CN102451467B - 包含类视色素和NFκB抑制剂的组合物及其使用方法 - Google Patents
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Abstract
本发明提供了一种组合物，所述组合物包含类视色素、NFκB抑制剂和美容上可接受的局部用载体。本发明还提供了处理皮肤的方法。
Description
技术领域
提供了一种包含类视色素、NFκB抑制剂和美容上可接受的局部用载体的组合物。该组合物可用于局部施用至皮肤。
背景技术
类视色素已知可治疗不同的皮肤病症如痤疮或光老化。然而，用于局部施用的类视色素(如视黄酸、视黄醛或视黄醇)可在某些情况下导致发红、瘙痒、刺痛、皮肤剥落或其它刺激表现。本发明人已考虑到降低发炎的一种可能的解决方案是在局部用组合物中使用较低浓度的类视色素。然而，可理解地，具有低水平类视色素的组合物可具有低的功效。
本发明现已惊人地发现，可将特定的一类抗炎化合物即抑制细胞转录因子核转录因子k-B(NFκB)的试剂与类视色素以一定方式组合，该方式使得视黄醇活性有惊人地大的、出人意料的协同增强，该协同增强可通过CRAPBII基因的表达来测量。
发明内容
在一个方面，本发明提供了包含类视色素、NFκB抑制剂和美容上可接受的局部用载体的组合物，其中该组合物中的类视色素的量不超过该组合物的约0.075重量％。
在另一方面，本发明提供了处理皮肤的方法，该方法包括局部施用上述组合物至哺乳动物的皮肤。
通过本发明的详细描述和权利要求书，本发明的其他特征和优点将变得显而易见。
具体实施方式
据信，本领域的技术人员可以根据本文的描述最大程度地利用本发明。以下具体实施例应被理解为仅仅是说明性的，而不以任何方式限制本公开内容的其余部分。
除非另有规定，否则本文使用的所有技术和科学术语均具有本发明所属技术领域普通技术人员公知的相同含义。除非另外指明，否则量、百分比或浓度指以重量计的量、百分比或浓度(即％(W/W))。除非另外阐明，否则所有范围均包括端值，如“4至9”包括端值4和9。
如本文所用，“局部施用”意指例如用手或涂敷器(例如擦拭物、滚筒或喷雾器)直接涂在或铺展在外部皮肤、头皮或毛发上。
如本文所用，“美容上可接受的”意指该术语描述的成分适于与组织(如，皮肤或毛发)接触使用而不会引起不适当的毒性、不相容性、不稳定性、剌激性、变应性反应等。
本文所用的术语“治疗”或“处理”意指治疗(如症状的缓或消除和/或治愈)和/或预防或抑制病症(如皮肤病症)。
本发明的组合物适用于处理哺乳动物皮肤，例如用于改善多种皮肤老化迹象，例如紧实度(firmness)、肌理(texture)或皱纹出现。如本文所用，“需要改善老化迹象的皮肤”意指为(但不限于)松垂、松弛、松散、粗糙、有皱纹、变薄或不平的皮肤。改善老化迹象意指改善皮肤紧实度、改善皮肤肌理、改善皮肤中细纹或皱纹的出现、改善皮肤色调(tone)或处理皮肤中的外部侵害。
如本文所用，“改善皮肤的紧实度”意指增强皮肤的紧实度或弹性，预防皮肤紧实度或弹性损失或者预防或处理松垂、松散和松弛的皮肤。皮肤的紧实度或弹性可通过使用皮肤弹性测试仪(cutometer)测量。参见Handbook of Non-Invasive Methods and the Skin，J.Serup，G.Jemec&G.Grove(编辑)，第66.1章(2006)。皮肤弹性或紧实度的损失可能是多种因素造成的，包括但不限于老化、疾病、激素变化、创伤、环境损害或施用化妆品至皮肤的结果。
如本文所用，“改善皮肤的肌理”意指使皮肤表面平滑以除去皮肤表面上的隆起块或裂隙。
如本文所用，“改善皮肤中细纹或皱纹的出现”意指预防、延缓、停止或逆转皮肤中皱纹和细纹形成的过程。
如本文所用，“处理皮肤中的外部侵害”意指减少或预防皮肤中外部侵害带来的损害。外部侵害的例子包括但不限于使用清洁剂(例如含有表面活性剂的局部用清洁剂)、化妆品、剃须以及环境损害如来自UV光照(例如来自日光的阳光损害或来自非天然来源如UV灯和太阳模拟器的损害)、臭氧、废气、污染、氯和含氯化合物和抽烟带来对皮肤的损害。外部侵害对皮肤的影响包括但不限于对脂质、碳水化合物、肽、蛋白质、核酸和维生素的氧化和/或亚硝化损伤和对这些物质的修饰。外部侵害对皮肤的影响还包括但不限于细胞活力的损失、细胞功能的损失或改变以及基因和/或蛋白质表达的变化。
