KR20200101113A - A composition for improving, preventing and treating dermatitis comprising Chrysanthemum zawadskii var extract - Google Patents

Abstract

The present invention relates to a composition for preventing, alleviating and treating inflammation. By containing a Siberian chrysanthemum flower extract or a Siberian chrysanthemum stem extract as an active ingredient, diseases that may be caused by inflammation can be alleviated, prevented or treated. In addition, since the present invention is not toxic, the present invention can be consumed in the form of food.

Description

A composition for improving, preventing and treating dermatitis comprising Chrysanthemum zawadskii var extract} containing Gujeolcho extract as an active ingredient

The present invention relates to a composition containing as an active ingredient Gujeolcho extract capable of preventing or treating inflammation.

The inflammatory response is one of the defense reactions of living tissues against external stimuli. It is involved in the physiological and pathological processes of various diseases, and is a mechanism to repair or regenerate tissues damaged by external stimuli. The characteristic of this inflammatory reaction is that it involves physicochemical, chemical, and bacteriological changes in many other tissues, such as skin tissue and joint tissue.

That is, the inflammatory response is activated by the immune cells recognizing foreign substances in a living body, and the activated immune cells secrete many factors that mediate inflammation. These inflammatory phenomena cause a large increase in various types of polykaryotic leukocytes (PMNs) and immune substances, and these increased cells secrete various types of proteolytic enzymes and cytokines, which are inflammatory cell products, to provide treatment and defense. Allows you to

Symptoms caused by such inflammation include fever, redness, and swelling, and it has been reported that various incurable diseases such as dementia, cardiovascular disease, cancer, obesity, arthritis, diabetes, etc. can occur due to continuous inflammatory reaction. When an inflammatory reaction occurs, inflammatory mediators such as nitric oxide (NO), prostaglandin E2 (PGE2), and inflammatory cytokines are secreted, and nitric oxide (NO) is an indicator of the inflammatory reaction, and nitric oxide synthase from L-arginine ( NOS).

The formation of nitric oxide (NO) also plays a role in the immune response, but nitric oxide (NO) excessively produced by iNOS, which is expressed by lipopolysaccharide (LPS) or inflammatory cytokines, is interleukin (IL)-1β. , IL-6, IL-8, and tumor necrosis factor (TNF)-α by generating pro-inflammatory cytokines, which intensifies the inflammatory response, causing tissue damage, genetic mutation, and nerve damage.

Existing anti-inflammatory agents are largely classified as steroidal and nonsteroidal anti-inflammatory agents. Most synthetic anti-inflammatory agents have various side effects in addition to their main action, so there is an urgent need to develop anti-inflammatory drugs with excellent effects and less side effects. In particular, the initial response to inflammation through inhibition of nitric oxide (NO) generation has become a major target of anti-inflammatory drug development.

Therefore, natural extracts are required that have excellent anti-inflammatory effects by inhibiting the production of NO, which is an inflammatory mediator, through natural materials, have excellent antioxidant effects, and have no side effects.

Korean Patent Registration No. 1906856 Republic of Korea Patent Publication No. 2017-0134237

An object of the present invention is to provide a pharmaceutical composition containing a Gujeolcho flower extract or Gujeolcho stem extract capable of preventing or treating inflammation as an active ingredient.

In addition, another object of the present invention is to provide a health functional food containing a Gujeolcho flower extract or Gujeolcho stem extract capable of preventing or improving inflammation as an active ingredient.

In addition, another object of the present invention is to provide a cosmetic composition containing a Gujeolcho flower extract or Gujeolcho stem extract as an active ingredient to have anti-inflammatory or antioxidant activity.

The pharmaceutical composition capable of preventing or treating inflammation of the present invention for achieving the above object may contain Gujeolcho flower extract or Gujeolcho stem extract as an active ingredient.

The Gujeolcho stem extract may be an extract of a mixture of Gujeolcho stem and leaves in a weight ratio of 1:0.1-0.8.

In addition, the health functional food capable of preventing or improving the inflammation of the present invention for achieving the other object described above may contain a Gujeolcho flower extract or Gujeolcho stem extract as an active ingredient.

In addition, the cosmetic composition having anti-inflammatory or antioxidant activity of the present invention for achieving the another object described above may contain a Gujeolcho flower extract or a Gujeolcho stem extract as an active ingredient.

The composition containing the Gujeolcho flower extract or Gujeolcho stem extract of the present invention as an active ingredient has excellent antioxidant or anti-inflammatory activity. Therefore, the Gujeolcho flower extract or Gujeolcho stem extract of the present invention is a material having various functions and can be usefully used in various industrial fields such as pharmaceuticals, foods, and cosmetics.

