CN114073769A - 包含至少一种糖基化类黄酮的组合物及其在化妆品或皮肤病学中的用途 - Google Patents
包含至少一种糖基化类黄酮的组合物及其在化妆品或皮肤病学中的用途 Download PDFInfo
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- CN114073769A CN114073769A CN202110952547.1A CN202110952547A CN114073769A CN 114073769 A CN114073769 A CN 114073769A CN 202110952547 A CN202110952547 A CN 202110952547A CN 114073769 A CN114073769 A CN 114073769A
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- pyrene
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- myricitrin
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Abstract
本发明涉及包含至少一种糖基化类黄酮作为活性成分的化妆品和/或皮肤病学组合物,其特征在于,该组合物还包含作为活性成分的能够限制或阻止苯并[a]芘、环戊[c,d]芘、二苯并[a,h]蒽或二苯并[a,l]芘类型的多环芳烃渗透到角质形成细胞中的化合物和/或以其吸收蓝光的能力为特征的化合物。
Description
技术领域
本发明涉及包含至少一种糖基化类黄酮的组合物,其可用于局部应用,用于治疗引起IL31产生的非过敏性病理学状况(pathologie),特别是影响皮肤的病理学状况,并且选自瘙痒症、特应性皮炎 (dermatite atopique)、湿疹、银屑病、结节性痒疹。
背景技术
皮肤是保护人体免受环境影响的主要屏障。这种环境由外源性因素组成,这些因素会产生物理、化学甚至生物应力,从而增加皮肤老化和发生不同严重程度的皮肤病理学状况的风险。现在公认污染和太阳辐射对皮肤有协同和负面影响(The skin agingexposome,Krutmann J. et al.Dermatol Sci.2017Mar;85(3):152-161)。
污染可以呈现不同的形式:大气污染、水污染、全球变暖(热污染)、通过工业甚至日常用品的成分(油漆、溶剂、胶等)造成的化学污染。
由可见光、UVA/UVB还有蓝光组成的太阳辐射会导致产生IL-1、 IL-6、IL-8型炎性细胞因子。最近的其他研究表明,由太阳以及由计算机、平板电脑、电视、移动电话类型的设备的所有照明所发出的蓝光会通过增加角质形成细胞和黑素细胞内的自由基的比率来影响光老化。这也会导致色素斑的形成(Duteil et al,Differences in visible light-induced pigmentation according to wavelengths:a clinical and histologicalstudy in comparison with UVB exposure,Pigment Cell Melanoma Res,2014vol.27(5)pp.822-6T)。
大气污染在皮肤病理学状况中也起着重要作用,特别是多环芳烃(HAP),其来自木材的燃烧,但也来自废气(尤其在城市地区)。在最近的一项研究中,研究人员表明,在人类皮肤的外植体中,HAP 的代谢导致产生易于消除的代谢物,但也有一小部分与DNA反应而诱导突变。