KR102222919B1 - composition for preventing or treating of wound comprising Indirubin derivative compound - Google Patents

Abstract

The present invention relates to a composition for preventing or treating wounds, wherein the composition according to the present invention further comprises any one or more selected from the group consisting of indirubin derivatives alone or indirubin derivatives or extracts of Shiva, methyl vanillate, hesperidin, and quercitrin. By using the containing complex as an active ingredient, it can be usefully used as a pharmaceutical composition that can promote wound recovery and improve scars, and can be used in cosmetics for wound healing or improvement exerted from the function of promoting wound healing, including wound healing. I can.

Description

Pharmaceutical composition for preventing or treating wounds comprising an indirubin derivative as an active ingredient {composition for preventing or treating of wound comprising Indirubin derivative compound}

The present invention relates to a composition for preventing or treating wounds, and more particularly, to a pharmaceutical composition for preventing or treating wounds comprising an indirubin derivative as an active ingredient, and a cosmetic composition and food composition for treating or improving wounds comprising the same .

The skin acts as a barrier to protect the body, and it can be said to be the body's first line of defense, such as preventing the invasion of microorganisms. Wound healing and healing is the basis for restoration of tissue integrity and function after wounds, burns, abrasions and other traumatic injuries to the skin or surgical procedures. If wound healing is delayed or surgical wound opening occurs, significant clinical problems arise.

Currently, the wound treatments mainly used in Korea include Dongkuk Pharmaceutical's'Complex Madekasol' (released in 1985) and Dongwha Pharm's'Fucidine' (launched in 1980). Although the efficacy of the two agents is generally similar, fucidin has a little more strength in antibacterial activity, and madekasol has a little more strength in reducing scars.

Existing steroid ointments are effective for anti-inflammatory, immunosuppressive, and allergic diseases, but as side effects, they cause gombo marks, skin wrinkles, and folliculitis, and are not effective for bacterial diseases such as athlete's foot. In addition, ointments containing antibiotics can be resistant and can cause various side effects when used on soft skin wounds such as children. Therefore, in order to overcome the limitations of steroid ointments and antibiotic ointments, wound treatment drugs based on wound healing mechanisms are being introduced in the domestic market. In addition, accidental injuries that are more serious than normal skin wounds, large wounds that occur after surgery for skin diseases such as melanoma, and wounds associated with other diseases such as diabetes take time to heal and often cause scars. It's becoming a problem. Daewoong Pharmaceutical launched the first general medicine in Korea,'Ef Saesal Ointment', a wound treatment containing epidermal growth factor (EGF). Epithelial cell growth factor covers the skin of the wound (re-epithelialization) and promotes the creation of new flesh (granulation tissue proliferation), helping to prevent scarring. However, as epithelial cell growth factor is a protein, the cost and effort required for production are very high.

In order to overcome these limitations, stem cells have been proposed as a wound treatment material, but if the stem cells themselves are used, there is a possibility of carcinogenesis by DNA transfer, and due to the large size of the stem cells themselves, blood vessel occlusion or myocardial infarction can be caused. In addition, when transplanting using allogeneic cells such as umbilical cord blood, there is a problem that a rejection reaction occurs due to cell surface antigens. In addition, cell therapy products such as stem cells have disadvantages in that the manufacturing process is difficult and there are many restrictions on storage and transportation. Thus, while solving the fundamental problems of cell therapy products, there is a demand for a method capable of having the greatest therapeutic effect with minimal side effects.

Accordingly, the present inventors have endeavored to develop a wound treatment agent that has no side effects on the human body, but generally has a mild, continuous, and highly usable action. As a result, it prevents the infection of microorganisms in the wound area, and in addition to the wound recovery, scars (re-epithelialization And it is highly desirable to provide a novel composition for the healing of wounds and wounds that prevents granulation tissue proliferation).

Patent Document 1. Korean Registered Patent No. 10-1293762

The present invention has been conceived to solve the above problems, and an object of the present invention is to provide a pharmaceutical composition for preventing or treating wounds having an excellent wound healing effect, including an indirubin derivative as an active ingredient, and has no side effects on the human body. I want to provide.

Another object of the present invention is to provide a cosmetic composition or food composition for treating or improving wounds comprising the indirubin derivative as an active ingredient.

In addition, another object of the present invention is to provide a composition for skin regeneration and elasticity improvement comprising the indirubin derivative as an active ingredient.

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing or treating wounds comprising an indirubin derivative represented by the following Formula 1 or Formula 2 as an active ingredient.

[Formula 1]

[Formula 2]

The composition may further include an extract of Schizophrenia, or may further include any one or more selected from the group consisting of methyl vanylate, hesperidin, and quercitrin.

The wound may be injury due to burns, ulcers, trauma, post-surgical surgery, childbirth, chronic woumd, diabetic foot ulcers, or dermatitis. The wound may be applied to a wound after an implant or a patch that is a medical device.

The wound prevention or treatment may be due to promoting the movement speed of keratinocytes, activating the movement and proliferation of fibroblasts, and promoting collagen production.

The pharmaceutical composition may be for topical application to the skin.

The pharmaceutical composition may be formulated in any one formulation selected from the group consisting of creams, gels, ointments, emulsions, suspensions, sprays, and transdermal delivery pitch agents.

The composition may be an emulsion formulation further comprising an oil, a surfactant, and polyethylene glycol.

The mixing weight ratio of the oil, surfactant, and polyethylene glycol may be 0.3-30:1:2-2.5.

The active ingredient may be one containing 1 to 20% by weight based on the total weight of the composition.

The surfactant is polyoxyethylene sorbitan monolaurate (Tween 20), polyoxyethylene sorbitan monopalmitate (Tween 40), polyoxyethylene sorbitan monostearate (polyoxyethylene sorbitan monostearate). , Tween 60) and polyoxyethylene sorbitan oleate (Tween 80).

The oil may be any one or more selected from the group consisting of polyethoxylated castor oil (Kolliphor® EL), sunflower oil, and olive oil.

The composition may contain cyclodextrin for higher solubilization. Cyclodextrin may be 100 to 1000 parts by weight based on 100 parts by weight of the active ingredient present in the composition.

The emulsion formulation may be a stable formulation that does not show a change in wnt activity and solubility in distilled water under a temperature of 25 to 4°C when measured at 3 months.

The present invention provides a cosmetic composition for treating or improving wounds using an indirubin derivative represented by Formula 1 or Formula 2 as an active ingredient in order to achieve the above other object.

[Formula 1]

[Formula 2]

The composition may further include an extract of Schizophrenia, or may further include any one or more selected from the group consisting of methyl vanylate, hesperidin, and quercitrin.

The cosmetic composition may be any one selected from the group consisting of lotion, essence, lotion, cream, pack, gel, powder, foundation, and detergent.

The composition may be an emulsion formulation further comprising an oil, a surfactant, and polyethylene glycol.

The mixing weight ratio of the oil, surfactant, and polyethylene glycol may be 0.3-30:1:2-2.5.

The active ingredient may be one containing 1 to 20% by weight based on the total weight of the composition.

The surfactant is polyoxyethylene sorbitan monolaurate (Tween 20), polyoxyethylene sorbitan monopalmitate (Tween 40), polyoxyethylene sorbitan monostearate (polyoxyethylene sorbitan monostearate). , Tween 60) and polyoxyethylene sorbitan oleate (Tween 80).

The oil may be any one or more selected from the group consisting of polyethoxylated castor oil (Kolliphor® EL), sunflower oil, and olive oil.

The composition may contain cyclodextrin for higher solubilization. Cyclodextrin may be 100 to 1000 parts by weight based on 100 parts by weight of the active ingredient present in the composition.

The emulsion formulation may be a stable formulation that does not show a change in wnt activity and solubility in distilled water under a temperature of 25 to 4°C when measured at 3 months.

The present invention provides a food composition for treating or improving wounds comprising an indirubin derivative represented by the following Chemical Formula 1 or Chemical Formula 2 as an active ingredient in order to achieve the another object.

[Formula 1]

[Formula 2]

The composition may further include an extract of Schizophrenia, or may further include any one or more selected from the group consisting of methyl vanylate, hesperidin, and quercitrin.

The present invention provides a composition for skin regeneration and elasticity improvement comprising the indirubin derivative represented by Chemical Formula 1 or Chemical Formula 2 as an active ingredient in order to achieve the another object.

The composition may further include an extract of Schizophrenia, or may further include any one or more selected from the group consisting of methyl vanylate, hesperidin, and quercitrin.

The indirubin derivative may promote collagen synthesis, promote skin regeneration, and enhance skin elasticity.

The composition may be a formulation of a lotion, essence, lotion, cream, pack, gel, powder, foundation, or detergent.

The composition may be an emulsion formulation further comprising an oil, a surfactant, and polyethylene glycol.

The mixing weight ratio of the oil, surfactant, and polyethylene glycol may be 0.3-30:1:2-2.5.

The active ingredient may be one containing 1 to 20% by weight based on the total weight of the composition.

The surfactant is polyoxyethylene sorbitan monolaurate (Tween 20), polyoxyethylene sorbitan monopalmitate (Tween 40), polyoxyethylene sorbitan monostearate (polyoxyethylene sorbitan monostearate). , Tween 60) and polyoxyethylene sorbitan oleate (Tween 80).

The oil may be any one or more selected from the group consisting of polyethoxylated castor oil (Kolliphor® EL), sunflower oil, and olive oil.

The composition may contain cyclodextrin for higher solubilization. Cyclodextrin may be 100 to 1000 parts by weight based on 100 parts by weight of the active ingredient present in the composition.

The emulsion formulation may be a stable formulation that does not show a change in wnt activity and solubility in distilled water under a temperature of 25 to 4°C when measured at 3 months.

The indirubin derivative according to the present invention can be usefully used as a pharmaceutical composition capable of promoting wound recovery and improving scars, and can be used in cosmetics for wound healing or improvement exerted from the function of promoting wound healing, including wound healing. have.

In addition, since the indirubin derivative of the present invention is a stable compound that has been found to have little toxicity to the human body, when using them as a mixture, unlike conventional steroid drugs, it does not exhibit side effects to the human body, and can be used in pharmaceuticals, cosmetics or health foods.

In addition, the indirubin derivative according to the present invention promotes skin regeneration from external damage and improves elasticity by promoting collagen synthesis in skin tissues, and can be used in medicines, cosmetics, or health foods.

In addition, the extract or methyl vanillate, hesperidin, and quercitrin according to the present invention have little toxicity, and when used in combination with an indirubin derivative, unlike conventional steroid drugs, they do not exhibit side effects to the human body.

In addition, the present invention is a composition in which the composition is a mixture of any one or more of Shiva extract, methyl vanylate, hesperidin, and quercitrin and an indirubin derivative in a certain ratio, compared to a single composition for wound recovery, healing, and elasticity improvement, etc. It has the advantage of showing a high effect.

1 is a photograph showing the mobility of cells in keratinocytes in order to confirm the mobility of cells according to the treatment of the indirubin derivatives prepared from Examples 1, 2, and Comparative Examples 1 to 7; At this time, the control refers to keratinocyte cells not treated with the indirubin derivative.
2 is a graph quantitatively showing the cell migration rate in keratinocyte cells in order to confirm the mobility of cells according to the treatment of the indirubin derivatives prepared in Examples 1, 2, and Comparative Examples 1 to 7.
3 shows the passage of time (day 0, 14) for the mouse acute wound tissue treated with the indirubin derivative of Examples 1 and 2, the indirubin derivative of Comparative Example 1, and EGF (positive control) according to the present invention. It shows the pictures taken according to (1st) and the staining results. In addition, in the lower part of FIG. 3 (collagen staining), a mouse acute wound tissue treated with the indirubin derivative of Example 1 and Example 2, the indirubin derivative of Comparative Example 1, and EGF (positive control) according to the present invention was treated with Masson's trichrome, The results of staining with picrosirius red are shown.
4 shows the passage of time (1, 3, and the indirubin derivative of Example 1 and Example 2, the indirubin derivative of Comparative Example 1, and the mouse acute wound tissue treated with EGF (positive control) according to the present invention. It is a graph showing the quantification of the degree of re-epithelialization measured according to days 5, 9, and 12).
5 is a graph measuring wound size (cm 2) on day 14 in mice treated with the indirubin derivative of Example 1 and Example 2, the indirubin derivative of Comparative Example 1, and EGF (positive control) according to the present invention. .
Figure 6a is the movement of cells according to the treatment of the indirubin derivative prepared from Example 2, the methyl vanylate of Example 3, the hesperidin of Example 4, the quercitrin of Example 5, and the complexes prepared from Examples 7 to 9 In order to confirm the function, this is a picture showing the ability of cells to move in keratinocytes.
Figure 6b is the movement of cells according to the treatment of the indirubin derivative prepared from Example 2, the methyl vanylate of Example 3, the hesperidin of Example 4, the quercitrin of Example 5, and the complexes prepared from Examples 7 to 9 In order to confirm the function, it is a graph that quantitatively shows the cell migration rate in keratinocyte cells.
Figure 7 is an Example 2 indirubin derivative (0.5 mM) according to the present invention, the methyl vanylate of Example 3 (2.5 mM, 5 mM), the composite prepared from Example 7, EGF (positive control 1) and PTD+ The photographs and staining results of the mouse acute wound tissue treated with VPA (positive control 2) are shown according to the passage of time (day 0, day 14). In addition, the lower left graph of FIG. 7 is an example 2 indirubin derivative (0.5 mM) according to the present invention, methyl vanylate (2.5 mM, 5 mM) of Example 3, a composite prepared from Example 7, EGF (positive control) 1) and PTD+VPA (positive control 2) is a graph showing the quantification of the degree of re-epithelialization measured over time (day 0, day 14) for the mouse acute wound tissue treated. The lower right graph of FIG. 7 shows the indirubin derivative (0.5 mM) of Example 2, the methyl vanylate of Example 3 (2.5 mM, 5 mM), the composite prepared from Example 7, EGF (positive control 1) according to the present invention. ) And PTD+VPA (positive control 2) in mice treated with the wound size (cm2) on the first and 14th days.
FIG. 8A shows cells that were not treated with anything as a control (con), which was treated with the extract (1 µg/ml) of Schisandra chinensis prepared in Example 6 alone (1 µg/ml of Shisauna), and VPA (Valpro) as a positive control. Iksan) 100 μM was treated with keratinocytes, respectively, and 18 hours later, the cell mobility was taken.
8B shows the indirubin derivatives of Example 1 (0.1 μM, 1 μM), the indirubin derivatives of Example 2 (0.1 μM, 1 μM), and the complexes of Examples 10 and 11 (0.1 μM for 1 μg/ml of Shiva extract) , 1 μM of each of the indirubin derivatives) were treated with keratinocytes, respectively, and 18 hours later, the mobility of the cells was taken.
Figure 8c is a graph showing the calculation of the cell migration rate from Figures 8a, b.
9A shows the indirubin derivative of Example 1 (0.5 mM), the indirubin derivative of Example 2 (0.5 mM), the complex of Examples 10 and 11, the control group (con), and the extract of Shiva extract of Example 6 according to the present invention ( Shinamu), positive control (EGF and VPA) treated mouse acute wound tissue, the photographs taken according to the time (day 0, day 12) and staining results are shown.
9B is a graph showing the measurement of the size (mm 2) of acute wounds in each group of mice measured on day 0 in FIG. 9A.
9C is a graph showing the measurement of the size (mm 2) of acute wounds in each group of mice measured on the 12th day in FIG. 9A.
10 is a photograph of an emulsion solution prepared by dissolving the indirubin derivative prepared in Example 2 of the present invention in the various oils or surfactants, respectively.
11A is a ternary phase diagram for an emulsion solution prepared with a surfactant in which Tween 80 and PEG 400 are mixed in a ratio of 2:1, and FIG. 11B is an interface in which Tween 80 and PEG 400 are mixed in a ratio of 1:1. It is a ternary phase diagram for an emulsion solution prepared as an active agent, and FIG. 11C is a ternary phase diagram for an emulsion solution prepared as a surfactant in which Tween 80 and PEG 400 are mixed in a ratio of 1:2. In the ternary phase diagram of FIG. 11, regions forming a stable emulsion are indicated by red dots.
12 is a graph showing changes in relative solubility% of emulsion formulations F8 and F10-12 at room temperature (25° C.).
13 is a graph showing changes in relative solubility% of emulsion formulations F8 and F10-12 at low temperature (4° C.).
14 is a graph showing changes in the relative wnt reporter activity% of emulsion formulations F8 and F10-12 at room temperature (25° C.).
15 is a graph showing changes in the relative wnt reporter activity% of F8, F10~12 emulsion formulations at low temperature (4°C).

Hereinafter, various aspects and various embodiments of the present invention will be described in more detail.

One aspect of the present invention relates to a pharmaceutical composition for preventing or treating wounds comprising an indirubin derivative represented by the following Formula 1 or Formula 2 as an active ingredient.

[Formula 1]

[Formula 2]

The indirubin derivative of the present invention is a substance that shows little toxicity by cells even if it is treated for a long period of time, and has excellent cell regeneration, wound healing, and collagen production effects, thereby preventing wounds or treating wounds. It can be seen that it can be applied to various fields such as cosmetics and cosmetics.

The indirubin derivatives include 5-methoxyl indirubin-3'-oxime, 5-methoxyl indirubin-3'-methoxime, indirubin-3'-oxime, 6-bromoindirubin-3'-oxime , 5,6-dichloroindirubin, 5,6-dichloroindirubin-3'-oxime, 5,6-dichloroindirubin-3'-methoxime, 5,6-dichloroindirubin-3'-propyloxime, 6-chloro-5-nitroindirubin, 6-chloro-5-nitroindirubin-3'-oxime, 6-chloroindirubin-3'-methoxime, 5-chloroindirubin-3'-methoxime, 5 -Bromoindirubin-3'-oxime, 5-bromoindirubin-3'-methoxime, 5-bromoindirubin-3'-ethyloxime, 5,6-dichloroindirubin-3'-oximepropyl Various indirubin derivatives such as oxime and 6-chloroindirubin-3'-benzyloxime exist, but it was confirmed that the indirubin derivative represented by Formula 1 or Formula 2 was confirmed to have a significant effect on wound healing and recovery ( Experimental Example 1).

On the other hand, treatment of the indirubin derivative of Formula 1 or 2 through an in vitro (in virto ) experiment improves the wound closure rate (%) compared to the case of treatment with other conventional indirubin derivatives, and the wound healing effect is also simple. It has a healing effect (1.5 times, 2.0 times) more than a synergistic effect, and it was confirmed that the wound is being healed the fastest.

That is, it was confirmed through an experiment that the indirubin derivative represented by Formula 1 or Formula 2 is the most preferable among conventional indirubin derivatives.

In addition, as a result of applying the indirubin derivative of Formula 1 or Formula 2 to the wound of the mouse, the acute wound in the mouse was re-epithelialized equal to or faster than the positive control EGF, and various kinds of collagen tissues were formed. It can be seen that the wound healing markers increase. Specifically, in the case of the indirubin derivative (A3334) of Formula 1, it exhibits a slightly better wound healing effect than the positive control EGF, but the epidermal layer of the skin is formed so that no traces such as dents or dents are left as the wound heals. It was confirmed that it is more effective in'scar improvement' that recovers evenly.