In particular, Gujeolcho is a natural medicinal plant and can be edible, so the composition of the present invention comprising an extract derived therefrom as an active ingredient has a safe advantage even for long-term use.

The present invention relates to a composition containing a Gujeolcho extract as an active ingredient so as to prevent, improve or treat inflammation.

The composition of the present invention can be used for various purposes and uses requiring antioxidant or anti-inflammatory activity, and specifically, a functional material capable of imparting antioxidant activity to articles applied in various industrial fields such as pharmaceuticals, cosmetics, food and animal feed. It can be used as a pharmaceutical preservative, cosmetic preservative, food preservative, pharmaceutical additive, cosmetic additive, food additive and feed additive.

Hereinafter, the present invention will be described in detail.

The composition having the antioxidant activity of the present invention and capable of preventing, improving or treating inflammation contains Gujeolcho extract, preferably Gujeolcho flower extract or Gujeolcho stem extract as an active ingredient.

Gujeolcho ( Chrysanthemum zawadskii var.latilobum (Maxim.) Kitam.) used in the present invention is a perennial herb of the chrysanthemum family, and is also referred to as yaguk (野菊), sunmocho (仙母草), gobong (苦蓬), wild chrysanthemum. It grows wildly in mountainous and highlands nationwide, and grows in China, Russia, Mongolia, and Japan, and has been used in the form of a pill or decoction in the form of a pill after collecting stems and leaves in the private sector in the fall. It reproduces while the rootstock in the ground extends sideways, and the flowering period is August-October and the fruiting period is October-November. The herbal names of Gujeolcho outpost are called Seonmocho, Gujeolcho, and the flowers are elegant and beautiful, so they are widely cultivated for landscape creation. Gujeolcho outpost and flower spikes have been used for pneumonia, bronchitis, cough, cold, women's disease, poor circulation, gastrointestinal disease and high blood pressure.

Gujeolcho extract according to the present invention is a Gujeolcho flower extract or Gujeolcho stem extract, wherein the Gujeolcho stem extract is an extract of a mixture of Gujeolcho stems and leaves in a weight ratio of 1:0.1-0.8, preferably 1:0.2-0.5 to be. When the amount of leaves based on the stem is less than the lower limit, the antioxidant and anti-inflammatory effects may be poor, and when the content of the leaf is higher than the upper limit, it is not preferable for preventing, treating or improving inflammation.

The extract of the present invention is an extract extracted for 20 to 30 hours, preferably 24 to 28 hours at 80 to 100 ℃. When the extraction temperature and time are less than the lower limit, the active ingredient may not be extracted, and when the extraction temperature and time are greater than the upper limit, the burnt taste is strong and the nutrient component may be destroyed.

The extraction solvent is not particularly limited, but may include water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof, and the extract extracted with water has superior antioxidant and anti-inflammatory effects compared to the extract extracted with an organic solvent. Therefore, it is preferable to use water as an extraction solvent.

The Gujeolcho flower extract or Gujeolcho stem extract prepared as described above may be used as an extract itself or may be powdered for convenience. In the method of powdering the Gujeolcho flower extract or Gujeolcho stem extract, the extract is concentrated under reduced pressure to reduce the volume, and then freeze-dried with a freeze dryer to powder.

Gujeolcho flower extract or Gujeolcho stem extract of the present invention has a meaning in a broad sense to include Gujeolcho flower or Gujeolcho stem processed product, such as Gujeolcho flower or Gujeolcho stem powder, formulated to administer Gujeolcho flower or Gujeolcho stem to an animal. . Although the experiment was conducted with Gujeolcho flower extract or Gujeolcho stem extract in the present invention, it will be predictable to those skilled in the art that the desired effect can be achieved even in the form of Gujeolcho flower or Gujeolcho stem processed product.

Meanwhile, in the present specification, the term “containing as an active ingredient” means including an amount sufficient to achieve the efficacy or activity of the Gujeolcho flower extract or Gujeolcho stem extract. As an example, the mixture containing the Gujeolcho flower or Gujeolcho stem is used in a concentration of 10 to 1500 µg/ml, preferably 100 to 1000 µg/ml. Since the mixture containing Gujeolcho flowers or Gujeolcho stems is a natural product and does not have side effects on the human body even if excessively administered, the quantitative upper limit of the mixture containing Gujeolcho flowers or Gujeolcho stems included in the composition of the present invention is selected within an appropriate range by those skilled in the art. can do.

