KR102248170B1 - Composition for skin regeneration and wound healing comprising the extract of actinidia chinensis stem - Google Patents

Abstract

The present invention relates to a composition for skin regeneration or wound healing containing a stem extract of Actinidia arguta as an active ingredient. More particularly, it is characterized in that a composition contains an extract of Actinidia arguta as an active ingredient. According to the present invention, as a cosmetic composition or quasi-drug composition derived from a natural product, there are few side effects, and the skin keratinocyte proliferation and migration promoting effect, collagen production promoting effect, and anti-inflammatory effect are excellent.

Description

A composition for skin regeneration or wound healing that contains the extract of red snail stem extract as an active ingredient {COMPOSITION FOR SKIN REGENERATION AND WOUND HEALING COMPRISING THE EXTRACT OF ACTINIDIA CHINENSIS STEM}

The present invention relates to a composition for skin regeneration or wound healing that contains a blue snail stem extract as an active ingredient, and more particularly, it has excellent proliferation and migration effects of keratinocytes, and has excellent collagen production promotion and anti-inflammatory effects. It relates to a cosmetic composition or a pharmaceutical composition for external use of the skin that can be used for skin regeneration or wound healing.

The skin is in direct contact with the external environment and acts as a barrier to protect the body from various external harmful factors. The skin is the widest organ covering the entire body surface, and is composed of epidermis, dermis and subcutaneous tissue, and the subcutaneous tissue is composed of fat and is located at the innermost part of the skin layer. The dermis located above the subcutaneous tissue is composed of matrix proteins such as collagen and elastin, and includes blood vessels, nerves, and sweat glands. The epidermis located at the outermost part of the skin layer is occupied by keratinocytes.

Skin is gradually damaged by increasing age or by ultraviolet rays, external pollutants and stress, and the function of protecting the skin from these factors is weakened, resulting in skin dryness, wrinkles, itchiness, and inflammation due to bacterial infection. Among them, the stratum corneum, which is located on the outermost part of the epidermis, is composed of keratin and lipids, suppresses the penetration of harmful substances and evaporation of water, and protects the human body from bacteria and external stimuli.

When the skin is damaged, the proliferation of various cells is promoted following the inflammatory reaction, and these processes last from 2 days to 3 weeks after the skin is damaged. During this period, fibroblast proliferation and collagen production are promoted to fill the gaps in the damaged skin, and new cells and keratin components are reconstituted at the wound site.

Looking closely at the wound healing process of the skin, when the skin is damaged, the mast cells in the dermis secrete histamine, the blood vessels expand, and the inflammatory cells gather into the enlarged blood vessels. Among the gathered inflammatory cells, when white blood cells secrete cytokines, fibroblasts that maintain the elasticity of the dermis are activated to promote the secretion of substances such as collagen and mucopolysaccharide. Thereafter, the epidermis torn by the wound activates genes involved in cell proliferation and migration, and as a result, the epidermis is regenerated.

A representative substance that regulates the skin wound healing process is a growth factor, and this growth factor regulates the proliferation and differentiation of cells throughout the wound healing process. Growth factors include epidermal growth factor (EGF), plateletderived growth factor (PDGF), fibroblast growth factor (FGF), keratinocyte growth factor (KGF), transforming growth factor-β (TGF-β), and granulocyte colony stimulating factor (G-CSF). ) And VEGF (vascular endothelial growth factor), among which EGF (epidermal growth factor) plays an important role in the wound healing process.

Therefore, it is important not only to quickly heal the wound in the process of treating damage or wounds on the skin, but also to induce skin regeneration without an inflammatory reaction to treat without side effects or scars. Most of the ingredients for skin regeneration and wound healing that have been developed so far contain a large amount of artificially prepared compound ingredients, so that side effects that irritate the skin appear or there is a limit in skin regeneration ability.

Therefore, there is a need for research and commercialization of natural materials that have excellent effects on skin regeneration and wound healing and have no side effects.

On the other hand, kiwi fruit (Actinidia chinensis) is a deciduous vine plant of the Asteraceae family, and is a fruit variety made by crossing the Asteraceae plants. Originally, it was a plant grown in southern China, but it began to be cultivated mainly in New Zealand from the beginning of the 20th century. Blue daisies are known to defend neurotoxicity, inhibit the proliferation of gastric and lung cancer cells, and have anti-inflammatory and whitening effects. In addition, it contains 6 times more abundant tocopherol than apples, preventing damage to cell membranes early and delaying skin aging.

