KR101879685B1 - Composition for anti-inflammation with nipa fruticans wurmb - Google Patents

Abstract

The present invention relates to a composition for antiinflammation using a nipponophyllum phalaenopsis, and more particularly, to a composition comprising a nipponophyllum phloem extract extracted from water, ethanol or a mixed solvent thereof as an effective ingredient and characterized by being a pharmaceutical composition .
The anti-inflammatory composition using the nifa palm phlox of the present invention has an effect of inhibiting or inhibiting the expression of an inflammatory factor and treating or alleviating inflammation.

Description

TECHNICAL FIELD [0001] The present invention relates to a composition for anti-inflammation using a nifa palm phalanx,

The present invention relates to a composition for antiinflammation using a nifa palm phlox, and more particularly, to a composition for anti-inflammation using a nifa palm phlox which has an excellent anti-inflammatory ability and contains no cytotoxicity Lt; / RTI >

Inflammation is the expression of a normal and protective in vivo defense mechanism that is localized to physical trauma, harmful chemicals, infections by microorganisms, and tissue damage caused by stimuli in vivo metabolites. These inflammations are triggered by various chemical mediators produced from damaged tissues and migrating cells, and these chemical mediators are known to vary according to the type of inflammation process. In normal cases, the organism neutralizes or eliminates the cause of inflammation through the inflammation reaction, regenerates the upper tissue and regenerates the normal structure and function, but if not, the disease state such as chronic inflammation also proceeds. In addition, anti-inflammatory drugs are needed when the inflammation is triggered improperly by the autoimmune reaction such as pollutant such as pollen, asthma or rheumatoid arthritis. Although most inflammatory diseases can be observed in almost all clinical diseases, some of these inflammatory diseases are bacterial diseases that can be cured by administration of antibiotics, but most of them are due to tissue damage due to autoimmune reaction, so there is no specific treatment It is known as an incurable disease.

The most common anti-inflammatory drugs for treating these inflammatory diseases are classified into steroidal and non-steroidal anti-inflammatory drugs. Most of the synthetic anti-inflammatory drugs have many side effects besides the main action, so they are excellent. The development of such a system is urgently required. In other words, use nonsteroidal anti-inflammatory drugs (NSAIDs) that are suitable for the treatment of inflammatory diseases to relieve inflammation, use steroids if inflammation is severe or NSAIDs are not effective, . In this case, the NSAIDs used in the present invention mainly inhibit cyclooxygenase (COX) and inhibit the production of prostaglandins which are involved in the inflammatory reaction, thereby exhibiting anti-inflammatory effects. However, the NSAIDs may cause gastrointestinal disorders, hepatic disorders, It causes side effects, making long-term use difficult. Compared to these NSAIDs, steroids are the fastest method available to patients and are known to be a rapidly and dramatically anti-inflammatory drug. However, as it is widely known, this drug weakens resistance to bacterial infections, causes deterioration of diabetes, adrenal insufficiency, and mental dysfunction. It is not only very toxic but also difficult to stop when starting treatment. It is known that it is necessary and should be banned if possible.

As described above, synthetic anti-inflammatory drugs are often accompanied by various side effects. Thus, there is a constant demand for the development of anti-inflammatory drugs which are effective but have relatively few side effects. Particularly, from the viewpoint of efficacy and side effects, natural herbal preparations, which are rich in clinical experience and excellent in safety, will be good candidates for the development of preventive and therapeutic agents for inflammatory diseases.

Conventionally, as a therapeutic agent for inflammation of a natural product preparation, Korean Patent No. 10-0991398 has been published, and further development of an effective inflammatory agent for a natural product preparation has been demanded.

KR 10-099139 B1

Accordingly, it is an object of the present invention to provide a pharmaceutical composition for anti-inflammation.

It is also intended to provide a health functional food composition for preventing or ameliorating inflammation.

In order to accomplish the above object, the present invention provides a composition for anti-inflammation using a nifa palm phlox, wherein the nifa palm phlox is extracted with water, ethanol, or a mixture of water and ethanol as a solvent, The flower bud extract is prepared by adding 3 to 20 times by weight dry weight of a solvent to a dried powder obtained by naturally drying and shredding nifa palm flower buds, Or a powder obtained by removing the solvent from the extract by hot water extraction or cold extraction or warm extraction or reflux cooling extraction or ultrasonic extraction for 30 minutes to 5 days, Maltitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, At least one of cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil as an active ingredient The extract of the present invention is a concentrate obtained by concentration under reduced pressure at 20 to 100 ° C. The concentrated extract or the powdered extract is used in water or ethanol or a mixture of water and ethanol, The hibiscus extract is a hibiscus extract obtained by naturally drying a flower of a hibiscus safflower paraffin, and drying the dried flower powder. , And the mixture was extracted at an extraction temperature of 20 to 100 ° C for 30 minutes to 5 days, The powder being removal of the solvent from the extract-containing further, the spine Nipa palm extract, hibiscus extract the 1: a 0.1 to 0.2 ratio by weight may contain, as an active ingredient.

