JP3852262B2 - Ceramide synthesis accelerator, skin cosmetic and bath agent - Google Patents
Ceramide synthesis accelerator, skin cosmetic and bath agent Download PDFInfo
- Publication number
- JP3852262B2 JP3852262B2 JP2000071647A JP2000071647A JP3852262B2 JP 3852262 B2 JP3852262 B2 JP 3852262B2 JP 2000071647 A JP2000071647 A JP 2000071647A JP 2000071647 A JP2000071647 A JP 2000071647A JP 3852262 B2 JP3852262 B2 JP 3852262B2
- Authority
- JP
- Japan
- Prior art keywords
- skin
- simon
- extract
- ceramide
- ceramide synthesis
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Lifetime
Links
Landscapes
- Cosmetics (AREA)
- Medicines Containing Plant Substances (AREA)
Description
【0001】
【発明の属する技術分野】
本発明は、皮膚表皮層内部で表皮細胞自身のセラミド合成を活発化させ、皮膚バリアー機能を改善することにより、荒れ肌改善及び各種皮膚疾患の改善が期待されるセラミド合成促進剤に関する。また、皮膚化粧料及び入浴剤に関する。
【0002】
【従来の技術】
脂質の一種であるセラミドは、生体内で大部分を占めるグリセロ脂質に比べて量的には少ないが、重要な生理的役割を持つことが最近知られてきている。ヒトを始めとする哺乳類の生体内分布においても生理的に要となっている場所にあるが、中でも脳、肝臓、皮膚等に蓄積されていることが知られている。
【0003】
皮膚では特に表皮角質層にセラミドが集積しているが、これは表皮細胞によって合成分泌され、細胞間に独特のラメラ構造を形成している細胞間脂質の主要成分となっている(Lukas Landman :Anat.Embryol.178,1−3,1988)。
【0004】
角質層は皮膚の保湿能や生体の物理的保護を始めとする一連の生理的役割、いわゆるバリアー機能を持っているが、細胞間脂質はこのバリアー機能の実体であり、生命維持において最も重要な役割の一つを担っている(芋川玄爾:香粧会誌、1(4)、250−253,1991)。この意味から、皮膚セラミドは生体防御の要の1つになっていると言える。
【0005】
肌荒れや乾燥肌、また各種皮膚疾患では、この角質層の健全な形成が妨げられ,バリアー機能低下の起きていることが数多く報告されている。具体的な例としては、皮膚表面の加齢に伴う表皮層のターンオーバーの低下、又は光や温度、気象条件等の外的要因によって生じる肌荒れや乾燥肌が挙げられる。これはバリアー機能の低下が生じ、本来皮膚が有している保水能力の低下と水分蒸散量の増加が生じた結果、誘発されると考えられている(赤崎秀一ほか:日皮会誌、98(1)、41−45,1988)。
【0006】
また、皮膚疾患の中で、アトピー性皮膚炎では患者の炎症部のみならず非炎症部でもバリアー機能の低下や崩壊が見られ、患者の皮膚中、セラミドの全般的な、又は特定の種類の含有量低下が報告されている(川島真:香粧会誌、15(4)、261−262,1991)。
【0007】
この他、乾癬でも患者皮膚中のセラミド量の変動が報告されており(Strfania Motta他:Arch.Dermatol.130,452−456、1994)、この場合もこの変動がバリアー崩壊と関係していると考えられる。
【0008】
このような皮膚バリアーの低下や崩壊からくる皮膚の疾患や不全に対しては、従来、保湿成分の投与で皮膚の乾燥状態を防ぎ潤いを持たせることや、抗炎症剤による湿疹の抑制が試みられてきた。しかし、これらの方法は、角質層表面の水分又は保湿成分の一部を補給する為にその効果が一時的なものに留まり、皮膚内部に充分な潤いを持続的に与えることができなかったり(武村俊之:ファルマシア、28、1、1992)、一時的な炎症を抑えても効果の持続性や副作用に問題のあることが多かった。
【0009】
これに対し、最近バリアー構成主要成分であるセラミドの外部補給で皮膚の改善治療を試みることも行われ、肌荒れ状態やアトピー性皮膚炎に有効なことも報告された(檜垣祐子他:アレルギーの臨床、13(12)、26−28,1993)。
【0010】
しかしながら、この方法は効果の発現が早いと思われる半面、従来用いられていた保湿剤等と同様、効果の持続性の点で不充分であり、皮膚の状態によっては経皮吸収の低下で効果発現が不充分になる等の欠点も考えられる。
【0011】
一方、シモン抽出物の表皮細胞に関する作用は特に知られていなかった。
【0012】
【発明が解決しようとする課題】
