JP7497939B2 - Anti-dull skin - Google Patents

Description

The present invention relates to an agent for improving dullness.

Skin dullness is caused by a combination of internal factors such as aging and external factors such as ultraviolet rays and low humidity. In particular, it is known that skin dullness caused by ultraviolet radiation and aging is mainly caused by an increase in melanin in the epidermis and glycation of dermal collagen.
Oral preparations or topical skin preparations that suppress the accumulation of such melanin and advanced glycation end products (AGEs) and improve dull skin have been disclosed (Patent Documents 1 to 5).

The inventors also discovered for the first time that a drop in air pressure is involved in dull skin (Non-Patent Document 1).

JP 2011-102270 A JP 2011-195495 A JP 2014-114233 A JP 2016-210729 A JP 2018-123090 A

POLA ORBIS HOLDINGS Co., Ltd. News Release dated November 12, 2018

Dullness caused by a drop in atmospheric pressure is a temporary dullness of the skin caused by poor circulation due to a drop in oxygen concentration in the blood and disturbances in the autonomic nervous system, and is therefore different from dullness caused by melanin and AGEs that accumulate over a long period of time.

As described above, since there are many factors that cause dullness, it has been difficult to obtain a truly satisfactory anti-dullness effect simply by blending an ingredient that can prevent or improve one of the above-mentioned factors that cause dullness.
In view of the above circumstances, an object of the present invention is to provide a dullness improving agent that acts in a multifaceted manner on temporary dullness caused by a drop in atmospheric pressure and on dullness caused by ultraviolet rays and aging.

The present invention, which solves the above problems, provides an agent for improving dullness, which contains an inhibitor of an increase in calcium ion concentration in vascular smooth muscle cells due to endothelin stimulation, and which contains at least one component selected from a melanin production inhibitor and an AGEs production inhibitor.
The present invention also relates to an agent for improving dullness, which comprises an ingredient that inhibits an increase in calcium ion concentration in vascular smooth muscle cells due to endothelin stimulation, an ingredient that inhibits melanin production, and an ingredient that inhibits AGEs production.
The skin dullness improving agent of the present invention contains two or three skin dullness improving components, and therefore has a superior anti-skin dullness effect as compared with a skin dullness improving agent containing only one skin dullness improving component.

In a preferred embodiment of the present invention, the component that inhibits the increase in calcium ion concentration in vascular smooth muscle cells due to endothelin stimulation is an extract of a plant of the genus Chamaemelum (Chamaemelum L.) of the Asteraceae family.

In a preferred embodiment of the present invention, the melanin production inhibitor is an ingredient selected from ascorbic acid and its derivatives.

In a preferred embodiment of the present invention, the AGEs production inhibitor is a fermented product obtained by fermenting the calyx of a plant of the genus Hibiscus (Malvaceae) with lactic acid bacteria.

The present invention provides a method for screening for components that improve dull skin caused by a drop in atmospheric pressure, using as an index the inhibitory effect on an increase in calcium ion concentration in vascular smooth muscle cells due to endothelin stimulation.
The present invention also relates to an agent for improving dullness caused by a drop in atmospheric pressure, which comprises a component screened by the above screening method.

The present invention is a skin dullness improving agent that contains a skin dullness improving component screened by the above screening method and contains at least one component selected from a melanin production inhibitor and an AGEs production inhibitor.

The skin dullness improving agent of the present invention acts on the causes of skin dullness from multiple angles, and is excellent at comprehensively preventing or improving skin dullness.

FIG. 1 is a diagram showing the average brightness calculated from photographed images of vascular smooth muscle cells when Roman chamomile extract was added in Test Example 1.

<1> Regarding the component that inhibits an increase in calcium ion concentration in vascular smooth muscle cells due to endothelin stimulation The component of the present invention that inhibits an increase in calcium ion concentration in vascular smooth muscle cells due to endothelin stimulation can be determined by the screening method described below. It is preferably an extract of a plant of the genus Chamaemelum L. in the family Asteraceae, and particularly preferably an extract of Roman chamomile (Chamaemelum nobile).

There are no particular limitations on the part of the plant of the genus Camaemerum in the Asteraceae family that may be extracted, and one or more parts selected from the whole plant, leaves, stems, flowers, fruits, and roots may be used, with the flower being particularly preferred.

