JPH1059839A - Preparation for external use for skin - Google Patents
Preparation for external use for skinInfo
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- JPH1059839A JPH1059839A JP8239702A JP23970296A JPH1059839A JP H1059839 A JPH1059839 A JP H1059839A JP 8239702 A JP8239702 A JP 8239702A JP 23970296 A JP23970296 A JP 23970296A JP H1059839 A JPH1059839 A JP H1059839A
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- Prior art keywords
- skin
- glutathione
- external preparation
- oxidized
- salts
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- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
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- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
Abstract
Description
【０００１】[0001]
【産業上の利用分野】この発明は、皮膚の色黒，シミ，
ソバカス、或いは日焼けによる色素沈着を改善し、また
抗酸化作用及びこれに基づく皮膚の老化を防止又は予防
し、かつ皮膚刺激性が低く安全でさらに安定である皮膚
外用剤に関する。さらに詳しくは、酸化型及び還元型グ
ルタチオンのアルキルエステルとアミノ酸との塩類から
選ばれた１種又は２種以上を配合してなる皮膚外用剤、
或いは酸化型グルタチオンのアルキルエステルとアミノ
酸との塩類から選ばれた１種又は２種以上と還元型ニコ
チンアミドアデニンジヌクレオチド（ＮＡＤＨ）及び／
又は還元型ニコチンアミドアデニンジヌクレオチドリン
酸（ＮＡＤＰＨ）とを併用してなる皮膚外用剤に関す
る。BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to the use of black and white skin,
The present invention relates to an external preparation for skin which improves pigmentation due to freckles or sunburn, prevents or prevents an antioxidant effect and skin aging based on the antioxidant effect, and has low skin irritation and is safe and more stable. More specifically, a skin external preparation comprising one or more selected from salts of alkyl esters of oxidized and reduced glutathione and amino acids,
Alternatively, one or more selected from salts of an alkyl ester of oxidized glutathione and an amino acid and reduced nicotinamide adenine dinucleotide (NADH) and / or
Or, it relates to an external preparation for skin which is used in combination with reduced nicotinamide adenine dinucleotide phosphate (NADPH).
【０００２】[0002]
【従来の技術】還元型グルタチオンは、生体内の酸化還
元系に関与し生体内で活性酸素種を捕捉する抗酸化作
用、ＳＨ酵素などいろいろの酵素を活性化すると共に、
補酵素としても作用すること、またメルカプツール酸抱
合等の解毒反応にも関与し、さらにチロシナーゼの活性
及び生合成を阻害することが知られている。そのため、
還元型グルタチオンを配合した皮膚外用剤は、老化防止
作用と美白作用を合わせ持つ有効な製剤として期待され
ている。2. Description of the Related Art Reduced glutathione is involved in a redox system in a living body and activates various enzymes such as an antioxidant action to capture reactive oxygen species in the living body and an SH enzyme.
It is known to act as a coenzyme, to participate in detoxification reactions such as mercapturic acid conjugation, and to inhibit tyrosinase activity and biosynthesis. for that reason,
Skin external preparations containing reduced glutathione are expected to be effective preparations having both an antiaging effect and a whitening effect.
【０００３】また、酸化型グルタチオンは、生体内でＮ
ＡＤＨやＮＡＤＰＨを水素供与体として、グルタチオン
リダクターゼの作用を受けて還元型グルタチオンに変化
し、生理活性を発揮することが知られている。[0003] In addition, oxidized glutathione is used in vivo to produce N
It is known that ADH or NADPH is used as a hydrogen donor to change into reduced glutathione under the action of glutathione reductase, thereby exhibiting a physiological activity.
【０００４】しかしながら、酸化型或いは還元型グルタ
チオン（以下グルタチオン類と略す）は、水溶性の性質
を有しているために、経皮透過性及び細胞膜透過性が低
く、有効な美白作用及び抗酸化作用を発現させるために
はかなり高濃度を外用剤基剤中に配合する必要があっ
た。そこで、グルタチオン類の分子中のカルボキシル基
を低級アルコールでアルキルエステル化することにより
脂溶性を付与し、配合した皮膚化粧料が提案されている
（特公昭４８−１５０５）。[0004] However, oxidized or reduced glutathione (hereinafter abbreviated as glutathione) has a low water-soluble property and therefore has low transdermal permeability and low cell membrane permeability, and is effective in whitening and antioxidant. In order to exert the effect, it was necessary to mix a considerably high concentration in the external preparation base. Therefore, there has been proposed a skin cosmetic composition in which a carboxyl group in the molecule of glutathione is alkyl-esterified with a lower alcohol to impart lipophilicity and blended (Japanese Patent Publication No. 48-1505).
【０００５】[0005]
【発明が解決しようとする課題】本発明は、皮膚刺激性
や副作用の問題がなく、安全且つ安定で、非常に優れた
美白効果及び老化防止効果を発揮する皮膚外用剤を提供
することを目的とする。すなわち、美白効果及び老化防
止効果を有するグルタチオン類の経皮及び細胞膜透過性
を高めて、表皮基底層に有効に到達させることを可能と
するものである。SUMMARY OF THE INVENTION An object of the present invention is to provide a skin external preparation which is safe and stable, has no problem of skin irritation and side effects, and has an extremely excellent whitening effect and anti-aging effect. And That is, the transdermal and cell membrane permeability of glutathione having a whitening effect and an anti-aging effect is enhanced, and it is possible to effectively reach the basal layer of the epidermis.
