JP4393064B2 - How to use tyramine as a cosmetic ingredient to make white skin - Google Patents

How to use tyramine as a cosmetic ingredient to make white skin Download PDF

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JP4393064B2
JP4393064B2 JP2002520780A JP2002520780A JP4393064B2 JP 4393064 B2 JP4393064 B2 JP 4393064B2 JP 2002520780 A JP2002520780 A JP 2002520780A JP 2002520780 A JP2002520780 A JP 2002520780A JP 4393064 B2 JP4393064 B2 JP 4393064B2
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tyramine
cosmetic composition
whitening cosmetic
composition according
whitening
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リントナー カール
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セダーマ エス アー
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    • DTEXTILES; PAPER
    • D06TREATMENT OF TEXTILES OR THE LIKE; LAUNDERING; FLEXIBLE MATERIALS NOT OTHERWISE PROVIDED FOR
    • D06MTREATMENT, NOT PROVIDED FOR ELSEWHERE IN CLASS D06, OF FIBRES, THREADS, YARNS, FABRICS, FEATHERS OR FIBROUS GOODS MADE FROM SUCH MATERIALS
    • D06M23/00Treatment of fibres, threads, yarns, fabrics or fibrous goods made from such materials, characterised by the process
    • D06M23/12Processes in which the treating agent is incorporated in microcapsules
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
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    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/40Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing nitrogen
    • A61K8/41Amines
    • A61K8/415Aminophenols
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783Angiosperms [Magnoliophyta]
    • A61K8/9789Magnoliopsida [dicotyledons]
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    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
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    • A61K8/9783Angiosperms [Magnoliophyta]
    • A61K8/9794Liliopsida [monocotyledons]
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00Drugs for dermatological disorders
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00Drugs for dermatological disorders
    • A61P17/16Emollients or protectives, e.g. against radiation
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00Preparations for care of the skin
    • A61Q19/02Preparations for care of the skin for chemically bleaching or whitening the skin
    • DTEXTILES; PAPER
    • D06TREATMENT OF TEXTILES OR THE LIKE; LAUNDERING; FLEXIBLE MATERIALS NOT OTHERWISE PROVIDED FOR
