CN1638784A - skin topical composition - Google Patents

Abstract

Disclosed is a composition for external application to the skin, which comprises a fermented milk product obtained from Kefia grain and a pharmaceutically or cosmetically acceptable carrier; a method of treating skin using the above composition; the application of the composition; cell activator, skin aging inhibitor, whitening agent or melanin generation inhibitor containing fermented milk product obtained from kefir milk granule is provided. The above-mentioned composition for external application to the skin contains, as an active ingredient, a fermented milk product obtained from kefir grains, preferably kefir and/or kefir whey. These skin external compositions are very suitable for use as cosmetic compositions, scalp/hair care compositions, dermatological treatment compositions, and the like, because of having various advantageous effects of activating cells, preventing skin aging, whitening, inhibiting melanogenesis, and the like.

Description

skin topical composition

technical field

The present invention relates to an external skin preparation containing a fermented milk product obtained from kefir kefir granules. In particular, the present invention relates to an external skin preparation containing kefir and/or kefir whey kefir granules, a fermented dairy product obtained therefrom, from kefir granules. The skin external preparation of the present invention is very suitable as a cosmetic product, a composition for the scalp and hair, a composition for cleansing the skin, a composition for treatment of skin diseases, etc., which has an excellent cell activation effect, Anti-aging effect, whitening effect, effect of inhibiting melanin production, etc.

Background technique

Skin activity is significantly affected not only by the internal environment such as aging, but also by the external environment such as ultraviolet rays and various stimuli, thereby easily causing some skin problems such as rough skin, wrinkles, spots, and dull skin. In order to alleviate these problems, a large number of skin-activating ingredients have been tested. Among them, a lot of research has been done on cell activation components and whitening components. Cell activating ingredients can stimulate cell division, enhance metabolism and have a good effect on maintaining body health. Also, the ingredients activate epidermal keratinocytes, exfoliate aged keratinocytes and promote a keratinization process called skin renewal, thereby improving the skin to a healthy, aesthetically pleasing state.

Kefir is a traditional fermented dairy product from the Caucasus region. It is a drink similar to yogurt obtained by fermenting mammalian milk using a seed strain called kefir granule as the mother bacteria. In particular, the clear liquid part of this yogurt-like fermented dairy product is called kefir. Kefir can be in a dry state from which almost all the water has been removed, or in a gel state with relatively more water. A method for producing dry kefir milk is disclosed in, for example, JP-83842/1987 publication and a method for producing kefir whey is disclosed in JP-219538/1990 publication. The disclosure content of the above-mentioned documents is hereby incorporated by reference.

In addition, many people are keen on the research of kefir, not only to obtain healthy food, but also because of the application of drugs focused on its excellent health effects (JP-320069/1993 communique, JP-211668/1994 communiqué, JP-37711/1997 communique , JP-191852/1997 bulletin, etc.). Most of the research and development carried out is aimed at the oral ingestion of food or pharmaceutical compositions containing kefir milk.

Although kefir has excellent effects, it is difficult to use in addition to oral products compared to other common yogurt products due to its original taste and pH characteristics (about 4.20-4.40) as a fermented milk drink. Other aspects, such as skin topical preparations, especially cosmetic aspects that require a better touch when applied.

On the other hand, the concept of health is being multifaceted and beginning to include not only mental and physical aspects but also appearance, such as skin, belonging to the field of beauty. In order to maintain health in an extended sense, in addition to drugs expected to have a quick effect, various slow-acting but safe and easy-to-use products are also required in daily life.

Contents of the invention

In view of these circumstances, the present inventors have been focusing on the functions of fermented milk products obtained from kefir granules as a traditional health-enhancing food, namely, safety, thermal stability, and effects on skin such as cell activation, and by overcoming the above-mentioned defects and utilizing The properties of said fermented milk products complete the present invention.

In other words, the present invention relates to a composition of an external preparation for skin comprising a fermented milk product obtained from kefir granules, and a pharmaceutically and cosmetically acceptable carrier thereof. Likewise, the present invention relates to a composition for external use on skin, such as a cosmetic composition, a composition for scalp and hair, and a composition for skin cleanser, further comprising one or more selected from the following Ingredients: free radical scavenging ingredients, antioxidant ingredients, anti-inflammatory ingredients, UV protection ingredients and moisturizing ingredients. Furthermore, the present invention relates to a cell activator, an antiaging agent, a skin whitening agent or a melanin production inhibitor containing the fermented milk product of the present invention.

Also, the present invention relates to methods for activating skin cells, anti-aging skin, inhibiting skin melanin biosynthesis, and/or whitening skin by applying to skin an effective amount of fermented milk product obtained from kefir granules. Preferred embodiments of the method of the present invention include a method for making up or cleansing the skin, a method for making up or cleansing the scalp and/or hair, a method for treating the skin of a dermatological patient, and the like.

Furthermore, the present invention relates to the use of a fermented milk product obtained from kefir granules in the production of a skin external preparation composition.

Description of drawings

FIG. 1 is a photograph as a graph showing the results of Experimental Example 1. FIG.

Fig. 2. is a graph showing the results of Experimental Example 1 (hMLH1).

Fig. 3. is a graph showing the results of Experimental Example 1 (hPMS1).

Fig. 4. is a graph showing the results of Experimental Example 1 (hPMS2).

