JP2015101539A - External preparation for skin - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an external preparation for skin which increases an enzyme activity associated with the skin condition, and improves the skin condition.SOLUTION: The invention provides an external preparation for skin containing a production acceleration component of heat shock proteins and polyethylene glycol having a weight average molecular weight of 2000-5000. The external preparation for skin of the invention can increase the enzyme activity in a horny layer, and can improve the skin condition. The external preparation for skin of the invention is preferred not to contain an oil solution substantially, and can be used as cosmetics. As a production acceleration component of heat shock proteins, specifically, extract of Curcuma longa, Paeoniae Radix, Salix alba, Glycyrrhiza, and Prunus persica can be illustrated. The root, stem, branch, leaf, seed of the plant having these production acceleration components of heat shock proteins, and a single part of the whole plant may be used, or two or more parts of the whole plant also may be used. Pulverized as it is or dried materials may be used. The extract using a polar solvent as an extracting solvent can be illustrated preferably.

Description

  The present invention relates to an external preparation for skin having an effect of improving the skin condition.

Various enzymes are present in the stratum corneum of the skin, and it has been reported that these enzymes are associated with skin conditions. For example, Patent Document 1 suggests that trypsin-like enzyme activity and chymotrypsin-like enzyme activity in the stratum corneum have a relationship with rough skin.
In addition, caspase-14 (for example, see Non-patent Documents 1 and 2) as an enzyme involved in stratum corneum formation and NMF production, and catalase (for example, a non-patent document) as an enzyme that decreases with aging due to decomposition of hydrogen peroxide. 2 and 3), NADH dehydrogenase (for example, see Non-patent Document 4) as an enzyme involved in energy production, kallikrein-5 (for example, non-related) as an enzyme involved in adhesion of horny layer cells to skin Patent Document 5) is known.

  On the other hand, heat shock proteins are a group of proteins that are expressed when cells are exposed to stress such as heat and ultraviolet rays and protect cells, and are named according to their molecular weights, for example, HSP60, 70 and 90 refer to proteins having molecular weights of 60, 70, and 90 kDa, respectively. With regard to heat shock protein, skin external preparations containing yeast extract that produces heat shock protein and heat shock protein factor protect the skin from external stimuli such as ultraviolet rays and heat, repair damaged skin, Improvement has been reported (see, for example, Patent Document 2). However, there are cases where the skin condition does not improve with a topical skin preparation containing only ingredients that produce heat shock protein factor using these techniques, and there is a need for a technique for repairing and improving damaged skin. It was.

JP-A-8-68791 JP 2004-331602 A

NatureCell Biology 2007 (6) p666-674 Collection of experimental protocols for developing functional cosmetic materials (CMC Publishing) Experimental Gerontology 2007 (42) p924-929 J Cosmet Dermatol. 2012 (11) No. 1 p3-8 Biol. Chem. 2008 (389) No. 6p669-680

  This invention is made | formed in such a condition, and makes it a subject to provide the skin external preparation which improves the enzyme activity relevant to a skin state, and improves a skin state.

The present inventors have advanced research to provide an external preparation for skin that can improve the skin condition, and surprisingly, skin containing a heat shock protein production promoting component and polyethylene glycol having a weight average molecular weight of 2000 to 5000. It was found that an external preparation suppresses and repairs enzyme denaturation due to ultraviolet rays and the like, reduces a decrease in enzyme activity, and improves the skin condition, thereby completing the present invention. That is, the present invention is as follows.
<1> A heat shock protein production-promoting ingredient and a polyethylene glycol having a weight average molecular weight of 2000 to 5000, <2> a skin external preparation <2> substantially free of an oil agent <1 > The skin external preparation described in <3>, wherein the skin condition improved by the skin external preparation is at least one selected from the group consisting of skin brightness, pore conspicuity, wrinkles, and skin irregularities <1> or <2> skin external preparation <4> cosmetic preparation, <1> to <3> skin external preparation

It is a graph which shows the relationship between kallikrein-5 activity and skin brightness. It is a graph which shows the relationship between kallikrein-5 activity and a pore score. It is a graph which shows the relationship between kallikrein-5 activity and the number of wrinkles. It is a graph which shows the relationship between kallikrein-5 activity and the unevenness | corrugation of skin.

  Hereinafter, the skin external preparation of the present invention will be described.

