KR101979682B1 - Composition for preventing or improving Epiermolysis Bullosa comprising Rhus Semialata M. Extracts - Google Patents

Abstract

The present invention relates to a composition for preventing or ameliorating epidermolysis bullosa comprising a Rhus semialata M. extract as an active ingredient. The composition comprising a Rhus semialata M. extract as an active ingredient of the present invention has an effect of protecting the dermal-epidermal junction, preventing epidermolysis bullosa, and protecting markers of epidermal stem cells, which are degraded by external stress. Accordingly, the present invention provides a cosmetic and a pharmaceutical composition for preventing or ameliorating epidermolysis bullosa, comprising the Rhus semialata M. extract.

Description

[0001] The present invention relates to a composition for preventing or ameliorating epidermolysis epidermolysis,

The present invention relates to a composition for preventing or ameliorating epidermolysis epidermolysis caused by Rhus Semialata M. extract as an active ingredient, and more particularly, to a composition for promoting the production of extracellular matrix of dermal basement membrane, The present invention relates to a composition for preventing or ameliorating epidermal erythema epidermolysis, which has an effect of protecting stem cells and protecting the epidermal-dermal boundary.

Skin is the primary protective membrane of the human body and protects the internal organs of the body from changes in temperature and humidity, stimulation of the external environment such as ultraviolet rays and pollutants, and plays an important role in the maintenance of body homeostasis such as body temperature control. However, excessive physical and chemical stimuli, ultraviolet rays, stress, and nutritional deficiencies, which are externally received, degrade skin normal functions and promote skin aging such as loss of elasticity, keratinization and wrinkle formation. Especially, Is severely damaged.

The dermis-epidermal junction (DEJ) is the junction of the dermal and epidermal layers of the skin, which not only functions as a skin barrier but also affects the adhesion of the epidermis and dermis and the alignment of epidermal cells for re- Help. The basal keratinocyte located at the outermost part of the dermis-epidermal boundary is composed of fibronectin, nidogen, laminin / integrin, and the like by the anchoring filament of hemidesmosomes. It is connected to a transparent plate (Lamina lucida) composed of subunits. The components of the transparent plate are combined with a base plate (Lamina densa) which is a rigid collagen layer composed of collagen IV and collagen V again. The dermis binds to the dermal-epidermal boundary through collagen VII, a fixed fiber.

The understanding of the adherent complexes of the dermal-epidermal boundary, which is necessary for stable binding of epidermis and dermis, is the most important factor to understand epidermolysis. It is important that the presence of hemidesmosomes of basal keratinocytes in the epidermal layer for stable binding to the transparent plate, and the expression of fixed endothelial fibers on the basement membrane dermis side necessary for binding to the basal lamina. The absence of components of the adherent complex at the epidermal-dermal boundary or the factors that interfere with protein-protein interactions in the complex can significantly reduce the function of the skin barrier, resulting in minor skin trauma or skin damage due to exposure to contaminants do.

Stem cells are involved in overall skin health, including skin regeneration, skin damage recovery, wound healing, as well as hair color and regeneration. Human epidermal stem cells, which reside in the basement membrane of the skin, are used for cell proliferation through self-renewal and differentiation for the replacement of cells after skin damage or aging ), It plays an important role in maintaining the skin homeostasis of the epidermal layer. Although epidermal stem cells provide new cells to repair damaged tissues in response to tissue damage, some stem cells remain quiescent, protected from differentiation and senescence to prolong survival. However, humans have a limited number of stem cells. As the aging process progresses, the number of stem cells decreases and the number of internal aging factors causing UV irradiation, microorganism infection, environmental pollution, And their activities are also reduced by external threats. In particular, recent rapid destruction of the ozone layer increases the level of ultraviolet land reach. Although epidermal stem cells are highly resistant to DNA damage, persistent and repeated exposure of UVB induces damage to epidermal stem cells as well as extracellular matrix components that constitute the basement membrane.

Stem cell niche regulates stem cell proliferation and differentiation into lineage by inhibiting apoptosis or providing signal for stem cell regeneration to maintain the number of stem cells. . The stem cell is a special microenvironment composed of the right cell and the extracellular matrix proteins. In this microenvironment, the stem cell is a surface marker including keratin or integrin. By interacting with markers that are expressed inside the cell, they can communicate with their specific components by being stimulated to protect the stem cells from the danger. Stem cells can be characterized by stem cell specific marker gene expression and molecular properties. Integrin β1 and integrin α6, which are stem cell markers expressed in epidermal stem cells, adhere to the extracellular matrix of the basement membrane and are resistant to apoptosis by genotoxic stimuli, thus helping to survive for a long time. If the stem cells are separated from the basement membrane, they migrate away from the resting state and migrate to the skin surface. Several studies have shown that restoration of extracellular quality plays a very important role in the maintenance of stem cell function in the skin.

