TW200808368A - Combination - Google Patents
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- TW200808368A TW200808368A TW096106591A TW96106591A TW200808368A TW 200808368 A TW200808368 A TW 200808368A TW 096106591 A TW096106591 A TW 096106591A TW 96106591 A TW96106591 A TW 96106591A TW 200808368 A TW200808368 A TW 200808368A
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- Taiwan
- Prior art keywords
- composition
- skin
- pigmentation
- extract
- treatment
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Description
200808368 九、發明說明: 【發明所屬之技術領域】 本發明係關於包含毛蕊糖苷(verbascoside)及木犀草 素(luteolin)之組合物、含此組合物之植物萃取物及其於化 妝品上或醫樂組合物上供調整色素沈積之用途。
【先前技術】 咖啡酸衍生物毛蕊糖苷為苯丙素糖苷 (Phenylpropanoid glycoside)之鄰二羥基肉桂酸衍生物。苯 丙素糖苷之治療特性上之許多應用已為人所知,例如抗真 菌、抗細菌、抗病毒、止痛劑。 毛,3¾糖普之化學名稱為2-(3\4’-二輕基苯基)乙基 -O-a-L·。比喃鼠李糖基(rhamnopyranosyl)-(l—>3)-(β-Ώ_(4-Ό-咖 啡醯基)-吼喃葡萄糖苷,及其完整的結構係在1963年以洋 丁香苷(acteoside)之名稱說明(Birkofer 等人,Z Naturforsch B,1968,23(8),1051-8)。毛蕊糖苷亦稱為臭梧桐寧 (Kusaginin)且已知其於化妝品上之用途。 毛蕊糖苷於抗老化化妝品組合物上之用途係描述於 W02004/069218中。毛蕊糖苷證實可刺激皮膚細胞之逆境 蛋白(stress pr〇teins)(HSP 70)合成並使皮膚能有效的防禦 5 200808368 環境的攻擊。W02004/069218描述了得自管狀花目,更特 而δ之毛蕊花屬(Verbascum)、(Pladago)、馬鞭草屬 (Verbena)、過江藤屬(1^0叫或斤阳}〇11懦)屬之毛蕊糖脊萃 取物。 5 出現在運動科學1999年第5屆IOC世界大會上之海報 「毛蕊糖苷,一種中藥萃取物,對力竭運動後腦中及骨於 肌中自由基形成之效應」(Κ.Μ· Chan & JX· u),提出了 φ 毛蕊糖苷抗自由基之效應。
毛蕊糖苷亦被論及作為皮膚美白之活性化合物(JP 10 2005-082522)。在wo 01/026670中亦描述來自含毛蕊糖苷 之撖欖植株萃取物之抗老化及皮膚美白活性。 毛蕊糖苷可從市面上購得而萃取毛蕊糖苷之方法亦 有4田述。毛,滅糖皆可付自不同的植物例如玄參科 (Scrophulariaceae)、胡椒科(Piperaceae)、唇形科 15 (Labiatae)、爵床科(Acanthaceae)或列當科 φ (Orobranchaceae),如馬先蒿(Pedicularis)屬(CN1291613)、 樹胡椒(Piper aduncum)(JP2000302797)、米糰花屬 (Leucosceptrum)(JP2191292)、長春藤列當(Qrbranche hedera)(FR2302745) 〇 20 膚色係由荷爾蒙及遺傳決定,但對UV的照射會發生 色素改變。在UV感應後,皮膚顏色主要係藉由黑色素生 成作用來調節。此複雜的生化反應鏈係發生在表皮並對應 由樹狀黑色素細胞產生黑色素。黑色素包括二種多肽:黃 紅色的嗜鉻黑色素(phaeomelanin)及深棕色的真黑色素 6 200808368 (eumelanin)。黑色色生成會受特定的介質影響-如酪胺酸 酶酵素及與酪胺酸酶有關的蛋白(TRP1、TRP2)—其決定 黑色素量及黑色素之種類,因此參與膚色外觀(Petit L,
Pierard GE,2003, 25(4),P.169-181)。 5 此外,膚色之影響因素亦包含從黑色素細胞有效的 將黑色素轉移至鄰近的角質細胞並藉由受體角質細胞將 轉移的黑色素體降解(Boissy RE,及φ〜謂加0/. 2003;12 • Suppl 2:5-12)。在黑色素細胞樹狀形成作用中及/或在含黑 色素之胞器(黑色素體)中扮演一個角色,此轉移亦參與色 10 素沉積之調節。 木犀草素(Luteolin)為一類黄酮分子,其化學名稱為 3’,4’,5,7_四羥基黃酮。
