CN109700676A - 包含壬醛作为有效成分的防止脱发或促进生发用组合物 - Google Patents
包含壬醛作为有效成分的防止脱发或促进生发用组合物 Download PDFInfo
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- CN109700676A CN109700676A CN201811248475.7A CN201811248475A CN109700676A CN 109700676 A CN109700676 A CN 109700676A CN 201811248475 A CN201811248475 A CN 201811248475A CN 109700676 A CN109700676 A CN 109700676A
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- hair
- aldehyde
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- scalp
- tonic
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Abstract
本发明涉及包含壬醛或其药学上可接受的盐作为有效成分的防止脱发或促进生发用组合物，更具体地，本发明涉及包含壬醛或其盐作为有效成分的预防脱发、治疗脱发、促进生发或促进育发用药物组合物、准药物组合物、化妆品组合物及保健食品组合物。本发明的组合物不仅包含即使向作为慢性疾病的脱发长期给药也没有副作用的天然化合物，还对生发及育发显示出优秀且稳定的效果，因此可用作预防或治疗脱发剂、或者促进生发或育发用药品、准药物、化妆品、保健食品组合物。
Description
技术领域
本发明涉及包含壬醛或其药学上可接受的盐作为有效成分的防止脱发或促进生发用组合物，更具体地，本发明涉及包含壬醛或其盐作为有效成分的预防脱发、治疗脱发、促进生发或促进育发用药物组合物、准药物组合物、化妆品组合物及保健食品组合物。
背景技术
根据韩国国民健康保险工团健康保险政策研究院对2001年至2008年健康保险诊疗费支付资料的分析内容，2001年“脱发疾病”的实际诊疗患者数为10万3千人、2005年14万2千人、2008年16万5千人，过去七年增长了60％。按年龄段划分，20多岁至40多岁的实际诊疗患者数为11万4千人，占患者的69.5％，10岁以下患者数也超过2万2千人。以2008年为基准的性别实际诊疗患者数中，男性为8万4千人，女性为8万人，实际情况是男性略多于女性，以2008年为基准，“脱发”疾病的按照伤病分类的韩国国内健康保险实际诊疗患者数为斑型脱发症(13万人)、疤痕性脱发症(2万人)、雄激素性脱发症(9千人)、其他非疤痕性毛发损失(8千人)的顺序。
另一方面，根据2003年6月国际毛发及整形美容研讨会资料，全球约有2.5亿人患脱发，24～50岁中脱发发病率为30～65％左右。以2008年为基准，中国的脱发人口约为3亿，30多岁男性人口的30％以及50多岁男性人口的50％左右呈现出脱发症状，并且脱发患者数逐年增长10～15％左右。根据2007年度实施假发销售及毛发移植手术的企业对日本人的调查资料，日本人的脱发发病率为26.5％，估计脱发患者人数为1293万人左右。
目前，用于治疗脱发的制剂大可分为医药品和准药物以及化妆品。需要医生的处方才可以购买的专业药物有美国默克(Merck)公司研发销售的非那雄胺生发剂(Propecia)。作为非那雄胺生发剂的主成分的非那雄胺(finasteride)于1997年12月获得美国食品及药物管理局(FDA)的脱发治疗剂认可。非那雄胺作为抑制将睾酮转换为双氢睾酮(DHT，dihydrotestosterone)的5-α-还原酶的药剂，可以进行使软毛生长为粗且长的毛发的作用。这在短期内有改善脱发的效果，但是还伴有勃起功能障碍、性功能减退、男性乳房增大等的副作用。
作为因其安全性和有效性得到认可而不需要医生的处方也可以购买的普通医药品有米诺地尔(Minoxidil)，于1997年12月获得美国食品及药物管理局的首个涂敷型脱发治疗剂的认可。上述药品可以改善血液循环，开放钾隧道，从而存在可以促进毛发生长的效果，然而可以引起瘙痒、皮疹等的局部反应，还可以出现心动过速等现象。
代表性的作为从韩国食药厅获得防止脱发和育毛(肉毛)功能的许可的产品且借助韩国保健福祉部长官的告示而可以在超市或便利店销售的准药物有希杰狮王(CJLION)的“毛发力能力”、Moracle的“头发滋养剂”、LG生活健康的“Mo&Mor”等产品，化妆品类有洗发剂类或为维持或增进皮肤、毛发的健康而在头皮或毛发上使用的产品。
