Oral compositions and topical skin preparations

Abstract

To reduce restrictions in the design of a neutrophil elastase inhibitor for use as a skin external preparation and an internal preparation.SOLUTION: A wrinkle and / or sag prevention and improvement oral composition includes variolaric acid.SELECTED DRAWING: Figure 3

Description

Technical Field

[0001] The present invention relates to an oral composition and a topical skin preparation for preventing and improving wrinkles and sagging.

Background Art

[0002] Elastin is a fibrous protein that generates the elasticity of tissues and is highly expressed in tissues that require elasticity, such as arteries, ligaments, and lungs. Elastase is a protease that degrades elastin, and there are eight elastase genes in humans. Neutrophils in the blood vessel adhere to the blood vessel inner wall using adhesion factors on the inner wall as a foothold, and then pass through between endothelial cells of the blood vessel and exit outside the blood vessel. As shown in Patent Documents 1 and 2, neutrophil elastase released by the neutrophils has been reported to degrade elastin in the dermis, damage the extracellular matrix, and contribute to the formation of wrinkles. In addition, it has been reported that when the skin is exposed to ultraviolet rays or physical stimuli such as repeated formation of wrinkles in the same part of the skin such as the outer corner of the eye when laughing, the amount of neutrophil elastase secreted increases in response to the inflammatory reaction. Therefore, it is considered that inhibiting the action of neutrophil elastase suppresses wrinkle formation. In addition, as a neutrophil elastase inhibitor, a preventive or improving agent for wrinkles and / or sagging containing a specific compound is known particularly by the general formula (1) in Patent Document 1. In addition, topical and oral preparations containing a neutrophil elastase inhibitor having an extract of Hydrastis canadensis as an active ingredient and expected to maintain a youthful skin condition are known from Patent Document 2.

Prior Art Documents

Patent Documents

[0003]

Patent Document 1

Patent Document 2

Non-Patent Documents

[0004] [Non-Patent Document 1] 2010 Dec;60(3):151-8. Neutrophil elastasecontributes to extracellular matrix damage induced by chronic low-dose UVirradiation in a hairless mouse photoaging model [Non-Patent Document 2] Journal of the Japanese Society for Fragrance and Cosmetics, Vol. 44, No. 1, 2020, pp. 13-19. [Overview of the project] [Problems that the invention aims to solve]

[0005] While prior neutrophil elastase inhibitors certainly exhibit some efficacy, their effectiveness is limited to specific compounds or plant extracts, resulting in a limited selection of alternative neutrophil elastase inhibitors. Expanding the range of neutrophil elastase inhibitor options was desired. Ultimately, this aims to reduce the limitations on the design of topical and oral formulations of neutrophil elastase inhibitors. [Means for solving the problem]

[0006] The main components of this invention are as follows: 1. An oral composition containing varioralic acid for the prevention and improvement of wrinkles and / or sagging. 2. An oral composition containing varioralic acid for inhibiting neutrophil elastase activity. 3. A topical skin preparation containing varioralic acid for the prevention and improvement of wrinkles and / or sagging. 4. A topical skin preparation containing varioralic acid for inhibiting neutrophil elastase activity. [Effects of the Invention]

[0007] Novel neutrophil elastase inhibitors can enhance the range of oral compositions and topical skin preparations that can prevent and suppress wrinkles and sagging by inhibiting the action of neutrophil elastase. [Brief explanation of the drawing]

[0008] [Figure 1] Structural formula of varioral acid. [Figure 2] This figure shows SDS electrophoresis results of elastin degradation inhibition evaluated by adding elastase and candidate inhibitors to proteins extracted from elastin-high-expression cells. [Figure 3] A figure showing the dependence of neutrophil elastase inhibitory activity on the concentration of varioralic acid. [Modes for carrying out the invention]

[0009] The improved oral composition and topical skin preparation of the present invention contain varietyolaric acid.

