CN100337685C - 减少毛发生长、毛囊及毛干尺寸与毛发色素沉着 - Google Patents
减少毛发生长、毛囊及毛干尺寸与毛发色素沉着 Download PDFInfo
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Abstract
本发明利用天然及/或合成丝氨酸蛋白酶抑制剂或含丝氨酸蛋白酶抑制活性之植物萃取物,添加或不添加一或多种异黄酮及/或含一或多种异黄酮之额外的天然萃取物,它们能影响哺乳动物毛发生长、毛囊及毛干尺寸与改变毛发色素沉着。
Description
本发明有关一种可有效地减缓毛发生长之方法及组合物。详言之,本发明有关一种改变毛发生长速率、缩小毛囊及毛干尺寸、及减少毛干色素沉着之方法,其系局部施用含有丝氨酸蛋白酶抑制活性之植物萃取物,尤其是大豆萃取物诸如豆浆,或前述物质之混合物及调配物,同时结合以其他活性成分诸如异黄酮。
哺乳类毛发的一种主要功能系提供外在保护。然而,在人类身上几乎已完全丧失此种功能,其毛发基本上系基于社交及美观因素而保持或移除。
已使用许多方法以移除不需要之毛发,包括剃除、电针除毛、拔除、激光及光疗及注射治疗性抗雄激素物质。此等习用方法不无缺点。例如,剃除可能于皮肤表面产生割痕及刀痕,可能明显地增加毛发再生长速率,亦可能留下不期望产生之短粗毛。虽然电针除毛可长时间使某一区域保持没有不期望之毛发,但此种方法经常是昂贵而疼痛,亦可能导致疤痕。拔除不仅会导致疼痛及不适,亦经常无法完全移除短毛。使用抗雄性经常产生数种不期望之副作用,诸如影响肌肉之形成。因此,需要一种可减缓毛发生长之较佳方法。
假性须囊炎系为发炎性毛发疾病,大多发生于胡须区域。当毛发尖端穿入皮肤而非穿透毛孔时,则产生发炎性小囊丘疹。此种过程于黑种人身上极为常见,因其毛发通常卷曲,而于锐角下穿出皮肤。密贴式的剃除,尤其是使用剃刀,易罹患假性须囊炎。目前最有效之治疗系使该等毛发完全于皮肤表面上生长。但经常不期望使用该种治疗。
多毛症系为极为常见之疾病,约有4%女性患有此症。颜面多毛症经常防碍个人及工作活动,而暂时性的毛发移除系为处理多毛患者的主要方式。剃除系为颜面毛发最常使用之暂时方法,因为拔除、敷蜡及使用除毛剂更难以忍受,且需谨慎以避免毛囊炎、色素沉着、及产生疤痕。通常使用化妆品覆盖以遮蔽刀痕及短毛,而电针除毛及热除可适当地用于永久性除毛。
除毛之取代性或互补式研究系为减缓毛发生长、缩小毛囊及毛干尺寸及减少毛干色素沉着之方法。该项研究可降低现存毛发之可见度,使其变得较细较柔软。与其他除毛法结合时,该方法可促进并延长除毛效果,并降低除毛之需求及频率。长期使用该项研究结果,可使毛发生长变稀、变软而降低色素沉着,使其变得较不易察觉,而不需使用其他除毛方法。
腋部(腋窝)需减缓毛发生长,此处使用除臭剂及制汗剂以控制腋毛内所产生之气味。期望有可供腋下使用之产品,在除臭或制汗活性之同时具有减缓毛发生长活性。
非裔型毛发的形态独特--具有不同直径之卷曲毛干。此种复杂之毛干结构需要特殊之修饰产品及方法,以确定非裔型毛发保持美观上所需之性质。期望有一种产品,可降低此种复杂性,使该非裔型毛发更易处理,并改善其外观。
期望提供一种方法,以化学或自然地影响毛发生长、毛囊及毛干尺寸及毛干色素沉着,而不对使用者产生不需要之副作用。
根据本发明,吾人已发现一种组合物及方法,以改变哺乳类毛发生长、毛囊及毛干尺寸及毛发色素沉着,其系于哺乳类皮肤上局部施用有效量之局部活性组合物,包含蛋白酶抑制剂、植物萃取物,尤其是豆科植物萃取物包括但不限于豆浆,历经足以影响毛发生长、毛囊及毛干尺寸及毛干色素沉着之时间。该种局部活性组合物可另外组合以其他活性成分,包括但不限于合成或天然异黄酮,以促进针对毛发生长及色素沉着所期望之效果。
本发明组合物及方法提供一种独特而简便之方式,以减缓毛发生长、缩小毛囊及毛干尺寸并减少毛干色素沉着,其系使用含有丝氨酸蛋白酶抑制活性之丝氨酸蛋白酶抑制剂、豆科植物之植物萃取物,尤其但不限于豆浆,及其与异黄酮之组合物。
附图简单说明
本发明含有至少一种彩色图型。此种具有彩色图型之专利的副本系于请求及付出必要费用下由专利商标局提供。
参照以下本发明详述及附图可进一步明了本发明及其他优点,其中:
图1:对照组及经豆浆处理之C57B1/6老鼠毛发的相片(高倍率)。
图2:对照组及经豆浆处理之C57B1/6老鼠毛干于毛发周期第四日之组织剖面图。