本発明者等は係る事情に鑑み、皮膚表皮層内部で表皮細胞自身のセラミド合成を活発化させ、皮膚バリアー機能を改善させることを意図し、鋭意検討した結果、シモン抽出物が有効なセラミド合成促進作用を有することを見出し、本発明を完成するに至ったものであって、その目的とするところは、皮膚表皮層内部で表皮細胞自身のセラミド合成を活発化することにより荒れ肌の改善及び各種皮膚疾患の治療が期待されるセラミド合成促進剤、皮膚化粧料及び入浴剤を提供することにある。
【0013】
【課題を解決するための手段】
上述の目的は、シモン抽出物からなるセラミド合成促進剤、及びシモン抽出物を含有する皮膚化粧料及び入浴剤によって達成される。
【0014】
【発明の実施の形態】
本発明に用いられるシモンは、主として中南米産で栽培されているヒルガオ科サツマイモ属に属する白さつま芋の一種である。極めて繁殖しやすく、容易に入手することができる。従来よりビタミンKやビタミン類、ミネラル類が豊富であることが知られており、ビタミンK・D含有加工食品として販売されている。
【0015】
本発明に用いられるシモン抽出物としては、シモンの葉、茎や根塊(芋)より、各種の溶媒で抽出して得ることができる。また、市販の工業用抽出エキスを用いることもでき、例えば、工業用シモン葉抽出エキス(熊本製粉社製)、工業用シモン芋抽出エキス(熊本製粉社製)等が挙げられる。
【0016】
シモン抽出物を得るための溶媒としては、水、メタノール、エタノール、イソプロピルアルコール等のアルコール類、エチレングリコール、プロピレングリコール、1,3−ブチレングリコール等の多価アルコール類、アセトン等のケトン類、酢酸エチル等のエステル類、ジエチルエーテル等のエーテル類、及びベンゼン等の芳香族化合物を用いることができ、これらの中より二種以上を組み合せて用いることもできる。
【0017】
また、抽出時間は、常温抽出、又は加熱抽出が用いられ、抽出時間に制限はないが、一般的に1時間から1週間が好ましい。
【0018】
シモンは一般的には全草もしくは葉や茎等を個別に乾燥、もしくは生植物をそのまま、又は裁断して抽出に供する。そして、一般的には、シモンの乾物換算5〜50部に対し上記抽出溶媒100部が用いられる。
【0019】
シモン抽出物は、抽出液を濾過しその濾液をそのまま使用してもよいが、通常、濾液を常圧、もしくは減圧下濃縮した濃縮液か、又は更に溶媒を蒸発乾固させて固形物を使用するのが一般的である。
【0020】
本発明に係るセラミド合成促進剤は、化粧料、医薬部外品、医薬品等人体に適用するものの他、飲料、食品等摂取するものに何ら制限なく配合できる。
【0021】
シモン抽出物の皮膚化粧料への配合量は、皮膚化粧料総量を基準としてシモン葉エタノール抽出物の場合は、乾固物換算で、全組成の0.001〜5質量%(以下、%で示す)が好ましく、特に0.01〜0.3%が好ましい。セラミド合成をより効果的に発揮させるためには0.001%以上が好ましく、配合する組成物の色や匂いと調和するという点では5%以下が好ましい。これらのシモン抽出物を含有する化粧料又は入浴剤には、通常医薬品や医薬部外品、皮膚化粧料や入浴剤に用いられる他の成分を同時に配合することができる。
【0022】
本発明に係る皮膚化粧料及び入浴剤の形態は特に限定されるものではないが、例えば、水溶液、アルコール水溶液、エマルジョン、軟膏、、錠剤、パウダー、乳液、又はローション等の一般的な皮膚化粧料又は入浴剤の形態とすることができる。
【0023】
【実施例】
以下、実施例により本発明を更に詳細に説明する。尚、実施例に先立ち、セラミド合成促進剤の評価方法を述べる。
【0024】
セラミド合成促進試験方法
(a)培養表皮細胞
ヒト正常表皮角化細胞は市販品(Epidercell/新生児***:クラボウ社製)を用いた。
【0025】
(b)細胞培養用培地
培地としては MCDB153HAA(和光純薬社より購入)をベースとし、ハイドロコーチゾン(0.5μmol/L)、エタノールアミン(0.1mmol/L)、ホスホエタノールアミン(0.1mmol/L)、インシュリン(5μg/mL)、及びEGF(上皮細胞成長因子:10ng/mL)を加えたK−GM培地を用いた。また、細胞の増殖培養時には、これにBPE(牛脳下垂体抽出液、クラボウ社より購入)を培地1mL当たり4μL添加した。
【0026】
(c)Hepes緩衝液の調製
Hepes(株式会社同仁化学研究所製)7.15g、グルコース(関東化学社製)1.8g、塩化カリウム(関東化学社製)0.22g、塩化ナトリウム(関東化学社製)7.7g、リン酸水素二ナトリウム・12水和物(関東化学社製)0.27gを精製水に溶解し、1mol/L水酸化ナトリウム水溶液にてpH7.4に調整後1Lにメスアップした。
【0027】
(d)トリプシン溶液
0.025%トリプシン(シグマ社製)及び0.01%エチレンジアミン四酢酸二ナトリウム(関東化学社製)を含有するHepes緩衝液を調製した。
【0028】
(e)トリプシン反応停止溶液
(c)で調製したHepes緩衝液に大豆トリプシンインヒビター(以下SBTIと示す、シグマ社製)を溶解し(0.2%)、SBTI溶液としてトリプシン反応の停止に用いた。
【0029】
(f)細胞培養
正常ヒト表皮細胞の細胞数をK−GM培地にて1×105個に調整し、60mmコラーゲンコートプレート(ファルコン社製)に1mLずつ播種し、K−GM培地を2mLずつ更に各プレートに加えた。95%(V/V)空気−5%(V/V)炭酸ガスの雰囲気下、37℃で4日間静置培養した。培地は2日間毎に交換した。
【0030】