木犀草素 木犀草素係以其在色素沉積上之活性為人所知。 FR2578422申請以生物活性量之木犀草素作典型的 治療:此組合物據賴過度變黑的m療具活性而無毒 !生問,。木犀草素可’例如由洋蓍草(〜仙以millef〇lium) 之乾燥的莖部分萃取。 木犀草素亦被提及具有抗氧化性。DE19962345描述 了 /、有抗氧化特性之化妝品組合物,其包括了至少含 7 200808368 木犀草素之落花生(Arachis hypogaea)種子萃取物。EP 1072265展示木犀草素與其他多酚系化合物組合於抗氧化 活性之用途。 熊果素(Arbutin)因為對胳胺酸酶的抑制而以其在黑 5 色素形成作用上之活性為人所知(Maeda K,Fukuda M, JPharmacol.Exp.Ther”1996, 276, 765-769; Chakraborty and al,Pigment Cell Res, 1998,11(4),206-12)。此對苯二驗 φ (hydroquinone)p-D-吡喃葡萄糖苷因此常在酵素、細胞培 養或活體之實體試驗中用作參照。例如,在目前用於美白 ίο 性質篩選試驗之酵素性蘑菇酪胺酸分析中,熊果素之抗酪 胺酸酶IC50約為 100 pg/ml (Lee KT 等人,Int J Cosmet Sci, 1997, 19(6),291-98; Kang HS 等人,Arch Pharm Res,2004, 27(12)51226-32; Funamyama 等人,Bioscience,Biotechnology and Biochemistry,1995, 59(1),143-44 )。此化合物係如此 15 使用或由植物-例如熊果葉(Uvea ursi folium)衍生(Petit L, 馨 Pierard GE,Int J Cosmet Sci,2003, 25(4),ρ·169-181)—用 於亮化的化妝產品中,因為此對苯二酴衍生物比對苯二酴 更安全,而對苯二酚因為其細胞毒性而禁用於化妝品中。
ΟΗ 熊果素 醉魚草科(Buddlejaceae)係由九個屬(Androya)、醉魚 8 200808368 草屬、(Emorya)、(Gomphostigma)、(Nicodemia)、(Nuxia)、 (Peltanthera)及約150種所組成。 野生腋花醉魚草(Buddleja axillaris Willd)亦稱為 Adenoplusiaaxillaris為一種灌木,高2至5公尺,主要生長 5 於馬達加斯加(Madagascar)及東非之次生林中。對生葉為 單生、有柄固定,長7-12 cm,寬2-4.5 cm ;樹枝上表皮 為綠色及略帶毛狀,下表皮略白及帶有絨毛。花為頂生、 馨聚繳圓錐花序帶有白色花冠,外部有濃密的絨毛而其内為 光滑。果實為棕色、肉質、球形閉果而直徑約為2.5 mm。 10 種子為橢圓體,長約1 mm。在馬達加斯加當地此種植物 稱為「Sevafotsy」或「Mandresy」,傳統上係作為保健之 用,例如用水煎煮係作為治療頭痛之飲料,而包含葉子和 樹幹煎煮液以及一些煮過的植物之混合物係用作對抗風 濕病及關節炎之糊劑。 15 日本專利已有描述革葉醉魚草(Buddlej a coriacea)萃 _ 取物單獨或與另一種植物萃取物組合用於美白組合物中 (JP5225062, JP8012565)。一特異的類黃酮分子稱為類醉魚 草素(Buddlenoid)係揭示作為活性化合物(JP5255376)。 密蒙花(Buddleja officinalis)已有研究。已由密蒙花 20 之花朵中分離出四種類黃S同、一種苯乙基糖苷及一種苯丙 素糖苷。在這些分子中,木犀草素及洋丁香苷(=毛蕊糖苷) 已顯示具有抗氧化特性(Piao MS, Kim MR,Lee DDG,
Park Y,Hahm KS,Moon YH,Woo ER,Arch Pharm Res, 2003 Jun,26(6),453-7) 〇 9 200808368 FR2831444指出包含大葉醉魚草(Buddleja davidii)及 創傷絨毛花(Anthyllis vulneraria)水溶液萃取物之化妝或 皮膚用組合物。此組合物聲稱對太陽曝曬後之皮膚修復具 有濕潤、緩和、抗刺激及癒合傷口之特性。此水溶性的醉 5 魚草萃取組合物經描述並含有環稀醚萜(iridoid)、類黄 酮、咖啡酸酯及三萜類(triterpenoid)。 毛蕊糖苷已從其他種之醉魚草科,例如從(Buddleja • yunanesis)(Liao YH 等人,J Nat Prod,1999, 62(9),1241-5) 或從甘肅醉魚草(Buddleja purdomii)(Gao Y 等人,Zhong 10 Yao Cai,2004, 27(5),339-41)中分離及辨識出。由心葉醉 魚草(Buddleja cordata)(Avila Acevedo JC 等人, Fitoterapia,1999,66(1),75-78)及由球葉醉魚草(Pardo F 等人,J of Ethnopharmacology,1993,39(3),221-2)中所分 離出之毛蕊糖苷已顯示具有抗菌活性。再者,毛蕊糖苷可 15 由召澤醉魚草(Buddleja scordioides)中分離出(Avila φ Acevedo JC 等人,Fitoterapia,2005, 76(3-4),301-309)。 【發明内容】 本發明係關於包含毛蕊糖苷及木犀草素之組合物, 20 及/或含植物萃取物之組合物供調整色素沈積之用。使用 根據本發明組合物及含萃取物之組合物為適當及安全之 調整皮膚色素沉積之方法。 根據本發明含毛蕊糖苷之植物萃取物係包括(但不 限於)管花科包括例如,(Verbascum)、(Pladago)、 200808368 5