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The anti-inflammatory composition using the nifa palm phlox of the present invention has an effect of inhibiting or inhibiting the expression of an inflammatory factor and treating or alleviating inflammation.

1 is a graph showing the results of a cytotoxicity test of Example 1 according to the present invention.
2 is a graph showing the results of the cytotoxicity test of Example 2 according to the present invention.
3 is a graph showing the results of a test for inhibiting nitrite generation in Example 1 according to the present invention.
4 is a graph showing the results of a test for inhibiting nitrite formation in Example 2 according to the present invention.
Fig. 5 is a graph showing the results of the IL-1? Inhibition assay of Example 1 of the present invention. Fig.
6 is a graph showing the results of a test for inhibiting IL-1β expression in Example 2 according to the present invention.
7 is a graph showing the results of the IL-6 expression inhibition activity test of Example 1 according to the present invention.
8 is a graph showing the results of a test for inhibiting the expression of IL-6 in Example 2 according to the present invention.
9 is a graph showing the results of the TNF-α expression inhibition activity test of Example 1 according to the present invention.
10 is a graph showing the results of a TNF-α expression inhibition activity test of Example 2 according to the present invention.

Hereinafter, the present invention will be described in detail.

The composition for anti-inflammation of the present invention contains an extract of the nipponophyllum phalaenopsis as an active ingredient.

The above-mentioned NIPA FRUTICANS WURMB is originated in India, Malaysia, Southeast Asia and Australia, and grows in wetlands such as mangroves. The roots are divided into several pieces in the ground, and the leaves are gathered on the ground and have a glossy green color. And the flower is one male and one female, and it comes out from the axilla of the ground. Usually, the flower stands rise from October to June of the following year.

The reason for using the flower bud of nipaya is that it has abundant antioxidant components such as polyphenols in the flower bud than any other part of nipaya, and the selenium And other nutritional ingredients, as well as show excellent anti-inflammatory effects.

In the present invention, the extract of the nifa palm phlox is extracted from the dried powder of the nifa palm phalaenopsis.

At this time, the dried powder of the nihon paladin can include not only a nippae flower, but also a flower bud or a flower attached to a flower bed, and the nipponia palm is picked, dried and crushed. Here, the drying may be carried out in a known manner to such an extent that the useful components of the harvested napa are not destroyed. For example, the drying may be carried out by natural drying in the shade. In addition, the crushing is sufficient for crushing the crust to such an extent that the useful components of the nipponophyllum peduncle can be sufficiently extracted in the subsequent extraction process. In addition, the drying and crushing processes may be carried out in reverse order or repeatedly as required.

The extraction may be, for example, hot water extraction, cold extraction, warming extraction, reflux cooling extraction, ultrasonic extraction, or the like.

In addition, the extraction is carried out using a solvent, and water, ethanol, or a mixed solvent thereof may be used as the solvent.

The extraction is carried out by adding 3 to 20 times by weight of the dry weight of the dried powder to the nifa palm phlox, and more preferably at 30 to 5 days at an extraction temperature of 20 to 100 ° C, more preferably 40 to 90 ° C A method of extracting for 1 to 24 hours can be applied.

At this time, the liquid extract may be separated from the dried powder of the nifalia phalaenopsis by a method such as filtration, followed by concentration or drying. For example, the extract of the liquid phase may be a concentrate obtained by concentrating the extract at a reduced pressure at 20 to 100 ° C, preferably 40 to 70 ° C by using a vacuum rotary condenser, and drying the extract to obtain a powdered powdery extract. The extracted or concentrated extract may be used in water, ethanol or a mixed solvent thereof if necessary.

These extracts of Nepharahia phalaenopsis are useful for the prevention and treatment of inflammation-related diseases by inhibiting the expression of inflammatory factors such as Nitrite, interleukin-1?, Interleukin-6 and TNF-a.

The antiinflammatory composition comprising the Isoproterenol extract of the present invention can be used as a pharmaceutical composition for inhibiting the expression of an inflammatory factor.

In addition, the anti-inflammatory composition may further comprise additional components, for example, at least one of a carrier, an excipient, and a diluent, and more specifically, lactose, dextrose, sucrose, But are not limited to, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, Hydroxybenzoate, hydroxybenzoate, talc, magnesium stearate, and mineral oil.

The anti-inflammatory composition may have various formulations and may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols or other external preparations according to a conventional method. In the case of formulation, it can be prepared using diluents or excipients such as fillers, extenders, binders, humectants, disintegrants, surfactants and the like which are usually used, and the preparations for non-oral administration include sterilized aqueous solutions, Emulsions, and freeze-dried preparations. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like.

