JP2019011253A - Epidermal thickening inhibitor - Google Patents

Abstract

To provide an epidermal thickening inhibitor that inhibits epidermal thickening caused by ultraviolet exposure, and a cosmetic preparation, quasi drug, pharmaceutical or food and drink containing the epidermal thickening inhibitor.SOLUTION: The present invention provides an epidermal thickening inhibitor that inhibits epidermal thickening caused by ultraviolet exposure, containing Areca catechu L. (Palmae) seed extract as an active ingredient, and a cosmetic preparation, quasi drug, pharmaceutical or food and drink containing the epidermal thickening inhibitor. The cosmetic preparation is preferably in the form of skin application such as a lotion, cream, latex, cosmetic liquid.SELECTED DRAWING: None

Description

  The present invention relates to an epidermal thickening inhibitor.

  Epidermal thickening is one of the skin abnormalities caused by chronic UV exposure on the face and back of hands. On the other hand, it is known that an epidermis layer of an unexposed part such as a buttock gradually contracts with aging (see Non-Patent Document 1).

  Thickening of the epidermis is one of the causes of lowering the quality of life (QOL), especially for women, because it involves not only the effects on the human body in pathological aspects but also cosmetic losses such as rough texture. it is conceivable that.

  Substances that are said to be involved in the development of epidermis thickening include cytokines such as EGF (epidermal growth factor), FGF-2 (fibroblast growth factor-2), tryptase, interleukin-22, fibronectin, ADAM ( A biological substance such as A disintegrin and metalloprotease) can be mentioned, but the entire mechanism of the onset mechanism has not yet been elucidated (see Non-Patent Document 2 and Patent Documents 1, 2, and 3).

  Epidermis thickening is a phenomenon that the epidermis layer becomes thicker, so in the hope of returning the thickened epidermis to a normal state, promoting epidermis thickening containing retinoids and tocopherol that has the effect of promoting epidermis turnover An agent and a cosmetic preparation containing α-hydroxycarboxylic acid having a peeling effect have been proposed (see Patent Documents 4 and 5).

  However, the above-mentioned epidermal thickening promoter and cosmetic preparation are applied for the purpose of treating symptoms caused by exposure to ultraviolet rays, and not for the purpose of preventing epidermal thickening in advance.

  In addition, for the purpose of suppressing epidermis thickening caused by exposure to ultraviolet rays, an epidermis thickening inhibitor composed of sphingosine is known (see Patent Document 6). In addition to difficulty in ensuring stability over time, there are many restrictions on industrial applicability due to the limited types and amounts of oils and surfactants that can be blended from the viewpoint of safety. It was a problem (see Patent Documents 7, 8, and 9).

  By the way, a tryptase activity inhibitor containing a betel nut seed extract or the like as an active ingredient has been proposed as a drug using the betel nut seed extract (see Patent Document 10). However, there has been no report that the tryptase activity inhibitor has been confirmed to substantially suppress epidermal thickening in the betel nut seed extract.

JP 2012-158573 A JP 2013-129617 A JP 2006-137744 A JP 2013-209334 A Japanese Patent Laying-Open No. 2015-147797 Japanese Patent No. 3342672 JP 2009-298757 A JP 2006-084360 A Japanese Patent No. 5112673 JP 2007-186457 A

Standard Dermatology 8th pp. 147-148 (Medical School) Hideki Fujita Journal of Japanese Society for Clinical Immunology (Vol.35 No.3) pp. 168-175

  Therefore, the present invention provides an epidermis thickening inhibitor that suppresses the thickening of skin caused by exposure to ultraviolet rays, characterized in that it contains a betel nut seed extract as an active ingredient, and the problem to be solved by the invention To do.

  As a result of intensive studies aimed at solving the above-mentioned problems, the present inventors have found that the betel nut tree seed extract has an action of suppressing epidermis thickening caused by ultraviolet exposure.

Accordingly, the present invention relates to the following.
(1) An epidermis thickening inhibitor that suppresses epidermis thickening caused by exposure to ultraviolet rays, comprising a betel nut extract as an active ingredient.
(2) Cosmetics, quasi-drugs, pharmaceuticals, or food / beverage products containing the epidermis thickening inhibitor as described in said (1).

  An epidermis thickening inhibitor that suppresses epidermis thickening caused by ultraviolet rays, characterized by containing the betel nut seed extract of the present invention, is widely used in various forms as cosmetics, quasi drugs, pharmaceuticals, or foods and drinks. Can be used universally.

  The details of the present invention will be described below, but the technical scope of the present invention is not limited only to the matters of this item, and the present invention can take various forms as long as the object is achieved.

