JPS6327413A - Skin cosmetic - Google Patents

Abstract

PURPOSE:A safe and physico-chemically stable skin cosmetic, containing a cultivation filtrate of a fungus belonging to the genus Endomyces and having improved humectant effect. CONSTITUTION:A skin cosmetic containing a cultivation filtrate of a fungus, preferably Endomyces magnusii belonging to the genus Endomyces. The above- mentioned cosmetic, having extremely high safety without any possibility of allergy can be obtained by removing high polymers having >=13,000 molecular weight from the cultivation filtrate by ultrafiltration and filtering the resultant ultrafiltrate. The above-mentioned cultivation germfree filtrate or concentrate thereof may be directly used, but a preservative, etc., is preferably added to enhance the safety thereof. The cosmetic has improved humectant property and is capable of preventing the skin roughening and keratinization of the skin by keeping the moisture of the keratinous layer part of the skin at a normal value and sustaining the flexibility of the skin.

Description

Detailed Description of the Invention (Field of Industrial Application) The present invention is a new skin cosmetic product effective for moisturizing the skin (including medicinal cosmetics as quasi-drugs and external skin preparations as pharmaceuticals; the same applies hereinafter). In particular, the present invention relates to a skin cosmetic containing a culture filtrate of a bacterium belonging to the genus Endomyces as an active ingredient. More specifically, the present invention improves the moisture content of the stratum corneum of the skin through the comprehensive action of various amino acids, peptides, proteins, sugars, organic acids, inorganic salts, etc. contained in the culture filtrate of bacteria belonging to the genus Endomyces. The present invention relates to microbial preparations that are effective in preventing skin roughness, skin keratinization, etc., and maintaining skin flexibility by maintaining the skin in a normal state.

(Prior Art) In recent years, importance has been placed on the so-called usefulness of cosmetics, such as maintaining or improving skin health, and research to obtain technical support for this has been actively conducted.

In particular, the moisturizing mechanism of the stratum corneum of the skin plays an extremely important role in maintaining or improving skin health.

Normally, the stratum corneum of healthy skin contains 10 to 30% water, which maintains moisture and flexibility, but when it becomes less than 10% due to various reasons, the skin becomes dry and loses its flexibility and elasticity. This leads to loss of sexiness, which in turn leads to skin problems such as rough skin, rough skin, cracks, red spots, dandruff, and itching.

Furthermore, it is said that this can develop into age spots, fine wrinkles, etc. - Natural moisturizing factors (Natural moisturizing factors) are needed to moisturize living skin.
al Moisturizing Factor+
Hydrophilic components with high hygroscopicity and moisturizing properties such as amino acids (N, M, F,), pyrrolidone carboxylic acid, lactate, urea, and inorganic salts prevent their leakage and control the movement of moisture. It is known that the lipid components in the body play an important role.

Glycerin, propylene glycol, dipropylene glycol, etc. have traditionally been used in cosmetics for the purpose of moisturizing the skin.
．． 3-butylene glycol, polyethylene glycol,
Moisturizing agents such as polyhydric alcohol, pyrrolidone carboxylic acid, sugars, and amino acids are often used. Furthermore, in recent years, various efforts have been made to develop formulation systems that more closely approximate natural moisturizing factors; however, there are many unknown components of natural moisturizing factors, and their constituent components are complex and diverse, and their composition ratios are unclear. For these reasons, nothing truly effective could be obtained. Furthermore, in formulation, it is not only necessary to mix these components individually, but also there are various problems in maintaining the stability of the product, such as the formation of precipitates if the formulation is too complicated.

(Problems to be Solved by the Invention) An object of the present invention is to provide a novel material that is desirable, physicochemically stable, and safe as a cosmetic having an excellent moisturizing effect.

(Means for Solving the Problems) The present inventors searched widely for substances that can be used as cosmetics in nature, and particularly focused on microorganisms. As a result of a detailed study of various types of microorganisms and their culture conditions, we found that the culture filtrate of bacteria belonging to the genus Endomyces satisfies all the conditions required for a cosmetic product with excellent moisturizing effects. They discovered this and completed the present invention.

The bacteria belonging to the genus Endomyces used for the culture of the present invention are Endomyces magnusii (Endomyces magnus).
nusii), Endomyces geotrichum (End
o-myces geotrichum), Endomyces vessii
) etc. may be used, as long as the culture filtrate has an excellent moisturizing effect and has preventive and therapeutic effects on rough skin, skin keratosis, etc. Not particularly limited.

