JP6792270B2 - Whitening composition - Google Patents
Whitening composition Download PDFInfo
- Publication number
- JP6792270B2 JP6792270B2 JP2018002679A JP2018002679A JP6792270B2 JP 6792270 B2 JP6792270 B2 JP 6792270B2 JP 2018002679 A JP2018002679 A JP 2018002679A JP 2018002679 A JP2018002679 A JP 2018002679A JP 6792270 B2 JP6792270 B2 JP 6792270B2
- Authority
- JP
- Japan
- Prior art keywords
- lactic acid
- tyrosinase
- inhibitory activity
- extract
- tyrosinase inhibitory
- 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.)
- Active
Links
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Non-Alcoholic Beverages (AREA)
- Cosmetics (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Description
本発明は美白用組成物に関する。 The present invention relates to a whitening composition.
胎盤の抽出物（プラセンタ抽出物）は、種々の作用効果を有することから多くの化粧用組成物や食品組成物、医薬品等に用いられている。例えば、特許文献１（特開２００２−１７９５２３号公報）や特許文献２（特開２００２−１８７８１２号公報）にはブタやウマの胎盤抽出物がチロシナーゼ活性阻害作用や肌荒れ改善作用を有するので、化粧用組成物として好適に使用し得ることが記載されている。 The placenta extract (placenta extract) is used in many cosmetic compositions, food compositions, pharmaceuticals, etc. because it has various effects. For example, in Patent Document 1 (Japanese Patent Laid-Open No. 2002-179523) and Patent Document 2 (Japanese Patent Laid-Open No. 2002-187812), placenta extracts of pigs and horses have a tyrosinase activity inhibitory action and a rough skin improving action. It is described that it can be suitably used as a composition for use.
その一方で、例えば特許文献３（特開２０１６−４４１４１号公報）には、プラセンタ抽出物にはチロシナーゼ活性阻害作用がほとんどないか、あるいは極めて活性が弱いものがあるが、遊離チロシン含有量を所定量以下とすると、高いチロシナーゼ活性阻害作用が得られることが記載されている。 On the other hand, for example, in Patent Document 3 (Japanese Unexamined Patent Publication No. 2016-44141), the placenta extract has almost no inhibitory effect on tyrosinase activity or has extremely weak activity, but the content of free tyrosine is determined. It is described that a high tyrosinase activity inhibitory effect can be obtained when the amount is less than the fixed amount.
ところで、特許文献４（特開２００６−８５６６号公報）には、パイナップル果汁などのある種の果汁の乳酸菌発酵物にもチロシナーゼ活性阻害作用が認められることが記載されている。しかしながら、果実から果汁を搾り取った後には、果汁の絞り滓を始め、果皮やパイナップルの果芯部などの各種の廃棄物が得られるために、これらの廃棄物を有効に利用することが重要な課題となっている。 By the way, Patent Document 4 (Japanese Unexamined Patent Publication No. 2006-8566) describes that a lactic acid bacterium fermented product of a certain fruit juice such as pineapple juice also has an inhibitory effect on tyrosinase activity. However, after squeezing the fruit juice from the fruit, various wastes such as the squeezed residue of the fruit juice, the peel and the core of the pineapple can be obtained, so it is important to make effective use of these wastes. It has become an issue.
これらの廃棄物の利用法として、例えば、特許文献５（特開２０１６−０４４１２９号公報）には、パイナップルの果芯をセルラーゼ処理することで保湿作用のある果芯処理物が得られることが記載されている。 As a method of using these wastes, for example, Patent Document 5 (Japanese Unexamined Patent Publication No. 2016-044129) describes that a fruit core-treated product having a moisturizing effect can be obtained by cellulase-treating the fruit core of pineapple. Has been done.
しかしながら、パイナップル果芯のようなパイナップルの非可食部の乳酸菌培養物について、チロシナーゼ活性阻害作用などの機能性が見いだされたとの報告は見当たらない。 However, there are no reports that functionality such as tyrosinase activity inhibitory action was found in lactic acid bacteria cultures in the non-edible part of pineapple such as pineapple fruit core.
本願発明者らは、上記背景技術に鑑みて鋭意研究を行ったところ、パイナップルの果芯の乳酸菌による培養物にチロシナーゼに対する阻害活性を見いだした。 As a result of diligent research in view of the above background technology, the inventors of the present application found an inhibitory activity against tyrosinase in a culture of pineapple fruit core by lactic acid bacteria.
また、本願発明者らが試験したところ、遊離チロシン含有量が低いプラセンタ抽出物であっても、チロシナーゼ活性阻害作用がほとんどないか、あるいは極めて活性が弱いことが確認された。しかしながら、チロシナーゼ活性阻害作用がほとんどないか、あるいは極めて活性が弱いプラセンタ抽出物であっても、パイナップル果芯の乳酸菌培養物を組み合わせたところ、より強いチロシナーゼ活性阻害作用を見いだせた。 In addition, as a result of tests by the inventors of the present application, it was confirmed that even the placenta extract having a low free tyrosine content has almost no tyrosinase activity inhibitory action or extremely weak activity. However, even in the placenta extract having almost no tyrosinase activity inhibitory action or extremely weak activity, a stronger tyrosinase activity inhibitory action was found when the lactic acid bacterium culture of pineapple fruit core was combined.
すなわち、本願発明が解決しようとする課題はチロシナーゼ活性阻害作用に基づく新規な美白用組成物を提供することにある。 That is, the problem to be solved by the present invention is to provide a novel whitening composition based on the tyrosinase activity inhibitory action.
本願の主たる発明は、酵素処理、加熱処理又は加水分解処理のうち何れか１又は２以上の処理を経て得られたプラセンタ抽出物を用いて、組成物中に含まれるチロシナーゼ活性阻害剤のチロシナーゼ阻害活性よりも阻害活性が高められた美白用組成物を提供する。 The main invention of the present application is to inhibit tyrosinase of a tyrosinase activity inhibitor contained in a composition by using a placenta extract obtained through any one or more treatments of enzyme treatment, heat treatment or hydrolysis treatment. Provided is a whitening composition in which the inhibitory activity is enhanced rather than the activity.
