JP5881186B2 - 生薬抽出物またはその乳酸菌醗酵物を含むアトピー皮膚炎の予防または治療用の組成物 - Google Patents
生薬抽出物またはその乳酸菌醗酵物を含むアトピー皮膚炎の予防または治療用の組成物 Download PDFInfo
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- lactic acid
- acid bacteria
- atopic dermatitis
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Description
前記生薬抽出物は、クジン、カンゾウ、スイカズラ、トウキ、ウド、モッコウ、ボウフウ、サンソウニン、ドクダミ、レンギョウ、ゴボウシ、インヨウカク、オタネニンジン、シコン、チユ、センキュウ、ゲンジンおよびイタドリを抽出して製造される漢方製剤であり、クジン160g、カンゾウ80g、スイカズラ80g、トウキ80g、ウド80g、モッコウ80g、ボウフウ80g、サンソウニン80g、ドクダミ160g、レンギョウ80g、ゴボウシ160g、インヨウカク80g、オタネニンジン160g、シコン80g、チユ80g、センキュウ80g、ゲンジン160gおよびイタドリ80gの割合で生薬混合物を製造した後、前記生薬混合物1840gの10倍量である18.4lの蒸留水に入れて1時間浸漬させた後、115℃で3時間超高速真空低温抽出器(韓国、kyungseo machine社製、cosmos-600)、標準試験ふるい(standard testing sieve、Aperture 500μmと150μm)を用いて煎じ薬を濾過し、本発明の生薬抽出物を製造した。
<2-1>乳酸菌の種菌培養
本発明に係る生薬抽出物の乳酸菌醗酵物の製造に用いられた乳酸菌は次のとおりである。:ラクトバチルスアシドフィルス(KFRI #128)、ラクトバチルスカゼイ(KFRI #127)、ラクトバチルスプランタルム(KFRI #144、KFRI #402)、ラクトバチルスファーメンタム(KFRI #164)、ラクトバチルスブルガリクス(KFRI #344)、ラクトバチルスデルブリュッキー亜種ラクティス(KFRI #442)、ラクトバチルスガッセリ(KFRI #658、KCTC 3163)およびビフィドバクテリウムブレーベ(KFRI #744)。前記菌株は、斜面培地と液体培地で継代して実験に用いた。乳酸菌を斜面培地に接種して37℃の培養器で24時間培養して細菌のコロニーが形成されれば、パラフィンフィルムで酸素を遮断して冷蔵室で保存し、細菌の活性が低下するか雑菌による汚染を防止するために2〜3週に一回ずつ新たな斜面寒天培地に移植した。液体培地に接種して37℃の培養器で24時間培養した。種菌培養にはMRS培地(10g/l Peptone、10g/l Beef extract、5g/l Yeast extract、20g/l Glucose、1ml/l Tween 80、2g/l K2HPO4、5g/l Sodium acetate、2g/l Triammonium citrate、0.2g/l MgSO4 7H2O、0.2g/l MnSO4 4H2O、pH 6.2〜6.6)を用いた。
1M NaOHを用いて実施例1で製造した生薬抽出物のpHを8.0に調整し、121℃、1.5気圧で15分間加圧滅菌して常温まで冷却した。その結果、醗酵前の対照群のpHが8.0から6.3まで下がったが、これは121℃、1.5気圧で15分間滅菌される高温高圧条件で生薬抽出物に存在する食物繊維成分の一部が有機酸に分解されてpHが低下したためであると推定される。生薬抽出物に1%ボリューム(v/v)の前記乳酸菌(1-5×108CFU/ml)を添加し、37℃で48時間通気培養して液体醗酵させた後、前記標準試験ふるいで濾過して生薬抽出物の乳酸菌醗酵物を製造した。前記乳酸菌醗酵物のpHおよび乾物量変化は下記表1のとおりである。