图3:对照组及经豆浆处理之C57B1/6老鼠毛干于毛发周期第七日之组织剖面图,高及低倍率。
图4:对照组及经豆浆处理之C57B1/6老鼠毛干于毛发周期第18日之组织剖面图。
图5:对照组及经豆浆处理之C57B1/6老鼠毛干于毛发周期第21日之组织剖面图。
图6:对照组、经豆浆处理及经豆浆衍化蛋白质处理之C3H老鼠毛干(高倍率)于毛发周期第七日之组织剖面图。
图7:对照组、经豆浆处理及经豆浆衍化蛋白质处理之C3H老鼠毛干(低倍率)于毛发周期第七日之组织剖面图。
图8:对照组、经豆浆处理及经豆浆衍化蛋白质处理之C3H老鼠毛干于毛发周期第21日之组织剖面图。
图9:对照组及经豆浆衍化蛋白质处理之C3H老鼠毛发之相片(高倍率)。
图10:对照组及经豆浆衍化蛋白质处理之C57B1/6老鼠毛干(高倍率)于毛发周期第八日之组织剖面图。
图11:说明豆浆之色胺酸抑制活性的图。
图12:C57B1/6老鼠皮肤于整个毛发周期之Western印迹法,说明于豆浆处理后降低之酪胺酸及TRP-1蛋白质浓度。
图13:人类颜面未经处理及经豆浆处理侧面的相片,每日使用豆浆处理历经四周。
图14:使用及不使用豆浆处理时毛干尺寸定量分析。
图15:于一腿部使用豆浆处理五周之后的人类腿部毛发相片。
图16:对照组、经豆浆处理、及经富含异黄酮之豆浆处理之C57B1/6老鼠毛发(高倍率)的相片。
图17:对照组、经豆浆处理、及经富含异黄酮之豆浆处理之C57B1/6老鼠皮肤于毛发周期第15日之组织剖面图,说明毛干之厚度及颜色。
图18:使用各种豆浆及异黄酮调配物处理三周后,C57B1/6老鼠毛发之相片。
较佳具体实施例详述
本发明中,″哺乳类″应包括任何″高级脊椎动物包括哺乳动物类″,如Webster氏医学字典407(1986)所定义,包括但不限于人类。本发明所使用之″(%,重量/体积)″意指每100毫升总组合物之特定成分克数。
适用于本发明组合物之局部活性剂系包括蛋白酶抑制剂及具有蛋白酶抑制活性之天然植物萃取物及其混合物。较佳蛋白酶抑制剂系为丝氨酸蛋白酶抑制剂,尤其是大豆色胺酸抑制剂(″STI″)及由大豆衍生之Bowman Birk抑制剂(″BBI″)。较佳植物萃取物系为豆科植物者,尤其是大豆萃取物,诸如豆浆。该蛋白酶抑制剂以本发明组合物总体积计之较佳用量系由约0.0001%(重量/体积)至约20%(重量/体积),更佳系由约0.001%(重量/体积)至约5%(重量/体积)。较佳之植物含水萃取物诸如豆浆之含量系10-99%(体积/体积),更佳系由50-99%(体积/体积)。
吾人已意外地发现当局部活性剂诸如前述者--尤其是豆浆或含有豆浆之调配物--富含异黄酮时,尤其是由大豆衍化之异黄酮,可增进其对于毛发生长、毛发尺寸及毛发色素沉着之抑制效果。该植物含水萃取物诸如豆浆中之异黄酮含量较佳系0.000005-15%(体积/体积),更佳系0.00001-10%(体积/体积)。
若需要该局部活性医药或化妆剂之输送参数,则本发明局部活性组合物可另外包含药学上可接受或化妆学上可接受之媒质,其可作为输送系统,以使该局部活性剂渗入毛干及皮肤内。
该医药或化妆组合物可视情况结合其他成分,诸如润湿剂、化妆佐剂、抗氧化剂、脱色剂、抗老化剂、除毛剂、头发造型剂、防晒剂、表面活性剂、发泡剂、调理剂、保湿剂、香料、着色剂、稠化剂、缓冲剂、防腐剂等,及其混合物。可于不影响该丝氨酸蛋白酶活性之用量下结合此等物质,以产生化妆或医药产品,诸如但不限于精油、乳霜、洗剂、糊剂、凝胶、粉末、敷剂或注射物,以减缓毛发生长、毛发尺寸及毛发色素沉着。
本发明组合物可于其他活性成分或组合物之前、同时或之后施用,以促进其效果。例如,本发明组合物可与一或多种有利于除毛之产品结合使用,以实际移除毛发,降低毛发可见度、改善毛发风采或改善毛发处理。本发明组合物可于除毛之前、之期间或之后施用。其可与下列物质中之一或多种同时局部施用:除毛剂、洗发精、毛发调理剂、造型凝胶、护发产品、发蜡产品、剃毛产品、除毛产品、剃毛后产品、除臭剂、制汗剂、电针除毛、激光除毛、光导性除毛、敷剂或沐浴添加剂。
本发明组合物可每日施用至少历经四至八周,如此应可发现其对毛发外观之影响。可连续施用,先决条件为期望保持该毛发之状态。每日施用于脸部可缓和假性须囊炎及/或多毛症之状况;施用于腋部可减缓腋下之毛发生长,施用于头皮及头发上有助于非裔型毛发之处理及造型。
