KR102620622B1 - Cosmetic composition containing EGF with improved storage stability. - Google Patents

Abstract

The present invention provides a cosmetic composition for stabilizing EGF that can secure the stability of EGF by suppressing structural modification, decomposition, and loss of biological activity of EGF due to the external environment, and specifically, can maintain the biological activity of EGF even in a high temperature environment. The purpose is to

Description

Cosmetic composition containing EGF with improved storage stability. {Cosmetic composition containing EGF with improved storage stability.}

The present invention relates to a cosmetic composition for stabilizing EGF with improved storage stability of EGF.

Epidermal growth factor (EGF) is a natural wound healing substance that exists in the human body. When a wound occurs on the skin, etc., it is supplied to cells through blood, sweat, and saliva and acts to heal the wound naturally without scarring. EGF was first isolated from the submandibular gland of rats by Dr. Stanley Cohen in the United States, and when this substance was treated on the back of newborn rats, it was experimentally proven that the eyelids opened faster than those of normal rats, and was described as a 'factor that promotes the growth of epithelial cells.' It was named Epidermal Growth Factor (EGF).

EGF is a polypeptide that structurally has three disulfide bonds and is a factor that promotes the growth of epithelial cells. It transmits signals through the EGF receptor on the cell membrane, thereby promoting the growth and division of mammalian cells, especially epithelial and skin cells. It has been found to induce and promote the growth of epithelial cells. In addition, EGF is known to play a very important role in regulating the molecular level in processes such as wound healing. EGF is present in high concentrations in saliva, urine, breast milk, tears, and blood, and is supplied from the blood when a wound occurs and acts to heal the wound without scarring. In addition, along with FSH (Follicle stimulating hormone), which promotes the secretion of female hormones, it matures fertilized eggs in the uterus and helps regenerate the cornea, which is prone to deterioration due to the lack of blood vessels.

In addition, EGF plays a key role in skin regeneration by promoting cell proliferation not only in epithelial cells but also in fibroblasts that synthesize collagen, a component of the dermis. The human body autonomously regulates the proper supply of EGF to damaged tissues so that the damaged area recovers normally, but if EGF is not supplied properly, it must be supplied from outside to ensure normal recovery. In this respect, EGF can be used in various ways in the field of cosmetics development.

However, EGF reacts sensitively to temperature conditions and has characteristics that do not have high stability in the external environment, so it is emerging as an essential problem that must be solved for commercial use of EGF. In the past, in order to increase the in vivo transmissibility and persistence of EGF, there have been cases in which modified derivatives of the EGF protein were manufactured or bound to the protein transport domain to penetrate into cells in the form of a fusion protein. However, there were problems such as lack of smooth activity or poor expression after being actually introduced into cells.

Accordingly, instead of improving the EGF protein itself or manufacturing it in the form of a fusion protein, there is a need for a method that can increase skin permeability and improve stability by adjusting the ingredients of the cosmetic composition.

The present invention provides a cosmetic composition for stabilizing EGF that can secure the stability of EGF by suppressing structural modification, decomposition, and loss of biological activity of EGF due to the external environment, and specifically, can maintain the biological activity of EGF even in a high temperature environment. The purpose is to

The present disclosure provides a cosmetic composition for stabilizing EGF containing EGF and ectoin.

The cosmetic composition for stabilizing EGF may further include a cyclodextrin dispersion.

The weight ratio of EGF and ectoin may be 1:10 to 1:100.

The weight ratio of EGF and cyclodextrin may be 1:1 to 1:20.

The cyclodextrin dispersion may contain polyol as a solvent.

The weight ratio of polyol and water contained in the cosmetic composition for stabilizing EGF may be 1:1 to 1:10.

The polyol includes one or more selected from the group consisting of glycerin, polyethylene glycol, butylene glycol, pentylene glycol, isopentyldiol, hexanediol, methylpropanediol, sorbitol, erythritol, inositol, maltitol, and xylitol. It may be.

The cosmetic composition for stabilizing EGF includes the steps of preparing an EGF aqueous solution; Mixing the ectoin aqueous solution with the EGF aqueous solution; and optionally mixing a cyclodextrin dispersion into the EGF aqueous solution; It may be manufactured according to a method comprising:

The step of mixing the cyclodextrin dispersion with the EGF aqueous solution may include solubilizing the cyclodextrin using a polyol solvent.

The cosmetic composition for stabilizing EGF may be in the form of lotion, skin, lotion, nutritional lotion, nutritional cream, massage cream, essence, pack, or microneedle patch.

