TW201141515A - Antiandrogen agent and sebum secretion inhibitor - Google Patents
Antiandrogen agent and sebum secretion inhibitor Download PDFInfo
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- TW201141515A TW201141515A TW099145527A TW99145527A TW201141515A TW 201141515 A TW201141515 A TW 201141515A TW 099145527 A TW099145527 A TW 099145527A TW 99145527 A TW99145527 A TW 99145527A TW 201141515 A TW201141515 A TW 201141515A
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- lactoferrin
- antiandrogen
- agent
- active ingredient
- test
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- VSIVTUIKYVGDCX-UHFFFAOYSA-M sodium;4-[2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)tetrazol-2-ium-5-yl]benzene-1,3-disulfonate Chemical compound [Na+].COC1=CC([N+]([O-])=O)=CC=C1[N+]1=NC(C=2C(=CC(=CC=2)S([O-])(=O)=O)S([O-])(=O)=O)=NN1C1=CC=C([N+]([O-])=O)C=C1 VSIVTUIKYVGDCX-UHFFFAOYSA-M 0.000 description 1
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- 241000894007 species Species 0.000 description 1
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- 239000012086 standard solution Substances 0.000 description 1
- 239000003270 steroid hormone Substances 0.000 description 1
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- 239000006228 supernatant Substances 0.000 description 1
- 239000011975 tartaric acid Substances 0.000 description 1
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- 210000001635 urinary tract Anatomy 0.000 description 1
- 235000019154 vitamin C Nutrition 0.000 description 1
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Landscapes
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- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Description
