JP2018070452A - 前駆脂肪細胞分化促進剤 - Google Patents
前駆脂肪細胞分化促進剤 Download PDFInfo
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Abstract
【解決手段】豆乳醗酵液、クロレラ抽出物に優れた前駆脂肪細胞分化促進作用を有することを見い出し、前駆脂肪細胞分化促進剤、更に前駆脂肪細胞分化促進剤を含有してなる各種組成物(皮膚化粧料、頭髪化粧料、飲食品組成物、医薬品組成物)。
【効果】各種組成物は、加齢に伴って生じるしわ、たるみ等の皮膚状態を予防又は改善、肌の張りの改善やバストアップ、毛包の再生に伴って生じる脱毛の防止、育毛作用、養毛作用を促す等の頭髪状態の改善、又は、肥満や高血圧、高脂血症、動脈硬化症、糖尿病等の生活習慣病等の予防又は改善に役立つものである。
【選択図】なし
Description
ン脂質、リゾレシチン、水素添加大豆リン脂質、部分水素添加大豆リン脂質、水素添加卵黄リン脂質、部分水素添加卵黄リン脂質、水酸化レシチン等のリン脂質類;シリコーン系両性界面活性剤等;高分子界面活性剤では、ポリビニルアルコール、アルギン酸ナトリウム、デンプン誘導体、トラガントガム、アクリル酸・メタアクリル酸アルキル共重合体、2−メタクロイルオキシエチルホスホリルコリン・ステアリルメタクリレート共重合体;シリコーン系各種界面活性剤が好ましいものとして挙げられる。
大豆(黄大豆)を原料として得た豆乳(200ml)に、乳酸菌(Lactobacillus bulgaricus)を滅菌水又は精製水に溶解もしくは懸濁させて接種する。次に、室温約40℃、約15時間醗酵させた後、放冷し、殺菌後、濾過して約160mlの豆乳醗酵液を得る。
大豆(黒大豆)を原料として得た豆乳(200ml)に、乳酸菌(Lactobacillus bulgaricus)を滅菌水又は精製水に溶解もしくは懸濁させて接種する。次に、室温約37℃、約20時間醗酵させた後、放冷し、殺菌後、濾過して約160mlの豆乳醗酵液を得る。
大豆(赤大豆)を原料として得た豆乳(200ml)に、乳酸菌(Lactobacillus bulgaricus)を滅菌水又は精製水に溶解もしくは懸濁させて接種する。次に、室温約38℃、約10時間醗酵させた後、放冷し、殺菌後、濾過して約150mlの豆乳醗酵液を得る。
大豆(青大豆)を原料として得た豆乳(200ml)に、乳酸菌(Lactobacillus bulgaricus)を滅菌水又は精製水に溶解もしくは懸濁させて接種する。次に、室温約40℃、約5時間醗酵させた後、放冷し、殺菌後、濾過して約150mlの豆乳醗酵液を得る。
クロレラ・ブルガリスの全草1kgに12kgの精製水を加えて約3時間80℃で加熱抽出し、濾過した後、得られた濾液に1,3-ブチレングリコールを添加し、約0〜5℃にて10日間静置した後、再度濾過して、約10kgのクロレラ抽出液を得る。
クロレラ・ピレノイドサの全草1kgに12kgの精製水を加えて約3時間80℃で加熱抽出し、濾過した後、得られた濾液に1,3-ブチレングリコールを添加し、約0〜5℃にて10日間静置した後、再度濾過して、約10kgのクロレラ抽出液を得る。
クロレラ・エリプソイデイアの全草0.5kgに6kgの精製水を加えて約4時間80℃で加熱抽出し、濾過した後、得られた濾液に70%グリセリン水溶液を添加し、約0〜5℃にて10日間静置した後、再度濾過して、約6kgのクロレラ抽出液を得る。
