KR102272237B1 - Composition for improved atopy skin and skin moisturizing comprising natural extract - Google Patents

Abstract

The present invention relates to a composition for improving atopic skin and moisturizing the skin comprising a natural extract, wherein the ability to inhibit the release of histamine and β-hexosaminidase by mast cells or neutrophils by using calcined coral powder, Bupyeongcho and aloesincense. As shown, it is possible to improve atopic skin, and it is possible to provide a composition for improving atopic skin and moisturizing skin that strengthens the skin barrier and increases moisture content.

Description

Composition for improving atopic skin and moisturizing skin containing natural products {COMPOSITION FOR IMPROVED ATOPY SKIN AND SKIN MOISTURIZING COMPRISING NATURAL EXTRACT}

The present invention relates to a composition for improving atopic skin and moisturizing the skin containing a natural product. In more detail, it is possible to improve atopic skin by using calcined coral powder, Bupyeongcho, and amethyst to inhibit the release of histamine and β-hexosaminidase of mast cells or neutrophils, and improve atopic skin by increasing the moisture content. And it relates to a composition for moisturizing the skin.

The skin of the human body is an organ that functions as a protective film to protect the living body from the external environment, and prevents the loss of biological components such as water and electrolytes in the human body, and at the same time prevents the intrusion of harmful substances from the outside. The structure of the skin is largely divided into the epidermis, cortex, and subcutaneous fat. The epidermal layer is composed of keratinocytes and melanocytes, and the keratinocyte layer, which is the outermost layer of the skin, is in direct contact with the external environment and acts as a primary barrier against penetration of physical, chemical, or substances. In addition, it is also required to maintain flexibility by preventing moisture loss to the outside of the skin and retaining moisture by forming intercellular lipids into a lamellar layer on their own. As the importance of the stratum corneum is emphasized, research on it is being actively conducted. In particular, studies on the structure and function of lipids between keratinocytes existing in the stratum corneum are actively being conducted. The keratinocyte intercellular lipid exhibits a multi-layered lipid film form called a lamellar structure, and in particular, as ceramide is known to play an important role in the function of keratinocyte intercellular lipid, various moisturizers using this have been developed.

On the other hand, in order to understand the mechanism of atopic dermatitis, the opinion that it is caused by an immunological abnormality, particularly inflammation involving a Th2 immune response, has prevailed. It is suggested that it is caused by an abnormality in the bast skin barrier. That is, in the skin of atopic dermatitis patients, a decrease in the moisture content of the epidermis and a malfunction of the skin barrier are induced by genetic abnormalities and lipid abnormalities such as ceramides constituting the skin barrier. Due to this, the skin penetration of antigens increases, the immune response increases, and various aggravating factors such as changes in skin pH caused by the use of detergents aggravate the deterioration of the skin barrier.

Atopic dermatitis is an inflammatory disease accompanied by pruritus, dry skin, eczema, etc., or a disease caused by abnormalities in the stratum corneum, the outermost protective barrier of the skin, due to genetic, environmental, and immunological causes. In climate, it tends to be more severe. Such atopic dermatitis is caused by a complex entanglement of various causes, and remission and recurrence are repeated. Allergic diseases caused by atopic predisposition include allergic dermatitis, allergic rhinitis, asthma, allergic conjunctivitis, and atopic urticaria, and these diseases may appear alone or simultaneously with other diseases. Atopic dermatitis affects a very large number of people, with 0.5 to 1% of the total population and 5 to 10% of children suffering from atopic dermatitis. 50% of patients heal within two stones, but 25% continue into adolescence, and the remaining 25% continue without atopic dermatitis even into adulthood.

The cause of such atopic dermatitis is not clearly known, but it has been found that there are mainly many genetic factors and it is related to a deficiency of the immune system. In addition, it seems to be caused by a complex action of dry skin, the characteristic of feeling itchy skin more easily than normal people, infection by bacteria, viruses, fungi, emotional factors, and environmental factors.

The main symptoms of atopic dermatitis are severe itching, dry skin, rash, oozing, scabs, and scaly skin (scaly). In particular, the severe itching of atopic dermatitis causes psychological damage and makes it difficult to live a normal life.

Since atopic constitution is fundamentally very difficult to treat, atopic dermatitis is being controlled through appropriate treatment while avoiding triggers rather than aiming for a cure. Common atopic dermatitis synergists currently used include moisturizing agents, antihistamines that reduce pruritus (itchiness), topical steroids that have therapeutic effects through anti-inflammatory, vasoconstriction, and immunosuppressive actions. Drug therapy such as steroids, antihistamines, and antibiotics is mainly used. Steroid drugs such as glucocorticoids and cyclospoline can cause diabetes and high blood pressure, and cause serious side effects such as Cushing's syndrome, ophthalmic diseases (cataracts, glaucoma), kidney and liver toxicity. In particular, steroids (adrenal corticosteroids) have anti-inflammatory and immunosuppressive effects and have excellent effects, but side effects such as skin weakness, systemic hormonal symptoms, and addiction may appear when applied for a long period of time. Antihistamines relieve itching symptoms by preventing histamine from being released from mast cells, but are used as a temporary measure and may have side effects such as insomnia, anxiety, and loss of appetite when taken for a long time.

Accordingly, there is a demand for a new concept atopic dermatitis treatment that is effective for atopic dermatitis and has no side effects.

KR 10-2036826 B1

It is an object of the present invention to provide a composition for improving atopic skin and moisturizing the skin containing a natural product.

Another object of the present invention is to improve atopic skin by exhibiting histamine release and β-hexosaminidase release inhibitory ability of mast cells or neutrophils by using calcined coral powder, Bupyeongcho and amethyst, and increase water content It is to provide a composition for improving atopic skin and moisturizing the skin.

In order to achieve the above object, the composition for improving atopic skin and moisturizing the skin according to an embodiment of the present invention contains a natural extract selected from the group consisting of calcined coral powder, Bupyeongcho, amethyst and mixtures thereof as an active ingredient. .

