KR101382137B1 - A composition comprising the complex extract of corylopsis gotoana and erythronium japonicum - Google Patents

Abstract

The present invention relates to a cosmetic composition for antioxidation and anti-inflammation and a pharmaceutical composition for preventing or treating atopic dermatitis. containing a mixed extract of Corylopsis gotoana and Erythronium japonicum. The mixed extract of Corylopsis gotoana and Erythronium japomicum of the present invention is a natural material and has an excellent antioxidation and anti-inflammatory effects without cytotoxicity, and alleviatesskin itching, a rash, and pruritus in a patient with atopic dermatitis, thereby being used as a functional cosmetic material alleviating skin oxidation and skin inflammation or a cosmetic material and a therapeutic agent for alleviating atopic dermatitis. [Reference numerals] (AA) Before applying; (BB) After applying

Description

A composition comprising the complex extract of Corylopsis gotoana and Erythronium japonicum

The present invention relates to an antioxidant and anti-inflammatory cosmetic composition and a pharmaceutical composition for the prevention or treatment of atopic diseases comprising a mixed extract of hyacinth and ulji.

In general, skin irritant substances are known to exist in daily life, and skin-related products often contain substances that can cause irritation, inflammation, and allergies. In addition, more than 10% of people who use personal hygiene and skin care products complain of skin side effects, and in the US in the 1990s, more than 50% of consumers thought they were sensitive skin. . Recently, the proportion of sensitive skin is gradually increasing due to factors such as physical and chemical substances such as UV, smoking, pollutants, heavy metals, drying of external environment living spaces, allergies of individuals, and stress. When such physical and chemical external environment causes abnormal cell action or excessive production of active oxygen to the extent exceeding the capacity of the defense system, oxidative stress is caused, and skin irritation and aging are promoted. In order to protect the skin from such various stimuli, various functional cosmetics as well as pharmaceuticals have been developed.

Various ingredients are used to make cosmetics, and typically 20 to 50 types of ingredients are used. Most of the ingredients used in the manufacture of cosmetics have been tested for their safety through skin irritation tests, but there have been reports of irritation such as inflammation, itching and allergies when applied to actual products.

If skin irritation is aggravated, it appears as an inflammatory response to the skin or in the form of allergic or atopic skin. Inflammation is inherently a series of defenses aimed at minimizing the response and restoring the damaged area when cells or tissues are damaged by any cause, and cause dysfunction due to pain, edema, redness, and fever. These inflammations return to normal after the initial state in the normal state, but when stimulants stimulating the inflammation do not disappear, or when the inflammatory reaction continues, various side effects such as cell damage occur. There are various substances for anti-inflammatory drugs, but in order to use them in cosmetics and medicines, they must be anti-inflammatory substances with no side effects in terms of skin safety.

Various methods have been tried to alleviate skin irritation, for example, to remove substances causing skin irritation. However, many cases have been reported that the functional material itself acts as an irritant. For example, Lactic acid, Glycolic acid, Salicycic acid, Retinoid, etc. are contained in cosmetics to promote skin cell regeneration or anti-wrinkle effects but cause skin irritation. . Accordingly, in recent years, many studies have been conducted to relieve stimulation by neutralizing acidity by adding strong alkali such as NaOH or KOH to stimulus-inducing functional raw materials, but such a prescription has a problem of lowering the efficacy of existing raw materials.

Therefore, in order to solve the above problems, research on cosmetics based on natural friendly substances having excellent effects while minimizing irritation to skin is increasing.

Earl is a perennial plant of the Liliaceae family, with long oval leaves and dark branched spots. When the spring snow melts, leaves sprout and beautiful flowers of eggplants bloom. Take the ginseng in spring or summer and dry it in the sun or use it as a raw medicine. Young leaves are dried and eaten as herbs. In oriental medicine, Rhizome was used as a nourishing tonic instead of Sanjago, and in the private sector, it was used to treat strong heart, fever, detoxification, swelling, and ulcers. [Park Jong Hee, Korea Herbal Medicine Book, Book Publishing Company, 2004].

The creeper is a plant of the Hamamelidaceae and grows in the deep mountains of the southern and central regions of Korea and at the foot of the mountain below 900 meters above sea level. Heary is a deciduous shrub about 4m high and has nicknames such as pine luminous print, lead print, and lead wood. The root skin of diary is used as a herbal medicine called lead print, and it was prescribed for cold, body aches, chills and fever [Park Jong Hee, Korea Herbal Medicine Book, Shinil Publishing Co., 2004].

It is an object of the present invention to provide an anti-inflammatory and anti-inflammatory cosmetic composition comprising a mixed extract of hyacinth and scabies.

