KR101487065B1 - A pharmaceutical composition for prevention or treatment of inflammatory disease comprising Myagropsis myagroides extracts or fraction thereof as an effective ingredient - Google Patents

Abstract

The present invention relates to a composition for the treatment or prevention of an inflammatory disease, and provides a pharmaceutical composition for preventing or treating an inflammatory disease containing an extract of Myagropsis myagroides or a fraction of the extract as an active ingredient. Solitary of the present invention more Sargassum extract or its fractions prior known Foucault xanthine (fucoxanthin) inhibiting to nitrogen monoxide (NO), prostaglandin _{E 2 (prostaglandin E 2, PGE} 2) , and production of inflammatory cytokines, does not at all contain In addition, iNOS and COX-2 effectively inhibit expression, and thus can be effectively used for prevention and treatment of inflammatory diseases.

Description

TECHNICAL FIELD The present invention relates to a pharmaceutical composition for preventing or treating an inflammatory disease containing extracts of Mycobacterium tuberculosis or fractions thereof as an active ingredient,

The present invention relates to a composition for treating or preventing an inflammatory disease, and more particularly to a composition for treating or preventing an inflammatory disease, which contains an extract of Solanum tuberculosis or a fraction thereof as an active ingredient.

Inflammation is one of the damage of tissue, external stimulation or defense of biological tissues against various infectious agents. Enzyme activation, inflammation mediator secretion, cell infiltration due to organic interactions of various inflammatory mediators and various immune cells in blood vessels and body fluids And a series of complex pathologies such as fluid exudation, circulatory disturbances, tissue degeneration and hyperplasia. In the process of inflammatory reaction, macrophages are collected at the wound area at the initial stage and attack the invading bacteria. After that, plasma accumulates on the injured area and blood flow is increased, resulting in external symptoms such as fever, erythema, edema and pain . If the inflammatory reaction continues or occurs excessively, it progresses to a major pathological condition (irritable allergic disease, chronic inflammatory disease) of the disease, resulting in serious abnormal disorder.

Nonsteroidal anti-inflammatory drugs (NSAIDS), a widely used agent for the treatment of most inflammatory diseases, are produced from arachidonic acid called prostaglandin, called cyclooxygenase (COX) (Rajakariar R et < RTI ID = 0.0 > (R) < / RTI > et < RTI ID = 0.0 > al., 2006). Therefore, the development of a novel anti-inflammatory analgesic agent that is excellent in anti-inflammatory efficacy is widely demanded because there is no side effect and it is not difficult to use for a long period of time. This is why research on material development through the verification of efficacy from natural resources is being activated recently.

Various biochemical phenomena are involved in the cause of inflammation in living body. Macrophage is a cell with various functions. It produces various cytokines and NO by chemical stimulation and plays an important role in the inflammation reaction. In particular, inducible nitric oxide synthase (iNOS) expressed by macrophage-stimulated cytokines such as lipopolysaccharide (LPS) and interferon gamma and TNF-α produces large amounts of nitrogen oxides (NO) over a long period of time. These oxidative stresses are known to promote NFκB activity, a transcription factor of inflammatory responses inhibited by IκB. Activated NFκB is known to migrate to the nucleus and promote gene expression that induces inflammatory responses such as iNOS, COX-2, and several cytokines such as IL-1β and TNF-α. When these factors are inhibited, (Baeuerle et al., Annu. Rev. Immunol., 12: 141-179, 1994).

Nitrogen oxides (NO) are produced from L-arginine by three major nitrogen oxide synthase (NOS) isomers, neuronal NOS (nNOS), endothelial NOS (eNOS) and inducible NOS (iNOS). While nNOS and eNOS are regulated by Ca2 + / calmodulin, iNOS is regulated at the transcriptional level by inflammatory stimuli such as interleukin, interferon, and LPS. NO produced by nNOS or eNOS plays a normal physiological function such as vasodilation, neurotransmission, cell destruction to pathogen, etc. However, overexpressed NO produced by macrophages in iNOS is involved in various pathophysiological And reacts with superoxide to form peroxynitrite, which acts as a powerful oxidant to damage cells, activates NFκB in macrophages activated by inflammatory stimuli, (Rice et al., 2002), and it is known to be associated with chronic diseases such as cancer, arteriosclerosis (Lawrence et al., Nat Med., 7: 1291-1297, 2001; Riehemann et al., FEBS Lett., 442: et al., Science, 258: 1898-1902, 1992).

Prostaglandins are unsaturated fatty acid derivatives containing 20 carbon atoms with a cyclic structure and are mainly chemical mediators involved in chronic inflammatory diseases and are also involved in autoimmune diseases such as asthma (Ruf et al., Eur. J. Biochem., 204: 1069-1073,1992). Prostaglandins are biosynthesized by COX, which has two isomers of COX-1 and COX-2 (Smith et al., J. Biol. Chem., 271: 33157-33160, 1996). COX-1 is a constant enzyme present in tissues such as the stomach or kidney, and is involved in maintaining normal homeostasis, whereas COX-2 is a transient and rapidly expressed expression in cells by cytokines or cytokines in inflammation or other immune responses do. NSAIDs used in the treatment of chronic inflammatory diseases such as acute or rheumatoid arthritis are known to exhibit various side effects such as the aforementioned gastrointestinal disorders by inhibiting COX-1 enzyme as well as inhibiting COX-2 enzyme Masferrer et al., P. Natl. Acad. Sci. USA., 91: 3228-3232, 1994).

Therefore, when a substance that inhibits the production of tumor necrosis factor (TNF-α) and nitrogen oxide (NO), inhibits prostaglandin production, or inhibits the production of IL-1β is searched, will be.

On the other hand, Myagropsis myagroides is a brown algae belonging to the mongolian mongolica , and lives on the southern coast of Korea and all coastal areas of Japan and is mainly used as feed. Studies have been carried out on the alopecia areata (Lee et al., 2010), antihypertensive activity (Cha et al., 2006), liver injury protection (Wong et al., 2004), anti- (Kim et al., 2007), and the effects of the antioxidant enzymes on the lipid metabolism (Kim et al., 2006) Foucault, isolated from kelp (Lee et al., Eur. J. Nutr., Epub ahead of print, 2012) Anti-inflammatory effects of fucoxanthin have been reported. However, no studies have been conducted on the anti-inflammatory activity of the extracts of Solanum tuberculosis extract containing no fucoxanthin.

It is an object of the present invention to provide a pharmaceutical composition, a health functional food and a cosmetic composition for treating or preventing an inflammatory disease containing an extract of Solanum tuberculosis or a fraction thereof as an active ingredient.