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    • D06M13/00Treating fibres, threads, yarns, fabrics or fibrous goods made from such materials, with non-macromolecular organic compounds; Such treatment combined with mechanical treatment
    • D06M13/322Treating fibres, threads, yarns, fabrics or fibrous goods made from such materials, with non-macromolecular organic compounds; Such treatment combined with mechanical treatment with compounds containing nitrogen
    • D06M13/368Hydroxyalkylamines; Derivatives thereof, e.g. Kritchevsky bases
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
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Description

【0001】
表皮の特殊細胞で合成されるメラニンは色素細胞でこれによって皮膚の色が与えられる。普通の肌では、メラニンの合成は種々のファクターによって調整されている。このファクターの中でも良く知られているのが太陽光線で、メラニンは照射される紫外線(UV)(通常の生活での日光露出や自然の太陽光線照射、又はビューティーサロンのUV照射等)の線量に比例して増加する。これは日焼けとして現われる。比較的軽い或いは生理上の障害としてのメラニンが発生すると、そばかす、ほくろ、広がったしみ(クロアスマ、妊娠シミ)として現われる。老化による自然な生理的障害は多数あるが、そのうちメラニン色素の発生としては不規則に発現する老化性シミがある。
【0002】
生命に直接関わるのでない種々の不都合を調整するため、化粧品産業が独自の解決法を提供するのは、その当然の役割を果たしているといえる。
肌の色素過剰を手当てするため、多種の化学分子や物質が提供されてきた。しかし、そのうちで化粧品への適用として採用されたものは非常に少ない(かぶれ、法規上の規制、有毒性の問題などがあるため。)。そして、この選択に残ったものはあまり効果的でないのが一般的である。
化粧品又は皮膚薬品の成分としてチラミンを使用することが、この問題に有効な解答を与えることが明らかにされたので、本発明を特許として出願することにした。
【0003】
主として食物の中に存在する(生チーズ、キャベツ、じゃがいも、鰯、グルイエールチーズ、キアンティ、カマンベールチーズ等)チラミンは、今日まで製薬や化粧品には殆ど使用されていない。チラミンは調剤や診断において使用される(EP0795610)、或いは鬱症の治療(US4868218)、及び/又はアルツハイマー病やパーキンソン病(EP0406488)の治療、又はコレステロール生成防止(US4857522)剤として使用されてきた。
【0004】
生化学上の説明では、チラミンはドーパミンの直接前駆体である。ドーパに近い化学構造であるから、チラミンは抗チロシナーゼ効果を有するのではないか、そしてこれによって上記状況でおこるメラニン色素の生成を減少させるような製品が可能であろうという仮定を立てた。
【0005】
ところで、メラニン細胞は、チロシンがメラニンに変化するときの触媒酵素であるチロシナーゼとO−ジフェノールオキシダーゼを含んでいる。その反応は、以下の式で表される。
チロシン⇒ドーパ⇒ドーパキノン⇒ドーパクロム⇒インドールキノン⇒メラニン
【0006】
アルビノ(色素欠乏症)のヒトは、先天的にO−ジフェノールオキシダーゼが欠乏しており、これによってメラニン形成ができず、従って皮膚色素が欠如しているわけである。
【0007】
本発明の目的は、チラミンがそれ自身はドーパに変化することなく、システムを阻害することによって、メラニン形成を減少するように作用するということを証明するものである。すなわち、メラニンは代替物質ではなく競争阻害物質である。
【0008】
チラミンは様々な素材料から得ることができる。チラミンを含有する植物は全て原料として利用することができるが、その中でも特に、マンダリン(Citrus reticulata)はチラミンを豊富に含んでいる(Wheaton TA & Stewart I(1970) Lloydia 33:244-254)ので、植物体全体または一部から抽出によってチラミンを得ることができる。
【0009】
植物全体又はその一部(15g)を85gの蒸留水に浸漬し、常温で48時間スクリュー攪拌する。植物は分散して水の体積と同量になる。濾過(800μm)後、抽出液はそのまま使用することができる。
或いは、後述する形態に利用できるように、従来の物質乾燥方法によって乾燥抽出体にすることもできる。
【0010】
高速液体クロマトグラフィーによる分析によって、このようにして得られた抽出体はチラミンを含んでいることが示される。植物の原産地や収穫時期によってチラミン含有量が異なることがあるが、この分野の専門家であればチラミン含有量が一定になるように抽出体を調整することができる。
【0011】
マンダリン(Citrus reticulata)抽出からチラミンを収集する方法は、上述のものに限定されない。例えば、浸漬、煎じ法、浸出、還流、超音波やマイクロ波による抽出、洗浄法等の方法でエキス抽出が可能である。方法は例示したものに限定されない。