It is also an object of the present invention to provide a cosmetic composition for improving atopy disease, including a mixed extract of hyacinth and ulji and a pharmaceutical composition for preventing or treating atopy disease.

The present inventors, while diligently researching natural products exhibiting anti-inflammatory and anti-inflammatory effects, and atopy-improving effect, the mixed extracts of hyacinth and scab are not only harmless to the human body but also exhibit excellent effects, The present invention was completed by confirming that the extracted mixed extract exhibits a particularly excellent effect.

The present invention provides an anti-inflammatory and anti-inflammatory cosmetic composition comprising a mixed extract of hyacinth and scabies.

In another aspect, the present invention provides a cosmetic composition, wherein the mixed extract is extracted using water or alcohol as an extraction solvent.

The present invention also provides a cosmetic composition, wherein the alcohol is a lower alcohol having 1 to 4 carbon atoms.

The present invention also provides a cosmetic composition, wherein the alcohol is 30% (v / v%) or 70% (v / v%) ethanol.

In another aspect, the present invention provides a cosmetic composition, which contains a diary and rust in a weight ratio of 1: 0.1 to 1: 3.

In another aspect, the present invention contains a mixed extract extracted from 1: 1 to 1: 3 weight ratio of hydroxy and scab, 30% (v / v%) or 70% (v / v%) ethanol as the extraction solvent Provides a cosmetic composition for anti-oxidant and anti-inflammatory.

In another aspect, the present invention provides a cosmetic composition for improving atopic diseases, including a mixed extract of hyacinth and earlyge.

In another aspect, the present invention provides a pharmaceutical composition for preventing or treating atopy disease, including a mixed extract of hyacinth and ulji.

The mixed extract of the hyacinth and ulji of the present invention is a natural substance, there is no cytotoxicity, has excellent antioxidant and anti-inflammatory effects, and can improve skin itching, erythema, dry skin, etc. in patients with atopic diseases, thereby preventing skin oxidation and skin inflammatory reactions. It can be usefully used as a functional cosmetic to improve or a cosmetic and a therapeutic agent for atopic disease improvement.

1 is a diagram showing the results obtained by comparing the skin condition before and after applying the cream containing the mixed extract of Example 2 to the skin of atopic patients.

The present invention provides an anti-inflammatory and anti-inflammatory cosmetic composition comprising a mixed extract of hyacinth and scabies.

The mixed extract of the hyacinth and ulge of the present invention is not cytotoxic and has excellent antioxidant and anti-inflammatory effects and can improve skin itch, erythema, dry skin and the like in atopic dermatitis patients.

The mixed extract may be water, alcohol, ethyl acetate, glycerin, ethylene glycol, propylene glycol, butylene glycol and mixed solvents thereof (for example, hydrous alcohol, hydrous glycerin, hydrous ethylene glycol, hydrous propylene glycol, hydrous butylene glycol It may be selected from), it is preferably extracted with a solvent selected from water, anhydrous alcohol having 1 to 4 carbon atoms or hydrous alcohol. Here, the alcohol having 1 to 4 carbon atoms may be methanol, ethanol, n-propanol, iso-propanol, n-butanol, iso-butanol or tert-butanol. Ethanol is preferred.

The ethanol may be 10 to 100% (v / v%) ethanol, preferably 30 or 70% (v / v%) ethanol aqueous solution (hereinafter referred to as% ethanol).

The mixed extract may be an extract extracted by mixing the hyacinth and the ratio in a weight ratio of 1: 0.1 to 1: 3, preferably a mixed extract extracted by mixing in a weight ratio of 1: 0.3 to 1: 3, more preferably 1 It is a mixed extract extracted by mixing in a weight ratio of 1 to 1: 3.

The mixed extract is preferably mixed with 30% or 70% ethanol by mixing the ratio and weight ratio 1: 1: 1 to 1: 3.

The mixed extract of the present invention may include not only an extract obtained by mixing hyacinth and ulge, but also a composition mixed after extracting each constituent of hyacinth and ulzz.

The mixed extract of the hyacinth and ulji is preferably prepared by a manufacturing method comprising the following steps, but is not limited thereto.

(1) collecting and drying the hyacinths and scabs;

(2) extracting the extract by adding the extraction solvent after mixing the shredded hyacinth and the ulge;

(3) cooling the extract and then filtering;

(4) filtering the precipitate after sealing the filtered extract; And

(5) concentrating with a vacuum concentrator and drying.

Here, the curry and the large used in step (1) can be used without limitation, such as grown or commercially available.

The extraction of step (2) may be performed by shaking extraction, Soxhlet extraction, or a reflux extraction method, but is not limited thereto. It is preferable that extraction temperature is 50-120 degreeC, and it is more preferable that it is 80-100 degreeC. In addition, the extraction time is preferably 2 to 12 hours, the extraction frequency is preferably 1 to 5 times.