10 15
20 (Verbena)、(Lippia)或(Fraxymus)屬;醉魚草科包含例如, (Androya)、醉魚草屬、(Emorya)、(Gomphostigma)、 (Nicodemia)、(Nuxia)或(Peltanthera)屬;或脣形科包含例 如,巴落草(Ballota)、(Faradaya)屬。較佳的係給予醉魚草 屬而更佳地係為腋花醉魚草之植物萃取物。 萃取可在植物的所有部位進行。較佳的係由腋花醉 魚草之葉片萃取。 萃取可以標準的萃取方法來進行。較佳地萃取係以 可用於萃取之極性溶劑來進行。先以極性溶劑萃取葉片, 視要可萃取數次。然後將所得到的溶液混合並以非極性 溶劑例如庚烷萃取以移除蠟、精油、色素及大部分的非極 性分子。進行相分離後,將留在極性層之溶劑移除以得到 含毛蕊糖苷之乾燥萃取物。視需要,萃取物可藉由加入水 並進行冷凍乾燥來乾燥。 本發明萃取物一般為乾燥的萃取物。然而,此萃取 物亦可以浴液來使用,亦即省略所述之萃取方法中最終的 摩文呆步驟。 入用於萃取之極性溶劑較佳地為醇類或水與醇類之混 二物二其中醇類較佳的為乙醇。水與醇間之體積比可由 50.50兩至9〇:1〇,較佳地70:3〇。 較佳_給予的含毛蕊料之乾驗物萃取物之量 j於ω/° ’較佳地係大於15%,最佳地為16%至25%, 舌旦犀草素之里⑥至5%,更佳地高至,最佳地高至斤〇 1比之總植物萃取物。植物萃取物含有木犀草素之量至 11 200808368 少為0·01 %重量比之總植物萃取物。最佳地,植物萃取物 為腋花醉魚草之萃取物。 令人驚訝地,相較於僅投予毛蕊糖苷,只需要少量 的木犀草素即可得到較佳的皮膚美白效果。 組合物可為協同性,其中藥物之聯合作用係使該組 a的效應大於其個別效應之代數總和。因此,可投予減量 之藥物,例如降低毒性或其他有害的或不欲的效應,及/ 或使用與單獨投予該藥物時相同之量,但達到較大的功 效。減量之藥物可以較低量用於標準方法中,其中例如投 予單一藥劑。 本發明之組合物可在任何時間及以任何有效的形式 來給藥。例如,例如化合物可同時給藥,例如以單一組合 物或劍1單位(例如片劑或包含二者組合物之液體),或其 可以分開的組合物但於相同的時間給藥(例如其中一種藥 係以靜脈給藥而另一種則以口服或肌肉内給藥)。藥物亦 可於不同的時間連續給藥。藥劑可依習用調配以在_段延 長的時間内(例如12小時、24小時)達到所欲的釋放速率。 /、了藉由使用具適合的代謝半衰期之藥劍及/或其衍生物 來達成,或藉由使用控制釋放調配物來達成。 包含毛蕊糖苷及木犀草素之組合物可從含有彼等物 貝之萃取物以標準的分離方法來分離及/或純化。標準的 分離方法包括但不限於層析法。 、 本發明之組合物或包含該物質之萃取物可為任何形 式以任何有效的路徑來給藥,包括口服、非經腸、腸内二 12 200808368 评脈内、腹腔内、局部、皮膚滲透(例如使用任何標準的 貼布)、眼部、鼻内、局部、非口服,例如氣霧、吸入、 =肉内頰内、舌下、直腸、陰道、動脈内及鞘内 5
10 15 專,。其可單獨或與任何成份(具活性或不具活性)組合來 給樂。較佳的係為局部給藥。 本發明之組合物$包含該組合物之萃取物可以已知 ^方法轉變成常用之調配物,例如化妝品、皮膚及/或醫 樂組合物。此等可為液體或固體調配物,例如(不限於)一 般及腸衣錠劑、膠囊、片劑、散劑、顆粒、酏劑、酊劑、 溶液、軟膏、乳劑、凝膠、油膏劑、漿液、泡沫、洗髮劑、 條劑或乳液。 較佳的係給予水溶液、白色或有顏色的乳霜、軟膏、 乳劑、凝膠、油貧劑、漿液、泡沫、洗髮劑、條劑、乳霜、 糊劑或乳液形式之皮膚或化妝品組合物。 本發明之組合物或包含該組合物之萃取物可進一步 與任何其他適合的添加劑或醫藥上可接受載劑組合,較佳 地為皮膚及/或化妝品上可接受載劑。此等添加劑包括任 何已提及之物質,以及任何習用之物質,例如Remington·· The Science and Practice of Pharmacy (Gennaro及 Gennaro, eds5 20th edition, Lippincott Williams & Wilkins, 2000); Theory and Practice of Industrial Pharmacy (Lachman 等人, eds·, 3rd edition,Lippincott Williams & Wilkins,1986); Encyclopedia of Pharmaceutical Technology (Swarbrick及 Boylan,eds·,2nd edition,Marcel Dekker,2002)中所述之物 13 20 200808368 質。這些本文中所指之「醫藥上可接受載劑」係指其可與 活性藥劑組合及能以治療為目的安全地投予一對象者。 