KR20190075850A - A composition for improving, preventing and treating of pruritus comprising Porphyra yezoensis extract - Google Patents

Abstract

The present invention relates to pruritus using a laver extract as an active component. More specifically, the laver extract of the present invention inhibits the expression of pruritus-related genetic materials or proteins (MDC, TARC, ET-1, and PAR-2), thereby being able to be used for treating pruritus.

Description

FIELD OF THE INVENTION [0001] The present invention relates to a composition for improving, preventing or treating pruritus containing an extract of Kimchi, which comprises Porphyra yezoensis extract,

More particularly, the extract of the present invention inhibits the expression of an itch-related genetic substance or protein (MDC, TARC, ET-1, or PAR-2) Lt; / RTI >

Modern people are exposed to various environmental factors such as changes in dietary habits caused by rapid urbanization, pollution, skin dryness, house dust mites, animal hair and secretions, . Itching (pruritus) is defined as an unpleasant sensation that causes a desire to scratch or rub the skin (Andersen HH et al., Human surrogate models of histaminergic and non-histaminergic itch, Acta Dermato-Venereologica 95 (7): 771 7 (2015)), although the symptoms are common in skin diseases and systemic diseases, their characteristics are not well known.

Itching may be induced or even increased by various stimuli including physical, mechanical, and chemical factors. Inflammatory mediators also cause itching in several inflammatory skin diseases. However, not all forms of itching are associated with mediators, mechanical or electrical stimuli, and itching caused by dry skin may appear irrelevant to mediators.

To date, it has been shown that itching may be caused by histamine, Serotonin, Prostaglandin E, Tachykinin, Cytokines, Protease, opioid peptides, platelet-activating factor, and so on.

Histamine is synthesized in mast cells of the skin and stored in mast cell granules and secreted in response to various stimuli. H1 and H2 receptors are all present in the skin, but histamine causes itching through H1 receptors. Histamine is involved in many inflammatory skin diseases, including UV-induced dermatitis.

Serotonin is present in platelets and activates dermal mast cells to release histamine or to act on the 5-HT3 receptor of the central nervous system, causing itching.

Prostaglandin E itself does not cause itching, but it has the property of increasing itching by other mediators. Prostaglandin E is increased in skin diseases associated with itching, such as eczema and inflammation from ultraviolet light.

P substance (Substance P) is a neuropeptide of the tachykinin family. It is synthesized in the dorsal root ganglion of the C nerve fiber and is transported to the anhydrous sensory nerve fiber as a granule. It is a neurotransmitter . P substance is a potent vasodilator that causes redness and itching. P substance is released by trypsinase in mast cells and causes itching by liberating histamine in dermis mast cells either directly or through the NK-1 receptor.

Cytokines are low molecular weight proteins that are produced in all eukaryotic cells and act as cell surface receptors, including interleukin (IL), chemokines, and interferons. Although IL-1 and IL-8 do not cause itching, human IL-2 induces severe itching accompanied by skin eruptions and eosinophilia in cancer patients for therapeutic purposes.

One protease, kallikrein, causes itching when injected into the skin. Chymotrypsin, trypsin, and papain, which are proteases other than calicaine, also cause itching.

Opioid peptides cause itching by acting on the central or peripheral nervous system. Opioid peptides have three types of μ, δ and κ, and most pharmacological actions are inhibited by naloxone, so they can act through μ receptors. Morphine, when injected into the spinal cord, causes itching in the majority of patients and causes itching and redness irrespective of prostaglandin or histamine release during intradermal injection.

The platelet activating factor is a strong inflammatory inducer secreted from a wide range of inflammatory cells. The platelet activating factor has been reported to cause itching directly in animal experiments, but induces histamine secretion in mast cells in the human body and induces itching indirectly.

Causes of itching are classified into 1) seizure-prone itching, 2) winter itching, anal itching, vulvar itching, scrotum itching, watery itching, skin itching including scalp itching, 3) , Itching accompanied by an internal medical condition including malignant tumor, iron deficiency anemia, intracellular hypertension, hypothyroidism and hypothyroidism, diabetes mellitus, acquired immunodeficiency syndrome, chronic itching, itching rash, hairy wall, 5) mucus itching accompanied by other common colds and itching, eye pruritus associated with ophthalmic diseases such as conjunctivitis, dental anomalies accompanied with dental diseases, My itch.

In the case of steroids, it is absorbed to the blood vessels after passing through the skin layer, and when used for a long time, it causes skin irritation, capillary dilatation, and other abnormalities such as capillary dilatation And there is a risk of inducing adrenal suppression. Therefore, it is urgently required to develop a new therapeutic agent capable of preventing, alleviating, and treating itch with relatively few side effects.

Therefore, there is a need to develop a therapeutic agent for the prevention and treatment of new itching with no side effects and excellent therapeutic effect.

It should be understood that the foregoing description of the background art is merely for the purpose of promoting an understanding of the background of the present invention and is not to be construed as adhering to the prior art already known to those skilled in the art.

Patent Document 1: Korean Patent Laid-Open Publication No. 1999-0083931

The present inventors have made an effort to discover novel drugs having excellent stability that can inhibit the expression of inflammatory-related genetic material and protein, which are the causes of itching. As a result, the inventors of the present invention completed the present invention by confirming that MDC, TARC, ET-1, PAR-2 gene or protein expression is inhibited in human keratinocytes using a Kim extract.

Accordingly, it is an object of the present invention to provide a pharmaceutical composition for preventing or treating itching which contains an extract of Kim as an active ingredient.

Another object of the present invention is to provide a food composition for improving or preventing itching which contains an extract of Kimchi as an active ingredient.

Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

According to one aspect of the present invention, there is provided a pharmaceutical composition for preventing or treating itching comprising an extract of Kim as an active ingredient.