Epiermolysis Bullosa is a major hereditary connective tissue disorder that affects about 20 people per million in the United States. Normal epidermis and dermis are immobilized independently of each other by anchoring protein. In the case of patients, the lack of these anchoring proteins responsible for adherence of epidermis-dermis, Or external pressure, the blister is formed with severe pain corresponding to the third degree image. In addition, patients with epidermal blisters, epidermolysis, are at increased risk of skin cancer due to complications from chronic skin damage. The disease is divided into Epidermolysis Bullosa simplex, Junction Epidermolysis Bullosa, and Dystrophic Epidermolysis Bullosa. Simplified waterborne epidermolysis is the most common form of autosomal dominant form of keratin 5 and keratin 14, with blisters forming mainly in the hands and feet. It is a hereditary disease affecting laminin and collagen, characterized by formation of blisters in the basement lamina lucida, and is an autosomal thermoformed hereditary disease that occurs in infants and adults, If there is blisters on the road, the respiratory tract may also cause a risk of closure. In the case of epidermolysis bullosa, which accounts for about 30% of the disease, it is caused by a mutation of the collagen 7A1 (COL7A1) gene that expresses collagen 7, which is the main component of the anchoring fibril that forms a stable structure. (Pediatr. Int. 55, 234-237).

On the other hand, Rhus Semialata M. is a deciduous arboreous tree belonging to Rhus verniciflua. It is mainly distributed in Northeast Asia such as Korea, China and Japan. It is also used as a fountain for the production of salted salty taste and tofu in October. It is also called as umbrella, and the aphid is parasitized on the wings of the tree petiole to form a house of insects. It is also called a tree. Korean Patent Registration No. 10-1617152 discloses a cosmetic composition containing an extract of rhusi mushroom as an active ingredient in relation to the efficacy of fruit, leaves and branches of Rhus verniciflua. Korean Patent No. 10-0997641 discloses a cosmetic composition comprising a molecular encapsulated rusk extract A cosmetic composition containing an active ingredient was disclosed as a fermented product. A functional cosmetic ingredient extraction method was disclosed in Korean Patent No. 10-0872454. The contents of the activity of the dendrimer tincture compounds separated from rhusin 10-0873222 discloses a composition for preventing, ameliorating and treating arteriosclerosis, cancer and oxidative stress containing a dementia reintertifene compound, Korean Patent Registration No. 10-0685516 discloses a novel anticancer active triterpene compound and its The manufacturing method has been described.

However, there has been no report on epidermal stem cell protection and epidermal detachment prevention effect through promotion of extracellular matrix formation of rhizome basement membrane.

Accordingly, the inventors of the present invention confirmed that the extract of rushes increased the production of collagen IV, collagen VII and laminin of basement membrane and increased the expression of surface markers of epidermal keratinocytes. Thus, The present invention can be used as a composition for preventing epidermal blistering and epidermolysis due to reduction of collagen VII and collagen VII.

Accordingly, a technical problem to be solved by the present invention is to provide a safe substance without skin irritation, while being able to prevent or treat epidermal eruptive epidermolysis with an effect of protecting the epidermis-dermis boundary.

In order to solve the above-mentioned technical problem, the present invention provides a method for producing Rhus Semialata M. The present invention provides a composition for preventing or ameliorating epidermal eruptive epidermolysis, which comprises, as an active ingredient,

Preferably, the prevention or amelioration of epidermolytic epidermolysis is characterized by protecting the epidermis-dermis boundary, and protecting the epidermis-epidermis boundary means promoting the production of epidermal stem cells.

Preferably, the composition comprising the rhusi extract in the present invention may be a cosmetic composition or a pharmaceutical composition.

In the present invention, the " rattan extract " is an extract obtained by extracting rhusin according to a conventional method, and includes not only the extract extracted from rhusin but also its dry powder or any form thereof formulated therewith.

The Rhus verniciflua extract can be extracted with water or an organic solvent, and the extracted liquid can be used in a liquid form or can be used by concentration and / or drying. The organic solvent may be selected from the group consisting of methanol, ethanol, isopropanol, butanol, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, dichloromethane, N, N-dimethylformamide (DMF), dimethylsulfoxide , 3-butylene glycol, propylene glycol, or a mixed solvent thereof. The extract can be extracted at room temperature or with heating under the condition that the active ingredient of the extract is not destroyed or minimized.

