JP4675806B2 - Annexin activator - Google Patents

Description

The present invention is an invention in the technical field relating to the prevention of skin aging and the prevention of skin roughness and pore conspicuousness due to keratinization, and the improvement thereof. The sodium salt and potassium salt of the compound represented by the chemical formula (1) Annexin activator.

  Skin damage caused by solar ultraviolet radiation, air pollutants, or many other causes can be divided into acute and chronic reactions. For example, inflammation is one of the typical skin disorders in the acute phase, and keratinization occurs in the epidermis, contributing to conspicuous rough skin and pores. In order to prevent and ameliorate such skin disorders, it is important to suppress inflammation, and treatment using a substance having an inhibitory action on the release of inflammatory chemical mediators such as histamine is generally used. However, such treatment is not always effective, and a fundamental solution for inflammation is desired.

  On the other hand, skin aging can be mentioned as a skin disorder in the chronic phase. Wrinkles, which are one type of skin aging, are aging changes accelerated by solar ultraviolet rays, and are caused by a decrease in skin elasticity due to a decrease and degeneration of extracellular matrix components such as collagen. In the dermis of skin exposed to such ultraviolet rays, protease with high resolution occurs using extracellular matrix components as a substrate, so fundamentally controlling the activation of degrading enzymes prevents skin aging It is extremely important.

In order to analyze the root cause of skin damage caused by ultraviolet rays at the gene level, the present inventors conducted a comprehensive analysis using a DNA microarray. As a result, the expression of annexin, one of the calcium binding protein families, was irradiated with ultraviolet rays. It was found to be significantly reduced in epidermal cells. Almost all cells are said to express at least one protein of the annexin family, but their physiological activity in the skin is unknown. As a property common to the annexin family, it has a phospholipid-binding ability depending on the calcium concentration, and inhibits the activity of phospholipase A ₂ which produces inflammatory mediator leukotriene and prostaglandin precursor arachidonic acid. Annexins 3 and 5 competitively antagonize the protein kinase C substrate, and thus have an activity of inhibiting protein kinase C activity. Furthermore, annexin appears on the cell surface from inside the cell and functions as a receptor protein. For example, Annexin 5 has a high affinity for collagen, and its role in cell adhesion to the basement membrane has attracted attention (Non-Patent Documents 1 to 3).

Davidson F.D. F. et al. Biol. Chem. , 1987, 262, 1698-1705. Schlaepfer D.C. D et al., Biochemistry, 1992, 31, 1886-1891. Kirsch T et al., FEBS Lett. 1992, 310, 143-147.

Annexin is presumed to have similar functions in the skin, since it has a phospholipase A ₂ inhibitory activity, has worked as inhibitors of endogenous in the arachidonic acid cascade, an important to control the inflammation of the skin It is considered a protein. In addition, protein kinase C induces various skin disorders, such as activating proteases that degrade extracellular matrix components, so an annexin that is an endogenous inhibitor may be involved in the protection of ultraviolet skin aging. high. On the other hand, cells cannot function adequately without a scaffold like matrix components, but annexin is a protein necessary for adhesion between cells and matrix components, and acts as an important regulator for maintaining cell functions. It is thought that there is.

  The present inventors have clarified that almost all annexin families are expressed in human epidermal cells, and further found that the expression of these genes is reduced by ultraviolet rays. These results suggest that annexins are useful for the prevention and improvement of skin symptoms thought to be caused by inflammation such as rough skin and conspicuous pores, and application of annexins to anti-aging compositions is expected. The

  However, it is impossible to formulate annexin directly from the viewpoint of stability, safety, and effect, and it is uneconomical. For this reason, a method for activating annexin in the skin or recovering an annexin whose expression is reduced by ultraviolet rays is good, and development of an annexin activator is particularly desired.

Under such circumstances, the present inventors have intensively studied and found that the sodium salt and potassium salt of the compound represented by the chemical formula (1) are stable and show an excellent annexin activating effect. It came to complete.

