JP5715666B2 - Cosmetic composition - Google Patents

Description

  The present invention relates to a cosmetic composition characterized by enhancing the moisturizing effect of novel skin cells.

  The epithelial tissue in the skin is positioned at the forefront with the outside world, and plays a role of a barrier function that protects itself from various stresses by tightly binding cells. Filaggrin is a protein produced from epidermal cells and plays an important role in the formation of the stratum corneum that acts as a barrier function of the skin.

  Skin diseases are caused by physical stress due to UV exposure and friction, biological stress due to bacterial infection, chemical stress due to exposure to pharmaceuticals, chemical substances, and the like. Skin diseases are characterized by a decrease in filaggrin production in epidermal cells due to these stresses, a breakdown of the barrier function due to weakening of cell-cell attachment, and a continued decrease in skin condition due to a decrease in skin cell water content. It is a disease.

  Examples of skin diseases include atopic dermatitis, dry skin, sensitive skin, rough skin, and pimples. In the treatment of these skin diseases, after the emergency treatment of the affected area, the body is generally cured by the resilience and natural healing power. However, since natural healing requires a long period of time to recover, and because of the persistence of pain and the effects of bacterial infections, so far, improving the resilience by directly applying therapeutic drugs, reducing pain, bacteria Treatments such as prevention of infection or reduction of inflammation have been performed.

  As a drug for suppressing the symptoms of such skin diseases, there are methods of directly applying a topical steroid drug or a non-steroid topical drug to the affected area. Brednisolone, diflorazone acetate, betamethasone valerate, fluocinolone acetonide, dexamethasone, fluquinonide, alclomethasone propionate, and non-steroidal topical drugs bufexamac, bendazac, and suprofen are known as topical steroids. However, side effects such as fever, chronically progressing osteoporosis and arteriosclerosis have been reported when topical steroid drugs are used extensively in the body for a long time.

  Non-steroidal topical anti-inflammatory analgesics have a weaker anti-inflammatory action than steroids and are often used in mild cases with few side effects, but may cause a strong rash and irritation. Therefore, these drugs are not always safe.

  As cosmetic compositions having a moisturizing effect, glycerin, urea, amino acids or derivatives thereof, and hyaluronic acid are known. However, glycerin, urea, amino acids or derivatives thereof, and hyaluronic acid may impair the feeling of use of the product if the blending amount is increased in order to obtain a sufficiently high effect.

  Under these circumstances, attention has been focused on cosmetic compositions using lactic acid bacteria or cultures that are considered to be safer and more effective. As a cosmetic composition using lactic acid bacteria, proteins derived from lactic acid bacteria that adhere to skin cells antagonistically inhibit the adhesion of harmful bacteria (Patent Document 1), and anti-inflammatory active ingredients derived from lactic acid bacteria cause inflammation. It is known to suppress (Patent Document 2). However, it has a limited effect on inhibiting the adherence of harmful bacteria to the epidermis and does not lead to a radical cure of inflammatory symptoms, or a sufficient effect cannot be expected unless it is used in a large amount. There is.

  Further, a cosmetic composition for preventing and / or treating sensitive skin or dry skin is disclosed (Patent Document 3), and the use of probiotic microorganisms is described, and Lactobacillus crispatas is also exemplified. However, there is no specific description to the extent that sufficient effects can be exhibited.

By the way, Lactobacillus crispatus (Lactobacillus crispatus) KT-11 strain (FERM BP-11332) is a human-derived lactic acid bacteria. The present applicant has patented found that already with the antiallergic effect by ingesting Lactobacillus crispatus (Lactobacillus crispatus) KT-11 strain (FERM BP-11332) orally, safety It has been confirmed (Patent Document 4).

Japanese Patent No. 4726109 Special Table 2006-519014 Special table 2009-503042 Japanese Patent No. 4921499

  An object of the present invention is to provide a cosmetic composition and a cosmetic that are superior in moisturizing effect to skin cells than conventional lactic acid bacteria and are also excellent in safety.

The present invention, microorganisms or culture result of intensive studies to achieve the above object, quite surprising that Lactobacillus was enzyme treated crispatus (Lactobacillus crispatus) KT-11 strain (FERM BP-11332) is It has been found that it has an excellent moisturizing effect on skin cells and has been completed. That is, the present invention relates to a cosmetic composition having the following constitution.

