活性成分、任意一种成分的组合、或具有所述组合的组合物中的每一种抑制经纯化的酶活性的能力。
弹性蛋白酶分析:来自Molecular Probes(俄勒冈州尤金,美国)的弹性蛋白酶分析(试剂盒#E-12056)可用作体外酶抑制分析以测量本说明书中所公开的活性成分、任一种成分的组合或具有所述组合的组合物中每一种的弹性蛋白酶活性的抑制。EnzChek试剂盒包含可用染料标记的可溶性牛颈韧带弹性蛋白,从而使缀合物的荧光猝灭。非荧光的底物可以被弹性蛋白酶或其他蛋白酶消化以产生高荧光的片段。产生的荧光增强可以用荧光微孔板测定仪进行监测。来自弹性蛋白底物的消化产物在约505nm处具有最大吸收,在约515nm处具有最大荧光发射。当使用EnzChek弹性蛋白酶分析试剂盒以筛选弹性蛋白酶抑制剂时,可以将肽、N-甲氧基琥珀酰-Ala-Ala-Pro-Val-氯甲基酮用作选择性的、共同的弹性蛋白酶抑制剂。
控油分析:用于测量皮脂腺中皮脂分泌的减少和/或皮脂腺中皮脂产生的减少的分析可以通过使用本领域普通技术人员已知的标准技术进行分析。在具体的实例中,可以使用前额。可以将本说明书中所公开的活性成分、任意一种成分的组合或具有所述组合的组合物中的每一种,每天一次或两次地施用至前额的一部分一段固定的日子(例如1、2、3、4、5、6、7、8、9、10、11、12、13、14或多于14天),而前额的其他部分不用所述组合物处理。在一段固定的天数期满后,皮脂分泌可以通过将优质吸墨纸施用至处理过的和未处理的前额皮肤上进行分析。这是通过首先用湿的和干的布从处理过的和未处理的区域除去所有皮脂完成的。然后可以将吸油纸施用至处理过的和未处理的前额区域,并且可以围绕前额放置橡皮筋以轻轻地将吸油纸压到皮肤上。2小时后,吸墨纸可以移去,使其干燥,然后进行透照。越深的吸墨纸对应于越多的皮脂分泌(或较浅的吸墨纸对应于减少的皮脂分泌)。
红斑分析:测量皮肤发红减少的分析可以使用Minolta色度计进行评估。皮肤红斑可以通过在对象前臂施用0.2%的十二烷基硫酸钠溶液来引发。该区域用封闭的贴片保护24小时。24小时后,除去贴片,可以使用Minolta Chroma Meter的a*值对刺激诱发的发红进行评估。a*值测量肤色在红色区域的变化。读数后立即用本说明书公开的活性成分、活性成分组合中的任一种或含有所述组合的组合物处理区域。可定期进行重复测量以确定制剂减少发红和刺激的能力。
皮肤水分/水合分析:皮肤水分/水合的有益效果可以通过利用以Nova DermalPhase Meter进行的阻抗测量进行测量。阻抗计测量皮肤水分含量的变化。皮肤外层具有不同的电性质。当皮肤干燥时,其导电很差。当其变得更加含水时,产生增加的导电性。因此,皮肤阻抗(与导电性有关)的变化可以用于评价皮肤水合的变化。装置可以根据仪器说明针对每个测试日进行校准。还可以对温度和相对湿度进行标记。对对象可以进行如下评估:测量前其可以在具有确定湿度(例如30%至50%)和温度(例如68℃至72℃)的室内进行平衡。在脸的每一侧进行三个独立的阻抗测定,并对其进行记录和平均。阻抗计可以使用T5设定,其对施用至脸上每五秒的阻抗值进行平均。变化可以以统计方差和显著性进行报道。根据这个过程可以分析本说明书中公开的活性成分、任意一个成分组合中或具有所述组合的组合物中的每一种。