From the viewpoint of not causing cloudiness in the culture filtrate and having aromatic properties,
More preferably, Endomyces magnusii may be used. The strain used is, for example, Endomyces magnusii IFO4600, which is a known microorganism.
S. magnusii IFO4600) can be used after receiving distribution, but there is no need to be particularly limited, and those isolated from the natural world and artificial or natural mutants thereof can be used.

The method for culturing the bacteria belonging to the genus Endomyces is not particularly limited, and any conditions may be used as long as the bacteria used in the practice of this invention can grow well. The nitrogen source to be used may be anything as long as it can be assimilated by the bacteria belonging to the genus Endomyces. For example, organic nitrogen sources such as polypeptone, skim milk, yeast extract, malt extract, and amino acids may be used alone or in combination. can be used. The carbon source may be anything as long as it can be assimilated by the bacteria belonging to the genus Endomyces used, such as glucose, galactose, shakerose, etc.
Glycerin etc. can be used. If necessary depending on the medium, inorganic salts such as magnesium salts, sodium salts, and potassium salts, vitamins, amino acids, or substances rich in these may be added. The concentration of the above-mentioned assimilation substances varies depending on the type, but it is 0.1 g/j! ~100
g/2, and the culture temperature may be in the range of 20°C to 35°C. The pIi of the medium during culture is in the range of 3.5 to 8.5.

Cultivation is carried out by methods such as static culture, shaking culture, or aerated agitation culture. Although it varies depending on the culture conditions, it can be obtained by sterilizing and filtering after culturing for 2 to 7 days. At this time, ultrafiltration was performed using Ultrafiltration to remove polymeric substances with a molecular M of 13,000 or more from the culture filtrate, and the possibility of allergies was eliminated by filtration with a membrane filter. Cosmetics with extremely high safety can be obtained. The ultrafiltration membrane used here may be of any type as long as it can fractionate substances with a molecular weight of 13,000 or more. When using the cultured sterile filtrate thus obtained as a cosmetic, the cultured sterile filtrate or its concentrate can be used as is, but it is desirable to add a preservative or the like to increase its safety. It can also be used by blending it into ordinary lotions, emulsions, creams, etc. The cultured sterile filtrate thus obtained contains various amino acids, organic acids, pepquids, soluble proteins, alcohol, etc., and is particularly characterized by the fact that it produces a fruit ester aroma similar to apple and pineapple. That's true.

Next, an example of analysis of a sterile cultured filtrate of bacteria belonging to the genus Endomyces will be shown. The exemplified culture filtrate is (A
), (B), (C) and (D).

First, general analytical values are shown in Table 1.

(Left space below) Table 1 Next, qualitative and quantitative component analysis results are shown below for amino acids, organic acids, and soluble proteins that are considered to be closely related to the moisturizing effect of the culture filtrate.

As shown in the second amino acid table, it can be seen that there are many types of amino acids, which are one of the natural moisturizing factor components required for moisturizing the skin.

Organic Acids - As shown in the figure, citric acid and lactic acid, which are considered to be components of natural moisturizing factors, have been confirmed, but in addition to these, acetic acid, fumaric acid, lactic acid, isobutyric acid, and isovaleric acid are also found. and peaks of various unknown organic acids are observed. - Soluble protein (peptide) The molecular weight distribution of soluble protein is shown in Figure 2.

Although it is naturally predicted that these soluble proteins may be involved in the skin moisturizing mechanism, the possibility that they have some kind of physiologically active action including enzymatic action cannot be denied. However, this point cannot be clarified yet.

In addition, regarding the fruit ester aroma possessed by the culture filtrate, its components include isobutyl acetate, ethyl acetate, and
-Methylpropionic acid, 2- and 3-methylbutyric acid, etc. have been confirmed.

(Examples and Experimental Examples) Next, the present invention will be described in detail based on Examples and Experimental Examples.

Example 1 (Preparation of sterile culture filtrate) (A) A medium containing 3 g of glucose/5 g of 11 skim milk/β and 0.5 g of yeast extract/β was adjusted to pH 6.0 and then heat sterilized at 115° C. for 30 minutes. , Endomyces magnusii IFO4600 was inoculated and statically cultured at 25°C for 4 days. After sterilizing and filtering this culture, ultrafiltration was performed using Ultrafiltration, and the molecular weight was 13.
After removing 000 or more polymeric substances, a sterile filtrate (A) was obtained by filtration with a membrane filter.

(B) As a seed fungus, Endomyces isolated from nature
Cultivation was carried out in the same manner as in (A) using M. magnusi to obtain a sterile culture filtrate (B).

(C) Endomyces geotrichum IFO as the inoculum
Using 9541, culture was carried out in the same manner as in (A) above to obtain a culture sterile filtrate (C).