[0010] (Varioralic acid) The varioralic acid used in this invention is a compound having the structure shown in Figure 1, and is a compound found in certain lichens such as mosses. Varioral acid may be obtained by synthesis, or by extraction from specific plants or lichens containing varietyoral acid (e.g., Usnia) using any extraction method, or by purifying the extract to obtain a high-purity version. Furthermore, by incorporating varioralic acid into oral compositions or topical skin preparations, neutrophil elastase inhibitory activity in skin and tissues can be easily enhanced. In this way, inhibiting the activity of neutrophil elastase reduces the degree to which dermal elastin is broken down and the extracellular matrix is ​​damaged. As a result, it is possible to prevent the occurrence of wrinkles and sagging, improve existing wrinkles and sagging, and ultimately prevent skin aging and maintain skin surface firmness. The oral composition and topical skin preparation of the present invention can also prevent the occurrence of wrinkles and sagging, improve existing wrinkles and sagging, prevent skin aging, and maintain skin surface firmness.

[0011] (Oral composition) The oral composition in this invention is not particularly limited as long as it can contain varioralic acid. It can also be used as a composition for food and beverages, health foods, food additives, and pharmaceuticals. Alternatively, it can be made into a powder, tablet, or capsule using excipients commonly used in pharmaceuticals. Furthermore, as a food or beverage or health food, it may be in any form other than a liquid, such as an emulsion, paste, gel, powder, granules, pellets, sticks, or solid form.

[0012] Examples of such oral compositions include beverages such as coffee, tea, soft drinks, lactic acid bacteria drinks, fruit juice-free drinks, fruit juice drinks, and nutritional drinks; alcoholic beverages such as liqueurs and fruit wines; frozen desserts such as ice cream and sherbet; snacks; nutritional foods; desserts such as jelly, pudding, and yokan; confectionery such as cookies, cakes, chocolate, chewing gum, and manju; bread such as sweet buns and sliced ​​bread; candy such as ramune and tablets; meat products such as ham and sausages; fish products such as chikuwa and kamaboko; seafood products; and seasonings such as dressings, soy sauce, jam, and furikake. Thus, when used as an oral composition, it can be used in combination with antioxidants (such as vitamin C and vitamin E), sugars, sweeteners, thickeners and stabilizers, pH adjusters, etc., as a composition for food and beverages, health foods, food additives, and pharmaceuticals.

[0013] When varioralic acid is used in pharmaceutical or supplement formulations, it can be easily obtained by mixing and dispersing it directly or with a formulation-acceptable carrier using general means, and then processing it into the desired form. In this case, in addition to the varioralic acid used in the present invention, oily components such as vegetable oils and animal oils, vitamins, pH adjusters, preservatives, thickeners, pigments, fragrances, etc., which are commonly used in such forms, can be appropriately blended within a range that does not interfere with the effects of the present invention. The content of variolic acid of the present invention in these oral compositions is preferably 0.1 ppm or more, more preferably 1.0 ppm or more, still more preferably 5.0 ppm or more, and most preferably 10.0 ppm or more in the solid content of food and drink. Also, it is preferably 99.0% by mass or less, more preferably 10.0% by mass or less, still more preferably 5.0% by mass or less, and most preferably 3.0% by mass or less.

[0014] (External preparation for skin) Examples of the external preparation for skin of the present invention include lotion, emulsion, cream, oil, ointment, pack, lip, lipstick, foundation, eyeliner, blush, mascara, eyeshadow, manicure / pedicure, nail coating agent, nail coating remover, beard shaving agent, shampoo, conditioner, hair treatment, hair tonic, hair spray, hair cream, hair lotion, hair styling agent, hair growth agent, permanent solution, hair dyeing agent, hand soap, body soap, dentifrice, mouthwash, facial cleanser / soap, perfume, anti-odor and antiperspirant agent, ointment, poultice, etc.

