KR20210066537A - A composition for improving skin beauty containing glycosaminoglycan extracted from insect - Google Patents

Abstract

The present invention relates to a composition for promoting skin beauty containing insect-derived glycosaminoglycan as an active ingredient and, more particularly, to a composition for promoting skin beauty, such as improving skin elasticity and moisture, alleviating skin wrinkles, improving a skin barrier function, and helping anti-inflammation by containing mucopolysaccharides extracted from bees, in particular, glycosaminoglycans extracted from male bees of Bombus terrestris as well as a bee colony. According to the present invention, insect-derived glycosaminoglycans are all excellent in inhibiting elastase activity, inhibiting collagenase activity, and promoting hyaluronic acid production, are confirmed to be safe for human skin, and have excellent effects, especially for skin moisturizing, skin wrinkle reduction, or the like.

Description

A composition for improving skin beauty containing insect-derived glycosaminoglycan as an active ingredient {A composition for improving skin beauty containing glycosaminoglycan extracted from insect}

The present invention relates to a composition for enhancing skin beauty containing insect-derived glycosaminoglycan as an active ingredient, and more particularly, by containing glycosaminoglycan derived from bee and bee groups, which are mucopolysaccharides extracted from bees. , elastin decomposition and collagen decomposition, and greatly increases the concentration of hyaluronic acid to help improve skin elasticity and moisture and improve skin wrinkles, strengthen the skin barrier function, and inhibit the activity of inflammatory mediators to have anti-inflammatory effects It relates to a composition for enhancing skin beauty.

The skin is an epithelial tissue that covers and protects the muscles and organs of the body. It is composed of the epidermis, dermis, and subcutaneous fat. It protects the body from the external environment, regulates body temperature, and allows you to feel sensations, mainly from external physical damage and chemicals. It protects the human body, prevents bacteria, fungi, and viruses from penetrating the skin, and acts as a protective barrier to prevent moisture loss. Among them, the epidermis is the thinnest of the three layers, and it plays an important role in moisturizing and protecting the skin. In particular, the moisture in the stratum corneum of the epidermis makes the skin elastic and soft.

In addition, in recent years, cosmetics having whitening, skin wrinkle improvement and suppression, and UV protection functions are continuously being developed to satisfy consumers' desire for high-performance cosmetics. Among them, wrinkle improvement and suppression products in particular keep the skin young. It is an object that attracts a lot of attention from consumers who want to

As we age, collagen deficiency or quality deteriorates, causing the skin to lose elasticity and wrinkles to form. Skin wrinkles are affected by several factors such as the moisture content, collagen content, and immune function of the skin, and the elasticity of the skin is this It is known to be maintained by collagen and elastin (elastic fibers).

In vivo, the synthesis and degradation of extracellular matrix such as collagen is properly regulated, but as aging progresses, its synthesis decreases and the expression of matrix metalloproteinase (MMP), an enzyme that decomposes collagen, is promoted, so that the skin The elasticity of the skin is reduced and wrinkles are formed.

In addition, macrophages are known to be involved in the maintenance of homeostasis by participating in various host reactions, and in the case of an inflammatory reaction, they are caused by nitric oxide (NO, nitric oxide) and cyclooxygenase-2 (COX-2, cyclooxygenase-2). It produces pro-inflammatory factors such as prostaglandin E2 (PGE2, prostaglandin E2), which are produced in excess. Therefore, substances that directly inhibit the activity of PGE2 inflammatory mediators are highly likely to be developed as anti-inflammatory agents that inhibit various inflammation-related diseases such as acne, atopic dermatitis, rheumatoid arthritis, and malignant neoplasms. Many studies are underway to find an anti-inflammatory treatment from various natural foods and herbal medicines that are safer and more effective to substitute for.

On the other hand, glycosaminoglycan (GAG) has a structure in which uronic acid and glucosamine are repeated and partially sulfated, and can exhibit various kinds of biological activities by this complex chemical structure. According to what is known so far, these various biological activities appear to be due to the presence of unique specific sequences. GAGs exist in the form of proteoglycans in vivo, and these proteoglycans are decomposed by proteases and glycosidases to release sugars. Although the physiological roles of GAGs have not been elucidated yet, they appear to exhibit physiological activity by binding to proteins in the living body.

Such GAG can be extracted from various insects, but it is known that the extraction amount and GAG composition are different depending on the type of insect, and a method for preparing GAG from insects is also proposed in Korean Patent Registration No. 10-1522604.