マウス由来前駆脂肪細胞3T3-L1細胞は、dexamethasone(DEXA)、3-isobutyl-1-methylxanthine(IBMX)、insulinによる刺激で効率良く脂肪細胞に分化するため、研究に広く用いられている。3T3-L1細胞は分化すると、紡錘状の形態から丸みを帯びた形へと変化し、細胞内に微細少量の脂肪滴が生じる。分化が進むと、脂肪滴は大きくなり、Oil Red Oで染色すると赤色に染まる。イソプロパノールを加えて色素を抽出し、その吸光度を測定し、脂肪蓄積量として分化の程度を評価した。
マウス由来の前駆脂肪細胞、3T3−L1細胞を使用した。尚、培養液には、10%CS(ATCC 30-2030)を含むD−MEM培地を使用し、5%CO2、37℃の条件で培養した。
前記細胞を24ウェルプレート(コーニング)に1ウェル当たりの細胞数が1×104個となるように播種し、コンフルエントに達したらD−MEM/10%FBS培地に交換した。
コンフルエント2日後に分化誘導培地(0.25mM Dexamethasone, 0.5mM 3-isobutyl-1-methylxanthine, 5mg/ml Insulinを含むD−MEM/10%FBS培地)に交換すると同時に、試料を添加した。この日を分化誘導0日目とする。その2日後(分化誘導2日目)に5mg/ml Insulin含有培地に交換した。さらにその2日後(分化誘導4日目)に維持培地に交換し、分化誘導8日目まで培養した。分化誘導0日目から8日目までの期間、試料を添加した。
Oil Red O染色:分化誘導8日目の細胞をリン酸緩衝液(pH7.4)で洗浄後、10%中性ホルマリンで固定した。蒸留水で洗浄後、ろ過した0.3% Oil Red Oイソプロパノール液で染色した。蒸留水で洗浄後、イソプロパノールで色素を抽出した。510nmでの吸光度を測定し、対照を脂肪蓄積量100%として換算して評価を行った。結果を図1に示した。
「試料」
本試験の試料は、製造例2で得られた豆乳醗酵液を1.0%、又は、2.0%添加、製造例5で得られたクロレラ抽出液を1.0%添加して試験に供した。尚、豆乳醗酵液の対照として精製水を添加し、クロレラ抽出液の対照として1,3-ブチレングリコール溶液を、終濃度が0.03%となるよう添加した。
図1に示すように、対照の脂肪蓄積量が100%に対して、本発明の豆乳醗酵液では、1.0%添加時で108%、2.0%添加時で115%の脂肪蓄積量の増加が認められ、クロレラ抽出液では、1.0%添加時で129%の脂肪蓄積量の増加が認められた。よって、前駆脂肪細胞の成熟脂肪細胞への分化促進作用を有していることが確認でき、前駆脂肪細胞分化促進剤として利用できる。
(1)皮膚一次刺激性試験
製造例1〜7によって得られた本発明の豆乳醗酵液又はクロレラ抽出液を乾燥固形分濃度が約1.0%となるように精製水にて調整し、背部を剪毛した日本白色家兎(雌性,1群3匹,体重2.3kg前後)の皮膚に適用した。判定は、適用後24、48、72時間に一次刺激性の評点法にて紅斑及び浮腫を指標として行った。その結果は、すべての動物において、何など、紅斑及び浮腫を認めず陰性と判定され、安全であると判断された。
(2)急性毒性試験
同様に製造例1〜7によって得られた本発明の豆乳醗酵液又はクロレラ抽出液(固型分濃度が約5w/w%になるように精製水にて調整)を減圧濃縮・乾燥して得られた粉末を試験前、4時間絶食させたddy系マウス(雄性及び雌性,1群5匹,5週齢)に2,000mg/kg量経口投与し、毒性症状の発現、程度などを経時的に観察した。その結果、すべてのマウスにおいて14日間何等異状を認めず、又、解剖の結果も異状がなかった。よって、LD50は2,000mg/kg以上であり、安全であると判断された。