(1) Heat shock protein production promoting component as an essential component of the external preparation for skin of the present invention The external preparation for skin of the present invention contains a heat shock protein production promoting component as its essential component. The heat shock protein production promoting component is not particularly limited as long as it can be used as a heat shock protein production promoting component in a topical skin preparation. Specifically, extraction of turmeric, glaze, white willow, licorice, peach, etc. Plant extracts such as products can be exemplified, and a single part of the root, stem, branch, leaf, seed, whole plant of the plant used as these extracts may be used, or two or more parts may be used. You may use what was grind | pulverized or dried as it is. An extract using a polar solvent as the extraction solvent can be preferably exemplified. Examples of the extraction solvent include water, ethanol, isopropyl alcohol, butanol, and other polyhydric alcohols such as 1,3-butanediol and polypropylene glycol, acetone, Extracts extracted with one or two or more polar solvents selected from ketones such as methyl ethyl ketone, ethers such as diethyl ether and tetrahydrofuran, and those obtained by drying the extract into powder it can. These pulverized product, extract, dried product and the like can be purified by partitioning between columns and solutions to increase the active ingredients.

As a turmeric, an extract of the rhizome of Curcuma domestica Val. As licorice, Glycyrrhiza root extract of leguminous licorice genus, Prunus Persica of rosaceae peach genus as peach
Batsch leaf extract is particularly preferred.

  The heat shock protein production promoting component is 0.0001% by mass to 10% by mass, more preferably 0.001% by mass to 5% by mass, and still more preferably 0.015% by mass with respect to the total amount of the external preparation for skin of the present invention. % To 3% by mass is preferable. If this is less than the lower limit, the skin condition improvement effect of the external preparation for skin of the present invention is not exhibited, and if the upper limit is exceeded, the effect reaches its peak, causing problems such as color and odor, and when used as an external preparation for skin This is because the degree of freedom may be lost.

(2) Polyethylene glycol which is an essential component of the external preparation for skin of the present invention The external preparation for skin of the present invention contains polyethylene glycol having a weight average molecular weight of 2000 to 5000 as an essential component. Specific examples of such polyethylene glycol include polyethylene glycol 2000 and polyethylene glycol 4000. Since these are commercially available products, they can be obtained and used. Specific examples of commercially available products include “Toho Polyethylene Glycol 2000” and “PEG 4000” (both manufactured by Toho Chemical Co., Ltd.).

  In the external preparation for skin of the present invention, the polyethylene glycol having a weight average molecular weight of 2000 to 5000 is preferably contained in an amount of 0.012 to 5.0% by mass based on the total amount of the external preparation for skin, 0.51 to 3.0. It is more preferable to contain by mass. If the content is less than the lower limit, the effect of improving the enzyme activity in the stratum corneum may not be observed. If the content exceeds the upper limit, the effect reaches a peak, and the feel as a skin external preparation may be deteriorated.

  More preferably, the external preparation for skin of the present invention is characterized in that it usually contains substantially no oil used for the external preparation for skin. However, in the present invention, the external preparation contains no oil. Even if it does, it means that the content is 0.5 mass% or less with respect to the skin external preparation whole quantity.