培養上清を吸引除去し、それぞれシモン抽出物を添加したK−GM培地を3mLずつ各ディッシュに加えた。このディッシュを95%(V/V)空気−5%(V/V)炭酸ガスの雰囲気下、37℃で6日間静置培養した。培地は2日間毎に交換した。
【0031】
6日目に培養上清を吸引除去し、0.5mCiの〔14C〕−セリン(アマシャム社製)を培地に添加して、培養を2日間更に行った。培養後、以下のごとく細胞を処理した。
【0032】
(g)脂質抽出
培養上清を吸引除去し、2mLのHepes緩衝液で2回洗浄した後、細胞をスクレイパー(住友ベークライト社製)を用いてディッシュからかきとった。これを1.6mLのHepes緩衝液に懸濁し、4mLのメタノールと2mLのクロロホルムを加え混合した。20分間室温で静置した後、それぞれ1mLのクロロホルムと1.6mLHepes緩衝液を加え混合し、3000回転/分で遠心し下層にあるクロロホルム層を回収し、濃縮遠心によってクロロホルムを取り除き1mLのベンゼンに再溶解した。
【0033】
(h)イアトロビーズカラムを用いたセラミド画分の単離
ベンゼンに溶解した脂質試料を、イアトロビーズ100β1を充填したカラムに供し、ベンゼン−酢酸エチル(4:1)溶液で洗浄した後、1mLの酢酸エチルにて溶出させることにより、セラミド画分を得た。
【0034】
(i)〔14C〕ラベルされたセラミドの放射活性測定
上記セラミド画分に取り込まれた放射活性を液体シンチレーションカウンターにて測定した。
【0035】
試験例1
下記製造例1〜4で調製したシモン抽出物を終濃度0、0.01、0.03、0.1、0.3%のいずれかになるようにヒト表皮細胞培養系に添加した時のセラミド合成能を測定した(表1)。その結果、濃度に依存してヒト表皮細胞のセラミド合成能が促進されることが分かった。
【0036】
【表1】
【0037】
製造例1
シモン(芋)20%エタノール抽出物は、シモン芋乾燥物(熊本製粉社製)100gをミルにて粉砕し、20%エタノール500mLにて1週間抽出を行った。これを濾紙に通し、更に濾過孔径0.22μmの膜(ミリポア社製)で濾過して得た。
【0038】
製造例2
シモン芋乾燥物の代わりにシモン葉乾燥物(熊本製粉社製)100gを用いて抽出した以外は、製造例1と同様の方法によりシモン(葉)20%エタノール抽出物を得た。
【0039】
製造例3
シモン(芋)50%エタノール抽出物は、シモン芋乾燥物100gをミルにて粉砕し、50重量%エタノール250mLにて1週間抽出を行った。これを濾紙に通し、更に濾過孔径0.22μmの膜(ミリポア社製)で濾過して得た。
【0040】
製造例4
シモン芋乾燥物の代わりにシモン葉乾燥物100gを用いて抽出した以外は、製造例3と同様の方法により、シモン(葉)50%エタノール抽出物を得た。
【0041】
実施例14(クリーム)
下記親水性成分を湯浴で80℃に加温し、混合した下記親油性成分に攪拌しながら徐々に加えた。これを、ホモゲナイザーで攪拌して、各成分を充分乳化分散させた後、攪拌しながら徐々に冷却し、シモン芋20%エタノール抽出エキス配合クリームを得た。
【0042】
「親水性成分」
パラオキシ安息香酸メチル 0.1g
プロピレングリコール 6.7g
「親油性成分」
スクワラン 4.7g
白色スクワラン 24.1g
ステアリルアルコール 8.7g
ミリスチン酸イソプロピル 6.0g
モノステアリン酸イソプロピル 1.3g
ポリエチレンアルキルエーテルリン酸 2.3g
モノステアリン酸グリセリン 2.0g
パラオキシ安息香酸 0.1g
製造例1の抽出物 44.1g
【0043】
実施例15〜17(クリーム)
製造例1のシモン(芋)20%エタノール抽出物の代わりに製造例2のシモン(葉)20%エタノール抽出物、製造例3のシモン(芋)50%エタノール抽出物、及び製造例4のシモン(葉)50%エタノール抽出物をそれぞれ用いた以外は、実施例14と同様な方法でそれぞれのクリーム(実施例15〜17)を得た。
【0044】
実施例18(ローション)
製造例1のシモン(芋)20%エタノール抽出物1mLを以下の組成物に加えて常法により100gのローションを得た。
エタノール 10.0g
乳酸 0.3g
クエン酸ナトリウム 0.1g
グリセリン 2.0g
防腐剤、香料及び界面活性剤 適 量
精製水 残 量
【0045】
実施例19〜21(クリーム)
実施例1のシモン(芋)20%エタノール抽出物の代わりに製造例2のシモン(葉)20%エタノール抽出物、製造例3のシモン(芋)50%エタノール抽出物、及び製造例4のシモン(葉)50%エタノール抽出物をそれぞれ用いた以外は、実施例18と同様な方法でそれぞれのクリーム(実施例19〜21)を得た。
【0046】
実施例22〜25(入浴剤)
下記に示す組成で常法により入浴剤を調製した。
【0047】
調製法:各成分を混合し、入浴剤を調製した。尚、この入浴剤は使用時に約3000倍に希釈される。
【0048】
【発明の効果】
以上の如く、本発明により、皮膚表皮層内部で表皮細胞自身のセラミド合成を活発化させ、バリアー機能を改善することによって荒れ肌の改善及び各疾患の治療が期待されるセラミド合成促進剤、皮膚化粧料及び入浴剤を提供できることは明らかである。[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a ceramide synthesis promoter which is expected to improve rough skin and various skin diseases by activating ceramide synthesis of epidermal cells within the skin epidermis layer and improving the skin barrier function. Moreover, it is related with skin cosmetics and a bath agent.