【０００６】[0006]
【課題を解決するための手段】従って本発明において
は、グルタチオン類の経皮及び細胞膜透過性を高めるた
め、グルタチオン類誘導体の検討を行った。その結果、
酸化型及び還元型のグルタチオンのアルキルエステルと
アミノ酸との塩類を配合することにより、これらの経皮
及び細胞膜透過性は顕著に高まり、同一配合量における
チロシナーゼ活性或いは生合成阻害作用及び抗酸化作用
が驚くほど向上することを見出した。従って皮膚外用剤
基剤に対する前記グルタチオン類のエステルの配合量は
少量でよく、外用剤の製剤安定性に及ぼす影響も小さ
く、さらに皮膚刺激の発生等安全性上の問題もない。Accordingly, in the present invention, glutathione derivatives have been studied in order to increase the transdermal and cell membrane permeability of glutathiones. as a result,
By blending salts of oxidized and reduced glutathione alkyl esters with amino acids, their percutaneous and cell membrane permeability are remarkably enhanced, and tyrosinase activity or biosynthesis inhibitory action and antioxidant action at the same amount are reduced. I found that it improved surprisingly. Therefore, the glutathione ester may be incorporated in a small amount with respect to the skin external preparation base, the effect of the external preparation on the preparation stability is small, and there is no safety problem such as generation of skin irritation.
【０００７】本発明で用いられる、酸化型及び還元型の
グルタチオンのアルキルエステルとアミノ酸との塩類
は、式（１）及び式（２）で表される化合物で、The oxidized and reduced glutathione alkyl esters and amino acids used in the present invention are compounds represented by formulas (1) and (2):
【０００８】[0008]
【化１】 Embedded image
【化２】 ＡＡは、上記エステル類と塩を形成するアミノ酸であ
り、Ｒ1は炭素数１〜１２のアルキル基、Ｒ2は水素基若
しくは炭素数１〜１２のアルキル基である。Embedded image AA is an amino acid that forms a salt with the above esters, R1 is an alkyl group having 1 to 12 carbon atoms, and R2 is a hydrogen group or an alkyl group having 1 to 12 carbon atoms.
【０００９】エステル類と塩を形成するアミノ酸は、水
溶性及び生理学的相溶性の面からグリシン，アラニン，
アスパラギン酸及びグルタミン酸のような天然のα−ア
ミノ酸、ピログルタミン酸，γ−アミノ酪酸及びタウリ
ンが例示される。Amino acids that form salts with esters are glycine, alanine, and glycine from the viewpoint of water solubility and physiological compatibility.
Examples are natural α-amino acids such as aspartic acid and glutamic acid, pyroglutamic acid, γ-aminobutyric acid and taurine.
【００１０】また、アルキルエステルを形成するアルキ
ル基の炭素数は、１〜１２が好ましく、水溶性及び脂溶
性の面から１〜６が特に好ましい。またアルキル基若し
くはアルケニル基は直鎖状であっても分岐を有してもか
まわない。The number of carbon atoms of the alkyl group forming the alkyl ester is preferably from 1 to 12, and particularly preferably from 1 to 6 from the viewpoint of water solubility and fat solubility. The alkyl group or alkenyl group may be linear or branched.
【００１１】本発明で用いられる、酸化型及び還元型の
グルタチオンのアルキルエステルとアミノ酸との塩類
は、例えば特表平６−５０９８１７に記載された方法で
製造することができる。すなわち、グルタチオンアルキ
ルエステルを水中でアミノ酸で処理した後、減圧下で溶
媒を除去し、得られた反応生成物をアルコールで沈殿さ
せて単離することによって製造することができる。この
ようにして得られた塩は、そのままでも、冷エタノール
又は他の有機溶媒で洗浄して用いてもよい。さらに、こ
の塩において、結晶水若しくはエタノールを含んでもよ
い。The oxidized and reduced glutathione alkyl esters and amino acid salts used in the present invention can be produced, for example, by the method described in JP-A-6-509817. That is, it can be produced by treating a glutathione alkyl ester with an amino acid in water, removing the solvent under reduced pressure, and precipitating and isolating the obtained reaction product with alcohol. The salt thus obtained may be used as it is, or washed with cold ethanol or another organic solvent. Further, the salt may contain water of crystallization or ethanol.
【００１２】[0012]
【発明の実施の形態】上記の酸化型及び還元型のグルタ
チオンのアルキルエステルとアミノ酸との塩類を通常の
皮膚外用剤基剤中に配合して皮膚外用剤とする。配合量
は、製剤中の有効濃度や製剤の安定性等を考慮して０.
００１〜１０重量％程度が適当である。外用剤の形態と
しては、ローション，乳剤，クリーム，軟膏等、種々の
形態をとることができる。また、化粧水，美容液，乳
液，クリーム等の老化防止用及び美白用化粧料としても
提供することができる。BEST MODE FOR CARRYING OUT THE INVENTION Salts of the above-mentioned oxidized and reduced alkyl glutathione esters and amino acids are blended into an ordinary skin external preparation base to prepare a skin external preparation. The amount to be blended is set at 0.1 considering the effective concentration in the preparation and the stability of the preparation.
About 001 to 10% by weight is appropriate. The external preparation may take various forms such as lotions, emulsions, creams, ointments and the like. In addition, it can be provided as an anti-aging and whitening cosmetic such as lotion, serum, milky lotion, cream and the like.
【００１３】また、酸化型グルタチオンのエステルを配
合した場合は、水素供与体であるＮＡＤＨ及び／又はＮ
ＡＤＰＨと併用することにより、前記の生理活性がさら
に高められることを見出した。When an oxidized glutathione ester is blended, the hydrogen donors NADH and / or N
It has been found that the use of ADPH in combination further enhances the aforementioned physiological activity.