本願の主たる発明によると、チロシナーゼ活性阻害作用がほとんどないと言えるプラセンタ抽出物であっても、他のチロシナーゼ活性阻害剤と組み合わせることで、より効果の強い美白用組成物が提供される。 According to the main invention of the present application, even a placenta extract which can be said to have almost no tyrosinase activity inhibitory action can be combined with another tyrosinase activity inhibitor to provide a more effective whitening composition.
本願発明に係る美白用組成物はチロシナーゼ阻害活性剤とプラセンタエキスを含む。用いられ得るチロシナーゼ阻害活性剤は、特に限定されることはなく、チロシナーゼ阻害活性を有する化合物、各種の動植物・微生物の抽出物やそれらの培養物、それらの加水分解物などであり得る。チロシナーゼ阻害活性を有する化合物としては、アルブチンやコウジ酸、グルタチオン、エラグ酸などが、動植物・微生物の抽出物としては、藤茶抽出物（特開２００２−３７０９６２号公報）やタイム・レモンバームの抽出物（特開平０６−１９９６４７号公報）が、動植物の発酵物としては緑茶抽出物の麹菌培養物（特開２０１２−９２１００号公報）、パイナップル果汁やブドウ果汁の乳酸菌培養物（特許文献２００６−８５６６号公報）が、また加水分解物としては、シロウリの抽出物の加水分解物（特開２０１１−２５６１４７号公報）が例示される。これら以外にもチロシナーゼ阻害作用があるとして市販されているライチ種子エキス、月見草エキス、キウイ種子エキス、温オウゴンエキス、イチョウ葉エキス、ラクトフェリンなども使用され得る。また、本願発明に係るチロシナーゼ活性阻害剤であるパイナップル果芯の乳酸菌培養物や、果芯を含むパイナップル果実の乳酸菌培養物でもあり得る。美白用組成物には、これらのチロシナーゼ活性阻害剤の１種又は２種以上が用いられる。 The whitening composition according to the present invention contains a tyrosinase inhibitory activator and a placenta extract. The tyrosinase inhibitory activator that can be used is not particularly limited, and may be a compound having a tyrosinase inhibitory activity, extracts of various animals, plants and microorganisms, cultures thereof, hydrolysates thereof and the like. Arbutin, kojiic acid, glutathione, ellagic acid and the like are used as compounds having tyrosinase inhibitory activity, and wisteria tea extract (Japanese Patent Laid-Open No. 2002-370962) and thyme lemon balm extract are used as extracts of animals, plants and microorganisms. (Japanese Patent Laid-Open No. 06-199647), as fermented products of animals and plants, aspergillus culture of green tea extract (Japanese Patent Laid-Open No. 2012-92100), lactic acid bacteria culture of pineapple juice and grape juice (Patent Document 2006-8566). As the hydrolyzate, a hydrolyzate of an extract of green tea (Japanese Unexamined Patent Publication No. 2011-256147) is exemplified. In addition to these, lychee seed extract, evening primrose extract, kiwi seed extract, warm ginkgo extract, ginkgo biloba extract, lactoferrin and the like, which are commercially available as having a tyrosinase inhibitory effect, can also be used. It can also be a lactic acid bacterium culture of pineapple fruit core, which is a tyrosinase activity inhibitor according to the present invention, or a lactic acid bacterium culture of pineapple fruit containing fruit core. One or more of these tyrosinase activity inhibitors are used in the whitening composition.
本願発明に係るチロシナーゼ活性阻害剤であるパイナップル果芯の乳酸菌による培養物（乳酸菌培養物）は、パイナップルの果芯を含む培地で乳酸菌を培養して得られる培養物である。培養物には、好気的条件下で培養して得られる培養物だけでなく、嫌気的あるいは微好気的条件下で培養して得られるいわゆる乳酸発酵物であってもよい。パイナップルの果芯は、パイナップルの果実から果皮を取り除いた後、可食部を得るために除去される芯の部分である。得られた果芯はそのまま培地として用いられ得るが、好ましくは果芯を破砕した破砕物が培地として用いられる。培地成分としては、果芯やその破砕物以外に少量の可食部や果汁が含まれることもあり、可食部や果汁を完全に取り除くことなく用いても差し支えない。さらには、本願発明ではパイナップルの果実の破砕物、すなわち、果芯部のみならず可食部、さらには果皮を含む果実を培地として用いて得られた乳酸菌培養物も用いることもできる。 The culture of pineapple fruit core lactic acid bacteria (lactic acid bacterium culture), which is the tyrosinase activity inhibitor according to the present invention, is a culture obtained by culturing lactic acid bacteria in a medium containing pineapple fruit core. The culture may be not only a culture obtained by culturing under aerobic conditions, but also a so-called lactic acid fermented product obtained by culturing under anaerobic or slightly aerobic conditions. The pineapple fruit core is the portion of the core that is removed to obtain an edible portion after removing the pericarp from the pineapple fruit. The obtained fruit core can be used as a medium as it is, but preferably a crushed product obtained by crushing the fruit core is used as a medium. As the medium component, a small amount of edible portion or fruit juice may be contained in addition to the fruit core and its crushed product, and the edible portion and fruit juice may be used without being completely removed. Further, in the present invention, a crushed product of pineapple fruit, that is, a lactic acid bacterium culture obtained by using not only the core portion but also the edible portion and the fruit containing the pericarp as a medium can also be used.