本發明之組合物或包含該組合物之萃取物之劑量可 參照所欲治療之效用及/或疾病種類及/或疾病狀況來選 5 擇,以提供所欲的治療活性。這些劑量例行上係由特定病 患之各種可用於選擇適當劑量之參數來決定(例如疾病的 種類、病患的年齡、疾病狀況、病患的健康、體重等)或 φ 該量可為相對上之標準。 所投予的活性成份之量可根據此等考量,如所用的 10 特定化合物和劑量單位、投藥模式和時間、治療期間、年 齡、性別和所治療病患之一般狀況、所治療症狀之性質和 程度、藥物的代謝和排出速率、潛在的藥物組合和藥物-藥物之相互作用等廣泛地變化。 較佳的所給予之組合物含有毛蕊糖苷之量至少為總 15 組合物之0.0001%,較佳為總組合物之至少0.001%重量 • 比。亦較佳的所給予之組合物含有毛蕊糖苷之量高至 10%,較佳地高至5%,更佳地高至1%重量比。亦較佳的 所給予之組合物含有木犀草素之量為高至總組合物之 1%,,更佳地高至0.1%,最佳地高至0.05%重量比。亦較 2〇 佳的所給予之組合物含有木犀草素之量為總組合物之至 少0.00001%高至1%重量比。 本發明之組合物可包含本發明之乾燥的植物萃取物 之量為總組合物之0.01%高至10 %,較佳地0.1%高至1%重 量比。 14 .200808368 次,更佳地高—或多次,較佳地高至三 然而,在某此情也係以局部給藥。 表現、製備物之種類==照體重、對活性成份之個別 里分開’可㈣有利。例如,在 低量可能就足夠,而在1仙卜主、〇 ' rm^、別述之取
10 15 就投予相t大量之情況下下則必須超過所述上限。 數個劑量。^下奸係將這些量分成—天内之 本4明之組合物或包含該組合物之萃取物亦可與至 少-種其他活性物質或包含常用於皮膚用途之物質之萃 取物,合。其他的活性物質包括(但不限於)供皮膚美白 用、宂化皮膚、預防斑點或治療斑點之物質,例如對苯二 酚、維生素Α酸(tretinoin)、局部用類固醇、杜鵑花酸(azdic acid)、麴酸(kojic acid)、熊果素、木犀草素及甘草(Hc〇rice) 萃取物。 適合用於色素沈積調節之物質如防曬劑或過溏 劑、角質溶解劑例如α-羥酸,亦可與本發明之組合物或包 含該組合物之萃取物組合。 本發明之組合物或包含該組合物之萃取物可用於皮 膚科領域,其包括化妝品及醫藥上用於色素沈積調節。 本發明之組合物或包含該組合物之萃取物可用於化 妝品上供皮膚美白、亮化皮膚、預防及降低皮膚之色素斑 點(老化或光引起之斑點)、抗皮膚之色素沉積、均化皮膚 色調及/或白皙皮膚。 15 200808368 本發明之組合物或包含該組合物之萃取物亦可用於 預防治療、預防或調節色素沉積病症,其包括但不限於傷 口癒合後發炎後色素沉積(痤瘡、溼療、觸性皮膚炎等)、 光黑色素生成、内分泌異常及懷孕(痞)、艾迪生氏症、黑 5 棘皮病、雀斑、黑皮病、抗生素之副作用、抗瘧疾劑治療、 在皮膚治療期間預防癌細胞自我保護、漸進性色素沉積紫 斑、預防老人班及因使用化妝品(例如香水等)所引起之色 _ 素沉積。 1〇 本备明之組合物或包含該組合物之萃取物顯示影響 酪胺,酶、黑色素形成、uv引起之色素沉積、與色素沈 積凋即有關之黑色素細胞樹突形成及/或黑色素體轉移。 【實施方式】 取物之.備 響物萃壓碎的腋花醉魚草㈣以7G : 3Ό乙醇及水之混合 可ί清°屮將此溶液在萃取步驟期間攪拌及加熱。固體物質 日士、及可重複萃取數次。萃取時間為30分鐘至1小 叶。溫度為6〇〇Γ 2〇 庚烷萃取。^以下。然後將組合的乙純萃取液混合並以 移除溶劑。進二相分離後將剩餘的極性層於真空下蒸餾, 怒馇註二&現需要加入水進行冷凍乾燥以得到最終的含毛 _搪杳之摩乞燥萃取物。 將最终Μ * ρ ^ 定性。最終的—乾紐卒取物以薄層層析及HPLC標準方法 ;的萃取物顯示含有以乾燥萃取物的總重量計 16 200808368 19%之毛蕊糖苷及0.1%之木厚草素。 實例2·酿胺酸酶抑制任掉 5
10 15
20 根據實例1所製造之腋花醉魚草萃取物之抑制活性 係於活體外進行評估。此方法係以測量因多巴色素 (dopach贿e)之作用在475腿所增加的吸收光度所測定磨 菇酪胺酸酶之多巴_氧化酶活性為基準。 ^汁异抑制濃度IC50(試驗產品降低5〇%對照的酪胺酸 酶之多巴氧化酶活性時之濃度)。 數據係以每分鐘吸收度的變化來表示(AA/At)。 、將醉魚草萃取物直接溶於分析緩衝液中(磷酸缓衝 液)進行5種濃度之試驗:〇.03mg/ml、〇·ΐ〇 mg/ml、0·30 1 mg/ml及3 mg/ml。各實驗條件係以二重複進行。 貝驗翏數為:40 U/ml之酵素濃度及4分鐘内於475 nm所測 量之吸收度。 產品之酪胺酸酶抑制作用係如表丨所示。考慮抑制百 为比於试驗產品濃度之線性關係(以對數(一)表示),由下 列回歸曲線計算抑制劑量(1C50):抑制% 2.57 1〇g(濃度) + 67.43 ddl5 r-0.976) 试驗產品之IC5〇為290 pg/ml。 