Further, the oil agent in the present invention specifically includes macadamia nut oil, avocado oil, corn oil, olive oil, rapeseed oil, sesame oil, castor oil, safflower oil, cottonseed oil, jojoba oil, coconut oil, palm oil, liquid lanolin. Oils such as hardened coconut oil, hardened oil, mole, hardened castor oil, beeswax, candelilla wax, carnauba wax, ibotarou, lanolin, reduced lanolin, hard lanolin, jojoba wax, waxes, liquid paraffin, squalane, pristane, ozokerite, paraffin , Hydrocarbons such as ceresin, petrolatum and microcrystalline wax, higher alcohols such as cetyl alcohol, stearyl alcohol, isostearyl alcohol, behenyl alcohol, octyldodecanol, myristyl alcohol, cetostearyl alcohol Higher fatty acids such as lauric acid, myristic acid, stearic acid, behenic acid, cetyl isooctanoate, isopropyl myristate, hexyldecyl isostearate, diisopropyl adipate, di-2-ethylhexyl sebacate, cetyl lactate, diisostearyl malate , Ethylene glycol di-2-ethylhexanoate, neopentyl glycol dicaprate, glycerin di-2-heptylundecanoate, glycerin tri-2-ethylhexanoate, trimethylolpropane tri-2-ethylhexanoate, triisostearic acid tri Synthetic ester oils such as methylolpropane, penta-2-ethylhexanoic acid pentane erythritol, fatty acid soap (sodium laurate, sodium palmitate, etc.), potassium lauryl sulfate, alkyl Anionic surfactants such as acid triethanolamine ether, cationic surfactants such as stearyltrimethylammonium chloride, benzalkonium chloride, laurylamine oxide, imidazoline-based amphoteric surfactants (2-cocoyl-2-imidazolinium) Hydroxide-1-carboxyethyloxy disodium salt, etc.), betaine surfactants (alkyl betaines, amide betaines, sulfobetaines, etc.), amphoteric surfactants such as acylmethyl taurine, sorbitan fatty acid esters (sorbitan monostears) Rate, sorbitan sesquioleate, etc.), glycerin fatty acids (such as glyceryl monostearate), propylene glycol fatty acid esters (such as propylene glycol monostearate), hardened castor oil derivatives, glycerin alkyl Ether, POE sorbitan fatty acid esters (POE sorbitan monooleate, polyoxyethylene sorbitan monostearate, etc.), POE sorbite fatty acid esters (POE-sorbite monolaurate, etc.), POE glycerin fatty acid esters (POE-glycerin monoiso) Stearates, etc.), POE fatty acid esters (polyethylene glycol monooleate, POE distearate, etc.), POE alkyl ethers (POE2-octyldodecyl ether, etc.), POE alkyl phenyl ethers (POE nonylphenyl ether, etc.), pluronic types, POE / POP alkyl ethers (POE / POP2-decyltetradecyl ether, etc.), Tetronics, POE castor oil / hardened castor oil derivatives (POE castor oil, P E-cured castor oil, etc.), nonionic surfactants such as sucrose fatty acid ester, alkyl glucoside, dimethicone, phenyl dimethicone, polyether-modified silicone, polyglycerin-modified silicone, amino-modified silicone and other silicones, diethylaminohydroxybenzoylbenzoate Examples thereof include UV absorbers such as hexyl acid, t-butylmethoxybenzoylmethane, 2-ethylhexyl paramethoxycinnamate, dimethicodiethylbenzalmalonate, and homomenthyl salicylate, and oil-soluble vitamins.

Furthermore, the external preparation for skin of the present invention can contain optional components other than oils, which are usually used in external preparations for skin, as long as the effects are not impaired. Examples of such optional components include polyethylene glycol having a weight average molecular weight of less than 2000, polyhydric alcohols of polyethylene glycol having a weight average molecular weight of more than 5000, moisturizing components such as sodium pyrrolidone carboxylate, lactic acid, sodium lactate, guar gum, quince seed Carrageenan, galactan, gum arabic, pectin, mannan, starch, xanthan gum, curdlan, methylcellulose, hydroxyethylcellulose, carboxymethylcellulose, methylhydroxypropylcellulose, chondroitin sulfate, dermatan sulfate, glycogen, heparan sulfate, hyaluronic acid, sodium hyaluronate, Tragacanth gum, keratan sulfate, chondroitin, mucoitin sulfate, hydroxyethyl guar gum, carboxymethyl gum Increase in gum, dextran, keratosulfuric acid, locust bean gum, succinoglucan, caronic acid, chitin, chitosan, carboxymethylchitin, agar, polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymer, sodium polyacrylate, polyethylene glycol, bentonite, etc. Viscosity agents, polyhydric alcohols other than alkanediols of the present invention such as 1,3 butanediol, glycerin, diglycerin, etc., which may be surface treated, mica, talc, kaolin, synthetic mica, calcium carbonate, magnesium carbonate, anhydrous Powders such as silicic acid (silica), aluminum oxide, barium sulfate, surface treatment, cobalt oxide, ultramarine blue, bitumen, zinc oxide inorganic pigments, surface treatment, iron oxide dioxide Composite such as titanium sintered body A surface treatment, fine particle titanium dioxide having an average primary particle size of 100 μm or less, fine particle zinc oxide, a pearl agent such as titanium mica, fish phosphorus foil, bismuth oxychloride, which may be surface treated, Red 202, Red 228, Red 226, Yellow 4, Blue 404, Yellow 5, Red 505, Red 230, Red 223, Orange 201, Red 213 which may be raked Organic powders such as yellow 204, yellow 203, blue 1, green 201, purple 201, red 204, polyethylene powder, polymethyl methacrylate, nylon powder, organopolysiloxane elastomer, etc. , Lower alcohols such as ethanol and isopropanol, water-soluble vitamins such as ascorbic acid and ascorbic acid derivatives, and the like.