The pharmaceutical composition of the present invention may be prepared using a pharmaceutically suitable and physiologically acceptable adjuvant in addition to the active ingredient, and the adjuvant includes excipients, disintegrants, sweeteners, binders, coating agents, expanding agents, lubricants, It is possible to use a lubricant or flavoring agent.

For administration, the pharmaceutical composition may contain one or more pharmaceutically acceptable carriers in addition to the above-described active ingredients, and may be preferably formulated into a pharmaceutical composition.

The formulation form of the pharmaceutical composition may be granules, powders, tablets, coated tablets, capsules, suppositories, solutions, syrups, juices, suspensions, emulsions, drops, or injectable solutions. For example, for formulation in the form of tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like. In addition, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included in the mixture. Suitable binders are, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, lacquercanth or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like.

As acceptable pharmaceutical carriers for compositions formulated as liquid solutions, sterilization and biocompatible, saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and One or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added as necessary. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to prepare injectable formulations such as aqueous solutions, suspensions, emulsions, etc., pills, capsules, granules, or tablets.

Furthermore, it can be preferably formulated according to each disease or ingredient using a method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA as an appropriate method in the field.

The pharmaceutical composition of the present invention may be administered orally or parenterally, and in the case of parenteral administration, it may be administered by intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, etc., preferably oral administration. .

A suitable dosage of the pharmaceutical composition of the present invention varies depending on factors such as formulation method, mode of administration, age, weight, sex, pathological condition, food, administration time, route of administration, excretion rate and response sensitivity of the patient, Usually, the skilled practitioner can readily determine and prescribe the dosage effective for the desired treatment or prophylaxis. According to a preferred embodiment of the present invention, the daily dosage of the pharmaceutical composition of the present invention is 0.001-10 g/kg.

The pharmaceutical composition of the present invention is prepared in unit dosage form by formulating using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily carried out by a person having ordinary knowledge in the art. Or it can be made by incorporating it into a multi-dose container. At this time, the formulation may be in the form of a solution, suspension, or emulsion in an oil or aqueous medium, or may be in the form of an extract, powder, granule, tablet or capsule, and may additionally include a dispersant or a stabilizer.

In addition, the present invention provides a food composition that has an antioxidant activity containing Gujeolcho flower extract or Gujeolcho stem extract as an active ingredient, and can prevent, treat or improve inflammation.

The food composition according to the present invention may be formulated in the same manner as the pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, alcoholic beverages, confectionery, diet bars, dairy products, meat, chocolate, pizza, ramen, other noodles, gum, ice cream, vitamin complexes, health supplements. Etc.

The food composition of the present invention may include not only a mixture containing Gujeolcho flower extract or Gujeolcho stem extract as an active ingredient, but also ingredients commonly added during food production, for example, proteins, carbohydrates, fats, nutrients, It includes seasoning and flavoring agents. Examples of the aforementioned carbohydrates include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides, and the like; And polysaccharides, for example, common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents, natural flavoring agents [taumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.]) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is made of drinks and beverages, in addition to the mixture containing the Gujeolcho flower extract or Gujeolcho stem extract of the present invention, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, and various plant extracts And the like may additionally be included.

The present invention provides a health functional food comprising a food composition for preventing, treating, or improving inflammation comprising the Gujeolcho flower extract or Gujeolcho stem extract as an active ingredient. Health functional foods are foods prepared by adding Gujeolcho flower extract or Gujeolcho stem extract to food materials such as beverages, teas, spices, chewing gum, confectionery, etc., or prepared by encapsulation, powdering, suspension, etc. In this case, it means that it has a specific effect on health, but unlike general drugs, it has the advantage of not having side effects that may occur when taking the drug for a long time by using food as a raw material. The health functional food of the present invention obtained in this way is very useful because it can be consumed on a daily basis. The amount of Gujeolcho flower extract or Gujeolcho stem extract in such health functional foods cannot be uniformly regulated depending on the type of health functional food to be targeted, but can be added within the range that does not damage the original taste of the food. It is usually in the range of 0.01 to 50% by weight, preferably 0.1 to 20% by weight, based on the food. In addition, in the case of health functional foods in the form of pills, granules, tablets or capsules, it is usually added in the range of 0.1 to 100% by weight, preferably 0.5 to 80% by weight. In one embodiment, the health functional food of the present invention may be in the form of a pill, tablet, capsule or beverage.

