KR20110023435A - Composition containing glycoproteins extract from plant - Google Patents

Abstract

PURPOSE: A composition containing plant glycoprotein extract is provided to prevent or treat wrinkle and inflammation and to improve moisturizing and skin regeneration. CONSTITUTION: A composition for preventing or treating skin wrinkle contains 0.1-30 weight% of glycoprotein as an active ingredient, isolated from Bergenia cordifolia, Rhodiola rosea, Comarum palustre, Abies Sibirica, Betula pendula Buds, or Inonotus Obliquus. A method for extracting the glycoprotein comprises: a step of chopping the plants and mixing with organic acid solution of pH 2-3 to obtain an aqueous solution; a step of adding protease and treating at 30-50°C for 2-24 hours; a step of filtering glycoprotein solution and raising temperature to 85°C; a step of continuously heating for predetermined time for sterilization; and a step of centrifuging to remove foreign materials and drying.

Description

Composition containing glycoproteins extract from plant}

The present invention relates to a composition containing a glycoprotein extracted from a plant.

Recently, the average life expectancy of human beings is getting longer, not how long they live, but how long they stay young. As you age, aging is an inevitable phenomenon. In addition, in modern societies, where pollution, stress are getting worse and UV exposure is becoming more frequent, aging is beginning earlier than before.

All the organs of the human body are gradually aging at a certain rate. Among them, skin aging proceeds under the influence of various natural aging phenomena, which is a complex process. Most of the early skin aging begins with the drying of the skin, specifically, the decrease in elasticity and wrinkles caused by the lack of moisture. As you age, your cells lose their regeneration and lose their elasticity. Wrinkles are formed by the growth of connective tissue on the skin that is destroyed by the effects of hormones and UV rays.

Specific skin aging effects include slow skin regeneration, aging dead skin cells, rough skin, wrinkles due to reduced collagen synthesis, and poor skin protection, pigmentation such as spots and spots. In addition, the skin's protective function is lowered, and it reacts easily to drugs and irritation.

The present invention seeks to provide a composition for preventing or improving skin wrinkles, skin whitening, inflammation prevention or improvement, skin moisturization, skin regeneration, scar prevention or improvement, atopic skin improvement, antioxidant enhancement, anti aging or improvement.

The present invention to achieve the above object is Bergenia cordifolia ( Bergenia cordifolia ), Rhodiola ( Rhodiola) rosea ), Comarum palustre , Abies Sibirica Sibirica , Betula pendula Buds ), and Inonotus Obliquus provides a composition containing as an active ingredient glycoprotein extracted from at least one selected from the group consisting of.

In one embodiment of the present invention, the composition is characterized in that it has a purpose of preventing or improving skin wrinkles.

In one embodiment of the present invention, the composition is characterized in that it has a skin whitening use.

In one embodiment of the present invention, the composition is characterized in that it has a purpose of preventing or ameliorating inflammation.

In one embodiment of the invention, the composition is characterized in that it has a skin moisturizing use.

In one embodiment of the present invention, the composition is characterized in that it has a purpose of promoting skin regeneration.

In one embodiment of the present invention, the composition is characterized in that it has a use for preventing or improving scars.

In one embodiment of the present invention, the composition is characterized in that it has an atopic skin improvement use.

In one embodiment of the present invention, the composition is characterized in that it has an antioxidant enhancement use.

In one embodiment of the present invention, the composition is characterized in that it has an anti-aging or improving use.

The composition of the present invention comprises a glycoprotein extracted from at least one selected from the group consisting of bergenia cordifolia, rhodiola rossia, comarum parustre, abis civicica, betula pendula buzz, and inototus obricus. It is contained as an active ingredient, and has the effect of preventing or improving skin wrinkles, skin whitening, inflammation prevention or improvement, skin moisturizing, promoting skin regeneration, preventing or improving scars, improving atopic skin, enhancing antioxidants, preventing or improving aging.

As used herein, the term "extract" includes any material obtained by extracting a component therefrom from a natural product, regardless of the method of extraction or the kind of the component. For example, it is a broad concept that includes all components obtained by extracting a component dissolved in a solvent from a natural product using water or an organic solvent, and those obtained by extracting only a specific component such as an oil such as oil.

As used herein, the term "skin" refers to a tissue covering the body surface of an animal, and is a broad concept including not only tissues covering the body surface such as the face or body, but also the scalp and hair.

Hereinafter, the present invention will be described in detail.

One embodiment of the present invention is a sugar extracted from one or more selected from the group consisting of bergenia cordifolia, rhodiola roscia, comarum parustre, abis civicica, betula pendula buzz, and innoteus obricus A composition containing a protein as an active ingredient.

Bergenia cordifolia , also known as Siberian rock odor , is a perennial herb of the genus Rosaceae. It is native to Siberia and Mongolia. It is about 30-40 cm tall, and the flower blooms reddish purple or light pink, rarely white, in late spring or early spring. Rhodiola rosea ( Rhodiola rosea ) is a genus of locusts, a stone tree family, also known as Honggyeongcheon, and grows mainly in mountains near the Arctic in Europe or Asia. Comarum palustre ), also known as chabilnik balotni , is a herb belonging to the Rosaceae family and growing mainly in Russia. Abies Sibirica is an Siberian fir belonging to the pine tree Pineaceae, growing mainly in the humid regions of the mountains in the cold north. Betula pendula belongs to the oak tree birch family, also known as pendula birch and silver birch, mainly using snow. Inhabits mountains throughout Europe and Asia and grows at high altitudes in Southern Europe. Inonotus Obliquus belongs to the pine scale family, also known as chaga . It is native to Siberia, and is native to Siberia, birch trees of more than 45 degrees north latitude, such as North America and Northern Europe.