Filtration in the step (3) can be filtered to 100 ~ 500 mesh, it is preferred to use 300 mesh.

The filtered extract of step (4) may be subjected to a process of sinking the precipitate by leaving it at room temperature for 1 to 5 days, and the precipitate obtained through this is filtered twice with filter sheet of Edbentech No. 5 and Whatman GF / C 150 mm filter paper. It is preferable to make it.

Concentration of the step (5) may be made at 10 to 100 ℃ and preferably can be concentrated at 50 ℃ and preferably using a vacuum decompression concentrator or vacuum rotary evaporator. In addition, the drying is preferably using a vacuum drying, vacuum drying, boiling drying, spray drying or freeze drying method, more preferably a freeze drying method.

In another aspect, the present invention provides a cosmetic composition for improving atopic diseases, including a mixed extract of hyacinth and earlyge.

The mixed extract may be an extract extracted by mixing the hyacinth and the ratio in a weight ratio of 1: 0.1 to 1: 3, preferably a mixed extract extracted by mixing in a weight ratio of 1: 0.3 to 1: 3, more preferably It is a mixed extract extracted by mixing in a weight ratio of 1: 1 to 1: 3.

More preferably, the mixed extract is mixed with a weight ratio of 1: 1 to 1: 3 by using a 30 or 70% ethanol.

The atopy disease may be at least one selected from atopic dermatitis, allergic asthma, allergic rhinitis and allergic conjunctivitis, which is shown as an immune hypersensitivity reaction, and atopic dermatitis is more preferred.

In the present invention, the cosmetic composition is supple cosmetics, astringent cosmetics, nourishing cosmetics, nutrition cream, massage cream, essence, eye cream, eye essence, cleansing cream, cleansing foam, cleansing water, pack, powder, body lotion, body cream, It may be a formulation selected from the group consisting of body oils, body essences, makeup bases, foundations, hair dyes, shampoos, rinses and body cleansers.

The cosmetic composition of the present invention may be prepared in various forms according to a conventional cosmetic preparation method using the hydroxy and eroli mixed extract, stabilizers, solubilizers, vitamins, pigments, and the like commonly used in the cosmetic composition field Conventional adjuvants such as flavorings.

In addition, when the cosmetic composition is prepared in the form of cosmetic products, shampoos, hair lotions, hair creams, hair gels, etc. containing the mixed extracts of the eryri and ulge, it can be used diluted with conventional cleansing liquid, astringent liquid and moisturizing liquid have.

The cosmetic composition of the present invention is particularly preferably prepared in the form of lotion, lotion, cream, essence.

In the cosmetic composition of the present invention, the ulge and hyacinth mixed extract may be added to the cosmetic in an amount of 0.1% by weight to 10% by weight based on the total liquid weight of the cosmetic composition, and 0.001 to 5% by weight based on the total dry weight of the cosmetic composition. %, Preferably in an amount of 0.01 to 3% by weight.

In another aspect, the present invention provides a pharmaceutical composition for preventing or treating atopic dermatitis, including a mixed extract of hyacinth and ulji.

The atopy disease may be at least one selected from atopic dermatitis, allergic asthma, allergic rhinitis and allergic conjunctivitis, which is shown as an immune hypersensitivity reaction, and atopic dermatitis is more preferred.

The mixed extract of the hyacinth and ulji of the present invention is excellent in the effect of alleviating or improving the skin tickle, erythema, dryness in patients with atopic dermatitis can effectively improve the overall atopic dermatitis symptoms.

In the present invention, the pharmaceutical composition may be a formulation selected from the group consisting of ointments, gels, creams, patches and sprays. In addition, the pharmaceutical composition may be an oral or injectable preparation.

The oral preparations are powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, suspensions, solutions, emulsions or syrups, and the injectable preparations may be in the form of sterile injectables.

The pharmaceutical composition of the present invention may be formulated by adding a pharmaceutically acceptable carrier, and the formulation may be referred to Remington's Pharmaceutical Science (Recent Edition), Mack Publishing Company, Easton PA. The pharmaceutically acceptable carrier refers to those commonly used in preparing pharmaceutical compositions to those skilled in the art. For example, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pi Ralidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. Pharmaceutically acceptable carriers also include diluents or excipients, such as fillers, extenders, binders, wetting agents, disintegrants, surfactants and the like.

