JP7421695B2 - Melanin production inhibitors and skin whitening agents or skin external preparations containing the same - Google Patents

Description

The present invention relates to a melanin production inhibitor containing as an active ingredient one or more selected from spermidine, spermine, acetylspermidine, diacetylspermidine, and acetylspermine.

Due to air pollution and the destruction of the ozone layer, the amount of ultraviolet rays reaching the epidermis tends to increase year by year, and as a result, skin problems such as dark spots, freckles, and dark skin due to ultraviolet rays are becoming more common. When exposed to ultraviolet rays, tyrosine present in the skin is oxidized by the action of the tyrosinase enzyme, producing melanin pigment. Excessive production can cause skin problems such as age spots, freckles, and dark skin. As a method for suppressing the production of melanin pigment and preventing age spots, freckles, and dark skin, there have been conventional skin preparations containing arbutin (Patent Document 1: JP-A-60-16906), L-ascorbic acid, and L-ascorbic acid. A skin cosmetic containing a glucose glycoside of L-ascorbic acid, which is a derivative, has been proposed (Patent Document 2: JP-A-4-182412). Cosmetics containing arbutin (Patent Document 3: JP-A-2009-67691) and whitening agents containing kojic acid may not have sufficient whitening effects, or may contain ingredients that exhibit whitening effects at concentrations that are acceptable for whitening effects. When mixed with other substances, safety problems may arise.

In addition, polyamines such as spermidine and spermine, and acetylated polyamines such as acetylspermidine and acetylspermine are known to be markers of various cancers in blood and urine (Non-Patent Document 1: Soda K (2011)), melanin production suppressing effect, and whitening effect are not known.

Japanese Unexamined Patent Publication No. 16906/1986 Japanese Patent Application Publication No. 4-182412 JP2009-67691A International Publication No. 2011/081024

Soda K, “The mechanisms by which polyamines accelerate tumor spread”, 2011, Journal of Experimental & Clinical Cancer Research ,2011,30,95

An object of the present invention is to provide a novel melanin production inhibitor.

The main configuration of the present invention is as follows.
1. A melanin production inhibitor containing one or more selected from spermidine, spermine, acetylspermidine, diacetylspermidine, and acetylspermine as an active ingredient.
2.1. A whitening agent containing the melanin production inhibitor described in .
3.1. A skin external preparation containing the melanin production inhibitor described in .

Spermidine, spermine, acetylspermidine, diacetylspermidine, and acetylspermine used in the present invention have an effect of suppressing melanin production and have a whitening effect. It is possible to provide a skin whitening agent and a skin external preparation containing one or more selected from spermidine, spermine, acetylspermidine, diacetylspermidine, and acetylspermine as an active ingredient.

The structural formulas of spermidine and spermine are shown below (1) and (2), respectively.

The present invention provides a melanin production inhibitor containing as an active ingredient one or more selected from the polyamine compounds spermidine and spermine, and their acetyl derivatives acetylspermidine, diacetylspermidine, and acetylspermine, and a melanin production inhibitor using the same. Concerning skin whitening agents and external skin preparations.

The concentrations of spermidine, spermine, acetylspermidine, diacetylspermidine, and acetylspermine in the melanin production inhibitor of the present invention are not particularly limited as long as they are within a range that can suppress melanin production, but each concentration is 0.01 μg/mL or more and 200 μg /mL or less is preferable.

When the melanin production inhibitor of the present invention is used to exhibit a whitening effect, it can be mixed with bases, additives, etc. for various whitening agents to form the whitening agent of the present invention.

The skin whitening agent of the present invention includes cosmetics, foods, and pharmaceuticals, and when used as a cosmetic, it can be manufactured according to commonly used formulation methods, including oils and fats such as vegetable oil, high-quality Fatty acids, higher alcohols, silicones, anionic surfactants, cationic surfactants, amphoteric surfactants, nonionic surfactants, preservatives, sugars, sequestering agents, polymers such as water-soluble polymers, thickeners It can contain agents, powder components, ultraviolet absorbers, ultraviolet blockers, humectants such as hyaluronic acid, perfumes, pH adjusters, and the like. It is also possible to contain other medicinal and physiologically active ingredients such as vitamins, skin activators, blood circulation promoters, resident bacteria control agents, active oxygen scavengers, anti-inflammatory agents, whitening agents, and bactericidal agents.