The cosmetic composition for stabilizing EGF according to an embodiment of the present invention can improve the storage stability of EGF.

Specifically, the cosmetic composition for stabilizing EGF according to an embodiment of the present invention can maintain the biological activity of EGF by preventing deformation of EGF at high temperatures.

Figure 1 shows the results of separating proteins from each sample using SDS-page gel electrophoresis.

Hereinafter, the present invention will be described in detail. Terms used in this specification, if not separately defined, should be interpreted as generally understood by those skilled in the art. The drawings and examples of this specification are intended to enable those skilled in the art to easily understand and practice the present invention. Content that may obscure the gist of the invention may be omitted from the drawings and examples, and the present invention is limited to the drawings and examples. It doesn't work.

As used herein, the singular forms “a,” “an,” and “the” are intended to also include the plural forms, unless the context clearly dictates otherwise.

In addition, the numerical range used in the present invention includes the lower limit and upper limit and all values within the range, increments logically derived from the shape and width of the defined range, all double-limited values, and numerical ranges defined in different forms. Includes all possible combinations of upper and lower limits. Unless otherwise specified in the specification of the present invention, values outside the numerical range that may occur due to experimental error or rounding of values are also included in the defined numerical range.

In this specification, terms such as include, have, have, etc. mean the presence of features or components described in the specification, and, unless specifically limited, indicate the possibility of adding one or more other features or components. It is not excluded in advance.

The present disclosure provides a cosmetic composition for stabilizing EGF containing EGF and ectoin. The cosmetic composition for stabilizing EGF can protect EGF from structural deformation, decomposition, and loss of biological activity, and specifically, can maintain the biological activity of EGF even under high temperature conditions of around 40°C.

EGF (Epidermal Growth Factor) is a polypeptide that structurally has three disulfide bonds and is a factor that promotes the growth of epidermal cells and the proliferation of fibroblasts that synthesize collagen. It is included in cosmetic compositions. It is being used usefully. However, EGF has a problem of low stability due to external environmental changes such as temperature conditions, which reduces its industrial applicability. In fact, it has been reported that when a composition containing EGF is cultured at 37°C for 3 hours, the activity of EGF is damaged by 60%. Accordingly, the present disclosure provides a cosmetic composition for stabilizing EGF that can maintain the structural stability of EGF even under high temperature conditions and further maintain the biological activities of EGF, such as collagen synthesis, cell growth, and improvement of skin condition.

The cosmetic composition for stabilizing EGF can maintain the stability of EGF in the post-addition process at around 40°C, which is essential for the cosmetics manufacturing process, and is generally maintained under high-temperature life conditions during the cosmetics quality assurance period of about 1 to 3 years from manufacturing to use. It can maintain the intrinsic activity of EGF, making it easy for mass production and commercial use.

Ectoin is a self-protective substance discovered in halophilic bacteria and is an osmotic substance that minimizes denaturation to survive in extreme conditions. It is a low molecular weight compound and can maintain cell stability and minimize denaturation of biopolymers by hydrating the surrounding area with high water binding capacity. The cosmetic composition for stabilizing EGF of the present disclosure contains ectoin and can prevent decline in EGF activity at temperatures higher than room temperature, for example, up to 40°C or 50°C, or at temperatures lower than room temperature.

According to one embodiment, the weight ratio of EGF and ectoine may be 1:10 to 1:100, specifically 1:20 to 1:80, and more specifically 1:30 to 1:60.

According to one embodiment, the cosmetic composition for stabilizing EGF of the present disclosure may further include a cyclodextrin dispersion, and specifically, the cyclodextrin may be beta cyclodextrin. The cyclodextrin dispersion may produce a synergistic effect and exhibit a more improved EGF stabilizing effect than when ectoin is included alone as a stabilizing material in the cosmetic composition for stabilizing EGF of the present disclosure.

Cyclodextrin is a natural substance obtained from starch, and glucopyranose units form a ring structure through α-(1,4) bonds. The hydroxy groups of each connected glucopyranose are located on the outside of the ring, so the outside of the ring has hydrophilic properties, and the inside of the ring has hydrophobic properties. Cyclodextrin uses the hydrophobic properties of the inside of the ring to incorporate hydrophobic molecules into the inside of the ring to form an inclusion complex. Cyclodextrins can interact with internal hydrophobic molecules and have the effect of altering their properties and stability.

As the cyclodextrin dispersion is added to the cosmetic composition for stabilizing EGF, EGF may be partially incorporated into the ring of cyclodextrin or may be bound to the hydrophilic region outside the ring through electrostatic interaction, but is not limited thereto.