Fig. 5. is a photograph as a graph showing the results of Experimental Example 2.

Fig. 6. is a graph showing the results of Experimental Example 2 (hMLH1).

Fig. 7. is a photograph as a graph showing the results of Experimental Example 3.

Fig. 8. is a graph showing the results of Experimental Example 3.

Fig. 9. is a photograph as a graph showing the results of Experimental Example 4(1).

Fig. 10. is a graph showing the results of Experimental Example 4(1).

Fig. 11. is a photograph as a graph showing the results of Experimental Example 4(2).

Fig. 12. is a graph showing the results of Experimental Example 4(2).

Fig. 13. is a photograph as a graph showing the results of Experimental Example 4(3).

Fig. 14. is a graph showing the results of Experimental Example 4(3).

Fig. 15. is a graph showing the results of Experimental Example 5.

Best Mode for Carrying Out the Invention

The fermented milk product obtained from kefir granules used in the present invention can be obtained by fermenting mammalian milk, soybean milk, powdered skim milk, etc., with granular bacteria called kefir The seed strain of milk granules, which contains at least 5-6 combined functions such as lactic acid, yeast, etc. When kefir particles are inoculated into mammalian milk such as cow's or goat's milk, soybean milk, skim milk, etc., lactic acid fermentation by lactobacillus and ethanol fermentation by yeast can occur. The fermented milk product of kefir granules is different from yogurt fermented only by lactobacillus, has a sour taste, and has a pungent smell in addition to the slight alcohol taste caused by the carbon dioxide gas released by the yeast. Also, said fermented milk products are rich in nutrients such as lactose, peptides, amino acids, lactic acid, vitamins, minerals to bring good effects to our body.

The fermented milk product used in the present invention can be obtained by fermenting common mammalian milk. Examples of mammalian milk include cow's milk, mare's milk, sheep's milk, goat's milk, buffalo's milk, and the like. The method of producing fermented products from mammalian milk is not limited to but can be a traditional method, such as using ordinary cow's milk as raw material and fermenting with kefir granules, or using mare's milk which is more common in Eastern Europe. However, it is preferred to use cow's milk as a raw material and obtain the product fermented with kefir granules (see JP-83842/1987 communique).

In addition, ordinary soybean milk, powdered skim milk, etc. can be used to produce fermented milk products through kefir granules.

For the kefir and kefir whey in the present invention, the above-mentioned fermented milk products similar to yoghurt or their dry components are called kefir, and from the yoghurt-like The resulting transparent liquid component is called kefir whey. A preferred embodiment of the fermented milk product obtained from kefir granules used in the present invention may be kefir or kefir whey, or a mixture of both.

The fermented milk product obtained from kefir granules used in the present invention, kefir exhibits various uses, including moisturizing effect, ultraviolet protection effect, melanin production inhibiting effect, free radical removal effect and cell activation effect, etc. . Since it also exhibits heat stability and is filter sterilizable, it can be used in stable external preparations for the skin, especially in cosmetic materials. Although it has a unique smell produced by yeast fermentation and lactic acid fermentation and a pH characteristic of 4.20-4.40, by mixing with other cosmetic raw materials or carriers, it is expected to have a better feeling in use and show beneficial effects and utility.

The kefir used in the present invention can be in a form similar to yogurt, or in the form of a powder obtained by drying (freeze-drying, etc.) kefir similar to yogurt, or by further processing the powdered kefir And the obtained granular. It may also be in a liquid state obtained by treatment with water, buffer, alcohol, oil, etc.

When the kefir is used in an external preparation for skin, it preferably contains 0.01-3.00 wt% kefir, preferably 1.00-3.00 wt%, based on the total weight of the external preparation.

In addition, the kefir whey used in the present invention is a transparent liquid component obtained from the above-mentioned fermented milk products similar to yoghurt, also known as "milk serum". It is rich in nutrients such as lactose, peptides, amino acids, lactic acid, vitamins, minerals; and is considered to contain about half of the nutrients in raw milk. In kefir whey, the solids content is about 6%, and the rest is water content. Kefi whey contains more L-lactose than ordinary yogurt, and the antibacterial effect of Lactobacillus is attracting attention. Also, it has been found that kefir whey maintains skin moisture and adjusts pH to ideal mild acidity because kefir whey has a similar structure to NMF (natural moisturizing factor) in skin keratin cells. In addition, kefir milk is considered to protect the skin from external stimuli or bacteria due to its anti-allergic effect and excellent moisturizing effect.

Despite these beneficial properties, kefir whey has been viewed as little more than a waste product. Only recently has its effective use been discussed.

The dairy product kefir whey obtained by fermenting kefir granules used in the present invention exhibits various useful effects such as the same moisturizing effect as the above-mentioned kefir, UV protection effect, melanin production inhibiting effect, free radical scavenging effect and cell activation. It also exhibits heat stability and is filter sterilizable, allowing it to be used as a stable cosmetic raw material. It has a unique smell produced by yeast fermentation and lactic acid fermentation, and a unique pH of 4.20-4.40, however, kefir milk is mixed with other cosmetic raw materials or carriers, it is expected to achieve a better feeling in application, and Can exhibit beneficial effects and utility.