17 200808368 濃度 (mg/ml) AA/At 酪胺酸酶 (U/ml) 抑制% 0 0.151 58.6 - 0.03 0.126 48.6 16.9% 0.1 0.092 35.5 39.4% 0.3 0.076 29.5 49.6% 1.0 0.063 24.2 58.7% 3.0 0.017 1-- ---------- 6.6 88.8% 實例3....腋花醉魚草萃电A對皮膚色音率统之效用 5 為了評估色素沉積之活性,在試驗化合物之存在 下,以測量暴露(UVB-刺激)或未暴露在放射線下之培養的 人類黑色素細胞之胞内黑色素含量為基準,進行一活體外 •實驗。進行腋花醉魚草萃取物(根據實例1}、毛蕊糖苷(純 度>90%,Extrasynthese公司)及木犀草素(自腋花醉魚草中 ίο 分離出)之試驗。 將得自包皮之正常的人類表皮黑色素細胞於30〇c 下,於添加抗生素之MGM「無血清」培養基(黑色素細胞 生長培養基,PromoCell®)中培養。培養係維持在37〇c及 濕化的5% C02氣壓。 15 將次代培養之人類黑色素細胞轉置於MGM培養基 18 200808368 中’並在細胞剛好聚滿前使用。計算細胞數並於無牛血清 之培養^中稀釋至所欲的濃度。將培養的黑色素細胞以每 孔60X103細胞之密度置於24-孔盤中。於二個測定盤上接 種細胞丄—個供測量黑色素濃度(”黑色素,,盤,,)而另一各 5 則供測量細胞密度Γ中性紅(Neutral Red)”盤)。 ° 以UVB處理: • 、,盤72小時後,移除培養基並以含試驗產品無毒之 各種濃度的新鮮培養基取代之。於37〇c、95%空氣- 10 氣壓下將細胞培養8天。在此期間以第1天(1)1)、第2天 (D2)、第3天(D3)、第4天(D4)、第7天(D7)及第8天(D8)將 細胞暴露在UVB照射下。UVB照射係以連續發射帶有312 nm^射冋峰之280至320 nm光譜之平行TL20W/12燈管組 來進行。每次照射係提供4〇mj/cm2之UVB量。在無而暴 15 露及無處理下將偽劑一對照組細胞進行相同的程序。 φ 在照射前以預熱過的磷酸緩衝溶液(PBS)清洗細胞 單層並在PBS的存在下(無試驗產品)暴露mUVB下。在照 射後立即以新鮮的試驗培養基置換。 说 试驗階段(D9)終了之處理: 處理後將培養基移除。以pH 6.8之PBS清洗細胞並加 入NaOH-DMSO溶液萃取黑色素。將黑色素萃取液於8〇£>c 加熱2小時。冷卻後,將各等份加到96孔之微量測定盤中。 以微盤判讀機(Dynatech MR 5000)於450 nm記錄光學密度 19 200808368 (OD450nm) 合成的黑色素標準(Sigma)係於相同%% 此標準曲線能將。公㈣疆轉變成黑色素單 1件下培養。 、· , Α/π 、 ^§Kmelanin unit equivalent)/孑L 0 5 黑色素的含量係以由對照組及經處理的黑色素笑取 物於405 rnn之吸收度來測量。結果係以每孔之叫專色^來 表示,由標準曲線來測定:DO(〇D45〇nm)二f([黑色素]
10 結果: 在前述處理及UVB暴露培養後,測量人類正常黑色 素細胞(胞株Μ99-〗!!6)之胞内黑色素含量。 進行3種濃度之各試驗化合物之研究:lHg/ml、$ pg/ml及 10 pg/ml。 15 細胞數評估(以中性紅吸收法)顯示3個試驗劑量皆無 產品毒性。 … 以各對應培養之細胞密度(每孔之細胞數)校正黑色 素含量(每孔pg之黑色素),以表示出細胞之「黑色素形成 量」(以pg黑色素/細胞表示)。 根據下式计异試驗產品對色素沉積活性: (黑色素含量/細胞) 經處理一 (黑色素含量/細胞) 對照組 FA= — -----— X 100 (黑色素含量/細胞) 對照組 20 200808368 表2: 試驗 黑色素含量 (Pg黑色素/子L) PA (%) 對照組 無UV 對照組- 50.34 +/- 0.93 - 有UV 對照組+ 74.10+/-1.59 • 腋花醉魚 草萃取物 有UV 1 jig/ml 61.14+/-0.87 ΛΊ%(ρ<0·01) 5 pg/ml 58.85+1-0.47 -2\%(ρ<0Μ1) 10 jug/ml 59.61 +/- 0.00 -20%(ρ<0·01) 木犀草素 1 jug/ml 57.56 +Λ 0.0 ^22%(ρ<0Μ1) 5 jig/ml 55.57+/-0.0 -25%(ρ<0Μ1) 10 pg/ml 54.84+/- 1.56 -26%(p<0.01) 毛蕊糖苷 10 pg/ml 59.16+Λ 1.56 ^20%(p<0.01) 50 pg/rnl 58.32 +A 4.44 -2ί%(ρ<0.01) 100 pg/ml 57.80+/- 1·63 ^22%(p<0.01) 當經每日相當UVB曝曬之黑色素細胞以三種試驗產 5 品(表2)培養時,胞内黑色素含量明顯的降低。 試驗產品對經UV刺激之黑色素細胞之黑色素形成 具有明顯的抑制效用:相較於經照射之控制組培養及同等 物3種濃度之各產品(p^O.Ol,學生試驗)其黑色素含量明 顯下降。 21 200808368 實例4.調配物U勾膚菝液) INCI名稱 量(g) 甘油 3.00 丙二醇 2.00 實例1之腋花醉魚草葉萃取物 1.00 辛水揚酸(Octocrylene) 0.20 苯氧乙醇+對羥基苯曱酸曱酯+對羥 基苯甲酸乙酯+對羥基苯甲酸丙酯+ 對羥基苯甲酸異丁酯 0.80 卡波姆(Carbomer) 0.50 EDTA四鈉 0.10 氫氧化納 qs pH 5.5-6 水 qs l〇〇g 實例5.調配物2(抗色素沉積乳霜) INCI名稱 量(g) 鯨蠟醇(及)硬脂酸甘油酯(及)PEG-75 硬脂酸(及)Ceteth-20 (及)Sterateh-20 6.00 PPG-115硬脂醯醚 4.00 甘油 3.00 22 200808368