[0002]
[Prior art]
Ceramide, a kind of lipid, has recently been known to have an important physiological role, although it is less in quantity than glycerolipid, which occupies most of the living body. Although it is a physiologically important place in the biodistribution of mammals including humans, it is known that it is accumulated in the brain, liver, skin and the like.
[0003]
In the skin, ceramide accumulates particularly in the stratum corneum, which is synthesized and secreted by epidermal cells and is a major component of intercellular lipids that form a unique lamellar structure between cells (Lukas Landman: Anat. Embryol. 178, 1-3, 1988).
[0004]
The stratum corneum has a series of physiological roles including the moisture retention ability of the skin and physical protection of the living body, the so-called barrier function, but the intercellular lipid is the substance of this barrier function and is the most important in life support. He plays one of the roles (Genkawa Minatogawa: Journal of the Cosmetic Society, 1 (4), 250-253, 1991). From this point of view, it can be said that skin ceramide has become one of the key points of biological defense.
[0005]
In rough skin, dry skin, and various skin diseases, it has been reported that the formation of the stratum corneum is prevented and the barrier function is reduced. Specific examples include a decrease in the turnover of the epidermis layer with aging of the skin surface, or rough skin and dry skin caused by external factors such as light, temperature, and weather conditions. This is thought to be induced as a result of a decrease in the barrier function and a decrease in the water retention capacity inherent in the skin and an increase in the amount of water transpiration (Shuichi Akasaki et al .: Nisshinkai, 98 ( 1), 41-45, 1988).
[0006]
Also, among skin diseases, in atopic dermatitis, barrier function is reduced or collapsed not only in the inflamed part of the patient but also in the non-inflamed part, and the general or specific type of ceramide in the patient's skin. A decrease in content has been reported (Makoto Kawashima: Journal of the Cosmetic Society, 15 (4), 261-262, 1991).