As a prior art related to red snail, Korean Patent No. 10-1181998 discloses a skin whitening composition using red snail leaf extract, and Korean Patent No. 10-1368293 discloses an anti-inflammatory composition and skin whitening agent composition using fermented red snail fruit. Korean Patent Registration No. 10-1081910 discloses a cosmetic composition containing kiwi extract that improves skin tone and prevents skin aging, but has not disclosed the effect of regenerating skin or wound healing effect of the extract of Astragalus chinensis. .

Accordingly, the inventors of the present invention that the extract of red snail stem extract can be usefully used as a cosmetic composition and quasi-drug composition for skin regeneration or wound healing by confirming the excellent effect of proliferation and migration of skin keratinocytes, promotion of collagen production, and anti-inflammatory effect. By confirming, the present invention was completed.

KR 10-1181998 B1 KR 10-1368293 B1 KR 10-1081910 B1

Accordingly, it is an object of the present invention to provide a cosmetic composition or a quasi-drug composition having an effect of promoting skin keratinocyte proliferation and migration, comprising a red snail stem extract.

Another object of the present invention is to provide a cosmetic composition or a quasi-drug composition having an effect of promoting collagen production, including a stem extract of bluetail.

Another object of the present invention is to provide a cosmetic composition or a quasi-drug composition having an anti-inflammatory effect comprising a bluetail stem extract.

A cosmetic composition for skin regeneration or wound healing containing the red snail stem extract of the present invention for achieving the above object as an active ingredient is characterized in that it contains the red snail stem extract as an active ingredient.

The bluetail stem extract is characterized in that it is contained in a concentration of 1ppm ~ 10,000ppm.

The snail stem extract is obtained by pulverizing the snail stalk, adding an extraction solvent of 1 to 50 times the amount of the dried product and stirring at 4 to 30°C for 2 to 7 days to extract the active ingredient, and the extraction solvent is water, organic It is a solvent or a mixed solvent thereof, and the organic solvent is characterized in that it is at least one selected from the group consisting of alcohols having 1 to 5 carbon atoms.

In addition, a pharmaceutical composition for external use for skin containing the extract of the blue sputum stem extract of the present invention as an active ingredient is characterized in that it comprises the extract of the blue sputum stem as an active ingredient.

The bluetail stem extract is characterized in that it is contained in a concentration of 1ppm ~ 10,000ppm.

According to the composition for skin regeneration or wound healing, which contains the extract of Satin stem extract according to the present invention as an active ingredient, a cosmetic composition or quasi-drug composition derived from a natural product has fewer side effects, has an effect of promoting skin keratinocyte proliferation and migration, and promoting collagen production. It has the advantage of excellent effect and anti-inflammatory effect.

1 is a photograph showing the results of testing the skin regeneration and wound healing efficacy of Experimental Example 1 according to the present invention.
Figure 2 is a graph showing the results of testing the cell proliferation promoting efficacy of Experimental Example 2 according to the present invention.
3 is a graph showing the results of experimentation of the collagen production promoting efficacy of Experimental Example 3 according to the present invention.
Figure 4 is a graph showing the results of testing the anti-inflammatory efficacy of Experimental Example 4 according to the present invention.
5 is a graph showing the results of testing the antioxidant efficacy of Experimental Example 5 according to the present invention.

Hereinafter, the present invention will be described in detail.

The composition according to the present invention has the effect of regenerating the skin and healing wounds, and is characterized in that it comprises a blue snail stem extract as an active ingredient. The extract of the blue snail stem has an effect of promoting skin regeneration and wound healing, as well as promoting collagen production and anti-inflammatory. In addition, since the extract of the bluefin chinensis does not irritate the skin and has excellent stability of the product after formulation, it is suitable as a cosmetic composition derived from a natural product and a pharmaceutical composition for external use of the skin.

At this time, the extract of the blue snail stem extract may be included in a concentration of 1 ppm to 10,000 ppm, more preferably 10 to 1,000 ppm in a cosmetic composition or a pharmaceutical composition for external skin, and if the concentration is too small, the effect of skin regeneration and wound healing It is insignificant, and even if the concentration is higher, there are disadvantages such as an increase in manufacturing cost as no additional effect does not appear.

In addition, it is sufficient if the extract of the bluetail stem is extracted by an extraction method well known in the art, and may be in a liquid state, but after extraction, a powdery product prepared through an additional process such as concentration under reduced pressure, freeze drying, or spray drying can be used. It is natural that it may be possible, and it is natural that such concentration, drying, and the like may be performed through various known methods.

A more detailed look at the manufacturing method of the bluetail stem extract is as follows.

The extract of the bluetail stem may be extracted and separated from nature using a method known in the art for extraction and separation.