【００１４】[0014]
【作用】本発明で用いる酸化型及び還元型のグルタチオ
ンのアルキルエステルとアミノ酸との塩類のメラニン色
素白色化作用、チロシナーゼ活性阻害作用及びチロシナ
ーゼ生合成阻害作用について以下に示す。作用を測定し
た化合物を表１にまとめて示した。比較のため、酸化型
グルタチオン及び還元型グルタチオンを用いて同様に作
用の測定を行った。The whitening action, tyrosinase activity inhibitory action and tyrosinase biosynthesis inhibitory action of salts of oxidized and reduced glutathione alkyl esters and amino acids used in the present invention are shown below. The compounds whose effects were measured are summarized in Table 1. For comparison, the effect was similarly measured using oxidized glutathione and reduced glutathione.
【００１５】[0015]
【表１】 [Table 1]
【００１６】メラニン色素白色化作用は、グルタチオン
類を精製水に溶解した後、適当量の培地にて希釈して試
料とした。マウスＢ16メラノーマ細胞の懸濁液（細胞数
５０,０００程度）に添加し、３日間培養を行った後、
細胞を分離し、測色して評価した。グルタチオン類を添
加せず同様に処理した系を対照としてＬ値の差を求め、
図１に示した。グルタチオンを添加した系と比較して、
グルタチオンアルキルエステルのアミノ酸との塩類を添
加した系では、Ｌ値の差が３倍以上大きくなっており、
グルタチオンアルキルエステルをアミノ酸塩として配合
することにより、細胞膜透過性が向上しメラニン色素白
色化作用が顕著に上昇した。For the melanin pigment whitening effect, a sample was prepared by dissolving glutathione in purified water and diluting it with an appropriate amount of medium. After adding to a suspension of mouse B16 melanoma cells (about 50,000 cells) and culturing for 3 days,
Cells were separated, colorimetrically evaluated. The difference in L value was determined using a system treated in the same manner without adding glutathione as a control,
As shown in FIG. Compared to the system to which glutathione was added,
In a system to which a salt of glutathione alkyl ester with an amino acid is added, the difference in L value is three times or more,
By mixing the glutathione alkyl ester as an amino acid salt, the cell membrane permeability was improved and the melanin pigment whitening effect was significantly increased.
【００１７】チロシナーゼ活性阻害作用は、グルタチオ
ン類を精製水に溶解して試料とし、酵素溶液としてチロ
シナーゼ（５０，０００units，シグマ社製）を精製水
で６００倍希釈したものを用い、チロシン５０ｍｇを精
製水１００ｍｌに溶解して基質溶液として評価した。グ
ルタチオン類は、最終濃度が１．０μＭ〜１００ｍＭの
範囲で段階的に変化するように添加した。The tyrosinase activity inhibitory effect is obtained by dissolving glutathione in purified water to prepare a sample, and using a tyrosinase (50,000 units, Sigma) diluted 600 times with purified water as an enzyme solution to purify 50 mg of tyrosine. It was dissolved in 100 ml of water and evaluated as a substrate solution. Glutathiones were added so that the final concentration was changed stepwise in the range of 1.0 μM to 100 mM.
【００１８】酵素反応は、試料溶液２ｍｌ，１／１５Ｍ
リン酸緩衝液（ｐＨ６．８）２ｍｌ，基質溶液０．５ｍ
ｌ，酵素溶液０．５ｍｌを混合し、３７℃にて１時間イ
ンキュベートして行わせた。反応後４７５ｎｍにおける
吸光度（Ａs）を測定した。さらに試料溶液の代わりに
精製水を添加した系，及び基質溶液の代わりに精製水を
添加した系において同様に反応させ、それぞれの場合に
おける吸光度（Ａb及びＡ0）を測定した。チロシナーゼ
活性阻害率は数式(1)により求めた。The enzymatic reaction is carried out in a sample solution 2 ml, 1 / 15M
2 ml of phosphate buffer (pH 6.8), substrate solution 0.5 m
1, 0.5 ml of enzyme solution was mixed and incubated at 37 ° C. for 1 hour. After the reaction, the absorbance (As) at 475 nm was measured. Further, the same reaction was performed in a system in which purified water was added instead of the sample solution and in a system in which purified water was added instead of the substrate solution, and the absorbance (Ab and A0) in each case was measured. The tyrosinase activity inhibition rate was determined by equation (1).
【００１９】[0019]
【数１】 (Equation 1)
【００２０】チロシナーゼ生合成阻害作用の評価は次の
ようにして行った。グルタチオン類の水溶液をマウスＢ
16メラノーマ細胞（細胞数５０，０００程度）に添加
し、３日間培養を行った後、以下の方法により細胞中の
チロシナーゼ活性を測定した。すなわち、１／１５Ｍリ
ン酸緩衝液（ｐＨ６．８）２ｍｌに１．０重量％のドー
パ水溶液０．５ｍｌ及び培養細胞液０．５ｍｌを混合
し、３７℃にて１時間インキュベートした後、４０５ｎ
ｍにおける吸光度（Ａs）を測定した。対照として溶媒
の精製水のみを同様に添加して培養し、同様に培養細胞
液とドーパ水溶液とをインキュベートして吸光度（Ａ
b）を測定し、数式(2)によりチロシナーゼ生合成阻害率
を求めた。グルタチオン類は、最終濃度が１．０μＭ〜
１００ｍＭの範囲で段階的に変化するように添加した。The tyrosinase biosynthesis inhibitory activity was evaluated as follows. Mouse B with aqueous solution of glutathione
After adding the cells to 16 melanoma cells (about 50,000 cells) and culturing for 3 days, the tyrosinase activity in the cells was measured by the following method. That is, 0.5 ml of a 1.0% by weight dopa aqueous solution and 0.5 ml of a cultured cell solution were mixed with 2 ml of a 1/15 M phosphate buffer (pH 6.8), and incubated at 37 ° C. for 1 hour.
