201138858 六、發明說明： 【韻^ s月所屬技φ好領域】 相關申請之參照 本申請案係基於已先於日本提申之特願2010-158104 號（申請日：2010年3月15日），而主張其優先權利益，並參 照其揭露内容整體，而加以引用於此。 技術領域 本發明係有關於一種以燕窩之酵素分解物為有效成分 之人類皮膚表皮細胞增殖促進劑。且，本發明並有關於一 種包含上述有效成分之燕窩之酵素分解物之皮膚外用組成 物及化妝用料。 t ]| 發明背景 肌膚乾燥、皺紋、肌膚鬆他、彈性不佳等肌膚問題， 尤其文到女ϋ重視與關心。該等肌膚問題通常被認為係由 環境因素、老化等熟齡化因素，甚至壓力等心理因素等所 導致。尤其’近年紫外線增加及㉙之泛料所伴隨之生 活環境之賴化等所導輯肌㈣成孩㈣之環境因素 可謂正逐漸增加。 皮膚，且織係由表皮、真皮、皮下組織所構成。表皮係 形成皮膚最外層之構成要素，已知基本上更包含角質層、 _層1㈣'基底層。表皮中，於最下層之基底層可 ***新生新的角質細胞(表皮細胞)，細胞則逐漸朝上方改變 功能與形態而曰益上昇’成為角質層之角質細胞後，最後 201138858 乃自皮膚表面剝落。人類之健康皮膚自基底層内表皮細胞 之刀裂至上昇成為角質細胞再由皮膚表面剝落，據稱需時 28天私度。上述自表皮細胞之***至細胞表面之細胞剝剝 離之一連串流程稱為代謝，上述代謝在表皮反覆進行，而 就皮膚進行新陳代謝以維持健康之皮膚。 然而’已知熟齡化將導致上述代謝之周期延緩，並咸 認代謝周期之延緩將導致皮膚表面殘留老廢角質等，而造 成肌膚暗沈及肌膚乾燥之狀態等。 因此’促進皮膚表皮細胞之增殖，而活化皮膚之新陳 代謝’並維持代謝周期，使皮膚表面常保健康，即可提昇 肌膚滋潤度、調理肌膚紋理，可謂將促成生活品質（QOL) 之提昇。 表皮（上皮）細胞成長因子（Epidermal growth factor(EGF))係已知存在包含人類在内之動物體内之上皮 細胞再生因子，發現自老鼠之頜下腺之萃取物而為蛋白質 之一種(mEGF(老鼠EGF))〇EGF就各種動物均各自不同，老 鼠EGF並不直接在人體展現細胞增殖促進效果。亦已發現 人類E G F，並已知E G F可對皮膚表面之細胞作用而促進細胞 之增殖。另，舉例言之，特開2009-234980號公報中，已揭 露人類隨著年齡增長，EGF之分泌量亦逐漸減少（Life Sciences vol. 31，pp679-683, 1982)，此乃肌膚老化現象之因 素之一之内容。因此，咸認對肌膚補充EGF，即可促進人 類表皮之細胞新生（增殖），而維持良好肌膚狀態。 燕窩係金絲燕以自己的唾液構成絲狀而築成之巢，已 201138858 幾乎不含脂質。燕窩迄 之原料等。近年，亦已 至於外用劑及化妝用料 知其成分包含多量之_蛋白質，而 今使用於中菜作為食材，亦為中藥 開發以燕窩為原料之健康食品等， 等外用用途之應用例則尚少為人知 特開2003-095961號公報中p 4b _ Λ ^ 已揭路使用燕窩之酵素分 解物之口服用之美肌促進劑。妙 ’、刀 …、而，其中僅揭露口服用途， 而未就外用用途之利用可能性竹^ 、 可揭露及暗示。又，复 效果係肌膚保水度之提昇，而眘 ’、 現潤澤之肌理細緻之浙 膚，並未就促進表皮之細胞增殖Μ持代謝周期，而維持、 提昇肌理細緻等肌膚狀態等用途作任何揭露 W09·屬中，已揭露包含燕^含水溶劑萃取 物，特別是熱水萃取物之化妝用料。該㈣㈣—㈣ 皮膚保養用途，係使用於外用用途者。然而，其中使用之 萃取物係就燕寫加以熱水萃取去除固形物後而成者，與就 燕窩整體予以酵素處理而直接使用之燕㈣素分解物戴然 不同D且’關於其效果’亦與基於表皮之細胞增殖之促進 之本發明效果不同。 特開2009-234980號公報即有關於包含EGF關聯物質與 神經胺酸之美容組成物，其中，已揭露利用燕窩作為神經 胺酸之補充來源之一。然而，其中，神經胺酸係用於活化 EGF關聯物質者，EGF關聯物質乃必要之成分，而燕窩並非 必要成分，僅與蛋及牛乳同為神經胺酸之供給來源而已。 I：發明内容1 本發明人此次發現了燕窩之酵素分解物具有類似人類 201138858 EGF之優良人類表皮細胞增殖促進效果。與人類無密切關 聯之金絲燕所製造之燕窩之酵素分解物竟展現類似人類 EGF之細胞增殖促進效果，乃出乎預料之驚人發現。且， 酵素分解物所展現之效果較燕窩之水萃取物更明顯，此亦 為驚人之結果。本發明即以該等發現為依據。 故而，本發明之目的在提供可有效率地生產且高安全 性較之優良人類皮膚表皮細胞增殖促進劑，以及含有該劑 之外用劑及化妝用料。 本發明之人類皮膚表皮細胞增殖促進劑係以燕窩之酵 素分解物作為有效成分。 依據本發明之一適當態樣，前述酵素分解物係蛋白酶 所致之分解物。 依據本發明之一較佳態樣，前述酵素分解物之分子量 為 70〜200,000。 依據本發明之一更佳態樣，前述酵素分解物之平均分 子量為300〜50,000。 依據本發明之另一態樣，可提供含有本發明之人類皮 膚表皮細胞增殖促進劑之皮膚外用組成物。該組成物宜使 用於皮膚保養，更宜使用於調理肌膚紋理，並改善肌膚或 皮膚狀態。 依據本發明之另一較佳態樣，本發明之皮膚外用組成 物係化妝用料。依據本發明之另一更佳態樣，本發明之皮 膚外用組成物係化妝水、乳液、護膚霜、保濕液、護脣膏、 美容液面膜或洗面乳。 6 201138858 依據本發明之另一態樣，可提供一種燕窩之酵素分解 物的用途，其係用於製造人類皮膚表皮細胞增殖促進劑 者。依據本發明之再另一態樣，可提供一種燕窩之酵素分 解物之用途，其係用於製造皮膚保養用之皮膚外用組成物 者。 依據本發明之又另一態樣，可提供一種人類皮膚表皮 細胞之增殖促進方法，包含：將有效量之本發明有效成分 之酵素分解物應用於人類皮膚的步驟。。依據本發明之又 再一態樣，可提供一種皮膚保養方法，包含：將有效量之 本發明有效成分應用於人類皮膚的步驟之。同樣地，可提 供一種調理肌膚紋理及提昇或改善肌膚狀態之方法，包 含：將有效量之本發明有效成分應用於人類皮膚的步驟之。 依據本發明，使用燕窩之酵素分解物作為有效成分， 即可較以往更顯著促進人類皮膚表皮細胞增殖。且，基於 上述人類皮膚表皮細胞增殖促進功能，將酵素分解物使用 於皮膚外賴途或化㈣料用途，即可維持皮膚之代謝周 期或恢復延緩之周期’而調理肌纽理及維持或改善肌膚 或皮膚狀態。如上所述，依據本發明，可實現消費者之q〇l 之提昇。X ’所使社酵素分解物係使用㈣知為食材之 燕高，故安全性較高’消費者接受度亦較高。進而，酵素 分解物可直接就燕窩進行酵錢理而取得，故可有效率地 加以製造’並降低製造成本。 圖式簡單說明 第1圖係顯示燕窩樣本中 之延酸量之測定結果之圖表 〇 201138858 第2圖係係顯示燕窩樣本中之蛋白質量之測定結果之 圖表。 第3圖係顯示GPC-HPLC之燕窩酵素分解物1之測定結 果之圖表。 第4圖係_示GPC-HPLC之燕窩酵素分解物2之測定結 果之圖表。 第5圖係顯示第（3)實施例之結果之圖表。 第6圖係顯示第（4)實施例之結果之圖表。 【貧施冷式】 發明之具體說明 人類皮膚表皮細胞增殖促進劑及有效成分 本發明之人類皮膚表皮細胞增殖促進劑一如前述，係 以燕窩之酵素分解物作為有效成分。 酵素分解物之原料燕窩係金絲燕將自己的唾液構成絲 狀而築成之巢，其成分包含多量之醣蛋白質，幾乎不含脂 質。在此，金絲燕係指雨燕科（Apodidae)之金絲燕屬 (Collocalia)下之燕類，可例舉諸如小灰腰金絲燕（Apodidae Collocalia francica)、苔巢金絲燕（Apodidae Collocalia salanga)、爪。圭金絲燕(Apodidae Collocalia fuciphaga)、印度 食用巢金絲燕(Apodidae Collocalia inexpectata)、褐腰金絲 燕（Apodidae Collocalia vestita)、白腹金絲燕（Apodidae Collocalia esculenta)、大金絲燕（Apodidae Collocalia maximus)等。 本發明中，燕窩可使用市售品。一般市售之燕窩從僅 201138858 去除羽毛及糞便等污垢而經清洗者，至收集燕窩之碎屑並 反覆進行漂白與清洗而成形者，涵括眾多種類，但本發明 所使用之燕窩則宜使用未在前置處理時經過度清洗及漂白 等之燕窩。 燕窩之酵素分解物係指就燕窩進行酵素反應加以分解 所知者，可使用蛋白酶或含有蛋白酶之複合酵素等分解燕 窩或其處理物而製得。在此，分解處理所使用之酵素宜為 蛋白酶’使用胰酶則更佳。 燕窩之酵素分解物通常可先經加熱殺菌處理，然後於 水中進行酵素處理。殺n處理之條件可適當參考慣用之加 熱殺菌條件而設定，酵素處理之條件亦可對應所使用之酵 素種類及燕窩狀態而適當加以設定。 舉更具體之例，則可依以下程序調製燕窩之酵素分解 物。 舉例言之，對已磨碎之燕离加人其質量之1G〜50倍之水 而使其此脹後，在的〜丨机下進行加熱殺菌處理$秒至％ 刀知。然後，直接添加適量之酵素，而在酵素之最佳阳、 最佳溫度下進行酵素反應G5〜48小時。酵素反應結束後， 藉加熱處理f㈣素不科化，並反舰而去除不溶 物’即可製得絲之酵素分解物q，上述酵素分解物亦 可視需要而乾燥祕狀。另，上述各程序中，亦可進行適 當pH調整、脫色、脫臭等操作。 另一例則在就燕寓之處理物進行酵素分解時，對粒徑 2mm以下、而以15G_以下更佳之已磨碎之燕窩加入其質 201138858 量之10〜100倍之水，並在^60。 〇c下予以靜置或攪拌〇5 小時而使其膨脹後’在6〇〜13代下進行加熱㈣處 〜30分鐘’並視需料予叫_得毅，魏歧相 同之酵素處理，而可製得燕k酵素分解物。相 酵素分解物之調製法係酵素反應之時間愈長， 之分解程度愈高，而可製得包含更低分子之蛋白質之酵^ 分解物。本發明中，酵素反應時間為〇5〜48小時*則W 小時為佳，3〜36小時更佳，約24小時程度則最佳，並宜使6 用業經較充分之酵素反應之酵素分解物。 如此製得之燕窩之酵素分解物之分子量宜為 70〜200,000，70〜180,000則較佳。分子量為7〇〜15〇 〇〇〇則更 佳，70〜120,000則最佳。 又，燕窩之酵素分解物之平均分子量（重量平均分子量) 宜為300〜100,000，300-70,000則較佳。平均分子量為 300〜50,000則更佳。 皮膚外用組成物 本發明之人類皮膚表皮細胞增殖促進劑雖可單獨直接 使用’但亦可内含於化妝用料、醫藥品、準藥物等各種皮 膚外用組成物中作為添加劑，而製得具有人類皮膚表皮細 胞之增殖促進效果之組成物^所製得之組成物可有效使用 於皮膚保養，更宜用於調理肌膚紋理，並改善或維持肌膚 或皮膚狀態。 本發明之有效成分之燕窩之酵素分解物具有使egf_r 表現皮膚構成細胞實際增殖之效果，該效果並較燕窩之酵 10 201138858 素分解物之未添加組及燕寫之熱水萃取物組更為顯著（後 述之第1實施例之測試結果）。且，本發明之有效成分之燕 窩之酵素分解物已展現實際磷酸化EGF受體(EGF-R)之效 果（後述之第2實施例之測試結果）。EGF-R之磷酸化係指燕 窩之酵素分解物實際上保有作為EGF-R之配體之作用， 即，代表實際上保有類EGF之作用。 因此，本發明中，人類皮膚表皮細胞增殖促進活性係 指可使人類皮膚表皮細胞實際增殖。燕窩之酵素分解物 中，被認為含有可作用一如EGF受體之配體之成分，結果 則推論可促進EGF活性，而促進表皮細胞之增殖。另，上 述純屬理論而非用於限定本發明者。 因此’依據本發明，使用燕窩之酵素分解物，即可期 待類EGF之效果，結果則可促進表皮之細胞增殖，而維持 表皮之代謝周期或恢復延緩之周期，而改善人類皮膚之新 陳代謝。因此，在調理肌膚紋理，改善及提昇肌膚或皮膚 狀態方面之效果明顯。且，亦可謂具備預防皮膚狀態之惡 化之效果。 另’本說明書中，所謂肌膚或皮膚之狀態之「改善或 維持」之用語’包含其狀態之提昇、調節、狀態惡化之延 緩或減輕等意義。 故而’一如前述，依據本發明之其它態樣，可提供構 成含有本發明之人類皮膚表皮細胞增殖促進劑之皮膚外用 組成物。皮膚外用組成物則宜提供作為醫藥品、準藥物或 化妝用料，而更宜提供作為化妝用料。 201138858 之 本發明之皮膚外用組成物（尤其化妝用料）中，有t成八 -燕窩酵素分解物之含量(乾物換算)並無特別之限制依2 成物开> 態而不同，但一般以固形物計算而為〇 〇〇 1 曰 =:而一5重量百分比之範圍為佳，可對= 物之φ態而適當加以變更。 或，本發明之皮膚外用組成物中燕窩酵素分解物之入 量(乾物換算)宜為可使每平方公分之皮膚每 3 〇._mg以上，而以〇 〇〇lmg〜㈣為佳之量。 達 在本發明之皮膚外用組成物包含有效成分， 類皮膚表皮細胞增殖促進效果之限度内，本發明之=人 可進而包含其它任意成分。上述之任意成分可為諸如^劑 外用劑及化妝用料所慣用之久、 為诸如皮膚 I員用之各種成分，可例舉諸 劑、低級醇、多價醇、 保减 ⑭醣類、界面活性劑、緩衝劑 劑、女疋劑、增黏劑 礼化 PH調整劑、香料、色素防腐抗菌劑、螫合劑、 維他命類、絲、卜収M1 t外線散射劑、 類、植物萃取類、抗氣私㈤水月生月太糖醇類、酵素 珍珠粉等。^化物質類、滑石粉、黏土、花粉、 “此:濕劑可例舉諸如聚乙二醇、聚丙二醇、内 醇、丙一醇、山梨糖醇 丙二 醣類、胺錢、料械軟#料酸性點多 ^/蛋白、膠原蛋白胜肽、彈力蛋白笼 低級醇可例舉諸I白荨。 