该局部活性医药或化妆组合物之施加量应可有效地改变哺乳类毛发生长、毛囊及毛干尺寸及毛干色素沉着。本发明所使用之″有效量″意指足以覆盖皮肤表面期望延迟毛发生长及毛发色素沉着及缩小毛发尺寸之区域之量。当期望延迟毛发生长、毛发尺寸及毛发色素沉着时,该组合物较佳系施加于皮肤表面,使得基于每平方厘米皮肤表面存有约2微升/平方厘米至约500微升/平方厘米之局部活性剂。
吾人意外地发现当局部活性剂诸如豆浆或富含异黄酮之豆浆局部施加于动物皮肤时,明显延迟毛发生长、毛囊及毛干尺寸及毛干色素沉着。吾人亦相信因为人类之毛发生长周期经常较老鼠缓慢,故于人类体内之毛发生长延迟远长于老鼠。
本文所述的发明可适当地于任何组份、成分、或未详细揭示于本发明之步骤等不存在下进行。下文阐述的实施例用于进一步说明本发明之性质及其进行方式。然而,本发明应不限于其细节。
实施例
实施例1:试验物质于老鼠系统中之除毛性
C57BI/6或C3H老鼠(雄性及雌性)系得自Charles River(Kingston,NY),8-10周大,处于其个别毛发周期之休止(静止)期。根据Stenn等人″糖皮质激素对毛发生长起始之影响:再审查″,6 Skin Pharmacol.,125-134(1993)所列示之方法,通过敷蜡除毛(拔除)动物的各背毛而诱发毛发生长。8-10周大之C57B1/6及C3H老鼠于除毛时所有毛囊皆同时开始生长期(毛发生长期)。如表1所示,于诱发部位发现以下现象:
表1:于诱发部位之发现
诱导后之日数 | 诱导部位之形态及组织发现 |
1-2(前生长期) | 开始生长新毛囊 |
3至4 | 毛囊完全发展,但其毛干尚未可见 |
7至8(后生长期) | 每只老鼠皆具有极黑之皮肤;组织上可见其毛干 |
11-12 | 毛干开始穿透表皮 |
14 | 每只老鼠皆覆有短毛 |
19 | 组织上发现毛囊退化(生长中期) |
21-25 | 毛囊退化至休止期 |
如表1所示,当除毛后数日动物粉红色皮肤开始变黑时,可见到毛发生长。此因该C57BI/6及C3H老鼠仅于毛囊中含有黑素细胞,而背部表皮中则否,故毛干中有毛发色素沉着。使用市售产品进行化学除毛时,吾人国际申请案编号PCT/US97/11033中记录了相似的毛发生长特性。
因为鼠类毛发周期不仅因种类不同而异,个别动物之间亦有差异,研究时于每只动物体内分析毛发周期之状态。使用剪刀自每只老鼠身上取下2厘米×1厘米皮肤试样,于25℃下使用pH约6.9-7.1之10%经缓冲甲醛水溶液(StephensScientific)固定,根据已知方法成型于石蜡块中。该蜡块经切薄片,剖面使用H&E或Fontana-Mason染色法染色。组织性地检测该剖面,使用本领域众所周知之方法,以确定毛发周期之期数、毛囊及毛干尺寸及毛发色素沉着程度。目测及使用低倍率(×8)解剖显微镜毛发长度。
此实施例及吾人国际申请案编号PCT/US 97/11033所述的实施例,显示平均约25日之C57BI/6及C3H老鼠的毛发生长周期,并记录毛囊及毛干发展之相同时序,而与所使用之除毛方法无关。
实施例2:豆浆及豆浆调配物之制备
一种制造豆浆之方法系将大豆浸于去离子水或纯水中历经数小时,于其完全水合之后进行研磨,添加少量水。(研磨方法使得豆浆被萃取)。收集之后,可过滤该豆浆以除去任何残留之豆皮部分。用于下述调配物之豆浆可为前述新鲜豆浆,或可自大豆粉及水制得。该大豆粉系自大豆研磨,亦可经冷冻干燥、喷干、或冷冻干燥,形成之豆浆可经或不经过滤。所制备之豆浆可含有约1至约90重量%之干燥大豆粉末。另一实施例系使用豆浆粉,由经冷冻干燥、喷干或冷冻干燥之豆浆制得,添加水或经过滤或均质化调理。亦可使用其他大豆萃取方法,以于下述调配物中产生该活性成分。例如,该活性成分可使用乙醇/水混合物而自经研磨之大豆萃取,如此保留该大豆之丝氨酸蛋白酶抑制活性,较佳系该蛋白质STI保持不变。
可用于本发明之豆制品可制自所有大豆类,而与其地理来源、曝晒、收割时间等因素无关。然而,以特定种类、地理来源或生长条件较佳。例如但不限于其大豆胰蛋白酶抑制剂(STI)含量或异黄酮含量丰富之大豆种类,或使豆类中之STI或异黄酮含量丰富之生长条件较佳。应已知可用于本发明组合物中之大豆产品具有明显气味,某些种类可让人接受,某些则较不受欢迎。若需要,则本发明组合物之气味可藉著使用由已知可减少气味之特定大豆种类衍生之大豆产品降低,包括但不限于脂肪氧合酶-2-缺乏型大豆及具有经修饰糖性质者等。于该调配物中降低氧浓度之方法亦可减少气味。亦可使用各种遮蔽剂或香料以掩盖该气味。
