KR20210097239A - Anti-aging composition comprising functional peptides, amino acids, callus extract and fermented materials - Google Patents

Abstract

Disclosed is an anti-aging composition for effectively preventing or alleviating skin aging and wrinkles and having an excellent antioxidant effect. The present invention provides an anti-aging composition comprising acetyl hexapeptide-8, hexapeptide-9, tripeptide-1, acetyl tetrapeptide-5, hexapeptide-11, Leontopodium alpinum callus culture extract, Bifida ferment lysate, Isoleucine, valine, leucine, arginine, lysine, phenylalanine, methionine, histidine, tryptophan and threonine.

Description

Anti-aging composition comprising functional peptides, amino acids, callus extract and fermented product {Anti-aging composition comprising functional peptides, amino acids, callus extract and fermented materials}

The present invention relates to an anti-aging composition, and more particularly, to an anti-aging composition that promotes skin cell regeneration, improves skin wrinkles, and exhibits antioxidant effects.

Among the theories of human aging, the most popular theory is the aging caused by free radicals. More than 95% of the oxygen consumed by the human body is reduced to water by combining with electrons generated in the metabolic process of cells, but 2 to 3% of oxygen is incompletely reduced to absorb electrons in the free radical process. It causes destructive action, which is called active oxygen. When reactive oxygen species act on the cell membrane composed of lipids and proteins, lipid peroxidation is induced and the content of the final product, Malondialdehyde (MDA), is increased. MDA chemically modifies low-density lipoprotein (LDL) in the lining of blood vessel walls, and the modified low-density lipoprotein newly synthesizes cholesterol in macrophages to form foam cells as esters are deposited. It is known that it causes diseases such as arteriosclerosis, liver disease, and various cancers by lowering this physiological function, which eventually leads to aging and genetic disorders. With aging, the concentration of active oxygen (O ₂ ^- , OH ^- ), nitric oxide (NO), and hydrogen peroxide (H ₂ O ₂ ) in the living body increases, which promotes various diseases and cell aging. .

In particular, free radicals cause damage to skin cells. If there are too many free radicals, the antioxidant defense system of the body is broken down, and as a result, cellular components such as proteins, lipids, and DNA are damaged, and cellular functions are altered, ultimately promoting skin aging. In addition, free radicals damage the connective tissue. Free radicals act on collagen metabolism to directly destroy collagen and cause collagen deficiency.

Free radicals occur in the process of breathing and digestion, which are essential to sustain life, as well as various factors that threaten our health, such as smoking, stress, excessive exercise, skin diseases, viral infections, and ultraviolet rays. Therefore, humans are continuously exposed to the threat of free radicals, and therefore our body has various antioxidant mechanisms in response to the damage caused by free radicals to protect cells in various ways. Factors involved in the antioxidant power of inhibiting free radicals and oxidizing substances in the human body can be divided into two main categories: the antioxidant enzyme system and the non-enzymatic antioxidant. Among the antioxidant enzyme systems in the body, the SOD (superoxide dismutase) enzyme that is _{generated from copper, zinc, or manganese and removes O 2} ^{- is representative.} It _{removes H 2} O ₂ , and GPX is known to repair and restore damaged cells to their original state. Non-enzymatic antioxidants are divided into those that are made in the body and those that must be supplied from outside the body. Antioxidants in the body include albumin, ferritin, transferrin, glutathione (GSH), uric acid, bilirubin, and melatonin, which are metal-binding proteins. As an antioxidant of interest, there is CoQ10 coenzyme, which is present in mitochondria and plays an important role in energy metabolism and antioxidant action.

Skin aging is a natural phenomenon that progresses with the passage of time, and like human aging, there are individual differences, but it is not a property that can be fundamentally prevented. However, the causes of skin aging are not only intrinsic factors that progress over time, but also extrinsic factors that can be prevented, such as smoking, excessive drinking, malnutrition, and chronic exposure to sunlight. Therefore, it is known that skin aging and skin wrinkles derived from it cannot be prevented for a lifetime, but can be delayed. It can be said that this is an area in which research is focused.

In the cosmetic technology field, many efforts are being made to overcome wrinkles, pigmentation, loss of elasticity, skin dryness, hair loss, and lack of shine due to aging-related skin changes. The skin undergoes various changes through aging. First, the thickness of the epidermis, dermis, and subcutaneous tissue, which are components of the skin, becomes thinner and the extracellular matrix (ECM) component that gives elasticity to the skin changes. With age, its production decreases rapidly and has a close relationship with the generation of wrinkles, and collagen, elastin, proteoglycans, and glucosaminoglycans that make up the skin connective tissue ), laminin, fibronectin, etc. are oxidized and lose their functions, so that the skin loses elasticity and wrinkles are excessively formed, changing to senile skin.

The connective tissue fibers of the extracellular matrix include glue fibers (collagen fibers), reticular fibers, and elastic fibers (elastic fibers), of which collagen accounts for about 70% of the skin connective tissue. Most are formed in fibroblasts of the skin. Collagen content in the skin connective tissue decreases with age, which is due to a decrease in collagen synthesis and promotion of decomposition. Therefore, the decrease in collagen biosynthesis and the promotion of collagen decomposition are the biggest causes of skin wrinkles.

The collagen biosynthesis process is regulated by many factors involved in the transcription level and post-translation level, and in the case of collagen degradation, collagen is decomposed by ultraviolet rays. Collagen decomposition is promoted by promoting the expression of matrix metalloproteases (MMP), such as genase (Collagenase), and the collagen content is reduced. In addition, as the transformation of collagen is accelerated by the external environment, the skin becomes wrinkled and deep. As a result, skin aging is caused by non-homogeneity of cells, loss of elastin, destruction of collagen, reduction and delay of collagen synthesis, and the like. Therefore, although skin aging occurs in the epidermis, it can be seen as a phenomenon that occurs in the dermis rather than this.

Various cosmetic compositions are being studied to solve the problems of skin aging, and the effect of improving skin wrinkles is showing visible results in some cases. For example, clinical results of skin wrinkle removal for retinoids, which are widely used to improve skin wrinkles, blemishes, and pigmentation, among others, retinol have been reported variously, and cosmetics containing retinol are used for skin wrinkles formed by sunlight. It effectively improved skin sagging and loss of elasticity. Retinol is a raw material whose effectiveness has been verified enough to be registered as a raw material for wrinkle function, but it has been constantly raised about its unique skin irritation and stability issues. there is a problem.

In order to effectively prevent or improve skin aging and wrinkles, it is necessary to develop a new antioxidant and anti-aging composition using raw materials whose antioxidant efficacy has been confirmed and raw materials whose effects on skin aging have been accurately verified. It is important to clarify with

Accordingly, the present inventors have made diligent efforts to develop a composition that can effectively prevent or improve skin aging and wrinkles, acetyl hexapeptide-8 (Acetyl Hexapeptide-8), hexapeptide-9 (Hexapeptide-9), Tripeptide-1, Acetyl Tetrapeptide-5, Hexapeptide-11, Leontopodium alpinum callus culture extract, Bifida fermentation lysate ( Bifida Ferment Lysate, Isoleucine, Valine, Leucine, Arginine, Lysine, Phenylalanine, Methionine, Histidine, Tryptophan) and It was found that the composition containing threonine exhibits a synergistic effect between the components, thereby exhibiting a very excellent antioxidant effect, wrinkle improvement effect, and skin regeneration effect, and completed the present invention.