上記の評価結果に従い、以下にその処方例を示すが、各処方例は各製品の製造における常法により製造したもので良く、配合量のみを示した。又、本発明はこれらに限定されるわけではない。
同様の結果が得られた。
ガーゼ又はリニメント布に豆乳醗酵液、クロレラ抽出液、抗生物質、抗炎症剤等、適量を混合した処方液を含浸させ、外傷部に添付する。又、豆乳醗酵液、又は、クロレラ抽出液を直接、局所に散布し、ガーゼ等で被覆しても良い。
本発明の製造例2の豆乳醗酵液 3.0
クエン酸 1.0
果糖ブドウ糖液糖 70.0
香料 適量
精製水 100とする残余
本発明の製造例5のクロレラ抽出液 3.0
クエン酸 1.0
果糖ブドウ糖液糖 70.0
香料 適量
精製水 100とする残余
スクワラン 5.0
オリーブ油 5.0
ホホバ油 5.0
セチルアルコール 1.5
グリセリンモノステアレート 2.0
ポリオキシエチレン(20)セチルエーテル 3.0
ポリオキシエチレン(20)ソオルビタンモノオレート 2.0
1,3-ブチレングリコール 1.0
グリセリン 2.0
本発明の製造例3の豆乳醗酵液 5.0
サクラの葉50%エタノール抽出液 3.0
防腐剤(パラオキシ安息香酸エステル) 適量
香料(バラ水) 適量
精製水 100とする残余
グリセリン 5.0
1,3-ブチレングリコール 5.0
モノラウリン酸ホ゜リオキシエチレンソルヒ゛タン(20E.O) 1.0
エタノール 15.0
本発明の製造例5のクロレラ抽出液 5.0
マンダリンオレンジ果皮30%エタノール抽出液 5.0
ボダイジュ花又は果実50%1,3-ブチレングリコール抽出液 5.0
アミノ酸(アルギニン、グリシン、グルタミン等) 0.5
抗菌(ラクトフェリン溶液) 適量
防腐剤(エチルパラベン) 適量
香料(西洋薄荷水) 適量
精製水 100とする残余
ポリビニルアルコール 15.0
キサンタンガム 0.4
ヒドロキシエチルセルロース 0.2
1,3-ブチレングリコール 5.0
本発明の製造例1の豆乳醗酵液 5.0
本発明の製造例6のクロレラ抽出液 5.0
葛根50%プロピレングリコール抽出液 0.5
グリチルリチン酸ジカリウム 0.2
防腐剤(サルチル酸) 適量
香料(葡萄水) 適量
精製水 100とする残余
エタノール 40.0
オレイン酸エチル 1.0
ポリオキシエチレン(40)硬化ヒマシ油 2.0
本発明の製造例4の豆乳醗酵液 5.0
オウゴン根茎又は根皮50%エタノール抽出液 1.0
枸杞根茎又は根皮50%エタノール抽出液 1.0
防腐剤(パラベン) 適量
香料(セージ水) 適量
精製水 100とする残余
エタノール 60.0
本発明の製造例6のクロレラ抽出液 5.0
タチジャコウソウ全草50%プロピレングリコール抽出液 2.0
ビタミンE誘導体 0.5
トウガラシチンキ 0.5
レゾルシン 0.5
グリチルリチン酸ジカリウム 0.5
カルボキシメチルキチン溶液 0.5
コラーゲン蛋白質酵素分解液 2.0
アミノ酸(アルギニン、グリシン、グルタミン等) 0.5
抗菌・防腐剤(プロピルパラベン) 0.1
香料(ローズマリー水) 適量
精製水 100とする残余
エタノール 40.0
オレイン酸エチル 1.0
ポリオキシエチレン(40)硬化ヒマシ油 2.0
本発明の製造例3の豆乳醗酵液 2.5
本発明の製造例7のクロレラ抽出液 2.5
コンフリー60%エタノール抽出液 1.0
サボンソウ葉50%1,3-ブチレングリコール抽出液 1.0
グリチルリチン酸 0.5
防腐剤(エチルパラベン) 適量
香料(緑茶エキス) 適量
精製水 100とする残余
