KR20140128123A - Composition for prevention and treatment of atopic dermatitis and inflammatory disorders comprising extract of Carpesium abrotanoides - Google Patents

Composition for prevention and treatment of atopic dermatitis and inflammatory disorders comprising extract of Carpesium abrotanoides Download PDF

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KR20140128123A
KR20140128123A KR20130046899A KR20130046899A KR20140128123A KR 20140128123 A KR20140128123 A KR 20140128123A KR 20130046899 A KR20130046899 A KR 20130046899A KR 20130046899 A KR20130046899 A KR 20130046899A KR 20140128123 A KR20140128123 A KR 20140128123A
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South Korea
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extract
tobacco
weight
present
inflammatory diseases
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KR20130046899A
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Korean (ko)
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김재화
오세량
지다정
송혁환
강호범
김두영
류형원
안경섭
이형규
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한국생명공학연구원
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Publication of KR20140128123A publication Critical patent/KR20140128123A/en

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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783—Angiosperms [Magnoliophyta]
    • A61K8/9789—Magnoliopsida [dicotyledons]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH

Abstract

The present invention relates to a pharmaceutical composition containing an extract of Carpesium abrotanoides as an active ingredient for preventing and treating atopic dermatitis and inflammatory disorders. More specifically, the extract of Carpesium abrotanoides in the present invention significantly decreases the expression of inflammatory cytokines including interferon gamma (IFNγ) and IL-10 at the transcriptional level, and in comparison with dexamethasone (DEX), the conventional anti-inflammatory drug, the DEX only inhibits the expression of IFNγ while the extract of Carpesium abrotanoides significantly inhibits the expression of both IFNγ and IL-10, thereby being able to be useful as a pharmaceutical composition, a skin external preparation composition, a cosmetic composition, and a healthy food composition for preventing and treating atopic dermatitis and inflammatory disorders.

Description

TECHNICAL FIELD The present invention relates to a pharmaceutical composition for prevention and treatment of atopic and inflammatory diseases containing an extract of tobacco grass as an active ingredient,

The present invention relates to a tobacco plant, The present invention relates to a pharmaceutical composition for preventing and treating atopic and inflammatory diseases,

Activation of NF-κB is regulated at various levels by positive and negative regulatory elements. In resting state, the NF-κB dimer exists in association with IκB for its inhibition. When IκB is phosphorylated by the IκB kinase (IκB kinase) complex, ubiquitination and proteosome degradation proteosomal degradation. Thus, NF-κB migrates to the nucleus, and the migrated NF-κB binds to the NF-κB by immunity, cell differentiation, proliferation, apoptosis, survival response and stress response stress response). In addition, the activity of NF-κB is involved not only in cancer, but also in inflammatory diseases, autoimmune diseases and metabolic diseases.

IFNγ and IL-10 are well-known genes that are induced by NF-κB activity. IFNγ is mainly produced by activated T cells and natural killer cells, which are responsible for innate immunity and adaptive immunity to the intracellular pathogen, and tumor-regulating and immune-stimulatory ) And immune-modulatory effects. IL-10 is a cytokine with anti-inflammatory properties and has a great influence on the development of the immune response. T helper cell tissues composed of TH 1 or TH 2 cells generally produce IL-12, IFNγ and TNF when exposed to IFNγ producing IL-10.

Glucocorticoids are inflammatory diseases such as asthma, rheumatoid arthritis, allergic disease and autoimmune disease associated with overexpression of an inflammatory gene. ≪ / RTI > is generally used for the inhibition of inflammation in various types of inflammatory diseases. Glucocorticoids affect reactive cells due to the activity of the glucocorticoid receptor (GR) in order to directly or indirectly regulate transcriptional regulation of the target gene. GR may be associated with NF-κB by a mechanism similar to direct protein-protein interaction by inhibiting the expression of a range of inflammatory genes.

Psychological problems cause a decrease in natural killer cell (NK cell) activity and non-normal regulation of cytokine stability. NK cell activity responds to the impact of psychological stress, respectively. Psychological distress alters not only the production of interferon gamma (IFN-γ) but also the mode of cytokine production. Glucocorticoid not only reduces the sustained production of cytokines, IL-6, TNF [alpha] and IFN-y, but also reduces the ability of NK cells to bind to tumor targets. However, glucocorticoids do not reduce cytokines, IL-10.

Traditionally, natural products have been used for the treatment of inflammation and other diseases. Aspirin is the most famous natural-derived drug that is commonly used as an anti-inflammatory agent. Other essential anti-inflammatory agents used in asthma control include corticosteroids and salbutamol. In addition, many other natural products such as cyclosporine, mizoribine and FK-506 have shown to inhibit inflammation.