When the kefir whey is used in an external preparation for skin, its preferred content is 0.01-3.00 wt% based on the total weight of the external preparation, especially, the preferred content is 0.01-3.00 wt%.

The above-mentioned fermented milk product obtained from kefir granules, whether it is kefir or kefir whey, can be used as a cell activator because it has the cell activation effect as described above. In addition, it is also useful as an anti-aging skin agent and whitening agent due to its UV protection effect, melanin production inhibiting effect, and free radical scavenging effect, and it is apparently useful as a melanin production inhibitor.

The skin external preparation composition of the present invention, when used as a drug, is a composition comprising a fermented milk product obtained from kefir granules and a pharmaceutically acceptable carrier. When used as a cosmetic product or cleanser, it is a composition comprising a fermented milk product obtained from kefir granules and a cosmetic carrier. The external preparation composition for skin of the present invention may contain any other desired ingredients used in external preparations for skin in general cosmetic products, quasi-drugs or medicines, for example, one or more ingredients selected from the following: the content range does not interfere with the present invention Effective radical scavenging ingredients, antioxidant ingredients, anti-inflammatory ingredients, UV protection ingredients and moisturizing ingredients.

The skin external preparation composition of the present invention can be properly mixed with various required carriers, oily substances, wetting agents, surfactants, antibacterial agents, fragrances, water, alcohols, other alkalis, thickeners, etc. For production, the carrier includes, for example, hydrocarbons such as petrolatum or microcrystalline wax, such as jojoba oil or spermaceti, triglycerides such as tallow or olive oil, fatty acids such as stearic acid or oleic acid, such as glycerol or polyols of 1,3-butanediol.

Examples of the free radical scavenging component contained in the skin external preparation composition of the present invention include one or more of the following components: superoxide dismutase (SOD), mannitol, carotenoids, astaxanthin, rutin, and Derivatives, bilirubin, cholesterol, tryptophan, histidine, quercetin, catechin and its derivatives, gallic acid and its derivatives, glutathione and its derivatives, Scutellaria baicalensis extract, Ginkgo biloba Extract, Spatholobus Extract, Wild Hawthorn Extract, Rose Extract, Saxifrage Extract, Lemon Balm Extract, Geranium thunbergii Extract, Peony Extract, Petunia Extract, Potentilla tormentilla Extract, Siraitiagrosvenori extract, Yashajitsu extract, Lycium chinese root extract, Stevia rebaudiana extract and Rosa multiflora thunberg extract.

Examples of the antioxidant component contained in the skin external preparation composition of the present invention include one or more of the following components: mannitol, rutin and its derivatives, quercetin, phospholipids, pyridoxine, flavonoids, polyphenolic compounds, Ginkgo biloba extract, saxifrage extract, vitamin A family and its derivatives, vitamin B family and its derivatives, vitamin C family and its derivatives, vitamin D family and its derivatives, vitamin E family and its derivatives, Butylated hydroxytoluene and butyl hydroxyanisole.

Examples of the anti-inflammatory ingredients contained in the skin external preparation composition of the present invention include one or more of the following ingredients: glycyrrhizic acid and its derivatives, glycyrrhetinic acid and its derivatives, mefenamic acid and its derivatives substances, phenylbutazone and its derivatives, indomethacin and its derivatives, ibuprofen and its derivatives, ketoprofen and its derivatives, allantoin and its derivatives, guaiac Azulene and its derivatives, ε-aminocaproic acid, diclofenac sodium, tranexamic acid and its derivatives, Ashitaba (Angelicakeiskei) extract, arnica extract, aloe vera extract, Polygonum extract, turmeric Extract, Forsythia officinale extract, Phellodendron phellodendri extract, German chamomile (Matricaria chamomilla) extract, Licorice root extract, Honeysuckle extract, Water mustard extract, Symphytumofficinale extract, Acanthopanax extract, Sage tail Grass Extract, Comfrey Root Extract, Perilla Extract, White Birch Extract, Tea Extract, Calendula Flower Extract, Elderflower Extract, Typha latifolia Extract, Sapindus mukurossi Extract Extract, Artemisia Extract, Eucalyptus Extract, Japanese Angelica Extract and Milk Vetch Extract.

Examples of the ultraviolet protection component contained in the skin external preparation composition of the present invention include one or more of the following ingredients: p-aminobenzoic acid (PABA), ethyl PABA, glyceryl PABA, N,N-dimethyl PABA -Amyl, N,N-Dimethyl PABA-2-Ethylhexyl, 2-Ethylhexyl Salicylate, Ethyl Glycol Salicylate, Homomenthyl Salicylate, 2-Ethylhexyl -4-methoxy-cinnamate, ethoxyethyl-4-methoxy-cinnamate, potassium 4-methoxy-cinnamate, methyl-4-5-diisopropylcinnamate Esters, Glyceryl mono-2-ethyl-hexanoate, Di-p-methoxycinnamate, 2-Hydroxy-4-methoxy-benzophenone, 2-Hydroxy-4-methoxy - Benzophenone sulfonate, sodium 2-hydroxy-4-methoxy-benzophenone sulfonate, 2,2'-dihydroxy-4,4'-dimethoxy-benzophenone, 2,2'-Dihydroxy-4,4'-dimethoxy-benzophenone-5-sodium sulfonate, 2-4-dihydroxybenzophenone, 2,2',4,4'- Tetrahydroxy-benzophenone, 2-(2-hydroxy-5-methylphenyl)-benzotriazole, imidazole acrylate, ethylimidazole acrylate, 4-tert-butyl-4,-methoxy -Dibenzoylmethane, titanium dioxide, zinc oxide, iron oxide, cerium oxide and zirconium oxide.