这项研究还表明,在太阳UV下的暴露会降低皮肤细胞代谢HAP的能力,从而可能在组织中累积(Metabolism and genotoxicity of polycyclic aromatic hydrocarbons inhuman skin explants:mixture effects and modulation by sunlight,Anne vonKoschembahr et al., Archives of Toxicology volume 94,pages 495–507(2020);Solar simulated light exposure alters metabolization and genotoxicity inducedby benzo[a]pyrene in human skin,Anne von Koschembahr et al., ScientificReports volume 8,Article number:14692(2018))。
代谢降低可导致苯并[a]芘(B[a]P)的累积以及长期毒性。
某些皮肤病理学状况不存在炎症成分,例如瘙痒症的情况即是如此,但其特征是分泌某些细胞因子如IL-31(白细胞介素31)(IL-31 is crucial for induction ofpruritus,but not inflammation,in contact hypersensitivity,Ayako Takamori etal.,Sci Rep(2018)8:6639)。
IL-31是IL-6家族的一种细胞因子,它不产生活化的CD4+T淋巴细胞,表明它参与了Th2型免疫应答的发展。已经证明,对于IL-31,转基因小鼠出现特应性皮炎类型的炎症性皮肤。此外,Il-31看来与瘙痒症有关。
有各种治疗瘙痒的方式,尤其是由IL-31产生增加引起的瘙痒。治疗性抗体anti-IL-31已被开发并用于治疗瘙痒症或特应性皮炎,例如Nemolizumab,这是一种由Galderma公司在中度至重度特应性皮炎的临床试验中开发的抗体。其他anti-IL-31抗体因其在这种应用领域中的用途也被描述(US10273297、EP2734549)。然而,通过治疗性抗体进行的治疗并非没有副作用,因为它们对皮肤的角质形成细胞所产生的IL-31不是特异性的。此外,这些治疗是昂贵的。因此确实需要一种解决方案能够通过局部水平的治疗来减弱角质形成细胞中 IL-31的产生。
发明内容
本发明人首先观察到,在与暴露于蓝光协同地暴露于HAP之后,通过角质形成细胞的IL-31的产生得到增加。
本发明人已经令人惊讶地证明,糖基化类黄酮类型的化合物使得能够减少由暴露于HAP和蓝光所诱导的通过角质形成细胞的IL-31 的产生。
要解决的技术问题
本发明的一个目的在于提供一种化妆品和/或皮肤病学组合物,其使得能够减少皮肤处的IL-31的产生并治疗或预防影响皮肤的病理学状况,所述病理学状况选自瘙痒症、特应性皮炎、湿疹、银屑病、结节性痒疹。
发明详述
本发明的一个目的在于提供包含至少一种糖基化类黄酮作为活性成分的化妆品和/或皮肤病学组合物,其特征在于,该组合物还包含作为活性成分的能够限制或阻止苯并[a]芘、环戊[c,d]芘、二苯并[a,h]蒽或二苯并[a,l]芘类型的多环芳烃渗透到角质形成细胞中的化合物和/或以其吸收蓝光的能力为特征的化合物。
术语“化妆品组合物”被理解为是指以下的任何物质或组合物:其旨在与人体的各个表面部分(表皮、毛发和毛细血管系统、指[趾] 甲、嘴唇和外生殖器)或与牙齿和口腔粘膜接触,专门地或主要地是为了清洁它们,给它们加香,改变它们的外观,保护它们,保持它们的良好状态或纠正体味。
术语“皮肤病学组合物”被理解为是指旨在治疗或预防皮肤疾病或用于常规皮肤护理的任何物质或组合物。
术语“活性成分”被理解为是指用在治疗性或化妆品组合物的配制剂中的提取或合成的天然或化学物质,特征是它对病理学状况或症状具有治疗或预防的有益效果。
术语“类黄酮”被理解为是指属于类黄酮家族的任何化合物,其特征为由三个碳:C6-C3-C6所连接的两个芳环而形成的相同基本结构,分成的类别为黄酮、黄酮醇、二氢黄酮醇(flavononols) 或二氢黄酮醇(dihydroflavonols)、黄烷酮、橙酮(aurones)、查耳酮和二氢查耳酮。
术语“糖基化类黄酮”被理解为是指携带至少一种选自葡萄糖、半乳糖、鼠李糖的糖的上述类别的任何化合物。
