JPH11322534A - Ceramide synthesis accelerator - Google Patents

Abstract

PROBLEM TO BE SOLVED: To produce a ceramide synthesis accelerator expected to improve rough skin and display treatment and improvement effects on various skin troubles and provided with excellent stability in the lapse of time by including a fungous culture having a ceramide synthesis accelerating function and butylene glycol. SOLUTION: This accelerator is obtained by including a fungous culture having a ceramide synthesis accelerating function [lactic acid bacteria culture (e.g. Streptococcus thermophilus or Lactobacillus bulgaricus), Bifidus bacterium culture [e.g. Bifidobacterium bifidum], mushroom cell body (e.g. Lentinus edodes or Celtis sinensis) and yeast plant (e.g. Saccharomyces cerevisiae or Endomyces magnusii) and (B) 1,3-butylene glycol. In the accelerator, the weight of the component A is 3 to 6 times that of the component B. The amount of addition is pref. 0.001 to 10 wt.% when the accelerator is added to a cultured epidermal cell system so as to accelerate a ceramide synthesis, and it is pref. 0.01 to 20 wt.% when added to agent for external use for skin.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention is expected to improve or cure rough skin and various skin diseases by activating ceramide synthesis of epidermal cells in the surface of the skin and improving the skin barrier function. The present invention relates to a ceramide synthesis accelerator having excellent stability.

[0002]

2. Description of the Related Art It has recently been known that ceramide, which is a kind of lipid, has an important physiological role although its quantity is smaller than that of glycerolipid which occupies most of the body. It is present in physiologically important sites in mammals including humans, and is known to be accumulated in the brain, liver, skin and the like.

[0003] In the skin, ceramide is accumulated particularly in the stratum corneum of the epidermis. It is synthesized and secreted by epidermal cells and is a major component of intercellular lipids that form a unique lamellar structure between cells (Lukas Landmann: Anat Embryol, 17).
8, 1-3, 1988). The stratum corneum is a series of physiological roles, including the ability to moisturize the skin and the physical protection of living organisms,
Although it has a so-called barrier function, intercellular lipids are the substance of this barrier function and play one of the most important roles in life support (Genji Imokawa: Journal of Cosmetic Society, 1
5, 4, 250-253, 1991). In this sense, it can be said that skin ceramide is one of the important substances in biological defense.

[0004] For rough and dry skin and various skin diseases,
It has been reported that the healthy formation of the stratum corneum is hindered and the barrier function is reduced. Specific examples include a decrease in the turnover of the epidermal layer due to aging of the skin surface, and rough or dry skin caused by external factors such as light, temperature, and weather conditions. This is thought to be caused by a decrease in the barrier function and a decrease in the moisturizing capacity of the skin and an increase in the amount of water transpiration (Shuichi Akasaki et al .: Journal of Nisshinkai, Vol. 98, No. 1,
41-51, 1988).

[0005] Among skin diseases, in atopic dermatitis, the barrier function is deteriorated or degraded not only in the inflamed part but also in the non-inflamed part of the patient, and ceramide in the patient's skin, in general or in a specific type, is reduced. A decrease in the content has been reported (Shin Kawashima: Journal of Koshokai, Vol. 15, No. 4, No. 261-262).
1991). In addition, changes in the amount of ceramide in the skin of patients have been reported in psoriasis (Stefania.M: Arch De
rmatol. 130, 452-456, 1994),
Again, this variation is thought to be related to barrier collapse.

[0006] With respect to skin diseases and insufficiencies caused by such deterioration or breakdown of the skin barrier function, administration of a moisturizer has been conventionally used to prevent the skin from becoming dry and moist, and to prevent eczema caused by anti-inflammatory agents. Control has been attempted. However, these methods have a temporary effect because they replenish some of the water or moisturizing components on the keratinous surface, and cannot provide sufficient moisture inside the skin continuously (Takemura Toshiyuki: Pharmacia, vol. 28, p. 1, 1992
Years), even if the temporary inflammation was suppressed, there were often problems with the persistence of the effects and side effects.