When using the skin whitening agent of the present invention as a food, it may be incorporated into the desired food. For example, after adding suitable auxiliary agents such as starch, lactose, maltose, vegetable oil powder, cocoa butter powder, stearic acid, etc., it is possible to form a form suitable for oral intake, such as granules, granules, etc. using conventional means. It can be formed into tablets, capsules, pastes, etc. to produce health foods, health-promoting foods, functional foods, etc. It may also be added to various foods, such as processed meat foods such as ham and sausage, processed seafood foods such as kamaboko and chikuwa, bread, confectionery, butter, powdered milk, and fermented dairy products. It may be used by adding it to beverages such as drinks. When the skin whitening agent of the present invention is used as a pharmaceutical, it is mixed with a solid or liquid non-toxic pharmaceutical carrier suitable for administration methods such as oral administration, transdermal administration, intrarectal administration, and injection, and is then mixed with a conventional pharmaceutical preparation. It can be administered in the form of Examples of such preparations include solid preparations such as tablets, granules, powders, and capsules, liquid preparations such as solutions, suspensions, and emulsions, and lyophilized preparations. It can be prepared by conventional means.

The external skin preparation of the present invention includes cosmetics, quasi-drugs, and pharmaceuticals, and can be in the form of solutions, emulsions, or polymer gels, such as lotions, emulsions, creams, and gels. can. It may also be a foam preparation, a multilayer preparation, a spray preparation, a sheet impregnated with nonwoven fabric, or a gel pack preparation.

The external skin preparation of the present invention may include optional ingredients such as moisturizers, surfactants, thickeners, anti-inflammatory agents, vitamins, antioxidants, blood circulation promoters, wound healing agents, antibacterial substances, etc., depending on the purpose. Medicinal ingredients such as skin activators, resident bacteria control agents, active oxygen scavengers, and whitening agents can be included.

The medicinal ingredients are not particularly limited as long as they are conventionally used in pharmaceuticals, quasi-drugs, cosmetics, sanitary materials, etc. and can be dissolved or dispersed in water. Specifically, the extracts include Ashitaba extract, avocado extract, amacha extract, althea extract, arnica extract, apricot extract, apricot kernel extract, fennel extract, turmeric extract, oolong tea extract, scutellariae leaf extract, scutellariae extract, scutellariae extract, barley extract, and Dutch extract. Mustard extract, orange extract, dried seawater, hydrolyzed elastin, hydrolyzed wheat powder, hydrolyzed silk, chamomilla extract, carot extract, mugwort extract, licorice extract, karkade extract, kiwi extract, cinchona extract, cucumber extract, guanosine, blackberry extract , walnut extract, grapefruit extract, clematis extract, yeast extract, burdock extract, comfrey extract, collagen, lingonberry extract, psycho extract, rhinoceros extract, salvia extract, soapwort extract, bamboo grass extract, hawthorn extract, shiitake extract, rhododendron extract, shikon extract , linden extract, meadowsweet extract, calamus root extract, birch extract, horsetail extract, honeysuckle extract, Japanese ivy extract, hawthorn extract, Japanese elderberry extract, yarrow extract, mentha extract, mallow extract, Japanese commonweed extract, turmeric extract, thyme Extract, Clove extract, Chigaya extract, Chimpi extract, Spruce extract, Heutyami extract, Tomato extract, Natto extract, Carrot extract, Novara extract, Hibiscus extract, Bacteria extract, Parsley extract, Honey, Parietaria extract, Hikiokoshi extract, Bisabolol, Coltsfoot extract , butterbur extract, bud extract, butcher bloom extract, grape extract, propolis, loofah extract, peppermint extract, bodega extract, hop extract, pine extract, horse chestnut extract, skunk cabbage extract, sapinum extract, peach extract, cornflower extract, eucalyptus extract, yuzu extract, mugwort extract, lavender extract, apple extract, lettuce extract, lemon extract, astragalus extract, rose extract, rosemary extract, Roman chamomile extract, royal jelly extract, etc.