また抽出溶剤としては、上記のものに限定されず、水、プロピレングリコール、ブチレングリコール、グリセリン、ポリエチレングリコール、メチルエーテルジグリコール、エチルエーテルジグリコール、環状ポリアルコール、エトキシルジグリコール、プロポキシルジグリコール、アルコール(メタノール、エタノール、プロパノール、ブタノール)、またはこれらの混合溶媒を使用することができる。
【0012】
本発明において期待される活性を得るためのチラミンは、以下のように要約される。
・植物の抽出エキスから得られ、例えば陳皮(Citrus unshiu Mare.)、好ましくはマンダリン(Citrus reticulata)から抽出されるものが好ましい。
・あらゆる方法の化学合成からも得られる。
・単独または他の活性分子と組み合わせて使用することができる。
・精製され、リポゾーム等のサポート媒体に付けられる。
・1乃至24個の炭素原子を含む、直鎖または分岐の、飽和化合物又は不飽和化合物の、或いはヒドロキシル基化合物又はイオウ化合物を含むか含まない、アルキル基分子鎖によって、OH基又はNH基に接合したもの。
【0013】
チラミンは化粧品や皮膚用薬品に使用される植物成分の含有形態として配合される。例えば、以下に二つの成分構成を例示する。
【0014】
例1 日常クリーム
ボルポS20 2.4g
ボルポS2 2.6g
プロステアリル15 8.0g
ミツロウ 0.5g
アビル(登録商標)ZP2434 3.0g
プロピレングリコール 3.0g
カーボポール(登録商標)1941 0.25g
トリエタノールアミン 0.25g
チラミン 5.0g
水分及び防腐剤を加えて100gとする。
【0015】
例2 ボディーローション美白デオドラント
クリレット3 2.5g
ノボル 0.9g
フルイラン 2.5g
カーボポール940 0.3g
ミツロウ 2.0g
トリエタノールアミン 0.1g
グリセリン 5.0g
チラミン 3.0g
水分及び防腐剤を加えて100gとする。
【0016】
以下の例において、チラミンの美白作用効果を証明する。
例3 ヒトチロシナーゼ阻害作用(インビトロ試験)
この例ではドーパミンオキシダーゼの活性が明らかにされる。培養されたヒト正常メラニン細胞(MNH)をイゲパル緩衝液と超音波で溶解させ、この抽出細胞をそれぞれ重量濃度0.1%、0.3%、1%のチラミンを含む緩衝液およびチラミンを含まないコントロール液に接触させた。
チロシナーゼの活性度は、L−ドーパオキシダーゼ活性から、L−ドーパからつくられたドーパクロムの量を波長490nmにおける光学スペクトル濃度測定によって定量化した。反応はL−ドーパを加えることにより開始された。
波長490nmで、開始より1時間後までは10分ごとに、2時間後までは30分ごとに動的変化を追跡した。異なる緩衝液における8回のテスト結果をコントロール液と比較した。阻害作用の平均結果をパーセント表示で以下の表1に記載する。
【0017】
【表1】

Figure 0004393064
【0018】
この表1の結果によれば、チラミンの存在でチロシナーゼ活性が非常に低下することが示されている。チラミンの存在、非存在がこの実験の唯一の変数であるから、観察された効果はチラミンに由来するものであることは明らかである。また観察されたこのシステムの効果は、明らかに濃度依存性を示しているから、チラミンに特徴付けられる効果といえる。
またこの実験はL−ドーパおよびチラミンなしのテストを行い、システムの安定性が確認されている。
【0019】
例4 チロシナーゼの阻害作用(インビトロ実験)
生理的効果を立証するため、ヒト細胞を使用することは理にかなっているが、試料提供者の個人差があるので、時として結果が対象ごとに大差を示すことは周知の事実である。
この理由から、前記のテスト結果は、チラミンのチロシナーゼに対する阻害効果を疑いなく示すものであるが、他のモデルを使用して別の実験を行った。
この例で用いた細胞は、メラニンを多数生成し、チロシナーゼ活性やメラニン合成の活性に関する研究に広く使用されているメラノーマB16細胞である。
この細胞を培養プレートのくぼみ(1プレートにつき24個のくぼみが設けてある)に植えつけ、37℃で24時間培養した。洗浄後、細胞はチラミン濃度がそれぞれ0.01%、0.03%の緩衝液、またチラミンを含まないコントロール液に48時間接触させた。
比較例として、濃度0.01×10−2%のハイドロキノン液を用いた。前記実験と同様、細胞はイゲパル緩衝液で超音波を用い溶解して測定した。活性チロシナーゼ量は共存するタンパク質量当りに換算することで標準化した。
【0020】
【表2】
Figure 0004393064
【0021】
この結果は、チラミン存在のためチロシナーゼ活性が非常に減少していること、それは相当する濃度のハイドロキノンによるものより良い効果を示している。チラミンの存在、非存在がこの実験の唯一の変数であるから、観察された効果はチラミン由来のものであることは明らかである。また実験は二種類の異なる濃度のチラミンで行われたに過ぎないが、観察されたこのシステムの効果は明らかに濃度依存性を示し、チラミンに特徴付けられる効果といえる。
【0022】
例5.合成チラミンの阻害作用(インビトロ試験)
培養(60%と80%の混合)されたヒト正常メラニン細胞(MNH)を、それぞれチラミン濃度が0.003%と0.01%の緩衝液、およびチラミンを含まないコントロール液中で接触させた。8日間緩衝液と接触させ、この期間中、細胞を維持するため毎日同じ濃度(重量濃度で0.003%と0.01%)の緩衝液を更新した。
この期間中、半数のプレートを五回にわたって紫外線UV−B(20mJ/cm)で照射した。他の半数のプレートは照射しなかった。
8日後、細胞を培養から取り出し洗浄した。抽出後、細胞間メラニンの量を従来の光学スペクトル濃度測定法で測定した。この測定値は、一定細胞数当りに換算して標準化した。
【0023】
以下に、実験後の平均阻害効果率パーセンテージ(5回の異なるテストの)結果を、同じ時間で実験した比較例と比較して示す。