人的脱发周期大可分为生长期(anagen)、退行期(catagen)、休止期(telogen)。生长期为由于毛乳头的活动积极，细胞分裂旺盛，因而毛发迅速生长的时期。生长期的寿命因毛的种类而互不相同，在头发的情况下，为3～6年左右。生长期的毛发占整体毛发的80～90％，已开始进行脱发的人具有生长期变短、休止期长的毛发周期，在整体毛发中生长期毛发的比重减少。退行期为已结束毛发的生长期，毛发的生成减缓，最终停止细胞分裂及生长的时期，维持1～1.5个月左右，整体毛发的1％属于该阶段。休止期作为生长的最后阶段，为如下的阶段，即，由于毛囊和毛乳头完全分离，因而毛囊萎缩，毛根进一步上升并头发掉落。休止期持续3～4个月，整个毛发的4～14％属于该阶段。当结束休止期后毛乳头的活动再变得活跃时，造出新毛发的毛乳头，处于休止期的毛发被推出并完全脱离头皮。
另一方面，壬醛(Nonanal)作为多种植物的精油(essential oil)成分，作为可食用的安全物质已被知晓。作为芳香剂(flavor and fragrance agents)成分已获得美国香料与萃取物制造者协会(FEMA，Flavor and Extract Manufacturers’Association)、美国食品药品监督管理局(FDA，Food and Drug Administration)、韩国食品药品管理局(KFDA，Korea Food and Drug Administration)、欧洲委员会(COE，Council of Europe)以及食品添加剂专家委员会(JECFA，Joint FAO/WHO Expert Committee on Food Additives)的认可，并已在工业上用于呈现味道和香气的目的。并且，壬醛被用作如下的目的，当制造香水时，为了制造出天竺葵香、玫瑰香、茶香而使用，在食品中，为了制造出蜡味(waxy)、醛味、玫瑰味(rose)、鸢尾花味(fresh orris)、橘皮味(orange peel)、黄瓜味(cucumber)、柑橘味(citrus)、哈密瓜味(melon)、土豆味(potato)、椰子味(coconut)等而使用，并且为了给柑橘味增加香甜味而被作为芳香剂来使用。然而，还未表明对于壬醛的防止脱发或促进生发效果。
发明内容
为此，本发明者们试图发现可防止脱发或促进生发或育发的来源于天然的化合物。其结果，确认了壬醛可有效增进生发及育发且防止脱发，从而完成了本发明。
因此，本发明是为了解决如上所述的问题而提出的，本发明的目的在于，提供包含壬醛或其药学上可接受的盐作为有效成分的防止或治疗脱发用、或促进生发或育发用药物组合物。
本发明的再一目的在于，提供包含壬醛或其药学上可接受的盐作为有效成分的防止或改善脱发用、或者促进生发或育发用准药物组合物。
本发明的另一目的在于，提供包含壬醛或其化妆品学上可接受的盐作为有效成分的防止或改善脱发用、或者促进生发或育发用化妆品组合物。
本发明的还有一目的在于，提供包含壬醛或其食品学上可接受的盐作为有效成分的防止或改善脱发用、或者促进生发或育发用保健食品组合物。
然而，本发明要解决的问题不限于如上所述的问题，本领域技术人员可从以下描述中清楚地理解未提及的其他问题。
因此，本发明提供包含壬醛或其药学上可接受的盐作为有效成分的防止或治疗脱发用、或者促进生发或育发用药物组合物。
根据上述本发明的一优选实施例，上述组合物可用于增加选自由β-连环蛋白(β-catenin)、T细胞因子(T-cell factor)、淋巴样增强因子(lymphoid enhancer factor)及细胞周期蛋白D1(cyclin D1)组成的组的蛋白质的表达。
根据上述本发明的再一优选实施例，作为皮肤外用剂，上述药物组合物可以为选自由乳膏、凝胶、贴剂、喷雾剂、软膏剂、硬膏剂、洗剂、搽剂、糊剂及泥敷剂组成的组中的剂型。
并且，本发明提供包含壬醛或其药学上可接受的盐作为有效成分的防止或改善脱发用、或者促进生发或育发用准药物组合物。
并且，本发明提供包含壬醛或其化妆品学上可接受的盐作为有效成分的防止或改善脱发用、或者促进生发或育发用化妆品组合物。
根据上述本发明的另一优选实施例，上述化妆品组合物可以为头皮滋补剂、头皮洗剂、头皮乳膏、头皮乳清、头皮精华液、头皮浓缩精华、头皮护理、头皮护理剂、头皮洗发剂、头皮膜、头发滋养剂、头发洗剂、头发乳膏、喷发剂、发用摩丝、发胶、护发素、洗发水、润发液、发膜、焗油膏、眉毛生长剂、睫毛生长剂、睫毛营养剂、宠物用洗发水或宠物用润发液的剂型。
并且，本发明提供包含壬醛或其药学上可接受的盐作为有效成分的防止脱发或促进生发用保健食品组合物。
本发明是为了解决如上所述的问题而提出的，用于提供包含壬醛或其药学上可接受的盐作为有效成分的防止或改善脱发用、或者促进生发或育发用组合物。