(1) Lactobacillus crispatus (Lactobacillus crispatus) KT-11 strain cosmetic composition, characterized in that the cells or the culture solution is obtained by treatment with enzymes (FERM BP-11332).
(2) The cosmetic composition according to (1), wherein the enzyme is a protease.
(3) The cosmetic composition according to (2), wherein the protease is papain.
(4) The cosmetic composition according to (1), (2) or (3), wherein the moisturizing effect of skin cells is enhanced.
(5) A cosmetic for improving skin diseases, comprising 0.01 to 10% by dry weight of the cosmetic composition according to any one of (1) to (4).
(6) The cosmetic for improving skin diseases according to (5), wherein the skin disease is any one of atopic dermatitis, dry skin, sensitive skin, and rough skin.

The present invention is characterized in that using the cells or the culture solution of the particular KT-11 strain Among the Lactobacillus crispatus (Lactobacillus crispatus) (FERM BP- 11332), yet treated with an enzyme.

  Therefore, as shown in Example 1, the difference in effect between the non-enzymatic treatment described in Patent Document 3 and the present invention is extremely large.

  The cosmetic composition according to the present invention is effective in improving the skin condition such as atopic dermatitis, rough skin, dry skin, sensitive skin, etc., and can provide a safe cosmetic without side effects.

The figure which shows the filaggrin production ability in a human epidermis cell. The figure which shows the skin moisturizing effect of the skin lotion which mix | blended this invention product. The figure which shows the improvement effect of atopic dermatitis.

Although an example of the method of obtaining the cosmetic composition used in the present invention is shown, it is not particularly limited thereto.
Lactobacillus crispatus used in the present invention (Lactobacillus crispatus) KT-11 strain (FERM BP-11332) is a human-derived lactic acid bacteria, the strain has been deposited with the National Institute of Technology and Evaluation. The bacteriological (morphological, cultural, physiological) properties of the strain are as follows.

[1] Morphological properties
In observation after culturing at 37 ° C. for 72 hours on MRS agar medium (Difco), the size of the cell is a gonococcus with 0.5 to 1 × 3 to 5 μm and does not move. There is no sporulation and Gram staining is positive.

[2] Culture characteristics
The colonies after culturing at 37 ° C. for 72 hours on MRS agar medium (Difco) have a diameter of 2 to 3 mm, are circular, and have a full edge. The color of the colony is yellowish white and translucent.

[3] Physiological properties Gas production Negative glucose utilization Positive catalase activity Negative gelatin liquefaction Negative nitrate reducing ability Negative indole productivity Negative hydrogen sulfide productivity Negative oxygen attitude Facultative anaerobic optimal growth temperature 37-40 ℃
Optimum growth pH pH 5.5-5.8

As a medium for culturing Lactobacillus crispatus (Lactobacillus crispatus) KT-11 strain (FERM BP-11332), a medium usually used for culture of lactic acid bacteria is used. That is, any medium can be used as long as it contains a nitrogen source, an inorganic substance, and other nutrients in addition to the main carbon source. As the carbon source, glucose, lactose, sucrose, fructose, starch hydrolyzate, molasses, etc. can be used. As the nitrogen source, casein hydrolyzate, soybean protein hydrolyzate, potato hydrolyzate, organic nitrogen-containing substances such as shochu can be used. In addition, meat extract, fish extract, yeast extract, oleic acid and the like are used as growth promoters.

  The culture may be performed by a method used for normal lactic acid bacteria culture. It can be performed under either aerobic or anaerobic conditions. In general, the culture temperature is preferably 37 ° C., but other temperature conditions may be used as long as the bacteria grow. It is desirable to maintain the pH of the medium during the culture at 5.0 to 5.5, but other pH conditions may be used as long as the bacteria grow. The culture time is usually preferably 12 to 24 hours, but any other culture time may be used as long as the bacteria can grow.

  The enzyme-treated product used in the present invention can be obtained by treating bacterial cells or a culture solution with an enzyme. A culture solution is desirable for the treatment of the enzyme, but it may be a cell, a cell suspension, a culture supernatant, a freeze-dried powder of the culture, or the like. As the enzyme used in the present invention, a protease is preferable, and an endo-type protease is particularly preferable. Examples of the endo-type protease include aspartic type protease, metallotype protease, serine type protease, and cysteine type protease. Of these, cysteine type proteases are more preferred, and cysteine type proteases include papain, bromelain, ficin and the like. Of these, papain is most preferable, but any enzyme capable of completely or partially degrading bacterial cells or culture solution may be used, for example, lipase, actinase, lysozyme and the like.

Enzymatic treatment method of Lactobacillus crispatus (Lactobacillus crispatus) KT-11 strain (FERM BP-11332) varies depending on the type of enzyme, for example, when using the enzyme papain, papain enzyme against culture solution or fungus body It may be added at 0.05% and reacted at 37 ° C. for 5 hours, and the enzyme treatment conditions can be appropriately selected. The obtained enzyme-treated product is preferably heated at 100 ° C. or higher for several seconds after the reaction to inactivate the enzyme, but may not be heated. The obtained enzyme-treated product can be used as a liquid, but may be used after being dried by a freeze drying method or a spray drying method.