【表3】
Figure 0004393064
【0024】
この結果は、
・一方、UV−B照射がない場合、チラミン量に基づいたメラニンの減少効果が示されている。すなわち形成されるメラニンの全体量が減少されるから期待される美白効果となる。
・他方、UV−Bを五回照射した後も、チラミンの存在でメラニン生成の阻害効果は維持され、比較例より低い値を示している。このように、紫外線照射による新しいメラニンの合成促進効果(比較例でほぼ2倍)は限定される。
【0025】
チラミンの存在、非存在がこの実験の唯一の変数であるから、観察された効果はチラミン由来のものであることは明らかである。また実験は二つの違う濃度のチラミンでしか行われていないが、観察されたこのシステムの効果は、明らかに濃度依存性を示しているから、チラミンに特徴付けられる効果であるといえる。
【0026】
例6.抗シミ作用(臨床試験)
この実験は、年齢が45歳から65歳で、肌や手に老化性シミのある女性志願者を対象に行った。
被験者の一方の手の五ケ所のシミを選び、色と大きさを正確に測定した後、例1に記述したテスト物質(5%)含有クリームを1.5ヶ月間にわたって一方の手(どちらでもよい)にだけ塗り、他方の手は比較例として、テスト物質を含まないクリームを塗った。結果を標準化するため、当然、シミの無い皮膚5箇所の色素測定も行った。1ヶ月後、および1.5ヶ月後に皮膚の色素定量化を行った。メラニン色素の量はメキサ測定器で測定した。この機器はセンサー中に内蔵された異なる三色光を発する特殊ランプでメラニン量を測るものである。
テスト物質を塗った手の老化性シミの色は、チラミンを含まないクリームで処理した手に比較して、一ヶ月で6.4%、1.5ヶ月後で24%低くなった。
この実験は2000年春、特に太陽光の多い時期に行われたことを考慮すると、結果の著しい効果を示しているといえる。
【0027】
これらの例はいずれも美白効果および自然光または人工のUV照射による皮膚の着色を弱める効果をはっきりと証明している。
最終の化粧品または皮膚用薬品成分として、チラミンの重量濃度は0.001〜10%の範囲で、好ましくは0.01〜7.0%がよい。
チラミンは化粧品や皮膚用薬品で使用されるあらゆる形態で使用することができる。すなわち、O/W又はW/Oエマルション、乳液、ローション、ポマード、ヘアローション、シャンプー、石鹸、スティック、ペンシル、スプレー、ボディオイルなどの形態で、使用することができる。但し、これらの形態に限定されるものではない。
【0028】
チラミンを化粧品用のキャリアに取り込むことも可能である。例えば、リポゾーム、マイクロエマルション、マクロ粒子、ミクロ粒子、ナノ粒子、あるいはマクロカプセル、ミクロカプセル、ナノカプセルなどの媒体に取り込んだり、粉状有機ポリマーやタルク、ベントナイト等の鉱物サポートに吸収させたりすることもできる。
チラミンは通常化粧品で使用されるその他のいかなる材料とも組み合わせて化粧品成分とすることができる。例えば、天然抽出または合成の脂質、ゲル状および粘質のポリマー、界面活性剤およびエマルション、水溶性活性剤または脂溶性活性剤、他の植物からの抽出物、繊維抽出物、海産物抽出物などを例示することができる。
【0029】
チラミンを含む化粧品または皮膚用薬品としてはクリーム、芳香剤、ゲル、ローション状、またはクリーム状の日焼け止めや日焼け止め処置薬、アフターシェービング、脱毛剤、脱毛後手入れクリームなどがある。
チラミンは、肌の手入れ、特に肌を色白にしたり、人工または自然の紫外線照射時の色素沈着を軽減するあらゆる化粧品または皮膚用薬品の中に混入したりすることができる。
化粧品または皮膚用薬品のこれらの成分は、肌の手入れ、特に肌を白くしたり、人工または自然の紫外線照射時の色素沈着を軽減したりするための薬品下地に使用される。
【0030】
チラミンは化粧品または皮膚用薬品の中に単体または複合体として使用され、マクロ、ミクロ、ナノ粒子やマクロ、ミクロ、ナノカプセルのかたちで化学結合、混入、吸収、吸着させ、織物、天然または合成の繊維、羊毛など昼夜を問わず肌や毛髪と直接接触することによって局所的にチラミンを連続放出するような着物や下着を仕立てるものに用いるあらゆる材料の中に使用される。[0001]
Melanin synthesized in special cells of the epidermis is a pigment cell, which gives the skin color. In normal skin, melanin synthesis is regulated by various factors. Of these factors, sunlight is well known, and melanin is the dose of ultraviolet rays (UV) that are irradiated (sunlight exposure in normal life, natural sunlight irradiation, or beauty salon UV irradiation, etc.). Increase proportionally. This appears as a tan. When melanin occurs as a relatively mild or physiological disorder, it appears as freckles, moles, spread stains (cloasma, pregnancy spots). There are many natural physiological disorders due to aging. Among them, there are aging spots that appear irregularly as melanin development.