It is known that high concentrations of TSLP and macrophage-derived chemokine (MDC) are present in keratinocytes of patients suffering from itching. TSLP stimulates CD11c + dendritic cells to increase TARC (thymus and activation-regulated chemokine) and MDC production. These cytokine concentrations are closely related to the symptoms of itching, indicating that TARC and MDC are reliable biomarkers that best reflect the severity of itching.

In addition, the ET-1 protein is known to play a role in contracting blood vessels and raising blood pressure in the organs of the body, but it is an itch-related protein expressed independently of histamine-related itching (pruritus) in the skin. PAR-2 is a protein known as G-protein coupled receptor 11, which acts as a sensor for the activity of proteases, usually produced by inflammation and obesity. However, It is known as a protein that regulates pruritus. Therefore, ET-1 and PAR-2 are reliable biodegradable bio-barkers that can best reflect the itching caused by inflammation in skin tissue.

Accordingly, the present invention provides a method for inhibiting the proliferation of proliferating cells, which comprises administering a therapeutically effective amount of a compound selected from the group consisting of: itching caused by TSLP (thymic stromal lymphopoietin); itching caused by macrophage-derived chemokine; ET-1 (endothelin- And itching caused by various causes such as itching showed significant therapeutic efficacy.

Accordingly, the present inventors have found that inhibiting the expression of MDC, TARC, ET-1, and PAR-2, which are biomarkers showing itching symptoms such as MDC, TARC, ET-1 and PAR-2, ( Porphyra yezoensis ) was extracted with water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof, and the resultant extract was found to be atopic Thereby effectively inhibiting dermatitis.

More specifically, one aspect of the present invention relates to a pharmaceutical composition for preventing or treating itching comprising an extract of Kim, as an active ingredient.

Pyropia yezoensis is a seaweed of red algae, Kim peach, and has a length of 5 to 20 cm and a butterfly of 2 to 8 cm. The body has a leaf shape with one layer of cells, It is characterized by asexual reproduction and later sexual reproduction. In Korea, young foliage appears in October and grows to mature individuals until early December, and it may prosper from December to May of the following year.

The 'extract' in the present invention includes not only the crude extract obtained by treating the extraction raw material with the extraction solvent but also the crude extract extract. For example, the Kim extract can be prepared in powder form by an additional process such as vacuum distillation and lyophilization or spray drying.

The 'extract' in the present invention also includes fractions obtained by further fractionating the crude extract. Namely, the Kim extract includes not only the extract obtained by using the extraction solvent but also the one obtained by additionally applying the purification process thereto. For example, fractions obtained by passing through an ultrafiltration membrane having a constant molecular weight cut-off value and various other methods, such as separation by various chromatographies (produced for separation by size, charge, hydrophobicity or affinity) ≪ / RTI >

Also, in the present invention, the 'extract of Kimchi' is an extract of Kim, especially radish stain, with water, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof, and the extraction method is not particularly limited. Preferably, the Kim extract may be a hot water extract of Kimchi or an aqueous solution of Kim ethanol, for example, Kimchi extract.

The Kim extract may be used directly as an extract. Preferably, the Kim extract can be obtained by naturally drying or roots obtained from nature, in order to contain an excessive amount of active substance present in the kimchi. , Chitosan, and the like, followed by drying or pulverization, or by mixing the kimchi with acetic acid, treating the kimi with electron ionization energy, treating the kimi with microwave, Any method of fermenting into microorganisms can be used without limitation.

For example, in order to contain an excessive amount of the effective substance present in the kimchi, the kim was recovered and dried in the shade for 1 week or more, preferably 1 month or more, more preferably 3 months or more, Can be used.

When the water is used as the solvent, the Kim extract is preferably extracted once or twice or more in water at 40 to 105 ° C, preferably 90 to 100 ° C for 0.5 to 50 hours, preferably 1 to 6 hours . Even if no hot water is used as the solvent, the components of the Kim can be extracted from the diluted solution obtained by diluting Kim with cold water or room temperature water.

An extract of the above-mentioned lower alcohol having 1 to 4 carbon atoms can be used. For example, extraction is carried out with an alcohol aqueous solution such as ethanol, methanol or isopropanol, preferably an aqueous alcohol solution of 20 to 80% by weight, more preferably an aqueous alcohol solution of 80% by weight. When alcohol extracts are used, first concentrate or dry the alcohol concentrate or alcohol extract, which has lowered the alcohol content by evaporating alcohol from the alcohol extract, and then dissolve in water before use.

The solid content of the Kim extract is not particularly limited, but it is 1 to 15% by weight when there is no other concentration step after extraction. The extract may be used directly, or may be concentrated, or may be diluted by concentrating or drying.

The Kim extract did not exhibit cytotoxicity in cells treated with IFNγ and TNFα, which are inflammation inducers, as in the experiment described later. As a result, it can be seen that the extract is very stable even when it is used for skin which is weakened by itching. (MDC, TARC, ET-1, and PAR-2) associated with itching such as TARC, MDC, ET-1 and PAR-2 in cells that are pruritized by IFN-γ and TNF- 2) is suppressed and inhibited, the composition of the present invention is excellent in prevention or treatment of itch, and more preferably, it is effective in prevention, treatment or improvement of severe atopic dermatitis accompanied by itch . The above-mentioned Kim extract can be used as a liquid extract itself or it can be dried and powdered.

As used herein, the term " comprising as an active ingredient " is meant to include an amount sufficient to achieve an itch improvement, treatment or prophylactic efficacy or activity of the extract of the present invention.

In the present invention, " prevention " may mean all actions of inhibiting, delaying or alleviating itching by administering the composition according to the present invention to an individual. In the present invention, 'treatment' may mean all the actions that the composition according to the present invention is administered to suspected itching individuals so that the symptoms of itching may be improved or benefited.

