JP3625501B2 - Method for producing raw material for skin-applied skin and cell-applied skin external agent - Google Patents
Method for producing raw material for skin-applied skin and cell-applied skin external agent Download PDFInfo
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- JP3625501B2 JP3625501B2 JP24316994A JP24316994A JP3625501B2 JP 3625501 B2 JP3625501 B2 JP 3625501B2 JP 24316994 A JP24316994 A JP 24316994A JP 24316994 A JP24316994 A JP 24316994A JP 3625501 B2 JP3625501 B2 JP 3625501B2
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- skin
- cell
- external preparation
- raw material
- skin external
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- Medicines Containing Plant Substances (AREA)
- Cosmetics (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Description
【0001】
【産業上の利用分野】
本発明は、安全性が高く、皮膚の活性を高める細胞賦活用皮膚外用剤原料の製造方法及び細胞賦活用皮膚外用剤に関する。
【0002】
【従来の技術】
一般に、皮膚外用剤に求められる効能は数多くあるが、その一つに細胞賦活作用がある。細胞を賦活することによって、創傷治癒、火傷などの改善、または、しもやけ、あかぎれ、肌荒れなどの予防効果がある。
これらの薬効成分として、従来より、アロエエキス、人参エキス、菌発酵代謝物などが利用されてきたが、充分な効果が必ずしも得られていない点に未だ課題がある。
【0003】
【発明が解決しようとする課題】
本発明の目的は、上記従来の課題を解決するものであり、安全性が高く、皮膚の活性を高める細胞賦活用皮膚外用剤原料の製造方法及び細胞賦活用皮膚外用剤を提供することにある。
【0004】
【課題を解決するための手段】
本発明者らは、前記の課題を解決するために、鋭意検討した結果、より安全性の高い天然物由来の細胞賦活作用がある物質を得ることに成功し、本発明を完成するに至ったのである。
すなわち、本発明の細胞賦活用皮膚外用剤原料の製造方法は、米ぬかと大豆ペプチドとを枯草菌〔バチルス・ズブチリス(Bacillus subtilis)〕で発酵させた後、精製処理して細胞賦活作用を有する皮膚外用剤原料を製造することを特徴とする。
枯草菌は、納豆菌〔バチルス・ナットウ(Bacillus natto)〕であることが好ましい。
また、本発明の細胞賦活用皮膚外用剤は、上記の方法で製造した細胞賦活用皮膚外用剤原料を有効成分として含有することを特徴とする。
【0005】
【作用】
本発明の製造方法では、天然物から得られる米ぬかと大豆ペプチドとを出発原料とし、しかも、入手も容易な枯草菌により発酵させた後、精製処理するものであるので、細胞賦活作用を有する皮膚外用剤原料を安全に且つ容易に製造することができることとなる。
上記製造方法で得られた原料は、発酵中に生成蓄積されるビタミン類、アミノ酸類などを含有することとなり、該ビタミン類、アミノ酸類などが総合的に且つ相乗的なものとなった結果、強い細胞賦活作用、他にも皮膚外用剤としての効用を与えるものと推察される(この点等については、更に実施例等で詳しく説明する)。
【0006】
以下、本発明の内容を説明する。
本発明の細胞賦活用皮膚外用剤原料の製造方法は、米ぬかと大豆ペプチドとを枯草菌で発酵させた後、精製処理することにより細胞賦活作用を有する皮膚外用剤原料が製造される。
米ぬか、大豆ペプチドは、市販の米ぬか、大豆ペプチドを使用することができ、その配合割合は、米ぬか:大豆ペプチド100:1〜100:20部程度用いられる。これらの範囲にある場合には菌の生育に最も好ましいものとなる。
発酵に用いる枯草菌(Bacillus subtilis)は、好気性胞子形成細菌の代表的種類であり、この枯草菌としては、安全性が保証されているものであれば特に限定されないが、入手の容易性、コスト的にみて、納豆菌(Bacillus natto)の使用が好ましい。
【0007】
発酵させるのに、最適なpHや栄養条件等を得るためにグルコース等の糖類、炭酸水素ナトリウム等の塩類、プロテアーゼ等の酵素、水(精製水)等の添加も必要である。
本発明において精製処理は、上記発酵後の液を、圧搾し、ろ過することにより行われる。また、必要に応じて発酵後、圧搾し、ろ過し、次いで、活性炭等により脱臭・脱色処理、及び/又は、沈殿物の除去処理を行った後、再度ろ過することにより処理してもよい。
【0008】
また、本発明の細胞賦活用皮膚外用剤は、上記の方法で製造した細胞賦活用皮膚外用剤原料を有効成分とするものであり、該皮膚外用剤原料と、他の皮膚外用剤用の配合剤とを配合することにより得られる。すなわち、この皮膚外用剤原料を他の皮膚外用剤用の配合剤、例えば、スクワラン、ホホバ油等の液状油、ミツロウ、セチルアルコール等の固体油、各種の活性剤、グリセリン、1,3ーブチレングリコール等の保湿剤や各種薬剤等を配合して様々な剤形の細胞賦活用皮膚外用剤、例えば、ローション、クリーム、乳液、パック等、目的に応じて種々の利用形態の細胞賦活用皮膚外用剤などに調製することができる。