In addition, the present invention provides a use of a Gujeolcho flower extract or Gujeolcho stem extract for the manufacture of a pharmaceutical or food for preventing, treating or improving inflammation. As described above, Gujeolcho flower extract or Gujeolcho stem extract may be used for inflammation.

In addition, the present invention provides a method for improving, preventing or treating inflammation comprising administering an effective amount of Gujeolcho flower extract or Gujeolcho stem extract to a mammal.

The term "mammal" as used herein refers to a mammal that is the subject of treatment, observation or experimentation, and preferably refers to a human.

The term "effective amount" as used herein refers to the amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human, which is considered by a researcher, veterinarian, doctor or other clinician, Includes an amount that induces relief of symptoms of the disease or disorder. It is apparent to those skilled in the art that the effective amount and the number of administrations for the active ingredient of the present invention will vary depending on the desired effect. Therefore, the optimal dosage to be administered can be easily determined by those skilled in the art, and the type of disease, the severity of the disease, the content of active ingredients and other ingredients contained in the composition, the type of formulation, and the age, weight, and general health of the patient. It can be adjusted according to various factors including condition, sex and diet, time of administration, route of administration and secretion rate of the composition, duration of treatment, and drugs used simultaneously. In the prevention, treatment or improvement method of the present invention, in the case of adults, it is preferable to administer thistle and onion at a dose of 0.001 g/kg to 10 g/kg when administered once to several times a day.

The composition comprising Gujeolcho flower extract or Gujeolcho stem extract as an active ingredient in the treatment method of the present invention can be used for oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal routes. It can be administered in a conventional manner.

In addition, in the cosmetic composition of the present invention, in addition to the cosmetic composition described above, other ingredients that are usually blended in the cosmetic may be blended as needed, and such blending ingredients include water, oil, surfactants, moisturizers, thickeners, fragrances, preservatives, At least one selected from the group consisting of a neutralizing agent, an emollient agent, a skin protectant, and a skin conditioning agent may be mentioned.

In addition, the cosmetic composition may be prepared in the form of an emulsifying formulation and a solubilizing formulation commonly used in the art, for example, skin, emulsion, essence, lotion, essence, body lotion, body gel, body essence, body cleanser, cleansing Foam, cleansing cream, cleansing gel, pack, massage cream, massage gel, nutritional cream, sleep cream, and moisture cream may include any one formulation selected from the group consisting of.

Hereinafter, a preferred embodiment is presented to aid the understanding of the present invention, but the following examples are only illustrative of the present invention, and it is obvious to those skilled in the art that various changes and modifications are possible within the scope and spirit of the present invention, It is natural that such modifications and modifications fall within the appended claims.

Example 1. Gujeolcho flower extract

Hot air dried Gujeolcho flower 50 g and purified water 50 ml were mixed, extracted with hot water at 100° C. for 24 hours, filtered under reduced pressure, concentrated and freeze-dried to obtain Gujeolcho flower extract powder.

Example 2. Gujeolcho stem extract

However, in the same manner as in Example 1, Gujeolcho stem and leaves were mixed in a weight ratio of 1:0.4 instead of Gujeolcho flowers to obtain Gujeolcho stem extract powder.

Comparative Example 1. Gujeolcho leaf extract

It was carried out in the same manner as in Example 1, but a Gujeolcho leaf extract powder was obtained using Gujeolcho leaves instead of Gujeolcho flowers.

Comparative Example 2. Gujeolcho stem extract

It was carried out in the same manner as in Example 1, but a Gujeolcho stem extract powder was obtained by using a Gujeolcho stem instead of a Gujeolcho flower.

<Test Example>

Test Example 1. Cell stability measurement

To measure the toxicity to RAW264.7 cells of Gujeolcho extract prepared in Examples and Comparative Examples. Cytotoxicity was compared by measuring the survival rate using MTT{3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide}assay. Macrophage RAW264.7 was cultured in DMEM medium supplemented with 10% fetal bovine serum at 37°C. The cultured cells were cultured in a 96-well plate at a concentration of ⁵ ×10 ⁴ cells/well for 24 hours. The samples are diluted in DMEM medium at concentrations of 100, 200, 400, 800, and 1000 μg/mL, respectively. Remove the medium from the cultured plate, dispense the diluted sample into the medium and incubate for 24 hours, and react by adding 10 μL of 5 mg/mL MTT to each well. After 3 hours, the culture solution was removed, and 100 μL of dimethyl sulfoxide (DMSO) was dispensed into each well, reacted for 15 minutes, and absorbance was measured at 595 nm. The control group was compared with the sample treatment group as when the sample diluted in the medium was dispensed, and only the medium was dispensed. Cell viability (%) was calculated according to the following [Equation 1]. At this time, the cell viability of the control group was 100%.