皮肤清透度和雀斑与老年斑减少的分析:可以使用Minolta色度计评估皮肤清透度和雀斑与老年斑的减少。肤色的变化可以使用Minolta Chroma Meter的a*值进行评估以确定由于产品处理引起刺激的可能性。a*值测量肤色在红色区域的变化。这用于确定本说明书中公开的活性成分、任意一种成分的组合、或具有所述组合的组合物中的每一种是否诱导刺激。测量可以在脸的每一侧进行并进行平均,作为左边和右边脸的值。皮肤清透度也可以使用Minolta Meter进行测量。测量是Minolta Meter的a*、b和L值的组合,并与皮肤的亮度有关,而且很好地对应皮肤的光滑度和水合。皮肤测定如上进行。在一个非限制性方面,皮肤清透度可以描述为L/C,其中C是色度并定义为(a2+b2)1/2。
皮肤干燥、表面细纹、皮肤光滑度和肤色分析:皮肤干燥、表面细纹、皮肤光滑度和肤色可以用临床评分技术进行评估。例如,皮肤干燥的临床评分可以通过五点标准Kligmanscale进行确定:(0)皮肤是柔软和湿润的;(1)皮肤呈现正常而没有可见的干燥;(2)皮肤触摸感觉轻微干燥而没有可见的剥落;(3)皮肤感觉干燥、坚硬并且具有带有一些鳞屑的发白的外观;以及(4)皮肤感觉非常干燥、粗糙并且具有带有鳞屑的发白的外观。评估可以由两个临床医师独立进行并取平均分。
肤色临床评分分析:肤色的临床评分可以通过十点模拟数值量表实施:(10)平滑均匀的皮肤、粉棕色的颜色。手持放大镜检查时没有暗的、红色或有鳞状的斑块。皮肤的细微纹理摸上去非常均匀;(7)不用放大镜观察均匀的肤色。没有鳞状区域,但是有由于色素沉着或红斑引起的疹斑。没有直径大于1cm的疹斑;(4)轻易地注意到皮肤疹斑和不均匀的纹理。少量脱皮。皮肤的某些区域摸上去粗糙;以及(1)不均匀的皮肤着色和纹理。多个区域的脱皮和变色,色素减退型、红色的或黑色的斑点。大面积的直径超过1cm的不均匀着色。评估可以由两个临床医师独立进行并取平均分。
皮肤平滑度的临床评分分析:皮肤光滑度的临床评分可以通过十点模拟数值量表进行分析:(10)光滑的,皮肤是湿润的和有光泽的,手指划过表面时没有阻力;(7)一定程度光滑的,微小的阻力;(4)粗糙的、可见地改变的,摩擦时有摩擦力;以及(1)粗糙的、起皮屑的、不均匀的表面。评估可以由两个临床医师独立进行并取平均分。
用Packman等人(1978)公开的方法进行的皮肤平滑度和皱纹减少的分析:皮肤光滑度和皱纹的减少也可以通过使用Packman等人(1978)公开的方法进行可视化评估。例如,每个对象就诊时,对每个对象的外表面线(SFL)的深度、浅度和总数量都可以进行认真的评分和记录。通过将数量因子乘以深度/宽度/长度因子得到数字的分数。获得眼睛区域和嘴巴区域(左侧和右侧)的分数,加到一起作为总的皱纹分数。
用Hargens Ballistometer进行的皮肤紧致度分析:皮肤紧致度可以用HargensBallistometer来测量,其是通过使小物体落在皮肤上并记录前两个反弹峰来评估皮肤弹性和紧致度的装置。Ballistometry是使用相对钝的探头(4平方毫米-接触面积)的小的轻量探针。探针轻轻地穿透进入皮肤,获得取决于皮肤外层性质的测量值,所述皮肤外层包括角质层和外表皮以及部分真皮层。
用气体轴承电测力计进行的皮肤柔软度/柔韧性分析:皮肤柔软度/柔韧性可以使用气体轴承电测力计(Gas Bearing Electrodynamometer)进行评估,其为测量皮肤应力/应变性质的仪器。皮肤的黏弹性与皮肤保湿有关。可以通过用双面胶将探针黏附在皮肤表面实现对脸颊区域预设位点的测量。大约3.5gm的力平行施加于皮肤表面,精确地测量皮肤的位移。然后可以计算皮肤的柔韧性,并以DSR(动态弹簧刚度,以gm/mm计)表示。