The atopic skin is a symptom selected from the group consisting of dry skin, pruritus, dermatophytosis, and eczematous lesions.

The composition exhibits an alleviating effect of atopic skin through inhibition of histamine and β-hexosaminidase.

The extract is extracted using an extraction solvent selected from the group consisting of water, C ₁ to C _{6 lower alcohols, and mixtures thereof.}

A cosmetic composition according to another embodiment of the present invention is prepared by using the composition for improving atopic skin and moisturizing the skin.

A moisturizing cream according to another embodiment of the present invention is prepared using the composition.

A skin regeneration cream according to another embodiment of the present invention is prepared using the composition.

Hereinafter, the present invention will be described in more detail.

As used herein, the term 'extract' has the meaning commonly used as a crude extract in the art as described above, but in a broad sense also includes a fraction obtained by additionally fractionating the extract. That is, the plant extract includes not only those obtained using the above-described extraction solvent, but also those obtained by additionally applying a purification process thereto. For example, a fraction obtained by passing the extract through an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatography (prepared for separation according to size, charge, hydrophobicity or affinity), etc. Fractions obtained through various purification methods are also included in the plant extract of the present invention.

The extract used in the present invention may be prepared in a powder state by an additional process such as distillation under reduced pressure and freeze-drying or spray-drying.

As used herein, the term "inflammatory disease" refers to a pathology caused by an inflammatory response specified as a local or systemic biological defense response to an external physical or chemical stimulus or infection or autoimmunity from external infectious agents such as bacteria, fungi, viruses, and various allergens. It can be defined as a symptom. This inflammatory response involves activation of various inflammatory mediators and immune cell-related enzymes (iNOS, COX-2), secretion of inflammatory mediators (secretion of NO, TNF-α, IL-6, etc.), fluid infiltration, cell migration, and tissue It is accompanied by a series of complex physiological reactions such as destruction, and is externally manifested by symptoms such as erythema, pain, edema, fever, and deterioration or loss of specific functions of the body.

In order to achieve the above object, the composition for improving atopic skin and moisturizing the skin according to an embodiment of the present invention contains a natural extract selected from the group consisting of calcined coral powder, Bupyeongcho, amethyst and mixtures thereof as an active ingredient. .

The calcined coral powder is a water-soluble calcium oxide (CaO) and a small amount of inorganic minerals (about 70 kinds) that are soluble in water by calcining weathered sedimentary coral limestone formed by coral reefs and coral sands that have only grown in clean waters since the geological era hundreds of millions of years ago. It is a clean natural mineral that contains this mineral, and it is a comprehensive mineral that makes the natural environment such as people, animals, and plants a healthy and pollution-free environment.

Bupyeongcho (Spirodela polyrhiza) is an annual floating aquatic plant belonging to the Lepidoptera family. The plant has an obovate shape as wide as a leaf. It grows in groups of 2-6 in rice paddies or ponds with a unique breeding method and grows wild in Korea. . Also, it is a small grass floating on the water surface. The outpost of frogfish is also called Bupyeongcho. Physiologically active ingredients of Bupyeongcho include flavonoid, anthocyanin and tannin in the forearm of frogfish, campesterol of sterol, β-sitosterol, apigenin-7-O-ß-D-glucoside of sterol, apigenin-7-O-ß-D-glucoside of flavones-O-glycoside, and luteolin-7- It has been reported that O-β-D-glucoside and the like are contained. In addition, components such as chlorophyll, potassium acetate and potassium chloride are also known, which have been reported to have immune and anticancer effects as well as cardiotropic and haemolytic effects as an oriental medicine action. Recently, it has been reported that the antioxidant effect of lotus leaf extract, a perennial aquatic plant of the water lily family, and the anti-aging effect of the extract and its fractions of the lotus leaf extract, which is an annual herb from the fruit of the water lily, and its fractions. as well as for the treatment of atopic skin

Aquilariae Agallochae Lignum Resinatum is a trunk tree in which the resin of aloes tree has been deposited, and the resin naturally secreted from the aloes tree is deposited to form a histologically hard mass in the heartwood of aloes tree. Since the old days, kings and royals have smoked aloes before the onset of illness and taken an appropriate amount to drive away bad energy and prevent clogging of the acupuncture points. It has excellent effects compared to the amount and has excellent therapeutic and preventive effects, but it is rare. Because of this, it was reserved for the upper class. In addition, when the qi is heo, circulation is not smooth, or when diseases such as asthma, ascites, constipation, thyroid disease, kidney disease, or lack of yang are caused by musical instruments, achimhyang alone has a significant effect. In particular, it is known to exert an excellent effect on patients undergoing renal dialysis due to renal failure or ascites due to cirrhosis of the liver.

In the case of using the natural extract, it is possible to improve atopic skin by showing an anti-inflammatory effect by the active ingredient contained in the natural extract, and at the same time enhances the skin barrier and increases the moisture content, and at the same time has excellent human stability.

Preferably, the composition of the present invention may include 100 parts by weight of the calcined coral powder extract, 100 parts by weight of the Bupyeongcho extract and 100 parts by weight of the aloes root extract.

Inflammation refers to a tissue response to damage when a foreign antigen such as bacteria or virus enters the body from the outside or when cells or tissues are damaged by any cause. Inflammatory cells release various proinflammatory mediators such as reactive nitrogen and oxygen species that cause DNA damage in tumor initiation and promotion. Inflammation is largely divided into acute inflammatory and chronic inflammatory. When a chronic inflammatory response occurs, proinflammatory cytokines and chemokines are excessively released, causing damage to surrounding tissues and invasion, metastasis, and migration of tumor cells. It also promotes angiogenesis and aids in tumor growth.

In general, in the inflammatory response, lipopolysaccharide (LPS) is an endotoxin derived from the cell wall of Gram-negative bacteria and activates the transcription factor NF-κB by binding to toll-like receptor 4 (TLR4), iNOS and COX-2 It induces the expression of NO, inflammatory cytokines, and various inflammatory regulators such as PGE₂.