本发明组合物可另外包含表面活性剂、润湿剂、保湿剂、调理剂、香料、着色剂、防腐剂、抗氧化剂、脱色剂、除毛剂、抗老化剂、防晒剂、发泡剂、化妆佐剂、缓冲剂或其混合物。
本发明组合物可停留于皮肤上,历经足以产生改变之时间。例如,本发明组合物可应用于皮肤日常处理历经至少约四周,更佳系每日施加该组合物历经至少八周。
本发明另一种方法系为减少或预防假性须囊炎之方法。每日施加本发明组合物可减少或预防此种状况。本发明组合物亦可每日施加于该辅助区域以减缓毛发生长。此外,本发明组合物可每日施加于头皮,以造型或改善非裔型毛发之处理。
如吾人共待审美国专利申请案(代理文件编号JBP 430)所示,可使用数种以大豆为主之调配物以减少色素沉着。所有此等调配物亦可用于减缓毛发生长。下表2中列示部分特佳之豆浆调配物及含有异黄酮之豆浆调配物的实施例。用于本发明之异黄酮配制剂的实施例系为来自Ichimaru,Japan之黄固酮SE,含有约0.1%之纯异黄酮。所有此等调配物中,豆浆皆可使用适量之豆粉或豆浆粉及水置换。
表2:豆浆精油调配物:
大豆精油 | |||||||||
1 | 6 | 8 | 21 | 22 | 23 | 24 | 25 | 26 | |
豆浆 | 87.42% | 89.04% | 96.09% | 96.05% | 96.05% | 95.70% | 94.40% | 94.40% | 92.40% |
苯氧基乙醇 | 0.73% | ||||||||
苯氧基乙醇及对羟基苯甲酸酯 | 1.00% | 1.00% | 1.00% | 1.00% | 1.00% | 1.00% | 1.00% | 1.00% | |
甘油 | 2.50% | 2.50% | |||||||
环美沙酮(cyclomethicone) | 2.00% | ||||||||
铝淀粉琥珀酸辛酯 | 0.75% | ||||||||
蔗糖椰酸酯 | 1.00% | 1.00% | |||||||
PEG-6癸酸/辛酸三甘油酯 | 3.00% | 3.00% | |||||||
乙二胺四乙酸二钠 | 0.10% | 0.10% | 0.05% | 0.05% | 0.05% | 0.05% | |||
聚丙烯酰胺/月桂醇-7/C13-14异石蜡 | 2.50% | 2.75% | 2.90% | 2.90% | 2.90% | 3.20% | 3.50% | 3.50% | 3.50% |
抗坏血酸 | 0.01% | 1.00% | |||||||
丁基化的羟基甲苯 | 0.10% | 0.01% | 0.05% | 0.05% | 0.05% | 0.05% | 0.05% | 0.05% | |
聚山梨糖醇20 | 0.50% | ||||||||
乳铁蛋白 | 1.00% | 1.00% | 1.00% | ||||||
生育酚 | 1.00% | ||||||||
总量 | 100.00% | 100.00% | 100% | 100% | 100% | 100% | 100% | 100% | 100% |
27 | 28 | 29 | 30 | 31 | 32 | 33 | 34 | 35 | |
豆浆 | 90.70% | 94.70% | 85.7% | 90.7% | 93.70% | 94.70% | |||
苯氧基乙醇及对羟基苯甲酸酯 | 1.00% | 1.00% | 1.00% | 1.00% | 1.00% | 1.00% | 1.00% | 1.00% | 1.00% |
甘油 | 5.00% | ||||||||
乙二胺四乙酸二钠 | 0.05% | 0.05% | 0.05% | 0.05% | 0.05% | 0.05% | 0.05% | 0.05% | 0.05% |
聚丙烯酰胺/月桂醇-7/C13-14异石蜡 | 3.20% | 3.20% | 3.20% | 3.20% | 3.20% | 3.20% | 3.20% | 3.20% | 3.20% |
抗坏血酸 | |||||||||
丁基化的羟基甲苯 | 0.05% | 0.05% | 0.05% | 0.05% | 0.05% | 0.05% | 0.05% | 0.05% | 0.05% |
去离子水 | 90.70% | 90.70% | 85.70% | ||||||
道康宁200流体 | 1.00% | ||||||||