[0007] On the other hand, recently, improvement treatment of the skin has been attempted by external supplementation of ceramide, which is a main component of the barrier,
Efficacy has been reported for rough skin conditions and atopic dermatitis (Higaki Yuko et al .: Allergy Clinic, 13, 12)
No. 26-28, 1993). However, this method appears to have an early effect, but it is not sufficient in persistence of the effect, as with the moisturizers that have been used in the past, and the difference in transdermal absorption depending on the condition of the skin. However, there is a drawback that the effect cannot be sufficiently exerted due to the above-mentioned reasons.

[0008] On the other hand, a treatment for improving the skin has been attempted by increasing the ability of ceramide to be synthesized inside the tissue, rather than supplementing it from the outside, and so far cultures of yeast and the like have promoted ceramide synthesis in epidermal cells. (Japanese Patent Application Laid-Open Nos. Hei 8-217658, Hei 9-194383, and Hei 9-115236). However, these bacterial cultures are unstable, such as lees, and are susceptible to contamination by microorganisms, and it has been difficult to obtain a ceramide synthesis promoting substance that is stable over time.

[0009]

SUMMARY OF THE INVENTION In view of the above circumstances, the present inventors have proposed a method of activating ceramide synthesis of epidermal cells in the skin surface layer and improving the skin barrier function without impairing the effect of improving the skin barrier function. Intended to obtain a certain ceramide synthesis promoting substance, the solubility of the substance is high, there is an antiseptic effect,
As a result of various studies on substances whose action on cells and skin is alleviated, it was found that a bacterial culture having a ceramide synthesis promoting action and 1,3
-It has been found that a composition having a specific ratio of -butylene glycol has an effective ceramide synthesis promoting action and is excellent in stability over time, and has completed the present invention. That is, an object of the present invention is to improve the rough skin and various skin diseases by activating the ceramide synthesis of the epidermal cells within the skin surface layer and improving the skin barrier function, and it is expected to have excellent stability over time. An object of the present invention is to provide a ceramide synthesis promoter.

[0010]

SUMMARY OF THE INVENTION The above-mentioned object is achieved by the component A).
Culture with ceramide synthesis promoting action and component B)
Consisting of 1,3-butylene glycol as component A)
Is achieved with a ceramide synthesis accelerator which is 3 to 6 times the weight of component B).

[0011]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The configuration of the present invention will be described below in detail. The bacterial culture used in the present invention activates ceramide synthesis of the epidermal cells itself, and examples thereof include a lactic acid bacterial culture, a bifidobacterium culture, a mushroom cell culture, and a yeast culture.

As lactic acid bacteria, for example, Streptococcus
thermophilus, Lactobacillus bulugaricus, Lactococ
Bifidobacteria such as cus lactis include, for example, Bifido
Examples of mushroom cells such as bacterium bifidum
Lentinus edodes, Pleurotus ostrea
Examples of yeast such as tus (Hiratake) and Flammuiina velutipes (Enokitatake) include, for example, Saccharomyces cere
visiae, Endomyces magnusii and the like.

The bacterial culture is 3 to 6 times the weight of 1,3-butylene glycol used in the present invention. When the culture is less than 3 times, sedimentation occurs, and when the culture is more than 6 times, contamination by microorganisms may occur.

The ceramide synthesis promoter of the present invention may be used in the form of an external preparation for skin in addition to additives to cultured cells.
For example, ointments, creams, lotions, emulsions, packs and the like can be mentioned.