  In this specification, the term “cosmetics” refers to a preparation used for the skin and mucous membrane that achieves a desired effect by being brought into contact with epithelial tissues such as skin and mucous membrane. In particular, the present invention is effective for use in contact with skin or mucous membranes continuously for a long time.

  The blending amount of the cosmetic composition according to the present invention cannot be generally determined, but the preferable blending amount is 0.001 to 90% as a dry weight with respect to the cosmetic, more preferably 0.01 to 10%. It is.

  The cosmetic composition according to the present invention is blended in cosmetics, but the cosmetics to be blended are not particularly limited. For example, lotion, cosmetic liquid, emulsion, cream, foundation, eye shadow, lipstick, blusher , Emollient cream, emollient lotion, cream, cream rinse, cold cream, vanishing cream, lotion, pack, gel, face pack, soap, body soap, shampoo, conditioner, rinse, face wash, shaving cream, hair cream, hair lotion And cosmetics for skin or hair such as hair treatment, hair pack, gloss, lip balm, cake, etc., and oral cosmetics such as toothpaste, wet toothpaste, liquid toothpaste, mouthwash, and mouth freshener.

  Cosmetics are also included in the cosmetics. Cosmetics include cleansing cosmetics, hair cosmetics, basic cosmetics, makeup cosmetics, aromatic cosmetics, tanning cosmetics, sunscreen cosmetics, nail cosmetics, eyeliner cosmetics, eyeshadow cosmetics, teak, lip cosmetics, oral cosmetics, etc. The present invention is effective for any application.

  Furthermore, the cosmetic may be a pharmaceutical or a quasi drug. For example, the cosmetic composition according to the present invention can be blended in an ointment containing a pharmaceutically active ingredient. Preferred examples of the external preparation for skin include skin lotion, cosmetic liquid, emulsion, cream, foundation, eye shadow, lipstick, blusher, emollient cream, emollient lotion, cream, cream rinse, cold cream, vanishing cream, lotion, pack, Gel, face pack, soap, body soap, shampoo, conditioner, rinse, face wash, shaving cream, hair cream, hair lotion, hair treatment, hair pack, gloss, lip balm, cake, sunscreen cosmetics, hair cosmetics and oral cavity Cosmetics are listed.

  Examples of the dosage forms of the cosmetics of the present invention include ointments, thickening gel systems, lotions, tablets, water-in-oil emulsions, oil-in-water emulsions, solids, sheets, powders, gels, mousses and Examples include sprays. A product in a form in which a cosmetic composition according to the present invention is impregnated into a cloth, such as a make-up pack, may be used.

  Furthermore, the cosmetic of the present invention can be used not only for humans but also for pets, livestock, poultry, racehorses, other mammals and fish.

  The cosmetics according to the present invention can be used by a method similar to the commonly used method. The cosmetic according to the present invention can be used at any frequency, for example, once a day, twice a day, or three times a day. The application site of the cosmetic according to the present invention is arbitrary, and may be applied to the skin at any site in the whole body. Examples of application sites include the face, head, oral cavity, arms, hands, legs, chest, back, and the like.

  Although the composition example or Example applicable to the cosmetics of this invention is shown below, the characteristic and the outstanding effect of this invention are demonstrated concretely, This invention is not restrict | limited by this Example.

[Example 1] shown below filaggrin production ability by cultures of human epidermal cells Lactobacillus was treated with studied enzyme of filaggrin production ability in crispatus (Lactobacillus crispatus) KT-11 strain (FERM BP-11332).

Lactobacillus crispatus (Lactobacillus crispatus) KT-11 strain (FERM BP-11332) was inoculated to the medium shown in Table 1, it was cultured for 18 hours at 37 ° C.. After culturing, the cultivated product was heat-treated at 110 ° C. for 15 minutes and freeze-dried. Further, the culture was reacted with 0.05% papain at 37 ° C. for 5 hours, and then heat-treated at 110 ° C. for 15 minutes to inactivate papain was used as an enzyme-treated culture.

Normal human epidermal cells suspended in Humedia-KG2 medium were seeded in a 96-well plate at 1 × 10 ⁴ cells / mL per well, and cultured overnight at 37 ° C. in the presence of 5% CO ₂ . After culturing, replace with medium supplemented with culture or enzyme-treated culture to a concentration of 0.1%. After 48 hours of culturing, RNA is recovered using FastLane Cell cDNA Kit, converted to cDNA, and filaggrin production is determined by real-time PCR. It was measured. GAPDH was used as a control housekeeping gene.