多價醇可例舉諸=㈣、異丙醇等。 乙二醇、丙二醇、h 異戊㈣、雙異戊四醇、 聚兩二醇、1，3-丁二醇等。 12 201138858 醣類則可例舉諸如葡萄糖、半乳糖、麥芽糖、乳糖、 木糖、D-葡萄醣醛酸、蔗糖、葡萄糖胺、半乳糖胺、〇_山 梨醣醇、山梨醇酐、纖維素、澱粉等醣類、寡糖類、多醣 類及其等之誘導體等。 本發明之皮膚外用組成物可採用公知之方法適當調配 包含燕窩酵素分解物之人類皮膚表皮細胞增殖促進劑及上 述之任意成分，而構成化妝水、乳液、護膚霜（諸如除紋霜、 滋養霜、塑身霜）、保濕液、敷面劑（諸如美容液面膜）、化 妝水、洗面乳、身體乳、身體霜等各種化妝用料之慣用產 品形態。本發明之皮膚外用組成物亦可進而提供作為粉底 類、護脣膏、脣膏、眼影、腮紅等彩妝化妝品及防曬用產 〇〇防臭化妝品等藥用化妝品、洗髮精及调絲精、造型品 等髮妝品、皮膚清潔劑及入浴劑之浴用化妝品等之使用用 途。 依據本發明之另一態樣，一如前述，可提供對人類皮 膚應用本發明之有效成分之酵素分解物之有效量之人類皮 膚表皮細胞之增殖促進方法。另，在此所謂「有效量」係 指因使用而可於體内之預定領域内發揮人類皮膚表皮細胞 增殖促進活性之充足量。 進而，依據本發明之又另一態樣，一如前述，可提供 對人類皮膚應用本發明之有效成分之有效量之皮膚保養方 法。同樣地，亦可提供對人類皮膚應用本發明之有效成分 之有致量之調理肌膚紋理之肌膚狀態提昇或改善方法。 另’在此所謂「有效量」係指可獲致預期效果之充足量。 13 201138858 另，本說明書中，使用「約」及「程度」之用語之值， 包含在達成設定其值之目的之前提下，本業之技術人員所 可容許之值之變動之意義。舉例言之，意指可容許預定值 或範圍之20%以内之’聋動，而以1 〇%以内為佳，5%以内則 更佳。 實施例 參照以下之例具體說明本發明，但本發明並不受限於 以下之例。 (1)燕窩樣本之調製 (i_i)燕窩酵素分解物1 藉碾磨機磨碎卞售之未漂白之燕窩，而製得100篩目大 小(粒徑15 0 μ m以下）之燕窩粉末，再對該燕窩粉末加入約5 〇 倍量（質量）之水而在5 C下使其膨脹2〇小時後，在丨2i〇c下進 行加熱殺菌處理15分鐘。 在製得之處理液冷部後，調整pH，並對燕窩粉末添加2 質量百分比之量之含蛋白酶酵素(商品名「pa_atinF， 天野製藥株式會社出品），而在价下進行反應3小時。 將上述酵素反應液調整成pm.〇後，在贼下加執$分 鐘而使酵素不活性化’再加以輯相收舰(水溶性部 刀）。濃縮上述親後東結乾燥而祕「燕窩酵 解物1」 〇-2)燕窩酵素分解物 除酵素反應之時間改為24小日#,、，w t 4小時以外，製程均與前述（iq) 之酵素分解物i相同，而製得「_酵素分解物2」。 14 201138858 (1-3)燕窩熱水萃取物（比較例） 藉碾磨機磨碎市售之未漂白之燕窩（乾燥物）’並將其磨 碎物加入l〇〇〇ml之蒸館水中，再藉加熱迴流進行2小時之萃 取處理。加熱迴流後，再予靜置，然後分離取得上澄液加 以過濾，而回收濾液。殘渣則進而與前述相同，予以加入 1000ml之蒸餾水中，藉加熱迴流而進行再萃取處理後，過 濾固形物殘渣而加以去除，並回收濾液。所得之濾液（萃取 液）則一同在減壓環境下加以濃縮，所得之濃縮物則予以;;東 結乾燥，而製得黃白色之「燕窩熱水萃取物」（比較例 (2)燕窩樣本之分析 製得之各燕窩樣本（燕窩酵素分解物1、燕窩酵素分解 物2及燕窩熱水萃取物）中之涎酸量與蛋白質量已測定如 下。 涎酸之量係在各試料之酸性水解後藉高速液體層析法 而測定游離N-乙醯神經胺酸所得。 蛋白質之量則使用基於Brad ford法之Bi〇 Ra(j公司之 Protein Assay Kit 而力σ 以測定。 結果則如第1及第2圖所示。 其次，為測定燕窩酵素分解物1及燕窩酵素分解物2所 含蛋白質之分子量及其分布，而在以下條件下，進行了 Gpc 之分析。 [HPLC之測定條件] •管柱：Sh〇deXAsahipakGS320HQ(v7.6x3〇〇_)201138858 VI. Description of the invention: [Rhyme ^ s month belongs to technology φ good field] Related application reference This application is based on the special request 2010-158104 (application date: March 15, 2010) that has been submitted before Japan. And claim its priority interests, and refer to its disclosure of the content as a whole, and cited here. BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a human skin epidermal cell proliferation promoting agent comprising an enzyme decomposition product of bird's nest as an active ingredient. Further, the present invention relates to a skin external composition and a cosmetic material for an enzyme decomposition product of bird's nest containing the above-mentioned active ingredient. t ]| BACKGROUND OF THE INVENTION Skin problems such as dry skin, wrinkles, loose skin, and poor elasticity are particularly important to the attention and concern of the son-in-law. These skin problems are usually thought to be caused by environmental factors, ageing factors such as aging, and even psychological factors such as stress. In particular, the environmental factors such as the increase in ultraviolet rays in recent years and the living environment accompanying the general publicity of 29 (4), the environmental factors of children (4) are gradually increasing. The skin, and the woven system is composed of epidermis, dermis, and subcutaneous tissue. The epidermis forms a constituent element of the outermost layer of the skin, and is known to substantially further comprise a stratum corneum, a _layer 1 (four) 'base layer. In the epidermis, new keratinocytes (epidermal cells) can be disrupted in the basal layer of the lowermost layer, and the cells gradually change function and morphology upwards and increase the keratinocytes of the stratum corneum. Finally, 201138858 peels off from the surface of the skin. . Human healthy skin from the cleavage of the epidermal cells in the basal layer to the rise of keratinocytes and then peeling off the surface of the skin, said to take 28 days of private. A series of processes described above for the separation of cells from the epidermal cells to the cell surface are called metabolism, and the above metabolism is repeated in the epidermis, and the skin is metabolized to maintain healthy skin. However, it is known that aging is expected to cause the above-mentioned metabolic cycle to be delayed, and the delay in the metabolic cycle will cause the skin to leave old dead skin cells and the like, resulting in dull skin and dry skin. Therefore, 'promoting the proliferation of skin epidermal cells, and activating the metabolism of the skin' and maintaining the metabolic cycle, so that the skin surface is always healthy, can improve the skin's moisture level and regulate the skin texture, which can promote the improvement of quality of life (QOL). Epidermal growth factor (EGF) is a kind of protein known as the epithelial cell regeneration factor in animals including humans. It is a kind of protein found in the submandibular gland of mice (mEGF (mouse) EGF)) EGF is different for each animal, and mouse EGF does not directly exhibit cell proliferation promoting effects in humans. Human E G F has also been found, and E G F is known to act on cells on the surface of the skin to promote cell proliferation. In addition, for example, Japanese Laid-Open Patent Publication No. 2009-234980 discloses that human secretion increases with age, and the secretion of EGF is gradually reduced (Life Sciences vol. 31, pp 679-683, 1982), which is a skin aging phenomenon. The content of one of the factors. Therefore, the addition of EGF to the skin promotes cell renewal (proliferation) of the human epidermis and maintains a good skin condition. The bird's nest swiftlets are made of silk by their own saliva, and they have almost no lipids in 201138858. The raw materials of the bird's nest are as follows. In recent years, it has been known that the