Accordingly, an object of the present invention is to provide an anti-aging composition that effectively prevents or improves skin aging and wrinkles, and has an excellent antioxidant effect.

Another object of the present invention is to provide a cosmetic composition, a pharmaceutical composition and a functional food composition comprising the anti-aging composition.

In order to achieve the above object, the present invention provides acetyl hexapeptide-8 (Acetyl Hexapeptide-8), hexapeptide-9 (Hexapeptide-9), tripeptide-1 (Tripeptide-1), acetyl tetrapeptide-5 (Acetyl Tetrapeptide) -5), hexapeptide -11 (Hexapeptide-11), Edelweiss callus culture extract (Leontopodium alpinum extract callus culture), bipyridinium the lysate (Bifida Ferment lysate), isoleucine (isoleucine) for fermentation, Val (valine), leucine (leucine ), arginine, lysine, phenylalanine, methionine, histidine, tryptophan, and an anti-aging composition comprising Threonine, and a cosmetic composition comprising the same; A pharmaceutical composition and a functional food composition are provided.

Hereinafter, the present invention will be described in detail.

Each of the functional raw materials constituting the composition of the present invention acts on the skin through different mechanisms, and helps to prevent skin aging by promoting antioxidant, collagen production and epidermal cell regeneration. As the raw materials exhibit a synergistic effect with each other, the effect is significantly increased compared to using them alone.

According to one embodiment of the present invention, acetyl hexapeptide-8 (Acetyl Hexapeptide-8), hexapeptide-9 (Hexapeptide-9), tripeptide-1 (Tripeptide-1), acetyl tetrapeptide-5 (Acetyl Tetrapeptide- 5), Hexapeptide-11, Edelweiss callus culture extract, Bifida Ferment Lysate, Isoleucine, Valine, Leucine , Arginine, Lysine, Phenylalanine, Methionine, Histidine, Tryptophan and Threonine The composition of the present invention containing all of the 17 components It was found that it can be used in a composition for preventing aging of the skin through a significantly superior antioxidant effect, collagen production promotion, and collagen degrading enzyme expression inhibitory effect compared to a composition that does not contain any one of them.

That is, the composition of the present invention was confirmed to be very excellent in the effect of inhibiting the activity of collagen-degrading enzymes such as collagenase and MMP-1 (Matrix Metalloproteinase-1), and the effect of promoting intracellular collagen production and It was confirmed that the effect of improving the regenerative ability was also very excellent, and it was found that it can be very usefully utilized in the development of an anti-aging functional composition.

According to another embodiment of the present invention, the composition of the present invention does not show toxicity to human-derived cells and thus has very high utility as an anti-aging functional composition. According to the present invention, as a result of treating the composition in HaCat cells and NIH3T3 cells at a concentration of 100 ng/ml to 1,000 μg/ml, cytotoxicity was not measured, and a large change was observed even when the morphology of the cells was visually confirmed. not observed.

According to another embodiment of the present invention, it was confirmed that the composition of the present invention exhibited excellent thermal stability even at a temperature of 50° C., and was stable even under sunlight conditions. That is, it means that the composition of the present invention can be advantageously applied to products requiring long-term storage, such as pharmaceuticals, quasi-drugs, and cosmetics.

In the present invention, the 'skin regeneration effect' refers to the recovery of skin tissue against damage caused by external and internal causes of the skin. The damage caused by the external cause may include ultraviolet rays, external pollutants, wounds, trauma, and the like, and the damage caused by the internal cause may include stress.

In the present invention, the 'wrinkle improvement effect' refers to suppressing or inhibiting the generation of wrinkles on the skin, or alleviating the wrinkles already generated.

In the present invention, the term 'antioxidation' refers to inhibition of cell oxidation by highly reactive free radicals or reactive oxygen species (ROS) depending on intracellular metabolism or oxidative stress caused by the influence of ultraviolet rays. This includes reducing the damage to cells by removing free radicals or reactive oxygen species.

In the present invention, the acetyl hexapeptide-8 is a substance also known as Argireline or acetyl hexapeptide-3, and is known to have a very excellent anti-wrinkle effect. In particular, acetyl hexapeptide-8 exhibits an effect on wrinkles by inducing muscle relaxation with a mechanism similar to Botox. The acetylhexapeptide-8 may be included in an amount of 0.001 to 1.0% (w/v), preferably 0.005 to 0.5% (w/v), most preferably 0.01 to 0.02% ( w/v) may be included.

In the present invention, the hexapeptide-9 is a peptide consisting of the amino acid sequence of Gly-Pro-Gln-Gly-Pro-Gln, and is an analog of Human Collagen IV and XVII, which are two basic types of membrane collagen. It heals damaged cells and induces skin regeneration, confirming the efficacy of skin wrinkle and anti-aging. The hexapeptide-9 may be included in an amount of 0.001 to 1.0% (w/v) relative to the total composition of the present invention, preferably 0.005 to 0.5% (w/v), more preferably 0.01 to 0.05% (w) /v), most preferably 0.01 to 0.02% (w/v) content.

In the present invention, the tripeptide-1 is a peptide consisting of the amino acid sequence of Gly-His-Lys, and it helps to regenerate the skin by removing the damage to collagen and elastin caused by skin damage and suppressing scar formation and promoting the generation of stem cells. In particular, it is effective for cell regeneration by promoting the production of growth factors (EGF, VEGF). The hexapeptide-1 relative to the total composition of the present invention may be included in an amount of 0.001 to 1.0% (w/v), preferably 0.005 to 0.5% (w/v), more preferably 0.01 to 0.05% (w) /v), most preferably 0.01 to 0.02% (w/v) content.

In the present invention, the acetyl tetrapeptide-5 is a peptide consisting of the amino acid sequence of Ac-beta-Ala-His-Ser-His. It is a peptide material with excellent anti-aging effect. The hexapeptide-5 may be included in an amount of 0.001 to 1.0% (w/v) relative to the total composition of the present invention, preferably 0.005 to 0.5% (w/v), more preferably 0.01 to 0.05% (w) /v), most preferably 0.01 to 0.02% (w/v) content.

In the present invention, the hexapeptide-11 is a peptide consisting of the amino acid sequence of Phe-Val-Ala-Pro-Phe-Pro. The sequence was first identified in brewer's yeast, and it inhibits the aging process of fibroblasts to make the skin young. It is known to have anti-aging effects that keep it healthy. The hexapeptide-11 relative to the total composition of the present invention may be included in an amount of 0.001 to 1.0% (w/v), preferably 0.005 to 0.5% (w/v), more preferably 0.01 to 0.05% (w) /v), most preferably 0.01 to 0.02% (w/v) content.