[0007]
In addition, a variation in the amount of ceramide in the patient's skin has also been reported in psoriasis (Strfania Motta et al .: Arch. Dermatol. 130, 452-456, 1994), and this variation is also related to barrier collapse Conceivable.
[0008]
For skin diseases and insufficiency resulting from the reduction or collapse of the skin barrier, conventionally, moisturizing ingredients have been used to prevent skin dryness and moisturize, and anti-inflammatory agents have been used to suppress eczema. Has been. However, since these methods replenish moisture or a moisturizing component on the surface of the stratum corneum, the effect remains temporary, and sufficient moisture cannot be continuously given to the inside of the skin ( Toshiyuki Takemura: Pharmacia, 28, 1, 1992). Even if temporary inflammation was suppressed, there were many problems in sustaining effects and side effects.
[0009]
On the other hand, recently, an attempt was made to improve the skin with external supplementation of ceramide, the main component of the barrier, and it was reported that it was effective for rough skin and atopic dermatitis (Yuko Higaki et al .: Allergy clinical) 13 (12), 26-28, 1993).
[0010]
However, although this method seems to be effective quickly, it is not sufficient in terms of sustainability of the effect like conventional moisturizers, etc. There are also possible disadvantages such as insufficient expression.
[0011]
On the other hand, the effect of the Simon extract on epidermal cells was not particularly known.
[0012]
[Problems to be solved by the invention]
In light of such circumstances, the present inventors intended to activate the ceramide synthesis of the epidermal cells themselves within the skin epidermis layer and improve the skin barrier function, and as a result of intensive studies, the simon extract is effective in synthesizing ceramide. It has been found that it has an accelerating action, and has led to the completion of the present invention, the purpose of which is to improve rough skin by activating ceramide synthesis of the epidermal cells themselves within the skin epidermis layer and various The object is to provide a ceramide synthesis promoter, a skin cosmetic, and a bath agent, which are expected to treat skin diseases.
[0013]
[Means for Solving the Problems]
The above-mentioned object is achieved by a ceramide synthesis accelerator comprising a Simon extract, and a skin cosmetic and a bath containing the Simon extract.
[0014]
DETAILED DESCRIPTION OF THE INVENTION
The Simon used in the present invention is a kind of white sweet potato belonging to the genus Sweet potato family that is cultivated mainly in Central and South America. It is extremely easy to breed and can be easily obtained. Traditionally known to be rich in vitamin K, vitamins and minerals, it is sold as a processed food containing vitamin K / D.
[0015]
The Simon extract used in the present invention can be obtained by extracting with various solvents from the leaves, stems, and roots (rice cakes) of Simon. Moreover, commercially available industrial extract can also be used, for example, industrial Simon leaf extract (made by Kumamoto Flour Mills), industrial Simon extract (Kumamoto Flour Mills), etc. are mentioned.
[0016]
Solvents for obtaining the Simon extract include water, alcohols such as methanol, ethanol and isopropyl alcohol, polyhydric alcohols such as ethylene glycol, propylene glycol and 1,3-butylene glycol, ketones such as acetone, acetic acid Esters such as ethyl, ethers such as diethyl ether, and aromatic compounds such as benzene can be used, and two or more of these can be used in combination.
[0017]
The extraction time may be room temperature extraction or heat extraction, and the extraction time is not limited, but is generally preferably 1 hour to 1 week.
[0018]
In general, Simon is used to extract whole plants or leaves and stems individually, or raw plants as they are or after cutting them. And generally the said extraction solvent 100 parts is used with respect to 5-50 parts of dry matter conversion of Simon.
[0019]
For the Simon extract, the extract may be filtered and the filtrate may be used as it is, but it is usually a concentrated solution obtained by concentrating the filtrate under normal pressure or reduced pressure, or using a solid substance by evaporating the solvent to dryness. It is common to do.
[0020]
The ceramide synthesis promoter according to the present invention can be blended without limitation to those to be ingested, such as cosmetics, quasi-drugs, and pharmaceuticals, as well as beverages and foods.
[0021]
The amount of the Simon extract to be added to the skin cosmetic is 0.001 to 5% by mass (hereinafter referred to as “%”) of the total composition in the case of the Simon leaf ethanol extract based on the total amount of the skin cosmetic. Is preferable), and 0.01 to 0.3% is particularly preferable. In order to exhibit ceramide synthesis more effectively, 0.001% or more is preferable, and 5% or less is preferable in terms of harmony with the color and odor of the composition to be blended. In the cosmetics or bathing agents containing these Simon extracts, other ingredients usually used in pharmaceuticals, quasi drugs, skin cosmetics and bathing agents can be blended simultaneously.
