JP3435181B2 - External preparation for melanin production suppression - Google Patents

Description

Description: BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to the application of a yeast culture to an external preparation, and more particularly, to a specific sugar.
The present invention relates to an external preparation for inhibiting melanin production, which effectively utilizes a melanin production inhibitory effect and a moisturizing effect of a yeast culture solution or a culture extract inoculated with a specific bacterial species in a chemical solution . [0002] The field of utilization of yeast is wide, and fermentation products of yeast have been used in the food and pharmaceutical fields for a long time. In particular, the use of yeast cell extract is the mainstream, and it is known that yeast extract is used in cosmetics and the like utilizing the moisturizing effect and skin cosmetic effect of yeast extract. For example, Japanese Patent Publication No. 39-4 using a yeast cell itself as a pack
No. 899, JP-B-56-44 discloses a melanin production inhibitor comprising a mixture of an enzyme hydrolyzate having a placental composition and a yeast extract.
No. 046, Japanese Patent Publication No. 61-260009, which discloses a yeast extract containing 50 to 5000 PPM of RNA, and Japanese Patent Publication No. 61-171405, which discloses a cosmetic containing an organic solvent extract of brewer's yeast cells. Etc. are known. [0003] Many uses of yeast cell extracts are focused on their moisturizing properties, and some of them are intended to suppress the production of melanin by a synergistic effect with others. is there. Conventional methods for extracting useful substances from yeast cells have been difficult to mass-produce because cells that can be obtained only in small amounts are used. On the other hand, as an example using a yeast culture solution, Japanese Patent Publication No. Sho 6 (1995) discloses an anemia ameliorating composition containing a yeast culture solution or a yeast extract as an active ingredient.
No. 1-118322 and Japanese Patent Publication No. 64-11612, which disclose a sugar fermentation solution for cold waves, are only found, but their use as skin external preparations is not known. Accordingly, an object of the present invention is to focus not only on substances that enhance moisturizing properties but also on substances that exhibit the melanin production inhibitory effect required for cosmetics, An object of the present invention is to provide an external preparation having a melanin production inhibitory effect and a moisturizing effect by producing both of them and increasing their concentrations. SUMMARY OF THE INVENTION The present invention has been proposed to achieve the above-mentioned object, and a melanin is added to a yeast culture or a culture extract obtained by inoculating a specific saccharified solution with a specific bacterial species. The present invention has been completed based on the knowledge obtained by the present inventor that it has a production suppressing effect and a moisturizing effect. That is, according to the present invention, wheat is converted into a saccharifying enzyme.
And simultaneously adding α-amylase and glucoamylase
An external preparation for suppressing melanin production, characterized in that the obtained saccharified solution contains a yeast culture solution or a culture extract inoculated with Saccharomyces cerevisiae or Saccharomyces curlsbergensis as a bacterial species. DETAILED DESCRIPTION OF THE INVENTION The present inventors have conducted intensive studies using yeast of the genus Saccharomyces to produce a melanogenesis inhibitor and a moisturizing component in a culture solution.
6 cells), it was found that a culture solution of yeast of the genus Saccharomyces in a sugar solution, particularly a medium using wheat, strongly suppresses the production of melanin by pigment cells, and at the same time, has a strong moisturizing property. Thus, the present invention has been completed. In the present invention, the term "yeast culture solution" refers to a culture solution itself or a culture solution obtained by filtering the culture solution with a membrane filter or the like and removing the bacteria with a centrifugal separator or the like, or a solution that has been treated to increase the concentration of a melanin production inhibitor. The culture extract is obtained by concentrating this culture solution with a vacuum concentrator or the like.
The yeast Saccharomyces according to the present invention, whip
Saccharomyces cerevisiae
Or Saccharomyces Carlsbergensis (Saccharomy
ces carlsbergensis), among which Saccharomyces cerevisiae is more effective. In the present invention, the sugar solution, that is, the carbon source, is barley (eg, barley, wheat, rye, oat, etc.) or barley, which is heated and swelled , and then α-α
Amylase and glucoamylase added simultaneously
The is intended to be used in the medium, thereby increases the production of melanin production inhibitors by yeast. When the sugar solution does not sufficiently contain a nitrogen source, it is preferable to add peptone, a soybean protein hydrolyzate, an ammonium salt, an amino acid and the like in an appropriate amount. The melanin production inhibitory effect of the yeast culture solution in the present invention is sufficient even with a culture solution or a concentrated solution thereof, but the effect is further enhanced by addition of an organic solvent such as alcohols, salting out, an adsorbent, and fractionation by column chromatography. It is also possible.