At this time, the extraction is extracted from the stem of the bluetail using an appropriate solvent, and may include hot water extract, polar solvent-soluble extract, or non-polar solvent-soluble extract.

In addition, as a suitable solvent for extracting the extract from the bluetail stalk, any solvent allowed in the art may be used, but preferably, water, an organic solvent, or a mixture thereof may be used. Illustratively, various solvents such as an alcohol having 1 to 5 carbon atoms including purified water, methanol, ethanol, propanol, isopropanol, butanol, etc., alone or in two It may be mixed and used, but there is no need to be limited thereto.

And as the extraction method, any one of methods such as hot water extraction, cold precipitation extraction, reflux cooling extraction, solvent extraction, steam distillation method, ultrasonic extraction method, elution method, compression method, etc. can be selected and used, but there is no need to be limited thereto.

That is, there is no limitation on the method for preparing the blue stilt stem extract of the present invention, and any known method may be used.

As a specific example of the present invention, a stalk of bluetail is pulverized, and one or more extraction solvents selected from 1 to 50 volumes of water and an alcohol having 1 to 5 carbon atoms are added to the dried product, and 2 to 7 at 4 to 30°C. After the active ingredient is extracted by stirring for a day, it can be filtered and concentrated with a reduced pressure concentrator to prepare it. In addition, if necessary, it can be freeze-dried to prepare a powder. At this time, although the blue stilt stem may be used as it is, it is common to dry and use, and the drying method may use a natural drying method such as shade, but is not limited thereto. In addition, the pulverized particle size is also sufficient to be easily extracted, for example, about 10 to 100 mesh, but is not limited thereto.

The composition containing the bluetail stem extract according to the present invention may be provided as a cosmetic composition. The cosmetic composition may contain, without limitation, ingredients that are generally accepted in addition to the active ingredient, that is, a blue snail stem extract, such as diluents, excipients, buffers, dispersants, surfactants, colorants, flavors or sweeteners. Can include.

In addition, the cosmetic composition may further include functional cosmetic ingredients, moisturizers, and other additives that impart cosmetic functionality to the skin, and may additionally include one or more cosmetically acceptable carriers formulated in general skin cosmetics. . As such carrier components, animal oil, vegetable oil, wax, paraffin, silicone, lactose, silica, talc, water, ethanol, benzyl alcohol, glycerol aliphatic ester, aliphatic alcohol sulfate, etc. can be used, but the type is not limited. Is of course.

In addition, the cosmetic composition according to the present invention is selected from the group consisting of solutions, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, surfactant-containing cleansing oils, powder foundations, emulsion foundations, wax foundations, and sprays. It may have any one formulation, but is not limited thereto. At this time, the formulation example is the same as the preparation of a conventional cosmetic composition, except that the extract of the blue snail stem of the present invention is added.

In addition, the composition containing the extract of the blue snail stem of the present invention may be provided as a pharmaceutical composition for external use of the skin. The external pharmaceutical composition for skin may include, without limitation, pharmaceutically acceptable ingredients in addition to the active ingredient, that is, blue snail stem extract, and may include, for example, a pharmaceutically acceptable carrier, excipient, or diluent. The pharmaceutically acceptable carrier, excipient, or diluent is not limited as long as it does not impair the effect of the present invention, and for example, one of a filler, extender, binder, wetting agent, disintegrant, surfactant, lubricant, sweetener, fragrance, and preservative It may contain more than one species. Representative examples of the pharmaceutically acceptable carrier, excipient or diluent of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, maltitol, starch, gelatin, glycerin, gum acacia, alginate, calcium phosphate, calcium Carbonate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, propylene glycol, polyethylene glycol, vegetable oil , Injectable esters, Witepsol, Macrogol, Tween 61, cacao butter, and laurage.

In addition, the external pharmaceutical composition for skin may further contain one or more active ingredients exhibiting the same or similar function. For example, known wound healing or skin regeneration components may be included. Additional components may be included in an amount of 0.0001 to 10% by weight based on the total weight of the composition.

The pharmaceutical composition for external use for skin of the present invention may exemplify a disinfectant cleaner, a shower foam, an ointment, a wet tissue, a coating agent, etc., but is not limited thereto. It may be appropriately selected from conventional techniques known in the art.

In addition, a cosmetic composition or a pharmaceutical composition for external use of skin comprising the extract of the blue snail stem of the present invention can be used in a method of skin regeneration, including applying to the skin of an individual. The subject includes, without limitation, mammals including livestock, humans, and the like.

Hereinafter, the present invention will be described in more detail through the following experimental examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.