[0022]
The form of the skin cosmetic and bath preparation according to the present invention is not particularly limited. For example, a general skin cosmetic such as an aqueous solution, an alcohol aqueous solution, an emulsion, an ointment, a tablet, a powder, an emulsion, or a lotion. Or it can be set as the form of a bath agent.
[0023]
【Example】
Hereinafter, the present invention will be described in more detail with reference to examples. Prior to the examples, a method for evaluating a ceramide synthesis accelerator will be described.
[0024]
Ceramide synthesis promotion test method (a) Cultured epidermis cells Human normal epidermis keratinocytes used commercially available products (Epidercell / newborn foreskin: manufactured by Kurabo Industries).
[0025]
(B) As a medium for cell culture, MCDB153HAA (purchased from Wako Pure Chemical Industries, Ltd.) is used as a base, hydrocortisone (0.5 μmol / L), ethanolamine (0.1 mmol / L), phosphoethanolamine (0.1 mmol) / L), insulin (5 μg / mL), and K-GM medium supplemented with EGF (epidermal growth factor: 10 ng / mL) were used. At the time of cell growth culture, 4 μL of BPE (bovine pituitary extract, purchased from Kurabo Industries) was added to 1 mL of the medium.
[0026]
(C) Preparation of Hepes buffer solution Hepes (manufactured by Dojindo Laboratories Co., Ltd.) 7.15 g, glucose (manufactured by Kanto Chemical Co., Ltd.) 1.8 g, potassium chloride (manufactured by Kanto Chemical Co., Ltd.) 0.22 g, sodium chloride (Kanto Chemical Co., Ltd.) 7.7 g, disodium hydrogen phosphate dodecahydrate (manufactured by Kanto Chemical Co., Ltd.) 0.27 g dissolved in purified water, adjusted to pH 7.4 with 1 mol / L sodium hydroxide aqueous solution and adjusted to 1 L The female was up.
[0027]
(D) Trypsin solution A Hepes buffer solution containing 0.025% trypsin (manufactured by Sigma) and 0.01% disodium ethylenediaminetetraacetate (manufactured by Kanto Chemical) was prepared.
[0028]
(E) Soybean trypsin inhibitor (hereinafter referred to as SBTI, manufactured by Sigma) was dissolved in Hepes buffer prepared with trypsin reaction stop solution (c) (0.2%) and used as a SBTI solution to stop trypsin reaction. .
[0029]
(F) Cell culture The number of normal human epidermal cells was adjusted to 1 × 10 5 with K-GM medium, seeded on a 60 mm collagen-coated plate (manufactured by Falcon), 1 mL each, and 2 mL each of K-GM medium. Further added to each plate. The culture was statically cultured at 37 ° C. for 4 days in an atmosphere of 95% (V / V) air-5% (V / V) carbon dioxide gas. The medium was changed every 2 days.
[0030]
The culture supernatant was removed by aspiration, and 3 mL of K-GM medium supplemented with Simon extract was added to each dish. The dish was statically cultured at 37 ° C. for 6 days in an atmosphere of 95% (V / V) air-5% (V / V) carbon dioxide gas. The medium was changed every 2 days.
[0031]
On the sixth day, the culture supernatant was removed by aspiration, 0.5 mCi [14C] -serine (manufactured by Amersham) was added to the medium, and the culture was further performed for 2 days. After the culture, the cells were treated as follows.
[0032]
(G) The lipid extraction culture supernatant was removed by aspiration, washed twice with 2 mL of Hepes buffer, and the cells were scraped from the dish using a scraper (manufactured by Sumitomo Bakelite). This was suspended in 1.6 mL of Hepes buffer, and 4 mL of methanol and 2 mL of chloroform were added and mixed. After standing at room temperature for 20 minutes, 1 mL each of chloroform and 1.6 mL Hepes buffer solution were added and mixed, centrifuged at 3000 rpm to recover the lower chloroform layer, and chloroform was removed by concentration centrifugation, and 1 mL of benzene was removed. Redissolved.
[0033]
(H) Isolation of ceramide fraction using iatrobead column Lipid sample dissolved in benzene was applied to a column filled with iatrobead 100β1, washed with benzene-ethyl acetate (4: 1) solution, By eluting with ethyl acetate, a ceramide fraction was obtained.
