TWI830748B - 防脫髮或毛髮生長促進用組合物 - Google Patents
防脫髮或毛髮生長促進用組合物 Download PDFInfo
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- TWI830748B TWI830748B TW108124176A TW108124176A TWI830748B TW I830748 B TWI830748 B TW I830748B TW 108124176 A TW108124176 A TW 108124176A TW 108124176 A TW108124176 A TW 108124176A TW I830748 B TWI830748 B TW I830748B
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- hair
- composition
- hair growth
- promoting
- hair loss
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Abstract
本發明涉及防脫髮或生髮促進用組合物,其包含誘導生髮或促進毛髮生長的成分,用於促進毛髮生長。本發明的防脫髮及毛髮生長促進用組合物縮短在毛髮的生長週期中從休止期轉移到生長期的週期,從而促進毛髮的生長,延緩向退行期的轉換,防脫髮及毛髮生長促進效果優異。另外,本發明的組合物對人體安全且無副作用,並具有優異的促進毛乳頭細胞增殖,增強毛髮彈性的效果。
Description
本發明涉及用於促進毛髮生長的組合物,涉及包含誘導生髮或促進毛髮生長的成分的用於促進毛髮生長的防脫髮及生髮促進用組合物。
在隨著時代的改變而對美容的關注越來越高的過程中,毛髮佔據著重要的一部分。毛髮是在皮膚表面生出的細且被角質化的結構。其具有對外部衝擊的緩衝作用,且保護人體不受直射光線、寒冷、摩擦、危險等外部刺激,執行將對身體有害的砷、汞、鋅等重金屬排出到體外的功能,近來還被強調裝飾上的美容效果。但是,因飲食習慣的改變或內外壓力的增加等各種原因,受到脫髮困擾的人越來越多。
人的毛髮約為10萬-15萬根左右,各個毛髮具備彼此不同的生長週期。毛髮生長週期構成為3個階段,被劃分成毛髮生長最為活躍的生長期(anagen stage)、毛髮開始退化的退化期(catagen stage)及停止毛髮的生長或進入休息階段的休止期(telogen stage)。
在對這樣的毛髮的生長週期進行調節時,起到重要的作用是包含構成毛囊的毛乳頭細胞及毛囊幹細胞的外毛根鞘細胞(outer root sheath cell)的活性和在這些細胞中生出的多種細胞因數(cytokine)及生長因數。例如DKK-1(Dickkopf-1)對阻礙和殺滅毛囊生長起到重要作用(J Invest. Dermatol,2008:128(2))。另一方面,為了週期性的毛髮生長,需要形成毛囊的再生及幹細胞的啟動,而這是通過Wnt/β-連環蛋白的生成及啟動而實現的(Nature2007:447(7142))。
在一般情況下,脫髮症是指如下情況:在這樣的週期中生長期的毛髮比率變小,退行期或休止期的毛髮變多,由此毛髮脫落的數量異常增多。相對於正常人的生長期狀態的毛髮多的情況,患有脫髮症(Alopecia)的人是休止期狀態的毛髮多,因此出現肉眼可見的脫髮現象。出現脫髮症的人群的特徵在於毛髮的小型化。越出現脫髮,生長期的期限越短,促進向退行期及休止期的轉移,之後毛乳頭的體積變小,毛囊逐漸被小型化。因此,為了治療脫髮,重要的是將休止期狀態的毛囊快速恢復為生長期,將被縮短的生長期延長。