[Equation 1]

Cell viability (%) = (A/B)X100

A: absorbance of hot water extract treatment group

B: absorbance of the control group

division Concentration (㎍/㎖) 100 200 400 800 1000 Example 1 98.2 98.2 98.0 97.9 97.1 Example 2 99.2 99.0 99.0 98.2 97.0 Comparative Example 1 98.1 98.1 97.5 97.3 96.8 Comparative Example 2 98.0 97.5 97.4 97.1 96.7

At this time, the cell viability of the control group was 100%.

As shown in Table 1 above, the Gujeolcho flower extract prepared according to Example 1 of the present invention and the Gujeolcho stem extract prepared according to Example 2, as well as the extracts of Comparative Examples 1 and 2 were also up to a concentration of 1000 μg/mL. It was confirmed that no cytotoxicity appeared.

Test Example 2. Measurement of antioxidant

2-1. Antioxidant activity (mgVCEA/g _sample ): Using ABTS (2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid)) to measure the antioxidant effect of Gujeolcho extract prepared in Examples and Comparative Examples . The blue ABTS radical absorbs light with a wavelength of 734 nm, and antioxidant activity can be measured by measuring the absorbance that changes as it is decolorized by an antioxidant. Measurement method is ABTS radical solution 1.0 mM AAPH (2,2 ??azobis-(2-amidinopropane)HCl, Sigma A440914) and 2.5 mM ABTS2-(2,2 -azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) ) diammonium salt, Sigma A1888) was dissolved in 100 mL of PBS [(40mg AAPH + 205mg ABTS2-)/100 mL of PBS], heated for 30 minutes, cooled at room temperature, and then filtered and diluted to obtain an absorbance value of 0.7 Dilute to about enough to use. After diluting each sample to a concentration of 5 mg/mL, 30 μL was subjected to dark reaction with 1470 μL of ABTS radical solution at 37°C for 10 minutes. After the reaction, the absorbance at 734 nm was measured, and the measured absorbance was converted from the standard curve using vitamin C and expressed as VCEAC (Vitamin C Equivalent Antioxidant Capacity).

2-2. Total polyphenol content (mgGAE/g _sample ): The total polyphenol content of the Gujeolcho extract prepared in Examples and Comparative Examples was measured using the Folin-Denis method. After diluting each sample to a concentration of 5 mg/mL, 0.5 mL was reacted with 1 mL of 2% Na ₂ CO _{3 for} 3 minutes, 50 μL of Folin-Ciocalteu's phenol reagent was added, reacted for 3 minutes, and the absorbance was measured at 720 nm. I did. The standard curve was prepared using gallic acid, and the total polyphenol content of the extract was converted.

2-3. Total flavonoid content (mgRE/g _sample ): The total flavonoid content of the Gujeolcho extract prepared in Examples and Comparative Examples was measured. After each sample was diluted to a concentration of 5 mg/mL, 1 mL was reacted with 75 μL of 5% NaNO ₂ for 5 minutes, and then 150 μL of 10% AlCl _{3 was} added to react for 6 minutes. After reacting with 500 μL of 1 M NaOH for 11 minutes, absorbance was measured at 510 nm. The standard curve was prepared using rutin, and the total flavonoid content of the extract was converted.

division Antioxidant activity
(mgVCEAC/g) Total polyphenol content (mgGAE/g) Total flavonoid content (mgRE/g) Example 1 34.2 41.3 27.4 Example 2 48.6 53.5 45.9 Comparative Example 1 11.3 20.1 11.3 Comparative Example 2 9.5 13.8 5.8

As shown in Table 2 above, it was confirmed that the antioxidant activity of 1 g of Gujeolcho flower extract prepared according to Example 1 of the present invention was equivalent to the antioxidant activity of 34.2 mg of vitamin C, and Gujeolcho stem extract prepared according to Example 2 It was confirmed that the antioxidant activity of 1 g was equivalent to that of 48.6 mg of vitamin C. On the other hand, the antioxidant activity of each 1 g of the Gujeolcho leaf extract of Comparative Example 1 and the Gujeolcho stem extract of Comparative Example 2 was equivalent to the antioxidant activity of 11.3 mg and 9.5 mg of each vitamin C, and the antioxidant activity was lower than that of Examples 1 and 2. Confirmed.