用复制品进行的纹路和皱纹的显现分析:皮肤上纹路和皱纹的显现可以使用复制品进行评估,所述复制品是皮肤表面的印模。可以使用如硅橡胶的材料。复制品可以通过图像分析进行分析。纹路和皱纹可见性的变化可以通过利用硅复制品形成对象的脸并用计算机图像分析系统分析复制品图像进行客观地定量。复制品可以从眼睛区域和颈部区域获得,并用数码相机以低角度入射照明进行拍摄。数字图像可以用图像处理程序进行分析。
用轮廓仪/记录针(Stylus)进行的皮肤表面轮廓分析:皮肤表面轮廓可以通过使用轮廓仪/记录针的方法进行测量。这包括闪光或拖动记录针穿过复制品表面。记录针的垂直位移通过距离传感器可以录入计算机,在扫描复制品的一定距离后,皮肤轮廓的曲线可以以二维曲面形式产生。该扫描可以沿着固定的轴重复任意次数以产生模拟的皮肤3-D图像。使用记录针技术可以获得十个随机的复制品截面,并组合产生平均值。感兴趣的值包括Ra,其为通过积分相对于平均轮廓高度的轮廓高度计算得到的所有粗糙度(高度)值的算术平均数。Rt,其为最高峰和最低谷之间的最大垂直距离,以及Rz,其为平均峰振幅减去平均峰高度。数值以用mm为单位标定的数值给出。设备应在每次使用前通过扫描已知数值的金属标准品进行归一化。Ra值可以通过下式计算:Ra=归一化粗糙度;lm=横向(扫描)长度;以及y=曲线位置相对于平均曲线高度(x轴)的绝对值。
MELANODERMTM分析:在其它非限制性方面,可以通过使用皮肤类似物例如MELANODERMTM来评价说明书中公开的活性成分、任意一种成分的组合或具有所述组合的组合物中每一种的功效。黑素细胞,皮肤类似物中细胞的一种,当暴露于黑色素前体L-二羟苯基丙氨酸(L-DOPA)时被明显地染色。皮肤类似物MELANODERMTM可以如下处理:用包含说明书中公开的活性成分、任意一种成分的组合或具有所述组合的组合物中的每一种的多种基质来处理,或仅用基质来处理作为对照。或者,未处理的皮肤类似物样品可以用作对照物。
丝聚合蛋白的产生:可测定由于本说明书中所公开的活性成分、任意一种成分组合或具有所述组合的组合物中每一种引起的角化细胞中丝聚合蛋白的产生的变化。丝聚合蛋白是皮肤中天然保湿因子(NMF)的前体。增加的NMF提高了皮肤中的水分含量。使用分析角化细胞溶解产物中的丝聚合蛋白浓度的生物测试来确定在经处理和未经处理的角化细胞中的丝聚合蛋白的产生。可用于量化丝聚合蛋白产生的生物分析的非限制性实例为SimonTM免疫印迹方案。对于每个样品,使正常的人表皮角化细胞(NHEK)在来自Life Technologies的含有钙的EPI-200-Mattek/>生长培养基(M-EP-500-CA)中生长。在处理之前,在37℃下5%的CO2中,使NHEK在生长培养基中孵育过夜。然后将NHEK在含有1%测试化合物/提取物的生长培养基中或不含化合物/提取物(阴性对照)的生长培养基中培育24小时至36小时。然后可将NHEK清洗、收集并储存于冰或较冷的环境中直至用裂解缓冲液和超声在冰上裂解。可以测定样品的蛋白质浓度并用于使样品归一化。裂解物可在-80℃下存储直至在定量分析中使用。
SimonTM蛋白质免疫印迹法使用定量的免疫印迹免疫分析技术,该技术使用了对丝聚合蛋白特异的抗体来定量检测样品中的丝聚合蛋白。将细胞样品裂解并对蛋白质浓度进行归一化。然后可将标准化的试样和分子量标准样使用毛细管电泳装载于变性蛋白质分离凝胶并在其上运行。使用针对丝聚合蛋白特异性的初代抗体对凝胶内的蛋白质进行固定以及免疫探针检测。然后用结合至初代抗体的酶联检测抗体对经固定的蛋白质进行免疫探针检测。然后可向经固定的蛋白质中添加化学发光底物溶液以使得化学发光显色与在固定中结合的丝聚合蛋白的量成比例。在特定的时间终止化学发光显色,可测定化学发光信号的强度并与阳性对照和阴性对照比对。