  The chemical formula (1) of the present invention can be naturally derived, or can be synthesized from tocopherol by a conventional method, and the following documents can also be referred to.

JP-B 61-20583 JP 3-32558

In the present invention, the sodium salt and potassium salt of the compound represented by the chemical formula (1) are used.

  The annexin activator of the present invention can be used for any of pharmaceuticals, quasi drugs, cosmetics or foods. Examples of the dosage form include powders, pills, tablets, injections, suppositories, and emulsions. , Capsules, granules, liquids (including tinctures, fluid extracts, spirits, suspensions, limonades, etc.), lotions, creams, emulsions, gels, aerosols, packs, cleaning agents, bath preparations , Foundation, powder, lipstick, ointment, poultice, paste, plaster, essence, tablet confectionery, beverage and the like. In addition, various components used for normal cosmetics, quasi drugs, pharmaceuticals, etc., for example, oily ingredients, emulsifiers, moisturizers, thickeners, medicinal ingredients, preservatives, pigments, as long as the effects of the present invention are not impaired. , Powders, pH adjusters, ultraviolet absorbers, antioxidants, fragrances and the like can be appropriately blended.

  Examples of the oil component include liquid paraffin, petrolatum, microcrystalline wax, squalane, jojoba oil, beeswax, carnauba wax, lanolin, olive oil, coconut oil, higher alcohol, fatty acid, ester of higher alcohol and fatty acid, and silicone oil. Examples of the emulsifier include nonionic surfactants such as polyoxyethylene alkyl ether, polyoxyethylene fatty acid ester, polyoxyethylene sorbitan fatty acid ester, sorbitan fatty acid ester, glycerin fatty acid ester, polyglycerin fatty acid ester, and polyoxyethylene hydrogenated castor oil. And anionic surfactants such as sodium stearoyl lactate, amphoteric surfactants such as soybean phospholipid, and cationic surfactants such as alkyltrimethylammonium chloride. Examples of the humectant include glycerin, sorbitol, xylitol, maltitol, propylene glycol, polyethylene glycol, 1,3-butylene glycol and the like. Examples of the thickener include clay minerals such as carboxyvinyl polymer, xanthan gum, methyl cellulose, polyvinyl pyrrolidone, gelatin, and bentonite. Examples of medicinal ingredients include various vitamins and derivatives thereof, allantoin, glycyrrhizic acid and derivatives thereof, and various animal and plant extracts.

  The amount of the compound to be blended in the annexin activator of the present invention varies depending on the dosage form and the expected effect, but is usually 0.001% by weight or more, preferably about 0.1 to 50% by weight. Is good. If it is less than 0.001% by weight, a sufficient effect may be difficult to expect, and if it exceeds 50% by weight, an increase in the effect is hardly recognized and it is uneconomical.

  The novel medicinal component of the present invention has an annexin activating effect, controls skin inflammation, and prevents or improves rough skin and conspicuous pores due to keratinization failure. Furthermore, it suppresses the activation of protease and has the effect of maintaining the function of cells, thus preventing skin aging.

  Next, the effects of the present invention will be described with specific examples. These examples specifically illustrate the effect and do not limit the scope of the invention. The compounding quantity in an Example is weight%.

  The medicinal component of the present invention can be formulated as a formulation example as follows.

Formulation Example-1 Beverage Formulation Blending amount (%)
1. dl-α-Tocopheryl sodium phosphate 2.0
2. Citric acid 0.7
3. Fructose dextrose liquid sugar 60.0
4). Perfume appropriate amount 5. Bring the total amount to 100 with purified water.
[Production method] Add 1 to 4 to 5, dissolve by stirring, filter, heat sterilize, and fill into a 50 mL glass bottle.

Formulation Example-2 Beverage Formulation Blending amount (%)
1. d-α-Tocopheryl sodium phosphate 1.0
2. Citric acid 0.7
3. Sucrose 60.0
4). Perfume appropriate amount 5. Bring the total amount to 100 with purified water.
[Production method] Add 1 to 4 to 5, dissolve by stirring, filter, heat sterilize, and fill into a 50 mL glass bottle.