前記で製造された本発明に係る生薬抽出物の乳酸菌液体醗酵物を遠心分離機を用いて残渣を除去し、0.45μmの濾過器を用いて濾過した後、下記表2の条件に従ってHPLC分析を行った。
本発明の実験例では、韓国の食品医薬品安全庁のバイオ生薬局で提示した『生薬漢方薬製剤の効力試験ガイドライン-アトピー皮膚炎』(ガイドライン管理番号1-019-2010-0000(0))の指針に従い下記実験を行った。実験動物を飼育ケースにそれぞれ入れて、23±3℃の温度と50±5%の湿度で12時間の間隔で明-暗サイクルを与えながら飼育した。基本的な実験食は、American Institute Nutrition(1970)で勧めるものに基づき製造して用いた。試験物質を液体溶媒(飲水)に混合して実験を行い、韓国の食品医薬品安全評価機関の標準作業手順書の『試験物質の配合および調剤(NITR/SOP/TSB/002_02)』に従って実験を行った。また、各容量群が均質性のある個体群で構成されるように食品医薬品安全評価機関の標準作業手順書の『げっ歯類の群、飼育ケースおよび飼育ケース棚の無作為割付(NITR/SOP/GTX/002_02)』に従って実験群を分離した。試験結果の統計的有意性を評価するために、各試験群当たりの個体数は5〜7匹にし、試験薬物と対照群の変化数値および誤差をウイルコクスン順位和検定法(マン-ホイットニー検定)を用いて有意性を検証した。統計プログラムとしてはSASを用いた。
本評価方法は、アトピー皮膚炎で通常用いられる臨床的目視評価法であり、アトピー皮膚炎の深刻度を以下の5項目それぞれを評価した点数の総合で示した。評価項目は、紅斑、掻痒症と皮膚乾燥、浮腫と血腫、糜爛および苔癬化である。それぞれの項目に対して症状がない(0点)、症状が軽い(1点)、普通(2点)、症状がひどい(3点)と採点した後、5項目の点数を合算することで最小0点(何れの症状もない状態)から最高15点(全ての項目の症状がひどい状態)までの評価点数を付与した。
環境的な刺激によってアトピー皮膚炎が発生したマウスは、対照群に比べて週齢の経過とともに血中IgE濃度が急激に増加すると予想した。環境的なストレスによるTh1/Th2免疫反応の変化を確認するために、マウスから時間ごとに血液を採取して血漿を分離し、ELISAキットを用いてIgE濃度を測定することで試験物質が血中IgE濃度が減少または抑制したか否かを評価した。
<1-1>シロップ剤の製造
生薬抽出物または生薬抽出物の醗酵物を有効成分2%(重量/体積)として含有するシロップを次のような方法で製造した。まず、実施例1または実施例2で製造した生薬抽出物または生薬抽出物の醗酵物粉末、サッカリン、糖を温水80gに溶解した。前記溶液を冷却させた後、これにグリセリン、サッカリン、香味料、エタノール、ソルビン酸および蒸留水からなる溶液を製造して混合した。この混合物に水を添加して100mlになるようにした。
生薬抽出物または生薬抽出物の醗酵物 2g
サッカリン 0.8g
糖 25.4g
グリセリン 8.0g
香味料 0.04g
エタノール 4.0g
ソルビン酸 0.4g
蒸留水 定量
有効成分15mgが含有された錠剤を次のような方法で製造した。
実施例1または実施例2で製造した生薬抽出物または生薬抽出物の醗酵物250gをラクトース175.9g、ジャガイモデンプン180gおよびコロイド状ケイ酸32gと混合した。前記混合物に10%ゼラチン溶液を添加した後、粉砕して14メッシュのふるいを通過させた。これを乾燥させ、これにジャガイモデンプン160g、タルク50gおよびステアリン酸マグネシウム5gを添加して得た混合物を錠剤に製造した。
生薬抽出物または生薬抽出物の醗酵物 250g
ラクトース 175.9g
ジャガイモデンプン 180g
コロイド状ケイ酸 32g
10%ゼラチン溶液ジャガイモデンプン 160g
タルク 50g
ステアリン酸マグネシウム 5g
<2-1>食品の製造
実施例1または実施例2で製造した生薬抽出物または生薬抽出物の醗酵物を含む食品を次のように製造した。
