JP3103747B2 - Cosmetics - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a novel cosmetic containing a culture having an excellent lipid peroxide production inhibitory effect and a moisturizing effect. More specifically, Zymomonas bacteria ( Zym
Omonas mobilis ), comprising a culture obtained by liquid culturing of Omonas mobilis ) and a lipid peroxide production inhibitor.

[0002]

2. Description of the Related Art In recent years, it has been reported that the production and accumulation of lipid peroxide generated in the skin due to exposure to ultraviolet rays, etc., has affected skin aging (wrinkles, sagging, senile pigments). Substances that suppress the production of lipid peroxides in the skin have been attracting attention for the purpose of prevention.

Conventionally, radical scavengers such as vitamin E, vitamin C, vitamin A, glutathione and uric acid have been known as inhibitors of lipid peroxide in the skin, but they directly inhibit lipid peroxidation in cells. What you do
Only vitamin E is used. Although this vitamin E exhibits an excellent lipid peroxide-suppressing effect in vivo, there is a problem that its application range is limited because it is fat-soluble and hardly soluble in water.

[0004] The moisturizing mechanism of the stratum corneum of the skin plays an extremely important role in maintaining or improving the health of the skin. The stratum corneum of healthy skin usually contains 10 to 30% of water and maintains moisture and softness. However, when the water content is 10% or less due to various causes, the skin becomes dry and soft. Loss of elasticity, and eventually rough skin,
Skin troubles such as shark skin, cracks, red ash, dandruff, and Kayumi tend to occur. In addition, it is thought that it progresses to spots, fine lines, and the like. For moisturizing the skin, natural moisturizing factors (Natural Moisturizing)
Factor = N. M. F. ) Is known to play an important role. This N. M. F. Consists of hydrophilic components such as amino acids, pyrrolidone carboxylic acid, lactic acid, urea, and inorganic salts, which are highly hygroscopic and moisturizing, and lipid components that prevent their outflow and control the movement of water. [H.
W. Spier et al., Hautartzt, Volume 7, Page 2 (1956)].

In conventional cosmetics, moisturizing agents such as glycerin, propylene glycol, dipropylene glycol, 1,3-butylene glycol, polyethylene glycol, polyhydric alcohol, pyrrolidone carboxylic acid, saccharides and amino acids have been used for the purpose of moisturizing the skin. Is blended. Further, in recent years, various devices have been devised to develop a prescription system that approaches a natural moisturizing factor. However, because there are many unknown parts in the natural moisturizing factor, and its components are complex and diverse and the composition ratio is not exactly known, the same formulation as the natural moisturizing factor has not been developed . Also, there are various problems in maintaining the stability of the product, for example, when a substance close to these is synthesized and mixed, a precipitate is generated.

[0006] There is no application of a culture of Zymomonas bacteria to cosmetics, and no effect as a lipid peroxide inhibitor is known.

[0007]

DISCLOSURE OF THE INVENTION In view of the above-mentioned situation, the present inventors have an excellent lipid peroxide production inhibitory effect and a moisturizing effect, and are physicochemically stable and have no problem in safety. With the aim of providing a novel cosmetic, the effectiveness of a culture of Zymomonas bacteria was examined, and it showed an excellent inhibitory effect on lipid peroxide production, and characterized by containing a culture of Zymomonas bacteria. The present invention has been completed because cosmetics exhibit excellent moisturizing effects and are stable over time when formulated.

[0008]

That is, the present invention is a skin cosmetic comprising a liquid culture of Zymomonas bacteria, and a culture obtained by the culture.

The Zymomonas bacteria used in the present invention ( Zy
momonas mobilis ) is an alcohol-fermenting bacterium belonging to the Gram-negative bacterium.
European spoiled beer, fermented apple juice (ci
der) and pear juice (perry), and other tropical p
It is also isolated from alm wines and fermented sugarcane juice in Brazil. Also in the ripening honey. Zymomonas bacteria are widely available in the wild and easily available, or can be obtained from, for example, the Fermentation Research Institute (Foundation, Yodogawa-ku, Osaka) or the American Type Culture Collection (ATCC) (IFO 13756, IFO13757, ATCC).
31821, ATCC 31822, ATCC 31823, ATCC 35000, ATCC 35
001, ATCC 10988, ATCC 29191, ATCC 29192).