The yeast culture solution or culture extract of the present invention obtained as described above has an excellent melanin production inhibitory effect and a moisturizing effect, and does not cause any trouble such as damage to the skin and is also excellent in safety. I have. The external preparation for inhibiting melanin production of the present invention is prepared by blending the above-mentioned active ingredient in a known dosage form which is acceptable for skin application. Usually, it is advisable to mix the culture solution or culture extract in the preparation at about 0.05 to 50% by weight, preferably about 0.1 to 10% by weight. Further, in addition to the active ingredient of the present invention, various known active ingredients usually used, for example, vitamin E,
Vitamin E such as vitamin E nicotinate, nicotinic acid, nicotinamide, and benzyl nicotinate; peripheral vasodilators such as cepharanthin, ginger tincture, and capsicum tincture; refreshing agents such as camphor and menthol; hinokitiol; benzalkonium chloride; undecylene Antibacterial agents such as acid, anti-inflammatory agents such as lysozyme chloride, glycyrrhizin, allantoin, whitening agents such as kojic acid and its derivatives, ascorbic acid and its derivatives, arbutin liquiritin and its derivatives, assembly extracts, garlic extract, carrot extract, and gongon extract , Rosemary extract, aloe extract, luffa extract, ginkgo extract,
Various extracts derived from animals, plants, and microorganisms, such as elder extract, placenta extract, liver extract, and lactic acid bacteria culture extract, can be freely added and used. The known dosage form of the external preparation of the present invention means any dosage form that can be used externally, such as cataplasms, plasters, pastes, creams, ointments, aerosols, emulsions, lotions, emulsions, and essences. , Packs, gels,
Powder, foundation, sun care, bath salt,
Skin application agents such as soaps can be exemplified. In addition, the above-mentioned external preparations include, in addition to known active ingredients, surfactants, base components such as oils and fats, and a known moisturizing agent, if necessary, as long as the effect as the external preparation for suppressing melanin production is not impaired. Various additives such as agents, thickeners, preservatives, antioxidants, ultraviolet absorbers, scattering agents, chelating agents, pH adjusters, fragrances and coloring agents can be used in combination. Examples of humectants include glycerin, propylene glycol, 1,3-butylene glycol, sorbitol, mannitol, polyethylene glycol,
Polyhydric alcohols such as dipropylene glycol, amino acids, NMF components such as sodium lactate and sodium pyrrolidone carboxylate, hyaluronic acid, collagen, elastin, chondroitin sulfate, dermatan sulfate, fibronectin, ceramides, heparin-like substances, chitosan, etc. Examples include water-soluble polymer substances. Examples of the thickener include sodium alginate, xanthan gum, aluminum silicate, quince seed extract, tragacanth gum, natural polymer substances such as starch, methylcellulose, hydroxyethylcellulose, carboxymethylcellulose, soluble starch, cationized cellulose and the like. And a synthetic polymer such as carboxyvinyl polymer and polyvinyl alcohol. Examples of preservatives include benzoate, salicylate, sorbate, dehydroacetate, paraoxybenzoate, 2,4,4'-trichloro-2'-hydroxydiphenyl ether, 3,4,4'-trichloroform. Examples include carbanilide, benzalkonium chloride, hinokitiol, resorcinol, ethanol and the like. Examples of the antioxidant include dibutylhydroxytoluene, butylhydroxyanisole, propyl gallate, ascorbic acid and the like. Examples of the ultraviolet absorber include 4-methoxybenzophenone, octyldimethylparaaminobenzoate, ethylhexylparamethoxycinnamate,
Examples include titanium oxide, kaolin, and talc. Further, examples of the chelating agent include ethylenediaminetetraacetate, pyrophosphate, hexametaphosphate, citrate, tartaric acid, gluconic acid, and the like. ,
Examples thereof include potassium hydrogen phosphate. EXAMPLES The following are production examples of the present invention, test examples and prescription examples for explaining the effects thereof, but they do not limit the present invention in any way. <Production Example 1> 1 kg of barley (barley), 400 g of glucose, 80 g of hydrolyzate of soybean protein and 1 g of purified water
After adding 8.52 liters and adjusting the pH to 5, the temperature was adjusted to 120 ° C.