生薬抽出物または生薬抽出物の醗酵物20〜95重量%で健康増進用の料理用調味料を製造した。
生薬抽出物または生薬抽出物の醗酵物0.2〜1.0重量%をトマトケチャップまたはソースに添加して健康増進用のトマトケチャップまたはソースを製造した。
生薬抽出物または生薬抽出物の醗酵物0.5〜5.0重量%を小麦粉に添加し、この混合物を用いてパン、ケーキ、クッキー、クラッカーおよび麺類を製造して健康増進用の小麦粉食品を製造した。
生薬抽出物または生薬抽出物の醗酵物0.1〜5.0重量%をスープおよび肉汁に添加して健康増進用の食肉加工製品、麺類のスープおよび肉汁を製造した。
生薬抽出物または生薬抽出物の醗酵物10重量%を牛挽肉に添加して健康増進用の牛挽肉を製造した。
生薬抽出物または生薬抽出物の醗酵物5〜10重量%を牛乳に添加し、前記牛乳を用いてバターおよびアイスクリームのような様々な乳製品を製造した。
玄米、麦、もち米、ハトムギを公知の方法でアルファ化して乾燥したものを焙煎した後、粉砕機で粒度60メッシュの粉末に製造した。黒豆、黒ゴマ、エゴマも公知の方法で蒸して乾燥したものを焙煎した後、粉砕機で粒度60メッシュの粉末に製造した。生薬抽出物または生薬抽出物の醗酵物を真空濃縮機で減圧濃縮し、噴霧乾燥器、熱風乾燥器で乾燥して得た乾燥物を粉砕機で粒度60メッシュに粉砕して乾燥粉末を得た。前記で製造した穀物類、種実類および生薬抽出物または生薬抽出物の醗酵物の乾燥粉末を以下の割合で配合して製造した。
種実類(エゴマ7重量%、黒豆8重量%、黒ゴマ7重量%)、
生薬抽出物または生薬抽出物の醗酵物の乾燥粉末(3重量%)、
レイシ(0.5重量%)、
ジオウ(0.5重量%)
1.炭酸飲料の製造
砂糖 5〜10%、クエン酸 0.05〜0.3%、キャラメル 0.005〜0.02%、ビタミンC 0.1〜1%の添加物を混合し、これに79〜94%の精製水を混合してシロップを製造し、前記シロップを85〜98℃で20〜180秒間殺菌して冷却水と1:4の割合で混合した後、炭酸ガスを0.5〜0.82%注入して生薬抽出物または生薬抽出物の醗酵物を含有する炭酸飲料を製造した。
異性化糖(0.5%)、オリゴ糖(2%)、砂糖(2%)、食塩(0.5%)、水(75%)のような副材料と生薬抽出物または生薬抽出物の醗酵物を均質に配合して瞬間殺菌をした後、これをガラスボトル、ペットボトルなどの小容器で包装して健康飲料を製造した。
生薬抽出物または生薬抽出物の醗酵物5gをトマトジュースまたはニンジンジュース1000mlに加えて健康増進用の野菜ジュースを製造した。
生薬抽出物または生薬抽出物の醗酵物1gをリンゴジュースまたはブドウジュース1000mlに加えて健康増進用の果汁を製造した。
<3-1>乳化剤形の化粧品の製造
表3に記載の組成で乳化剤形の化粧品を製造した。製造方法は下記のとおりである。
1)1〜9の原料を混合した混合物を65〜70℃に加熱した。
2)10の原料を前記段階1)の混合物に投入した。
3)11〜13の原料の混合物を65〜70℃に加熱して完全に溶解させた。
4)前記段階3)を実行しながら前記2)の混合物を徐々に添加して8000rpmで2〜3分間乳化させた。
5)14の原料を少量の水に溶解させた後、前記段階4)の混合物に添加して2分間さらに乳化させた。
6)15〜17の原料をそれぞれ坪量した後、前記段階5)の混合物に入れて30秒間さらに乳化させた。
7)前記段階6)の混合物を乳化させた後、脱気過程を経て25〜35℃に冷却させることで乳化剤形の化粧品を製造した。
Claims (8)
- クジン、カンゾウ、スイカズラ、トウキ、ウド、モッコウ、ボウフウ、サンソウニン、ドクダミ、レンギョウ、ゴボウシ、インヨウカク、オタネニンジン、シコン、チユ、センキュウ、ゲンジンおよびイタドリのみからなる生薬混合物に水または有機溶媒を加えた後、抽出して製造される生薬抽出物に乳酸菌を接種して醗酵させることにより製造されることを特徴とする、生薬抽出物のアトピー皮膚炎の予防または治療用の乳酸菌醗酵物の製造方法。