(3) The skin external preparation of this invention The skin external preparation of this invention can improve the enzyme activity in a stratum corneum, and can improve a skin state. Examples of the enzyme in the stratum corneum include kallikrein-5, caspase-14, catalase, and NADH dehydrogenase, histidine ammonia lyase, and transglutaminase, and more preferably kallikrein-5, caspase-14, catalase, and NADH dehydrogenase. .
If at least one enzyme activity selected from the group consisting of these enzymes is improved, it can be determined that the skin condition has improved.
The enzyme activity is measured, for example, by extracting the enzyme from the stratum corneum cells collected with a commercially available adhesive tape or the like with a buffer, and measuring the amount of change in enzyme activity of the obtained enzyme solution according to a conventional method. It is done. The stratum corneum collection site is not particularly limited.
Whether or not the enzyme activity is improved can be determined by measuring how the enzyme activity changes before and after the use of the external preparation for skin of the present invention.
Hereinafter, the present invention will be described in detail with reference to test examples.

Test example

(Extraction of enzyme in stratum corneum)
For one healthy person, the face was double-washed (make-up / face-wash), and then the stratum corneum cells of the face were collected by the tape stripping method.
The collected stratum corneum was homogenized with an enzyme extract (0.1 M Tris-HCl (pH 7.5) +0.14 M NaCl + 0.1% Tween 20), and the supernatant obtained by centrifugation at 15000 rpm was used as the enzyme solution.
The protein concentration required for calculation of enzyme activity was measured using Protein Assay Kit (Dojin Chemical) for the enzyme solution.
In this test, kallikrein-5, caspase-14, catalase, and NADH dehydrogenase were selected as the enzymes in the stratum corneum, and the enzyme activity of each enzyme was measured.

(Measurement of enzyme activity in stratum corneum)
Kallikrein-5 was measured by the following method. That is, MCA substrate (Boc-Val-Pro-Arg-MCA, Dojindo) (0.2 mM) 20 μL, assay buffer (50 mM HEPES (pH 7.5) +60 mM)
NaCl + 0.01% Chaps + 5 mM EDTA + 2 mM DTT + 1.5 mM sodium citrate) 130 μL, enzyme extract (without enzyme) 30 μL is incubated in a small tube for 10 min at 37 ° C. to blank and similarly MCA substrate (0.2 mM ) Add 20 μL of assay buffer (50 mM HEPES (pH 7.5) +60 mM NaCl + 0.01% Chaps + 5 mM EDTA + 2 mM DTT + 1.5 mM sodium citrate) 130 μL and incubate for 10 minutes at 37 ° C. in a small tube. did.
For both the blank and the sample, the reaction was stopped with a 0.1 M sodium monochloroacetate solution (pH 4.3), the fluorescence intensity (EX 370 nm, EM 460 nm) was measured, and the enzyme activity was calculated.

Caspase-14 was measured by the following method. Specifically, 20 μL of MCA substrate (0.2 mM) and 160 μL of assay buffer (50 mM HEPES (pH 7.5) +60 mM NaCl + 0.01% Chaps + 5 mM EDTA + 2 mM DTT + 1.5 mM sodium citrate) were incubated in a small test tube for 10 minutes at 37 ° C. and blanked. Similarly, 30 μL of enzyme solution was added to 160 μL of MCA substrate (0.2 mM) 20 μL, assay buffer (50 mM HEPES (pH 7.5) +60 mM NaCl + 0.01% Chaps + 5 mM EDTA + 2 mM DTT + 1.5 mM sodium citrate), and a small test tube. Incubated for 10 minutes at 37 ° C. to give a sample.
For both the blank and the sample, the reaction was stopped with a 0.1 M sodium monochloroacetate solution (pH 4.3), measured with fluorescence intensity (EX 370 nm, EM 460 nm), and the enzyme activity was calculated.

Catalase was measured by the following method. That is, 300 μL of 0.1 M Tris-HCl (pH 7.5) +0.14 M NaCl + 0.1% Tween 20 was mixed with 300 μL of 50 mM potassium phosphate buffer (pH 7.2) in a test tube to obtain a blank. Similarly, 300 μL of enzyme solution was mixed with 300 μL of potassium phosphate buffer (pH 7.2) to prepare a sample. After adding 300 μL of 31 mM hydrogen peroxide solution to each sample solution prepared in a test tube and reacting at 25 ° C., the decrease in absorbance at 240 nm derived from hydrogen peroxide for 0 to 4 minutes was traced. The amount of decrease in absorbance per minute was calculated. The enzyme activity (μmol / min / g) was determined from the molecular absorbance coefficient of hydrogen peroxide at 240 nm (1 μmol / mL = 0.036).