[0002]
It can be said that the cosmetic industry plays its own role in order to adjust various inconveniences not directly related to life.
A variety of chemical molecules and substances have been provided to treat excess skin pigment. However, very few of them have been adopted for cosmetic applications (because of rashes, regulatory restrictions, toxicity issues, etc.). And what remains of this choice is generally not very effective.
Since the use of tyramine as a cosmetic or dermatological ingredient has been shown to provide an effective answer to this problem, the present invention was filed as a patent.
[0003]
Tyramine, which is mainly present in food (raw cheese, cabbage, potatoes, potatoes, gruyere cheese, chianti, camembert cheese, etc.) is rarely used in pharmaceuticals and cosmetics to date. Tyramine has been used in formulation and diagnosis (EP 07956610), or treatment of depression (US 4868218), and / or treatment of Alzheimer's disease or Parkinson's disease (EP 0406488), or cholesterol production prevention (US 4857522).
[0004]
In the biochemical description, tyramine is a direct precursor of dopamine. Since it has a chemical structure close to that of dopa, it was hypothesized that tyramine would have an anti-tyrosinase effect and that this would enable a product that would reduce the production of melanin pigments occurring in the above situation.
[0005]
By the way, melanocytes contain tyrosinase and O-diphenol oxidase which are catalytic enzymes when tyrosine changes to melanin. The reaction is represented by the following formula.
Tyrosine ⇒ Dopa ⇒ Dopaquinone ⇒ Dopachrome ⇒ Indolequinone ⇒ Melanin [0006]
Humans with albino (pigment deficiency) are congenitally deficient in O-diphenol oxidase, thereby failing to form melanin and thus lacking skin pigment.
[0007]
The purpose of the present invention is to prove that tyramine acts to reduce melanogenesis by inhibiting the system without itself changing to dopa. That is, melanin is not a substitute but a competitive inhibitor.
[0008]
Tyramine can be obtained from a variety of raw materials. All plants containing tyramine can be used as raw materials, but mandarin (Citrus reticulata) is particularly rich in tyramine (Wheaton TA & Stewart I (1970) Lloydia 33: 244-254). Tyramine can be obtained by extraction from all or part of the plant body.
[0009]
The whole plant or a part thereof (15 g) is immersed in 85 g of distilled water, and is stirred with a screw at room temperature for 48 hours. Plants disperse to the same volume as water. After filtration (800 μm), the extract can be used as it is.
Or it can also be made into a dry extract by the conventional substance drying method so that it can utilize for the form mentioned later.
[0010]
Analysis by high performance liquid chromatography shows that the extract thus obtained contains tyramine. The tyramine content may vary depending on the plant's origin and harvest time, but an expert in this field can adjust the extract so that the tyramine content is constant.
[0011]
The method of collecting tyramine from mandarin (Citrus reticulata) extraction is not limited to the one described above. For example, the extract can be extracted by a method such as dipping, decoction, leaching, reflux, extraction with ultrasonic waves or microwaves, or a washing method. The method is not limited to that illustrated.
Further, the extraction solvent is not limited to the above, but water, propylene glycol, butylene glycol, glycerin, polyethylene glycol, methyl ether diglycol, ethyl ether diglycol, cyclic polyalcohol, ethoxyl diglycol, propoxyl diglycol, alcohol (Methanol, ethanol, propanol, butanol) or a mixed solvent thereof can be used.