When extracting, it is preferable to process the plant of the genus Camaemerum of the Asteraceae family or its dried matter in advance by crushing or cutting it into small pieces to improve the extraction efficiency. Examples of extraction methods include a method in which the plant is immersed in an extraction solvent at room temperature, cooled or heated under normal pressure, or under pressure or reduced pressure, a method in which the plant is distilled using a method such as steam distillation, and a squeezing method in which the plant of the genus Camaemerum of the Asteraceae family is squeezed to obtain an extract. These methods can be used alone or in combination of two or more types to perform the extraction.

The extraction solvent is preferably a polar solvent, and suitable examples include one or more selected from water, alcohols such as ethanol, isopropyl alcohol, and butanol, polyhydric alcohols such as 1,3-butanediol and polypropylene glycol, ketones such as acetone and methyl ethyl ketone, and ethers such as diethyl ether and tetrahydrofuran, and among these, it is preferable to use 1,3-butanediol.

When extracting by immersion, after immersion, insoluble matters can be removed by filtration, and then the solvent can be removed by vacuum concentration or the like, but removal by freeze-drying is particularly preferred. The powder composition from which the solvent has been removed may be used as is in the powder state, or it may be fractionated and purified by liquid-liquid extraction with water and ethyl acetate, water and butanol, or the like, or by column chromatography filled with silica gel or ion exchange resin.

The content of the component that inhibits the increase in calcium ion concentration in vascular smooth muscle cells due to endothelin stimulation is preferably 0.00001 to 0.15% by mass, more preferably 0.0001 to 0.1% by mass, and even more preferably 0.0005 to 0.05% by mass (dry mass in the case of an extract) based on the total amount of the skin dullness improving agent.

<2> Ingredients that inhibit melanin production Activation of pigment cells (melanocytes) in the skin leads to a marked increase in melanin production, resulting in age spots, freckles, and pigmentation after sunburn. Accumulation of melanin in the skin leads to increased dullness of the skin and a decrease in skin transparency.

The melanin production-inhibiting component of the present invention is preferably a component that acts on the melanin synthesis pathway and inhibits melanin production, thereby preventing and improving dullness caused by the accumulation of melanin.
Examples of melanin production inhibitors used in the present invention include tyrosinase activity inhibitors such as arbutin, kojic acid, ellagic acid, rucinol, mulberry extract, angelica extract, sanguisorba officinalis extract, and oil-soluble licorice extract, as well as ascorbic acid and its derivatives, with ascorbic acid derivatives being particularly preferred.

Ascorbic acid derivatives are not particularly limited as long as they are generally known to have ascorbic acid as a basic skeleton, and examples thereof include ascorbic acid glucoside, ascorbic acid phosphate ester, ascorbic acid sulfate ester, ascorbic acid fatty acid ester, and ascorbic acid alkyl ether, and it is particularly preferable to use ascorbic acid glucoside for the inhibition of melanin production.

The content of the melanin production inhibitor is preferably 0.00001 to 10% by mass, more preferably 0.0001 to 8% by mass, and even more preferably 0.0005 to 6% by mass (dry mass in the case of an extract) based on the total amount of the dullness improving agent.

<3> About ingredients that inhibit AGEs production The final products obtained by glycation of proteins are called advanced glycation end products (AGEs). AGEs form crosslinks between themselves or between AGEs and biological proteins, causing them to aggregate and become difficult to break down. When glycation occurs in the dermis of the skin, collagen fibers harden, reducing the firmness and elasticity of the skin, inducing dullness due to yellowing, and reducing the permeability of the skin.

The AGE production inhibiting component of the present invention is preferably a component that has the effect of preventing and improving dull skin caused by AGEs by inhibiting the production or accumulation of AGEs or promoting the decomposition of AGEs.
The AGEs production inhibitor used in the present invention is not particularly limited as long as it is an ingredient having an anti-AGEs effect, and examples thereof include the fruit of wild grape, the style of corn, olive leaf extract, tea extract, water chestnut extract, Budreja axillaris leaf extract, henna flower extract, a fermented product obtained by fermenting the calyx of a plant of the genus Hibiscus (Hibiscus L.) of the family Malvaceae with lactic acid bacteria (hereinafter simply referred to as a hibiscus fermented product), AGEs production inhibitors such as 4-vinylcatechol, AGEs cross-linked structure cleavage ingredients such as peony root extract, AGEs cross-linking inhibitors such as magnesium aspartate, zinc gluconate, copper gluconate, and among these, it is preferable to use a hibiscus fermented product.