The cyclodextrin dispersion may contain polyol as a solvent. Cyclodextrin can be effectively dissolved in the cosmetic composition for stabilizing EGF through a polyol solvent, and thus can stabilize EGF by interacting with EGF in an aqueous solution.

The polyol may be one or more selected from the group consisting of glycerin, polyethylene glycol, butylene glycol, pentylene glycol, isopentyldiol, hexanediol, methylpropanediol, sorbitol, erythritol, inositol, maltitol, and xylitol. , specifically, it may be glycerin, glycol, isopentyldiol, hexanediol, and methylpropanediol, and more specifically may be glycerin or isopentyldiol, but is not limited thereto.

The weight ratio of EGF and cyclodextrin may be 1:1 to 1:20, specifically 1:5 to 1:15, and more specifically 1:7 to 1:12.

The weight ratio of polyol and water contained in the cosmetic composition for stabilizing EGF may be 1:1 to 1:10, specifically 1:1 to 1:8, and more specifically 1:1 to 1:5.

According to one embodiment, the cosmetic composition for stabilizing EGF of the present disclosure includes the steps of preparing an EGF aqueous solution; Mixing the ectoin aqueous solution with the EGF aqueous solution; and optionally mixing a cyclodextrin dispersion into the EGF aqueous solution; It may be manufactured according to a method comprising:

The cosmetic composition for stabilizing EGF stabilizes the tertiary structure of EGF, which can be deformed due to changes in the external environment, by sequentially mixing the above materials in an EGF aqueous solution as described above. Specifically, the structural stability of EGF at high temperature is stabilized. It can be improved.

The cyclodextrin dispersion may be one in which cyclodextrin is solubilized in a polyol solvent. When cyclodextrin is solubilized in polyol, the difference between the hydrophilicity and hydrophobicity of the pore and the outside is not significant, but as it is mixed with the EGF aqueous solution, the cavity of cyclodextrin in the water phase becomes more hydrophobic, and the hydrophobized cavity becomes more hydrophobic than that of EGF. It is understood that it interacts with the hydrophobic motif to improve the stability of EGF in the aqueous phase.

The cosmetic composition for stabilizing EGF may be in the form of a lotion, skin, lotion, nutritional lotion, nutritional cream, massage cream, essence, pack, or microneedle patch, and may be specifically used as a softening lotion, astringent lotion, nourishing lotion, nourishing cream, and massage. Cream, eye cream, eye essence, essence, cleansing cream, cleansing lotion, cleansing foam, cleansing water, mask pack, powder, body lotion, body cream, body essence, body cleanser, sunscreen cream, hair dye, shampoo, conditioner, toothpaste. , it may have one or two or more formulations selected from mouthwash, hairdresser, hair straightener, lotion, ointment, gel, cream, patch, and spray.

When the formulation of the cosmetic composition for stabilizing EGF is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tragacantha, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc and oxidation are used as carrier ingredients. One or two or more selected from zinc may further be used.

When the formulation of the cosmetic composition for stabilizing EGF is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder may be further used as a carrier ingredient. In particular, in the case of a spray, chlorofluorocarbons may be additionally used. May contain propellants such as lohydrocarbon, propane/butane or dimethyl ether.

When the formulation of the cosmetic composition for stabilizing EGF is a solution or emulsion, a solvent, solubilizing agent, or emulsifying agent may be further used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, and benzyl. These may include benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic esters, polyethylene glycol, or fatty acid esters of sorbitan.

When the formulation of the cosmetic composition for stabilizing EGF is a suspension, the carrier component includes a liquid diluent such as water, ethanol, or propylene glycol; Suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester or polyoxyethylene sorbitan ester may be further used, and crystalline cellulose, aluminum metahydroxide, bentonite, agar or tragacantha may also be added. It can be used as.

When the formulation of the cosmetic composition for stabilizing EGF is a surfactant-containing cleansing agent, the carrier ingredients include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, and sarcosylate. Nate, fatty acid amide ether sulfate, alkylamidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester can be used.

The cosmetic composition for stabilizing EGF according to an embodiment of the present disclosure may further include additional additives depending on the purpose, such as stabilizers, emulsifiers, thickeners, moisturizers, liquid crystal film strengtheners, pH adjusters, One or two or more water-based additives selected from pH adjusters, water-soluble polymers, coating agents, metal ion sequestering agents, organic amines, polymer emulsions, skin nutrients, antioxidants, antioxidant aids, preservatives, fragrances, etc.; and one or two or more oil-based additives selected from fats and oils, waxes, hydrocarbon oils, higher fatty acid oils, higher alcohols, synthetic ester oils, silicone oils, etc.;

Hereinafter, the cosmetic composition for stabilizing EGF according to the present invention will be described in more detail through specific examples. However, the following examples are only a reference for explaining the present invention in detail, and the present invention is not limited thereto, and may be implemented in various forms. Additionally, the terms used in the description in the present invention are only intended to effectively describe specific embodiments and are not intended to limit the present invention.