As used herein, "collagen" refers to an important fibrous protein of animal connective tissue, which has the functions of preventing or improving skin wrinkles, regenerating cells, preventing or improving scars, strengthening skin elasticity, and preventing skin aging. "Procollagen" is a precursor of collagen found in the skin and other organs of various vertebrates, resulting in the same function as collagen. Thus, elevated levels of collagen and procollagen may prevent or improve skin wrinkles, regenerate cells, prevent or improve scars, enhance skin elasticity, and prevent or improve skin aging. Meanwhile, "collagenase" is an enzyme that breaks down collagen and is known as one of the causes of skin wrinkles. Therefore, by inhibiting its expression, the collagen level of the skin is increased, so skin wrinkles can be prevented or improved, cells are regenerated, skin elasticity is strengthened, and skin aging can be prevented or improved. The compositions disclosed herein are Bergenia cordifolia, Rhodiola rossia, Komarum parustre, Abis sibirica, Vesica, which have the effect of promoting procollagen biosynthesis and inhibiting the production of collagenase (MMP-1). It contains glycoproteins extracted from one or more selected from the group consisting of Tula Pendula Buzz and Innoteus Olivus as active ingredients to prevent or improve skin wrinkles, regenerate cells, prevent or improve skin wrinkles, strengthen skin elasticity and prevent skin aging Can be used as

"Melanin" is a black-brown pigment, which has a function of blocking a certain amount of ultraviolet rays to protect the skin from ultraviolet rays, so when the skin is exposed to ultraviolet rays, a large amount of melanin is produced in the body to protect the skin. At this time, since the skin color is determined by the amount of melanin, the more the amount of melanin becomes black, which causes blemishes, spots, freckles, and mushrooms. Therefore, the composition that inhibits the production of melanin may have a skin whitening effect by preventing the skin from becoming black. The composition disclosed herein is a group consisting of bergenia cordifolia, rhodiola roscia, comarum parustre, abis civicica, betula pendula buzz, and innoteus oblicus, which have an effect of inhibiting melanogenesis. It can be used for skin whitening by containing the glycoprotein extracted from one or more selected from as an active ingredient.

Interleukin-6 (IL-6) is a cytokine, also called B cell stimulating factor 2 (BSF2) or interferon β2 (INF-β2), and is known to mediate the inflammatory response in inflammatory diseases. Interleukin-8 (IL-8), also called neutrophil attracting / activating protein-1 (NAP-1), is also known to mediate the inflammatory response and induce histamine release in atopic dermatitis patients. MCP-1 (Momocyte Chemoattractant Protein-1), also called CCL-2, is also known as an inflammatory cytokine. Therefore, when the biosynthesis of the cytokines is inhibited, it may have an anti-inflammatory action and further prevent or improve atopic skin. The compositions disclosed herein are Bergenia cordifolia, Rhodiola rossia, Komarum parustre, which have the effect of inhibiting LPS-induced inflammation and inhibiting the biosynthesis of IL-6, IL-8, and MCP-1. Contains, as an active ingredient, glycoproteins extracted from one or more selected from the group consisting of Abis Sibirica, Betula Pendula Buzz, and Innoteus Olivus, to prevent or improve inflammation, improve atopic skin, and prevent skin aging Or for improvement purposes.

PPARγ is a factor generally present in skin constituent cells and maintains homeostasis of skin barrier function by regulating epidermal keratinocyte differentiation and lipid metabolism to induce normal epidermal layer formation. It is known to play an important role in the manifestation of the healing process. Due to these characteristics, PPARγ can act as a key regulator of various skin diseases such as psoriasis, wound healing and acne due to overgrowth of the epidermal layer as well as inflammation-related skin diseases. In particular, when PPARγ is activated, it can be expected to moisturize the skin by strengthening the skin barrier function. The compositions disclosed herein are in the group consisting of Bergenia cordifolia, Rhodiola rossia, Comarum parustre, Avis civicica, Bettula pendula buzz, and Innoteus oblicus, which have the effect of activating PPARγ. By containing glycoprotein extracted from one or more selected ingredients as an active ingredient, it has a skin moisturizing effect, improves atopic skin, and further prevents or improves skin aging.

As used herein, "cell regeneration" refers to the phenomenon of regenerating cells of damaged parts, and "skin cell regeneration" particularly means regenerating skin cells. If skin regeneration is not performed smoothly, aging of the skin is promoted, and melanin produced in the pigment cells in the epidermis is not discharged, and there is pigmentation remaining. On the other hand, if the skin is regenerated smoothly, it can prevent or improve skin scars and help moisturize the skin. The compositions disclosed herein include Vergenia cordifolia, Rhodiola roscia, Comumarum parustre, Abis sibirica, Bettula pendula buzz, and Innoteus Aubrey, which have the effect of promoting cell regeneration, particularly skin regeneration. By containing glycoprotein extracted from at least one selected from the group consisting of couss as an active ingredient, it prevents or improves skin wrinkles, has a skin whitening and skin moisturizing effect, promotes skin regeneration, prevents or improves scars, and further aging Can be prevented or improved.

If there is more oxygen in the body than necessary, some of the remaining oxygen becomes free radicals. Reactive free radicals attack cells and damage them. Oxidized oxygen, which attacks biological tissues and damages cells, is called free radicals, which are generally produced by the body's metabolic processes and are abnormally increased by oxygen intake, stress, and other causes when breathing. These free radicals have strong cytotoxicity and need to be removed. As used herein, "antioxidant" refers to the action of removing free radicals that are harmful oxygen. The compositions disclosed herein are at least one selected from the group consisting of antioxidants Bergenia Cordifolia, Rhodiola Roscia, Comarum Parustre, Abis Sibirica, Bettula Pendula Buzz, and Innotus Olivus By containing the glycoprotein extracted from as an active ingredient, it is possible to prevent damage to cells caused by active oxygen or ultraviolet rays, and to prevent or improve skin wrinkles, thereby preventing or improving aging.

In one embodiment of the present invention sugars extracted from one or more selected from the group consisting of bergenia cordifolia, rhodiola roscia, comarum parustre, abis sibirica, betula pendula buzz, and innoteus obricus The protein may be contained in 0.1 to 30% by weight relative to the total composition weight, depending on the form of the composition. If the content is less than 0.1% by weight, the effect is insignificant. If the content is more than 30% by weight, there may be a problem in composition stability.

In one embodiment according to the present invention the effect of the present invention is composed of bergenia cordifolia, rhodiola roscia, comarum parustre, abis civicica, betula pendula buzz, and innoteus obliques. It comes from glycoproteins extracted from one or more selected from the group.