作為脫髮的原因,提出了雄性荷爾蒙作用過度的說法、皮脂分泌過度的說法、血液迴圈不良的說法、由過氧化物、細菌等引起的頭皮功能下降的說法、遺傳性因素、老化、壓力等說法,但目前為止脫髮的確切原因尚未被揭開。
在毛囊的生長和脫落中,雄性荷爾蒙起到非常重要的作用,在通過雄性荷爾蒙而在毛囊中被調節的因數中,TGF-β對由雄性荷爾蒙導致的毛囊生長的阻礙和毛囊週期中的向退行期的誘導起到重要的作用(Tsuji Y,Denda S,Soma T,Raftery L,Momoi T,Hibino T,A potential suppressor of TGF-beta delays catagen progression in hair follicles,J Investig Dermatol Symp Proc,2003:8(1):65-68)。即,根據雄性荷爾蒙而在毛囊的毛乳頭細胞中表達而引起毛囊的基質細胞和外根鞘角質細胞的細胞凋亡,將生長期的毛囊誘導至退行期,從而當TGF-β的數量增加時,誘導毛母質細胞(hair matrix cell)和外根鞘細胞的細胞凋亡,由此休止期的毛囊增加且生長期的毛囊轉換為退行期的毛囊而容易脫落,從而生成又細又短的毛髮(李英珠(이영주)、李珠英(이주영),男性的雄激素性脫髮者的臨床診斷和管理,韓國美容協會雜誌,2007:13(2):799-810)。因此,公知為TGF-β抑制毛囊生長。
七葉皂素(escin)是大量包含在七葉樹提取物中的皂素系列的物質,其主要被用作抗炎作用、血管收縮劑或血管保護劑。而在本發明之前,從未公開七葉皂素的改善脫髮並促進生髮的效果。
目前為止所研發的脫髮症的治療或預防用製劑有將用於促進血液迴圈,強化髮根功能,保濕頭皮及抑制雄性荷爾蒙的女性荷爾蒙為主成分的製劑或含有米諾地爾(minoxidil)、非那雄胺(Finasteride)、黏多醣生長素(trichosaccharide)的製劑等。代表性的塗布用脫髮治療劑即米諾地爾(2,4-diamino-6-piperidinopyrimidine-3-oxide)增加流向頭皮的血流量,口服用脫髮治療劑即非那雄胺起到抑制5α-reductase的活性而減少活性型雄性荷爾蒙即DHT的生成的功能。但是,上述製劑因副作用或使用上的注意事項而在使用上存在諸多限制。
[發明要解決的課題]
本發明要解決的課題在於提供一種防脫髮或毛髮生長促進用組合物。
另外,本發明要解決的課題在於提供包含上述防脫髮或毛髮生長促進用組合物的頭髮或頭皮用產品。
[用於解決課題的手段]
本發明人為了研製出改善以往的脫髮治療劑的副作用及使用上的注意事項等問題並克服了防脫髮/養髮劑的生髮促進效果微弱等的缺點且對人體安全且能夠有效地促進毛髮生長及生髮的組合物而不懈努力,其結果發現七葉皂素對防脫髮及促進毛髮生長具有顯著的優異效果,從而完成了本發明。
具體地,本發明的發明人確認到七葉皂素在毛乳頭細胞中促進Wnt/β-連環蛋白信號傳送系統的活性,促進毛乳頭細胞的增殖,增加β-連環蛋白的表達量。即,通過體外(in-vitro)實驗而確認了七葉皂素的如下的重要作用:放大毛乳頭細胞的Wnt/β-連環蛋白信號傳送,從而將毛髮的生理週期從退行期轉移到活性期。另外,本發明的發明人確認了在將包含七葉皂素的脫髮治療用組合物使用於脫髮症患者時,對改善毛髮的粗細、茂密程度及彈性具有顯著的優異效果。另外,與市場上銷售的產品即米諾地爾比較而進行實驗的結果,確認了本發明的包含七葉皂素的脫髮治療用組合物與米諾地爾相比,具備類似或更加優異的防脫髮及生髮效果。
本發明提供作為有效成分而包含七葉皂素的防脫髮或毛髮生長促進用組合物。