闭合蛋白的产生:可测定由于本说明书中所公开的活性成分、任意一种成分的组合的或具有所述组合的组合物中的每一种而引起的角化细胞中闭合蛋白产生的变化。闭合蛋白是对于紧密连结的形成和皮肤水分屏障功能重要的蛋白质。在处理的和未处理的角化细胞中如何确定闭合蛋白产生的非限制性实例为通过使用分析在角化细胞裂解物中的闭合蛋白浓度的生物分析。通过使用SimonTM蛋白质免疫印迹法进行该生物分析。对于样品,在37C下和5%的CO2中,来自Life Technologies的人表皮角化细胞(HEKa)(C-005-5C)在Epilife生长培养基中生长24小时,培养基中含有来自Life Technologies的钙(M-EP-500-CA),并补充有来自Life Technologies的Keratinocyte Growth Supplement(HKGS)(S-101-5)。然后使用测试化合物/提取物、作为阴性对照的无化合物/提取物或作为阳性对照的1mM的CaCl2,使HEKa在生长培养基中孵育24小时至48小时。然后,洗涤HEKa细胞,收集并储存在冰上或较冷的地方,直到使用裂解缓冲液在冰上裂解并进行超声处理。可确定试样的蛋白质浓度并用于标准化试样。裂解物在-80℃下存储直至在生物测试中使用。
SimonTM蛋白质免疫印迹法生物分析使用定量的蛋白质免疫印迹分析技术,该技术使用了对闭合蛋白特异的抗体来定量检测样品中的闭合蛋白。将细胞样品裂解并对蛋白质浓度进行归一化。然后将归一化的样品和分子量标准品用毛细管电泳负载在变性的蛋白质分离凝胶上运行。然后,使用针对闭合蛋白特异性的初代抗体对凝胶内的蛋白质进行固定以及免疫探针检测。用结合至初代抗体的酶联检测抗体对经固定的蛋白质进行免疫探针检测。然后向经固定的蛋白质中添加化学发光底物溶液以使得化学发光显色与在固定中结合的闭合蛋白的量成比例。可在特定的时间终止化学发光显色,可测定化学发光信号的强度并与阳性对照和阴性对照比对。
角化细胞单层渗透性:可测定由于本说明书中所公开的活性成分、任意一种成分的组合或具有该组合的组合物中的每一种而导致的角化细胞单层渗透性变化。角化细胞单层的渗透性是皮肤屏障的完整性的量度。作为非限制性实例,可以使用Millipore(ECM642)的体外血管渗透性分析测定在经处理和未经处理的角化细胞中的角化细胞单层渗透性。该分析分析了内皮细胞吸附、输送和渗透。简略来说,来自Life Technologies的成人表皮角化细胞(C-005-5C)可被接种到在收集孔内的多孔胶原蛋白涂覆的膜上。在37℃下和5%的CO2中,角化细胞在Epilife生长培养基中培养24小时,Epilife生长培养基中含有来自LifeTechnologies的钙(M-EP-500-CA),补充有来自Life Technologies的KeratinocyteGrowth Supplement(HKGS)(S-101-5)。培养时间使得细胞形成单层并封闭膜孔。然后将培养基替换成含测试化合物/提取物(测试试样)或不含测试化合物/提取物(未处理对照)的新鲜培养基,该角化细胞在37℃下和5%的CO2中培养另外48小时。在含/不含测试化合物/提取物的培养之后,为了测定角化细胞单层的渗透性,培养基被替换成含有高分子量异硫氰酸荧光素(FITC)-Dextran的新鲜的培养基,该角化细胞在37℃下和5%的CO2中培养4小时。在该4小时的培养期间,FITC可以与单层膜的渗透性成比例的速率穿过该角化细胞单层和多孔膜进入收集孔。在该4小时的培养之后,可测定细胞活性和收集孔中FITC的含量。对于FITC含量,收集孔中的培养基被收集,并且在520nm处激发时在480nm(Em)处测定培养基的荧光。可通过以下公式确定与未处理的对照组相比,渗透百分比和百分比的变化:渗透率百分比=((测试样品的Ex/Em平均值)/未经处理的对照的Ex/Em平均值)×100;变化百分比=测试样品的渗透率百分比-未经处理的对照的渗透率百分比。