The method of extracting glycoproteins from each of the plants is as follows: After chopping the refrigerated raw plants with a shredder, the organic acid solution of pH 2 to 3 is mixed at a ratio of 1: 2 to 1: 5 based on the weight of the raw materials, After preparing the temperature is maintained at 30 ~ 50 ℃. While adjusting the pH of the aqueous solution to 1.0 to 3.0, 0.1g% to 1g% by weight of the proteolytic enzyme is added to the organic acid aqueous solution and treated for 2 hours to 24 hours at a temperature of 30 to 50 ℃. When the concentration of the enzyme-treated aqueous solution reaches the desired concentration, the first filtration process is performed to filter only the glycoprotein aqueous solution. The reaction is terminated by sterilizing simultaneously with deactivation of the enzyme by rapidly raising the temperature of the filtered aqueous solution to 85 ° C. or higher and continuing heating for a predetermined time. The sterilized extract is then removed using a centrifugal separator and dried using a drying apparatus to complete the glycoprotein powder product (see Korean Patent Publication No. 10-2004-27283).

The glycoprotein may also be obtained by a general method of leaching, purifying and separating each plant raw material.

The present invention comprises a glycoprotein extracted from at least one selected from the group consisting of bergenia cordifolia, rhodiola rossia, comarum parustre, abis civicica, betula pendula buzz, and innotus obliques. A cosmetic composition comprising the composition can be provided. The cosmetic composition may for example be a cosmetic, whereby the cosmetic appearance contains a cosmetically or dermatologically acceptable medium or base. These are all formulations suitable for topical application, for example, emulsions obtained by dispersing the oil phase in solutions, gels, solids, pasty anhydrous products, aqueous phases, emulsions obtained by dispersing the aqueous phase in oil phases, suspensions, microemulsions, microcapsules, microcapsules. It may be provided in the form of granulocytes or ionic (liposomal) and nonionic vesicle dispersants, or in the form of creams, skins, lotions, powders, ointments, sprays, cleansers or cone sticks. These compositions may be prepared according to conventional methods in the art. The composition according to the invention can also be used in the form of a foam or in the form of an aerosol composition further containing a compressed propellant.

Cosmetic compositions may include fatty substances, organic solvents, solubilizers, thickeners, gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, ionic or nonionic emulsifiers, fillers Cosmetic or dermatology such as metal ion sequestrants, chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic actives, lipid vesicles or any other ingredients commonly used in cosmetics It may contain adjuvants commonly used in the art. Such adjuvants are introduced in amounts generally used in the cosmetic or dermatological arts.

The cosmetic composition is not particularly limited in the formulation, for example, supple cosmetics, astringent cosmetics, nourishing cosmetics, nutrition cream, massage cream, essence, eye cream, eye essence, cleansing cream, cleansing foam, cleansing water, It can be formulated into cosmetics such as packs, powders, body lotions, body creams, body oils and body essences.

In addition, the present invention provides a glycoprotein extracted from at least one selected from the group consisting of Bergenia cordifolia, Rhodiola rossia, Komarum parustre, Avis civicica, Betula pendula buzz, and Innotus obliques. It provides a pharmaceutical composition comprising a composition containing. Such pharmaceutical compositions may further contain pharmaceutical adjuvants such as preservatives, stabilizers, hydrating or emulsifying accelerators, salts and / or buffers for controlling osmotic pressure, and other therapeutically useful substances, and may be prepared in various oral forms according to conventional methods. It may be formulated in the form of a dosage form or parenteral dosage form.

The oral dosage forms include, for example, tablets, pills, hard and soft capsules, solutions, suspensions, emulsifiers, syrups, powders, powders, fine granules, granules, pellets, and the like. Active agents, diluents (e.g. lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and glycine), glidants (e.g. silica, talc, stearic acid and its magnesium or calcium salts and polyethylene glycols) have. Tablets may also contain binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and polyvinylpyrrolidine, optionally starch, agar, alginic acid or its sodium salt Pharmaceutical additives such as disintegrants, absorbents, colorants, flavors, and sweeteners. The tablets can be prepared by conventional mixing, granulating or coating methods.

In addition, the parenteral administration agent may be, for example, in the form of injections, drops, ointments, lotions, gels, creams, sprays, suspensions, emulsions, suppositories, patches, and the like, but is not limited thereto. It is not.

The pharmaceutical composition according to the present invention may be administered orally, parenteral, rectal, topical, transdermal, intravenous, intramuscular, intraperitoneal, subcutaneous.

In addition, the dosage of the active ingredient will vary depending on the age, sex and weight of the subject to be treated, the specific disease or pathology to be treated, the severity of the disease or pathology, the route of administration and the judgment of the prescriber. Dosage determination based on these factors is within the level of skill in the art. Typical dosages are 0.001 mg / kg / day to 2000 mg / kg / day, more specifically 0.5 mg / kg / day to 2.5 mg / kg / day.

The present invention comprises a glycoprotein extracted from at least one selected from the group consisting of bergenia cordifolia, rhodiola rossia, comarum parustre, abis civicica, betula pendula buzz, and innotus obliques. It provides a health food composition comprising the composition. The health food composition is not particularly limited in the formulation, for example, it may be formulated as a tablet, granules, drinks, caramel, diet bar and the like. In addition to the active ingredient, the health food composition of each formulation may be suitably selected by a person skilled in the art according to the formulation or purpose of use, in addition to the active ingredient, and may be synergistic when applied simultaneously with other raw materials. .

Determination of the dosage of the active ingredient is within the level of those skilled in the art, and the daily dosage of the drug depends on various factors such as less progression, onset, age, health condition, complications, etc. of the subject to be administered. Generally, the composition may be administered by dividing the composition 1 to 500 mg / kg, preferably 30 to 200 mg / kg, once or twice a day, and the dosage may be in the range of the present invention by any method. It is not limiting.

Hereinafter, the configuration and effects of the present invention will be described in more detail with reference to Examples and Test Examples. However, these examples and test examples are provided only for the purpose of illustration in order to facilitate understanding of the present invention, and the scope and scope of the present invention is not limited by the following examples and test examples.

[Example]

Chilled citric acid at pH 2 to 3 after chopping each of the refrigerated bergenia cordifolia, rhodiola roscia, comarum parustre, abis civicica, betula pendula buzz, and innoteus obliques The aqueous solution was mixed with the chopped plant in a ratio of 1: 3 to prepare an aqueous solution. While adjusting the pH of the aqueous solution to 2.0, 0.3 g% by weight of pepsin was added to the aqueous solution and treated at 40 ° C. for 18 hours.