FIG. 1 shows the relative value of filaggrin production in human epidermal cells. Cultures of the enzyme treated with Lactobacillus crispatus (Lactobacillus crispatus) KT-11 strain (FERM BP-11332) was more significantly filaggrin production ability as compared to cultures not treated with enzyme. These results, Lactobacillus crispatus (Lactobacillus crispatus) KT-11 strain (FERM BP-11332) filaggrin production ability of the cultures by the treatment with enzyme was found to significantly enhance.

Example 2 The amount of Lactobacillus crispatus the study was the enzymatic process of Example 1 (Lactobacillus crispatus) KT-11 strain (FERM BP-11332) below moisturizing effect on the skin by aqueous glycerin solution containing a combination of culture Shown in

A lotion comprising a 20% aqueous glycerin solution containing 0 to 20% concentration of the enzyme-treated culture of Example 1 was prepared. To 3 volunteers, 10 μL of lotion was applied to 2 cm ² of the skin under the elbow and held for 5 minutes. After holding, excess moisture was wiped off with a Kim towel, and the moisture content of the skin was measured using a moisture checker.

  FIG. 2 shows the moisturizing effect on the skin of the skin lotion. The moisture content of the skin to which the skin lotion containing 0.01 to 10% enzyme-treated culture was applied was significantly higher than that of the skin lotion to which no enzyme-treated culture was added. From the above results, it was found that the moisturizing effect on the skin was increased by adding 0.01 to 10% concentration of the enzyme-treated culture to the skin lotion.

Example 3 Lotion according to the enzymatic treatment of the improvement embodiment 1 of drying sensitive skin Lactobacillus crispatus (Lactobacillus crispatus) KT-11 strain (FERM BP-11332) drying by lotion containing cultures The effect of improving sensitive skin is shown below.
Table 2 shows the composition of the lotion used in the test.

A skin lotion containing the enzyme-treated culture of Example 1 was applied to 30 women with dry sensitive skin symptoms for 8 weeks to verify the improvement effect of dry sensitive skin. An appropriate amount of lotion was applied to the affected area having symptoms of dry sensitive skin twice a day in the morning and before going to bed. The effect of improving the affected area was evaluated by a questionnaire survey using the VAS method in which the most severe condition was recorded as 10 and the absence of any condition was recorded as 0. Significant differences were obtained by comparing each value with that of week 0 by Wilcoxon signed rank sum test.
Table 3 shows the questionnaire results regarding skin symptoms.

From the results in Table 3, it is understood that “itchiness”, “dryness”, “redness”, “smooth feeling”, and “moist feeling” were remarkably improved at 4th and 8th weeks compared to 0th week.
These results, Lactobacillus crispatus (Lactobacillus crispatus) in which the enzyme treatment of Example 1 KT-11 strain (FERM BP-11332) lotion containing cultures coating is effective to improve the drying properties sensitive skin It can be seen that it is.

[Example 4] Lotion by Lactobacillus crispatus that the enzymatic treatment of improvement in Example 1 of atopic dermatitis (Lactobacillus crispatus) KT-11 strain (FERM BP-11332) Atopic by lotion containing cultures The improvement effect of atopic dermatitis is shown below.

The lotion of Example 3 was used for the test. To five women with atopic dermatitis symptoms, lotion was applied for 12 weeks to verify the improvement effect of atopic dermatitis. That is, an appropriate amount of lotion was applied twice a day to the affected area having atopic dermatitis symptoms in the morning and before going to bed. The severity of atopic dermatitis was determined by visual assessment by a dermatologist as 1 point for asymptomatic, 2 points for mild, 3 points for moderate, and 4 points for altitude.
Table 4 shows the severity assessment of atopic dermatitis.

In the items of `` dry '', `` scales / desquamation '', `` itchiness '', `` crack '', `` general severity ''
It can be seen that the symptoms improved remarkably at 6 and 12 weeks compared to 0 week.

  FIG. 3 is a photograph of different cases with improved atopic dermatitis. In each case, the symptoms of atopic dermatitis in the neck, wrist, and elbow were significantly improved at 6th and 12th weeks compared to 0th week.

These results, Lactobacillus crispatus (Lactobacillus crispatus) in which the enzyme treatment of Example 1 KT-11 strain (FERM BP-11332) coated culture lotion containing the effective improvement of atopic dermatitis It can be seen that it is.

Claims (2)

Lactobacillus crispatus (Lactobacillus crispatus) KT-11 strain cosmetic composition, characterized in that the cells or the culture solution is obtained by treatment with the enzyme papain (FERM BP-11332). 2. The cosmetic composition according to claim 1, which is used for improving atopic dermatitis or dry sensitive skin.

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