【0009】
【実施例】
以下に、本発明の細胞賦活用皮膚外用剤原料の製造例、実際の利用方法である実施例を記載するが、本発明はこれらの製造例及び実施例によって何ら限定されるものではない。
【0010】
〔製造例1〕
米ぬか3.00kg、リン酸一水素ナトリウム1.00kg、大豆ペプチド0.40kg、炭酸水素ナトリウム4.50kg、グルコース0.50kg、アルカリ性プロテアーゼ0.01kg、精製水50.00kgを混合した後、pH8.8〜9.0に調整した培地で納豆菌(バチルス ナットウ)を42℃、48時間培養した後、該培養液を、圧搾、ろ過、活性炭による脱色脱臭後、再度ろ過することによって皮膚外用剤原料を製造した。
【0011】
〔実施例1(クリームの調製)〕
下記諸成分からなるAとBとをそれぞれ70℃まで加温し、次いで、BにAを撹拌しつつ徐々に加えた後、ゆっくりと撹拌しつつ30℃まで冷却してクリームを調製した。
【0012】
〔細胞賦活試験〕
(試験方法)
24ウェルマルチプレートにFGM培地1mlを分注し、人正常繊維芽細胞を1万個播種して、2日間炭酸ガス培養器中で前培養した。
培地を除いた後、無血清MEM培地1mlで2回洗浄後、硫酸カナマイシン(100mg/l)のFBM培地(ホルモン未添加基本培地)1mlを加え、2日間培養した後、培地を除きFBM培地(ホルモン未添加基本培地)で各濃度に調整した実施例1を加えて6日間培養した。この間、3日目と5日目に培地交換を行った。
【0013】
MTT〔3−(4,5−ジメチル−2−チアゾリル)−2,5−ジフェニルテトラゾリウム ブロマイド〕溶液(5mg/mlリン酸緩衝液)0.1mlを加えて4時間培養後、20%ドデシル硫酸ナトリウム0.01N塩酸溶液1mlを加えて色素を溶解し、570nmの吸光度を測定した。対照として、実施例1を入れないもの(対照)、また、現在使用されている化粧品原料(胎盤抽出物、乳酸発酵代謝物)と比較した。その結果を下記表1に示す。なお、評価基準は、数値が大きい程、細胞賦活性が高いことを示し、対照を1として評価した。
【0014】
【表1】
【0015】
上記表1の結果から明らかなように、製造例の原料は、現在使用されている化粧品原料(胎盤抽出物、乳酸発酵代謝物)よりも低濃度(1/20)にもかかわらず、細胞賦活活性が高いことが判明した。
【0016】
次に、上記実施例1で調製したクリームを使用して下記のテストを行った。
(使用テスト)
女性6名の顔面を左右に分け、一方に、実施例のクリームを、他方には比較例のクリームを毎日、1回以上使用してもらって、3カ月後に、、肌荒れ防止、肌のつや及び肌のはりについて評価した。なお、比較例は実施例より製造例を精製水に代えたものである(比較例)。
【0017】
評価は、下記の評価基準により評価し、その結果をまとめたのが下記の表2である。
(評価基準)
実施例の方が非常によい 3
実施例の方がかなりよい 2
実施例の方がややよい 1
差がない 0
比較例の方がややよい −1
比較例の方がかなりよい −2
比較例の方が非常によい −3
【0018】
【表2】
【0019】
上記使用テスト(表2)から明らかなように、本発明の皮膚外用剤(クリーム)は、肌荒れ防止、肌のつや及び肌のはりに有効であり、安全性が高く、皮膚の活性を高める効果を有することが判った。
【0020】
【発明の効果】
本発明の製造方法では、天然物から得られる米ぬかと大豆ペプチドを出発原料とし、しかも、入手も容易な枯草菌により発酵させた後、精製処理することにより細胞賦活作用を有する皮膚外用剤原料が製造されることとなるので、大型の設備を必要とすることなく、安全に且つ容易に製造することができる。
本発明の細胞賦活用皮膚外用剤では、安全性が高く、皮膚の活性を高めることができ、非常に有用なものである。[0001]
[Industrial application fields]
The present invention relates to a method for producing a cell-utilizing skin external preparation material that is highly safe and enhances skin activity, and a cell-utilizing skin external preparation.
[0002]
[Prior art]
In general, there are many effects required for external preparations for skin, one of which is cell activation. By activating cells, there is an improvement effect on wound healing, burns, etc., or a preventive effect such as moistness, redness, and rough skin.
As these medicinal ingredients, aloe extract, ginseng extract, fungal fermentation metabolites and the like have been conventionally used, but there is still a problem in that sufficient effects are not necessarily obtained.
[0003]
[Problems to be solved by the invention]