In addition, moisturizing agents such as amino acids, urea, sodium pyrrolidone carboxylate, betaine, whey, and trimethylglycine; oily components such as sphingolipids, ceramides, cholesterol, cholesterol derivatives, and phospholipids; ε-aminocaproic acid, glycyrrhizic acid, and β-glycyrrhetinic acid. Anti-inflammatory agents such as acids, lysozyme chloride, guaiazulene, and hydrocortisone; Vitamins such as vitamins A, B2, B6, D, and E, calcium pantothenate, biotin, and nicotinamide; tocopherols, carotenoids, flavonoids, tannins, lignans, and saponins. Antioxidants such as; blood circulation promoters such as γ-oryzanol and vitamin E derivatives; wound healing agents such as retinol and retinol derivatives; deoxyribonucleic acid, sodium chondroitin sulfate, collagen, elastin, chitin, chitosan, hydrolyzed eggshell membrane, etc. Biopolymer; active ingredients such as allantoin, diisopropylamine dichloroacetate, 4-aminomethylcyclohexanecarboxylic acid; cephalanthine, capsicum tincture, hinokitiol, iodized garlic extract, pyridoxine hydrochloride, dl-α-tocopherol, dl-α-tocopherol acetate , nicotinic acid, nicotinic acid derivatives, D-pantothenyl alcohol, acetyl pantothenyl ethyl ether, biotin, allantoin, isopropylmethylphenol, estradiol, ethinyl estradiol, carpronium chloride, benzalkonium chloride, diphenhydramine hydrochloride, takanal, camphor, salicylic acid, Nonylic acid vanillylamide, nonanoic acid vanillylamide, piroctone olamine, glyceryl pentadecanoate, mononitroguaiacol, resorcinol, γ-aminobutyric acid, benzethonium chloride, mexiletine hydrochloride, auxin, female hormone, cantharis tincture, cyclosporine, hydrocortisone, monostearic acid Also included are polyoxyethylene sorbitan and the like.

EXAMPLES Hereinafter, the present invention will be described in detail based on Examples, but the present invention is not limited thereto.
<Melanin production suppression test>
The inhibitory effect on melanin production was confirmed using B16 melanoma cells.
Putrescine dihydrochloride; (Nacalai Tesque, product code 29428-01)
Spermidine trihydrochloride; (Nacalai Tesque, product code 32110-41)
Spermine tetrahydrochloride; (Nacalai Tesque, product code 32113-11)
N8-acetylspermidine dihydrochloride; (Sigma-Aldrich, product code A3658)
N1,N8-Diacetylspermidine; (Toronto Research Chemicals Inc., product code D367000)
N1-acetylspermine trihydrochloride; (Sigma-Aldrich, product code 01467)
Kojic acid; (Fujifilm Wako Pure Chemical Industries, product code 118-00491)
Synthetic melanin; (Sigma-Aldrich, product code M8631)

(Test method)
After culturing B16 melanoma cells in Dulbecco's MEM medium containing 10% FBS (fetal bovine serum), the cells were collected by trypsin treatment. The collected cells were diluted with Dulbecco's MEM medium containing 10% FBS and 1 mmol/L theophylline to a concentration of 24.0×10 ⁴ cells/mL, and then seeded at 300 μL per well on a 48-well plate and cultured for 6 hours.
After the culture was completed, 300 μL of the test sample at a predetermined concentration dissolved in Dulbecco's MEM medium containing 10% FBS and 1 mmol/L theophylline was added to each well, and cultured for 4 days. After the culture was completed, the medium was removed from each well, 200 μL of a 2 mol/L NaOH solution was added, the cells were disrupted using an ultrasonic disruptor, and the absorbance at a wavelength of 475 nm was measured. From the measured absorbance value, the amount of melanin was calculated based on a calibration curve created using synthetic melanin. In addition, to measure cell viability, after culturing in the same manner, the cells were washed with 400 μL of PBS(-), and 200 μL of Neutral Red dissolved in Dulbecco's MEM containing 10% FBS at a final concentration of 0.05 mg/mL was added to each well. did. After culturing for 2.5 hours, the neutral red solution was discarded, and 200 μL of ethanol/acetic acid solution (ethanol:acetic acid:water=50:1:49) was added to each well to extract the pigment. After extraction, absorbance at a wavelength of 540 nm was measured. As a blank control, cells cultured only in Dulbecco's MEM medium containing 10% FBS and 1 mmol/L theophylline were tested in the same manner. The test was conducted with each sample n=3.