  In order to reach the present invention, the present inventors have clarified the phenomenon of epidermal thickening caused by ultraviolet exposure as follows. That is, when the epidermis layer is exposed to ultraviolet rays, depending on the conditions such as the exposure amount, epidermal cells can repair and re-synthesize damage received by biomolecules such as nucleic acids, proteins and lipids, and reactive oxygen species (ROS). : Reactive oxygen species) and the secretion of cytokines, etc., and showed that the surrounding environment can be changed. Although the effects of UV exposure also lead to cell death (apoptosis, necrosis, etc.), cell death in the epidermis layer represents a decrease in epidermal cells, and in view of the fact that epidermal atrophy is caused. It has been clarified that the cause of the thickening of the epidermis as shown in Non-Patent Document 1 is due to the proliferation of cells themselves and surrounding cells that did not cause cell death.

  Based on the above facts, the inventors considered that if cell proliferation caused by UV exposure can be suppressed, epidermal thickening can be suppressed, and as a result of intensive studies, among the various components studied It came to discover that a betel nut tree seed extract has the effect | action which suppresses epidermal thickening.

  The betel tree seed extract used in the present invention has the scientific name Areca Catchu L. It is obtained from Palmae betel nut (large rind, palm) and its related plants.

  The method for adjusting the betel nut tree seed extract is not particularly limited. Can be mentioned.

  As the extraction solvent, generally, water, lower monohydric alcohols (methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, etc.), liquid polyhydric alcohol (propylene glycol, 1, 3) -Butylene glycol, glycerin, etc.), lower alkyl esters (ethyl acetate, etc.), hydrocarbons (benzene, hexane, pentane, etc.), ketones (acetone, methyl ethyl ketone, etc.), ethers (diethyl ether, tetrahydrofuran, dipropyl ether, etc.) And acetonitrile.

  Examples of commercially available products include Areca Cache Extract (Bioland).

  The content of the betel nut tree seed extract is not particularly limited as long as it has an effect of suppressing epidermis thickening, but is, for example, 0.001 to 1% by mass with respect to the total amount of the composition, 0.001 -0.5 mass% is preferable, and 0.001-0.1 mass% is further more preferable. In addition, these values are content as dry matter.

  The epidermal thickening inhibitor containing the betel nut seed extract can be used as a cosmetic, a quasi-drug, a pharmaceutical or a food or drink as it is or with other components.

  Specific examples of cosmetics and medicinal cosmetics include, but are not limited to, skin care cosmetics, makeup cosmetics, fragrance cosmetics, body care cosmetics, hair care cosmetics, and the like.

  Specific forms of pharmaceuticals and quasi-drugs are not particularly limited. However, as long as they are applied to the skin, in addition to the above-described specific forms of cosmetics and medicinal cosmetics, for example, anti-smell agents, garlic powders , Hair restorer (hair restorer), hair remover, hair dye (depigmenting agent, decoloring agent), permanent wave agent, bath agent, repellent, etc. More specific dosage forms include ointments, creams Etc. Examples of forms not applied to the skin include injections (muscular, subcutaneous, intradermal), infusions, oral preparations, nasal drops and the like.

  The inhibitor and the composition containing the inhibitor of the present invention are not particularly limited with respect to the production method, and can be produced by a method generally known to those skilled in the art.

  EXAMPLES Hereinafter, although an Example demonstrates this invention further more concretely, this invention is not limited to these Examples.

[1. Preparation]
Areca tree seed extract (Areca Catalyst Extract (Bioland) powdered) was pre-dissolved in dimethyl sulfoxide (Dimethylsulfoxide) and then 5% (v / v) fetal bovine serum (FBS; Life technologies) Was dissolved in Dulbecco's modified Eagle medium (DMEM; Life technologies) to a final concentration of 50 μg / mL, and this was used as a test solution. The final concentration of DMSO was 0.1% (v / v). The rice extract was dissolved in DMEM containing 5% (v / v) FBS to a final concentration of 50 μg / mL without using DMSO, and this was used as a test solution.

[2. Culture of immortalized human epidermal cell line HaCaT cells and irradiation with ultraviolet rays]
HaCaT cells were suspended using DMEM containing 10% (v / v) FBS, and seeded in a 35 mm petri dish at 9.0 × 10 ⁵ cells. Thereafter, the 35 mm petri dish was cultured for 24 hours under the conditions of 37 ° C. and CO ₂ concentration of 5%.