培養に用いられる乳酸菌は代謝産物として乳酸を産生する菌であればよく、ラクトバシラス属、ペディオコッカス属、ロイコノストック属、ストレプトコッカス属、ラクトコッカス属に属する細菌やビフィドバクテリウム属に属する細菌であり得る。例えば、ラクトバチルス・ブルガリクス、ラクトバチルス・ガセリ、ラクトバチルス・アシドフィラス、ラクトバチルス・カゼイ、ラクトバチルス・デルブルッキー、ラクトバチルス・プランタラム、ラクトバチルス・ブレビス、ラクトバチルス・ヘルベティカス、ラクトコッカス・ラクティス、ストレプトコッカス・サーモフィラス、ビフィドバクテリウム・ロンガム、ビフィドバクテリウム・ブレーベ、ビフィドバクテリウム・ビフィディウムが示される。培養には、１種のみの乳酸菌だけでなく２種以上の乳酸菌も用いられ得る。 The lactic acid bacterium used for culturing may be a bacterium that produces lactic acid as a metabolite, and is a bacterium belonging to the genus Lactobacillus, Pediococcus, Leuconostoc, Streptococcus, Lactococcus, or Bifidobacterium. Can be. For example, Lactobacillus bulgarix, Lactobacillus gasseri, Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus delbrucky, Lactobacillus plantarum, Lactobacillus brevis, Lactobacillus herveticas, Lactobacillus lactis, Streptococcus thermophilus, Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium bifidium are shown. Not only one type of lactic acid bacterium but also two or more types of lactic acid bacteria can be used for culturing.
培養は好気的条件から嫌気的条件の下で行われる。その培養条件、例えば、培養温度や培養時間等は当業者により適宜設定される。一例を挙げると、培地に乳酸菌を接種した後、３７℃で２４時間〜４８時間培養される。また、培地には必要に応じて、水、炭酸カルシウムなどのｐＨ調整剤、蛋白分解物などの窒素源、ブドウ糖、乳糖などの糖類、発酵促進剤などが加えられることもある。 Culturing is carried out under aerobic to anaerobic conditions. The culture conditions, for example, the culture temperature and the culture time, are appropriately set by those skilled in the art. As an example, after inoculating the medium with lactic acid bacteria, the medium is cultured at 37 ° C. for 24 to 48 hours. Further, if necessary, water, a pH adjuster such as calcium carbonate, a nitrogen source such as a hydrolyzate, sugars such as glucose and lactose, a fermentation accelerator and the like may be added to the medium.
得られた培養物はそのままチロシナーゼ活性阻害剤として用いられる場合もあれば、凍結乾燥などの方法により乾燥させた後に用いられる場合や濃縮してエキス状として用いられる場合もある。また、水、アルコールなどの極性溶媒やその他の非極性溶媒で抽出したエキスとしても用いられ得る。さらに必要に応じて、乳糖やデンプンなどの基材などが添加されたチロシナーゼ活性阻害剤として用いられる場合もある。 The obtained culture may be used as it is as a tyrosinase activity inhibitor, may be used after being dried by a method such as freeze-drying, or may be concentrated and used as an extract. It can also be used as an extract extracted with a polar solvent such as water or alcohol or another non-polar solvent. Further, if necessary, it may be used as a tyrosinase activity inhibitor to which a base material such as lactose or starch is added.
本願発明において用いられるプラセンタエキスは、動物の胎盤から得られたエキスであって、エキスの製造工程中において加熱処理、酵素処理又は加水分解処理の少なくとも１又は２以上の処理を経て得られるものである。これらの処理によってプラセンタエキスのチロシナーゼ阻害活性が失活し、チロシナーゼ阻害活性が低い、好ましくは活性がないとされるプラセンタエキスが用いられる。阻害活性が低いプラセンタエキスとは、具体的にはチロシナーゼ阻害活性が５０％以下、好ましくは３０％以下、より好ましくは２０％以下、さらに好ましくは１０％以下、より望ましくは５％以下のプラセンタエキスである。ここで言うチロシナーゼ阻害活性は、チロシナーゼ活性阻害剤と共に用いられるプラセンタエキスが液状の場合はその２００μLを試料溶液とし、粘稠状のプラセンタエキスなど液状以外の場合にはその粉末（蒸発固形分）の５mgを２００μLの水に溶解した液を試料溶液とし、下記実施例の記載の方法に従って測定した場合におけるチロシナーゼ阻害活性（％）を意味する。参考として述べると、２００μLの試料溶液（実施例中のプラセンタエキスＡ）は、特許文献１に記載されたプラセンタエキスの固形分と比較して、固形分換算で約３０倍の量である。また、１０％のチロシナーゼ阻害活性は実施例記載の方法ではアルブチン約５ｍＭ、５％のチロシナーゼ阻害活性は同様にアルブチン約２ｍＭの濃度に相当し、チロシナーゼ阻害活性が１０％以下であればチロシナーゼ阻害活性は実質的にないと言える。従って、本願発明においては２００μLの試料溶液でチロシナーゼ阻害活性１０％以下のプラセンタエキス、さらに１００μLの試料溶液で検出限界以下となるプラセンタエキスが好ましく用いられ得る。 The placenta extract used in the present invention is an extract obtained from the placenta of an animal, which is obtained through at least one or more treatments of heat treatment, enzyme treatment or hydrolysis treatment during the process of producing the extract. is there. By these treatments, the tyrosinase inhibitory activity of the placenta extract is inactivated, and the placenta extract having a low tyrosinase inhibitory activity, preferably inactive, is used. The placenta extract having low inhibitory activity is specifically a placenta extract having a tyrosinase inhibitory activity of 50% or less, preferably 30% or less, more preferably 20% or less, still more preferably 10% or less, and more preferably 5% or less. Is. The tyrosinase inhibitory activity referred to here is 200 μL of the placenta extract used together with the tyrosinase activity inhibitor as a sample solution when it is liquid, and its powder (evaporated solid content) when it is not liquid such as a viscous placenta extract. It means the tyrosinase inhibitory activity (%) when measured according to the method described in the following Examples using a solution prepared by dissolving 5 mg in 200 μL of water as a sample solution. As a reference, the amount of 200 μL of the sample solution (placenta extract A in the examples) is about 30 times as much as the solid content of the placenta extract described in Patent Document 1. In addition, 10% tyrosinase inhibitory activity corresponds to a concentration of about 5 mM arbutin and 5% tyrosinase inhibitory activity also corresponds to a concentration of about 2 mM arbutin by the method described in Examples, and if the tyrosinase inhibitory activity is 10% or less, the tyrosinase inhibitory activity. Can be said to be virtually nonexistent. Therefore, in the present invention, a placenta extract having a tyrosinase inhibitory activity of 10% or less in a 200 μL sample solution and a placenta extract having a detection limit or less in a 100 μL sample solution can be preferably used.