The term "itching" or "pruritus" in the present specification is not particularly limited, and includes the following: paroxysmal itching, winter itching, anal itching, vulvar itching, scrotum itching, watery itching, scalp itching, nasal itching, Itching; Itching accompanied by an internal medical disorder such as gall bladder, chronic itching, chronic renal failure, malignant tumor, iron deficiency anemia, intrinsic erythropoiesis, hyperthyroidism, hypothyroidism, diabetes and acquired immunodeficiency; And itchy skin diseases such as chronic simple poisoning, itching rash, hairy walls, nervous scraping wounds, behavioral disorders involving the skin and parasitism, and itching associated with atopic dermatitis.

Paroxysmal itching is a seizure-prone itch, which is seen in chronic simple poisons or dermatitis.

Winter itching occurs in more than 50% of elderly people over 70 years of age, and should be differentiated from itching caused by small skin diseases such as omphalitis, squamous cell carcinoma, or systemic disease. Women may be symptomatic of postmenopausal syndrome. It is a major cause of skin dryness due to decreased moisture content of aged skin and gradual decrease of sebaceous secretion, and fine cracks and stains are mainly seen in the upper and tibial parts.

Anal anxiety is an uncomfortable feeling of scratching the skin around the anus, often involving psychogenic factors. It may occur regardless of age, but it occurs more often after middle age. However, the cause of all anal itching is not only psychogenic, but it can be caused by contamination and irritation around the anus. Colonic anomalies such as dentition, hemorrhoids, fistula, and chronic diarrhea, spicy foods, and medicines can further aggravate the irritation. Several infectious diseases such as Staphylococcus, Streptococcus, Fungus, Candida, and Herpes simplex virus can cause itching. Candida infections are most common, with cracks appearing at the time of infection and the epidermis appearing to be watery. It can cause severe itching even when the skin disease such as psoriasis, dermatitis dermatitis, or squamous cellulitis is in the anal region, and lesions can be observed in other parts. Neurodermatitis of the anus can be scratched by the affected part until it is bleeding due to severe itching, and it can show the same findings as the chronic simple popliteal part of the other part.

The most common cause of vulvar itching is Candida infection. Other factors may include trichomoniasis vaginitis, contact dermatitis due to pads, birth control pills, vaginal wash liquids, condoms, and the like. After middle age, cystic atrophy is a common cause. In Fox-Fordyce disease, severe itching may also occur. However, transient itching may also be caused by friction, sweating, or vomiting during pregnancy.

In relation to scrotum itching, adult scrotum has immunity to fungal infections like adult scalp, but it is a site where localized chronic simple poisoning occurs well. The cause is often a psychogenic factor, a severe fever, and intensive treatment It may last for many years.

Waterborne itching is severe discomfort with needle striking within a few minutes after exposure to water or after stopping exposure to water, and lasts for about an hour. It is independent of the temperature of the contacted water and no particular change is observed in the skin. For some patients, it may also be caused by changes in ambient temperature. About one-third of patients have family history and are usually chronic and unresponsive to treatment. Histamine is not the only mediator, given the increased levels of histamine in the skin and blood, but the symptoms are not alleviated by antihistamines. It is similar to the symptoms of intracerebral hemorrhage.

Scalp itching can occur as an independent symptom without obvious lesions of the scalp, and it can be seen in middle-aged or elderly people. Itching is very severe and appears to be spasmodic, which is exacerbated by fatigue or stress. Differential diseases include herpes dermatitis, chronic simplex poisoning, seborrheic dermatitis and psoriasis.

Patients with biliary cirrhosis are accompanied by severe systemic itching. Itching is associated with an increase in plasma bile acid concentrations and clinically itching can lead to severe itching when applied directly to blistered skin lesions at a concentration of bile acid.

About 20-50% of patients with chronic renal failure receiving hemodialysis receive itching. Itching may occur locally or systemically, and most of the time during hemodialysis, or hemodialysis may cause temporary symptom relief. It has been reported that there is no direct relationship between the levels of histamine, urea, and creatinine in the blood and the degree of itching. Some patients have dry skin, but most have normal skin and use of moisturizers does not relieve or reduce symptoms.

In relation to malignant tumors, extensive examination of malignant tumors is necessary if systemic itching occurs without a specific cause in middle age or old age. In 15-25% of patients with Hozikin (a typical disease that causes swelling of the lymph nodes), itching is persistent and sometimes accompanied by burning pain and burning, which is unknown. Systemic itching can also occur in leukemia.

Iron deficiency can also cause itching. It has been reported that oral administration of iron supplements to patients with acute erythropoiesis and iron deficiency resulted in a reduction of itching.

Approximately 50% of patients with severe erythropoiesis experience severe itching within minutes after contact with water, and these symptoms last for about 15 to 60 minutes. It usually occurs after bathing and is called bath itch. There is no specific change in skin and it occurs irrespective of water temperature. However, histamine is increased in serum and urine. Platelet aggregation is thought to be the cause of various itching mediators including histamine.

Severe systemic itching may occur in hyperthyroidism. Increased skin blood flow is caused by increasing skin surface temperature and lowering the threshold for itching. In hypothyroidism, the skin becomes severely dry during mucous scintigraphy and systemic itching may occur. In both diseases, itching of the genital area due to mucocutaneous candidiasis may occur.

In some diabetic patients, itching of the anal region due to mucocutaneous candidiasis may occur. However, some patients may have systemic itching.

One of the important symptoms of AIDS is itching. The causes of itching in AIDS patients are omnidia, diabetes, candidiasis, dermatitis, and systemic diseases such as kidney failure and cholestasis. Pigmented or pigmented rashes of the whole body, which characteristically cause severe itching, also occur.