如本文所用，术语“美白皮肤”通常指使皮肤色调、皮肤颜色和/或皮肤色度(shade)变淡、变亮、变白和/或均匀，和/或指祛黄，和/或指色素斑痕和/或病损(包括但不限于色素斑、黑色素斑、老年斑、日晒斑、衰老性着色班、雀斑(freckles)、单纯性雀斑痣、色素性日光性角化病、脂溢性角化病、黑斑病、痤疮斑痕、发炎后色素沉着过度、着色斑、雀斑(ephelides)、它们中两种或更多种的组合等等)的淡化和/或褪色。在某些实施例中，“美白皮肤”还指增加的皮肤光亮度、光泽度、半透明度和/或发光和/或获得更光亮、光泽、半透明的或发光的皮肤色调外观或较低的黄色或萎黄色皮肤色调。在某些优选的实施例中，“美白皮肤”指使皮肤色调变淡和均匀，增加皮肤光亮度和/或淡化老年斑。
如本文所用，术语“需要进行皮肤美白处理的皮肤”一般指表现出一种或多种选自如下的特性的皮肤：具有低于41的个体类型角(IndividualTypology Angle，ITA)测量值(根据2007年公布的COLIPA GUIDELINE：GUIDELINE FOR THE COLORIMETRIC DETERMINATION OF SKINCOLOUR TYPING AND PREDICTION OF THE MINIMAL ERYTHEMALDOSE(MED)WITHOUT UV EXPOSURE测定，将该文献以引用的方式并入本文，并且其在下文进一步描述)的皮肤，发暗和/或萎黄的皮肤，包括UV暗化的皮肤，具有不均匀皮肤色调的皮肤，或具有一处或多处色素斑痕和/或病损(包括但不限于色斑、黑色素斑、老年斑、日晒斑、衰老性着色班、雀斑(freckles)、单纯性雀斑痣、色素性日光性角化病、脂溢性角化病、黑斑病、痤疮斑痕、发炎后色素沉着过度、着色斑、雀斑(ephelides)它们中两种或更多种的组合等等)的皮肤。在COLIPA指南中，皮肤颜色根据ITA值定义为：十分浅色的皮肤＞55；浅色皮肤41-55，中等色皮肤28-41，棕褐色皮肤＜28。在某些优选的实施例中，“需要进行皮肤美白的皮肤”指皮肤的ITA值低于41，例如约40或更低，约35或更低，约30或更低，或更优选约28或更低的个体。在某些优选的实施例中，本发明涉及用于选自萎黄和/或暗化的皮肤的需要进行皮肤美白处理的皮肤上的组合物和方法。在某些其他优选的实施例中，本发明涉及用于需要进行选自老年斑、雀斑、痤疮后留下的斑痕以及它们中两种或更多种的组合的皮肤美白处理的组合物和方法。
本发明的组合物还适于处理痤疮。如本文所用，“处理痤疮”指减轻、减少、预防、改善或消除由激素和其他物质对皮脂腺及其毛囊的作用(通常在皮肤上导致毛孔阻塞和形成炎性或非炎性病损)而引起的障碍的存在或迹象。具体而言，其涉及治疗或预防疵点、病损或丘疹、出现前丘疹、黑头和/或白头。如本文所用，“出现前丘疹”是发炎的毛囊，这些毛囊在皮肤表面上用肉眼观察时不明显(如作为病损)。
如本文所用，“化妆品”指能保持、恢复、赋予、刺激或增强体美外观或者似乎能增进美貌或年轻的美容物质或制品，特别是涉及组织或皮肤的外观时。
如本文所用，“美容有效量”意指足以治疗皮肤老化或痤疮的一种或多种迹象，但低得足以避免严重副作用的生理活性化合物或组合物的量。化合物或组合物的美容上有效量将会随以下因素而变：所治疗的具体病症、最终使用者的年龄和身体状况、所治疗/预防的病症的严重程度、治疗的持续时间、其他治疗的性质、所采用的具体化合物或产品/组合物、所具体采用的美容上可接受的载体等等。
类视色素
本发明的组合物包含至少一种类视色素。如本文所用，“类视色素”意指来自一类与维生素A结构类似的化合物的化合物，如由如下结构表征的那些化合物：
其中R代表官能团如CH2OH(视黄醇)、CHO(视黄醛)、CO2H(视黄酸)、CH2OCOCH(视黄醇乙酸酯)。其他衍生物也是合适的，例如诸如视黄醇棕榈酸酯之类的其他酯、胺衍生物等等。在一个实施例中，类视色素选自视黄醇、视黄醛、视黄酸、视黄醇乙酸酯和视黄醇棕榈酸酯。在一个优选的实施例中，类视色素是视黄醇。
本发明人发现，可在组合物中维持相对低水平的类视色素水平，却仍提供比预期功效惊人地大的功效。在本发明的某些实施例中，类视色素(例如视黄醇)在组合物中的量不超过组合物的约0.075重量％(例如，不超过约0.0026摩尔/升组合物，或不超过约247,500维生素A国际单位/100克组合物)。在某些实施例中，类视色素在组合物中的量为组合物的约0.01重量％至约0.075重量％(例如，约0.000349至约0.0026摩尔/升组合物)，如约组合物的约0.01重量％至约0.06重量％(例如，约0.000349至约0.00209摩尔/升组合物)。在一个优选的实施例中，组合物包含处于上述浓度范围内的视黄醇。
NFκB抑制剂