In order to accomplish the above object, one aspect of the present invention is a method for producing Myagropsis myagroides ) as an active ingredient. The present invention also provides a pharmaceutical composition for preventing or treating an inflammatory disease.

In order to achieve the above object, another aspect of the present invention is to provide a method for preventing Myagropsis myagroides ) extract as an active ingredient. The present invention also provides a pharmaceutical composition for preventing or treating an inflammatory disease.

In order to achieve the above-mentioned object, another aspect of the present invention is a method for preventing Myagropsis myagroides extract, or a fraction of the above extract as an active ingredient. The present invention also provides a health functional food for preventing or ameliorating an inflammatory disease.

In order to achieve the above object, another aspect of the present invention is to provide a method for preventing Myagropsis myagroides extract or a fraction of the above extract as an active ingredient. The present invention also provides a cosmetic composition for preventing or ameliorating an inflammatory disease.

Solitary of the present invention more Sargassum extract or its fractions prior known Foucault xanthine (fucoxanthin) inhibiting to nitrogen monoxide (NO), prostaglandin _{E 2 (prostaglandin E 2, PGE} 2) , and production of inflammatory cytokines, does not at all contain In addition, iNOS and COX-2 effectively inhibit expression, and thus can be effectively used for prevention and treatment of inflammatory diseases.

However, the effects of the present invention are not limited to the above-mentioned effects, and other effects not mentioned can be clearly understood by those skilled in the art from the following description.

Fig. 1 is a flowchart showing a process for obtaining an extract (a) and a fraction (b) from an alfalfa solubilis.
2 is a graph showing the results of measurement of the content of the polyphenolic compound contained in the extract of the alfalfa solanaceae.
FIG. 3 is a graph showing the results of measurement of the content of fucoxanthin contained in the standard substance (fucosanthin) (a), alcohol extract (b) and n-hexane fraction (c) of the alcohol extract.
FIG. 4 is a graph showing the results of measurement of cytotoxicity of extracts of Solanum tuberculosis.
FIG. 5 is a graph showing the results of measurement of the inhibitory effect of nitrogen monoxide (NO) production on the extract of Solanum tuberculosis.
FIG. 6 is a graph showing the results of measurement of cytotoxicity of extracts of Solanum tuberosum and various fractions thereof.
FIG. 7 is a graph showing the results of measuring the inhibitory effect of nitrogen monoxide (NO) production on the extracts of alcohol extract from Solanum tuberculosis and various fractions of the above-mentioned alcohol extract.
FIG. 8 shows the inhibitory effect of (a) prostaglandin E ₂ , (b) TNF-α, (c) IL-1β and (d) IL-6 production inhibition of the alcohol extract of Allium tuberculosis and the n-hexane fraction of the alcohol extract Fig.
FIG. 9 is a graph showing the results of measurement of the protein expression of (a) iNOS and COX-2 and (b) the inhibitory effect of iNOS and COX-2 on mRNA expression of the alcohol extract of Solanum tuberculosis and the n-hexane fraction of the above- to be.
FIG. 10 is a graph showing the results of measuring the NF-κB inhibitory effect of the alcohol extract of Solanum tuberculosis and the n-hexane fraction of the alcohol extract.
FIG. 11 is a photograph showing the inhibitory effect of NF-.kappa.B nuclear transfer inhibition of the alcohol extract of Solanum tuberculosis and the ethanol extract of n-hexane by confocal microscope.
12 is a graph showing the results of measuring the inhibitory effect of NF-κB nuclear transfer-related proteins on the extracts of alcohol extracts from Solanum tuberculosis and the n-hexane fraction of the alcohol extracts.
FIG. 13 is a graph showing the results of measuring the inhibitory effect of MAPKs and Akt proteins on the extracts of alcohol extracts from Solanum tuberculosis and the n-hexane fraction of the alcohol extracts.

First, terms used in the specification of the present invention will be described.

The term " inflammation " as used herein refers to the pathological condition of an abscess formed by the infiltration of external infectious agents (bacteria, fungi, viruses, various kinds of allergens).

As used herein, the term " prophylactic " means any action that inhibits or slows the progression of an inflammatory disease upon administration of the composition of the present invention.

As used herein, the terms "improvement" and "treatment" refer to all actions by which administration of the composition of the present invention improves or alleviates the symptoms of an inflammatory disease.

The term " administration " as used herein is meant to provide any desired composition of the invention to a subject in any suitable manner.

The term " individual " as used herein means all animals such as a human, a monkey, a dog, a goat, a pig or a mouse having a disease in which symptoms of an inflammatory disease can be improved by administering the composition of the present invention.

The term " pharmaceutically effective amount " as used herein means an amount sufficient to treat a disease at a reasonable benefit or risk ratio applicable to medical treatment, including the type of disease, severity, activity of the drug, The time of administration, the route and rate of excretion of the drug, the duration of the treatment, factors including drugs used simultaneously and other factors well known in the medical arts.

Hereinafter, the present invention will be described in detail.

One. Pharmaceutical compositions for the prevention or treatment of inflammatory diseases

One aspect of the present invention is a method for treating Myagropsis myagroides ) as an active ingredient. The present invention also provides a pharmaceutical composition for preventing or treating an inflammatory disease.

Further, another aspect of the present invention is more Solitary Sargassum (Myagropsis myagroides ) extract as an active ingredient. The present invention also provides a pharmaceutical composition for preventing or treating an inflammatory disease.

The pharmaceutical composition of the present invention relates to the prevention, improvement or therapeutic use in inflammatory diseases, including the fraction of the extract or extracts of Sargassum Solitary dog Solitary dog Sargassum (Myagropsis myagroides).

The extract can be obtained by various extraction methods known in the art. Preferably, the Solanum tuberculosis extract includes, but is not limited to, 1) Myagropsis myagroides ) and extracting the extract to obtain an extract; 2) filtering the extract obtained in the step 1) to obtain a filtrate; And 3) concentrating the filtrate obtained in the step 2) and drying the filtrate under reduced pressure to obtain an extract.

It is preferable that the above-mentioned step 1) can be used either naturally cultivated or cultured, and it is preferably dried and used regardless of the site. Particularly, in the case of drying and using the above-mentioned solids, the dried material is preferably cut or ground so as to have a large surface area reacted with the extraction solvent.