根据本发明的类黄酮是天然或合成的类黄酮。类黄酮天然存在于水果、蔬菜、茶、酒中,也存在于某些药用植物如山楂、银杏、西番莲、胡椒薄荷、朝鲜蓟、或者甚至楝树、Goyabier中,还有许多其他植物和花卉中(非穷举清单)。可以对它们进行提取。众多操作方案已经被公开并且可由本领域技术人员获取。
在本发明的一种实施方案中,根据本发明的组合物还包含作为活性成分的能够限制或阻止苯并[a]芘、环戊[c,d]芘、二苯并[a,h]蒽或二苯并[a,l]芘类型的多环芳烃渗透到角质形成细胞中的化合物。
表述“限制或阻止苯并[a]芘、环戊[c,d]芘、二苯并[a,h]蒽或二苯并[a,l]芘类型的多环芳烃渗透到角质形成细胞中的化合物”被理解为是指任何物质将HAP类型的污染剂挡在皮肤外表面上的能力。换句话说,限制或阻止HAP渗透到角质形成细胞中相当于将角质形成细胞维持在其生理平衡状态,也即当结构和功能没有改变时,它们不表现出任何炎症的特征或标记,例如细胞因子(如TNFα)的合成和释放(The influence of PM2.5 on lunginjury and cytokines in mice.Yang J,et al.,Exp Ther Med.2019Oct;18(4):2503-2511)。这种极具攻击性的白细胞介素会引起皮肤膜和细胞降解,导致皮肤的炎症和老化(Wang Y.,et al.,Mech Ageing Dev.2019Dec,184:111160)。
也被称作HAP的多环芳烃是以下类型的:苯并[a]芘(C20H12)、环戊[c,d]芘(C18H10)、二苯并[a,h]蒽(C22H14)或二苯并[a,l]芘(C24H18或C24H14)或这些化合物的衍生物。这些化合物通过化石材料的燃烧(特别是通过柴油发动机)以气体或颗粒形式产生。因此HAP尤其存在于城市空气污染中。
烃在人体器官内的经过通过吸入,通过摄入,也通过经由皮肤的传递来进行。这是因为,本发明人在此发明的先前工作中已经表明,人的角质形成细胞暴露于HAP增加了角质形成细胞中IL-31的产生。
在本发明的一种实施方案中,根据本发明的组合物还可包含以其吸收蓝光的能力为特征的化合物。
表述“吸收蓝光的能力”被理解为是指任何物质通过限制皮肤对所述蓝光的暴露来防止蓝光对皮肤老化和皮肤损伤的负面影响的能力。
波长介于400至500纳米(nm)之间的蓝光是介于400至800纳米之间的可见光谱的一部分。它由太阳辐射发射,但也由所有带有基于电致发光二极管(也被称为LED)的屏幕的电子设备发射,所述电子设备例如是智能手机、平板电脑、电视或计算机。
由这种蓝光发出的能量对于太阳辐射来说是大约40J/cm2,并且对于电子设备的屏幕来说是大约10J/cm2。
在一种优选的实施方案中,该糖基化类黄酮是糖基化黄酮醇。黄酮醇构成了一个类别的类黄酮,其特征是在类黄酮的基本骨架的中心杂环上的C3处的酚官能团和C4处的羰基官能团C=O。糖基化黄酮醇可以是单糖苷、二糖苷或三糖苷的形式。在单糖苷中有黄芪苷(R3 处含有葡萄糖的莰非醇)、杜鹃黄苷(R3处含有鼠李糖的杜鹃黄素)、金丝桃苷(R3处含有半乳糖的槲皮素)、异槲皮素(R3处含有葡萄糖的槲皮素)、杨梅苷(R3处含有鼠李糖的杨梅树皮素)、栎皮苷(R3 处含有鼠李糖的槲皮素)、芸香苷(R3处含有芸香糖的槲皮素)、黄鼠李苷(R3处含有三纯苷(triholoside)的鼠李素)、黄柏苷(R7 处含有葡萄糖和R8处含有叔戊醇的莰非醇)、绣线菊苷(R4’处含有葡萄糖的槲皮素)和曲克芦丁(R3处含有芸香糖并含有3羟乙基的槲皮素)。
在二糖苷中有山柰苷(kaempferitrine)(R3处含有鼠李糖和R7 处含有鼠李糖的莰非醇)、刺槐苷(R3处含有刺槐糖和R7处含有鼠李糖的莰非醇)、淫羊藿苷(含有R3处的鼠李糖和R4处的葡萄糖以及异戊二烯基和甲氧基的莰非醇)。
在一种优选的实施方案中,糖基化类黄酮是鼠李糖基化的或葡萄糖基化的,优选鼠李糖基化的。