The plants of the genus Hibiscus in the family Malvaceae that can be used in the present invention include, for example, roselle (Hibiscus sabdariffa L.), rose of Sharon (Hibiscus syriacus), Hibiscus mutabilis, Hibiscus coccineus, Hibiscus tiliaceus, and Hibiscus schizopetalus, with roselle (Hibiscus sabdariffa L.) being preferred.

Lactic acid bacteria used for fermentation include, for example, lactic acid bacteria of the genus Lactobacillus, such as Lactobacillus plantarum, Lactobacillus brevis, and Lactobacillus casei; lactic acid bacteria of the genus Carnobacterium, such as Carnobacterium divergens and Carnobacterium pisicola; lactic acid bacteria of the genus Carnobacterium, such as Leuconostoc mesenteroides and Leuconostoc citreum; lactic acid bacteria of the genus Leuconostoc, such as Streptococcus faecalis and Streptococcus pyogenes; lactic acid bacteria of the genus Enterococcus, such as Enterococcus caseiflavus and Enterococcus sulfureus; lactic acid bacteria of the genus Enterococcus, such as Lactococcus plantarum and Lactococcus raffinolactis; Lactococcus genus lactic acid bacteria such as Weissella confusa and Weissella kandleri; Atopobium minutum and Atopobium parvulus lactic acid bacteria; Vagococcus genus lactic acid bacteria such as Vagococcus fluvialis and Vagococcus salmoninarum; Pediococcus Examples of lactic acid bacteria include lactic acid bacteria of the genus Pediococcus, such as Pediococcus damnosus and Pediococcus pentosaceus. Among these lactic acid bacteria, it is preferable to use Lactobacillus plantarum, since it is not extremely anaerobic and is easy to handle.

Hibiscus fermentation products can be produced by the method described in Japanese Patent No. 5,129,440, in which calyxes of a plant of the genus Hibiscus, dried and pulverized as necessary, are immersed or suspended in a fermentation medium, and then lactic acid bacteria are inoculated into the suspension and fermented.
In the present invention, a commercially available extract can be used as the extract of the fermented hibiscus product, and a suitable example of the extract is fermented hibiscus flower extract.

The content of the AGEs production inhibitor is preferably 0.00001 to 10% by mass, more preferably 0.0001 to 8% by mass, and even more preferably 0.0005 to 6% by mass (dry mass in the case of an extract) based on the total amount of the dullness improving agent.

<4> Regarding the skin dullness improving agent of the present invention The skin dullness improving agent of the present invention is an agent for improving skin dullness that contains an inhibitor of an increase in calcium ion concentration in vascular smooth muscle cells due to endothelin stimulation, and also contains a melanin production inhibitor and/or an AGEs production inhibitor.
In the present invention, it is preferable to include an extract of a plant of the genus Camaemellum of the Asteraceae family as an inhibitor of the increase in calcium ion concentration in vascular smooth muscle cells due to endothelin stimulation, an ascorbic acid derivative as a melanin production inhibitor, and a hibiscus fermentation product as an AGEs production inhibitor.

In the skin dullness improving agent of the present invention, the content ratio of the extract of a plant of the genus Camaemerum of the Asteraceae family, the ascorbic acid derivative, and the hibiscus fermentation product is preferably 0.001 to 20 parts by mass of the ascorbic acid derivative and 0.00005 to 0.01 parts by mass of the hibiscus fermentation product, and more preferably 0.01 to 10 parts by mass of the ascorbic acid derivative and 0.0001 to 0.008 parts by mass of the hibiscus fermentation product, relative to 0.001 part by mass of the extract of a plant of the genus Camaemerum of the Asteraceae family (dry mass in the case of an extract).
In addition, a suitable example of the content ratio of the extract of a plant of the genus Camaemerum of the Asteraceae family, the ascorbic acid derivative, and the hibiscus fermentation product in the present invention is 0.0005-0.1:0.5-5:0.0005-0.1 (in the case of an extract, dry mass).

The skin dullness improving agent of the present invention can be produced by mixing each component with a base and other components that are typically used in skin topical preparations, medical products, and food and beverage products.