Chronic simple poisonous is a skin-like disease that repeatedly rubs or rubs the skin. Itching may occur in normal skin, resulting in secondary chronic simple poisons. It usually occurs in the 30s to 50s, and is more common in women than in men.

Itchy rash is characterized by multiple nodules with severe itching and is not treated well and has long lasting characteristics. The cause is not well known and can be caused by anemia, liver disease, HIV disease, pregnancy, kidney failure, and mental stress.

The hair of the hair is a neurosis that pulls the hair by an abnormal desire. Mental, and social stress, stress in family, stress in school life, sense of competition among siblings, director, mother 's admission, maternity relationship, etc. It occurs in almost all age groups, from children to adults.

A nervous scraping wound is an iterative and compulsive disorder in which the skin of a person is torn, pinched, scraped and skin lesions occur. The patient acknowledges that his or her behavior has caused the lesion but can not inhibit it. It may occur at any age, but it is common in middle-aged women and may be caused by psychological stress. Itching, insect palsy, and other skin lesions. Neurotic scarring is also associated with depression, obsessive compulsive disorder, and anxiety. These symptoms are more likely to occur in people with perfectionism whose personality is compulsive, stubborn, controllable, and fearful of wrongdoing.

Artificial dermatitis is a dermatitis that is caused by artificially wounded skin to cause compassion or to avoid responsibility. Skin lesions are caused by mechanical methods, chemicals, caustics, and the like. Other causes include nails, sharp tools, and hot metal. The patient hurts his or her body to satisfy psychological needs, is more likely to occur in women and may appear in all age groups. The majority of patients have infantile, dependent, and impulsive personality disorders.

Behavioral disorders involving the skin can result in a variety of physical impairments due to long - term, repeated, obsessive behavior. The laceration that is given to oneself is made for the purpose of suicide, and sometimes it is attempted to show courage to puberty.

Parasitism is a disease in which the parasite is parasitic on the patient's skin. It is a single symptom of hypochondriasis, which is chronic, body-related delusions, without impairment of personality or thinking ability. Patients are asked to bring small epidermal debris into a small compartment, paper tissue, or tape to be inspected, and 2-3% of patients experience parasite infections.

Other it may be nasal itching accompanied with nose-related diseases such as rhinitis, eye pruritus accompanied with ophthalmic diseases such as conjunctivitis, and itching in mouth due to dental cause.

The types of itching to be prevented or treated according to the present invention are not limited, and may be severe itching with MDC, TARC, ET-1 and PAR-2 as biomarkers due to severe atopic dermatitis.

The food composition may be one prepared by formulating the Kim extract in the form of a capsule, tablet, powder, granule, liquid, ring, flake, paste, syrup, gel, jelly or bar, , Gum, confectionery, etc., and it is produced in the form of general food. When it is taken, it means bringing about a certain effect in health. However, unlike general medicine, There is no side effect that can be.

The food composition is very useful because it can be ingested routinely. The amount of the Kim extract added in such a food composition can not be uniformly determined depending on the type of food to be targeted but may be added within a range that does not impair the original taste of the food, %, Preferably 0.1 to 20 wt%. In the case of foods in the form of capsules, tablets, powders, granules, liquids, pellets, flakes, pastes, syrups, gels, jellies or bar, the content is usually 0.1 to 100% by weight, preferably 0.5 to 80% .

The food composition may contain, as an active ingredient, not only a Kim extract but also a component that is ordinarily added in food production, and includes, for example, proteins, carbohydrates, fats, nutrients, seasonings and flavors. Examples of the above-mentioned carbohydrates are monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And polysaccharides such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol.

Natural flavorings such as tau martin and stevia extract (e.g., rebaudioside A and glycyrrhizin) and synthetic flavorings (saccharine, aspartame, etc.) can be used as flavorings. For example, when the food composition of the present invention is prepared from a drink and a beverage, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, juice, various plant extracts and the like may be further added in addition to the extract of the present invention.

The food composition of the present invention includes human food, beverages, animal feed, and feed additives.

Furthermore, in the animal feed additive comprising the extract of Kim extract as an active ingredient, the extract may be prepared in powder form and may be further added to an animal feed to produce an animal feed additive comprising a polysaccharide have. The animal feed may be selected from the group consisting of animal powder feed, calf substitute oil, or porcine animal feed, and the content of the powdery extract is in the range of 1-2 g (0.1% level) per kg of feed Is sufficient.

In the pharmaceutical composition for the treatment or prevention of itch with the extract of Kim extract as an active ingredient, the extract of Kim extract may be used in an amount of 0.001 mg / kg or more, preferably 0.1 mg / kg or more, more preferably 10 mg / More preferably at least 100 mg / kg, even more preferably at least 250 mg / kg, most preferably at least 0.1 g / kg. Since the Kim extract has no adverse effect on the human body even when administered in an excessive amount as a natural product, the quantitative upper limit of the Kim extract contained in the composition of the present invention can be selected by a person skilled in the art within a suitable range.

The pharmaceutical composition may be prepared by using pharmaceutically acceptable and physiologically acceptable adjuvants in addition to the above-mentioned active ingredients. Examples of the adjuvants include excipients, disintegrants, sweeteners, binders, coating agents, swelling agents, lubricants, A flavoring agent and the like can be used. The pharmaceutical composition may be formulated to contain at least one pharmaceutically acceptable carrier in addition to the above-described effective ingredients for administration.

The pharmaceutical form of the pharmaceutical composition may be a granule, a powder, a tablet, a coated tablet, a capsule, a suppository, a liquid, a syrup, a juice, a suspension, an emulsion, a drip agent or an injectable liquid agent. For example, for formulation into tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Also, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included as a mixture. Suitable binders include, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, natural and synthetic gums such as corn sweeteners, acacia, tracker candles or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum and the like.