[Example 1]

An EGF aqueous solution was prepared by dissolving 5 mg of EGF in 5 g of distilled water. 250 mg of ectoine dissolved in 2.5 g of distilled water was added to the EGF aqueous solution, 2.5 g of isopentyldiol was added, and mixed for 10 minutes.

[Example 2]

An EGF aqueous solution was prepared by dissolving 5 mg of EGF in 5 g of distilled water. An ectoine aqueous solution in which 250 mg of ectoine was dissolved in 2.5 g of distilled water was added, and a cyclodextrin dispersion in which 50 mg of beta cyclodextrin was dispersed in 2.5 g of isopentyldiol was added to the EGF aqueous solution and mixed for 10 minutes.

[Comparative Example 1]

5 mg of EGF was added to 7.5 g of distilled water and 2.5 g of isopentyldiol and mixed for 10 minutes.

[Evaluation Example 1] Stability evaluation

The solutions of Examples 1 and 2 and Comparative Example 1 were placed in a 50°C incubator and cultured for 48 hours. The solution of Comparative Example 1 stored at 4°C before culturing was used as a control, and the solutions of Examples 1, 2 and Comparative Example 1 after culturing were analyzed by 15% SDS-PAGE gel electrophoresis. The results are shown in Figure 1. From the results in Figure 1, an undenatured EGF protein band can be confirmed at about 6.2 kDa in each solution, and the concentration of EGF protein measured through each protein band using the Image J program (NIH.GOV) is compared to the control group. Table 1 shows the concentration.

According to Figure 1 and Table 1, when comparing the protein concentration before and after culturing at a high temperature of 50°C, Examples 1 and 2 had an EGF denaturation rate of 10% compared to the Comparative Example, which had an EGF denaturation rate of over 50%. You can see that it is within. In addition, in the case of Example 2 containing both ectoin and cyclodextrin, the denaturation rate was within 5%, indicating that the storage stability of EGF was improved.

As described above, the present invention has been described with specific details and limited examples and comparative examples, but these are provided only to facilitate a more general understanding of the present invention, and the present invention is not limited to the above examples. Those skilled in the art can make various modifications and variations from this description.

Accordingly, the spirit of the present invention should not be limited to the described embodiments, and the scope of the patent claims described below as well as all modifications that are equivalent or equivalent to the scope of this patent claim shall fall within the scope of the spirit of the present invention. .

Claims (10)

Contains EGF, ectoine and cyclodextrin dispersions,
A cosmetic composition for stabilizing EGF, wherein the weight ratio of EGF and ectoin is 1:30 to 1:60. delete delete According to paragraph 1,
A cosmetic composition for stabilizing EGF, wherein the weight ratio of EGF and cyclodextrin is 1:1 to 1:20. According to paragraph 1,
A cosmetic composition for stabilizing EGF, wherein the cyclodextrin dispersion contains polyol as a solvent. According to clause 5,
A cosmetic composition for stabilizing EGF, wherein the weight ratio of polyol and water contained in the cosmetic composition for stabilizing EGF is 1:1 to 1:10. According to clause 5,
The polyol includes one or more selected from the group consisting of glycerin, polyethylene glycol, butylene glycol, pentylene glycol, isopentyldiol, hexanediol, methylpropanediol, sorbitol, erythritol, inositol, maltitol, and xylitol. A cosmetic composition for stabilizing EGF. According to paragraph 1,
The cosmetic composition for stabilizing EGF is,
Preparing an EGF aqueous solution;
Mixing 30 to 60 weight of ectoine aqueous solution with the EGF aqueous solution based on 1 weight of the EGF; and
Mixing a cyclodextrin dispersion with the EGF aqueous solution; A cosmetic composition for stabilizing EGF, prepared according to a method comprising. According to clause 8,
The step of mixing the cyclodextrin dispersion with the EGF aqueous solution includes solubilizing the cyclodextrin using a polyol solvent. A cosmetic composition for stabilizing EGF. According to any one of claims 1 and 4 to 9,
The cosmetic composition for stabilizing EGF is in the form of a lotion, skin, lotion, nutritional lotion, nutritional cream, massage cream, essence, pack, or microneedle patch.