The extraction solvent of step 1) may be any kind of solvent commonly used in the art, but it is preferable that the extraction solvent of step 1) is water, an organic solvent or a mixture thereof. The extraction solvent of step 1) is preferably a C ₁ to C ₄ lower alcohol, more preferably the extraction solvent of step 1) is methanol or ethanol, and the extraction solvent of step 1) , But not limited to. It is preferable that the extraction solvent of step 1) is added by 2 to 20 times the total weight of the extract, and the extraction is performed by adding 4 to 15 times the total weight of the extract, Most preferably 5 to 7 times the total weight of the outer layer of the outer layer, but is not limited thereto.

The extraction of the above step 1) can be carried out by all commonly used extraction methods such as hot water extraction, hot water extraction, ultrasonic extraction, reflux cooling extraction, Soxhlet extraction, Bligh / Dyer extraction, Stass Otto extraction and Folch extraction . Further, the extraction of the step 1) is carried out at a temperature of 50 to 250 DEG C, preferably 70 to 150 DEG C for 1 to 12 hours, preferably 3 to 10 hours to prevent decay, discoloration and detachment And may be repeated three to five times.

The filtration in the step 2) may be performed using any filtration method commonly used in the art, and may be filtered using a filter paper, but is not limited thereto.

The vacuum concentration in step 3) is preferably performed using a rotary vacuum concentrator, but is not limited thereto. The drying in the step 3) may be performed using hot air drying, vacuum drying or freeze drying, but is not limited thereto.

The extract of the extract of the alfalfa solids obtained in the above manner, preferably the ethanol extract of the alfalfa solanaceae, more preferably the alcohol extract of the alfalfa solanaceae, is further fractionated to obtain the fraction of the Solanum tuberculosis extract . The 'fraction of Solanum tuberculosis extract' can be obtained by various fractionation methods known in the art. Preferably, the fraction is not limited thereto, but may be obtained by suspending an extract of alfalfa solubles in a water, preferably a mixed solution of water and alcohol, and then successively fractionating with a fraction solvent having a different polarity. The fraction solvent may be n-hexane, dichloromethane, ethyl acetate, and n-butanol. That is, the n-hexane fraction is a fraction obtained by fractionating n-hexane to the extract of Solanum tuberculosis. The dichloromethane fraction is fractionated by n-hexane and dichloromethane is added to the remaining water layer (water-soluble fraction) Wherein the ethyl acetate fraction is a fraction obtained by fractionating the remaining water layer (water-soluble fraction) with dichloromethane by adding ethyl acetate, and the n-butanol fraction is fractionated with ethyl acetate And fractionating the remaining water layer (water-soluble fraction) by adding n-butanol.

The extract or the fraction obtained from the extract of the Solanaceae mackerel are effective for the treatment or prevention of inflammatory diseases. Particularly, the extract or the fractions fractionated from the extract of the above-mentioned Solanum tuberculosis extract (1) can be used for (1) extracellular-regulated protein kinase (ERK), jNK (c-Jun NH 2 -protein kinase) (See FIG. 13), and (2) inhibiting the activation of IκB-α, which influences the expression of NF-κB (see FIG. 12), as well as the activity of NF-κB (See Figs. 10 and 11). Inhibition of NF-κB activity as described above inhibits the expression of iNOS and COX-2 (see FIG. 9) and the production of inflammatory cytokines (TNF-α, IL-1β and IL-6) Inhibited iNOS and COX-2 inhibited the production of nitrogen monoxide (NO) (see FIGS. 5 and 7) and inhibited the production of prostaglandin E ₂ (prostaglandin E ₂ , PGE ₂ ) (See Fig. 8 (a)). Through the above-described mechanism, the extract of the Solanaceae moth or the fraction extracted from the extract has an effect on the treatment or prevention of an inflammatory disease. In addition, the extract of the Allium cepa L. or the fraction obtained from the extract does not exhibit cytotoxicity even at a sufficiently high concentration (see FIGS. 4 and 6), and can be safely used for pharmaceutical use. In particular, it was confirmed that the size (thickness) of the inflammatory edema was reduced in the experimental group treated with the extract or the fraction extracted from the extract of the alfalfa solanaceae in the inflammatory edema-induced model animals (see Table 1) It was confirmed that the extract or extract of the extract of the present invention exhibited the therapeutic or preventive effect of the inflammatory diseases described above in vivo.

The pharmaceutical composition may contain at least one active ingredient which exhibits the same or similar function to the extract of the alfalfa solanaceae or the fraction fractioned from the extract, and may contain at least one kind of an appropriate carrier commonly used in the production of a pharmaceutical composition , Excipients, and diluents. The carrier, excipient and diluent may be selected from the group consisting of lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, Starch, gum Arabic, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, undecipitated starch, starch, gum arabic, alginate, gelatin, calcium phosphate, calcium silicate, But are not limited to, microcrystalline cellulose, polyvinyl pyrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium magnesium stearate, stearate, mineral oil, and the like. When the pharmaceutical composition is formulated, diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, surfactants and the like may be used, but the present invention is not limited thereto.

The pharmaceutical composition may be administered orally, rectally, intravenously, intramuscularly, subcutaneously, intraperitoneally, or intracerebrally in various routes, preferably orally. The pharmaceutical composition may be formulated for parenteral administration such as an external preparation such as an ointment such as an ointment, granule, tablet, capsule, suspension, or ointment, a suppository and a sterile injection solution, And the like. The solid preparation for oral administration may be prepared by mixing at least one excipient such as calcium carbonate, sucrose, lactose or gelatin in the fraction, , Lubricants such as magnesium stearate, talc, etc. may also be used. Liquid preparations for oral administration may be prepared by mixing at least one diluent or excipient, for example, a wetting agent, a sweetening agent, a fragrance, a preservative, etc., in the fraction. The preparation for parenteral administration may be prepared by mixing a sterilized aqueous solution, a non-aqueous solvent, a suspending agent, an emulsion, a freeze-drying agent and the like. Examples of the non-aqueous solvent or suspension include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, and the like can be used. Particularly, when formulated as a suppository, witepsol, polyethylene glycol, tween 61, cacao paper, laurin, glycerogelatin and the like may be used as the base material .

Although the preferred dosage of the pharmaceutical composition varies depending on the age, condition, weight, disease severity, form of the drug, administration route and time of the patient, the dosage of the active ingredient for the generally preferred effect is 0.001 to 1 Kg / day, preferably 0.001 to 0.1 mg / kg, and may be administered several times a day, preferably one to six times, at regular intervals according to the judgment of a doctor or pharmacist.

2. A health functional food or cosmetic composition for preventing or ameliorating an inflammatory disease

Another aspect of the present invention is a method for treating Myagropsis myagroides extract, or a fraction of the above extract as an active ingredient. The present invention also provides a health functional food for preventing or ameliorating an inflammatory disease.