透明质酸的产生:可测量在人真皮成纤维细胞中的由于本说明书中所公开的活性成分、任意一种的成分组合或具有所述组合的组合物中每一种而导致的透明质酸产生的变化。HA是参与基质结构的稳定性的多糖,其与向组织和细胞提供膨压有关。作为一个非限制性实例,可使用R&D Systems的Hyaluronan DuoSet ELISA试剂盒(DY3614)确定在经处理的和未经处理的成人真皮成纤维(HDFa)细胞中的HA的产生。在该分析中,对于试样的产生,在处理之前,将来自Cascade Biologics的近汇合HDFa细胞(C-13-5C)在37℃和10%的CO2下在饥饿培养基中(在Dulbecco氏改良Eagle培养基中含有0.15%的牛胎儿血清和1%的青霉素链霉素溶液)培养72小时。然后用含有测试化合物、阳性对照(来自Sigma-Aldrich的佛波醇12-十四酸酯13-乙酸酯(P1585)和来自Sigma-Aldrich的血小板衍生生长因子的(P3201))或无添加对照的新鲜饥饿培养基培养该细胞24小时。然后收集培养基并在-80℃下冷冻直至在ELISA测试使用。
简而言之,该ELISA分析采用定量的夹心酶免疫测定技术,因此对HA特异的捕获抗体可以预先涂覆在微孔板上。将标准品、来自经处理的细胞和未处理的细胞的培养基移液至微孔板,以使存在的任何HA被固定化抗体结合。冲走所有未结合的物质后,添加对HA特异的酶联检测抗体到孔中。冲洗以除去所有未结合的抗体-酶试剂之后,添加底物溶液到孔中,以使颜色显现与最初步骤中结合的HA的量成比例。在特定的时间终止颜色显现,可以使用酶标仪在450nm处测量颜色的强度。
透明质酸酶活性的抑制:可测定由于本说明书中所公开的活性成分、任意一种成分的组合或具有该组合的组合物的每一种而引起的透明质酸酶活性的变化。透明质酸酶是分解HA的酶。HA是参与基质结构的稳定性的多糖,其参与向组织和细胞提供膨压有关。作为一个非限制性实例,可以使用由Sigma-Aldrich方案#EC 3.2.1.35改良的体外方案测定透明质酸酶活性。简略来说,向含有测试化合物或对照组的微孔板反应孔中添加来自Sigma-Aldrich的1-S型透明质酸酶(H3506)。可使用鞣酸作为阳性对照抑制剂,对于对照的酶可以不添加测试化合物,可使用含有测试化合物的孔或并没有透明质酸酶的阳性对照作为背景的阴性对照。在底物(HA)的添加之前,孔在37℃下培养10分钟。添加底物,反应在37℃下培养45分钟。然后,将每个反应溶液的一部分转移在pH为3.75的乙酸钠和乙酸的溶液中并轻轻混合,以终止该部分反应(终止孔)。在添加反应溶液的一部分至终止孔之后,终止孔和反应孔应当包含相同体积的溶液。终止孔和反应孔二者在室温下培养10分钟。然后测量反应孔和终止孔在600nm处的吸光度。抑制率可以使用以下公式来计算:抑制剂(或对照)活性=(在600nm处的抑制剂终止孔的吸光度-在600nm处的抑制剂反应孔的吸光度);初始活性=在600nm处的对照酶吸光度;抑制百分数=[(初始活性/抑制剂活性)×100]-100。