In addition, in the case of the indirubin derivative (A3051) of Formula 2, it was confirmed that it exhibits a wound recovery effect of 2 to 6 times or more at the same time as that of the positive control EGF, and through this, it can be seen that the wound recovers more quickly. In addition, it was confirmed that the epidermal layer of the skin recovered cleanly and uniformly so that no traces such as dents or dents were left as the wound healed than when treated with the positive control EGF. It was confirmed that it is also very effective in'improvement'.

In other words, it was confirmed that the indirubin derivative of Formula 1 or Formula 2, unlike conventional wound treatment agents or indirubin derivatives, has an unexpected and remarkable effect of not only healing wounds but also improving scars that induce uniform recovery of the epidermal layer of the skin.

Accordingly, the indirubin derivative represented by Formula 1 or Formula 2 according to the present invention can be used as a pharmaceutical composition and cosmetic composition having a wound healing effect.

The composition according to the present invention has an effect of not only maximizing cell motility but also promoting and accumulating collagen in keratinocytes and fibroblasts that play an important role in wound healing.

In particular, keratinocytes are a type of cells found in the outer skin cells, forming the base layer of the outer skin cells, and forming a keratinized mature cell layer on the surface of the skin. The outer skin layer (corneal stralum) is formed by this. Wound healing rate is affected by the proliferation and migration rate of keratinocytes among several factors, and it was found that the composition according to the present invention increases the rate of movement of keratinocytes, thus showing a useful effect on wound healing. I can. In addition, it can be seen that the composition according to the present invention has an excellent wound healing effect as described above, as well as a'scar improvement' effect that induces uniform recovery of the epidermal layer of the skin. Most wound treatments are limited to wound treatment, and scars created during the wound treatment process must be treated separately with scar treatment agents. If scar treatment drugs are used for wound treatment, secondary problems such as inflammation may occur. Considering that both and scar treatment cannot be performed at the same time, it would be a very remarkable effect to have both wound treatment and scar improvement effects at the same time.

In other words, the composition according to the present invention may include, as an active ingredient, an indirubin derivative represented by Formula 1 or Formula 2, which has a remarkable wound prevention or healing effect (about 2 times) among various indirubin derivatives in the related art (Experimental Example One). As described above, the composition has the advantage of rapidly re-epithelializing acute wounds, promoting the production of collagen for various types, activating the accumulation and contraction thereof, and exhibiting the effect of activating the migration and proliferation of fibroblasts. Etc. It has a very remarkable wound healing effect.

The present invention may be a complex further comprising the extract of Shiva, and may be a complex further comprising any one or more low-molecular compounds selected from the group consisting of methyl vanylate, hesperidin, and quercitrin.

First, in the case of a composite obtained by mixing a Schizophrenia extract and an indirubin derivative at a certain ratio, it exhibits an excellent effect on wound healing. Specifically, when the indirubin derivative and the extract were treated together in an in-virto experiment, the wound closure rate (%) was improved compared to the case where the indirubin derivative of Formula 1 or Formula 2 was treated alone, It was confirmed that the wound healing effect also has a healing effect (2 times) more than a simple synergistic effect.

In particular, when treated with only the extract of Schizophrenia alone and when treated with the indirubin derivative alone, the values were about 1.5 to 2 times higher when the complex of Example 10 was treated than the sum of each. That is, it was confirmed that the composition for treating wounds of the present invention has a remarkable effect than that of using the extract and indirubin derivatives respectively, which exceeds the predictable value simply summed up the values obtained from each composition (Experimental Example 5 ).

In addition, it can be seen from the animal experiments in Experimental Example 6 that the complex with the indirubin derivative has a remarkable effect than the Shimane extract or the indirubin derivative alone, so it is more preferable.

The extract may be made of stems, leaves, fruits, pulverized products thereof, and mixtures thereof as a raw material, and the raw material is water, an anhydrous or hydrated lower alcohol having 1-4 carbon atoms, and a mixed solvent of the lower alcohol and water , Acetone, ethyl acetate, butyl acetate, chloroform, and may be extracted using a solvent selected from the group consisting of 1,3-butylene glycol as an extraction solvent.

The present invention may be a composite further comprising any one or more low-molecular compounds selected from the group consisting of methyl vanylate, hesperidin and quercitrin, and in the case of a composite in which the low-molecular compound and indirubin derivative are mixed in a certain ratio, it is used for wound healing. It shows an excellent effect against. Specifically, when the indirubin derivative and any one or more selected from the group consisting of methyl vanylate, hesperidin, and quercitrin were treated together in an in vitro (in virto) experiment, the indirubin derivative of Formula 1 or Formula 2 was used alone. Compared to the treated case, the wound closure rate (%) was improved, the wound healing effect also had a healing effect (1.4 to 2.3 times) more than a simple synergistic effect, and it was confirmed that the wound healed 6 hours faster. In other words, it was confirmed through an experiment that it is most preferable to use an indirubin derivative together with any one or more selected from the group consisting of methyl vanylate, hesperidine and quercitrin.

The composite in which any one or more low-molecular compounds selected from the group consisting of methyl vanylate, hesperidine, and quercitrin and any one or more indirubin derivatives selected from those represented by Formula 1 or Formula 2 are mixed in the experimental examples described later. As described above, as a result of analyzing the cell mobility of the low-molecular compound alone or a complex in which any one or more indirubin derivatives selected from those represented by Formula 2 were mixed, it was confirmed that the cell mobility was restored in 18 hours. In particular, when the methyl vanylate and the indirubin derivative (A3051) were mixed, it was confirmed that the result was 5 times higher than that of the control group (cell) that was not treated with anything.

Any one or more low-molecular compounds selected from the group consisting of methyl vanylate, hesperidin, and quercitrin is a component contained in Shimane extract, Earth japonica extract, citrus fruit extract, chinensis extract, Yeoseongcho extract, and Eoseongcho extract, the extract , If any one or more stems, leaves, fruits, pulverized products thereof, and mixtures thereof selected from one or more stems, leaves, fruits, pulverized products thereof, and mixtures thereof selected from earth japonica, tangerine, skewer, and Eoseongcho are used as raw materials, and specifically, the raw material is water, carbon number 1- Anhydrous or water-containing lower alcohol of 4, a mixed solvent of the lower alcohol and water, acetone, ethyl acetate, butyl acetate, chloroform, and a solvent selected from the group consisting of 1,3-butylene glycol as an extraction solvent. I can.

The methyl vanylate is mainly present in the extract of Earthquake, and the hesperidin is present in the extract of Shimane tree and the extract of citrus, and the quercitrin may be mainly present in the extract of Yeokki, Pyeonchuk extract, and Eoseongcho extract.

The indirubin derivative used in the present invention can be provided not only as a free substance, but also as a pharmaceutically acceptable salt, pharmaceutically acceptable solvate, pharmaceutically acceptable polymorph, or pharmaceutically acceptable prodrug thereof. have. The salt of the indirubin derivative is not particularly limited as long as it can be blended in medicine or cosmetics. It includes an inorganic salt or an organic salt, and may be an acid salt or an alkaline salt. In particular, in the case of a cation salt, alkali metal salts such as sodium salts and potassium salts, alkaline earth metal salts such as calcium salts, magnesium salts and barium salts; Basic amino acid salts such as arginine and lysine; Ammonium salts such as ammonium salts and tricyclohexyl ammonium salts; And various alkanol amine salts such as monoethanolamine salt, diethanolamine salt, triethanolamine salt, monoisopropanolamine salt, diisopropanolamine salt, and triisopropanolamine. Preferably, the salt is an alkali metal salt, more preferably a tetrasodium salt.

The composition according to the present invention exhibits excellent effects on wound healing ability and scar improvement, and can heal cell damage when cell damage occurs, and can improve wound healing, skin recovery, and scar, so that skin wounds such as abrasions It can be usefully used in the treatment and prevention of cuts, bruises, cuts, lacerations and bites. Most preferably, it may be due to a skin wound.

In the present specification, the term "including as an active ingredient" means including an amount sufficient to treat wounds on the skin of the present invention.

In the present specification, the term'wound prevention or treatment' includes suppressing the development of skin damage, reducing skin damage, removing, reducing, alleviating and improving symptoms, and is meant to include lowering the likelihood of skin wounds occurring. .

The wound is, as described above, burns, ulcers, trauma, post-surgical surgery, plastic surgery, implants, childbirth, chronic woumd, or dermatitis of the skin's epidermis, dermis or subcutaneous tissue. Means damage caused by. This development product can be used as a medical device when mounted on a patch.

In addition, the burn is preferably a sun, chemical, radiation or heat burn, the ulcer is preferably a diabetic ulcer, the chronic wound is preferably a pressure sore or pressure ulcer, and the dermatitis is impetigo (impetigo ), intertrigo, folliculitis, and eczema is preferably any one selected from the group consisting of, but is not limited thereto.

In the composition, the indirubin derivative may be included in an amount of 0.001 to 80% by weight based on the total weight of the composition, and if it is less than 0.01% by weight, sufficient wound prevention or healing effect may not appear, and when it exceeds 80% by weight, it is excessively included. May cause skin irritation.

When the composition further includes a serrata extract, the mixing weight ratio of the indirubin derivative represented by Formula 1 or 2 and the extract is not particularly limited thereto, but the extract and the indirubin derivative are based on 10 dry parts by weight of the extract. It is preferable that the indirubin derivative is mixed in an amount of 5 to 15 parts by weight, and if it is out of the above range, the increase in wound healing effect may be insignificant.

In addition, when the composition further includes any one or more low-molecular compounds selected from the group consisting of methyl vanylate, hesperidin and quercitrin, the indirubin derivative represented by Formula 1 or 2 and the methyl vanylate, hesperidin and quercitrin. The mixing weight ratio of any one or more low-molecular compounds selected from the group consisting of is not particularly limited thereto, but the low-molecular compound and the indirubin derivative are mixed at a ratio of 1 to 20 moles of the indirubin derivative based on 1 mole of the low-molecular compound. It is preferable, and if it is out of the above range, the increase in wound healing effect may be insignificant.

The concentration in which the extract or the low-molecular compound is contained in the composition is not particularly limited as long as it is a general range in the art, but the extract or the low-molecular compound is 0.001 to 5% by weight or 0.001 in liquid weight based on the total weight of the composition. To 50% by weight, if less than 0.01% by weight, a sufficient wound healing effect may not appear, and if it exceeds 5% by weight (dry weight) or 50% by weight (liquid weight), skin irritation may be induced. have.

The composition for preventing or treating wounds of the present invention may be applied in a general formulation in the art, such as oral administration or injection, or topical administration by parenteral administration or application, and a suitable dosage of the composition of the present invention may be applied to the patient's condition and It may vary depending on the body weight, the severity of the disease, the route and duration of administration of the drug form, and may be appropriately selected by those skilled in the art. However, the most preferred may be parenteral administration or topical administration by application.

When the composition of the present invention is administered by oral or injection, the composition according to the present invention may further include suitable carriers, excipients, and diluents commonly used in the manufacture of pharmaceuticals. It can be formulated and used in the form of oral dosage forms such as tablets, capsules, suspensions, emulsions, syrups, and aerosols, external preparations, suppositories, and sterile injectable solutions.

Carriers, excipients and diluents that may be included in the composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oils.

In the case of formulation, it is prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants that are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and these solid preparations include at least one excipient in the composition of the present invention, such as starch, calcium carbonate, and sucrose. It is prepared by mixing (sucrose) or lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use include suspensions, liquid solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as humectants, sweeteners, fragrances, and preservatives may be included. . Preparations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin paper, glycerol gelatin, and the like may be used.

The preferred dosage of the composition of the present invention varies depending on the condition and weight of the patient, the degree of disease, the form of the drug, the route and duration of administration, but may be appropriately selected by those skilled in the art. However, for desirable effects, the composition of the present invention is preferably administered at 0.0001 to 10 g/kg per day, preferably 0.001 to 8 mg/kg. Administration may be administered once a day, or may be divided several times. The above dosage does not limit the scope of the present invention in any way. Preferably, it can be administered within the range of 0.1 mg to 10 mg, 0.25 mg to 5 mg, and 0.5 mg to 2 mg per day. For example, 0.125mg, 0.25mg, 0.5mg, 0.75mg, 1mg, 1.5mg, 2mg can be administered per day.

Furthermore, the composition of the present invention may be parenteral or topical, and in the case of topical administration, it is most preferable to administer it in a manner that is applied directly to the skin.

When the composition is formulated for topical application in the form of a liquid, cream, lotion, gel or aerosol, suitable conventional additives, for example, preservatives, solvents that aid in penetration of medicines, softeners in the case of ointments and creams, etc. It may include. The topical preparations may also contain commercially compatible carriers such as ethanol or oleyl alcohol for cream or ointment bases and lotions. The carrier may constitute about 1 to about 98% of the formulation, and more generally up to about 80% of the formulation.

The composition according to the present invention may have a formulation that can be administered by a method such as direct application or dispersion to the skin or wound, for example, a formulation of a cream, lotion, ointment, aerosol, gel, or pack. Methods for formulations or formulations suitable for each formulation are well known in the art. When preparing these preparations, those skilled in the art can appropriately select and use various blending ingredients used to prepare ordinary external preparations.

As such ingredients, in the case of ointments, creams, gels, lotions, etc., white petrolatum, yellow petrolatum, lanolin, bleached beeswax, cetanol, stearyl alcohol, stearic acid, hydrogenated oil, gelled hydrocarbons, polyethylene glycol, liquid paraffin , Squalane and the like, oleic acid, isopropyl myristic acid, glycerin triisooctanoate, crotamiton, diethyl sebacate, diisopropyl adipate, hexyl laurate, fatty acids, fatty acid esters, aliphatic alcohols, vegetable oils There are solvents and dissolution aids such as tocopherol derivatives, antioxidants such as L-ascorbic acid, dibutylhydroxytoluene and butylhydroxyanisole, preservatives such as parahydroxybenzoic acid esters, glycerin, propylene glycol, sodium hyaluronate Moisturizing agents such as polyoxyethylene derivatives, glycerin fatty acid esters, sucrose fatty acid esters, sorbitan fatty acid esters, propylene glycol fatty acid esters, surfactants such as lecithin, carboxyvinyl polymer, xanthan gum, carboxymethylcellulose, carboxymethylcellulose sodium salts, And thickeners such as hydroxypropyl cellulose and hydroxypropyl methyl cellulose.

In the case of aerosols, in addition to the above ingredients used in the preparation of ointments, creams, gels, suspensions, emulsions, solutions, lotions, etc., various stabilizers, buffers, flavoring agents, suspending agents, emulsifiers, fragrances, preservatives , A dissolution aid, and other suitable additives can be blended.

The formulation of the composition may preferably be an emulsion formulation further comprising an oil, a surfactant, and polyethylene glycol.

In the emulsion formulation, the mixing weight ratio of the oil, surfactant, and polyethylene glycol may be 0.3-30:1:2-2.5, more preferably 10-20:1:2-2.5. The composition of the emulsion formulation can be determined by a pseudo three-phase diagram created according to a conventional method of determining the composition of the emulsion system. Specifically, after thoroughly mixing the oil phase (polyethoxylated castor oil (Kolliphor® EL)) and a surfactant (for example a mixture of tween 80 and polyethylene glycol) in different weight ratios in a specific ratio range, A pseudo three-phase diagram can be created by adding water to each proportion of the oil and surfactant mixture and marking the points corresponding to the emulsion-forming areas. From the group consisting of the active ingredients (indirubin derivatives or indirubin derivatives and Schizophrenia extract or methyl vanylate, hesperidin and quercitrin) by determining the emulsion region from the prepared pseudo-3 phase diagram and selecting a specific composition among the compositions contained in this region. It can be determined by the composition of the emulsion formulation that dissolves any one or more of the selected composites).

If the composition is an emulsion formulation, the active ingredient (indirubin derivatives or indirubin derivatives, and a compound of any one or more selected from the group consisting of chinensis extract or methyl vanillate, hesperidin, and quercitrin) is the total weight of the composition. It is preferable to contain 1 to 20% by weight based on.

The surfactant is a Tween-based nonionic surfactant that acts as a co-solubilizer, and is used as an emulsifier or wetting agent for oral or parenteral preparations in pharmaceuticals, and is also used as an additive in cosmetics and foods. It is also used as a substance for suppressing p-glycoprotein in order to increase the bioavailability of drugs. Tween-based surfactants include polyoxyethylene sorbitan monolaurate (Tween 20), polyoxyethylene sorbitan monopalmitate (Tween 40), and polyoxyethylene sorbitan monolaurate (Tween 40). polyoxyethylene sorbitan monostearate, Tween 60) and polyoxyethylene sorbitan oleate (Tween 80). The tween-based surfactant is a stable substance approved for use by the US Food and Drug Administration for intravenous injection of the human body.

In addition, the polyethylene glycol is an amphoteric polymer having hydrophilicity and hydrophobicity, and the polyethylene glycol is a liquid when it has a low molecular weight, but becomes a solid as the molecular weight increases. The polyethylene glycol may be any one selected from the group consisting of PEG 150, 300, 400, 1000, 6000, 8000, 10000, 20000, 30000, and 40000. In this case, the PEG300 refers to polyethylene glycol having a molecular weight of 300. And, polyethylene glycol having a molecular weight exceeding 10,000 is also referred to as polyethylene oxide (PEO). Among these, PEG400 exists in liquid form and is frequently used for solubilization of various poorly soluble drugs, and is especially used for oral and parenteral (intravenous injection, subcutaneous injection, intramuscular injection, etc.) of the human body by the US Food and Drug Administration (FDA). It is a stable material that has been approved for.

The surfactant used in the examples of the present invention is polyoxyethylene sorbitan oleate (Tween 80), and polyethylene glycol is PEG 400, which has amphiphilic properties and is non-ionic that can be dispersible in water. , In a poorly soluble active ingredient (indirubin derivative represented by Chemical Formula 1 or 2 above; and an active ingredient comprising a complex of any one or more low-molecular compounds selected from the group consisting of S. tree extract or methyl vanylate, hesperidin, and quercitrin) It is possible to provide an optimized, emulsion formulation.

The oil may be any one or more selected from the group consisting of polyethoxylated castor oil (Kolliphor® EL), sunflower oil, and olive oil.

The composition may contain cyclodextrin for higher solubilization. Cyclodextrin may be 100 to 1000 parts by weight based on 100 parts by weight of the active ingredient present in the composition.

The emulsion formulation is a stable formulation that does not show changes in wnt activity and solubility in distilled water under conditions of 25 to 4°C when measured at 3 months. It enables the activity of the active ingredient to be maintained for a long period of time and effectively absorbs into the body. It was confirmed that it can be improved.