The preferred dosage of the anti-inflammatory composition may be appropriately selected depending on the condition and body weight of the patient, the degree of disease, the form of the drug, the route of administration and the period of time. However, for the desired anti-inflammatory effect, the extract or the compound of the present invention can be administered in an amount of 0.01 to 500 mg / kg per day, preferably 0.1 to 200 mg / kg per day, divided once to several times per day. In addition, the dose of the compound may be increased or decreased depending on the route of administration, degree of disease, sex, weight, age, and the like.

In addition, the anti-inflammatory composition of the present invention can be used as a food composition, and has an anti-inflammatory function including an extract of Nippon Ranviaceae, and is used as a food composition in the form of a health functional food, a nutritional supplement, a health food, or a food additive .

Food compositions of this type can be prepared in a variety of forms according to conventional methods. For example, the health food may be prepared by liquefying, granulating, encapsulating and pulverizing the extract in the form of tea, juice and drink so that it can be consumed. In addition, it can be prepared in the form of a composition by mixing together the above-mentioned Nipah palm phloem extract and a known active ingredient known to have anti-inflammatory effect. Functional foods also include, but are not limited to, beverages (including alcoholic beverages), fruits and processed foods such as canned fruits, jars, jam, maare marlade, fish, meat and processed foods (Eg butter, cheeses, etc.), edible vegetable oils (eg, sourdoughs, etc.), breads and noodles (eg udon, buckwheat noodles, ramen noodles, spaghetti, macaroni, , Margarine, vegetable protein, retort food, frozen food, various kinds of seasonings (for example, soybean paste, soy sauce, sauce, etc.). The preferred content of the above-mentioned A. napaya phloem extract in the food composition is not particularly limited, but is preferably 0.01 to 50% by weight in the finally prepared food.

Meanwhile, the anti-inflammatory composition of the present invention may further contain, as an active ingredient, a hibiscus extract obtained by extracting flowers of Hibiscus subtilis paraine with water, ethanol or a mixed solvent thereof.

The extract of Hibiscus is known to have an excellent antibacterial effect. When the Hibiscus extract is applied as an anti-inflammatory composition together with the extract of the present invention, the anti-inflammatory effect, that is, the effect of suppressing the expression of the inflammatory factor is remarkably increased compared to the concentration.

Therefore, it is possible to obtain a more excellent anti-inflammatory effect by only a small amount of the extract of Nihon palm phalaenopsis and hibiscus extract.

The Hibiscus sabdariffa Linn (English name Roselle) is an edible hibiscus, and it is preferable that the flower extract is extracted in the same manner as the above-described Nepharahia phalaenopsis extract, so that a detailed description of this method is omitted do. That is, an extract obtained by adding a dry weight of 3 to 20 times by weight of a solvent to the dried powder of flower of Hibiscus safflower parin and extracting it at an extraction temperature of 20 to 100 ° C for 30 minutes to 5 days, The removed powder can be used. As the solvent, water, ethanol or a mixed solvent thereof may be used. Here, the dried powder of the flower of the Hibiscus safflower parin is also prepared by drying and crushing the flower of the collected hibiscus safflower parin. At this time, the useful components of the flowers of the Hibiscus subtilis paraffin can be dried in a known manner, for example, by natural drying in a shade field. In addition, it is sufficient that the crushing is carried out so that the useful components of the flowers of the Hibiscus subsp. Paraffin can be sufficiently extracted in the subsequent extraction process. In addition, the drying and crushing processes may be carried out in reverse order or repeatedly as required.

In addition, in the present invention, it is most preferable that the extract of the present invention contains the above-mentioned Nihon palm phalaenopsis extract and the above-mentioned Hibiscus extract as an active ingredient at a weight ratio of 1: 0.1-0.2, and exhibits the most excellent anti-inflammatory activity in this mixing ratio.

Hereinafter, the present invention will be described in more detail with reference to specific examples.

(Example 1): Preparation of Extract of Isolate Nymphalidae.

The flower buds of Nipaya were picked, dried naturally on a well-ventilated shade, and then shredded to a size of 20 to 100 mesh to obtain a dried Nipana palm flower.

One liter of water was added to 100 g of dried naphtha palm leaf powder, and the mixture was extracted at 70 ° C for 10 hours, and then filtered with filter paper to obtain an extract. The solvent was then concentrated under reduced pressure and dried, yielding 7.15 g of an extract.

(Example 2): Preparation of Extract of Hibiscus japonica and Hibiscus Extract.

The flower buds of Nipaya were picked, dried naturally on a well-ventilated shade, and then shredded to a size of 20 to 100 mesh to obtain a dried Nipana palm flower.

1 L of water was added to 100 g of the above-mentioned dried niacin palm kernel powder, and the mixture was extracted at 70 ° C for 10 hours, followed by filtering with filter paper to obtain an extract. The solvent was then concentrated under reduced pressure and dried, yielding 7.15 g of an extract.