[0034]
(I) Radioactivity measurement of [ 14 C] -labeled ceramide The radioactivity incorporated into the ceramide fraction was measured with a liquid scintillation counter.
[0035]
Test example 1
When the Simon extract prepared in the following Production Examples 1 to 4 was added to a human epidermal cell culture system so as to have a final concentration of 0, 0.01, 0.03, 0.1, or 0.3%, Ceramide synthesis ability was measured (Table 1). As a result, it was found that the ceramide synthesis ability of human epidermal cells was promoted depending on the concentration.
[0036]
[Table 1]
[0037]
Production Example 1
The 20% ethanol extract of Simmon (salmon) was crushed with 100 g of Simmon Koji dried product (manufactured by Kumamoto Flour Milling Co., Ltd.) and extracted with 500 mL of 20% ethanol for 1 week. This was passed through a filter paper and further filtered through a membrane (manufactured by Millipore) having a filtration pore size of 0.22 μm.
[0038]
Production Example 2
A 20% ethanol extract of Simon (leaves) was obtained in the same manner as in Production Example 1, except that 100 g of dried Simon leaf (manufactured by Kumamoto Flour Milling Co., Ltd.) was used instead of the dried Simmon cake.
[0039]
Production Example 3
Simmon 50% ethanol extract was obtained by pulverizing 100 g of dried Simmon koji using a mill and extracting with 250 mL of 50 wt% ethanol for 1 week. This was passed through a filter paper and further filtered through a membrane (manufactured by Millipore) having a filtration pore size of 0.22 μm.
[0040]
Production Example 4
A Simon (leaf) 50% ethanol extract was obtained in the same manner as in Production Example 3, except that 100 g of dried Simon leaf was used instead of the dried Simmon cake.
[0041]
Example 14 (Cream)
The following hydrophilic component was heated to 80 ° C. in a hot water bath and gradually added to the mixed lipophilic component below with stirring. This was stirred with a homogenizer to sufficiently emulsify and disperse each component, and then gradually cooled with stirring to obtain a cream containing 20% ethanol extract of Simon シ.
[0042]
"Hydrophilic component"
Methyl paraoxybenzoate 0.1g
Propylene glycol 6.7g
"Lipophilic component"
Squalane 4.7g
White squalane 24.1g
8.7 g of stearyl alcohol
Isopropyl myristate 6.0g
Isopropyl monostearate 1.3g
Polyethylene alkyl ether phosphate 2.3g
2.0g glyceryl monostearate
0.1 g paraoxybenzoic acid
44.1 g of extract of Production Example 1
[0043]
Examples 15-17 (cream)
Simon (leaf) 20% ethanol extract of Production Example 2 instead of Simon (rice) 20% ethanol extract of Production Example 1, Simmon 50% ethanol extract of Production Example 3 and Simon of Production Example 4 (Leaf) Each cream (Examples 15 to 17) was obtained in the same manner as in Example 14 except that 50% ethanol extract was used.
[0044]
Example 18 (Lotion)
100 mL lotion was obtained by a conventional method by adding 1 mL of 20% ethanol extract of Simmon (芋) of Production Example 1 to the following composition.
Ethanol 10.0g
Lactic acid 0.3g
Sodium citrate 0.1g
Glycerin 2.0g
Preservatives, perfumes and surfactants Appropriate amount of purified water Residual amount [0045]
Examples 19-21 (cream)
Simmon (leaf) 20% ethanol extract of Production Example 2 instead of Simon (crab) 20% ethanol extract of Example 1 Simmon (Leaf) 50% ethanol extract of Production Example 3 and Simon of Production Example 4 (Leaf) Each cream (Examples 19 to 21) was obtained in the same manner as in Example 18 except that 50% ethanol extract was used.
[0046]
Examples 22-25 (bathing agent)
A bath agent was prepared by a conventional method with the composition shown below.
[0047]
Preparation method: Each component was mixed to prepare a bath agent. In addition, this bath agent is diluted about 3000 times at the time of use.
[0048]
【The invention's effect】
As described above, according to the present invention, the ceramide synthesis promoter, skin makeup, which is expected to improve rough skin and treat various diseases by activating the synthesis of ceramide in the epidermis cells within the skin epidermis layer and improving the barrier function. Obviously, a bath and bathing agent can be provided.