[0015] When the external preparation for skin of the present invention is used as a cosmetic, it can be used as a preparation for cosmetics and quasi-drugs by using a base, additives, etc. usually used in cosmetics in combination. The form of the external preparation for skin of the present invention can be a solution, emulsion, polymer gel preparation such as lotion, emulsion, cream, gel, etc. Also, it may be a foamed preparation, multi-layered preparation, spray preparation, sheet or gel pack preparation impregnated in a non-woven fabric, etc. The external preparation for skin of the present invention can contain medicinal components such as a humectant, surfactant, thickener, anti-inflammatory agent, vitamins, antioxidant, blood circulation promoter, wound healing agent, antibacterial substance, skin activator, resident bacteria control agent, active oxygen scavenger, whitening agent, etc. as optional components according to the purpose. The content of variolic acid of the present invention in these external skin preparations is preferably 0.1 ppm or more, more preferably 1.0 ppm or more, still more preferably 5.0 ppm or more, and most preferably 10.0 ppm or more in the solid content of the external skin preparation. Further, it is preferably 99.0% by mass or less, more preferably 10.0% by mass or less, still more preferably 5.0% by mass or less, and most preferably 3.0% by mass or less.

[0016] The pharmacokinetic ingredients that may be included in the topical skin preparation of the present invention are not particularly limited as long as they have been conventionally used in pharmaceuticals, quasi-drugs, cosmetics, sanitary materials, etc., and are soluble or dispersible in water. Specifically, these include Angelica keiskei extract, avocado extract, Hydrangea macrophylla extract, Althaea officinalis extract, Arnica montana extract, apricot extract, apricot kernel extract, fennel extract, turmeric extract, oolong tea extract, Echinacea japonica leaf extract, Scutellaria baicalensis extract, Phellodendron amurense extract, barley extract, Nasturtium officinale extract, orange extract, dried seawater, hydrolyzed elastin, hydrolyzed wheat powder, hydrolyzed silk, chamomile extract, carrot extract, Artemisia capillaris extract, licorice extract, karcade extract, kiwi extract, and cinchona extract. Su, cucumber extract, guanosine, bamboo grass extract, walnut extract, grapefruit extract, clematis extract, yeast extract, burdock extract, comfrey extract, collagen, cranberry extract, bupleurum extract, umbilical cord extract, salvia extract, soapwort extract, bamboo grass extract, hawthorn extract, shiitake mushroom extract, rehmannia extract, lithospermum extract, linden extract, meadowsweet extract, calamus root extract, birch extract, horsetail extract, honeysuckle extract, ivy extract, celandine extract Crataegus monogyna extract, elderberry extract, yarrow extract, peppermint extract, mallow extract, Swertia japonica extract, jujube extract, thyme extract, clove extract, cogongrass extract, citrus peel extract, spruce extract, Houttuynia cordata extract, tomato extract, natto extract, carrot extract, wild rose extract, hibiscus extract, Ophiopogon japonicus extract, parsley extract, honey, parietaria extract, Isodon japonicus extract, bisabolol, coltsfoot extract, butterbur extract, Poria cocos Examples of extracts include leopard extract, butcher's broom extract, grape extract, propolis, loofah extract, peppermint extract, linden extract, hop extract, pine extract, horse chestnut extract, skunk cabbage extract, soapberry 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, and others.

[0017] Furthermore, it contains 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; anti-inflammatory agents such as ε-aminocaproic acid, glycyrrhizic acid, β-glycyrrhetinic acid, lysozyme chloride, guaiazulene, and hydrocortisone; vitamins such as vitamins A, B2, B6, D, E, calcium pantothenate, biotin, and nicotinamide; antioxidants such as tocopherol, carotenoids, flavonoids, tannins, lignans, and saponins; blood circulation promoters such as γ-oryzanol and vitamin E derivatives; wound healing agents such as retinol and retinol derivatives; biopolymers such as deoxyribonucleic acid, sodium chondroitin sulfate, collagen, elastin, chitin, chitosan, and hydrolyzed eggshell membrane; allantoin, diisopropylamine dichloroacetate, and 4-aminomethylcyclohexyl Active ingredients such as xancarboxylic acids; also include cepharanthine, capsicum tincture, hinokitiol, garlic iodide 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, ethinylestradiol, carpronium chloride, benzalkonium chloride, diphenhydramine hydrochloride, tacanal, camphor, salicylic acid, vanillyl nonyl acid, vanillyl nonanoate, piroctone olamine, glyceryl pentadecanoate, mononitroguaiacol, resorcinol, gamma-aminobutyric acid, benzethonium chloride, mexiletine hydrochloride, auxin, female hormones, cantharis tincture, cyclosporine, hydrocortisone, polyoxyethylene sorbitan monostearate, etc. [Examples]

[0018] The present invention will be described in detail below based on examples, but the present invention is not limited thereto.