For example, pharmaceutical carriers and / or excipients suitable for topical administration are in accordance with the present invention conventional carriers and / or known to those skilled in the art in connection with pharmaceutical preparations for skin administration (eg for treating skin diseases). It is preferred that it is an excipient. Examples that may be mentioned are carriers suitable for producing dusting powders, emulsions, suspensions, oils, sprays, ointments, grease ointments, cream pastes, gels, foams, or solutions and suitable for transdermal therapeutic systems (TTS). And / or excipients. Topical pharmaceutical preparations of the invention may be semisolid formulations, in particular ointments (solution ointments, suspension ointments), creams, gels or pastes. Mainly used in the oil phase are fatty alcohols such as lauryl alcohol, cetyl alcohol, stearyl alcohol, fatty acids such as palmitic or stearic acid, liquid or solid paraffin or ozokerite, liquid to solid wax, for example For example isopropyl myristate, natural fats or some synthetic fats such as coconut fatty acid triglycerides, hardened oils such as hydrogenated peanuts or castor oils, or fatty acid partial esters of glycerol such as glycerol monostearate or glycerol Distearate. Suitable emulsifiers include surfactants such as fatty acid esters of nonionic surfactants such as polyalcohols or ethylene oxide adducts thereof such as polyglycerol fatty acid esters or polyoxy ethylene sorbitan fatty acid esters, sorbitan fatty acid esters such as Sorbitan oleate and / or sorbitan isostearate, etc., isostearates, sterols, or polyoxyethylene fatty alcohol ethers or fatty acid esters such as alkali metal salts of anionic surfactants such as fatty alcohol sulfonates, Examples are sodium lauryl sulfate, sodium cetyl sulfate or sodium stearyl sulfate and they are generally used in the presence of the fatty alcohols such as cetyl alcohol or stearyl alcohol. Among these, it is possible in particular to add an agent for preventing cream drying, for example, polyalcohols such as glycerol, sorbitol, propylene glycol and / or polyethylene glycol to the water phase, or preservatives, perfumes and the like to the water phase.

The pharmaceutical preparations of the present invention may be anhydrous ointment, contain paraffin, in particular low viscosity paraffin, which is suitable for topical use and liquid at body temperature, or the natural or partially synthetic fats such as coconut fatty acid trigly Cerides, hardened oils such as hydrogenated peanut or castor oils, fatty acid partial esters of glycerol such as glycerol monostearate and distearate, silicones such as polymethylsiloxanes such as hexamethyldisiloxane or octamethyltree It may contain siloxanes and may contain, for example, fatty alcohols associated with aqueous creams and increasing water absorption capacity, and sterols, wool waxes, other emulsifiers and / or other additives.

However, the present invention is not limited to the above-listed pharmaceutically acceptable carriers and the like, which are merely exemplary.

The applied amount of the hyacinth and allergy mixed extract contained in the pharmaceutical composition depends on the condition and weight of the patient, the extent of the disease, the drug form, the route of administration and the duration, but may be appropriately selected depending on the case. For example, the mixed extract of hyacinth and ulji can be applied to the skin at a dose of 0.0001 to 100 mg / kg, preferably 0.01 to 100 mg / kg, the application is divided once or several times a day You can also apply. In addition, the pharmaceutical composition may comprise 0.0001 to 50% by weight of the mixture of the hyacinth and scab mixed with respect to the total weight of the composition. The pharmaceutical composition can be applied to mammals such as humans by various routes, for example, by transdermal, oral, intravenous, intramuscular, or subcutaneous injection.

In addition, the pharmaceutical composition of the present invention may contain one or more active ingredients indicating improvement, alleviation, treatment or prevention of atopic diseases in addition to the mixed extract of hyacinth and ulji.

In addition, the pharmaceutical composition of the present invention can be used alone or in combination with methods using surgery, hormonal therapy, drug treatment and biological response modifiers for the improvement, alleviation, treatment or prevention of atopic diseases.

The numerical values set forth herein should be construed to cover equivalent ranges unless otherwise indicated.

Hereinafter, preferred examples, formulation examples, and experimental examples are presented to help understand the present invention. However, the following Examples, Formulation Examples and Experimental Examples are provided only to make the present invention easier to understand, and the content of the present invention is not limited by Examples, Formulation Examples or Experimental Examples.

Extraction solvent  star Heal  And early mix extract

Example  1-4.

Mixed extracts were prepared by using hyacinth and ulji collected from Wonju and nearby areas of Gangwon-do, Korea, and the extraction solvent was purchased from Daejeong Chemical. After collecting and drying, 5 kg of distilled water was added to the mixed sample of 250 g of chopped hyacinth and 750 g of dry matter (dry weight ratio = 1: 3), and the extract was extracted at 80 ° C. for 5 hours in an extractor equipped with a cooling condenser. Thereafter, the mixture was filtered through 300 mesh, the medicines and extracts were separated, sealed to prevent contamination, and left to stand at room temperature for 3 days, and then the precipitate was filtered twice with Edventech No. 5 filter paper and Whatman GF / C 150 mm filter paper. Then, the resultant was concentrated at 50 ° C. using a reduced pressure concentrator (BUCHI Co.) and dried using a lyophilizer (Ilshin Lab) to obtain a dry weight of 117.1 g (Example 1).