プラセンタエキスの由来は問われず、その由来はウシであり、ウマであり、ブタであり、ヒトであり、サメであり得る。また、エキスの製造工程も特に問われることはないが、製造段階において、加熱処理、酵素処理、加水分解処理の何れかの処理若しくはこれらの処理のうち２以上の処理を経ていることが必要とされる。このような処理によってチロシナーゼ阻害活性が著しく低下し、又はほとんど活性を認められなくなったプラセンタエキスでも、他のチロシナーゼ阻害活性剤と用いることでチロシナーゼ阻害活性が高められた美白用組成物を得られるからである。こうした加熱処理、酵素処理、加水分解処理の何れか１又は２以上の処理を経て得られたプラセンタエキスの製造方法の一例を挙げると、胎盤を凍結・融解を行い破砕した後、加熱抽出する方法がある。これらのプラセンタエキスは、各社から上市されているプラセンタエキスから選択され得る。 The origin of the placenta extract does not matter, and the origin can be cattle, horses, pigs, humans, and sharks. The process for producing the extract is not particularly limited, but it is necessary that the extract has undergone any of heat treatment, enzyme treatment, hydrolysis treatment, or two or more of these treatments at the production stage. Will be done. This is because even a placenta extract whose tyrosinase inhibitory activity is significantly reduced or whose activity is hardly observed by such treatment can be used in combination with another tyrosinase inhibitory agent to obtain a whitening composition having enhanced tyrosinase inhibitory activity. Is. An example of a method for producing placenta extract obtained through any one or more of the heat treatment, the enzyme treatment, and the hydrolysis treatment is a method in which the placenta is frozen and thawed, crushed, and then heat-extracted. There is. These placenta extracts can be selected from the placenta extracts marketed by each company.
本願発明に係る美白用組成物中のチロシナーゼ活性阻害剤やプラセンタエキスの量は適宜当業者によって定められる。チロシナーゼ活性阻害剤の活性やプラセンタエキスの固形分濃度や窒素含量、製造工程によっても異なるが、プラセンタエキスの配合量はチロシナーゼ阻害活性剤によるチロシナーゼ阻害活性を高めることができる量が加えられる。プラセンタエキスの添加量は、例えばチロシナーゼ阻害活性剤濃度の１／１０００であり、１／１００であり、１／１０であり、等倍であり、２倍であり、５倍であり、１０倍であり、１００倍の濃度であり得る。 The amount of the tyrosinase activity inhibitor or placenta extract in the whitening composition according to the present invention is appropriately determined by those skilled in the art. Although it depends on the activity of the tyrosinase activity inhibitor, the solid content concentration and nitrogen content of the placenta extract, and the production process, the amount of the placenta extract blended is an amount capable of enhancing the tyrosinase inhibitory activity of the tyrosinase inhibitory activator. The amount of placenta extract added is, for example, 1/1000, 1/100, 1/10, 1x, 2x, 5x, 10x the concentration of the tyrosinase inhibitor activator. Yes, it can be 100 times more concentrated.
本願発明に係る美白用組成物は、化粧用組成物であり、食品組成物であり、医薬組成物でもあり得る。その剤型も固形、液体等を問わない。当該美白用組成物はチロシナーゼ阻害剤やプラセンタエキスの他に、化粧品や食品、医薬品の製造に一般的に用いられる成分、例えば、油性成分や水、アルコールなどの製剤用基材成分や、チロシナーゼ活性阻害剤以外の美白剤、保湿剤、酸化防止剤、紫外線吸収剤、界面活性剤、増粘剤、着色剤、ビタミン類、保存料などの各種添加成分を必要に応じて含み得る。 The whitening composition according to the present invention may be a cosmetic composition, a food composition, or a pharmaceutical composition. The dosage form may be solid or liquid. In addition to the tyrosinase inhibitor and placenta extract, the whitening composition contains components generally used in the manufacture of cosmetics, foods, and pharmaceuticals, such as oily components, base components for preparations such as water and alcohol, and tyrosinase activity. Various additive components such as whitening agents, moisturizers, antioxidants, ultraviolet absorbers, surfactants, thickeners, colorants, vitamins, preservatives, etc. other than inhibitors may be contained as required.
本願発明によると、チロシナーゼ阻害活性が著しく低いかほとんど活性のないプラセンタエキスであっても、チロシナーゼ活性阻害剤と共に用いることでよりチロシナーゼ阻害活性の高い美白用組成物が得られる。特に、加熱処理、酵素処理、加水分解処理されたことによりチロシナーゼ阻害活性が失われ、実質的にチロシナーゼ阻害活性がないプラセンタエキスを用いることで、少なくともチロシナーゼ活性阻害剤の有するチロシナーゼ阻害活性とプラセンタエキスの有するチロシナーゼ阻害活性の総和よりも大きなチロシナーゼ阻害活性を有する組成物を、より好ましくはチロシナーゼ活性阻害剤のチロシナーゼ阻害活性よりも大きなチロシナーゼ阻害活性を有する組成物を得ることができる。また、廃棄物であるパイナップル果芯の有効利用も図ることができる。 According to the present invention, even a placenta extract having extremely low or almost no tyrosinase inhibitory activity can be used together with a tyrosinase activity inhibitor to obtain a whitening composition having higher tyrosinase inhibitory activity. In particular, by using a placenta extract that loses tyrosinase inhibitory activity due to heat treatment, enzyme treatment, and hydrolysis treatment and has substantially no tyrosinase inhibitory activity, at least the tyrosinase inhibitory activity and placenta extract possessed by the tyrosinase activity inhibitor A composition having a tyrosinase inhibitory activity larger than the sum of the tyrosinase inhibitory activities of the tyrosinase inhibitory activity, more preferably a composition having a tyrosinase inhibitory activity larger than the tyrosinase inhibitory activity of the tyrosinase activity inhibitor can be obtained. In addition, the pineapple fruit core, which is a waste, can be effectively used.