在一個實施例中,作為本發明的防脫髮或毛髮生長促進用組合物的有效成分而包含的七葉皂素的含量相對於組合物總重量為0.001重量%至10重量%,優選為0.01重量%至5重量%,更優選為0.1重量%至2重量%。在本發明中,上述七葉皂素相對於組合物總重量為0.001重量%至10重量%的情況下,體外(in-vitro)或臨床上顯示出對防脫髮及促進毛髮生長具有優異的效果。在上述七葉皂素低於0.001重量%的情況下,毛髮生長的誘導促進效果微弱,在超過10重量%的情況下,可能出現阻礙劑型的安全性的問題和皮膚刺激。更優選為,上述七葉皂素的含量相對於組合物的總重量為0.5重量%至2重量%,最優選為1至2重量%。在本發明中,通過實驗而確認了七葉皂素在上述含量範圍內時,促進生髮及防脫髮效果顯著。
本發明的七葉皂素可作為防脫髮劑或毛髮生長促進劑而包含在上述組合物或劑型。
另外,本發明的一個觀點提供一種作為有效成分包含選自由土大黃苷(Rhaponticin)、大黃酸(Rhein)、大黃酚(Chrysophanol)及大黃素甲醚-8-O-d-吡喃葡萄糖苷(Physicon-8-O-d-glucopyranoside)構成的組的1種以上的化合物或其鹽、水合物或溶劑化物的防脫髮或毛髮生長促進用組合物。
土大黃苷作為二苯基葡萄糖苷化合物,治療糖尿病及阿茲海默症潛力巨大,可表示為下述化學式。
大黃酸(Rhein)作為又被稱作山扁豆酸的蒽醌組的化合物,具備抗菌及抗生特性,可表示為下述化學式。
大黃酚(Chrysophanol)作為從菌類分離出的天然的蒽醌組的化合物,可表示為下述化學式。
大黃素甲醚-8-O-d-吡喃葡萄糖苷(Physicon-8-O-d-glucopyranoside)作為天然蒽醌衍生物,可表示為下述化學式。
在一個實施例中,本發明的有效成分含有選自上述化合物的1種以上化合物,優選為含有2種以上化合物,在將2種以上化合物組合使用的情況下,對防脫髮及促進毛髮生長顯示出最佳效果。
在一個實施例中,本發明的有效成分含有2種以上的化合物,上述2種以上的化合物可以是選自(i)大黃酚及以及(ii)土大黃苷、大黃酸及大黃素甲醚-8-O-d-吡喃葡萄糖苷的任意一個組合。
在另一個實施例中,上述2種以上的化合物可以是選自(i)土大黃苷及(ii)蒽醌衍生化合物(大黃酸、大黃酚及大黃素甲醚-8-O-d-吡喃葡萄糖苷)的任意一個組合。
在又一個實施例中,上述2種以上的化合物是土大黃苷、大黃酸、大黃酚及大黃素甲醚-8-O-d-吡喃葡萄糖苷這4種化合物的組合。
另外,在本發明的組合物中,用作有效成分的不僅是上述化合物本身,還可以是其鹽、水合物或溶劑化物。
上述鹽可利用鹽酸鹽、氫溴酸鹽或氫碘酸鹽這樣的無機酸和如下的有機酸而形成:醋酸鹽、己二酸鹽、藻朊酸鹽、天冬氨酸鹽、甲苯酸鹽、苯磺酸鹽、p-甲苯磺酸鹽、硫酸氫鹽、氨基磺酸鹽、硫酸鹽、萘酸鹽、丁酸鹽、檸檬酸鹽、樟腦酸鹽、樟腦磺酸鹽、環戊丙酸睾酮、雙葡萄糖酸鹽、十二烷基硫酸鹽、乙烷磺酸鹽、富馬酸鹽、葡庚糖酸鹽、甘油磷酸鹽、半硫酸鹽、庚酸鹽、己酸鹽、2-羥基乙烷磺酸鹽、乳酸鹽、馬來酸鹽、甲磺酸鹽、2-萘磺酸鹽、煙酸鹽、草酸鹽、甲苯磺酸鹽或十一烷酸鹽。
水合物是指,在上述化合物中添加了水或水進入到分子的結構物質的物質,溶劑化物是指,溶質的分子或離子與溶劑的分子或離子之間產生的高階化合物。上述水合物或溶劑化物可利用上述的酸而製得。
本發明的組合物包括上述1種以上的化合物或其鹽、溶劑化物或水合物的有效量,因此可實現促進毛乳頭細胞的增殖和/或增強毛髮的彈性的效果。
上述有效量是指,足以預防、改善及治療脫髮的量。上述有效量可以根據疾病的症狀程度、患者的年齡、體重、健康狀態、性別、投給途徑及治療期限等而適當改變。