The absorbance (As) at m was measured. As a control, culture was performed by adding only purified water as a solvent in the same manner, and the cultured cell solution and the dopa aqueous solution were similarly incubated, and the absorbance (A
b) was measured, and the tyrosinase biosynthesis inhibition rate was determined by equation (2). Glutathiones have a final concentration of 1.0 μM or more.
It was added so as to change stepwise in the range of 100 mM.
【００２１】[0021]
【数２】 (Equation 2)
【００２２】各試料について、ドーズレスポンス曲線を
作成し、５０％阻害濃度（ＩＤ50）を求めて、表２にま
とめて示した。表２において、酸化型及び還元型のグル
タチオンのアルキルエステルとアミノ酸との塩類のチロ
シナーゼ活性阻害作用及びチロシナーゼ生合成阻害作用
についてのＩＤ50は、エステル化していないグルタチオ
ン類の場合の１／２１１から１／６２及び１／８５から
１／２９となり、酸化型及び還元型のグルタチオンのア
ルキルエステルとアミノ酸との塩類を添加することによ
り、同一濃度で添加した場合、チロシナーゼ活性阻害作
用及びチロシナーゼ生合成阻害作用は大幅に向上する。For each sample, a dose response curve was prepared, and the 50% inhibitory concentration (ID 50 ) was determined. The results are shown in Table 2. In Table 2, the ID 50 for the tyrosinase activity inhibitory activity and tyrosinase biosynthesis inhibitory activity of salts of alkyl and amino acids of oxidized and reduced glutathione is from 1/211 to 1 in the case of non-esterified glutathione. / 62 and 1/85 to 1/29, and by adding salts of oxidized and reduced forms of glutathione alkyl ester and amino acid, when added at the same concentration, tyrosinase activity inhibitory action and tyrosinase biosynthesis inhibitory action Is greatly improved.
【００２３】[0023]
【表２】 [Table 2]
【００２４】酸化型及び還元型のグルタチオンのアルキ
ルエステルとアミノ酸との塩類の活性酸素種による細胞
傷害防御作用は、ヒト線維芽細胞に活性酸素種を曝露
し、その際の細胞生存率を指標として評価できる。培養
したヒト線維芽細胞に、グルタチオン類のエステルを作
用させた後、洗浄し、ヒポキサンチン-キサンチンオキ
シダーゼ系にて活性酸素種（スーパーオキシド及びヒド
ロキシラジカル）を曝露した。曝露後の細胞生存率はＭ
ＴＴ還元法により測定した。[0024] The cytotoxic effect of salts of oxidized and reduced forms of glutathione alkyl esters and amino acids by reactive oxygen species is determined by exposing human fibroblasts to reactive oxygen species and using the cell viability at that time as an index. Can be evaluated. Glutathione esters were allowed to act on the cultured human fibroblasts, washed, and exposed to reactive oxygen species (superoxide and hydroxyl radical) using a hypoxanthine-xanthine oxidase system. Cell viability after exposure is M
It was measured by the TT reduction method.
【００２５】なおＭＴＴ還元法は、細胞のミトコンドリ
ア内に存在する脱水素酵素が基質に作用して生じるＮＡ
ＤＨにより、系に添加したＭＴＴ（2-(4,5-ジメチル−2
-チアゾリル)-3,5-ジフェニルテトラゾリウム ブロミ
ド）のテトラゾリウム環が開環することを利用する測定
方法である。テトラゾリウム環の開環により青色のフォ
ルマザンが生成するので、これを５６０ｎｍにおける吸
光度測定により定量する。In the MTT reduction method, the dehydrogenase present in the mitochondria of a cell acts on a substrate to generate NA.
With DH, MTT (2- (4,5-dimethyl-2) added to the system was added.
-Thiazolyl) -3,5-diphenyltetrazolium bromide). Since blue formazan is generated by the opening of the tetrazolium ring, this is quantified by measuring absorbance at 560 nm.
【００２６】その結果、図２に示すように、酸化型及び
還元型のグルタチオンのアルキルエステルとアミノ酸と
の塩類で処理した場合は、７０〜８５％の良好な細胞生
存率を示した。これに対し、エステル塩化していないグ
ルタチオン類で処理した場合は、３０％未満の低い細胞
生存率を示した。As a result, as shown in FIG. 2, when treated with a salt of an oxidized or reduced glutathione alkyl ester and an amino acid, a good cell viability of 70 to 85% was exhibited. In contrast, when treated with glutathiones that were not esterified, a low cell viability of less than 30% was exhibited.
【００２７】[0027]
【実施例】さらに、本発明について実施例により詳細に
説明する。EXAMPLES Further, the present invention will be described in more detail with reference to examples.