In addition, for preventing or treating wounds comprising any one or more selected from the group consisting of the indirubin derivative represented by the following formula (1), the indirubin derivative represented by the formula (2), serrata extract, methyl vanylate, hesperidin, and quercitrin The preparation method of the emulsion pharmaceutical composition is as follows.

1) preparing a first solution by mixing oil, surfactant and polyethylene glycol;

2) a second solution containing at least one active ingredient selected from the group consisting of the indirubin derivative represented by the formula 1, the indirubin derivative represented by the formula 2, the chinensis extract, methyl vanylate, hesperidin, and quercitrin Manufacturing steps;

3) preparing an emulsion composition by mixing a second solution with the first solution.

The second solution may contain cyclodextrin for higher solubilization.

The polyethylene glycol is an amphoteric polymer having hydrophilicity and hydrophobicity, and the polyethylene glycol is a liquid when it has a low molecular weight, but becomes a solid as the molecular weight increases. The polyethylene glycol may be any one selected from the group consisting of PEG 150, 300, 400, 1000, 6000, 8000, 10000, 20000, 30000, and 40000. In this case, the PEG300 refers to polyethylene glycol having a molecular weight of 300. And, polyethylene glycol having a molecular weight exceeding 10,000 is also referred to as polyethylene oxide (PEO). Among these, PEG400 exists in liquid form and is frequently used for solubilization of various poorly soluble drugs, and is especially used for oral and parenteral (intravenous injection, subcutaneous injection, intramuscular injection, etc.) of the human body by the US Food and Drug Administration (FDA). It is a stable material that has been approved for.

The surfactant is a Tween-based nonionic surfactant that acts as a co-solubilizer, and is used as an emulsifier or wetting agent for oral or parenteral preparations in pharmaceuticals, and is also used as an additive in cosmetics and foods. It is also used as a substance for suppressing p-glycoprotein in order to increase the bioavailability of drugs. Tween-based surfactants include polyoxyethylene sorbitan monolaurate (Tween 20), polyoxyethylene sorbitan monopalmitate (Tween 40), and polyoxyethylene sorbitan monolaurate (Tween 40). polyoxyethylene sorbitan monostearate, Tween 60) and polyoxyethylene sorbitan oleate (Tween 80). The tween-based surfactant is a stable substance approved for use by the US Food and Drug Administration for intravenous injection of the human body.

The mixing weight ratio of the oil, surfactant and polyethylene glycol may be 0.3-30:1:2-2.5, more preferably 10-20:1:2-2.5. If the composition is in an emulsion formulation, the active ingredient (a compound in which any one or more selected from the group consisting of indirubin derivatives or indirubin derivatives and serrata extract, methyl vanillate, hesperidin, and quercitrin is mixed) is the total composition of the composition. It is preferable to contain 1 to 20% by weight based on the weight.

When water was added to the emulsion composition of the present invention to take a microscopic picture and compared, the emulsion of the present invention was completely dissolved to form an emulsion formulation (F8) in a solution state. In addition, as a result of observing the stability of the emulsion of the present invention, the droplets have a spherical shape, the average diameter is 20 to 1500 nm, the preferred diameter is measured to be 30 to 50 nm to form nano-sized droplets, and a narrow range of size distribution Is shown.

In addition, it was confirmed that the emulsion of the present invention maintains stability for a long period of time because there is no change in solubility and activity of active ingredients for 3 months at room temperature. Furthermore, when the emulsion formulation of the present invention is administered orally or applied to a topical area, absorption can be effectively improved.

In addition, another aspect of the present invention relates to a cosmetic composition for treating or improving wounds using the indirubin derivative represented by Chemical Formula 1 or Chemical Formula 2 as an active ingredient.

[Formula 1]

[Formula 2]

The indirubin derivative of the present invention is a substance that shows little toxicity by cells even if it is treated for a long period of time, and by confirming that it exhibits excellent cell regeneration, wound healing, and collagen production effects, it provides a cosmetic effect through wound prevention or wound treatment. can do.

The indirubin derivatives include 5-methoxyl indirubin-3'-oxime, 5-methoxyl indirubin-3'-methoxime, indirubin-3'-oxime, 6-bromoindirubin-3'-oxime , 5,6-dichloroindirubin, 5,6-dichloroindirubin-3'-oxime, 5,6-dichloroindirubin-3'-methoxime, 5,6-dichloroindirubin-3'-propyloxime, 6-chloro-5-nitroindirubin, 6-chloro-5-nitroindirubin-3'-oxime, 6-chloroindirubin-3'-methoxime, 5-chloroindirubin-3'-methoxime, 5 -Bromoindirubin-3'-oxime, 5-bromoindirubin-3'-methoxime, 5-bromoindirubin-3'-ethyloxime, 5,6-dichloroindirubin-3'-oximepropyl Various indirubin derivatives such as oxime and 6-chloroindirubin-3'-benzyloxime exist, but it was confirmed that the indirubin derivative represented by Formula 1 or Formula 2 was confirmed to have a significant effect on wound healing and recovery ( Experimental Example 1).

On the other hand, treatment of the indirubin derivative of Formula 1 or 2 through an in vitro (in virto ) experiment improves the wound closure rate (%) compared to the case of treatment with other conventional indirubin derivatives, and the wound healing effect is also simple. It has a healing effect (1.5 times, 2.0 times) more than a synergistic effect, and it was confirmed that the wound is being healed the fastest. Therefore, it was confirmed through an experiment that the indirubin derivative of Formula 1 or Formula 2 is most preferable among conventional indirubin derivatives.

In addition, as a result of applying the indirubin derivative of Formula 1 or Formula 2 to the wound of the mouse, the acute wound in the mouse was re-epithelialized equal to or faster than the positive control EGF, and various kinds of collagen tissues were formed. It can be seen that the wound healing markers increase. Specifically, in the case of the indirubin derivative (A3334) of Formula 1, it exhibits a slightly better wound healing effect than the positive control EGF, but the epidermal layer of the skin is formed so that no traces such as dents or dents are left as the wound heals. It was confirmed that it is more effective in'scar improvement' that recovers evenly.

In addition, in the case of the indirubin derivative (A3051) of Formula 2, it was confirmed that it exhibits a wound recovery effect of 2 to 6 times or more at the same time as that of the positive control EGF, and through this, it can be seen that the wound recovers more quickly. In addition, it was confirmed that the epidermal layer of the skin recovered cleanly and uniformly so that no traces such as lumps or dents were left as the wound healed than when treated with EGF, the positive control. It was confirmed that it is also very effective in'improvement'

In other words, it was confirmed that the indirubin derivative of Formula 1 or Formula 2, unlike conventional wound treatment agents or indirubin derivatives, has an unexpected and remarkable effect of not only healing wounds but also improving scars that induce uniform recovery of the epidermal layer of the skin.

In the composition, the indirubin derivative may be included in an amount of 0.001 to 80% by weight based on the total weight of the composition, and if it is less than 0.01% by weight, sufficient wound prevention or healing effect may not appear, and when it exceeds 80% by weight, it is excessively included. May cause skin irritation.

The present invention may be a complex further comprising the extract of Shiva, and may be a complex further comprising any one or more low-molecular compounds selected from the group consisting of methyl vanylate, hesperidin, and quercitrin.

First, in the case of a composite obtained by mixing a Schizophrenia extract and an indirubin derivative at a certain ratio, it exhibits an excellent effect on wound healing. Specifically, when the indirubin derivative and the extract were treated together in an in-virto experiment, the wound closure rate (%) was improved compared to the case where the indirubin derivative of Formula 1 or Formula 2 was treated alone, It was confirmed that the wound healing effect also has a healing effect (2 times) more than a simple synergistic effect.

In particular, when treated with only the extract of Schizophrenia alone and when treated with the indirubin derivative alone, the values were about 1.5 to 2 times higher when the complex of Example 10 was treated than the sum of each. That is, it was confirmed that the composition for treating wounds of the present invention has a remarkable effect than that of using the extract and indirubin derivatives respectively, which exceeds the predictable value simply summed up the values obtained from each composition (Experimental Example 5 ).

In addition, it can be seen from the animal experiments in Experimental Example 6 that the complex with the indirubin derivative has a remarkable effect than the Shimane extract or the indirubin derivative alone, so it is more preferable.

The extract may be made of stems, leaves, fruits, pulverized products thereof, and mixtures thereof as a raw material, and the raw material is water, an anhydrous or hydrated lower alcohol having 1-4 carbon atoms, and a mixed solvent of the lower alcohol and water , Acetone, ethyl acetate, butyl acetate, chloroform, and may be extracted using a solvent selected from the group consisting of 1,3-butylene glycol as an extraction solvent.

Next, in the case of a composite obtained by mixing at least one low-molecular compound selected from the group consisting of methyl vanylate, hesperidin, and quercitrin and an indirubin derivative in a certain ratio, it exhibits excellent effects on wound healing. Specifically, when the indirubin derivative and any one or more selected from the group consisting of methyl vanylate, hesperidin, and quercitrin were treated together in an in vitro (in virto) experiment, the indirubin derivative of Formula 1 or Formula 2 was used alone. Compared to the case of treatment, the wound closure rate (%) was improved, the wound healing effect also had a healing effect (1.4 to 2.3 times) more than a simple synergistic effect, and it was confirmed that the wound was healed 6 hours faster. In other words, it was confirmed through an experiment that it is most preferable to use an indirubin derivative together with any one or more of any one or more selected from the group consisting of methyl vanylate, hesperidin, and quercitrin.

The composite in which any one or more selected from the group consisting of methyl vanylate, hesperidin, and quercitrin and any one or more indirubin derivatives selected from those represented by Formula 1 or Formula 2 are mixed, as in Experimental Examples to be described later. , Methyl vanylate, hesperidin, and quercitrin alone or a mixture of one or more indirubin derivatives selected from among those represented by the formula (2) as a result of analyzing the cell mobility, it was confirmed that the cell mobility was restored in 18 hours. . In particular, when the methyl vanylate and indirubin derivative (A3051) were mixed, it was confirmed that the result was 5 times higher than that of the control group (cell) that was not treated with anything.

Any one or more low-molecular compounds selected from the group consisting of methyl vanylate, hesperidin, and quercitrin is a component contained in Shimane extract, Earth japonica extract, citrus fruit extract, chinensis extract, Yeoseongcho extract, and Eoseongcho extract, the extract , If any one or more stems, leaves, fruits, pulverized products thereof, and mixtures thereof selected from one or more stems, leaves, fruits, pulverized products thereof, and mixtures thereof selected from earth japonica, tangerine, skewer, and Eoseongcho are used as raw materials, and specifically, the raw material is water, carbon number 1- Anhydrous or water-containing lower alcohol of 4, a mixed solvent of the lower alcohol and water, acetone, ethyl acetate, butyl acetate, chloroform, and a solvent selected from the group consisting of 1,3-butylene glycol as an extraction solvent. I can.

The methyl vanylate is mainly present in the extract of Earthquake, and the hesperidin is present in the extract of Shimane tree and the extract of citrus, and the quercitrin may be mainly present in the extract of Yeokki, Pyeonchuk extract, and Eoseongcho extract.

When the composition further includes a serrata extract, the mixing weight ratio of the indirubin derivative represented by Formula 1 or 2 and the extract is not particularly limited thereto, but the extract and the indirubin derivative are based on 10 dry parts by weight of the extract. It is preferable that the indirubin derivative is mixed in an amount of 5 to 15 parts by weight, and if it is out of the above range, the increase in wound healing effect may be insignificant.

When the composition further comprises any one or more selected from the group consisting of methyl vanylate, hesperidin and quercitrin, selected from the group consisting of the indirubin derivative represented by Formula 1 or 2 and the methyl vanylate, hesperidine and quercitrin The mixing weight ratio of any one or more of the low molecular weight compounds is not particularly limited thereto, but the low molecular weight compound and the indirubin derivative are preferably mixed in a 1 to 20 mole ratio of the indirubin derivative based on 1 mole of the low molecular compound, and the If it is out of range, the increase in wound healing effect may be insignificant.

The concentration in which the extract or the low-molecular compound is contained in the composition is not particularly limited as long as it is a general range in the art, but the extract or the low-molecular compound is 0.001 to 5% by weight or 0.001 in liquid weight based on the total weight of the composition. To 50% by weight, if less than 0.01% by weight, a sufficient wound healing effect may not appear, and if it exceeds 5% by weight (dry weight) or 50% by weight (liquid weight), skin irritation may be induced. have.

The cosmetic composition according to the present invention may further include an additive capable of increasing the cosmetic effect through wound improvement, and is not particularly limited thereto.

The present invention may be a complex further comprising the extract of Shiva, and may be a complex further comprising any one or more low-molecular compounds selected from the group consisting of methyl vanylate, hesperidin, and quercitrin.

The cosmetic composition according to the present invention is a lotion, essence. It can be prepared in lotion, cream, pack, gel, powder, foundation or detergent formulation, for example, lotion, nutrition lotion, nutrition essence, massage cream, beauty bath water additive, body lotion, body milk, bath oil, baby oil, Baby powder, shower gel, shower cream, sunscreen lotion, sunscreen cream, suntan cream, skin lotion, skin cream, sunscreen cosmetics, cleansing milk, depilatory makeup, face and body lotion, face and body cream, skin whitening Cream, hand lotion, hair lotion, cosmetic cream, jasmine oil, bath soap, water soap, beauty soap, shampoo, hand sanitizer (hand cleaner), medicinal soap, non-medical, cream soap, facial wash, body cleaner, scalp cleaner, hair It can be prepared in the form of rinse, cosmetic soap, tooth whitening gel, toothpaste, and the like. To this end, the composition of the present invention may further include a solvent or a suitable carrier, excipient, or diluent that is commonly used in the manufacture of a cosmetic composition.

The type of solvent that can be further added to the cosmetic composition of the present invention is not particularly limited, but, for example, water, saline, DMSO, or a combination thereof may be used, and as a carrier, excipient or diluent, purified water, oil, wax , Fatty acids, fatty alcohols, fatty acid esters, surfactants, humectants, thickeners, antioxidants, viscosity stabilizers, chelating agents, buffers, lower alcohols, and the like, but are not limited thereto. In addition, whitening agents, moisturizing agents, vitamins, sunscreen agents, perfumes, dyes, antibiotics, antibacterial agents, and antifungal agents may be included as needed.

Hydrogenated vegetable oil, castor oil, cottonseed oil, olive oil, palm seed oil, jojoba oil, and avocado oil may be used as the oil. Can be.

Stearic acid, linoleic acid, linolenic acid, and oleic acid may be used as the fatty acid, and cetyl alcohol, octyldodecanol, oleyl alcohol, panthenol, lanolin alcohol, stearyl alcohol, hexadecanol may be used as fatty alcohols. In addition, as the fatty acid ester, isopropyl myristate, isopropyl palmitate, and butyl stearate may be used. In addition, as the surfactant, cationic surfactants, anionic surfactants, and nonionic surfactants known in the art can be used, and surfactants derived from natural products are preferred as far as possible.

In addition, hygroscopic agents, thickeners, antioxidants, and the like, which are widely known in the cosmetic field may be included, and the types and amounts thereof are as known in the art.

The cosmetic composition of the present invention may preferably be an emulsion formulation further comprising an oil, a surfactant, and polyethylene glycol.

In the emulsion formulation, the mixing weight ratio of the oil, surfactant, and polyethylene glycol may be 0.3-30:1:2-2.5, more preferably 10-20:1:2-2.5. The composition of the emulsion formulation can be determined by a pseudo three-phase diagram created according to a conventional method of determining the composition of the emulsion system. Specifically, after thoroughly mixing the oil phase (polyethoxylated castor oil (Kolliphor® EL)) and a surfactant (for example a mixture of tween 80 and polyethylene glycol) in different weight ratios in a specific ratio range, A pseudo three-phase diagram can be created by adding water to each proportion of the oil and surfactant mixture and marking the points corresponding to the emulsion-forming areas. From the group consisting of the active ingredients (indirubin derivatives or indirubin derivatives and Schizophrenia extract, methyl vanillate, hesperidin and quercitrin) by determining the emulsion region from the created pseudo-3 phase diagram and selecting a specific composition among the compositions contained in this region. It can be determined by the composition of the emulsion formulation dissolving any one or more of the selected composites).

If the composition is in an emulsion formulation, the active ingredient (a mixture of indirubin derivatives or indirubin derivatives, and any one or more selected from the group consisting of Shiva extract, methyl vanillate, hesperidin, and quercitrin) is the total weight of the composition. It is preferable to contain 1 to 20% by weight based on.

The surfactant is a Tween-based nonionic surfactant that acts as a co-solubilizer, and is used as an emulsifier or wetting agent for oral or parenteral preparations in pharmaceuticals, and is also used as an additive in cosmetics and foods. It is also used as a substance for suppressing p-glycoprotein in order to increase the bioavailability of drugs. Tween-based surfactants include polyoxyethylene sorbitan monolaurate (Tween 20), polyoxyethylene sorbitan monopalmitate (Tween 40), and polyoxyethylene sorbitan monolaurate (Tween 40). polyoxyethylene sorbitan monostearate, Tween 60) and polyoxyethylene sorbitan oleate (Tween 80). The tween-based surfactant is a stable substance approved for use by the US Food and Drug Administration for intravenous injection of the human body.

In addition, the polyethylene glycol is an amphoteric polymer having hydrophilicity and hydrophobicity, and the polyethylene glycol is a liquid when it has a low molecular weight, but becomes a solid as the molecular weight increases. The polyethylene glycol may be any one selected from the group consisting of PEG 150, 300, 400, 1000, 6000, 8000, 10000, 20000, 30000, and 40000. In this case, the PEG300 refers to polyethylene glycol having a molecular weight of 300. And, polyethylene glycol having a molecular weight exceeding 10,000 is also referred to as polyethylene oxide (PEO). Among these, PEG400 exists in liquid form and is frequently used for solubilization of various poorly soluble drugs, and is especially used for oral and parenteral (intravenous injection, subcutaneous injection, intramuscular injection, etc.) of the human body by the US Food and Drug Administration (FDA). It is a stable material that has been approved for.

The surfactant used in the examples of the present invention is polyoxyethylene sorbitan oleate (Tween 80), and polyethylene glycol is PEG 400, which has amphiphilic properties and is non-ionic that can be dispersible in water. , Optimized for a poorly soluble active ingredient (an active ingredient comprising any one or more complexes selected from the group consisting of the indirubin derivative represented by Chemical Formula 1 or 2 or the extract of Schisandra chinensis, methyl vanylate, hesperidin, and quercitrin), Emulsion formulations can be provided.

The oil may be any one or more selected from the group consisting of polyethoxylated castor oil (Kolliphor® EL), sunflower oil, and olive oil.

The composition may contain cyclodextrin for higher solubilization. Cyclodextrin may be 100 to 1000 parts by weight based on 100 parts by weight of the active ingredient present in the composition.

The emulsion formulation is a stable formulation that does not show changes in wnt activity and solubility in distilled water under conditions of 25 to 4°C when measured at 3 months. It enables the activity of the active ingredient to be maintained for a long period of time and effectively absorbs into the body. It was confirmed that it can be improved.