NADH dehydrogenase was measured by the following method. Specifically, 2.8 mL of 20 mM Tris-HCl buffer (pH 7.5) containing 0.2 mM of NADH as a substrate was preheated at 25 ° C. for 5 minutes, and then 1.2 mM 2,6-dichlorophenolindophenol (DCIP). 0.1 mL of an aqueous solution was added, and then 0.1 mL of an enzyme solution previously diluted with an enzyme diluent (200 mM Tris-HCl buffer (pH 7.5)) was added to start the reaction, and every 20 seconds at 25 ° C. The decrease in absorbance at 600 nm was measured for 4 minutes.
As a blind test, in the above, 0.1 mL of enzyme dilution solution was added instead of 0.1 mL of NADH dehydrogenase solution, and the reaction was started by operating in the same manner as described above. Absorbance measured in minutes was measured.

<Test Example 1>
(Measurement of enzyme activity of samples 1 to 12)
After irradiating ultraviolet rays (UVB) to 100 mJ / cm <2> in order to damage the enzyme, heat shock protein production promoting components and polyethylene glycol having a weight average molecular weight of 2000 to 5000 are added to the enzyme solution in Table 1. It added so that it might become a density | concentration (w / v), and the enzyme activity of kallikrein-5, a caspase-14, a catalase, and a NADH dehydrogenase after 3 hours was measured. The enzyme activity of each sample when the control was 100% was calculated and displayed in%. Moreover, the activity of each enzyme when not irradiating with ultraviolet rays was 170 to 180% with respect to 100% of the control.

From the results of Table 2, in the group of samples 1 to 4 containing heat shock protein production promoting component and polyethylene glycol having a weight average molecular weight of 2000 to 5000, the activity of the enzyme due to UV damage compared to the control and samples 5 to 12 The decrease was suppressed. Therefore, the skin external preparation of the present invention containing a heat shock protein production promoting component and a polyethylene glycol having a weight average molecular weight of 2000 to 5000 suppresses and repairs enzyme denaturation due to ultraviolet rays, and suppresses a decrease in enzyme activity. It became clear that it improved.

<Test Example 2>
(Relationship between enzyme and skin condition)
In order to grasp the relationship between the enzyme of kallikrein-5 and the skin condition, as described above, the stratum corneum cells of the face of 60 healthy subjects were collected and the enzyme activity was measured. A medium-high group was set, and it was observed whether there was a difference in skin brightness, pore conspicuousness, wrinkles, and unevenness (unevenness) in the low and high groups.
After cleansing the face (make-up / face wash), rest at room temperature of 22 ± 2 ° C and humidity of 50 ± 5%. The right cheek lower part is the spectrocolorimeter CM-2600d (Konica Minolta Optics). Co., Ltd.) measured L * value, measured 5 times, and determined the average value.
For conspicuous pores, wrinkles, and skin irregularities, wash the face in the same manner as described above, and after resting under the conditions of the previous period, VISIA for the front face and three diagonal parts
The number of pore scores, irregularities and wrinkles was measured using Evolution (Canfield Scientific Ltd.).

1-4 show the results of skin brightness (L * value), pore conspicuousness (pore score), number of wrinkles, and skin irregularities (unevenness) in the groups with low and high enzyme activity. The group with high enzyme activity was observed to have lighter skin, lower pore score, less conspicuous pores, fewer wrinkles, and fewer irregularities than the lower group. Therefore, it has been found that an increase in enzyme activity leads to an improvement in skin condition. Similarly, in caspase-14, catalase, and NADH dehydrogenase, it was confirmed that an increase in enzyme activity leads to an improvement in skin condition.

Hereinafter, the present invention will be described in detail with reference to examples, but it goes without saying that the present invention is not limited to such examples.

According to the formulation shown in Table 3, the external preparation for skin of the present invention was prepared.
Ingredient (a) in Table 3 was heated to 60 ° C. with stirring. To this was added the component (b) stirred and mixed at room temperature, and then (c) was added and stirred continuously to obtain a homogeneous solution. Next, the ingredient (e) was added and stirred and mixed, and stirring was continued until it became uniform to obtain a skin external preparation. In addition, the number in component (I)-(E) in a table | surface represents the mass%.

  ADVANTAGE OF THE INVENTION According to this invention, the skin external preparation which improves a skin state can be provided.

Claims (4)

An external preparation for skin, comprising a heat shock protein production promoting component and polyethylene glycol having a weight average molecular weight of 2000 to 5000 The skin external preparation according to claim 1, which contains substantially no oil. The skin condition improved by the external preparation for skin is at least one selected from the group consisting of skin brightness, conspicuous pores, wrinkles, and skin irregularities. Topical skin preparation The external preparation for skin according to claim 1, which is a cosmetic.

Images