In the present invention, the edelweiss callus culture extract refers to a material made by inducing a callus from the stem of edelweiss and extracting the prepared callus. In the composition of the present invention, the edelweiss callus culture extract may be included in an amount of 0.5 to 20% (v/v) relative to the total composition, preferably in an amount of 1 to 15% (v/v).

In the present invention, the preparation of the edelweiss callus culture extract is not particularly limited, but preferably (a) after disinfecting and peeling off the epidermis of the stem of edelweiss in order to prevent or improve skin aging and wrinkles of the final composition, and to implement an antioxidant effect Separating the cambium from the internal stem tissue, inoculating the edelweiss tissue into a medium to induce callus and then proliferating; (b) inoculating the induced callus in a bioreactor and culturing; and (c) drying the cultured callus and then extracting by adding a solvent; and can be prepared, specifically, by the following method:

(a) edelweiss callus and callus induction by tissue culture

After sterilizing and sterilizing the stem of edelweiss, cut it to a thickness of about 1 to 5 mm to remove the epidermis, and then separate the cambium from the internal tissue of the stem. The isolated edelweiss tissue is inoculated in Modified MS medium to induce callus. Proliferating the induced callus tissue in Modified MS medium, and maintaining and using the callus tissue while subcultured at intervals of 1 to 3 weeks;

(b) mass production of callus

With respect to the edelweiss callus induced in step (a), in the same liquid medium used for callus induction, a growth regulator, etc. was added to cut the callus derivative into 5 to 10 mm size, inoculated into a bioreactor, and inoculated at 20 to 25 ° C. mass culturing at a temperature and an air injection rate of 100 to 1,000 cc/min; and

(c) preparation of callus extract

When the callus culture is completed, the callus is dried by hot air drying, and purified water 7 to 10 times the volume of the dried callus is added, extracted at 100 to 121° C. for 10 to 100 minutes, and then extracted at room temperature for 12 to 48 hours to complete the extraction.

In the present invention, the Bifida fermentation lysate refers to a material prepared by culturing a microorganism of the genus Bifidobacterium, which is a kind of probiotic lactic acid bacteria, and then dissolving the culture solution. In the composition of the present invention, the Bifida fermentation lysate may be included in an amount of 0.5 to 20% (v/v), preferably 1 to 15% (v/v) relative to the total composition.

In the present invention, the preparation of the fermented Bifida lysate is not particularly limited, but is preferably inoculated with a microorganism of the genus Bifidobacterium to prevent or improve skin aging and wrinkles of the final composition, and to implement an antioxidant effect, followed by culturing and fermentation. can be prepared, specifically B. actinocoloniiforme, B. adolescentis, B. angulatum, B. animalis, B. aquikefiri, B. asteroides, B. biavatii, B.bifidum, B. bohemicum, B. bombi, B. boum , B. breve, B. callitrichos, B. catenulatum, B. choerinum, B. commune, B. coryneforme, B. cuniculi, B. crudilactis, B. denticolens, B. dentium, B. eulemuris, B. faecale, B Gallicum, B. gallinarum, B. hapali, B. indicum, B. inopinatum, B. kashiwanohense, B. lemurum, B. longum, B. magnum, B. merycicum, B. minimum, B. mongoliense, B. moukalabense , B. myosotis, B. pseudocatenulatum, B. pseudolongum, B. psychraerophilum, B. pullorum, B. reuteri, B. ruminantium, B. saguini, B. scardovii, B. stellenboschense, B. stercoris, B. saeculare, B. subtile, B. thermacidophilum, B. thermophilum,,B. tissieri, B. infantis, and B. tsurumiense can be prepared by inoculating a microorganism of the genus Bifidobacterium selected from the group consisting of, culturing and fermenting.

More specifically, the culture solution of one selected strain ^{is adjusted to 10 7} to 10 ¹⁰ cfu/ml and inoculated, and may be prepared by fermentation at 25 to 45° C. for 2 to 10 days. The Bifidobacterium strain in the present invention preferably be a Bifidobacterium ronggeom (B. longum), Bifidobacterium bipyridinium bushes (B. bifidum) or Bifidobacterium inpen tooth (B. infantis) However, the present invention is not limited thereto.

In the present invention, the isoleucine, valine, leucine, arginine, lysine, phenylalanine, methionine, histidine, tryptophan and threonine (Threonine) (hereinafter referred to as 'amino acid mixture') is an essential amino acid that cannot be synthesized by itself among 20 kinds of amino acids. Amino acids are a component of peptides and proteins. They have a wide range of benefits in skin care, and amino acids are naturally present in the skin as part of what are called natural moisturizing factors (NMFs), which exert antioxidant effects by causing them to produce more antioxidants. In addition, amino acids strengthen the skin's defense system and help with aging symptoms caused by environmental damage. In the present invention, the amino acid mixture may be included in an amount of 0.05 to 10% (w/v) relative to the total composition, preferably 0.5 to 5% (w/v), most preferably 0.1 to 2% (w/v) ) may be included.

In another aspect, the present invention discloses a cosmetic composition, a pharmaceutical composition and a functional food composition comprising the above 17 components.

In the present invention, the cosmetic composition may be applied in a gel type, skin type, cream type, ointment type, etc., but is not limited thereto. The above composition can be suitably prepared by a known method by adding an appropriate conventional softening agent, emulsifying agent, thickening agent or other substances known in the art according to its type.

The gel-type composition may be prepared by adding an emollient such as trimethylolpropane, polyethylene glycol, or glycerin, a solvent such as propylene glycol, ethanol, or isostatic alcohol, and purified alcohol.

The skin type composition includes fatty alcohols such as stearyl alcohol, myristyl alcohol, behenyl alcohol, arachidyl alcohol, isostearyl alcohol, isocetyl alcohol, butylene glycol, glycerin, allantoin, methyl paraben, EDTA- It can be prepared by adding 2-sodium, xanthan gum, dimethicone, polyethylene glycol-60 hydrogenated castol oil, polysorbate 60 and purified water.

The cream-type composition comprises fatty alcohols such as stearyl alcohol, myristyl alcohol, behenyl alcohol, arachidyl alcohol, isostearyl alcohol, isocetyl alcohol, lecithin, phosphatidylcholine, phosphatidylethanolamine, sofatizylserine, sofphatidyl. Lipids such as inositol; It can be prepared by adding a solvent such as propylene glycol and purified water.

The ointment-type composition may be prepared by adding a softening agent, an emulsifying agent, and waxes such as microcrystalline lead, paraffin, ceresin, beeswax, beeswax, and vaseline.

The cosmetic composition of the present invention may additionally contain one or more ingredients exhibiting the same or similar function in addition to the active ingredient. Non-limiting examples thereof may be any one or more selected from the group consisting of vitamin C, retinoic acid, TGF, animal placental-derived protein, betulinic acid, and chlorella extract, but is not limited thereto. In addition, the cosmetic composition of the present invention may further contain excipients including fluorescent substances, fungicides, hydrotropes, humectants, fragrances, fragrance carriers, proteins, solubilizers, sugar derivatives, sunscreens, vitamins, plant extracts, etc. can

In the present invention, the functional food composition is added to food materials such as beverages, teas, spices, gum, and confectionery, or is a food prepared in encapsulation, powdering, suspension, etc. However, unlike general drugs, it has the advantage that there are no side effects that may occur when taking the drug for a long period of time by using food as a raw material.

