KR20160001478A - Composition for Improving Acne - Google Patents

Abstract

Disclosed is a composition for improving acne, which uses a mixed extract of Curcuma longa Linnaeus roots, Magnolia Kobus barks, Sophora flavescens Aiton roots, and Centella asiatica (L.) Urbain.

Description

[Composition for Improving Acne]

The present invention relates to a composition for improving acne using ganoderma, Magnolia, Gossam, Centella asiatica, or a mixed extract thereof.

Acne is a polymorphic skin disease that occurs in the form of facial, chest and shoulder parts, pustular pustules, papules and nodules. It occurs mainly in adolescents with secondary maturity. It has been reported that about 80% of cases develop from 11 to 30 years of age and about 6% of cases occur after 30 years of age. Recently, however, the incidence of adult acne due to stress has been increasing. The disease can be uncomfortable to the patient and cause aesthetic damage due to scarring and skin pigmentation. As a result, interest in prevention and treatment of acne is increasing.

The exact cause of the occurrence of acne has not been elucidated and it has difficulties in proper treatment for various types of acne because it has diverse physiological and environmental aspects. (Farrar, MD and Ingham, E., Acne: Infertility, Fertilization, and Acne: A Case Report). In this study, Thus, as a basic method for treating acne, it is possible to reduce the activity of the sebaceous glands by correcting the trait changes due to follicular keratinization, thereby reducing the activity of the sebaceous glands. , Antimicrobial action against acne-induced major strains, and skin anti-inflammatory action.

Staphylococci and micrococci, which are aerobic bacteria, and bacteria belonging to the genus Propionibacterium, which are anaerobic bacteria, reside in normal skin, and acne may occur due to the activity of these strains. In particular, Propionibacterium acnes, an anaerobic strain with lipid affinity, has been identified as a major cause of acne (Burkhart CG et al., 1999, 75: 328). P. acnes hydrolyzes the triglycerides of sebum produced by androgen and other factors, and produces free fatty acids through the secretion of lipases and chemotactic factors, resulting in inflammatory responses . As the inflammatory reaction progresses, the hair follicles are destroyed and finally the inflammation reaction progresses to the dermis. Also, secondary infections by other bacteria can occur after inflammation.

One of the other causes of acne is the development of sebaceous glands, which occurs rapidly in the period of the Second Constitution. When the sebaceous gland is secreted by the development of the sebaceous glands, sebaceous glands accumulate in the hair follicles, causing the glands to rash. In addition, stagnant sebaceous follicles block the air circulation by blocking the hair follicle, which helps the growth of anaerobic bacteria such as P. acnes .

The follicular keratinization is easy to occur in the dermal papilla on the end of the hair follicle, and thick layers of skin are peeled off into the hair follicle, resulting in the closure of the dermal papilla, which forms the dermal patch. If the follicles or sebaceous glands are clogged or narrowed at the keratin skin, normal sebum discharge is inhibited and sebum becomes stagnant in the dermal papilla. As a result, the proliferation of the acne-related anaerobic bacteria is increased, and the production material stimulates the epithelial cells of the papilla, thereby exerting a keratinous effect. (Bae, Hyeon-Yeon, Sookmyung Women's University, Master Thesis, 2009. pp. 7).

To date, proposed methods of treating acne include the use of female hormones such as estrogen, which is a male hormone inhibitor, or antihistamines, to control sebaceous secretion, and the use of salicylic acid or a vitamin A derivative, retinoic acid, And anti-inflammatory drugs for inhibiting inflammation or antibiotic application methods for inhibiting bacterial infections are most commonly used. However, when conventional antibiotics such as tetracycline, erythromycin, and clindamycin are used, adverse effects such as the appearance of resistant bacteria, tissue, organ damage, and hypersensitivity are becoming serious problems. Therefore, it is necessary to study the antibacterial and anti-inflammation for the treatment of acne by using a naturally-occurring substance which is less toxic and can reduce side effects.

This is the background of the present invention.

It is an object of the present invention to provide a composition for improving acne.

Other and further objects of the present invention will be described below.

The inventors of the present invention have found that extracts of turmeric, Magnolia, Gossam, Centella asiatica, or a mixture thereof are extracted using 70% ethanol, and the extract of Propionibacterium acnes The antimicrobial activity of the mixed extracts was higher than that of the Magnoliae extracts which showed the antimicrobial activity. The antioxidant activity of the mixed extracts was also higher in the antioxidant activity. It was also confirmed that the mixed extract promoted the proliferation of human fibroblasts as well as inhibited the expression of COX-2, an anti-inflammatory agent.