具體地,在本發明的組合物中,上述有效量可以為0.00001重量%至50重量%。具體地,本發明包含0.00001重量%至50重量%的土大黃苷,優選包含0.0001重量%至0.1重量%的土大黃苷,或包含0.00001重量%至50重量%的大黃酸,優選包含0.0001重量%至0.1重量%的大黃酸,或包含0.000001重量%至50重量%的大黃酚,優選包含0.0001重量%至0.01重量%的大黃酚,或包含0.00001重量%至50重量%的大黃素甲醚-8-O-d-吡喃葡萄糖苷,優選包含0.0001重量%至0.1重量%的大黃素甲醚-8-O-d-吡喃葡萄糖苷。
這是因為,當土大黃苷(Rhaponticin)、大黃酸(Rhein)、大黃酚(Chrysophanol)及大黃素甲醚-8-O-d-吡喃葡萄糖苷(Physicon-8-O-d-glucopyranoside)的組成物質低於0.00001重量%時,毛髮的生長誘導促進效果微弱,在超過50重量%的濃度下,劑型的安全性下降而不優選。
在一個實施例中,可將本發明的防脫髮或毛髮生長促進用組合物用作醫藥組合物、醫藥外品用組合物或化妝料組合物。可將本發明的組合物劑型化為通常可適用於皮膚的任何形態,但優選劑型化為皮膚外用劑的形態。可將本發明的組合物製造成例如液態、乳狀、膏狀或固體狀等適用於皮膚的劑型。
作為本發明的一個形態,上述組合物可以是用於預防或治療脫髮的醫藥組合物。在該情況下,防脫髮或毛髮生長促進用組合物包含上述的1種以上的化合物、其鹽、水合物或溶劑化物的有效量及一個以上的藥學上允許的載體、賦形劑或稀釋劑。
上述說明中「藥學上允許」是指,在生理學上允許且在投入到人體時,通常不引起胃腸功能紊亂、眩暈這樣的過敏反應或與此類似的反應的組合物。
作為上述載體、賦形劑及稀釋劑的例子,可例舉乳糖、葡萄糖、蔗糖、山梨糖醇、甘露醇、木糖醇、赤蘚糖醇、麥芽糖醇、澱粉、***樹膠、藻蛋白酸鹽、明膠、磷酸鈣、矽酸鈣、纖維素、甲基纖維素、聚乙烯吡咯烷酮、水、羥苯甲酯、羥苯丙酯、滑石、硬脂酸鎂及礦物油。另外,還可進一步包含填充劑、抗凝劑、潤滑劑、濕潤劑、香料、乳濁劑及防腐劑等。
作為本發明的另一形態,上述組合物可以是醫藥外品用組合物。在該情況下,可將本發明的有效成分直接添加或與其他醫藥外品成分一起使用,並且可通過通常的方法而適當地使用。關於這樣的其他成分,可根據醫藥外品的劑型或使用目的等而任意地選擇來進行配合。例如,可包含增稠劑、穩定劑、溶解劑、維生素、顏料及香料這樣的通常的輔助劑及載體等。
作為本發明的又一形態,上述組合物可以是用於防止、改善脫髮或促進毛髮生長的化妝料組合物。
在本發明的劑型為液態的情況下,作為載體成分,可使用溶劑、溶解劑或乳濁劑等,例如可使用水、乙醇、異丙醇、乙基碳酸酯、乙基醋酸鹽、苄醇、甲苯酸鹽苄酯、丙二醇、1,3-丁基乙二醇油、甘油脂肪族酯、聚乙二醇或山梨糖醇的脂肪酸酯等。
在本發明的劑型為膏、霜或膠的情況下,作為載體成分可使用動物性油、植物性油、蠟、石蠟、澱粉、西黃蓍膠、纖維素衍生物、聚乙二醇、矽、皂黏土、二氧化矽、滑石或氧化鋅等。
在本發明的劑型為粉末或噴霧的情況下,作為載體成分可利用乳糖、滑石、二氧化矽、氫氧化鋁、矽酸鈣或聚醯胺粉末等。特別地,在噴霧劑型的情況下,可進一步包含氯氟烴、丙烷/丁烷或二甲醚等這樣的推進劑。
關於包含在本發明的組合物中的成分,除了上述有效成分之外,還可包含適用於皮膚的外用劑中通常利用的成分。例如可追加包含選自由水、表面活性劑、保濕劑、低級醇、螯合劑、殺菌劑、防氧化劑、防腐劑、色素、香料、增稠劑、研磨劑、甜味劑、pH調節劑、結合劑、發泡劑、溶解劑、維生素及顏料等構成的組的一個以上的添加劑。
