KR20160093420A - Anti-dark circle composition comprising Berberine Hydrochloride or pharmaceutically acceptable salts thereof - Google Patents

Abstract

The present invention relates to a cosmetic composition comprising berberine or pharmaceutically acceptable salt thereof as an effective component for preventing or alleviating dark circles. More particularly, the composition of the present invention exhibits a strong effect of expressing heme oxygenase-1, and an effect of alleviating dark circles without side effects.

Description

The present invention relates to a cosmetic composition for improving dark circles comprising Berberine or a pharmaceutically acceptable salt thereof,

The present invention relates to a cosmetic composition for improving dark circles. More particularly, the present invention relates to a cosmetic composition which alleviates the dark circles that appear under the eyes due to dark circles and has a small side effect.

Heme oxygenase-1 is an enzyme that breaks heme, a by-product of hemoglobin decomposition, into biliverdin, etc., and performs decomposition and hydrolysis of hemoglobin in addition to enzymes such as biliverdin reductase and UGT1A1.

Dark circles occur in the skin under the eyes mainly due to dark shadow or pigmentation on the eyes, skin around the eyes, and the causes are various. The cause of dark circles can be classified into physical factors and biological factors. The physical factors of dark circles are the shadows caused by accumulation of abnormal fat in the fascia of the skin below the eyeball . In the case of biological factors, changes in hemoglobin due to accumulation of melanin and deposition of hemoglobin, which is caused by weakening of blood vessels, are considered to be prominently displayed on the skin near the eyeball having a relatively thin thickness relative to other skin.

The pathological effects of dark circles have not been proven, but many people, such as harming healthy images and negatively affecting their appearance, are ranked dark circles as a cosmetic problem of eyes. This dark circle phenomenon is recognized as an important problem to be solved for cosmetics.

At present, the way to treat dark circles exists in response to each cause. In the case of the pouch type skin tissue, which is a physical cause, the inhale or dispersion of the fat by the surgical operation or the removal of the tissue is possible, but the accessibility is not easy due to the trouble of the external activity due to the surgery and cost problems. Also, this surgical procedure can only be used to remove dark circles due to physical causes. In addition, the most commonly used way to treat dark circles in a cosmetic way is to mask the makeup cosmetics such as foundation or concealer. In addition, dark circles by melanin are similar to those of existing whitening cosmetics, but there is no countermeasure against deposition of hemoglobin decomposition materials.

Therefore, in the present invention, screening was performed on various natural material extracts in consideration of this situation, and a material for increasing the expression of Heme oxygenase-1, which promotes the hydrolysis of heme, a degradation by-product of hemoglobin, was selected.

SUMMARY OF THE INVENTION The present invention provides a cosmetic composition having excellent dark circles improvement effect.

In particular, the present invention provides a cosmetic composition using a natural herbal medicine extract, which has few side effects and is safe and has an excellent effect of improving dark circles.

The present invention provides a cosmetic composition for preventing and / or improving dark circles comprising berberine or a pharmaceutically acceptable salt thereof in order to solve the above technical problem.

The inventors of the present invention have experimentally found that berberine or a pharmaceutically acceptable salt thereof, preferably berberine hydrochloride, can alleviate the dark circles under the eyes and found that the dark circles of berberine or a pharmaceutically acceptable salt thereof Thereby providing a new use for improvement or prevention.

Further, the present invention increases the expression of Heme oxygenase-1, which promotes the hydrolysis of heme, which is a by-product of hemoglobin present in the skin when applied to the skin, thereby alleviating the dark circle symptoms occurring in the skin under the eyes and further preventing ≪ / RTI >

As used herein, the term "prevention" means to improve the flow of blood flow under the eyes and to increase darkening of the remaining heme in the skin to prevent occurrence of dark circles. Even the occurrence of a dark circle Is a term used in a broad sense to include.

As used herein, the term " improvement "refers to an improvement in the blood flow and relaxation of the under-eye area, so that the composition of the present invention is used before the composition of the present invention is used, Is a term used in a broad sense.

The formula for berberine hydrochloride of the present invention is C ₂₀ H ₁₈ ClNO ₄ , and its molecular weight is 371.81, which can be represented by the following formula (1).

[Chemical Formula 1]

The berberine according to the present invention may be chemically synthesized or extracted by a method known in the art, or a commercially available substance may be used.