The present invention is provided based on the results of such experiments and can be understood as an acne improvement composition comprising, as an active ingredient, a mixed extract of turmeric root, Magnolia bark, Gossam root and Centella asiatica on the one side, The composition of the present invention can be grasped as an antioxidant composition comprising an extract of Guanxi root, a extract of Magnolia bark, an extract of Guanxia root, an extract of Chenopodium ambrosioides L., or a mixed extract thereof. In another aspect, As an active ingredient can be identified.

As used herein, the term "extract" refers to a substance which is extracted with water, a lower alcohol having 1 to 4 carbon atoms such as methanol, ethanol, and butanol, methylene chloride, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, - Extracts obtained by leaching using dimethylformamide (DMF), dimethylsulfoxide (DMSO), 1,3-butylene glycol, propylene glycol or a mixed solvent thereof, and extracts obtained using supercritical extraction solvents such as carbon dioxide and pentane Extract means the fraction obtained by fractionating the extract or the extract. The extraction method can be applied by any method such as cold rolling, refluxing, heating, ultrasonic irradiation, supercritical extraction, etc. in consideration of the polarity, extraction degree, . The fraction extracted is a fraction obtained by suspending the crude extract in a specific solvent and mixing and leaving with a solvent having a different polarity. The crude extract is used in the order of increasing or decreasing the polarity, , And further includes fractions obtained by chromatography using properties such as size, charge, hydrophobicity, affinity, and the like. Also, the meaning of the extract includes concentrated liquid or solid extract from which the extraction solvent has been removed by freeze drying, vacuum drying, hot air drying, spray drying and the like.

In the present specification, the above-mentioned "active ingredient" means an ingredient that exhibits the desired activity alone or can exhibit activity together with a carrier that is itself inactive.

Terms not specifically defined in the present specification follow the meaning or the dictionary meaning commonly used in the art.

The composition of the present invention may be contained in any amount (effective amount) as long as it can exhibit an acne improving activity, an antioxidative activity or an anti-inflammatory activity depending on the purpose of use, formulation, compounding purpose and the like. And will be determined within the range of 0.001 wt% to 15 wt% based on the weight. Herein, the term "effective amount" refers to the amount of active ingredient which can exhibit an acne improving activity, an antioxidative activity or an anti-inflammatory activity in a mammal, preferably a human, to which it is applied. Such effective amounts can be determined experimentally within the ordinary skill of those skilled in the art.

The subject to which the composition of the present invention can be applied (prescription) is preferably a mammal and a person, particularly a human.

In a specific embodiment, the composition of the present invention can be identified as a cosmetic composition.

The cosmetic composition of the present invention may contain, in addition to the active ingredients thereof, conventional additives such as stabilizers, solubilizers, surfactants, vitamins, colorants and antioxidants, and carriers commonly used in cosmetic compositions .

The cosmetic composition of the present invention can be prepared into any of the formulations conventionally produced in the art and can be used as a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, , Oil, powder foundation, emulsion foundation, wax foundation and spray, 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.

When the formulation of the present invention is a paste, cream or gel, an animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as the carrier component .

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In the case of a spray, in particular, / Propane, < / RTI > butane or dimethyl ether.

When the formulation of the present invention is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component. Specifically, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid esters of sorbitan.

In the case where the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component is selected from aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters.

The cosmetic composition of the present invention can be prepared according to the production method of a cosmetic composition known in the art, including its effective component and the above-mentioned carrier component.

The composition of the present invention can be identified as a pharmaceutical composition in another specific embodiment.

The pharmaceutical composition of the present invention may be formulated into a wide variety of dosage forms, including pharmaceutically acceptable carriers, excipients and the like, in addition to the active ingredient, and may be formulated into a wide variety of dosage forms such as a localized preparation such as cream, lotion, ointment (semi-solid external drug), micro loo emulsion, (TTS) (e.g., patches, bandages, etc.), and the like.

The term "pharmaceutically acceptable" as used herein means that the application (prescribing) subject does not have the above-mentioned toxicity that is adaptable without inhibiting the activity of the active ingredient.