丙二醇二壬酸酯 2.50 __-- 異十六烧 2.40 __— 辛喊 1.20 —--- 十六酸異丙酯 L20 __一 丙二醇 L00 —一 實例1之腋花醉魚草葉萃取物 0.50 __.— —甲基梦氧烧(Dimethicone) 0.50 __-—一 苯氧乙醇(及)碘代丙炔基胺甲酸丁酯 0.50 —-—一 亞硫酸氫鈉 0.15 _-一 EDTA 二鈉 ----- 0.05 _—* 抗壞血酸 '----- 0.05 氫氧化納 ~~·~———____ qs pH 5-6 水 qs IOOg S __ __·-—J 之調配目標係顯示含0.5%腋花醉魚草萃取物(實例5) 膚色素沉積品,對亞洲自願者於因UVA照射所㈣ 劑,但含、果素二二:物包含與實例5相同之賦形 23 5 200808368 方案: 表3 : 處理 包括之自願 者人數 —---- 完成此研究之 自願者人數 皮膚光照型態分布 數據分析所包括 之自廟表,土, 具0.5%腋花 醉魚草萃取 物之乳霜 8 7 5皮膚受光型態III 2皮膚受光型態IV -數 6 具1%煦果 素之乳霜 8 ——_ 8 -~---- 6皮膚受光型型態III 2皮膚受光型態IV 8
5
^對各產品而言,此研究為一開放、個體内之研夂 個者為其本身之對照。此研究細平行群組來進^ 個群組一種產品)。 ( 试驗開始前14天每天塗抹乳液二次(早、晚),並 ”究後1G天(D〇至⑽)。乳液係以正常的使用狀況塗ς 於背部治療區域,亦即使用賴完全定位該區,由第三者 以按摩的S式塗抹乳液直到產品滲透。處理配置及產品塗 =侧(右/左)為隨機的。膚色之測量係於D〇、D2、D4及 时進仃。於DO、D2及D4時進行照射(1 MPD二最低色素沉 積之劑量)。 、 ^ UVA照射係以裝有消除紅外線(IR)及可見光濾光器之 氙氣短弧光燈(Arquatiel Idem 2000,光譜·· 320-400 nm)來 進行。 皮膚色素沉積進展之評估(有或無塗抹產品之UV所引 15 .200808368 起的褐邊強度)係使用CR321 Minolta⑧Chromameter®色度 計藉由測量色度來進行。Chmmameter®可將顏色轉變成由 二位参數所組成之數位瑪:L* :為明亮度(由深至淡),a* : 為綠至紅之色譜,b* :為藍至黃之色譜。a*&b*為色度參 數而L*為亮度參數。此儀器常用於化妝品及醫藥上皮膚顏 色之測量。 最具特徵性之色素沉積參數為黃色(b*)及亮度(L*)。亮 度L·*增加係反應色四素沉積強度降低。b*增加其特性為皮 膚之黃色成份增加以及色素沉積強度降低。 二種參數係沉積程度ITA。(個體拓樸角(Individual Topologic Angle)之計算,ITA。係根據下式整合明度(L*)及 黑色素形成參數(b*)定義受試者皮膚色素沉積程度: ITA°=(Arc TAN((L*-50)/b*))xl80^ ITA增加其特性為色素沉積強度降低。 結果: 色度參數L*、b*及ITA在處理及未處理區之變化(△)係 根據下式來計算: A=(TZt 厂 TZt0) - (NTZt「NTZt0) 其中: TZ :處理區域所得到之值 NTZ :非處理區所得到之值 25 .200808368 t〇 : 塗抹產品前 ύ: 塗抹產品後之每次測量時間 由各自願者中得到之任意單為(A.U.)或度(。)之變 化,以及敘述性統計係如下表所示。 5 表4 · UV照射後以及重複塗抹含〇 5%腋花醉魚草萃取 物產品(τι)之膚色變化。與非處理區(ΝΤ)之比較
10 動力學 (n=7) ~——_ 研究參數 原始變化 Tl/NT (moy 士 SEM) 具有色素沉積 抑制之自願者 人數 顯著性 (ANOVA) ㈣) D2/D0 L* +2.01±0.64 6/7 有(pHXOll) b* -〇.81±〇.3l —-------- 6/7 限制(P=〇.〇85) ITA° +7±2 6/7 —--- 有(@=0.007) D4/D0 1—-—-_ L* +138±〇55^ 6/7 有(ρ=0·009) b* -0.51 士 〇·41 —--—_ 5/7 限制(ρ=〇、〇93) ITA° ————-- +5±2 -—--- 5/7 有(ρΗ).006) 以人讀的志願者㈣,由UVA照㈣生褐變後 :草萃取物之乳液處理之區域明顯地 相二理區具較少的色素沉積(表4)。觀察到之1TA。增加 目^非處理區,+7。及+2。分別於d2ad4(p:oo〇7 ^ 了 IMG,處理及麵理區並無顯著之差異。然而 ΐί甘而言似乎褐變係在照射後第六天開始 ,、可_此項結果。含Q 5%職醉魚草萃取物 26 15 200808368 產品明顯地抑制了由UVA所引起的皮膚色素沉積直到 D4 〇 表5 : UV照射後以及重複塗抹含1%熊果素產品(T2) 之膚色變化。