Tobacco Pool ( Carpesium abrotanoides ) are plants belonging to the Campanulales Asteraceae , which grow in the bushes at the edge of forests and are distributed in Korea, China and Japan. The tobacco pool is well known as a "barefoot doctor" and is known to be used as a constitutional remedy, a scurvy remedy, a depurative, a discutient, an emetic, an expectorant, a febrigufe, (laxative) and wound treatment (vulnerary) are known to be effective. Tobacco juice is used to treat bronchitis, tonsillitis, boils, ulcers, and snake bites. The stem juice is used when bitten by insects, and the stem juice is also very effective in the treatment of sore throat.

Accordingly, the inventors of the present invention have been developing a therapeutic agent for inflammatory diseases derived from natural products that can overcome the side effects and limitations of the anti-inflammatory drugs used in the past, abrotanoides) extract the inflammatory cytokine (cytokine) interferon and significantly reduced the expression level of the gamma and IL-10, the conventional anti-inflammatory agent dexamethasone (dexamethasone; when compared with DEX), DEX is present a suppressed only expression of interferon gamma The tobacco grass extract of the present invention is useful as a pharmaceutical composition, an external preparation for skin, a cosmetic composition and a health food composition for preventing or treating atopic and inflammatory diseases by significantly inhibiting the expression of interferon gamma as well as IL-10 The present invention has been completed.

An object of the present invention is snuffbox (Carpesium The present invention also provides a pharmaceutical composition for the prevention and treatment of atopic and inflammatory diseases, which comprises as an active ingredient an abrotanoides extract.

The present invention relates to a tobacco plant, The present invention provides a pharmaceutical composition, an external preparation for skin, a cosmetic composition and a health food composition for the treatment of atopic and inflammatory diseases, which comprises as an active ingredient an abrotanoides extract.

The tobacco plant of the present invention ( Carpesium abrotanoides extract significantly reduced the expression of interferon gamma and IL-10, which are inflammatory cytokines, at the transcriptional level and inhibited only the expression of interferon gamma when compared to the conventional anti-inflammatory agent dexamethasone (DEX) The tobacco grass extract of the present invention is useful as a pharmaceutical composition for preventing or treating atopic and inflammatory diseases, a composition for external application for skin, a cosmetic composition and a health food composition by significantly inhibiting not only interferon gamma but also IL-10 expression Can be used.

Figure 1 is a schematic view of a tobacco plant abrotanoides < / RTI > extract inhibits the production of interferon gamma;
Carpesium: Tobacco grass extract,
Fraction 1: Tobacco grass fraction 1,
Fraction 2: Tobacco grass fraction 2, and
Fraction 3: Tobacco grass fraction 3.
Figure 2 shows that the tobacco grass extract inhibits the activity of NF-κB;
Carpesium: Tobacco grass extract.
FIG. 3 shows that tobacco grass extract inhibits mRNA expression of interferon gamma;
Carpesium: Tobacco grass extract, and
DEX: dexamethasone.

Hereinafter, the present invention will be described in detail.

Tobacco Pool ( Carpesium The present invention also provides a pharmaceutical composition for the prevention and treatment of atopic and inflammatory diseases, which comprises as an active ingredient an abrotanoides extract.

The tobacco grass extract and fractions are preferably, but not necessarily, prepared by a process comprising the steps of:

1) extracting with a tobacco extract solution;

2) filtering the extract of step 1);

3) concentrating the filtered extract of step 2) under reduced pressure and then drying to produce an extract of tobacco grass; And

4) Extracting the tobacco grass extract of step 3) with an organic solvent to prepare a tobacco grass fraction.

In the above method, the tobacco plant of step 1) may be used without limitation, such as cultivated or commercially available plants. The tobacco paste may be any of leaves, stems, roots, or flowers, but is not limited thereto.

In the above method, it is preferable to use water, an alcohol or a mixture thereof in the extraction solvent of the step 1). As the alcohol, C 1 to C 2 lower alcohol is preferably used, and as the lower alcohol, ethanol or methanol is preferably used. As the extraction method, it is preferable to use shaking extraction, Soxhlet extraction or reflux extraction, but it is not limited thereto. It is preferable that the extraction solvent is added by 1 to 10 times the amount of the dried tobacco paste, more preferably by 2 to 3 times. The extraction temperature is preferably 20 占 폚 to 100 占 폚, more preferably 20 占 폚 to 40 占 폚, and most preferably room temperature, but is not limited thereto. The extraction time is preferably 10 to 48 hours, more preferably 15 to 30 hours, most preferably 24 hours, but is not limited thereto. The number of times of extraction is preferably 1 to 5 times, more preferably 3 to 4 times, and most preferably, 3 times, but is not limited thereto.

In the above method, it is preferable to use a vacuum decompression concentrator or a vacuum rotary evaporator for the decompression concentration in step 3), but it is not limited thereto. The drying is preferably performed under reduced pressure, vacuum drying, boiling, spray drying or freeze drying, but not always limited thereto.