Since the functional food composition of the present invention obtained in this way can be ingested on a daily basis, it is very useful because high skin regeneration, anti-wrinkle and antioxidant effects can be expected.

When the composition is used as a food additive, the composition may be added as it is or used together with other foods or food ingredients, and may be appropriately used according to a conventional method. The mixed amount of the active ingredient may be suitably determined according to the purpose of its use (prevention, health or therapeutic treatment). In general, in the production of food or beverage, the composition of the present invention is added in an amount of 15 wt$ or less, preferably 10 wt% or less, based on the total weight of the raw material. However, in the case of long-term intake for health and hygiene or health control, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range. .

There is no particular limitation on the type of the food. Examples of foods to which the above substances can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, tea, drinks, There are alcoholic beverages and vitamin complexes, and all functional foods in the ordinary sense are included.

The functional food composition of the present invention may be prepared as a functional beverage by containing various flavoring agents or natural carbohydrates as additional ingredients, such as conventional beverages. The above-mentioned natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. As the sweetener, natural sweeteners such as taumatine and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like can be used. The proportion of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 ml of the composition of the present invention.

In addition to the above, the functional food of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol , a carbonation agent used in carbonated beverages, and the like. In addition, the health food of the present invention may contain fruit for the production of natural fruit juice, fruit juice beverage, and vegetable beverage. These components may be used independently or in combination. The proportion of these additives is not particularly important, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

In the present invention, the pharmaceutical composition may include a pharmaceutically acceptable carrier in addition to the active ingredient, and these carriers are commonly used in formulations, such as lactose, dextrose, sucrose, sorbitol, mannitol, starch, Gum acacia, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, stearic acid Magnesium and mineral oil, and the like, but are not limited thereto. The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, and the like, in addition to the above components. Suitable pharmaceutically acceptable carriers and agents are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

A suitable dosage of the pharmaceutical composition according to the present invention may vary depending on factors such as formulation method, administration method, age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and response sensitivity of the patient. can be prescribed. Meanwhile, the dosage of the pharmaceutical composition of the present invention is preferably 0.0001 to 100 mg/kg (body weight) per day. 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 topical application to the skin, intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, etc. . Considering that the pharmaceutical composition of the present invention exhibits the effects of antioxidant, wrinkle improvement and skin regeneration, it is preferable to be administered orally or applied topically to the skin.

The concentration of the active ingredient contained in the composition of the present invention may be determined in consideration of the therapeutic purpose, the patient's condition, the required period, etc., and is not limited to a concentration within a specific range.

The pharmaceutical composition of the present invention is prepared in unit dosage form by formulating using a pharmaceutically acceptable carrier or excipient according to a method that can be easily carried out by a person of ordinary skill in the art to which the present invention pertains, or It can be prepared by pouring into a multi-dose container. In this case, the formulation may be prepared as any one formulation selected from injections, creams, patches, sprays, ointments, warning agents, lotions, liniments, pastas, and cataplasmas. When the composition is used as an external preparation for skin, in addition, a fatty substance, an organic solvent, a solubilizer, a thickening agent and a gelling agent, an emollient, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance, a surfactant, water, Ionic or nonionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic actives, lipid vesicles or external preparations for skin It may contain adjuvants commonly used in the field of dermatology, such as any other ingredients. In addition, the above ingredients may be introduced in an amount generally used in the field of dermatology.

The anti-aging composition according to the present invention prevents aging of the skin through the anti-oxidation effect, the wrinkle improvement effect and the cell regeneration effect due to the promotion of collagen production and decomposition inhibition, so that the skin can be kept healthy and young, so that it can be used in the manufacture of functional foods, cosmetics and pharmaceuticals. It can be very useful.

1 is a graph confirming the MMP-1 inhibitory ability of the composition according to the present invention in Example 4;
2 is a graph confirming the intracellular collagen production promoting effect of the composition according to the present invention in Example 5;
3 is a photograph confirming the skin cell regeneration promoting effect of the composition according to the present invention in Example 6;
4 and 5 are graphs showing the results of measuring the thermal stability and photostability of the anti-aging cosmetic composition according to the present invention in Example 7;
6 and 7 are graphs confirming the cytotoxicity of the anti-aging cosmetic composition according to the present invention in Example 7 through MTT assay in HDF cells and HACAT cells.

Hereinafter, the present invention will be described in detail through preferred embodiments. Prior to this, the terms or words used in the present specification and claims should not be construed as being limited to their ordinary or dictionary meanings, and the inventor should properly understand the concept of the term in order to best describe his invention. Based on the principle that can be defined, it should be interpreted as meaning and concept consistent with the technical idea of the present invention. Therefore, since the configuration of the embodiments described in the present specification is only the most preferred embodiment of the present invention and does not represent all the technical spirit of the present invention, various equivalents and modifications that can be substituted for them at the time of the present application It should be understood that there may be

<Example 1: Preparation of anti-aging composition>

Functional peptide, Edelweiss callus culture extract, Bifida fermentation lysate and 10 essential amino acids (Isoleucine, Valine, Leucine, Arginine, Lysine, Phenylalanine, Methionine) (Methionine), histidine (Histidine), tryptophan (Tryptophan) and threonine (Threonine)) was prepared by combining the anti-aging composition.

As functional peptides, Acetyl Hexapeptide-8, Hexapeptide-9, Tripeptide-1, Acetyl Tetrapeptide-5 and Hexapeptide -11 (Hexapeptide-11) was obtained through a known solid phase synthesis method, and in the case of Bifida fermentation lysate, the culture medium of the Bifidobacterium longum ^{strain was adjusted to 10 9} cfu/ml and then at 37 ° C. It was prepared by fermentation for 10 days. After sterilizing and sterilizing the Edelweiss callus culture extract, the stem of Edelweiss is cut to a thickness of 1 to 5 mm, the epidermis is peeled off, and the cambium is separated from the stem internal tissue, and the separated Edelweiss tissue is inoculated into Modified MS medium to induce callus Then, the induced callus tissue is grown in Modified MS medium and subcultured at 2-week intervals to maintain the callus tissue, and when the callus culture is completed, the callus is dried by hot air drying, and then 7 to 10 times the volume of the dry callus. of purified water was added and extracted at 100 to 121 °C for 10 to 30 minutes, followed by extraction at room temperature for 24 hours to obtain an extract.

A total of 18 combinations were selected by controlling the concentrations of the prepared functional peptides, Edelweiss callus culture extract and Bifida fermentation lysate, and 10 essential amino acids, and the composition ratio of the components in the selected combination is shown in Table 1 below.