The extraction method is not limited, and examples thereof include cold extraction, ultrasonic extraction, and reflux cooling extraction.

In the present invention, any part of the rhus major, leaves, roots, seeds and stems may be used.

According to a preferred embodiment of the present invention, the rusks extract is an ethanol extract obtained by adding 10 to 20 times volume of ethanol to rhodiola leaves and branches and extracting at room temperature for 20 to 30 hours.

The rumen extract of the present invention may be any extract, fraction, purified product, diluted solution, concentrated solution, or dried product obtained in each step of extraction, fractionation, or purification (separation, fractionation).

The composition of the present invention may contain 0.0001 to 20% by weight, more preferably 0.001 to 10% by weight, and most preferably 0.1 to 5% by weight, based on the total weight of the rhusi extract. When the extract is less than 0.0001% by weight of the total composition, the effect of producing DEJ is too weak. When it exceeds 20% by weight, the increase in the effect of the extract is insufficient, There is a problem. More preferably, the rhusi extract is contained in an amount of 0.001 to 5% by weight based on the whole composition.

According to a specific embodiment of the present invention, the rhusi extract has an effect of promoting epidermal-dermal boundary formation, and has an effect of protecting the markers of epidermal stem cells, which are degraded by epidermal blister-like epidermolysis prevention and external stress.

According to one preferred embodiment of the present invention, the present invention relates to a cosmetic composition for improving DEJ comprising rhusi extract as an active ingredient, wherein the skin skin stem cell protection is selected from the group consisting of collagen IV, collagen VII and laminin Which is characterized in that it is prevented or treated by the stimulation of production.

In the present invention, the above-mentioned " active ingredient " means a component that exhibits the desired activity alone or can exhibit activity together with a carrier which is itself inactive.

According to one embodiment of the present invention, there is provided a cosmetic composition comprising rumba extract as an active ingredient.

The cosmetic composition according to one embodiment of the present invention may contain, in addition to the rusks extract as an active ingredient, conventional additives such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavors, May be further added.

The cosmetic composition of the present invention can be prepared into any of the formulations conventionally produced in the art and can be used as a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, , Oil, powder foundation, emulsion foundation, wax foundation and spray, but is not limited thereto. More specifically, it can be prepared as a nutritional cream, a convergent lotion, a soft lotion, a lotion, an essence, a nutritional gel or a massage cream.

When the formulation of the present invention is a paste, cream or gel, use is made of an animal oil, vegetable oil, wax, paraffin, starch, tragacanth gum, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide .

According to one embodiment of the present invention, the cosmetic may be a hair tonic, a hair nutrition lotion, a scalp treatment, a hair treatment, a hair rinse, a hair shampoo or a hair lotion.

When the formulation of the present invention is a powder or a spray, tosse, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In the case of a spray, in particular, chlorofluorohydrocarbons, propane / Propane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, a solubilizing agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.

In the case where the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Castellulose, aluminum metahydroxide, bentonite, agar or tracert, etc. may be used.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters.

As one specific use of the present invention, the composition of the present invention can be used as a pharmaceutical composition for scalp protection.

When the composition of the present invention is used as a pharmaceutical composition, a pharmacologically acceptable carrier other than the rusks extract may be further included as an active ingredient. The pharmaceutically acceptable carriers to be contained in the pharmaceutical composition of the present invention are those conventionally used in the present invention and include carbohydrate-type compounds (e.g., lactose, amylose, dextrose, sucrose, sorbitol, mannitol, starch, Corn oil, cottonseed oil, cottonseed oil, cottonseed oil, cottonseed oil, cottonseed oil, cottonseed oil, cottonseed oil, cottonseed oil, cottonseed oil, cottonseed oil, But are not limited to, polyethylene glycol, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like. The pharmaceutical composition of the present invention may further contain a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc., in addition to the above components. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington ' s Pharmaceutical Sciences (19th ed., 1995).

A preferred dosage of the pharmaceutical composition of the present invention is within the range of 0.001 to 100 mg / kg on an adult basis. When the composition is an external preparation, it is preferably applied in an amount of 1.0 to 3.0 ml on an adult basis once to five times a day, and continued for 1 month or more. The dose does not limit the scope of the present invention.

The formulation of the pharmaceutical composition of the present invention may be in the form of powders, granules, tablets, capsules, emulsions, oral preparations such as syrups and aerosols, external preparations such as ointments and creams, sterilized injection solutions, And may be formulated and used in any form suitable for the preparation.

On the other hand, the ruka extract of the present invention has no toxicity and can be safely used for long-term use, so that it can be safely used in cosmetics or pharmaceuticals.