Except that the extraction solvent was used differently, the extracts of Examples 2 to 4 were obtained using the same method as above.

The mixed extracts of the prepared hyacinth and azule are shown in Table 1 below.

[Table 1]

mix By rate Heal  And early mix extract

Example  5 to 8.

After fixing the extraction solvent with 30% ethanol aqueous solution, the mixed extract was prepared by changing the mixing ratio of the ulge and hydroxy.

3 kg of 30% ethanol aqueous solution is added to a sample mixed with 500g of fine flakes and 500g of curry (dry weight ratio = 1: 1) after extraction and dried under a hot water condition at 80 ℃ for 5 hours in an extractor equipped with a cooling condenser. Filtered with 300 mesh to separate the medicine and extract. Sealed to prevent contamination and left at room temperature for 3 days, and the precipitate was filtered twice with Edbentech No. 5 filter paper and Whatman GF / C 150 mm filter paper. Then, the resultant was concentrated at 50 ° C. using a reduced pressure concentrator (BUCHI Co.) and dried using a lyophilizer (Ilshin Lab) to obtain a dry weight of 110.6 g (Example 5).

The mixed extract of Examples 6 to 8 was obtained by extracting in the same manner as above except for the mixing ratio of diary and sage.

The mixed extracts of the prepared hyacinth and azule are shown in Table 2 below.

[Table 2]

Formulation example  One. Heal  And using the mixed extract Cosmetic  Produce

1.1 Preparation of the lotion

1 kg of a lotion (skin lotion) containing an earl, hyacinth mixed extract powder (Example 2) was prepared. Raw materials and contents used are shown in Table 3 below.

[Table 3]

Specifically, glycerin, butylene glycol, propylene glycol, and preservatives were added to purified water in order and dissolved by stirring, and polyoxyethylene hardened castor oil was dissolved by heating to 60 ° C. Perfume was added to the melt to dissolve it, which was mixed with purified water again. Finally, the powder and ethanol of the mixture and extract of eryri and eryri of Example 2 of the present invention were added and stirred sufficiently to prepare a lotion.

1.2. Manufacture of lotions

A lotion containing a powder mixture, eryri extract (Example 2) 1kg was prepared in the content of Table 4 below.

[Table 4]

Specifically, the mixture of propylene glycol, carboxy polymer, preservative, and purified water in the content of Table 4 and heated to 80 ~ 85 ℃ while stirring the mixture, and then acting an emulsifier and beeswax, polysorbate 60, sorbitan sesquiol Elate, liquid paraffin, sorbitan stearate, lipophilic glycerin monostearate, stearic acid, glyceryl stearate / fig-400 stearate were heated to dissolve between 80 to 85 ° C. and then emulsified. After emulsification, triethanolamine was administered, neutralized with stirring using a stirrer, cooled to 50 ° C., then flavored, cooled to 35 ° C., and then mixed with Algear and hydroxy mixed extract powder (Example 2), cooled to 25 ° C. And aged to prepare a nutrient lotion.

1.3. Cream manufacturing

Earl, the cream containing the mixed extract extract (Example 2) 1kg to the content of Table 5 below was prepared.

[Table 5]

Specifically, the carboxyvinyl polymer, butylene glycol, triethanolamine, preservative, and purified water of the contents shown in Table 5 above and mixed stirring and heating to 80 ~ 85 ℃ to put into the manufacturing unit after the action of the emulsifier and stearic acid, cetyl Alcohol, glyceryl monostearate, polyoxyethylene sorbitan monostearate, sorbitan sesquioleate, glyceryl monostearate / glyceryl stearate / polyoxyethylene stearate, wax, liquid paraffin, squalane, caprylic Capric triglycerides were heated to 80-85 ° C. to dissolve and then emulsified. After emulsification, glycerin was added and neutralized with stirring using a stirrer. After cooling to 35 ° C., Algear and hydroxy mixed extract powder (Example 2) were added, cooled to 25 ° C., and aged to prepare nutrition cream. The nutrition cream prepared as described above was used as a prescription example in Experiment 6.

On the other hand, Comparative Example of Experimental Example 6 was prepared in the same manner as above except for the addition of Alge, eryri mixed extract powder of Example 2.

1.4. Essence manufacturing

Essence, containing an essence mixture extract powder (Example 2) 1kg was prepared in the content of Table 6 below.