In the present invention, glucose, sucrose, fructose or a mixture thereof is used as a carbon source component of a medium for cultivating Zymomonas bacteria (2 based on the volume of the medium).
２０20 w / v%). Examples of the nitrogen source component that can be used in the medium include amino acids, peptides, ammonium salts, nitrates, peptone, skim milk, and mixtures thereof. In addition, as inorganic salt components that can be used in the medium, sodium chloride, magnesium sulfate, phosphate,
Examples thereof include ferrous sulfate, potassium chloride, and mixtures thereof. As other medium components, nutrient components such as malt extract, potato extract, yeast extract, oatmeal, and wheat malt may be added. The culturing conditions are preferably a temperature of 20 to 37 ° C. and a pH of 2.5 to 7.0, and the culturing is preferably performed for 1 to 4 days by a method such as stationary culturing, shaking culturing, or aeration culturing. Although the main culture can be carried out from the beginning, it is preferable to carry out a small-scale preculture in advance and inoculate the main culture with the preculture.

After the liquid culture, the culture extract containing the culture components secreted outside the cells by the Zymomonas cells obtained by removing the cells of the Zymomonas cells by a conventional method is used as a cosmetic product. Can be used as raw material. In this eradication extraction, for example, the culture solution after liquid culture is 1000-5000.
It can be carried out by centrifugation giving a gravitational acceleration of g, or by filtration with a filter medium such as Hyflo supercell or Celite. Further, the filtrate thus obtained is further fractionated and purified by a method such as dialysis, removal of low molecular components by ultrafiltration, etc., deproteinization operation, fractionation using a solvent such as alcohol, and the like. You can also. In addition, the culture components after the removal of the cells can be further fractionated and the cultured components can be concentrated or diluted as necessary. Further, it can be used as a cosmetic raw material in a dry powder state by a method such as freeze drying or spray drying.

[0012] Accordingly, as used herein, "culture"
The term is a culture extract from which cells have been removed after liquid culture,
It means a purified product of a culture extract, a concentrated or diluted product thereof, or a dried product thereof.

The culture thus obtained was found to have a remarkable effect of suppressing lipid peroxide production.

[0014] The cosmetic containing the culture is
It was possible to improve the color of the skin and leave a good protective film with a very light feeling upon use. Further, the cosmetic is
It was found to have very significant moisturizing properties.

In the cosmetic of the present invention, raw materials such as oils and fats, waxes, hydrocarbons, fatty acids, alcohols, esters, and surfactants used for cosmetics and medicines are used for the culture of Zymomonas bacterium. It can be blended within a range that does not impair the effect. The cosmetics of the present invention include creams, lotions,
Base cosmetics such as emulsions and packs, makeup cosmetics such as foundations and lipsticks, bath preparations, soaps and other forms can be employed.

The amount of the Zymomonas bacterium culture in various cosmetics is not particularly limited since it can be varied depending on the form. In principle, it is sufficient that an effective amount is present, but it is generally preferable to add 0.0001 to 99% of the cosmetic composition (total weight).

The culture of Zymomonas bacteria has no toxicity and irritation to the skin (according to the cumulative irritation test and the patch test method) and is highly safe against light and heat.

[0018]

EXAMPLES Next, the present invention will be described in detail with reference to Examples, but the present invention is not limited thereto. The parts of the amounts shown in the examples are parts by weight.

[Preparation method of medium] Medium of a) Dextrose 50 g Potassium dihydrogen phosphate 2 g Magnesium sulfate 0.5 g Yeast extract 1 g Soybean peptide 10 g Distilled water 1000 ml NaOH or HCl adjusted to pH 6.0 I do.

Medium b) Sucrose 50 g Potassium dihydrogen phosphate 2 g Yeast extract 10 g Polypeptone 10 g Distilled water 1000 ml Adjust the pH to 6.0 with NaOH or HCl.

Medium c) Fructose 50 g Potassium dihydrogen phosphate 2 g Magnesium sulfate 0.5 g Ammonium sulfate 5 g Malt extract 10 g Potato extract 10 g Distilled water 1000 ml Adjust the pH to 6.0 with NaOH or HCl.

A liquid medium was prepared by subjecting the medium having the compositions a) to c) to high-pressure steam sterilization (120 ° C., 20 minutes).

[Culture Method of Zymomonas Bacteria] Zymomonas bacteria (IFO 13756 strain) were cultured in the liquid medium of the above a) to c) for 3 days, pre-cultured in advance, and the culture solution obtained by the pre-culture was separately added. One hundredth of the liquid medium described in a) to c) above is inoculated corresponding to the medium used in the preculture, and cultured by static culture at a temperature of 30 ° C and a pH of 6.0 for 4 days.