本发明的组合物包含至少一种NFκB抑制剂。如本文所用，“NFκB抑制剂”意指可抑制细胞转录因子核转录因子k-B(NFκB)的化合物。在一个实施例中，NFκB抑制剂在根据下文定义的“NFκB抑制测试”进行测试时，在以优选每毫升1微克至约每毫升100微克的浓度测试时具有至少约35％、优选至少约55％、更优选至少约70％、最优选至少约90％的NFκB抑制百分比。也就是说，化合物以在每毫升1微克至每毫升100微克范围内的至少一种浓度下显示出所列出的NFκB抑制百分比。该化合物不必在每毫升1毫克至每毫升100毫克的所有浓度下提供所列出的NFκB抑制百分比，而是在该范围内的至少一种浓度下提供所列出的NFκB抑制百分比。
在一个优选的实施例中，当以每毫升10毫克的浓度测试时，NFκB抑制剂具有至少约35％、优选至少约55％、更优选至少约70％、最优选至少约90％的NFκB抑制百分比。
“NFκB抑制测试”以如下方式进行。使用FB293细胞，其为含有NF-kB的报告基因的稳定转染的人上皮细胞系。它们可从例如Panomics(Fremont，CA)获得。将FB293以5×104个细胞/mL的密度接种于合适的培养基中，例如补充有10％胎牛血清(Invitrogen，San Diego，CA)的达尔伯克氏改良伊格尔培养基(DMEM)中。在存在测试样品的情况下用100ng/mL的肿瘤坏死因子α(TNFα，可得自Sigma-Alrich(St Louis，MO))刺激FB293细胞。独立地，测试了对照样品，其中未施加测试样品。在37℃下使用5％CO2温育24小时后，将细胞用40μl的报告基因裂解缓冲液(Promega，Madison，WI)裂解。将20μL的裂解产物等分试样用虫荧光素酶测定试剂盒(Promega)进行测定，并在Lmax发光计(Molecular Devices，Sunnyvale，CA)中计数10秒，数据以相对光单位/秒表示。测试样品的NFκB抑制百分比计算如下：
NFκB抑制＝[1-(L样品/L对照)]*100
其中L样品为样品的发光，L对照为对照的发光。
在一个实施例中，NFκB抑制剂选自如下化合物：取代的间苯二酚、(E)-3-(4-甲基苯基磺酰基)-2-丙烯腈(如“Bay 11-7082”，可从Sigma-Aldrich(St.Louis，Missouri)商购获得)、四氢姜黄素(如四氢姜黄素CG，可得自Sabinsa Corporation(Piscataway，NJ))、锈毛泡桐(Paulowniatomentosa)木的提取物以及它们的组合。在另一个实施例中，NFκB抑制剂选自取代的间苯二酚、四氢姜黄素以及它们的组合。
在一个实施例中，NFκB抑制剂包含锈毛泡桐木的提取物。泡桐属(Paulownia)是亚洲原产的一个植物属，其已逐渐传播至欧洲和美国。在日本，泡桐属被称为kiri，其具体地指一个物种锈毛泡桐，也称为“公主树”。通常使用的其他名字是“皇后树”、“毛地黄树”、“皇家泡桐”、“泡桐”(中文名)和“Odong-Namoo”(朝鲜名)。学名为“锈毛泡桐”，在多篇文献中报道了多个别名，即“Paulownia imperialis”、“Paulownia recurva”和“Bignonia tomentosa”。锈毛泡桐属于“泡桐科(Paulowniaceae)”或有时称为“玄参科(Scrophulariaceae)”。美国农业部(plants.USDA.gov)植物数据库通过唯一符号“PATO2”标识皇后树，以锈毛泡桐和Paulownia imperialis作为别名。
可使用任何合适的锈毛泡桐木的提取物。一般而言，锈毛泡桐树的木材包括来自树杆、树枝或这二者的组合的木材。锈毛泡桐木的合适提取物可源于木屑、木粉和/或小切屑等等。
锈毛泡桐木的合适提取物可用常规的方法获得，包括但不限于：通过研磨、浸渍、压榨、挤压、捣碎、离心，和/或诸如冷渗漉、搅拌/蒸馏、微波辅助萃取、超声处理、使用或不使用极性修饰剂的超临界/亚临界CO2压缩气体提取、加压溶剂提取、加速溶剂提取、加压或常压热水提取、表面活性剂辅助的加压热水提取、油提取、膜提取、索氏提取、例如在颁给Integrated Botanical Technologies，LLC的第7442391号、第7473435号和第7537791号美国专利中公开(以引用的方式并入本文)的金手指蒸馏/提取(gold finger distillation/extraction)和/或工艺等等之类的工艺，或通过诸如溶剂萃取等之类的其他方法从木材直接提取物质。包括极性溶剂、非极性溶剂或它们中的两种或更多种的组合在内的多种溶剂中的任一种可用于包括溶剂萃取的方法中。合适的极性溶剂包括：极性无机溶剂如水等等，极性有机溶剂如一元醇和相应的有机酸(例如包括甲醇、乙醇、丙醇、丁醇等等在内的C1-C8醇和包括乙酸、甲酸、丙酸等等在内的有机酸)、多元醇和二醇(包括C1-C8多元醇/二醇等等)以及它们两种或更多种的组合。