[0012]
Tyramine for obtaining the activity expected in the present invention is summarized as follows.
-It is obtained from an extract of a plant, for example, one extracted from citrus unshiu Mare., Preferably mandarin (Citrus reticulata).
• Can be obtained from any method of chemical synthesis.
Can be used alone or in combination with other active molecules.
・ Purified and attached to a support medium such as liposome.
· Containing 1 to 24 carbon atoms, linear or branched, saturated or unsaturated compounds, or with or without hydroxyl groups or sulfur compounds, depending on the alkyl group molecular chain, OH groups or NH 2 groups What was joined to.
[0013]
Tyramine is blended as a plant component used in cosmetics and dermatological agents. For example, two component structures are illustrated below.
[0014]
Example 1 Daily cream Volpo S20 2.4g
Volpo S2 2.6g
Prostearyl 15 8.0g
Beeswax 0.5g
Avil (registered trademark) ZP2434 3.0 g
Propylene glycol 3.0g
Carbopol (registered trademark) 1941 0.25g
0.25 g of triethanolamine
Tyramine 5.0g
Add water and preservatives to 100 g.
[0015]
Example 2 Body Lotion Whitening Deodorant Krillet 3 2.5g
Noboru 0.9g
Fluilan 2.5g
Carbopol 940 0.3g
Beeswax 2.0g
Triethanolamine 0.1g
Glycerin 5.0g
Tyramine 3.0g
Add water and preservatives to 100 g.
[0016]
The following example demonstrates the whitening effect of tyramine.
Example 3 Human tyrosinase inhibitory action (in vitro test)
In this example, the activity of dopamine oxidase is revealed. Cultured human normal melanocytes (MNH) are lysed with Igepal buffer and ultrasound, and the extracted cells contain buffer solution containing tyramine at a weight concentration of 0.1%, 0.3% and 1% and tyramine, respectively. No contact with control solution.
The activity of tyrosinase was quantified from the L-dopa oxidase activity by measuring the amount of dopachrome produced from L-dopa by optical spectral densitometry at a wavelength of 490 nm. The reaction was started by adding L-dopa.
Dynamic changes were followed at a wavelength of 490 nm every 10 minutes up to 1 hour after the start and every 30 minutes up to 2 hours. Eight test results in different buffers were compared with the control solution. The average results of the inhibitory action are listed in Table 1 below as a percentage.
[0017]
[Table 1]
Figure 0004393064
[0018]
The results in Table 1 indicate that tyrosinase activity is greatly reduced in the presence of tyramine. Since the presence or absence of tyramine is the only variable in this experiment, it is clear that the observed effect is derived from tyramine. In addition, the observed effect of this system clearly shows concentration dependence, and can be said to be an effect characterized by tyramine.
The experiment was also tested without L-dopa and tyramine, confirming the stability of the system.
[0019]
Example 4 Inhibitory action of tyrosinase (in vitro experiment)
Although it makes sense to use human cells to demonstrate physiological effects, it is a well-known fact that the results sometimes vary greatly from subject to subject due to individual differences among sample providers.
For this reason, the test results clearly show the inhibitory effect of tyramine on tyrosinase, but other experiments were performed using other models.
The cells used in this example are melanoma B16 cells that produce many melanins and are widely used for research on tyrosinase activity and melanin synthesis activity.
The cells were inoculated into the wells of the culture plate (24 wells were provided per plate) and cultured at 37 ° C. for 24 hours. After washing, the cells were contacted with a buffer solution having a tyramine concentration of 0.01% and 0.03%, respectively, and a control solution not containing tyramine for 48 hours.
As a comparative example, a hydroquinone liquid having a concentration of 0.01 × 10 −2 % was used. As in the previous experiment, cells were measured by lysis using ultrasound with Igepal buffer. The amount of active tyrosinase was standardized by converting to the amount of coexisting protein.
[0020]
[Table 2]
Figure 0004393064
[0021]
This result shows that tyrosinase activity is greatly reduced due to the presence of tyramine, which is a better effect than with the corresponding concentration of hydroquinone. Since the presence or absence of tyramine is the only variable in this experiment, it is clear that the observed effect is from tyramine. The experiment was only conducted with two different concentrations of tyramine, but the observed effects of this system are clearly concentration-dependent and can be characterized as tyramine.