[0019] <Elastin degradation inhibition test> 293FreeStyle cells were transfected with the pcDNA3.1 / Elastin-Flag-6His plasmid to obtain cells that exhibited stable expression. Lysate containing elastin (elastin protein extract) was collected using RIPA, neutrophil elastase and the test substance were added, and the cells were incubated at 37°C for 2 hours. The samples were subjected to SDS electrophoresis, and elastin protein was confirmed by Western blotting using the Flag antibody. Elastin degradation and inhibition of degradation were evaluated. The results are shown in Figure 2. Here, for the elastin protein extract solution described above, sample 1 (control) was prepared by adding only the buffer. Sample 2 was prepared by adding neutrophil elastase, sample 3 was prepared by adding neutrophil elastase and the elastase inhibitor sivelestat, and sample 4 was prepared by adding neutrophil elastase and varioralic acid. The concentrations used for each sample were neutrophil elastase 90 ng / 50 μL and varioralic acid 1 mM. As shown in Figure 2, in sample 2, which did not contain a neutrophil elastase inhibitor, elastin protein was degraded by neutrophil elastase, while in samples 1, 3, and 4, elastin protein remained undegraded. From these results, it was confirmed that, similar to sample 3, varioralic acid also possesses neutrophil elastase inhibitory activity in sample 4, which is an example in accordance with the present invention.

[0020] <Concentration-dependent effect of valioralic acid's neutrophil elastase inhibitory activity> Neutrophil elastase hydrolyzes the elastin-specific substrate Suc-Ala-Ala-Ala-pNA (Succinyl-L-alanyl-L-alanyl-L-alanine p-nitroanilide) to produce p-nitroaniline, which becomes detectable at 410 nm. Therefore, the amount of p-nitroaniline produced can be used as an indicator to evaluate the neutrophil elastase inhibitory effect of the material. Based on this method, we constructed a 96-well scale evaluation system and evaluated it using absorbance. [method] (1) Obtaining reagents Native human Neutrophil Elastase protein [Active] (ab91099) was obtained from Abcam. Suc-Ala-Ala-Ala-pNA(3071-V) was obtained from the Peptide Institute. The elastase inhibitor sivelestat (S7198) was obtained from Sigma. (2) Neutrophil elastase inhibition test Buffer (100mM HEPES pH 7.5, 500mM NaCl, 0.05% Tween20), varioralic acid (0.0001μM, 0.001μM, 0.01μM, 0.1μM, 1μM, 10μM, 100μM), and neutrophil elastase (final concentration 100ng / well) were applied to a 96-well transparent plate (IWAKI), and incubated at 37°C for 15 minutes. Suc-Ala-Ala-Ala-pNA, an elastin-specific substrate, was added at a final concentration of 12.5mM, and the reaction was further incubated at 37°C for 60 minutes. The absorbance at 410nm was measured, and the inhibition rate was calculated using the following formula. Neutrophil elastase inhibition rate (%) = 1 - (absorbance of test substance / absorbance of control) × 100 The results are shown in Figure 3. As shown in Figure 3, it can be confirmed that increasing the concentration of varioralic acid from 0.1 μM to 1 μM increases the neutrophil elastase inhibition rate. Furthermore, it can be seen that increasing the concentration of varioralic acid from 10 μM to 100 μM increases the neutrophil elastase inhibition rate to over 80%. This indicates that varioralic acid inhibits the degradation of Suc-Ala-Ala-Ala-pNA, an elastin-specific substrate. This suggests that varioralic acid inhibits elastin degradation by inhibiting the activity of neutrophil elastase. Therefore, by employing the present invention, a sufficiently high level of neutrophil elastase activity inhibition can be achieved, and consequently, wrinkles and / or sagging of the skin can be prevented or improved.

Claims

1. An oral composition containing varioralic acid for inhibiting neutrophil elastase activity.

2. A topical skin preparation containing varioralic acid for inhibiting neutrophil elastase activity.

Images