與非處理區(NT)之比較 動力學 (n=7) ---- 研究參數 — 原始變化 T1/NT (moy 土 SEM) 具有色素沉 積抑制之自 願者人數 顯著性 (ANOVA) ㈣) D2/D0 +0.32 土 0.48 4/8 無(ρ=0·389)一_ -0·3 8土0·34 5/8 有(Ρ 二 0.019) ------- ITA° —-—— +2 士 2 4/8 無(ρ=0·220) D4/D0 L* +0·48 土 0·34 5/8 無(ρ=0·534) b* -0.82土0.3 7 7/8 無(Ρ=0·278) —-~~~-_____ ITA° +4 士 1 7/8 限制(Ρ=〇·〇60) _ 對大多數的志願者而言,以二次UVA照射後,以含 1G/〇熊果素之產品處理之區域明顯地比非處理區具較少的 色素 >儿知(與非處理區相比,8位自願者中有7位在D4 其ITA增加+4。,ρ=〇·〇60,表4)。在一次和三次照射後 (D2及D10),抑制效用較低(8位自願者中有4位其ΙΤΑ。 增加+2。,分別為ρ=〇·220及0.363,表4)。 15 對大多數的志願者而言,含1%熊果素之產品傾向抑 制因UVA所引起之皮膚色素沉積(變化在D4其顯著性有 限)。 27 200808368 實例7.於黑色素細胞樹突形成之活體外活性 此研究係以評估含0.5%腋花醉魚草萃取物(實例5, P) 與賦形劑(E)之乳液之抗色素沉積活為目標。 此次活體外實驗係使用由腹部整形手術得來之切片 5 (27歲,皮膚受光型態ΙΙ/ΠΙ)。將其培養於特定成活外植 體培養基BEM(BIO-EC之外植體培養基)並根據其特定處 理分配:
φ 對照組 C
未經光照之控制組外植體 C-UV
1〇 經光照之控制組外植體 C+UV 未照光之外植體,以賦形劑+0.5%
腋花醉魚草萃取物處理 P-UV
未照光之外植體,以賦形劑處理 E-UV 經光照之外植體,以賦形劑+
15 0· 5%腋花醉魚草萃取物處理 P+UV
. 經光照之外植體,以賦形劑處理 E+UV 將2 mg的產品(P,E)局部塗於外植體並以小的壓舌片 抹開。這些係於第 0 天(DO)、Dl、D2、D3、D4、D5、D6、 20 D7及D8時塗抹於各處理的外植體。. 外植體C-UV、P-UV及E_UV未經光照。外植體 C+UV、P+UV 及 E+UV 每天接受光照(UVA : 2,25 J/cm2, UVB 0.135 J/cm2)。光照係於局部塗抹賦形劑或賦形劑 +0.5%萃取物前2小時進行。在其放回BEM(BIO-EC之外 28 200808368
植體培養基)後,於光照期間將外植體之培養基換成HBSS 缓衝液。 於D3、D6及D9時將外植體取出進行組織學研究。 母-人皆將外植體切成3個部分。一部分定於甲酸溶液中, 5 其他的部分則凍存於-80°C。 根據馬森氏法(Masson’s method)將經甲醇固定之外植 體脫水、以石臘浸塗及染色後,進行一般型態學研究。 φ 根據Laidlaw及Blackberg法處理外植體以進行多巴 (DOPA)-氧化酶反應。此項技術能就地評估產品對酿胺酶 10 之活性。 D0 :黑色素細胞具中度d〇PA-陽性。 D3 : P+UV外植體黑色素細胞具輕度DOPA-陽性,陽性明 顯地比經未處理之控制組低。其樹突輕微下降。 E+UV外植體黑色素細胞為明顯的DOPA-陽性及具 15 輕微的樹突狀。 φ D6 :所有的外植體(C+UV、P+UV、E+UV)明顯地為 DOPA-陽性。然而E+UV之黑色素細胞為明顯地樹 突狀,P+UV細胞之樹突輕微地降低。 D9 : P1+UV黑色素細胞為輕微的DOPA-陽性,明顯地低 2〇 於C+UV。樹突明顯地減少。P2+UV之黑色素細胞 為顯明的DOPA-陽性及非常明顯的樹突狀。 產品P1使DOPA-陽性降低。此效用於9天後更容易 觀察到。根據此研究,黑色樹細胞樹突之降低非常明顯且 從第6天及UV照射後可觀察到。 29 200808368 產品E無法對D Ο PA陽性或黑色素細胞樹突造成任何 改變。 在這些操作條件下,相對於在未經處理的外植體中所 觀察到的,此等結果顯示產品Pl(賦形劑+0.5%腋花醉魚草 5 萃取物)具有明顯的抗色素沉積/亮化活性。
30
Claims (1)
- 200808368 十、申請專利範圍: 1 · 一種包含毛蕊糖苷(verbascoside)及木犀草素(iute〇lin) 之組合物。 2· 如申請專利範圍第1項之組合物,係與至少一種其他 活性物質或含有通常用於皮膚用途之物質之萃取物組 合010 15 3· 如申請專利範圍第2項之組合物,其中該其他物質係 為美白皮膚、亮化皮膚、預防斑點或處理斑點之物質。 4· 如申請專利範圍第2項之組合物,其中該其他化合物 係由下列組成之群中選出:對苯二酚、維生素A酸 (tretinoin)、局部用類固醇、杜鵜花酸(azeiic acid)、麴 酸(kojic acid)、熊果素及甘草(licorice)萃取物。 5. 一種植物萃取物,其包含如申請專利範圍第1至5項 任一項之組合物。 