[0022]
Example 5. Inhibitory action of synthetic tyramine (in vitro test)
Cultured (60% and 80% mixed) human normal melanocytes (MNH) were contacted in buffers with tyramine concentrations of 0.003% and 0.01%, respectively, and a control solution without tyramine. . The buffer was renewed at the same concentration (0.003% and 0.01% by weight) daily to maintain the cells during this period in contact with the buffer for 8 days.
During this period, half of the plates were irradiated five times with UV-B (20 mJ / cm 2 ). The other half of the plates were not irradiated.
After 8 days, the cells were removed from the culture and washed. After extraction, the amount of intercellular melanin was measured by a conventional optical spectral densitometry method. This measured value was standardized by converting per fixed number of cells.
[0023]
Below, the mean percentage inhibition rate after experiment (from 5 different tests) results are shown in comparison with the comparative examples run at the same time.
[Table 3]
Figure 0004393064
[0024]
The result is
-On the other hand, when there is no UV-B irradiation, the reduction effect of melanin based on the amount of tyramine is shown. That is, the whitening effect is expected because the total amount of melanin formed is reduced.
-On the other hand, even after UV-B was irradiated five times, the inhibitory effect of melanin production was maintained in the presence of tyramine, indicating a lower value than in the comparative example. Thus, the effect of promoting the synthesis of new melanin by ultraviolet irradiation (approximately twice in the comparative example) is limited.
[0025]
Since the presence or absence of tyramine is the only variable in this experiment, it is clear that the observed effect is from tyramine. The experiment was only conducted at two different concentrations of tyramine, but the observed effects of this system are clearly concentration-dependent and can be said to be characterized by tyramine.
[0026]
Example 6 Anti-staining effect (clinical trial)
This experiment was conducted on female volunteers aged 45 to 65 who have aging spots on their skin and hands.
After five spots on one hand of the subject were selected and the color and size were accurately measured, the cream containing the test substance (5%) described in Example 1 was applied to one hand (either The other hand was applied with a cream containing no test substance as a comparative example. In order to standardize the results, naturally, pigment measurement was also performed on five spots without skin spots. Skin pigment quantification was performed after 1 month and 1.5 months. The amount of melanin pigment was measured with a mexa meter. This device measures the amount of melanin with a special lamp that emits three different colors of light built into the sensor.
The color of the aging spots on the hands coated with the test substance was 6.4% lower in one month and 24% lower after 1.5 months compared to hands treated with cream without tyramine.
Considering that this experiment was carried out in the spring of 2000, especially during the day when there was a lot of sunlight, it can be said that the result shows a remarkable effect.
[0027]
Both of these examples clearly demonstrate a whitening effect and an effect that attenuates skin coloration by natural light or artificial UV irradiation.
As a final cosmetic or dermatological chemical component, the weight concentration of tyramine is in the range of 0.001 to 10%, preferably 0.01 to 7.0%.
Tyramine can be used in any form used in cosmetics and dermatological agents. That is, it can be used in the form of O / W or W / O emulsion, emulsion, lotion, pomade, hair lotion, shampoo, soap, stick, pencil, spray, body oil and the like. However, it is not limited to these forms.
[0028]
It is also possible to incorporate tyramine into a cosmetic carrier. For example, it can be taken into a liposome, microemulsion, macroparticle, microparticle, nanoparticle, or a medium such as macrocapsule, microcapsule, nanocapsule, or absorbed into a mineral support such as powdered organic polymer, talc, or bentonite. You can also.
Tyramine can be combined with any other material normally used in cosmetics to make a cosmetic ingredient. For example, natural or synthetic lipids, gel-like and viscous polymers, surfactants and emulsions, water-soluble or fat-soluble active agents, extracts from other plants, fiber extracts, marine product extracts, etc. It can be illustrated.
[0029]
Cosmetic or dermatological agents containing tyramine include creams, fragrances, gels, lotions, or creamy sunscreens and sunscreens, aftershaves, hair removers, post-hair removal care creams, and the like.
Tyramine can be incorporated into any cosmetic or dermatological agent that cares for the skin, in particular the skin becomes pale, or reduces pigmentation during artificial or natural UV irradiation.