黄固酮SE | 10.0% | 5.00% | 2.00% | 1.00% | |||||
豆浆粉 | 5.00% | ||||||||
使用乙醇/水混合物之大豆萃取物 | 5% | 10% | |||||||
总量 | 100% | 100% | 100% | 100% | 100% | 100% | 100% | 100% | 100% |
实施例3:含有由大豆衍生之蛋白酶抑制剂之局部活性组合物的制备
购自Sigma-Aldrich Corporation之大豆胰蛋白酶抑制剂(STI)及Bowman-Birk抑制剂(BBI)混合于pH7.4之0.1M经磷酸盐缓冲之盐水(PBS,Gibco-BRL,Gaithersburg,MA)中,浓度为1%至0.001%。四份体积之形成之溶液与1份体积之(100毫克/毫升)脂质体媒质混合,该媒质系藉Niemiec等人所述之方法制备,以产生该局部活性组合物。非离子性脂质体配制剂,诸如Niemiec等人″非离子性脂质体组合物对于肽药物进入毛囊-皮脂腺单位之局部输送的影响:使用Hamster Ear模型进行体内研究″,12 Pharm.Res.1184-88(1995)(″Niemiec)(在此引用作为参考)中所揭示的,是本领域所熟知的,并且在吾人专利申请案(代理文件编号JBP 430)中有描述。GDL脂质体系如前述Niemiec等人所述般地制备,不同之处在于以下变化:该非离子性脂质体调配物含有甘油二月桂酸酯(Emulsynt GDL,ISP Van Dyk)/脂固醇(Croda)/聚氧化乙烯-10-硬脂醚(Brij76,ICI)/聚氧化乙烯-9-月桂醚,比例为37.5∶12.5∶33.3∶16.7。使用PBS或Hepes缓冲剂0.05M,pH7.4(Gibco-BRL ofGaithersburg,MD)作为制备该脂质体所使用之水相。
实施例4:豆浆延迟毛发生长并缩小毛囊及毛干尺寸并减少毛发色素沉着
C57B1/6老鼠系如实施例1所述般地诱导新毛发周期,每日使用豆浆处理。每日观察动物之毛发生长情况,于该毛发周期之重要时间点取下皮肤组织切片。使用豆浆处理之结果,延迟毛发生长,可见到其毛发变得稀薄,触感较为光滑。如同预期地,经处理之老鼠于该毛发周期之第7-8日未显示皮肤变黑,且与对照组动物不同地,于第11-12日未见到其毛干。经豆浆处理之老鼠的毛发周期平均延迟3-6日。图1系为老鼠毛皮之图型,显示于豆浆处理后之毛发外观、颜色、尺寸及厚度差异。
组织切片皮肤试样之组织检验确定此等观察结果。如Fontana-Mason(F&M)染色之图2所示,于毛发周期之第四日,未经处理之毛囊如同预期地完全发展,含有所有之细胞层及色素沉积。相反地,经豆浆处理之试样(于相同倍率下)显示较小而未完全发展之毛囊,而无色素沉积。
图3系显示两组使用F&M染色之组织剖面,其为低及高倍率。此等剖面系得自毛发周期之第七日。上层显示在与未经处理之对照组比较之下,经豆浆处理之皮肤具有较小、较短且较少色素沉着之毛囊。下层显示较高倍率之毛囊,进一步说明经豆浆处理之后的毛囊及毛干尺寸及色素沉着差异。
图4显示于毛发周期第18日时之经F&M染色皮肤的低倍率剖面图。于此倍率下,显然该豆浆处理使得毛囊尺寸缩小,而导致毛干长度及厚度缩小,并使得经处理毛囊之整体色素沉着减少。
图5系显示两倍率下之毛发周期第21日的皮肤剖面。上层说明对照动物系处于生长中期,此时毛囊退化。另一方面,经豆浆处理之毛囊已完成生长中期,因其已显示静止期,休止期。此情况显示经豆浆处理之后,不仅延迟毛发周期,亦使其提早终止。下层者于高倍率下说明该生长中期对照毛囊及较短、静止期(休止期)经豆浆处理毛囊。
实施例5:豆浆对于毛发生长、尺寸及色素沉着之影响可于C3H老鼠体内再现
为了证实豆浆对于毛发生长之影响并非C57B1/6老鼠所特有,吾人使用棕毛(Agouti)C3H老鼠重复实施例4所述之实验。此等实验之结果于目测及组织上皆相同。豆浆于C3H老鼠体内延迟毛发生长并缩小毛囊及毛干尺寸及色素沉着。
组织分析确定此等目测结果。如图6之上层所示,使用F&M染色,于该毛发周期之第七日,经豆浆处理之毛囊在与未经处理之对照组比较之下,较小而累积较少色素。图7之上层(F&M染色)显示较低倍率之相同皮肤剖面,说明经豆浆处理之后,较稀薄而较少色素沉着之毛囊。