Formulation Example-3 Granule Formulation Blending amount (%)
1. dl-α-Tocopheryl phosphate disodium 3.0
2. Reduced maltose syrup 38.0
3. Microcrystalline cellulose 59.0
[Manufacturing method] An appropriate amount of 70% ethanol is added to components 1 to 3, kneaded, extruded and granulated, and dried to obtain granules.

Formulation Example-4 Tablet Formulation Blending amount (%)
1. d-α-Tocopheryl potassium phosphate 4.0
2. Dried corn starch 25.0
3. Reduced maltose syrup 20.0
4). Powdered sugar 48.0
5. Sucrose fatty acid ester 3.0
[Manufacturing method] An appropriate amount of 70% ethanol is added to components 1 to 4, kneaded, extruded and granulated, and dried to obtain granules. Ingredient 5 is added to the granules and tableted to give 0.52 g tablets.

Formulation example-5 Tablet confection Formulation amount (%)
1. dl-α-Tocopheryl sodium phosphate 0.9
2. Dried corn starch 51.1
3. Erythritol 39.0
4). Citric acid 5.0
5. Sucrose fatty acid ester 3.0
6). Fragrance 1.0
[Manufacturing method] An appropriate amount of water is added to components 1 to 4, kneading, extrusion granulation, and drying to obtain granules. Ingredients 5 and 6 are added to the granules and tableted to obtain 1.0 g of tablet confectionery.

Formulation Example-6 Capsule Formulation Formulation Amount (%)
1. dl-α-tocopheryl sodium phosphate 15.0
2. Cellulose 85.0
[Manufacturing method] Components 1 and 2 are mixed, and a No. 2 hard capsule is filled with 250 mg to obtain a capsule.

Formulation Example-7 Cream Amount (%)
Formula 1. dl-α-Tocopheryl sodium phosphate 2.0
2. Squalane 5.5
3. Olive oil 3.0
4). Stearic acid 2.0
5. Beeswax 2.0
6). Octyldodecyl myristate 3.5
7). Polyoxyethylene cetyl ether (20E.O.) 3.0
8). Behenyl alcohol 1.5
9. Glycerol monostearate2.5
10. Fragrance 0.1
11.1,3-butylene glycol 8.5
12 Ethyl paraoxybenzoate 0.05
13. Methyl paraoxybenzoate 0.2
14 Make the total volume 100 with purified water
[Manufacturing method] Components 2 to 9 are heated and dissolved and mixed, and kept at 70 ° C to obtain an oil phase. Ingredients 1 and 11 to 14 are dissolved by heating and mixed, and kept at 75 ° C. to obtain an aqueous phase. The aqueous phase is added to the oil phase to emulsify, and the mixture is cooled while stirring. The component 10 is added at 45 ° C., and further cooled to 30 ° C. to obtain a product.

Comparative Example-1 Conventional Cream In Formulation Example-7, a product obtained by replacing dl-α-tocopheryl phosphate with purified water was used as a conventional cream.

Formulation example-8 lotion amount (%)
Formula 1. d-α-Tocopheryl sodium phosphate 0.1
2. 1,3-butylene glycol 8.0
3. Glycerin 2.0
4). Xanthan gum 0.02
5. Citric acid 0.01
6). Sodium citrate 0.1
7). Ethanol 5.0
8). Methyl paraoxybenzoate 0.1
9. Polyoxyethylene hydrogenated castor oil (40E.O.) 0.1
10. Perfume proper amount11. Make the total volume 100 with purified water
[Production method] Components 1 to 6 and 11 and components 7 to 10 are uniformly dissolved, mixed and filtered to obtain a product.