After heating for 15 minutes and cooling to 30 ° C, equal amounts of α-amylase and glucoamylase are aseptically added in small amounts, and inoculum of yeast (Saccharomyces cerevisiae) was inoculated thereto. The cells were cultured for about 6 days until the amount became 0.1 to 0.5%. The culture was sterilized with a membrane filter (0.25 µ) and then concentrated 5 times to obtain about 4 kg of the product of the present invention. <Production Example 2> Purified water was added to 1.4 kg of wheat.
Then, the mixture is boiled and swollen, cooled to 40 ° C., and α-amylase and glucoamylase are added in small amounts in equal amounts to saccharify for about 2 hours. 100 g of peptone and 6.5 liters of purified water were added to the saccharified solution, the pH was adjusted to 5, and the mixture was sterilized by heating at 120 ° C. for 15 minutes, cooled, and then subjected to yeast (Saccharomyces cerevisiae).
visiae), and cultured at 30 ° C. for about 6 days until the residual amount of glucose in the culture solution became 0.1 to 0.5%. The culture was sterilized by centrifugation and then concentrated 10 times to obtain about 2 kg of the product of the present invention. <Production Example 3> To 1 kg of steamed barley and 40 g of peptone, 18.96 liters of purified water was added, sterilized by heating at 120 ° C. for 15 minutes, and cooled, followed by yeast (Saccharomyces cereal).
visiae) was inoculated thereto and cultured at 30 ° C. for 7 days. The culture was centrifuged to remove bacteria, and then concentrated 5 times to obtain about 4 kg of the product of the present invention. <Production Example 4> 1 kg of barley, 40 glucose
0 g and 200 g of peptone in 0.1 M phosphate buffer (p
H5) 18.6 kg was added, the mixture was sterilized by heating at 120 ° C. for 15 minutes, cooled, and then inoculated with yeast (Saccharomyces carlsbergensis), and cultured at 32 ° C. for about 8 days. The culture was centrifuged to remove bacteria, and then concentrated 10-fold with a vacuum concentrator to obtain about 2 kg of the product of the present invention. <Production Example 5> 18.6 liters of purified water was added to 1.2 kg of steamed barley and 200 g of broth, the pH was adjusted to 5, and the mixture was sterilized by heating at 120 ° C for 10 minutes. And saccharified. Yeast (Saccha)
romyces cerevisiae) and inoculated at 30 ° C. for 8 days. The culture was sterilized with a membrane filter and then concentrated 10-fold with a vacuum concentrator to obtain about 2 kg of the product of the present invention. <Production Example 6> 800 g of wheat, 400 g of corn starch, and 18.8 liters of water were boiled and swollen, and then heated to 40 g.
While maintaining at ℃, α-amylase and glucoamylase were added in small amounts in equal amounts, and saccharified for about 2 hours. This saccharified solution was adjusted to pH 5, sterilized by heating at 120 ° C. for 15 minutes, cooled, and inoculated with a yeast (Saccharomyces cerevisiae) inoculum. At 30 ° C., the amount of glucose in the culture solution was reduced to 0.5% or less. Cultured for 8 days. After removing the bacteria by centrifugation of the culture solution, 6% of synthetic resin Sepabeads SP-205 (manufactured by Mitsubishi Kasei Kogyo Co., Ltd.) was added, followed by stirring for 1 hour, and the synthetic resin was filtered off. The filtrate was concentrated to one-tenth volume with a vacuum concentrator to obtain about 2 kg of the product of the present invention. <Experimental example 1> Mouse melanoma B16 fine
Test method for whitening using cells Prepare and dissolve the sample in MEM (Eagle's Minimum Essential Medium) so that the final concentration is as shown in Table 1.
The solution was filtered through a sterilizing filter having a pore size of 0.45 μm. MEM
The insoluble sample was dissolved in 100 μl of ethanol,
Added to MEM. Two plastic Petri dishes (Falc
1 × 10 ⁵ / ml B16 suspended in 8 ml of MEM, 1 ml of FBS (fetal bovine serum) and 1 ml of MEM, each of which was dissolved and filtered and sterilized.
The cells were added, and the medium was replaced three days after the start of the culture.