在一种优选的实施方案中,鼠李糖基化类黄酮选自杨梅苷(5,7- 二羟基-3-[(2S,3R,4R,5R,6S)-3,4,5-三羟基-6-甲基烷-2-基]氧基 -2-(3,4,5-三羟基苯基)色烯-4-酮)、栎皮苷(2-(3,4-二羟基苯基)-5,7- 二羟基-3-[[(2S,3R,4R,5R,6S)-3,4,5-三羟基-6-甲基-2-四氢吡喃基]氧基]-4-色烯酮)、山柰苷(C27H30O14)、杜鹃黄苷(2-(3,4-二羟基苯基)-7- 羟基-5-甲氧基-3-[(2S,3R,4R,5R,6S)-3,4,5-三羟基-6-甲基烷-2-基]氧基色烯-4-酮)、淫羊藿苷(5-羟基-2-(4-甲氧基苯基)-8-(3-甲基丁-2- 烯基)-7-[(2S,3R,4S,5S,6R)-3,4,5-三羟基-6-(羟甲基)烷-2-基]氧基 -3-[(2S,3R,4R,5R,6S)-3,4,5-三羟基-6-甲基烷-2-基]氧基色烯-4-酮)或这些化合物中至少两种的混合物。该混合物可由以下物质构成:杨梅苷和栎皮苷,或杨梅苷和山柰苷或杨梅苷和杜鹃黄苷,或杨梅苷和淫羊藿苷,或栎皮苷和杜鹃黄苷,或栎皮苷和淫羊藿苷,或杜鹃黄苷和淫羊藿苷。该混合物还可由三种鼠李糖基化类黄酮构成,例如杨梅苷、栎皮苷和杜鹃黄苷,或杨梅苷、栎皮苷和淫羊藿苷,或杨梅苷、山柰苷和杜鹃黄苷,或杨梅苷、杜鹃黄苷和淫羊藿苷,或例如栎皮苷、杜鹃黄苷和淫羊藿苷。该清单并不是限制性的。
在一种优选的实施方案中,该鼠李糖基化类黄酮是杨梅苷。
在本发明的一种实施方案中,所述能够限制或阻止苯并[a]芘、环戊[c,d]芘、二苯并[a,h]蒽或二苯并[a,l]芘类型的多环芳烃渗透到角质形成细胞中的化合物选自多糖、环状糊精类型的环状多糖、乳化剂和亲脂化透明质酸。
皮肤由不同的层和不同的细胞类型构成。最外层即表皮由角质形成细胞、黑素细胞和朗格汉斯细胞构成。这种表皮分为四层,以组成它们的角质形成细胞的成熟程度为特征。从最内到最外层依次为:基底层、Malpighi层、颗粒层和角质层。这个角质层是一种疏水屏障,它确保保护皮肤所需的屏障功能。
考虑到角质层的疏水特性,亲脂性物质如多环芳烃可容易地渗透到角质形成细胞中。
多糖也被称为聚糖、多糖(polyosides)、多糖(polyholosides) 或复合糖类,其是由多个单糖通过糖苷键彼此连接而构成的糖类家族的聚合物。
多糖具有亲水性能。因此,并且由于HAP的疏水性,这些多糖形成了阻止这些物质渗透的亲水性物理屏障。
本发明范围内使用的多糖可以是环状糊精类型的环状多糖,其包括阿尔法-环状糊精(α-环状糊精,C36H60O30)、贝塔-环状糊精(β- 环状糊精,C42H70O35)、伽玛-环状糊精(γ-环状糊精,C48H80O40)。环状糊精已知形成分子笼,允许包封各种分子。它们被广泛地用于农产食品加工(agroalimentaire)和药物领域中。它们的天然来源是一种巨大的优势。在本发明的范围内,环状糊精类型的环状多糖通过其在葡萄糖聚合物(6-9个残基)的环状形式的中心的“疏水笼”类型的分子结构来参与HAP的捕获。
内部疏水且外部亲水的环状糊精捕获HAP类型的分子并生成所述HAP的分子陷阱。
因此,本发明的目的之一在于一种组合物,该组合物包含作为活性成分的至少一种糖基化类黄酮,以及环状糊精,该糖基化类黄酮更特别地是杨梅苷(myricitrine)、栎皮苷(quercitrine)、杜鹃黄苷 (azaleine)、淫羊藿苷(icariine)或这些化合物中至少两种的混合物。
在本发明的范围内,能够限制或阻止多环芳烃渗透到角质形成细胞中的化合物是乳化剂。乳化剂是能够产生乳液的化合物。任何乳化剂均具有亲水端和疏水端,使得亲水和疏水物质的混合物成为可能。多环芳烃本质上是亲脂性和疏水性的。
在本发明的组合物中加入环状糊精类型的化合物旨在在HAP或HAP混合物与亲水性化合物之间形成乳液,以限制或阻止所述HAP 渗透到角质形成细胞中。
可在本发明组合物的范围内使用的乳化剂是:卵磷脂、磷脂、羊毛脂或蜂蜡(天然乳化剂)、硬脂酸甘油酯、聚山梨酯60和以名称销售的PEG-6/PEG32/硬脂酸乙二醇酯的混合物、槐糖脂类型的糖脂或皂苷的辛基葡糖苷、脂蛋白(非穷尽且非限制性的清单)。
因此,本发明的目的之一在于一种组合物,该组合物包含作为活性成分的至少一种糖基化类黄酮,以及乳化剂,该糖基化类黄酮更特别地是杨梅苷、栎皮苷、杜鹃黄苷、淫羊藿苷或这些化合物中至少两种的混合物。