Acetyl Hexapeptide-8, Hexapeptide-9, Tripeptide-1, Acetyl Tetrapeptide-5, Hexapeptide-11 -11) and 10 essential amino acids were dissolved in water at the concentrations shown in Table 1 below (w/v), and the Edelweiss callus culture extract and Bifida fermentation lysate were prepared according to the above method, and then the concentrations shown in Table 1 below. was added to water (v/v) to prepare a composition. Amino acid mix (Mix) was dissolved so that the total weight of the final mixture by mixing all 10 amino acids in a content of 0.1% by weight is 1.0% (w / v).

division Acetyl Hexapeptide-8
(w/v%) Hexapeptide-11
(w/v%) Hexapeptide-9
(w/v%) Acetyl Tetrapeptide-5
(w/v%) Tripeptide-1
(w/v%) Amino Acid Mix
(w/v%) Bifida Ferment Lysate
(v/v%) Edelweiss callus culture extract
(v/v%) Combination 1 0 0.01 0.01 0.01 0.01 1.0 10 10 Combination 2 0.01 0 0.01 0.01 0.01 1.0 10 10 combination 3 0.01 0.01 0 0.01 0.01 1.0 10 10 combination 4 0.01 0.01 0.01 0 0.01 1.0 10 10 combination 5 0.01 0.01 0.01 0.01 0 1.0 10 10 combination 6 0.01 0.01 0.01 0.01 0.01 0 10 10 combination 7 0.01 0.01 0.01 0.01 0.01 1.0 0 10 combination 8 0.01 0.01 0.01 0.01 0.01 1.0 10 0 combination 9 0.01 0.005 0.005 0.005 0.005 1.0 10 10 combination 10 0 0.005 0.005 0.005 0.005 1.0 10 10 combination 11 0.005 0 0.005 0.005 0.005 1.0 10 10 combination 12 0.005 0.005 0 0.05 0.005 1.0 10 10 combination 13 0.005 0.005 0.005 0 0.005 1.0 10 10 combination 14 0.005 0.005 0.005 0.005 0 1.0 10 10 combination 15 0.005 0.005 0.005 0.005 0.005 0 10 10 combination 16 0.005 0.005 0.005 0.005 0.005 1.0 0 10 combination 17 0.005 0.005 0.005 0.005 0.005 1.0 10 0 combination 18 0.005 0.005 0.005 0.005 0.005 1.0 10 10

<Example 2: Confirmation of antioxidant activity effect>

In order to confirm the antioxidant effect of the composition prepared in Example 1, DPPH scavenging ability and Hydroxy Radical scavenging ability tests were performed according to the following method, and the results are shown in Table 3 below.

[DPPH scavenging ability test method]

(1) Prepare a stock solution according to the concentration of the sample (test substance). In this case, if the solubility is not good, use DMSO. The sample is prepared by mixing with ethanol so that the concentration of the composition shown in Table 1 is 5% (v/v). As a positive control, EGCG (Epigallocatechin gallate) at the same concentration was prepared.

(2) Prepare 100 ml by dissolving 5.9 mg of DPPH (1,1-diphenyl-2-picrylhydrazyl) (Sigma D9132), an indicator to be used in the test, in ethanol.

(3) Prepare a new clean 96-well plate and put 100 μl of the DPPH solution into each well.

(4) Add 90 μl of ethanol to each well of a 96-well plate. Process 10 μl of the prepared sample.

(5) The reaction is stirred at 25° C. for 30 minutes.

(6) Measure the O.D value at 517 nm using a UV spectrophotomer, and calculate the DPPH scavenging ability according to Equation 1 below.

[Equation 1]

[Hydroxy radical scavenging ability test method]

(1) Prepare 0.2 M sodium phosphate monobasic and dibasic, mix 47.5 ㎖ of monobasic and 202.5 ㎖ of dibasic, adjust to 500 ㎖ with purified water, adjust pH to 7.4 to prepare 100 mM phosphate buffer, prepared phosphate buffer Prepared by dissolving 2-deoxy-D-ribose (sigma D5899) to 10 mM, 0.5 mM iron(III) chloride hexahydrate (aldrich 236489) and 10 mM hydrogen peroxide (sigma H1009) in buffer. 10 mM ascorbic acid (aldrich A92902) and 2.8% TCA (Trichloroacetic acid) (sigma T9159) were prepared by dissolving in an aqueous solution, and 1% TBA (4,6-dihydroxy-2-mercaptopyrimidine) (alfa aesar A12681) was prepared by dissolving 0.05 N NaOH It is prepared by dissolving in an aqueous solution.

(2) Prepare the sample (test substance) by mixing it with ethanol so that the concentration of the mixture shown in Table 1 is 5% (v/v).

(3) With reference to Table 2 below, each solution prepared above is dispensed into a vial.

division experimental group control 100 mM phosphate buffer 190 μl 340 μl 10 mM 2-deoxy-D-ribose 100 μl 100 μl 0.5 mM Iron(III) chloride 100 μl - Sample (control: ethanol) 10 μl 10 μl mix 10 mM H ₂ O ₂ 50 μl - 10 mM ascorbic acid 50 μl - ddH ₂ O - 50 μl mix

(4) After incubation at 37° C. for 1 hour, 500 μl/vial of 2.8% TCA is added to terminate the reaction.

(5) After adding 500 μl/vial of 1% TBS, heat at 100° C. for 15 minutes to develop color.

(6) Immerse in ice water for 5 minutes to cool.

(7) After centrifugation at 5,000 rpm for 5 minutes, the supernatant is dispensed in 250 μl/96 well plate.

(8) Measure the absorbance at 532 nm, and calculate the hydroxyl radical scavenging ability according to Equation 2 below.

[Equation 2]

division DPPH scavenging ability (%) Hydroxy radical scavenging ability (%) Control group (untreated group) 0 0 Combination 1 54.7 67.5 Combination 2 53.7 63.7 combination 3 60.4 69.9 combination 4 38.1 49.4 combination 5 57.4 67.3 combination 6 44.5 56.2 combination 7 51.3 60.4 combination 8 50.7 59.9 combination 9 64.4 78.3 combination 10 44.2 58.7 combination 11 46.4 59.1 combination 12 41.4 80.8 combination 13 35.7 46.4 combination 14 43.7 54.5 combination 15 24.1 36.7 combination 16 44.3 56.7 combination 17 46.6 55.2 combination 18 54.8 66.1 positive control (EGCG) 69.4 81.5

Referring to Table 3, DPPH scavenging activity (%) and hydroxyl radical scavenging activity (%) can be confirmed in all combinations, in particular, according to the present invention, acetyl hexapeptide-8 (Acetyl Hexapeptide-8), hexapeptide-9 (Hexapeptide) the -9), tripeptide -1 (tripeptide-1), tetra-acetyl peptide -5 (acetyl Tetrapeptide-5), hexapeptide -11 (hexapeptide-11), Edelweiss callus culture extract (Leontopodium alpinum extract callus culture), bipyridinium When Bifida Ferment Lysate and all 10 essential amino acids were included (Combinations 9 and 18), it was confirmed that the antioxidant effect was significantly improved compared to the cases in which some raw materials were excluded (Combinations 1 to 8, 10 to 17) became Among the peptides, ethyl tetrapeptide-5 appears to play a large role in the antioxidant effect (see Combinations 4 and 13), and the remaining four peptides appear to have a low antioxidant effect. Even in the case of amino acid mix, when it is not included (refer to Combinations 6 and 15), the antioxidant effect is significantly lowered, indicating that it exerts a high antioxidant effect. (see combinations 7, 8, 16 and 17).