In addition, another aspect of the present invention is the use of Myagropsis myagroides extract or a fraction of the above extract as an active ingredient. The present invention also provides a cosmetic composition for preventing or ameliorating an inflammatory disease.

The extracts of the Solanum tuberculosis or the fractions fractionated from the extract are the same as those described in the item " 1. Pharmaceutical composition for the prophylaxis or treatment of inflammatory diseases ". Therefore, a detailed description thereof will be omitted because of the description of the item " 1. Pharmaceutical composition for the prevention or treatment of inflammatory disease & quot ;, and the following description will be given only of the composition specific to the health functional food and the cosmetic composition.

When the extract of the present invention or the fraction of the above extract is used as a food additive, the above extract or fraction can be used as it is or can be used together with other food or food ingredients and can be suitably used according to a conventional method . The amount of the active ingredient to be mixed can be suitably determined according to its intended use (prevention, health or therapeutic treatment). Generally, the composition of the present invention is added in an amount of not more than 15 parts by weight, preferably not more than 10 parts by weight, based on the raw material, when the food or beverage is produced. However, in the case of long-term consumption intended for health and hygiene purposes or for health control purposes, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range .

There is no particular limitation on the kind of the food. Examples of the food to which the above substances can be added include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include healthy 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 g to 0.04 g, preferably about 0.02 g to 0.03 g per 100 ml of the composition of the present invention.

In addition to the above, the extract of the present invention or a fraction thereof may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusting agents, stabilizers, Alcohols, carbonating agents used in carbonated drinks, and the like. In addition, the extract or fraction of the present invention may contain flesh for the production of natural fruit juice, fruit juice beverage and vegetable beverage. These components may be used independently or in combination. The ratio 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 extract or fraction of the present invention.

The cosmetic product prepared by containing the extract of the extract of the present invention or the fraction of the extract as an active ingredient of the present invention can be prepared in the form of a general emulsified formulation and a 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 can also be prepared in the form of a foam or an aerosol composition further containing a compressed propellant.

In addition, the cosmetic composition may contain, in addition to the extract of the present invention or its fractions, a lipid, an organic solvent, a solubilizing agent, a thickening agent and a gelling agent, a softening agent, an antioxidant, a suspending agent, a stabilizer, a foaming agent, , Water, ionic or nonionic emulsifiers, fillers, sequestering agents and chelating agents, preservatives, vitamins, barrier agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or cosmetics And may contain adjuvants conventionally used in the cosmetics field, such as any other ingredients used.

Hereinafter, the present invention will be described in detail with reference to examples and preparation examples.

It should be noted, however, that the following examples and preparative examples are provided only for the understanding of the present invention, and the present invention is not limited by the following examples and preparative examples.

An alien dog ( Myagropsis myagroides ) Preparation of extract

Figure 1 in the same way as in (a), a Solitary collected in Busan Gijang County and Jeollanamdo Wando County dog Sargassum (Myagropsis myagroides ). The details are as follows.

<1-1> Preparation of methanol extract

Myagropsis myagroides were naturally dried in sunshine, pulverized and powdered. 2 kg of the dried powder of the alfalfa solubles were each immersed in 10 L of 100% methanol, stirred well, and then subjected to reflux cooling and extraction three times at 70 DEG C for 1 hour to 5 hours, followed by filtration with a filter paper (Watts, Filtered. The filtrate thus obtained under reduced pressure was concentrated under reduced pressure using a rotary rotary evaporator at 40 ° C to remove the solvent, and the remaining residue was extracted as an extract of Solanum tuberculosis.

As a result, 365.5 g of a methanol extract of Solanum tuberculosis was obtained.

<1-2> Preparation of ethanol extract

The extract was prepared in the same manner as in Example <1-1>, except that 100% ethanol instead of 100% methanol was used as an extraction solvent.

As a result, 364.2 g of an ethanol extract of Solanum tuberculosis was obtained.

<1-3> Manufacture of alcohol extracts

The extract was prepared in the same manner as in Example <1-1>, except that a fermentation alcohol (Korean WOO Co., Ltd., Korea) having an ethanol concentration of 105% instead of 100% methanol was used as an extraction solvent.

As a result, 358.8 g of the alcohol extract of Alaska pollack was obtained.

<1-4> Preparation of acetone extract

The extract was prepared in the same manner as in Example <1-1>, except that 70% acetone instead of 100% methanol was used as an extraction solvent.

As a result, 372.3 g of acetone extract of Solanum tuberculosis was obtained.

Uncle Myagropsis myagroides ) Fractionation of Extracts

1 (b), fractions were obtained from the extract of Myagropsis myagroides obtained in Example 1 above. The details are as follows.

<2-1> Preparation of n-hexane fractions

4 L of a mixed solvent in which n-hexane, alcohol and water were mixed at a ratio of 10: 9: 1 was added to 100 g of the extract of the alcoholic beverages produced in the above Example <1-3> The water layer and the n-hexane layer were allowed to separate. After sufficient time had passed, the mixture was separated into 2 L water layer and 2 L n-hexane layer. 2 L of n-hexane was added again to the 2 L water layer thus separated, and the process of separating 2 L of n-hexane layer was repeated 3 times. As a result, a total of 8 L of n-hexane layer and 2 L of water layer were finally obtained.

The thus-obtained 8 L n-hexane layer was concentrated under reduced pressure at 40 DEG C by vacuum rotary evaporator to remove the solvent, and the remaining residue 151.1 g was obtained as a n-hexane fraction.

<2-2> Preparation of dichloromethane fractions

2 L of dichloromethane was added to the 2 L water layer finally obtained in Example <2-1>, and the process of separating 2 L of the dichloromethane layer was repeated 3 times. As a result, a total of 6 L of a dichloromethane layer and 2 L of a water layer were finally obtained.

The 6 L dichloromethane layer thus separated was concentrated under reduced pressure at 40 캜 by vacuum rotary evaporator to remove the solvent, and 0.8 g of the remaining residue was obtained as a dichloromethane fraction.

<2-3> Preparation of Ethyl Acetate Fractions

2 liters of ethyl acetate was added to 2 liters of the water layer finally obtained in Example <2-2>, and 2 liters of the ethyl acetate layer was separated. The procedure was repeated three times. As a result, a total of 6 L of ethyl acetate layer and 2 L of water layer were finally obtained.

The 6-L ethyl acetate layer thus separated was concentrated under reduced pressure at 40 ° C by vacuum rotary evaporator to remove the solvent, and 0.8 g of the residue was obtained as an ethyl acetate fraction.