在本发明的范围内,能够限制或阻止多环芳烃渗透到角质形成细胞中的化合物还是亲脂化透明质酸。
亲脂化透明质酸作为乳化基质具有亲脂性和亲水性的两亲性。这种分子在构成亲脂化部分的脂肪链处捕获亲脂性分子如B[a]P。
因此,本发明的目的之一在于一种组合物,该组合物包含作为活性成分的至少一种糖基化类黄酮,以及亲脂化透明质酸,该糖基化类黄酮更特别地是杨梅苷、栎皮苷、杜鹃黄苷、淫羊藿苷或这些化合物中至少两种的混合物。
类胡萝卜素将胡萝卜素和叶黄素家族的分子聚集在一起。主要的类胡萝卜素是虾青素、番茄红素、β-胡萝卜素、叶黄素和玉米黄质。类胡萝卜素是脂溶性的,具有公认的抗氧化和光保护性能。因此,类胡萝卜素因其能够吸收蓝光的能力而可被用在根据本发明的组合物中 (Blue-Violet Light Irradiation Dose Dependently Decreases Carotenoidsin Human Skin,Which Indicates the Generation of Free Radicals.Vandersee S.etal.,Oxidative Med Cell Longev.Volume 2015, Article ID 579675)。
因此,本发明的目的之一在于一种组合物,该组合物包含作为活性成分的至少一种糖基化类黄酮,以及环状糊精或乳化剂或亲脂化透明质酸,以及类胡萝卜素,该糖基化类黄酮更特别地是杨梅苷、栎皮苷、杜鹃黄苷、淫羊藿苷或这些化合物中至少两种的混合物。
类黑精是在Maillard反应期间通过糖和氨基酸之间的组合形成的高分子量的棕色异质聚合物。由于它们的结构和发色性能(吸收波长为400-500nm),它们是良好的蓝光过滤器(Coffee melanoidins: structures,mechanisms of formation and potentialhealth impacts, Moreira AS,et al.,Food Funct.2012Sep;3(9):903-15)。
因此,本发明的目的之一在于一种组合物,该组合物包含作为活性成分的至少一种糖基化类黄酮,以及环状糊精或乳化剂或亲脂化透明质酸,以及类黑精家族的化合物,该糖基化类黄酮更特别地是杨梅苷、栎皮苷、杜鹃黄苷、淫羊藿苷或这些化合物中至少两种的混合物。
在本发明的一种实施方案中,本发明的组合物被配制用于局部应用。
可用于本发明的组合物可呈任何常规用于局部应用的盖仑制剂的形式,特别是以下的形式:水性、醇性或水醇性溶液或悬浮液,或油性悬浮液或洗剂或乳浆类型的溶液或分散体,乳型液体或半液体稠度的乳液(通过将脂肪相分散在水相(H/E)或反之(E/H)中而获得),或含水或无水凝胶,软膏,原样使用或掺入赋形剂中的松散或压实的粉末,或微胶囊或微颗粒的形式,或离子和/或非离子类型的囊泡分散体。
已知地,根据本发明的组合物还可包含化妆品、药物或皮肤病学领域中常用的佐剂,例如亲水性或亲脂性胶凝剂、亲水性或亲脂性活性剂、防腐剂、抗氧化剂、溶剂、香料、填料、过滤剂、杀菌剂、气味吸收剂和着色材料。本领域技术人员知道如何根据所需盖仑剂型选择佐剂及其用量。
在本发明的一种优选实施方案中,根据本发明的组合物被配制成雾(brume)的形式,如实施例8中所述。
在一种实施方案中,本发明涉及糖基化类黄酮化合物,优选鼠李糖基化黄酮醇,其用于其在治疗引起IL31产生的非过敏性病理学状况中使用,该病理学状况特别是影响皮肤的病理学状况,并且选自瘙痒症、特应性皮炎、湿疹、银屑病、结节性痒疹。
术语“治疗”被理解为是指病理学状况的至少一种症状的减弱或消失,可以是角质形成细胞产生的IL-31减少、对抓挠的敏感性降低、皮肤炎症减少以及直到一种或多种症状消失。