【００２８】 実施例 乳液 (１) スクワラン ５．０（重量％) (２) 白色ワセリン ２．０ (３) ミツロウ ０．５ (４) ソルビタンセスキオレエート ０．８ (５) ポリオキシエチレンオレイルエーテル(20EO) １．２ (６) 1,3-ブチレングリコール ５．０ (７) 酸化型及び還元型のグルタチオンのアルキル 若しくはアルケニルエステルとアミノ酸との塩類 １．０ 及びＮＡＤＰＨ，ＮＡＤＨ (８) 精製水 ５９．０ (９) カルボキシビニルポリマー1.0重量％水溶液 ２０．０ (10) 水酸化カリウム ０．１ (11) エタノール ５．０ (12) パラオキシ安息香酸メチル ０．２ (13) 香料 ０．２ (１)〜(５)の油相成分を混合し７５℃に加熱して溶解，
均一化する。一方(６)〜(８)の水相成分を混合，溶解し
て７５℃に加熱し、前記の油相成分を添加して予備乳化
する。(９)を添加した後ホモミキサーにて均一に乳化
し、(10)を加えてｐＨを調整する。冷却後４０℃にて、
(12)，(13)の成分を(10)に溶解して添加，混合，均一化
する。Example Emulsion (1) Squalane 5.0 (% by weight) (2) White petrolatum 2.0 (3) Beeswax 0.5 (4) Sorbitan sesquioleate 0.8 (5) Polyoxyethylene oleyl ether (20EO) 1.2 (6) 1,3-butylene glycol 5.0 (7) Oxidized and reduced alkyl or alkenyl esters of glutathione and salts with amino acids 1.0 and NADPH, NADH (8) Purified water 59.0 (9) 1.0% by weight aqueous solution of carboxyvinyl polymer 20.0 (10) Potassium hydroxide 0.1 (11) Ethanol 5.0 (12) Methyl parahydroxybenzoate 0.2 (13) Fragrance 0.2 ( The oil phase components of 1) to (5) are mixed and heated to 75 ° C. to dissolve,
Make uniform. On the other hand, the aqueous phase components (6) to (8) are mixed and dissolved, heated to 75 ° C., and the oil phase component is added and pre-emulsified. After adding (9), the mixture is emulsified uniformly with a homomixer, and (10) is added to adjust the pH. At 40 ° C after cooling,
The components (12) and (13) are dissolved in (10), added, mixed and homogenized.
【００２９】上記乳液に配合した酸化型及び還元型のグ
ルタチオンのアルキルエステルとアミノ酸との塩類を、
表３に示す。また還元型グルタチオン及び酸化型グルタ
チオンとＮＡＤＰＨ或いはＮＡＤＨを配合したものを比
較例とした。The salts of the oxidized and reduced glutathione alkyl esters and amino acids blended in the emulsion are
It is shown in Table 3. Further, a mixture of reduced glutathione and oxidized glutathione with NADPH or NADH was used as a comparative example.
【００３０】[0030]
【表３】 [Table 3]
【００３１】皮膚の老化防止効果（しわ発生防止効果） ヘアレスマウス５匹を一群とし、各群について本発明の
実施例及び比較例をそれぞれ１日１回背部に塗布し、１
Ｊ／平方ｃｍ／週のＵＶＡを５０週間照射し、しわの発
生状況を経時的に肉眼観察により評価した。しわの発生
状況は、「発生せず；０点」，「微小なしわがわずかに
発生；１点」，「軽微なしわが明確に発生；２点」，
「中程度のしわが発生；３点」，「深いしわが発生；４
点」として点数化し、各群の平均点を算出した。その
際、精製水を塗布した群を対照とした。Skin Aging Prevention Effect (Wrinkle Prevention Effect) Five hairless mice were grouped as a group, and the Examples and Comparative Examples of the present invention were applied to the back once a day for each group.
UVA of J / square cm / week was irradiated for 50 weeks, and the occurrence of wrinkles was evaluated by visual observation over time. The wrinkle occurrence status is “No occurrence; 0 point”, “Small wrinkle occurs; 1 point”, “Minor wrinkle clearly occurs; 2 points”,
"Medium wrinkles occurred; 3 points", "Deep wrinkles occurred; 4
The points were scored as "points", and the average score of each group was calculated. At that time, a group to which purified water was applied was used as a control.
【００３２】使用試験 皮膚の弾性の低下及びシミ，ソバカス，日焼け等の色素
沈着を主な症状として有するパネラー２０名を一群と
し、各群にそれぞれ実施例及び比較例をブラインドにて
顔面及び手に使用させ、皮膚弾性及び色素沈着の変化を
観察し、評価した。使用期間は４月から１０月の６カ月
間とした。皮膚弾性については、「上昇」，「やや上
昇」，「変化なし」，「低下」の４段階で、美白効果に
ついては、「改善」，「やや改善」，「変化なし」の３
段階にて評価をし、各評価を得たパネラー数にて結果を
示した。Usage Test A group of 20 panelists, whose main symptoms were decreased skin elasticity and pigmentation such as spots, freckles, and sunburn, were placed in a group, and the examples and comparative examples were blindly applied to the face and hands in each group. After use, changes in skin elasticity and pigmentation were observed and evaluated. The period of use was 6 months from April to October. The skin elasticity has four levels of "rise", "slight increase", "no change" and "decrease", and the whitening effect has three levels of "improvement", "slight improvement" and "no change".
The evaluation was performed at each stage, and the results were shown by the number of panelists who obtained each evaluation.
【００３３】[0033]
【表４】 [Table 4]
【００３４】皮膚の老化防止効果の結果を表４に示し
た。老化防止効果測定結果より、酸化型及び還元型のグ
ルタチオンのアルキルエステルとアミノ酸との塩類を配
合した実施例においては、いずれもしわの発生が顕著に
抑制されており、ＵＶＡを５０週間照射した後において
も、各群とも微小なしわの発生を認めただけであった。
これに対してエステル塩としないグルタチオン類を配合
した比較例３〜６においては、対照に比べ若干のしわ発
生抑制効果は認められるが、いずれにおいても明確なし
わの形成が見られ、形成されたしわはほぼ中程度の深さ
にまで達していた。Table 4 shows the results of the skin anti-aging effect. From the measurement results of the anti-aging effect, in Examples in which the salts of the oxidized and reduced forms of glutathione alkyl ester and amino acid were blended, the occurrence of wrinkles was significantly suppressed, and after irradiation with UVA for 50 weeks, In each group, only wrinkles were observed in each group.
On the other hand, in Comparative Examples 3 to 6 in which glutathiones were not used as ester salts, a slight wrinkle generation suppressing effect was observed as compared with the control, but clear wrinkles were formed and formed in all cases. The wrinkles had reached almost medium depth.