[Table 6]

Specifically, non-ionic amphiphilic lipids were prepared by homogenizing cytosterol, polyglyceryl 2-oleate, glyceramide, steares-4, and cholesterol in a content as shown in Table 6 at a constant temperature. Mix this nonionic amphiphilic lipid with Alge, hyacinth mixed extract powder (Example 2), dicetyl phosphate, macadamia oil and purified water, and homogenize at a constant temperature to pass through the microfluidizer, followed by 50 ~ 60 Warmed to 占 폚 and slowly added to homogenization and again passed through a microfluidizer. Thereafter, carboxyvinyl polymer, xanthan gum, preservative, and purified water were added and dispersed to stabilize and mature to prepare an essence.

Formulation example  2. Heal  And preparation of medicines using the mixed extract

2.1 Preparation of Tablets

The tablets containing the mixture, eryri mixed extract powder (Example 2) was prepared in the content of Table 7.

[Table 7]

After mixing the above components, tablets were prepared by tableting according to the usual preparation method of tablets.

2.2 Preparation of Injection

Injectables containing a mixture of erythritis, hyacinth extract extract (Example 2) were prepared in the following Table 8.

[Table 8]

(2 ml) per 1 ampoule in accordance with the usual injection method.

2.3 Preparation of Liquids

Earl, hydroxy mixed extract powder (Example 2) containing a liquid formulation was prepared in the following Table 9.

TABLE 9

Purified water was added and adjusted to a total volume of 1,00 ml, and the above ingredients were mixed according to a conventional method for preparing a liquid, and then filled into a brown bottle and sterilized to prepare a liquid.

Experimental Example  1. Confirm cytotoxicity and cell proliferation effect

In order to confirm the cytotoxicity of the mixed extract of Examples 1 to 8, the cell proliferation effect of the mixed extract dry powder was measured. Cells (fibroblasts, 3T3) were dispensed in a 96 well microplate at 5,000 cells / well and incubated in a thermostat for 30 minutes, and Examples 1-8 samples were 0.01, 0.05, 0.1, 0.5, 1.0 (%, W / V, respectively). ) And incubated for 72 hours. Thiazoline blue was then administered and further incubated for 4 hours. The culture medium was discarded, and acidic isopropanol was added to each well of the microplate. After stirring for 5 minutes, absorbance was measured at 570 nm. 10% Fetal Bovine Serum (FBS) medium was used as the amount of sample injected and co-cultured at the optimum conditions for cell growth. As a control, the cell proliferation rate of the control group was 100% and the cell proliferation rate was calculated. .

Cell proliferation was calculated using the following equation 1.

[Formula 1]

Cell growth rate (%) of Examples 1 to 8 calculated by Equation 1 is shown in Table 10.

[Table 10]

As shown in Table 10, when the cell proliferation at the optimal conditions for cell growth was 100%, Examples 1 to 8, which are mixed extracts of Algebra and hyacinth, exhibited no less than 100% cell proliferation rate and were not toxic to cells. It was confirmed that there is a cell proliferation effect.

Experimental Example  2. Free radical  Confirm elimination effect

Since the extract dried powders prepared in Examples 1 to 8 were all non-cytotoxic and excellent in cell proliferation effect, the free radical scavenging effect was obtained by using Example 2 having the highest cell proliferation effect among them. The measurement confirmed the antioxidant effect. As a comparative substance, quercetin, a representative antioxidant, was used. Experiments were performed using the DPPH (1,1-diphenyl-2picryl-hydrazyl) method (Blois. MS Nature 181, 1190, 1958), and DPPH and quercetin were used by SIGMA. . 2 ml of various concentrations (5, 10, 20, 30 ppm) of Example 2 were added to 1 ml of 0.2 mM DPPH methanol solution, and an ethanol solution of quercetin was added as a control, which was stirred well and reacted at room temperature for 10 minutes. After the reaction, the absorbance was measured at 517 nm, and purified water was used instead of each sample in the blank test.

Free radical scavenging effect was calculated using Equation 2 below.

[Formula 2]

Table 11 shows the free radical scavenging effect (%) of Example 2 calculated by Equation 2.

[Table 11]

As shown in Table 11, the free radical 50% scavenging concentration (SC ₅₀ ) of Example 2, which is a mixture of hydroxy and allergy, was 13.53 ppm, which showed a similar level of free radical scavenging effect to quercetin, a representative antioxidant. Since quercetin is a single substance, considering the process cost and stability of use to obtain pure substance by extracting it, the natural plant extracts of hydroxya and alsier mixture extract are superior in terms of effectiveness and cost as antioxidants.