When a 5% yield of glycoprotein was obtained compared to the enzyme-treated aqueous solution, the temperature was rapidly increased to 85 ° C. and treated for 40 minutes. After that, the obtained extract was centrifuged to remove foreign substances, thereby preparing each glycoprotein in liquid form. Glycoproteins obtained from Bergenia cordifolia, Rhodiola rossia, Comarum parustre, Avis civicica, Bettula pendula buzz, and Innotus oblicus were prepared in Examples 1, 2 and 3, respectively. It was set as Example 4, Example 5, and Example 6.

Examples obtained through the extraction and separation process as described above had an amino acid composition as shown in Table 1 below.

Example 1 Example 2 Example 3 Example 4 Example 5 Example 6 Aspartic acid 45.11 4.20 10.94 10.50 6.24 9.82 Clutamic Acid 21.59 62.25 33.12 24.38 14.91 14.98 Serine 2.39 1.07 3.49 5.27 2.31 3.67 Glycine 3.00 0.81 2.66 4.24 3.10 2.36 Histidine 1.37 0.43 1.22 1.75 0.68 0.41 Arginine 2.44 3.59 5.33 4.12 0.98 0.39 Threonine 1.09 0.39 1.52 2.75 1.28 2.00 Alanine 5.75 1.14 10.55 7.65 2.54 2.59 Proline 12.72 25.18 26.54 33.93 61.09 58.45 Tyrosine 0.58 0.19 0.55 0.28 1.04 0.38 Balin 1.35 0.13 1.25 1.82 1.33 1.64 Methionine 0.23 0.05 0.20 0.21 0.59 0.28 Cysteine 0.14 0.11 0.23 0.01 0.06 0.05 Isoleucine 0.55 0.15 0.88 0.92 0.74 0.87 Leucine 0.70 0.13 0.76 1.33 1.15 1.36 Phenylalanine 0.43 0.08 0.43 0.51 0.92 0.53 Lysine 0.58 0.11 0.34 0.33 1.04 0.22 synthesis 100 100 100 100 100 100

Test Example 1 Procollagen Biosynthesis

Fibroblasts isolated from human skin were seeded at 10 ⁶ per hole in 24 holes and cultured until 90% growth. After incubation in serum-free medium for 24 hours, the examples were each dissolved in serum-free medium, treated at 100 ppm, and incubated in a CO ₂ incubator for 24 hours. The supernatant was removed and procollagen was increased or decreased using the procollagen type (I) Elisa kit (Procollagen type 1). Vitamin C was also tested as a positive control. The test was carried out at a concentration in consideration of cytotoxicity, and the results are shown in Table 2. Synthetic capacity is prepared by using an untreated group as 100.

ingredient Synthetic Performance (%) Untreated group 100 Vitamin c 128.5 Example 1 122.8 Example 2 123.4 Example 3 117.6 Example 4 119.4 Example 5 126.2 Example 6 107.3

As can be seen in Table 2, the procollagen synthesis ability of the examples was superior to the untreated group. Therefore, the embodiments can alleviate collagen reduction caused by skin aging by promoting collagen production. Therefore, the composition containing the glycoprotein extracted from each plant can prevent or improve skin wrinkles, prevent or improve scars, and prevent or improve skin aging. And glycoproteins extracted from each plant can be used by mixing two or more kinds.

Test Example 2 Collagenase (MMP-1) Inhibitory Activity

The test was performed using a 96-well microtiter plate containing Dulbecco's Modified Eagle's Medium (DMEM) containing 2.5% fetal bovine serum. well) and incubated until 70 ~ 80% growth. And the Examples were treated for 24 hours at a concentration of 10ppm, and then the cell culture was filled. The collected cell culture was measured for collagenase expression using a commercially available collagenase measuring instrument (Amersham Pharmacia Co., Catalog #: RPN 2610). First, the cell culture medium was placed in a 96-hole plate uniformly coated with primary collagenase antibody, and the antigen-antibody reaction was performed in a thermostat for 3 hours. After 3 hours, the chromophore-conjugated secondary collagen antibody was placed in a 96-hol plate and reacted again for 15 minutes. After 15 minutes, a color causing substance was added to cause color development at room temperature for 15 minutes, and 1M sulfuric acid was added again to stop the reaction (coloring), thereby making the reaction solution yellow. At this time, the degree of yellow appears differently depending on the progress of the reaction. The absorbance of the yellowish 96-coated plate was measured at 405 nm using an absorbance meter, and the expression of collagenase was expressed by Equation 1 below based on the reaction absorbance of the untreated group cultured cell culture medium without any treatment. The degree was calculated. A positive control treated only with retinoic acid and a negative control treated only with UV were tested together. The test was conducted at the concentration considering the cytotoxicity, and the results are shown in Table 3.

Collagenase expression level (%) = (absorbance of each test substance / absorbance of untreated group) X 100

ingredient Collagenase expression suppression degree (%) Untreated group 100 UV treatment 151.2 Retinoic acid 73.5 Example 1 38.6 Example 2 21.3 Example 3 40.4 Example 4 86.7 Example 5 75.8 Example 6 67.3

As the inhibition of collagenase expression increases, the percentage decreases. As shown in Table 3, each of the examples has an effect of inhibiting collagenase matrix metalloprotease (MMP-1). That is, the composition containing the glycoprotein extracted from each plant may inhibit the biosynthesis of the skin tissue degrading enzyme MMP-1 generated by internal aging or external environmental factors, thereby inhibiting collagen reduction in the skin and improving wrinkles. And glycoproteins extracted from each plant can be used by mixing two or more kinds.