Moreover, NO and superoxide radicals generated from endothelial cells, neutrophils and macrophages during the inflammatory process generate peroxynitrite (ONOO-), a highly reactive oxidizing agent, to cause pathophysiological tissue. Therefore, inducible nitric oxide synthase (iNOS) and cyclooxygease-2 (COX-2) genes are considered as representative inflammatory factors, and substances that inhibit their expression are highly evaluated as potential useful anti-inflammatory agents.

The calcined coral powder, Bupyeongcho and aloes root extract concentration-dependently inhibit the production of NO, inflammatory (IL-1β, IL-6, TNF-α), PGE₂, COX-2 and iNOS, and can be used as an anti-inflammatory agent. .

The PGE₂ is one of the inflammatory mediators and is synthesized by the enzymatic action of phospholipase A2. It starts with the production of arachidonic acid from membrane phospholipids.

The arachidonic acid becomes prostaglandin G2 by enzymatic action and again becomes prostaglandin H2, an unstable metabolite.

These two processes are catalyzed by cyclooxygenase (COX), and there are two or more isoenzymes of COX. Among them, COX-1 is continuously expressed to regulate platelet aggregation, gastric mucosa, and renal function. It is responsible for the physiological functions of the back, and COX-2 is expressed by stimuli such as inflammation.

PGH₂ produced by COX is an unstable intermediate metabolite and is metabolized into PGE₂, PGD₂, PGI₂, PGF₂, thromboxane A2 (TXA₂), etc. by various prostanoid synthases depending on the type of cell or stimulation.

In addition, three types of isoforms of PGE synthase (PGES) involved in the metabolism of PGH₂ to PGE₂, cytosolic PGES (cPGES), microsomal PGES-1 (mPGES-1), and mPGES-2, are known. Among mPGES-1, expression is increased by inflammatory stimuli such as lipopolysaccharid (LPS), inflammatory cytokines such as IL-1β and TNF-α, and nitric oxide (NO), and is functionally closely related to COX-2. It is closely linked to the production of PGE₂.

That is, the production of PGE₂ is functionally closely linked with COX-2 and plays an important role in the inflammatory response.

Accordingly, the present invention suppresses the production of COX-2 and iNOS, and suppresses the expression of inflammatory cytokines (Cytokine), nitric oxide (NO) and COX-2 by using calcined coral powder, Bupyeongcho, and aloesae as active ingredients. As it is inhibited, an excellent anti-inflammatory effect may be exhibited.

The atopic skin is a symptom selected from the group consisting of dry skin, pruritus, dermatophytosis, and eczematous lesions.

Since atopic skin is chronically dry, eczema is easily induced by various stimuli, it has severe itching, and has the characteristics of repeated recurrence. These pruritus and eczema are repeatedly weakened by dry skin and, if not properly managed, eventually progress to subacute and chronic lesions such as lichenification, pigment change, and erythroderma. With immunological abnormalities, superficial folliculitis, phytilosporum folliculitis, epilepsy, superficial dermatophytosis, warts, and recurrent herpes simplex are common. Bacterial infection is usually caused by coagulase-positive Staphylococcus aureus, and it mainly appears as folliculitis lesions with superficial pruritus on the arms and legs, and is rarely accompanied by deep tissue abscess and cellulitis, but is accompanied by severe recurrent annular eczema lesions. In some cases, it may cause recurrent cellulitis with systemic symptoms.

In addition, atopic dermatitis is an inflammatory disease accompanied by pruritus, dry skin, eczema, etc., or a disease caused by abnormalities in the stratum corneum, the outermost skin barrier, due to genetic, environmental, or immunological causes. tends to be more severe in dry climates. Therefore, the use of a moisturizer to improve the dryness of the skin of atopic dermatitis and the increase in the recovery of the skin barrier function occupies an important position in the treatment of atopic dermatitis.

The dry skin refers to a skin condition in which discomfort can be felt due to dryness, and refers to a condition in which moisture in the skin is insufficient to 10% or less of normal. Clinically, it refers to a skin condition with small red spots and fissures, with scales and a rough surface. If you have a skin disease or systemic disease that causes dry skin, it is important to treat the underlying disease. The basic principle of general dry skin treatment is to supply and maintain moisture in the stratum corneum while minimizing the loss of moisture in the stratum corneum. It is helpful to supply moisture directly to the skin through a shower or bath, but if there is no ability to keep the supplied moisture, showering or bathing is not helpful. Rather, it washes away the natural moisturizing factors, lipids of the stratum corneum, sebum, etc. may become drier. Therefore, in order to maintain the moisture and lipid components of the skin, it is better not to forcefully remove the stratum corneum. In addition, it is important to reduce excessive washing and soap use, protect the skin from the external harmful environment, and use an appropriate moisturizer. If dry skin is not controlled, it can progress to dermatitis.

Skin pruritus, also called skin itch, refers to a chronic skin disease in which skin itch is the main symptom without primary dermatitis, and is accompanied by secondary scratches and scabs. Wind and heat loss or damp heat loss is concentrated in the base, or wind and wind stays in the body for a long time and turns into fire, drying the essence and blood. It is caused by not being able to nourish the donation. It appears as paroxysmal, and worsens when exposed to heat or consuming stimulant foods or alcohol. It can lead to secondary pigmentation, lichen-like changes, and eczema-like changes as it ages. Systemic symptoms may include sleep disturbances, nervous breakdowns, and loss of appetite. Local symptoms include pruritus of the anus, pruritus of the scrotum or genitalia, and pruritus of the head, legs, and palms. It is often seen in adults, especially the elderly.

Dermatomycosis is a phenomenon in which erythematous redness and wheal (edema) occur in the affected area when a physical stimulus is applied to the skin, and it is a type of urticaria. When the skin is scratched or pressed with more than a certain amount of pressure, it is localized to the area and becomes itchy, red, and swollen similar to hives. As a result, it looks like writing on the skin. Even mild irritation can occur anywhere on the skin of the body, and if it is scratched because it is itchy, the symptoms of hives may appear even more severe. It occurs in about 5% of the Korean population and usually shows a chronic course.