According to the prior art, it can also be extracted from the hornbeam bee, Bombus terrestris ) is an insect of the Hymenoptera family, which is native to Europe and was introduced as a resource insect (for pollinator) in Korea. The length of the queen bee is 20~22mm, that of the male bee is 14~16mm, and that of the worker bee is 11~17mm.

In addition, the method for producing GAG from insects and the use of anti-inflammatory, antidiabetic, anticoagulant, anticancer, angiogenesis inhibitory, and/or pharmaceutical use for the treatment and/or prevention of diseases caused by dysfunction of vasodilation reactions are disclosed in Korean Patent Laid-Open No. 10 - It is disclosed in 2011-0013074 (a novel method for preparing insect glycosaminoglycans), and thereafter, a liver protection food composition (Korean Patent Registration No. 10-1869792) and anti-inflammatory and arthritis improvement Application to health functional food (Korean Patent Registration No. 10-1680644), etc. has been disclosed.

However, this existing technology for using GAG has not yet been commercialized as a product with a clear therapeutic effect, so research for practical use continues. I have a problem that I can't.

In particular, in the prior art, there is a lack of technology to utilize the function as an active ingredient by itself in utilizing GAG, so it is only to the extent that there is an attempt to use it for various purposes by mixing it with other carriers or active ingredients.

Therefore, in the prior art, there was a problem in that the usefulness of GAG as an active ingredient was not properly revealed.

Korean Patent No. 10-1522604 Laid-Open Patent Publication No. 10-2011-0013074 Registered Patent Publication No. 10-1869792 Registered Patent Publication No. 10-1680644

In order to solve the problems of the prior art and the problem of improving usability, the present invention makes the development of an effective new use using GAG itself an active ingredient as a solution.

To solve this problem, the present inventors have been continuously conducting research to develop new uses of GAG, GAG, especially insect-derived GAG, inhibits elastin degradation and collagen degradation, and increases hyaluronic acid concentration to improve skin elasticity and moisture And by helping to improve wrinkles and strengthening the skin barrier function, as well as having an anti-inflammatory effect by inhibiting the activity of inflammatory mediators, it was found that there is an effect of improving skin beauty and completed the present invention.

Accordingly, an object of the present invention is to provide a composition for promoting skin beauty as a new use of insect-derived GAG.

In addition, another object of the present invention is to provide a cosmetic composition for promoting skin beauty containing insect-derived GAG.

In addition, another object of the present invention is to provide a health food for promoting skin beauty containing insect-derived GAG.

In order to solve the problem of the present invention as described above, the present invention provides a composition for enhancing skin beauty containing insect-derived glycosaminoglycan (GAG) as an active ingredient.

In addition, the present invention provides a cosmetic composition for improving skin wrinkles containing insect-derived glycosaminoglycan (GAG) as an active ingredient.

In addition, the present invention provides a moisturizing cosmetic composition containing insect-derived glycosaminoglycan (GAG) as an active ingredient.

In addition, the present invention provides an anti-inflammatory cosmetic composition containing insect-derived glycosaminoglycan (GAG) as an active ingredient.

In addition, the present invention provides a health food for promoting skin beauty containing insect-derived glycosaminoglycan (GAG) as an active ingredient.

According to a preferred embodiment of the present invention, the insect-derived GAG is preferably contained in an amount of 0.01-5% by weight as an active ingredient in the entire composition for skin beauty promotion.

The composition for improving skin beauty containing insect-derived GAG according to the present invention is safe for the skin and has an anti-inflammatory effect by inhibiting the decomposition of elastin and collagen, thereby improving the elasticity and moisturizing of the skin, and inhibiting the activity of inflammatory mediators. Therefore, it exhibits excellent effects in improving skin beauty such as maintaining skin moisture, improving wrinkles and anti-inflammatory.

In addition, since the GAG according to the present invention exhibits effective activity for skin beauty improvement by itself, unlike the existing ones, it is a composition for skin beauty improvement by itself even if other active ingredients are not mixed and used as a cosmetic or health food. There is a usable effect.