Examples of the moisturizing ingredient contained in the skin external preparation composition of the present invention include one or more ingredients selected from the group consisting of mucopolysaccharides and derivatives thereof from hyaluronic acid, chondroitin sulfate, dermatan sulfate, acetyl sulfate Heparin, heparan sulfate and keratin; proteins selected from collagen, elastin, fibronectin and keratin and their derivatives and hydrolysates; selected from glycine, alanine, valine, isoleucine, serine, Threonine, Aspartic Acid, Glutamic Acid, Asparagine, Glutamine, Lysine, Hydroxylysine, Arginine, Cysteine, Cystine, Methionine, Phenylalanine Amino acid and its derivatives of amino acid, tyrosine, proline, hydroxyproline, theanine, orthinine, citrulline, and pyrrolidone carboxylate; selected from sorbitol, erythritol, maltose , maltitol, xylitol, xylose, trehalose, inositol, glucose, pentaerythritol, fructose, sucrose and its esters, and dextrin and its derivatives; mucin, D-panthenol and its derivatives , urea, phospholipids, glycolipids and N-fatty ceramide, deep sea water, honey, brown sugar extract, aloe extract, seaweed extract, quince (Cydonia oblonga) extract, North American witch hazel extract, loofah Extract, Mallow Extract, Apple Extract, Grapefruit (Citrus paradisi) Extract, European Plum (Prunus domestica) Extract, Citrus aurantifolia Extract, Citrus junos Extract, Nanjing Tilia (Tilia miqueliana) Extract Extract, Rubus extract, Sophoraflavescens extract, Zizyphus jujuba extract, Malt extract, Artemisia capillaris, Lilium candidum extract, Hops extract, Peppermint ( Mentha piperita) extract, Houttuynia cordata extract, Peony extract, Coix coix extract, Lavender extract, Oat extract, Ruscus aculeatus extract, Marshmallow extract, Poria cocos (Wolfiporia cocos) Extract, nettle extract, fennel extract, kiwi (Actinidia chinensis) extract, cucumber extract, grape extract, cactus extract, rehmannia root extract, quince extract, Cnidium officinale extract, mulberry extract, Thyme Extract, Horse Chestnut Extract, Peach Blossom (Prunus persica) Extract, Rosa damascena Extract, Apricot Tree Extract, Corn Extract, Ginger Extract, Lime Extract, Lime Extract, Strawberry Extract, Yellow Gentian Extract, Mallow Extract, Viola Extract, Burdock Extract, Lupine Extract, Carob Extract, Ivy Extract, Pine Extract, Rodgersia podophylla Extract, Burnet Extract, Miscanthus Pedicel Flower Extract, Gentiana Gentiana Extract, Hydrangea Extract, Fukutanpopodandelion Extract, Cydonia Seed Extract, Glycerin and 1,3-Butanediol.

The skin external preparation composition of the present invention may further be appropriately mixed with other ingredients having effects such as whitening effect and cell activation effect.

Examples of the above-mentioned ingredients having a whitening effect include: vitamin C and its derivatives and salts, placenta extract, licorice extract, coix extract, asparagus extract, rose fruit extract, rabus extract, Sophora radix extract Extract, Millettia reticulata (Millettia reticulata) extract, Wujia extract, rice bran extract, malt extract, viola extract, wild hawthorn extract, (white lily) Lilium candidum extract, peony extract, soybean extract , tea leaf extracts such as green tea extract, tea leaf extract and oolong tea extract, molasses extract, white pomegranate extract, beech bud extract, grape extract, hop extract, rose extract, jujube extract, Siraitia grosvenori extract Extract, Scutellaria baicalensis extract, seaweed extract and so on.

Further, examples of the above-mentioned components having cell activation effects include: vitamin A and its derivatives, citric acid, lactic acid, tartaric acid, malic acid, glycolic acid, succinic acid, serine, glutamic acid, hydroxyproline, tea Amino acid, pyrrolidone carboxylate, yeast extract, lactobacillus extract, bifidobacterium extract, fermentation metabolite extract, etc. Among these cell activating agents, vitamin A and its derivatives, citric acid, lactic acid, malic acid, serine and pyrrolidone carboxylate are especially preferred.

In addition, the skin external preparation composition of the present invention can be widely applied to cosmetic products such as cosmetics for the outer surface of the skin, compositions for scalp and hair, compositions for skin cleansers, drugs, quasi-drugs. Also, its dosage form may be any form used in conventional skin external preparations as long as it is applicable to the skin, for example, solution system, solubilization system such as skin lotion, emulsification system such as lotion and cream, powder dispersion system, Water-oil two-phase system, water-oil-powder three-phase system, dispersion liquid, ointment, gel, aerosol, etc. When preparing these compositions for external use on skin, the types and amounts of the ingredients are adjustable, and the adjustment range can be suitably determined by those skilled in the art according to conventional methods.