Formulation Example-9 Emulsion Formulation Amount (%)
Formula 1. dl-α-Tocopheryl phosphate disodium 0.5
2. Squalane 5.0
3. Olive oil 5.0
4). Jojoba oil 5.0
5. Cetanol 1.5
6). Glycerol monostearate 2.0
7). Polyoxyethylene cetyl ether (20E.O.) 3.0
8). Polyoxyethylene sorbitan monooleate (20E.O.) 2.0
9. Fragrance 0.1
10. Propylene glycol 1.0
11. Glycerin 2.0
12 Methyl paraoxybenzoate 0.2
13. Make the total volume 100 with purified water
[Manufacturing method] Components 2 to 8 are dissolved by heating and mixed, and kept at 70 ° C to obtain an oil phase. Ingredients 1 and 10-13 are dissolved by heating and mixed, and kept at 75 ° C. to form an aqueous phase. The aqueous phase is added to the oil phase to emulsify, and the mixture is cooled while stirring.

Formulation Example -10 Gel formulation (%)
Formula 1. d-α-Tocopheryl potassium phosphate 0.1
2. Ethanol 5.0
3. Methyl paraoxybenzoate 0.1
4). Polyoxyethylene hydrogenated castor oil 0.1
5. Perfume appropriate amount 6.1,3-butylene glycol 5.0
7). Glycerin 5.0
8). Xanthan gum 0.1
9. Carboxyvinyl polymer 0.2
10. Potassium hydroxide 0.2
11. Make the total volume 100 with purified water
[Manufacturing method] Components 2 to 5 and components 1 and 6 to 11 are uniformly dissolved and mixed to obtain a product.

Formulation Example-11 Ointment Amount (%)
Formula 1. dl-α-tocopheryl sodium phosphate 1.0
2. Polyoxyethylene cetyl ether (30E.O.) 2.0
3. Glycerol monostearate 10.0
4). Liquid paraffin 5.0
5. Cetanol 6.0
6). Methyl paraoxybenzoate 0.1
7). Propylene glycol 10.0
8). Make the total volume 100 with purified water
[Manufacturing method] Components 2 to 5 are dissolved by heating and mixed, and kept at 70 ° C to obtain an oil phase. Ingredients 1 and 6 to 8 are dissolved by heating and mixed, and kept at 75 ° C. to form an aqueous phase. The aqueous phase is added to the oil phase to emulsify, and the product is cooled to 30 ° C. with stirring to obtain a product.

Formulation example -12 pack compounding amount (%)
Formula 1. dl-α-Tocopheryl potassium phosphate 0.1
2. Polyvinyl alcohol 12.0
3. Ethanol 5.0
4.1,3-Butylene glycol 8.0
5. Methyl paraoxybenzoate 0.2
6). Paraoxyethylene hydrogenated castor oil (20 EO) 0.5
7). Citric acid 0.1
8). Sodium citrate 0.3
9. Perfume appropriate amount10. Make the total volume 100 with purified water
[Production Method] Components 1 to 10 are uniformly dissolved to obtain a product.

Formulation Example-13 Foundation Formulation (%)
Formula 1. d-α-Tocopheryl phosphate disodium 1.0
2. Stearic acid 2.4
3. Polyoxyethylene sorbitan monostearate (20E.O.) 1.0
4). Polyoxyethylene cetyl ether (20E.O.) 2.0
5. Cetanol 1.0
6). Liquid lanolin 2.0
7). Liquid paraffin 3.0
8). Isopropyl myristate 6.5
9. Butyl paraoxybenzoate 0.1
10. Sodium carboxymethylcellulose 0.1
11. Bentonite 0.5
12 Propylene glycol 4.0
13. Triethanolamine 1.1
14 Methyl paraoxybenzoate 0.2
15. Titanium dioxide 8.0
16. Talc 4.0
17. Bengala 1.0
18. Yellow iron oxide 2.0
19. Perfume proper amount20. Make the total volume 100 with purified water
[Manufacturing method] Components 2 to 9 are heated and dissolved and kept at 80 ° C to obtain an oil phase. Swell component 10 well with component 20, then add components 1 and 11-14 and mix uniformly. To this, components 15 to 18 pulverized and mixed with a pulverizer are added, cooled, component 19 is added at 45 ° C., and cooled to 30 ° C. with stirring to obtain a product.