另外,本發明提供包含上述防脫髮或毛髮生長促進用組合物的頭髮或頭皮用產品。上述頭髮或頭皮用產品可以是選自由生髮劑、頭皮護理劑、頭皮清潔劑、頭皮按摩劑、頭皮保養劑、清洗劑、洗髮水、滋潤劑、護髮素、髮乳、膠、髮膜、霜、精華、粉末、噴霧、油、皂、軟膏、頭髮造型劑、染髮劑及燙劑等構成的組的任意一個,但不限於此。
在本發明的一個實施例中,將上述防脫髮或毛髮生長促進用組合物製造成頭髮滋潤劑或髮乳的劑型(實施例1至13)。
優選為,通過直接塗布或噴灑到皮膚等經皮投給的方法來使用本發明的毛髮生長促進用組合物。
本發明中使用的用語「投給」是指,通過某種適當的方法來導入本發明的組合物。關於本發明的組合物的投給途徑,只要能夠達到目標群組織,可通過任意的一般路徑而投給。優選為,可經皮投給,其中最優選為局部塗布。本發明的組合物的適用次數可根據處方、需要或期望而決定。
關於本發明的組合物的使用量,可根據年齡、病變的程度等個人差異或劑型而適當調節,通常優選以1天1次至數次地將適當量塗布到頭皮的方式使用一周至數月。在本發明的實施例中,將脫髮治療用組合物1以1周適用5次的方式使用了6個月,其結果確認到優異的毛髮生長促進效果(實驗例4及8)。
[發明效果]
本發明的防脫髮及毛髮生長促進用組合物縮短毛髮的生長週期中從休止期轉移到生長期的週期,從而促進毛髮的生長,延緩向退行期的轉換,由此實現優異的防脫髮及促進毛髮生長的效果。
另外,本發明的組合物對人體安全無副作用且實現優異的防脫髮及促進毛髮生長的效果。另外,本發明的防脫髮或毛髮生長促進用組合物對促進毛乳頭細胞的增殖,增強毛髮彈性方面起到非常顯著的作用。
下面,為了幫助理解本發明,通過實施例等而進行詳細說明。但是,本發明的實施例可變形為各種其他形態,本發明的範圍不應僅限於下述實施例而解釋。本發明的實施例是為了向所屬技術領域中具有通常知識者更完整地說明本發明而提供的。
製造例1:脫髮治療/生髮用組合物1(頭髮滋潤劑)
按照下述表1所示的處方,通過通常的方法製造了包含七葉皂素的實施例及比較例的頭髮滋潤劑。
[表1]
製造例2:脫髮治療/生髮用組合物2(髮乳)
按照下述表2所述的處方,通過通常的方法製造了包含七葉皂素的髮乳(實施例7)。
[表2]
實驗例1:毛乳頭細胞活性信號(Wnt/β-連環蛋白)調節效果
在一般情況下,在毛乳頭中被啟動的wnt/β-連環蛋白信號在毛髮的生長週期中從退行期過度到活性期時,即從毛髮開始生出到發生毛髮生長的活性期內發生。另外,在休止期及退行期,wnt/β-連環蛋白信號變弱或消失,由此出現毛囊退化而毛髮脫落的現象。在本實驗例中確認了七葉皂素對wnt/β-連環蛋白信號放大起到多大的作用。
作為放大Wnt/β-連環蛋白信號的陽性對照群使用了wnt3a蛋白質,並作為陰性對照群使用了DMSO。在培養96-孔板中,將約3x104
的Wnt報告HEK293A細胞播種(seeding)到各個孔中,然後對七葉皂素分別進行了5 µg/ml、10 µg/ml、20 µg/ml的濃度處理。之後利用Promega公司的螢光素酶分析試劑盒(Luciferase assay kit)(E1960),按照製造公司的實驗方法而進行了報告分析。利用Luminometer Victor(PerkinElmer,Waltham,Massachusetts,USA)檢測了Wnt/β-連環蛋白啟動子的活性度,分析結果如圖1所示。