本发明人已经表明,人角质形成细胞对HAP和/或蓝光的暴露导致所述角质形成细胞产生的IL-31增加。此外,作者已经表明,瘙痒症或特应性皮炎中的皮肤瘙痒与IL-31的这种产生有关(New mechanism underlying IL-31-induced atopic dermatitis,Meng J.elal.,J Allergy Clin Immunol.2018May;141(5):1677-1689;IL-31:A new key player indermatology and beyond,IS et al.,J Allergy Clin Immunol.2018Mar;141(3):858-866)。本发明的目的之一涉及糖基化类黄酮化合物,优选鼠李糖基化黄酮醇,用于其在治疗引起IL-31产生的非过敏性病理学状况中的用途,该病理学状况特别是影响皮肤的病理学状况,并且选自瘙痒症、特应性皮炎、湿疹、银屑病、结节性痒疹。
瘙痒症在临床上被定义为想要抓挠的主观和不愉快的感觉。有一种始于皮肤的感觉成分,其通过背部神经结传输并在中枢神经系统中进行处理。通常,运动反应以想要抓挠的形式出现。抓挠会损伤皮肤并引发炎症反应,瘙痒会进一步加剧炎症反应。
特应性皮炎也被称为特应性湿疹,它是一种皮肤慢性炎症性疾病。它优先在婴儿和儿童中发展,但可持续存在,有时甚至在青少年和成人中出现。它的特征是与湿疹样病变(发红和瘙痒、水泡、渗液和结痂)相关的皮肤干燥(干燥症),这些病变在突然发作时出现。
湿疹是一种综合征,是多种疾病共有的一组临床体征和症状,可能对应于多种炎症性皮肤病。这些疾病的特征是瘙痒(瘙痒症)、皮肤充血性发红(红斑)和皮疹。
银屑病是一种慢性炎症性疾病,其特征是斑块边界清楚、红色、隆起,并覆盖有白色鳞屑或容易脱落的银色薄膜。
痒疹或结节性痒疹是一种皮肤病(皮肤疾病),其特征是发痒病变(抓挠),有些搔破和/或伴有搔痕,伴有剧烈瘙痒,可影响睡眠。抓挠可维持变成结痂的病变。可分为最常继发于昆虫叮咬的急性痒疹 (持续时间少于6周),以及可能是皮肤病学、神经学、精神病学、系统性来源的慢性痒疹。
在本发明的一种优选实施方案中,用于治疗引起IL31产生的非过敏性病理学状况(该病理学状况特别是影响皮肤的病理学状况,并且选自瘙痒症、特应性皮炎、湿疹、银屑病、结节性痒疹)的鼠李糖基化黄酮醇选自杨梅苷、栎皮苷、山柰苷、杜鹃黄苷、淫羊藿苷或这些化合物中至少两种的混合物。该混合物可由以下物质构成:杨梅苷和栎皮苷,或者杨梅苷和杜鹃黄苷,或者杨梅苷和淫羊藿苷,或者栎皮苷和杜鹃黄苷,或者栎皮苷和淫羊藿苷,或者杜鹃黄苷和淫羊藿苷。该混合物还可以由三种鼠李糖基化类黄酮构成,例如杨梅苷、栎皮苷和杜鹃黄苷,或杨梅苷、栎皮苷和淫羊藿苷,杨梅苷、杜鹃黄苷和淫羊藿苷或栎皮苷、杜鹃黄苷和淫羊藿苷。
附图说明
-图1显示细胞活力测试,测量在不同剂量杨梅苷的存在下人角质形成细胞的活细胞的百分比;
-图2显示单独地或与蓝光联合的α苯并芘对通过人角质形成细胞的IL-31的产生的影响。对蓝光的暴露以焦耳/平方厘米(J/cm2)表示。对α苯并芘的暴露以微摩尔(μM)表示;
-图3显示杨梅苷对预先暴露于α苯并芘和蓝光的角质形成细胞对IL-31释放(以pg/ml计)的影响;
-图4表示通过细胞总蛋白的量标准化的IL31的浓度,说明在角质形成细胞暴露于蓝光和苯并-a-芘之后,与单独的杨梅苷的效果相比的单独的虾青素的效果;
-图5表示对于两种浓度的杨梅苷和虾青素来说,IL31释出的抑制值。
具体实施方案
实施例
实施例1:人角质形成细胞的培养
此研究中使用的细胞是从一名30岁的捐献者皮肤手术后提取的正常人角质形成细胞(KHN)的原代培养物。
它们在以下的KSFM完全培养基中培养:角质形成细胞-SFM (17005-31,Gibco),含有L-谷氨酰胺,补充以人重组表皮生长因子(EGFhr,10450-013,Gibco),牛垂体提取物(BPE,13028-014, Gibco)和1%青霉素链霉素(15070-063,Gibco),在37℃下和5%CO2的气氛中,直至80%的汇合。
实施例2:角质形成细胞的活力测试
在第0天,所述正常人角质形成细胞在培养基中在96孔板中温育。如实施例1所述,使所述细胞在37℃下在加有5%CO2的气氛中稳定 24小时。
在第1天,细胞用以不同浓度溶解在培养基中的杨梅苷进行处理。
培养物在37℃和5%CO2下保持24小时。
KHN的活力使用细胞增殖试剂盒II(XTT)(11465015001,Roche) 根据制造商的指示进行评价。