ingredients for external use and cosmetic materials contain a large amount of protein. Today, it is used in Chinese food as a food ingredient, and it is also used to develop health foods such as bird's nest as a raw material for Chinese medicine, and there are still few applications for external use. It is known that P 4b _ Λ ^ in the publication of Japanese Patent Publication No. 2003-095961 has been used for the oral use of the skin growth promoter of the enzyme decomposition product of bird's nest. Wonderful, knives, and, of course, only expose oral use, and the possibility of use for external use is not disclosed, can be disclosed and implied. In addition, the complex effect is the improvement of the skin's water retention, and the careful and delicate skin texture of the skin does not promote the metabolic cycle of the cells of the epidermis, but maintains, enhances the skin texture and other skin conditions. It is disclosed in the genus W09 that a cosmetic material containing an aqueous solvent extract, particularly a hot water extract, has been disclosed. The (4)(4)-(4) skin care application is for use in external use. However, the extract used therein is obtained by extracting and removing hot solids by hot water extraction, and the Yan (tetra) decomposition product directly used for the enzyme treatment of the bird's nest is different from D and 'about its effect' The effects of the present invention differ from the promotion of cell proliferation based on epidermis. Japanese Laid-Open Patent Publication No. 2009-234980 is directed to a cosmetic composition comprising an EGF-related substance and a tranexamic acid, wherein the use of bird's nest as a supplemental source of ceramide is disclosed. However, in the case where the ceramide is used to activate the EGF-related substance, the EGF-related substance is an essential component, and the bird's nest is not an essential component, and is only a source of supply of ceramide for the egg and the milk. I: SUMMARY OF THE INVENTION The present inventors have found that the enzyme decomposition product of bird's nest has an excellent human epidermal cell proliferation promoting effect similar to human 201138858 EGF. The enzyme decomposition of bird's nest made by the swiftlet, which is not closely related to human beings, shows a cell proliferation-promoting effect similar to human EGF, which is unexpectedly unexpected. Moreover, the effect of the enzyme decomposition product is more obvious than that of the bird's nest water extract, which is also an amazing result. The present invention is based on these findings. Accordingly, an object of the present invention is to provide an excellent human skin epidermal cell proliferation promoting agent which can be efficiently produced and has high safety, and an external preparation containing the same and a cosmetic preparation. The human skin epidermal cell proliferation promoting agent of the present invention contains an enzyme decomposition product of bird's nest as an active ingredient. According to an appropriate aspect of the present invention, the aforementioned enzyme decomposition product is a decomposition product by protease. According to a preferred embodiment of the present invention, the aforementioned enzyme decomposition product has a molecular weight of 70 to 200,000. According to a preferred aspect of the present invention, the aforementioned enzyme decomposition product has an average molecular weight of from 300 to 50,000. According to another aspect of the present invention, a skin external composition containing the human skin epidermal cell proliferation promoting agent of the present invention can be provided. The composition should be used for skin care, and it is more suitable for conditioning skin texture and improving skin or skin condition. According to another preferred aspect of the present invention, the external composition for skin of the present invention is a cosmetic material. According to another more preferred aspect of the present invention, the external composition for skin of the present invention is a lotion, lotion, skin cream, moisturizer, lip balm, cosmetic liquid mask or facial cleanser. 6 201138858 According to another aspect of the present invention, there is provided a use of an enzyme decomposition product of bird's nest for the production of a human skin epidermal cell proliferation promoter. According to still another aspect of the present invention, there is provided a use of an enzyme decomposition product of bird's nest for use in the manufacture of a skin external composition for skin care. According to still another aspect of the present invention, a method for promoting proliferation of human skin epidermal cells comprising the step of applying an effective amount of an enzyme decomposition product of the active ingredient of the present invention to human skin can be provided. . According to still another aspect of the present invention, there is provided a skin care method comprising the step of applying an effective amount of the active ingredient of the present invention to human skin. Similarly, a method of conditioning the skin and enhancing or improving the condition of the skin can be provided, comprising the step of applying an effective amount of the active ingredient of the present invention to human skin. According to the present invention, the use of the enzyme decomposition product of bird's nest as an active ingredient can significantly promote the proliferation of human skin epidermal cells more than before. Further, based on the above-mentioned human skin epidermal cell proliferation