- 前記乳酸菌は、ラクトバチルス属、ビフィドバクテリウム属、ストレプトコッカス属、ロイコノストック属、ペディオコッカス属およびラクトコッカス属の微生物からなる群から選択されることを特徴とする、請求項1に記載の乳酸菌醗酵物の製造方法。
- 前記乳酸菌は、ラクトバチルスアシドフィルス種の微生物であることを特徴とする、請求項2に記載の乳酸菌醗酵物の製造方法。
- 前記乳酸菌は、試料重量に対して0.5〜5重量%の量で接種することを特徴とする、請求項1〜3の何れか一項に記載の乳酸菌醗酵物の製造方法。
- 前記乳酸菌は、20〜40℃の温度で24〜52時間醗酵させることを特徴とする、請求項1〜3の何れか一項に記載の乳酸菌醗酵物の製造方法。
- 請求項1に記載の製造方法で乳酸菌醗酵物を製造する工程と、
前記乳酸菌醗酵物を添加する工程と、を含む、
アトピー皮膚炎の予防または治療用の薬学的組成物の製造方法。 - 請求項1に記載の製造方法で乳酸菌醗酵物を製造する工程と、
前記乳酸菌醗酵物を添加する工程と、を含む、
アトピー皮膚炎の予防または改善用の健康食品の製造方法。 - 請求項1に記載の製造方法で乳酸菌醗酵物を製造する工程と、
前記乳酸菌醗酵物を添加する工程と、を含む、
アトピー皮膚炎の予防または改善用の化粧料組成物の製造方法。
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KR20100093151A (ko) | 2009-02-16 | 2010-08-25 | 장문식 | 아토피성 피부염의 예방 효과를 가지는 조성물 |
KR20090125726A (ko) | 2009-10-22 | 2009-12-07 | 신화제약 (주) | 호장근 발효 추출물 및 이를 함유하는 피부노화방지 한방 화장료 조성물 |
CN101804166B (zh) * | 2010-05-05 | 2011-07-27 | 余静 | 一种治疗小儿皮炎湿疹的中药组合物 |
CN101810799B (zh) * | 2010-05-26 | 2011-06-01 | 泰一和浦(北京)中医药研究院有限公司 | 一种治疗出疹的中药组合物及其制备方法 |
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- 2010-10-29 US US13/825,839 patent/US9295704B2/en active Active
- 2010-10-29 EP EP10857921.0A patent/EP2623108B1/en active Active
- 2010-10-29 CN CN201080069308.1A patent/CN103228287B/zh active Active
- 2010-10-29 JP JP2013530081A patent/JP5881186B2/ja active Active
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WO2012043920A1 (ko) | 2012-04-05 |
EP2623108A4 (en) | 2014-03-12 |
KR101226758B1 (ko) | 2013-01-25 |
EP2623108A1 (en) | 2013-08-07 |
KR20120032311A (ko) | 2012-04-05 |
US20130224181A1 (en) | 2013-08-29 |
JP2013538824A (ja) | 2013-10-17 |
EP2623108B1 (en) | 2017-09-27 |
US9295704B2 (en) | 2016-03-29 |
CN103228287B (zh) | 2015-12-02 |
CN103228287A (zh) | 2013-07-31 |
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