Examples of pharmaceutically acceptable carriers include lactose, glucose, sucrose, starch (e.g. corn starch, potato starch, etc.), cellulose, derivatives thereof (e.g. sodium carboxymethylcellulose, ethylcellulose, etc.), malt, gelatin, talc, (E.g., peanut oil, cottonseed oil, sesame oil, olive oil, etc.), polyols (e.g., propylene glycol, glycerin and the like), alginic acid, and the like. Depending on the formulation of the pharmaceutical composition of the present invention, one or more suitable ones may be selected and used. Suitable pharmaceutically acceptable carriers and formulations are described in Remington's Pharmaceutical Sciences (19th ed., 1995). The pharmaceutical composition of the present invention may further comprise an emulsifier (e.g., TWEENS), a wetting agent (e.g., sodium lauryl sulfate), a coloring agent, a flavoring agent, a stabilizer, a preservative, water, saline, a phosphate buffer solution and the like.

The excipient may be selected according to the formulation of the pharmaceutical composition of the present invention. For example, suspending agents such as sodium carboxymethyl cellulose, methyl cellulose, hydropropyl methyl cellulose, sodium alginate, polyvinyl pyrrolidone, .

The daily dose of the pharmaceutical composition of the present invention is usually 0.001 to 150 mg / kg body weight, and may be administered once or several times. However, since the dosage of the pharmaceutical composition of the present invention is determined in view of various related factors such as route of administration, age, sex, weight, and patient's severity of the patient, the dose is limited in any aspect to the scope of the present invention Should not be understood to be.

In a specific embodiment, the composition of the present invention can be identified as a food composition.

The food composition of the present invention may contain sweetening agents, flavoring agents, physiologically active ingredients, minerals and the like in addition to the active ingredients thereof.

Sweetening agents may be used in an amount that sweetens the food in a suitable manner, and may be natural or synthetic. Preferably, natural sweeteners are used. Examples of natural sweeteners include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose and maltose.

Flavors may be used to enhance taste or flavor, both natural and synthetic. Preferably, a natural one is used. When using natural ones, the purpose of nutritional fortification can be performed in addition to the flavor. Examples of natural flavoring agents include those obtained from apples, lemons, citrus fruits, grapes, strawberries, peaches, and the like, or those obtained from green tea leaves, Asiatica, Daegu, Cinnamon, Chrysanthemum leaves and Jasmine. Also, those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, banks and the like can be used. The natural flavoring agent may be a liquid concentrate or a solid form of extract. Synthetic flavors may be used depending on the case, and synthetic flavors such as esters, alcohols, aldehydes, terpenes and the like may be used.

Examples of the physiologically active substance include catechins such as catechin, epicatechin, gallocatechin and epigallocatechin, and vitamins such as retinol, ascorbic acid, tocopherol, calciferol, thiamine and riboflavin.

As the mineral, calcium, magnesium, chromium, cobalt, copper, fluoride, germanium, iodine, iron, lithium, magnesium, manganese, molybdenum, phosphorus, potassium, selenium, silicon, sodium, sulfur, vanadium and zinc can be used.

In addition, the food composition of the present invention may contain preservatives, emulsifiers, acidifiers, thickeners and the like as needed in addition to the above sweeteners.

Such preservatives, emulsifiers and the like are preferably added in a very small amount as long as they can attain an application to which they are added. The term " trace amount " means, when expressed numerically, in the range of 0.0005% by weight to about 0.5% by weight based on the total weight of the food composition.

Examples of the preservative which can be used include calcium sodium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate and EDTA (ethylenediaminetetraacetic acid).

Examples of the emulsifier which can be used include acacia gum, carboxymethyl cellulose, xanthan gum, pectin and the like.

Examples of the acidulant that can be used include acid, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, and phosphoric acid. Such an acidulant may be added so that the food composition has a proper acidity for the purpose of inhibiting the growth of microorganisms other than the purpose of enhancing the taste.

Agents that may be used include suspending agents, sedimentation agents, gel formers, bulking agents and the like.

In addition, herbal medicines may be added to improve flavor and palatability and to add other functionalities. Examples of medicinal herbs that can be added include mulberry extract, early-stage extract, antler extract, safflower extract, tosaja extract, , Gugija extract, licorice extract, Angelica gigantosa extract, Puerariae Radix extract, Ganoderma fragrant extract, Algerianthus sinensis extract, Acanthopanax root extract, Seed extract and the like.

As described above, the present invention can provide a composition for improving acne. The composition of the present invention can be commercialized as medicine, cosmetics or food.