过氧化物酶体增殖剂-激活的受体γ(PPAR-γ)的活性:可测定由于本说明书中所公开的活性成分、成分组合的任一种或具有该组合的组合物的每一种而引起的PPAR-γ活性变化。PPAR-γ是对皮脂产生来说重要的受体。作为一个非限制性实例,可使用分析测试的化合物或组合物抑制配体结合的能力的生物测试测定PPAR-γ的活性。简略来说,使用购自Life Technologies的荧光小分子Pan-PPAR配体FLUORMONETM Pan-PPAR Green(PV4894)来确定测试的化合物或组合物是否能够抑制配体结合至PPAR-γ。样品孔包括PPAR-γ和荧光配体,以及测试化合物或组合物(测试);参考抑制剂;罗格列酮(阳性对照);或者无测试化合物(阴性对照)。孔被培养设定的一段时间以使得配体有机会与PPAR-γ结合。然后可测定每个试样孔的荧光偏振,并与阴性对照孔比对以确定测试化合物或组合物的抑制百分比。
细胞因子阵列:人类表皮角化细胞被培养至70%至80%的融合度。吸出平板中的培养基并添加0.025%胰蛋白酶/EDTA。当细胞变得丰满时,轻轻地敲打培养皿以释放细胞。从培养皿中除去含有胰蛋白酶/EDTA的细胞并中和。将细胞在180×g下离心5分钟以形成细胞团。吸出上清液。在EPILIFETM培养基(Cascade Biologics)中再悬浮获得的细胞团。在大约10%至20%融合时将细胞接种至6孔板。在细胞变为约80%融合度之后吸出培养基,将1.0ml的EPILIFETM、佛波醇13-十四酸酯12-乙酸酯(“PMA”)(已知的炎症诱导剂)和测试组合物稀释物加入到两个重复的孔(即1.0%(100μl的100×储备液)和0.1%(10μl的100×储备液)的测试组合物稀释为终体积1ml的EpiLife生长培养基中)。培养基被轻柔地涡旋以保证充分的混合。另外,在存在或不存在额外的PMA的情况下,向对照孔中添加1.0ml的EPILIFETM。在定量给料之后,将板在37±1℃和5.0±1%CO2中培养约5小时。培育5小时后,将所有培养基收集在锥形管中并在-70℃冷冻。
为了分析,将16-垫杂交腔室连接到16-垫FAST载玻片上,该载玻片一式三份,具有16种抗细胞因子抗体和实验对照(Whatman BioSciences),并将载玻片置于FASTFrame(每帧4个载玻片)中进行处理。在室温下,使用70ml S&S蛋白质阵列阻断缓冲剂(WhatmanSchleicher and Scheull)将阵列阻断15分钟。移除阻断缓冲剂,并向每个阵列添加70ml各自的上清液试样。在轻微振动下,阵列在室温下培养3小时。使用TBS-T将阵列清洗3次。使用70ml混合型抗体处理阵列,混合型抗体含有一种对应于每个阵列捕捉生物素化抗体的抗体。在轻微振动下,阵列在室温下孵育1小时。使用TBS-T将阵列清洗3次。在轻微振动下,在室温下使用70ml含有链霉亲和素-Cy5缀合物的溶液培养1小时。使用TBS-T将阵列清洗3次,在去离子水中快速冲洗并干燥。
载玻片可以在Perkin-Elmer ScanArray 4000共聚焦荧光成像系统中成像。使用Imaging Research ArrayVision软件可以保存并分析阵列图像。简言之,点强度通过减去背景信号来确定。来自每个样品条件的点重复值可以被平均,然后与适合的对照对比。
内皮管形成:内皮管的形成和血管生成及微血管毛细血管形成有关。毛细血管形成和血管生成可以导致皮肤发红和酒槽鼻。在存在或不存在测试提取物和化合物的情况下,在细胞培养系统中,使用具有预成型的初代人脐静脉内皮细胞(HUVEC)的毛细管破坏测试可以确定内皮细胞形成管的能力。
简略来说,将HUVEC在细胞外基质进行体外培养,其刺激连接物和内皮细胞的管状形态生成以形成毛细管状的管腔结构。这些体外形成的毛细血管在许多方面与人血管的毛细血管相似。毛细管测试是基于该现象的,并用于评价潜在的脉管系统靶向试剂。