次に本願発明について下記の実施例に基づいて具体的に説明するが、本願発明は下記の実施例に限定されないのは言うまでもない。 Next, the present invention will be specifically described based on the following examples, but it goes without saying that the present invention is not limited to the following examples.
〔パイナップル果芯の乳酸菌培養物のチロシナーゼ阻害活性〕
表１に示す各種の乳酸菌を廃棄物として得られたパイナップル果芯の破砕物に接種して、３７℃で４６時間静置培養した。得られた培養物Ａ〜Ｆをそれぞれ凍結乾燥して得られた乾燥物５ｇに１００mLの純水を加え、１２,０００rpmで２０分間遠心分離して上清を得た。この上清を試料溶液として、下記の測定方法に従ってチロシナーゼ阻害活性を測定した。その結果を表１に示した。なお、実施例で用いられた乳酸菌は、国立研究開発法人、理化学研究所 バイオリソースセンター微生物材料開発室 (JCM)から入手したものである。
[Tyrosinase inhibitory activity of lactic acid bacteria culture of pineapple fruit core]
Various lactic acid bacteria shown in Table 1 were inoculated into crushed pineapple fruit cores obtained as waste, and statically cultured at 37 ° C. for 46 hours. 100 mL of pure water was added to 5 g of the obtained dried product obtained by freeze-drying each of the obtained cultures A to F, and the mixture was centrifuged at 12,000 rpm for 20 minutes to obtain a supernatant. Using this supernatant as a sample solution, the tyrosinase inhibitory activity was measured according to the following measurement method. The results are shown in Table 1. The lactic acid bacteria used in the examples were obtained from the Japan Collection of Microorganisms (JCM), BioResource Center, RIKEN.
（チロシナーゼ阻害活性の測定方法）
チロシナーゼの阻害活性は、マッシュルーム由来のチロシナーゼを用いて測定した。２０mM(pH６.５)のリン酸緩衝液に溶解した５mM L-DOPA ０.５mLに、試料溶液２００μLと水を加えて０.５mLに調製した溶液を加え、２５℃で５分間プレインキュベートののち、４００units/mlのマッシュルーム由来のチロシナーゼを１０μL添加し、２５℃で１５分間反応した。反応終了後４７５nmの吸光度を測定した。チロシナーゼ活性阻害率は次式により算出した。
阻害率（％）＝（１−（Ａ−Ａ０）／（Ｂ−Ｂ０））×１００
Ａ： 試料溶液を含む反応液の吸光度
Ｂ： 試料溶液を含まない反応液の吸光度
Ａ０： Ａのチロシナーゼの代わりにリン酸緩衝液を添加した反応液の吸光度
Ｂ０： Ｂのチロシナーゼの代わりにリン酸緩衝液を添加した反応液の吸光度
(Method for measuring tyrosinase inhibitory activity)
The inhibitory activity of tyrosinase was measured using tyrosinase derived from mushrooms. To 0.5 mL of 5 mM L-DOPA dissolved in 20 mM (pH 6.5) phosphate buffer, add 200 μL of the sample solution and a solution prepared to 0.5 mL by adding water, and pre-incubate at 25 ° C. for 5 minutes. , 400 units / ml of mushroom-derived tyrosinase was added, and the mixture was reacted at 25 ° C. for 15 minutes. After completion of the reaction, the absorbance at 475 nm was measured. The tyrosinase activity inhibition rate was calculated by the following formula.
Inhibition rate (%) = (1- (AA 0 ) / (BB 0 )) × 100
A: Absorbance of the reaction solution containing the sample solution B: Absorbance of the reaction solution not containing the sample solution A 0 : Absorbance of the reaction solution in which a phosphate buffer solution is added instead of the tyrosinase of A B 0 : Instead of the tyrosinase of B Absorbance of reaction solution to which phosphate buffer is added
次にパイナップルの果芯の破砕物に、乳酸菌Lactobacillus casei IAM1045を接種して３７℃で２日間静置培養した。得られた培養物について、試料溶液の使用量を変えて上記方法と同様にしてチロシナーゼ阻害活性を測定した。また、参考として、パイナップル果汁についても同様に培養させた後、チロシナーゼ阻害活性を測定し、果芯培養物と比較した。それらの結果を図１及び図２に示した。なお、振盪培養したところ、静置培養と同様な結果が得られた。 Next, the crushed pineapple fruit core was inoculated with the lactic acid bacterium Lactobacillus casei IAM1045 and statically cultured at 37 ° C. for 2 days. For the obtained culture, the tyrosinase inhibitory activity was measured in the same manner as in the above method by changing the amount of the sample solution used. For reference, pineapple juice was also cultured in the same manner, and then the tyrosinase inhibitory activity was measured and compared with the fruit core culture. The results are shown in FIGS. 1 and 2. When the culture was shaken, the same results as the static culture were obtained.
表１に示すようにパイナップル果芯の培養物Ａ〜Ｆは２０時間又は４６時間の培養でほぼ５０％以上のチロシナーゼ阻害活性を示し、培養することでチロシナーゼ阻害活性が増加した。また、各培養物は容量依存性を示し、２００μLの使用量で強いチロシナーゼ阻害活性を示した（図１参照）。一方、果汁を用いて同様に培養した場合には、図２に示すように果芯を用いた場合に比べて、チロシナーゼ阻害活性が出現するまでにより多くの時間を必要とした。 As shown in Table 1, the pineapple fruit core cultures A to F showed approximately 50% or more of tyrosinase inhibitory activity after 20 hours or 46 hours of culture, and the tyrosinase inhibitory activity was increased by culturing. In addition, each culture showed volume dependence and showed strong tyrosinase inhibitory activity at a dose of 200 μL (see FIG. 1). On the other hand, when the same culture was carried out using fruit juice, it took more time for the tyrosinase inhibitory activity to appear as compared with the case where the fruit core was used as shown in FIG.