promoting function, the enzyme decomposition product can be used for the purpose of maintaining the skin's metabolic cycle or restoring the cycle of delaying the skin's metabolic cycle or restoring the period of the skin's metabolism and maintaining or improving. Skin or skin condition. As described above, according to the present invention, the improvement of the consumer can be achieved. X ’ uses the enzyme decomposition products (4) to know that the ingredients are high, so the safety is higher ‘the consumer acceptance is also higher. Further, the enzyme decomposition product can be directly obtained from the fermentation of the bird's nest, so that it can be efficiently manufactured and the manufacturing cost can be reduced. BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is a graph showing the results of measurement of the amount of acid in the bird's nest sample. 〇 201138858 Fig. 2 is a graph showing the measurement results of the amount of protein in the bird's nest sample. Fig. 3 is a graph showing the results of measurement of the bird's nest enzyme decomposition product 1 by GPC-HPLC. Fig. 4 is a graph showing the results of measurement of bird's nest enzyme decomposition product 2 by GPC-HPLC. Fig. 5 is a graph showing the results of the (3) embodiment. Fig. 6 is a graph showing the results of the (4) embodiment. [Effective Example of the Invention] The human skin epidermal cell proliferation promoting agent and the active ingredient of the human skin epidermal cell proliferation promoting agent of the present invention are as described above, and the enzyme decomposition product of bird's nest is used as an active ingredient. The raw material of the enzyme decomposition product, the swallowlet swiftlet, forms a nest of silk into its own saliva, and its composition contains a large amount of glycoprotein, which is almost free of lipids. Here, the swiftlet refers to the swallows under the genus Collocalia of Apodidae, such as Apodidae Collocalia francica and Apodidae Collocalia. Salanga), claws. Apodidae Collocalia fuciphaga, Apodidae Collocalia inexpectata, Apodidae Collocalia vestita, Apodidae Collocalia esculenta, Apodidae Collocalia maximus) and so on. In the present invention, commercially available products can be used for the bird's nest. Generally, the bird's nest, which is commercially available, removes the dirt such as feathers and feces from 201138858, and collects the debris of the bird's nest and repeatedly bleaches and cleans it. It includes many types, but the bird's nest used in the present invention should be used. Bird's nest that has not been cleaned and bleached before pretreatment. The enzyme decomposition product of bird's nest refers to a compound which is decomposed by an enzyme reaction in bird's nest, and can be obtained by decomposing a bird's nest or a treatment thereof using a protease or a compound enzyme containing a protease. Here, the enzyme used in the decomposition treatment is preferably a protease, and it is more preferable to use trypsin. The enzyme decomposition products of bird's nest can usually be heat-sterilized and then treated with enzymes in water. The conditions for killing the n treatment can be appropriately set with reference to the conventional heating and sterilization conditions, and the conditions for the enzyme treatment can be appropriately set depending on the type of the enzyme to be used and the state of the bird's nest. To be more specific, the enzyme decomposition of bird's nest can be modulated according to the following procedure. For example, after the pulverized swallow is added to the mass of 1G to 50 times of its mass to make it swell, the sterilizing treatment is carried out under the 丨 machine for $second to %. Then, directly add the right amount of enzyme, and carry out the enzyme reaction G5~48 hours at the best yang and optimal temperature of the enzyme. After the completion of the enzyme reaction, the f(tetracycline) is not treated by heat treatment, and the insoluble matter is removed by anti-shipping, and the enzyme decomposition product q of the silk can be obtained, and the above-mentioned enzyme decomposition product can also be dried and secreted as needed. Further, in the above respective procedures, an appropriate pH adjustment, decolorization, deodorization, and the like can be performed. In another case, when the enzyme is decomposed in the treatment of Yan Yu, the water is less than 2 mm, and the ground bird's nest with a purity of 15 G_ or less is added to the water of 10 to 100 times the amount of 201138858, and at 60. . 〇c, let stand or stir for 5 hours and let it swell, then heat it at 4 〇~13 generations (four) ~ 30 minutes' and treat it as _ Deyi, Wei Qi, the same enzyme treatment, and It can produce yan k enzyme decomposition products. The longer the reaction time of the enzyme decomposition product, the higher the decomposition degree, and the yeast decomposition product containing the lower molecular protein can be obtained. In the present invention, the reaction time of the enzyme is 〇5 to 48 hours*, preferably W hours, more preferably 3 to 36 hours, and most preferably about 24 hours, and it is preferable to use 6 enzymes which are reacted with a sufficient enzyme reaction. . The enzyme decomposition product of the bird's nest thus obtained preferably has a molecular weight of 70 to 200,000, preferably 70 to 180,000. The molecular weight is 7 〇 15 15 〇 更 is better, and 70 〜 120,000 is the best. Further, the average molecular weight (weight average molecular weight) of the enzyme decomposition product of the bird's nest is preferably from 300 