Claims (3)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2000071647A JP3852262B2 (en) | 2000-03-15 | 2000-03-15 | Ceramide synthesis accelerator, skin cosmetic and bath agent |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2000071647A JP3852262B2 (en) | 2000-03-15 | 2000-03-15 | Ceramide synthesis accelerator, skin cosmetic and bath agent |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2001261541A JP2001261541A (en) | 2001-09-26 |
JP3852262B2 true JP3852262B2 (en) | 2006-11-29 |
Family
ID=18590193
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2000071647A Expired - Lifetime JP3852262B2 (en) | 2000-03-15 | 2000-03-15 | Ceramide synthesis accelerator, skin cosmetic and bath agent |
Country Status (1)
Country | Link |
---|---|
JP (1) | JP3852262B2 (en) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2012042970A1 (en) * | 2010-09-28 | 2012-04-05 | 国立大学法人高知大学 | Prophylactic or therapeutic agent for atopic dermatitis, and preparation for external application |
-
2000
- 2000-03-15 JP JP2000071647A patent/JP3852262B2/en not_active Expired - Lifetime
Also Published As
Publication number | Publication date |
---|---|
JP2001261541A (en) | 2001-09-26 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
DE69919287T2 (en) | Ethyl acetate extract of indigo plant | |
JP6688492B2 (en) | Black ginger-containing PPARγ expression promoting composition | |
JP2011088845A (en) | Involucrin expression inhibitor | |
JP2016160198A (en) | Kaempferia parviflora-containing compositions | |
KR102062854B1 (en) | Cosmetic composition containing active fractions of Rubus coreanus MIQ root | |
JP2001163759A (en) | Beautifying and whitening agent, and preparation for external use for skin containing the same | |
JP2004210743A (en) | Agent for promoting production of ceramide | |
KR20170136474A (en) | Cosmetic compositon for moisturizing comprising rehmannia glutinosa, astragalus membranaceus, angelica gigas and scutellaria baicalensis extract as an active ingredient | |
KR101864223B1 (en) | A Composition Of Cosmetics Comprising Sambung Nyawa Extract | |
JP2019147824A (en) | Kaempferia parviflora-containing compositions | |
JP3852262B2 (en) | Ceramide synthesis accelerator, skin cosmetic and bath agent | |
KR102106440B1 (en) | Composition for improving skin condition comprising blueberry and black rice extract fermented lactic acid bacteria | |
JP2000239145A (en) | Anti-allergic agent, beautifully whitening agent and skin cosmetic | |
KR20100132206A (en) | Cosmetic composition for improving skin appearances comprising ginsenoside rh2 as active ingredient | |
JP2021143131A (en) | Anti-aging agent, skin cosmetic and oral composition | |
JP2004155961A (en) | Antioxidant | |
KR102292114B1 (en) | Composition for Improving Skin Conditions Having Moisturizing, Anti-Inflammatory Caused by Fine Dust and Pore Shrinkage Property Comprising Plant Complex Extracts as Active Ingredient | |
JP5155543B2 (en) | Endothelin-1 production inhibitor, hexosaminidase release inhibitor, anti-inflammatory / whitening skin preparation, endothelin-1 production inhibition method, and hexosaminidase release inhibition method | |
JP2005089390A (en) | Cosmetic | |
JP7217550B2 (en) | Composition containing black ginger | |
KR20190033871A (en) | Complex cosmetic composition for improving skin-aging | |
JP2005194239A (en) | Ceramide synthesis promotor and skin care preparation for external use | |
KR20180072381A (en) | Composition for improving skin condition comprising extract of Pisum sativum var. arvense (L.) Poir. | |
KR101924355B1 (en) | Composition for anti-stretch mark effect on the skin containing extracts of salicornia herbacea and ecklonia cava | |
JP2008208053A (en) | Composition for promoting skin cell growth and method for producing the same |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A02 | Decision of refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A02 Effective date: 20040113 |
|
A521 | Written amendment |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20040413 |
|
A711 | Notification of change in applicant |
Free format text: JAPANESE INTERMEDIATE CODE: A711 Effective date: 20040916 |
|
A521 | Written amendment |
Free format text: JAPANESE INTERMEDIATE CODE: A821 Effective date: 20040917 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20060828 |
|
R150 | Certificate of patent or registration of utility model |
Ref document number: 3852262 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20090915 Year of fee payment: 3 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20100915 Year of fee payment: 4 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20100915 Year of fee payment: 4 |
|
S111 | Request for change of ownership or part of ownership |
Free format text: JAPANESE INTERMEDIATE CODE: R313117 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20100915 Year of fee payment: 4 |
|
R350 | Written notification of registration of transfer |
Free format text: JAPANESE INTERMEDIATE CODE: R350 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20100915 Year of fee payment: 4 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20110915 Year of fee payment: 5 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20120915 Year of fee payment: 6 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20130915 Year of fee payment: 7 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
EXPY | Cancellation because of completion of term |