The composition exhibits an alleviating effect of atopic skin through inhibition of histamine and β-hexosaminidase.

Atopic dermatitis is the most common chronic inflammatory skin disease, with a prevalence of about 20% in children and 3% in adults. Characteristic clinical symptoms include pruritus of the skin, lichenification of the skin, redness of the skin, and chronic skin bacterial infection. Recently, defects in the skin barrier are recognized as one of the most important features. The latest view on pathophysiology is that abnormal differentiation of skin epithelial cells causes a defect in the skin barrier, which leads to the penetration of allergens to induce the release of histamine and β-hexoamidase and an inflammatory response, and systemic immunological that the reaction takes place.

In particular, histamine, which is important in allergic and atopic diseases, is a type of active amine and is secreted from mast cells or basophils. Histamine exists as intracytoplasmic granules, and when an external stimulus (stress) is applied, it is secreted out of the cell, causing an allergic reaction or inflammation in addition to the reduction of blood pressure due to contraction of smooth muscle and vasodilation. In addition, it has been found that histamine secretion is stimulated by external stimuli such as sound or light or internal stimuli such as a feeling of hunger and an increase in body temperature. When histamine is secreted too much, it binds to a protein called the histamine first receptor (H1 receptor) and causes allergic symptoms such as runny nose, hives, itching, pain, and asthma due to bronchoconstriction. The initial symptoms of various dermatitis including atopic dermatitis are caused by histamine secreted from mast cells, and it is common to use antihistamine drugs for treatment.

Preferably, the composition is any one selected from the group consisting of Philadelphus schrenkii Rupr. var. schrenkii extract, Ulmus laciniata (Trautv.) mayr) extract, Callicarpa shiraswana Makino extract, and mixtures thereof. further includes

Philadelphus schrenkii Rupr. var. schrenkii is a deciduous shrub growing in valleys throughout Korea. It grows in a place with good drainage of soil, high ambient humidity, and abundant humus. It is about 2~4m tall, and the leaves are alternate phyllotaxis, 7~13cm long and 4~7cm wide. The surface is green with few hairs, and the back side is light green with fine hairs and is egg-shaped. Branches are divided into two, small branches are brown and hairy, and biennial branches are gray and peeled. The flowers are white and fragrant in white flowers on long stalks at the top or where the leaves are attached. The fruit is 0.6-0.9 cm long and 0.4-0.5 cm in diameter in September and hangs in an oval shape. It is mainly used for ornamental purposes, and the young leaves are used for food.

Ulmus laciniata(Trautv.) mayr) is a deciduous broad-leaved arboreous tree in the Elm family of the dicotyledonous bicotyledonous group, and grows in the valley below the hillside. The height is about 20 m and the diameter is about 1 m, and the small branches are light brown. The leaves are alternate, long oval or inverted egg-shaped, broad, with three deep dents on the edge and sharply pointed. Also, it is 10~20cm long, has sharp double serrated edges, has rough and hairy surface, and has a light green back. Flowers are bisexual and bloom from April to May, and inflorescences are divided into 5-6 pieces. 5-6 stamens are magenta, and the style is divided into two. The fruit is a citrus fruit, flat, 1.5cm long, egg-shaped, and ripens in May to June. It is used for tools and firewood, and the bark is used for medicinal purposes.

Callicarpa shiraswana Makino grows in the forest at the foot of the island in the southern coast of Korea. It has weak cold resistance but strong sound resistance, so it grows well on the beach. Leaves are opposite, egg-shaped, oval or oval-lanceolate, pointed pointed, round or acute, 3-12cm long and 2.5-5cm wide, with fine dotted spots on both sides, short hairs on the surface, dense hairs on the back side, and the edge There are sharp teeth on the leaf, and the petiole is 5~10mm in length, with dense hairs. Flowers are bloomed in August, and the inflorescence is axillary and densely clustered with virgin hairs, and the calyx is deeply divided into 5 parts, and the hairs of the female or idols are dense. The corolla is 4-5mm long, light purple, and the plate tube is almost the same length as the calyx, and the stamen is the same length as the corolla.

The composition for improving atopic skin and skin moisturizing is used in addition to calcined coral powder, Bupyeongcho extract, and aloes hyang extract, further comprising any one selected from the group consisting of gogwang tree extract, nanthu tree extract, serrata oleifera extract, and mixtures thereof. And, when used as a complex extract, it can improve atopic skin and improve skin moisturizing effect.

Preferably, the composition of the present invention is based on 100 parts by weight of calcined coral powder extract, 100 parts by weight of Bupyeongcho extract and 100 parts by weight of aloes root extract, 40 to 60 parts by weight of gogwang tree extract, 40 to 60 parts by weight of nanthu tree extract, It may be included in an amount of 40 to 60 parts by weight. When used as a complex extract within the above range, the atopic skin improvement and skin moisturizing effect are further increased due to the complex action between each component.

The extract is extracted using an extraction solvent selected from the group consisting of water, C ₁ to C _{6 lower alcohols, and mixtures thereof.}

Specifically, washing the natural product to prepare the natural extract; drying after washing; pulverizing the natural product after drying; leaching the pulverized product using an organic solvent; drying the sample after leaching; leaching with water; And including the step of leaching, it is possible to obtain a natural extract.

The natural extract extracted using the organic solvent may further include the step of performing fractionation using an organic solvent.

The method for preparing the extract may be a conventional extraction method in the art, such as an ultrasonic extraction method, a leaching method, and a reflux extraction method. Specifically, it may be an extract obtained by extracting a natural product from which foreign substances have been removed by washing and drying with water, an alcohol having 1 to 6 carbon atoms, or a mixed solvent thereof, and may be an extract extracted by sequentially applying the solvents to the sample.

The reflux extraction method is water, based on 100 mL of an alcohol having 1 to 6 carbon atoms, 10 to 30 g of a pulverized product of a natural product, a reflux time of 1 to 3 hours, and 50 to 100% alcohol or water having 1 to 6 carbon atoms. More specifically, based on 100 mL of an alcohol having 1 to 6 carbon atoms or 100 mL of water, 10 to 20 g of a pulverized product of a natural product, a reflux time of 1 to 2 hours, and 70 to 90% of an alcohol or water having 1 to 4 carbon atoms.