從實驗結果來看,七葉皂素將相對無處理群即陰性對照群的Wnt/β-連環蛋白信號放大了11倍左右。包含七葉皂素的實驗群未顯示出依賴於濃度的狀況,但在20µg/ml的濃度附近反覆確認到了可靠的效果。特別地,七葉皂素比結合到Wnt/β-連環蛋白的直接的收納體的陽性對照群即wnt3a蛋白質顯示出了優異的效果。因此,確認到七葉皂素大大促進毛囊幹細胞的毛髮生長促進信號即Wnt/β-連環蛋白信號傳送系統的活性,從而具備顯著的生髮促進效果。
實驗例2:毛乳頭細胞增殖促進效果
從PromoCell公司購買了源自人體的毛乳頭細胞(dermal papilla cells,DPCs)。使用由PromoCell公司推薦的毛囊真皮乳頭細胞生長培養基(Follicle Dermal Papilla Cell Growth Medium)及補充劑混合物(supplement mix),在37℃及5%的CO2
條件下培養了上述DPCs。將所培養的3,000cells/well的DPCs分注到96-孔板,然後在0.1%血清條件下培養了24小時。之後,將在所培養的細胞的無血清DMEM中按照1:1000進行稀釋的DMSO(vehicle)作為對照群,對七葉皂素分別按照0.2 µg/ml、0.5 µg/ml、1 µg/ml、2 µg/ml及5 µg/ml進行1天的處理並進行了孵化。孵化之後利用CCK方法對細胞的增殖程度進行了評價。將CCK-8在培養液中按照1:10進行處理並進行了1小時的孵化。在1小時之後,檢測各個孔的吸光度(absorbance)為450nm。所有實驗反覆進行了3次,並計算了吸光度值的平均值。關於上述結果,在將對照群設為100的情況下,表現為處理群對其的百分比。實驗結果,在處理了七葉皂素的情況下,顯示出DPCs增殖效果,確認到毛乳頭細胞增殖效果優異。
[表3]
實驗例3:毛乳頭細胞β-連環蛋白的表達增加
在Wnt信號被抑制的休止期及退行期的毛乳頭細胞中β-連環蛋白被GSK3-β而分解。但在Wnt信號被放大的活性期毛乳頭細胞中GSK3-β的活性被抑制,因此β-連環蛋白不被分解,因此β-連環蛋白的表達量增加。將人體毛乳頭細胞分注到約500,000cell/well的6個孔板,然後對七葉皂素分別按照0.2 µg/ml、0.5 µg/ml、1 µg/ml、2 µg/ml及5 µg/ml進行1天的處理並進行了孵化。在孵化之後利用M-PER溶解緩衝液而溶解細胞,將相同量的細胞溶解物加入到SDS-PAGE膠。將加入到SDS-PAGE膠的細胞溶解物移送到硝化纖維素膜,然後將抗-β-連環蛋白抗體黏貼到硝化纖維素膜,檢測了蛋白質。將GAPDH用作校正蛋白質量的控制器(control)。確認了通過七葉皂素增加的β-連環蛋白的量,並將其結果顯示於圖2。
實驗結果確認到與無處理群相比,在處理了七葉皂素的組中β-連環蛋白的量得到很顯著的增加,這表示毛乳頭細胞的Wnt/β-連環蛋白信號得到增強。
實驗例4:脫髮治療/生髮用組合物1(頭髮滋潤劑)的生髮效果確認
以毛髮的數量比正常情況明顯少或具有脫髮症狀的毛髮不好的共計105名的男女為對象,對本發明的脫髮治療/生髮用組合物1(頭髮滋潤劑)的生髮效果進行了實驗。將上述共計105名的男女按照15名分為7組而使用了比較例1及實施例1至6,一周5次地對毛髮及頭皮使用了6個月。在使用之後,關於毛髮的粗細、茂密程度、彈性及整體評價專案,分別分成5階段而評價了改善程度或損傷程度(改善程度:+5,+4,+3,+2,+1,無改善:0,損傷程度:-1,-2,-3,-4,-5)。上述評價是在開始使用起6個月之後根據使用者的問診結果的平均值而進行判定的,其結果如表4所示。
[表4]
從上述結果可確認,在包含七葉皂素的實施例1至6中顯示出較高的防脫髮及毛髮生長效果。由此,證明了作為有效成分包含七葉皂素的本發明的防脫髮及毛髮生長促進用組合物可非常有效地使用於脫髮治療。