合适的非极性溶剂包括非极性有机溶剂如烷烃(包括C1-C8烷烃)、环烷烃(包括C1-C8烷烃)、烷基醚(包括C1-C8烷基醚)、石油醚、酮(包括C1-C8酮)、二氯甲烷、乙酸乙酯、二甲苯、甲苯、氯仿、植物油、矿物油等等。在另一个实施例中，可通过上述非极性溶剂或者使用或不使用极性修饰剂如(C1-C8醇、水、C1-C8多元醇/二醇或C1-C8有机酸)的超临界流体提取而获得提取物。在某些优选的实施例中，本发明的提取物是通过将木材研磨成粉并用20℃下介电常数值在1至100之间的极性溶剂提取，优选用20℃下介电常数值在4至60之间的极性溶剂提取，更优选用20℃下介电常数值在4和50之间的极性溶剂提取，还更优选用20℃下介电常数在4至40之间的极性溶剂提取而制备的极性提取物。优选的极性溶剂的例子包括20℃下介电常数值在1至100之间，优选在4至60之间，更优选在5至40之间的C1-C8醇、C1-C8多元醇/二醇、C1-C8有机酸、水以及它们两种或更多种的组合，包括(但不限于)如“Dielectric Constants of SomeOrganic Solvent-Water Mixtures at Various Temperatures，”Akerlof，Gosta；JACS，第54卷，第11期(1932年11月)，第4125-4139页(以引用的方式并入本文)中所述的具有所需的介电常数值的那些溶剂及溶剂的组合。在某些优选的实施例中，用一种或多种C1-C8一元醇、C1-C8多元醇、C1-C8二醇以及它们中两种或更多种的组合提取极性提取物。在某些更优选的实施例中，用一种或多种C1-C4一元醇、C1-C4多元醇和/或C1-C4二醇提取提取物。在某些更优选的实施例中，在存在或不存在水的情况下用包含甲醇、乙醇或它们的组合的溶剂制备提取物。在更优选的实施例中，用无水醇或试剂级变性醇和干Kiri木材粉，在室温下搅拌3天来制备提取物。在某些优选的实施例中，可通过木炭(也称为活性炭)进一步精制提取物。
在某些实施例中，可将锈毛泡桐提取物制备为基本上不含某些物质。在一个实施例中，提取物基本上不含乌索酸、β-谷甾醇或这二者。
在某些实施例中，组合物可另外包含来自锈毛泡桐的其他部分(例如树皮、树叶、根、果实、种子或花中的一者或多者)的提取物。在其它实施例中，组合物基本上不含来自锈毛泡桐的其他非木质部分的提取物。
在一个优选的实施例中，NFκB抑制剂是取代的间苯二酚。间苯二酚是具有如下结构的二羟基酚化合物(即1，3二羟基苯)：
如本文所用，“取代的间苯二酚”意指在2、4、5或6位中具有至少一个取代基间苯二酚。因而，取代的间苯二酚可具有少至一个且多至四个的取代基。取代的间苯二酚的位置1和3具有-OH羟基，如上所示。
高度优选的是，取代的间苯二酚的所有取代基无苯基(-C6H5芳族)部分。在某些实施例中，所有取代基无(具有或不具有杂原子的)芳族部分。
在另一个实施例中，优选的是取代的间苯二酚的所有取代基无酮官能团(与另外两个碳原子键合的羰基)。
在某些优选的实施例中，取代的间苯二酚的所有取代基既无苯基官能团也无酮官能团。
在某些优选的实施例中，取代的间苯二酚包含至少一个具有5至11个碳原子、优选5至10个碳原子、更优选5至9个碳原子、最优选5至8个碳原子的取代基。在某些其他实施例中，至少一个取代基包含烷基，例如具有上述碳原子数目的烷基。该烷基优选是不饱和的。
在某些实施例中，间苯二酚的4位被取代，在某些实施例中，仅4位被取代。在另一个实施例中，4位置被烷基基团取代。在某些优选的实施例中，取代的间苯二酚包含单个4位取代基，该取代基包含烷基。在某些其他的优选实施例中，取代的间苯二酚包含单个4位取代基，该取代基由与苯环直接键合的烷基组成。
特别合适的取代的间苯二酚包括4-己基间苯二酚和4-辛基间苯二酚，特别是4-己基间苯二酚。4-己基间苯二酚和4-辛基间苯二酚的结构在下面示出：
4-己基间苯二酚
4-辛基间苯二酚
4-己基间苯二酚可以商品名“SYNOVEA HR”购自Sytheon(Lincoln Park，NJ)。4-辛基间苯二酚可购自City Chemical LLC(West Haven，Connecticut)。
在某些实施例中，取代的间苯二酚在2、4、5或6位具有至少两个取代基。这种取代基可任选连接而形成环，例如任选包含杂原子例如硫或氧的环状脂族烃。例如连接的取代基可包含5至10碳原子，如8至10碳原子，并任选包括1至3杂原子。具有连接2位和3位的环状脂族取代基的合适取代的间苯二酚的例子包括玉米赤霉酮和β-玉米赤霉醇：
玉米赤霉酮
β-玉米赤霉醇