6· 如申請專利範圍第5項之植物萃取物,係含有以植物 萃取物之總重量計高於10%之毛蕊糖苷量及至高5% 之木犀草素量。 7· 如申請專利範圍第5至6項中任一項之植物萃取物, 其中該植物萃取物為腋花醉魚草之萃取物。 8· 一種包含如申請專利範圍第1至7項中任一項之組合 物或植物萃取物之組成物,係用於皮膚美白。 9. 如申請專利範圍第8項之組成物,其中該組成物係為 一供局部給藥之皮膚用組合物。 10. 如申請專利範圍第8至9項中任一項之組成物,其中 31 20 200808368 該組成物為液態溶液、軟膏或乳霜。 11.如申請專利範圍第8至10項中任一項之組成物,係含 有組成物之0.0001%至10%重量比之毛蕊糖苷及 0.00001%至1%重量比之木犀草素。 5 12. 一種如申請專利範圍第1至11項中任一項之組合物或 萃取物或組成物於製造供調節色素沉積之組成物之用 途。 φ 13.如申請專利範圍第12項之用途,係用於影響酪胺酸 酶、黑色素形成、UV引起之色素沉積、與色素沈積調 10 節有關之黑色素細胞樹突形成及/或黑色素體轉移。 14.如申請專利範圍第12項之用途,係用於美白皮膚、亮 化皮膚、預防及降低皮膚之色素斑點、預防或降低與 老化有關或光引起之皮膚斑點、抗皮膚之色素沉積、 均化皮膚色調及/或白皙皮膚。 15 15.如申請專利範圍第12項之用途,係用於治療、預防或 I 調節色素沉積病症。 _ 16.如申請專利範圍第15項之用途,係用於治療、預防或 調節選自傷口癒合後發炎後色素沉積、光黑色素生 成、内分泌異常及懷孕(痣)、艾迪生氏症、黑棘皮病、 2〇 雀斑、黑皮病、抗生素之副作用、抗癔疾劑治療、在 皮膚治療期間預防癌細胞自我保護、漸進性色素沉積 紫斑、預防老人班及使用化妝品引起之色素沉積之色 素沉積病症。 32 200808368 七、指定代表圖: (一) 本案指定代表圖為:第(無)圖。 (二) 本代表圖之元件符號簡單說明: 無 t 八、本案若有化學式時,請揭示最能顯示發明特徵的化學式: 15 無20 25
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CN (1) | CN101394900A (zh) |
AR (1) | AR059545A1 (zh) |
CA (1) | CA2644045A1 (zh) |
PE (1) | PE20080135A1 (zh) |
RU (1) | RU2008138390A (zh) |
TW (1) | TW200808368A (zh) |
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-
2007
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- 2007-02-21 KR KR1020087023541A patent/KR20080097487A/ko not_active Application Discontinuation
- 2007-02-21 WO PCT/EP2007/001459 patent/WO2007098873A1/en active Application Filing
- 2007-02-21 RU RU2008138390/15A patent/RU2008138390A/ru not_active Application Discontinuation
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- 2007-02-21 JP JP2008556687A patent/JP2009529499A/ja not_active Withdrawn
- 2007-02-21 EP EP07703526A patent/EP1996295A1/en not_active Withdrawn
- 2007-02-21 CN CNA2007800070479A patent/CN101394900A/zh active Pending
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CN101394900A (zh) | 2009-03-25 |
EP1996295A1 (en) | 2008-12-03 |
PE20080135A1 (es) | 2008-04-06 |
AR059545A1 (es) | 2008-04-09 |
WO2007098873A1 (en) | 2007-09-07 |
KR20080097487A (ko) | 2008-11-05 |
US20090028969A1 (en) | 2009-01-29 |
JP2009529499A (ja) | 2009-08-20 |
CA2644045A1 (en) | 2007-09-07 |
UY30173A1 (es) | 2007-09-28 |
RU2008138390A (ru) | 2010-04-10 |
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