<Example 3: Confirmation of collagenase activity inhibitory effect>

In order to confirm the skin wrinkle improvement effect of the composition prepared in Example 1, an in vitro collagenase activity inhibition experiment was performed according to the following method, and the results are shown in Table 4 below.

[Collagenase activity inhibition test method]

(1) Prepare EGCG at the same concentration (0.1% (v/v)) as a positive control.

(2) Prepare a mixture containing a sample (0.1% (v/v)), 100 mM Tris-cl buffer, 100 mM CaCl ₂ , 0.25 mg/ml and collagenase and react.

(2) The reaction is terminated by adding 25 mM citric acid to 400 μl/tube.

(3) Then, 1.5 ㎖/tube of ethyl acetate is added, and after vortexing, the supernatant is taken and transferred to sodium sulfate 150 mg/ep-tube, and centrifuged after vortexing (10,000 rpm, 3 minutes).

(4) Take 300 μl/ep-tube of the supernatant, transfer it to a quartz 96-well plate, measure the absorbance at 320 nm, and calculate the collagenase inhibitory ability according to Equation 3 below.

[Equation 3]

division Collagenase inhibitory ability (%) Control group (untreated group) 0 Combination 1 46.5 Combination 2 37.4 combination 3 35.7 combination 4 44.6 combination 5 42.9 combination 6 45.4 combination 7 49.9 combination 8 50.8 combination 9 70.3 combination 10 42.7 combination 11 22.8 combination 12 22.7 combination 13 39.3 combination 14 34.9 combination 15 32.7 combination 16 32.9 combination 17 39.1 combination 18 60.7 Control (EGCG) 72.4

Referring to Table 4, the collagenase inhibitory ability can be confirmed in all combinations, in particular, according to the present invention, acetyl hexapeptide-8 (Acetyl Hexapeptide-8), hexapeptide-9 (Hexapeptide-9), tripeptide-1 (Tripeptide-1), tetra-acetyl peptide -5 (acetyl Tetrapeptide-5), hexapeptide -11 (hexapeptide-11), Edelweiss callus culture extract (Leontopodium alpinum extract callus culture), bipyridinium the lysate (lysate Bifida Ferment) for fermentation And when all 10 essential amino acids were included (Combinations 9 and 18), it was confirmed that some raw materials were excluded (Combinations 1 to 8, 10 to 17), which showed a significantly improved collagenase inhibitory effect. Among the peptides, hexapeptide-9 and hexapeptide-11 showed high collagenase inhibitory ability (refer to Combinations 2, 3, 11 and 12), and the remaining 3 peptides and amino acid mix, Bifida fermentation lysate and Edelweiss callus culture extract It is also confirmed that there is a collagenase inhibitory ability.

<Example 4: MMP-1 (Matrix Metalloproteinase-1) production inhibition test>

Substances that inhibit the production of Matrix methaloprotease-1 (MMP-1, collagenase) protect collagen and maintain the mechanical properties of the skin tissue to prevent elasticity and skin sagging. Therefore, it can be evaluated that active ingredients can be usefully used in the development of cosmetics for wrinkle improvement and elastic skin. Accordingly, the MMP-1 production inhibitory ability test of the compositions shown in Table 1 was performed.

_{Human dermal fibroblasts (HDF) cells were cultured at 37° C. and 5% CO 2 in} a medium supplemented with 10% fetal bovine serum (Sigma) in DMEM (Dulbecco's modified Eagle's media, Sigma). . After culturing cells in a 24-well plate at ^{a concentration of 2 × 10 5} cells/well and confirming cell adhesion, the untreated control group (CTL) was treated with nothing and only a solvent was added, and 18 samples (combination 1) were added to the experimental group. to 18) and as a positive control, retinol was treated to a concentration of 10 μg/ml, and acetyl hexapeptide-8 (Acetyl Hexapeptide) was treated for efficacy comparison between the composition according to the present invention and each component constituting it. -8), hexapeptide-9 (Hexapeptide-9), tripeptide-1, acetyl tetrapeptide-5 (Acetyl Tetrapeptide-5) and hexapeptide-11 each 0.05% ( w/v), Edelweiss callus culture extract 10% (v/v), Bifida fermentation lysate 10% (v/v) and Essential amino acid Mix 10% (w/v) were treated alone.

After the cells were treated with each of the above substances, they were cultured for 2 days. After 2 days, the medium was collected and the ability to inhibit the production of MMP-1 was measured using Matrix Metalloproteinase-1 (MMP-1), Human, and biotrak ELISA kit (GE healthcare RPN2610), and the results are shown in FIG. 1 .

As shown in FIG. 1 , it was confirmed that the expression level of MMP-1 produced in cells was decreased in most of the experimental groups. In particular, according to the present invention, Acetyl Hexapeptide-8, Hexapeptide-9, Tripeptide-1, Acetyl Tetrapeptide-5, When Hexapeptide-11, Leontopodium alpinum callus culture extract, Bifida Ferment Lysate and all 10 essential amino acids are included (Combinations 9 and 18) Compared to the excluded cases (Combinations 1 to 8, 10 to 17), the effect was significantly superior, confirming that there is a synergistic effect between the combined raw materials. In particular, even when the concentration of the total sum of peptides was treated the same at 0.05% (w/v), the combination of five peptides could confirm the highest activity. Among the peptides, it was found that hexapeptide-9 and acetyl hexapeptide-8 had high MMP-1 expression inhibition ability, and in the case of Edelweiss callus culture extract, Bifida fermentation lysate and 10 essential amino acids, MMP-1 production inhibitory effect was found to be low.

From these results, when UV that induces MMP-1 production on the skin is applied as a factor, according to the present invention, Acetyl Hexapeptide-8, Hexapeptide-9, Tripeptide-1 (Tripeptide-1), tetra-acetyl peptide -5 (acetyl Tetrapeptide-5), hexapeptide -11 (hexapeptide-11), Edelweiss callus culture extract (Leontopodium alpinum extract callus culture), bipyridinium the lysate (lysate Bifida Ferment) for fermentation And it is confirmed that using a mixture of 10 essential amino acids has an excellent effect of suppressing wrinkles on the skin and can help skin aging.