本发明的组合物不仅包含即使向作为慢性疾病的脱发长期给药也没有副作用的天然化合物，还对生发及育发显示出优秀且稳定的效果，因此可用作预防或治疗脱发剂、或者促进生发或育发用药品、准药物、化妆品、保健食品组合物。
附图说明
图1为示出根据脱毛小鼠模型的各试样的给药日数的毛发的再生长照片。
图2为示出脱毛小鼠模型在涂敷各试样的期间所生长的毛的长度的图表。各值是5只小鼠的平均±标准误差。ns＝p＞0.05，**＝p＜0.01，***＝p＜0.001(单因素方差分析(one-way ANOVA)，Tukey’s试验)
图3示出将各试样涂敷4周的小鼠背部皮肤的组织学观察结果。图3的(a)部分为确认毛囊数的增加的结果，图3的(b)部分为测量毛囊的直径的结果。各值是5只小鼠的平均±标准误差。ns＝p＞0.05，**＝p＜0.01，***＝p＜0.001(单因素方差分析(one-way ANOVA)，Tukey’s试验)。
图4为示出将各试样涂敷4周的小鼠背部皮肤中毛囊的皮肤表达现象的光学显微镜拍摄结果。
图5为示出涂敷各试样的小鼠背部皮肤组织的β-连环蛋白表达变化。蛋白质印迹(western blot)结果为各组中独立实验的平均±标准误差。(对于各实验n＝2或3)。ns＝p＞0.05，**＝p＜0.01(单因素方差分析(one-way ANOVA)，Tukey’s试验)。
图6为示出涂敷各试样的小鼠背部皮肤组织的基因表达变化。蛋白质印迹结果为各组中独立实验的平均±标准误差。(对于各实验n＝2或3)。ns＝p＞0.05，*＝p<0.05，**＝p＜0.01，***＝p＜0.001(单因素方差分析(one-way ANOVA)，Tukey’s试验)。
具体实施方式
以下，详细说明本发明。
如上所述，除了激素分泌变化以外，压力引起的脱发现象的增加导致可防止脱发和/或促进生发的药物组合物等的需求的增加，然而，在已知对人体安全的来自天然的化合物中，有关可有效防止脱发和/或促进生发的化合物的报告不多。
本发明的壬醛通过调节皮肤组织的生发相关基因及蛋白质的表达来示出脱毛小鼠的背部显示出促进生发效果，因而具有提供对于防止脱发及促进毛发生长有效的组合的效果。
因此，本发明提供包含壬醛或其药学上可接受的盐作为有效成分的防止或治疗脱发用、或者促进生发或育发用药物组合物。
本发明的术语“壬醛(Nonanal)”作为由化学式1表示的烷基醛(alkyl aldehyde)类化合物，国际理论和应用化学联合会(IUPAC)名称为壬醛(nonanal)，还被称为壬醛(pelargonaldehyde)、壬醛(nonanaldehyde)、天然壬醛(nonyl aldehyde)及C-9醛(C-9aldehyde)。壬醛的结构式为C9H18O，分子量为142g/mol。壬醛为无色液体，不溶于水，但可溶于乙醇和丙二醇(propylene glycol)。
化学式1
壬醛作为多种植物的精油成分，被已知为可食用的安全物质，在工业中用于显示出味道和香味。据报告，根据口服毒性实验结果，壬醛的半数致死量(LD50值)在大鼠中为5,000mg/kg。另一方面，据报告，以兔子为对象的经皮毒性结果，LD50值为5,000mg/kg。并且，临床实验结果：1％溶液中未出现皮肤刺激或致敏反应(敏感症)。
另一方面，据报告，以小鼠为对象的实验中，壬醛示出止泻剂活性。在使用硫酸镁，花生四烯酸和PGE2复合物引起腹泻的小鼠中，对蓖麻油经过肠的影响进行评价的结果，蓖麻油的精油成分中壬醛显示出明显的抑制腹泻的效果，且不引起便秘。(Zavala-Sanchezet al.,Antidiarrheal activity of nonanal,an aldehyde isolated from Artemisialudoviciana,Pharmaceutical Biology,40(4)；263-268,2002)。
因此，壬醛在口服给药0.1mg/kg至1,500mg/kg，优选地，口服给药10mg/kg至1,000mg/kg的情况下，其毒性微乎其微，因而可用作化妆品组合物、保健食品组合物、医药品或准药物组合物等。
在本发明中，上述壬醛在与其相同的效果范围内可以包括壬醛水合物、壬醛衍生物等，可以包括其溶剂化合物或立体异构体。
在本发明中，通过阐明上述壬醛的皮肤组织的生发相关基因及蛋白质表达的调节机制，从而确认了壬醛的与防止脱发或促进生发相关的新用途。
本发明的术语“脱发”意味着毛发完全脱离头皮的现象。已开始进行脱发的人具有生长期变短、休止期长的毛发周期，如在以下实施例中所证明，可确认壬醛具有如下的效果，即，使毛发从休止期转变为生长期，在皮肤组织中使与生发及养毛相关的生长因子的表达增加，预防、改善或治疗脱发。
本发明的术语“生发”意味着从头皮生长出毛发，“育发”意味着毛发的长度变长(即，毛发的生长)，与本发明所属技术领域中所使用的另一术语“养毛”用作相同的意思。