【００３５】[0035]
【表５】 [Table 5]
【００３６】表５に示した使用試験結果から明らかなよ
うに、酸化型及び還元型のグルタチオンのアルキルエス
テルとアミノ酸との塩類を配合した実施例を使用したパ
ネラーは全員皮膚弾性の上昇及び色素沈着の改善が認め
られた。これに対し、比較例３〜６を使用したパネラー
は、弾性がやや上昇したパネラーもいるが、明確に上昇
の認められたパネラーはおらず、美白効果に関しても、
はっきりと改善が認められたパネラーはおらず、４０〜
５０％のパネラーで変化を認めなかった。以上の結果よ
り、酸化型及び還元型のグルタチオンのアルキルエステ
ルとアミノ酸との塩類を配合することにより、酸化型グ
ルタチオン及び還元型グルタチオンを配合した場合と比
較して、老化防止効果及び美白効果が飛躍的に向上して
いることがわかる。As is clear from the results of the use test shown in Table 5, all the panelers using the examples in which the salts of the oxidized and reduced forms of glutathione alkyl ester and the amino acid were blended showed an increase in skin elasticity and pigmentation. Improvement was observed. On the other hand, the panelers using Comparative Examples 3 to 6 had some panels whose elasticity was slightly increased, but none of the panelers were clearly observed to have an increased elasticity.
None of the panelists clearly improved, and
No change was observed in 50% of the panelists. From the above results, the antioxidant effect and the whitening effect are remarkably improved by mixing the salts of the oxidized and reduced glutathione alkyl esters and the amino acids as compared with the case where the oxidized glutathione and the reduced glutathione are mixed. It can be seen that the improvement has been achieved.
【００３７】なお、上記の使用期間において、いずれの
実施例を使用した群においても、痛み、痒み等の皮膚刺
激やアレルギー反応等の皮膚症状を訴えたパネラーはい
なかった。また、乳化状態の悪化や配合成分の沈降，変
質等も認められなかった。During the above-mentioned period of use, none of the groups using any of the examples reported any skin irritation such as pain or itchiness or skin symptoms such as allergic reaction. In addition, no deterioration of the emulsified state, no sedimentation and deterioration of the components were observed.
【００３８】 実施例８ 液状皮膚外用剤 (１)グリセリン ２０．０(重量％) (２)プロピレングリコール １０．０ (３)ポリオキシエチレン(５ＥＯ)硬化ヒマシ油 ２．０ (４)精製水 ６１．７ (５)エタノール ５．０ (６)パラオキシ安息香酸メチル ０．３ (７)還元型グルタチオンエチルエステルタウリン塩 １．０ (１)〜(７)の成分を順次添加し、混合，均一化する。Example 8 Liquid Skin External Preparation (1) Glycerin 20.0 (% by weight) (2) Propylene glycol 10.0 (3) Polyoxyethylene (5EO) hydrogenated castor oil 2.0 (4) Purified water 61 0.7 (5) Ethanol 5.0 (6) Methyl paraoxybenzoate 0.3 (7) Reduced glutathione ethyl ester taurine salt 1.0 The components (1) to (7) are sequentially added, mixed and homogenized. I do.
【００３９】 実施例９ 美容液 (１)グリセリン ５．０(重量％) (２)プロピレングリコール ４．０ (３)還元型グルタチオンメチルエステル １．０ γ−アミノ酪酸塩 (４)精製水 ７９．９ (５)エタノール １０．０ (６)パラオキシ安息香酸メチル ０．１ (１)〜(６)の成分を順次添加し、混合，均一化する。Example 9 Serum (1) Glycerin 5.0 (% by weight) (2) Propylene glycol 4.0 (3) Reduced glutathione methyl ester 1.0 γ-aminobutyrate (4) Purified water 9 (5) Ethanol 10.0 (6) Methyl parahydroxybenzoate 0.1 The components (1) to (6) are sequentially added, mixed and homogenized.
【００４０】 実施例１０ Ｗ／Ｏ乳化型クリーム (１)ミツロウ ３．０(重量％) (２)吸着精製ラノリン ５．０ (３)スクワラン ２０．０ (４)固形パラフィン ２．０ (５)マイクロクリスタリンワックス ５．０ (６)アジピン酸ヘキシルデシル １０．０ (７)セスキオレイン酸ソルビタン ３．５ (８)ポリオキシエチレン(５０ＥＯ)硬化ヒマシ油 １．０ (９)１，３−ブチレングリコール ５．０ (10)還元型グルタチオンエチルエステルピログルタミン酸塩 １．０ (11)精製水 ３９．３ (12)パラオキシ安息香酸メチル ０．２ (13)エタノール ５．０ (1)〜(8)の油相成分を混合し７５℃に加熱して溶解，均
一化する。一方(9)〜(11)の水相成分を混合，溶解して
７５℃に加熱し、前記の油相成分に添加してホモミキサ
ーにて均一に乳化する。冷却後４０℃にて、(12)の成分
を(13)に溶解して添加，混合，均一化する。Example 10 W / O emulsified cream (1) Beeswax 3.0 (% by weight) (2) Adsorbed and purified lanolin 5.0 (3) Squalane 20.0 (4) Solid paraffin 2.0 (5) Microcrystalline wax 5.0 (6) Hexyldecyl adipate 10.0 (7) Sorbitan sesquioleate 3.5 (8) Polyoxyethylene (50EO) hydrogenated castor oil 1.0 (9) 1,3-butylene glycol 5.0 (10) Reduced glutathione ethyl ester pyroglutamate 1.0 (11) Purified water 39.3 (12) Methyl paraoxybenzoate 0.2 (13) Ethanol 5.0 (1) to (8) The oil phase components are mixed and heated to 75 ° C. to dissolve and homogenize. On the other hand, the aqueous phase components (9) to (11) are mixed and dissolved, heated to 75 ° C., added to the above oil phase components, and uniformly emulsified by a homomixer. After cooling, at 40 ° C., the component (12) is dissolved in (13), added, mixed and homogenized.