The cells were cultured at 37 ° C. for 5 days under the conditions of 5% CO ₂ and 95% air. After culturing, collect the cells grown on the bottom of the Petri dish,
Suspend in Phosphate buffered saline (PBS) and add 2
Centrifugation was performed at 000 rpm for 3 minutes, and the degree of blackening of the obtained cell pellet was visually evaluated. In Table 1, +-
Indicates the following evaluation. ±: shows the same degree of blackening as the non-added group. +: Shows a slightly lower degree of blackening than the non-added group. ++: The degree of blackening is clearly lower than that of the non-added group. +++: Degree of blackening slightly observed. ++++: White to gray, not recognized as black. ++++++: white color <Experimental example 2> Guinea pig ultraviolet pigmentation
Inhibitory effect The effect of improving pigmentation was examined using tan guinea pigs. Table 2 shows the results. [Test Method] Using the skin of the back of ten tan guinea pigs, the back hair of the guinea pig was shaved with a clipper and further shaved with an electric razor. The back of this guinea pig,
UV-B (three SE lamps, 140 mJ) covered with aluminum foil with holes of 9 squares (2.0 x 2.0 cm) at four places
/ Cm ² ) once a day for 90 seconds, three times every three days.
"Wheat fermented liquor" from the start of irradiation (Production Examples 1 to 3 )
Was applied three times a day for 20 consecutive days. 1 after start of application
The judgment was made between the 3rd and 20th days. The degree of blackening of the skin color was visually determined according to the following criteria. Evaluation criteria Excellent: no pigmentation was observed. Effective: slight pigmentation is observed. Moderately effective: moderate pigmentation is observed. Invalid: No difference from control site. Reversal: stronger pigmentation than control is observed. [0031] <Experimental example 3> Impedance meter
According moisture Test [Test method] frequency impedance meter (IBS Co.: MODEL IB-335) used to examine the moisture of 5% yeast culture of purified water as a control (preparation 1). For comparison with other humectants, a 0.5% solution of sodium hyaluronate having high moisturizing properties was similarly examined as a control. [Measurement method] Apply sample to human forearm flexion side (2 cm × 2
cm), and after 30 seconds, quickly and gently wipe with gauze.
Every 0 seconds, the skin conductance (conductance) is increased by 1 over time.
Measurement was performed up to 0 minutes (measurement conditions: measurement room temperature 20 ° C., humidity 60%, number of measurements n = 10). [Test Results] The results are shown in FIG. The results in FIG. 1 show that the higher the conductivity, the higher the moisturizing property, and it was confirmed that the yeast culture solution of the present invention was excellent as a moisturizing agent. The following are formula examples. In the prescription examples,
“Appropriate amount” means an amount that makes the total amount of the formulation 100% by weight. <Prescription Example 1> Cream (% by weight) Polyethylene glycol monostearate (40E.0.) 2.00 2. 2. Self-emulsifying glyceryl monostearate 5.00 Stearic acid 5.00 4. Behenyl alcohol 1.005. Liquid paraffin 10.00 6. Glyceryl trioctanoate 10.00 7. 7. Fermented yeast liquor (obtained in Production Example 1) 4.00 Paraoxybenzoate 0.20 9.1 1,3-butylene glycol 5.00 10. Disodium edetate 0.01 11. Purified water Proper quantity production method A. Heat and dissolve 1 to 7. B. Heat and dissolve 8-12. C. A. Add B to A, emulsify, stir and cool. After cooling, C is filled into a container, and is used as a product after inspection. Rub face and appropriate dosage and dosage. <Formulation Example 2> Emulsion (% by weight) Polyoxyethylene sorbitan monostearate (20E.0.) 2.00 2. 2. Polyoxyethylene sorbite tetraoleate (60E.0.) 0.50 3. Lipophilic monostearin type glycerin 1.00 Stearic acid 0.50 5. Behenyl alcohol 0.50 6. Avocado oil 4.00 7. Glyceryl trioxitanate 4.00 8. 