本发明的组合物为可增加选自由β-连环蛋白(β-catenin)、T细胞因子(T-cellfactor)、淋巴样增强因子(lymphoid enhancer factor)及细胞周期蛋白D1(cyclin D1)组成的组的蛋白质的表达。具体地，如本发明的实施例2中所示，确认了本发明的组合物增加了在皮肤组织中与涉及使毛囊周期进入生长期的Wnt/β-连环蛋白信号传递体系相关的β-连环蛋白的基因表达，并且可确认作为被认为是促进生发机制的β-连环蛋白和下位信号传递转录因子的T细胞因子及淋巴样增强因子、以及作为靶基因的细胞周期蛋白D1的表达均明显增加。不仅如此，还可以确认与作为目前可促进生发及毛发生长的代表性药物的米诺地尔(米诺地尔)相比，上述蛋白质或基因的表达量明显增加。因此，与现有的合成组合物相比，本发明的组合物可用作对人体无害，并具有在各方面稳定的促进生发及养毛的效果的生发或养毛剂组合物。
优选的，上述壬醛作为皮肤外用组合物，本发明的包含壬醛作为有效成分的皮肤外用药物组合物作为防止脱发及改善、促进生发用皮肤外用剂，可通过制备成乳膏、凝胶、贴剂、喷雾剂、软膏剂、硬膏剂、洗剂、搽剂、糊剂及泥敷剂的皮肤外用剂形态的药物组合物来使用，但并不局限于此。
本发明的壬醛可用作药学上可接受的盐的形态，作为盐有用的是由药学上可接受的游离酸(free acid)形成的酸加成盐。酸加成盐可通过盐酸、硝酸、磷酸、硫酸、氢溴酸、氢碘酸、亚硝酸或亚磷酸等的无机酸类和脂肪族单体及二羧酸(Dicarboxylate)、苯基取代的链烷酸酯(Alkanoate)、羟基脂肪酸酯(Hydroxyalkanoate)及链烷二酸酯(Alkanedioate)、芳香族酸类、脂肪族磺酸及芳香族磺酸类等的无毒性有机酸获取。这种药学上无毒的盐类包括硫酸盐(sulfate)、焦硫酸盐(pyrosulfate)、硫酸氢盐(bisulfate)、亚硫酸盐(sulfite)、亚硫酸氢盐(bisulfite)、硝酸盐(nitrate)、磷酸盐(phosphate)、磷酸一氢盐(monohydrogenphosphate)、磷酸二氢盐(dihyrogenphosphate)、偏磷酸盐(metaphosphate)、焦磷酸盐(pyrophosphate chloride)、溴化物(bromide)、碘化物(iodide)、氟化物(fluoride)、醋酸盐(acetate)、丙酸盐(propionate)、癸酸盐(decanoate)、辛酸盐(caprylate)、丙烯酸酯(acrylate)、甲酸酯(formate)、异丁酸酯(isobutyrate)、癸酸酯(caprate)、庚酸酯(heptanoate)、丙炔酸盐(propiolate)、草酸盐(oxalate)、丙二酸盐(malonate)、琥珀酸盐(succinate)、辛二酸盐(suberate)、癸二酸酯(sebacate)、富马酸酯(fumarate)、马来酸酯(maleate)、丁炔-1,4-二酸酯(butyne-1,4-dioate)、己烷-1,6-二酸(hexane-1,6-dioate)、苯甲酸(benzoate)、氯苯甲酸(chlorobenzoate)、苯甲酸甲酯(methyl benzoate)、二硝基苯甲酸盐(dinitrobenzoate)、羟基苯甲酸盐(hydroxybenzoate)、甲氧基苯甲酸盐(methoxybenzoate)、邻苯二甲酸盐(phthalate)、对苯二甲酸盐(terephthalate)、苯磺酸盐(benzene sulfonate)、甲苯磺酸盐(toluene sulfonate)、氯苯磺酸盐(chlorobenzene sulfonate)、二甲苯磺酸盐(xylenesulfonate)、苯乙酸盐(phenylacetate)、丙酸苯酯(Phenyl propionate)、丁酸苯酯(phenyl butyrate)、柠檬酸盐(citrate)、乳酸盐(lactate)、β-羟基丁酸酯(β-hydroxybutyrate)、乙醇酸酯(glycolate)、马来酸酯(maleate)、酒石酸酯(tartrate)、甲磺酸盐(methane sulfonate)、丙磺酸盐(propane sulfonate)、萘-1-磺酸盐(naphthalene-1-sulfonate)、萘-2-磺酸盐(naphthalene-2-sulfonate)或扁桃酸盐(mandelate)。
根据本发明的酸加成盐可通过常规方法制备，即，例如，将上述壬醛溶解在过量的酸性水溶液中，使用例如甲醇、乙醇、丙酮或乙腈等的水混溶性有机溶剂来使上述盐沉淀。还可以通过如下的方法制备，即，对相同量的壬醛及水中的酸或醇进行加热，接着蒸发或干燥上述混合物或抽滤析出的盐。