In addition, the skin external preparation composition of the present invention can also be optionally used as a foundation, face wipe, lipstick, lip balm, in addition to being used as a facial cosmetic product such as skin lotion, lotion, cream, and mask or foundation. , other make-up cosmetic products, cosmetic products for special purposes, cosmetic products for the body, cosmetic products for sun protection or sun protection, cosmetic products for the scalp and hair, cosmetic products for fragrance, facial cleansing cosmetic products for hair (for scalp and for hair), body cleansers, hair cleansers (for scalp and for hair), cosmetic products for shaving, cosmetic products for bathing, etc. wait.

Also, the form and use of the external preparation for skin of the present invention are not limited to the above examples.

Example

The invention will be illustrated in more detail by the following non-limiting test examples and examples.

The purpose of the following tests is to examine the various effects on cells of the ingredients contained in fermented milk products obtained from kefir granules. First, prepare samples for each assay. The kefir solids of the fermented dairy product are centrifuged and the precipitate is removed. Then, filter it with a 0.45μm filter membrane and inject it into the NAP-25 microcolumn. The part with a molecular weight of 5000 or above is the flow-through component, while the part with a molecular weight of 5000 or below is divided into 1, 2, and 3 and 4 components.

Experimental example 1: detection of cell activation by kefir milk (1)

DNA often causes mismatch mutations due to DNA replication errors or recombination, and mismatch repair genes (hMLH1, hPMS1, hPMS2, hMSH2, hMSH3, and hMSH6) that repair such mismatch mutations are known. When these mismatch repair genes are mutated, the genome becomes unstable and the cells are at risk of malignant transformation.

The fraction with a molecular weight of 5000 or above and fraction 3 samples were added to the human malignant melanoma cell line HIV-I, and cultured for 15 hours. The expression levels of mRNAs of mismatch repair genes hMLH1, hPMS1 and hPMS2 were analyzed by RT-PCR. The results confirmed an increase in expression in the cells to which the samples were added. The results are shown in Figures 1 to 4.

Experimental example 2: detection of cell activation by kefir milk (2)

Component 3 prepared in Experimental Example 1 was added to the human malignant melanoma cell line HIV-I, cultured for 5 hours, and the protein expression level of hMLH1, one of the mismatch repair genes, was analyzed by Western blotting. As a result, an increase in the expression level in cells to which kefir was added was confirmed. The results are shown in Figure 5 and Figure 6.

Experimental example 3: Detection of PCNA growth by kefir milk

It is already known that PCNA (Proliferating Cell Nuclear Antigen) is a molecule closely related to cell proliferation or repair, and in cells lacking PCNA, its repair function will be severely lacking.

Fraction 3 was added to human malignant melanoma cell line HIV-I and cultured for 15 hours. The expression level of PCNA protein was analyzed by Western blotting. The results confirmed that the expression level in the cells added with kefir milk increased. The results are shown in Figure 7 and Figure 8.

Experimental Example 4: Detection of enhancement of expression of mismatch repair gene hMLH1

(1) Inject component 3 (3ml) into a 3ml anion exchange resin DEAE column. The adsorbed substances were eluted with NaCl, desalted, and then added to the human malignant melanoma cell line HIV-I and incubated for 15 hours. It was confirmed by RT-PCR that the expression of mRNAs of the mismatch repair gene hMLH1 had an enhanced effect. The elution procedure is as follows.

Circulation components

50mM Sodium Phosphate Buffer 6ml

Rinse 50mM Sodium Phosphate Buffer 6ml

Component 1 50mM Sodium Phosphate Buffer 6ml

Component 2 50mM Sodium Phosphate Buffer 0.15M NaCl 6ml

Component 3 50mM Sodium Phosphate Buffer 0.15M NaCl 6ml

Component 4 50mM Sodium Phosphate Buffer 1M NaCl 6ml

Component 5 50mM Sodium Phosphate Buffer 1M NaCl 6ml

If the control result is assigned a value of 1, then the relative value of each sample is as follows, and the resulting figure is shown in Figure 10.

Table 1 control Circulation components rinse component 1 component 2 Component 3 Component 4 Component 5 1.00 0.97 1.06 0.79 1.02 1.64 1.94 2.5

(2) Component 3 (3ml) was injected into a 3ml cation exchange resin DEAE column. The adsorbed substances were eluted with NaCl, desalted, and then added to the human malignant melanoma cell line HIV-I and incubated for 15 hours. The mRNA expression of the mismatch repair gene hMLH1 was confirmed to have an enhanced effect by RT-PCR technique. The elution procedure is as follows.

Circulation components

50mM Sodium Phosphate Buffer 6ml

Rinse 50mM Sodium Phosphate Buffer 6ml

Component 1 50mM Sodium Phosphate Buffer 6ml

Component 2 50mM Sodium Phosphate Buffer 0.15M NaCl 6ml

Component 3 50mM Sodium Phosphate Buffer 0.15M NaCl 6ml

Component 4 50mM Sodium Phosphate Buffer 1M NaCl 6ml

Component 5 50mM Sodium Phosphate Buffer 1M NaCl 6ml

The control result is assigned a value of 1, and the relative values of each sample are as follows, and the resulting figure is shown in Figure 12.