In the above method, the organic solvent in step 4) is preferably n-hexane, chloroform, ethyl acetate or butanol, but is not limited thereto. The fraction may be n-hexane fraction, chloroform fraction, ethyl acetate fraction, butanol fraction or water fraction obtained by suspending the tobacco grass extract in water and sequentially fractionating the fraction with n-hexane, chloroform, ethyl acetate, butanol and water But is not limited thereto. The fraction may be obtained by repeating the fractionation process from the tobacco extract at 1 to 5 times, preferably 3 times, followed by fractionation and concentration under reduced pressure. However, the fraction is not limited thereto.

The inflammatory diseases include inflammatory diseases such as dermatitis, conjunctivitis, periodontitis, rhinitis, otitis, sore throat, tonsillitis, pneumonia, gastric ulcer, gastritis, Crohn's disease, colitis, hemorrhoids, gout, ankylosing spondylitis, rheumatic fever, lupus, fibromyalgia, , Rheumatoid arthritis, shoulder inflammation, tendinitis, hay fever, perianal inflammation, myositis, hepatitis, cystitis, nephritis, sjogren's syndrome, multiple sclerosis and acute and chronic inflammatory diseases But is not limited thereto.

In a specific embodiment of the present invention, the present inventors prepared a tobacco grass extract using a tobacco plant, treated the extracted tobacco grass extract with NK92 cells under various conditions, and then measured the amount of interferon gamma produced using an ELISA kit , It was confirmed that the tobacco grass extract decreased the production of interferon gamma in a concentration-dependent manner (see FIG. 1).

In addition, the present inventors conducted a promoter analysis to confirm whether the result was due to the inhibitory effect of the NF-κB transcription factor on tobacco grass. As a result, the tobacco grass extract inhibited the activity of NF-κB transcription factor Inhibit the production of interferon gamma (see FIG. 2).

In addition, the present inventors confirmed mRNA expression levels to confirm that interferon gamma production inhibition occurs at the transcription level. As a result, it was confirmed that the tobacco grass extract significantly reduced the expression of IFNγ and IL-10 (see FIG. 3).

Accordingly, it has been confirmed that the tobacco grass extract of the present invention significantly inhibits the production of both interferon gamma and IL-10, and thus can be effectively used as a pharmaceutical composition for the treatment and prevention of atopic and inflammatory diseases.

The composition containing the tobacco grass extract of the present invention may further contain at least one active ingredient which exhibits the same or similar function in addition to the above components.

The composition of the present invention may further comprise a pharmaceutically acceptable additive, wherein pharmaceutically acceptable additives include starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, calcium hydrogen phosphate, lactose Starch glycolate, sodium starch glycolate, carnauba wax, synthetic aluminum silicate, stearic acid, magnesium stearate, aluminum stearate, calcium stearate, calcium stearate, , White sugar, dextrose, sorbitol and talc. The pharmaceutically acceptable additives according to the present invention are preferably included in the composition in an amount of 0.1 to 90 parts by weight, but are not limited thereto.

That is, the composition of the present invention can be administered in various formulations of oral and parenteral administration at the time of actual clinical administration. In the case of formulation, a diluent such as a filler, an extender, a binder, a wetting agent, a disintegrant, . ≪ / RTI > Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose, Sucrose, lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate talc may also be used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions and syrups, and various excipients such as wetting agents, sweetening agents, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are simple diluents commonly used . Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used as the non-aqueous solvent and suspension agent. Examples of suppository bases include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, and the like.

The composition of the present invention may be administered orally or parenterally in accordance with the intended method, and may be administered orally, parenterally or intraperitoneally, rectally, subcutaneously, intravenously, intramuscularly, . The dosage varies depending on the patient's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and severity of disease.

The composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level will depend on the type of disease, severity, , Sensitivity to the drug, time of administration, route of administration and rate of release, duration of treatment, factors including co-administered drugs, and other factors well known in the medical arts. The composition of the present invention can be administered as an individual therapeutic agent or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents, and can be administered singly or in multiple doses. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without side effects, which can be easily determined by those skilled in the art.

Specifically, the effective amount of the compound according to the present invention may vary depending on the age, sex, and body weight of the patient. In general, 0.1 mg to 100 mg, preferably 0.5 mg to 10 mg per kg of body weight is administered daily or every other day Or one to three times a day. However, the dosage may be varied depending on the route of administration, the severity of obesity, sex, weight, age, etc. Therefore, the dosage is not limited to the scope of the present invention by any means.

In addition, the present invention provides an external preparation for skin and cosmetic composition for prevention and improvement of atopic and inflammatory diseases containing tobacco grass extract as an active ingredient.

The inventors of the present invention confirmed that the tobacco grass extract of the present invention significantly inhibited the production of both interferon gamma and IL-10, and thus could be effectively used as an external preparation for skin and cosmetic composition for prevention and improvement of atopic and inflammatory diseases .

In addition, the tobacco grass extract of the present invention is preferably used as a raw material for cosmetics, bath products, soaps, pharmaceutical ointments, animal pharmaceuticals or pack products.