玉米赤霉酮和β-玉米赤霉醇可从Sigma Chemicals(St.Louis，Missouri)商购获得。
在某些其他实施例中，取代的间苯二酚具有含卤和/或含亚硝基的取代基。合适的例子含有与苯环直接键合的-Cl或-N＝O。这些取代基可存在于(例如)2和4位、2和6位或4和6位上。二卤取代的间苯二酚的例子为2，6-氯间苯二酚。二亚硝基取代的间苯二酚的例子为2，4-二亚硝基间苯二酚：
2，4-二亚硝基间苯二酚
2，6-二氯间苯二酚和2，4-二亚硝基间苯二酚可得自City Chemical LLC(WestHaven，Connecticut)。
取代的间苯二酚可通过本领域已知的方法制备，例如使用美国专利No.4,337,370中描述的技术制备，将该专利的内容以引用的方式并入本文。
取代的间苯二酚可具有任何合适的分子量。在某些实施例中，取代的间苯二酚的分子量在约175至约300之间的范围内。
CRAPBII表达
在本发明的一个实施例中，对组合物中的NFκB抑制剂的浓度进行选择以(与类视色素联合)使CRAPBII表达有至少约15％增效，CRAPBII表达可通过下文所述的“CRAPBII表达测试”来测量。这种组合有利地使得能使用相对低水平的类视色素(其有时候是刺激性的)，而同时保持高度的类视色素活性。在人中，CPABPII基因编码细胞视黄酸结合蛋白的合成。就这一点而论，CRABPII基因的表达直接涉及类视色素功效并强烈与之相关。本发明人已发现，在含有类视色素的组合物中存在NFκB抑制剂可改善或强化类视色素的有益效果，这可通过CRAPB II的表达来测量。
类视色素和NFκB抑制剂的特定组合的以CRAPBII表达示出的增效程度(“％增效”)可用“CRAPBII表达测试”测定。“CRAPBII表达测试”以如下方式进行。在整形手术后从皮肤腹部活检样本制备一厘米直径的离体皮肤外植体。将皮肤外植体在水饱和的大气环境中在37℃下维持在补充有0.125μg/ml两性霉素B的KGM goldTM培养基中，测试持续时间为48小时。将外植体置于常规的测试板中，表皮表面向上，并且处于足够的培养基中以几乎但非完全浸没样品(即表皮表面从培养基的上表面突出)。将测试组合物施加至从培养基突出的表皮表面。48小时后，使用本领域技术人员已知的常规技术，将外植体移出，从其提取表皮mRNA，利用合适的寡核苷酸序列通过定量实时PCR(QRT-PCR)测量CPABPII基因的表达。
将含有所需类视色素的油包水乳液基质用于测试组合物。NFκB抑制剂可任选包括在测试组合物中或单独混合进培养基中。例如，取决于NFκB抑制剂的相对渗透速率，可能有利的是将其包括在测试组合物或培养基中以消除作为测试中的可变因素的渗透速率。如果包含在培养基中，NFκB抑制剂不需要从突出的表皮表面渗透进表皮中，而是可能从底下的培养基渗透进表皮中。如果NFκB抑制剂包括在测试组合物中，则NFκB抑制剂必须从突出的表皮表面渗透进表皮中。
可调节测试组合物中水的浓度以适应足够水平的视黄醇(以及NFκB抑制剂，如果存在于测试组合物中的话)。可用于测试组合物中的水包油乳液基质的例子在下表1中示出：
表1：用于CRAPBII表达测试的合适水包油乳液基质
| 商品名 | INCI | 浓度(重量百分比) |
| 去矿物质水 | 水 | 83.4 |
| Carbopol Ultrez 10 | 卡波姆 | 0.4 |
| Edeta BD螯合剂 | 乙二胺四乙酸二钠 | 0.1 |
| Nipagin M | 对羟基苯甲酸甲酯 | 0.2 |
| 对羟基苯甲酸丙酯 | 对羟基苯甲酸丙酯 | 0.15 |
| 苯氧基乙醇 | 苯氧基乙醇 | 0.5 |
| 氢氧化钠 | 氢氧化钠 | 0.1 |
| Simulsol 165 | PEG-100硬脂酸酯；硬脂酸甘油酯 | 2.0 |
| Nipanox BHT | BHT | 0.1 |
| Lanette 16 | 鲸蜡醇；硬脂醇；十四醇 | 1.0 |
| DUB ININ | 异壬酸异壬酯 | 7.0 |
| 丙二醇 | 丙二醇 | 5.0 |
| 抗坏血酸晶体 | 抗坏血酸 | 0.05 |
(i)单独针对给定量的类视色素，(ii)单独针对给定量的NFκB抑制剂，以及(iii)针对量(i)的类视色素和量(ii)的NFκB抑制剂的混合物，测定CRAPBII表达。用如下公式计算％增效：
％增效＝100％×[[CRAPBII混合物/(CRAPBII类视色素+CRAPBIINFκB抑制剂)]-1]
虽然可选择NFκB抑制剂的浓度来为本发明组合物提供至少约15％的增效CRABPII表达，但在某些实施例中，可选择NFκB抑制剂的水平以在CRABPII表达方面提供更高水平的增效，例如至少约50％，或至少约80％的增效。