[Method of Extracting Culture Solution] After the liquid culture, the culture solution is prepared by removing the cells by a conventional method, and using the culture extract containing the culture components secreted out of the cells as a cosmetic raw material. can do. This eradication extraction can be performed, for example, by subjecting the culture solution after liquid culture to centrifugation at a gravitational acceleration of 1000 to 5000 g.
In the following Examples, centrifugal separation was performed by applying a gravitational acceleration of 5000 ± 200 g, and a liquid obtained by removing cells of Zymomonas bacteria was used as a cosmetic raw material as it was.

Using the above liquid culture media a) to c),
The culture extracts obtained as described above were each extracted with extract a)
To extract c).

Example-1 Lotion Formulation Formulation amount 1. Ethanol 5.0 parts 2. Methyl P-oxybenzoate 0.1 3. Polyoxyethylene (40) hydrogenated castor oil 0.1 4. Perfume proper amount 5. Purified water 10.0 6. Zymomonas Bacterial liquid culture extract b) 5.0 7.1,3-butylene glycol 3.0 8. Glycerin 2.0 9. Xanthan gum 0.02 10. Purified water 69.6 Production method: Components 1 to 5 and components 6 to 10 are uniformly dissolved and mixed, respectively. Filter to make the product.

Comparative Example 1 Conventional lotion In Example 1, a culture extract of Zymomonas bacteria b)
Was replaced with purified water to obtain a conventional lotion.

Example-2 Cream Formulation Formulation amount 1. Liquid paraffin 6.5 parts 2. Vaseline 10.0 3. Stearic acid 4.0 4. Cetyl alcohol 3.0 5. Stearyl alcohol 1.0 6. Polyoxyethylene (25) monostearate 3.0 7. Sorbitan monostearate 2.5 8.1,3-butylene glycol 5.0 9. Potassium hydroxide 0.1 10. Liquid culture extract of Zymomonas bacteria a) 1.0 11. Methyl P-oxybenzoate 0.2 12. Purified water 64.5 13. Perfume Appropriate amount Production method : Oil phase components 1 to 7 and water phase components 8 to 12 are heated and dissolved at 70 to 75 ° C respectively, and then oil phase components 1 to 7 are converted to aqueous phase components.
Add 8 to 12 to emulsify, add and mix component 13 while cooling, and cool to 30 ° C to obtain a product.

Example 3 Emulsion Formulation Formulation amount 1. Liquid paraffin 8.0 parts 2. Squalane 2.0 3. Stearyl alcohol 2.0 4. Sorbitan monostearate 2.5 5. Glycerin monostearate 2.3 6. Polyoxyethylene (10) sorbitan monooleate 0.8 7. Glycerin 6.0 8. Liquid culture extract of Zymomonas bacteria b) 10.0 9. Methyl P-oxybenzoate 0.2 10. Purified water 67.0 11. Appropriate amount of fragrance Production method: Oil phase components 1 to 6 and aqueous phase components 7 to After heating and dissolving 10 to 70-75 ° C respectively, the aqueous phase components 1 to 6 are combined with the aqueous phase components.
Add 7 to 10 to emulsify, add component 11 while mixing, mix and cool to 30 ° C to obtain a product.

Comparative Example 2 Conventional Emulsion In Example 3, culture extract of Zymomonas bacteria b)
Was replaced with purified water to obtain a conventional emulsion.

Example-4 Pack Formulation Formulation Amount 1. Polyvinyl alcohol 12.0 parts 2. Ethyl alcohol 5.0 3.1, 3-butylene glycol 8.0 4. Methyl P-oxybenzoate 0.2 5. Polyoxyethylene (40) hydrogenated castor oil 0.5 6. Liquid culture extract of Zymomonas bacteria c) 2.0 7. Citric acid 0.1 8. Sodium citrate 0.3 9. Fragrance 0.1 10. Purified water 72.8 Production method: Each component is uniformly dissolved to produce a product.