The leaching method is performed at 15 to 30° C. for 24 to 72 hours, and water or 50 to 100% alcohol having 1 to 6 carbon atoms is used as an extraction solvent. More specifically, the process is carried out at 20 to 25° C. for 30 to 54 hours, and the extraction solvent is water or 70 to 80% alcohol having 1 to 6 carbon atoms.

In the ultrasonic extraction method, the reaction is carried out at 30 to 50° C. for 0.5 to 2.5 hours, and the extraction solvent is water or 50 to 100% alcohol having 1 to 6 carbon atoms. Specifically, extraction is performed at 40 to 50° C. for 1 to 2.5 hours, and the extraction solvent is water or 70 to 80% alcohol having 1 to 6 carbon atoms.

The extraction solvent may be used in an amount of 2 to 50 times, more specifically, 2 to 20 times, based on the weight of the sample. For extraction, the sample may be left for 1 to 72 hours for leaching in the extraction solvent, and more specifically, for 24 to 48 hours.

After extraction, the extract may be fractionated by sequentially applying a fresh fractionation solvent. The fractionation solvent used for fractionation is any one or more selected from the group consisting of water, hexane, butanol, ethyl acetic acid, ethyl acetate, methylene chloride and mixtures thereof, preferably ethyl acetate or methylene chloride.

After obtaining the extract or fraction, a method such as concentration or freeze-drying may be additionally used.

A cosmetic composition according to another embodiment of the present invention is prepared by using the composition for improving atopic skin and moisturizing the skin.

The cosmetic composition of the present invention is in a form including the composition for improving atopic skin and moisturizing the skin. The cosmetic composition may be prepared in various forms according to a conventional cosmetic preparation method. For example, the cosmetic composition may be prepared in the form of cosmetic products, lotions, creams, lotions, etc. containing the composition for improving atopic skin and moisturizing the skin, which are diluted with a conventional cleansing solution, an astringent solution, and a moisturizing solution. can be used In addition, the cosmetic composition may include conventional adjuvants such as stabilizers, solubilizers, vitamins, pigments and fragrances commonly used in the field of cosmetic compositions.

A moisturizing cream according to another embodiment of the present invention is prepared using the composition.

A skin regeneration cream according to another embodiment of the present invention is prepared using the composition.

According to the composition for improving atopic skin and moisturizing the skin of the present invention, using calcined coral powder, Bupyeongcho and aloes, it improves atopic skin by showing the ability to inhibit the release of histamine and β-hexosaminidase of mast cells or neutrophils. It is possible to provide a composition for improving atopic skin and moisturizing the skin that strengthens the skin barrier and increases the moisture content.

1 is a graph of experimental results for cytotoxicity of calcined coral powder extract according to an embodiment of the present invention.
Figure 2 is a graph of the experimental results for the cytotoxicity of the extract of Bupyeongcho according to an embodiment of the present invention.
3 is a graph of the experimental results for the cytotoxicity of MIX1 mixed with calcined coral powder extract, Bupyeongcho extract, and aloesincense extract according to an embodiment of the present invention.
Figure 4 is a graph of experimental results for inhibition of nitric oxide (NO) production of calcined coral powder extract according to an embodiment of the present invention.
Figure 5 is a graph of experimental results for inhibition _{of PGE 2} production of calcined coral powder extract according to an embodiment of the present invention.
Figure 6 is a graph of the experimental results for the inhibition of TNF-α production of the calcined coral powder extract according to an embodiment of the present invention.
7 is a graph showing the results of IL-1β production of calcined coral powder extract according to an embodiment of the present invention.
8 is a Western blot analysis result for the decrease in the expression of cytokines according to the calcined coral powder extract treatment according to an embodiment of the present invention.
9 is related to the test results for the inhibitory effect of histamine release of the composition for atopic skin alleviation and skin moisturizing (MIX1) according to an embodiment of the present invention.
10 relates to the test results for the inhibitory effect of the β-hexoamidase release of the composition for atopic skin alleviation and skin moisturizing (MIX1) according to an embodiment of the present invention.

Hereinafter, embodiments of the present invention will be described in detail so that those of ordinary skill in the art can easily carry out the present invention. However, the present invention may be embodied in many different forms and is not limited to the embodiments described herein.

[Preparation Example 1: Preparation of extract]

1. Preparation of Bupyeongcho Extract

Bupyeongcho was thoroughly washed in running water and then completely dried naturally. After the dried Bupyeongcho was pulverized with a mixer and leached for 48 hours at room temperature using 80% ethanol, the sample was filtered to prepare a Bupyeongcho extract (SE).

2. Preparation of other natural extracts

Using the same method as the production method of the Bupyeongcho extract (SE), aloes aloes extract (AE), gogwang tree extract (PE), nanti tree extract (UE) and serrata oleifera extract (CE) were prepared, and calcined coral powder was Purchase After leaching for 48 hours at room temperature using 80% ethanol in the same manner as in the production method of the Bupyeongcho extract (SE), the sample was filtered to prepare a calcined coral powder extract (PE).

3. Preparation of Complex Extracts

The calcined coral powder extract (PE), Bupyeongcho extract (SE), aloes root extract (AE), gogwang tree extract (PE), nanthu tree extract (UE), and banyan tree extract (CE) are within the weight range as shown in Table 1 below. It was mixed to prepare a composition for improving atopic skin and moisturizing the skin.

MIX1 MIX2 MIX3 MIX4 MIX5 MIX6 PE 100 100 100 100 100 100 SE 100 100 100 100 100 100 AE 100 100 100 100 100 100 PE - 30 40 50 60 70 UE - 30 40 50 60 70 CE - 30 40 50 60 70

(Unit: parts by weight)

[Experimental Example 1: Anti-inflammatory effect]

Cell culture and samples

For the cell line, basophilic granulocytes RBL-2H3 and KU812 cells were distributed from the Korea Cell Line Bank, and 10% feta bovine serum (FBS, Hyclone, South Logan, UT, USA) and 1% antibiotics (100 U/ml penicillin G, 100 ㎍ / ㎖ streptomycin, Hyclone) was added DMEM medium was cultured at 37 ℃, 5% CO2 humidified conditions.