并且，可使用碱来制备药学上可接受的金属盐。碱金属或碱土金属盐通过如下的方式获得，即，例如将化合物溶解在过量的碱金属氢氧化物或碱土金属氢氧化物溶解中，过滤不溶性化合物盐，并蒸发干燥滤液。在这种情况下，金属盐适合制备钠盐，钾盐或钙盐。并且，相应的银盐是通过碱金属或碱土金属盐与适当的银盐(例如硝酸银)反应获得的。
并且，本发明的上述壬醛不仅包括药学上可接受的盐，还包括可通过常规方法来制备的盐、水合物及溶剂化物。
根据本发明的加成盐可通过常规的方法来制备，例如，将上述壬醛溶解在丙酮、甲醇、乙醇或乙腈等的水混和性的有机溶剂中，添加过量的有机酸或无机酸的酸水溶液，使其沉淀或结晶。接着，从上述混合物蒸发溶剂或过量的酸后使其干燥，或者抽滤析出的盐。
当本发明的组合物用作医药品的情况下，包含壬醛或其药学上可接受的盐作为有效成分的药物组合物在临床给药时，可通过制剂化为以下口服或非口服给药形态的方式来给药，但并不局限于此。
作为口服给药用剂型包括片剂、丸剂、轻/软胶囊剂、液体、悬浮液剂、洗剂剂、糖浆剂、颗粒剂、酏剂等，这些剂型除了有效成分以外，还包含稀释剂(例如，乳糖、右旋糖、蔗糖、甘露醇、山梨糖醇、纤维素和/或甘氨酸)、润滑剂(例如，二氧化硅、滑石、硬脂酸及其镁或钙盐和/或聚乙二醇)。片剂可包含硅酸铝镁、淀粉糊、明胶、甲基纤维素、羧甲基纤维素钠和/或聚乙烯吡咯烷酮等的结合剂，根据情况可包含淀粉、琼脂、海藻酸或其钠盐等的崩解剂或沸腾混合物和/或吸收剂、着色剂、调味剂以及甜味剂。
本发明的壬醛或其药学上可接受的盐作为有效成分的药物组合物能够以非口服的方式给药，非口服给药的方法包括皮下注射、静脉注射、肌肉内注射或胸部内注射。在这种情况下，为了制剂化为非口服给药用剂型，将上述壬醛或其药学上可接受的盐与稳定剂或缓冲剂混合在水中，以制备溶液或悬浮液，其可以制备成安瓿或小瓶单位的给药型。上述组合物可包含已灭菌/可灭菌的防腐剂、稳定剂、可湿性粉剂或乳化促进剂、用于调节渗透压的盐和/或缓冲剂等的添加剂、以及其他治疗上有用的物质，并且可以通过作为常规方法的混合、科利华或涂敷方法来制剂化。
并且，本发明的化合物对人体的给药量可根据患者的年轻、体重、性别、给药形态、健康状态及疾病严重程度而变化，以体重为60㎏的成人患者为基准，通常为0.001～1,000㎎/日，优选地，为0.01～500㎎/日，并且可以按照医生或药剂师的判断间隔规定时间以一天一次至数次的方式分剂量给药。
并且，本发明提供包含壬醛或其药学上可接受的盐作为有效成分的防止或改善脱发用、或者促进生发或育发用准药物。上述壬醛的具体内容如上所述。
在将本发明的壬醛用作准药物组合物的有效成分的情况下，可以原样加入呈现出防止脱发及促进毛发生长效果的上述壬醛，或与其他准药物或准药物成分一起使用，可按照常规方法适当的使用。有效成分的混合量可根据使用目的适当确定。
本发明的防止脱发及促进毛发生长用准药物组合物的剂型没有特别限制，可以剂型化为本发明所属技术领域已知的准药物的形态，以呈现防止脱发及促进毛发生长的效果。上述剂型化的准药物包括头皮滋补剂、头皮洗剂、头皮乳膏、头皮乳清、头皮精华液、头皮浓缩精华、头皮护理、头皮护理剂、头皮洗发剂、头皮膜、头发滋养剂、头发洗剂、头发乳膏、喷发剂、发用摩丝、发胶、护发素、洗发水、润发液、发膜、焗油膏、眉毛生长剂、睫毛生长剂、睫毛营养剂、宠物用洗发水、宠物用润发液、洗手液、洗剂皂、香皂、消毒清洁剂、湿纸巾、面膜、软膏剂、贴剂或过滤填料等，并包括常规意义上的所有准药物。
并且，在各剂型中，防止脱发及促进毛发生长用准药物组合物可以将其他成分根据其他准药物的剂型或用途等任意选定并配合。有效成分的混合量可根据用途适当地确定，可包括，例如，增稠剂、稳定剂、增溶剂、维生素、颜料及调味剂等的常规添加剂、及载体等。
以总重量为基准，优选地，上述组合物的含量分别为0.0001重量百分比至10重量百分比。在大于10重量百分比的情况下，制备组合物时颜色及稳定性下降，在小于0.0001重量百分比的情况下，存在其效果微乎其微的缺点。如在治疗指数上所确认，本发明的包含壬醛作为有效成分的准药物组合物对于细胞几乎没有毒性及副作用，因此可用作准药物材料。
并且，本发明提供包含壬醛或其药学上可接受的盐作为有效成分的防止或改善脱发用、或者促进生发或育发用化妆品组合物。
除了作为有效成分的壬醛或其药学上可接受的盐以外，本发明化妆品组合物中所包含的成为还包含化妆品组合物中常用的成分，例如，抗氧化剂、稳定剂、增溶剂、维生素、颜料及调味剂等的常规添加剂、及载体等。
本发明的化妆品组合物可以制备为本发明所属技术领域常规制备的任何剂型，并可以被制剂化为溶液、悬浮液、乳浊液、糊剂、凝胶、乳膏、洗剂、粉末、肥皂、含表面活性剂的清洁剂、油、粉底、乳浊液粉底、蜡粉底及喷雾等，但并不局限于此。
在本发明的剂型为糊剂、乳膏或凝胶的情况下，作为载体组分可使用动物油、植物油、蜡、石蜡、淀粉、黄芪胶、纤维素衍生物、聚乙二醇、硅氧烷、膨润土、二氧化硅，滑石或氧化锌等。