1 is a graph showing the immune cell cytotoxicity of GAGs derived from the hornbeam and bong group used in the Examples of the present invention.
Figure 2A is a graph showing the HAS-2 mRNA expression effect of GAG derived from the hornet bee and bong group used in the Examples of the present invention.
Figure 2B is a graph showing the HAS-3 mRNA expression effect of GAG derived from hornet bee and bong group used in Examples of the present invention.
Figure 3 is a graph showing the filaggrin mRNA expression effect of GAG derived from the hornet bee and bonggun used in Examples of the present invention.
Figure 4 is a graph showing the Elastase inhibitory ability of the GAG derived from the scorpion and bong group used in the Examples of the present invention.
Figure 5 is a graph showing the LPS-induced inflammation inhibitory effect of GAG derived from hornet bee and bonggun used in Examples of the present invention.

Hereinafter, the present invention will be described in more detail as one embodiment as follows.

The present invention relates to glycosaminoglycans known to have anticancer effects; GAG) to new uses.

In particular, the present invention does not simply mix and use insect-derived GAG with other active ingredients, but essentially allows GAG alone to function as an active ingredient, and proposes a new use as an active ingredient for GAG.

In general, GAG is known as a pharmaceutical for anti-inflammatory, anti-diabetic, anti-coagulant, anti-cancer, angiogenesis-inhibiting, and/or disease treatment and/or prevention due to dysfunction of vasodilation, or as a food for inhibiting fat accumulation. However, practical use as an active ingredient for skin beauty promotion has not been proposed at all.

Although many medicines and health foods are being developed, it is one of the difficult tasks to invent that these medicines or food ingredients are effective in promoting skin beauty or improving skin beauty.

In the present invention, as a new effect on skin beauty promotion of the GAG component, which has not previously been found to have an effect for promoting skin beauty, it has been newly found that it is particularly effective in moisturizing and wrinkle improvement of the skin, strengthening the skin barrier function and anti-inflammatory.

The skin is a very sensitive part, and in particular, in the case of facial skin, it is very sensitive compared to other parts, so it is difficult to find a substance that expresses a cosmetic improvement effect on the skin. Since wrinkles of the skin are affected by various factors such as the moisture content, collagen content, and immune function of the skin, it suppresses the breakdown of elastin and collagen and significantly increases the concentration of hyaluronic acid to help improve skin elasticity and moisture By giving it, skin wrinkles can be improved. Inflammation of the skin is caused by excessive production of inflammatory factors such as prostaglandin E2 (PGE2, prostaglandin E2), and inflammation of the skin can be improved by inhibiting the activity of PGE2 inflammatory mediators.

According to the present invention, it was found for the first time that the insect-derived GAG prepared in the present invention inhibits elastin degradation and collagen degradation, significantly increases hyaluronic acid concentration, strengthens skin barrier function, and reduces the activity of PGE2 inflammatory mediators.

Accordingly, the present invention provides a composition for promoting skin beauty containing insect-derived GAG as an active ingredient for inhibiting elastin degradation, inhibiting collagen degradation, increasing hyaluronic acid concentration, strengthening skin barrier function, and reducing PGE2 inflammatory mediator activity. In particular, it shows excellent efficacy when applied as a cosmetic composition for skin wrinkle improvement by using insect-derived GAG as an active ingredient for inhibiting elastin decomposition and collagen decomposition and increasing hyaluronic acid concentration. In addition, it has an excellent effect for moisturizing the skin, and can be applied as a cosmetic for inner beauty or as a health food for skin beauty promotion.

GAG can be obtained by extraction from various natural resources, for example, preferably obtained by extraction from insects. In the present invention, particularly those derived from bees can be preferably used.

According to a preferred embodiment of the present invention, preferably, the insect-derived GAG extract most suitable for the purpose of the present invention is preferably a bee, and among them, GAG obtained from a group of bees and colonies is particularly used for elastin degradation, collagen degradation and It was confirmed that the anti-inflammatory effect was excellent.

Since western bees are widely used as pollinators in rural areas, there are also developed beekeeping companies that supply them, so it has the advantage of being easy to collect. In particular, male bees and colonies are easier to collect than queen bees and they are easy to use because of their large supply. There are advantages.

In addition, the generally discarded bees and colonies can be used, so the purchase cost is low, and it is environmentally preferable because the discarded insect by-products are recycled.

Therefore, as a preferred example of the present invention, GAG extracts from bees, in particular, hornet bees and swarms can be used as an active ingredient in the composition for skin beauty promotion.