As described above, the rhusi extract of the present invention has an effect of protecting the epidermis-dermis boundary, and has an effect of protecting the marker of epidermal stem cells, which is degraded by external stress, in preventing epidermal blister-like epidermolysis. Accordingly, the present invention provides a cosmetic composition and a pharmaceutical composition for preventing or ameliorating epidermal eruptive epidermolysis caused by Rhodiola extract.

FIG. 1 is a graph showing the results of comparing the amount of laminin and collagen produced in epidermal stem cells after treatment with rattan extract, and the effect of stimulating laminin production of fibroblasts reduced by ultraviolet irradiation according to pretreatment of rattan extract.
FIG. 2 is a graph showing the promoting effect of epidermal stem cell marker expression and the stimulation of extracellular matrix synthesis in the basement membrane of rattan extracts tested in the human skin epidermis.
Fig. 3 is a diagram showing the protective effect of rusks extract against the expression of surface markers reduced by ultraviolet irradiation, confirming the surface marker expression pattern as a characteristic of epidermal stem cells.

Hereinafter, embodiments of the present invention will be described in detail to facilitate understanding of the present invention. However, the embodiments according to the present invention can be modified into various other forms, and the scope of the present invention should not be construed as being limited to the following embodiments. Embodiments of the invention are provided to more fully describe the present invention to those skilled in the art.

Example 1: Preparation of rattan extract

1kg of leaves and branches of Rhus javanica (Jeju Island) were washed, dried and pulverized and powdered. 10L of 70% ethanol was added and the mixture was stirred at room temperature for 24 hours. The extract solution was filtered through a filter paper (Whatman, Advantes, No. 2), concentrated under reduced pressure, and lyophilized to prepare a rattan extract.

<Experimental Example 1> Promoting action of collagen IV, collagen VII and laminin in the dermal fibroblast of Rhus verniciflua extract in vitro ) Measure

Culture supernatant of epidermal keratinocytes treated with rattan extract for 24 hours in a 96-well plate coated with antibody against collagen IV, collagen VII and laminin and culture supernatant irradiated with UVB were dispensed and incubated at 37 ° C for 1 hour and incubated to attach the respective extracellular matrix molecules, followed by an experiment as directed in a commercial ELISA kit (Cloud-Clone Corp., USA). After substrate was added, color was developed and absorbance was measured at 450 nm. All experiments were repeated twice. The results are shown in Fig.

FIG. 1 is a graph showing the results of comparing the amount of laminin and collagen produced in epidermal stem cells after treatment with rattan extract, and the effect of stimulating laminin production of fibroblasts reduced by ultraviolet irradiation according to pretreatment of rattan extract. As shown here, the expression of laminin and collagen IV was increased in the epidermal stem cells treated with rattan extract, and the laminin produced from the dermal fibroblasts reduced by UVB irradiation was increased by pretreatment of rattan extract. These results suggest that Rhus japonica extract not only increases the expression of integrin molecules in epidermal stem cells but also increases the expression of extracellular matrix of basement membrane.

&Lt; Test Example 2 > Ex-vivo measurement of expression of p63, integrin beta1 and extracellular matrix in the skin excretory part of rattan extract

Human skin epidermis was obtained from a 54 - year - old Caucasian female who underwent abdominal implantation of 11 mm (± 1 mm) and cultured in BEM culture medium at 37 ° C and 5% CO ₂ . Rhus verniciflua extract was applied to the surface of the skin of the skin and repeatedly treated at 0, 2, 5, 7 and 9 days. The cells were collected on day 9 and fixed in formalin solution. The sections were cut to a thickness of 7 μm and stained with p63, integrin β1, collagen IV, collagen VII and laminin, respectively, and observed with fluorescence microscope.

FIG. 2 is a graph showing the promoting effect of epidermal stem cell marker expression and the stimulation of extracellular matrix synthesis in the basement membrane of rattan extracts tested in the human skin epidermis. As shown here, the expression of p63 and integrin β1, which plays an important role in the differentiation and migration of keratinocytes, was increased by treatment with rattan extract. In addition, it was confirmed that the production of extracellular matrix such as laminin, collagen IV and collagen VII was increased in the basement membrane by treatment of rattan extract. As a result, the extracts of rusks showed very increased production of laminin and collagen VII and a slight increase in collagen IV and increased the production of p63 and integrin β1 protein between the basement membrane and basal layer of the dermal - epidermal boundary.

<Test Example 3> Expression Patterns of Surface Marker Cell Surface of Epidermal Stem Cells of Rhus verniciflua extract in vitro ) Measure

Expression patterns of integrin β1, α2, α6, cytokeratin 15 and p63, which are known as surface markers, were examined to verify the characteristics of epidermal stem cells.