Table 2 control Circulation components rinse component 1 component 2 Component 3 Component 4 Component 5 1.00 1.46 1.61 1.29 1.38 1.98 1.93 1.83

(3) Change component 3 into a hydrophobic column. The adsorbed substances were eluted with NaCl, desalted, and then added to the human malignant melanoma cell line HIV-I and incubated for 15 hours. The mRNA expression of the mismatch repair gene hMLH1 was confirmed to have an enhanced effect by RT-PCR technique. The elution procedure is as follows.

Circulation components

      100mM Sodium Phosphate Buffer

1.8M ammonium sulfate 6ml

Rinse with 100mM sodium phosphate buffer

1.8M ammonium sulfate 6ml

Component 1 100mM Sodium Phosphate Buffer

1.8M ammonium sulfate 6ml

Component 2 100mM Sodium Phosphate Buffer

1.44M ammonium sulfate 6ml

Component 3 100mM Sodium Phosphate Buffer

1.44M ammonium sulfate 6ml

Component 4 100mM Sodium Phosphate Buffer 6ml

Component 5 100mM Sodium Phosphate Buffer 6ml

If the control result is assigned a value of 1, then the relative value of each sample is as follows, and the resulting figure is shown in Figure 14.

table 3 control Circulation components rinse component 1 component 2 Component 3 Component 4 Component 5 1.00 1.05 1.15 1.05 1.73 1.41 1.31 1.17

Experimental example 5: Detection of melanin production inhibitory effect of kefir whey

The antioxidant effect of kefir whey derived from kefir milk fermentation liquid on melanin production of human melanoma cell line SK-MEL was studied. Considering that the kefir whey will be used for culturing the cell system, the samples used are prepared under the condition of osmotic pressure of 0.300 osmol/kg after neutralization.

<Experiment method>

Seed SK-MEL cells in a 5 ml dish at a cell density of 4 × ¹⁰⁵ cells/ml. After the pre-cultivation, the medium was replaced with a medium containing 0% (control), 0.1% and 1% kefir whey relative to the medium components, and the culture was continued for 24 hours. Cells were recovered by trypsinization, diluted to a total cell number of 3 x ¹⁰⁶ cells in all conditions, and recovered into 1.5 ml tubes. The tube was centrifuged (5000 rpm, 10 min), 400 μl of 1 M NaOH was added to the cell pellet, and boiled at 100° C. for 10 min. The lysates were mixed well and three aliquots (100 [mu]l) were plated in 96-well plates for each condition. Absorbance at 405 nm was then detected by a plate reader and the amount of melanin was determined.

<result>

The results are shown in Table 4 and Figure 15. It can be seen from the results that when the melanoma was cultured in the medium added with kefir whey, its melanin production was obviously inhibited.

Table 4 sample Average absorbance relative value(%) control 0.255 100 0.1% 0.219 85.9 1.0% 0.214 83.9

Experimental Example 6: Detection of the effect of improving scalp and hair

Twenty healthy subjects (12 males, 8 females) used hair care products containing kefir, a fermented dairy product, for one month. Tested by applying the product on the scalp and hair after shampooing daily, mainly in itchy areas, observing hair shine, dandruff, hair loss and scalp itching.

The result is as follows.

Hair Shine:

Improvement: 12 cases, no change: 6 cases, deterioration: 0 cases, unknown: 2 cases

Dandruff:

Significant reduction: 8 cases, no change: 10 cases, deterioration: 0 cases Unknown: 2 cases

Hair loss:

Reduction: 5 cases, no change: 12 cases, deterioration: 1 case, unknown: 2 cases

Itchy scalp:

Reduction: 9 cases, no change: 8 cases, deterioration: 1 case, unknown: 2 cases

The results show that the hair care product containing kefir milk is effective as a composition for scalp and hair, can improve hair luster, reduce dandruff, reduce hair loss, and prevent scalp itching.

Experimental Example 7: A gel containing fermented dairy product kefir (trade name: kefran) was used, and the results are as follows.

(1) 26 year old female: Of almost all the skin lotions etc. I've ever used, every time I pour them into my palms, they throb so painfully when they touch the wounds on my fingers due to atopy , until now. So when my sister recommended kefran, I was worried it would hurt me in the same way, when in fact it didn't. Of course, I can apply it on my face without any pain, so from now on I can use it without any worries.

(2) 34-year-old female: After my second child was born, I developed atopy and my skin became very sensitive. Partly due to my age, I used to be dissatisfied with skincare products that have a gentle touch on application, have been looking for a good product, and since buying kefran I have been using it now. Really very good.

(3) 23-year-old female: I used to struggle to find a basic skin care product suitable for my atopic skin, but since I used kefran, my skin has improved significantly.

(4) 45-year-old female: I am sensitive to sunlight. My face, etc. can be protected from sun damage with makeup, hats, etc. However, if the backs of my hands and arms are exposed to the sun from time to time, at night the skin turns into a goosebump and the intense itching is excruciating. When I noticed this problem, if I used kefran early, my skin recovered quickly and the severe itching was relieved.