Cytotoxicity assessment

MTT analysis was performed using the MTT assay, which is an alternative animal test method suggested by the Ministry of Food and Drug Safety and the test standard presented by the Ministry of Food and Drug Safety. RBL-2H3 cells cultured in a cell culture dish were separated with Trypsin, mixed with a new medium, counted using a counting chamber and a microscope, and then dispensed at a concentration of 1 to 105 cells/ml in 96 well plates, respectively. Samples were prepared by concentration and cultured for 24 hours.

Cell line RBL-2H3 cells were dispensed and cultured for 24 hours, then calcined coral powder extract (PE) was treated for each concentration. After removing the medium, MTT solution was added at a concentration of 1 μg/ml and reacted at 37° C. for 4 hours. The MTT solution was removed and the reaction product formed by dispensing 100 μl of DMSO was dissolved, and after reacting for 30 minutes, absorbance at 540 nm was measured with an ELISA Reader.

The viability for each cell was compared and analyzed by expressing the difference in absorbance with respect to the control in percentage.

Cell viability (%) = [(absorbance of the sample added group)/absorbance of the non-added group] X 100

1 and 2 are the results of confirming the cytotoxicity of calcined coral powder extract (PE) and Bupyeongcho extract (SE) in RBL-2H3 cells, the cell viability measurement result, the concentration was increased, but there was little difference in the viability.

Figure 3 is a result of confirming the cytotoxicity of MIX 1 mixed with calcined coral powder extract (PE), Bupyeongcho extract (SE) and aloes hyang extract (AE), there was little difference in the survival rate.

NO production inhibition activity experiment

As a first step to measure the anti-inflammatory effect of the calcined oxygen powder extract (PE) at each concentration on the inflammatory response, NO in the cell culture medium was quantified and compared. To measure NO scavenging activity, RBL-2H3 cells were dispensed in a 96 well plate at 2x10 ⁴ cells/ml and cultured for 24 hours at 37°C, 5% CO ₂ conditions, and then calcined oxygen powder extract (PE) was added by concentration un, con 50 μg/ml, 100 μg/ml, 150 μg/ml, 200 μg/ml, 250 μg/ml).

To measure the amount of NO produced in the cell culture medium using Griess reagent, Griess reagent A was mixed with 35 ml of distilled water (DW), 15 ml of 100% acetic acid, and 0.5 g of sulfanilamide. Grease reagent B was prepared by adding 20 ml of DW, 30 ml of 100% acetic acid and 0.05 g of N-(1-Naphthyl)ethylenediamine.

After mixing 100 μl of cell culture supernatant and 100 μl of Griess reagent and reacting at room temperature for 10 minutes, absorbance was measured at 540 nm, which is the form present in the cell culture solution, and the amount of NO generated is sodium nitrite (NaNO ₂ ) compared with the standard.

As can be seen from the results of Figure 4, it was confirmed that the higher the concentration of the calcined oxygen powder extract (PE), the higher the NO production inhibitory activity.

Inflammatory cytokines and PGE ₂ production inhibitory activity experiment

Cells under the above conditions to verify the effect of oxalic acid powder extract (PE) on the production of inflammatory cytokines such as Prostaglandin E2 (PGE _{2), Interleukin (IL)-1β, and Tumor Necrosis Factor-α (TNF-α).} After culturing, the supernatant was collected and _{the contents of PGE 2} , IL-1β, and TNF-α (Enzo LifeScience) were quantified using an ELISA kit.

As can be seen from the results of Figures 5, 6 and 7, it was confirmed that the calcined oxygen powder extract (PE) significantly _{inhibited PGE 2} , TNF-α and IL-1β.

Confirmation of expression changes of iNOS, COX-2, TNF-α, and IL-1β through Western blot analysis. Cells were subjected to the above conditions to observe changes in the expression of transcription factors related to immune regulation at the protein level. After incubation, Western blot analysis was performed.

Prepared cells were treated with a lysis buffer containing 250 mM NaCl, 25 mM Tris-HCL (pH 7.5), 10 mM ethylenediaminetetraacetic acid, 1% Nonidet P-40, 0.1 mM phenyl-methysulfonylfuoride, and protease inhibitor. was dissolved. The concentration of the isolated protein was measured using a protein quantification reagent (Bio-Rad, Herculs, CA, USA). Equal amounts of proteins were separated using 10% sodium dodecy sulfate (SDS)-polyacrylamide gel electrophoresis and transferred to a nitrocellulose membrane (Schleicher % Schuell, Keene, NH, USA). Each membrane (membrane) was treated with 5% skim milk for 1 hour to block non-specific proteins.

Corresponding antibodies and enhanced chemiluminescence (ECL, Amersham Corp. Arlington Heights, IL, USA) solution was treated and then photosensitized with an X-ray film to confirm the protein expression level. The primary antibody used in this experiment was obtained from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA) and Cell signaling (Beverly, MA, USA), and the secondary antibody was purchased from Amersham Life Science Co. and used.

As a result, as can be seen in FIG. 8 , it was confirmed that NO production in RBL-2H3 cells was decreased by the treatment with the calcined oxygen powder extract (PE). It was confirmed that this decrease was due to a decrease in the expression of imducible NOS (iNOS).

Therefore, it will be said that calcined oxygen powder extract (PE) in RBL-2H3 cells exhibits anti-inflammatory effects due to reduced production of NO, PGE2, IL-1β, and TNF-α.