to 100,000, and preferably from 300 to 70,000. More preferably, the average molecular weight is from 300 to 50,000. Skin external composition The human skin epidermal cell proliferation promoter of the present invention can be used as it is, but it can also be contained in various skin external preparations such as cosmetic materials, pharmaceuticals, and quasi drugs. The composition of the skin epidermal cell proliferation promoting effect can be effectively used for skin care, and is more suitable for conditioning skin texture and improving or maintaining skin or skin condition. The enzyme decomposition product of the bird's nest of the active ingredient of the present invention has the effect that the eggf_r expresses the actual proliferation of the skin-constituting cells, and the effect is more than the un-added group of the bird's nest leaver 10 201138858 and the hot-water extract group written by Yan. Significant (test results of the first embodiment described later). Further, the enzyme decomposition product of the bird's nest of the active ingredient of the present invention has exhibited the effect of actually phosphorylating the EGF receptor (EGF-R) (test results of the second embodiment described later). Phosphorylation of EGF-R means that the enzyme decomposition product of the bird's nest actually retains the function as a ligand for EGF-R, i.e., represents the effect of actually retaining EGF-like. Therefore, in the present invention, human skin epidermal cell proliferation promoting activity means that human skin epidermal cells can be actually proliferated. In the enzyme decomposition product of bird's nest, it is considered to contain a component which acts as a ligand for the EGF receptor, and as a result, it is inferred that it promotes the activity of EGF and promotes the proliferation of epidermal cells. In addition, the above is purely a theory and is not intended to limit the inventors. Therefore, according to the present invention, the enzyme decomposition product of bird's nest can be used to treat the effect of EGF-like, and as a result, the cell proliferation of the epidermis can be promoted, and the metabolic cycle of the epidermis or the period of recovery delay can be maintained, and the metabolism of human skin can be improved. Therefore, the effect of conditioning the skin texture, improving and improving the skin or skin condition is remarkable. Moreover, it can also be said to have the effect of preventing the deterioration of the skin state. In the present specification, the term "improvement or maintenance" of the state of the skin or skin includes the promotion or adjustment of the state, and the delay or reduction of the deterioration of the state. Therefore, as described above, according to another aspect of the present invention, a skin external composition comprising the human skin epidermal cell proliferation promoting agent of the present invention can be provided. The external composition for skin is preferably provided as a pharmaceutical, quasi-drug or cosmetic material, and is preferably provided as a cosmetic material. In the external composition for skin of the present invention (especially, a cosmetic material) of the present invention, there is no particular limitation on the content of the t-occupant-degraded enzyme decomposition product (dry matter conversion), but it is generally different depending on the state of the product. The solid content is calculated as 〇〇〇1 曰 =: and a range of 5 weight percent is preferable, and can be appropriately changed for the φ state of the object. Alternatively, the amount of the decomposition product of the bird's nest enzyme in the external composition for skin of the present invention (dry matter conversion) is preferably such that the skin per square centimeter is more than 3 _._mg, and preferably 〇1 mg to (4). In the case where the external composition for skin of the present invention contains an active ingredient and a skin-derived epidermal cell proliferation promoting effect, the present invention may further contain other optional components. Any of the above-mentioned components may be used for a long time such as a topical preparation for external use and a cosmetic preparation, and may be various components such as a skin I member, and may be exemplified by various agents, lower alcohols, polyvalent alcohols, and 14 sugars, and interfaces. Active agent, buffer agent, licking agent, viscosity increasing agent, pH adjusting agent, perfume, pigment antiseptic antibacterial agent, chelating agent, vitamins, silk, M1 t external scatter agent, class, plant extract, anti- Gas private (five) water month, month, too sugar alcohol, enzyme pearl powder. Chemical substances, talcum powder, clay, pollen, "This: the wet agent can be exemplified by polyethylene glycol, polypropylene glycol, internal alcohol, propanol, sorbitol, malonose, amine money, soft materials. #料酸点多^/protein, collagen peptide, elastin cage lower alcohol can be exemplified by I. The polyvalent alcohol can be exemplified by = (tetra), isopropanol, etc. ethylene glycol, propylene glycol, h different Ethylene (tetra), diisopentaerythritol, polybisdiol, 1,3-butanediol, etc. 12 201138858 Examples of sugars such as glucose, galactose, maltose, lactose, xylose, D-glucuronic acid, An saccharide such as sucrose, glucosamine, galactosamine, guanidine sorbitol, sorbitol anhydride, cellulose, starch, oligosaccharides, polysaccharides, and the like, etc. The skin external composition of the present invention can be used. A well-known method suitably mixes a human skin epidermal cell proliferation promoting agent