In addition, the present invention is preferably for GAG extraction, preferably, for example, washing the honey bee bees and colonies with an organic solvent to remove the honey component and fats, drying the residue, grinding the dried residue to 150 ~ 500㎛ , The pulverized product of the above step is protein-digested with a proteolytic agent and removed, and the protein-removed residue is precipitated in ethanol and washed. Therefore, the present invention includes a method for preparing such a GAG extract.

Glycosaminoglycan (GAG), which can be prepared from the queen bee, contained in the composition for enhancing skin beauty according to the present invention, is disclosed in, for example, Korean Patent No. 10-1522604 (a heparin fragment derived from an insect glycosaminoglycan and Its manufacturing method), but it is preferable to prepare and use it specifically by the following method.

(1) washing bees and colonies with ethanol with ethanol and drying them, followed by primary grinding;

(2) extracting the pulverized product with ethanol to remove fats, recovering the residue, drying it with dry heat, and pulverizing to an average diameter of 150 to 500 μm;

(3) adding 5-15 times the volume of sodium carbonate, preferably pH 8-10, more preferably pH 8.5-9.5 sodium carbonate to the residue above, stirring, and decomposing the protein with a proteolytic enzyme;

(4) Trichloroacetic acid in gastric juice removing the degraded protein after adding acid and inducing precipitation;

(5) precipitating by adding ethanol and sodium acetate to the supernatant, and recovering the precipitate after refrigeration;

(6) Washing the above sediment with ethanol and drying it to obtain hornet bee and colony GAG.

The first step above is not only to simply remove foreign substances, but also to facilitate the removal of fats by primary grinding. In general, many pollen, pollen, and honey components are present in bees and colonies, so it is not easy to recover the residue, so it is preferable to wash in a way to reduce the viscosity by washing with an organic solvent in a gradient. In addition, since insects have a high fat content and thus it is difficult to extract necessary components, removing fats is a very important problem in industry. In the past, there was no special solution for this, and the next step, crushing, was performed. However, the present invention is preferable and in order to solve the problems of the prior art, it removes foreign substances mixed in a lot in the insect collection step, and also reduces the size through primary intermittent grinding to widen the area in contact with the solvent and absorb it into the inside This facilitates the removal of fats very efficiently.

If the average diameter of the residue in step 2 is smaller than 150㎛ (usually 100mesh), it is very difficult to pulverize and the loss rate is high. If it is larger than 500㎛ (usually 35mesh), the enzymatic decomposition may be delayed, more preferably 350~ 500 µm (usually 35 to 45 mesh) is more preferable.

According to the present invention, when pulverizing to a smaller size compared to the present invention in this pulverization process, for example, pulverizing a particle size smaller than 100mesh size, there is a problem in that drying is difficult due to the nature of insects during the manufacturing process and the pulverization process is very difficult. In addition, the process time required for pulverization takes a long time, and there is a concern that the enzymatic decomposition may be delayed because the particles are too small. In addition, since there is a high risk of being introduced into the supernatant during centrifugation, separation is also difficult, which is rather disadvantageous and uneconomical in terms of the process, and acts as a factor for lowering the yield. Therefore, in the present invention, in order to solve this problem, it is preferable to make the average particle diameter of the residue, preferably, the particle diameter larger than 100 mesh.

In the above 3 steps of the present invention, the protease uses, for example, Alcalase (Alkaline Protease), and the reaction can be carried out at a temperature above room temperature for 1 to 7 days, but 2 to 3 days are preferable because it is efficient.

The composition for enhancing skin beauty of the present invention may be used in an amount of 0.01 to 5.0% by weight relative to the total weight of the total composition in order to contain GAG as an active ingredient. If, when the composition for enhancing skin beauty of the present invention is composed of cosmetics, it may be included in an economical aspect, preferably 0.01-1.0% by weight, more preferably 0.03-0.3% by weight. In addition, in order to contain as an active ingredient by applying as an orally administered inner beauty cosmetic or health food among cosmetics, it may be contained in an excess of, for example, 0.3-5.0 wt%. As a preferred example, GAG is preferably contained in an amount of 5-1000 μg/ml, preferably 100-700 μg/ml, more preferably 200-700 μg/ml.

According to the present invention, when GAG is included in less than 0.01% by weight relative to the total weight in order to be used as an active ingredient in the composition for skin beauty, there is a problem in that elastin degradation and collagen degradation inhibition do not appear, and more than 5.0% by weight in the total dose. In some cases, side effects such as skin itching may occur in people with dry skin.