<2-4> Preparation of n-butanol fractions

2 L of n-butanol was added to the final 2 L water layer obtained in Example <2-3>, and the process of separating 2 L of the n-butanol layer was repeated three times Respectively. As a result, a total of 6 L of n-butanol layer and 2 L of water layer were finally obtained.

The 6 L of the n-butanol layer separated as described above was concentrated under reduced pressure at 40 캜 by a vacuum rotary condenser to remove the solvent, and 0.5 g of the remaining residue was obtained as the n-butanol fraction.

Component analysis

<3-1> Determination of content of polyphenolic compounds

A portion of the methanol, ethanol, alcohol and acetone extracts of the alfalfa solids obtained in each of Examples <1-1> to <1-4> was first reacted with 1N of Folin-Ciocalteu reagent Next, the contents of the polyphenolic compounds contained in the four kinds of extracts of the alopecia areata were determined by reacting with 20% sodium carbonate. Phloroglucinol was used as a reference material.

As a result, as shown in Fig. 2, the amount of the polyphenolic compound in the acetone extract of the Solanaceae mackerel was only slightly higher than that in the other three Solanaceae matsumana extracts. (Fig. 2).

<3-2> Determination of the content of fucoxanthin

To confirm the contents of the fucoxanthin components contained in the extracts of Solanum tuberosum and Solanum tuberosus nucifera obtained in Examples <1-3> and <2-1>, Shimadzu HPLC system (Kyoto, Japan) were subjected to high performance liquid chromatography (HPLC). The Shimadzu HPLC system (Kyoto, Japan) was equipped with a pump (Shimadzu LC-20AD), a photodiode array detector (Shimadzu SPD-M20A), an automatic sample injector (SIL-20A), a system controller (CBM- Shimadzu LCsolution (ver.1.22sp) data analysis program.

For the analysis of the content of fucoxanthin, the extracts of the extracts of Solanum tuberculosis and Solanum tuberospora nucifera obtained in the above Examples <1-3> and <2-1> were respectively dissolved in 100 ppm And 2 microliters were separated by HPLC on a Luna RP-18 column (Luna C18 (2), 3 μm, 150 x 3.0 mm, Phenomenex, Canada). As the separation solvent, 100% methanol (solvent A) and 0.1% aqueous solution of formic acid (solvent B) were used. First, the mixing ratio of the solvent A / B was set at a ratio of 95: 5 at a ratio of 95: 5 for 100 minutes, then flowed at a ratio of 95: 5 for 10 minutes, 0.0 &gt;% &lt; / RTI &gt; solvent.

As a result, as shown in Fig. 3 (a), fucosanthin as a reference material was separated at about 46 minutes, while the extracts of Solanum tuberosum and Solanum tuberosus as shown in Figs. 3 (b) No peaks corresponding to fucosanthin were observed in the n-hexane fractions of canola seedlings at about 46 minutes.

From the above results, it can be seen that fucoxanthin is not present in the extracts of Solanum tuberosum and Solanum tuberosum obtained in Examples <1-3> and <2-1>, respectively. have.

An alien dog ( Myagropsis myagroides ) Cytotoxic effect of extracts and nitrogen monoxide ( NO ) Production inhibitory effect

<4-1> Treatment of Solanum tuberculosis extract

Mouse macrophage RAW 264.7, obtained from the American Type Culture Collection (ATCC, Rockville, MD, USA) was inoculated into DMEM supplemented with 10% fetal bovine serum (FBS), 100 units / ml penicillin, 100 ug / ml streptomycin And cultured in medium at 37 ° C and 5% CO ₂ .

The RAW 264.7 cells cultured as described above were dispensed in a 20-well plate at a concentration of 5 × 10 ⁴ cells / well or 1 × 10 ⁵ cells / well and then cultured in different concentrations (0, 25, 50, 100 μg / Were cultured for 1 hour in DMEM medium containing the respective samples (methanol, ethanol, alcohol and acetone extracts of the alfalfa mushroom obtained in each of Examples <1-1> to <1-4> above) Gt; / ml &lt; / RTI &gt; of LPS and cultured for 24 hours.

<4-2> Cytotoxic effect of Extracts from Solanaceae moth

To confirm the safety of methanol, ethanol, alcohol and acetone extracts of the mothballs obtained in each of Examples <1-1> to <1-4>, cytotoxicity tests by MTS analysis were conducted.

As in Example <4-1>, 100 μl of the medium was taken from RAW 264.7 cell cultures treated with Solanum tuberculosis extracts, and each well was dispensed into a 96-well plate together with 5 μl of MTS solution and reacted for 1 hour , And the absorbance was measured at 490 nm using a microplate reader. Measurements were determined as the mean value of three replicate experiments, and relative cell viability (% of control) was calculated by comparing the determined values with the control group.

As a result, as shown in FIG. 4, no methanol, ethanol, alcohol, or acetone extract of the Allium tuberculosis showed cytotoxicity up to a concentration of 100 μg / ml (FIG. 4).

<4-3> Inhibitory Effect of Extracts from Allium cepa L. on the Formation of Nitric Oxide (NO)

Green LC, etc. (Anal Biochem. (1982)) was used to examine the inhibitory effect of methanol, ethanol, alcohol, and acetone extract on the production of nitrogen monoxide in the alfalfa solids obtained in each of Examples <1-1> to < 126: 131-138). &Lt; tb &gt; &lt; TABLE &gt;

As in Example <4-1>, 100 .mu.l of the medium was divided into 96-well plates in RAW 264.7 cell cultures treated with Solanum tuberculosis extracts, and 100 .mu.l of Griess reagent was added to each well After incubation for 10 minutes, the absorbance was measured at 540 nm using an ELISA reader. Standard concentration curves were obtained by serial dilution of sodium nitrite (NaNO ₂ ).

As a result, as shown in FIG. 5, the production of LPS-induced nitric oxide in RAW 264.7 cells was decreased in a concentration-dependent manner by methanol, ethanol, The methanol extract, ethanol, alcohol, and acetone extract showed the highest inhibitory effect on nitrogen monoxide production (Fig. 5).

Uncle Myagropsis myagroides ) Cytotoxic effect and inhibitory effect on nitrogen monoxide (NO) production of extract fraction

<5-1> Treatment of Fractions of Solanum tuberculosis Extract

Mouse macrophage RAW 264.7, obtained from the American Type Culture Collection (ATCC, Rockville, MD, USA) was inoculated into DMEM supplemented with 10% fetal bovine serum (FBS), 100 units / ml penicillin, 100 ug / ml streptomycin And cultured in medium at 37 ° C and 5% CO ₂ .