Fig. 1 shows the results of the fibroblast proliferation promoting activity test.
Fig. 2 shows the results of the anti-inflammatory activity test.

Hereinafter, the present invention will be described with reference to Examples and Experimental Examples. However, the scope of the present invention is not limited to these examples and experimental examples.

< Example > Preparation of extract

Curcuma as an extraction raw material longa , root), Magnolia Kobus , bark), gosam ( Sophora flavescens , roots), Centella Asiatica , outposts) and mixtures of these equivalents were used.

The extracts of the extracts were filtered under reduced pressure and the filtrate was concentrated under reduced pressure at 70 ° C to obtain a solid extract.

< Experimental Example > Antioxidant activity, acne improvement activity, fibroblast proliferation promoting activity and anti-inflammatory activity experiment

< Experimental Example  1> Antioxidant activity - DPPH radical Scatters  Measure

(1) Production of DPPH

DPPH was added to 100% methanol so as to have a concentration of 0.1 mM to prepare a DPPH solution. Each of the samples of the examples was dissolved in 2% (w / v), 1% (w / v), 0.5% w / v) and 0.01% (w / v). After vortexing vigorously for the next 10 seconds and incubating in the dark for 30 minutes, the absorbance is measured at 517 nm using ELISA. Antioxidant activity was expressed as percentage of DPPH radical scavenging activity based on the absorbance of the control group using pure water.

The results are shown in [Table 1] below.

The results of the above Table 1 show that the mixed extracts of these extracts have higher antioxidative activity than the single extracts of turmeric, magnolia, gossam, and perilla. Especially, the mixed extract showed 80% higher antioxidant activity even at the low concentration of 0.1% (w / v).

< Experimental Example  2> Acne improvement activity-acne bacterium (P. acnes ) To measure antimicrobial activity against

&Lt; Experimental Example 2-1 > Experiment by disk diffusion method

The antimicrobial activity was measured by the Disc diffusion assay method. The test strain Acion bacteria ( Propionibacterium acnes ) were inoculated into 10 ml of liquid medium (GAM Broth) and subcultured for 3 hours at 37 ° C for 24 hours to be used as an antimicrobial activity test strain.

The antibacterial activity test was carried out by inserting a paper disc into a liquid medium (GAM Broth) in which the test strain was cultured, removing the paper disc, sufficiently wetting the paper disc with the liquid medium in which the test strain was cultured, The resulting solution was added to the paper disc in an amount of 20 μl each of the solution in a concentration of 1% (w / v), 1% (w / v), 0.5% (w / v) and 0.25% (w / v) Respectively. Then, the cells were incubated at 37 ° C for 24 hours and the size of the clear zone (mm) produced around the disc was measured to analyze the antimicrobial activity. As a negative control, distilled water used for the dilution of the extract was used.

The results are shown in Table 2 below.

[Table 2] also showed that the antibacterial activity of the mixed extract was better than that of the single extract.

<Experimental Example 2-2> The minimum inhibitory concentration ( Minimum inhibitory concentration , MIC )

The minimum inhibitory concentration was 5 ml of the liquid culture medium in a test tube, and the mixed extract sample of the example was 0.0625% (w / v), 0.03125% (w / v), 0.0156% w / v) and 0.0039% (w / v), and 5 ml of the solution was added to the test tube. The test strain cultured in the liquid medium ( Propionibacterium acnes ). The cells were then cultured at 37 ° C for 24 hours to determine the concentration at which the growth of the bacteria was no longer achieved.

As a result, the minimum inhibitory concentration was 0.0156%.

< Experimental Example  3> Fibroblast proliferation promoting activity experiment - MTT assay

Fibroblast cells were counted in a 96-well plate containing DMEM medium containing 0% FBS at a density of 1 × 10 ⁴ cells / well using a hemocytometer, and cultured for 48 hours. The samples of the following examples (mixed extracts) were dissolved in 0.1% (w / v), 0.05% (w / v), 0.01% (w / v), 0.005% (w / v), 0.001% And replaced with FBS-free DMEM for further incubation for 24 hours (sample dilution was diluted with culture medium). After the incubation, 50 μl of 3-, 4,5-di methylthiazol-2-yl) -2,5-diphenyl tetrazolium bromide solution (2.5 mg / ml) was added and the cells were further cultured for 3 hours. 100 μl of each solution was added to 96 wells, and the absorbance was measured at 570 nm using an enzyme-linked immunosorbent assay (ELISA).