Test Example 3 Evaluation of Melanin Inhibitory Activity Using Melan-a Cells

Melan-a cells (obtained from St. George's Hospital Medical School, UK) were plated in DMEM containing 10% FBS in 6-hole plates (melanin quantitative), 2 X 10 ⁴ cells / ball (final volume 3 ml), 96-hole plates (MTT assay) was added 2 × 10 ³ cells / ball (final volume 200μl) and incubated for 24 hours in 37 ℃, 5% CO ₂ incubator (MCO-20AIC, Sanyo). After incubation, the medium was removed and treated with DMEM containing 10% FBS, 2 μM α-MSH (melanosite stimulating hormone), 2 mM theophylline as a fresh medium and the examples were added to the medium and treated daily for 3 days. . On day 5, cultured medium was removed from the 6-hole plate and treated with 0.25% Trypsin-EDTA (Sigma Chemical Co., Ltd.) solution to recover the cell pellet, transferred to a 1.5 ml test tube, and centrifuged at 10,000 rpm for 10 minutes to remove the supernatant. Removed. The obtained pellet was dried at 60 ° C., and 100 μl of 1N NaOH was added to dissolve the melanin in the cells. The solution was diluted with PBS and then absorbance was measured at 475 nm of ELISA reader to determine the inhibition rate of melanin production in the examples. The untreated group, which was not treated with anything, was used as a negative control, and the arbutin-treated group was used as a positive control. The whitening effect was measured by measuring the degree of inhibition of melanin production in the example compared to the melanin content in the untreated group. To calculate the melanin production inhibition rate according to the following equation 2 and the results are shown in Table 4.

% Melanogenesis inhibition = (absorbance of each test substance / absorbance of untreated group) X 100

Melanin production inhibitory effect ingredient density Melanin production inhibition rate (% control) Untreated group No additives - Arbutin 50 ppm 43.8% Example 1 25 ppm 21.5% 50 ppm 33.8% Example 2 25 ppm 31.5% 50 ppm 36.3% Example 3 25 ppm 20.6% 50 ppm 27.4% Example 4 25 ppm 43.8% 50 ppm 51.3% Example 5 25 ppm 37.5% 50 ppm 43.2% Example 6 25 ppm 17.3% 50 ppm 27.5%

As can be seen from Table 4, it was recognized that each of the examples has an inhibitory effect on melanin production, and this effect was excellent as the concentration of glycoprotein was higher. Through this, the glycoproteins extracted from each plant according to the present invention inhibited melanin production, so it was confirmed that the whitening effect was excellent. And glycoproteins extracted from each plant can be used by mixing two or more kinds.

Test Example 4 Anti-inflammatory Activity Measurement (Cell-Based Analysis)

Raw 246.7 macrophage cell line (available from the Korean Cell Line Bank) was added to DMEM, 10% FBS penicillin (100 IU / mL) and streptomycin (100 μg / mL), and then incubated at 37 ^° C and 5% CO ₂ incubator. Cells were grown, and 300 μl were dispensed at a density of 5 × 10 ⁵ cells / ml on a 48-hole plate, and after 24 hours, the examples were treated for each concentration. After 1 hour, 1 μg / ml of LPS, an inflammation-causing substance, was treated, and the amount of NO released in the medium after 18 hours was measured using a Greis reagent. A group treated with 10 μM of dexamethasone (dexamethasone) was used as a positive control of anti-inflammatory activity that inhibited the release of NO, and a group treated with only 1 μg / ml of LPS was used as a negative control. The amount of NO was determined in comparison with the measured value of the standard NaNO ₂ and the results are shown in Table 5 below.

ingredient Extract concentration (ppm) % Inhibition 1 μg / ml LPS + 10 μM dexamethasone - 100 LPS 1 µg / ml - 0 1 μg / ml LPS + Example 1 50 42.2 LPS 1 μg / ml + Example 2 50 31.6 1 μg / ml LPS + Example 3 50 39.1 LPS 1 μg / ml + Example 4 50 39.6 1 μg / ml LPS + Example 5 50 41.1 LPS 1 μg / ml + Example 6 50 29.3

As shown in Table 5, each example has the effect of inhibiting the LPS-induced inflammation by inhibiting NO production compared to the negative control. Therefore, it was confirmed that glycoproteins extracted from each plant had an effect of preventing or improving inflammation. And glycoproteins extracted from each plant can be used by mixing two or more kinds.

[Test Example 5] Confirmation of the inhibitory effect of IL-6, IL-8, MCP-1 secretion by stimulation in the keratinocyte line HaCaT

Test cells, keratinocytes (Cell name: HaCaT obtained from Heidelberg, Germany), 10% fetal bovine sereum (FBS, from Gibco, USA) and 1% penicillin-streptomycin (US) Approximately 2 × 10 ⁴ cell counts / column in 48 well plates using DMEM (Dulbecco's Modified Eagle Medium, Lonza, USA) containing Penicillin-Streptomycin, obtained from Gibco, USA After dispensing at a density, 37 ° C, 5% CO ₂ Incubated for 24 hours under conditions. After the culture was removed, the cells were washed once with 200 μl of phosphate buffered saline (PBS), and 200 μl of DMEM containing no fetal bovine serum was added and cultured for 24 hours. After treating the examples with DMEM containing 1% fetal bovine serum at the concentrations mentioned in Table 6, Table 7 and Table 10 for 10 minutes, respectively, Retinoic acid of 10μM concentration (Sigma-Aldrich, USA) and incubated for 24 hours. 50 μl of the culture was taken to evaluate the secretion inhibitory effect of these cytokines using IL-6, IL-8, and MCP-1 ELISA kit (obtained by BD pharmigen, USA). On the other hand, the group using 1 ppm of hydrocortisone instead of each example as a positive control, the group without any treatment as a negative control, the group treated with only retinoic acid alone was also tested. The cytokine secretion inhibitory effect is obtained by drawing a standard curve based on the absorbance of rh IL-6, rh IL-8, and rh MCP-1 (obtained from BD pharmigen, USA), to obtain a reaction formula between absorbance and standard substance. , By substituting the absorbance of the examples to obtain the secretion amount of these cytokines, and evaluated by the method of obtaining the stimulation relaxation rate by the following equation (3). In addition, the inhibition rate of the cytokine secretion of each example was evaluated by calculating the inhibition rate according to the following Equation 4 based on the difference in cytokine secretion between HaCaT cells not treated with retinoic acid and HaCaT cells treated with retinoic acid.