That is, the therapeutic agent for atopic and inflammatory diseases containing the tobacco grass extract of the present invention as an active ingredient can be prepared as a solution, a suspension, a spray, a patch, a pad, a cream, an ointment, a gel, a tablet or a pill.

The solution may contain, in addition to the active compound, conventional excipients such as solvents, solubilizers and emulsifiers such as water, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, Glycerin, natural corn stover oil, olive oil, castor oil, almond oil and sesame oil, glycerol, glycerol formal alcohol, tetramethyluronium hexafluorophosphate, Fatty acid esters of sorbitan, furfuryl alcohol, polyethylene glycol and sorbitan, or mixtures of these substances, and may contain methyl or propyl-p-hydroxybenzoate or sorbic acid as a preservative.

In addition to the active compound, the suspending agent may contain conventional excipients such as liquid diluents (e.g., water, ethyl alcohol and propylene glycol, polyethylene glycol), and suspending agents (e.g., ethoxylated isostearyl alcohol, polyoxyethylene sorbitol, sorbitan Esters, cellulose derivatives and preservatives for hydrogenation), microcrystalline cellulose, aluminum metahydroxide, bentonite, agar and tragacanth, injectable esters such as ethyl oleate, or mixtures of these substances.

The ointments, creams and gels may contain conventional excipients such as animal and vegetable fats, wax paraffin, starch, targacans, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, ≪ / RTI >

In addition, the administration method can be generally applied to the affected part, and the preferable administration method can be selected according to the formulation.

The daily dose of the composition of the present invention is determined depending on the subject to be administered, method of administration, and symptom, but generally, the tobacco grass extract contained in the composition and the carabrone or its pharmaceutically acceptable salt represented by the formula (1) 100 < RTI ID = 0.0 > pg / ml. ≪ / RTI > The number of times of administration is preferably two times a day or more, but the number of times of administration can also be adjusted depending on the symptom level. However, the dosage needs to be varied and can be varied, in particular considering the sperm specificity and weight of the object to be treated, the type and depth of the disease, the nature of the formulation, the nature of the drug administration, and the duration or interval of administration.

In order to effectively treat atopic and inflammatory diseases, it is important to select a suitable external preparation. In order to treat atopic and inflammatory diseases, tobacco grass extract may be suitably diluted and applied directly to the skin. In the present invention, an ointment agent capable of effectively applying the atopic and inflammatory therapeutic agents to the affected part is provided.

The ointment of the present invention is prepared by combining the above-described therapeutic agent for atopic and inflammatory diseases with an inorganic substance and then coating it with a liposoluble base. The inorganic material is preferably a material having excellent antimicrobial activity, anti-inflammatory effect, epidermal regeneration effect, etc. Specific examples thereof include zinc oxide, zinc carbonate, iron oxide and the like.

In addition, it is preferable to further use a ceramic carrier capable of safely impregnating the atopy of the present invention, which is a water-soluble substance, with a therapeutic agent for inflammatory diseases. As the ceramic carrier, zeolite, talc, gypsum, mortar and mixtures thereof are preferably used. Such a ceramic carrier is excellent in the impregnation property of the water-soluble component, so that the water-soluble component can be smoothly supplied to the skin.

In addition, the present invention provides a health food composition for preventing and ameliorating atopic and inflammatory diseases containing an extract of tobacco grass as an active ingredient.

The inventors of the present invention have confirmed that the tobacco grass extract of the present invention can be effectively used as a health food composition for prevention and improvement of atopic and inflammatory diseases by significantly inhibiting the production of both interferon gamma and IL-10.

The health food of the present invention can be used as it is, or can be used in combination with other food or food ingredients, and can be suitably used according to conventional methods.

There is no particular limitation on the kind of the health food. Examples of the foods to which the tobacco grass extract can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, ice cream, soups, Alcoholic beverages, and vitamin complexes, all of which include health foods in a conventional sense.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. Such natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like. The ratio of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 of the composition of the present invention.

In addition to the above, the health food of the present invention may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acids and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, , A carbonating agent used in carbonated drinks, and the like. It may also contain flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The proportion of such additives is not critical, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

Hereinafter, the present invention will be described in detail with reference to Examples and Production Examples.

However, the following examples and preparations are merely illustrative of the present invention, and the content of the present invention is not limited by the following examples and production examples.

< Example  1> Tobacco Pool ( Carpesium abrotanoides ) Preparation of extract

Tobacco grass extracts were obtained by pulverizing 200 g of tobacco leaves obtained from China with a pulverizer and then extracting 700 ml of an aqueous methanol solution (80% of methanol and 20% of water). The extract solution was filtered using a filter paper, and the filtered extract was concentrated with a vacuum concentrator to obtain 10 g of tobacco grass extract.

< Example  2> Interferon gamma of tobacco grass extract IFN γ) production inhibitory effect

<2-1> Cell culture

The cultivation of human cell lines (NK 92, NKys, U937) was carried out at 37 ° C in a humidified atmosphere of 5% CO 2 . The blood cell lines NK 92, NKys (human NK cell line) and U937 were purchased from the American Type Culture Collection (ATCC), and the cell lines were cultured in DMEM supplemented with 10% FCS (HyClone, Logan, UT), 2 mM L- glutamate, (Life Technologies, Karlsruhe, Germany) medium containing 100 μg / ml penicillin, 100 μg / ml streptomycin (Life Technologies, Karlsruhe, Germany)

<2-2> Confirmation of inhibitory effect of tobacco grass extract on interferon gamma production

In order to confirm inhibition of interferon gamma production of the tobacco grass extract prepared in Example 1, tobacco grass extract was treated with NK92 cells under various conditions, and the amount of interferon gamma produced was measured using an ELISA kit (BD Biosciences) Respectively.

Specifically, tobacco grass extract was treated with NK92 cell line under various conditions, and the cell line was cultured. Then, 50 μl of the cell culture supernatant was added to an assay plate coated with an antibody recognizing IFNγ, Lt; / RTI &gt; After a period of reaction, an antibody recognizing IFNγ bound to HRP is added and cultured for an additional hour, and the unreacted mixture is washed. The bound enzyme is visualized by the addition of a TMB component to the substrate and the reaction can be stopped by adding 100 μl of 2N H 2 SO 4 . The absorbance was also measured using an ELISA reader (Molecular Devices E max) at 450 nm.

As a result, as shown in Fig. 1, it was confirmed that the tobacco grass extract decreased the production of interferon gamma produced in the activated blood cell NK92 cell line in a concentration-dependent manner (Fig. 1).

< Example  3> Tobacco grass extract NF Inhibitory Effect of -KB Transcription Factor

As a result of the above Example <2-2>, a promoter analysis was performed to determine whether tobacco grass extract was due to the inhibitory effect of NF-κB transcription factor.

Specifically, 1 × 10 5 cells / ml HEK293 cell line was prepared and 1 μg of pGL4.32 [luc2P / NF-κB-RE / Hygro] vector was transfected with a DNA transfection reagent KINOVATE). The transfected cell line was stimulated with 500 ng / ml PMA and serial diluted tobacco grass extract from 10 / / ㎖ to 78 ng / ㎖ was added and cultured for 6 hours. Then, luciferase cell lysis buffer (Promega) was added and stirred at 4 캜 for 30 minutes to obtain a lysate of the stimulated cells. 10 μl of the obtained lysate was transferred to a 96-well luminometer-plate, and 50 μl of Luciferase Assay Substrate (Promega) was added thereto. Lumat Plus LB).

As a result, as shown in FIG. 2, it was confirmed that tobacco grass extract inhibited the production of interferon gamma through inhibition of NF-κB transcription factor activity (FIG. 2). Generally, the production of interferon gamma in immune cells depends on the activity of NF-κB, and the ability of NF-κB to inhibit activity inhibits the activity of pro-inflammatory cytokines.

< Example  4> Interferon gamma of tobacco grass extract and IL -10 mRNA  Confirming expression suppression effect

To confirm that the inhibition of interferon gamma production in Example <2-2> occurs at the transcription level, RT-PCR was performed on the NK92 cell line to measure the transcriptional activity of interferon gamma and IL-10.

Specifically, 5 × 10 5 cells / ml of NK92 cells cultured in a 6-well plate containing IL-2 as a growth factor was treated with 500 ng / ml PMA, and then treated with 5, 1, 0.1 μg / ml Of tobacco grass extract was added. After 15 hours of incubation, the stimulated cell lines were collected and pellets were obtained using a micro-centrifuge. A TRIzol solution (MRC) was added to the obtained pellet to isolate the total RNA. A part of the separated total RNA was dried and partially dissolved in 50 μl of diethylene-pyrocarbonate (DEPC). The purified RNA was confirmed to have a high purity using a UV spectrophotometer (Nano Vue). Reverse transcription for synthesis of cDNA was carried out using the first strand cDNA synthesis kit (Promega) reaction was performed according to the manufacturer's instructions using the AccuScript High Accuracy First Strand cDNA Synthesis Kit (AccuScript High Fidelity 1st Strand cDNA Synthesis Kit, Stratagene).

PCR was performed to amplify the synthesized cDNA, and 1 μl of the synthesized cDNA was added to 20 μl of a PCR mixture composed of 0.5 U ExTaq DNA polymerase, 1 × buffer and 1 mM dNTP mix (Takara) and specific primer pairs And the PCR reaction was carried out. The following primers used in the PCR were designed using the Primer 3 program and purchased from Bioneer (Daejeon, Korea);

GAPDH_sense (SEQ ID NO: 1): 5'-CCATCACCATCTTCCAGGAG-3 '

GAPDH_antisense (SEQ ID NO: 2): 5'-ACAGTCTTCTGGGTGGCAGT-3 '

IFNgamma_sense (SEQ ID NO: 3): 5'-TGGCTGAACTGTCGCCAGCA-3 '

IFNgamma_antisense (SEQ ID NO: 4): 5'-TGGCTGCCTAGTTGGCCCCT-3 '

IL-10_sense (SEQ ID NO: 5): 5'-TGCCTTCAGCAGAGTGAAGA-3 ', and

IL-10_antisense (SEQ ID NO: 6): 5'-GCCACCCTGATGTCTCAGTT-3 '.

Amplification by PCR reaction was carried out using GeneAmp PCR system 2700 (Applied Biosystems, USA) under the following conditions; After repeating 5 to 9 minutes at 94 ° C, 45 seconds at 94 ° C, 45 seconds at 56 ° C and 25 to 40 times at 72 ° C for 1 minute, the final extension reaction was carried out at 72 ° C for 7 minutes. The PCR products were separated on 1.5% agarose gel, stained with EtBr (ethidium bromide), visualized with a Gel Doc 2000 UV trans-illuminator (UV trans-illuminator, Bio-Rad Laboratories, (Bio-Rad Laboratories, USA). Each sample was tested more than 3 times and representative data were presented.

As a result, as shown in FIG. 3, the expression of IFNγ and IL-10 was significantly reduced when the tobacco grass extract was treated with NK92 cell line, a blood cell. In comparison with DEX, a glucocorticoid, which is used as an inhibitor of NF-κB used as a positive control, and tobacco grass extract of the present invention, similar effects were shown in the inhibition of IFNγ, but glucocorticoids While the tobacco grass extract of the present invention significantly inhibited IL-10 (FIG. 3).

< Manufacturing example  1> Preparation of pharmaceutical preparations

<1-1> Sanje  Produce

2 g of the tobacco grass extract of the present invention

Lactose 1 g

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

<1-2> Preparation of tablets

100 mg of the tobacco grass extract of the present invention

Corn starch 100 mg

100 mg of milk

2 mg of magnesium stearate

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

&Lt; 1-3 > Preparation of capsules

100 mg of the tobacco grass extract of the present invention

Corn starch 100 mg

100 mg of milk

2 mg of magnesium stearate

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

&Lt; 1-4 >

1 g of the tobacco grass extract of the present invention

Lactose 1.5 g

Glycerin 1 g

0.5 g of xylitol

After mixing the above components, they were prepared so as to be 4 g per one ring according to a conventional method.

<1-5> Preparation of granules

150 mg of the tobacco grass extract of the present invention

Soybean extract 50 mg

200 mg of glucose

600 mg of starch

After mixing the above components, 100 mg of 30% ethanol was added and the mixture was dried at 60 캜 to form granules, which were then filled in a capsule.

< Manufacturing example  2> Manufacture of ointment preparation

<2-1> Preparation of liposoluble (W / O formulation) ointment

The tobacco grass extract of the present invention is 0.01 to 10.0%

Stearyl alcohol 7.0%

Stearic acid 2.2%

Squalene 3.4%

Octyl dodecanol 5.0%

Olive oil 6.0%

Glycerin monoauric fatty acid ester 10.0%

Isopropyl adipate 2.0%

Tego 0.3%

Carboxyl vinyl polymer 3.0%

Propylene glycol 5.0%

Kojic acid 0.05%

Alpha-keto glutamic acid 0.2%

Cysteine 0.2%

Parabens 0.3%

Sterile purified water or 0.02 M phosphoric acid or intermediate solution

At the preparation of ointment, the components other than carboxyvinyl polymer and kojic acid were emulsified at 60 ° C or higher in the above composition components, and then carboxyvinyl polymer was added while stirring. After cooling, the remaining aqueous phase components such as kojic acid were slowly added to the mixture to homogeneity when they had a suitable viscosity, and then allowed to stand for maturation before being made into a finished product.

<2-2> Preparation of hydrophilic (W / O type) ointment

The tobacco grass extract of the present invention is 0.01 to 10.0%

White vaseline 25.0%

Stearyl alcohol 20.0%

Propylene glycol 12.0%

Propylene misstatin 0.1%

Sodium glycerin Mono Laurel 0.2%

Polyoxyethylene hardened castor oil 4.0%

Betaine-containing cationic emulsifier 0.2%

Glycerin monostearate 0.1%

0.1% paraoxybenzoic acid propylate

Purified water balance

An ointment type (O / W) ointment agent was prepared by the ointment preparation method of Example <2-1> with the above composition.

<2-3> Preparation of cream

The tobacco grass extract of the present invention is 0.01 to 10.0%

Liquid paraffin 6.0%

Squalane 4.0%

0.5% sorbitan sesquioleate

Polysorbate 60 1.5%

Beeswax 10.0%

Cappolic / capric triglyceride 5.0%

Butylene glycol 6.0%

Free ethanolamine 3.0%

Preservatives 2.0%

Fragrance

Purified water balance

A cream containing the tobacco grass extract in the above composition was prepared according to a conventional method.

< Manufacturing example  3> Cosmetics  Preparation of composition

<3-1> Production of cream

0.1 part by weight of tobacco grass extract of the present invention

Cetostearyl alcohol 2.8 parts by weight

2.6 parts by weight

Stearic acid 1.4 parts by weight

Glycerin monostearate 2 parts by weight

&Lt; tb &gt; &lt; tb &gt;

Sorbitol sesquioleate 1.4 parts by weight

Jojoba oil 4 parts by weight

Squalane 3.8 parts by weight

Polysorbate 60 1.1 parts by weight

Macadia oil 2 parts by weight

Tocopheryl acetate 0.2 part by weight

Methylpolysiloxane 0.4 part by weight

Ethylparaben 0.1 part by weight

Propylparaben 0.1 part by weight

Euxyl K-400 0.1 part by weight

1,3-butylene glycol 7 parts by weight

Methylparaben 0.05 part by weight

Glycerin 6 parts by weight

d-Pandenol 0.2 part by weight

Triethanolamine 0.2 part by weight

pt 41891 0.2 part by weight

Purified water 50.55 parts by weight

<3-2> Production of Lotion

0.1 part by weight of tobacco grass extract of the present invention

Cetostearyl alcohol 1.6 parts by weight

Stearic acid 1.4 parts by weight

Glycerin monostearate of pro-type 1.8 parts by weight

&Lt; tb &gt; &lt; tb &gt;

Sorbitol sesquioleate 0.6 parts by weight

Squalene 4.8 parts by weight

Macadia oil 2 parts by weight

Jojoba oil 2 parts by weight

Tocopheryl acetate 0.4 part by weight

0.2 part by weight of methylpolysiloxane

Ethylparaben 0.1 part by weight

Propylparaben 0.1 part by weight

4 parts by weight of 1,3-butylene glycol

0.1 part by weight of methylparaben

Xanthan gum 0.1 part by weight

Glycerin 4 parts by weight

d-Pandenol 0.15 part by weight

Allantoin 0.1 part by weight

Cargar (2% aq. Sol) 4 parts by weight

Triethanolamine 0.15 part by weight

Ethanol 3 parts by weight

pt 41891 0.1 part by weight

Purified water 51.7 parts by weight

< Manufacturing example  4> Cosmetics  Preparation of cleaning agents in compositions

<4-1> Production of soap

4.0 parts by weight of tobacco grass extract of the present invention

Soap base 94.25 parts by weight

Tocopherol acetate 0.3 part by weight

Glycerin 0.2 part by weight

PEG 4000 (polyethylene glycol 4000) 0.8 parts by weight

Jojoba oil 0.3 part by weight

0.15 parts by weight of cetanol

The components were mixed well in a mixer, extruded in a soap making machine, and cut and molded into a predetermined size to prepare a soap composition. The soap base may be a sodium salt of palm oil, which is a natural oil commonly used in the art.

<4-2> body Cleanser's  Produce

4.0 parts by weight of tobacco grass extract of the present invention

Ammonium lauryl sulfate 42 parts by weight

Cocamidopropyl betaine 12 parts by weight

Glycol distearate 0.5 part by weight

Propylene glycol 1.0 part by weight

Pigment amount

Fragrance appropriate amount

Citric acid suitable amount

Drain (50% concentrated liquid) 1.0 part by weight

Purified water is added to make 100 parts by weight.

Ammonium lauryl sulfate, cocamidopropyl betaine, glycol distearate and propylene glycol were heated to 70 ° C and emulsified with heated purified water, and the remaining components were added and cooled to room temperature to prepare a body cleanser.

< Manufacturing example  6> Manufacture of health food

<6-1> Manufacture of Flour Food

0.5 to 5.0 parts by weight of the tobacco grass extract of the present invention was added to wheat flour, and the mixture was used to prepare bread, cake, cookies, crackers and noodles.

<6-2> soup  And juicy ( gravies )

0.1 to 5.0 parts by weight of the tobacco grass extract of the present invention were added to soups and juices to prepare health promotion meat products, noodle soups and juices.

<6-3> Ground Beef  Produce

10 parts by weight of the tobacco grass extract of the present invention was added to ground beef to prepare ground beef for health promotion.

<6-4> Dairy products ( dairy products )

5-10 parts by weight of the tobacco grass extract of the present invention was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

<6-5> Solar  Produce

Brown rice, barley, glutinous rice, and yulmu were dried by a known method and dried, and the mixture was granulated to a powder having a particle size of 60 mesh.

Black soybeans, black sesame seeds, and perilla seeds were steamed and dried by a conventional method, and then they were prepared into powder having a particle size of 60 mesh by a pulverizer.

The tobacco grass extract of the present invention was concentrated under reduced pressure in a vacuum concentrator, spray dried, and dried with a hot air drier, and the resulting dried product was pulverized to a size of 60 mesh with a pulverizer to obtain a dry powder.

The grains, seeds and the tobacco grass extract of the present invention prepared above were blended in the following proportions.

(30 parts by weight of brown rice, 15 parts by weight of yulmu, 20 parts by weight of barley)

Seeds (7 parts by weight of perilla, 8 parts by weight of black beans, 7 parts by weight of black sesame seeds)

Tobacco grass extract (3 parts by weight) of the present invention,

(0.5 part by weight),

(0.5 parts by weight)

< Manufacturing example  7> Manufacturing of beverages

<7-1> Health drink  Produce

(5 g) of the tobacco grass extract of the present invention was uniformly blended with a sub ingredient such as liquid fructose (0.5%), oligosaccharide (2%), sugar (2%), salt (0.5% And then packaged in small containers such as glass bottles and plastic bottles.

<7-2> Manufacture of vegetable juice

5 g of the tobacco grass extract of the present invention was added to 1,000 ml of tomato or carrot juice to prepare vegetable juice.

<7-3> Production of fruit juice

1 g of the tobacco grass extract of the present invention was added to 1,000 ml of apple or grape juice to prepare fruit juice.

<110> Korea Research Institute of Bioscience and Biotechnology <120> Method for infinite reproduction of somatic cells for inducing          induced pluripotent stem cells by formation of teratoma <130> 13P-02-20 <160> 16 <170> Kopatentin 1.71 <210> 1 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> Sox2-F <400> 1 ccatgtatag atctggagga aa 22 <210> 2 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> Sox2-R <400> 2 caaatattaa aacttttttt tcatcg 26 <210> 3 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Oct4-F <400> 3 cccaggtccc cactttggca c 21 <210> 4 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> Oct4-R <400> 4 acccctgttg tgcttttaat ccc 23 <210> 5 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> Nanog-F <400> 5 caccagtgga gtatcccagc at 22 <210> 6 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> Nanog-R <400> 6 tggcagagaa gttttgctgc aac 23 <210> 7 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Zfp42-F <400> 7 catcctaacc cacgcaaagg 20 <210> 8 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Zfp42-R <400> 8 aagagttgca aaattatttt 20 <210> 9 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> FSP-F <400> 9 aggccctgga tgtaattgtg 20 <210> 10 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> FSP-R <400> 10 gctgtccaag ttgctcatca 20 <210> 11 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Fap-F <400> 11 tcaactgtga tggcaagagc 20 <210> 12 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Fap-R <400> 12 catggttctg gtcagagtac 20 <210> 13 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Fap-1F <400> 13 tcaactgtga tggcaagagc 20 <210> 14 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Fap-1R <400> 14 gtaccacatc gcctggaaat 20 <210> 15 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> GAPDH-F <400> 15 ccagtatgac tccactcacg 20 <210> 16 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> GAPDH-R <400> 16 ttcacaccca tcacaaacat 20

Claims (8)

Tobacco Pool ( Carpesium The present invention relates to a pharmaceutical composition for the prevention and treatment of atopic and inflammatory diseases, which comprises as an active ingredient an abrotanoides extract.
The pharmaceutical composition according to claim 1, wherein the tobacco extract is extracted from leaves, stems or flowers of a tobacco plant.
The pharmaceutical composition according to claim 1, wherein the extract is extracted with water, a C 1 to C 2 lower alcohol or a mixture thereof as a solvent.
4. The pharmaceutical composition according to claim 3, wherein the lower alcohol is ethanol or methanol.
The method of claim 1, wherein the inflammatory disease is selected from the group consisting of dermatitis, conjunctivitis, periodontitis, rhinitis, otitis, sore throat, tonsillitis, pneumonia, gastric ulcer, gastritis, Crohn's disease, colitis, hemorrhoids, gout, ankylosing spondylitis, rheumatic fever, lupus, fibromyalgia ), Rheumatoid arthritis, osteoarthritis, rheumatoid arthritis, periarthritis, tendinitis, hay fever, tendinitis, myositis, hepatitis, cystitis, nephritis, sjogren's syndrome, multiple sclerosis and acute and chronic inflammatory diseases Or a pharmaceutically acceptable salt thereof.
An external skin preparation for prevention and improvement of atopic and inflammatory diseases containing tobacco grass extract as an active ingredient.
A cosmetic composition for improving atopic and inflammatory diseases containing an extract of tobacco grass as an active ingredient.
A health food composition for preventing and ameliorating atopic and inflammatory diseases containing tobacco grass extract as an active ingredient.

KR20130046899A 2013-04-26 2013-04-26 Composition for prevention and treatment of atopic dermatitis and inflammatory disorders comprising extract of Carpesium abrotanoides KR20140128123A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20220067975A (en) * 2020-11-18 2022-05-25 한국과학기술연구원 Composition for anti-allergy comprising extract of carpesium cernuum

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR20220067975A (en) * 2020-11-18 2022-05-25 한국과학기술연구원 Composition for anti-allergy comprising extract of carpesium cernuum

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