In addition, it was confirmed that the total polyphenol content of 1 g of Gujeolcho flower extract prepared according to Example 1 of the present invention was equal to 41.3 mg of gallic acid, and the total polyphenol content of 1 g of Gujeolcho stem extract prepared according to Example 2 was It was confirmed to be equivalent to 53.5 mg of gallic acid. On the other hand, the total polyphenol content of each 1 g of the Gujeolcho leaf extract of Comparative Example 1 and the Gujeolcho stem extract of Comparative Example 2 was equivalent to 20.1 mg and 13.8 mg of gallic acid, respectively, and the total polyphenol content was lower than that of Examples 1 and 2. Confirmed.

In addition, it was confirmed that the total flavonoid content of 1 g of Gujeolcho flower extract prepared according to Example 1 of the present invention was equal to 27.4 mg of rutin, and the total flavonoid content of 1 g of Gujeolcho stem extract prepared according to Example 2 was 45.9 of rutin. It was confirmed to be equivalent to mg. On the other hand, the total flavonoid content of each 1 g of the Gujeolcho leaf extract of Comparative Example 1 and the Gujeolcho stem extract of Comparative Example 2 was equal to 11.3 mg and 5.8 mg of each rutin, and it was confirmed that the total flavonoid content was lower than that of Examples 1 and 2. I did.

That is, it was confirmed that the Gujeolcho flower extract of Example 1 and the Gujeolcho stem extract of Example 2 had higher antioxidant efficacy and higher total polyphenol and total flavonoid content compared to the extracts of Comparative Examples 1 and 2. Particularly, among the extracts of Examples 1 and 2, it was confirmed that the Gujeolcho stem extract of Example 2 has higher antioxidant efficacy and higher total polyphenol and total flavonoid content than the Gujeolcho flower extract of Example 1.

Test Example 3. NO (nitric oxide) measurement_anti-inflammatory

It was intended to measure the ability to produce nitric oxide, which is an inflammatory reaction result, induced by treatment of LPS (lipopolysaccharide) in RAW264.7 cells of Gujeolcho extract prepared in Examples and Comparative Examples. Macrophage RAW264.7 was cultured in DMEM medium supplemented with 10% fetal bovine serum at 37°C. The cultured cells were cultured in a 48-well plate at a concentration of 5×10 ⁴ cells/well and cultured to a density of about 90%. After replacing with DMEM medium without fetal bovine serum and incubating for 6 hours, dilute the hot water extract with DMEM medium without fetal bovine serum and phenol red to concentrations of 100, 200, 400 and 800 μg/mL, respectively, and then again in 48-well medium. After removal, it was replaced with a diluted extract. After 1 hour incubation, each well was treated with LPS at a concentration of 200 ng/mL and incubated for 18 hours.After 50 μL of the culture supernatant was transferred to a new 96-well plate, a solution of sulfanilamide solution and NED solution in a 1:1 ratio (Griess Reagent System, Promega G2930) 50 μL each, reacted for 15 minutes, and absorbance was measured at 540 nm. The control group was treated with DMEM medium without fetal bovine serum and phenol red instead of LPS treatment. The LPS control group was treated with DMEM medium without fetal bovine serum and phenol red instead of hot water extract treatment. The amount of nitric oxide produced was calculated by substituting NaNO ₂ into the standard curve prepared by reacting the sulfanilamide solution and the NED solution in a 1:1 ratio by concentration.

In the inflammatory process in the body, inflammatory factors such as excess nitric oxide (NO) and prostaglandin E ₂ (PGE ₂ ) are formed by induced NO synthase (iNOS) and cycloxygenase (COX)-2. Among them, NO has various physiological functions such as defense function, signaling function, neurotoxicity, and vasodilation in the body. NO itself has a very short half-life of about 6 to 10 seconds, and nitric oxide syntase (NOS) that forms NO is divided into three types of homologous enzymes such as type I, Ⅱ and Ⅲ according to physicochemical properties. Type Ⅰ (neuronal NOS, nNOS) and type Ⅲ (endothelial, eNOS) are classified as constitutive NOS (constitutive NOS) because they exist continuously in cells. It is divided into type III induced NOS (iNOS), which is expressed only when exposed. These NOSs formed NO by converting l-argine to l-citrulline. These NOSs are expressed by iNOS absolutely high, which played an important pathological action. In general, NO formation plays an important role in killing bacteria or removing tumors, but excessive NO formation due to pathological causes induces inflammation, causing tissue damage, genetic mutation, and nerve damage.