In addition, for treating or improving wounds comprising any one or more selected from the group consisting of the indirubin derivative represented by the following Formula 1, the indirubin derivative represented by the Formula 2, the serrata extract, methyl vanylate, hesperidin, and quercitrin The preparation method of the emulsion cosmetic composition is as follows.

1) preparing a first solution by mixing oil, surfactant and polyethylene glycol;

2) a second solution containing at least one active ingredient selected from the group consisting of the indirubin derivative represented by the formula 1, the indirubin derivative represented by the formula 2, the chinensis extract, methyl vanylate, hesperidin, and quercitrin Manufacturing steps;

3) preparing an emulsion composition by mixing a second solution with the first solution.

The second solution may contain cyclodextrin for higher solubilization.

The polyethylene glycol is an amphoteric polymer having hydrophilicity and hydrophobicity, and the polyethylene glycol is a liquid when it has a low molecular weight, but becomes a solid as the molecular weight increases. The polyethylene glycol may be any one selected from the group consisting of PEG 150, 300, 400, 1000, 6000, 8000, 10000, 20000, 30000, and 40000. In this case, the PEG300 refers to polyethylene glycol having a molecular weight of 300. And, polyethylene glycol having a molecular weight exceeding 10,000 is also referred to as polyethylene oxide (PEO). Among these, PEG400 exists in liquid form and is frequently used for solubilization of various poorly soluble drugs, and is especially used for oral and parenteral (intravenous injection, subcutaneous injection, intramuscular injection, etc.) of the human body by the US Food and Drug Administration (FDA). It is a stable material that has been approved for.

The surfactant is a Tween-based nonionic surfactant that acts as a co-solubilizer, and is used as an emulsifier or wetting agent for oral or parenteral preparations in pharmaceuticals, and is also used as an additive in cosmetics and foods. It is also used as a substance for suppressing p-glycoprotein in order to increase the bioavailability of drugs. Tween-based surfactants include polyoxyethylene sorbitan monolaurate (Tween 20), polyoxyethylene sorbitan monopalmitate (Tween 40), and polyoxyethylene sorbitan monolaurate (Tween 40). polyoxyethylene sorbitan monostearate, Tween 60) and polyoxyethylene sorbitan oleate (Tween 80). The tween-based surfactant is a stable substance approved for use by the US Food and Drug Administration for intravenous injection of the human body.

The mixing weight ratio of the oil, surfactant and polyethylene glycol may be 0.3-30:1:2-2.5, more preferably 10-20:1:2-2.5. If the composition is in an emulsion formulation, the active ingredient (a compound in which any one or more selected from the group consisting of indirubin derivatives or indirubin derivatives and serrata extract, methyl vanillate, hesperidin, and quercitrin is mixed) is the total composition of the composition. It is preferable to contain 1 to 20% by weight based on the weight.

When water was added to the emulsion composition of the present invention to take a microscopic picture and compared, the emulsion of the present invention was completely dissolved to form an emulsion formulation (F8) in a solution state. In addition, as a result of observing the stability of the emulsion of the present invention, the droplets have a spherical shape, the average diameter is 20 to 1500 nm, the preferred diameter is measured to be 30 to 50 nm to form nano-sized droplets, and a narrow range of size distribution Is shown.

In addition, it was confirmed that the emulsion of the present invention maintains stability for a long period of time because there is no change in solubility and activity of active ingredients for 3 months at room temperature. Furthermore, when the emulsion formulation of the present invention is administered orally or applied to a topical area, absorption can be effectively improved.

In addition, another aspect of the present invention relates to a food composition for treating or improving wounds comprising an indirubin derivative represented by Formula 1 or Formula 2 as an active ingredient.

[Formula 1]

[Formula 2]

The indirubin derivative of the present invention is a substance that hardly exhibits toxicity by cells even if it is treated for a long period of time, and can impart wound healing or improvement effects by confirming that it exhibits excellent cell regeneration, wound healing, and collagen production effects.

The indirubin derivatives include 5-methoxyl indirubin-3'-oxime, 5-methoxyl indirubin-3'-methoxime, indirubin-3'-oxime, 6-bromoindirubin-3'-oxime , 5,6-dichloroindirubin, 5,6-dichloroindirubin-3'-oxime, 5,6-dichloroindirubin-3'-methoxime, 5,6-dichloroindirubin-3'-propyloxime, 6-chloro-5-nitroindirubin, 6-chloro-5-nitroindirubin-3'-oxime, 6-chloroindirubin-3'-methoxime, 5-chloroindirubin-3'-methoxime, 5 -Bromoindirubin-3'-oxime, 5-bromoindirubin-3'-methoxime, 5-bromoindirubin-3'-ethyloxime, 5,6-dichloroindirubin-3'-oximepropyl Various indirubin derivatives such as oxime and 6-chloroindirubin-3'-benzyloxime exist, but it was confirmed that the indirubin derivative represented by Formula 1 or Formula 2 was confirmed to have a significant effect on wound healing and recovery ( Experimental Example 1).

On the other hand, treatment of the indirubin derivative of Formula 1 or 2 through an in vitro (in virto ) experiment improves the wound closure rate (%) compared to the case of treatment with other conventional indirubin derivatives, and the wound healing effect is also simple. It has a healing effect (1.5 times, 2.0 times) more than a synergistic effect, and it was confirmed that the wound is being healed the fastest. Therefore, it was confirmed through an experiment that the indirubin derivative of Formula 1 or Formula 2 is the most preferable among conventional indirubin derivatives.

In addition, as a result of applying the indirubin derivative of Formula 1 or Formula 2 to the wound of the mouse, the acute wound in the mouse was re-epithelialized equal to or faster than the positive control EGF, and various kinds of collagen tissues were formed. It can be seen that the wound healing markers increase. Specifically, in the case of the indirubin derivative (A3334) of Formula 1, it exhibits a slightly better wound healing effect than the positive control EGF, but the epidermal layer of the skin is formed so that no traces such as dents or dents are left as the wound heals. It was confirmed that it is more effective in'scar improvement' that recovers evenly.

In addition, in the case of the indirubin derivative (A3051) of Formula 2, it was confirmed that it exhibits a wound recovery effect of 2 to 6 times or more at the same time as that of the positive control EGF, and through this, it can be seen that the wound recovers more quickly. In addition, it was confirmed that the epidermal layer of the skin recovered cleanly and uniformly so that no traces such as dents or dents were left as the wound healed than when treated with the positive control EGF. It was confirmed that it is also very effective in'improvement'.

In other words, it was confirmed that the indirubin derivative of Formula 1 or Formula 2, unlike conventional wound treatment agents or indirubin derivatives, has an unexpected and remarkable effect of not only healing wounds but also improving scars that induce uniform recovery of the epidermal layer of the skin.

The present invention may be a complex further comprising the extract of Shiva, and may be a complex further comprising any one or more low-molecular compounds selected from the group consisting of methyl vanylate, hesperidin, and quercitrin.

First, in the case of a composite obtained by mixing a Schizophrenia extract and an indirubin derivative at a certain ratio, it exhibits an excellent effect on wound healing. Specifically, when the indirubin derivative and the extract were treated together in an in-virto experiment, the wound closure rate (%) was improved compared to the case where the indirubin derivative of Formula 1 or Formula 2 was treated alone, It was confirmed that the wound healing effect also has a healing effect (2 times) more than a simple synergistic effect.

In particular, when treated with only the extract of Schizophrenia alone and when treated with the indirubin derivative alone, the values were about 1.5 to 2 times higher when the complex of Example 10 was treated than the sum of each. That is, it was confirmed that the composition for treating wounds of the present invention has a remarkable effect than that of using the extract and indirubin derivatives respectively, which exceeds the predictable value simply summed up the values obtained from each composition (Experimental Example 5 ).

In addition, it can be seen from the animal experiments in Experimental Example 6 that the complex with the indirubin derivative has a remarkable effect than the Shimane extract or the indirubin derivative alone, so it is more preferable.

The extract may be made of stems, leaves, fruits, pulverized products thereof, and mixtures thereof as a raw material, and the raw material is water, an anhydrous or hydrated lower alcohol having 1-4 carbon atoms, and a mixed solvent of the lower alcohol and water , Acetone, ethyl acetate, butyl acetate, chloroform, and may be extracted using a solvent selected from the group consisting of 1,3-butylene glycol as an extraction solvent.

The present invention may be a composite further comprising any one or more selected from the group consisting of methyl vanylate, hesperidin, and quercitrin. In the case of a composite obtained by mixing the low-molecular compound and the indirubin derivative at a certain ratio, it is excellent for wound healing. Shows the effect. Specifically, when the indirubin derivative and any one or more selected from the group consisting of methyl vanylate, hesperidin, and quercitrin were treated together in an in vitro (in virto) experiment, the indirubin derivative of Formula 1 or Formula 2 was used alone. Compared to the treated case, the wound closure rate (%) was improved, the wound healing effect also had a healing effect (1.4 to 2.3 times) more than a simple synergistic effect, and it was confirmed that the wound healed 6 hours faster. In other words, it was confirmed through an experiment that it is most preferable to use an indirubin derivative together with any one or more of any one or more selected from the group consisting of methyl vanylate, hesperidin, and quercitrin.

The composite in which any one or more selected from the group consisting of methyl vanylate, hesperidin, and quercitrin and any one or more indirubin derivatives selected from those represented by Formula 1 or Formula 2 are mixed, as in Experimental Examples to be described later. , Methyl vanylate, hesperidin, and quercitrin alone or a mixture of one or more indirubin derivatives selected from among those represented by the formula (2) as a result of analyzing the cell mobility, it was confirmed that the cell mobility was restored in 18 hours. . In particular, when the methyl vanylate and the indirubin derivative (A3051) were mixed, it was confirmed that the result was 5 times higher than that of the control group (cell) that was not treated with anything.

Any one or more low-molecular compounds selected from the group consisting of methyl vanylate, hesperidin, and quercitrin is a component contained in Shimane extract, Earth japonica extract, citrus fruit extract, chinensis extract, Yeoseongcho extract, and Eoseongcho extract, the extract , If any one or more stems, leaves, fruits, pulverized products thereof, and mixtures thereof selected from one or more stems, leaves, fruits, pulverized products thereof, and mixtures thereof selected from earth japonica, tangerine, skewer, and Eoseongcho are used as raw materials, and specifically, the raw material is water, carbon number 1- Anhydrous or water-containing lower alcohol of 4, a mixed solvent of the lower alcohol and water, acetone, ethyl acetate, butyl acetate, chloroform, and a solvent selected from the group consisting of 1,3-butylene glycol as an extraction solvent. I can.

The methyl vanylate is mainly present in the extract of Earthquake, and the hesperidin is present in the extract of Shimane tree and the extract of citrus, and the quercitrin may be mainly present in the extract of Yeokki, Pyeonchuk extract, and Eoseongcho extract.

When the composition further includes a serrata extract, the mixing weight ratio of the indirubin derivative represented by Formula 1 or 2 and the extract is not particularly limited thereto, but the extract and the indirubin derivative are based on 10 dry parts by weight of the extract. It is preferable that the indirubin derivative is mixed in an amount of 5 to 15 parts by weight, and if it is out of the above range, the increase in wound healing effect may be insignificant.

In addition, when the composition further comprises any one or more selected from the group consisting of methyl vanylate, hesperidine and quercitrin, the group consisting of the indirubin derivative represented by Formula 1 or 2 and the methyl vanylate, hesperidin and quercitrin The mixing weight ratio of any one or more low-molecular compounds selected from is not particularly limited thereto, but the low-molecular compound and the indirubin derivative are preferably mixed in a 1 to 20 molar ratio of the indirubin derivative based on 1 mol of the low-molecular compound, If it is out of the above range, the increase in wound healing effect may be insignificant.

The concentration in which the extract or the low-molecular compound is contained in the composition is not particularly limited as long as it is a general range in the art, but the extract or the low-molecular compound is 0.001 to 5% by weight or 0.001 in liquid weight based on the total weight of the composition. To 50% by weight, if less than 0.01% by weight, a sufficient wound healing effect may not appear, and if it exceeds 5% by weight (dry weight) or 50% by weight (liquid weight), skin irritation may be induced. have.

The food composition of the present invention may be prepared in the form of various foods, such as beverages, gums, teas, vitamin complexes, powders, granules, tablets, capsules, confectionery, rice cakes, and bread. Since the food composition of the present invention is composed of plant extracts with little toxicity and side effects, it can be safely used even when taken for a long time for prophylactic purposes.

When the indirubin derivative of the present invention is included in the food composition, the amount may be added in a proportion of 0.001 to 80% of the total weight.

Here, when the food composition is prepared in the form of a beverage, there is no particular limitation other than containing the food composition in the indicated ratio, and as an additional component, various flavoring agents or natural carbohydrates, etc. may be included as in ordinary beverages. That is, as natural carbohydrates, monosaccharides such as glucose, disaccharides such as fructose, and common sugars such as sucrose and polysaccharides, dextrins, cyclodextrins, and sugar alcohols such as xylitol, sorbitol, and erythritol can do. Examples of the flavoring agent include natural flavoring agents (taumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.).

In addition, the food composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavors and natural flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners. , pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonates used in carbonated beverages, and the like.

These components may be used independently or in combination. Although the proportion of these additives does not correspond to the essential element of the present invention, it is generally selected from 0.1 to about 50 parts by weight per 100 parts by weight of the food composition of the present invention.

The food composition may preferably be an emulsion formulation further comprising an oil, a surfactant, and polyethylene glycol.

In the emulsion formulation, the mixing weight ratio of the oil, surfactant, and polyethylene glycol may be 0.3-30:1:2-2.5, more preferably 10-20:1:2-2.5. The composition of the emulsion formulation can be determined by a pseudo three-phase diagram created according to a conventional method of determining the composition of the emulsion system. Specifically, after thoroughly mixing the oil phase (polyethoxylated castor oil (Kolliphor® EL)) and a surfactant (for example a mixture of tween 80 and polyethylene glycol) in different weight ratios in a specific ratio range, A pseudo three-phase diagram can be created by adding water to each proportion of the oil and surfactant mixture and marking the points corresponding to the emulsion-forming areas. By determining the emulsion region from the created pseudo-three-phase diagram and selecting a specific composition among the compositions contained in this region, the active ingredient (a complex of indirubin derivative or indirubin derivative and serrata extract, methyl vanylate, hesperidin and quercitrin) was prepared. It can be determined by the composition of the emulsion formulation to dissolve.

If the composition is in an emulsion formulation, the active ingredient (a complex of indirubin derivative or indirubin derivative and Japanese oak extract, methyl vanillate, hesperidin, and quercitrin) is preferably contained in an amount of 1 to 20% by weight based on the total weight of the composition. Do.

The surfactant is a Tween-based nonionic surfactant that acts as a co-solubilizer, and is used as an emulsifier or wetting agent for oral or parenteral preparations in pharmaceuticals, and is also used as an additive in cosmetics and foods. It is also used as a substance for suppressing p-glycoprotein in order to increase the bioavailability of drugs. Tween-based surfactants include polyoxyethylene sorbitan monolaurate (Tween 20), polyoxyethylene sorbitan monopalmitate (Tween 40), and polyoxyethylene sorbitan monolaurate (Tween 40). polyoxyethylene sorbitan monostearate, Tween 60) and polyoxyethylene sorbitan oleate (Tween 80). The tween-based surfactant is a stable substance approved for use by the US Food and Drug Administration for intravenous injection of the human body.

In addition, the polyethylene glycol is an amphoteric polymer having hydrophilicity and hydrophobicity, and the polyethylene glycol is a liquid when it has a low molecular weight, but becomes a solid as the molecular weight increases. The polyethylene glycol may be any one selected from the group consisting of PEG 150, 300, 400, 1000, 6000, 8000, 10000, 20000, 30000, and 40000. In this case, the PEG300 refers to polyethylene glycol having a molecular weight of 300. And, polyethylene glycol having a molecular weight exceeding 10,000 is also referred to as polyethylene oxide (PEO). Among these, PEG400 exists in liquid form and is frequently used for solubilization of various poorly soluble drugs, and is especially used for oral and parenteral (intravenous injection, subcutaneous injection, intramuscular injection, etc.) of the human body by the US Food and Drug Administration (FDA). It is a stable material that has been approved for.

The surfactant used in the examples of the present invention is polyoxyethylene sorbitan oleate (Tween 80), and polyethylene glycol is PEG 400, which has amphiphilic properties and is non-ionic that can be dispersible in water. , To provide an emulsion formulation optimized for poorly soluble active ingredients (any one or more active ingredients selected from the group consisting of the indirubin derivative represented by Chemical Formula 1 or 2, Schizophrenia extract, methyl vanylate, hesperidin and quercitrin) I can.

The oil may be any one or more selected from the group consisting of polyethoxylated castor oil (Kolliphor® EL), sunflower oil, and olive oil.

The composition may contain cyclodextrin for higher solubilization. Cyclodextrin may be 100 to 1000 parts by weight based on 100 parts by weight of the active ingredient present in the composition.

The emulsion formulation is a stable formulation that does not show changes in wnt activity and solubility in distilled water under conditions of 25 to 4°C when measured at 3 months. It enables the activity of the active ingredient to be maintained for a long period of time and effectively absorbs into the body. It was confirmed that it can be improved.

In addition, for treating or improving wounds comprising any one or more selected from the group consisting of the indirubin derivative represented by the following Formula 1, the indirubin derivative represented by the Formula 2, the serrata extract, methyl vanylate, hesperidin, and quercitrin The manufacturing method of the emulsion food composition is as follows.

1) preparing a first solution by mixing oil, surfactant and polyethylene glycol;

2) a second solution containing at least one active ingredient selected from the group consisting of the indirubin derivative represented by the formula 1, the indirubin derivative represented by the formula 2, the chinensis extract, methyl vanylate, hesperidin, and quercitrin Manufacturing steps;

3) preparing an emulsion composition by mixing a second solution with the first solution.

The second solution may contain cyclodextrin for higher solubilization.

The polyethylene glycol is an amphoteric polymer having hydrophilicity and hydrophobicity, and the polyethylene glycol is a liquid when it has a low molecular weight, but becomes a solid as the molecular weight increases. The polyethylene glycol may be any one selected from the group consisting of PEG 150, 300, 400, 1000, 6000, 8000, 10000, 20000, 30000, and 40000. In this case, the PEG300 refers to polyethylene glycol having a molecular weight of 300. And, polyethylene glycol having a molecular weight exceeding 10,000 is also referred to as polyethylene oxide (PEO). Among these, PEG400 exists in liquid form and is frequently used for solubilization of various poorly soluble drugs, and is especially used for oral and parenteral (intravenous injection, subcutaneous injection, intramuscular injection, etc.) of the human body by the US Food and Drug Administration (FDA). It is a stable material that has been approved for.