% Irritation relief = [1-(cytokine secretion of each test substance / cytokine secretion of retinoic acid alone)] x 100

% Inhibition = {1-[(cytokine secretion of each test substance-cytokine secretion of the negative control) / cytokine secretion of each test substance]} x 100

Tables 6, 7, and 8 below are test results associated with IL-6, IL-8, and MCP-1, respectively.

ingredient Extract concentration (ppm) IL-6 secretion (pg / ml) Stimulation Relief (%) % Inhibition Negative Control - 20.12 - - Retinoic acid (10 μm) - 78.55 - - Retinoic acid (10 μm) +
Hydrocortisone (1 ppm) - 22.58 71.25 89.10 Retinoic acid (10 μm) +
Example 1 100 23.62 69.92 85.18 Retinoic acid (10 μm) +
Example 2 100 27.45 65.05 73.29 Retinoic acid (10 μm) +
Example 3 100 38.27 51.26 52.57 Retinoic acid (10 μm) +
Example 4 100 28.13 64.18 71.52 Retinoic acid (10 μm) +
Example 5 100 29.55 62.38 68.08 Retinoic acid (10 μm) +
Example 6 100 34.62 55.92 58.11

ingredient Extract concentration (ppm) IL-8 secretion (pg / ml) Stimulation Relief (%) % Inhibition Negative Control - 128.5 - - Retinoic acid (10 μm) - 521.9 - - Retinoic acid (10 μm) +
Hydrocortisone (1 ppm) - 159.7 69.40 80.46 Retinoic acid (10 μm) +
Example 1 100 200.3 61.62 64.15 Retinoic acid (10 μm) +
Example 2 100 145.6 72.10 88.25 Retinoic acid (10 μm) +
Example 3 100 192.6 63.09 66.71 Retinoic acid (10 μm) +
Example 4 100 207.5 60.24 61.92 Retinoic acid (10 μm) +
Example 5 100 163.3 68.71 78.68 Retinoic acid (10 μm) +
Example 6 100 138.3 73.50 92.91

ingredient Extract concentration (ppm) MCP-1 secretion (pg / ml) Stimulation Relief (%) % Inhibition Negative Control - 638.83 - - Retinoic acid (10 μm) - 1406.24 - - Retinoic acid (10 μm) +
Hydrocortisone (1 ppm) - 710.68 49.46 90.64 Retinoic acid (10 μm) +
Example 1 100 835.32 40.59 74.40 Retinoic acid (10 μm) +
Example 2 100 763.21 45.73 83.70 Retinoic acid (10 μm) +
Example 3 100 782.05 44.39 81.68 Retinoic acid (10 μm) +
Example 4 100 738.24 47.50 86.53 Retinoic acid (10 μm) +
Example 5 100 724.51 48.48 88.17 Retinoic acid (10 μm) +
Example 6 100 789.38 43.87 80.92

As can be seen in Tables 6, 7, and 8 above, each example shows the effect of inhibiting the secretion of IL-6, IL-8, and MCP-1. Therefore, the glycoproteins extracted from each plant were found to have an anti-inflammatory effect by inhibiting the secretion of the cytokines, which are inflammation-inducing factors, thereby preventing or improving inflammation. And glycoproteins extracted from each plant can be used by mixing two or more kinds.

Test Example 6 Evaluation of PPARγ Efficacy

1) Cell Culture

Normal African green monkey kidney epithelial cells, CV-1 cells (available from ATCC) were obtained from DMEM (Dulbecco Modified) containing 10% Fetal Bovine Serum and 100 unit / ml penicillin-streptomycin. Eagle's Medium) and incubated at 37 ℃, 5% CO ₂ .

2) Plasmid transfection

The plasmid has a PPARγ plasmid expressing PPARγ and a PPARs Response Element (PPRE) that is activated by binding to PPARs under normal culture conditions, and then firefly luciferase acting as a reporter PPRE-Luc plasmids with genes and three β-gal plasmids used to correct for transfection were used.

Dispense CV-1 cells in a 24-hole plate at a concentration of 5 x 10 ⁴ and incubate for 18 to 24 hours. Reaction of the plasmids with a 1: 3 (μg: μl) mixture of DMEM and PEI (Polyethylenimine) at room temperature for 15 minutes After the plasmids were introduced into the cells by spraying the respective balls.

3) PPAR-γ activity measurement

After plasmids were introduced into the cells and incubated for 24 hours to stabilize, the examples were treated and incubated for another 24 hours. After incubation, the cells were washed twice with cold PBS, and the cells were destroyed by treating the cells with reporter lysis buffer (Promega) for 15 minutes. The cell eluate thus obtained was transferred to a micro tube and centrifuged at 12,000 rpm for 3 minutes at 4 ° C. to separate cell residues, thereby obtaining pure cell eluate.

PPAR- [gamma] activity of each Example was compared by mixing the luciferase substrate (Promega) in the cell eluate in a 2: 1 ratio and measuring the amount of light emitted by the luminometer. In addition, the cell eluate was treated with the same amount of 2X β-galactosidase enzyme assay buffer (Promega), reacted at 37 ° C. for 30 minutes, and then measured for absorbance at 420 nm. -Galactosidase activity was compared. The PPAR-γ activity of each example was divided by the activity of each beta-galactosidase to correct the difference according to the degree of plasmid introduction, and the correction value was evaluated using this sample t-test of Minitab for the significance of the activity. Meanwhile, TGZ (troglitazone), a PPAR-γ agonist, was tested together as a positive control.

ingredient density Activity (% control) Untreated group No additives 100% TGZ 5 μM 563% Example 1 50 ppm 233% Example 2 50 ppm 319% Example 3 50 ppm 180% Example 4 50 ppm 255% Example 5 50 ppm 384% Example 6 50 ppm 129%

As can be seen in Table 9, PPARγ activity of each example was superior to the untreated group. Therefore, the glycoproteins extracted from each plant was found to have excellent skin moisturizing effect by activating PPARγ. And glycoproteins extracted from each plant can be used by mixing two or more kinds.