201141515 六、發明說明: 【發明所屬之技術領域】 本發明係有關抗雄激素劑、及含該抗雄激素劑爲有效 成分之皮脂分泌抑制劑、養髮劑。 【先前技術】 許多類固醇激素自產生的臟器以被分泌的分子型與受 器結合而表現其作用,而總稱爲雄激素的男性激素,於例 如腦下垂體、腎上腺、睾九等生成之睾固酮,進入目標臓 器的細胞內之後,藉由睪固酮5-甲型還原酵素(5-alpha reductase)被還原爲5-甲型二氫睪固酮(DHT)後與受器 結合,表現雄激素之作用。 雄激素係重要的荷爾蒙,但過度作用時則會誘發男性 禿、多毛症、脂漏症、痤瘡、***肥大、***腫瘤、 男童性早熟等等各種令人困擾的症狀。因此認爲藉由抑制 過多的雄激素的作用,可改善該等令人困擾的症狀,可行 的方法係藉由阻礙將睪固酮還原爲活性型DHT之睪固酮5-甲型還原酵素之作用,抑制活性DHT生成之方法,及藉由 阻礙由睪固酮生成之DHT與受器結合,使其無法表現作用 之方法。 因此,此次藉由睪固酮5 -甲型還原酵素阻礙試驗、雄 激素受器結合阻礙試驗、及進而使用培養倉鼠皮脂腺細胞 之皮脂腺功能抑制評價法,針對數種天然素材進行篩選, 結果發現存在於哺乳動物之乳汁、淚液、唾液、膽汁等分 -5- 201141515 泌物中之乳鐵蛋白,具有高抗雄激素作用,及抑制皮脂分 泌的可能性。 至今已進行許多有關乳鐵蛋白之硏究。由該等結果可 得知乳鐵蛋白具有藉由螯合作用自葡萄球菌、大腸桿菌奪 取鐵質,抑制細菌及病毒的繁殖;使乳酸菌及比菲德氏菌 等益菌增生;提高鐵質吸收預防貧血;活化自然殺手細胞 等免疫細胞,提高免疫力;減輕生理痛,預防骨質疏鬆, 安定情緒;促進τ細胞活化使IgE抗體降低,減少自肥大細 胞產生的組織胺,緩和花粉症等過敏症狀;改善口腔潰瘍 、口臭及香港腳等:使血清中對抗C型肝炎的「介白素-18 」上升;保護黏膜,緩和乾眼症及乾嘴症等症狀;抑制致 癌性物質生成,預防肝癌及大腸癌等癌症;抑制弓漿蟲等 原蟲繁殖;減少內臟脂肪等各式各樣的作用。 關於乳鐵蛋白的安全性雖然目前並未具備充分的數據 ,亦有部分意見認爲孕婦應避免過量攝取,但由於其爲母 乳及牛乳中大Μ含有之成分,因此認爲對人體造成不良影 響的可能性極低。 進而有關乳鐵蛋白的種類,牛乳中所含者係牛乳鐵蛋 白,人體內所含者爲人乳鐵蛋白,雖爲不相同之種類,但 人類攝取牛乳鐵蛋白時,該人血液中之人乳鐵蛋白亦增加 。目前尙未明瞭其作用機轉。乳鐵蛋白對消化酵素而言具 有較高的耐受性而不易被分解,但認爲其係與藉由消化酵 素而被以較大塊狀分解之乳鐵蛋白’可生成具較強抗菌作 用之乳鐡蛋白素(lactoferricin)此一機轉有關。 201141515 該等乳鐵蛋白的硏究中,具體可舉出如下述的文獻。 專利文件1係有關治療痤瘡的文獻,於申請專利範圍 中有「一種含乳鐵蛋白所構成之乳漿蛋白質劃分之使用, 其係爲調製針對治療痤瘡之經口的組成物」之記載。然而 並無有關其作用機轉之詳細記載。 專利文件2、3、4、5係有關抗發炎、皮脂調節、抗菌 活性及治療/3 -防禦素(defensin )(抗菌胜肽)產生促進 所引起之痤瘡之文獻。專利文件3中,有「已知水解乳蛋 白質以往於改善毛髮及皮膚狀態之際使用。該物質可改善 乾燥皮膚皮脂的分佈,且對油性肌膚可降低過多皮脂生成 而可使用爲皮脂調節劑。」之記載。然而並無有關乳鐵蛋 白之記載。專利文件6係有關由乳鐵蛋白的抗氧化活性引 起的抗掉髮效果之文獻,雖有關於減輕頭皮表面產生的氧 化壓力對毛囊造成的不良影響之內容,但並無有關乳鐵蛋 白之抗雄激素作用之記載。專利文件7係有關乳鐵蛋白的 抗發炎活性,改善頻尿及攝護腺肥大之文獻,記載有可鎭 靜化壓迫尿路的發炎,及使排尿順暢之內容,但並無抗雄 激素作用之記載。非專利文件1提及乳鐵蛋白對皮脂之效 果,但針對其詳細內容並無具體的揭示。 