JPH08337512A - Cosmetic - Google Patents

Abstract

PURPOSE: To provide a cosmetic and an inhibitor of peroxidized lipid formation containing the culture mixture of a microorganism in Zymomonas. CONSTITUTION: The objective cosmetic and inhibitor of peroxidized lipid formation contains a culture product of a microorganism in Zymomonas. A culture product is prepared, for example, by subjecting a microorganism in Zymomonas to liquid culture and separating its cell bodies from the culture mixture by centrifugation and the supernatant is utilized as an active ingredient. The objective cosmetic and the objective peroxide lipid formation inhibitor are provided in the preparation form of powder, pill, tablet, preparation for external use, injection solution, suppository and the like.

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 product having an excellent lipid peroxide production inhibitory effect and a moisturizing effect. More specifically, Zymomonas ( Zym
The present invention relates to a cosmetic and a lipid peroxide production inhibitor characterized by containing a culture obtained by liquid-culturing ( Omonas mobilis ).

[0002]

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

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

The moisturizing mechanism of the stratum corneum of the skin plays a very important role in maintaining or improving the health of the skin. The stratum corneum of healthy skin usually contains 10 to 30% of water, which maintains moisture and flexibility, but when the water content falls below 10% due to various causes, it becomes dry skin and softness. Or lack of elasticity, and eventually rough skin,
Skin problems such as shark skin, cracks, red fins, dandruff, and kayumi tend to occur. Furthermore, it is believed that it progresses to spots, fine lines, etc. Natural Moisturizing Factor (Natural Moisturizing)
Factor = N. M. F. It is known that the hygroscopic material called) plays an important role. This N. M. F. Is composed of hydrophilic components such as amino acids, pyrrolidonecarboxylic acid, lactic acid, urea, and inorganic salts, which have high hygroscopicity and moisture retention, and lipid components that prevent their outflow and control the movement of water. It has been reported [H.
W. Spier et al., Hautarzt, Vol. 7, page 2 (1956)].

Moisturizers such as glycerin, propylene glycol, dipropylene glycol, 1,3-butylene glycol, polyethylene glycol, polyhydric alcohol, pyrrolidonecarboxylic acid, sugars and amino acids for the purpose of moisturizing the skin in conventional cosmetics. Is included. Furthermore, in recent years, various efforts have been made to develop a prescription system that approaches a natural moisturizing factor. However, due to the fact that there are many unknown parts in natural moisturizing factors, their constituent components are complex and diversified, and the composition ratio is not clear exactly, a formulation equivalent to natural moisturizing factors has not been developed. . In addition, there are various problems in maintaining the stability of the product such as precipitation when synthesizing and blending those close to these.

[0006] There is no application example of a culture of Zymomonas bacteria to cosmetics, and its effect as a lipid peroxide inhibitor is not known.

[0007]

In view of the above situation, the present inventors have an excellent lipid peroxide production inhibitory effect and a moisturizing effect, are physicochemically stable, and have no safety problem. When the effectiveness of the culture of Zymomonas bacteria was examined for the purpose of providing a novel cosmetic, it showed an excellent inhibitory effect on lipid peroxide production, and was characterized by containing Zymomonas culture. The present invention has been completed because the cosmetics used have an excellent moisturizing effect and are stable over time when formulated.

[0008]

[Means for Solving the Problems] That is, the present invention is a skin cosmetic characterized in that a Zymomonas bacterium is liquid-cultured and the culture thus obtained is contained.

Zymomonas bacterium ( Zy used in the present invention
momonas mobilis ) is an alcohol-fermenting bacterium that belongs to Gram-negative bacteria, and is used in the sake called pulke made from the juice of Agave, a Longan orchid in Mexico,
European spoiled beer, fermented apple juice (ci
der) and pear juice (perry), and other p
It is also separated from alm wines and fermented sugarcane juice in Brazil. Also in the ripening honey. The Zymomonas bacterium is widespread and easily available in the wild.
31821, ATCC 31822, ATCC 31823, ATCC 35000, ATCC 35
001, ATCC 10988, ATCC 29191, ATCC 29192) are available.