〔プラセンタエキスのチロシナーゼ阻害活性〕
市販されているウマ胎盤由来のプラセンタエキス２種類（ロットの異なるプラセンタエキスＡ及びプラセンタエキスＢ）を使用して、実施例１と同様にしてチロシナーゼ阻害活性を調べた。それらの結果を表２及び図３に示した。これらのプラセンタエキスは、ウマの胎盤を、凍結、融解したものに冷水を加えた後、加熱抽出、脱臭操作して得られたエキスである。
[Tyrosinase inhibitory activity of placenta extract]
Using two types of commercially available placenta extracts derived from horse placenta (placenta extract A and placenta extract B from different lots), the tyrosinase inhibitory activity was examined in the same manner as in Example 1. The results are shown in Table 2 and FIG. These placenta extracts are extracts obtained by adding cold water to frozen and thawed horse placenta, and then heat-extracting and deodorizing.
図３は、プラセンタエキスＡによるチロシナーゼ阻害活性について実施例１に記載の方法に準じて調べた結果であるが、１００μL以下の添加量では全く活性を示さなかった。さらに、プラセンタエキスＡ，Ｂの各２００μLを用いて、各プラセンタエキスのチロシナーゼ阻害活性を測定した。表２に示したように、プラセンタエキスＡ,Ｂの阻害活性はそれぞれ１０％以下であり、プラセンタエキスＡ，Ｂにはほとんどチロシナーゼ阻害活性は認められなかいと言えるか、極めて低い活性であった。 FIG. 3 shows the results of examining the tyrosinase inhibitory activity of placenta extract A according to the method described in Example 1, but no activity was shown when the amount added was 100 μL or less. Furthermore, 200 μL each of placenta extracts A and B was used to measure the tyrosinase inhibitory activity of each placenta extract. As shown in Table 2, the inhibitory activity of placenta extracts A and B was 10% or less, respectively, and it can be said that almost no tyrosinase inhibitory activity was observed in placenta extracts A and B, or the activity was extremely low.
〔パイナップル果芯の乳酸菌培養物とプラセンタエキスの併用効果〕
次に、実施例１で得られた果芯の乳酸菌培養物とプラセンタエキスの併用効果について調べた。図４に示す割合となるように、実施例１で得られた果芯の乳酸菌培養物ＡとプラセンタエキスＡを混合して試料溶液を調整した後、実施例１に記載した方法に準じて、チロシナーゼ阻害活性を測定した。その結果を図４に示す。また、実施例１で得られた果芯の乳酸菌培養物ＢとプラセンタエキスＡ，Ｂを混合した場合についても同様にチロシナーゼ阻害活性を測定した。その結果を図５に示した。
[Effect of combined use of pineapple fruit core lactic acid bacteria culture and placenta extract]
Next, the combined effect of the lactic acid bacterium culture of the fruit core obtained in Example 1 and the placenta extract was investigated. The sample solution was prepared by mixing the lactic acid bacterium culture A of the fruit core obtained in Example 1 and the placenta extract A so as to have the ratio shown in FIG. 4, and then according to the method described in Example 1. The tyrosinase inhibitory activity was measured. The result is shown in FIG. In addition, the tyrosinase inhibitory activity was measured in the same manner when the lactic acid bacterium culture B of the fruit core obtained in Example 1 and the placenta extracts A and B were mixed. The result is shown in FIG.
その結果、得られた乳酸菌培養物の試料溶液に対して、等量のプラセンタエキスを加えた場合には、その活性は約２倍以上に増加した。チロシナーゼ阻害活性がほとんど認められないプラセンタエキスを用いて、用いたパイナップル果芯の乳酸菌培養物のチロシナーゼ阻害活性以上のチロシナーゼ阻害活性が得られることが確認された。 As a result, when an equal amount of placenta extract was added to the sample solution of the obtained lactic acid bacterium culture, its activity increased more than twice. It was confirmed that the placenta extract, which has almost no tyrosinase inhibitory activity, can be used to obtain tyrosinase inhibitory activity equal to or higher than that of the lactic acid bacterium culture of the pineapple fruit core used.
ちなみに、上記で用いられたプラセンタエキスの固形分（蒸発残留分）は、エキスＡが１.４５w/v％、エキスＢが２.２３w/v％であった。また、液体クロマトグラフにより、エキス中の遊離のアミノ酸含量及びチロシン含量を測定したところ、遊離のアミノ酸含量はそれぞれ１.４９mg/mL、１.２３mg/mLであり、チロシン含量はそれぞれ０.０３mg/mL、０.０４mg/mLであった。このように、固形分中の遊離チロシン含量が低い、例えば特許文献３の比較例にあるような１０％以下、さらには５％以下のプラセンタエキスや、プラセンタエキスＡ、Ｂのように特許文献３の実施例にあるような０.３％以下などのような遊離チロシン含量が低いプラセンタエキスであっても極めて低いチロシナーゼ阻害活性しか得られない場合があることが確認された。また、特許文献３に記載の方法に準じて、これらのプラセンタエキスのタンパク量を求めたところ、分子量３０００を越えるタンパク量はいずれも３０?６０mg/100mL（固形分に対して２?３％）程度であった。これらのことから、プラセンタエキスＡ,Ｂは何れも特許文献３の比較例にあるプラセンタエキスと同様に、酵素処理や加熱処理等によりタンパクやペプチド等が分解してチロシナーゼ阻害活性が失われていると推測される。そして、加熱処理や酵素処理等により低いチロシナーゼ阻害活性しか示されないプラセンタエキスであっても、例えそのチロシン含有量が低いとしても、他のチロシナーゼ活性阻害剤と組み合わせることで、より効果の高い美白用組成物を得ることができると言える。 Incidentally, the solid content (evaporation residue) of the placenta extract used above was 1.45 w / v% for extract A and 2.23 w / v% for extract B. Moreover, when the free amino acid content and the tyrosine content in the extract were measured by a liquid chromatograph, the free amino acid content was 1.49 mg / mL and 1.23 mg / mL, respectively, and the tyrosine content was 0.03 mg / mL, respectively. It was mL, 0.04 mg / mL. As described above, the placenta extract having a low free tyrosine content in the solid content, for example, 10% or less as in the comparative example of Patent Document 3 and further 5% or less, and Placenta Extracts A and B such as Placenta Extract 3 It was confirmed that even a placenta extract having a low free tyrosine content, such as 0.3% or less as in the example of, may obtain extremely low tyrosinase inhibitory activity. Further, when the protein amount of these placenta extracts was determined according to the method described in Patent Document 3, the amount of protein having a molecular weight exceeding 3000 was 30 to 60 mg / 100 mL (2 to 3% with respect to the solid content). It was about. From these facts, in both placenta extracts A and B, like the placenta extract in the comparative example of Patent Document 3, proteins and peptides are decomposed by enzyme treatment, heat treatment, etc., and the tyrosinase inhibitory activity is lost. It is presumed. And even if the placenta extract shows only low tyrosinase inhibitory activity by heat treatment or enzyme treatment, even if its tyrosine content is low, it is more effective for whitening by combining with other tyrosinase activity inhibitors. It can be said that the composition can be obtained.