Experimental Example  3. Lipoxygenase  Check the inhibitory effect

The inhibitory effect of lipoxygenase activity was measured on the extract powders prepared in Examples 1 to 3 and 8, and quercetin was used as a comparative substance. The experiment was performed using the TBARS method, and linoleic acid, lipoxygenase, thiobarbitulic acid, and quercetin used in the experiment were used by SIGMA. . After adding 0.05 ml of Examples 1 to 3, Example 8 and quercetin at various concentrations (1, 5, 10, 20, 30, 50 ppm) to 1 ml of linoleic acid at 1 mM, 0.95 ml of lipoxygenase was administered, It stirred and reacted for 10 minutes at 25 degreeC. 0.5 ml of trichloroacetic acid was added, 1 ml of thiobarbiturlic acid was added, and then heated for 10 minutes. The reaction was terminated by cooling in ice water for 2-3 minutes. 2 ml of butanol was added to the reaction solution, followed by centrifugation at 4000 g for 5 minutes, and the absorbance was measured at 535 nm. At this time, purified water was used instead of each sample as a control.

Equation 3 below was used to calculate the inhibitory effect of lipoxygenase activity.

[Formula 3]

Table 12 shows the inhibitory effect (%) of lipoxygenase activity of Examples 1 to 3 and Example 8 calculated by Equation 3.

[Table 12]

As shown in Table 12, the lipoxygenase 50% activity inhibitory concentration (IC ₅₀ ) of Example 2 was 15.55 ppm, which was superior to that of 16.46 ppm of quercetin, and Example 3 showed a lipoxygenase activity similar to that of quercetin. It was confirmed to exhibit an inhibitory effect. However, Example 1, which is a water extract among the mixed extracts of hyacinth and early larvae, was found to have a lower effect of inhibiting lipoxygenase activity than Examples 2, 3, and 8, which are ethanol extracts, and the hyacinth and alsies were mixed at a ratio of 3: 1. Example 8 also showed a lower effect of inhibiting lipoxygenase activity than Examples 2 and 3, which had a higher ratio of allergy in the mixture. It is known that the anti-inflammatory effect is excellent when lipoxygenase activity is inhibited. Therefore, according to the results, the mixed extract of hyacinth and sage may have an anti-inflammatory effect. In particular, the ratio of sage is high and ethanol extract is the highest. It was confirmed that the anti-inflammatory effect.

Experimental Example  4. Hyaluronidase  Identification of the activity inhibition effect

The inhibitory effect of hyaluronidase activity of the extract dry powder of Examples 1 to 8 was measured. By applying the Morgan-Elson method, the final enzyme activity of hyaluronidase is 400NF unit / ml, the final concentration of hyaluronic acid is 0.4mg / ml, and the active compound Compound 48/80 buffer (buffer) solution) (0.1 mg / ml) was used to measure the activity of hyaluronidase. A green tea extract known to have hyaluronidase inhibitory activity was used as a sample solution by dissolving the sample in a buffer solution. In each experiment, a buffer solution was used instead of an enzyme solution.

The following formula 4 was used to calculate the hyaluronidase activity inhibitory effect.

[Formula 4]

Table 13 shows the inhibitory effect (%) of hyaluronidase activity of Examples 1 to 8 calculated by Equation 4.

[Table 13]

As can be seen in Table 13, the hyaluronidase 50% activity inhibitory concentration (IC ₅₀ ) of Examples 1 to 8 of the present invention was 1.06% to 2.75%, which was very excellent in inhibiting hyaluronidase activity. These values are similar to or lower than 2.47% of the control group green tea extract, so it can be confirmed that the hyaluronidase activity inhibitory effect of the present invention is very excellent. In particular, Examples 2, 3, 4, and 7, which have a high mixing ratio in ethanol extracts, were found to have a relatively better effect of inhibiting hyaluronidase activity compared to Examples 6 and 8, which had a high mixing ratio of hydroxy. .

Experimental Example  5. Immune Cells Degranulation  Inhibitory effect

Degranulation effect of immune cells was measured using the extract dry powder of Examples 1 to 8. Degranulation of immune cells is known to be a cause of skin inflammatory response, and by the degranulation of mast cells, various inflammation mediated immune modulators are excessively secreted. Therefore, if the degranulation of immune cells can be effectively suppressed, the inflammatory response of the skin can be suppressed. The extracts of Examples 1 to 8 were treated with RBL-2H3 cell line (Rat mast cell line) to confirm the effect of inhibiting degranulation by antigen. RBL-2H3 cell line was obtained from Eagle's medium (Dulbeccos' modified Eagle's medium, DMEM) containing 1% Fetal bovine serum and L-glutamine. Were cultured in a 37 ° C., 5% CO ₂ incubator, and the cells with adherence were suspended using trypsin-EDTA solution, separated, recovered and used for experiments. After activating the RBL-2H3 cell line with an activator (Dinitrophenol-conjugated human serum albumin, (DNP-HSA) -specific IgE), the activated cells were treated with 50 ng / mL antigen (DNP-HSA) and Example 1 To 8 were incubated for 60 minutes with 0, 5, 25 and 125 μg / ml respectively. Then, the amount of β-hexosaminidase secreted into the medium was measured using an ELISA reader. Inhibition of β-hexosaminidase secretion means preventing degranulation of immune cells.