Example-5 Foundation Formulation Compounding amount 1. 2.4 parts of stearic acid 2. Polyoxyethylene (20) sorbitan monostearate 1.0 3. Polyoxyethylene (20) cetyl ether 2.0 4. Cetyl alcohol 1.0 5. Liquid lanolin 2.0 6. Liquid paraffin 3.0 7. Isopropyl myristate 6.5 8. Butyl oxybenzoate 0.1 9. Purified water 53.2 10. Liquid culture extract of Zymomonas bacteria a) 5.0 11. Sodium carboxymethylcellulose 0.1 12. Bentonite 0.5 13. Propylene glycol 4.0 14. Triethanolamine 1.1 15. Methyl P-oxybenzoate 0.2 16. Titanium oxide 8.0 17. Talc 4.0 18. Color pigment 5.0 19. Perfume qs Production method: Components 11 to 13 and 15 heated to 70 ° C Swell well. Components 9 to 10 and 14 are dissolved in this to form an aqueous phase. Components 1 to 8 are dissolved by heating and kept at 80 ° C. to form an oil phase. The components 16 to 18 mixed well and passed through a pulverizer are added to the aqueous phase, and the mixture is stirred with a homomixer and kept at 75 ° C. Add the oil phase to the aqueous phase with stirring, cool, add component 19 at 45 ° C,
Cool with stirring to obtain a product.

Example -6 Hair cream Formulation Formulation amount 1. Stearic acid 2.0 parts 2. Stearyl alcohol 2.0 3. Liquid paraffin 8.0 4. Glycerin monostearate 2.3 5. Sorbitan monooleate 2.5 6. Polyoxyethylene (10) sorbitan Monooleate 0.8 7. Liquid culture extract of Zymomonas bacteria c) 0.5 8. Glycerin 6.0 9. Methyl P-oxybenzoate 0.1 10. Purified water 75.8 11. Appropriate amount of fragrance Production method: Oil phase components 1 to 6 and aqueous phase components Dissolve 7-10 at 70-75 ° C respectively, emulsify by adding aqueous phase component 7-10 to oil phase component 1-6, add component 11 while cooling, mix at 30 ° C
Cool down to a product.

[0034]

Industrial Applicability The culture of Zymomonas bacterium of the present invention has an excellent lipid peroxide production inhibitory effect, and the cosmetic containing the culture of Zymomonas bacterium has an excellent moisturizing effect. Next, experimental examples will be described in order to explain the effects of the present invention in detail. Experimental Example-1 Lipid peroxide production inhibition test The following method was used to examine the lipid peroxide production inhibition effect. 0.1 ml of a test substance is added to 0.9 ml of 10 mM linoleic acid / 0.1 M phosphate buffer (pH 7), and a medium-wavelength ultraviolet lamp (313 nm, manufactured by Toshiba, FL20S)
-E) was irradiated at 25 ° C. for 180 minutes. To 0.1 ml of the suspension irradiated with the medium wavelength ultraviolet ray, 8.1% SDS (sodium dodecyl sulfate) and 0.75 ml of 0.8% TBA were added.
(Thiobarbituric acid), 20% acetic acid (pH 3.5) and 0.3 ml of distilled water were added and reacted at 100 ° C. for 1 hour. This reaction solution was treated with 1-butanol / pyridine (1
5: 1), and the absorbance was measured at 535 nm.
The TBA ratio relative to the control (without addition of the test substance) was shown as a peroxidation inhibition rate. As a result, the culture of Zymomonas bacteria exhibited an excellent lipid peroxide production inhibitory action even at a low concentration (Table 1). (Below)

[0035]

[Table 1] Table 1 Lipid peroxide production inhibition test ──────────────────────────────────── Test substance Lipid peroxide production inhibition rate ──────────────────────────────────── Zymomonas liquid culture extract a) 100% aqueous solution 90% Zymomonas bacteria liquid culture extract a) 50% aqueous solution 90% Zymomonas bacteria liquid culture extract a) 25 aqueous solution 90% Zymomonas bacteria liquid culture extract b) 100% aqueous solution 100% Zymomonas bacteria liquid culture extract b B) 25% aqueous solution 100% zymomonas bacteria liquid culture extract c) 50% aqueous solution 95% zymomonas bacteria liquid culture extract c) 50% aqueous solution 95% zymomonas bacteria liquid culture extract c) 5% aqueous solution 90% α-tocopherol (25 mM) 100% blank (water) 0% ─────────────────────────────── ─────

Experimental Example 2 Moisturizing Effect Test The moisturizing effect of the above Zymomonas bacillus liquid culture extracts a) to c) was determined by measuring the hydration state of the stratum corneum in the living body [SKI
CON 200 (manufactured by IBS Inc., Hamamatsu City)]
Measured using An aqueous sodium hyaluronate solution recognized as having a moisturizing property in the art was used as a comparative example, and distilled water was used as a control example. The former device is 3.5MHz
The moisturizing effect of the stratum corneum of the skin is measured by measuring the electrical resistance of the stratum corneum of the skin at the following frequency.