在一个实施例中，可例如，利用“NFκB抑制测试”的结果选择NFκB抑制剂的浓度。例如，可通过以若干(例如4种或更多种)浓度进行“NFκB抑制测试”来估计合适的浓度，如下面实例1中所述。可选择NFκB抑制剂的浓度，例如，当以10微克/毫升的浓度测试时使得可提供至少约35％、优选至少约55％、更优选至少约70％、最优选至少约90％的NF-kB抑制百分比。可将该浓度的NFκB抑制剂与不超过约0.075重量％的类视色素在组合物中组合并测定CRAPBII表达的％增效，如下面实例2中所示。
本发明人已观察到，在某些实施例中，不仅在低浓度的类视色素(不超过约0.075重量％)下，而且在低浓度的NFκB抑制剂下都可惊人地观察到CRAPBII表达的增效。这是有利的，因为类视色素的浓度可维持在低的水平，从而避免了类视色素可能的负面刺激副作用，而同时从NFκB抑制剂获得强效的功效增强。
在本发明的某些实施例中，组合物包含不超过约0.075重量％的类视色素，例如视黄醇，以及不超过约0.5重量％的NFκB抑制剂。在一个优选的实施例中，组合物包含不超过约0.075％的类视色素和约0.1％至约0.5％的NFκB抑制剂。
局部用组合物
本发明的组合物可局部施用至人皮肤和/或毛发。除了NFκB抑制剂和类视色素外，组合物还包含美容上可接受的局部用载体，该局部用载体可为组合物的约50重量％至约99.99重量％(如组合物的约80重量％至约99.9重量％)。在本发明的优选实施例中，美容上可接受的局部用载体是水或包含水。美容上可接受的局部用载体可包括一种或多种选自润湿剂、润肤剂、油脂、湿润剂等等。在一个实施例中，美容上可接受的局部用载体包括诸如无纺布或膜材料之类的基材。
可将组合物制备成多种产品类型，包括但不限于洗剂、霜剂、凝胶剂、棒剂、喷雾剂、油膏剂、清洁液体洗剂和固体皂剂、洗发剂、糊剂、泡沫剂、粉末剂、摩丝、剃须膏、擦拭物、贴剂、水凝胶、成膜产品、面膜和皮肤膜剂、膜剂和化妆品如粉底以及睫毛膏。这些产品类型可含有几种类型的美容上可接受的局部用载体，包括(但不限于)溶液、悬浮液、乳状液(如微乳液和纳米乳液)、凝胶、固体和脂质体。
可用于本发明的组合物能配制成溶液。溶液剂通常包含水性溶剂或有机溶剂(例如，约50％至约99.99％或约90％至约99％的美容上可接受的水性溶剂或有机溶剂)。合适的有机溶剂的例子包括：丙二醇、聚乙二醇(200-600)、聚丙二醇(425-2025)、甘油、1，2，4-丁三醇、山梨醇酯、1，2，6-己三醇、乙醇以及它们的混合物。
可用于本主题发明的组合物可配制成包含润肤剂的溶液。此类组合物优选包含约2％至约50％的一种或多种润肤剂。如本文所用，“润肤剂”是指用于例如通过预防水分经表皮从皮肤流失来预防或缓解干燥的材料。润肤剂的例子包括但不限于植物油、矿物油、脂族酯等。
洗剂可以由此类溶液制成。洗剂通常包含约1％至约20％(例如，约5％至约10％)的一种或多种润肤剂和约50％至约90％(例如，约60％至约80％)的水分。
可由溶液配制的另一种产品是霜剂。霜剂通常包含约5％至约50％(例如，约10％至约20％)的一种或多种润肤剂和约45％至约85％(例如，约50％至约75％)的水分。
尽管优选的是本发明的组合物包括水，但是作为另一种选择，组合物可以是无水的或者是不含水但含有机和/或硅树脂溶剂、油、类脂和蜡的膏剂。膏剂可包含简单的动物或植物油基质或半固体碳氢化合物。膏剂可包含约2％至约10％的一种或多种润肤剂和约0.1％至约2％的一种或多种增稠剂。
组合物可以配制为乳状液。如果局部用载体是乳状液，则约1％至约10％(如，约2％至约5％)的局部用载体包含乳化剂。乳化剂可以为非离子型、阴离子型或阳离子型。合适的乳化剂的例子包括个人护理和化妆品配方领域中通常鉴定为合适乳化剂的那些。
洗剂和软膏可配制成乳剂。通常此类乳液包含0.5％至约5％的乳化剂。这种霜剂通常含有约1％至约20％(如约5％至约10％)的一种或多种润肤剂；约20％至约80％(如30％至约70％)的水；以及约1％至约10％(如约2％至约5％)的一种或多种乳化剂。
水包油和油包水型单乳液护肤制剂，例如洗剂和软膏是化妆品领域所熟知的，并且可用于本主题发明。多相乳状液组合物(例如水包油包水型或油包水包油型)也可用于本主题发明。通常，此类单相乳状液或多相乳状液含有水分、润肤剂和乳化剂作为其基本成分。
本发明的组合物还可配制成凝胶(例如，使用合适的胶凝剂制成的水性凝胶、醇类凝胶、醇/水型凝胶或油性凝胶)。用于水性凝胶和/或醇类凝胶的适当胶凝剂包括但不限于天然树胶、丙烯酸和丙烯酸酯聚合物和共聚物以及纤维素衍生物(例如羟甲基纤维素和羟丙基纤维素)。用于油(例如矿物油)的适当胶凝剂包括但不限于氢化丁烯/乙烯/苯乙烯共聚物和氢化乙烯/丙烯/苯乙烯共聚物。此类凝胶通常包含介于约0.1重量％和5重量％之间的此类胶凝剂。