Test Example 7 Evaluation of Cell Regeneration Efficacy

Keratinocytes, HaCaT (cell name: HaCaT obtained from: German Cancer Institute, Heidelberg, Germany) were added to a 96-well plate incubator at 10 ⁴ per ball, incubator at 30 ° C with DMEM containing 10% FBS (fetal bovine serum). Incubated for 24 hours. Then, the examples were replaced with DMEM containing 1% FBS contained in 0.5, 2.8ppm concentration, respectively, and then incubated for 48 hours. The WST-1 solution (Roche) was added 1/10 times and reacted in a 37 ° C. incubator for 2 hours. The cultured plate incubator each measured the color development at 440 nm. When the degree of color development of the non-treated group was 100, the degree of color development of each example was evaluated to quantify the degree of proliferation of the cells. In addition, the untreated group treated with nothing and the group treated with only 10% FBS was tested together, and the results are shown in Table 10 below.

ingredient density % Of cell proliferation Untreated group No additives 100% FBS 10% 188% Example 1 0.5 ppm 157% 2.8ppm 132% Example 2 0.5 ppm 133% 2.8ppm 124% Example 3 0.5 ppm 129% 2.8ppm 131% Example 4 0.5 ppm 144% 2.8ppm 139% Example 5 0.5 ppm 129% 2.8ppm 147% Example 6 0.5 ppm 125% 2.8ppm 109%

As can be seen in Table 10, each example was superior to the effect of proliferating cells compared to the untreated group. That is, the glycoproteins extracted from each plant was confirmed that the cell proliferation effect is effective to regenerate the cells. Therefore, glycoproteins extracted from each plant may have skin wrinkle improvement or prevention, skin whitening, skin moisturizing, cell regeneration, scar prevention or improvement, aging prevention or improvement. In addition, glycoproteins extracted from each plant can be used by mixing two or more kinds.

Test Example 8 Evaluation of Antioxidant Potency (DPPH)

Powder products were prepared by drying Example 1, Example 2, Example 3, Example 4, Example 5, and Example 6, which are the six kinds of glycoprotein liquid products prepared in Example, by using a freeze dryer. . Thereafter, 100 g of its dry weight was prepared by dissolving it in DMSO at a 10% concentration. As a positive control, vitamin C was dissolved in water to make 1%. DPPH (1,1-diphenyl-2-picrylhydrazyl) was prepared to have a concentration of 100 μM in ethanol. After diluting the prepared glycoprotein solution and vitamin C solution by 1/2 step with ethanol to make a solution at five concentrations, 10 μl of each prepared solution and 190 μl of DPPH solution were mixed and incubated at 37 ° C. for 30 minutes. Thereafter, the OD value was measured at 540 nm. On the other hand, the untreated group treated with nothing was tested as a negative control, and the measured value is shown in Figure 1 by evaluating the antioxidant power of each example when the non-treated group to 100. In FIG. 1, the lower the Y-axis value is 100, the greater the antioxidant power.

As can be seen in Figure 1, each example has an excellent antioxidant effect compared to the negative control, the higher the concentration was increased the effect. Therefore, the glycoprotein extracted from each plant was confirmed to have excellent antioxidant effect. And glycoproteins extracted from each plant can be used by mixing two or more kinds.

Hereinafter, the glycoprotein extracted from at least one selected from the group consisting of Bergenia cordifolia, Rhodiola rossia, Komarum parustre, Avis civicica, Betula pendula buzz, and Innotus oblixus Although the formulation examples of the cosmetic composition, the pharmaceutical composition and the health food composition including the composition containing as an active ingredient are described in more detail, the cosmetic composition, the pharmaceutical composition and the health food composition are applicable to various formulations, which It is not intended to be limiting but merely illustrative.

Formulation Example 1 Flexible Cosmetic (Skin Lotion)

According to the composition described in Table 11 below was prepared a flexible cosmetic water in a conventional manner.

Compounding ingredient Content (% by weight) Glycoprotein extracted from each plant 0.1 glycerin 3.0 Butylene glycol 2.0 Propylene glycol 2.0 Carboxy Vinyl Polymer 0.1 Fiji-12 nonylphenyl ether 0.2 Polysorbate 80 0.4 ethanol 10.0 Triethanolamine 0.1 Preservative, coloring, fragrance Quantity Purified water Balance

Formulation Example 2 Nutritious Longevity (Milk Lotion)

Nutrients were prepared in a conventional manner according to the composition shown in Table 12.

Compounding ingredient Content (% by weight) Glycoprotein extracted from each plant 0.1 glycerin 3.0 Butylene glycol 3.0 Propylene glycol 3.0 Carboxy Vinyl Polymer 0.1 Beeswax 4.0 Polysorbate 60 1.5 Caprylic / Capric Triglycerides 5.0 Squalane 5.0 Sorbitassquioleate 1.5 Liquid paraffin 0.5 Cetearyl Alcohol 1.0 Triethanolamine 0.2 Preservative, coloring, fragrance Quantity Purified water Balance

Formulation Example 3 Nutrition Cream

Nutritional cream was prepared in a conventional manner according to the composition described in Table 13.

Compounding ingredient Content (% by weight) Glycoprotein extracted from each plant 0.1 glycerin 3.0 Butylene glycol 3.0 Liquid paraffin 7.0 Beta Glucan 7.0 Carbomer 0.1 Caprylic / Capric Triglycerides 3.0 Squalane 5.0 Cetearyl Glucoside 1.5 Sorbitan stearate 0.4 Polysorbate 60 1.2 Triethanolamine 0.1 Preservative, coloring, fragrance Quantity Purified water Balance

Formulation Example 4 Massage Cream

To prepare a massage cream in a conventional manner according to the composition described in Table 14.

Compounding ingredient Content (% by weight) Glycoprotein extracted from each plant 0.1 glycerin 8.0 Butylene glycol 4.0 Liquid paraffin 45.0 Beta Glucan 7.0 Carbomer 0.1 Caprylic / Capric Triglycerides 3.0 Beeswax 4.0 Cetearyl Glucoside 1.5 Sesqui oleic acid sorbitan 0.9 Vaseline 3.0 paraffin 1.5 Preservative, coloring, fragrance Quantity Purified water Balance

[Formulation Example 5] Pack

To prepare a pack in a conventional manner according to the composition described in Table 15 below.

Compounding ingredient Content (% by weight) Glycoprotein extracted from each plant 0.1 glycerin 4.0 Polyvinyl alcohol 15.0 Hyaluronic acid extract 5.0 Beta Glucan 7.0 Allantoin 0.1 Nonyl Phenyl Ether 0.4 Polysorbate 60 1.2 ethanol 6.0 Preservative, coloring, fragrance Quantity Purified water Balance

[Formulation Example 6] Ointment in the external preparation for skin

The ointment was prepared in a conventional manner according to the composition described in Table 16 below.