containing a bird's nest enzyme decomposition product and any of the above components to form a lotion, an emulsion, a skin cream (such as a cream, a nourishing cream, a body cream), a moisturizing liquid, and a compressing agent. Mask (such as beauty liquid mask), lotion, face wash A conventional product form of various cosmetic materials such as milk, body lotion, body cream, etc. The skin external composition of the present invention can further provide makeup cosmetics such as foundations, lip balms, lipsticks, eye shadows, and blushes, and sunscreen preparations. Use of a medicinal cosmetic such as a deodorant cosmetic, a shampoo, a hair styling, a hair styling product, a skin cleansing agent, and a bathing agent for a bathing agent, etc. According to another aspect of the present invention, as described above, An effective amount of a method for promoting proliferation of human skin epidermal cells by applying an enzyme decomposition product of an active ingredient of the present invention to human skin. The term "effective amount" as used herein means that it can be used in a predetermined field in the body by use. A sufficient amount of human skin epidermal cell proliferation promoting activity. Further, according to still another aspect of the present invention, as described above, an effective amount of skin care method for applying the active ingredient of the present invention to human skin can be provided. Similarly, it is also possible to provide a method for enhancing or improving the condition of the skin which is applied to human skin using the effective ingredients of the present invention to condition the skin texture. The term "effective amount" as used herein refers to a sufficient amount to achieve the desired effect. 13 201138858 In addition, the meanings of the terms "approximately" and "degree" are used in this manual to include the meaning of changes in the values that can be tolerated by those skilled in the art before the purpose of setting the value. For example, it means that the predetermined value or range is within 20% of the range, and preferably within 1%, preferably within 5%. EXAMPLES The present invention will be specifically described with reference to the following examples, but the present invention is not limited by the following examples. (1) Preparation of bird's nest sample (i_i) Bird's nest enzyme decomposition product 1 Grinding the unbleached bird's nest by grinding mill to obtain a bird's nest powder of 100 mesh size (particle size below 15 μm), To the bird's nest powder, about 5 times (mass) of water was added and it was expanded at 5 C for 2 hours, and then heat-sterilized at 丨2i〇c for 15 minutes. After the obtained liquid cold portion of the treatment liquid, the pH was adjusted, and 2% by mass of the protease-containing enzyme (trade name "pa_atinF, produced by Amano Pharmaceutical Co., Ltd.") was added to the bird's nest powder, and the reaction was carried out for 3 hours at a price. After the above enzyme reaction solution is adjusted to pm. ,, the thief is added for $ minutes to make the enzyme inactive, and then the collection is completed (water-soluble knife). Concentrate the above-mentioned pro-Eastern knot dry and secret "Swallow Nest Fermentation Dissolve 1" 〇-2) The time of the enzyme decomposition of the bird's nest enzyme is changed to 24 small days #,,, wt 4 hours, the process is the same as the enzyme decomposition product i of the above (iq), and the result is "_ Enzyme Decomposition 2". 14 201138858 (1-3) Hot water extract of bird's nest (Comparative example) Grinding commercially available unbleached bird's nest (dry matter) by a mill and adding the ground material to the steaming water of l〇〇〇ml Then, it was subjected to extraction treatment by heating under reflux for 2 hours. After heating to reflux, it was allowed to stand still, and then separated to obtain a supernatant liquid and filtered, and the filtrate was recovered. Further, the residue was added to 1000 ml of distilled water in the same manner as described above, and after re-extraction treatment by heating under reflux, the solid residue was filtered and removed, and the filtrate was recovered. The obtained filtrate (extract solution) is concentrated together under reduced pressure, and the obtained concentrate is obtained;; the east knot is dried to obtain a yellow-white "swallow nest hot water extract" (Comparative Example (2) bird's nest sample The amount of tannic acid and the amount of protein in each bird's nest sample (skin nest enzyme decomposition product 1, bird's nest enzyme decomposition product 2 and bird's nest hot water extract) were determined as follows. The amount of tannic acid was acid hydrolysis in each sample. After the high-speed liquid chromatography method was used to determine the free N-acetyl ceramide, the amount of protein was determined by the Brad ford method of Bi〇Ra (J company's Protein Assay Kit and force σ to determine. The result is as the first And Fig. 2 Next, in order to measure the molecular weight and distribution of the protein contained in the bird's nest enzyme decomposition product 1 and the bird's nest enzyme decomposition product 2, Gpc analysis was carried out under the following conditions. [HPLC measurement conditions] Column: Sh〇deXAsahipakGS320HQ (v7.6x3〇〇_)