The term "pharmaceutically acceptable salts " of the present invention refers to salts prepared with little or no toxicity or with acids or bases. When the compound of the present invention is relatively acidic, the base addition salts may be obtained by contacting a neutral form of such compound with a sufficient amount of the desired base and an appropriate inert solvent. Pharmaceutically acceptable base addition salts include, but are not limited to, salts formed with sodium, potassium, calcium, ammonium, magnesium or organic amino. Where the compounds of the present invention are relatively basic, acid addition salts can be obtained by contacting neutral forms of such compounds with a sufficient amount of the desired acid and a suitable inert solvent. Pharmaceutically acceptable acid addition salts include those derived from organic acids such as propionic acid, isobutyric acid, oxalic acid, malic acid, malonic acid, benzoic acid, succinic acid, sueric, fumaric acid, mandelic acid, phthalic acid, benzenesulfonic acid, Phosphoric acid, monohydrogencarbonic acid, phosphoric acid, monohydrogenphosphoric acid, dihydrogenphosphoric acid, sulfuric acid, monohydrogensulfuric acid, hydrogen iodide, phosphorous acid, And the like, but are not limited thereto. But are not limited to, salts of amino acids such as arginate and analogs of organic acids such as glucuronic or galactunoric acids. Preferably, the pharmaceutically acceptable salt of berberine of the present invention may be a berberine hydrochloride.

The berberine of the present invention may exist in solvated as well as unsolvated forms, including forms such as hydrates, ethanolates and the like. The berberine of the present invention may exist in crystalline or amorphous form, and all such physical forms are included within the scope of the present invention.

The inventors of the present invention have evaluated the expression ability of Heme oxygenase-1 enzyme which improves dark circles by using berberine. As a result, berberine and its pharmaceutically acceptable salts showed strong Heme oxygenase-1 expression.

The cosmetic composition of the present invention may contain 0.0005 to 10% by weight, preferably 0.001 to 5% by weight, based on the total weight of the composition, of berberine or a pharmaceutically acceptable salt thereof.

If the content of berberine or its pharmaceutically acceptable salt is less than 0.0005% by weight, it is difficult to expect the expression of Heme oxygenase-1, and if it exceeds 10% by weight, no marked effect is expected as an increasing content.

The cosmetic composition comprising berberine or a pharmaceutically acceptable salt thereof of the present invention can be prepared as a cosmetic composition according to a conventional method for producing a cosmetic composition.

In addition, the cosmetic composition according to the present invention may contain components commonly used in addition to the above-mentioned berberine, and examples thereof include moisturizers, thickeners, surfactants, oil bases, preservatives, antioxidants, alcohol, fragrances, pH regulators, Of course, a chemical component for doubling the dark circles improvement effect by the berberine can be blended in an appropriate amount.

The cosmetic composition according to the present invention may be prepared in any form conventionally produced in the art, and may be in the form of solution, suspension, emulsion, paste, gel, cream, lotion, soap, surfactant- , But is not limited thereto. More specifically, it can be manufactured in the form of a soft lotion, a nutritional lotion, a nutritional cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder.

The preferred formulations of the present invention are lotions, creams, and essence formulations in which the ingredients are uniformly dispersed in the cosmetic composition and the effect of the present invention is best exhibited.

Compositions containing berberine or a pharmaceutically acceptable salt thereof of the present invention may have an effect on the prevention and improvement of dark circles.

In particular, the composition containing berberine or a pharmaceutically acceptable salt thereof according to the present invention is excellent in the improvement of hemoglobin by hemoglobin in the symptoms of dark circles and the pigment deposition phenomenon.

The composition of the present invention was found to be superior in clinical improvement of under-eye brightness.

The composition of the present invention is excellent in prevention and improvement of dark circles while having few side effects.

1 is a schematic diagram of a vector for expression of luciferase comprising a promoter region (-890 to +80) of Heme oxygenase-1 (HO-1).
FIG. 2 is a graph showing luciferase activity measured to examine the expression level of Heme oxygenase-1 according to the concentration of berberine hydrochloride.
FIG. 3 is a graph showing the mRNA expression level of Berberine hydrochloride-treated experimental group by quantifying the mRNA expression level of the negative control group at 1.0.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the embodiments according to the present invention can be modified into various other forms, and the scope of the present invention should not be construed as being limited to the embodiments described below. The embodiments of the present invention are provided by way of example to facilitate a specific understanding of the present invention.

The berberine hydrochloride used in the following examples of the present invention was purchased from Sigma.

Example 1 Evaluation of Heme oxygenase-1 transcription activation

Similar to human skin cells, Hacat, a cell line expressing Heme oxygenase-1, was subcultured in Dulbecco's modified Eagle's medium (DMEM; Invitrogen, USA) containing steroid hormone-depleted fetal bovine serum (5% charcoal stripped FBS) . A reporter vector was constructed to examine the promoter activity of Heme oxygenase-1. For this, the promoter region (-890 ~ + 80) of Heme oxygenase-1 from human genomic DNA was amplified by PCR, and the primers used at this time are shown in Table 1 below.

Sequence Forward CGGGGTACCGTGTCCCACGCATTCCAGCA Reverse CATGCCATGGTATGCTCGGCGGGTCACGTG

The amplified DNA was cut using KpnI and NcoI restriction enzymes and cloned into a pGL3 vector expressing firefly luciferase, which vector is shown in Fig.

The vector was injected into Hacat cells via transfection.