Formulation Example-14 Bath Agent Formulation (%)
Formula 1. dl-α-tocopheryl sodium phosphate 1.0
2. Sodium bicarbonate 50.0
3. Yellow 202 No. 4 Perfume appropriate amount 5. Make the total amount 100 with anhydrous sodium sulfate
[Production method] Components 1 to 5 are uniformly mixed to obtain a product.

  Next, experimental examples will be given to explain the effects of the present invention in detail.

Experimental Example 1 Expression Analysis of Annexin 5 mRNA A sample was added to normal human keratinocytes in a confluent state so as to be 50 μM, and total RNA was extracted 24 hours later. Annexin 5 mRNA expression level was measured by RT-PCR method based on total RNA. The RT-PCR method is described in TaKaRa RNA PCR Kit (AMV) Ver. 2.1 was used. GAPDH was used as an internal standard. Other operations were carried out according to a predetermined method, and the PCR reaction solution was subjected to agarose gel electrophoresis, and the mRNA expression of annexin 5 and GAPDH was confirmed as a band. These bands were photographed with a polaroid camera and quantified using a densitometer, and the expression level of annexin 5 mRNA was determined as a ratio with respect to the GAPDH mRNA expression level as an internal standard, and expressed as an expression ratio when the control was 1. . As a result, as shown in Table 1, the sodium salt and potassium salt of the compound represented by the chemical formula (1) enhanced the expression of annexin 5 mRNA.

Experimental Example-2 Protective effect of Annexin 5 against ultraviolet irradiation A sample was added to a normal human keratinocyte in a confluent state so as to be 10 μM. After 24 hours, the culture medium was replaced with PBS, and UVB was irradiated with 30 mJ / cm ² . Total RNA was extracted 3 hours after irradiation, and the expression level of annexin 5 mRNA was measured by RT-PCR based on the total RNA. The RT-PCR method is described in TaKaRa RNA PCR Kit (AMV) Ver. 2.1 was used. GAPDH was used as an internal standard. Other operations were carried out according to a predetermined method, and the PCR reaction solution was subjected to agarose gel electrophoresis, and the mRNA expression of annexin 5 and GAPDH was confirmed as a band. These bands were photographed with a polaroid camera and quantified using a densitometer, and the expression level of annexin 5 mRNA was determined as a ratio with respect to the GAPDH mRNA expression level as an internal standard, and expressed as an expression ratio when the control was 1. . As a result, as shown in Table 2, the sodium salt and potassium salt of the compound represented by the chemical formula (1) suppressed the decrease in the expression of annexin 5 mRNA due to ultraviolet rays and exhibited a protective effect.

Experimental Example-3 Use Test Using the cream of Prescription Example-7 and the cream of Comparative Example-1, a use test for 2 months was conducted for 30 women (21-47 years old). After use, a questionnaire survey was conducted on the effect of improving the dryness of the skin, skin irregularities, dullness, and conspicuous pores to determine the cosmetic effect. The evaluation criteria of the questionnaire were evaluated as “excellent” for valid, “good” for slightly effective, “good” for slightly effective, and “impossible” for invalid.

  These results are shown in Table 3. The composition containing the annexin activator of the present invention was excellent in preventing and improving rough skin and conspicuous pores. During the test period, there was no skin problem and there was no problem with safety.

  When similar tests were conducted on Formulation Examples 1 to 6 and 8 to 14, all showed safe and excellent skin aging prevention effects.

The sodium salt and potassium salt of the compound represented by the chemical formula (1) of the present invention have an excellent annexin activating effect on the skin, and dryness, texture disorder, dullness, and conspicuous pores associated with skin keratinization. It can be incorporated into pharmaceuticals, quasi-drugs, cosmetics or foods for the purpose of preventing or improving the symptoms.

Claims (1)

An annexin activator comprising a sodium salt and a potassium salt of a compound represented by the chemical formula (1).