The cosmetic composition of the present invention may be prepared in all formulations conventionally prepared in the art, for example, solutions, suspensions, pastes, gels, creams, lotions, packs, foams, etc., but is not limited thereto. Cosmetics consisting of each of these formulations may further include thickeners, antioxidants, preservatives, skin moisturizers, skin softeners, surfactants, L-menthol, hair conditioning agents, anti-foam agents, fragrances, etc. used in conventional cosmetic compositions. have.

In addition, as a health food for promoting skin beauty, it can be used as a liquid, extract, solid, tablet, powder, granule, capsule, and the like.

When the composition for enhancing skin beauty according to the present invention is used as a cosmetic and the cosmetic formulation is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, cellulose derivative, polyethylene glycol, silicone, bentonite as a carrier component , silica, talc or zinc oxide may be used.

When the formulation of a cosmetic using the composition for skin beauty according to the present invention is a solution or emulsion, a solvent, solvating agent or emulsifying agent may be used as a carrier component, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, Benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan.

When the cosmetic formulation using the composition for enhancing skin beauty according to the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbate A suspending agent such as bitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tracanth may be used.

In addition, the health food for promoting skin beauty according to the present invention may be prepared in various formulations, and additionally other ingredients may be combined and used.

Hereinafter, the present invention will be described in more detail through examples, but this is for explaining the present invention in more detail, and the scope of the present invention is not limited thereto.

Preparation Example: Preparation of Glycosaminoglycan (GAG)

Using a sieve, 1 kg of bee and colony groups were used, washed with 100%, 90%, and 70% ethanol in a gradient of ethanol, dried in a Dry-Oven, and then pulverized intermittently, and the above pulverized product was washed with 3 times the volume of ethanol. After extraction, the extraction residue was dried in a Dry-Oven and pulverized to 35 mesh (500 μm) with a pulverizer.

After adding and stirring 10 times the volume of sodium carbonate (pH 9.0) to 100 g of the pulverized residue, 1/10 of the weight of the residue was added with Alcalase powder and cultured for 2 days.

The culture was filtered through a Buchner panel, and trichloroacetic acid was treated to a final concentration of 5% for inactivation of proteins and enzymes present in the solution, and filtered through a Buchner panel.

Two-fold volume of ethanol was added to the solution, and sodium acetate was added to make a final concentration of 1% to precipitate the solution, and after refrigerated storage for 1 day, only the precipitate was collected by filtration through a Buchner panel.

The recovered sediment was washed with 100 → 70% ethanol and then dried with hot air to obtain locust beetle and colony GAG, and the final yield was 1.6% compared to the original material.

Comparative Preparation Example: Preparation of Glycosaminoglycan (GAG)

Among the methods of the preparation example, washing and intermittent grinding of the locust wasp groups and intermittent grinding were not carried out, and after drying, it was prepared in the same manner except that it was pulverized to 200 mesh with a grinder, and the final yield was 0.3% compared to the raw material.

Experimental Example: Evaluation of the skin beauty enhancement effect of glycosaminoglycan (GAG)

(1) Cytotoxicity evaluation of hornbeam bee and colony GAG

Phosphorus RAW264.7 cells derived from macrophages were seeded in a 96-well plate at a concentration of 2×10⁴ cells/well, and cultured for 12 h to allow the cells to completely adhere. , 50, 100, 200, 400 μg/mL) for 24 h. After incubation, formazan formed in living cells by treatment with MTT reagent was dissolved by adding 100 μL of dimethyl sulfoxide (DMSO) per well, and then absorbance was measured at a wavelength of 540 nm.

The results of confirming the cell viability using the MTT assay are shown in FIG. 1 , and cytotoxicity was not observed compared to the control at all concentrations.

(2) Moisturizing effect evaluation of hornwort bee and colony GAG

Ct계산하여 그 결과를 도 2A 및 도 2B에 나타내었다. HaCaT cells, which are human keratinocytes, ^{were dispensed at a concentration of 1×10 6} cells/well in a 6-well plate and cultured for 24 h, then replaced with a serum-free medium and starvated for 24 h. After treatment with bee and colony GAG (0.1, 1, 10 ug/mL), total RNA is extracted and quantified using TRIzol cells, and cDNA is synthesized from 1 ug of RNA using oligo(dT) primer. The synthesized cDNA was subjected to real-time PCR using HAS-2, HAS-3, Filaggrin and GAPDH primers. The experimental results obtained through real-time PCR are based on the housekeeping gene GAPDH.