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

[0011] The liquid culture after the liquid culture is a cosmetic extract containing a culture component secreted outside the cells of Zymomonas cells obtained by removing bacteria of Zymomonas cells by a conventional method. It can be used as a raw material. This sterilization extraction is carried out, for example, by adding 1000 to 5000 to the culture solution after liquid culture.
It can be carried out by centrifuging to give a gravitational acceleration of g or by filtering with a filter medium such as Hyflo Supercell or Celite. The filtrate thus obtained is further fractionated and purified by a method such as dialysis, removal of low-molecular components by ultrafiltration, deproteinization, fractionation using a solvent such as alcohol, and the like. You can also In addition, the culture solution obtained by further fractionating and purifying the culture broth after removal of the bacterial cells 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.

Accordingly, "culture" as used herein
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 showed a remarkable inhibitory effect on lipid peroxide production.

Further, the cosmetic containing the above-mentioned culture is
It was possible to improve the skin color and leave a good protective film with a very refreshing feel. Further, the cosmetic is
It was found to have very pronounced moisturizing properties.

In the cosmetic of the present invention, raw materials such as oils and fats, waxes, hydrocarbons, fatty acids, alcohols, esters, surfactants and the like used in cosmetics and medicines are prepared from cultures of Zymomonas bacteria. It can be blended within a range that does not impair the effect. The cosmetics of the present invention include creams, lotions,
A basic cosmetic such as an emulsion or a pack, a makeup cosmetic such as a foundation or a lipstick, a bath preparation, a dosage form such as soap can be adopted.

The amount of the Zymomonas culture cultivated in various cosmetics can be changed depending on its form, and is not particularly limited. In principle, it suffices if an effective amount is present, but generally 0.0001 to 99% is preferably blended with respect to the cosmetic composition (total weight).

Zymomonas cultures have no toxicity or irritation to the skin (according to the cumulative irritation test and the patch test method) and are highly safe against light and heat.

[0018]

EXAMPLES Next, examples will be given to explain the present invention in detail, but the present invention is not limited thereto. The parts of the compounding amounts shown in the examples are parts by weight.

[Preparation Method of Medium] Medium of a) Glucose 50 g Potassium dihydrogen phosphate 2 g Magnesium sulfate 0.5 g Yeast extract 1 g Soybean peptide 10 g Distilled water 1000 ml Adjust pH to 6.0 with NaOH or HCl. To do.

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

Medium of 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 to pH 6.0 with NaOH or HCl.

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

[Method for culturing Zymomonas bacterium] Zymomonas bacterium (IFO 13756 strain) is cultivated in the liquid medium of the above a) to c) for 3 days and precultured in advance, and the culture solution obtained by the preculture is The liquid medium of the above a) to c) is inoculated in an amount of 1/100 corresponding to the medium used in the pre-culture, and cultured by static culture at a temperature of 30 ° C., pH 6.0 for 4 days.

[Extraction Method of Culture Solution] After the liquid culture, the culture solution is removed from the cells by a usual method, and the culture extract containing the culture components secreted by the cells outside the cells is used as a cosmetic raw material as it is. can do. This sterilization extraction can be performed, for example, by centrifuging the culture solution after the liquid culture, which gives a gravitational acceleration of 1000 to 5000 g.
In the following examples, centrifugation was performed to give a gravitational acceleration of 5000 ± 200 g, and the liquid obtained by removing the bacteria of Zymomonas was used as the raw material for cosmetics.

Using the above liquid medium a) to medium c),
The culture extracts obtained as described above are respectively extracted a)
~ Extract c).

Example-1 Lotion Formulation Compounding 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 Liquid culture extract of fungus b) 5.0 7.1,3-butylene glycol 3.0 8. Glycerin 2.0 9. Xanthan gum 0.02 10. Purified water 69.6 Manufacturing method: Components 1 to 5 and components 6 to 10 are uniformly dissolved and mixed. The product is filtered.