The dullness improving agent of the present invention is preferably in the form of an external preparation or an oral preparation.External preparations include, for example, cosmetics, quasi-drugs, skin external medicines, etc.Also, oral preparations include supplements with dosage forms such as tablets, granules, and drinks.Among them, the form of cosmetics that can be used continuously is preferred, and particularly the form of lotion, beauty essence, milky lotion, cream, gel, sun care product, etc. is preferred.

Skin topical compositions may contain optional ingredients that are commonly used. Suitable examples of such optional components include hydrocarbons such as squalane, petrolatum, and microcrystalline wax; esters such as jojoba oil, carnauba wax, and octyldodecyl oleate; triglycerides such as olive oil, beef tallow, and coconut oil; fatty acids such as stearic acid, oleic acid, and retinoic acid; higher alcohols such as oleyl alcohol, stearyl alcohol, and octyldodecanol; anionic surfactants such as sulfosuccinate esters and sodium polyoxyethylene alkyl sulfates; amphoteric surfactants such as alkyl betaine salts; cationic surfactants such as dialkyl ammonium salts; sorbitan fatty acid esters, fatty acid monoglycerides, polyoxyethylene adducts thereof, nonionic surfactants such as polyoxyethylene alkyl ethers and polyoxyethylene fatty acid esters; polyhydric alcohols such as polyethylene glycol, glycerin, and 1,3-butanediol; thickening and gelling agents; antioxidants; ultraviolet protection agents such as ultraviolet absorbing agents and ultraviolet scattering agents; coloring agents; preservatives; pH adjusters; powders; and various active ingredients.

The skin dullness improving agent of the present invention can be used to prevent or improve temporary skin dullness caused by a drop in atmospheric pressure, as well as skin dullness caused by ultraviolet rays, aging, and the like.
For example, it is preferable to apply the present invention to individual or multiple factors such as pale complexion due to vasoconstriction, pigmentation due to accumulation of melanin, loss of skin transparency, and yellowing of the skin due to accumulation of AGEs.

<5> Screening method using as an index the inhibitory effect on an increase in calcium ion concentration in vascular smooth muscle cells The present invention also provides a screening method for components that improve dullness caused by a decrease in atmospheric pressure, using as an index the inhibitory effect on an increase in calcium ion concentration in vascular smooth muscle cells due to endothelin stimulation.
In the present invention, among the causes of skin dullness, attention is focused on temporary skin dullness caused by vasoconstriction due to a drop in atmospheric pressure. Vasoconstriction occurs when endothelium-derived contracting factor (EDCF) is released from vascular smooth muscle cells, causing an increase in intracellular calcium ion concentration.

The screening method of the present invention uses as an index the amount of calcium ions that increases upon addition of endothelin-1 to cultured cells.
In the screening method of the present invention, a component to be screened (hereinafter referred to as a test substance) is added to a culture medium containing cultured cells in vitro, and then a calcium indicator and endothelin-1, a calcium ion inducer, are added to the cultured cells.

Next, the amount of calcium ions in the cells is measured using the fluorescence of the calcium indicator as an indicator, and if the measured amount of calcium ions is smaller than the amount of calcium ions in cells to which the test substance was not added, it is determined that the test substance has an effect on the increase in intracellular calcium ion concentration due to endothelin stimulation.

The cultured cells used in the screening of the present invention are preferably cultured cells derived from vascular smooth muscle, and may be, for example, a cell line established from vascular smooth muscle cells collected from a living body, or a cell line induced to differentiate into vascular smooth muscle cells from pluripotent stem cells (e.g., iPS cells, ES cells).

The amount of calcium ions in the cytoplasm can be measured using a calcium indicator by a standard method.
The calcium indicator may be any reagent capable of detecting a signal correlated with the amount of calcium ions present, and may be, for example, a fluorescent indicator, a luminescent chemiluminescent indicator, or an absorbance indicator that reacts quantitatively with calcium ions. Commercially available calcium indicators may be used. Examples of such indicators include Fluo-3, Fluo-3 AM, Fluo-4, Fluo-4 AM, Quin-2, Rhod-2, Rhod-2 AM (all manufactured by Dojindo Laboratories, Inc.), Fluo-8 AM, Cal-520 AM, and Rhod-4 AM (all manufactured by AAT Bioquest).