The surfactant is a Tween-based nonionic surfactant that acts as a co-solubilizer, and is used as an emulsifier or wetting agent for oral or parenteral preparations in pharmaceuticals, and is also used as an additive in cosmetics and foods. It is also used as a substance for suppressing p-glycoprotein in order to increase the bioavailability of drugs. Tween-based surfactants include polyoxyethylene sorbitan monolaurate (Tween 20), polyoxyethylene sorbitan monopalmitate (Tween 40), and polyoxyethylene sorbitan monolaurate (Tween 40). polyoxyethylene sorbitan monostearate, Tween 60) and polyoxyethylene sorbitan oleate (Tween 80). The tween-based surfactant is a stable substance approved for use by the US Food and Drug Administration for intravenous injection of the human body.

The mixing weight ratio of the oil, surfactant and polyethylene glycol may be 0.3-30:1:2-2.5, more preferably 10-20:1:2-2.5. If the composition is in an emulsion formulation, the active ingredient (a complex of indirubin derivative or indirubin derivative and serrata extract, methyl vanillate, hesperidin, and quercitrin) is contained in an amount of 1 to 20% by weight based on the total weight of the composition. desirable.

When water was added to the emulsion composition of the present invention to take a microscopic picture and compared, the emulsion of the present invention was completely dissolved to form an emulsion formulation (F8) in a solution state. In addition, as a result of observing the stability of the emulsion of the present invention, the droplets have a spherical shape, the average diameter is 20 to 1500 nm, the preferred diameter is measured to be 30 to 50 nm to form nano-sized droplets, and a narrow range of size distribution Is shown.

In addition, it was confirmed that the emulsion of the present invention maintains stability for a long period of time because there is no change in solubility and activity of active ingredients for 3 months at room temperature. Furthermore, when the emulsion formulation of the present invention is administered orally or applied to a topical area, absorption can be effectively improved.

As can be seen in the following examples, the indirubin derivative represented by the following formula (1) or formula (2) exhibits not only skin regeneration, but also excellent collagen synthesis promoting effect, and therefore, pharmaceuticals, cosmetics, and health care for skin regeneration and elasticity enhancement. It can be used as an active ingredient such as food.

Accordingly, another aspect of the present invention relates to a composition for skin regeneration and elasticity enhancement comprising an indirubin derivative represented by the following Formula 1 or Formula 2 as an active ingredient.

[Formula 1]

[Formula 2]

The indirubin derivative of the present invention is a substance that shows little toxicity by cells even if it is treated for a long period of time, and has excellent cell regeneration, wound healing, and collagen production effects, thereby preventing or treating wounds, as well as skin regeneration and It can be seen that it can be applied to various fields such as pharmacy, food, and cosmetics with elasticity enhancing function.

In this specification, "skin regeneration" means reducing, removing, alleviating, and improving damage to the skin, and "improving elasticity" means maintaining and strengthening the ability related to elasticity of the skin. . Collagen, a collagen fiber in the dermal layer of the skin, is a major protein that plays a role in skin elasticity, and the biosynthesis of collagen is affected internally and externally. Specifically, in skin cells, when the cellular activity decreases due to natural aging, collagen fibers decrease, or as an external factor, excessive irradiation of ultraviolet rays or damage to the skin, or active oxygen generated by stress, etc. By reacting with thiol: -SH), enzyme activity is inhibited, or by increasing the expression of degrading enzymes such as collagen, skin wrinkles are increased and elasticity is reduced, leading to skin aging.

The indirubin derivatives include 5-methoxyl indirubin-3'-oxime, 5-methoxyl indirubin-3'-methoxime, indirubin-3'-oxime, 6-bromoindirubin-3'-oxime , 5,6-dichloroindirubin, 5,6-dichloroindirubin-3'-oxime, 5,6-dichloroindirubin-3'-methoxime, 5,6-dichloroindirubin-3'-propyloxime, 6-chloro-5-nitroindirubin, 6-chloro-5-nitroindirubin-3'-oxime, 6-chloroindirubin-3'-methoxime, 5-chloroindirubin-3'-methoxime, 5 -Bromoindirubin-3'-oxime, 5-bromoindirubin-3'-methoxime, 5-bromoindirubin-3'-ethyloxime, 5,6-dichloroindirubin-3'-oximepropyl Various indirubin derivatives such as oxime and 6-chloroindirubin-3'-benzyloxime exist, but it was confirmed that the indirubin derivative represented by Formula 1 or Formula 2 was confirmed to have a significant effect on cell recovery (Experimental Example One).

As a result of applying the indirubin derivative of Formula 1 or Formula 2 to the wound of the mouse, it was found that the acute wound in the mouse was re-epithelialized equal to or faster than the positive control EGF, and various kinds of collagen activity were enhanced. I can confirm. Therefore, it was confirmed that the composition according to the present invention has an effect of not only maximizing cell motility, but also promoting and accumulating collagen in keratinocytes and fibroblasts that play an important role in wound healing. It can be seen that it can be used as an active ingredient in medicines, cosmetics, and health foods for promotion.

The indirubin derivative used in the present invention can be provided not only as a free substance, but also as a pharmaceutically acceptable salt, pharmaceutically acceptable solvate, pharmaceutically acceptable polymorph, or pharmaceutically acceptable prodrug thereof. have. The salt of the indirubin derivative is not particularly limited as long as it can be blended in medicine or cosmetics. It includes an inorganic salt or an organic salt, and may be an acid salt or an alkaline salt. In particular, in the case of a cation salt, alkali metal salts such as sodium salts and potassium salts, alkaline earth metal salts such as calcium salts, magnesium salts and barium salts; Basic amino acid salts such as arginine and lysine; Ammonium salts such as ammonium salts and tricyclohexyl ammonium salts; And various alkanol amine salts such as monoethanolamine salt, diethanolamine salt, triethanolamine salt, monoisopropanolamine salt, diisopropanolamine salt, and triisopropanolamine. Preferably, the salt is an alkali metal salt, more preferably a tetrasodium salt.

In the composition, the indirubin derivative may be included in an amount of 0.001 to 80% by weight based on the total weight of the composition, and if it is less than 0.01% by weight, sufficient wound prevention or healing effect may not appear, and when it exceeds 80% by weight, it is excessively included. May cause skin irritation.

The present invention may be a complex further comprising the extract of Shiva, and may be a complex further comprising any one or more low-molecular compounds selected from the group consisting of methyl vanylate, hesperidin, and quercitrin.

First, in the case of a composite obtained by mixing a Schizophrenia extract and an indirubin derivative at a certain ratio, it exhibits an excellent effect on wound healing. Specifically, when the indirubin derivative and the extract were treated together in an in-virto experiment, the wound closure rate (%) was improved compared to the case where the indirubin derivative of Formula 1 or Formula 2 was treated alone, It was confirmed that the wound healing effect also has a healing effect (2 times) more than a simple synergistic effect.

In particular, when treated with only the extract of Schizophrenia alone and when treated with the indirubin derivative alone, the values were about 1.5 to 2 times higher when the complex of Example 10 was treated than the sum of each. That is, it was confirmed that the composition for treating wounds of the present invention has a remarkable effect than that of using the extract and indirubin derivative respectively, which exceeds the predictable value obtained by simply summing the values obtained from each composition (Experimental Example 5 ).

In addition, it can be seen from the animal experiments in Experimental Example 6 that the complex with the indirubin derivative has a remarkable effect than the Shimane extract or the indirubin derivative alone, so it is more preferable.

The extract may be made of stems, leaves, fruits, pulverized products thereof, and mixtures thereof as a raw material, and the raw material is water, an anhydrous or hydrated lower alcohol having 1-4 carbon atoms, and a mixed solvent of the lower alcohol and water , Acetone, ethyl acetate, butyl acetate, chloroform, and may be extracted using a solvent selected from the group consisting of 1,3-butylene glycol as an extraction solvent.

When the composition further includes a serrata extract, the mixing weight ratio of the indirubin derivative represented by Formula 1 or 2 and the extract is not particularly limited thereto, but the extract and the indirubin derivative are based on 10 dry parts by weight of the extract. It is preferable that the indirubin derivative is mixed in an amount of 5 to 15 parts by weight, and if it is out of the above range, the increase in wound healing effect may be insignificant.

The present invention may be a composite further comprising any one or more selected from the group consisting of methyl vanylate, hesperidin and quercitrin, and a composite obtained by mixing the methyl vanylate, hesperidine and quercitrin and indirubin derivatives at a certain ratio , It shows excellent effect on skin regeneration and elasticity enhancement. Specifically, when the indirubin derivative and methyl vanylate, hesperidine, and quercitrin were treated together in an in-virto experiment, compared to the case of treatment with the indirubin derivative of Formula 1 or Formula 2 alone, the wound closure rate It was confirmed that the (%) improved, the wound healing effect also had a skin regeneration effect (1.4 to 2.3 times) more than a simple synergistic effect, and improved the motility of carotenocytes 6 hours faster. In other words, it was confirmed through an experiment that it is most preferable to use an indirubin derivative together with any one or more of the methyl vanylate, hesperidin and quercitrin.

The compound in which the methyl vanylate, hesperidin, and quercitrin and any one or more indirubin derivatives selected from those represented by Formula 1 or Formula 2 are mixed, as in the experimental examples described later, methyl vanylate, hesperidine, and quercitrin As a result of analyzing the cell mobility by the complex mixture containing one or more indirubin derivatives selected from alone or in science and formula 2, it was confirmed that the cell mobility was restored in 18 hours. In particular, when the methyl vanylate and the indirubin derivative (A3051) were mixed, it was confirmed that the result was 5 times higher than that of the control group (cell) that was not treated with anything.

Any one or more low-molecular compounds selected from the group consisting of methyl vanylate, hesperidin, and quercitrin is a component contained in Shimane extract, Earth japonica extract, citrus fruit extract, chinensis extract, Yeoseongcho extract, and Eoseongcho extract, the extract , If any one or more stems, leaves, fruits, pulverized products thereof, and mixtures thereof selected from one or more stems, leaves, fruits, pulverized products thereof, and mixtures thereof selected from earth japonica, tangerine, skewer, and Eoseongcho are used as raw materials, and specifically, the raw material is water, carbon number 1- Anhydrous or water-containing lower alcohol of 4, a mixed solvent of the lower alcohol and water, acetone, ethyl acetate, butyl acetate, chloroform, and a solvent selected from the group consisting of 1,3-butylene glycol as an extraction solvent. I can.

The methyl vanylate is mainly present in the extract of Earthquake, and the hesperidin is present in the extract of Shimane tree and the extract of citrus, and the quercitrin may be mainly present in the extract of Yeokki, Pyeonchuk extract, and Eoseongcho extract.

When the composition further comprises any one or more selected from the group consisting of methyl vanylate, hesperidine and quercitrin, selected from the group consisting of the indirubin derivative represented by Formula 1 or 2 and the methyl vanylate, hesperidine and quercitrin The mixing weight ratio of any one or more of the low molecular weight compounds is not particularly limited thereto, but the low molecular weight compound and the indirubin derivative are preferably mixed in a 1 to 20 mole ratio of the indirubin derivative based on 1 mole of the low molecular compound, and the If it is out of range, the increase in wound healing effect may be insignificant.

The concentration in which the extract or the low-molecular compound is contained in the composition is not particularly limited as long as it is a general range in the art, but the extract or the low-molecular compound is 0.001 to 5% by weight or 0.001 in liquid weight based on the total weight of the composition. To 50% by weight, if less than 0.01% by weight, a sufficient wound healing effect may not appear, and if it exceeds 5% by weight (dry weight) or 50% by weight (liquid weight), skin irritation may be induced. have.

In the cosmetic composition according to the present invention, an additive capable of increasing the cosmetic effect through wound improvement may be further included, but is not particularly limited thereto.

Cosmetic composition comprising the indirubin derivative according to the present invention as an active ingredient is a lotion, essence. It can be prepared in lotion, cream, pack, gel, powder, foundation or detergent formulation, for example, lotion, nutrition lotion, nutrition essence, massage cream, beauty bath water additive, body lotion, body milk, bath oil, baby oil, Baby powder, shower gel, shower cream, sunscreen lotion, sunscreen cream, suntan cream, skin lotion, skin cream, sunscreen cosmetics, cleansing milk, depilatory makeup, face and body lotion, face and body cream, skin whitening Cream, hand lotion, hair lotion, cosmetic cream, jasmine oil, bath soap, water soap, beauty soap, shampoo, hand sanitizer (hand cleaner), medicinal soap, non-medical, cream soap, facial wash, body cleaner, scalp cleaner, hair It can be prepared in the form of rinse, cosmetic soap, tooth whitening gel, toothpaste, and the like. To this end, the composition of the present invention may further include a solvent or a suitable carrier, excipient, or diluent that is commonly used in the manufacture of a cosmetic composition.

The type of solvent that can be further added to the cosmetic composition of the present invention is not particularly limited, but, for example, water, saline, DMSO, or a combination thereof may be used, and as a carrier, excipient or diluent, purified water, oil, wax , Fatty acids, fatty alcohols, fatty acid esters, surfactants, humectants, thickeners, antioxidants, viscosity stabilizers, chelating agents, buffers, lower alcohols, and the like, but are not limited thereto. In addition, whitening agents, moisturizing agents, vitamins, sunscreen agents, perfumes, dyes, antibiotics, antibacterial agents, and antifungal agents may be included as needed.

Hydrogenated vegetable oil, castor oil, cottonseed oil, olive oil, palm seed oil, jojoba oil, and avocado oil may be used as the oil, and waxes include beeswax, spermaceti, carnauba, candelilla, montan, ceresin, liquid paraffin, and lanolin. Can be.

Stearic acid, linoleic acid, linolenic acid, oleic acid may be used as the fatty acid, and cetyl alcohol, octyldodecanol, oleyl alcohol, panthenol, lanolin alcohol, stearyl alcohol, hexadecanol may be used as fatty alcohols. In addition, as the fatty acid ester, isopropyl myristate, isopropyl palmitate, and butyl stearate may be used. In addition, as the surfactant, cationic surfactants, anionic surfactants, and nonionic surfactants known in the art can be used, and surfactants derived from natural products are preferred as far as possible.

In addition, hygroscopic agents, thickeners, antioxidants, and the like, which are widely known in the cosmetic field may be included, and the types and amounts thereof are as known in the art.

However, the composition of the present invention may be used by a method such as direct application or dispersion to the skin.

The present invention also relates to a food composition for skin regeneration and elasticity improvement, comprising the indirubin derivative represented by Formula 1 or Formula 2 as an active ingredient.

The food composition may be formulated in the form of a capsule, tablet, powder, granule, liquid, pill, flake, paste, syrup, gel, jelly, or bar of a composition containing the indirubin derivative, or It is added to food materials such as beverages, teas, spices, chewing gum, and confectionery, and is manufactured in the form of a general food. When ingested, it means that it has a specific effect on health, but unlike general drugs, it is made from food as a raw material. It has the advantage of not having side effects that may occur when taking the drug for a long time.

The food composition is very useful because it can be consumed on a daily basis. The addition amount of the indirubin derivative in such a food composition varies depending on the type of the target food composition and cannot be uniformly defined, but it may be added within a range that does not impair the original taste of the food, and is usually 0.001 with respect to the target food. To 50% by weight, preferably 0.1 to 20% by weight. In addition, in the case of health functional foods in the form of capsules, tablets, powders, granules, liquids, pills, flakes, pastes, syrups, gels, jelly or bars, usually 0.1 to 100% by weight, preferably 0.5 to 80% by weight. It can be added in the range of %.

The food composition may include not only the indirubin derivative as an active ingredient, but also ingredients that are commonly added during food production, and include, for example, proteins, carbohydrates, fats, nutrients, flavoring agents, and flavoring agents. Examples of the aforementioned carbohydrates include monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides, and the like; And polysaccharides, for example, common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol.

As flavoring agents, natural flavoring agents [taumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.]) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the health functional food of the present invention is made of drinks and beverages, in addition to the composition of the present invention, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, and various plant extracts may be additionally included.

The skin regeneration or elasticity enhancing composition of the present invention may preferably be an emulsion formulation further comprising an oil, a surfactant, and polyethylene glycol.

In the emulsion formulation, the mixing weight ratio of the oil, surfactant, and polyethylene glycol may be 0.3-30:1:2-2.5, more preferably 10-20:1:2-2.5. The composition of the emulsion formulation can be determined by a pseudo three-phase diagram created according to a conventional method of determining the composition of the emulsion system. Specifically, after thoroughly mixing the oil phase (polyethoxylated castor oil (Kolliphor® EL)) and a surfactant (for example a mixture of tween 80 and polyethylene glycol) in different weight ratios in a specific ratio range, A pseudo three-phase diagram can be created by adding water to each proportion of the oil and surfactant mixture and marking the points corresponding to the emulsion-forming areas. Determine the emulsion region from the created pseudo-three-phase diagram, select a specific composition among the compositions contained in this region, and use the above active ingredients (complex of indirubin derivative or indirubin derivative and Schizophrenia extract, methyl vanylate, hesperidin and quercitrin). It can be determined by the composition of the emulsion formulation to dissolve.

If the composition is in an emulsion formulation, the active ingredient (a complex of indirubin derivative or indirubin derivative and Japanese oak extract, methyl vanillate, hesperidin, and quercitrin) is preferably contained in an amount of 1 to 20% by weight based on the total weight of the composition. Do.

The surfactant is a Tween-based nonionic surfactant that acts as a co-solubilizer, and is used as an emulsifier or wetting agent for oral or parenteral preparations in pharmaceuticals, and is also used as an additive in cosmetics and foods. It is also used as a substance for suppressing p-glycoprotein in order to increase the bioavailability of drugs. Tween-based surfactants include polyoxyethylene sorbitan monolaurate (Tween 20), polyoxyethylene sorbitan monopalmitate (Tween 40), and polyoxyethylene sorbitan monolaurate (Tween 40). polyoxyethylene sorbitan monostearate, Tween 60) and polyoxyethylene sorbitan oleate (Tween 80). The tween-based surfactant is a stable substance approved for use by the US Food and Drug Administration for intravenous injection of the human body.

In addition, the polyethylene glycol is an amphoteric polymer having hydrophilicity and hydrophobicity, and the polyethylene glycol is a liquid when it has a low molecular weight, but becomes a solid as the molecular weight increases. The polyethylene glycol may be any one selected from the group consisting of PEG 150, 300, 400, 1000, 6000, 8000, 10000, 20000, 30000, and 40000. In this case, the PEG300 refers to polyethylene glycol having a molecular weight of 300. And, polyethylene glycol having a molecular weight exceeding 10,000 is also referred to as polyethylene oxide (PEO). Among these, PEG400 exists in liquid form and is frequently used for solubilization of various poorly soluble drugs, and is especially used for oral and parenteral (intravenous injection, subcutaneous injection, intramuscular injection, etc.) of the human body by the US Food and Drug Administration (FDA). It is a stable material that has been approved for.

The surfactant used in the examples of the present invention is polyoxyethylene sorbitan oleate (Tween 80), and polyethylene glycol is PEG 400, which has amphiphilic properties and is non-ionic that can be dispersible in water. , To provide an emulsion formulation optimized for poorly soluble active ingredients (any one or more active ingredients selected from the group consisting of the indirubin derivative represented by Chemical Formula 1 or 2, Schizophrenia extract, methyl vanylate, hesperidin and quercitrin) I can.