Comparative Example-1 Conventional lotion In Example-1, culture extract of Zymomonas b)
What was replaced with purified water was the conventional lotion.

Example-2 Cream 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 a) 1.0 11. Methyl P-oxybenzoate 0.2 12. Purified water 64.5 13. Perfume Proper production method : Oil phase components 1 to 7 and aqueous phase components 8 to 12 are heated and dissolved at 70 to 75 ° C, respectively, and then oil phase components 1 to 7 are dissolved in water phase components.
Add 8 to 12 to emulsify, add component 13 during cooling, mix, and cool to 30 ° C to obtain a product.

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

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

Example-4 Pack 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 c) 2.0 7. Citric acid 0.1 8. Sodium citrate 0.3 9. Perfume 0.1 10. Purified water 72.8 Production method: Dissolve each component uniformly to obtain a product.

Example-5 Foundation Formulation amount 1. Stearic acid 2.4 parts 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 P-hydroxybenzoate 0.1 9. Purified water 53.2 10. Liquid culture extract of Zymomonas 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. Coloring pigment 5.0 19. Perfume suitable amount Manufacturing method: Ingredients 11 to 13 and 15 are heated to 70 ° C. Swell well. Ingredients 9 to 10 and 14 are dissolved in this to obtain an aqueous phase. Ingredients 1 to 8 are dissolved by heating and kept at 80 ° C to form an oil phase. Ingredients 16-18, which have been thoroughly mixed and passed through a grinder, are added to the aqueous phase, and the mixture is stirred with a homomixer and kept at 75 ° C. Add the oil phase to this water phase while stirring, cool and add component 19 at 45 ° C,
Cool with stirring to make the product.

Example-6 Hair Cream 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 c) 0.5 8. Glycerin 6.0 9. Methyl P-oxybenzoate 0.1 10. Purified water 75.8 11. Perfume Suitable amount Production method: Oil phase components 1 to 6 and water phase component Dissolve 7 to 10 at 70 to 75 ° C, respectively, add water phase components 7 to 10 to oil phase components 1 to 6 to emulsify, and add component 11 during cooling to mix, 30 ° C
Cool down to product.

[0034]

EFFECTS OF THE INVENTION The culture of Zymomonas bacterium of the present invention has an excellent inhibitory effect on lipid peroxide production, and the cosmetic containing the culture of Zymomonas bacterium has an excellent moisturizing effect. Next, in order to explain the effect of the present invention in detail, an experimental example will be given. Experimental Example-1 Lipid Peroxide Production Inhibition Test The following method was used to examine the lipid peroxide production inhibitory effect. 0.1 ml of the test substance was 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) was used.
-E) was irradiated at 25 ° C for 180 minutes. To 0.1 ml of the suspension irradiated with medium wavelength ultraviolet rays, 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 the mixture was reacted at 100 ° C. for 1 hour. This reaction solution was mixed with 1-butanol / pyridine (1
5: 1) and the absorbance was measured at 535 nm.
The TBA ratio with respect to the control (no addition of the test substance) was shown as the peroxide inhibition rate. As a result, the culture of Zymomonas showed an excellent lipid peroxide production inhibitory action even at low concentrations (Table 1). (Below margin)

[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 liquid culture extract a) 50% aqueous solution 90% Zymomonas liquid culture extract a) 25 aqueous solution 90% Zymomonas liquid culture extract b) 100% aqueous solution 100% Zymomonas liquid culture extract b ) 50% aqueous solution 100% Zymomonas liquid culture extract b) 25 aqueous solution 100% Zymomonas liquid culture extract c) 100% aqueous solution 95% Zymomonas liquid culture extract c) 50% aqueous solution 95% Zymomonas 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 liquid culture extracts a) to c) was evaluated by measuring the hydration state of the stratum corneum in the living body [SKI].
CON 200 (manufactured by IBS Inc., Hamamatsu City)]
Was measured using. A sodium hyaluronate aqueous solution, which is recognized in the art as having moisture retention, was used as a comparative example, and distilled water was used as a control example. The previous period equipment is 3.5MHz
The moisturizing effect of the stratum corneum of the skin epidermis is measured by measuring the electric resistance of the stratum corneum of the skin epidermis.