•管柱溫度：35°C 15 201138858 •移動相：50mM CH3COONH3 pH6.7 •流速：〇.4ml/min • s式料注入量：1〇μ1 •偵測器：紫外分光偵測器（以UV220nm測定） [使用之分子量標示器] • BSAMW66,000 •甘胺醯甘胺酸(Gly-gly)MW132.1 -三胜肽（Gly-gly-gly)MW189.17 .L-天冬胺酸(L-Asparaginic acid)MW133.1 • DL-天冬胺酸(DL-Asparaginic acid)MW150.1 • N-乙醯神經胺酸(N-Acetylneuraminic acid)MW309.3 測定結果則如第3及第4圖所示。 進而，已就燕窩酵素分解物所含蛋白質之分子量，使 用西方墨點法而加以評價。具體而言，係先依循前述之測 定法而確認燕窩酵素分解物中蛋白質之量。就來自燕窩酵 素分解物之蛋白質（1 〇〇μβ)使用SDS-PAGE而藉10〜15%之聚 丙醢胺加以分離。將已分散於膠體中之蛋白質轉印至 lmmunoblot PVDF 膜（聚偏二氣乙稀（p〇lyvinylidene difluoride)，取自Bio-Lad Lab公司）。再於包含3%之脫脂乳 與0.1%之Tween-20之PBS中對PVDF膜進行阻隔1小時，再 進行視覺上之判定。 另’評價蛋白質之分子量時，亦已參考公知文獻(Guo, C. T., Suzuki Y., et al.5 Antiviral Res. 70, pp. 140-146(2006)) 所揭露之資料。 16 201138858 由上述結果可知，燕离酵素分解物中所含之蛋白質分 子量存在約7G〜12G，GGG程度（另，視評價條件之不同，上限 值可達200,000程度）之範圍内。 進而，由所得結果可知，燕窩酵素分解物丨之蛋白質之 平均分子量約為55，咖。又，可知燕窩酵素分解物2之蛋白 質之平均分子量約為33,000。 (3)酵素處理燕窩之細胞增殖促進活性 已使用E G F - R表現皮膚構成細胞檢討酵素處理燕寓之 類EGF作用，即，細胞增殖促進活性。 ⑴材料： 細胞係使用自ATCC購入之來自人類表皮（皮膚）之 EGF-R表現細胞A431細胞。 燕窩酵素分解物則使用由上述（1)製得之燕萬酵素分解 物1(酵素處理3小時）、燕窩酵素分解物2(酵素處理24小 時）、燕窩熱水萃取物。 (ii)實驗方法 藉已添加3%FCS之DME培養基對已調成5xl04cells/ml 之A431細胞按以下之各種濃度添加燕窩酵素分解物或燕窩 熱水萃取物，並在96孔平底盤中按5><103ceUs/lml/well之條 件進行培養。 最終添加濃度： .燕窩熱水萃取物(heart) : 1% ’ 〇·1%，0.01% •燕窩熱水萃取物 l(Enzl) : 1% ’ 0.1%，0.01% •燕窩熱水萃取物2(Enz2) : 1%，0.1%，0.01% 17 201138858 又，亦準備了未添加燕窩酵素分解物及r_EGF2未添加 組進行培養以作對照之用。 、、:田胞之增殖則長期採用Cell Counting Kit-8(同仁化學 研究所公司出品）(3H胸腺嘧啶之嵌入測試之替代法)加以檢 討’並調查類EGF作用之有無。 結果’以ABS(吸光度）代表吸收度在46〇nm附近為極大 之水溶性曱臢之色素量。 結果即如第5圖所示。 由結果可知，與未添加控制組(A431/3%FCS)相較，已 明確分為細胞增殖抑制組與增殖組。 燕寓酵素分解物1及燕窩酵素分解物2之添加組（尤其 0.1%及0.01%之濃度）均已確認明顯之增殖促進效果。 而，燕窩熱水萃取物之1%添加組即便與未添加控制組 比較，亦已極端抑制細胞增殖。燕窩熱水萃取物之〇1%添 加組及0.01添加組中，細胞增殖均低於未添加控制組。 因此’已確認燕窩酵素分解物可促進細胞增殖，而燕 窩熱水萃取物則反而抑制或妨礙細胞增殖之傾向。 (4)燕窩酵素分解物之類EGF作用之驗證(EGF_R_复化之檢討) 已以EGF-R表現皮膚構成細胞之egf受體磷酸化為指 標而檢討燕窩酵素分解物之類EGF作用。 ⑴材料： 細胞係使用自ATCC購入之來自人類表皮（皮膚）之 EGF-R表現細胞A431細胞。 燕窩酵素分解物則使用由上述(1)製得之燕窩酵素分解 18 201138858 物1(酵素處理3小時）、燕窩酵素分解物2(酵素處理24小時）。 已使用人類重組型EGF(r-EGF)(取自Protein Express)作 為正控制。 EGF-R之填酸化則使用Active EGF receptor EIA套件 (Takara公司）進行測定。 (ii)細胞培養： 藉已添加3%FCS之DME培養基分別對已調成 5 x 1 〇4cells/ml之A431細胞按以下之各種濃度添加燕窩酵素 分解物與作為正控制之r-EGF，並於24孔平底盤中依 104cells/lml/well之條件進行培養。 最終添加濃度： . •燕窩酵素分解物l(Enzl) ·· 0.1%，〇.〇1〇/0 • •燕窩酵素分解物2(Enz2) : 0.1%，0.01% • r-EGF : 〇.15ng/ml 又’亦準備了未添加燕窩酵素分解物及r-EGF之未添加 級進行培養以作對照之用。 (iv)檢體之調整及測定： 培養10小時至50小時（資料為1〇〜34小時)之期間内，長 期地回收細胞，並使用測定套件所附之受體萃取用緩衝液 6周整了檢體。去除培養上澄液後，依5〇|al/well加入受體萃 取用緩衝液，並使用細胞刮构（Ceii scraper)刮取細胞，而將 2Well份之細胞萃取液積存於丨5mi用之微量離心管中。在 4C、lOOOOrpm之條件下離心處理5分鐘而使細胞不溶物沈 凝，並在測定前將萃取上澄液凍結保存在_8(rc。 19 201138858 浴解已/東結保存之萃取上澄液，並使用檢體稀釋液而 分別加以稀釋至10倍’再藉EGF-R Active EGF receptor EIA 套件之測定法進行測定。 結果則由磷酸化EGF受體標準液之標準曲線算出，並 以EGF受體中之磷酸酪胺酸數(fm〇丨/L)代表之。 結果即如第6圖所示。 培養34小時之檢討中’與未添加控制組(c〇nt)比較，均 發現EGF-R之填酸化作用亢進。燕窩酵素分解物2之〇1 %及 0.01¾添加濃度組（各為Enz20.1%、Εηζ20.01ο/〇)之EGF-R之 填酸化數均為高於作為正控制之r-EGF之〇.15ng/ml添加組 (r-EGF 0.15ng)之值。燕窩酵素分解物1%添加組 (Enz 10.1 %)則在極早期時（丨〇小時）即發現EGF_R之磷酸化 作用完進。 由結果可知，燕高酵素分解物1及燕窩酵素分解物2均 可在0.1%及0.01%之添加濃度下促進EGF-R之磷酸化，而暗 示了對表皮細胞(A431細胞)之增殖確有影響。 C圖式簡單說明】 第1圖係顯示燕窩樣本中之涎酸量之測定結果之圖表。 第2圖係係顯示燕窩樣本中之蛋白質量之測定結果之 圖表。 第3圖係顯示GPC-HPLC之燕窩酵素分解物1之測定結 果之圖表。 第4圖係顯示GPC-HPLC之燕窩酵素分解物2之測定結 果之圖表。 20 201138858 第5圖係顯示第（3)實施例之結果之圖表。 第6圖係顯示第（4)實施例之結果之圖表。 【主要元件符號說明】 (無） 21• Column temperature: 35°C 15 201138858 • Mobile phase: 50mM CH3COONH3 pH6.7 • Flow rate: 〇.4ml/min • s type material injection amount: 1〇μ1 • Detector: UV spectrometer (UV220nm Determination) [Molecular weight marker used] • BSAMW66,000 • Gly-gly MW132.1 - Gly-gly-gly MW 189.17 .L-aspartic acid ( L-Asparaginic acid) MW133.1 • DL-Asparaginic acid MW150.1 • N-Acetylneuraminic acid MW309.3 The results are as follows 3 and 4 The figure shows. Further, the molecular weight of the protein contained in the bird's nest enzyme decomposition product has been evaluated by the Western blot method. Specifically, the amount of protein in the decomposition of the bird's nest enzyme is confirmed by following the above-described measurement method. The protein (1 〇〇μβ) derived from the bird's nest enzyme decomposition product was separated by 10 to 15% of polyacrylamide using SDS-PAGE. The protein dispersed in the colloid was transferred to a lmmunoblot PVDF membrane (p〇lyvinylidene difluoride, available from Bio-Lad Lab). The PVDF membrane was again blocked for 1 hour in PBS containing 3% skim milk and 0.1% Tween-20, and visually judged. Further, when evaluating the molecular weight of a protein, reference has been made to the information disclosed in the well-known literature (Guo, C. T., Suzuki Y., et al. 5 Antiviral Res. 70, pp. 140-146 (2006)). 16 201138858 From the above results, it is known that the amount of protein molecules contained in the enzyme decomposition product of Yan is in the range of about 7G to 12G, and the degree of GGG (otherwise, depending on the evaluation conditions, the upper limit can reach 200,000). Further, from the results obtained, it is known that the average molecular weight of the protein of the bird's nest enzyme decomposition product is about 55, coffee. Further, it is known that the average molecular weight of the protein of the bird's nest enzyme decomposition product 2 is about 33,000. (3) Enzyme-treated cell proliferation-promoting activity of bird's nest E-F F-R has been used to express the skin-constituting cells to review the enzyme-treated EGF effect, that is, cell proliferation-promoting activity. (1) Materials: Cell lines EGF-R derived from human epidermis (skin), which was purchased from ATCC, was used to express cell A431 cells. The bird's nest enzyme decomposition product uses the Yanwan enzyme decomposition product 1 (enzyme treatment for 3 hours), the bird's nest enzyme decomposition product 2 (enzyme treatment for 24 hours), and the bird's nest hot water extract. (ii) Experimental method Add Affluent Enzyme Decomposition or Bird's Nest Hot Water Extract to A431 cells that have been adjusted to 5xl04cells/ml by DME medium supplemented with 3% FCS, and press 5> in a 96-well flat pan. ; <103ceUs / lml / well conditions for culture. Final added concentration: . Bird's nest hot water extract (heart): 1% '〇·1%, 