Example One: Bluefin Preparation of stem extract

After cleansing the stems of bluetail, dried in the shade, it was pulverized to about 100 mesh using a grinder. Thereafter, 200 ml of 70% ethanol was added to 20 g of the pulverized snail stem, extracted by stirring at 20° C. for 72 hours, filtered through a filter paper, concentrated under reduced pressure with a rotary concentrator, and freeze-dried to prepare a powdery snail stem extract.

Experimental example 1: Test to promote skin regeneration and wound healing

The following experiment was conducted to confirm the effect of promoting skin regeneration and wound healing of the bluetail stalk extract according to the present invention.

Specifically, using DMEM (10% FBS, 1% Penicillin/streptomysin) as a medium, HaCaT cells were inoculated in a 24 well plate at 2.0×10 ⁵ per well, and the medium was removed after 24 hours. Serum-free DMEM medium was treated with 1 ml per well, and cells were scraped using SPLScar™ (SPL Life Science, Korea). Then, the extract of the blue-tailed snail stem obtained in Example 1 was treated in the medium. Next, cell migration was measured after 72 hours incubation at 37°C and CO _{2 incubator.}

And the results are shown in FIG. 1. At this time, EGF (epidermal growth factor) was used as a positive control, and centella asiatica extract was used as a comparative example.

As can be seen in the accompanying Figure 1, as a result of treatment with the extract of Example 1 according to the present invention, it was found that it is superior to the centella asiatica extract as a comparative example, and a cell migration promoting effect at a level similar to that of the positive control EGF.

Therefore, it was found that the extract of the bluetail stem according to the present invention is excellent in promoting skin regeneration and wound healing.

Experimental example 2: Skin cells Proliferation promotion test

The following experiment was conducted in order to confirm the effect of promoting skin cell proliferation of the blue snail stem extract according to the present invention.

Specifically, using DMEM (10% FBS, 1% Penicillin/ streptomysin) as a medium, HaCaT cells were inoculated to a 96 well plate at 1.0×10 ⁴ per well, and the medium was removed after 24 hours. After the serum-free DMEM medium was treated with 100 µl per well, the extract of the blue snail stem obtained in Example 1 was treated in the medium by concentration. After culturing at 37°C _{for 24 hours in a CO 2} incubator, the medium was removed, and 100µl of MTT reagent was treated for each well, followed by _{incubation at 37°C for 3 hours in a CO 2} incubator to measure optical density at 570nm.

And the results are shown in FIG. 2. At this time, EGF (epidermal growth factor) was used as a positive control, and centella asiatica extract was used as a comparative example.

As can be seen in the accompanying Figure 2, as a result of treating the extract of Example 1 according to the present invention, it was confirmed that the proliferation of skin cells increased by about 13% at a concentration of 10 ppm. Therefore, it was found that the extract of the bluefin chinensis according to the present invention has an excellent effect of promoting skin cell proliferation.

Experimental example 3: collagen production promotion test

The following experiment was conducted to confirm the effect of promoting collagen production of fibroblasts of the blue-tailed snail stem extract according to the present invention.

Specifically, DMEM (10% FBS, 1% Penicillin/streptomysin) was used as a medium, and NIH3T3 cells were inoculated to a 96 well plate at 5.0×10 ⁴ per well. When 24 hours elapsed after the inoculation, the medium was changed to a serum-free DMEM medium, and the extract of the blue snail stem obtained in Example 1 was treated in the medium for each concentration. _{And after culturing in a CO 2} incubator at 37° C. for 24 hours, the amount of collagen production was measured at 555 nm by ELISA using a Sircol soluble collagen assay kit (Biocolor, UK).

And the results are shown in FIG. 3. Retinoic acid was used as a positive control, and centella asiatica extract was used as a comparative example.

As can be seen in the accompanying Figure 3, as a result of treatment of the extract of Example 1 according to the present invention, the collagen production promoting effect at a concentration of 20ppm increased by about 26%, the collagen production promoting effect than the comparative example Centella asiatica extract It was able to confirm the excellence. Therefore, it was found that the extract of the blue-tailed tuna stem according to the present invention has an excellent effect of promoting collagen production.

Experimental example 4: Inflammatory reaction inhibition test

The following experiment was conducted to confirm the inhibitory effect of the inflammatory response of the blue snail stem extract according to the present invention.

Specifically, DMEM (10% FBS, 1% Penicillin/streptomysin) was used as a medium, and Raw264.7 cells were inoculated to a 96 well plate at 2.0×10 ⁴ per well, and after 24 hours, the medium was After removal, LPS, which is an inflammation inducing factor, was added to a concentration of 1 ug/ml, and the extract of the blue snail stem obtained in Example 1 was treated in the medium by concentration. _{Then, after incubation in a CO 2} incubator at 37°C for 24 hours, an equal amount of Griss reagent (Promega, USA) was mixed with 50 µl of medium to measure the amount of NO (Nitric oxide) produced, and left for 20 minutes at room temperature in dark conditions. The absorbance (Optical Density) was measured at 540nm using.