在本发明的剂型为粉末或喷雾剂的情况下，作为载体组分可使用乳糖、滑石、二氧化硅、氢氧化铝、硅酸钙或聚酰胺粉末，尤其，在喷雾剂的情况下，还可以包括氯氟烃，丙烷/丁烷或二甲醚等的推进剂。
在本发明的剂型为溶液或乳浊液的情况下，作为载体组分使用溶剂、增溶剂或增溶剂，例如包括水、乙醇(ethanol)、异丙醇(isopropanol)、碳酸乙酯(ethyl carbonate)、乙酸乙酯(ethyl acetate)、苯甲醇(benzyl alcohol)、苯甲酸苄酯(benzyl benzoate)、丙二醇(propylene glycol)、1,3-丁二醇油(1,3-butyl glycol oil)、甘油脂肪族酯(glycerol aliphatic ester)、聚乙二醇(polyethylene glycol)或脱水山梨糖醇脂肪酸酯。
在本发明的剂型为悬浮液的情况下，作为载体组分可使用水、乙醇或丙二醇等的液体稀释剂、乙氧基化异硬脂醇、聚氧乙烯山梨糖醇酯及聚氧乙烯脱水山梨糖醇酯等的悬浮剂、微晶纤维素、偏氢氧化铝、膨润土，琼脂或黄芪胶等。
在本发明的剂型为含表面活性剂的清洁剂的情况下，作为载体组分可使用脂肪醇硫酸盐、脂肪醇醚硫酸盐、磺基琥珀酸单酯、羟乙基磺酸盐、咪唑啉鎓衍生物、甲基牛磺酸盐、肌氨酸盐、脂肪酸酰胺醚硫酸盐、烷基酰胺基甜菜碱、脂肪醇、脂肪酸甘油酯、脂肪酸二乙醇酰胺、植物油、羊毛脂衍生物或乙氧基化甘油脂肪酸酯等。
并且，本发明提供包含壬醛或其药学上可接受的盐作为有效成分的防止或改善脱发用、或者促进生发或育发用保健食品组合物。
根据本发明的食品组合物可以以多种形态根据本发明所属技术领域已知的常规方法来制备。常见的食品虽不限于此，但可通过在饮料(包括含酒精饮料)、水果及其加工食品(例如，水果罐头、瓶装罐头、果酱、柑橘属果酱等)、鱼类、肉类及其加工食品(例如，火腿、香肠、腌牛肉等)、面包类、面类(例如，乌冬面、荞麦面、拉面、意大利面、通心粉等)、果汁、各种饮料、饼干、饴糖、乳制品(例如，黄油、芝士等)、食用植物油、人造黄油、植物性蛋白、蒸煮食品、冷冻食品、各种调味料(例如，大酱、酱油、沙司等)等加入本发明的壬醛来制备。并且，作为营养添加剂虽不限于此，但可通过对胶囊、片剂、丸剂等加入本发明的壬醛来制备。并且，作为保健食品虽不限于此，例如，通过液化、颗粒化、胶囊化及粉末化来将本发明的壬醛以茶、果汁及饮料的形态制备，使得其以饮用(保健饮料)的方式被摄入。并且，为以食品添加剂的形态使用本发明的壬醛，可以以粉末或浓缩液的形态制备。并且，本发明的壬醛可以通过与具有防治脱发及促进生发效果的已知的活性成分混合来以组合物的形态制备。
在本发明的壬醛用作保健饮料的情况下，与常规饮料相同地，上述保健饮料组合物可包含各种风味剂或天然碳水化合物等作为附加成分。上述天然碳水化合物可以使用：葡萄糖、果糖等的单糖(monosaccharide)；麦芽糖、蔗糖等的二糖类；糊精、环糊精等的多聚糖(polysaccharide)；木糖醇、山梨糖醇、赤藓糖醇等的糖醇。甜味剂可以使用：奇异果甜蛋白(thaumatin)、甜叶菊提取物等的天然甜味剂；糖精、阿司帕坦(aspartame)等的合成甜味剂等。上述天然碳水化合物的比率为，每100mL本发明的组合物通常为约0.01～0.04g，优选地，约0.02～0.03g。
并且，本发明的壬醛可作为防止脱发及促进生发用保健食品的有效成分来包含，其量不受特别限制，为用于实现防止脱发及促进毛发生长的效果的量。对于整体组合总重量，优选地，为0.01重量百分比至100重量百分比。本发明的食品组合物可以通过将壬醛和已知的具有防止脱发及促进毛发生长效果的其他活性成分混合来制备。
除上述之外，本发明的保健食品可包括各种营养剂、维生素、电解质、风味剂、着色剂、果胶酸、果胶酸盐、海藻酸、海藻酸盐、有机酸、保护胶体增稠剂、pH调节剂、稳定剂、甘油、醇或碳酸化剂等。此外，本发明的保健食品还可以包括天然果汁、果汁饮料、或者用于蔬菜饮料的制造的果肉。这种成分可以单独使用或混合使用。这种添加剂的比率虽不重要，但通常本发明的组合物在每100重量份中选择0.01～0.1重量份的范围。
以下，参考实施例详细说明本发明。然而，这些实施例用于进一步具体说明本发明，本发明的范围不限于这些实施例。
<实施例>
实施例1.壬醛的促进生发效果
1-1.实验动物的饲养及生发剂的涂敷
(1)试样的准备
载体(Vehicle)(乙醇：水：丙二醇＝5：3：2)和作为对照药物的米诺地尔(MXD)、以及作为试验物质的壬醛(NN)均购自西格玛奥德里奇(Sigma Aldrich)(株)公司。
(2)实验动物的饲养