在5%CO2、37℃下,HUVEC培养物在细胞培养器中生长。用于HUVEC的完整生长基为内皮管细胞基础培养基(EBM),其补充有2%的牛胎儿血清(FBS)、12μg/ml的牛脑提取物、1μg/ml的皮质醇和1μg/ml的GA-1000(庆大霉素-两性霉素B)。在第3代至第8代之间的HUVEC培养物可以用于所有的分析测试。
用荧光试剂钙黄绿素AM预标记HUVEC,并将其接种于使用其完全生长培养基的细胞外基质涂覆的96孔培养板中。在形态发生过程后约四小时,形成内皮毛细管。然后,作为处理条件,对形成的毛细管培养物施用50μl体积的设定剂量的测试试剂。可以向未处理对照组加入测试试剂的载剂。可以包括一种浓度的FDA批准的抗血管生成药物舒尼替尼(Sutent)作为测定性能的对照。在处理后约六小时,通过显微镜检查每个孔中的内皮小管形态、使其成像,可以定量分析处理条件下的毛细管破坏活性。每个测试条件都可以在两个重复的孔中进行,包括对照组。
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本文所公开和要求保护的所有组合物和/或方法可以根据本公开可以不需要过度实验即可做出和实施。尽管本发明的组合物和方法已经按照优选的实施方案进行了描述,但是对于本领域技术人员明显的是,可以对所述组合物和/或方法以及在本文所描述方法的步骤或步骤的顺序中实施变化,而不脱离本发明的构思、精神和范围。更具体地,明显地,化学和生理两方面都相关的特定试剂可以替代本文所描述的试剂,同时会实现相同或相似的结果。所有对本领域技术人员明显的这类相似的替代和改变都被视为在如由所附权利要求限定的本发明的精神、范围和构思内。
Claims (21)
1.一种局部皮肤用组合物,其包含:
至少一种能够处理皮肤状况的皮肤活性成分;和
能够增加皮肤活性成分渗透到皮肤中的多种皮肤渗透促进剂,其中多种皮肤渗透促进剂包括:
乙醇;
异山梨醇二甲基醚;
戊二醇;
乙氧基二甘醇;和
丙二醇。
2.根据权利要求1所述的局部皮肤用组合物,其中至少一种皮肤活性成分能够减少人面部肌肉的肌肉收缩和/或减轻人皮肤上细纹或皱纹的外观。
3.根据权利要求2所述的组合物,其中至少一种皮肤活性成分是千日菊提取物、法国薰衣草提取物或迷迭香叶提取物。
4.根据权利要求3所述的组合物,其中组合物包含千日菊提取物、法国薰衣草提取物和迷迭香叶提取物。
5.根据权利要求4所述的组合物,其中组合物包含0.02重量%至2重量%的千日菊提取物、0.01重量%至1重量%的法国薰衣草提取物和0.005重量%至0.5重量%的迷迭香叶提取物。
6.根据权利要求3所述的组合物,其中每种提取物分别为水提取物、乙醇提取物、多元醇提取物或其组合。
7.根据权利要求3所述的组合物,其中:
迷迭香叶提取物是用包含甜菜碱、乳酸和水的流体提取溶剂混合物获得的;
法国薰衣草提取物是用超临界二氧化碳(CO2)提取溶剂获得的法国薰衣草花/叶/茎提取物;和
千日菊提取物是用包含水、乙醇和1,3丙二醇的流体提取溶剂混合物获得的。
8.根据权利要求1所述的组合物,其中组合物包含:
5重量%至15重量%的乙醇;
5重量%至15重量%的异山梨醇二甲基醚;
3重量%至7重量%的戊二醇;
1重量%至5重量%的乙氧基二甘醇;和
1重量%至5重量%的丙二醇。
9.根据权利要求1所述的组合物,其中组合物包含30重量%至60重量%的水。
10.根据权利要求1所述的组合物,其中组合物还包含:
鲸蜡酯;
硬脂醇;
鲸蜡硬脂醇橄榄油酸酯;
失水山梨醇橄榄油酸酯;
丙烯酸酯类/C10-30烷醇丙烯酸酯交联聚合物;