The oil may be any one or more selected from the group consisting of polyethoxylated castor oil (Kolliphor® EL), sunflower oil, and olive oil.

The composition may contain cyclodextrin for higher solubilization. Cyclodextrin may be 100 to 1000 parts by weight based on 100 parts by weight of the active ingredient present in the composition.

The emulsion formulation is a stable formulation that does not show changes in wnt activity and solubility in distilled water under conditions of 25 to 4°C when measured at 3 months. It enables the activity of the active ingredient to be maintained for a long period of time and effectively absorbs into the body. It was confirmed that it can be improved.

In addition, skin regeneration or elasticity enhancement comprising any one or more selected from the group consisting of the indirubin derivative represented by the following formula (1), the indirubin derivative represented by the formula (2), serrata extract, methyl vanylate, hesperidin, and quercitrin The preparation method of the emulsion composition is as follows.

1) preparing a first solution by mixing oil, surfactant and polyethylene glycol;

2) a second solution containing at least one active ingredient selected from the group consisting of the indirubin derivative represented by the formula 1, the indirubin derivative represented by the formula 2, the chinensis extract, methyl vanylate, hesperidin, and quercitrin Manufacturing steps;

3) preparing an emulsion composition by mixing a second solution with the first solution.

The second solution may contain cyclodextrin for higher solubilization.

The polyethylene glycol is an amphoteric polymer having hydrophilicity and hydrophobicity, and the polyethylene glycol is a liquid when it has a low molecular weight, but becomes a solid as the molecular weight increases. The polyethylene glycol may be any one selected from the group consisting of PEG 150, 300, 400, 1000, 6000, 8000, 10000, 20000, 30000, and 40000. In this case, the PEG300 refers to polyethylene glycol having a molecular weight of 300. And, polyethylene glycol having a molecular weight exceeding 10,000 is also referred to as polyethylene oxide (PEO). Among these, PEG400 exists in liquid form and is frequently used for solubilization of various poorly soluble drugs, and is especially used for oral and parenteral (intravenous injection, subcutaneous injection, intramuscular injection, etc.) of the human body by the US Food and Drug Administration (FDA). It is a stable material that has been approved for.

The surfactant is a Tween-based nonionic surfactant that acts as a co-solubilizer, and is used as an emulsifier or wetting agent for oral or parenteral preparations in pharmaceuticals, and is also used as an additive in cosmetics and foods. It is also used as a substance for suppressing p-glycoprotein in order to increase the bioavailability of drugs. Tween-based surfactants include polyoxyethylene sorbitan monolaurate (Tween 20), polyoxyethylene sorbitan monopalmitate (Tween 40), and polyoxyethylene sorbitan monolaurate (Tween 40). polyoxyethylene sorbitan monostearate, Tween 60) and polyoxyethylene sorbitan oleate (Tween 80). The tween-based surfactant is a stable substance approved for use by the US Food and Drug Administration for intravenous injection of the human body.

The mixing weight ratio of the oil, surfactant and polyethylene glycol may be 0.3-30:1:2-2.5, more preferably 10-20:1:2-2.5. If the composition is in an emulsion formulation, the active ingredient (a complex of indirubin derivative or indirubin derivative and serrata extract, methyl vanillate, hesperidin, and quercitrin) is contained in an amount of 1 to 20% by weight based on the total weight of the composition. desirable.

When water was added to the emulsion composition of the present invention to take a microscopic picture and compared, the emulsion of the present invention was completely dissolved to form an emulsion formulation (F8) in a solution state. In addition, as a result of observing the stability of the emulsion of the present invention, the droplets have a spherical shape, the average diameter is 20 to 1500 nm, the preferred diameter is measured to be 30 to 50 nm to form nano-sized droplets, and a narrow range of size distribution Is shown.

In addition, it was confirmed that the emulsion of the present invention maintains stability for a long period of time because there is no change in solubility and activity of active ingredients for 3 months at room temperature. Furthermore, when the emulsion formulation of the present invention is administered orally or applied to a topical area, absorption can be effectively improved.

Hereinafter, the present invention will be described in more detail through examples and the like, but the scope and contents of the present invention are reduced or limited by the following examples, and thus cannot be interpreted. In addition, if based on the disclosure of the present invention including the following examples, it is obvious that the present invention for which no specific experimental results are presented can be easily carried out by a person skilled in the art. It is natural to fall within the scope of the claims.

In addition, the experimental results presented below are only representative of the experimental results of the above examples and comparative examples, and the effects of each of the various embodiments of the present invention that are not explicitly presented below will be described in detail in the corresponding section.

Example 1. Synthesis of 5-methoxyl indirubin-3'-oxime (A3334)

① Intermediate 5'-methoxy-[2,3'-biindolinylidene]-2',3-dione (5'-methoxy-[2,3'-biindolinylidene]-2',3-dione) Synthesis of

5-methoxyisatin (1000 mg, 5.65 mmol) was added to a 250 ml round bottom flask, dissolved in methanol (MeOH) (225 ml), and then indoxyl acetate (989 mg, 5.65). mmol) and sodium carbonate (Na ₂ CO ₃ ) (1496 mg, 14.11 mmol) were added, followed by stirring at 65° C. for 12 hours. TLC (Rf=0.4, ethyl acetate/hexane = 1/2 (v/v)) was used to confirm the completion of the reaction, and the product was cooled on ice until a crystalline mass was formed. When crystals are formed, the solvent is removed by filtration, and the filtrate is discarded and the product is washed several times with a solvent (ethanol/water = 1/1(v/v)). The product was filtered, dried on a vacuum pump and used in the next step without further purification.

Synthesis of A3334

In a 100 ml round bottom flask, 5'-methoxy-[2,3'-biindolinylidene]-2',3-dione (5'-methoxy-[2,3'-biindolinylidene]-2', 3-dione) (670 mg, 2.29 mmol) was added, dissolved in pyridine (27 mL), and H ₂ NOHHCl (3186 mg, 45.85 mmol) was added, followed by stirring at 120° C. for 12 hours. do. TLC (R _f =0.5, ethyl acetate/hexane = 1/1 (v/v)) was used to confirm the completion of the reaction and lower the temperature of the reaction solution to room temperature. After evaporating all of the pyridine solvent of the product, add water and ethyl acetate to dissolve the product using ultrasonic waves for 30 minutes, then extract twice with ethyl acetate and wash with a saturated solution of _{sodium chloride (NaHCO 3 ).} After the extracted solution is dehydrated with anhydrous magnesium sulfate, the solvent is evaporated and recrystallized using methanol and nucleic acid. When the product is dried in a vacuum pump, a red solid A3334 (420 mg) can be obtained in a yield of 59%. ¹ H NMR (400 MHz, DMSO- d6 ) δ 13.52 (s, 1H), 11.79 (s, 1H), 10.54 (s, 1H), 8.35 (d, J = 1.5 Hz, 1H), 8.26 (d, J = 7.6 Hz, 1H), 7.41 (d, J = 2.8 Hz, 2H), 7.08-7.00 (m, 1H), 6.82-6.71 (m, 2H), 3.78 (s, 3H).

Example 2. Synthesis of 5,6-dichloroindirubin-3'-methoxime (A3051).

① Intermediate 5',6'-dichloro-[2,3'-biindolinylidene]-2'3-dione (5',6'-dichloro-[2,3'-biindolinylidene]-2' ,3-dione) synthesis

5,6-dichloroisatin (500 mg, 2.32 mmol) was added to a 250 ml round-bottom flask, dissolved in methanol (MeOH) (92.80 ml), and then indoxyl acetate (indoxyl acetate). 405.48 mg, 2.315 mmol) sodium percarbonate (Na ₂ CO ₃ ) (637.83 mg, 6.02 mmol) was added, followed by stirring at 65° C. for 12 hours. TLC (Rf=0.4, ethyl acetate/hexane = 1/2 (v/v)) was used to confirm the completion of the reaction, and the product was cooled on ice until a crystalline mass was formed. When crystals are formed, the solvent is removed by filtration, and the filtrate is discarded and the product is washed several times with a solvent (ethanol/water = 1/1(v/v)). The product was filtered, dried on a vacuum pump and used in the next step without further purification.

② Synthesis of A3051

In a 100 ml round bottom flask, 5',6'-dichloro-[2,3'-biindolinylidene]-2'3-dione (5',6'-dichloro-[2,3'-biindolinylidene) ]-2',3-dione) (600 mg, 1.81 mmol) was added, dissolved in pyridine (151 mL), and H ₂ NOCH ₃ ㅇHCl (3026.4 mg, 36.24 mmol) was added, followed by stirring at 120° C. for 12 hours. TLC (R _f =0.4, ethyl acetate/hexane = 1/1 (v/v)) was used to confirm the completion of the reaction and lower the temperature of the reaction solution to room temperature. After evaporating all of the pyridine solvent of the product, add water and ethyl acetate to dissolve the product using ultrasonic waves for 30 minutes, then extract twice with ethyl acetate and wash with a saturated solution of _{sodium chloride (NaHCO 3 ).} After the extracted solution is dehydrated with anhydrous magnesium sulfate, the solvent is evaporated and recrystallized using methanol and nucleic acid. When the product is dried with a vacuum pump, a red solid A3051 (326 mg) can be obtained in a yield of 47.94%. ¹ H NMR (400 MHz, DMSO- d6 ) δ 11.36 (s, 2H), 8.80 (s, 1H), 8.08 (d, 1H, J = 7.7 Hz), 7.46-7.41 (m, 2H), 7.07-6.99 (m, 2H), 4.38 (s, 3H).

Example 3. Preparation of methyl vanillate

Methyl vanillate was purchased and used from sigma aldrich.

Example 4. Hesperidin preparation

Hesperidin was purchased and used from sigma aldrich.

Example 5. Quercitrin preparation

Quercitrin was purchased and used from sigma aldrich.

The powder of the Eoseongcho extract was dissolved in dimethyl sulfide (DMSO) at a concentration of 1 μg/ml and used in the experiment.

Example 6. Preparation of Schizophrenia extract

Shrub extract was purchased and used from Korea Plant Extract Bank (KOREA PLANT EXTACT BANK).

Examples 7 to 9. Composite preparation.

A composite was prepared by mixing the methyl vanylate prepared from Example 3, the hesperidin of Example 4, the quercitrin of Example 5, and the indirubin derivative (A3051) represented by Chemical Formula 2 of Example 2 in an appropriate ratio.

Example 10. Preparation of a composite material of the extract of Schizandra chinensis and indirubin derivative (A3486)

A composite of 0.1 µM, 1 µM, and 0.5 mM of the indirubin derivative (A3486) represented by the formula 1 of Example 1 with 0.1 µM, 1 µM, and 0.5 mM of the extract of Schizandra chinensis prepared in Example 6 Was prepared. The mixing ratio of the extract and the indirubin derivative was described in each of the experimental examples.

Example 11. Preparation of a composite material of the extract and indirubin derivatives

A composite of 0.1 µM, 1 µM, and 0.5 mM of the indirubin derivative (A3051) represented by Chemical Formula 2 of Example 2 with 0.1 µM, 1 µM, and 0.5 mM of the extract of Schizandra chinensis prepared in Example 6 Was prepared. The mixing ratio of the extract and the indirubin derivative was described in each of the experimental examples.

Comparative Example 1. Indirubin-3'-oxime (I3O or IO)

[Formula 3]

Comparative Example 2. 5,6-dichloroindirubin (A2941)

[Formula 4]

5,6-dichloroindirubin of Formula 6 is an intermediate of the indirubin derivative of Example 2 (Chemical Formula 2), and was prepared in the same manner as in Example 2-①.

Comparative Example 3. 5,6-dichloroindirubin-3'-oxime (A3050)

[Formula 5]

Comparative Example 4. 6-chloro-5-nitroindirubin-3'-oxime (A2736)

[Formula 6]

6-Chloro-5-nitroindirubin (A2735) (350mg, 1.02mmol) was added to a 50ml round bottom flask, dissolved in pyridine (7 ml), and H2NOHHCl (711.78 mg, 10.24 mmol) was added, followed by 120°C. Reflux for about 12 hours above. TLC (Rf=0.5, ethyl acetate/hexane= 1/1 (v/v)) was used to confirm the completion of the reaction, and the temperature of the reaction solution was lowered to room temperature. After evaporating all of the pyridine solvent of the product, add water and ethyl acetate to dissolve the product using ultrasonic waves for 30 minutes, then extract twice with ethyl acetate and wash with a saturated solution of sodium chloride (NaHCO3). After the extracted solution was dehydrated with anhydrous magnesium sulfate, the solvent was evaporated and recrystallized using methanol and nucleic acid. When the product is dried in a vacuum pump, a red solid A2736 (284 mg) can be obtained in a yield of 78%. 1H NMR (400 MHz, DMSO-d6) δ 13.81 (s, 1H), 11.77 (s, 1H), 11.30 (s, 1H), 9.13 (s, 1H), 8.16 (d, J = 8.0 Hz, 1H) , 7.42-7.35 (m, 2H), 7.04 (t, J = 8.0 Hz, 1H), 6.99 (s, 1H).

Comparative Example 5. 6-Chloroindirubin-3'-methoxime (A2793)

[Formula 7]

Comparative Example 6. 5-Bromoindirubin-3'-methoxime (A3391)

[Formula 8]

Comparative Example 7. 5-Methoxyl indirubin-3'-methoxime (A3441)

[Formula 9]

Experimental Example 1. Confirmation of cell motility.

To compare the mobility of cells by the indirubin derivatives prepared from Examples 1 and 2 and the indirubin derivatives prepared from Comparative Examples 1 to 7.

In wound healing, HaCaT cells, that is, human keratinocyte cells, which play an important role in wound healing, were cultured in a 12-well plate for 24 hours. In this case, DMEM containing 10% FBS was used. After 24 hours of incubation, the medium was exchanged with DMEM medium containing 5% FBS, and then a scratch was made in the center of the cells using a 1000p (blue) tip. Thereafter, the indirubin derivatives of Examples 1 and 2 and Comparative Examples 1 to 7 were treated at a concentration of 5 μg/ml and 1 μM, respectively, on the scratched area, and cell motility was observed after 24 hours.

1 is a photograph showing the mobility of cells in keratinocytes in order to confirm the mobility of cells according to the treatment of the indirubin derivatives prepared from Examples 1, 2, and Comparative Examples 1 to 7, and FIG. 2 Is a graph quantitatively showing the cell migration rate in keratinocyte cells in order to confirm the mobility of cells according to the treatment of the indirubin derivatives prepared from Examples 1, 2, and Comparative Examples 1 to 7. At this time, the control refers to keratinocyte cells not treated with the indirubin derivative.

In FIG. 2, the wound closure rate (%) represents the results (FIG. 1) of each treatment of the indirubin derivatives prepared in Examples 1 and 2 and Comparative Examples 1 to 7 to keratinocytes, which play an important role in wound healing. , It is expressed by quantification by cell migration rate.

As shown in FIG. 1, it was confirmed that the indirubin derivatives prepared from Comparative Examples 1 to 7 had no difference in cell motility compared to the control cells.

On the other hand, it can be seen that the motility in the cells treated with the indirubin derivatives prepared from Examples 1 and 2 was improved by up to 1.5 to 2 times or more than the motility in the control cells. In addition, when each of the indirubin derivatives A3334 and A3051 of Examples 1 and 2 were treated, it was confirmed that the motility of the cells of keratinocytes, which plays an important role in wound healing, was improved by more than two times compared to other similar indirubin derivatives.

That is, it was confirmed that the indirubin derivatives of Examples 1 and 2 exhibited an effect that was not expected at all when compared with the indirubin derivative having a conventional very similar structure, which is a remarkably superior effect beyond the expected effect, that is, It would be said to be an increase in significant effect.

Experimental Example 2. Confirmation of wound healing efficacy and collagen promotion efficacy through animal testing

Using the indirubin derivatives prepared from Examples 1 and 2, the indirubin derivative (IO) of Comparative Example 1, and EGF known as a conventional wound treatment agent as positive controls, the effect of promoting wound healing was confirmed through animal experiments.

The hair on the back of a 7-week-old C3H mouse that entered the resting period was removed, and a 1.5×1.5 cm wound was made on the area where the hair was removed. Vehicle (10% DMSO, 45% ethanol, 18% propylene glycol, 27% water) was applied to the control group, and 20 µl of the indirubin derivative (IO) of Comparative Example 1 was applied once daily for 12 days to the control group. To the positive control group, 100 μM of EGF was applied once daily for 12 days, and the indirubin derivative of Example 2 (labeled A3051) of Example 2 was applied once daily for 12 days in the first experimental group, and Example 1 was applied to the second experimental group. The indirubin derivative (represented by A3334) of 20 μl was applied once daily for 12 days.

The tissues of each experimental mouse were fixed by immersing in 4% paraformaldehyde, embedded in paraffin, and flaked to a thickness of 4 μm. After deparaffinization and rehydration of the sliced tissue, Masson's trichrome staining and picrosirius red staining were performed to evaluate the degree of collagen synthesis. For Masson's trichrome staining, slides were placed in Weigert's Iron Hematoxylin Solution for 10 minutes and Biebrich Scarlet-Acid Fuchsin and Aniline Blue for 5 minutes. In the case of picrosirius red staining, it was placed in Weigert's solution for 8 minutes and picrosirius red for 1 hour.

3 shows the passage of time (day 0, 14) for the mouse acute wound tissue treated with the indirubin derivative of Examples 1 and 2, the indirubin derivative of Comparative Example 1, and EGF (positive control) according to the present invention. It shows the pictures taken according to (1st) and the staining results. In addition, in the lower part of FIG. 3 (collagen staining), a mouse acute wound tissue treated with the indirubin derivative of Example 1 and Example 2, the indirubin derivative of Comparative Example 1, and EGF (positive control) according to the present invention was treated with Masson's trichrome, The results of staining with picrosirius red are shown.

4 shows the passage of time (1, 3, and the indirubin derivative of Example 1 and Example 2, the indirubin derivative of Comparative Example 1, and the mouse acute wound tissue treated with EGF (positive control) according to the present invention. It is a graph showing the quantification of the degree of re-epithelialization measured according to days 5, 9, and 12).

5 is a graph measuring wound size (cm 2) on day 14 in mice treated with the indirubin derivative of Example 1 and Example 2, the indirubin derivative of Comparative Example 1, and EGF (positive control) according to the present invention. .

As shown in FIGS. 3 to 5, when each of the indirubin derivatives prepared from Examples 1 and 2 were treated, it was confirmed that acute wounds healed faster in the mouse model compared to the control group, Comparative Example 1 and the positive control group. That is, as a result of applying the indirubin derivatives of Examples 1 and 2 to the wound of the mouse, the acute wound in the mouse was re-epithelialized equal to or faster than the positive control EGF, and various kinds of collagen tissues were formed. It can be seen that the wound healing markers increase.

In particular, in the case of the indirubin derivative (A3334) of Example 1, it exhibits a slightly better wound healing effect than the positive control EGF. It was also confirmed that the'scar improvement' effect of recovering evenly appeared.