The results are shown in Table 14.

[Table 14]

As shown in Table 14, the degranulation 50% inhibitory concentration (IC ₅₀ ) of the immune cells of Examples 1 to 8 was 22.49 to 53.41 ug / ml, which can be found to be very effective for degranulation of immune cells. Examples 2 to 4 and 7 having a high mixing ratio of ethanol in the ethanol extract showed a more effective suppression of immune cell degranulation compared to Examples 6 and 8 having a high mixing ratio of hydroxy, thus inhibiting the most effective skin inflammatory response. The effect was expected.

Experimental Example  6. Atopic skin improvement effect confirmed

The mixed extracts of the hyacinth and ulji of Example 2 were prepared in a cream formulation according to Formulation Example 1.3 above, and the clinical evaluation of the atopic skin improvement effect was measured. In order to evaluate the improvement effect of atopic skin, the following experiments were conducted on 30 children who were treated with atopic skin and visited to Korean medicine and hospital. The test sample was applied to the whole body by dividing the right side and the left side by the double-blinded test, but the use of other moisturizers that could affect the effect of the test sample was prohibited. 1, 2, 4, 8 weeks after application of the test sample was evaluated by SCORAD SCORAD (SCORing Atopic Dermatitis) measurement to confirm the improvement effect of atopic skin. I made it.

The improvement effect of atopy skin was judged using accuracy criteria, intensity criteria, and subjective criteria.

-Criteria

① Extent criteria: Area = damage area / 100

② intensity criteria

-Erythema (1/2/3)

-Edema (1/2/3)

-Exudate (1/2/3)

-Skin peeling (1/2/3)

Taekwondo degree (1/2/3)

-Drying degree (1/2/3)

Subjective criteria

-Itching (1-10)

Insomnia (1-10)

* Calculation of coderad = (degree of criterion / 5) + (strength criterion / 7) + subjective criterion

Atopic dermatitis improvement effect measurement results using the above criteria are shown in Table 15, and the skin of atopic dermatitis patients before and after applying the cream containing the mixed extract of Example 2 is shown in FIG.

[Table 15]

As shown in Table 15, the cream of Example 2 containing the mixed extract of hyacinth and ulzzi showed a very good atopic skin improvement effect compared to the comparative formulation containing simply water. In particular, after 8 weeks it was confirmed that the excellent improvement effect of more than 90%. In addition, as shown in Figure 1 after applying the cream containing the mixed extract of Example 2 on the skin of atopic skin patients, the improvement effect of the dermatitis was clearly confirmed with the naked eye.

On the other hand, in order to examine the atopic dermatitis mitigation effect felt by the caregiver, the cream of Example 2 is to be used for one month, and the parent of the user is satisfied with itching, erythema change, dry skin of the skin according to the criteria shown in Table 16 below. In addition, the overall degree of dermatitis improvement and side effects of test samples were evaluated. As a comparative example, a cosmetic containing water was used for one month.

[Table 16] Judging Criteria

[Table 17]

As shown in Table 17, the cream cosmetic of Example 2 containing a mixed extract of hyacinth and ulge has been shown to improve skin itch, erythema change, dry skin, etc. in children with atopic skin as a subject. It was evaluated to have an effect of improving dermatitis. In addition, the satisfaction of the side effects were also high, and it was confirmed that the mixed extracts of hydroxy and uresis, which are natural substances, could improve the atopic dermatitis as a whole without side effects.

Claims (8)

 Antioxidant and anti-inflammatory cosmetic composition comprising an alcohol extract of hyacinth and earlyge mixed in a weight ratio of 1: 1 to 1: 3. delete The cosmetic composition according to claim 1, wherein the alcohol is a lower alcohol having 1 to 4 carbon atoms. The cosmetic composition of claim 3 wherein the alcohol is 30% (v / v%) or 70% (v / v%) ethanol. delete delete  Cosmetic composition for improving atopic diseases comprising alcohol extract of hyacinth and earlyge mixed in a weight ratio of 1: 1 to 1: 3.  A pharmaceutical composition for preventing or treating atopic diseases, including alcohol extracts of hyacinth and ulji mixed in a weight ratio of 1: 1 to 1: 3.

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