實驗例5:脫髮治療/生髮用組合物1(頭髮滋潤劑)的生髮效果確認
以毛髮的數量比正常情況明顯少或具有脫髮症狀的毛髮不好的共計105名的男女為對象對本發明的脫髮治療/生髮用組合物1(頭髮滋潤劑)的生髮效果進行了實驗。將上述共計105名的男女按照15名分為7組而使用了比較例1及實施例1至6,1周5次地對毛髮及頭皮使用了6個月。在使用之後通過臨床照片拍攝進行了研究者評價,並通過頭皮毛髮圖像分析而進行了評價。關於通過臨床照片進行的研究者評價,在塗布2個月、4個月、6個月之後對比較例和實驗例進行了「良好(good)」、「微好(slight)」、「無變化(no change)」這3個專案的評價(Good:好轉50%~75%,Slight:好轉25%~50%,no change:無變化)。在塗布4個月、6個月之後通過頭皮毛髮圖像分析而對比較例及實施例檢測每單位面積的毛髮的數量和平均毛髮的厚度並進行了評價,其結果如表5所示。
[表5]
從上述結果可確認,在包含七葉皂素的實施例1至6中顯示出較高的防脫髮及毛髮生長效果。由此,證明了作為有效成分包含七葉皂素的本發明的防脫髮及毛髮生長促進用組合物可非常有效地使用於脫髮治療。
上述頭髮滋潤劑或髮乳這樣的劑型例僅為本發明的毛髮生長促進用組合物的例示,所屬技術領域中具有通常知識者顯然應該明白本發明的組合物的範圍不限於這些劑型。
製造例3:脫髮治療用組合物3(頭髮滋潤劑)
按照下述表6所述的處方,通過通常的方法利用土大黃苷(Rhaponticin)、大黃酸(Rhein)、大黃酚(Chrysophanol)及大黃素甲醚-8-O-d-吡喃葡萄糖苷(Physicon-8-O-d-glucopyranoside)中的1種以上而製造了頭髮滋潤劑。
[表6]
製造例4:脫髮治療用組合物4(髮乳)
按照下述表7所示的處方,通過通常的方法利用土大黃苷、大黃酸、大黃酚及大黃素甲醚-8-O-d-吡喃葡萄糖苷中的1種以上而製造了髮乳(實施例13)。
[表7]
實驗例6:毛乳頭細胞增殖促進效果
從PromoCell公司購買了源自人體的毛乳頭細胞(dermal papilla cells,DPCs)。在包含5%的胎牛血清(FBS;Gibco,NY,USA)、100 unit/mL的青黴素及100µg/ml的鏈黴素的DMEM(Hyclone Inc,UT,USA)中在37℃及5%的CO2
的條件下培養了上述DPCs。將培養的3,000cells/well的DPCs分注到96-孔板,然後在0.1%血清條件下培養了24小時。之後,將在所培養的細胞的無血清DMEM中按照1:1000稀釋的DMSO(vehicle)作為對照群,將2 µg/ml的米諾地爾作為陽性對照群,對將各個10 µg/ml的土大黃苷、大黃酸、大黃酚及大黃素甲醚-8-O-d-吡喃葡萄糖苷及表8所示的單一物質組合而成的試樣進行1天的處理並進行了孵化。在孵化之後利用CCK方法對細胞的增殖程度進行了評價。在培養液中將CCK-8按照1:10進行處理而孵化了1小時。在1小時之後,檢測各個孔的吸光度(absorbance)為450nm。將所有實驗分別反覆進行3次,計算了吸光度值的平均值。關於上述結果,在將對照群設為100的情況下,表現為處理群對其的百分比。
其結果,在對土大黃苷、大黃酸、大黃酚、大黃素甲醚-8-O-d-吡喃葡萄糖苷進行處理的情況下,關於DPCs增殖促進效果表示出了優異的毛乳頭細胞增殖效果(表8)。
[表8]
實驗例7:毛乳頭細胞活性信號調節效果
在一般情況下,在毛乳頭中分泌的VEGF、IGF-1、FGF等誘導毛髮的生長期,Versican被公知為毛髮生長期的標誌,DKK-1被公知為毛髮的退行期的標誌。因此,對本發明的土大黃苷、大黃酸、大黃酚及大黃素甲醚-8-O-d-吡喃葡萄糖苷的物質在毛乳頭細胞中對生長期促進信號和退行期誘導信號產生的影響進行了調查。