<Example 5: Evaluation of intracellular collagen production ability>

The intracellular collagen production ability test was performed on the 18 compositions that showed an effect in the MMP-1 production inhibitory ability test. Human dermal fibroblasts (HDF) to be used in the test are medium 106 (cascade biologics, M-106-500) added with 1X LSGS (Low Serum Growth Supplement, cascade biologics S-003-10). 37 C and incubated at 5% CO _{2 conditions.} Cells were cultured in a 24-well plate at ^{a concentration of 1 X 10 5} cells/well and cell adhesion was confirmed. In the untreated control group (CTL), nothing was treated and only a solvent was added, and 18 samples (combinations) were added to the test group. 1 to 18) and TGF (Tumor growth factor) as a positive control was treated to a concentration of 10 ㎍ / ㎖. In addition, in order to compare the efficacy of the composition according to the present invention and each component constituting the same, Acetyl Hexapeptide-8, Hexapeptide-9, Tripeptide-1 1), acetyl tetrapeptide-5 (Acetyl Tetrapeptide-5) and hexapeptide-11 (Hexapeptide-11) each 0.05% (w/v), Edelweiss callus culture extract 10% (v/v), Bifida fermentation Lysate 10% (v/v) and Essential Amino Acid Mix 10% (w/v) were treated alone.

After the cells were treated with each of the above substances, they were cultured for 2 days. After 2 days, the medium was collected and the amount of collagen produced was measured using a Procollagen Type I C-peptide (hereinafter PIP) EIA kit (Takara MK101), and the results are shown in FIG. 2 .

Referring to FIG. 2 , as a result of measuring the amount of collagen produced after treatment with 18 experimental groups, it was confirmed that the amount of collagen increased in all experimental groups compared to the control group (CTL). Among these test groups, in particular, according to the present invention, acetyl hexapeptide-8 (Acetyl Hexapeptide-8), hexapeptide-9 (Hexapeptide-9), tripeptide-1 (Tripeptide-1), acetyl tetrapeptide-5 (Acetyl Tetrapeptide-5), hexapeptide -11 (hexapeptide-11), Edelweiss callus culture extract (Leontopodium alpinum extract callus culture), bipyridinium the case with all fermentation lysate (Bifida Ferment lysate), and essential amino acids for the paper 10 (a combination 9 and 18) Compared with cases where some raw materials were excluded (Combinations 1 to 8, 10 to 17), the effect was significantly superior, confirming that there is a synergistic effect between the combined raw materials.

In particular, even when the concentration of the total sum of peptides was treated the same at 0.05% (w/v), the combination of five peptides could confirm the highest activity. In addition, it was confirmed that tripeptide-1, edelweiss callus culture extract and bifida fermentation lysate among peptides had excellent collagen production promoting ability, and 10 essential amino acids had low collagen production promoting effect.

From these results, collagen production in the skin was reduced compared to the case of using a single raw material, Acetyl Hexapeptide-8, Hexapeptide-9, Tripeptide-1, Acetyl Contains tetrapeptide-5 (Acetyl Tetrapeptide-5), hexapeptide-11 (Hexapeptide-11), Leontopodium alpinum callus culture extract, Bifida Ferment Lysate and all 10 essential amino acids It is confirmed that using the prepared composition shows more useful results and can suppress wrinkles on the skin by increasing collagen production.

<Example 6: Skin cell regeneration ability test>

The purpose of this study was to verify the cell regeneration-inducing effect of functional peptides through wound healing assay, an experiment that can promote the growth of damaged skin cells and improve cell recovery.

Human keratinocytes (HACAT, Human Keratinocyte) to be used in the test were cultured at 37° C. and 5% CO ₂ in DMEM medium containing 10% FBS. The middle of the human skin keratinocyte line grown to 100% fill in a 6 well plate was uniformly wounded using a tip, washed twice with PBS to remove floating cells, and the composition of Example 1 Combination 9 was added to the medium. Treated to be 10, 5 and 2.5% (v/v), respectively.

The experimental time is based on a total of 24 and 48 hours, and the degree of skin cell regeneration was confirmed by observing the degree of recovery of the wound during each time through an image device, and the results are shown in FIG. 3 .

Referring to Figure 3, compared to the untreated control (Control) according to the present invention acetyl hexapeptide-8 (Acetyl Hexapeptide-8), hexapeptide-9 (Hexapeptide-9), tripeptide-1 (Tripeptide-1), acetyl Contains tetrapeptide-5 (Acetyl Tetrapeptide-5), hexapeptide-11 (Hexapeptide-11), Leontopodium alpinum callus culture extract, Bifida Ferment Lysate and all 10 essential amino acids In the composition (Combination 9), it was confirmed that cell growth was active in a concentration-dependent manner, and it was observed that the wound healing effect of skin cells was excellent. Through this, it was found that the present anti-aging composition has an effect of inducing cell regeneration.

<Example 7: Stability confirmation experiment of anti-aging cosmetic composition>

In order to confirm the thermal and light stability of the composition of the present invention, the composition (combination 9 and 18) was dissolved in 50 mM Tris-HCl (pH 8.0) buffer to a concentration of 10 mg/ml and dispensed into a glass vial. After storage at room temperature for 7, 14, 21 and 28 days, the loss of peptides and growth factors in the composition due to heat was measured, and stability under sunlight conditions was measured in the same way, and the results are shown in FIGS. 4 and 5 indicated.

4 and 5 , as a result of measuring the loss of the peptide due to heat and measuring the stability in sunlight conditions, it was confirmed that the peptide had a low loss of less than 10% and had high thermal and light stability.

<Example 8: Cytotoxicity test>

In order to determine whether the composition of the present invention exhibits toxicity to keratinocytes, MTT assay was performed. HACAT cells and Human Dermal Fibroblast (HDF) cells were ^{seeded to a cell number of 4 X 10 4} cells/24 well per well in a 24 well plate, and then incubated at 37° C. and 5% CO ₂ conditions for 24 hours. After washing twice with PBS, the composition (Combination 9) was treated with each concentration (20 to 2.5% (v/v)) and incubated for 24 hours. After incubation, MTT 0.5% DPBS was mixed with the culture medium at 1:9 (v/v) and added, followed by _{incubation in a CO 2} incubator for 2 hours. Then, the generated formazan was dissolved in DMSO and measured at 570 nm using ELISA. , the results are shown in FIGS. 6 and 7 .

6 and 7, with respect to HaCat cells, which are a type of keratinocytes, and NIH3T3 cells, which are a type of fibroblasts, a decrease in the number of cells by the composition treated from a low concentration to a high concentration was not observed, and the cells were observed under a microscope. No significant change was observed either.

Through this, it can be seen that the composition according to the present invention has no toxicity to skin-related cells, and thus it can be predicted that the composition will not have a significant effect even if the composition is treated on the skin.

<Preparation Example: Preparation of cream-type composition containing anti-aging composition>

A cream-type composition containing the anti-aging composition was prepared by the following method. As a cream-type composition, 10 parts by weight of the composition of Combination 9 of Example 1 was included to constitute a formulation as shown in Table 5 below.

1) Preparation of [Aqueous B]

Water is heated and stirred at 60 to 70° C., and powdered raw materials are sequentially added, stirred with a disper mixer until it thickens, and then evenly dissolved, then cooled, and prepared in advance for process easiness.

2) Preparation of [Aqueous A]

After the powdery raw material is sufficiently impregnated with glycerin, water is added, and the mixture is heated to 50 to 60° C. and uniformly dissolved with a disper mixer.