Example 1: cream

<Recipe (weight%)>

(1) Polyoxyethylene (40E.O.)-stearic acid 2.00

(2) Glyceryl monostearate (self-emulsifying type) 5.00

(3) Stearic acid 5.00

(4) Docosanol 0.50

(5) Squalene 15.00

(6) Cetyl isooctanoate 5.00

(7) Appropriate amount of antibacterial agent

(8) 1,3-Butanediol 5.00

(9) Kefi milk filtrate 1.00

(10)Natto Extract 1.00

(11) Soybean extract 0.20

(12) Saxifrage Extract 0.10

(13) Cucumber Extract 0.10

(14) Licorice Extract 0.10

(15)Dipotassium glycyrrhizinate 0.05

(16) Mulberry extract 0.05

(17)Scutellaria baicalensis extract 0.05

(18) Keratin Hydrolyzate 1.00

(19) Appropriate amount of spices

(20) Balance of purified water

<Preparation process>

The above components (1) to (7) were dissolved under heating at 70°C. Then, (8) and a portion of (20) are heated to 70°C and added to the previously dissolved ingredients, then, while cooling, ingredients (9) to (18), (19) and the remaining portion of (20) are added, Thereby a cream is obtained.

Example 2: Lotion

<Recipe (weight%)>

(1) Glycerin 5.00

(2) 1,3-Butanediol 6.50

(3) Polyoxyethylene (20mol) sorbitan monolaurate 1.20

(4) Ethanol 8.00

(5) Kefi milk filtrate 1.00

(6) Natto Extract 1.00

(7) Antibacterial agents, antioxidants Appropriate amount

(8) Appropriate amount of spices

(9) Purified water Balance

<Preparation process>

First, the above ingredients (3), (4), (7) and (8) were mixed and dissolved, and then (1), (2), (5), (6) and (9) were mixed and dissolved. These mixed and dissolved ingredients are further uniformly mixed, thus obtaining a lotion.

Embodiment (3): emulsion

<Recipe (weight%)>

(1) Stearic acid 1.50

(2) Cetyl alcohol 0.50

(3) Beeswax 2.00

(4) Polyoxyethylene (20mol) monooleate 1.00

(5) Glyceryl monostearate 1.00

(6) Ethanol 10.00

(7) Arbutin 20.00

(8) Sodium bisulfite 0.03

(9) Propylene Glycol 5.00

(10) Kefi milk filtrate 1.00

(11) Natto Extract 1.00

(12) Soybean extract 0.20

(13) Saxifrage Extract 0.10

(14) Cucumber Extract 0.10

(15) Licorice Extract 0.10

(16)Dipotassium glycyrrhizinate 0.05

(17) Mulberry Extract 0.05

(18)Scutellaria baicalensis extract 0.05

(19) Spices Appropriate amount

(20) Antibacterial agents, antioxidants Appropriate amount

(21) Purified water Balance

<Preparation process>

The above-mentioned ingredients (7), (9) and (10) to (18) were added to (21), dissolved under heating, and kept at 70° C. to form an aqueous phase. Ingredient (19) was added to (6) and dissolved to form the alcoholic phase. Ingredients (1) to (5), (8) and (20) were mixed, dissolved under heating, and kept at 70° C. to form an oil phase. The oil phase is added to the water phase for initial emulsification, and then uniform emulsification is carried out by a homogeneous mixer. The alcoholic phase was added with stirring, and the resulting mixture was cooled to 30°C while stirring, thus obtaining an emulsion.

Embodiment 4: Gel

<Recipe (weight%)>

(1) 1,3-Butanediol 5.00

(2) Trehalose solution 3.00

(3) Kefi milk filtrate 1.00

(4) Natto Extract 1.00

(5) Seaweed Extract 1.00

(6) Urea 0.80

(7) Carboxyvinyl 0.80

(8) Glycerin 0.50

(9)L-Ascorbic acid-magnesium phosphate 0.50

(10) Potassium hydroxide 0.50

(11) Phenoxyethanol 0.30

(12) Sodium carboxymethylcellulose 0.20

(13) Soybean extract 0.20

(14) Saxifrage Extract 0.10

(15) Cucumber Extract 0.10

(16) Licorice Extract 0.10

(17)Dipotassium glycyrrhizinate 0.05

(18) Mulberry extract 0.05

(19)Scutellaria baicalensis extract 0.05

(20) Purified water 84.75

<Preparation process>

The above-mentioned ingredients (3) to (8) and (13) to (19) are uniformly dissolved in (20) to form an aqueous phase. On the other hand, (1), (2), (9) and (11) were dissolved and added to the water phase. Then, the mixture is neutralized and thickened with (10) and (12), so that a gel is obtained.

Embodiment 5: peel-off mask

<Recipe (weight%)>

(1) 95% ethanol 10.00

(2) Polyoxyethylene (15mol) oleyl ether 2.00

(3) Antibacterial agents, antioxidants Appropriate amount

(4) Appropriate amount of spices

(5) Whey 1.00

(6) Seaweed Extract 1.00

(7) Dipotassium glycyrrhizinate 0.05

(8) Polyvinyl alcohol 12.00

(9) Glycerin 3.00

(10) Polyethylene glycol 1.00

(11) Purified water Balance

<Preparation process>

An aqueous phase was prepared at 80°C from the above ingredients (5) to (11), and cooled to 50°C. Then, ingredients (1) to (4) were prepared at room temperature to form an alcohol phase, which was added to the water phase and mixed uniformly. The mixture is then cooled, so that a mask is obtained.