因此,先前技術中,雖已揭示乳鐵蛋白之治療痤瘡( 經口)、脂漏症(塗布)、抗掉髮(塗布)、改善頻尿效 果(經口)、及水解乳蛋白質之藉由調節皮脂而可治療痤 瘡等內容’但自乳鐵蛋白作用機轉之觀點而言,並未存在 有提及乳鐵蛋白之抗雄激素作用之文獻。 201141515 專利文件 專利文件1 :特表2008-533134號公報 專利文件2 :特表2008-501772號公報 專利文件3 :特表2007-523137號公報 專利文件4 :特表2003 - 89629號公報 專利文件 5 : W02005-077349 專利文件6 :特開2007-22923號公報 專利文件7 :特開2 0 0 6 - 2 0 6 5 4 7號公報 非專利文件 非專利文件 1 : “Supporting healthy complex ion from the inside with Praventin” DMV international 【發明內容】 [發明欲解決之課題] 本發明係提供具有高抗雄激素作用,無副作用且安全 性亦優異的抗雄激素劑,以及以提供以該抗雄激素劑爲有 效成分之皮脂分泌抑制劑及養髮劑爲目的。 [解決課題之手段] 爲解決上述課題針對數種天然素材進行篩選,結果發 現存在於哺乳動物之乳汁、淚液、唾液、膽汁等分泌物中 之乳鐵蛋白,具有高抗雄激素作用,本發明遂至完成。 本發明係有關一種抗雄激素劑’其特徵係含有乳鐵蛋 201141515 白。 本發明係一種皮脂分泌抑制劑,其特徵係混合上述之 抗雄激素劑爲有效成分。 本發明係一種養髮劑’其特徵係混合上述之抗雄激素 劑爲有效成分。 本發明係一種飲食品’其特徵係含有以乳鐵蛋白爲有 效成分之抗雄激素劑。 [發明的效果] 本發明係可製爲含漱劑、吸入劑、***錠等製劑,另 外尙可製成口香糖、糖果、錠菓、可吸食果凍、餅乾、巧 克力等零食,冰凍果汁水、飲料等食品提供日常性的利用 及攝取’對於男性禿、粗髮等頭髮疾病、***肥大、前 列腺腫瘤等疾病、病因爲皮脂分泌過多的痤瘡、脂漏症等 皮膚疾病之預防或治療有效。進而本發明之抗雄激素劑其 效力極高,且無副作用兼具極佳之安全性。 【實施方式】 [實施例] [實施例1 ] 睾固酮5 -甲型還原酵素阻礙試驗 <試驗方法> L於0.1ml之3.OmM睾固酮溶液(以丙二醇溶解)中, 201141515 加入 0.5ml 之 5mM Tris-HCl緩衝溶液(ρΗ7·2 )。 2.其次,再加入〇」ml之6.7mM NADPH溶液(溶解於 5mM Tris-HCl緩衝溶液(pH7.2))及〇.〇5ml之檢體溶液 (溶解於50%乙醇),以37°C預備加熱後,加入0.1ml之酵 素液(S-9,Oriental Yeast)並進行1小時之孵育。 本試驗中乳鐵蛋白係使用來自牛之乳鐵蛋白而調製, 對照則使用沙巴棕。 3 .之後,加入3 ml之二氯甲烷使反應停止,並加入 〇.5ml之內部標準溶液(0.1 mg/ml之對羥苯甲酸正己酯) ,搖盪10分鐘,再以3 000rpm離心10分鐘。 4.取出二氯甲烷層後進行減壓乾燥。於乾燥物中加入 5ml之甲醇並使其溶解,製成試料溶液。以HPLc定量所得 之試料溶液中殘留的睪固酮。 測定條件係使用YMC A-302 (內徑4.6*150mm )管柱 ’管柱溫度爲40°C,流速lml/分鐘之條件,以甲醇/水( 65/35)溶離,以254nm檢測。另外,測定係使用內部標準 物質法進行,阻礙率(%)依下述式計算。