The RAW 264.7 cells cultured as described above were dispensed in a 20-well plate at a concentration of 5 × 10 ⁴ cells / well or 1 × 10 ⁵ cells / well and then cultured in different concentrations (0, 25, 50, 100 μg / (Fraction of alcohol extract obtained from Example <1-3> and fraction of alcohol extract of alcohol extract obtained from each of Examples <2-1> to <2-3> obtained in Example <1-3> Ml DMEM medium for 1 hour, followed by treatment with 1 μg / ml of LPS and culturing for 24 hours.

<5-2> Cytotoxic effect of fractions of extracts

To confirm the safety of the fractions obtained from the extracts obtained in Examples <2-1> to <2-3>, the cytotoxicity test by MTS analysis was carried out.

As in Example <5-1>, 100 μl of the medium was taken from the RAW 264.7 cell cultures treated with the extracts of Solanum tuberculosis extract, and the cells were subcultured in a 96-well plate together with 5 μl of MTS solution, , And the absorbance was measured at 490 nm using a microplate reader. Measurements were determined as the mean value of three replicate experiments, and relative cell viability (% of control) was calculated by comparing the determined values with the control group.

As a result, as shown in FIG. 6, the n-hexane, dichloromethane or ethyl acetate fractions of the Solanum tuberculosis extract showed no cytotoxicity up to a concentration of 100 μg / ml (FIG. 6).

<5-3> Inhibitory Effect of Fraction of Extracts from Extracts of Allium tuberosum L. on the Formation of Nitric Oxide (NO)

Green LC et al. (Anal Biochem. (1982) 126: 131-138) were used to confirm the inhibitory effect on the production of nitrogen monoxide by the fractions of the extracts of Solanum tuberculosis extract obtained in each of Examples <2-1> to < Experiments were carried out in accordance with the method for measuring the amount of nitrogen monoxide produced in the literature.

As in Example <5-1>, 100 .mu.l of the culture medium was divided into 96-well plates in RAW 264.7 cell cultures treated with Solanum tuberculosis extracts, 100 .mu.l of Griess reagent was treated in each well After incubation for 10 minutes, the absorbance was measured at 540 nm using an ELISA reader. Standard concentration curves were obtained by serial dilution of sodium nitrite (NaNO ₂ ).

As a result, as shown in Fig. 7, the production of LPS-induced nitric oxide in RAW 264.7 cells was decreased in a concentration-dependent manner by n-hexane, dichloromethane or ethyl acetate fractions of Solanum tuberculosis extract, Among the n-hexane, dichloromethane and ethyl acetate fractions of the extracts of N. chinensis, the n-hexane fraction of the extracts of the extracts of the Euphorbiaceae showed the highest inhibitory effect on the production of nitrogen monoxide (FIG. 7).

An alien dog ( Myagropsis myagroides ) Alcohol extract of the alcohol and alcohol extract of the alcohol Hexane  Phosphorus of fraction In vitro  Identification of inflammation inhibitory effect

<6-1> Treatment of n-hexane fractions of alcohol extracts from alcoholic beverages and alcohol extracts

Mouse macrophage RAW 264.7, obtained from the American Type Culture Collection (ATCC, Rockville, MD, USA) was inoculated into DMEM supplemented with 10% fetal bovine serum (FBS), 100 units / ml penicillin, 100 ug / ml streptomycin And cultured in medium at 37 ° C and 5% CO ₂ .

The RAW 264.7 cells cultured as described above were dispensed into a 24-well plate at a concentration of 1 × 10 ⁵ cells / well, and samples of different concentrations (0, 25, 50, and 100 μg / ml) 1-3>, and the n-hexane fraction of the alcohol extract of the Allium tuberosum extract obtained in Example <2-1>) for 1 hour, and then cultured in DMEM medium containing 1 Gt; / ml &lt; / RTI &gt; of LPS and cultured for 24 hours.

<6-2> Confirming the inhibitory effect of inflammatory cytokines and chemokines

In order to examine the inhibitory effect of the n-hexane fraction of the alcohol extract obtained from Example <1-3> and the alcohol extract obtained from Example <2-1> in Example _{1 to} inhibit PEG ₂ formation Respectively.

100 [mu] l of the medium was taken from the RAW 264.7 cell culture treated with Solanum tuberculosis extract or fraction as in Example <6-1>, and the inflammatory cytokine secreted in the medium by ELISA kit (BD Biosciences, USA) cytokine) and chemokines (chemokine) of prostaglandin _{E 2 (prostaglandin E 2, PGE} 2), TNF (tumor necrosis factor-α, TNF -α), interleukin--1β (interleukin-1β, IL- 1β) and interleukin- 6 (interleukin-6, IL-6) were measured.

As a result, the production of LPS-induced PEG ₂ , TNF-α, IL-1β and IL-6 in RAW 264.7 cells, as shown in FIGS. 8A to 8D, (Fig. 8), and the n-hexane fraction of the extracts of the extracts and the extracts of Allium tuberospores (Fig. 8).

<6-3> Identification of inhibition of protein production by iNOS and COX-2

To investigate whether the inhibition of nitric oxide (NO) and prostaglandin E ₂ (prostaglandin E ₂ , PGE ₂ ) production by inhibition of iNOS and COX-2 expression was inhibited by iNOS and COX-2 proteins The effect of the n-hexane fraction of the alcohol extracts and alcohol extracts from the alfalfa extract was confirmed by western blot and RT-PCR.

As a result, as shown in Figs. 9 (a) and 9 (b), the expression of LPS-induced iNOS in RAW 264.7 cells was inhibited by extracting alcohol extracts from Allium cepa . The expression of COX-2 was significantly decreased in the n-hexane fraction of the extracts of the extracts of the Allium miltiorrhizae extract and the Allium sativum extract of the alopecia mellifera at a concentration of 100 μg / ml (FIG. 9).

From the above results, it was confirmed that the extracts of the extracts of the extracts of the alfalfa solanaceousum and the alfalfa solanaceae inhibited the production of nitrogen monoxide (NO) by inhibiting the production of iNOS by affecting the protein expression of iNOS .

On the other hand, the n-hexane fractions of the extracts of the extracts of the extracts of the Solanaceae mushroom and Solanaceae mushroom extract inhibit the protein expression of COX-2 only at a relatively high concentration. This is because the n-hexane fraction of the alcohol extract and the alcohol extract of the alfalfa extract of the alfalfa sprout affect the activation of COX-2 protein rather than the expression of COX-2 protein.