[Experimental Example 2: Atopic Skin Improvement Effect]

1. Measurement of histamine secretion

KU812 cells cultured in a cell culture dish were placed in a 50ml Conical Tube, centrifuged at 200xg speed for 5 minutes, the supernatant was discarded, and 10ml of tyrode buffer (pH7.2) was added and mixed. After centrifugation again at 200xg speed for 5 minutes, the supernatant was discarded, 30ml Tyrode buffer (pH7.2) was put into a 50ml conical tube, and inverting and pipetting were performed. After making 24 wells, the MIX 1 sample was treated with the test concentration and incubated for 15 minutes in a CO2 incubator.

After treatment with 2 μM A23187 in the control group and the test group treated with MIX 1 by concentration, 1ng/mL PMA was immediately treated and incubated for 30 minutes. After pipetting twice, 1ml supernatant was added to 15ml conical tube and centrifuged at 400xg speed for 5 minutes. 800μl of the supernatant was placed in a new 15ml conical tube, 1N NaOH, 1.5ml n-butanol, and 1ml chloroform were added, mixed for 5 minutes, and centrifuged at 200xg speed for 5 minutes. After that, 800 μl of the lower layer was added to a 15ml conical tube and 5ml of 1 NaOH was added to react. After centrifugation at 200xg speed for 5 minutes, it was added to a new conical tube, and 450μl 0.1N HCl and 1500μl n-heptane were added, followed by centrifugation for 1 minute. Then, 800μl from the lower layer was put into a new 15ml conical tube, 160μl 1N NaOH and 40μl ortho-phthalaldehyde were added, reacted for 4 minutes, 80μl 3N HCl was added, and the reaction was terminated. Then, 100 μl of each was added to the black plate, and the fluorescence intensity was measured at excitation350/365/emission465/465.

Mast cell degranulation is an important process for releasing histamine, and the released histamine is known as a mediator that induces allergic reactions such as itch and rash. The effect was verified. Histamine secretion was measured and evaluated in the blank test group A23187 treated with saline only, the control group treated with PMA, and the test group treated with A23187 and PMA after pretreatment with MIX 1 by concentration.

As shown in FIG. 9 , the histamine secretion was increased about 3 times in the control group that stimulated histamine secretion compared to the blank test group, and the histamine secretion stimulated by A23187 and PMA in the MIX 1 treatment group was suppressed and restored to normal. These results indicate that MIX 1 inhibits allergic reactions or has anti-atopic effects.

2. Hexoamidase secretion measurement

The RBL-2H3 cell line was aliquoted into a 24-well plate so that 2 × 105 cells per well were contained, and then sensitized with 200 ng/mL of anti-DNP Ig E per well and incubated for 12 hours in a 5% CO2 incubator. . After washing the cell line twice with siraganian buffer, MIX 1 samples were added by concentration and reacted again at 37°C for 30 min. This was treated with DNP-HAS (25 ng/mL), and after inducing an allergic reaction for 30 min, the reaction was stopped for 10 min in an ice bath, followed by centrifugation at 12,000 rpm for 3 min. Only the supernatant was recovered and used for beta-hexosaminidase measurement. To measure the inhibition of secretion of this enzyme, a reaction mixture of 30 μL of the supernatant and 30 μL of 1 mM p-NAG was reacted at 37° C. for 1 hour, the reaction was terminated by adding 250 mL of 0.1M carbonate buffer, and a microplate reader (Molecular Devices) , USA) was used to measure the absorbance at 405 nm. Histamine content was expressed by converting the absorbance ratio of the sample addition group and the control group to a % value.

As a result, in order to investigate the effect of MIX 1 on the anti-allergic effect, the secretion of β-hexosaminidase, which is known as a degranulation indicator of cells during an allergic reaction, was measured using the RBL-2H3 cell line, which is a major cell that exhibits an allergic reaction. As shown in FIG. 10 , the stimulated control group, which was sensitized with anti-DNP Ig E and treated with DNP-HAS to induce an allergic reaction, increased histamine secretion by 40% compared to the blank test group, and in the MIX 1 treatment group, anti-DNP Ig After sensitization with E, β-hexosaminidase activity stimulated by DNP-HAS was inhibited and restored to normal. These results indicate that MIX 1 inhibits allergic reactions or has anti-atopic effects.

[Experimental Example 3: Effect of complex extract]

1. Anti-inflammatory effect

As described above, the anti-inflammatory activity test results for the calcined oxygen powder extract (PE) were confirmed. Therefore, the anti-inflammatory activity test for the complex extracts MIX1 to MIX6 was conducted, and for the relative evaluation, the anti-inflammatory degree of the calcined oxygen powder extract (PE) was set as index 5, and the degree of the anti-inflammatory effect on the other complex extracts as the index described.

Anti-inflammatory activity test results for the complex extract are shown in Table 2 below.

PE MIX1 MIX2 MIX3 MIX4 MIX5 MIX6 Inhibition of NO production 5 6 6 8 9 10 7 Inhibition of inflammatory cytokine production 5 6 7 8 10 10 6 Inhibition of production of PGE ₂ 5 7 7 7 9 9 7

(Unit: Index)

As shown in Table 2, for the inhibition of NO production, inhibition of the production of inflammatory cytokines, and the _{inhibition of the production of PGE 2} for confirming the anti-inflammatory effect, relative comparison results to the calcined oxygen powder extract (PE), MIX1 to MIX6 It was confirmed that it exhibited an excellent anti-inflammatory effect equal to or greater than in

In particular, MIX3 to MIX6, which further include a gogwang tree extract (PE), an oleifera extract (UE), and an oleifera extract (CE), exhibited excellent anti-inflammatory effects.

2. Atopic skin improvement effect

As described above, the results of measuring histamine secretion and hexoamidase secretion for the calcined oxygen powder extract (PE) were confirmed. Accordingly, histamine secretion and hexoamidase secretion were measured for the complex extracts MIX1 to MIX6, and for a relative evaluation, the degree of inhibition of histamine and hexoamidase secretion of the calcined oxygen powder extract (PE) was set to index 5, and the other The degree of effect on the complex extract was described as an index.

The effect on the complex extract is shown in Table 3 below.