In the case of the indirubin derivative (A3051) of Example 2, it was confirmed that the indirubin derivative (A3051) exhibited a wound healing effect of 2 to 6 times or more at the same time as that of the positive control EGF, through which it can be seen that the wound is recovered more quickly. In addition, it was confirmed that the epidermal layer of the skin recovered cleanly and uniformly so that no traces such as dents or dents were left as the wound healed than when treated with the positive control EGF. It was confirmed that it is also very effective in'improvement'.

In other words, it was confirmed that the indirubin derivatives of Examples 1 and 2, unlike conventional wound treatment agents or indirubin derivatives, not only heal wounds, but also have an unexpected and remarkable effect of improving scars that induce uniform recovery of the epidermal layer of the skin.

In addition, as shown in FIG. 3, Masson's trichrome staining, picrosirius red staining, and van Gieson staining are staining methods for confirming the type and amount of collagen, respectively, and indirubin derivatives prepared from Examples 1 and 2, comparison As a result of confirming the acute wound tissue of mice treated with the indirubin derivative prepared from Example 1 and EGF (positive control) by the above two staining methods, when the indirubin derivatives of Examples 1 and 2 were treated, the wound recovery of cells was also faster. In addition, it can be seen that the amount of collagen was significantly increased compared to the control group, Comparative Example 1, and the positive control group.

As a result of comparing the mouse model prepared by treatment with epithelial growth factor (EGF) in the same manner as the mouse model treated with Example 2 through two staining (Masson's Trichrome, Picrosirius Red), prepared from Examples 1 and 2 It was confirmed that the use of the indirubin derivative exhibited superior wound healing effect compared to the case where the conventional epithelial cell growth factor (EGF) was directly treated and the case where the indirubin derivative of Comparative Example 1 was treated.

Experimental Example 3. Analysis of the wound healing efficacy of a composition containing a complex of an indirubin derivative (Formula 1 or 2) as an active ingredient

In order to analyze the wound healing efficacy, the indirubin derivative prepared from Example 2, the methyl vanylate of Example 3, the hesperidin of Example 4, the quercitrin of Example 5, and the complexes prepared from Examples 7 to 9 were treated. At this time, it was attempted to compare the mobility of cells.

In wound healing, HaCaT cells, that is, human keratinocyte cells, which play an important role in wound healing, were cultured in a 12-well plate for 24 hours. In this case, DMEM containing 10% FBS was used. After 24 hours of incubation, the medium was exchanged with DMEM medium containing 5% FBS, and then a scratch was made in the center of the cells using a 1000p (blue) tip. Thereafter, the indirubin derivative prepared from Example 2, the methyl vanylate of Example 3, the hesperidin of Example 4, the quercitrin of Example 5, and the composites prepared from Examples 7 to 9 were respectively added to the scratched portion. After treatment at a concentration of 1 μM or 5 μM, cell motility was observed after 18 hours.

Specifically, 1 μM of the indirubin derivative prepared from Example 2, the hesperidin of Example 4, and the quercitrin of Example 5 were used, and the methyl vanylate of Example 3 was 5 μM, and Examples 7 to 9 The composite prepared from was prepared by mixing 1 μM indirubin derivative, 5 μM methyl vanylate, 1 μM hesperidin, or 1 μM quercitrin.

Figure 6a is the movement of cells according to the treatment of the indirubin derivative prepared from Example 2, the methyl vanylate of Example 3, the hesperidin of Example 4, the quercitrin of Example 5, and the complexes prepared from Examples 7 to 9 In order to confirm the ability, a photograph showing the mobility of cells in keratinocytes, Figure 6b is the indirubin derivative prepared from Example 2, methyl vanylate of Example 3, hesperidin of Example 4, and Example 5 In order to confirm the mobility of cells according to the treatment of quercitrin and the complexes prepared from Examples 7 to 9, a graph quantitatively showing the cell migration rate in keratinocytes cells. At this time, the control refers to keratinocyte cells that have not been treated with anything. The wound closure rate (%) was determined by the indirubin derivative prepared from Example 2, methyl vanylate of Example 3, hesperidin of Example 4, quercitrin of Example 5, and keratinocytes playing an important role in wound healing. The results (FIG. 6) of each treatment of the composites prepared from Examples 7 to 9 were quantified by cell migration rate.

As shown in Figure 6, when compared with the control cells treated with methyl vanylate of Example 3, hesperidin of Example 4, and quercitrin of Example 5 alone, it was confirmed that there was no difference in the motility of the cells. .

On the other hand, it can be seen that the motility in the cells treated with the complexes prepared from Examples 7 to 9 was improved by up to 2.5 to 4 times or more than the motility in the control cells.

This experiment was measured after 18 hours, which was 6 hours earlier than the previous experiment of FIG. 1, after treating the cells with each active ingredient, compared to when the indirubin derivative A3051 of Example 2 was treated alone on cells. It was confirmed that the treatment of the composites prepared from Examples 7 to 9 improved the motility of cells of keratinocytes, which plays an important role in wound healing, faster and more (1.4 to 2.3 times or more).

That is, compared with the treatment of the indirubin derivative of Example 2 or the methyl vanylate of Example 3, the hesperidin of Example 4, and the quercitrin of Example 5 alone, an effect that was not expected at all (recovery of cellular motility It was confirmed that the reduction of time and recovery of motility (2.5-4 times)) were shown, and this would be a remarkably excellent effect, that is, a significant increase in the effect beyond the expected effect.

Experimental Example 4. Confirmation of the wound healing efficacy of the composite through animal testing

Example 2 Indirubin derivative (0.5 mM), methyl vanillate of Example 3 (2.5 mM, 5 mM), composite prepared from Example 7 (A3051 0.5 mM, Methyl vanillate 3 or 5 mM) and conventional wound treatment agents Known EGF (positive control 1) and PTD+VPA (Arg Lys Thr Gly His Gln Ile Cys Lys Phe Arg Lys Cys, a complex of cell-penetrating peptide and valproic acid, positive control 2) were treated in an animal model. Through this, we tried to confirm the effect of promoting wound healing.

The hair on the back of a 7-week-old C3H mouse that entered the resting period was removed, and a 1.5×1.5 cm wound was made on the area where the hair was removed. EGF 100 μM or PTD+VPA was applied once daily for 12 days to the positive control group, and to the experimental group, Example 2 indirubin derivative (0.5 mM), methyl vanylate (2.5 mM, 5 mM) of Example 3, Example The composite prepared from 7 was applied once daily for 12 days by 20 µl.

The wound size of each tested mouse was measured, and the wound tissue was fixed by immersing in 4% paraformaldehyde, embedded in paraffin, and flaked to a thickness of 4 μm. After deparaffinization and rehydration of the flaked tissue, the degree of re-epithelialization was analyzed.

Figure 7 is an Example 2 indirubin derivative (0.5 mM) according to the present invention, the methyl vanylate of Example 3 (2.5 mM, 5 mM), the composite prepared from Example 7, EGF (positive control 1) and PTD+ The photographs and staining results of the mouse acute wound tissue treated with VPA (positive control 2) are shown according to the passage of time (day 0, day 14). In addition, the lower left graph of FIG. 7 is an example 2 indirubin derivative (0.5 mM) according to the present invention, methyl vanylate (2.5 mM, 5 mM) of Example 3, a composite prepared from Example 7, EGF (positive control) 1) and PTD+VPA (positive control 2) is a graph showing the quantification of the degree of re-epithelialization measured over time (day 0, day 14) for the mouse acute wound tissue treated. The lower right graph of FIG. 7 shows the indirubin derivative (0.5 mM) of Example 2, the methyl vanylate of Example 3 (2.5 mM, 5 mM), the composite prepared from Example 7, EGF (positive control 1) according to the present invention. ) And PTD+VPA (positive control 2) in mice treated with the wound size (cm2) on the first and 14th days. At this time, the control (con) was treated with a solution to which no sample was added, and the control (con1) was applied with water to which nothing was added, and the control (con2) was 10% DMSO, 45% ethanol, and 18% propylene glycol. Cole, 27% water was applied.

Example 2 Indirubin derivative (0.5 mM) is expressed as 0.5 mM A3051, and the group treated with methyl vanillate (2.5 mM, 5 mM) of Example 3 is expressed as methyl vanillate 2.5 mM and 5 mM. The composite prepared from Example 7 is labeled A3051+Methyl vanillate2.5 or 5. On the graph, methyl vanillate was expressed as M (dose).

As shown in FIG. 7, when the indirubin derivative of Example 2 (0.5 mM), the methyl vanylate of Example 3 (2.5 mM, 5 mM), and the composite prepared from Example 7 were treated, respectively, the control group and the positive control group Compared to the mouse model, it was confirmed that the acute wound healed rapidly. The above substances were treated in each experimental animal, and all wounds had the same size on day 1, but after 12 days, mice administered the complex of Example 7 in which 0.5 mM A3051 and 5 mM methyl vanillate were mixed were administered as positive control group 1. It was confirmed that the wound recovery was significantly faster than that of phosphorus EGF, and the scar was also less, and it was confirmed that re-epithelialization was performed equally or faster than that of PTD+VPA, a conventional wound treatment agent.

Among the complexes of Example 7, it was confirmed that the complex mixed with 0.5 mM A3051 and 5 mM Methyl vanillate exhibited a wound recovery effect of 4 times or more at the same time as that of EGF, which is the positive control 1, and through this, it was found that the wound healed more quickly. I can. In addition, it was confirmed that the wound showed twice the wound healing effect than when treated with PTD+VPA, which was the positive control 2, and the epidermal layer of the skin was clean so that no traces such as dents or dents were left while healed. And it was confirmed that it recovered evenly. That is, it was confirmed that the indirubin derivative of Example 2 also had a wound healing effect, but the composite of Example 7 mixed with 0.5 mM A3051 and 5 mM methyl vanillate was most effective in wound healing and scar improvement.

Experimental Example 5. Confirmation of the wound healing efficacy of the compound of Shiva extract and indirubin derivative through animal testing

The indirubin derivative of Example 1 (0.1 μM, 1 μM), the indirubin derivative of Example 2 (0.1 μM, 1 μM), the complex of Examples 10 and 11 (0.1 μM, 1 μM with respect to 1 μM/ml of Shiva extract) Each containing the indirubin derivative of) was treated on the cells. It was to compare its mobility.

Cells that were not treated with anything as a control in this experiment (con), those treated with Shiva extract (1 µg/ml) prepared in Example 6 alone (1 µg/ml) and VPA as a positive control Proic acid) 100 μM was prepared.

The experiment was carried out as follows. First, the HaCaT cells were cultured in a 12-well plate for 24 hours. The HaCaT cells are human keratinocyte cells, which are known to play an important role in the wound healing process. At this time, DMEM containing 10% FBS was used. After 24 hours of incubation was completed, the medium was replaced with DMEM medium containing 5% FBS, and a scratch was made in the center of the cells using a 1000p (blue) tip. The sample was treated on the scratched area and the mobility and motility of the cells were observed after 18 hours.

The samples include the indirubin derivatives of Example 1 (0.1 μM, 1 μM), the indirubin derivatives of Example 2 (0.1 μM, 1 μM), and the complexes of Examples 10 and 11 (0.1 per 1 μg/ml of Schizophrenia extract). µM and 1 µM of indirubin derivatives were used, respectively, and the name of the sample and the administered dose were written in the figure.

Figure 8a shows cells treated with anything as a control (con), one treated with the extract (1 µg/ml) of Shivais prepared from Example 6 alone (1 µg/ml of Shinamu) and VPA (valproic acid) as a positive control. ) 100 μM was treated with keratinocytes, respectively, and 18 hours later, a photograph of the mobility of the cells was taken.

8B shows the indirubin derivatives of Example 1 (0.1 μM, 1 μM), the indirubin derivatives of Example 2 (0.1 μM, 1 μM), and the complexes of Examples 10 and 11 (0.1 μM for 1 μg/ml of Shiva extract) , 1 μM of each of the indirubin derivatives) were treated with keratinocytes, respectively, and 18 hours later, the mobility of the cells was taken. Figure 8c is a graph showing the calculation of the cell migration rate from Figures 8a, b.

As shown in Fig. 8a, when the extract (Example 6) is treated alone, or the indirubin derivative (shown as 0 µg/ml) is treated alone, as shown in Fig. 8B, the control (con) and It can be seen that there is no difference in cell motility. This can also be confirmed in Figure 8c showing the cell migration rate.

On the other hand, in the case of the composites mixed at various ratios prepared from Examples 10 and 11 of the present invention, it was confirmed that the cell motility was remarkably recovered, which was about twice or more than when treated alone.

In particular, looking at the recovered cell migration rate (%) compared to the control group, it was increased by 28% when treated with only Shiunae extract alone, and 50% and 114% when treated with the A3051 indirubin derivative alone, respectively. On the other hand, when the complex of Example 10 was treated, the bar increased by 71% and 246%, which was confirmed to be about 1.5 to 2 times higher than that of the sum of the extract of Schizophrenia and the indirubin derivative. That is, it can be seen that the composition for treating wounds of the present invention has a remarkable effect than that of using the extract and indirubin derivatives respectively, which exceeds the predictable value simply summed up the values obtained from the respective compositions.

Experimental Example 6. Confirmation of the wound healing efficacy of the complex of Shiva extract and indirubin derivatives through animal testing

The indirubin derivative of Example 1 (0.5 mM), the indirubin derivative of Example 2 (0.5 mM), and the complex of Examples 10 and 11 (each containing 0.5 mM of indirubin derivative with respect to 1 mg/ml of Shiva extract) Was treated in an animal model, and through this, the effect of promoting wound healing was attempted to be confirmed.

The hair on the back of a 7-week-old C3H mouse that entered the resting period was removed, and a 1.5×1.5 cm wound was made on the area where the hair was removed. The sample was applied once daily for 12 days to the wound area of the experimental animal. On the 0th and 12th days, the wound size of each experimental animal was measured, and the wound tissue was fixed by immersing in 4% paraformaldehyde, embedded in paraffin, and flaked to a thickness of 4 μm. After deparaffinization and rehydration of the flaked tissue, the degree of re-epithelialization was analyzed and compared. Masson's trichrome staining and picrosirius red staining were performed to evaluate the degree of collagen synthesis. For Masson's trichrome staining, slides were placed in Weigert's Iron Hematoxylin Solution for 10 minutes and Biebrich Scarlet-Acid Fuchsin and Aniline Blue for 5 minutes. In the case of picrosirius red staining, it was placed in Weigert's solution for 8 minutes and picrosirius red for 1 hour. β-catenin, pCNA, keratin 14, and collagen I are important markers related to wound healing. The skin tissue section used in FIG. 9A was fluorescently stained using an antibody of β-catenin, pCNA, keratin 14, and collagen I, and the results were observed through a fluorescence microscope.

Samples were the indirubin derivative of Example 1 (0.5 mM), the indirubin derivative of Example 2 (0.5 mM), and the complex of Examples 10 and 11 (0.5 mM of the indirubin derivative per 1 mg/ml of Shiva extract), respectively. Including), the extract (1 mg/ml) prepared from Example 6 was treated alone (1 mg/ml), or 100 μM of EGF and 500 mM of VPA (valproic acid) were used as positive controls. I did. At this time, the control (con) was treated with a solution to which no sample was added, and the control (con1) was applied with water to which nothing was added, and the control (con2) was 10% DMSO, 45% ethanol, and 18% propylene glycol. Cole, 27% water was applied.

9A shows the indirubin derivative of Example 1 (0.5 mM), the indirubin derivative of Example 2 (0.5 mM), the complex of Examples 10 and 11, the control (con), and the extract of Schizophrenia of Example 6 according to the present invention ( Shinamu), positive control (EGF and VPA) treated mouse acute wound tissue, the photographs taken according to the time (day 0, day 12) and staining results are shown.

9B is a graph showing the size (mm 2) of the mouse acute wound in each group measured on day 0 in FIG. 9A, and FIG. 9C is a graph showing the size (mm 2) of the mouse acute wound in each group measured on the 12th day in FIG. 9A. This is a graph that was measured and shown.

In the drawing, according to the sample treated on the wound area of the experimental animal, the name and capacity of the sample are indicated. For example, Example 2 indirubin derivative (0.5 mM) is expressed as A3051 0.5 mM, and the complex of Example 11 is expressed as A3051 (0.5 mM) + Shrub 1 mg/ml, EGF (100 μM), It is expressed as VPA (500 mM) and the like.

As shown in FIG. 9, in the case of the composites of Examples 10 and 11, when only the indirubin derivative was treated alone, it was confirmed that the degree of wound size recovery was similar to that of the positive control group (EGF, VPA).

However, as shown in FIG. 9, in the case of the composites of Examples 10 and 11, it was confirmed through the naked eye and the wound size that the wound size was recovered the most compared to the control group and the positive control group.

In addition, there was almost no difference in the results of collagen staining by Picrosirius Red and Masson's Trichrome, including β-catenin, pCNA, keratin 14, and collagen I, in the case of treatment with only the extract of Schizophrenia of Example 6 and the control group (con). On the other hand, when the indirubin derivatives of Examples 1 and 2 were treated alone, β-catenin, pCNA, keratin 14, and collagen I expression were confirmed than the control (con), but it was confirmed that it was slightly lower than the positive control.

However, the acute wound tissue treated with the complexes of Examples 10 and 11 in which the indirubin derivative of the same concentration was mixed with the extract of Schizophrenia had significantly increased β-catenin, pCNA, keratin 14, and collagen I expression compared to the positive control group. It was confirmed that the result of collagen staining by Picrosirius Red and Masson's Trichrome was also remarkably excellent. In other words, the complexes of Examples 10 and 11 were clearer than the positive control group so that the expression of β-catenin, pCNA, keratin 14, and collagen I was increased, and no traces such as healing or dents were left. It was confirmed that it recovered evenly.

The complexes of Examples 10 and 11 were mixed with the extracts of Shiva tree, which did not show any effect on the expression of β-catenin, pCNA, keratin 14, and collagen I, and the indirubin derivatives of Examples 1 and 2 having some effect, scar/wound It was confirmed that the expression of various factors having an effect on healing was improved, which did not show an effect by itself, but when used in combination with an indirubin derivative, it is believed to promote the effect.

Experimental Example 7. Measurement of the solubility of the active ingredient of the present invention

In order to select an emulsion solution, the solubility of the indirubin derivative of Example 2 was measured in various oils and surfactants. A sufficient amount of the indirubin derivative powder prepared in Example 2 was added to each oil or surfactant (1 ml), followed by vortexing for 30 minutes. Then, in order to reach equilibrium, the supernatant was shaken for 72 hours at 50 rpm in a culture tank at 37° C., and then centrifuged at 16,100 × g for 5 minutes. The supernatant was appropriately diluted with methanol and analyzed by LC/MS/MS. The types of oil or surfactant and the measured solubility are shown in Table 1 below.