An object of the present invention is to solve the above-described conventional problems, and to provide a method for producing a cell-utilizing skin external preparation material that is highly safe and enhances skin activity, and a cell-utilizing skin external preparation. .
[0004]
[Means for Solving the Problems]
As a result of intensive studies to solve the above-mentioned problems, the present inventors succeeded in obtaining a safer substance having a cell activation effect derived from a natural product, and completed the present invention. It is.
That is, the method for producing a cell-stimulating skin external preparation raw material of the present invention is a method of fermenting rice bran and soybean peptide with Bacillus subtilis (Bacillus subtilis), and then purifying the skin to have a cell-activating effect. A raw material for external preparation is produced.
The Bacillus subtilis is preferably Bacillus natto [Bacillus natto].
Moreover, the cell utilization skin external preparation of this invention contains the cell utilization skin external preparation raw material manufactured by said method as an active ingredient.
[0005]
[Action]
In the production method of the present invention, since rice bran obtained from a natural product and soybean peptide are used as starting materials, and further fermented with Bacillus subtilis, which is easily available, and then purified, skin having a cell activation effect An external preparation raw material can be manufactured safely and easily.
The raw material obtained by the above production method contains vitamins and amino acids that are produced and accumulated during fermentation, and as a result, the vitamins and amino acids are comprehensive and synergistic. It is presumed that it has a strong cell activation effect and other effects as an external preparation for skin (this point will be further described in detail in Examples and the like).
[0006]
The contents of the present invention will be described below.
In the method for producing a cell-utilizing skin external preparation raw material of the present invention, a rice external preparation and a soybean peptide are fermented with Bacillus subtilis and then purified to produce a skin external preparation raw material having a cell activating effect.
As the rice bran and soybean peptide, commercially available rice bran and soybean peptide can be used, and the blending ratio thereof is about rice bran: soybean peptide 100: 1 to 100: 20 parts. When it is within these ranges, it is most preferable for the growth of bacteria.