PE MIX1 MIX2 MIX3 MIX4 MX5 Histamine secretion inhibitory effect 5 7 8 9 10 6 Inhibitory effect of hexoamidase secretion 5 7 8 10 10 7

(Unit: Index)

As shown in Table 3 above, as a result of confirming the atopic skin improvement effect, it was confirmed that MIX1 to MIX6 exhibited equal or more histamine and hexoamidase secretion inhibitory effects.

In addition to the anti-inflammatory effect, MIX3 to MIX6, which further contains a high-gwang tree extract (PE), an oleifera extract (UE), and a serrata extract (CE), exhibited an excellent effect on atopic skin improvement effect.

[Experimental Example 4: Skin Moisturizing Effect]

For 10 women aged 20 to 25, the compositions MIX1 to MIX6 for improving atopic skin and moisturizing the skin were prepared as moisturizing creams, and the skin moisturizing effect was evaluated.

The test subjects used the moisturizing cream after washing their face twice every morning (between 7:00 and 9:00) and in the evening (between 7:00 and 9:00) every 12 hours for 4 weeks. At the 4th week, the moisturizing effect of the skin was evaluated by measuring the skin moisture evaporation rate.

The skin moisture evaporation rate was measured with Tewamter TM300 and was expressed as an average value of 10 subjects, and the results are shown in Table 4.

PE MIX1 MIX2 MIX3 MIX4 MIX5 MIX6 Water evaporation rate (g/h/m ² ) 17.23 18.24 19.77 20.22 22.09 23.49 19.34 Decrease rate of water evaporation compared to before first use (%) 39.16 35.59 30.19 28.50 26.12 27.55 34.24

As shown in Table 4, when the compositions MIX1 to MIX5 for atopic skin improvement and skin moisturizing prepared in the Preparation Example of the present invention are prepared as a moisturizing cream and continuously used, it can be confirmed that the skin moisture evaporation rate is gradually reduced. there was.

In particular, in the case of the composite compositions MIX3 to MIX5, even after 12 hours have elapsed after application, it was confirmed that the effect was significantly reduced compared to the moisture evaporation rate of the skin before use.

Formulation Example 1. Moisturizing cream

A moisturizing cream containing the MIX1 to MIX5 was prepared in a conventional manner according to the following composition components and composition ratios.

MIX1 3.0 wt%

8.0 wt% glycerin

Butylene glycol 5.0 wt%

Glyceryl Stearate / PEG-100 Stearate 1.0 wt%

Polysorbate 60 0.5 wt%

0.5 wt% sorbitan stearate

Phytosqualane 5.0 wt%

jojoba oil 3.0 wt%

15.0 wt% caprylic/capric triglycerides

Aloe Butter 2.0 wt%

beeswax 1.0 wt%

0.5% by weight of stearyl alcohol

0.5% by weight of beheryl alcohol

0.5% by weight of stearic acid

0.2 wt% of carboxyl vinyl polymer

0.1% by weight of xanthan gum

Arginine 0.5 wt%

Dimethicone 0.5 wt%

Beta glucan 0.2 wt%

0.5% by weight of preservative

Purified water remaining

Formulation Example 2. Moisturizing Cream

[Experimental Example 5: Atopic Treatment Effect Experiment]

For healthy females and males in their infancy to teens, 20s, and 30s suffering from atopic dermatitis, 10 people are randomly selected per each composition (MIX1 to MIX6) for improving atopic skin and moisturizing skin of the present invention, and atopic dermatitis One site where the lesion occurred was selected and MIX1 to MIX6 were applied to that site twice a day in the morning and in the evening, for 3 months. For the determination of the therapeutic effect of atopic dermatitis, after using the composition of the present invention, the degree of inhibition of gray hair generation was evaluated through the following evaluation scores as a subjective evaluation of each subject. The results are shown in Table 5 below.

number of people 0 points 1 point 2 points 3 points MIX 1 5 4 One - MIX 2 4 5 One - MIX 3 - One 3 6 MIX 4 - One 2 7 MIX 5 - - 3 7 MIX 6 - 3 3 4

(Unit: Index)

evaluation score

3: The atopic site after application was reduced to 90% or less compared to the atopic site before the start of application.

2: The atopic site after application was reduced to 60% or less compared to the atopic site before the start of application.

1: Compared to the atopic site before the start of application, the atopic site after application was reduced to 30% or less.

0: Compared to the atopic site, the atopic site after application did not decrease at all.

The treatment effect of atopic dermatitis according to the present invention is higher than that of MIX2 to MIX6 when mixing calcined coral powder extract (PE), Bupyeongcho extract (SE) and aloes root extract (AE) like MIX1, such as MIX2 to MIX6. It was confirmed that a synergistic effect appears when the complex treatment further includes (UE) and sagebrush extract (CE). In particular, in the case of MIX3 to 5, it was found that the effect was quite excellent.

Although the preferred embodiment of the present invention has been described in detail above, the scope of the present invention is not limited thereto, and various modifications and improvements by those skilled in the art using the basic concept of the present invention as defined in the following claims are also provided. is within the scope of the

Claims (8)

Calcined coral powder extract, Bupyeongcho extract, aloes hyang extract, gogwang tree extract, nanti tree extract and
A composition for improving atopic skin and moisturizing the skin. The method of claim 1,
The atopic skin is a symptom selected from the group consisting of dry skin, pruritus, dermatosis, and eczematous lesions.
A composition for improving atopic skin and moisturizing the skin. The method of claim 1,
The composition exhibits the alleviating effect of atopic skin through inhibition of histamine and β-hexosaminidase
A composition for improving atopic skin and moisturizing the skin. The method of claim 1,
The extract is extracted using an extraction solvent selected from the group consisting of water, C ₁ to C _{6 lower alcohols and mixtures thereof.}
A composition for improving atopic skin and moisturizing the skin. A cosmetic composition comprising the composition for improving atopic skin and moisturizing the skin according to any one of claims 1 to 4. A functional food composition comprising the composition according to any one of claims 1 to 4. A moisturizing cream comprising the composition according to any one of claims 1 to 4. A rejuvenating cream comprising the composition according to any one of claims 1 to 4.

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