The base of the external preparation for skin may be a known base, for example, silicone oils such as methylphenylpolysiloxane, dimethylpolysiloxane and cyclomethicone, hydrocarbons such as paraffin and petrolatum, olive squalane, rice squalane , Rice germ oil, jojoba oil, castor oil, safflower oil, vegetable oils such as sunflower oil, olive oil, macadamia nut oil, waxes such as beeswax, mocrow, carnauba wax, octyldodecyl myristate, ester oils such as cetyl palmitate, cetanol, Behenyl alcohol, higher alcohols such as stearyl alcohol, cholesterol, phytosterols, sterols such as branched fatty acid cholesterol esters, processing oils such as hardened oils,
Higher fatty acids such as stearic acid, myristic acid, isostearic acid, oleic acid, iso-type long-chain fatty acids, and anteiso-type long-chain fatty acids; , Tar pigments, coloring pigments such as iron oxide, parabens, preservatives such as phenoxyethanol, anionic surfactants such as sodium cetyl sulfate, N-stearoyl-L-glutamate, glycyrrhizinate, polyoxyethylene alkyl ethers, Nonionic such as polyoxyethylene fatty acid ester, polyoxyethylene polyhydric alcohol fatty acid ester, polyoxyethylene hydrogenated castor oil, polyhydric alcohol fatty acid ester, polyglycerin fatty acid ester, modified silicone, sucrose ester, etc. Surfactants, cationic surfactants such as tetraalkylammonium salts, amphoteric surfactants such as betaine type, sulfobetaine type and sulfoamino acid type, and natural surfactants such as lecithin, lysophosphatidylcholine, ceramide and cerebroside Agents, pigments such as titanium oxide and zinc oxide, antioxidants such as dibutylhydroxytoluene, primary alcohols such as ethanol, dipropylene glycol, glycerin, propylene glycol, sorbitol, malbitol, diglycerin, sodium chloride, magnesium chloride, sodium sulfate , Potassium nitrate and other inorganic salts, sodium succinate, sodium aspartate and other organic acid salts, ethanolamine hydrochloride, ammonium nitrate, arginine hydrochloride, phosphate, citrate, acetate, carbonate, trishydroxyme Salts such as aminoaminomethane hydrochloride and diisopropylamine dichloroacetate, thickeners such as xanthan gum, carboxyvinyl polymer, carrageenan alkyl-modified carboxyvinyl polymer, chelating agents such as edetic acid, potassium hydroxide, diisopropanolamine, triethanolamine, etc. Neutralizer, biopolymers such as hyaluronic acid and collagen, chamomile, assembly, aloe, peach, carrot, horsetail, mulberry, peach leaf, sage, loquat leaf, cucumber, edible plant, hibiscus, turmeric, rosemary , Plant extracts such as licorice, serine, threonine, N-
Amino acids such as methyl-1-serine, aminobutyric acid, and hydroxyaminobutyric acid; ultraviolet absorbers such as hydroxymethoxybenzophenone sulfonate; vitamins A and B;
Vitamins such as Cs and Es can be used, but are not limited thereto.

When the ceramide synthesis promoter of the present invention is added to a cultured epidermal cell line to promote ceramide synthesis, the amount added is preferably 0.001 to 10% by weight.

The amount of the ceramide synthesis promoter of the present invention to be added to the external preparation for skin should be such that the total amount of the composition should be considered in consideration of the amount of the ceramide synthesis which is sufficiently promoted and the color and odor of the culture are hardly produced. On the basis, it is preferably 0.01 to 20% by weight, particularly preferably 0.1 to 10% by weight.

[0018]

The present invention will be described in detail below with reference to examples and comparative examples. Examples 1 to 5, Comparative Examples 1 to 5 (Lactic acid bacteria culture) 10 g of skim milk, 1 g of glucose, 0.1 g of nicotinic acid.
01g, 0.5g of yeast extract and 100m
The medium was prepared by autoclaving at 121 ° C. for 20 minutes (Skim milk was manufactured by Difco, glucose and nicotinic acid were manufactured by Kanto Chemical Co., Ltd., and yeast extract was manufactured by Asahi Breweries). Lactococcus lactis (IFO 12007) and Streptococcus cultivated in the same medium at 37 ° C for 24 hours.
thermophilus (ATCC 19254) and Lactobacillus b
ulgaricus (ATCC 11842) was inoculated at 1%. 37 ° C, 2
After static culture for 4 hours, the cells were removed by centrifugation, and the culture supernatant was treated at 80 ° C for 30 minutes to obtain a lactic acid bacteria culture. A fixed amount of each of the lactic acid bacteria culture and 1,3-butylene glycol was mixed to obtain ceramide synthesis promoters of Examples 1 to 5, respectively. The lactic acid bacteria culture was mixed with 1,3-butylene glycol, ethanol, and dipropylene glycol to obtain compositions of Comparative Examples 1 to 5. Mixing ratio is% by weight
It is.