鲸蜡醇棕榈酸酯;和
失水山梨醇棕榈酸酯。
11.根据权利要求10所述的组合物,其中组合物还包含:
3重量%至7重量%的鲸蜡酯;
1重量%至5重量%的硬脂醇;
1重量%至5重量%的鲸蜡硬脂醇橄榄油酸酯;
1重量%至5重量%的失水山梨醇橄榄油酸酯;
0.1重量%至1重量%的丙烯酸酯类/C10-30烷醇丙烯酸酯交联聚合物;
0.1重量%至1重量%的鲸蜡醇棕榈酸酯;和
0.1重量%至1重量%的失水山梨醇棕榈酸酯。
12.根据权利要求1所述的组合物,其中组合物为乳液。
13.根据权利要求12所述的组合物,其中乳液为水包油乳液。
14.一种改善人的皮肤外观的方法,该方法包括将根据权利要求1所述的组合物局部施用至人的皮肤上,其中该组合物改善了皮肤的外观。
15.根据权利要求14所述的方法,其中将组合物施用至细纹或皱纹。
16.根据权利要求15所述的方法,其中在局部施用后减轻了细纹或皱纹的外观。
17.根据权利要求16所述的方法,其中组合物减少了引起细纹或皱纹外观的肌肉的肌肉收缩。
18.根据权利要求17所述的方法,其中面部肌肉是眉间复合肌、眼轮匝肌、降肌、或额肌或其任意组合。
19.根据权利要求15所述的方法,其中细纹或皱纹是面部皱纹。
20.根据权利要求14所述的方法,其中组合物包含:
千日菊提取物,其中千日菊提取物刺激了皮肤中胶原蛋白的产生和层黏连蛋白的产生;和
迷迭香叶提取物,其中迷迭香叶提取物减少了皮肤中基质金属蛋白酶1、基质金属蛋白酶3或基质金属蛋白酶9的产生。
21.根据权利要求14所述的方法,其中面部皮肤是前额皮肤、脸颊皮肤、下巴皮肤和/或眼眶区域皮肤。
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US2798053A (en) | 1952-09-03 | 1957-07-02 | Goodrich Co B F | Carboxylic polymers |
US3755560A (en) | 1971-06-30 | 1973-08-28 | Dow Chemical Co | Nongreasy cosmetic lotions |
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KR102278274B1 (ko) * | 2014-04-30 | 2021-07-16 | 킴벌리-클라크 월드와이드, 인크. | 피부 노화의 징후를 감소시키기 위한 미역 추출물의 사용 |
EP3817755A4 (en) * | 2018-07-03 | 2022-04-20 | Mary Kay Inc. | TOPICAL MUSCLE RELAXATION COMPOSITIONS AND METHODS |
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2022
- 2022-07-01 EP EP22748205.6A patent/EP4363062A1/en active Pending
- 2022-07-01 CN CN202280058123.3A patent/CN117940103A/zh active Pending
- 2022-07-01 WO PCT/US2022/073353 patent/WO2023279086A1/en active Application Filing
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