The results are shown in Fig. 1 as a percentage of the untreated group.

Referring to the results of FIG. 1, fibroblasts proliferate in proportion to the treatment concentration.

< Experimental Example  4> Anti-inflammatory activity experiment - COX -2 expression effect measurement

<1> Cell culture

In this test, the level of expression of COX-2, an enzyme that induces inflammation in human keratinocyte line (HaCaT), was confirmed at the mRNA level. Specifically, the human keratinocyte line (HaCaT) was cultured in a 75 cm ² flask with Dulbecco's Modified Eagle's Medium (DMEM), 10% Fatal bovine serum (FBS) and 1% Antibiotic-Antimycotic (GIBCO, Cat No. 15240-062) And cultured at 37 ° C and 5% CO ₂ . When human keratinocytes were confluent at 80% confluence, cells were seeded at 110 ⁵ cells / well in a 6-well plate and cultured until cell confluence reached 80% confluence. After the medium was removed, the medium was replaced with a fresh medium containing a sample (mixed extract) at a concentration of 100 ppm, and further cultured for 48 hours under the cell culture conditions.

<2> RNA Isolation

RNA was isolated by total RNA isolation using QiAzol Lysis Reagent (QIAGEN). Cells from which the medium was removed were washed with PBS (phosphatized buffer saline), and 1 ml of QIAzol Lysis Reagent was added thereto, followed by treatment at room temperature for 10 minutes. After transferring to a new E-tube, 200 ul of chloroform (Amresco) was added, mixed well and incubated at room temperature for 5 minutes. Then, the mixture was centrifuged at 4 ° C and 12,000 rpm for 15 minutes to separate mRNA and protein DNA. 500 μl of the supernatant containing mRNA was transferred to a new E-tube, and 500 μl of Isopropanol (MERCK KGaA) was added thereto. The resulting mixture was inverted at room temperature for 4 to 5 times, and further reacted at room temperature for 7 minutes. And then centrifuged at 13,000 rpm for 30 minutes to precipitate the mRNA pellet. The supernatant was removed and the pellet was washed with 1 ml of 75% ethanol and dried. The RNA was dissolved in 20 ul of RNA Free Water (WelGENE Inc.) and the total RNA amount was adjusted to 5 μg / ml using a NanoDrop ^R Spectrophotometer (ND- ml.

&Lt; 3 > cDNA synthesis

cDNA synthesis was performed using PrimeScript ^™ reagent Kit (TaKaRa, RR037A). For details, 1ul of 50uM oligo-dT primer, 4u of Random Primer of 100uM, 4ul of 5X primeScript Buffer, and 1ul of PrimeScript RT enzyme Mix were added to 20ul of total RNA and reacted at 37 ℃ for 15 minutes to inactivate RTase And reacted at 85 ° C for 5 seconds.

<4> Real-time PCR

Real-time PCR was performed on the synthesized cDNA to analyze each gene. (GAPDH, COX-2) and a TaqMan Gene expression master mix (AB, Pro. No.4369016) were mixed with the cDNA, followed by 7500 Real time PCR system (applied biosystems). All genes were subjected to 60 cycles of 2 min at 50 ° C, 1 min at 95 ° C, and 15 sec at 95 ° C. Applied primers of Applied Biosystems (USA) were used for the primers used in the experiments. Expression rates are shown by calibrating each GAPDH.

<5> Result

The test was performed at 0.01% (w / v), 0.05% (w / v) and 0.1% (w / v) concentrations as shown in FIG. 2 and the expression of COX-2 2-fold. &Lt; / RTI &gt;

Claims (6)

The composition for improving acne according to the present invention is characterized in that the acne improving activity is due to the antibacterial activity against Propionibacterium acnes . / RTI &gt;
A composition for antioxidation comprising an effective component as an active ingredient, a mixture of extracts of rosemary root, extracts of Magnolia bark, extracts of rosemary root, extracts of rosemary extract, or roots of rosemary extract, magnolia bark,
A composition for antiinflammatory, comprising as an active ingredient, a mixed extract of Ganoderma root, Magnolia bark, Gossam root,
4. The method according to any one of claims 1 to 3,
Wherein the composition is a pharmaceutical composition.
4. The method according to any one of claims 1 to 3,
Wherein the composition is a cosmetic composition.
4. The method according to any one of claims 1 to 3,
Wherein the composition is a food composition.

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