The test was conducted on a total of 10 women and 5 men. An arbitrary 6 points (about 2 × 2 cm) inside the left and right forearms of each subject are divided (12 for each subject).
Part), each of Zymomonas bacteria liquid culture extract a)-
c), an area to which an aqueous solution of sodium hyaluronate (0.075% by weight) and distilled water were applied, and the remaining one area was an area to which nothing for blank test was applied.

The moisturizing state of each area before application was measured by the above hydration state measuring apparatus, and these measured values were defined as M ₀ .
Then, apply 500 μl of each sample to each area, and
Moisturizing state when passed 0 min was measured, these measurements was M _30. Table 2 shows the moisturizing effect values determined from the average values of the obtained M ₀ and M ₃₀ by the following formulas.

As can be seen from Table 2, the extracts a) to c) of the present invention showed a moisturizing effect comparable to or better than that of sodium hyaluronate.

Table 2 Moisturizing effect of the liquid culture extract of Zymomonas bacteria ─────────────────────────────────── Sample Moisturizing effect (%) ─────────────────────────────────── Zymomonas liquid culture extract a) 72 0.3 Zymomonas liquid culture extract b) 78.2 Zymomonas liquid culture extract c) 74.5 Sodium hyaluronate aqueous solution (comparative example) 68.5 distilled water (control example) 12.3 ─────────────────────────────

Experimental Example-3 Usage Test Using the lotion of Example-1, the emulsion of Example-3, the conventional lotion of Comparative Example-1 and the conventional emulsion of Comparative Example-2,
One month of use test was performed on 30 females (30 to 45 years old) each as a control. After use, the moisturizing effect was determined by a questionnaire survey on a moist feeling, a smooth feeling, a refreshing feeling and a sticky feeling. As a result, the cosmetics of the present invention showed remarkably superior moisturizing effects as compared with the comparative cosmetics (Tables 3, 4, 5, and 6).

[0041]

[Table 3] Table 3 Moisturizing action of lotion of the present invention (number of persons surveyed) ──────────────────────────────化粧 Lotion of the present invention (Example-1) Evaluation ────────────────────────── Very good Good Normal ─ ─────────────────────────────────── Moist 20 7 3 Smooth 20 8 2 Refresh 19 6 5 Non-stick 20 6 4 ──────────────────────────────────── (Margin below)

[0042]

[Table 4] Table 4 Conventional moisturizing effect of lotion (number of people surveyed) ─────────────────────────────── ───── Conventional lotion (Comparative Example-1) Evaluation ────────────────────────── Very good Good Normal ─── ───────────────────────────────── Moist 1 8 21 Smooth 2 9 19 Refresh 2 9 19 Non-sticky 1 7 22 ────────────────────────────────────

[0043]

[Table 5] Table 5 Moisturizing action of the emulsion of the present invention (number of persons in questionnaire results)乳 Emulsion of the present invention (Example-3) Evaluation ────────────────────────── Very good Good Normal ─── ───────────────────────────────── Moist 19 8 3 Smooth 16 11 3 Refreshing 22 5 3 Non-sticky 20 4 4 ──────────────────────────────────── (margin)

[0044]

[Table 6] Table 6 Moisturizing effect of conventional emulsion (number of people surveyed) 結果──── Conventional emulsion (Comparative Example-2) Evaluation ────────────────────────── Very good Good Normal ───── ─────────────────────────────── Moist 2 10 20 Smooth 4 8 18 Refresh 3 9 18 Non-sticky 4 10 16 ─────────────────────────────── ──────────────────────────────────

As described above, the culture of Zymomonas bacterium of the present invention is safe and exhibits an excellent lipid peroxide production inhibitory effect, and further contains a viscous substance which enhances the skin moisturizing effect. Therefore, the cosmetics containing the Zymomonas bacterium culture provide an extremely excellent cosmetic having an excellent effect of suppressing the production of lipid peroxide in the skin by the ultraviolet rays and a moisturizing effect, and the culture is industrially and stably produced. It has a remarkable effect of attaining the object of the invention.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification code FI A61P 39/06 A61P 39/06

Claims (2)

(57) [Claims] 1. A cosmetic comprising a culture of Zymomonas bacteria. 2. A lipid peroxide production inhibitor comprising a culture of Zymomonas bacteria.