【００４１】 実施例 Ｏ／Ｗ型乳剤性軟膏 (１)白色ワセリン ２５．０(重量％) (２)ステアリルアルコール １５．０ (３)ラウリル硫酸ナトリウム １．０ (４)パラオキシ安息香酸ブチル ０．３ (５)精製水 ５７．７ (６)酸化型グルタチオンイソプロピルエステル ０．９５ アスパラギン酸塩 (７)ＮＡＤＰＨ ０．０５ (１)〜(４)の油相成分を混合し７５℃に加熱して溶解，
均一化する。７５℃に加熱した(５)に油相成分を添加し
て乳化する。冷却後４０℃にて、(６)，(７)の成分を添
加，混合，均一化する。Example O / W Type Emulsion Ointment (1) White Vaseline 25.0 (% by weight) (2) Stearyl Alcohol 15.0 (3) Sodium Lauryl Sulfate 1.0 (4) Butyl Paraoxybenzoate 0. 3 (5) Purified water 57.7 (6) Oxidized glutathione isopropyl ester 0.95 aspartate (7) NADPH 0.05 Mix the oil phase components of (1) to (4) and heat to 75 ° C. Dissolution,
Make uniform. The oil phase component is added to (5) heated to 75 ° C. and emulsified. After cooling, the components (6) and (7) are added, mixed and homogenized at 40 ° C.
【００４２】[0042]
【発明の効果】以上詳述したように、本発明により、少
量の酸化型及び還元型のグルタチオンのアルキルエステ
ルとアミノ酸との塩類を配合することにより、非常に優
れた美白効果及び老化防止効果を発揮する皮膚外用剤を
提供することができる。本発明に係る皮膚外用剤におい
ては、有効成分であるグルタチオン類が良好に表皮，真
皮及び表皮基底層のメラノサイトに到達でき、低濃度の
配合で優れた老化防止効果及び美白効果を示す。従っ
て、皮膚刺激性や感作性の発現するおそれがない。As described in detail above, according to the present invention, by adding a small amount of an oxidized or reduced glutathione alkyl ester and a salt of an amino acid, a very excellent whitening effect and anti-aging effect can be obtained. A skin external preparation that exerts its effects can be provided. In the external preparation for skin according to the present invention, glutathiones, which are active ingredients, can reach the melanocytes of the epidermis, dermis and basal layer of the epidermis, and exhibit excellent anti-aging and whitening effects at low concentrations. Therefore, there is no possibility that skin irritation or sensitization is exhibited.
【図１】本発明で用いる酸化型及び還元型のグルタチオ
ンのアルキルエステルとアミノ酸との塩類と、グルタチ
オン類のメラニン色素白色化作用を示す図である。FIG. 1 is a diagram showing salts of an oxidized and reduced glutathione alkyl ester and an amino acid used in the present invention, and the melanin pigment whitening action of glutathiones.
【図２】本発明で用いる酸化型及び還元型のグルタチオ
ンのアルキルエステルとアミノ酸との塩類、グルタチオ
ン類の活性酸素種による細胞傷害防御作用を示す図であ
る。FIG. 2 is a graph showing the cytotoxicity-protective effect of oxidized and reduced glutathione alkyl esters and amino acids used in the present invention, and glutathiones by reactive oxygen species.
１．作用例１ ２．作用例２ ３．作用例３ ４．作用例４ ５．作用例５ ６．作用例６ ７．作用例７ ８．比較例１ ９．比較例２ 1. Operation example 1 2. Example 2 of operation 3. Operation example 3 4. Operation example 4 5. Example of operation 5 6. Operation example 6 7. Operation example 7 8. Comparative Example 1 9. Comparative Example 2
フロントページの続き (51)Int.Cl.6 識別記号 庁内整理番号 ＦＩ 技術表示箇所 Ａ６１Ｋ 31/195 ＡＥＤ Ａ６１Ｋ 31/195 ＡＥＤ ＡＧＡ ＡＧＡ 31/215 ＡＤＳ 31/215 ＡＤＳ 31/70 ＡＧＺ 31/70 ＡＧＺ 38/00 ＡＤＡ 37/02 ＡＤＡ Continued on the front page (51) Int.Cl. 6 Identification number Agency reference number FI Technical display A61K 31/195 AED A61K 31/195 AED AGA AGA 31/215 ADS 31/215 ADS 31/70 AGZ 31/70 AGZ 38/00 ADA 37/02 ADA
Claims (6)
ルエステルとアミノ酸との塩類から選ばれる化合物の１
種又は２種以上を配合してなる皮膚外用剤。1. A compound selected from salts of oxidized and reduced glutathione alkyl esters and amino acids.
An external preparation for skin comprising a mixture of two or more species.
アミノ酸との塩類から選ばれる化合物の１種又は２種以
上と、還元型ニコチンアミドアデニンジヌクレオチド
（ＮＡＤＨ）及び／又は還元型ニコチンアミドアデニン
ジヌクレオチドリン酸（ＮＡＤＰＨ）とを併用してなる
皮膚外用剤。2. One or more compounds selected from salts of alkyl esters of oxidized glutathione and amino acids, reduced nicotinamide adenine dinucleotide (NADH) and / or reduced nicotinamide adenine dinucleotide An external preparation for skin which is used in combination with an acid (NADPH).