Fluorescence measurement of the calcium indicator in the cytoplasm is performed by standard methods. For example, when the calcium indicator is a fluorescent indicator, excitation light output from an excitation light source is irradiated onto a sample container containing cultured vascular smooth muscle cells, and the emitted fluorescence is detected over time by a fluorescence detector such as a fluorescence microscope, and a fluorescent image is taken. The average brightness is calculated from the captured image, and the amount of calcium ions in the cells can be quantified.

According to the screening method of the present invention, ingredients that are effective against dull skin caused by vasoconstriction can be efficiently screened.
Furthermore, it is possible to produce a skin dullness improving agent containing a skin dullness improving component screened by the screening method of the present invention.

The following describes the test examples and manufacturing examples, but the present invention is not limited to these.

Test Example 1 Screening of Inhibitory Components for Endothelin-Stimulated Increase in Calcium Ion Concentration in Vascular Smooth Muscle Cells Normal human aortic smooth muscle cells (HAMSCs) were cultured in Humedia-SG2 medium (Kuraray Industries, Inc.) at 37° C. for 6 days. The cultured HAMSCs were seeded on a 96-well glass plate and then cultured for 24 hours.
After incubation, the test substance was diluted to a concentration that did not cause cytotoxicity and added at 100 μL/well, and further incubated for 24 hours. After incubation, the medium was removed, and 5 μmol/L calcium indicator (Fluo-8, AAT Bioquest) dissolved in DMSO was diluted 1/1000 with HBBS medium, added at 100 μL/well, and incubated at 37° C. for 40 minutes.
The medium was removed and the cells were washed with HBBS medium, and 100 μL/well of fresh HBBS medium was added and incubated at 37° C. for 15 minutes. 1 nM endothelin-1 was added to the cells, and the change in intracellular calcium ion concentration due to calcium ion influx was measured for 2 minutes. The intracellular calcium ion concentration was quantified by fluorophotometric analysis, and the rate of increase (difference) in intracellular calcium ion concentration before and after the addition of endothelin-1 was evaluated. The results are shown in FIG. 1.

As shown in Figure 1, the sample to which endothelin-1 was added had about 1.8 times the amount of calcium ion influx compared to the sample to which endothelin-1 was not added. On the other hand, the sample to which Roman chamomile extract was added as a candidate material had significantly suppressed calcium ion influx, and the amount of calcium ion influx was less than that of the sample to which endothelin-1 was not added.
These results demonstrate that Roman chamomile extract is effective as an inhibitor of the increase in calcium ions in vascular smooth muscle cells due to endothelin-1 stimulation.

Preparation Example 1: Skin Lotion for Improving Dullness According to the formulation shown in Table 1, a skin lotion, which is the composition of the present invention, was prepared by a conventional method.

By continuously applying the dullness-improving lotion produced by the above method to the skin, it was found to have the effect of improving dullness.

The present invention can be applied to cosmetics, pharmaceuticals, etc.

Claims (4)

The composition contains an ingredient that inhibits an increase in calcium ion concentration in vascular smooth muscle cells due to endothelin stimulation, an ingredient that inhibits melanin production , and an ingredient that inhibits AGEs production ,
The component that inhibits an increase in calcium ion concentration in vascular smooth muscle cells due to endothelin stimulation is an extract of Roman chamomile (Chamaemelum nobile),
The melanin production inhibitor is a component selected from ascorbic acid and its derivatives,
The AGEs production inhibitor is a fermentation product of the calyx of a plant of the genus Hibiscus (Malvaceae) and lactic acid bacteria,
A dullness-improving agent for use against temporary dullness of the skin caused by vasoconstriction . The ascorbic acid and its derivatives are at least one selected from the group consisting of ascorbic acid, ascorbic acid glucoside, ascorbic acid phosphate, ascorbic acid sulfate, ascorbic acid fatty acid ester, and ascorbic acid alkyl ether;
2. The skin dullness improving agent according to claim 1, wherein the plant of the genus Hibiscus (Hibiscus L.) of the family Malvaceae is Hibiscus sabdariffa L. The skin dullness improving agent according to claim 1 or 2, wherein the content ratio of the melanin production inhibitor and the AGEs production inhibitor is, in dry mass, 0.01 to 10 mass parts of the melanin production inhibitor and 0.0001 to 0.008 mass parts of the AGEs production inhibitor, per 0.001 mass part of the inhibitor of the increase in calcium ion concentration in vascular smooth muscle cells due to endothelin stimulation . The skin dullness improving agent according to any one of claims 1 to 3 , which is used as a cosmetic.