次に下記に示す処方例に従って各種の美白用組成物を作製したところ、プラセンタエキスを加えない場合に比べてチロシナーゼ阻害活性が高められ、より強い美白効果が期待される美白用組成物が得られた。なお、処方例１，２では、実施例１で得られたパイナップル果芯の乳酸菌培養物を凍結乾燥し、２０倍量の精製水を加えて抽出し、遠心分離して得られた上清画分を、乳酸発酵パイナップル果芯エキスとして使用し、処方例４では、実施例１で得られたパイナップル果芯の乳酸発酵培養物を滅菌処理後、凍結乾燥したものを使用した。また、プラセンタエキスＡ末は、プラセンタエキスＡを凍結乾燥したものである。
（処方例１：化粧水）
下記組成の化粧水を常法により製造した。
プラセンタエキスＡ ０．０２ｇ
乳酸発酵パイナップル果芯エキス ０．０２ｇ
ヒアルロン酸ナトリウム ０．０２ｇ
グルタチオン ０．００１ｇ
１，３−ブチレングリコール ８ｇ
ペンチレングリコール ３．５ｇ
キサンタンガム ０．５ｇ
香料 ０．１ｇ
精製水 残部（全量を１００ｇとする）
Next, when various whitening compositions were prepared according to the formulation examples shown below, the tyrosinase inhibitory activity was enhanced as compared with the case where the placenta extract was not added, and a whitening composition expected to have a stronger whitening effect was obtained. It was. In Formulation Examples 1 and 2, the lactic acid bacterium culture of the pineapple fruit core obtained in Example 1 was freeze-dried, extracted by adding 20 times the amount of purified water, and centrifuged to obtain a supernatant image. The minutes were used as a lactic acid fermented pineapple fruit core extract, and in Formulation Example 4, the lactic acid fermented culture of the pineapple fruit core obtained in Example 1 was sterilized and then freeze-dried. The placenta extract A powder is freeze-dried placenta extract A.
(Prescription example 1: Toner)
A lotion having the following composition was produced by a conventional method.
Placenta Extract A 0.02g
Lactic acid fermented pineapple fruit core extract 0.02g
Sodium hyaluronate 0.02g
Glutathione 0.001g
1,3-butylene glycol 8g
Pentylene glycol 3.5g
Xanthan gum 0.5g
Fragrance 0.1g
Purified water balance (total amount is 100 g)
（処方例２：ゲルクリーム）
下記組成のゲルクリームを常法により製造した。
プラセンタエキスＢ ０．３ｇ
乳酸発酵パイナップル果芯エキス ０．１ｇ
グリセリン ６ｇ
ミリスチン酸イソプロピル ３ｇ
ジメチコン ３ｇ
シクロメチコン １ｇ
１，３−ブチレングリコール ０．７ｇ
ヒアルロン酸ナトリウム ０．０３ｇ
スクワラン ０．５ｇ
オリーブ油 ０．２ｇ
エタノール ０．２ｇ
精製水 残部（全量を１００ｇとする）
(Prescription example 2: Gel cream)
A gel cream having the following composition was produced by a conventional method.
Placenta extract B 0.3g
Lactic acid fermented pineapple fruit core extract 0.1g
Glycerin 6g
Isopropyl myristate 3g
Dimethicone 3g
Cyclomethicone 1g
1,3-butylene glycol 0.7g
Sodium hyaluronate 0.03g
Squalene 0.5g
Olive oil 0.2g
Ethanol 0.2g
Purified water balance (total amount is 100 g)
（処方例３：エッセンス）
下記組成のエッセンスを常法により製造した。
プラセンタエキスＡ ２ｇ
月見草エキス ０．５ｇ
１，２−ヘキサンジオール ０．０４ｇ
カプリリルグリコール ０．０４ｇ
精製水 残部（全量を１００ｇとする）
(Prescription example 3: Essence)
An essence having the following composition was produced by a conventional method.
Placenta extract A 2g
Evening primrose extract 0.5g
1,2-Hexanediol 0.04g
Caprylyl glycol 0.04g
Purified water balance (total amount is 100 g)
（処方例４：清涼飲料水）
下記組成の清涼飲料水を常法により製造した。
プラセンタエキスＡ末 １０ｇ
乳酸発酵パイナップル果芯エキス ３０ｇ
グラニュー糖 １２ｇ
コラーゲン １０ｇ
ヒハツエキス ０．１５ｇ
ローヤルゼリー ０．０５ｇ
ビタミンＣ ０．７５ｇ
クエン酸 ０．０７５ｇ
香料 ０．３ｍｌ
精製水 残部（全量を１００ｇとする）
(Prescription example 4: Soft drink)
A soft drink having the following composition was produced by a conventional method.
Placenta extract A powder 10g
Lactic acid fermented pineapple fruit core extract 30g
Granulated sugar 12g
Collagen 10g
Hihatsu extract 0.15g
Royal jelly 0.05g
Vitamin C 0.75g
Citric acid 0.075g
Fragrance 0.3ml
Purified water balance (total amount is 100 g)
（処方例５：錠剤型サプリメント）
下記組成の混合物を打錠し、常法により錠剤型サプリメントを製造した。
プラセンタエキスＢ末 ３５０ｇ
キウイ種子エキス １７５ｇ
ヒドロキシプロピルメチルセルロース ６０ｇ
(Prescription example 5: Tablet-type supplement)
A mixture having the following composition was tableted to prepare a tablet-type supplement by a conventional method.