division Control LPS (ng/ml) 0 μg/ml 100 μg/ml 200 μg/ml 400 μg/ml 800 μg/ml Example 1 4.3 33.4 32.1 29.8 24.7 18.4 Example 2 6.1 33.6 30.1 28.8 22.9 15.4 Comparative Example 1 4.1 41.4 40.1 39.0 38.5 38.0 Comparative Example 2 4.4 43.8 43.5 42.5 42.1 41.4

As shown in Table 3 above, it was confirmed that the production of nitric oxide was suppressed as both the Gujeolcho flower extract prepared according to Example 1 of the present invention and the Gujeolcho stem extract prepared according to Example 2 increased to 800 μg/mL concentration. I did. On the other hand, it was confirmed that the extracts of Comparative Examples 1 and 2 did not suppress the amount of nitric oxide production compared to the extracts of Examples 1 and 2.

In the case of the Gujeolcho flower extract of Example 1, it was confirmed that the 800 μg/mL treatment significantly decreased when compared to the LPS-treated group, and the Gujeolcho stem extract of Example 2 was significantly greater than 400 μg/mL. It was confirmed that it was reduced to.

That is, it was confirmed that the Gujeolcho stem extract of Example 2 has superior anti-inflammatory activity compared to the Gujeolcho flower extract of Example 1.

These results indicate that when considering the results of ABTS antioxidant efficacy evaluation, total polyphenol content and total flavonoid content measurement, the Gujeolcho stem extract of Example 2 has higher antioxidant efficacy compared to the Gujeolcho flower extract of Example 1 and total polyphenol and total flavonoid content. It seems to be due to this high.

Hereinafter, examples of the formulation of the composition containing the powder of the present invention will be described, but the present invention is not intended to limit it, but is intended to be described in detail.

Formulation Example 1. Preparation of powder

500 mg of mixed extract powder obtained in Example 1

100 mg lactose

10 mg of talc

The above ingredients are mixed and filled in an airtight cloth to prepare a powder.

Formulation Example 2. Preparation of tablet

300 mg of mixed extract powder obtained in Example 1

100 mg corn starch

100 mg lactose

2 mg of magnesium stearate

After mixing the above ingredients, tablets are prepared by tableting according to a conventional tablet preparation method.

Formulation Example 3. Preparation of Capsule

200 mg of mixed extract powder obtained in Example 1

3 mg of crystalline cellulose

14.8 mg lactose

Magnesium stearate 0.2 mg

According to a conventional capsule preparation method, the above ingredients are mixed and filled into gelatin capsules to prepare a capsule.

Formulation Example 4. Preparation of injection

600 mg of mixed extract powder obtained in Example 1

Mannitol 180 mg

2974 mg of sterile distilled water for injection

Na ₂ HPO _4, 12H ₂ O 26 mg

It is prepared with the above ingredients per ampoule according to a conventional injection preparation method.

Formulation Example 5. Preparation of liquid formulation

4 g of mixed extract powder obtained in Example 1

10 g of isomerized sugar

5 g of mannitol

Purified water appropriate amount

According to the usual preparation method of liquid preparation, add each ingredient to purified water to dissolve it, add lemon zest, mix the above ingredients, add purified water and add purified water to adjust the total to 100g, then fill in a brown bottle for sterilization. To prepare a solution.

Formulation Example 6. Preparation of granules

1,000 mg of mixed extract powder obtained in Example 1

Vitamin mixture right amount

Vitamin A acetate 70 ㎍

1.0 mg of vitamin E

Vitamin B1 0.13 mg

Vitamin B2 0.15 mg

0.5 mg of vitamin B6

Vitamin B12 0.2 ㎍

Vitamin C 10 mg

Biotin 10 ㎍

1.7 mg of nicotinic acid amide

Folic acid 50 ㎍

0.5 mg of calcium pantothenate

Suitable amount of inorganic mixture

1.75 mg ferrous sulfate

Zinc oxide 0.82 mg

Magnesium carbonate 25.3 mg

Potassium monophosphate 15 mg

Dicalcium phosphate 55 mg

90 mg of potassium citrate

100 mg of calcium carbonate

Magnesium chloride 24.8 mg

The composition ratio of the vitamin and mineral mixture is relatively suitable for granules, but it is possible to arbitrarily modify the mixing ratio. After mixing the above ingredients according to a conventional granule preparation method, granules And can be used to prepare a health functional food composition according to a conventional method.

Formulation Example 7. Preparation of functional beverage

1,000 mg of mixed extract powder obtained in Example 1

1,000 mg citric acid

100 g oligosaccharides

2 g of plum concentrate

1 g taurine

Total 900 mL with purified water

After mixing the above ingredients according to a normal health drink manufacturing method, stirring and heating the resulting solution at 85°C for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed and sterilized, and stored in a refrigerator. It is used to prepare the functional beverage composition of the invention.