The prepared plasmid vector and a plasmid always expressing Renilla luciferase were used in a promoter assay in combination with a DNA mixture in a ratio of 40: 1. 2 x 10 ⁵ hacat cells were plated on a 24 well plate and cultured for 18 hours. Then, 1 μg of the plasmid gene, Heme oxygenase-1 reporter, was analyzed using Lipofectamine® Transfection reagent (Invitrogen, USA) Transient transfection was performed according to the manufacturer's instructions. The cells were incubated for 24 hours, washed with phosphate buffered saline (PBS), and then added with 10 ppm of berberine hydrochloride in DMEM without fetal bovine serum. The cells were treated with the same amount of control as the control (control), cultured for 24 hours, washed with phosphate buffer, and the cells were lysed using a cell lysis buffer. Then, the supernatant was extracted using a centrifuge to obtain luciferase assay, The luminescence was measured with an Elmer Victor 3 (Perkin Elmer Victor 3, USA) and is shown in FIG.

As shown in FIG. 2, the berberine hydrochloride according to the present invention increases the expression of Heme oxygenase-1 to a significant extent, indicating high firefly luciferase activity. The measurements corrected firefly luciferase levels to renilla luciferase levels.

<Example 2> Modulation effect of Heme oxygenase-1 gene expression

Hacat cells were cultured in a biotechnology institute, and Dermal basal cell media (DBCM, ATCC, USA) was used. 8 x 10 ⁵ cells were seeded on 6-well plates and cultured for 18 hours, then treated with 10 ppm of berberine hydrochloride for 24 hours. Cells were harvested and washed with PBS. Total RNA was extracted with RNeasy mini kit (Qiagen, Germany), and 1 μg of RNA was quantitatively analyzed using a GeneAmp ^® RNA PCR kit (Applied Biosystems, USA) reverse transcription). Reverse transcription was performed using a Mycycler ^® PCR instrument (Biorad, USA).

Quantitative real time PCR was performed using Heme oxygenase-1 and GAPDH Taqman ^® probe (Invitrogen, USA, Hs01110250_m1, Hs03929097_g1). Real-time PCR was performed using a CFX96 Touch ™ Real-Time PCR Detection System instrument (Biorad, USA). Experimental results obtained by real-time PCR were compared with the housekeeping gene Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) using the ΔCt method (Livak KJ, Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2 -Delta Delta C (T)) Method, Methods, 25, 402-408 (2001)). In addition, mRNA expression level of the test group treated with berberine hydrochloride was quantified based on the mRNA expression level of the negative control group as 1.0, and it is shown in Fig.

&Lt; Preparation Example >

Composition (% by weight) Manufacturing example Berberine hydrochloride One Stearic acid 15 Cetanol One Potassium hydroxide 0.7 glycerin 5 Propylene glycol 3 antiseptic Suitable amount incense Suitable amount Purified water to 100

As shown in Table 3, cream containing berberine hydrochloride as an active ingredient was prepared by a conventional method.

&Lt; Experimental Example 1 >

Experiments were conducted to measure the effect of improving the lightness difference of the skin under the eyes using the product manufactured in the following manner by the following method. The effect test was carried out by using the cream of the above-mentioned production example as an example, and in the examples, berberine hydrochloride This study was carried out with 30 participants using the cream which did not contain the cream as a comparative example. Participants of the experiment were periodically applied the cream of the example and the comparative example to the area under the left and right eyes of each participant for one week in the morning, one time before bedtime, two times a day for four weeks. After 4 weeks, the subject was sufficiently stabilized for 10 minutes in a constant temperature and humidity room, and then the change in L ^* , a ^* , and b ^* values of the pigment deposit under the eyes was measured using Chromameter CR-400 (Minolta, Japan) The ratio of the number of persons showing a significant difference in each figure was confirmed. L * is the lightness axis, a * is the green / red axis, and b * is the blue / yellow axis. The results of dark circles improvement are shown in Table 4.

Comparative Example (Below left eye) Example (under the right eye) L ^* 9 (30.0%) 22 (73.3%) a ^* 12 (40.0%) 19 (63.3%) b ^* 13 (43.3%) 20 (66.7%)

Table 4 shows the results of measuring changes in L ^* , a ^* , and b ^* values at the sites where pigment under eyes are deposited. According to the results, it can be seen that the cosmetic composition containing berberine hydrochloride of the present invention is effective for improving the brightness difference under the eyes as compared with the comparative example not containing berberine hydrochloride, and thus is effective for improving dark circles under eyes I think.

Claims (5)

A cosmetic composition for prevention or improvement of dark circles comprising berberine or a pharmaceutically acceptable salt thereof as an active ingredient. The cosmetic composition according to claim 1, wherein the pharmaceutically acceptable salt thereof is berberine hydrochloride. The cosmetic composition according to claim 1, wherein the berberine or a pharmaceutically acceptable salt thereof is contained in an amount of 0.0005 to 10% by weight based on the total weight of the composition. 2. The cosmetic composition according to claim 1, wherein the berberine or a pharmaceutically acceptable salt thereof increases the expression of Heme oxygenase-1. A composition for the expression of Heme oxygenase-1 comprising berberine or a pharmaceutically acceptable salt thereof as an active ingredient.

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