Bacillus subtilis used for fermentation is a representative type of aerobic spore-forming bacteria, and as such Bacillus subtilis is not particularly limited as long as safety is guaranteed, In view of cost, the use of Bacillus natto is preferable.
[0007]
In order to obtain an optimum pH and nutrient conditions for fermentation, addition of saccharides such as glucose, salts such as sodium bicarbonate, enzymes such as protease, water (purified water) and the like is also necessary.
In this invention, a refinement | purification process is performed by pressing and filtering the liquid after the said fermentation. Moreover, you may process by squeezing and filtering after fermentation as needed, and then filtering again after performing a deodorizing and decoloring process with activated carbon etc. and / or a deposit removal process.
[0008]
Further, the cell-utilizing skin external preparation of the present invention comprises the cell-utilizing skin external preparation raw material produced by the above method as an active ingredient, and the skin external preparation raw material and a combination for other skin external preparations It is obtained by blending with an agent. That is, this skin external preparation raw material is used as a compounding agent for other skin external preparations, for example, liquid oil such as squalane and jojoba oil, solid oil such as beeswax and cetyl alcohol, various active agents, glycerin, and 1,3-butylene. Cell use skin external preparations with various dosage forms by blending moisturizers such as glycol and various drugs, for example, lotions, creams, emulsions, packs, etc. It can be prepared as an agent.
[0009]
【Example】
Below, although the manufacture example of the cell utilization skin external preparation raw material of this invention and the Example which is an actual utilization method are described, this invention is not limited at all by these manufacture examples and Examples.
[0010]
[Production Example 1]
After mixing rice bran 3.00 kg, sodium monohydrogen phosphate 1.00 kg, soybean peptide 0.40 kg, sodium bicarbonate 4.50 kg, glucose 0.50 kg, alkaline protease 0.01 kg, purified water 50.00 kg, pH 8. After culturing Bacillus natto (Bacillus natto) for 48 hours at 42 ° C. in a medium adjusted to 8 to 9.0, the culture solution is compressed, filtered, decolored and deodorized with activated carbon, and then filtered again to obtain a raw material for external preparations for skin Manufactured.
[0011]
[Example 1 (Preparation of cream)]
A and B composed of the following components were each heated to 70 ° C., and then A was gradually added to B while stirring, and then cooled to 30 ° C. with slow stirring to prepare a cream.
[0012]
[Cell activation test]
(Test method)
1 ml of FGM medium was dispensed into a 24-well multiplate, 10,000 human normal fibroblasts were seeded, and precultured in a carbon dioxide incubator for 2 days.
After removing the medium, the cells were washed twice with 1 ml of serum-free MEM medium, 1 ml of KBM (100 mg / l) FBM medium (basic medium without hormones) was added, and cultured for 2 days. Then, the medium was removed and the FBM medium ( Example 1 adjusted to each concentration with a hormone-free basic medium) was added and cultured for 6 days. During this period, the medium was changed on the third and fifth days.
[0013]
MTT [3- (4,5-dimethyl-2-thiazolyl) -2,5-diphenyltetrazolium bromide] solution (5 mg / ml phosphate buffer) 0.1 ml was added and cultured for 4 hours, then 20% sodium dodecyl sulfate. The dye was dissolved by adding 1 ml of 0.01N hydrochloric acid solution, and the absorbance at 570 nm was measured. As a control, Example 1 was not included (control) and compared with cosmetic raw materials currently used (placenta extract, lactic acid fermentation metabolite). The results are shown in Table 1 below. In addition, evaluation criteria showed that cell activation was so high that a numerical value was large, and it evaluated as a control | contrast.
[0014]
[Table 1]
[0015]
As is clear from the results of Table 1 above, the raw material of the production example is activated even though the concentration (1/20) is lower than the cosmetic raw materials currently used (placenta extract, lactic acid fermentation metabolite). It was found that the activity was high.
[0016]
Next, the following test was conducted using the cream prepared in Example 1 above.