Example 6: Liquid Foundation

<Recipe (weight%)>

(1) Lanolin 7.00

(2) Liquid paraffin 5.00

(3) Stearic acid 2.00

(4) Cetyl alcohol 1.00

(5) 2-Ethylhexyl p-methoxycinnamate 3.00

(6) Glycerin 5.00

(7) Triethanolamine 1.00

(8)Carboxymethylcellulose 0.70

(9) Purified water Balance

(10) Titanium dioxide 8.00

(11) Micronized titanium dioxide 2.00

(12)Zinc Oxide 5.00

(13)Mica 15.00

(14) Talc 6.00

(15)Pigment 6.00

(16) Kefi milk filtrate 0.50

(17) Mulberry Extract 0.05

(18)Scutellaria baicalensis extract 0.05

(19) Spices Appropriate amount

<Preparation process>

First, the above-mentioned ingredients (1) to (5) are mixed and dissolved. Then, ingredients (10) to (15) were added, mixed uniformly, and the temperature was maintained at 70°C. Subsequently, (6) to (9) were uniformly dissolved, kept at 70° C., and added to the previous dissolved mixture, and uniformly emulsified.

After the resulting mixture was cooled, ingredients (16) to (19) were added, whereby a liquid foundation was obtained.

Sensory evaluation of the skin external preparation of the present invention

20 women were invited to use the skin external preparation containing fermented milk products obtained from kefir milk granules. The preparation was applied to the entire face after washing the face every morning and evening for 3 months, and its efficacy was evaluated by a doctor.

I. Dry skin

1. Very effective 11 cases

2. Effective in 6 cases

3. No change 5 cases

4. Deterioration 0 cases

II. Efficacy on Facial Melasma

1. Very effective in 6 cases

2. Effective in 9 cases

3. No change 4 cases

4. 1 case of deterioration

III. Hard skin

1. Very effective 3 cases

2. Effective 8 cases

3. No change 8 cases

4. 1 case of deterioration

It is evident from the above evaluation that the skin external preparation according to the present invention is safe and effective for dry skin, blemishes and hard skin, and can increase skin cell activity.

Industrial Applicability

Fermented milk products obtained from kefir granules have a wide range of uses, including moisturizing effects, UV protection effects, melanin production inhibitory effects, and free radical scavenging effects. At the same time, it is heat stable and can be sterilized by filtration, which makes it possible to be used as a stable external preparation for skin, especially as a raw material for cosmetics. However, it has a distinctive yeasty and lactic acid fermented smell, and a pH of 4.20-4.40. It can thus be considered that, by being properly mixed with other cosmetic raw materials, it is confirmed that it has a pleasant touch when used, and has the initial effect and potency exhibited by fermented milk products obtained from kefir granules.

Claims (20)

1. a preparation for external application to skin compositions contains the fermented dairy product and the medicinal or used for cosmetic carrier that obtain from the kefir granule.

2. according to the preparation for external application to skin compositions of claim 1, further contain one or more following components: free radical scavenging component, antioxidant ingredients, antiinflammatory component, ultraviolet protection component and the component of preserving moisture.

3. according to the preparation for external application to skin compositions of claim 1 or 2, wherein the fermented dairy product that obtains from the kefir granule is that kefir and/or kefir are clear.

4. according to the preparation for external application to skin compositions of claim 3, wherein the content that kefir and/or kefir are clear is the 0.01-3.00wt% of total composition.

5. according to the preparation for external application to skin compositions of claim 1 to 4, wherein the preparation for external application to skin compositions is a liquid, emulsion, gel, emulsifiable paste, arbitrary form of paste or thin slice.

6. according to each preparation for external application to skin compositions of claim 1 to 5, wherein the preparation for external application to skin compositions is a kind of cosmetic composition.

7. according to the preparation for external application to skin compositions of claim 6, cosmetic composition wherein is a kind of cosmetic composition that is used for health.

8. according to the preparation for external application to skin compositions of claim 7, the cosmetic composition that wherein is used for health is a kind of composition of skin cleanser.

9. according to each preparation for external application to skin compositions of claim 1 to 5, preparation for external application to skin compositions wherein is a kind of compositions that is used for scalp and hair.

10. activation Skin Cell, the prevention skin aging suppresses melanin biosynthesis in the skin, and/or the method for skin whitening, and described method is the fermented dairy product that obtains from the kefir granule to the dermal administration effective dose.

11., be a kind of method that skin is made up or cleaned that is used for according to the method for claim 10.

12., be a kind of method that is used for scalp and/or hair makeup or cleaning according to the method for claim 10.

13., be a kind of method for the treatment of dermatosis patient skin according to the method for claim 10.

14. according to the method for claim 10, wherein the fermented dairy product that obtains from the kefir granule is that kefir and/or kefir are clear.

15. the application of the fermented dairy product that obtains from the kefir granule during producing the preparation for external application to skin compositions.

16. according to the application of claim 15, wherein the fermented dairy product that obtains from the kefir granule is that kefir and/or kefir are clear.

17. a cell activator contains the fermented dairy product that obtains from the kefir granule.

18. a resisting age of skin agent contains the fermented dairy product that obtains from the kefir granule.

19. a skin whitener contains the fermented dairy product that obtains from the kefir granule.

20. a Melanin inhibitor contains the fermented dairy product that obtains from the kefir granule.

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