<6-4> Confirming the inhibitory effect of NF-κB activation

Inhibition of NF-κB expression by iNOS, COX-2, TNF-α, IL-1β and IL-6 The effect of the n-hexane fraction of the extract from the ganoderma lucidum extract was confirmed by luciferase assay, immunostain and western blot.

First, in the luciferase assay, a recombinant gene in which luciferase was recombined in the promoter site of NF-κB was introduced into RAW 264.7 cells cultured as in Example <6-1> and stabilized , Treating the stabilized cells with the ethanol extract of the alfalfa sprouts and the ethanol extract of the alfalfa sprouts, and measuring the luminescence from the cell lysate. As a result, as shown in Fig. 10, the activation of LPS-induced NF-κB transcription factor in RAW 264.7 cells was inhibited by the n-hexane fraction of the alcohol extract of Solanaceae moth (Fig. 10).

Next, in order to examine the influence of the n-hexane fraction of the alcohol extract of the Allium tuberculosis and the alcohol extract of Allium tuberculosis, when the NF-κB was transferred to the nucleus, as shown in Example <6-1> % fetal bovine serum (FBS), 100 units / ㎖ penicillin, 100 ㎍ / ㎖ streptomycin one to 37 ° C, the sample with 5% by RAW 264.7 cells cultured in a CO ₂ environment in a DMEM medium was added (example <1 -3 > and the n-hexane fraction of the alcohol extract of Allium tuberosum obtained in Example < 2-1 &gt;) for 1 hour, and then cultured in DMEM medium containing 1 ㎍ / Ml of LPS and cultured for 1 hour. RAW 264.7 cells cultured as described above were immunostained with NF-κB antibody and DAPI, and then the intracellular location of NF-κB was observed using a confocal microscope. As a result, as shown in Fig. 11, the migration of LPS-induced NF-κB to the nucleus in RAW 264.7 cells was inhibited by the n-hexane fraction of the alcohol extract of the alfalfa moth (Fig. 11).

Finally, in order to examine the influence of the n-hexane fraction of the alcohol extract of Allium tuberculosis and the alcohol extract of Allium tuberculosis, on the phosphorylation of IκB-α and the migration of NF-κB to the nucleus,

The cells were cultured in DMEM medium supplemented with 10% fetal bovine serum (FBS), 100 units / ml penicillin and 100 μg / ml streptomycin at 37 ° C and 5% CO ₂ as in Example <6-1> RAW 264.7 cells were inoculated with DMEM containing the samples (the alcohol extract obtained from the above Example <1-3> and the n-hexane fraction of the alcohol extract obtained from Example <2-1>) After culturing for 1 hour in the medium, 1 ㎍ / ml of LPS was treated and cultured for 30 minutes. Cell lysate was recovered from RAW 264.7 cells cultured as described above, and western blotting was performed using pIκB-α, IκB-α and NF-κB antibodies. As a result, as shown in FIG. 12, the phosphorylation of IκB-α in the cytoplasm and the migration of NF-κB into the nucleus and the nuclear transfer of nuclear The amount of NF-κB was inhibited in a concentration-dependent manner (FIG. 12).

From the above results, it was found that the n-hexane fraction of the alcohol extract of the Allium cepa L. and the alcohol extract of Allium tuberosporium alfalfa inhibited phosphorylation and degradation of IκB-α induced by LPS in a concentration-dependent manner, And to the nucleus, blocking the function of NF-kB as a transcription factor.

<6-5> Identification of inhibitory effects of MAPKs / Akt pathway

Extracted extracts of Solanum tuberosum and Solanum tuberculosis mellifera against extracellular-regulated protein kinase (ERK), JNK (c-Jun NH2-protein kinase) and p38 MAPK and PI3K / Akt activated by transcription factor activity of NF- The effect of the n-hexane fraction of the extract was confirmed by western blotting.

As a result, as shown in FIG. 13, the n-hexane fractions of the extracts of the alcohol extracts of Solanaceae mushroom and Solanaceae mushroom extract of the present invention showed phosphorylation of ERK, JNK and p38 MAOK induced by RAW 264.7 cells And phosphorylation of Akt downstream of PI3K in a concentration-dependent manner.

An alien dog ( Myagropsis myagroides ) Alcohol extract of the alcohol and alcohol extract of the alcohol Hexane  The in vivo fraction of the fraction in vivo ) On inflammation inhibition

<7-1> Breeding of experimental animals

An 8-week-old ICR mouse (Korea BioLink Co., Ltd.), which is the most used animal model for inflammatory animal models, was cultured at 20 to 22 캜 and 50% to 60% , And normal animals were selected by observing gross symptoms and randomly distributed in groups of 10 mice per group.

<7-2> Breeding of experimental animals

The extracts of the extracts of Alfalfa solanaceae and the extracts of Alfalfa solanaceae were applied to the right ear of the experimental animals as described in Example <7-1>. After 1 hour, 6 占 퐂 of phorbol 12-myristate 13-acetate (PMA) was dissolved in 30 占 퐇 of acetone solution and then applied to the right ear and the left ear of the experimental animal. Six hours after application, animals were anesthetized and the thickness of each ear was measured in micrometers to calculate the rate of increase as an index of edema. The 100% edema index was defined as the group treated only with MPA and the relative edema rate (% of control) was suggested. All measurements were expressed as means and standard deviations. Statistical analysis was performed using Student's t-test to determine differences between the groups. Statistical significance was determined for p <0.05 values.

As a result, 66.8% of the group treated with the extract of alcoholic beverages and 44.8% of the group treated with the n-hexane fraction of alcoholic beverages showed the suppression of edema compared to the control group treated with PMA alone One).

Control group Extract treatment group Fraction treatment group PMA ○ ○ ○  The extract of Example <1-3> × ○ × The fraction of Example <2-1> × × ○ Suppression of edema (%) 100 66.8 44.8

Examples of formulations for the composition of the present invention are illustrated below.

&Lt; Formulation Example 1 > Preparation of pharmaceutical preparation

The pharmaceutical preparations comprising the extracts of Solanum tuberculosis of the present invention or fractions thereof are prepared as follows.

1. Manufacturing of powder

The extract of Example 1 or the fraction of Example 2

Lactose 1g

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

2. Preparation of tablets

100 mg of the extract of Example 1 or the fraction of Example 2

100 mg of corn starch

100 mg of milk

2 mg of magnesium stearate

After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.