XTT系统(一种比色法)是一种用于量化线粒体活性的测试。这种简单、精确且结果可重复的方法可被用作活力测试。这种测试基于通过细胞线粒体呼吸链中存在的“琥珀酸-四氮唑还原酶”系统将黄色四氮唑盐XTT裂解为橙色甲(formazan)。因此,转化仅发生在代谢活跃的细胞(这表明是活细胞)中。
处理的细胞的活力以未处理的对照的百分比来表示:与未处理参照(témoin)相比,将细胞活力降至低于线粒体活性的80%限值的处理被认为对细胞具有细胞毒性。相反,数据的增加是线粒体活性的标志,并且甚至可是细胞增殖的标志。
根据实施例1培养的角质形成细胞被暴露于0.21-50μM的杨梅苷范围内。参照对照(Témoin)代表不含佐剂的培养基。对照物+10%DMSO代表有毒培养基,作为细胞毒性的参照。实验表明,杨梅苷对角质形成细胞没有任何毒性作用。
实施例3:在培养的角质形成细胞上使用鼠李糖基化黄酮醇类型的类黄酮
根据实施例1培养的角质形成细胞暴露于20μM的α苯并芘 (B[a]P),或暴露于40J/cm2的蓝光(450nm),或暴露于相同剂量的二者的组合。用10μM-50μM范围内的量的杨梅苷的处理被应用于预前接受B[a]P和蓝光组合的细胞。
在处理24小时后,通过免疫测定试剂盒(ELISA)进行由细胞释出的白细胞介素31(IL-31)的测定。使用人IL-31ELISA MAXTM Deluxe Set(445704,Biolegend)进行IL-31的测定。
α苯并芘和蓝光的组合导致通过角质形成细胞的IL-31增加 +111%。实验表明,从20μM开始加入杨梅苷会导致用20μM的B[a]P 和40J/cm2蓝光预处理的细胞中IL-31的产生减少-100%。
实施例4:对患者使用鼠李糖基化黄酮醇类型的类黄酮
针对脸上有皮肤不适问题(瘙痒、瘙痒症等)的在城市环境中工作的一组志愿者[20至60岁的白种人和亚洲人类型的女性],将实施例 5的霜施用到一半脸上并且将不含杨梅苷的安慰剂霜施用到另一部分的脸上。
从施用开始并且在早晚使用的八天期间,志愿者们通过完成问卷调查来给予她们的感官感受-产品的新鲜度、柔软性和易于施用性,以及瘙痒停止、镇静类型的生理感受。在测试结束时进行统计分析,以证明含有杨梅苷的产品的有效性。
实施例5:用于城市或易发痒皮肤的镇静和软化霜。
A-水相
甘油2.0%,己二醇3.0%,黄原胶0.5%,杨梅苷0.5%,防腐剂 qs(适量),卡波姆0.35%,NaOH 0.35%,水适量至(qsp.)100%
B-油脂相
角鲨烷15%,鲸蜡醇2%,花生醇/山萮醇/花生醇葡糖苷1%,硬脂酸甘油酯5%,防腐剂和香料适量
实施例6:旨在用于具有特应性倾向的城市皮肤的舒缓保湿霜
A–油脂相
Ceteareth-2 3.5%,Ceteareth-21 2-4%,小麦胚芽油3%,环甲硅油(Cyclomethicone)7%,棕榈酸辛酯8%
B-水相
甘油7.0%、己二醇3.0%、防腐剂适量、水适量至100%
C-在低于40℃的温度下在乳液中添加的成分
透明质酸钠0.1%,杨梅苷1.0%,水5%,生育酚0.05%,维生素A棕榈酸酯0.1%,磷脂0.5%,神经酰胺3 0.1%,聚丙烯酰胺 &C14-13异链烷烃&Laureth-7 2-3.5%
实施例7:旨在用于暴露于蓝光或具有色素斑的皮肤的霜和乳。
A–油脂相
甘油单硬脂酸酯2%,PEG-100硬脂酸酯3%,C12-C15烷基苯甲酸酯10%,二甲硅油(Dimethicone)5%,生育酚乙酸酯1%,辛基三嗪酮(Uvinul T150)1.5%,丁基甲氧基二苯甲酰甲烷(Eusolex 9020)2.0%,鲸蜡硬脂醇1%
B-水相
水适量至100%,栎皮苷0.5%,防腐剂0.6%,甘油7%,己二醇3.0%,卡波姆0.5%,EDTA四钠0.2%,透明质酸钠0.1%,NaOH 0.5%,防腐剂+香料适量。
C-在低于40℃的温度下在乳液中添加的成分
生育酚乙酸酯0.1-1%、吡哆醇0.01-0.05%、维生素A棕榈酸酯 0.01-1%、d-泛醇0.1-1%、柠檬酸0.1-0.5%、葡萄糖酸锌0.1-1%,柠檬酸三钠1-2.5%,水5%。