The melanin production inhibition rate was calculated using the following formula. The average values are shown in Table 1.
Melanin production suppression rate (%) = (1-(B/D)/(A/C)) x 100
A: Amount of melanin without the addition of the test sample B: Amount of melanin with the addition of the test sample C: Absorbance at 540 nm without the addition of the test sample D: Absorbance at 540 nm with the addition of the test sample Note: (D/C) x 100 (%) is the cell survival rate, and the melanin production suppression rate was not calculated for samples with a cell survival rate of less than 80%.

(result)
Spermidine, spermine, acetylspermidine, diacetylspermidine, and acetylspermine were confirmed to have an inhibitory effect on melanin production. No inhibitory effect on melanin production was confirmed for putrescine.
Furthermore, as a result of a similar test using kojic acid as a positive control, a melanin production inhibitory effect of 44.7±1.6% was confirmed at 200 μg/mL.
That is, it was confirmed that spermidine, spermine, acetylspermidine, and acetylspermine have an equivalent or higher melanin production inhibitory effect at lower concentrations than kojic acid, which is known to have a melanin production inhibitory effect.

(Prescription example)
(lotion)
1. Purified water remainder 2. Glycerin 5.0%
3. Diglycerin 7.0%
4.1,3-butylene glycol 5.0%
5. Betaine 1.0%
6. Spermine 0.001%
7. Phenoxyethanol 0.3%
1 to 7 were stirred and mixed to obtain a lotion.

(Beauty gel)
1. Purified water remainder 2. Dipropylene glycol 6.0%
3. PEG-75 0.5%
4. Raffinose 2.0%
5. Phenoxyethanol 0.2%
6. Pentylene glycol 1.0%
7. Xanthan gum 0.1%
8. Squalane 3.0%
9. SIMULGEL NS 2.5%
10. Spermidine 0.0001%
11. Ethanol 2.0%
Disperse 7 in 6 (dispersion). Mix 8 and 9 (mixture). The dispersion was gradually added to 1 to 5 which had been dissolved by heating, and the mixture was further added thereto, followed by stirring with a homomixer. After cooling, add 10 and 11, and stir again using a homomixer.
*SIMULGEL NS (manufactured by SEPPIC; mixture of (hydroxyethyl acrylate/acryloyl dimethyl taurine Na) copolymer, squalane, polysorbate 60, water, sorbitan isostearate)

(emulsion)
1. Olive oil 5.0%
2. Jojoba oil 1.0%
3. Dimethicone 0.5%
4. Polyglyceryl stearate-10 1.0%
5. Glycerin 7.0%
6. Dipropylene glycol 6.0%
7. Phenoxyethanol 0.5%
8.1,3-butylene glycol 3.0%
9. Xanthan gum 1.0%
10. Carboxyvinyl polymer 0.1%
11. Potassium hydroxide 0.02%
12. Acetyl spermine 0.002%
13. Disperse 9 in purified water Remaining 8 (dispersion). The dispersion is added to 10 dissolved in 13 by stirring, and then 5 to 7 are added and dissolved by heating (dissolved material). Dissolve 1 to 4 by heating, gradually add the dissolved material, and stir with a homomixer. Next, 11 dissolved in a portion of 13 is added, and after cooling, 12 dissolved in a portion of 13 is added, and the mixture is again stirred with a homomixer.

(Sheet beauty serum)
1. Purified water remaining 2.1,3-butylene glycol 5.0%
3. Glycerin 3.0%
4. Betaine 1.5%
5. Phenoxyethanol 0.1%
6. Carboxyvinyl polymer 0.1%
7. Potassium hydroxide 0.02%
8. Hyaluronate Na 0.001%
9. Acetyl spermidine 0.0005%
After stirring and dissolving 6 in 1, 2 to 5, 8, and 9 are added. After confirming dissolution, 7, which was partially dissolved in 1, is added and stirred again using a homomixer.

Claims (2)

A melanin production inhibitor containing as an active ingredient one or more selected from acetylspermidine, diacetylspermidine, and acetylspermine (excluding those containing acetylspermine and polyamine oxidase). A whitening agent containing the melanin production inhibitor according to claim 1.