ラウリル硫酸トリエタノールアミン 5.0
ポリオキシエチレンラウリルエーテル硫酸ナトリウム 12.0
1,3-ブチレングリコール 4.0
ラウリン酸ジエタノールアミド 2.0
エデト酸二ナトリウム 0.1
本発明の製造例4の豆乳醗酵液 4.0
ハマメリス樹皮熱水抽出液 2.0
当帰根30%1,4-ブチレングリコール抽出液 1.0
コーヒー豆50%1,3-ブチレングリコール抽出液 2.0
ヒドロキシプロピルキトサン溶液 1.0
ムコ多糖体溶液 1.0
グリチルリチン酸ジカリウム 0.1
ジンクピリチオン 0.1
抗菌・防腐剤(感光素101号) 適量
pH調整剤 適量
香料(白檀水) 適量
精製水 100とする残余
塩化ステアリルトリメチルアンモニウム 2.0
セトステアリルアルコール 2.0
ポリオキシエチレンラノリンエーテル 3.0
プロピレングリコール 5.0
エチレングリコール 3.0
本発明の製造例5のクロレラ抽出液 4.0
コボタンヅル全草50%1,2-ペンタンジオール抽出液 2.0
西洋菩提樹花50%1,4-ブチレングリコール抽出液 2.0
オリーブ葉50%1,2-ブチレングリコール抽出液 2.0
甘茶全草50%グリセリン抽出液 2.0
ビフィズス菌醗酵液 2.0
アルギン酸ナトリウム 2.0
トレハロ−ス溶液 1.0
加水分解ケラチン溶液 1.0
抗菌・防腐剤(塩化ベンザルコニウム) 適量
香料(ハマメリス水) 適量
精製水 100とする残余
本発明の製造例5のクロレラ抽出液乾燥粉末 1.0
マンダリンオレンジ果実熱水抽出液乾燥粉末 1.0
乳糖 30.0
コーンスターチ 57.0
結晶セルロース 7.0
ポリビニールピロリドン 4.0
香料 適量
組成:本発明の製造例2の豆乳醗酵液の濃縮物30mg、ヒドロキシプロピルセルロース20mg、軽質無水ケイ酸3mg、乳糖 8mg、結晶セルロース9mg、タルク10mg、ジアシルグリセロール20mgを定法に従って1錠100mgの錠剤を製造した。
本発明の製造例2の豆乳醗酵液乾燥粉末 3.0
セルロース 97.0
香料 適量
本発明の製造例4の豆乳醗酵液乾燥粉末 10.0
本発明の製造例5のクロレラ抽出液乾燥粉末 1.0
ヘスペリジン 80.0
澱粉 9.0
香料 適量
小麦粉 75.0
ベーコン 10.0
鱈子 13.0
本発明の製造例6のクロレラ抽出液乾燥粉末 1.0
食塩・コショウ 適量
バター 適量
青海苔 適量
オリ−ブ油 適量
ブドウ糖液糖 30.0
グレープフルーツ果汁 55.0
ルテオリン-7-O-グルコシド 5.0
本発明の製造例3の豆乳醗酵液乾燥粉末 1.0
ムコ多糖体溶液 3.0
オレンジ果汁 2.0
香料 適量
酸味料 適量
Claims (5)
- 豆乳醗酵液、クロレラ抽出物を有効成分として1種以上を含有する前駆脂肪細胞分化促進剤。
- 請求項第1項記載の前駆脂肪細胞分化促進剤を含有することを特徴とする皮膚化粧料。
- 請求項第1項記載の前駆脂肪細胞分化促進剤を含有することを特徴とする頭髪化粧料。
- 請求項第1項記載の前駆脂肪細胞分化促進剤を含有することを特徴とする飲食品組成物。
- 請求項第1項記載の前駆脂肪細胞分化促進剤を含有することを特徴とする医薬品組成物。
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JP7495172B1 (ja) | 2023-12-12 | 2024-06-04 | 株式会社東洋新薬 | 経口組成物 |
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