0.1 mL of the test solution prepared above was added to each petri dish from which the culture supernatant after culture was removed, and further cultured for 24 hours. Each petri dish from which the culture supernatant after the culture was removed was washed twice with 2 mL of Hanks balanced salt solution (HBSS; Nissui Pharmaceutical), and 2 mL of DMEM without phenol red was added to each petri dish. Then, it irradiated with the ultraviolet-ray and culture | cultivated for 24 hours. The irradiation intensity of ultraviolet A wave (UVA) was 0 (no irradiation) and 10 J / cm ^2, and the irradiation intensity of ultraviolet B wave (UVB) was 0 (no irradiation) and 25 mJ / cm ² . (FL20S-BLB; Toshiba Corporation) was used as the UVA lamp, and (TL20W / 12RS, UVB Broadband; Philips) was used as the UVB lamp. For the intensity measurement, a digital UVX ultraviolet intensity meter UVX Radiometer (UVP) and a UV irradiation intensity sensor UVX-31 (UVP) were used.

On the same day when the HaCaT cells in each dish were irradiated with UVA or UVB, the HaCaT cells were suspended using DMEM containing 10% (v / v) FBS, and 3.5 × 10 ⁴ cells / well in a 96-well culture plate. Sowing. Thereafter, the 96-well culture plate was cultured under the conditions of 37 ° C. and CO ₂ concentration of 5% for 24 hours.

  The culture supernatant in the 96-well culture plate after the culture was removed and washed twice with 2 mL of HBSS. Thereafter, 0.1 mL of the culture supernatant obtained from each petri dish irradiated with UVA or UVB was added to each well and cultured for 24 hours. The culture supernatant obtained from each petri dish irradiated with UVA or UVB was centrifuged at room temperature and a speed of 3,000 rpm for 5 minutes to remove unnecessary substances before being added to each well. The culture supernatant in the 96-well culture plate after the culture was removed and washed twice with 2 mL of HBSS. For each well after culture, protein quantification described below was performed. The number of wells required to acquire one data was 7 or more.

[3. Protein determination (measurement of cell growth rate)]
0.05 mL of a phosphate buffer solution containing 0.5% (v / v) Triton X-100 was added to each well of a 96-well culture plate to completely lyse the cells. To each well, 0.05 mL of a solution prepared by mixing a ratio of Reagent A and Reagent B of Pierce BCA Protein Assay Kit (Thermo Fisher Scientific) at a ratio of 50: 1 was added to each well and mixed on a shaker for 1 minute. Then, it incubated at 37 degreeC for 10 minute (s), and the light absorbency in 550 nm was measured using the light absorption microplate reader Vmax (Molecular Device).

[4. analysis method]
The protein amount was calculated from the obtained absorbance based on the calibration curve. Bovine serum albumin (BSA; Sigma Aldrich) was used to prepare a calibration curve. The BSA concentration was 2, 1, 0.5, 0.25, 0.125, and 5 mg / mL. In the result (1) described later, the cell growth rate when UVA or UVB is not irradiated is controlled 1, and in the result (2) described later, the cell growth rate when there is no sample and UVA or UVB irradiation is controlled 2. Control 3 was used. Using these as controls (cell growth rate = 100%), the cell growth rate when each test solution was allowed to act was calculated. Using statistical analysis described later, it was determined that there was an effect of suppressing epidermal thickening when cell proliferation was significantly suppressed.

[5. Statistical analysis]
For the statistical significance test in the test, Student's t-test was used, and the test was performed under unpaired two-sided test conditions. The significance level was 5% (p <0.05).

[6. Result (1)]
First, as a result of examining whether cell growth is induced by the effect of irradiation with UVA or UVB, as shown in Table 1 below, UVA or UVB irradiation was compared with non-UVA or UVB non-irradiation (control 1). As a result, it was confirmed that the cell proliferation rate was significantly increased.

[6. Result (2)]
Next, as a result of calculating the cell proliferation rate under UVA or UVB irradiation at the time of adding the test solution, as shown in Table 2 described later, compared with UVA or UVB non-irradiation (controls 2 and 3), Example 1 Significantly suppressed cell proliferation induced by UVA or UVB irradiation effects. However, Comparative Example 1 did not show a significant cell growth inhibitory effect. It should be noted that the use of DMSO as a solvent had no effect on this test system, and no toxicity of the test solution used to cultured cells was observed. As Tables 1 and 2 show, the test solution of Example 1 significantly suppressed cell growth due to the influence of UVA or UVB irradiation.

  Various formulation examples of the present invention are given below, but the technical scope of the present invention is not limited to these formulation examples.

(Prescription Examples 1 to 4) Lotion

(Prescription Examples 5-8) Cream

(Prescription Examples 9-12) Emulsion

(Prescription Examples 13 to 16) Essence

  Since the present invention can suppress the thickening of the epidermis, it is expected to reduce adverse effects on the human body based on the thickening of the epidermis.

Claims (2)

  An epidermis thickening inhibitor that suppresses thickening of skin caused by exposure to ultraviolet rays, comprising a betel nut seed extract as an active ingredient.   Cosmetics, quasi-drugs, pharmaceuticals or foods and drinks containing the epidermis thickening inhibitor according to claim 1.