The test was carried out by a total of 10 female and 5 males. Any 6 points (about 2 x 2 cm) on the inner side of each left and right forearm of each subject were divided (12 for each subject).
Location), and Zymomonas bacterium liquid culture extract a) to
c), an aqueous solution of sodium hyaluronate (0.075 wt%), distilled water was applied, and the remaining one area was an area for blank test application.

The moisturizing condition of each area before coating was measured by the above hydration condition measuring device, and these measured values were designated as M ₀ .
After that, 500 μl of each sample was applied to each area, and after application 3
The moisturized state after 0 minutes was measured, and these measured values were defined as M ₃₀ . Table 2 shows the moisturizing effect value obtained by the following formula from the obtained average values of M ₀ and M ₃₀ .

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

[Table 2] Table 2 Moisturizing effect of liquid culture extract of Zymomonas ──────────────────────────────────── Sample Moisturizing effect (%) ─────────────────────────────────── Zymomonas liquid culture extract a) 72 .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,
Each of the 30 females (aged 30 to 45) was subjected to a usage test for 1 month as a control. After use, the moisturizing effect was judged by a questionnaire survey on moist feeling, smooth feeling, refreshing feeling and sticky feeling. As a result, the cosmetics of the present invention showed remarkably excellent moisturizing effect as compared with the comparative cosmetics (Table 3, Table 4, Table 5 and Table 6).

[0041]

[Table 3] Table 3 Moisturizing effect of lotion of the present invention (number of questionnaire results) ─────────────────────────────── ────── Lotion of the present invention (Example-1) Evaluation ────────────────────────── Very good Good Normal ─ ─────────────────────────────────── Moisture 20 7 3 Smooth 20 8 2 Refresh 19 6 5 Not sticky 20 6 4 ──────────────────────────────────── (the following margin)

[0042]

[Table 4] Table 4 Moisturizing effect of conventional lotion (number of questionnaire results) ──────────────────────────────── ───── Conventional lotion (Comparative example-1) Evaluation ────────────────────────── Very good Good Normal ─── ───────────────────────────────── Moisture 1 8 21 Smooth 2 9 19 Refresh 2 9 19 Not sticky 1 7 22 ────────────────────────────────────

[0043]

[Table 5] Table 5 Moisturizing effect of the emulsion of the present invention (number of questionnaire results) ──────────────────────────────── ───── Emulsion of the present invention (Example-3) Evaluation ────────────────────────── Very good Good Normal ─── ───────────────────────────────── Moisture 19 8 3 Smooth 16 11 3 3 Refresh 22 5 3 Not sticky 20 4 4 ──────────────────────────────────── (the following margin)

[0044]

[Table 6] Table 6 Moisturizing effect of conventional emulsion (number of questionnaire results) ───────────────────────────────── ──── Conventional emulsion (Comparative example-2) Evaluation ────────────────────────── Very good Good Normal ───── ─────────────────────────────── Moisture 2 10 20 Smooth 4 8 18 Refresh 3 9 18 Not 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 at the same time, it contains a viscous substance which enhances the skin moisturizing effect. Therefore, the cosmetic containing Zymomonas cultures provides an extremely excellent cosmetic having an excellent effect of suppressing lipid peroxide production in the skin by ultraviolet rays and a moisturizing effect, and the culture is industrially and stably produced. It also has a remarkable effect of achieving the object of the invention.

Claims (2)

[Claims] 1. A cosmetic comprising a culture of Zymomonas bacterium. 2. A lipid peroxide production inhibitor, which comprises a culture of Zymomonas bacterium.