0.01% • Bird's nest hot water extract l (Enzl): 1% ' 0.1%, 0.01% • Bird's nest hot water extract 2 (Enz2): 1%, 0.1%, 0.01% 17 201138858 In addition, a group in which no bird's nest enzyme decomposition product and r_EGF2 were not added were prepared for culture. , : The proliferation of the field cell was long-term use of the Cell Counting Kit-8 (produced by the Tongren Chemical Research Institute Co., Ltd.) (an alternative method for the insertion test of 3H thymidine) to investigate and investigate the presence or absence of EGF-like effects. As a result, ABS (absorbance) represents an amount of pigment which is extremely soluble in the vicinity of 46 〇 nm. The result is as shown in Figure 5. From the results, it was found that the cell proliferation inhibition group and the proliferation group were clearly distinguished from the unadded control group (A431/3% FCS). The proliferation-promoting effect of the addition group of Yanxiang Enzyme Decomposition 1 and Bird's Nest Enzyme Decomposition 2 (especially 0.1% and 0.01%) has been confirmed. However, the 1% addition group of the hot water extract of bird's nest had extremely inhibited cell proliferation even when compared with the untreated control group. In the 1% addition group and the 0.01 addition group of the hot water extract of bird's nest, the cell proliferation was lower than that of the unadded control group. Therefore, it has been confirmed that the decomposition of bird's nest enzymes promotes cell proliferation, while the hot water extract of the bird's nest inhibits or hinders the tendency of cell proliferation. (4) Verification of EGF action such as bird's nest enzyme decomposition product (review of EGF_R_replication) The EGF effect such as bird's nest enzyme decomposition product has been reviewed by using EGF-R to express the phosphorylation of the egf receptor of skin-constituting cells. (1) Materials: Cell lines EGF-R derived from human epidermis (skin), which was purchased from ATCC, was used to express cell A431 cells. The bird's nest enzyme decomposition product is decomposed by the bird's nest enzyme obtained by the above (1). 18 201138858 The substance 1 (enzyme treatment for 3 hours) and the bird's nest enzyme decomposition product 2 (enzyme treatment for 24 hours). Human recombinant EGF (r-EGF) (taken from Protein Express) has been used as a positive control. The acidification of EGF-R was measured using the Active EGF receptor EIA kit (Takara). (ii) Cell culture: A431 cells adjusted to 5 x 1 〇 4 cells/ml were added to the A431 cells which had been adjusted to 5 x 1 〇 4 cells/ml, and the bird's nest enzyme decomposition product was added as a positively controlled r-EGF, and The cells were cultured in a 24-well flat chassis at 104 cells/lml/well. Final concentration: • Bird's Nest Enzyme Decomposition l (Enzl) ·· 0.1%, 〇.〇1〇/0 • • Bird's Nest Enzyme Decomposition 2 (Enz2): 0.1%, 0.01% • r-EGF : 〇.15ng /ml 'An unadded grade of bird's nest enzyme decomposed product and r-EGF was also prepared for culture for comparison. (iv) Adjustment and measurement of the specimen: During the culture period of 10 hours to 50 hours (data is 1 to 34 hours), the cells are recovered for a long period of time, and the receptor extraction buffer attached to the assay kit is used for 6 weeks. The specimen. After removing the culture supernatant, the receptor extraction buffer was added according to 5〇|al/well, and the cell was scraped off using a cell scraper (Ceii scraper), and the cell extract of 2Well fraction was accumulated in the trace amount of 5M. In the centrifuge tube. Centrifuge at 4C, 1000 rpm for 5 minutes to cause cell insolubles to coagulate, and freeze the extract on the extract before storage. _8 (rc. 19 201138858 Bath solution / East knot preservation of the extract liquid And diluted with the sample dilution and diluted to 10 times respectively. The measurement was performed by the EGF-R Active EGF receptor EIA kit. The results were calculated from the standard curve of the phosphorylated EGF receptor standard solution and were evaluated by EGF. The number of phosphotyrosine acid (fm〇丨/L) in the body is represented by the results. The results are shown in Fig. 6. In the 34-hour culture review, EGF- was found in comparison with the unadded control group (c〇nt). The acidification of R is increased. The number of acidification of EGF-R in the 1% and 0.013⁄4 added concentration groups of the bird's nest enzyme decomposition product 2 (Enz20.1%, Εηζ20.01ο/〇) are higher than positive The value of the control group of r-EGF was 15 ng/ml (r-EGF 0.15 ng). The 1% addition group of bird's nest enzyme decomposition product (Enz 10.1%) found EGF_R at very early time (丨〇 hours). The phosphorylation is completed. From the results, it can be seen that Yangao Enzyme Decomposition 1 and Bird's Nest Enzyme Decomposition 2 can be 0.1% and 0.01%. Accelerating the phosphorylation of EGF-R at an increased concentration, suggesting an effect on the proliferation of epidermal cells (A431 cells). Brief Description of C Schematic Figure 1 is a graph showing the results of determination of the amount of citric acid in bird's nest samples. Fig. 2 is a graph showing the results of measurement of the amount of protein in the bird's nest sample. Fig. 3 is a graph showing the measurement results of the bird's nest enzyme decomposition product 1 by GPC-HPLC. Fig. 4 is a bird's nest enzyme showing GPC-HPLC. A graph showing the results of the measurement of the decomposition product 2. 20 201138858 Fig. 5 is a graph showing the results of the (3) example. Fig. 6 is a graph showing the results of the (4) example. No) 21