(D) As a seed fungus, Endomyces beige-IPO11
12 and cultured in the same manner as in (A) above,
A culture sterile filtrate (D) was obtained.

Experiment regarding moisturizing effect (A), (B), CC'') and (D) of Example 1 above
The cultured sterile filtrate of various bacteria belonging to the genus Endomyces obtained in ，
445 (1980)) are shown in Figure 3. However, as controls, physiological saline and a 2% aqueous solution of urea, which is often used as a moisturizing agent in cosmetics, were used. It can be seen that the culture sterile filtrate of the present invention has an excellent water retention ability.

Example 2 (Lotion) Prescription Weight % Cultured sterile filtrate (A) 90.0 Ethanol 5.0 Preservative
0.2 oval ice cube
The effect of the remaining lotion on rough skin was tested as follows.

(1) Subjects' skin roughness: 7 boys (28
23 women (22 to 56 years old) with smooth skin
: 3 boys (21-27 years old) and 9 girls with dry skin symptoms
(11 to 32 years old) Akagiri: 8 boys (7 to 29 years old) and 7 women (18 to 33 years old) with symptoms of red rash (2) Test method Subjects washed their faces with warm water twice a day, morning and evening. Afterward, the subjects were asked to apply a lotion, and 14 days later, the improvement effect on the symptoms was evaluated.

(3) Evaluation Each symptom was classified as follows according to its severity.

Skin roughness 1: The surface of the skin is dry, the edges tend to peel off, and the skin is constantly covered with white powder, and when scratched with the tip of the toe, the edges easily peel off along the lines.

2: The skin surface is dry, the corners are peeled off and a slight white powder is observed.

3: The skin surface is dry and rough, but the edges do not peel off and give off white powder.

4: The skin surface is moist and smooth.

Shark skin 1: The skin surface is dry, and clear fissures in the shape of fish scales can be seen all over the skin.

2: The skin surface is dry, and fish scale-like fissures can be seen all over the skin, but they are not very clear.

3: The skin surface is slightly dry, and partially indistinct fish scale-like fissures are visible.

4: There is moisture on the skin surface, and no fish scale-like fissures are observed in the eyes.

Akagiri ゛1: The skin surface is dry, and there are many incisions and spiral fissures.

2: The skin surface is dry, and spiral cracks are observed in some places.

3: The skin surface is slightly dry, and small spiral sassafres are slightly observed.

4: The skin surface is moist and no sassafre-like scars are observed.

The symptoms were evaluated as follows according to the classification of symptoms.

Effective: Symptoms improved from 1 to 3.4 and from 2 to 4.

Moderately effective: Symptoms 1 to 2, 2 to 3, and 3 to 4
improved on.

Ineffective: Symptoms have not changed significantly or have worsened.

The evaluation results are shown in Table 3.

(Margin below) Table 3 Example 3 (Cream) Formula Weight % Stearic acid 2.0 Stearyl alcohol 7.0 Squalane
5.0 reduced lanolin
2.0 octyldodecanol 6
．． 0 Polyoxyethylene cetyl ether (25E, O,) 3.0 Lipophilic glycerin monostearate 2.0 Preservative
0.3 antioxidant,
0.1 Culture sterile filtrate (A) 60.0 Purified water Remaining part The effect of the obtained cream on rough skin etc. was tested in the same manner as in Example 3. However, the number of subjects is as follows.

Skin irritation: 3 boys (36-42 years old), 7 girls (23 years old)
(Ages 52 to 52) Same skin: 10 females (16 to 39 years old) Dark skin: 4 males (19 to 33 years old), 6 females (32 to 57 years old) The evaluation results are shown in Table 4.

(Left below) Table 4 Example 4 (Hair Tonic) Prescription Weight % Sterile culture filtrate (A) 60.0 Ethanol 27.0 Preservative
0.1 Purified water Remaining part The effect of the obtained hair tonic on dandruff and itching was tested as follows.

(1) Subjects: 20 boys (22 to 43 years old) complaining of severe dandruff and itching.
(2) Test method Subjects were asked to apply hair tonic twice a day in the morning and evening.
After 14 days, the subjects were asked to self-evaluate the improvement effect.

(3) Evaluation Self-reported effects were classified as follows.

Effective: Dandruff and itching are almost completely gone.

Slightly effective: There is still some dandruff and itching, but it worked well.

Ineffective: There was little or no effect on dandruff and itching.

The evaluation results are shown in Table 5.

(The following is a blank space) Table 5 Furthermore, as shown in Table 6, the culture sterile filtrate of the present invention has long-term stability.