Acceptable pharmaceutical carriers for compositions that are formulated into a liquid solution include sterile water and sterile water suitable for the living body such as saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, And other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added as needed. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable solutions, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like.

Further, it can be suitably formulated according to each disease or ingredient, using the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA as an appropriate method in the field.

The pharmaceutical composition may be administered orally or parenterally. In the case of parenteral administration, the pharmaceutical composition may be administered by intravenous injection, subcutaneous injection, muscle injection, intraperitoneal injection, transdermal administration or the like, preferably oral administration.

The appropriate dosage of the pharmaceutical composition will vary depending on factors such as the formulation method, the manner of administration, the age, weight, sex, pathological condition, food, administration time, route of administration, excretion rate and responsiveness of the patient, The physician can easily determine and prescribe dosages effective for the desired treatment or prophylaxis. According to a preferred embodiment, the daily dosage of the pharmaceutical composition is 0.001-10 g / kg.

The pharmaceutical composition may be prepared in a unit dose form by formulating it using a pharmaceutically acceptable carrier and / or excipient according to a method which can be easily carried out by a person having ordinary skill in the art to which the present invention belongs. Into a capacity container. The formulations may be in the form of solutions, suspensions or emulsions in oils or aqueous media, or in the form of excipients, powders, granules, tablets or capsules, and may additionally contain dispersing or stabilizing agents.

The pharmaceutical composition of the present invention includes human and animal medicines. Specifically, the above-mentioned Kim extract can be used for manufacturing various veterinary pharmaceuticals. In this case, the veterinary medicine can be used as an animal medicine or an adjuvant for effectively treating or preventing an atopic dermatitis disease of an animal, an auxiliary therapeutic agent for enhancing therapeutic effect, or a feed additive.

For this purpose, the composition of the present invention may be formulated by itself or together with a carrier which is conventionally accepted in the pharmaceutical field, for oral administration such as tablets, capsules, liquids and suspensions which are customary in the pharmaceutical field Formulations, injectable preparations or suspensions, feed additives, and the like. In addition, in order to prevent the drug from being degraded by gastric acid during oral administration, a solid preparation for oral administration such as an antacid or a tablet may be formulated into an enteric coated fat preparation.

The dosage of the polysaccharide fraction of the extract according to the present invention to the human body should be appropriately used depending on the degree of absorption, inactivation rate and excretion rate of the active ingredient in the animal, the age, sex and condition of the animal, kg to 100-400 mg per day.

Yet another aspect of the present invention relates to a cosmetic composition for improving or preventing itching comprising an extract of Kim as an active ingredient. Preferably, MDC, TARC, ET-1, and PAR-2 due to severe atopic dermatitis may be caused by the expression of MDC, TARC, ET-1 and PAR- Which may be severe itching.

The cosmetic composition for improving or preventing itching may be at least one selected from the group consisting of ointment for external use, cream, softening longevity, nutritional lotion, pack, essence, hair tonic, shampoo, rinse, hair conditioner, hair treatment, gel, skin lotion, Nourishing cream, Eye cream, Moisturizing cream, Hand cream, Mask pack, Foundation, Nutrition essence, Sunscreen, Soap, Cleansing foam, Cleansing lotion, Cleansing cream, Lotion, Milk lotion, Moisturizing lotion, Nourishing lotion, Massage cream , Body lotions, and body cleansers, but are not limited thereto. The composition of each of these formulations may contain various kinds of bases and additives necessary for formulation of the formulation, and the kinds and amounts of these ingredients can be easily selected by those skilled in the art.

When the formulation of the present invention is a paste, cream or gel, animal fibers, plant fibers, wax, paraffin, starch, tracant, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide are used as carrier components .

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

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

When 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, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component is selected from 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, linolenic derivatives or ethoxylated glycerol fatty acid esters.

The features and advantages of the present invention are summarized as follows:

(I) The present invention provides a pharmaceutical composition for the prevention or treatment of itching comprising an extract of Kim as an active ingredient.

(Ii) The present invention also provides a food composition for improving or preventing itching comprising an extract of Kim as an active ingredient.

(Iii) Since the composition of the present invention inhibits itching induced by the expression of MDC, TARC, ET-1 and PAR-2 and does not show cytotoxicity in inflammation-induced weakened cells, TARC, ET-1, and PAR-2.

FIG. 1A is a graph showing the cell survival rate of inflammatory HaCaT cells according to the concentration of the extract of Kimchi. FIG. 1A is a graph showing cell viability of HaCaT cells without inflammation induced by addition of a Kim extract will be.
FIG. 1B shows the cell viability of HaCaT cells induced by inflammation when the ethanol extract of Kim was added.
FIG. 1C shows the cell survival rate when the extract of Kim extract was added to inflammatory HaCaT cells.
FIG. 2 is a graph showing the results of measurement of itching-related gene expression in inflammation-induced HaCaT cells according to the concentration of ethanol extract of Kimchi. FIG. 2a shows MDC Gene expression pattern.
FIG. 2B shows the expression of MDC gene in HaCaT cells induced by TNFα, when the ethanol extract of Kim was added.
FIG. 2C shows the expression pattern of TARC gene upon addition of the ethanol extract of HaCaT to IFNγ-induced HaCaT cells.
FIG. 2d shows the expression of TARC gene when HaCaT cells induced inflammation with TNFα and added ethanol extract of Kim.
FIG. 3A shows the expression patterns of MDC protein when HaCaT cells that induced inflammation with IFNγ were added with Kim ethanol extract.
FIG. 3B shows the expression of MDC protein when HaCaT cells induced inflammation with TNFα and added ethanol extract. FIG.
FIG. 4A is a graph showing the ET-1 gene expression pattern when HaCaT cells induced inflammation with IFNγ added with the ethanol extract of Kim.
FIG. 4B shows the expression pattern of ET-1 gene when HaCaT cells that induced inflammation with TNFα were added with the ethanol extract of Kim.
FIG. 5A shows the expression pattern of PAR-2 gene when HaCaT cells that induced inflammation with IFNγ were added with the ethanol extract of Kim.
FIG. 5B shows the expression of PAR-2 gene when HaCaT cells that induced inflammation with TNFα were added with the ethanol extract of Kim.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