[0019]

[Table 1]

The following tests were carried out using the ceramide synthesis accelerators of Examples 1 to 5 and the compositions of Comparative Examples 1 to 5 for stability over time, acceleration of ceramide synthesis and skin barrier recovery.

Test Example 1 Stability test with time (formulation stability) (1) Method The ceramide synthesis promoter of Examples 1 to 5 and the bacterial culture obtained in Comparative Examples 1 to 5 were placed in a sample tube.
The mixture was allowed to stand at 0 ° C., room temperature, and 0 ° C. for 3 months. (2) Results The results are shown in Table 1. As is clear from Table 1, the ceramide synthesis accelerator of the present invention (Examples 1 to 5) showed a time-stability effect.

Test Example 2 Temporal stability test (excess preservative) (1) Method In an unsterilized glass tube, the ceramide synthesis promoter of Examples 1 to 5 and the bacterial culture 20 obtained by Comparative Examples 1 to 5 were used.
Add Staphylococcus aureus (ATCC6538), Esche
richia coli (ATCC8739), Pseudomonas aeruginosa (ATCC
9027) was inoculated at 105 cells / ml, and allowed to stand at 25 ° C. for 28 days. × that did not exist
And (2) Results The results are shown in Table 1. As is evident from Table 1, the ceramide synthesis accelerator of the present invention (Examples 1 to 5) exhibited an excessive preservative power and a stability effect with time.

Test Example 3 Ceramide synthesis promotion test (1) Method (a) Cultured epidermal cells Normal human epidermal cells are commercially available (Cascad)
e Biologic). (B) Cell culture medium The medium used was MCDB153 medium to which BPE (bovine pituitary gland) was added as a growth factor. (C) Preparation of Hepes buffer 7.15 g of Hepes, 1.8 g of glucose, 0.22 g of potassium chloride, 7.7 g of sodium chloride, and 0.27 g of disodium hydrogen phosphate dodecahydrate were dissolved in purified water, and 1N After adjusting the pH to 7.4 with an aqueous sodium hydroxide solution, the volume was increased to 1 liter.

(D) Cell culture The number of normal human epidermal cells was counted in MCDB153 medium
× 10 4 cells / ml, seeded 4 ml each on a 60 mm collagen-coated plate (Falcon), 95%
In an atmosphere of air (V / V) -5% (V / V) carbon dioxide,
The cells were cultured at 37 ° C. for 5 days. Aspirate the culture supernatant,
1% by weight of the ceramide synthesis accelerator of Examples 1, 4 and 5
4 ml of the added MCDB153 medium was added to each dish. In addition, Hepes buffer solution was added as a control. 95% air (V / V) -5
Incubation was carried out at 37 ° C. for 6 days in an atmosphere of% (V / V) carbon dioxide gas. On day 6, 0.5 μCi of [14 C] -serine (American Radiolabeled Ch)
(manufactured by Chemicals Inc.) was added to the medium, and the culture was further performed for 2 days. After the culture, the cells were treated as follows.

(E) Extraction of lipid The medium supernatant was removed by suction, washed twice with 5 ml of Hepes buffer, and the cells were scraped off from the dish with a cell scraper (Sumitomo Bakelite). 1.6 ml of this
In Hepes buffer, 4 ml of methanol and 2
Add ml of chloroform and mix. After allowing to stand at room temperature for 20 minutes, a 1.6 ml chloroform layer was taken to obtain a lipid fraction. Remove chloroform by centrifugation and remove 1 ml
Redissolved in benzene.

(F) Isolation of Ceramide Fraction Using Iatrobeads Column
The solution was applied to a column packed with 1, washed with a benzene-ethyl acetate (4: 1) solution, and eluted with 1 ml of ethyl acetate to obtain a ceramide fraction.