(Use test)
The face of 6 women was divided into left and right, and the cream of the example was used on one side and the cream of the comparative example was used once or more daily on the other side. After 3 months, rough skin prevention, skin gloss and skin Evaluation was made on the paste. In addition, a comparative example replaces the manufacture example with the purified water from the Example (comparative example).
[0017]
The evaluation is based on the following evaluation criteria, and the results are summarized in Table 2 below.
(Evaluation criteria)
The example is much better 3
The example is much better 2
Example is slightly better 1
No difference 0
Comparative example is slightly better -1
The comparative example is much better -2
The comparative example is much better -3
[0018]
[Table 2]
[0019]
As is clear from the above-mentioned use test (Table 2), the external preparation for skin (cream) of the present invention is effective for prevention of rough skin, skin gloss and skin elasticity, high safety, and an effect of enhancing skin activity. It was found to have
[0020]
【The invention's effect】
In the production method of the present invention, a skin external preparation material having a cell activation effect is obtained by subjecting rice bran and soybean peptide obtained from natural products as starting materials to fermentation with Bacillus subtilis, which is easily available, and then purifying it. Since it will be manufactured, it can manufacture safely and easily, without requiring a large sized installation.
The cell-utilizing skin external preparation of the present invention is very useful because it is highly safe and can enhance the activity of the skin.
Claims (3)
Priority Applications (1)
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JP24316994A JP3625501B2 (en) | 1994-10-06 | 1994-10-06 | Method for producing raw material for skin-applied skin and cell-applied skin external agent |
Applications Claiming Priority (1)
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JP24316994A JP3625501B2 (en) | 1994-10-06 | 1994-10-06 | Method for producing raw material for skin-applied skin and cell-applied skin external agent |
Publications (2)
Publication Number | Publication Date |
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JPH08104647A JPH08104647A (en) | 1996-04-23 |
JP3625501B2 true JP3625501B2 (en) | 2005-03-02 |
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JP24316994A Expired - Lifetime JP3625501B2 (en) | 1994-10-06 | 1994-10-06 | Method for producing raw material for skin-applied skin and cell-applied skin external agent |
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Families Citing this family (11)
Publication number | Priority date | Publication date | Assignee | Title |
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JP3240103B2 (en) * | 1995-04-28 | 2001-12-17 | 日鉱商事株式会社 | Detergent and method of using it |
EP1066816B1 (en) * | 1998-03-31 | 2007-08-01 | Shiseido Company Limited | Agents promoting laminin production in skin cells |
JP2002128683A (en) * | 2000-10-24 | 2002-05-09 | Umano Tekko Kk | Skin liniment or cleanser |
JP4612180B2 (en) * | 2000-12-19 | 2011-01-12 | 株式会社ヤクルト本社 | Skin preparation |
WO2002049656A1 (en) * | 2000-12-19 | 2002-06-27 | Kabushiki Kaisha Yakult Honsha | External skin preparations and process for producing the same |
JP4693623B2 (en) * | 2005-03-07 | 2011-06-01 | 共栄化学工業株式会社 | Cosmetics |
JP4945556B2 (en) * | 2006-05-02 | 2012-06-06 | 株式会社東洋発酵 | Cell application skin external preparation composition |
WO2008069102A1 (en) * | 2006-12-06 | 2008-06-12 | Calpis Co., Ltd. | Prophylactic/therapeutic agent for inflammatory bowel disease |
FR2918570B1 (en) * | 2007-07-09 | 2012-10-05 | Engelhard Lyon | DIGLYCATION OF AGEs. |
TWI393780B (en) * | 2007-08-09 | 2013-04-21 | Toyo Hakko Co Ltd | A fermentation composition, a cosmetic composition, and the like |
JP6332941B2 (en) * | 2013-11-01 | 2018-05-30 | 株式会社東洋発酵 | Oral composition for beauty health |
-
1994
- 1994-10-06 JP JP24316994A patent/JP3625501B2/en not_active Expired - Lifetime
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JPH08104647A (en) | 1996-04-23 |
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