阻礙率(%) =(A-B) /(C-B) xlOO A :加入試驗液之後時睪固酮之量 B :對照組3 0分鐘時睪固酮之量(使用5 0%乙醇溶液 Φ代試驗液,進行反應時睪固酮之量) C :對照組0分鐘時睪固酮之量(於混合Tris-HCl緩衝 -10- 201141515 溶液、睪固酮、試驗液及酵素液之後,加入NADPH之前 ’加入二氯甲烷後使反應停止時睪固酮之量) <試驗結果> 將所得結果示於圖1。由圖1可明顯得知,作爲陽性對 照之沙巴棕於濃度lOOOppm時,顯示有約35%的阻礙活性 ’但並未發現乳鐵蛋白對睪固酮5 -甲型還原酵素的阻礙活 性。 [實施例2 ] 雄激素受器結合阻礙試驗 <試驗方法> 1 ·將具有雄激素依存性之小鼠乳癌細胞SC-3細胞, 使用經2%DCC處理且含有FCS之MEM培養基,以每孔l.〇x 104 cells/well/ΙΟΟμΙ的細胞密度,播於96孔的微孔細胞培 養盤中,並於3 7 °C,5 % C 0 2 - 9 5 % a i r下進行培養。 2· 24小時後,以添加了試驗試料及10_8M DHT之HMB 培養基(含0.1% BSA之Ham F12 + MEM培養基)取代原先 的培養基後,再進行48小時之培養。 本試驗中乳鐵蛋白係使用來自牛之乳鐵蛋白而調製, 陽性對照則使用安體疏通(spironolactone)。 3.之後,藉由MTT還原法,測定藍甲臢(blue formazan)之最大吸收光區之570nm之吸光度,評價細胞 增殖程度。 -11 - 201141515 爲矯正附著細胞的影f#,同時測定6 5 〇 n m吸光度,以 —種吸光度之差作爲藍甲腰之生成量。同時,爲觀察僅使 用試料對於SC-3細胞之影響,與前述試驗並行,於HMB培 養基中加入未添加D Η T之試料,並進行相同的培養及測定 。進而對於對照組,對於以未添加試料及DHT之ΗΜΒ培養 基進行培養之情形,以及以未添加試料,僅添加D Η Τ之 Η Μ Β培養基進行培養之情形,亦進行相同的測定。根據測 定結果,以下述式計算表示抗雄激素作用之結合阻礙率。 結合阻礙率(%)=〔 {(A-B)-(C-D)}/(A-B)〕χΙΟΟ A :添加DHT,未添加試料時之吸光度 B :未添加DHT,未添加試料時之吸光度 C :添加DHT,添加試料時之吸光度 D :未添加DHT,添加試料時之吸光度 <試驗結果> 將所得結果示於圖2。由圖2可明顯得知’作爲陽性對 照之安體疏通於濃度i〇'5m時,顯示有約70%的結合阻礙 活性。而於乳鐵蛋白發現有濃度依存的結合阻礙活性,於 濃度爲60ppm時顯示有約85%的阻礙活性。 [實施例3] 皮脂腺細胞培養試驗 -12- 201141515 <試驗方法> 1 .本試驗係使用倉鼠皮脂腺細胞培養試劑組KB-1 000 (KURABO公司製)進行。 將來自正常黃金倉鼠耳殼之皮脂腺細胞,以每孔5 · 0 X 1〇4 cells/well的細胞密度,播於24孔的微孔細胞培養盤中 。於試驗期間內,以含有濃度爲1x1 (Γ6Μ睪固酮之培養基 進行培養。 2.培養數日至形成全面的聚合細胞層後,將培養基改 換爲含有試驗試料之培養基。每日進行培養基更換,並於 約2周之培養期間持續進行。 本試驗中,乳鐵蛋白係使用來自牛之乳鐵蛋白而調製 ,對照則使用安體疏通。 3. 之後,使用皮脂合成測定試劑組 SE-3 00 1 ( KURABO公司製),於各孔中添加WST-8溶液,並於37°C 下孵育30分鐘。取上清液以45 0nm波長測定,並計測活細 胞數目。 4. 進而以油紅0染色,並於以顯微鏡觀察同時,以 100%異丙醇進行萃取,再以5 2 0nm波長測定,計測脂質合 成量。 藉由下述式進行矯正,並比較每一個細胞的脂質合成 量。
每一個細胞的脂質合成量之比較=吸光度B/吸光度A 吸光度A :細胞數測定値(45 0nm ) 吸光度B :脂質合成量測定値(520nm) -13- 201141515 <試驗結果> 將所得結果示於圖3及圖4。由圖3及圖4可明顯得知’ 作爲陽性對照之安體疏通於濃度1x10·5Μ時,於顯微鏡觀 察及吸光度測定,其皮脂合成量均較對照組減少。反之’ 乳鐵蛋白於顯微鏡觀察及吸光度測定時,發現有濃度依存 地抑制皮脂合成量,於濃度爲1 〇 P P m、2 0 p p m時可抑制皮 脂合成量至60%。 其次,使用乳鐵蛋白,以常用方法調製錠劑、口香糖 、糖果、巧克力、餅乾、可吸食果凍、錠菓、藥錠、冰淇 淋、冰凍果汁水、飮料等。以下揭示其配方。且並未因該 等例示而限制本發明品之範圍。 [實施例4] 根據下述配方調製錠劑。 D-甘露糖醇 42.6% 乳糖 42.6 結晶纖維素 8.5 羥丙基纖維素 4.3 乳鐵蛋白 _2.0 10 0.0% [實施例5] 根據下述配方調製口香糖。 -14- 201141515 基礎橡膠 2 0.0% 砂糖 54.7 葡萄糖 14.5 水飴 9.3 香料 0.5 乳鐵蛋白 1.0 10 0.0% [實施例6] 根據下述配方調製糖果。 砂糖 5 0.0% 水飴 3 1.4 檸檬酸 1 .0 香料 0.2 L-薄荷醇 1.0 乳鐵蛋白 2.0 水 14.4 10 0.0% [實施例7] 根據下述配方調製巧克力。 -15- 201141515 可可塊 2 0.0% 全脂乳粉 20.0 可可脂 17.0 粉糖 40.45 卵磷脂 0.45 香料 0. 1 乳鐵蛋白 2.0 10 0.0% [實施例8] 根據下述配方調製餅乾。 砂糖 3 1.7% 低筋麵粉 26.8 片栗粉 26.8 奶油 3.2 雞蛋 10.2 小蘇打粉 0.3 乳鐵蛋白 1.0 1 ο ο. ο % [實施例9] 根據下述配方調製可吸食果凍。 -16- 201141515 聚糊***溶液 39.0% 山梨糖醇水溶液 8.0 巴拉金糖水溶液 9.0 麥芽糖水溶液 20.0 海藻糖水溶液 11.0 明膠 10.0 酒石酸 1.0 乳鐵蛋白_2.0 10 0.0% [實施例10] 根據下述配方調製錠菓。 砂糖 4 6.65 % 葡萄糖 19.0 蔗糖脂肪酸酯 0.2 香料 0.15 乳鐵蛋白 30.0 水 4.0 10 0.0% [實施例1 1] 根據下述配方調製錠劑。 -17- 201141515 砂糖 蔗糖脂肪酸酯 乳鐵蛋白 水 3 5.8 5 % 0.15 60.0 4.0 10 0.0% [實施例12] 根據下述配方調製冰淇淋。 蛋黃 11.0% 砂糖 14.0 牛乳 35.5 鮮奶油 37.0 香草籽 0.5 乳鐵蛋白 2.0 10 0.0% [實施例13] 根據下述配方調製冰凍果汁水。 橘子果汁 16.0% 砂糖 3 1.0 乳鐵蛋白 3.0 水 50.0 1 ο ο. ο % -18- 201141515 [實施例14] 3 0.0% 13.34 0.1 0.04 0.1 2.0 54.42 10 0.0% 根據下述配方調製飲料。 橘子果汁 高果糖玉米糖漿 檸檬酸 維他命C 香料 乳鐵蛋白 水 [產業上之可利用性] 本發明係可將具有抗雄激素作用之乳鐵蛋白作爲有效 成分’利用於添加至商品群之添加素材。另外,其不但可 適用於糖果及口香糖等製品’亦可適用於新穎的健康功能 製品。 【圖式簡單說明】 [圖1]表示睪固酮5 -甲型還原酵素阻礙活性之柱狀圖。 [圖2]表示雄激素受器結合阻礙率之柱狀圖。 [圖3 ]表示皮脂腺細胞中皮脂合成量之圖。 [圖4 ]表示皮脂腺細胞以油紅〇染色(皮脂腺細胞培養 試驗結果)之顯微鏡照片圖。 -19-
Claims (1)
- 201141515 七、申請專利範圍: 1. 一種抗雄激素劑,其特徵係含有乳鐵蛋白。 2. —種皮脂分泌抑制劑,其特徵係混合如申請專利範 圍第1項之抗雄激素劑爲有效成分。 3 ·—種養髮劑’其特徵係混合如申請專利範圍第1項 之抗雄激素劑爲有效成分。 4.—種飲食品’其特徵係含有以乳鐵蛋白爲有效成分 之抗雄激素劑。 -20-
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JP2832517B2 (ja) * | 1994-12-09 | 1998-12-09 | 雪印乳業株式会社 | 大腸菌付着阻止剤 |
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HK1173085A1 (zh) | 2013-05-10 |
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