statistics

2 to 5 obtained in this experiment were statistically processed as follows. The data obtained from each experiment set the normalized expression rate to 1 in the untreated control group and the data showing significant differences after one-way ANOVA (α = 0.05) were obtained by Tukey's honestly significance difference [HSD]). ### P <0.001, control group versus IFN-γ or TNF-α (10 ng / mL) alone. * P <0.05, ** P <0.01, *** P <0.001 were considered to be statistically significant.

Example

Example 1 Preparation of Kim Ethanol Extract

Radiation pattern Kim (Porphyra yezoensis) was used and dried with Kim domestic radiation patterns (Porphyra yezoensis), cold storage.

In the Erlenmeyer flask, 10 g of the above-mentioned dried radish flesh and 100 ml of an 80% by weight ethanol aqueous solution were added, and the mixture was immersed and extracted twice at 45 ° C for 24 hours, filtered, concentrated by a vacuum concentrator and lyophilized, &Lt; / RTI &gt;

40 ml of distilled water was added to the completely concentrated extract of the extract to dissolve for one day, and the undissolved portion was removed by using a centrifuge. The supernatant was taken and dissolved in an organic solvent (chloroform / methanol / distilled water = 2: 1: 0.9, v / v), and the supernatant was separated by a centrifuge. The separated supernatant was dried with a freeze dryer to obtain final ethanol extract powder.

Thereafter, the ethanol extract of Kimchi was diluted with DMSO (dimethyl sulfoxide, Sigma, USA) so as to have a final concentration of 200 mg / ml, and the membrane filter paper (0.2 μm, Millipore, USA) was used in the experiment.

EXPERIMENTAL EXAMPLE 1 Analysis of Cell Survival According to Kim's Ethanol Extract-1 -

37 ℃, 5% under CO ₂ conditions, the human-derived keratinocytes in HaCaT cell line, respectively 1% penicillin-streptomycin (penicillin-streptomycin) and 10% fetal calf serum (fetal bovine serum) a DMEM (Dulbecco's Modified Eagle's Medium containing ) Medium (Gibco BRL, USA).

The HaCaT cell line cultured under the above conditions was dispensed into a 96-well plate or a 10-ml culture dish at 1 × 10 ⁵ cells / ml and cultured for 24 hours. Then, the medium was removed and a fresh medium was added.

In the control group, the ethanol extracts (1.56, 3.12, 6.25, 12.5, 25, 40, 50, and 50%) obtained in Example 1 were added to the cells in the experimental group, 100, 125, 200, 250, 500, 1000, 1500, 2000 占 퐂 / ml). The control and experimental groups were incubated for 24 hours and cell viability was measured.

In order to compare the cell survival rate according to the concentration of ethanol extract of Kimchi, each of the cells prepared in Experimental Example 1 was recovered and MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium solution (0.05 ㎎ / ㎖) was added and the reaction was induced by incubating in a cell incubator for 4 hours. Then, the MTT solution was removed and DMSO was added to completely dissolve the formazan crystal. 200 쨉 l of each well was added to each well and the absorbance was measured at 570 ㎚ using a microplate reader.

FIG. 1A is a graph showing the cell survival rate of inflammatory HaCaT cells according to the concentration of ethanol extract of Kim. FIG. 1A is a graph showing cell viability when HaCaT cells without inflammation were added with Kim ethanol extract Respectively.

As shown in FIG. 1A, no cytotoxicity was observed when HaCaT cell extract was treated with the ethanol extract of 1.56 to 1000 占 퐂 / ml. However, in the case of treatment with the ethanol extract of 2000 ㎍ / ㎖, the effect of the ethanol extract on the cell viability is weak because the cell survival rate is reduced by about 20%. However, in order to obtain higher stability, It is preferable to treat the ethanol extract of Kim.

<Experimental Example 2: Analysis of cell viability according to Kim's ethanol extract-2>

37 ℃, 5% under CO ₂ conditions, the human-derived keratinocytes in HaCaT cell line, respectively 1% penicillin-streptomycin (penicillin-streptomycin) and 10% fetal calf serum (fetal bovine serum) a DMEM (Dulbecco's Modified Eagle's Medium containing ) Medium (Gibco BRL, USA).

The HaCaT cell line cultured under the above conditions was dispensed into a 96-well plate or a 10-ml culture dish at 1 × 10 ⁵ cells / ml and cultured for 24 hours. Then, the medium was removed and a fresh medium was added.

In the control group, IFN-y (10 ng / ml) and the ethanol extract of Kimchi (1.56, 3.12, and 6.25) obtained in Example 1 were added to each of the cells, , 12.5, 25, 40, 50, 100, 125, 200, 250, 500, 1000, 1500, 2000 占 퐂 / ml). The control and experimental groups were incubated for 24 hours and cell viability was measured.

In order to compare the cell viability according to the concentration of the ethanol extract of Kimchi, each cell prepared in Experimental Example 2 was collected and then MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl tetrazolium solution (0.05 ㎎ / ㎖) was added and the reaction was induced by incubating in a cell incubator for 4 hours. Then, the MTT solution was removed and DMSO was added to completely dissolve the formazan crystal. 200 쨉 l of each well was added to each well and the absorbance was measured at 570 ㎚ using a microplate reader.