图8系显示于毛发周期第21日时经F&M染色之皮肤剖面。如C57B1/6老鼠所示,经豆浆处理之后,该毛发周期提早终止。经豆浆处理之毛囊系处于休止期,而未经处理之对照毛囊仍处于生长中期。
实施例6:豆浆及由大豆衍生之丝氨酸蛋白酶抑制剂对于毛发生长、尺寸及色素沉着之影响
研究豆浆对于毛发生长之影响的机制之中,吾人试验由豆浆衍生之丝氨酸蛋白酶抑制剂STI及BBI的效果。吾人早期已证实此等蛋白质藉著影响PAR-2路径而于皮肤中诱发脱色(美国专利申请案,代理文件编号JBP 430)。
使用STI、BBI及豆浆,重复实施例4所述的实验。STI及BBI如实施例3所述般地用于PBS-脂质体媒质中。与豆浆相同地,该毛发周期中之视觉观察确定STI及BBI皆可延迟毛发生长,并缩小毛囊及毛干尺寸(参照图9之毛发图片)。使用高浓度之STI及BBI,对于毛发生长及色素沉着之影响极为实质。
组织分析确定此等发现。如图6所示,于毛发周期之第七日,1%STI及1%BBI于C3H老鼠体内缩小毛囊及毛干尺寸并减少毛干色素沉着。图7显示该毛发周期之同一日的低倍率剖面,说明相对于未经处理之对照组,使用豆浆、STI或BBI处理之毛囊及毛干较小并色素沉着较少。图10显示STI及BBI亦于C57B1/6老鼠体内具有相同效果,证明有较小且较少色素沉着之毛囊。综合起来,此实施例显示豆浆中所含之STI及BBI系为由大豆衍生之丝氨酸蛋白酶抑制剂,可延迟毛发生长,缩小毛囊及毛干尺寸,并减少毛发色素沉着。STI及BBI代表豆浆成分中影响毛发生长之一部分。
为了支持豆浆中之STI及BBI涉及前述毛发生长效应之假说,吾人试验豆浆之丝氨酸蛋白酶抑制活性。使用″Enzchek″进行检定,其系一种由Molecular Probeof Eugene,OR所制造之蛋白酶消化萤光试验系统。使用100单位之胰蛋白酶(购自Sigma chemicals,St.Louis MO),该试验系统产生约1100单位之萤光读数。此种反应系使用渐增浓度之STI抑制,如已知之胰蛋白酶抑制剂所预测。豆浆之连续稀释物于此检定中测试,发现其抑制胰蛋白酶活性。如图11所示,豆浆所具之胰蛋白酶抑制活性如同约0.2%纯度之STI。显示豆浆自身之毛发生长效应至少有一部分系来自STI及BBI。
实施例7:豆浆诱发酪氨酸酶及TRP-1蛋白质表达之改变
上述实施例所述经豆浆处理之皮肤样本的组织学分析显现毛囊内色素沉积减少。为进一步了解豆浆诱发之去色素作用的机制,吾等研究了酪氨酸酶(黑色素生成中关键的酶)及酶酪氨酸酶相关蛋白-1(TRP-1)(它稳定酪氨酸酶)。如上述处理C57B1/6及C3H鼠,于研究中全程收集样本以供蛋白质分析用。使用标准程序,如Juan S.Bonifacino等人编辑之细胞生物学第六章:电泳及免疫印迹中目前之程序(1999年著作权:John Wiley & Sons,Inc.),进行蛋白质萃取及Western印迹分析,该文献整体并于此以供参考用。如此研究之一范例示于图12。
等量之萃取自皮肤的蛋白质,用抗酪氨酸酶抗体“抗PEP1”以及抗-TRP-1抗体“抗PEP7”探测,其述于Jimeneze,M.,Kameyama,K.,Maloy,WL,Tomita Y.,及Hearing,V.哺乳动物酪氨酸酶:生合成、处理及藉由黑色素细胞刺激荷尔蒙之调节,Proc.Natl.Acad.Sci.USA(1988),85:3830-34,及Jimenez,M.,K.,Maloy,WL,及Hearing,V.酪氨酸酶克隆的专一性鉴别,J.Biol.Chem.(1989)264:3397-3403,其整体并于此以供参考用。
如图12所示,酪氨酸酶及TRP-1蛋白质之表达显著地由豆浆处理所影响。酪氨酸酶及TRP-1之量减少,及表达之持续期间缩短。此二因素影响整体毛发之色素沉着,其因黑色素生成之量减少及持续期间较短而减少。
实施例8:豆浆减少人类脸部毛发之长度及厚度
每天刮毛之具深色脸部毛发之男性个人在刮毛后立即以豆浆处理其脸右侧长达5周。在第3周前及更显著地在第4周前,经处理之该侧毛发可看出较淡且触摸感觉较柔顺。处理期间使用Hi-Scope在不同放大倍数下拍摄数位照片。此等照片清楚地证明处理侧中毛干尺寸及厚度之减小。如此照片之范例示于图13,证明四周处理后,毛干厚度及密度之差异。因脸部两侧均在同一时间刮毛且两侧照片系在同一时间拍摄,脸部毛发长度之差异显示在处理区域中较缓慢之生长速度。