These components of cosmetic or dermatological chemicals are used in drug bases for skin care, in particular for whitening the skin and reducing pigmentation during artificial or natural UV irradiation.
[0030]
Tyramine is used as a simple substance or complex in cosmetics or dermatological chemicals, chemically bonded, mixed, absorbed, adsorbed in the form of macro, micro, nano particles or macro, micro, nano capsules, textile, natural or synthetic It is used in all materials used for making kimonos and underwear that continuously release tyramine locally by direct contact with skin and hair, such as fiber and wool.

Claims (8)

陳皮(Citrus unshiu Mare.)またはマンダリン(Citrus reticulata)から抽出されるチラミンをヒトチロシナーゼ阻害活性成分として、0.01〜7.0重量%含有する美白化粧料組成物。 A whitening cosmetic composition containing 0.01 to 7.0% by weight of tyramine extracted from citrus unshiu Mare. Or mandarin (Citrus reticulata) as a human tyrosinase inhibitory active ingredient . 合成又は発酵工程で生成されるチラミンをヒトチロシナーゼ阻害活性成分として、0.01〜7.0重量%含有する美白化粧料組成物。 A whitening cosmetic composition containing 0.01 to 7.0% by weight of tyramine produced in a synthesis or fermentation process as a human tyrosinase inhibitory active ingredient . 形態が、O/Wエマルション、W/Oエマルション、乳液、ローション、ポマード、ヘアローション、シャンプー、石鹸、スティック、ペンシル、スプレー、ボディオイルから選択されるいずれか一つである請求項1又は2のいずれかに記載の美白化粧料組成物。 The form is any one selected from O / W emulsion, W / O emulsion, emulsion, lotion, pomade, hair lotion, shampoo, soap, stick, pencil, spray, and body oil. The whitening cosmetic composition according to any one of the above . チラミンが、リポソーム、マイクロエマルジョン、マクロ粒子、ミクロ粒子、ナノ粒子、マクロカプセル、ミクロカプセル、ナノカプセルから選択されるいずれか一つの化粧品用キャリアの中に取り込まれたものであることを特徴とする請求項1乃至3いずれか記載の美白化粧料組成物。 Tyramine is incorporated in any one cosmetic carrier selected from liposomes, microemulsions, macroparticles, microparticles, nanoparticles, macrocapsules, microcapsules, nanocapsules The whitening cosmetic composition according to any one of claims 1 to 3 . チラミンが、粉体有機ポリマー、タルク、ベントナイトから選択されるいずれか一つに吸収されていることを特徴とする請求項1乃至3いずれか記載の美白化粧料組成物。 The whitening cosmetic composition according to any one of claims 1 to 3, wherein tyramine is absorbed in any one selected from powdered organic polymer, talc, and bentonite . チラミンが、天然抽出又は合成の脂質、ゲル状及び粘性のポリマー、界面活性剤及び乳化剤、水溶性活性剤又は脂溶性活性剤、組織抽出物、海産品抽出物から選択されるいずれか一つと共に使用されることを特徴とする請求項1乃至5のいずれかに記載の美白化粧料組成物。 Tyramine with any one selected from natural or synthetic lipids, gel and viscous polymers, surfactants and emulsifiers, water-soluble or fat-soluble active agents, tissue extracts, marine product extracts The whitening cosmetic composition according to any one of claims 1 to 5, wherein the whitening cosmetic composition is used. 化粧料が、クリーム、芳香剤、ジェル状又はローション状又はクリーム状の日焼け止めや日焼け処理薬、アフターシェービング、脱毛剤、又は脱毛後手入れクリームであることを特徴とする請求項1乃至6のいずれかに記載の美白化粧料組成物。 The cosmetic is a cream, a fragrance, a gel-like or lotion-like or cream-like sunscreen or sunscreen agent, aftershave, a hair removal agent, or a hair care cream after hair removal. A whitening cosmetic composition according to claim 1. 肌を色白又は薄い色にしたり、人工又は自然の紫外線に露出されたときの色素沈着を軽減することを特徴とする請求項1乃至7のいずれかに記載の美白化粧料組成物。 The whitening cosmetic composition according to any one of claims 1 to 7, wherein the skin whitening or pale color or pigmentation when exposed to artificial or natural ultraviolet rays is reduced .
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