FIG. 1B shows the cell survival rate when the ethanol extract of Kimchi HaCaT was added to the inflammation-inducing HaCaT cells. According to the results, even in the inflammation-induced cells treated with IFN-y, the inflammation inducer, The extract (1.56-1000 占 퐂 / ml) was confirmed to have no cytotoxicity. Therefore, even if the cells treated with the ethanol extract of inflammation do not cause cytotoxicity, they would be very stable.

<Experimental Example 3: Analysis of cell viability according to Kim's ethanol extract-3>

37 ℃, 5% under CO ₂ conditions, the human-derived keratinocytes in HaCaT cell line, respectively 1% penicillin-streptomycin (penicillin-streptomycin) and 10% fetal calf serum (fetal bovine serum) a DMEM (Dulbecco's Modified Eagle's Medium containing ) Medium (Gibco BRL, USA).

The HaCaT cell line cultured under the above conditions was dispensed into a 96-well plate or a 10-ml culture dish at 1 × 10 ⁵ cells / ml and cultured for 24 hours. Then, the medium was removed and a fresh medium was added.

In the control group, TNF-α (10 ng / ml) and the ethanol extract of Kimchi (1.56, 3.12, and 6.25) obtained in Example 1 were added to each cell, , 12.5, 25, 40, 50, 100, 125, 200, 250, 500, 1000, 1500, 2000 占 퐂 / ml). The control and experimental groups were incubated for 24 hours and cell viability was measured.

In order to compare the cell viability according to the concentration of the ethanol extract of Kimchi, each of the cells prepared in Experimental Example 3 was collected, and then MTT (3- (4,5-dimethylthiazol-2-yl) tetrazolium solution (0.05 ㎎ / ㎖) was added and the reaction was induced by incubating in a cell incubator for 4 hours. Then, the MTT solution was removed and DMSO was added to completely dissolve the formazan crystal. 200 쨉 l of each well was added to each well and the absorbance was measured at 570 ㎚ using a microplate reader.

FIG. 1C shows the cell survival rate when the ethanol extract of Kimchi was added to the inflammatory HaCaT cells. According to the results, even in the inflammation-induced cells treated with the inflammation inducer TNF-a, The extract (1.56-1000 占 퐂 / ml) was confirmed to have no cytotoxicity. Therefore, even if the cells treated with the ethanol extract of inflammation do not cause cytotoxicity, they would be very stable.

Taken together, it can be seen that the Kim ethanol extract of the present invention is a very stable substance that does not show cytotoxicity against all of HaCaT cells in which inflammation occurs or does not occur.

EXPERIMENTAL EXAMPLE 4: Analysis of the expression level of the itch-related gene [

(MDC and TARC) expression in HaCaT cells induced by inflammation was investigated by the addition of P. yezoenisis extracts at various concentrations (40, 200, 1000 ㎍ / ㎖).

The gene was analyzed using RT-PCR for analysis. Specifically, IFN-γ or TNF-α, which is an inflammation inducer, was treated at 10 ng / ml, and at the same time, P. yezoenisis was treated with various concentrations (40, 200, 1000 ㎍ / After culturing for 24 hours, RNA was isolated by treatment with Trizol reagent and synthesized with complementary DNA using cDNA synthesis kit (TOYOBO, Japan). The synthesized cDNA was mixed with the prepared MDC, TARC, ET-1 or PAR-2 Primer (BIONEER, Korea) and hybridized with SYBR Green PCR Master Mix (BIO-RAD, , Heating at 95 ° C for 10 minutes, reaction at 95 ° C for 30 seconds, 60 ° C for 30 seconds, and 72 ° C for 30 seconds for 40 cycles. When the inflammation inducer and the ethanol extract were added, Gene expression patterns of itch-related factors were analyzed.

FIG. 2 is a graph showing the results of measurement of itching-related gene expression in inflammation-induced HaCaT cells according to the concentration of ethanol extract of Kimchi. FIG. 2a shows MDC FIG. 2B shows the results of measurement of the MDC gene expression pattern when HaCaT cells induced inflammation with TNFα, and FIG. 2C shows HaCaT cells induced by inflammation with IFNγ FIG. 2d shows the results of measurement of TARC gene expression upon addition of the ethanol extract to HaCaT cells that induced inflammation with TNFα, and FIG. 2d shows the expression of TARC gene when the ethanol extract of Kimchi 4a shows the expression pattern of ET-1 gene when Haemaphylococcus aureus extract was added to HaCaT cells that induced inflammation with IFNγ FIG. 2d shows the expression pattern of PAR-2 gene upon addition of the ethanol extract of HaCaT cells induced by TNFα.

As shown in FIG. 2, the expression of MDC and TARC gene related to atopic dermatitis was dose-dependently induced by treatment with ethanol extract of Kimchi (40, 200, 1000 ㎍ / ㎖) in HaCaT cells that induced inflammation with IFNγ Respectively.

In addition, the expression of TARC gene was significantly decreased in HaCaT cells induced by TNFα by ethanol extract (40, 200, 1000 ㎍ / ㎖).

However, in the HaCaT cells induced by TNFα, MDC gene expression was decreased most when 40 ng / ml of ethanol extract was treated with 200 ng / ㎖ of ethanol extract, But the amount of MDC gene expression was decreased in HaCaT cells treated with TNFa alone, although it was slightly lower when treated with 1000 ng / ml ethanol extract of Kimchi.

As shown in Figs. 4A and 4B, when ET-1 is significantly increased by stimulating skin keratinized cells (HaCaT cells) by treating IFN-y or TNF-a.