And the results are shown in FIG. 4. L-NMMA was used as a positive control, and centella asiatica extract was used as a comparative example.

As can be seen in the accompanying Figure 4, as a result of treating the extract of Example 1 according to the present invention, the production of NO (Nitric oxide) induced by LPS at a concentration of 20 ppm was reduced by about 25%, compared to centella extract. It was confirmed that there is an excellent inhibitory effect on NO generation. Therefore, it was found that the extract of the bluetail stalk according to the present invention has excellent anti-inflammatory effects.

Experimental example 5: antioxidant test

The following experiment was conducted to confirm the antioxidant efficacy of the bluetail stem extract according to the present invention.

Specifically, a DPPH solution having a concentration of 0.32mM was added to the concentration-specific solution of the extract of the blue stilt obtained in Example 1, reacted for 20 minutes at room temperature in a light-shielding state, and then measured the optical density at 517 nm using ELISA. It was measured.

And the results are shown in FIG. 5. Ascorbic acid was used as a positive control group, and centella ascorbic acid was used as a comparative example.

As can be seen in the accompanying Figure 5, as a result of treatment of the extract of Example 1 according to the present invention, DPPH radical scavenging ability is increased by about 59% at 50 ppm, and DPPH radical scavenging ability at a level similar to that of the positive control can be confirmed. there was. Therefore, it was found that the extract of the bluetail stem according to the present invention has excellent antioxidant efficacy.

Experimental example 6: Cytotoxicity test

The following experiment was conducted to confirm the toxicity to skin cells of the blue snail stem extract according to the present invention.

Specifically, using DMEM (10% FBS, 1% Penicillin/streptomysin) as a medium, HaCaT cells were inoculated to a 96 well plate at 1×10 ⁴ per well, and after 24 hours, Example 1 The extract of the blue-tailed stalk obtained in was treated in the medium for each concentration. After culturing at 37°C _{for 72 hours in a CO 2} incubator, the medium was removed, 100µl of MTT reagent was treated for each well, and _{incubated at 37°C for 3 hours in a CO 2} incubator, and optical density was measured at 570nm.

As a result of treating the extract of Example 1 according to the present invention, it was confirmed that there was no direct effect on skin cell death up to a concentration of 100 ppm.

Experimental example 7: Raw material stability test

The following experiment was conducted to confirm the stability of the raw material of the blue snail stem extract according to the present invention.

Specifically, the extract obtained in Example 1 was prepared in a solution of 1,000 ppm concentration and under dark conditions, 25°C, 42°C and cycle (0°C→25°C→40°C, 8 hours each) conditions. After storage for 3 months, properties, pH changes and microbial contamination were measured.

As a result, it could be confirmed that the extract of the snail stem of Example 1 was stable without aggregation, precipitation, discoloration, discoloration, microbial contamination, and pH change under all storage conditions for 3 months. Therefore, it was confirmed that the stability of the bluetail stem extract according to the present invention is high.

As the specific parts of the present invention have been described in detail above, it is clear that these specific techniques are only preferred embodiments for those of ordinary skill in the art, and the scope of the present invention is not limited thereto. Therefore, it will be said that the practical scope of the present invention is defined by the appended claims and their equivalents.

Claims (5)

A cosmetic composition for skin regeneration, characterized in that it contains a bluetail stem extract as an active ingredient.
The method of claim 1,
The bluetail stem extract,
Cosmetic composition for skin regeneration, characterized in that contained in a concentration of 1ppm ~ 10,000ppm.
The method according to claim 1 or 2,
The bluetail stem extract,
The active ingredient was extracted by pulverizing the stalks of the bluetail, adding an extraction solvent of 1 to 50 times the amount of the dried product, and stirring at 4 to 30°C for 2 to 7 days,
The extraction solvent is water, an organic solvent, or a mixed solvent thereof,
The organic solvent is a cosmetic composition for skin regeneration, characterized in that at least one selected from the group consisting of alcohols having 1 to 5 carbon atoms.
A pharmaceutical composition for skin regeneration or wound healing for external use, characterized in that it comprises a blue-tailed tuna stem extract as an active ingredient.
The method of claim 4,
The bluetail stem extract,
An external pharmaceutical composition for skin regeneration or wound healing, characterized in that it is contained in a concentration of 1ppm to 10,000ppm.

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