本发明的组合物也可配制成固体制剂(例如蜡基棒、条皂组合物、粉末或含有粉末的擦拭物)。
除了前述的组分外，用于本主题发明的组合物还可包含各种各样的其他油溶性物质和/或水溶性物质，这些物质通常用于组合物中，按本领域既定的水平施用于皮肤和头发。
另外的美容活性剂
在一个实施例中，组合物进一步包含另一种美容活性剂如本文所用，“美容活性剂”是对皮肤或毛发具有美容或治疗效果的化合物(如，合成化合物或从天然来源或天然提取物分离的化合物)，其包括(但不限于)抗痤疮剂、控油剂、抗微生物剂(非NFκB抑制剂)、抗真菌剂、抗寄生虫剂、外用止痛剂、防晒剂、光防护剂、抗氧化剂、角质层分离剂、表面活性剂、保湿剂、营养物质、维生素、能量增强剂、防汗剂、收敛剂、除臭剂、紧致剂、抗胼胝剂(anti-callous agent)以及用于毛发和/或皮肤调理的试剂。
在一个实施例中，所述试剂选自(但不限于)羟基酸、过氧化苯甲酰、D-泛醇、甲氧基肉桂酸辛酯、二氧化钛、水杨酸辛酯、胡莫柳酯、阿伏苯宗、类胡萝卜素、自由基清除剂、自旋捕获剂、胺(如neutrol)、神经酰胺、多不饱和脂肪酸、必需脂肪酸、酶、酶抑制剂、矿物质、诸如雌激素之类的激素、诸如氢化可的松之类的类固醇、2-二甲基氨乙醇、诸如氯化铜之类的铜盐、诸如Cu:Gly-His-Lys之类的含铜肽、辅酶Q10、肽、诸如脯氨酸之类的氨基酸、维生素、乳糖酸、乙酰辅酶A、烟酸、核黄素、硫胺素、核糖、诸如NADH和FADH2之类的电子传递物质和诸如芦荟、小白菊、燕麦片之类的其他植物提取物以及它们的衍生物和混合物。美容活性剂通常将以组合物的约0.001重量％至约20重量％(如，约0.005％至约10％，例如约0.01％至约5％)的量存在于本发明的组合物中。
维生素的例子包括但不限于维生素B(如维生素B3、维生素B5和维生素B 12)、维生素C、维生素K和维生素E的不同形式(如α、β、γ或Δ生育酚)或它们的混合物以及它们的衍生物。
羟基酸的例子包括，但不限于乙醇酸、乳酸、苹果酸、水杨酸、柠檬酸和酒石酸。
抗氧化剂的例子包括(但不限于)水溶性抗氧化剂，例如巯基化合物和它们的衍生物(如，焦亚硫酸钠和N-乙酰基-半胱氨酸)、硫辛酸和二氢硫辛酸、白藜芦醇、乳铁蛋白以及抗坏血酸和抗坏血酸衍生物(如，抗坏血酸棕榈酸酯和抗坏血酸多肽)。适用于本发明的组合物中的油溶性抗氧化剂包括(但不限于)丁基化羟基甲苯、生育酚类(例如，生育酚乙酸酯)、生育三烯酚类和泛醌。包含适用于本发明的组合物中的抗氧化剂的天然提取物包括(但不限于)包含黄酮类和异黄酮类以及它们的衍生物(如，染料木黄酮和二联玉米蛋白)的提取物、包含白藜芦醇的提取物等。此类天然提取物的例子包括葡萄籽、绿茶、松树皮和蜂胶。
其他材料
各种其他材料也可以存在于组合物中，如本领域所公知的。这包括湿润剂、pH调节剂、螯合剂(例如EDTA)、芳香剂、染料和防腐剂(例如对羟基苯甲酸脂)如化妆品配方中通常使用的那些。
包含本发明此类组合物的组合物、制剂和产品可以利用本领域普通技术人员所知的方法来制备。
使用方法
可将本发明的组合物局部施用至需要处理以改善如上所述的皮肤老化或痤疮的一种或多种迹象的哺乳动物皮肤。在一个实施例中，可将组合物施用至需要在紧实度、肌理或细纹和皱纹出现方面改善的皮肤。可根据合适的处理方案将组合物施用至需要这种处理的皮肤，例如从多达每天两次到少至每三天一次左右。
在某些实施例中，本发明的组合物还可用于处理与皮肤相关的其他需要状态。例如，本发明的组合物可用于处理发炎后色素沉着过度、用于减少毛孔大小、用于减少皮脂产生以及用于缓和疤痕。在某些其他实施例中，可将本发明的组合物与机械或物理表皮剥脱处理(例如微晶磨皮处理)同时施用或在该处理的数小时内施用，或与化学剥脱剂或角质层分离剂例如水杨酸同时施用。在某些其他实施例中，可将本发明的组合物施用至轻微的伤口或手术后部位以帮助痊愈。
据信，本领域的技术人员可以根据本文的描述最大程度地利用本发明。以下具体实施例应被解释为仅仅是说明性的，而不以任何方式限制本公开内容的其余部分。以下非限制性实例进一步说明了本发明。
实例1：NFκB抑制
对如下测试样品进行了上述“NFκB抑制测试”：Bay 11-7082测试样品(Sigma-Aldrich，St.Louis，MO)、四氢姜黄素CG(Sabinsa Corporation，Piscataway，NJ)以及多种浓度的4-己基间苯二酚。结果在表2中示出，其中对测试样品和对照样品记录NF-kB报告基因激活(发光，L)。还记录了NF-kB抑制百分比。