在培養6-孔板中,將毛乳頭細胞1x105
分別播種(seeding)到各個孔之後,對將各個10 µg/ml的本發明的土大黃苷、大黃酸、大黃酚及大黃素甲醚-8-O-d-吡喃葡萄糖苷的物質及表10所示的單一物質組合而成的試樣進行處理,然後利用即時PCR方法,並利用下述表9的Taqmanⓡ
探針(Thermo Fisher,Massachusetts,USA)而對VEGF,versican(多功能蛋白聚糖)及DKK-1的表達量進行了調查,其結果如表10所示。
[表9]
其結果,土大黃苷、大黃酸、大黃酚及大黃素甲醚-8-O-d-吡喃葡萄糖苷的物質中增加了毛髮生長期誘導生長因數即VEGF和生長期標誌即Versican的表達,降低了毛髮的退行期誘導因數DKK-1的表達(表10)。
[表10]
如以上所記載,確認到土大黃苷、大黃酸、大黃酚及大黃素甲醚-8-O-d-吡喃葡萄糖苷的物質具備優異的毛髮生長誘導及脫髮抑制效果,並將本發明的防脫髮及毛髮生長促進用組合物製造成了各種劑型。下述記載中對其劑型例進行了具體說明。但是,這些劑型例僅為本發明的毛髮生長促進用組合物的例示,所屬技術領域中具有通常知識者顯然應該明白本發明的組合物的範圍不限於這些劑型。
實驗例8:脫髮治療用組合物3(頭髮滋潤劑)的生髮效果確認實驗
關於上述製造例3中製造的本發明的脫髮治療用組合物3(頭髮滋潤劑),以毛髮的數量比正常情況明顯少或出現脫髮症狀的毛髮不好的共計40名的男女為物件,按照10名分為4組,將比較例2、3及實施例8至12,1周5次地對毛髮及頭皮使用了6個月。在使用6個月之後,關於毛髮的粗細、茂密程度、彈性及整體評價專案,通過對使用者進行問診而得到的平均值來判定了改善程度(變得非常好;+3,變好;+2,稍微變好;+1,無變化;0,稍微變壞;-1,變壞;-2,變得非常不好;-3),其結果如表11所示。
[表11]
從結果可知,在適用了土大黃苷、大黃酸、大黃酚及大黃素甲醚-8-O-d-吡喃葡萄糖苷的物質的實施例中顯示出了較高的防脫髮及毛髮生長效果,由此證明了本發明能夠非常有效地使用於脫髮治療。
無
[圖1]示出對毛乳頭細胞中的七葉皂素的Wnt/β-連環蛋白啟動子活性度放大效果的分析結果。
[圖2]示出確認毛乳頭細胞中的由七葉皂素實現的β-連環蛋白的表達量的增加的實驗結果。
Claims (8)
- 一種防脫髮或毛髮生長促進用組合物,其作為有效成分包含七葉皂素,相對於該組合物的總重量,該七葉皂素的含量為1重量%至2重量%。
- 如請求項1所述的防脫髮或毛髮生長促進用組合物,其中,該組合物促進毛乳頭細胞的增殖。
- 如請求項1所述的防脫髮或毛髮生長促進用組合物,其中,該組合物增加毛髮的彈性。
- 如請求項1所述的防脫髮或毛髮生長促進用組合物,其中,該組合物為醫藥組合物。
- 如請求項1所述的防脫髮或毛髮生長促進用組合物,其中,該組合物為醫藥外用組合物。
- 如請求項1所述的防脫髮或毛髮生長促進用組合物,其中,該組合物為化妝料組合物。
- 一種頭髮或頭皮用產品,其包含如請求項1所述的防脫髮或毛髮生長促進用組合物。
- 如請求項7所述的頭髮或頭皮用產品,其中,該頭髮或頭皮用產品選自由生髮劑、頭皮護理劑、頭皮清潔劑、頭皮按摩劑、頭皮保養劑、清洗劑、洗髮水、滋潤劑、護髮素、髮乳、膠、髮膜、霜、精華、粉末、噴霧、油、皂、軟膏、頭髮造型劑、染髮劑及燙劑構成的群組。
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