从Orientbio(株)购买24只出生6周龄的C57BL/6N雄鼠，在饲养室环境中适应两个星期后，分为阴性对照组(CON)、阳性对照组(MXD)以及壬醛组(NN)供三个组(n＝5)来进行实验。动物饲养室维持了温度21±2.0℃、相对湿度50±5％、12小时昼夜周期环境。实验期间，给小鼠自由供给常规固体饲料(chow)和水。
(3)生发剂的皮肤涂敷及肉眼观察方法
为了观察促进生发效果，使用具有背部皮肤的颜色呈粉红色(pink)的休止期体毛的8周龄小鼠。使用小鼠用剪毛器(clipper)剔掉小鼠背部的毛后，每天下午四点涂敷试样一次，持续四周。将作为实验物质的壬醛(3mg/小鼠/日)和作为对照药物使用的米诺地尔(3mg/小鼠/日)溶解在载体(乙醇：水：丙二醇＝5：3：2)来使用，并对阴性对照组只涂敷载体。
为了肉眼确认毛生长的状态而开始涂敷，在经过一周、两周、三周以及四周的时间点利用乙醚轻度麻醉实验动物后，拍摄背部照片。使用尺子来测量毛的长度，以评估毛生长的程度。
1-2.体重变化的测量
在涂敷生发剂之前到结束的时间点期间每周测量实验动物的体重。阴性对照组(CON组)和实验组(NN组)以及阳性对照组(MXD组)的初始体重无显著差异，在涂敷实验物质四周后测量的体重也无显著差异。
1-3.生发状态及毛的长度的变化
(1)生发的肉眼特征
将试样涂敷在小鼠的背部部位四周，通过拍摄照片来确认每周生长毛的形态。当进入休止期(telogen)的小鼠剃除毛时，体表面的颜色呈粉红色。在随着实验的进行体表面的颜色变色黑色的情况下，意味着毛发周期从休止期进入生长期(anagen)。在阴性对照组(CON)的情况下，三周后毛几乎没有生长，在壬醛涂敷组(NN)中，毛从涂敷第三周开始生长，与阴性对照组相比，四周后观察到在更多的个体中均匀且旺盛地生长出毛(图1)。
(2)毛长度的变化
在实验期间(四周)每周测量毛的长度。与阴性对照组相比，NN组的毛的长度从第三周开始显著增加，四周后显著增加至434％(p＜0.001)(图2)。因此，可知壬醛具有优异的促进毛发的生长的效果。
1-4.背部皮肤的组织学分析
涂敷四周后，处死实验动物，利用剪刀和钳子提取以涂敷试验物质的部位为中心的皮肤组织试样后，用福尔马林(formalin)固定。利用醇和二甲苯(xylene)逐步脱水后，将其包埋在石蜡中，用切片机制造5μm的切片，然后用醇和二甲苯除去石蜡。实施苏木精-伊红(Hematoxylin-Eosin)染色后，利用光学显微镜观察毛囊组织的组织学变化。病理镜检负责人通过镜检评估生发周期，在光学显微镜上测量毛囊(hair follicle)的数量及直径，在40倍率光学显微镜下测量了各试验组的毛囊数，其结果显示在图3及图4。
其结果，如图3所示，可知与阴性对照组(CON)相比，NN组(增加166％)和MXD组的情况下，毛囊数显著增加，通过图像分析评估各试验组的毛囊直径的结果，与阴性对照组相比，在NN组(增加38％)和MXD组的毛囊直径显著增加。并且，如图4所示，利用显微镜测量的结果，观察到在NN组和MXD组中毛囊延长并表露到皮肤的现象，这种现象认为与两个物质的促进生发效果有关。
实施例2.基于壬醛调节小鼠的皮肤组织的生发相关基因及蛋白质表达
2-1.分析皮肤组织中与促进生发相关的蛋白质的表达
(1)蛋白质印迹(Western blot)分析法
将规定量的皮肤组织用液氮和裂解缓冲液(lysis buffer)在研钵中均质化，然后在13,000×g，4℃下离心分离20分钟，然后取中间层并借助考马斯亮蓝(又称Bradford)法蛋白质定量。将50μg的蛋白质在SDS-聚丙烯酰胺凝胶(SDS polyacrylamide gel)上电泳，然后电印迹(electroblotting)到聚偏二氟乙烯(PVDF)超薄膜，并分别与β-连环蛋白及β-肌动蛋白抗体(Cell Signaling Technology公司，Danvers,MA,USA)反应。用化学发光检测系统(chemiluminescent detection system,Amersham)使各蛋白质的信号可视化，然后使用Quantity One Analysis Software(Bio-Rad Laboratories)定量化条带厚度。
(2)与促进生发相关的蛋白质的表达分析结果
已知Wnt/β-连环蛋白信号传递系统是参与通过调节毛囊周期来促进进入生长期(anagen)的信号传递系统。为了确认这些与促进生发相关的信号传递系统是否活性化，借助蛋白质印迹在背部皮肤组织分析β-连环蛋白表达量的结果，在NN组的情况下，与阴性对照组相比，β-连环蛋白的蛋白质表达显著增加71％，这种效果比米诺地尔组优秀(图5)。
2-2.分析在皮肤组织中与促进生发相关的基因表达
(1)利用Trizol方法的核糖核酸(RNA)分离