On the other hand, treatment of 40, 200, and 1000 ㎍ / ㎖ of ethanol extract of Kimchi after treatment with IFN-γ or TNF-α to stimulate dermal keratinocytes (HaCaT cells) significantly decreased the protein production of ET-1 Respectively.

In other words, the ethanol extract of the present invention can significantly reduce the expression of ET-1, a genetic substance related to itching (pruritus), which is independently expressed from histamine-related ischemia in the skin keratinocytes induced by inflammation Respectively.

As shown in FIGS. 5A and 5B, when the skin keratinized cells (HaCaT cells) are stimulated with IFN-γ or TNF-α, the skin barrier is damaged and the expression of PAR-2 protein is increased.

On the other hand, it was found that the production of PAR-2 protein was remarkably increased by treatment with IFN-γ or TNF-α to stimulate skin keratinocytes (HaCaT cells) and then treatment with 40, 200, 1000 μg / have.

That is, the ethanol extract of Kimchi of the present invention induces reduction of the skin barrier-related factor (PAR-2) induced by the inflammation reaction in the skin, and as a result, it has the effect of improving the skin itching.

&Lt; Test Example 5: Analysis of protein expression level associated with itching >

To investigate the expression pattern of itch - related MDCs in HaCaT cells induced by inflammation, addition of P. yezoenisis extracts at various concentrations (40, 200, 1000 ㎍ / ㎖).

The ELISA test was used for the quantification of the MDC protein, and the procedure was carried out according to the method of ELISA kit (R & D systems, USA).

FIG. 3A shows MDC protein expression levels when HaCaT cells induced inflammation with IFNγ, and FIG. 3B shows the results when HaCaT cells induced inflammation with TNF? MDC &lt; / RTI &gt; protein expression patterns.

As shown in FIGS. 3A and 3B, in the case of treating HaCaT cells treated with IFN-y to induce inflammation, 40, 200, and 1000 μg / ml of ethanol extract of Kimchi significantly reduced MDC protein production Respectively.

That is, the ethanol extract of Kimchi of the present invention showed a slight decrease in the expression level of MDC but a marked decrease in the protein production. Therefore, the composition according to the present invention is effective in suppressing the itch-related MDC, TARC gene and protein expression, and reducing ET-1, PAR-2 gene and protein expression associated with itching of the skin, Prevention or treatment of the disease.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the same is by way of illustration and example only and is not to be construed as limiting the scope of the present invention. Accordingly, the actual scope of the present invention will be defined by the appended claims and their equivalents.

Claims (19)

A pharmaceutical composition for preventing or treating itching comprising an extract of Kim as an active ingredient. The composition according to claim 1, wherein the extract is water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. The composition according to claim 2, wherein the mixed solvent is 20 to 80% by weight of an aqueous solution of methanol, ethanol, butanol or propanol. The composition according to claim 2, wherein the mixed solvent is 20 to 80% by weight aqueous ethanol solution. 2. The composition of claim 1, wherein the itch is induced by the expression of MDC, TARC, ET-1, PAR-2. The method according to claim 1, wherein the itching is selected from the group consisting of seizure itch, winter itch, anal itch, vulvar itch, scrotum itch, watery itch, scalp itch, nasal itch, itchy mucus, itchy mucus and itchy eyes &Lt; / RTI &gt; The method according to claim 1, wherein the itching is associated with an internal medical disorder selected from the group consisting of biliary itch, chronic renal failure, malignancy, iron deficiency anemia, intracellular hypertension, hyperthyroidism, hypothyroidism, diabetes and acquired immunodeficiency Lt; RTI ID = 0.0 &gt; itch. &Lt; / RTI &gt; [2] The method of claim 1, wherein the itching is an itch accompanied by mental skin disease selected from the group consisting of chronic simple poisons, itching rash, hairy wall, nervous scraping wound, skin- / RTI &gt; A food composition for improving or preventing itching comprising an extract of Kim as an active ingredient. The composition according to claim 9, wherein the extract is water, a lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. The composition according to claim 10, wherein the mixed solvent is 20 to 80% by weight of an aqueous solution of methanol, ethanol, butanol or propanol. The composition of claim 10, wherein the mixed solvent is 20 to 80% by weight aqueous ethanol solution. 10. The composition of claim 9, wherein the itch is induced by the expression of MDC, TARC, ET-1, PAR-2. 10. The method according to claim 9, wherein the itching is selected from the group consisting of paroxysmal itch, winter itch, anal itching, scaly itch, scrotum itch, watery itch, scalp itch, nasal itch, itchy mucus, &Lt; / RTI &gt; 10. The method of claim 9, wherein the itching is associated with a medical condition selected from the group consisting of: itching of bile, chronic renal failure, malignancy, iron deficiency anemia, intracellular hypertension, hyperthyroidism, hypothyroidism, diabetes and acquired immunodeficiency Lt; RTI ID = 0.0 &gt; itch. &Lt; / RTI &gt; 10. The method according to claim 9, wherein the itching is characterized by itching accompanied by a mental skin disease selected from the group consisting of chronic simplex poisoning, itchy rash, hairy wall, nervous scraping wound, skin- / RTI &gt; 10. The composition of claim 9, wherein the food composition is selected from the group consisting of capsules, tablets, powders, granules, liquids, pills, flakes, pastes, syrups, gels, jellies and jars. A cosmetic composition for improving or preventing itching comprising an extract of Kim as an active ingredient. The cosmetic composition according to claim 7, wherein the formulation of the cosmetic composition is in the form of a tonic, a shampoo, a rinse, a hair conditioner, a hair spray, a powder, a gel, a cream, an essence, a lotion, a sol gel, an emulsion, an oil, a wax, / RTI &gt;

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