3) [Water A] + [Water B] Mix Stirring Preparation

Add pre-prepared aqueous phase B to aqueous phase A and warm while mixing evenly. Prepare to be maintained at 65 to 75 ℃.

4) [Oil phase] Mixed and stirred manufacturing

Each raw material is weighed in an individual beaker and melted by heating and stirring (over 80℃). The molten phase is prepared so that the temperature can be maintained above 75°C.

5) Whole cream preparation

The oil phase is slowly added to the prepared aqueous mixture and stirred vigorously (6,000 rpm) for about 10 minutes to proceed with the emulsification process. At this time, the temperature is to be maintained at 65 to 75 ℃. After the emulsification process is completed, the additive A is prepared and the pH adjustment and neutralization process is performed. At this time, since the viscosity of the contents rises, it is stirred vigorously (6,000 rpm) evenly for 5 minutes or more. When the contents cool down to 50 ° C or less, add Additive B and mix slowly for about 2 minutes (3,000 to 4,000 rpm). When the temperature is below 40 ° C, add Additive C sequentially and stir slowly for about 2 minutes (3,000 to 4,000 rpm), The contents are collected through a degassing/filtration process. After completion of the degassing/filtration process, pH adjustment and neutralization process are performed. At this time, since the viscosity of the contents rises, stir vigorously (6,000 rpm) evenly for 5 minutes or more. When the temperature is below 40 ℃, the anti-aging composition (PEPTamino-C) and fragrance (additive D) are added, stirred slowly for about 2 minutes (3,000-4,000 rpm), and then the contents are collected through a degassing/filtration process.

division Ingredients / Ingredients Ingredients / INCI name content
(parts by weight) Award A DI WATER Water 38.14 GLYCERIN Glycerin 10.00 Keltrol F Xanthan Gum 0.10 Adenosine Adenosine 0.04 Award B DI Water Water 30.00 Hyaluronate Acid 0.2% Sodium Hyaluronate 0.20 Carbopol 940 Carbomer 0.02 paid Kalcol 6870 Cetearyl Alcohol 2.50 GMS-105 Glyceryl Stearate 1.50 Arlacel 165V Glyceryl Stearate, Peg-100 Stearate 1.20 Tween 80 Polysorbate 80 1.00 Arlacel 83 Sorbitan Sesquioleate 0.70 Phytosqualane Squalane 5.00 DUB MCT Caprylic/Capric Triglycerides 3.00 Additive A DI Water Water 3.00 L-Arginine (pH 4.50 ~ 5.50) Arginine 0.50 Additive B Symdiol 68 Caprylyl Glycol & 1,2 Hexanediol 2.00 Additive C Lutein Solution Lutein 1.00 Additive D (fragrance) Perfume Fragrance 0.10 anti-aging composition PEPTamino-C Combination 9 composition 10.00

Preferred embodiments of the present invention described above are disclosed to solve the technical problem, and various modifications, changes, additions, etc. will be possible within the spirit and scope of the present invention by those skilled in the art to which the present invention pertains. , such modifications and changes should be regarded as belonging to the following claims.

The composition according to the present invention is very useful for manufacturing functional foods, cosmetics, and pharmaceuticals by preventing aging of the skin and keeping the skin healthy and young through the anti-oxidation effect, the wrinkle improvement effect and the cell regeneration effect due to the promotion of collagen production and decomposition inhibition. It can be widely used, so it has very high industrial applicability.

Claims (10)

Acetyl Hexapeptide-8, Hexapeptide-9, Tripeptide-1, Acetyl Tetrapeptide-5, Hexapeptide-11 -11), Edelweiss callus culture extract ( Leontopodium alpinum callus culture extract), Bifida Ferment Lysate, Isoleucine, Valine, Leucine, Arginine, Lysine ), phenylalanine (Phenylalanine), methionine (Methionine), histidine (Histidine), tryptophan (Tryptophan) and threonine (Threonine) anti-aging composition comprising. According to claim 1,
Compared to the total composition, the acetyl hexapeptide-8 (Acetyl Hexapeptide-8), hexapeptide-9 (Hexapeptide-9), tripeptide-1 (Tripeptide-1), acetyl tetrapeptide-5 (Acetyl Tetrapeptide-5) and hexapeptide -11 (Hexapeptide-11) is 0.001 to 1.0% (w/v), respectively, and the edelweiss callus culture extract (Leontopodium alpinum callus culture extract) and Bifida Ferment Lysate are each 0.5 to 20% (v) /v), the isoleucine, valine, leucine, arginine, lysine, phenylalanine, methionine, histidine, tryptophan, and Threonine is an anti-aging composition, characterized in that it is contained in an amount of 0.1 to 10% (w/v), respectively. According to claim 1,
The edelweiss callus culture extract is an anti-aging composition, characterized in that prepared according to the following method:
(a) separating the cambium from the stem internal tissue after disinfecting and peeling the epidermis of the edelweiss stem, inoculating the edelweiss tissue into a medium to induce callus and then proliferating;
(b) inoculating the induced callus in a bioreactor and culturing; and
(c) extracting by adding a solvent after drying the cultured callus. According to claim 1,
The bifida fermentation lysate is B. actinocoloniiforme, B. adolescentis, B. angulatum, B. animalis, B. aquikefiri, B. asteroides, B. biavatii, B. bifidum, B. bohemicum, B. bombi, B. boum , B. breve, B. callitrichos, B. catenulatum, B. choerinum, B. commune, B. coryneforme, B. cuniculi, B. crudilactis, B. denticolens, B. dentium, B. eulemuris, B. faecale, B Gallicum, B. gallinarum, B. hapali, B. indicum, B. inopinatum, B. kashiwanohense, B. lemurum, B. longum, B. magnum, B. merycicum, B. minimum, B. mongoliense, B. moukalabense , B. myosotis, B. pseudocatenulatum, B. pseudolongum, B. psychraerophilum, B. pullorum, B. reuteri, B. ruminantium, B. saguini, B. scardovii, B. stellenboschense, B. stercoris, B. saeculare, B. subtile, B. thermacidophilum, B. thermophilum,,B. tissieri and B. tsurumiense An anti-aging composition, characterized in that it is cultured and fermented by inoculating a microorganism of the genus Bifidobacterium selected from the group consisting of. 5. The method according to any one of claims 1 to 4,
The composition is an anti-aging composition, characterized in that it has the ability to promote skin cell regeneration through the promotion of human skin cell growth. 5. The method according to any one of claims 1 to 4,
The composition is an anti-aging composition, characterized in that it has the ability to improve skin wrinkles by promoting collagen production in human fibroblast lines and inhibiting collagen degrading enzyme (MMP-1) activity. 5. The method according to any one of claims 1 to 4,
The composition is an anti-aging composition, characterized in that it has antioxidant activity through the removal of active oxygen. A cosmetic composition comprising the composition of any one of claims 1 to 4. A pharmaceutical composition comprising the composition of any one of claims 1 to 4. A functional food composition comprising the composition of any one of claims 1 to 4.

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