Ct was calculated and the results are shown in FIGS. 2A and 2B.

As a result, compared to the control group, the expression levels of HAS-2 increased by 1.3 and 1.6 times at 1 ug/mL and 10 ug/mL, respectively, and HAS-3 increased by 1.4 and 2.7 times at 1 ug/mL and 10 ug/mL, respectively. could see that

(3) Evaluation of the skin barrier function of hornet bee bee and colony GAG

In order to investigate the effect of locust wasp and colony GAG on the improvement of skin barrier function, HaCaT cells were treated with bristle wasp and colony GAG as in (2) and cultured for 24 h, followed by RT-PCR expression of filaggrin mRNA The change was confirmed and shown in FIG. 3, and as a result, it increased 2.1 times at 1 ug/mL and increased by 3.0 times at 10 ug/mL.

Therefore, it was confirmed that the function of the skin barrier was strengthened by increasing the expression of filaggrin, a peptide constituting the skin barrier, when treated with the keratinocytes of hornbee bee and mongoose GAG.

(4) Elastase activity inhibitory ability evaluation

In order to evaluate the skin beauty enhancement (wrinkle improvement) effect of GAG, adenosine was used as a comparative substance to measure Elastase activity inhibition. did.

220 μl of sample diluted by concentration (125 μg/ml, 250 μg/ml, 500 μg/ml) in 0.1M Tris-HCl buffer (pH 8.0) and 2 mg/ml N-succinyl-Ala-Ala-Ala- 20 μl of ρ-nitroanilide was added and reacted at 25° C. for 10 minutes. After adding 20 µl of 1 µg/ml porcine pancreatic elastase (PPE) and reacting at 25°C for 20 minutes, the reaction was terminated on ice and absorbance was measured at 405 nm using a microplate reader. and the absorbance reduction rate of the untreated group.

[Equation]

Inhibitory ability (%) = 1 - (absorbance of sample added / absorbance of no-addition) ×100

As a result of the evaluation of the Elastase inhibitory ability, as shown in FIG. 4 , GAG exhibited an Elastase inhibitory activity close to about 2 times at 500 μg/ml compared to adenosine, a comparative material.

(5) Evaluation of the anti-inflammatory efficacy of hornbeam bee and colony GAG

For the evaluation of anti-inflammatory efficacy, the inhibitory ability of prostaglandin E2 ((PGE2, Prostaglandin E2) was measured as follows.

After dispensing RAW 264.7 cells in a 48-well plate at a concentration of 2*10⁴ cells/well, incubate for 12 h to allow the cells to completely adhere, and GAGs in the colony beetle and colony were added at a concentration of 25, 50, 100, and 200 μg/mL. was treated for 12 hours and 1 μg/ml of lipopolysaccharide (LPS, lipopolysaccharide) was treated and cultured for 24 hours. Then, the culture supernatant was taken and the content of PGE2 in the supernatant was measured. The content of PGE2 was measured by the method presented in the manual of ELISA kits (abcam Inc.), and the result was shown in FIG. 5 .

As a result of the measurement, compared with the LPS alone group (1664.8±127.5 pg/mL), it was found to be reduced by 13% to 1447.9±69.6 pg/mL in the group treated with 100 μg/mL GAG of the locust beetle and the group GAG, 200 μg/mL. It was found to be reduced by 26% to 1227.9±83.2 pg/mL in the mL treatment group, and was reduced by 51% to 808.69±128.1 pg/mL in the 400 μg/mL treatment group. Therefore, it is judged that the activity of inhibiting the inflammatory response induced by LPS is very high in the hornbee bee and the mongoose GAG.

As described above, it was confirmed that the hornbee bee and brood GAG have almost no cytotoxicity, and have an excellent effect on skin beauty by having moisturizing, wrinkle, skin barrier function improvement and anti-inflammatory effects.

Example 1: Preparation of cosmetic (gel) for skin beauty enhancement containing GAG

A gel having the following composition was prepared in the same manner as in the general gel preparation method. As GAG, those prepared in the above production example were used.

Raw material content (wt%) Alcohol
glycerin
liquid paraffin
diethoxyethyl succinate
chlorphenesin
triethanolamine
Phenyl Trimethicone
copolymer
Disodium EDTA
Butylene Glycol
Purified water
Licorice root extract
GAG 1.0
2.0
8.0
0.5
0.3
0.5
0.5
0.2
0.1
3.0
83.5
0.1
0.3 Sum 100

Example 2: Preparation of cosmetic (cream) for skin beauty enhancement containing GAG

A cream having the following composition was prepared in the same way as a general cream preparation method.