ピログルタミン酸，グリシン，アラニン，アスパラギン
酸，グルタミン酸から選ばれた１種又は２種以上である
ことを特徴とする請求項１又は請求項２に記載の皮膚外
用剤。(3) the amino acid is taurine, γ-aminobutyric acid,
The external preparation for skin according to claim 1 or 2, wherein the external preparation is one or more selected from pyroglutamic acid, glycine, alanine, aspartic acid, and glutamic acid.
エステルのアルキル基若しくはアルケニル基の炭素数
が、１〜１２までの炭素原子を含むことを特徴とする請
求項１〜請求項３に記載の皮膚外用剤。4. The skin according to claim 1, wherein the alkyl or alkenyl group of the alkyl ester of oxidized or reduced glutathione has 1 to 12 carbon atoms. External preparation.
エステルのアルキル基若しくはアルケニル基の炭素数
が、１〜６までの炭素原子を含むことを特徴とする請求
項１〜請求項３に記載の皮膚外用剤。5. The skin according to claim 1, wherein the alkyl group or alkenyl group of the alkyl ester of oxidized or reduced glutathione has 1 to 6 carbon atoms. External preparation.
る請求項１〜請求項６に記載の皮膚外用剤。6. The external preparation for skin according to claim 1, wherein the external preparation for skin is a cosmetic.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP23970296A JP4002313B2 (en) | 1996-08-21 | 1996-08-21 | Topical skin preparation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP23970296A JP4002313B2 (en) | 1996-08-21 | 1996-08-21 | Topical skin preparation |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH1059839A true JPH1059839A (en) | 1998-03-03 |
| JP4002313B2 JP4002313B2 (en) | 2007-10-31 |
Family
ID=17048659
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP23970296A Expired - Fee Related JP4002313B2 (en) | 1996-08-21 | 1996-08-21 | Topical skin preparation |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP4002313B2 (en) |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002076490A1 (en) * | 2001-02-08 | 2002-10-03 | Novelos Therapeutics, Inc. | Compounds comprising disulfide-containing peptides and nitrogenous bases, and medical uses thereof |
| JP2003500472A (en) * | 1999-05-27 | 2003-01-07 | サントゥル ナショナル ドゥ ラ ルシェルシュ シャーンティフィク（セ エン エール エス） | New antioxidants and their preparation and use |
| WO2004010968A1 (en) * | 2002-07-31 | 2004-02-05 | Yu Ruey J | Non-amphoteric glutathione derivative compositions for topical application |
| US7169412B2 (en) | 1998-11-23 | 2007-01-30 | Novelos Therapeutics, Inc. | Methods for production of the oxidized glutathione composite with cis-diamminedichloroplatinum and pharmaceutical compositions based thereof regulating metabolism, proliferation, differentiation and apoptotic mechanisms for normal and transformed cells |
| JP2008024600A (en) * | 2006-07-18 | 2008-02-07 | Naris Cosmetics Co Ltd | Inhibitor for acrolein adduct formation, skin anti-aging external preparation and anti-aging food and beverage containing the same |
| JP2014227393A (en) * | 2013-05-24 | 2014-12-08 | 株式会社ノエビア | Skin-whitening agent |
| CN110638827A (en) * | 2019-10-17 | 2020-01-03 | 泓博元生命科技（深圳）有限公司 | Application of NADH and its salt in preparing skin pigment inhibitor |
| CN114557912A (en) * | 2022-03-08 | 2022-05-31 | 广州市万千粉丝化妆品有限公司 | Application of nicotinamide adenine dinucleotide reduced coenzyme I (NADH) in anti-aging cosmetics |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101724604B1 (en) * | 2015-12-11 | 2017-04-07 | 재단법인 아산사회복지재단 | Whitening cosmetic composition to the skin containing GSH-MEE(Reduced glutathione monoethyl ester) |
-
1996
- 1996-08-21 JP JP23970296A patent/JP4002313B2/en not_active Expired - Fee Related
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7169412B2 (en) | 1998-11-23 | 2007-01-30 | Novelos Therapeutics, Inc. | Methods for production of the oxidized glutathione composite with cis-diamminedichloroplatinum and pharmaceutical compositions based thereof regulating metabolism, proliferation, differentiation and apoptotic mechanisms for normal and transformed cells |
| JP2003500472A (en) * | 1999-05-27 | 2003-01-07 | サントゥル ナショナル ドゥ ラ ルシェルシュ シャーンティフィク（セ エン エール エス） | New antioxidants and their preparation and use |
| WO2002076490A1 (en) * | 2001-02-08 | 2002-10-03 | Novelos Therapeutics, Inc. | Compounds comprising disulfide-containing peptides and nitrogenous bases, and medical uses thereof |
| WO2004010968A1 (en) * | 2002-07-31 | 2004-02-05 | Yu Ruey J | Non-amphoteric glutathione derivative compositions for topical application |
| JP2008024600A (en) * | 2006-07-18 | 2008-02-07 | Naris Cosmetics Co Ltd | Inhibitor for acrolein adduct formation, skin anti-aging external preparation and anti-aging food and beverage containing the same |
| JP2014227393A (en) * | 2013-05-24 | 2014-12-08 | 株式会社ノエビア | Skin-whitening agent |
| CN110638827A (en) * | 2019-10-17 | 2020-01-03 | 泓博元生命科技（深圳）有限公司 | Application of NADH and its salt in preparing skin pigment inhibitor |
| CN114557912A (en) * | 2022-03-08 | 2022-05-31 | 广州市万千粉丝化妆品有限公司 | Application of nicotinamide adenine dinucleotide reduced coenzyme I (NADH) in anti-aging cosmetics |
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| JP4002313B2 (en) | 2007-10-31 |
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