8. Yeast fermentation liquor (obtained in Production Example 3) 3.00 Paraoxybenzoate 0.20 10.1,3 Butylene glycol 5.00 11. Xanthan gum 0.14 12. Disodium edetate 0.01 13. Purified water Proper quantity production method A. Heat and dissolve 1-8. B. Heat and dissolve 9-14. C. A. Add B to A, emulsify, stir and cool. After cooling, C is filled into a container, and is used as a product after inspection. Rub face and appropriate dosage and dosage. <Formulation Example 3> Lotion agent (% by weight) Polyoxyethylene hydrogenated castor oil (60E.0.) 8.00 2. Ethanol 15.00 3. 3. Yeast fermentation liquid (obtained in Production Example 6) 5.00 4. Paraoxybenzoate 0.10 5. citric acid 0.10 7. Sodium citrate 0.30 7.1, 3-butylene glycol 4.00 Disodium edetate 0.019. Purified water Proper quantity production method A. 1 to 10 are uniformly stirred and dissolved. B. A is filled in a container, and is used as a product after inspection. Rub face and appropriate dosage and dosage. <Formulation Example 4> Cream pack (% by weight) Polyethylene glycol monostearate (40 E.0.) 2.00 2. 2. Self-emulsifying glyceryl monostearate 5.00 Stearic acid 5.00 4. Behenyl alcohol 0.50 5. Squalane 15.00 6. 6. Cetyl octanoate 5.00 7. Yeast fermented liquid (obtained in Production Example 2) 5.00 Paraoxybenzoate 0.20 9.1 1,3-butylene glycol 5.00 10. Disodium edetate 0.01 11. Purified water Proper quantity production method A. Heat and dissolve 1 to 7. B. Heat and dissolve 8-12. C. A. Add B to A, emulsify, stir and cool. After cooling, C is filled into a container, and is used as a product after inspection. Rub face and appropriate dosage and dosage. <Formulation Example 5> Ointment (% by weight) 1. Polyoxyethylene sorbitan monostearate (20E.0.) 1.00 2. Polyoxyethylene sorbite tetraoleate (60E.0.) 1.50 Self-emulsifying glyceryl monostearate 1.50 4. Salami beeswax 2.00 5. Paraffin 2.00 6. Stearic acid 3.00 7. 7. Behenyl alcohol 3.00 Liquid paraffin 5.00 9. Avocado oil 12.00 10. Vitamin E 12.00 11. 11. Yeast fermentation liquor (obtained in Production Example 4) 10.00 Paraoxybenzoic acid ester 0.20 13.1,3 Butylene glycol 5.00 14. Purified water qs Production method A. Heat and dissolve 1-11. B. Heat and dissolve 12-15. C. A. Add B to A, emulsify, stir and cool. After cooling, C is filled into a container, and is used as a product after inspection. Rub face and appropriate dosage and dosage. <Formulation Example 6> Emulsion (% by weight) 1. Polyoxyethylene sorbitan monostearate (20E.0.) 1.00 2. Polyoxyethylene sorbite tetraoleate (60E.0.) 0.50 3. Lipophilic glyceryl monostearate 1.00 Stearic acid 0.50 5. Behenyl alcohol 0.50 6. Avocado oil 4.00 7. 7. Glyceryl trioctanoate 4.00 8. Yeast fermentation liquor (obtained in Production Example 5) 3.00 Paraoxybenzoate 0.20 10.1,3 Butylene glycol 5.00 11. Xanthan gum 0.14 12. Disodium edetate 0.01 13. Purified water Proper quantity production method A. Heat and dissolve 1-8. B. Heat and dissolve 9-14. C. A. Add B to A, emulsify, stir and cool. After cooling, C is filled into a container, and is used as a product after inspection. Rub face and appropriate dosage and dosage. According to the present invention, a saccharifying enzyme specific to wheat is provided.
Inoculate a specific bacterial species into the saccharified solution obtained by adding
An external preparation that is remarkably excellent in a melanin production inhibitory effect comprising a yeast culture solution or a yeast extract provided is provided. It is preferably used.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a graph showing the electric power of a yeast culture solution of the sugar solution of the present invention in comparison with the electric power of sodium hyaluronate.

──────────────────────────────────────────────────の Continuation of front page (51) Int.Cl. ⁷ identification code FI A61K 38/00 A61P 17/00 A61P 17/00 A61K 37/02 (58) Field surveyed (Int.Cl. ⁷ , DB name) A61K 7/00-7/50 C12N 1/16-1/19 JICST file (JOIS)

Claims (1)

(57) [Claims 1] α-Amirror as a saccharifying enzyme in barley
An external preparation for suppressing melanin production, comprising a yeast culture solution or a culture extract inoculated with Saccharomyces cerevisiae or Saccharomyces curlsbergensis as a bacterial species to a saccharified solution obtained by simultaneously adding zeolites and glucoamylase .