Raw material content (wt%) Rose Premium Water
Jojoba Golden
argan oil
rosehip oil
macadamere nut oil
olive oil wax
Monta Wax 68
Sea Buckthorn Oil
ceramide
Eoseongcho Extract
Licorice root extract
Napri
GAG 56.5
10.0
6.0
6.0
6.0
5.0
2.0
2.0
2.0
2.0
1.0
1.0
0.5 Sum 100

Experimental Example 2: Safety evaluation of GAG

For the safety test, a sample having a concentration of 600 μg/ml of GAG prepared in Preparation Example was used.

(1) Sensitive skin safety test

Subjects judged to be sensitive in the lactic acid stinging test were selected and conducted according to the standards applied by the International Contact Dermatitis Association and the American Cosmetics Association. It was observed after 48 hours. As a result of the test, the honey bee bee and the colony GAG were judged to be non-irritating substances in terms of the primary irritation test for sensitive skin.

(2) Human skin phototoxicity test

It was tested according to the 24hours Single Patch Test according to the guidelines of the American Cosmetic Association, and at 20 minutes, 24 hours, and 48 hours after light (UVA) irradiation, the skin according to the Personal Care Products Council guidelines (2014) Responses were evaluated by comparison. As a result of the test, 20% of cases had fine erythema, which disappeared within 48 or 72 hours. Therefore, it was judged as safe substances in terms of phototoxicity of human skin.

(3) Eye irritation evaluation test

Tests and judgments were carried out according to the European Hazardous Substances Control Guidelines of the European Animal Substitute Test Method Verification Center, and the average reactivity according to the criteria was 0.1 (0.3-0.4) as a result of the test. Therefore, the level of irritation of the eye mucosa was judged to be in the non-irritating category for the honey bee bee and the colony GAG.

As described above, according to the present invention, it was confirmed that the hornbee bee and bongun GAG can be used as an active ingredient in cosmetics because the safety of the human skin and eye mucosa has been confirmed.

The composition for promoting skin beauty containing insect-derived GAG as an active ingredient according to the present invention has almost no cytotoxicity, and has been confirmed to have moisturizing, wrinkle, skin barrier function improvement and anti-inflammatory effects, and is confirmed to be a safe material for human skin to promote skin beauty It can be used as a cosmetic for skin care or as a health food for skin beauty promotion.

Claims (8)

A composition for enhancing skin beauty containing insect-derived glycosaminoglycan (GAG) as an active ingredient.
A composition for enhancing skin beauty containing 0.01-5% by weight of glycosaminoglycan (GAG) extracted from bees as an active ingredient.
The method according to claim 1, wherein the insect-derived glycosaminoglycan (Glycosaminoglycan, GAG) is a composition for promoting skin beauty, characterized in that extracted from the bee and bee of the hornbeam.
The method according to claim 1, Glycosaminoglycan (Glycosaminoglycan, GAG) is
The step of first grinding after washing and drying the bees and colonies with ethanol;
extracting the above primary pulverized product with ethanol to remove fats, drying the residue and pulverizing it to a diameter of 150 to 500 μm;
10 times the volume of sodium carbonate was added to the residue and stirred, followed by decomposing the protein with a protease;
Trichloroacetic acid in gastric juice removing the degraded protein after adding acid and inducing precipitation;
Recovering the precipitate after precipitation by adding ethanol and sodium acetate to the supernatant and refrigerated storage;
Washing the gastric precipitate with ethanol and drying to obtain glycosaminoglycans derived from hornet bee and colony;
A composition for promoting skin beauty, characterized in that it is prepared with
[Claim 2] The composition for enhancing skin beauty according to claim 1, wherein the washing of the beetle and the colony with ethanol is washing the colony and the colony with ethanol in a gradient of 100%, 90%, and 70% ethanol.
A cosmetic composition for improving skin wrinkles containing insect-derived glycosaminoglycan (GAG) as an active ingredient.
A cosmetic composition for moisturizing containing insect-derived glycosaminoglycan (GAG) as an active ingredient.
A health food for promoting skin beauty containing insect-derived glycosaminoglycan (GAG) as an active ingredient.

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