As the analysis equipment, Waters xevo TQ MS-ACQUITY UPLC System was used. The column was analyzed at room temperature using ACQUITY UPLCㄾ BEH (C18, 1.7 μm, 2.1×50 mm). The mobile phase was used after filtering deionized distilled water containing 0.1% formic acid and acetonitrile (30:70, v/v) containing 0.1% formic acid through a 0.2 μm membrane filter. The flow rate was 0.25 ml/min and 0.5 μl of the sample was injected and analyzed. LC/MS/MS conditions for Decursin analysis were 38 V for Cone and 32 V for Collison, and the mother and daughter ions were monitored with m/z values of 329 and 229, respectively.

Kinds Kinds Solubility
(mg/ml) oil Olive oil
Olive oil 0.83 Sunflower oil
Sunflower oil 1.35 Kolliphor EL 5.4 Surfactants Tween 20 4.58 Tween 80 4.92 Co-surfactant
(Co-surfactant) Transcutol P 1.15 Propylene glycol
Propylene glycol 0.25 PEG 400 3.1

10 is a photograph of an emulsion solution prepared by dissolving the indirubin derivative prepared from Example 2 of the present invention in the various oils or surfactants, respectively, as shown in FIG. 10 and Table 1, Example 2 according to the present invention. The indirubin derivative prepared from Kolliphor® EL, Tween 20, Tween 80, and PEG 400, which have the highest solubility among oily solvents, was first established to form an emulsion solution. The Kolliphore EL, Tween 20 or Tween 80, PEG 400 is a stable substance used in the composition of various formulations including skin administration because it is harmless and safe to the human body.

Experimental Example 8. Ternary diagram analysis

Based on the solubility test results, Kolliphorㄾ EL was selected as the oil phase of the emulsion solution, and a mixture of Tween 80 and PEG 400 was selected as a surfactant. The ternary phase diagram was completed using the _{H 2} O titration method at room temperature in order to check the area where the emulsion is formed with the composition of the emulsion solution selected in this way. As a surfactant, a surfactant mixture was prepared by mixing Tween 80 and PEG 400 in a ratio of 1:1, 2:1, and 1:2, and this was prepared in various ratios (0.5:9.5, 1:9) with the oily Kolliphor® EL. , 2:8, 3:7, 4:6, 5:5, 6:4, 7:3, 8:2, 9:1, 9.5:0.5) to prepare an emulsion solution. While stirring the emulsion solution, an area that maintains uniformly mixed state when observed with the naked eye while dropping water at a rate of 1 ml/min using an infusion pump is indicated as an emulsion on the phase diagram, and the three-way diagram It is shown in Figs. 11A, 11B and 11C.

11A is a ternary phase diagram for an emulsion solution prepared with a surfactant in which Tween 80 and PEG 400 are mixed in a ratio of 2:1, and FIG. 11B is an interface in which Tween 80 and PEG 400 are mixed in a ratio of 1:1. It is a ternary phase diagram for an emulsion solution prepared as an active agent, and FIG. 11C is a ternary phase diagram for an emulsion solution prepared as a surfactant in which Tween 80 and PEG 400 are mixed in a ratio of 1:2. In the ternary phase diagram of FIG. 9, regions forming a stable emulsion are indicated by red dots.

11a, b, c, it was confirmed that the emulsion solution using a surfactant in which Tween 80 and PEG 400 were mixed in a weight ratio of 1:2 had the widest numerical range, and this was selected. Thereafter, in order to finally select the ratio of the surfactant and the oil phase, the solubility of the emulsion in the emulsion solution prepared in various ratios, droplet size, viscosity, and zeta potential were compared.

Experimental example 9. Formulation development of pharmaceutical composition for wound prevention or treatment

An attempt was made to develop an emulsion formulation of an oil-in-water type that allows the indirubin derivative prepared in Example 2 to have a certain solubility and long-term stability, and for this purpose, a surfactant, polyethylene glycol, and oil were mixed to select the most stable emulsion solution.

An emulsion solution was prepared with various compositions of the surfactant selected from Experimental Example 5, polyethylene glycol, and oil 10:30:60 to 90:3:7. In order to prepare an emulsion containing the active ingredient of the present invention, the active ingredient (indirubin derivative powder of Example 2) was added to the emulsion solution to a concentration of 10% by weight with respect to the total weight of the total composition, and stirred overnight, to prepare the emulsion. (For example, if the weight of the final composition is 100 g, 20 g of the indirubin derivative powder prepared in Example 2 was mixed with 80 g of the emulsion solution).

However, the composition may further include a solubilizing agent, cyclodextrin, and in this experiment, in order to increase the solubilization of the indirubin derivative, 100 parts by weight of the indirubin derivative (active ingredient), based on 100 parts by weight of cyclodextrin. What added parts by weight was used.

The prepared emulsion formulation was an opaque red color uniformly mixed with the naked eye, and the solubility, droplet size, viscosity, and zeta potential of each of the emulsion formulations were analyzed and shown in Table 2.

division Emulsion solution
(Kolliporeㄾ EL:Tween 80:PEG400)
Weight ratio Solubility
Solubility
(mg/ml) Droplet size (nm) viscosity
Viscosity (mPas) Zetapotential (mV) F1 10:30:60 2.99 1525.7 23.33 -5.14 F2 20:26:54 3.58 921.3 44.46 -6.87 F3 30:23:47 3.81 624.9 59.12 -7.23 F4 40:20:40 4.11 479.4 65.07 -6.94 F5 50:16:34 5.67 332.3 72.72 -7.99 F6 60:13:27 6.86 264.9 88.85 -9.72 F7 70:10:20 7.59 183.8 91.39 -10.83 F8 80:6:14 7.69 41.5 98.3 -20.38 F9 90:3:7 6.88 29.3 99.52 -20.99

As shown in Table 2, the size of the droplets decreased as the content of oil increased, and it was confirmed that all of them have appropriate physical properties for use as pharmaceutical and cosmetic compositions, but the most desirable size, viscosity, zeta potential, and solubility were determined. It can be seen that what has it is F8.

Experimental example 10. emulsion Formulation dependent stability evaluation-1

In order to check the stability of the prepared emulsion formulation, emulsion formulations F8 and F10-12 were prepared with the composition in Table 3 below through the same manufacturing process as in Experimental Example 6. The stability of the emulsion formulation was measured based on the relative solubility% change at low temperature (4 °C) and room temperature (25 °C). The relative solubility% from immediately after manufacture to 3 months was recorded in FIGS.

division Emulsion composition Active ingredient
Indirubin derivative of Example 2 Solubilizer
(2-hydroxy-cyclodextrin) Awards oil Surfactants Polyethylene glycol DMSO
Control - - - 12.5 g 12.5 g Distilled water
100 g F8 Kollipore® EL
80 g Tween 80
6.7 g PEG400
13.3 g 12.5 g 12.5 g Distilled water
100 g F10 - - PEG400
100 g 12.5 g 12.5 g Distilled water
100 g F11 - Tween 80
100 g - 12.5 g 12.5 g Distilled water
100 g F12 Kollipore® EL
100 g - - 12.5 g 12.5 g Distilled water
100 g

Figure 12 is a graph showing the relative solubility% change of F8, F10 ~ 12 emulsion formulations measured at room temperature (25 °C), Figure 13 is a relative solubility% of F8, F10 ~ 12 emulsion formulations at low temperature (4 °C) It is a graph showing the change measured.

As shown in Figs. 12, 13, and 3, as a result of examining the relative solubility change after standing at low temperature and room temperature for 3 months, in the case of the emulsion (F8) of the present invention, the solubility% change immediately after preparation and after 3 months On the other hand, in the case of the emulsion (F12, 11, 10) prepared with an emulsion composition different from the present invention, the solubility change occurred more than 40% at low temperature and room temperature conditions immediately after manufacture and after 3 months. It was confirmed that the solubility was rapidly lowered by more than 80%.

Experimental example 11. emulsion Formulation Dependent Stability Evaluation-2

In order to check whether the active ingredient of the indirubin derivative of Example 2 stably maintains its activity in the emulsion formulation, F8, F10 to 12 emulsion formulations were prepared with the composition in Table 3 through the same manufacturing process as in Experimental Example 7. I did. The stability against the activity of the indirubin derivative in the emulsion formulation was measured based on the relative wnt reporter activity% change at low temperature (4 °C) and room temperature (25 °C). The relative wnt reporter activity% from immediately after manufacture to 3 months was recorded in FIGS. 14 and 15.

The wnt reporter activity% was measured by the following method. The wnt reportter activity was measured 24 hours after treatment of the active ingredient prepared as an emulsion formulation in the HEK TOP reporter cell line.

14 is a graph showing changes in relative wint reporter activity% of F8, F10~12 emulsion formulations at room temperature (25°C), and FIG. This is a graph showing the change in wnt reporter activity%.

As shown in FIGS. 14 and 15, when left at low temperature and room temperature for 3 months, the relative wnt reporter activity% change was examined. In the case of the emulsion (F8) of the present invention, wnt reporter activity immediately after manufacture and after 3 months While the% change was not large, in the case of the emulsion (F12, 11, 10) prepared with an emulsion composition different from the present invention, the wnt reporter activity was reduced by more than 40% at low and room temperature conditions immediately after manufacture and after 3 months. It was confirmed that the solubility was rapidly lowered by more than 80% under low temperature storage conditions.

Formulation Example 1: Cream-type cosmetic composition for preventing or treating wounds

In this preparation example, it was intended to prepare a cream-type cosmetic composition for preventing or treating wounds containing a complex mixture of the indirubin derivative of Formula 1 or Formula 2 as an active ingredient. The ingredients containing the indirubin derivative of Examples 1 or 2 were mixed according to the following contents to prepare a cream-shaped cosmetic composition.

4.0% by weight of the indirubin derivative of Example 1 or 2

2.0% by weight of glycerin

0.2% by weight of paraben

Allantoin 2.0% by weight

Betaine 3.0% by weight

Sodium hyaluronate 1.0% by weight

Tocopherol acetate 5.0% by weight

Shea butter 2.0% by weight

1.0% by weight of trehalose

Weight% of appropriate amount of preservative and fragrance

Purified water to 100% by weight

100% by weight in total

Formulation Example 2: A pharmaceutical composition (ointment) for preventing or treating wounds

In this preparation example, a pharmaceutical composition (ointment) for preventing or treating wounds containing the indirubin derivative of Formula 1 or 2 as an active ingredient was intended to be prepared. Components containing the indirubin derivative of Examples 1 or 2 were mixed according to the following contents to prepare a pharmaceutical composition (ointment) for preventing or treating wounds.

10% by weight of the indirubin derivative of Example 1 or 2

8% by weight of diethyl sebacate

5% by weight brazing

Polyoxyethylene oleyl ether phosphate 6% by weight

Weight% of appropriate amount of sodium benzoate

Vaseline to 100% by weight

100% by weight in total

Formulation Example 3: Application to milk

99.9% by weight of milk

0.1% by weight of the indirubin derivative of Example 1 or 2

Formulation Example 4: Preparation of beverage

Indirubin derivative of Example 1 or 2 10 mg

Calcium lactate 50 mg

Citric acid 5 mg

Nicotinic acid amide 10 mg

Sodium riboflavin hydrochloride 3 mg

Pyridoxine hydrochloride 2 mg

10 mg arginine

Sucrose fatty acid ester 10 mg

200 ml of water

Claims (40)

Including an indirubin derivative represented by the following Formula 1 or Formula 2 as an active ingredient,
A pharmaceutical composition for the prevention or treatment of wounds, characterized in that the active ingredient does not contain the extract.
[Formula 1]

[Formula 2]

delete The method of claim 1,
The composition is a pharmaceutical composition for preventing or treating wounds, characterized in that it further comprises any one or more selected from the group consisting of methyl vanylate, hesperidine and quercitrin.
The method of claim 1,
The wound is a wound prevention or treatment pharmaceutical composition, characterized in that the injury is caused by burns, ulcers, trauma, post-surgical, childbirth, chronic wounds or dermatitis.
The method of claim 1,
The wound prevention or treatment is a pharmaceutical composition for preventing or treating wounds, characterized in that due to promotion of the movement speed of keratinocytes, activation of the movement and proliferation of fibroblasts, and promotion of collagen production.
The method of claim 1,
The pharmaceutical composition is a pharmaceutical composition for preventing or treating wounds, characterized in that for topical application to the skin.
The method of claim 1,
The pharmaceutical composition is formulated in any one formulation selected from the group consisting of creams, gels, ointments, emulsions, suspensions, sprays, and transdermal pitch agents.
The method of claim 1,
The pharmaceutical composition is a pharmaceutical composition for preventing or treating wounds, characterized in that the emulsion formulation further comprises an oil, a surfactant, and polyethylene glycol.
The method of claim 8,
A pharmaceutical composition for preventing or treating wounds, characterized in that the mixing weight ratio of the oil, surfactant and polyethylene glycol is 0.3-30:1:2-2.5.
The method of claim 1,
The active ingredient is a pharmaceutical composition for preventing or treating wounds, characterized in that it contains 1 to 20% by weight based on the total weight of the composition.
The method of claim 8,
The surfactant is polyoxyethylene sorbitan monolaurate (Tween 20), polyoxyethylene sorbitan monopalmitate (Tween 40), polyoxyethylene sorbitan monostearate (polyoxyethylene sorbitan monostearate). , Tween 60) and polyoxyethylene sorbitan oleate (polyoxyethylene sorbitan oleate, Tween 80). A pharmaceutical composition for preventing or treating wounds, characterized in that at least one selected from the group consisting of.
The method of claim 8,
The oil is a pharmaceutical composition for preventing or treating wounds, characterized in that at least one selected from the group consisting of polyethoxylated castor oil (KolliphorEL), sunflower oil, and olive oil.
The method of claim 8,
The composition further comprises cyclodextrin, wherein the composition contains 100 to 1000 parts by weight of cyclodextrin based on 100 parts by weight of the active ingredient present in the composition.
The method of claim 8,
The emulsion formulation is a pharmaceutical composition for preventing or treating wounds, characterized in that it is a stable formulation that does not show a change in wnt activity and solubility in distilled water under a temperature of 25 to 4°C when measured at 3 months.
An indirubin derivative represented by the following formula (1) or formula (2) is used as an active ingredient,
A cosmetic composition for preventing or improving wounds, characterized in that the active ingredient does not contain the extract of Shiva.
[Formula 1]

[Formula 2]

delete The method of claim 15,
The composition is a cosmetic composition for preventing or improving wounds, characterized in that it further comprises any one or more selected from the group consisting of methyl vanylate, hesperidin and quercitrin.
The method of claim 15,
The cosmetic composition is a cosmetic composition for preventing or improving wounds, characterized in that any one selected from the group consisting of lotion, essence, lotion, cream, pack, gel, powder, foundation, and detergent.
The method of claim 15,
The cosmetic composition is a cosmetic composition for preventing or improving wounds, characterized in that the emulsion formulation further comprises oil, surfactant and polyethylene glycol.
The method of claim 19,
A cosmetic composition for preventing or improving wounds, characterized in that the mixing weight ratio of the oil, surfactant and polyethylene glycol is 0.3-30:1:2-2.5.
The method of claim 15,
The active ingredient is a cosmetic composition for preventing or improving wounds, characterized in that it contains 1 to 20% by weight based on the total weight of the composition.
The method of claim 19,
The surfactant is polyoxyethylene sorbitan monolaurate (Tween 20), polyoxyethylene sorbitan monopalmitate (Tween 40), polyoxyethylene sorbitan monostearate (polyoxyethylene sorbitan monostearate). , Tween 60) and polyoxyethylene sorbitan oleate (polyoxyethylene sorbitan oleate, Tween 80) any one or more selected from the group consisting of a cosmetic composition for preventing or improving wounds.
The method of claim 19,
The oil is a cosmetic composition for preventing or improving wounds, characterized in that at least one selected from the group consisting of polyethoxylated castor oil (Kolliphor ㄾEL), sunflower oil and olive oil.
The method of claim 19,
The composition further comprises cyclodextrin, but a cosmetic composition for preventing or improving wounds, characterized in that 100 to 1000 parts by weight of cyclodextrin is included based on 100 parts by weight of an active ingredient present in the composition.
The method of claim 19,
The emulsion formulation is a cosmetic composition for preventing or improving wounds, characterized in that it is a stable formulation that does not show changes in wnt activity and solubility in distilled water under conditions of 25 to 4°C at a temperature as measured at 3 months.
Including an indirubin derivative represented by the following Formula 1 or Formula 2 as an active ingredient,
A food composition for preventing or improving wounds, characterized in that the active ingredient does not contain the extract of Shiva.
[Formula 1]

[Formula 2]

delete The method of claim 26,
The composition is a food composition for preventing or improving wounds, characterized in that it further comprises any one or more selected from the group consisting of methyl vanylate, hesperidin and quercitrin.
An indirubin derivative represented by the following formula (1) or formula (2) is used as an active ingredient,
The composition for skin regeneration and elasticity improvement, characterized in that the active ingredient does not contain the extract.
[Formula 1]

[Formula 2]

delete The method of claim 29,
The composition is a composition for improving skin regeneration and elasticity, characterized in that it further comprises any one or more selected from the group consisting of methyl vanylate, hesperidin and quercitrin.
The method of claim 29,
The indirubin derivative is a composition for improving skin regeneration and elasticity, characterized in that to promote collagen synthesis to promote skin regeneration and improve skin elasticity.
The method of claim 29,
The composition is a composition for skin regeneration and elasticity improvement, characterized in that it is a formulation of a lotion, essence, lotion, cream, pack, gel, powder, foundation or detergent.
The method of claim 29,
The composition is a composition for skin regeneration and elasticity improvement, characterized in that the emulsion formulation further comprises an oil, a surfactant and polyethylene glycol.
The method of claim 34,
The composition for skin regeneration and elasticity improvement, characterized in that the mixing weight ratio of the oil, surfactant and polyethylene glycol is 0.3-30:1:2-2.5.
The method of claim 29,
The active ingredient is a composition for skin regeneration and elasticity improvement, characterized in that it contains 1 to 20% by weight based on the total weight of the composition.
The method of claim 34,
The surfactant is polyoxyethylene sorbitan monolaurate (Tween 20), polyoxyethylene sorbitan monopalmitate (Tween 40), polyoxyethylene sorbitan monostearate (polyoxyethylene sorbitan monostearate). , Tween 60) and polyoxyethylene sorbitan oleate (polyoxyethylene sorbitan oleate, Tween 80), characterized in that any one or more selected from the group consisting of skin regeneration and elasticity improvement composition.
The method of claim 34,
The oil is a composition for skin regeneration and elasticity improvement, characterized in that at least one selected from the group consisting of polyethoxylated castor oil (Kolliphor ㄾEL), sunflower oil, and olive oil.
The method of claim 34,
The composition further comprises cyclodextrin, wherein 100 to 1000 parts by weight of cyclodextrin is included based on 100 parts by weight of an active ingredient present in the composition.
The method of claim 34,
The emulsion formulation is a composition for skin regeneration and elasticity improvement, characterized in that it is a stable formulation that does not show a change in wnt activity and solubility in distilled water under a temperature of 25 to 4°C when measured at 3 months.

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