The composition ratio is composed of ingredients suitable for a relatively preferred beverage in a preferred embodiment, but the mixing ratio may be arbitrarily modified according to regional and ethnic preferences, such as the demand class, the country of demand, and the purpose of use.

In addition, a formulation example of a cosmetic composition suitable for applying the present invention will be presented.

Formulation Example 8. Lotion

Examples of the preparation of lotion among the cosmetics containing the extract of Example 1 are shown in Table 4 below.

ingredient Content (% by weight) Extract of Example 1 5.0 glycerin 6.0 1,3-butylene glycol 3.0 PEG 1500 1.0 Allantoin 0.1 DL-panthenol 0.3 EDTA-2Na 0.02 Benzophenone-9 0.04 Sodium hyaluronate 5.0 ethanol 10.0 Polysorbate 20 0.2 Preservatives, fragrances, and pigments a very small amount Distilled water Balance Sum 100

Formulation example 9. Lotion

Examples of the preparation of lotions among cosmetics containing the extract of Example 1 are shown in Table 5 below.

ingredient Content (% by weight) Extract of Example 1 3.0 Propylene glycol 6.0 glycerin 4.0 Triethanolamine 1.2 Tocopheryl Acetate 3.0 Liquid paraffin 5.0 Squalane 3.0 Macadamia Nut Oil 2.0 Polysorbate 60 1.5 Sorbitansquioleate 1.0 Carboxyvinyl polymer 1.0 Preservatives, fragrances, and pigments a very small amount Distilled water Balance Sum 100

Formulation example 10. Nourishing Cream

Examples of the preparation of a nourishing cream among cosmetics containing the extract of Example 1 are shown in Table 6 below.

ingredient Content (% by weight) Extract of Example 1 1.0 Lipotype glycerin monostearate 1.5 Cetearyl alcohol 1.5 Stearic acid 1.0 Polysorbate 60 1.5 Sorbitan Stearate 0.6 Isostearyl isostearate 5.0 Squalane 5.0 Mineral oil 35.0 Dimethicone 0.5 Hydroxyethyl cellulose 0.12 glycerin 6.0 Triethanolamine 0.7 Preservatives, fragrances, and pigments a very small amount Distilled water Balance Sum 100

Formulation example 11. Essence

Examples of the preparation of the essence among the cosmetics containing the extract of Example 1 are shown in Table 7 below.

ingredient Content (% by weight) Extract of Example 1 1.5 glycerin 10.0 Betaine 5.0 PEG 1500 2.0 Allantoin 0.1 DL-panthenol 0.3 EDTA-2Na 0.02 Benzophenone-9 0.04 Hydroxyethyl cellulose 0.1 Sodium hyaluronate 8.0 Carboxyvinyl polymer 0.2 Triethanolamine 0.18 Octyldodecanol 0.3 Octyldodeces-16 0.4 ethanol 6.0 Preservatives, fragrances, and pigments a very small amount Distilled water Balance Sum 100

Formulation example 12. Mask For pack latex

Among the cosmetics containing the extract of Example 1, an example of preparing an emulsion for a mask pack is shown in Table 8 below.

ingredient Content (% by weight) Extract of Example 1 1.0 Polyvinyl alcohol 15.0 Cellulose gum 0.15 glycerin 3.0 PEG 1500 2.0 Cyclodextrin 0.15 DL-panthenol 0.4 Allantoin 0.1 Monoammonium glycyrrhizinate 0.3 Nicotinamide 0.5 ethanol 6.0 PEG 40 hydrogenated castor oil 0.3 Preservatives, fragrances, and pigments a very small amount Distilled water Balance Sum 100

Claims (4)

A pharmaceutical composition for the prevention or treatment of inflammation, characterized in that it contains a Gujeolcho flower extract or a Gujeolcho stem extract as an active ingredient. The pharmaceutical composition for preventing or treating inflammation according to claim 1, wherein the Gujeolcho stem extract is an extract obtained by mixing Gujeolcho stems and leaves in a weight ratio of 1:0.1-0.8. Health functional food for preventing or improving inflammation, characterized in that it contains Gujeolcho flower extract or Gujeolcho stem extract as an active ingredient. A cosmetic composition having anti-inflammatory or antioxidant activity, characterized in that it contains Gujeolcho flower extract or Gujeolcho stem extract as an active ingredient.