Fig. 3 is a diagram showing the protective effect of rusks extract against the expression of surface markers reduced by ultraviolet irradiation, confirming the surface marker expression pattern as a characteristic of epidermal stem cells. As shown here, each of the surface markers was significantly expressed in the early human epidermal keratinocytes, and the expression of integrin β1 and α2 was decreased by UVB irradiation. In order to test the effect of the extract on the integrin expression of rhus chinensis extract, the rhusi extract was pretreated for 24 hours before UVB irradiation, and the expression patterns of integrin β1 and α2 were then fluorescently stained with a specific antibody, (Flow cytometry, Becton Dickinson, USA) and analyzed with CellQuestTM Pro software. The extracts of rhus chinensis showed that the expression of integrin molecules decreased by UVB irradiation in a concentration dependent manner. It was confirmed that the extract of Rhus verniciflua was obtained by up-regulating the surface markers of epidermal stem cells.

Formulation Example 1 Cosmetic preparation

1-1. Softening longevity

ingredient weight% Rhodiola Leaf Branch Extract 0.05 glycerin 3.0 Butylene glycol 2.0 Propylene glycol 2.0 Carboxyvinyl polymer 0.1 ethanol 10.0 Triethanolamine 0.1 Preservatives, micronutrients, trace fragrances and trace water 82.75 sum 100.0

1-2. Nutritional lotion

ingredient weight% Rhodiola Leaf Branch Extract 0.05 Wax 4.0 Polysorbate 60 1.5 Sorbitan sesquioleate 0.5 Liquid paraffin 5.0 Squalane 5.0 Caprylic / capric triglyceride 5.0 glycerin 3.0 Butylene glycol 3.0 Propylene glycol 3.0 Carboxyvinyl polymer 0.1 Triethanolamine 0.2 Preservatives, micronutrients, trace fragrances and trace water 69.65 sum 100.0

1-3. Nourishing cream

ingredient weight% Rhodiola Leaf Branch Extract 0.05 Wax 10.0 Polysorbate 60 1.5 Sorbitan sesquioleate 0.5 Liquid paraffin 10.0 Squalane 5.0 Caprylic / capric triglyceride 5.0 glycerin 5.0 Butylene glycol 3.0 Propylene glycol 3.0 Triethanolamine 0.2 Preservatives, micronutrients, trace fragrances and trace water 56.75 sum 100.0

1-4. Massage cream

ingredient weight% Rhodiola Leaf Branch Extract 0.05 Wax 10.0 Polysorbate 60 1.5 Sorbitan sesquioleate 0.8 Liquid paraffin 40.0 Squalane 5.0 Caprylic / capric triglyceride 4.0 glycerin 5.0 Butylene glycol 3.0 Propylene glycol 3.0 Triethanolamine 0.2 Preservatives, micronutrients, trace fragrances and trace water 27.45 sum 100.0

1-5. pack

ingredient weight% Rhodiola Leaf Branch Extract 0.05 Polyvinyl alcohol 13.0 Sodium carboxymethylcellulose 0.2 Allantoin 0.1 ethanol 5.0 Nonyl phenyl ether 0.3 Preservatives, micronutrients, trace fragrances and trace water 81.35 sum 100.0

Formulation Example 2: Pharmaceutical preparation

2-1. Manufacture of Powder

ingredient Content (g) Rhodiola Leaf Branch Extract 2 Lactose One

The above components were mixed and packed in airtight bags to prepare powders.

2-2. Manufacture of tablets

ingredient Content (mg) Rhodiola Leaf Branch Extract 100 Corn starch 100 Lactose 100 Magnesium stearate 2

After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.

2-3. Preparation of capsules

ingredient Content (mg) Rhodiola Leaf Branch Extract 100 Corn starch 100 Lactose 100 Magnesium stearate 2

After mixing the above components, the capsules were filled in gelatin capsules according to the conventional preparation method of capsules.

Claims (4)

Rhus Semialata M. The present invention relates to a cosmetic composition for preventing or ameliorating epidermal eruptive epidermal dissipation. The method according to claim 1,
Characterized in that the prevention or improvement of the epidermis-induced epidermal dissipation is to protect the epidermis-dermis boundary. Rhus Semialata M. Extracts) as an active ingredient for the prevention or treatment of epidermal erythema epidermolysis. The method of claim 3,
A pharmaceutical composition for preventing or treating epidermal dermatomyositis, which is characterized in that the prevention or treatment of epidermal hydropsic epidermolysis is to protect epidermis-dermis boundary.

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