图14显示豆浆处理四周后脸部毛发长度、厚度及全区之电脑化影像分析。所有影像以Image Pro Plus 3.0软体(Media Cybernetics,Silver Spring,MD)分析。数据以脸部各侧180根毛干之平均值与标准误差呈现(SigmaPlot(r)5.0,SPSS Science,Chicago,IL)。使用SigmaStat(r)2.0(SPSS Science)软体进行统计分析,证明豆浆处理后所有测量之参数中均有统计上显著的差异。
实施例9:豆浆减少人类脚部毛发长度及厚度
一个人膝盖下方脚中间部份之两对称区以蜡除毛。每天以豆浆处理一脚长达4周。视觉观察显示处理部位之毛发生长较慢。毛干之数目较未处理部位者少且较短及较稀疏,如图15所示。此等观察进一步确认豆浆对毛发生长之效果。实施例8及9一起确认豆浆对人类毛发生长之效果与除毛方法或被处理之身体部位无关。
实施例10:加入异黄酮之豆浆配方于降低毛发生长及色素沉着上较豆浆配方佳
使用表2所述之两个配方:豆浆为基质之配方豆浆精23及与豆浆精23相同但添加5%之0.1%异黄酮萃取物之豆浆精30重覆实施例4所述之实验。如图16所示,以大豆精23处理之鼠显现较少之毛发生长及较佳之毛发外观。此效果以使用用豆浆精30者较显著,证明添加异黄酮之豆浆配方在减少毛发生长上优于豆浆配方。图17显示在处理第15天时,经处理之鼠的组织皮肤断面。此等断面所考证之毛干清楚地证明豆浆精处理后毛干尺寸之减小,毛干内色素沉着量之减少及毛干柔顺度之增加。
实施例11:添加异黄酮之豆浆配方较异黄酮配方佳,其有效减少毛发生长及色素沉着
使用上述表2中之配方:含或不含增加浓度之异黄酮的豆浆精重覆实施例4所述之实验。此等豆精配方相当于类似配方,其中仅豆浆成份被水取代。制备此三组配方(豆精、异黄酮及含额外异黄酮之豆精)以测试黄酮可能足以对毛发生长产生所观察之效果的可能性。图18显示如实施例4所述三周局部处理后之C57B1/6鼠毛发。未经处理之对照组鼠及经安慰剂之鼠两者均具有长且较乱之毛发。如本案先前所叙述者,豆浆精23减少毛发生长且产生较佳之外观。含1、5及10%之含0.1%异黄酮萃取物的豆浆精配方对毛发生长的减少产生优异的效果。然而,含异黄酮而不含豆浆之配方证明较温和,但效果不如并用豆浆优异。此实施例证明含异黄酮之豆浆配方可减少毛发生长。此实施例进一步证明含异黄酮之豆浆配方较仅含异黄酮之配方更能减少毛发生长。
实施例12:豆精配方影响人类毛发生长
豆浆精23及30相较于安慰剂配方之功效及刺激性,在盲式安慰剂对照组和12周试验中被检视,其中有12个年龄29至45岁更年期前之女性试验者,其藉由研究调查者评估、试验者自我评估及Hi-scope影像分析检视。试验者签署被告知同意书并指示其研究程序及期望与要求当晚刮毛。第二天基线(baseline)探视时,分给各试验者二个试验化妆水(第1天):安慰剂化妆水及豆浆精23或30。化妆水随机给予右脚或左脚。在研究期间不使用其他化妆水于脚部下方,将试验化妆水及安慰剂用于各脚。试验者经指示每日施予试验化妆水早晚两次并被指示尝试不刮除脚部下方之毛发直至各周评估后。评估当天,调查者以视觉检查试验者脚部是否有任何临床刺激征状及比较脚部之毛发生长变稀疏之程度。在各评估时间点(第1、2、3及4周)由试验者完成自我评估问卷。此外,在各时间点使用MX-MACROZ镜头(Hirox)取得Hi-scope影像(各影像直径2.5公分,KH-2400R,Hirox)。
无一试验者系因任何产品相关之理由自研究退出。未在任一时间点在任一研究试验者看到刺激征状,亦未在研究期间任一时间点上于试验者所作自我评估中报告有任何刺激。
为计算包括短毛之所有毛发数目,在各时间点对各试验者进行2.5公分区域之计数。结果显示对于豆浆精30,在第5周前及对于豆浆精23,在第4周前脚部下方毛发数减少。虽然标准误差大,但经安慰剂处理之脚在研究全程中未显示平均脚毛发数有改变。生长率藉由试验者脚毛发之平均长度(其自在各试验部位使用Image Pro Plus分析对各试验者所作之hi-scope影像计算得到)除以自最后一次刮毛算起之时间长度(天)而计算得到。结果记录于表3,证明豆浆精23及30处理二者均较安慰剂产生较减缓之毛发生长速度。
表3:豆浆精或安慰剂处理后之平均脚毛发生长速度
周数 | 位置 | 豆浆精30 | 豆浆精23 | 安慰剂 |
0(基线) | 上方 | 0.211(±0.07) | 0.184(±0.07) | 0.243(±0.07) |