(G) Measurement of radioactivity of [14C] -labeled ceramide The radioactivity incorporated in the ceramide fraction was measured with a liquid scintillation counter. (2) Results The results are shown in Table 2.

[0028]

[Table 2]

As is clear from Table 2, the ceramide synthesis accelerator of the present invention (Examples 1, 4 and 5) showed an effect of promoting ceramide synthesis.

Test Example 4 Skin Barrier Recovery Test (1) Method Skh: hr hairless mouse male (Japan SLC), 6 weeks old, was purchased as a test animal and preliminarily reared for 2 weeks.
The experiment was started with 5 animals per group. 20 μg of retinoic acid (vitamin A acid: all-transretinoic acid, SIGMA) for rough skin
Was dissolved in ethanol and applied to the buttocks of the mouse once a day (morning) for 3 days.

2 ml of the ceramide synthesis promoter of Examples 1, 4 and 5 was lyophilized, dissolved in the same amount of 50% (V / V) ethanol, and the same procedure was carried out on the afternoon of the day when the application of retinoic acid was started. 100 μl was applied once a day (afternoon) for 3 days. Note that the control is a sample to which only 50% (V / V) ethanol has been applied.

Three days after the application of retinoic acid, the transepidermal water loss (TEWL) was measured and expressed as the water loss (mg / cm2 / min). TEWL is an index for measuring the skin barrier function.
It rises when the barrier function is destroyed and falls when it recovers. The measurement of TEWL was performed using AUM-3 manufactured by Forsion.

(2) Results The results are shown in Table 2. As is clear from Table 2, TEWL of the ceramide synthesis accelerator applied groups of Examples 1, 4 and 5 three days after application of retinoic acid was lower than that of the control, and the application of the ceramide synthesis accelerator of Example 1 resulted in retinoic acid. It was found that the damage of the skin barrier function was recovered.

Hereinafter, application examples of the ceramide synthesis accelerator of the present invention will be described. Application Examples 1 to 3 (skin cream) The ceramide synthesis accelerator of Example 1 was added to the composition (% by weight,
The same applies hereinafter) to prepare skin creams (Application Examples 1 to 3). (1) Composition

[0035]

[Table 3]

(2) Preparation Method The components (A) and (B) were each heated and dissolved at 80 ° C., then mixed, cooled with stirring, and cooled to 30 ° C. to prepare a skin cream.

Application Examples 4 to 6 (Lotion) The ceramide synthesis accelerator of Example 1 was blended with the composition shown in Table 4,
Lotions were prepared (Formulation Examples 4 to 6). (1) Composition

[0038]

[Table 4]

(2) Preparation method Each component was mixed and dissolved to prepare a lotion.

Application Examples 7 to 9 (Lipophilic cream) The ceramide synthesis accelerator of Example 1 was blended with each of the compositions shown in Table 5 to prepare creams (Formulation Examples 7 to 9). (1) Composition

[0041]

[Table 5]

(2) Preparation Method The components (A) and (B) were each heated and dissolved at 60 ° C., then mixed, cooled with stirring, and cooled to 30 ° C. to prepare a cream.

Application Examples 10 to 11 (Serum) The ceramide synthesis promoter of Example 1 was blended with each of the compositions shown in Table 6 to prepare serums (Formulation Examples 10 to 11). (1) Composition

[0044]

[Table 6]

(2) Preparation Method The components (A) and (B) were each heated and dissolved at 60 ° C., then mixed, cooled with stirring, and cooled to 30 ° C. to prepare a serum.

[0046]

As described above, the present invention is expected to improve rough skin and various skin diseases by activating ceramide synthesis of the epidermal cells themselves in the skin surface layer and improving the skin barrier function. It is clear that a ceramide synthesis accelerator having excellent stability can be provided.

Claims (1)

[Claims] 1. A ceramide synthesis comprising a bacterial culture having a ceramide synthesis promoting action as component A) and 1,3-butylene glycol as component B), wherein component A) is 3 to 6 times the weight of component B). Accelerator.