Compounding ingredient Content (% by weight) Glycoprotein extracted from each plant 0.1 glycerin 8.0 Butylene glycol 4.0 Liquid paraffin 15.0 Beta Glucan 7.0 Carbomer 0.1 Caprylic / Capric Triglycerides 3.0 Squalane 1.0 Cetearyl Glucoside 1.5 Sorbitan stearate 0.4 Cetearyl Alcohol 1.0 Beeswax 4.0 Preservative, coloring, fragrance Quantity Purified water Balance

Formulation Example 7 Preparation of Powder

Glycoprotein extracted from each plant ............ 100 mg

Lactose ................................... 100 mg

Talc ........................................ 10 mg

Maintenance ...................................... 5 mg

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

Formulation Example 8 Preparation of Tablet

Glycoprotein extracted from each plant ........ 50 mg

Corn starch ..... 100 mg

Lactose ............... 100 mg

Magnesium stearate ........ 2 mg

Vitamin C ............ 50 mg

After mixing the above components, tablets are prepared by tableting according to a conventional method for preparing tablets.

Formulation Example 9 Preparation of Capsule

Glycoprotein extracted from each plant ............... 50 mg

Corn starch ..... 100 mg

Lactose ......................................... 100 mg

Magnesium Stearate ......... 2 mg

Vitamin C ........................ 50 mg

Serine ... mg

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

Formulation Example 10 Preparation of Liquid

Glycoprotein extracted from each plant ............. 100 mg

Isomerized sugar ... ... 10 g

Mannitol ... .... 5 g

Vitamin C ... ... 50 mg

Serine ... ....... 50 mg

maintain................................................. ....... suitable

Purified water................................................. ...... suitable

According to the conventional method for preparing a liquid, each component is added and dissolved in purified water, lemon flavor is added, the above components are mixed, purified water is added, the whole is adjusted to 100 ml by adding purified water, and then filled in a brown bottle. The solution is prepared by sterilization.

Formulation Example 11 Preparation of Health Food

Glycoprotein extracted from each plant .................... 1000 mg

Vitamin mixtures

Vitamin A Acetate ............... 70 μg

Vitamin E ......................................... 1.0 mg

Vitamin B1 .................................. 0.13 mg

Vitamin B2 ................................... 0.15 mg

Vitamin B6 ......................................... 0.5 mg

Vitamin B12 ......................... 0.2 μg

Vitamin C ......................................... 10 mg

Biotin ......................................... 10 μg

Nicotinic Acid Amide ... 1.7 mg

Folic acid ......................................... 50 ㎍

Calcium Pantothenate ......................................... 0.5 mg

Mineral mixture

Ferrous Sulfate ............... 1.75 mg

Zinc Oxide ............... 0.82 mg

Magnesium Carbonate ... 25.3 mg

Potassium monophosphate ......................................... 15 mg

Dibasic Calcium Phosphate ......................................... 55 mg

Potassium Citrate ... 90 mg

Calcium Carbonate ... 100 mg

Magnesium Chloride ......................................... 24.8 mg

The composition ratio of the above-mentioned vitamin and mineral mixtures is composed of relatively suitable ingredients suitable for health foods in a preferred formulation example, but the formulation ratio may be arbitrarily modified. The granules may be prepared and used for preparing a health food composition according to a conventional method.

Formulation Example 12 Preparation of Healthy Drink

Glycoprotein extracted from each plant ..................... 1000 mg

Citric Acid ... ......... 1000 mg

oligosaccharide................................................. ........ 100 g

Plum concentrate ..................... ......... 2 g

Taurine ... ............. 1 g

Purified water is added to the whole ........... 900 ㎖

After mixing the above components according to a conventional healthy beverage production method, and then stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained by sterilization in a sterilized 2 L container, sealed sterilization and then refrigerated and stored in the present invention For the preparation of healthy beverage compositions.

Although the composition ratio is composed of relatively suitable components for the preferred beverage in the preferred formulation example, the composition ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and use purpose.

Figure 1 is a graph of the results of Test Example 8 measuring the antioxidant power of glycoprotein and vitamin C extracted from each of six plants using DPPH.

Claims (10)

Bergenia cordifolia ), Rhodiola rosea ), Comarum palustre ), Abies Sibirica Sibirica , Betula pendula Buds ), and Inonotus Obliquus ( Inonotus Obliquus ) A skin wrinkle preventing or improving composition containing as an active ingredient glycoprotein extracted from at least one selected from the group consisting of. Contains as an active ingredient glycoproteins extracted from one or more selected from the group consisting of bergenia cordifolia, rhodiola roscia, comarum parustre, abyss civicica, betula pendula buzz, and inototus oblicus Skin whitening composition. Contains as an active ingredient glycoproteins extracted from one or more selected from the group consisting of bergenia cordifolia, rhodiola roscia, comarum parustre, abyss civicica, betula pendula buzz, and inototus oblicus Inflammation preventing or ameliorating composition. Contains as an active ingredient glycoproteins extracted from one or more selected from the group consisting of bergenia cordifolia, rhodiola roscia, comarum parustre, abyss civicica, betula pendula buzz, and inototus oblicus Skin moisturizing composition. Contains as an active ingredient glycoproteins extracted from one or more selected from the group consisting of bergenia cordifolia, rhodiola roscia, comarum parustre, abyss civicica, betula pendula buzz, and inototus oblicus Skin regeneration promoting composition. Contains as an active ingredient glycoproteins extracted from one or more selected from the group consisting of bergenia cordifolia, rhodiola roscia, comarum parustre, abyss civicica, betula pendula buzz, and inototus oblicus Scar prevention or improvement composition. Contains as an active ingredient glycoproteins extracted from one or more selected from the group consisting of bergenia cordifolia, rhodiola roscia, comarum parustre, abyss civicica, betula pendula buzz, and inototus oblicus Atopic Skin Improvement Composition. Contains as an active ingredient glycoproteins extracted from one or more selected from the group consisting of bergenia cordifolia, rhodiola roscia, comarum parustre, abyss civicica, betula pendula buzz, and inototus oblicus Antioxidant enhancing composition. Contains as an active ingredient glycoproteins extracted from one or more selected from the group consisting of bergenia cordifolia, rhodiola roscia, comarum parustre, abyss vicica, betula pendula buzz, and inototus oblicus Anti-aging or improving composition. The method according to any one of claims 1 to 9, Glycoproteins extracted from one or more selected from the group consisting of bergenia cordifolia, rhodiola roscia, comarum parustre, abyss civicica, betula pendula buzz, and innotus olivus A composition containing 0.1 to 30% by weight.

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