3. Preparation of capsules

100 mg of the extract of Example 1 or the fraction of Example 2

100 mg of corn starch

100 mg of milk

2 mg of magnesium stearate

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

4. Manufacture of rings

The extract of Example 1 or the fraction of Example 2 1 g

Lactose 1.5 g

Glycerin 1 g

0.5 g of xylitol

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

5. Manufacture of granules

150 mg of the extract of Example 1 or the fraction of Example 2

Soybean extract 50 mg

200 mg of glucose

600 mg of starch

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

&Lt; Formulation Example 2 > Production of food

Foods containing the extracts of the present invention or fractions thereof are prepared as follows.

1. Manufacture of Flour Food

0.5 to 5.0 parts by weight of the extract of Example 1 of the present invention or the fraction of Example 2 were added to wheat flour and the mixture was used to prepare breads, cakes, cookies, crackers and noodles, .

2. Manufacture of dairy products

5 to 10% by weight of the extract of Example 1 of the present invention or the fraction of Example 2 is added to milk and various dairy products such as butter and ice cream are prepared using the milk.

3. Manufacturing of wire

Brown rice, barley, glutinous rice, and yulmu are dried by a known method and dried, and then the mixture is ground to a powder having a particle size of 70 mesh. Black beans, black sesame, and perilla are steamed and dried by a known method, and then powdered with a powder having a particle size of 70 mesh by a grinder. The extract of Example 1 of the present invention or the fraction of Example 2 is concentrated under reduced pressure in a vacuum concentrator and dried by spraying and drying in a hot air drier to obtain a dried powder by pulverizing the dried material to a particle size of 70 mesh with a pulverizer. The grains, seeds and the dry powder of the extract of Example 1 or the fraction of Example 2 prepared above are blended in the following proportions.

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

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

The dried powder (3% by weight) of the extract of Example 1 or the fraction of Example 2,

(0.5% by weight),

(0.5% by weight)

&Lt; Formulation Example 3 > Preparation of beverage

1. Manufacture of health drinks

1000 mg of the extract of Example 1 or the fraction of Example 2

Citric acid 1000 mg

100 g of oligosaccharide

Plum concentrate 2 g

Taurine 2 g

Purified water was added to a total of 900 ml

The above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 DEG C for about 1 hour. The solution thus prepared was filtered and sterilized in a sterilized 2 liter container, It is used in the production of the health beverage composition of the invention.

The composition ratio is a mixture of the components suitable for the preferred beverage as a preferred embodiment. However, the blending ratio may be arbitrarily varied depending on the regional and national preference such as the demand level, the demanding country, and the intended use.

2. Manufacture of vegetable juice

5 g of the extract of Example 1 of the present invention or the fraction of Example 2 is added to 1,000 ml of tomato or carrot juice to prepare vegetable juice for health promotion.

3. Manufacture of fruit juice

Healthy fruit juice is prepared by adding 1 g of the extract of Example 1 of the present invention or the fraction of Example 2 to 1,000 ml of apple or grape juice.

&Lt; Formulation Example 4 > Preparation of cosmetics

A cosmetic product for preventing or ameliorating edema or various inflammation, which contains the extract of Solanum tuberculosis of the present invention or a fraction thereof as an active ingredient, can be produced. As the above-mentioned cosmetics, cosmetics of emulsified form such as nutrition lotion, cream, essence and the like, and cosmetics of solubilized form such as softened longevity can be produced

<4-1> Cosmetics manufacturing of emulsified formulations

An emulsifier-type cosmetic is produced in the composition shown in Table 1. The production method is as follows.

1) The mixture of ingredients 1 to 9 is heated to 65 to 70 占 폚.

2) The raw material of component 10 is added to the mixture of step 1).

3) Mixture of raw materials of components 11 to 13 is completely dissolved by heating to 65 to 70 캜.

4) While the above step 3) is carried out, the mixture of step 2) is gradually added and emulsified at 8,000 rpm for 2 minutes to 3 minutes.

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

6) Each of the raw materials of components 15 to 17 is weighed, then put into the mixture of step 5), and further emulsified for 30 seconds.

7) The mixture of step 6) is emulsified and then degassed and cooled to 25 ° C to 35 ° C to prepare an emulsifier-type cosmetic product.

Furtherance Emulsifier type 1 Emulsifier type 2 Emulsifier type 3 One Stearic acid 0.3 0.3 0.3 2 Stearyl 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 Cetyl ethyl hexanoate 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 The extract of Example 1
Or < Example 2 > fraction 0.0001 One 10

&Lt; 4-2 > Cosmetic preparation of solubilized formulation

A cosmetic product of the solubilized formulation is prepared with the composition shown in Table 2. The production method is as follows.

1) The raw materials of the components 2 to 6 are put into a raw material (purified water) of 1 and dissolved by using a mixer.

2) The ingredients 8 to 11 are completely dissolved in the raw material (alcohol) of 7.

3) The mixture of step 2) is slowly solubilized by adding it to the mixture of step 1).

Furtherance Solubilization Formulation 1 Solubilization Formulation 2 Solubilization 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 The extract of Example 1
Or < Example 2 > fraction 0.1 5 5 7 Alcohol (95%) 8 8 8 8 Methyl paraoxybenzoate 0.1 0.1 0.1 9 Polyoxyethylene
Hydro genide ester 0.3 0.3 0.3 10 incense 0.15 0.15 0.15 11 Cyclomethicone - - 0.2

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the scope of the invention is not limited to the disclosed exemplary embodiments. It will be possible to change it appropriately.

Claims (11)

delete delete delete delete delete A pharmaceutical composition for the prophylaxis or treatment of inflammatory diseases containing an n-hexane fraction of an extract of Myagropsis myagroides which does not contain fucoxanthin as an active ingredient. delete delete The method of claim 6, wherein the inflammatory disease is selected from the group consisting of dermatitis, allergy, atopy, asthma, conjunctivitis, periodontitis, rhinitis, otitis, iritis, sore throat, tonsillitis, pneumonia, gastric ulcer, pancreatitis, gastritis, Crohn's disease, Sjogren's syndrome and multiple sclerosis, including but not limited to rheumatoid arthritis, rheumatic fever, lupus, fibromyalgia, psoriatic arthritis, osteoarthritis, rheumatoid arthritis, shoulder periarthritis, tendinitis, Wherein the compound is selected from the group consisting of: &lt; RTI ID = 0.0 &gt; (I) &lt; / RTI &gt; A health functional food for preventing or ameliorating an inflammatory disease containing an n-hexane fraction as an active ingredient of an alcohol extract of alcoholic beverages containing no fucoxanthin. A cosmetic composition for preventing or ameliorating an inflammatory disease containing an n-hexane fraction of an alcohol extract of an Alaska pollack containing no fucoxanthin as an active ingredient.

Images