(The following is a blank space) Table 6 Furthermore, the safety test results of the culture sterile filtrate of the present invention are shown below.

Mucosal irritation test According to the Draize method, nine female white rabbits weighing 3.0 to 3.5 kg were divided into three groups of three rabbits each, and cultured sterile filtrate (
A) 0.1- was applied to the conjunctival sac of one eye of each rabbit, and the other eye served as a control. After the application, the - group was left untreated, the other group was washed with lukewarm water 2 seconds after the application, and the other group was washed with lukewarm water 4 seconds after the application. Observations were made. There was no irritation to the cornea, cornea, or conjunctiva.

A 10-fold concentrate of the sterile culture filtrate (A), a 10-fold concentrate of the sterile culture filtrate (B), and a 10-fold concentrate thereof were tested in the same manner, but no irritation was observed.

Skin - secondary irritation test 8 white rabbits weighing 2.0 to 2.51 qr were tested in groups of 4 each.
After dividing the animals into groups and cutting the hair on their backs, the - group left them as they were, and the other group made a scratch on the test area, and applied the culture sterile filtrate (A) on a 2.50 x 2.5 cIIl lint cloth. Apply to the skin. After 24 hours, the lint cloth was removed and observed, and then again after 48 hours and 72 hours.

No erythema, skin formation, or edema was observed in any of the groups.

In the same manner, tests were conducted on a 10-fold concentrated solution of the sterile culture filtrate (A), a 10-fold concentrated solution of the sterile culture filtrate (B), and no irritation was observed.

Skin sensitization test Out of 20 female guinea pigs weighing 320-350 g, 10 were used for sensitization treatment, and the other 10 were used as controls during induction.
An imization test was conducted. Using the culture sterile filtrate (A) and adjuvand, sensitization was performed by skin injection and then by application, and a provocation test was performed, and no changes were observed macroscopically.

In the same manner, tests were conducted on a 10-fold concentrated solution of the sterile culture filtrate (A), a 10-fold concentrated solution of the sterile culture filtrate (B), and no sensitization was observed.

In addition, when a test was conducted using the same method on the liquid from the ultrafiltration section of Ultrafiltration in Example 1 (A), one out of 10 mice showed slight erythema for the 10 times concentrated liquid. However, it was not a problem.

Human Patch Test A closed patch test was conducted on the culture sterile filtrate (A), the culture sterile filtrate (B), and their respective 10-fold concentrates. AI-test (IMECOAstra＾
Place each sample 0 on the filter paper section of the Gency Co.
．． A closed patch was applied to the inside of the upper arm of 40 men for 48 hours, and the results were evaluated at 1 and 24.96 hours, but none of them showed a positive result.

(Effect of the invention) As is clear from the component analysis, examples, and experimental examples above,
The culture filtrate of the present invention contains amino acids, organic acids, soluble proteins, and many other hygroscopic and moisturizing substances such as organic substances, inorganic salts, and alcohol, whether they are known substances or unknown substances. Regardless of the ingredients, it is a stable and safe makeup that has excellent efficacy that cannot be obtained with conventional cosmetics that are formulated with individual moisturizers because it is efficiently contained in an appropriate balance. It is a fee. Furthermore, it has excellent effects on the prevention and treatment of various skin troubles that occur as a result of water imbalance, especially rough skin, flaky skin, cracks, red spots, dandruff, itching, and the like. Furthermore, the fruit ester aroma of the main culture filtrate provides excellent performance for cosmetics that rely on fragrance, and does not require the addition of fragrances, which are considered problematic from a safety perspective.
This product has the above advantages and provides a more excellent cosmetic product.

[Brief explanation of the drawing]

Figure 1 shows that organic acids are components of natural moisturizing factors, Figure 2 shows the molecular weight distribution of soluble proteins, and Figure 3 shows the molecular weight distribution of various bacteria belonging to the genus Endomyces. FIG. 3 is a diagram showing the water retention ability of a sterile culture solution. Patent applicant Sugiura New Drug Development Institute Co., Ltd.

Claims (1)

[Scope of Claims] 1. A skin cosmetic containing a culture filtrate of a bacterium belonging to the genus Endomyces. 2. The skin cosmetic according to claim 1, wherein the bacterium belonging to the genus Endomyces is Endomyces magnusii. 3. The skin cosmetic according to claim 1, wherein the culture filtrate is a sterile filtrate from which polymeric substances having a molecular weight of 13,000 or more have been substantially removed. 4. The skin cosmetic according to claim 2, wherein the polymer substance is removed by an ultrafiltration membrane.