Placenta extract B powder 350g
Kiwi seed extract 175g
Hydroxypropyl Methyl Cellulose 60g
本願発明は、プラセンタエキスを含む新たな美白用組成物の製造に寄与する。 The present invention contributes to the production of a new whitening composition containing placenta extract.
Claims (3)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2018002679A JP6792270B2 (en) | 2018-01-11 | 2018-01-11 | Whitening composition |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP2018002679A JP6792270B2 (en) | 2018-01-11 | 2018-01-11 | Whitening composition |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JP2019119724A JP2019119724A (en) | 2019-07-22 |
| JP6792270B2 true JP6792270B2 (en) | 2020-11-25 |
Family
ID=67306107
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP2018002679A Active JP6792270B2 (en) | 2018-01-11 | 2018-01-11 | Whitening composition |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP6792270B2 (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP6944606B1 (en) * | 2021-03-30 | 2021-10-06 | 新日本製薬株式会社 | Manufacturing method of cosmetic composition |
Family Cites Families (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS60202806A (en) * | 1984-03-26 | 1985-10-14 | Sansho Seiyaku Kk | Whitening cosmetic |
| JPH07173024A (en) * | 1993-12-16 | 1995-07-11 | Shiseido Co Ltd | Skin external preparation |
| JP2002179523A (en) * | 2000-12-15 | 2002-06-26 | Ichimaru Pharcos Co Ltd | Cosmetic composition containing extract of swine placenta |
| JP2002187812A (en) * | 2000-12-20 | 2002-07-05 | Ichimaru Pharcos Co Ltd | Cosmetic composition containing horse placenta extract |
| JP2006008566A (en) * | 2004-06-24 | 2006-01-12 | Ichimaru Pharcos Co Ltd | Cosmetic agent containing lactic acid bacteria fermentation product of fruit juice as active ingredient and its application |
| JP2010030946A (en) * | 2008-07-29 | 2010-02-12 | Asuka Corporation:Kk | Whitening cosmetic |
| JP2010138147A (en) * | 2008-12-15 | 2010-06-24 | Op Bio Factory Co Ltd | Melanogenesis inhibitor, antioxidant, anti-inflammatory agent, external preparation for skin, and food and beverage |
| JP6505392B2 (en) * | 2014-08-19 | 2019-04-24 | プランドゥシー・メディカル株式会社 | Moisturizer and external preparation for skin containing the same |
| JP6346828B2 (en) * | 2014-08-22 | 2018-06-20 | イビデン株式会社 | Placenta extract |
| CN107648151A (en) * | 2017-09-23 | 2018-02-02 | 浙江长生鸟健康科技股份有限公司 | A kind of pearl sunscreen capsule and preparation method thereof |
-
2018
- 2018-01-11 JP JP2018002679A patent/JP6792270B2/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| JP2019119724A (en) | 2019-07-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| TWI494101B (en) | Improved biological effects of compositions comprising rosmarinic acid | |
| US8088369B2 (en) | Anti-wrinkle agent | |
| US20120301452A1 (en) | Probiotic microorganisms as active agents for enhancing the radiance of the skin's complexion | |
| KR101132392B1 (en) | Fermented Placenta Compositin and Use Thereof | |
| CN104277998A (en) | Use and methods for preventing and/or treating oral malodour | |
| JP2006094853A (en) | Fermented product obtained from garlic | |
| JP2017202990A (en) | Cosmetics and food and drink composition | |
| JP4286513B2 (en) | Anti-aging composition | |
| JP2015096476A (en) | Composition for collagen production promotion, composition for collagen absorption promotion, and composition for anti-obesity | |
| JP2022078231A (en) | Skin external preparations | |
| JP6482930B2 (en) | Skin cosmetics and foods and drinks | |
| JP2010540623A (en) | Soybean fermented polymer substance mixed with folic acid and composition containing the same | |
| CN115025129B (en) | Composition containing lactobacillus and use thereof for preventing and/or improving skin aging | |
| JP2017048244A (en) | Composition for collagen production promotion, composition for collagen absorption promotion, and composition for anti-obesity | |
| KR20160129453A (en) | Cosmetic composition for skin winkle improvement containing Fermented extract of horse placenta and horse oil | |
| JP6792270B2 (en) | Whitening composition | |
| KR20130099104A (en) | Skin collagen production-promoting agent | |
| TWI679982B (en) | Use of lactic acid bacteria fermented material for producing inhibitor of pentosine | |
| TWI492768B (en) | Cell protective agent | |
| JP2004196664A (en) | Humectant and cosmetic comprising the same | |
| JP6090854B2 (en) | Composition for promoting collagen production | |
| TWI794766B (en) | A composition containing lactobacillus spp. and its use for preventing and/or improving anti-aging of skin | |
| JP6209580B2 (en) | Ultraviolet ray obstruction reducing agent, method for producing ultraviolet ray obstruction reducing agent, medium additive, microorganism diluent additive, medium, microorganism diluent, external preparation and cosmetics | |
| JP2021187793A (en) | Agent for proliferation of mesenchymal stem cells | |
| JP2004107242A (en) | Kit for improving dry skin |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20200306 |
|
| A871 | Explanation of circumstances concerning accelerated examination |
Free format text: JAPANESE INTERMEDIATE CODE: A871 Effective date: 20200306 |
|
| A975 | Report on accelerated examination |
Free format text: JAPANESE INTERMEDIATE CODE: A971005 Effective date: 20200318 |
|
| A977 | Report on retrieval |
Free format text: JAPANESE INTERMEDIATE CODE: A971007 Effective date: 20200709 |
|
| A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20200804 |
|
| A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20200930 |
|
| TRDD | Decision of grant or rejection written | ||
| A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20201027 |
|
| A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20201028 |
|
| R150 | Certificate of patent or registration of utility model |
Ref document number: 6792270 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
| R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |