KR101207239B1 - A composition for the prevention and treatment of inflammatory disease comprising the fractions of Asparagus cochinchinensis as an active ingredient - Google Patents

Abstract

The present invention is the Asparagus The present invention relates to a composition for the prevention and treatment of inflammatory diseases containing a fraction prepared from the extract of cochinchinensis as an active ingredient. More specifically, the fraction prepared by fractionating the astronomical extract with an organic solvent is an interleukin-1β (inflammatory cytokine). Since it is a non-cytotoxic safe substance that inhibits the production or secretion of interleukin-1β, IL-1β) and tumor necrosis factor-α (TNF-α), the composition for the prevention or treatment of inflammatory diseases It can be usefully used as an active ingredient.

Description

A composition for the prevention and treatment of inflammatory disease comprising the fractions of Asparagus cochinchinensis as an active ingredient}

The present invention relates to a composition for the prevention and treatment of inflammatory diseases containing a fraction prepared from the astronomical extract as an active ingredient.

Inflammation is one of tissue damage, an external stimulus, or a protective response of biological tissues to various infectious agents. Enzyme activation, inflammatory mediator secretion, cell infiltration due to organic interaction of various inflammatory mediators and various immune cells in blood vessels and body fluids And a series of complex pathologies, including fluid effusion, circulatory disorders, tissue degeneration and hyperproliferation. In the inflammatory process, macrophages initially gather into the wound and attack the invading bacteria, and then plasma builds up in the wound and increases blood flow, causing external symptoms such as fever, erythema, edema, and pain. . If these inflammatory reactions occur continuously or excessively, the disease progresses to the main pathology of the disease (sensitized allergic disease, chronic inflammatory disease) and causes serious abnormal disorders.

Various biochemical phenomena are involved as a cause of inflammation in living bodies. Macrophage (macrophage) is a cell with a variety of functions to produce various cytokines and nitric oxide (NO) by chemical stimulation plays an important role in the inflammatory response. Inducible nitric acidase, especially expressed by cytokine stimulation such as lipopolysaccharide (LPS), interferon-γ, and tumor necrosis factor-α (TNF-α) in macrophages oxide synthase (iNOS) produces large amounts of NO over long periods of time. This oxidative stress is known to promote NFκB activity, a transcription factor of the inflammatory response inhibited by IκB. Activated NFκB is known to migrate to the nucleus and stimulate the expression of genes that induce inflammatory responses, such as iNOS, COX-2 and several cytokines such as interleukin-1β and IL-1β or TNF-α. Inhibition of these factors is known to inhibit the inflammatory response (Baeuerle et. al . , Annu. Rev. Immunol., 12: 141-179, 1994). Therefore, the search for a substance that inhibits the production of TNF-α, NO, prostaglandin or IL-1β may prove to be an effective substance for inflammatory diseases such as edema or dermatitis.

Non-steroidal anti-inflammatory drugs (NSAIDS), a widely used drug for the treatment of most inflammatory diseases, are produced from prostaglandin from arachidonic acid called cyclooxygenase (COX). By inhibiting the enzyme activity involved in the biosynthesis of the drug, it exhibits anti-inflammatory action, which causes serious side effects such as gastrointestinal disorders, liver disorders, and renal disorders in addition to the main therapeutic effects, and thus is restricted in long-term use (Rajakariar R et. al. 2006). Therefore, the development of a new anti-inflammatory analgesic that is excellent in anti-inflammatory efficacy without any difficulty for long-term use with little side effects is widely required, which is also a reason for the recent active research on material development through verification of efficacy from natural resources.

Asparagi Tuber is a herbaceous perennial herbaceous plant belonging to the genus Liliaceae Asparagus. cochinchinensis (Lour.) Merr. Is a myocardium, which is mainly used for respiratory diseases and skin diseases due to its effects such as consonant, rhythm, green lung, and jinjin. (Cooperation teaching materials compilation committee, 2005), and it can be used as an ingredient in folk sake. It is native to China and is distributed on the coastal area of the southern part of the Korean peninsula. It is estimated that the seeds of native Chinese species are crowded by the ocean currents and form colonies on the coast of Korea, and are indirectly verified by molecular biological experiments. About the weakness of Astronomical dong in oriental medicine, it is known that it is sweet and bitter and its nature is very cold. Dongbogam describes that astronomical dong helps lung function and is used for various diseases such as cough, stroke and diuresis (Chinese Medicine Dictionary, 1986; Ungokcho Elementary School, 2004; People's Republic of China Pharmacopoeia, 2005). However, the inhibitory effect on fractions fractionated from astronomical extracts is not known.

Thus, the present inventors while studying to develop a drug for inflammatory diseases derived from natural products without toxicity and side effects, the fraction prepared by fractionating the astronomical extract with an organic solvent is a cell that inhibits the production or secretion of inflammatory mediator cytokine The present invention was completed by revealing that it can be usefully used in a composition for preventing or treating inflammatory diseases because it is a non-toxic safe substance.

An object of the present invention to provide a composition for the prevention and treatment of inflammatory diseases containing a fraction prepared from the astronomical extract as an active ingredient.

In order to achieve the above object, the present invention provides a composition for the prevention and treatment of inflammatory diseases containing a fraction prepared by fractionating the extract of Asparagus cochinchinensis with an organic solvent as an active ingredient.

In addition, it provides a health functional food for the prevention and improvement of inflammatory diseases containing the fraction prepared by fractionating the cheonmundong extract with an organic solvent as an active ingredient.

In addition, the present invention provides a functional feed additive for the prevention and improvement of inflammatory diseases containing the fraction prepared by fractionating the astronomical extract with an organic solvent as an active ingredient.

In addition, the present invention provides a cosmetic composition for the prevention and improvement of inflammatory diseases containing a fraction prepared by fractionating the astronomical extract with an organic solvent as an active ingredient.

Asparagus of the present invention cochinchinensis ) fractions are effective in inhibiting the production or secretion of cytokines, which are inflammatory mediators, and are non-cytotoxic, safe substances with excellent anti-inflammatory effects, and thus are useful as active ingredients in compositions for the prevention or treatment of inflammatory diseases. Can be used.

1 is a graph showing the cytotoxicity of the astronomical extract or fractions thereof of the present invention:
70% EtOH Ex: 70% Ethanol Extract;
Hx Fr: n-hexane fraction;
EA Fr: ethyl acetate fraction;
H ₂ 0 Fr: water fraction;
100% H ₂ 0 Fr: 100% water fraction;
60% MeOH Fr: 60% methanol fractions;
90% MeOH Fr: 90% methanol fractions; And
100% MeOH Fr: 100% Methanol Fraction.
Figure 2 is a graph showing the inhibitory effect of interleukin-1β (interleukin-1β, IL-1β) production of the astronomical extract or fractions thereof of the present invention.
Figure 3 is a graph showing the tumor necrosis factor-α (tumor necrosis factor-α, TNF-α) production inhibitory effect of the astronomical extract or fractions thereof of the present invention.

Hereinafter, terms used in the present invention will be described.

As used herein, the term "prevention" means any action that inhibits or delays the progression of an inflammatory disease by administration of a composition of the present invention.

As used herein, the terms "treatment" and "improvement" refer to any action in which an inflammatory disease is improved or beneficially altered by administration of a composition of the present invention.

As used herein, the term "administration" means providing a subject with any of the compositions of the present invention in any suitable manner.

As used herein, the term "individual" refers to any animal, such as a human, monkey, dog, goat, pig or rat, having a disease in which an inflammatory disease can be improved by administering the composition of the present invention.

As used herein, the term “pharmaceutically effective amount” means an amount sufficient to treat a disease at a reasonable benefit or risk ratio applicable to medical treatment, which means the type of disease, the severity, the activity of the drug, the drug Sensitivity to, time of administration, route of administration and rate of administration, duration of treatment, factors including drug used concurrently, and other factors well known in the medical arts.

Hereinafter, the present invention will be described in detail.

The present invention is the Asparagus Cochinchinensis ) provides a composition for the prevention and treatment of inflammatory diseases containing the fraction prepared by fractionating the extract with an organic solvent as an active ingredient.

The fraction is n-hexane fraction, methylene chloride fraction, ethyl acetate fraction, n-butanol fraction or water fraction obtained by systematically fractionating the astronomical extract of n-hexane, methylene chloride, ethyl acetate, n-butanol and water. It is more preferred that the n-hexane fraction or ethyl acetate fraction, but is not limited thereto.

The fraction is 100% water fraction, 30% methanol fraction, 60% methanol fraction, 90% obtained by systematically fractionating the astronomical extract of 100% water, 30% methanol, 60% methanol, 90% methanol and 100% methanol. It is preferably a methanol fraction or 100% methanol fraction and more preferably, but not limited to, 60% methanol fraction, 90% methanol fraction or 100% methanol fraction.

The fraction preferably inhibits the production of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α), but is not limited thereto.

The inflammatory diseases include dermatitis, allergy, atopic dermatitis, asthma, conjunctivitis, periodontitis, rhinitis, otitis media, sore throat, tonsillitis, pneumonia, gastric ulcer, gastritis, Crohn's disease, colitis, hemorrhoids, gout, ankylosing spondylitis, rheumatic fever, lupus, fibromyalgia (fibromyalgia) ), Psoriatic arthritis, osteoarthritis, rheumatoid arthritis, periarthritis, tendonitis, hay salt, periarthritis, myositis, hepatitis, cystitis, nephritis, sjogren's syndrome, multiple sclerosis, and acute and chronic inflammatory diseases It is preferably one of them, but is not limited thereto.

The astronomical extract is preferably prepared by a manufacturing method comprising the following steps, but not always limited thereto:

1) extracting by adding an extraction solvent to Asparagus cochinchinensis ;

2) cooling the extract of step 1) and filtering; And

3) drying the filtered extract of step 2) under reduced pressure.

In the above method, the astronomical dong of step 1) can be used without limitation, such as cultivated or commercially available. The astronomical dong can use all of the outpost, leaves, stems, or roots, it is most preferred to use dried roots, but is not limited thereto.

The extraction solvent is preferably water, alcohol or a mixture thereof. As the alcohol, it is preferable to use C ₁ to C ₄ lower alcohols, ethanol or methanol is preferably used as the lower alcohol, more preferably 70% ethanol, but is not limited thereto. As the extraction method, hot water extraction, immersion extraction, reflux cooling extraction, and ultrasonic extraction may be used, and the extraction may be preferably performed one to five times by the hot water extraction method, and more preferably three times of extraction, but is not limited thereto. . The extraction solvent is preferably extracted by adding 5 to 15 times the weight of the dried astronomical copper root, and more preferably by adding 10 times. The extraction temperature is preferably 40 to 80 ° C and more preferably 50 to 60 ° C, but is not limited thereto. In addition, the extraction time is preferably 2 to 6 hours, more preferably 2 hours 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. In addition, drying may be used under reduced pressure drying, vacuum drying, boiling drying, spray drying or lyophilization, preferably lyophilization, but is not limited thereto.

In a specific embodiment of the present invention, astronomical roots were washed with water and then dried for 7 days at shade and room temperature. The dried Cheonmundong root was crushed and placed in an extraction container, and extracted with an extraction solvent at 50 ~ 60 ℃ for 2 hours. After obtaining the cheonmundong extract, the solids were removed using a filter paper and the suspension was centrifuged to filter the supernatant under reduced pressure. The extract was concentrated under reduced pressure and lyophilized to prepare an astronomical extract.

The fraction of the cheonmundong extract is preferably prepared by a manufacturing method including the step of preparing a fraction by adding a solvent to the cheonmundong extract, but is not limited thereto.

The fraction may be obtained by repeating the fractionation process from 1 to 5 times, preferably 3 times, from the astronomical extract, preferably concentrated under reduced pressure after the fraction, but is not limited thereto.

In a specific embodiment of the present invention, the astronomical copper extract is suspended in distilled water, and then systematically divided into a solvent having different polarities in order of n-hexane, methylene chloride, ethyl acetate, n-butanol and water, and each solvent fraction is decompressed. Concentration prepared n-hexane fraction, methylene chloride fraction, ethyl acetate fraction, n-butanol fraction and water fraction from the astronomical extract.

In a specific embodiment of the present invention, the cheonmundong extract is suspended in methanol, and then systematically fractionated in the order of water, 30% methanol, 60% methanol, 90% methanol and 100% methanol, which are solvents having different polarities. The mixture was concentrated under reduced pressure to prepare 100% water fraction, 30% methanol fraction, 60% methanol fraction, 90% methanol fraction and 100% methanol fraction from the astronomical extract.

In order to confirm the cytotoxicity of the astronomical dong extract or its fraction, the present inventors treated the macrophage line with the 70% ethanol extract or its fraction and determined the cell viability by measuring the absorbance. As a result, there was no significant difference in cell viability compared to the control or cheonmundong extract or fraction. Therefore, it can be seen that the astronomical extract of the present invention or a fraction thereof is a safe substance without cytotoxicity (see FIG. 1).

In order to confirm the inhibitory effect of the astronomical dong extract or fractions thereof, the present inventors have investigated interleukin 1β, an inflammatory cytokine, and tumor necrosis factor- in macrophage lines. α, TNF-α) production inhibition effect was examined. As a result, the astronomical dong extract or fractions thereof inhibited the production and secretion of IL-1β and TNF-α, and in particular, the inhibitory activity of the production of inflammatory cytokines in the fractions was significantly superior to the extracts (FIGS. 2 to 3). Reference). Therefore, the astronomical dong extract of the present invention or a fraction thereof has anti-inflammatory activity, and in particular, it can be seen that the fraction of the astronomical dong extract has an excellent anti-inflammatory effect.

Thus, the fraction prepared from the astronomical extract of the present invention can be usefully used as an active ingredient of the composition for the prevention and treatment of inflammatory diseases.

The pharmaceutical compositions of the present invention may be used in the form of oral dosage forms, such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, external preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods.

Solid form preparations for oral administration include powders, granules, tablets, capsules, soft capsules, and the like. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include powders, granules, tablets, capsules, sterile aqueous solutions, solutions, non-aqueous solutions, suspensions, emulsions, syrups, suppositories, aerosols, etc. It may be used in the form of a formulation, and preferably, an external skin pharmaceutical composition of cream, gel, patch, spray, ointment, warning agent, lotion agent, linen agent, pasta agent or cataplasma agent may be prepared and used. It is not limited to this. Non-aqueous solvents and suspending agents may be used as vegetable oils such as propylene glycol pharmaceutical polyethylene glycol pharmaceutical olive oil, injectable esters such as ethyl oleate, etc. As a base of suppositories, witepsol, macrogol, Tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The pharmaceutical composition according to the present invention may further include conventionally used carriers, excipients, disintegrants, sweeteners, lubricants, flavoring agents and diluents. The carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline Cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. The disintegrants include sodium starch glycolate, crospovidone, croscarmellose sodium, alginic acid, calcium carboxymethyl cellulose, sodium carboxymethyl cellulose, chitosan, guar gum, low-substituted hydroxypropyl cellulose, magnesium aluminum silicate, and polyacryline Potassium and the like.

In addition, the pharmaceutical composition according to the present invention may further include a pharmaceutically acceptable additive, wherein the pharmaceutically acceptable additive may include starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, Calcium hydrogen phosphate, lactose, mannitol, malt, gum arabic, pregelatinized starch, corn starch, powdered cellulose, hydroxypropyl cellulose, opadry, sodium starch glycolate, carnauba lead, synthetic aluminum silicate, stearic acid, magnesium stearate, Aluminum stearate, calcium stearate, sucrose, dextrose, sorbitol, talc and the like can be used. The pharmaceutically acceptable additive according to the present invention is preferably included 0.1 to 90 parts by weight based on the pharmaceutical composition.

The pharmaceutical composition of the present invention may further contain one or more active ingredients exhibiting the same or similar functions in addition to the above components. The composition of the present invention may comprise from 0.0001 to 10% by weight, preferably 0.001 to 1% by weight of the fraction of the present invention based on the total weight of the composition.

In addition, the pharmaceutical composition of the present invention may be administered orally or parenterally, and when parenteral administration is selected for external or intraperitoneal injection, rectal injection, subcutaneous injection, intravenous injection, intramuscular injection or intrathoracic injection injection method. It is desirable to.

 The dosage of the composition of the present invention varies depending on the weight, age, sex, health status, diet, time of administration, administration method, excretion rate and severity of the disease of the patient, the daily dosage is the amount of the extract of astronomical dong 0.0001 to 100 mg / kg, preferably 0.001 to 10 mg / kg, and may be administered 1 to 6 times per day.

The composition of the present invention may be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy and biological response modifiers for the prevention or treatment of inflammatory diseases.

In addition, the present invention provides a method for preventing and treating inflammatory diseases, comprising administering to a subject a fraction prepared by fractionating a pharmaceutically effective amount of astronomical extract in an organic solvent.

The subject is a vertebrate and preferably a mammal, more preferably an experimental animal such as a rat, rabbit, guinea pig, hamster, dog, cat, and most preferably an ape-like animal such as a chimpanzee or gorilla.

The method of administration may be administered orally or parenterally, intraperitoneal, rectal, subcutaneous, intravenous, intramuscular, intrauterine dural, intracerebroventricular or intrathoracic It can be administered by injection.

The pharmaceutically effective amount is 0.0001 to 100 mg / kg, preferably 0.001 to 10 mg / kg, but is not limited thereto. The dose may vary depending on the weight, age, sex, health condition, diet, administration period, method of administration, rate of elimination, severity of disease, and the like of a particular patient.

The inflammatory diseases include dermatitis, allergy, atopic dermatitis, asthma, conjunctivitis, periodontitis, rhinitis, otitis media, sore throat, tonsillitis, pneumonia, gastric ulcer, gastritis, Crohn's disease, colitis, hemorrhoids, gout, ankylosing spondylitis, rheumatic fever, lupus, fibromyalgia (fibromyalgia) ), Psoriatic arthritis, osteoarthritis, rheumatoid arthritis, periarthritis, tendonitis, hay salt, periarthritis, myositis, hepatitis, cystitis, nephritis, sjogren's syndrome, multiple sclerosis, and acute and chronic inflammatory diseases It is preferably one of them, but is not limited thereto.

Fractions prepared from the astronomical extract of the present invention inhibit the production or secretion of the inflammatory cytokines, interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) Since it is a safe substance without cytotoxicity, it can be usefully used for the prevention or treatment of inflammatory diseases.

In addition, it provides a health functional food for the prevention and improvement of inflammatory diseases containing the fraction prepared by fractionating the cheonmundong extract with an organic solvent as an active ingredient.

The inflammatory diseases include dermatitis, allergy, atopic dermatitis, asthma, conjunctivitis, periodontitis, rhinitis, otitis media, sore throat, tonsillitis, pneumonia, gastric ulcer, gastritis, Crohn's disease, colitis, hemorrhoids, gout, ankylosing spondylitis, rheumatic fever, lupus, fibromyalgia (fibromyalgia) ), Psoriatic arthritis, osteoarthritis, rheumatoid arthritis, periarthritis, tendonitis, hay salt, periarthritis, myositis, hepatitis, cystitis, nephritis, sjogren's syndrome, multiple sclerosis, and acute and chronic inflammatory diseases It is preferably one of them, but is not limited thereto.

Fractions prepared from the astronomical extract of the present invention can be added as is or used with other foods or food ingredients, and can be suitably used according to conventional methods.

There is no particular limitation on the kind of the food. Examples of the food to which the cheonmundong extract or a fraction thereof may be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, Tea, drink, alcoholic beverages and vitamin complexes, etc., and includes all of the health food in the usual 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 proportion of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 ml of the composition of the present invention.

In addition to the above, fractions of the astronomical extract of the present invention are various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH regulators, stabilizers, preservatives, glycerin , Alcohols, carbonating agents 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.

Fractions prepared from the astronomical extract of the present invention inhibit the production or secretion of the inflammatory cytokines, interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) And because it is a safe substance without cytotoxicity, it can be usefully used in the health functional food for the prevention or improvement of inflammatory diseases.

In addition, the present invention provides a functional feed additive for the prevention and improvement of inflammatory diseases containing the fraction prepared by fractionating the astronomical extract with an organic solvent as an active ingredient.

The inflammatory diseases include dermatitis, allergy, atopic dermatitis, asthma, conjunctivitis, periodontitis, rhinitis, otitis media, sore throat, tonsillitis, pneumonia, gastric ulcer, gastritis, Crohn's disease, colitis, hemorrhoids, gout, ankylosing spondylitis, rheumatic fever, lupus, fibromyalgia (fibromyalgia) ), Psoriatic arthritis, osteoarthritis, rheumatoid arthritis, periarthritis, tendonitis, hay salt, periarthritis, myositis, hepatitis, cystitis, nephritis, sjogren's syndrome, multiple sclerosis, and acute and chronic inflammatory diseases It is preferably one of them, but is not limited thereto.

 The feed additives can be prevented by steadily ingesting poultry, livestock, and the like, and can treat inflammation that has already occurred.

In the feed additive of the present invention, 0.1 to 20% by weight of the fraction of the cheonmundong extract, 0.001 to 0.01% by weight of lipase, 1 to 20% by weight of tricalcium phosphate, and 0.01 to 0.01% of vitamin E 0.1 parts by weight, enzyme powder 1 to 10% by weight, lactic acid bacteria 0.1 to 10% by weight, Bacillus (Bacillus) culture medium is preferably composed of 0.01 to 10% by weight and glucose 20 to 90% by weight However, the present invention is not limited thereto, and if the fraction of the astronomical extract is added in an effective amount, it can be used as a feed additive of the present invention.

The effective amount means an amount capable of preventing inflammatory diseases or treating inflammatory diseases that have already occurred by allowing poultry, livestock, etc. to be continuously ingested. Moreover, it is preferable that the quantity which does not produce the bad influence beyond the benefit by addition is preferable.

In addition, the feed additive may additionally contain a carrier that is acceptable to poultry and livestock. In the present invention, the feed additive may be added as it is or a known carrier, stabilizer and the like, and various nutrients such as vitamins, amino acids and minerals, antioxidants, antibiotics, antibacterial agents and other additives may be added as necessary. The shape may be in a suitable state such as powder, granules, pellets, suspension, or the like. When supplying the feed additive of the present invention can be supplied alone or mixed in the feed for poultry and livestock.

Fractions prepared from the astronomical extract of the present invention inhibit the production or secretion of the inflammatory cytokines, interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) Because it is a safe substance without cytotoxicity, it can be usefully used in functional feed additives for the prevention or improvement of inflammatory diseases.

In addition, the present invention provides a cosmetic composition for the prevention and improvement of inflammatory diseases containing a fraction prepared by fractionating the astronomical extract with an organic solvent as an active ingredient.

The inflammatory diseases include dermatitis, allergy, atopic dermatitis, asthma, conjunctivitis, periodontitis, rhinitis, otitis media, sore throat, tonsillitis, pneumonia, gastric ulcer, gastritis, Crohn's disease, colitis, hemorrhoids, gout, ankylosing spondylitis, rheumatic fever, lupus, fibromyalgia (fibromyalgia) ), Psoriatic arthritis, osteoarthritis, rheumatoid arthritis, periarthritis, tendonitis, hay salt, periarthritis, myositis, hepatitis, cystitis, nephritis, sjogren's syndrome, multiple sclerosis, and acute and chronic inflammatory diseases It is preferably any one, more preferably edema or dermatitis, but is not limited thereto.

Cosmetics prepared by containing the fraction of the cheonmundong extract of the present invention as an active ingredient can be prepared in the form of a general emulsion formulation and solubilized formulation. Cosmetics of emulsified form include nutrition lotion, cream, essence, etc., and cosmetics of solubilized form have softening longevity. Suitable cosmetic formulations include, for example, solutions, gels, solid or kneaded anhydrous products, emulsions obtained by dispersing the oil phase in water, suspensions, microemulsions, microcapsules, microgranules or ionic (liposomes) In the form of a dispersing agent, in the form of a cream, a skin, a lotion, a powder, an ointment, a spray or a conical stick. It may also be prepared in the form of a foam or in the form of an aerosol composition further containing a compressed propellant.

In addition to the fractions of the extract of the present invention, the cosmetics, in addition to fatty substances, organic solvents, solubilizers, thickening and gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, Commonly used in water, ionic or nonionic emulsifiers, fillers, metal ion sequestrants and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or cosmetics And any other ingredients that may be used, such as auxiliaries commonly used in the cosmetic field.

Fractions prepared from the astronomical extract of the present invention inhibit the production or secretion of the inflammatory cytokines, interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) And because it is a safe material without cytotoxicity, it can be usefully used in the cosmetic composition for the prevention or improvement of inflammatory diseases.

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

However, the following Examples and Preparation Examples are merely illustrative of the present invention, and the content of the present invention is not limited to the following Examples and Preparation Examples.

Astronomical building  Preparation of extract

Asparagus collected from Gyehwa-do, Jeollabuk-do cochinchinensis ) roots were dried and ground. 100 g of astronomical dong prepared by the above method was immersed in 1 L of 70% ethanol aqueous solution (70% EtOH), 100% ethanol (100% EtOH), 100% methanol (100% MeOH) or water (H ₂ O) and stirred. Then, the mixture was heated to reflux for 2 hours at an extraction temperature maintained at 50 to 60 ° C. and filtered through a filter paper. The extract was concentrated under reduced pressure using a concentrator at 50 to 55 ° C., and then freeze-dried to prepare 70% ethanol, 100% ethanol, 100% methanol, and water extracts of astronomical dong.

As a result, astronomical copper solvent extracts of 70% ethanol (15 g), 100% ethanol (15 g), 100% methanol (14 g) and water (17 g) were obtained, respectively.

Astronomical building  From the extract Fraction  Produce

<2-1> organic solvent Fraction  Produce

In order to prepare a fraction from the astronomical dong extract, 1760 kg of 70% ethanol extract was obtained from 11.4 kg of dried astronomical dong root according to the method of <Example 1>. After 1,760 g of the extract was suspended in 5 L of water (H ₂ O), the suspension and the same amount of n-hexane (n-hexane, Hx) were added thereto, and the mixture was left to separate into an aqueous layer and an Hx layer. After the chemical equilibrium was separated, the Hx layer was separated and a new amount of Hx was poured again and allowed to separate into the water layer and the Hx layer. The Hx layer obtained by repeating the layer separation process three times was concentrated under reduced pressure to obtain an Hx fraction (0.9 g). In the same manner as above, the Hx fractions were separated and the remaining water layer was separated from methylene chloride (methylene chloride, dichloromethane, MC), ethyl acetate (EA), n-butanol (n-butanol, BuOH), Systematic fraction extraction was performed in the order of water (H ₂ O) to prepare astronomical copper methylene chloride, ethyl acetate, n-butanol and water fractions.

As a result, MC (3 g), EA (3 g), BuOH (29 g) and H ₂ O (680 g) astronomical copper fractions were obtained, respectively.

<2-2> methanol Fraction  Produce

In order to prepare a fraction from the astronomical dong extract, 80 g of 70% ethanol extract was obtained from 500 g of dried astronomical dong root according to the method of <Example 1>. 80 g of the extract was suspended in 0.5 L of methanol (MeOH), followed by 100% water (100% H ₂ O) and different proportions (30%, 60%, 90% and 100%) using Amberlite XAD-4. Fraction extraction with methanol, followed by 100% water fraction (100% H ₂ O, 34 g), 30% MeOH fraction (30% MeOH, 1.3 g), 60% MeOH fraction (60% MeOH, 0.2 g). ), 90% MeOH fractions (90% MeOH, 0.2 g) and 100% MeOH fractions (100% MeOH, 0.03 g) were obtained, respectively.

Cytotoxicity experiment

In order to confirm the safety of the astronomical extract or the fraction thereof prepared in <Example 1> and <Example 2>, cytotoxicity was confirmed by MTT analysis. Mouse macrophage RAW264.7 cell line was purchased from American Type Culture Collection (ATCC, Rockville, MD, USA) to obtain 10% fetal bovine serum (FBS), 100 units / ml penicillin And 100 μg / ml streptomycin-added DMEM medium in a 37 ° C., 5% CO ₂ environment. After treatment with astronomical dong extract or fractions in a RAW 264.7 macrophage by concentration and incubated for 24 hours, 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide {3- ( 4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide, MTT} 500 μg / ml was added and reacted at 37 ° C. for 1 hour. After the reaction, all of the culture medium was removed and dimethyl sulfoxide (DMSO) solution was added to dissolve the formazan produced. The absorbance was measured at 570 nm. Relative cell viability (% of control) was calculated by comparing the measured value with the control.

As a result, as shown in Figure 1, astronomical dong 70% ethanol extract, n-hexane fraction (Hx), ethyl acetate fraction (EA), water fraction (H ₂ O), 100% water fraction (100% H ₂ O) , 60% methanol fraction (60% MeOH), 90% methanol fraction (90% MeOH) or 100% methanol fraction (100% MeOH) showed no significant difference in cell viability compared to the control at all concentrations. Therefore, the astronomical extract or fractions thereof of the present invention was confirmed that the material is not cytotoxic (Fig. 1).

Inflammatory cytokines ( cytokine ) Suppression effect

In order to investigate the inhibitory effect of the astronomical dong extract of the present invention or its fraction on the inflammation-inducing factor, interleukin-1β, which is an inflammatory cytokine, and tumor necrosis factor-α , TNF-α) was confirmed. Specifically, 1 μg / ml lipopolysaccharide (LPS) and 100 μg / ml astronomical dong extract or fractions were treated in RAW264.7 macrophage line and cultured for 20 hours, and then IL-1β and TNF secreted into the medium. The amount of -α was analyzed by ELISA kit (R & D Systems Inc., Minneapolis, MN, USA). The 100% production amount was defined as the LPS-treated group to calculate the relative% of control of the astronomical sample treatment group. All measurements were expressed as mean and standard deviation (SD), and the differences between the groups were determined by statistical analysis using Student's t-test.

As a result, as shown in FIG. 2, in the case of IL-1β, the IC ₅₀ of the astronomical extract was 1209.63 µg / ml. In contrast, the IC ₅₀ of the fraction was 321.95 μg / ml for the n-hexane fraction (Hx), 326.53 μg / ml for the ethyl acetate fraction (EA), and 375.20 μg / ml for the 60% methanol fraction (60% MeOH). The% methanol fraction (90% MeOH) was 81.57 μg / ml and the 100% methanol fraction (100% MeOH) was 23.08 μg / ml, about 3.8, 3.7, 3.2, 15, and 52 times higher than the extract, respectively. It showed a significant IL-1β inhibitory effect. On the other hand, the IL-1β inhibitory effect of the water fraction (H ₂ O) or 100% water fraction (100% H ₂ O) was lower than the extract (Fig. 2). In addition, as shown in FIG. 3, in the case of TNF-α, the IC ₅₀ of the astronomical extract was 1027.33 μg / ml. In contrast, the IC ₅₀ of the fractions was 358.58 μg / ml for the n-hexane fraction (Hx), 297.08 μg / ml for the ethyl acetate fraction (EA), and 898.74 μg / ml for the 60% methanol fraction (60% MeOH). The methanol fraction (90% MeOH) is 672.12 μg / ml and the 100% methanol fraction (100% MeOH) is 103.40 μg / ml, about 2.9, 3.5, 1.1, 1.5 and 10 times better than the extract, respectively. TNF-α inhibitory effect was shown, whereas the TNF-α inhibitory effect of water fraction (H ₂ O) or 100% water fraction (100% H ₂ O) was lower than the extract (see FIG. 3). Therefore, the astronomical extract of the present invention or fractions thereof inhibits the production and secretion of inflammatory cytokines IL-1β and TNF-α, and in particular, the inflammatory cytokine inhibitory activity of the fractions is remarkably superior to have an inflammatory response inhibitory effect. Confirmed.

< Manufacturing example  1> Preparation of Pharmaceutical Formulations

<1-1> Sanje  Produce

2 g of ethyl acetate fraction of Example <2-1> 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 ethyl acetate fraction of Example <2-1> 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 butanol fraction of Example <2-1> 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 butanol fraction of Example <2-1> of the present invention

Lactose 1.5 g

1 g of glycerin

Xylitol 0.5 g

After mixing the above components, it was prepared to be 4 g per ring according to a conventional method.

<1-5> Preparation of granules

150 mg of butanol fraction of Example <2-1> 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> Manufacturing of food

Food containing a fraction of the cheonmundong extract of the present invention was prepared as follows.

<2-1> Production of flour food

0.5-5.0 parts by weight of the n-hexane fraction of Example <2-1> of the present invention was added to the flour, and bread, cake, cookies, crackers and noodles were prepared using this mixture.

<2-2> soup  And juicy ( gravies Manufacturing

0.1-5.0 parts by weight of the n-hexane fraction of Example <2-1> of the present invention was added to soups and broth to prepare meat products for health promotion, soups of noodles, and broth.

<2-3> Ground Beef  Produce

10 parts by weight of the n-hexane fraction of Example <2-1> of the present invention was added to the ground beef to prepare a ground beef for health promotion.

<2-4> Dairy products ( dairy products Manufacturing

5-10 parts by weight of the n-hexane fraction of Example <2-1> of the present invention was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

<2-5> 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 ethyl acetate fraction of Example <2-1> of the present invention was concentrated under reduced pressure in a vacuum concentrator, and the dried product obtained by spraying and drying with a hot air dryer was pulverized with a particle size of 60 mesh to obtain a dry powder.

The grains, seeds and the ethyl acetate fractions of Example <2-1> prepared above were prepared by combining the following ratios:

(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)

Ethyl acetate fraction of Example <2-1> of the present invention (3 parts by weight),

(0.5 parts by weight), and

Rhubarb (0.5 parts by weight).

< Manufacturing example  3> Manufacturing of beverage

<3-1> Health drink  Produce

Substances such as liquid fructose (0.5%), oligosaccharides (2%), sugars (2%), salts (0.5%) and water (75%) and 5 g of ethyl acetate fraction of Example <2-1> of the present invention After homogeneous mixing and sterilization, it was prepared by packaging in a small packaging container such as glass bottles, plastic bottles.

<3-2> Preparation of vegetable juice

Vegetable juice was prepared by adding 5 g of the n-hexane fraction of Example <2-1> of the present invention to 1,000 ml of tomato or carrot juice.

<3-3> Production of fruit juice

1 g of the n-hexane fraction of Example <2-1> of the present invention was added to 1,000 ml of apple or grape juice to prepare a fruit juice.

< Manufacturing example  4> Preparation of feed additive

The present inventors prepared the feed additive with the following composition using the butanol fraction of Example <2-1> as an active ingredient.

<The composition of the feed additive>

Butanol fraction of Example <2-1> of the present invention: 0.1 to 20% by weight,

Lipase: 0.001 to 0.01% by weight,

Tertiary calcium phosphate: 1-20% by weight,

Vitamin E: 0.01-0.1% by weight,

Enzyme powder: 1 to 10% by weight,

Lactic acid bacteria: 0.1 to 10% by weight,

Bacillus culture medium: 0.01 to 10% by weight, and

Glucose: 20 to 90% by weight.

< Manufacturing example  5> Manufacture of cosmetics

It is possible to prepare cosmetics for the prevention and improvement of inflammatory diseases containing a fraction of the astronomical extract of the present invention as an active ingredient. The present inventors prepared the cosmetic of solubilized formulations such as cosmetics and soft cosmetics of the emulsion formulations such as nutrient cosmetics, creams, essences and the like as the cosmetics.

<5-1> Cosmetic Preparation of Emulsion Formulation

To the cosmetic composition of the emulsifier type was prepared in the composition shown in Table 1. The manufacturing method is as follows.

1) The mixture which mixed the raw materials of 1-9 was heated at 65-70 degreeC.

2) 10 was added to the mixture of step 1).

3) The mixture of the raw materials of 11-13 was heated to 65-70 degreeC, and was fully dissolved.

4) While passing through the step 3), the mixture of 2) was slowly added to emulsify for 2-3 minutes at 8,000 rpm.

5) The raw material of 14 was dissolved in a small amount of water, and then added to the mixture of step 4) and emulsified for 2 minutes.

6) 15 to 17 of the raw materials were weighed, respectively, and added to the mixture of step 5) and emulsified for 30 seconds.

7) After the emulsification of the mixture of step 6) through a degassing process was cooled to 25 ~ 35 ℃ to prepare an emulsion cosmetic.

Composition of Emulsifying Formulations 1, 2, 3 Furtherance Emulsifier 1 Emulsifier 2 Emulsifier 3 One Stearic acid 0.3 0.3 0.3 2 Stealy alcohol 0.2 0.2 0.2 3 Glyceryl Monostearate 1.2 1.2 1.2 4 Wax 0.4 0.4 0.4 5 Polyoxyethylene sorbitan
Monolauric acid ester 2.2 2.2 2.2 6 Methyl paraoxybenzoate 0.1 0.1 0.1 7 P-hydroxybenzoic acid profile 0.05 0.05 0.05 8 Cetylethylhexanoate 5 5 5 9 Triglyceride 2 2 2 10 Cyclomethicone 3 3 3 11 Distilled water ~ 100 ~ 100 ~ 100 12 Concentrated glycerin 5 5 5 13 Triethanolamine 0.15 0.15 0.15 14 Polyacrylic acid polymer 0.12 0.12 0.12 15 Pigment 0.0001 0.0001 0.0001 16 incense 0.10 0.10 0.10 17 60% Methanol Fraction of Example <2-2> 0.0001 One 10

<5-2> Solubilization  Cosmetic preparation of the formulation

A cosmetic of a solubilized formulation was prepared with the composition shown in Table 2 below. The manufacturing method is as follows.

1) Raw materials 2 to 6 were placed in raw material 1 (purified water) and dissolved using a still mixer.

2) The raw materials of 8 to 11 were put into the raw material of 7 (alcohol) and completely dissolved.

3) The mixture of step 2) was solubilized with slow addition to the mixture of step 1).

Composition of Solubilized Formulations 1, 2, 3 Furtherance Solubilized Formulation 1 Solubilized Formulation 2 Solubilized Formulation 3 One Purified water ~ 100 ~ 100 ~ 100 2 Concentrated glycerin 3 3 3 3 1,3-butylene glycol 2 2 2 4 EDTA-2Na 0.01 0.01 0.01 5 Pigment 0.0001 0.0002 0.0002 6 60% Methanol Fraction of Example <2-2> 0.1 5 5 7 Alcohol (95%) 8 8 8 8 Methyl paraoxybenzoate 0.1 0.1 0.1 9 Polyoxyethylene
Hydrogenated Ester 0.3 0.3 0.3 10 incense 0.15 0.15 0.15 11 Cyclomethicone - - 0.2

As described above, the fraction prepared by the system fractionation of the cheonmundong extract of the present invention has an excellent effect on inhibiting inflammation, so that the drugs related to the prevention or treatment of inflammatory diseases, or functional health foods for improving inflammation, functional feed additives or cosmetic compositions It can be usefully used for development and production.

Claims (13)

A fraction prepared by fractionation of Asparagus cochinchinensis extract with an organic solvent is contained as an active ingredient.
N-hexane fraction, ethyl acetate fraction and n-butanol fraction obtained by systematically fractionating the astronomical extract from n-hexane, methylene chloride, ethyl acetate, n-butanol and water; And
At least one fraction selected from the group consisting of 90% methanol fraction and 100% methanol fraction obtained by sequential fractionation of 100% water, 30% methanol, 60% methanol, 90% methanol and 100% methanol. Composition for the prevention and treatment of inflammatory diseases.
The method of claim 1, wherein the astronomical extract is water, C ₁ A composition for the prevention and treatment of inflammatory diseases, which is extracted with a lower alcohol of C ₄ or a mixed solvent thereof.
According to claim 2, wherein the lower alcohol is a composition for the prevention and treatment of inflammatory diseases, characterized in that ethanol or methanol.
delete [Claim 2] The inflammatory disease of claim 1, wherein the fraction is an n-hexane fraction or an ethyl acetate fraction obtained by sequential fractionation of the astronomical extract of n-hexane, methylene chloride, ethyl acetate, n-butanol and water. Prophylactic and therapeutic compositions.
delete delete According to claim 1, wherein the fraction is a composition for the prevention and treatment of inflammatory diseases, characterized in that to inhibit the production of interleukin-1β (interleukin-1β, IL-1β).
According to claim 1, wherein the fraction is a composition for the prevention and treatment of inflammatory diseases, characterized in that to suppress the production of tumor necrosis factor-α (TNF-α).
The method of claim 1, wherein the inflammatory disease is dermatitis, allergy, atopic, asthma, conjunctivitis, periodontitis, rhinitis, otitis media, sore throat, tonsillitis, pneumonia, gastric ulcer, gastritis, Crohn's disease, colitis, hemorrhoids, gout, ankylosing spondylitis, rheumatic fever , Lupus, fibromyalgia, psoriatic arthritis, osteoarthritis, rheumatoid arthritis, periarthritis, tendonitis, hay salt, peritonitis, myositis, hepatitis, cystitis, nephritis, sjogren's syndrome, multiple sclerosis, and acute and chronic inflammatory diseases The composition for the prevention and treatment of inflammatory diseases, characterized in that any one selected from the group consisting of.
Health functional food for the prevention and improvement of inflammatory diseases containing the fraction of claim 1 as an active ingredient.
delete delete

Images