表2：NFκB抑制测试的结果
Bay 11-7082、四氢姜黄素CG和4-己基间苯二酚显示出强的NF-κB抑制。
实例2：4-己基间苯二酚和视黄醇-CRAPBII的表达
对一系列含有视黄醇、4-己基间苯二酚或这两种化合物的组合的测试组合物进行如上所述的CRAPBII表达测试。对于该系列测试，使用了表1中描述的水包油乳液基质。视黄醇和/或4-己基间苯二酚均包括在该油包水乳液基质中。CRAPBII表达的值在下表3中记录。对于组合，也记录了如上所述计算的％增效。
表3：CRAPBII表达
如表3中所示，CRAPBII表达与测试范围内的视黄醇浓度正相关并且对其相当敏感。以低的量使用的4-己基间苯二酚总体上改善了CRAPBII表达，而较高量的4-己基间苯二酚总体上减少了CRAPBII表达。在视黄醇的浓度不超过组合物的约0.075重量％时惊人地观察到增效响应。此外，当4-己基间苯二酚的浓度为约0.5重量％(大约50μg/ml)时该增效被大大地放大。
实例3：Bay 11-7082和视黄醇-CRAPBII的表达
对一系列含有视黄醇、Bay 11-7082或这两种化合物的组合的测试组合物进行CRAPBII表达测试。对于该系列测试，将下面表4中所示的水包油乳液基质用于补足测试组合物的余量。
表4：水包油乳液基质
| 成分 | 浓度(重量百分比) |
| 水 | 55.7123 |
| 丙烯酰二甲基牛磺酸铵/VP | 0.5 |
| 乙二胺四乙酸二钠 | 0.1 |
| 甘油 | 5.0 |
| 丁二醇 | 2.0 |
| PEG-8 | 5.0 |
| 鲸蜡醇：鲸蜡硬脂醇聚醚-20 | 3.0 |
| 硬脂醇；鲸蜡硬脂醇聚醚-20 | 3.0 |
| 甲氧基肉桂酸乙基己酯 | 2.0 |
| 异十六烷 | 1.5 |
| PPG-15硬脂基醚；BHT | 4.5 |
| 季戊四醇四(乙基己酸)酯 | 7.0 |
| 乳木果油(牛油树脂) | 1.0 |
| 生育酚乙酸酯 | 0.25 |
| BHT | 0.1 |
| 聚二甲基硅氧烷 | 2.0 |
| 环己硅氧烷；环戊硅氧烷 | 2.01 |
| 聚丙烯酰胺；C13-14异链烷烃；月桂醇聚醚-7 | 2.0 |
| 尼龙-12 | 3.0 |
| 抗坏血酸 | 0.05 |
| 柠檬酸 | 0.25 |
| 氢氧化钠 | 0.0277 |
对于该系列测试，视黄醇包含在测试组合物中。然而，Bay11-7082不包含在油包水乳液基质中，而是包含在培养基中。下表5中记录了CRAPBII表达。对于组合，也记录了％增效。
表5：CRAPBII表达
如表5中所示，对于视黄醇与NF-kB抑制剂Bay11-7082的组合，发现CRAPBII表达得到增效。虽然0.04％视黄醇与1μM的BAY11-7082的组合显示了增效的CRAPBII表达，但对于0.04％视黄醇与10μM的Bay11-7082的组合未观察到增效。
数据证明，在其中类视色素以不超过组合物的约0.075重量％的浓度存在的组合物中NF-KB抑制剂和类视色素的组合产生了视黄醇活性的令人惊奇的、增效的增加。
Claims (7)
1.一种组合物，所述组合物包含视黄醇、4-己基间苯二酚和美容上可接受的局部用载体，其中所述组合物中视黄醇的量为所述组合物的0.01重量％至0.075重量％，所述组合物包含0.1重量％至0.5重量％的4-己基间苯二酚。
2.根据权利要求1所述的组合物，所述组合物包含0.01重量％至0.06重量％的视黄醇。
3.一种处理哺乳动物皮肤的方法，所述方法包括将根据权利要求1所述的组合物施用至所述皮肤，所述方法不用于治疗目的。
4.根据权利要求3所述的方法，其中所述皮肤需要皮肤老化迹象方面的改善。
5.根据权利要求4所述的方法，其中所述皮肤需要紧实度、肌理、细纹或皱纹的出现或者弹性损失方面的改善。
6.根据权利要求3所述的方法，其中所述皮肤需要痤疮的迹象方面的改善。
7.根据权利要求6所述的方法，其中所述皮肤包括疵点、病损或丘疹、出现前丘疹、黑头和/或白头。
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| US8084504B2 (en) | 2009-10-02 | 2011-12-27 | Johnson & Johnson Consumer Companies, Inc. | High-clarity aqueous concentrates of 4-hexylresorcinol |
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