实施例8:旨在用于暴露于电子设备的蓝光下的皮肤的防护雾 (brume)
水适量至100%,山柰苷2%,防腐剂0.6%,甘油7%,己二醇 3.0%,卡波姆0.5%,EDTA四钠0.2%,透明质酸钠0.1%,NaOH 0.5%,防腐剂+香料适量。
实施例9:虾青素和杨梅苷对暴露于蓝光和苯并-a-芘(BaP)的组合作用的原代角质形成细胞释出IL31的作用的对比。
虾青素(3,3’-二羟基-4,4’-二氧代β-胡萝卜素)是一种类胡萝卜素,特别是由微藻合成,这种分子吸收400nm-500nm的波长。
苯并-a-芘(CAS 50-32-8)是毒性最强的HAP(多环芳烃)之一。
细胞的处理按照下述方式进行。
a)细胞类型:来自人表皮的原代角质形成细胞(捐献者39岁,标号:KER110,批号KER110049,Biopredic)
b)培养条件:96孔板,37℃,5%CO2,接种20,000个细胞/cm2
c)培养基:KSFM(Gibco ref:10144892)+1%青霉素/链霉素
d)利用化合物的处理:接种后24小时进行,
e)利用化合物的温育时间:接触24小时
f)测试化合物的浓度:20μM和50μM的杨梅苷和虾青素
g)应激条件:在用化合物处理后进行;暴露于辐射(蓝光;40J/cm2; 450nm);然后在培养基中用20μM苯并-a-芘(CRM40071, Sigma-Aldrich)处理24小时。
根据以下程序进行分析。
-对照批次:除培养基更新外不进行任何处理
-取样:
培养基
细胞颗粒状沉淀
-终点/读出:
从培养基测定IL-31(人IL-31ELISA MAX Deluxe; Biolegends)
从细胞残渣测定总蛋白质(Bradford法)
结果:IL31的剂量
在图4中,结果表示为通过总细胞蛋白质的量标准化的IL-31浓度。
在图5中,根据以下公式获得实验组的释出抑制值(%):
IL31释出抑制%:%批次X=(IL31应激-IL31批次X)/(IL31 应激-IL31对照)X100
作为参考,参照组被视为最大功效(100%),而应激组(蓝光+BaP) 被视为最小功效(0%)。
Claims (10)
1.包含至少一种糖基化类黄酮作为活性成分的化妆品和/或皮肤病学组合物,其特征在于,该组合物还包含作为活性成分的能够限制或阻止苯并[a]芘、环戊[c,d]芘、二苯并[a,h]蒽或二苯并[a,l]芘类型的多环芳烃渗透到角质形成细胞中的化合物和/或以其吸收蓝光的能力为特征的化合物。
2.根据权利要求1的化妆品和/或皮肤病学组合物,其特征在于所述糖基化类黄酮是糖基化黄酮醇。
3.根据权利要求1或2的化妆品和/或皮肤病学组合物,其特征在于所述糖基化类黄酮是鼠李糖基化的或葡萄糖基化的,优选鼠李糖基化的。
4.根据权利要求3的化妆品和/或皮肤病学组合物,其特征在于所述鼠李糖基化类黄酮选自杨梅苷、栎皮苷、山柰苷、杜鹃黄苷、淫羊藿苷或这些化合物中至少两种的混合物。
5.根据权利要求4的化妆品和/或皮肤病学组合物,其特征在于所述鼠李糖基化类黄酮是杨梅苷。
6.根据前述权利要求中任一项的化妆品和/或皮肤病学组合物,其特征在于所述能够限制或阻止苯并[a]芘、环戊[c,d]芘、二苯并[a,h]蒽或二苯并[a,l]芘类型的多环芳烃渗透到角质形成细胞中的化合物选自多糖、环状糊精类型的环状多糖、乳化剂和亲脂化透明质酸。
7.根据前述权利要求中任一项的化妆品和/或皮肤病学组合物,其特征在于所述能够吸收蓝光的化合物选自类胡萝卜素或类黑精。
8.根据前述权利要求中任一项的化妆品和/或皮肤病学组合物,其特征在于其被配制用于局部应用。
9.糖基化类黄酮化合物,优选鼠李糖基化黄酮醇,其用于其在治疗引起IL31产生的非过敏性病理学状况中使用,该病理学状况特别是影响皮肤的病理学状况,并且选自瘙痒症、特应性皮炎、湿疹、银屑病、结节性痒疹。
10.根据权利要求9的糖基化类黄酮化合物,优选鼠李糖基化黄酮醇,选自杨梅苷、栎皮苷、山柰苷、杜鹃黄苷、淫羊藿苷或这些化合物中至少两种的混合物。
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