为了从实验动物的皮肤组织分析定量总核糖核酸，将1mL的Trizol溶液放入皮肤组织50～100mg并均质化，然后在12,000×g，4℃下离心分离10分钟。将上清液转移到新管中，然后加入200μl氯仿(chloroform)，剧烈摇动。将该过程重复两次，然后将上清液转移到新管中，并以1：1的比率加入异丙醇(isopropanol)和上清液。剧烈摇动10次，然后在室温放置10分钟，然后在12,000×g，4℃下离心分离10分钟，以去除上清液，将70％1ml的乙醇加入剩余的沉淀物，然后在7,500×g，4℃下离心分离5分钟。除去乙醇后，将装有核糖核酸沉淀物的试管在室温下干燥5分钟，用无核酸酶水(nuclease free water)溶解核糖核酸沉淀物(pellet)。使用UV/VIS分光光度计(spectrophotometer，Beckman coulter，DU730)测量在260nm及280nm波长下提取的核糖核酸试样的浓度，并进行琼脂糖凝胶电泳以确认核糖核酸试样的完整性(integrity)。
(2)定量聚合酶链锁反应(q-PCR，Quantitative-Polymerase Chain Reaction)方法
分离核糖核酸后，使用反应试剂盒和Superscript逆转录酶(reversetranscriptase，Gibco BRL，Gaithersburg，MD，USA)来进行逆转录，从而合成互补脱氧核糖核酸(cDNA)。为了测量基因的表达量，利用荧光染料掺入法(SYBR Green)(Bio-Rad,Hercules，CA，USA)进行实时定量即时聚合酶链锁反应(PCR，Real-time q-PCR)，仪器使用了real-time PCR(CFX ConnectTMReal-Time PCR Detection System,Bio-Rad)。与各个基因有关的聚合酶链式反应(PCR)引物的碱基序列如表1所示。
为了实时聚合酶链式反应，将2μL的互补脱氧核糖核酸、10μL的2X SYBR mix、1μL的分别为10～20pmol/μL的正向引物(forward primer)和反向引物(reverse primer)加入20μL的总体积，其余的都充填了H2O。聚合酶链锁反应的扩增步骤如下，扩增循环进行了35循环。为了热起动(Hot start)在95℃下进行1分钟，在95℃下进行扩增步骤的变形(denaturation)15秒，在55℃下热处理(annealing)30秒，在72℃下扩增(extension)1分钟。在所有循环完成后，实施了熔解曲线(melting curve)分析以确认引物的特异性。
表1
用于定量聚合酶链锁反应的引物序列。
(3)与促进生发先关基因的表达的调节分析结果
借助定量聚合酶链锁反应分析法分析涂敷了4周实验物质的小鼠的背部皮肤组织上的被称为促进生发机制的β-连环蛋白信号传递物质的表达变化。
其结果也如图6所示，在NN组的情况下，与阴性对照组相比，β-连环蛋白和下位信号传递转录因子(T细胞因子及淋巴样增强因子)以及作为靶基因的细胞周期蛋白D1的表达均显著增加。因此，可知壬醛得促进毛发生长效果可通过β-连环蛋白信号传递活性化显示。
序列表
<110> 延世大学校产学协力团
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Claims (7)
1.一种预防或治疗脱发用、或者促进生发或育发用药物组合物，其特征在于，包含壬醛或其药学上可接受的盐作为有效成分。
2.根据权利要求1所述的预防或治疗脱发用、或者促进生发或育发用药物组合物，其特征在于，上述组合物使选自由β-连环蛋白、T细胞因子、淋巴样增强因子及细胞周期蛋白D1组成的组中的蛋白质的表达增加。
3.根据权利要求1所述的预防或治疗脱发用、或者促进生发或育发用药物组合物，其特征在于，作为皮肤外用剂，上述药物组合物为选自由乳膏、凝胶、贴剂、喷雾剂、软膏剂、硬膏剂、洗剂、搽剂、糊剂及泥敷剂组成的组中的剂型。
4.一种防止或改善脱发用、或者促进生发或育发用准药物组合物，其特征在于，包含壬醛或其药学上可接受的盐作为有效成分。
5.一种防止或改善脱发用、或者促进生发或育发用化妆品组合物，其特征在于，包含壬醛或其化妆品学上可接受的盐作为有效成分。
6.根据权利要求5所述的防止或改善脱发用、或者促进生发或育发用化妆品组合物，其特征在于，上述化妆品组合物为头皮滋补剂、头皮洗剂、头皮乳膏、头皮乳清、头皮精华液、头皮浓缩精华、头皮护理、头皮护理剂、头皮洗发剂、头皮膜、头发滋养剂、头发洗剂、头发乳膏、喷发剂、发用摩丝、发胶、护发素、洗发水、润发液、发膜、焗油膏、眉毛生长剂、睫毛生长剂、睫毛营养剂、宠物用洗发水或宠物用润发液的剂型。
7.一种防止或改善脱发用、或者促进生发或育发用保健食品组合物，其特征在于，包含壬醛或其食品学上可接受的盐作为有效成分。
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