下方 | 0.248(±0.0) | 0.191(±0.06) | 0.235(±0.09) | |
1 | 上方 | 0.216(±0.05) | 0.153(±0.03) | 0.211(±0.06) |
下方 | 0.178(±0.08) | 0.213(±0.03) | 0.188(±0.07) | |
2 | 上方 | 0.232(±0.11) | 0.181(±0.04) | 0.221(±0.07) |
下方 | 0.236(±0.12) | 0.195(±0.07) | 0.217(±0.09) | |
3 | 上方 | 0.241(±0.06) | 0.185(±0.09) | 0.285(±0.13) |
下方 | 0.213(±0.10) | 0.147(±0.04) | 0.253(±0.12) | |
4 | 上方 | 0.234(±0.07) | 0.209(±0.11) | 0.211(±0.09) |
下方 | 0.194(±0.05) | 0.208(±0.07) | 0.220(±0.04) |
自试验者自我评估问卷之结果显示,试验者感觉试验化妆水相较于安慰剂化妆水减少毛发之生长及软化脚毛发之触觉。试验者感觉毛发较不粗糙且短毛较少。大部份之试验者相信试验化妆水减少脚毛发之生长或改变毛发之纹理以致感觉较柔顺及较不粗糙。Hi Scope分析进一步证明以豆浆精23或30处理后毛发似乎以相同方向再生长且形状、纹理及外观更均匀。相反地,在经安慰剂处理之脚上再生长之毛发以不同方向、不同长度、不同生长角度及厚度生长。
此实施例清楚地证明豆浆配方于延缓及减少毛发生长并使毛发较柔软、较不粗糙及更易处理与定向生长之效果。
Claims (7)
1.一种改变哺乳动物毛发外观、毛发生长、毛发色素沉着及毛囊与毛干尺寸的局部活性组合物,其特征在于,该组合物含有豆浆和基于豆浆体积0.1-15%体积/体积的异黄酮;所述豆浆是通过浸泡和研磨大豆,加入水并过滤除去豆皮的剩余部分制备的。
2.如权利要求1所述的组合物,其中所述豆浆含量为10-99%体积/体积,以局部活性组合物的总体积为基准。
3.如权利要求1所述的组合物,其特征在于,其中所述豆浆含量为50-99%体积/体积,以局部活性组合物的总体积为基准。
4.如权利要求1所述的组合物,其中该局部活性组合物进一步包含制药或化妆品可接受的赋形剂。
5.如权利要求1所述的组合物,其特征在于,所述异黄酮含量为基于豆浆体积0.1-10%体积/体积的异黄酮。
6.如权利要求5所述的组合物,其中该局部活性组合物进一步包含制药或化妆品可接受的赋形剂。
7.权利要求1所述的局部活性组合物的用途,其特征在于,该用途是用于制备使哺乳动物毛发生长迟缓、毛囊及毛干尺寸缩小及毛发色素沉着减少的药物。
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Also Published As
Publication number | Publication date |
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CA2314562C (en) | 2011-08-23 |
CN1316272A (zh) | 2001-10-10 |
DE60044697D1 (de) | 2010-09-02 |
HK1032540A1 (en) | 2001-07-27 |
US7985404B1 (en) | 2011-07-26 |
EP1074240A3 (en) | 2003-01-29 |
US20040126353A1 (en) | 2004-07-01 |
US20030064048A1 (en) | 2003-04-03 |
ES2347316T3 (es) | 2010-10-28 |
ATE474551T1 (de) | 2010-08-15 |
CA2314562A1 (en) | 2001-01-27 |
JP2001072555A (ja) | 2001-03-21 |
EP1074240B1 (en) | 2010-07-21 |
EP1074240A2 (en) | 2001-02-07 |
KR20010015444A (ko) | 2001-02-26 |
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