Hibiscus safflower The flower of Parin was picked up, dried naturally on a well-ventilated shade, and crushed to a size of 20 to 100mesh to obtain a dry powder of flower of Hibiscus safflower parin.

1 L of 70% ethanol was added to 100 g of the Hibiscus dried powder, and the mixture was extracted at 40 DEG C for 20 hours, and then filtered with a filter paper to obtain an extract. The solvent was then concentrated under reduced pressure and dried to obtain 6.04 g of an extract.

Then, 5 g of the above-obtained nifel palm phloem extract and 0.5 g of the hibiscus extract were mixed.

(Test Example 1): Cell culture

Macrophage cell line RAW264.7 cells Murine KCLB (Korean Cell Line Bank) received from the pre-sale 100units / ㎖ penicillin-streptomycin and 10% fetal bovine serum (FBS) is by using a DMEM medium containing 37 ℃, 5% CO ₂ thermostat , And subculture was performed once every 3-4 days. Lipopolysaccharide (LPS, E. coli serotype 0111: B4) was purchased from Sigma and used.

(Experimental Example 2): Evaluation of cytotoxicity - MTT assay

RAW 264.7 cells were plated at a density of 1.0 × 10 ⁶ cells / ml in a 24-well plate and cultured at 37 ° C. in 5% CO ₂ , And the sample and LPS (1 / / ml) were simultaneously treated and cultured for 24 hours. After the incubation, MTT was added at a concentration of 500 μg / ml, followed by reaction at 37 ° C. for 3 hours, and then the supernatant was removed. DMSO was added to dissolve the formazan precipitate formed by reacting with live cells, and then transferred to a 96-well plate. Absorbance was measured at 540 nm.

FIG. 1 shows the results of cytotoxicity of the extract of the present invention, and FIG. 2 shows the results of cytotoxicity of the sample obtained by mixing the extract of the present invention with the extract of the present invention. As can be seen in Figures 1 and 2, all samples did not show any particular cytotoxicity.

(Experimental Example 3): Experiment to inhibit NO production

RAW 264.7 cells were plated at 1.5 × 10 ⁶ cells / mL on a 24-well plate and cultured for 18 h at 37 ° C in a 5% CO ₂ incubator. The cells were incubated for 24 h at the same time with 1 μg / ml LPS. After incubation, 100 μl of cell culture supernatant and 100 μl of Griess (1% sulfanilamide, 0.1% naphtylethylenediamine in 2.5% phosphoric acid) reagent were mixed and reacted for 10 min on a 96-well plate and absorbance was measured at 540 nm. The amount of nitrites in the sample was quantified using various standard concentrations of sodium nitrite dissolved in the same medium.

The results are shown in Fig. 3 for Example 1 and Fig. 4 for Example 2. Fig.

As can be seen from the attached drawings, it was confirmed that all of the treated samples inhibited nitrite production in a concentration-dependent manner, and that the sample of Example 2 exhibited significantly higher activity than that of Example 1 . In addition, it was judged that there was a similar inhibitory effect to that of DEX, which is known as an anti-inflammatory component.

(Experimental Example 4): Test for inhibiting the expression of IL-1β, IL-6 and TNF-α.

RAW 264.7 cells were plated at 1.5 × 10 ⁶ cells / mL on a 24-well plate and cultured for 18 hours at 37 ° C in a 5% CO ₂ incubator. The cells were co-cultured with LPS (1 μg / Respectively. After completion of the culture, the culture medium was centrifuged and IL-1β, IL-6 and TNF-α of the supernatant were quantified using an ELISA kit.

FIG. 5 shows the result of the test for inhibiting the expression of IL-1β according to Example 1, FIG. 6 shows the result of the test for inhibiting the expression of IL-1β according to Example 2, The results are shown in Fig. 7, the results of the test for suppressing IL-6 expression inhibition according to Example 2 are shown in Fig. 8, the results of the test for suppressing the expression of TNF-a according to Example 1 are shown in Fig. 9, -α expression inhibitory activity test results are shown in FIG.

As can be seen from the accompanying drawings, it was confirmed that all of the treated samples inhibited IL-1β, IL-6 and TNF-α expression in a concentration-dependent manner, and the sample of Example 2 It can be confirmed that the activity is remarkably high even at a low concentration. In addition, it was judged that there was a similar inhibitory effect to that of DEX, which is known as an anti-inflammatory component.

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 present invention is not limited to the disclosed exemplary embodiments. It will also be appreciated that many modifications and variations will be apparent to those skilled in the art without departing from the scope of the present invention.

Claims (6)

delete delete delete delete Extract of Nipah palm, extracted with water, or ethanol, or a mixture of water and ethanol as a solvent. The extract of Nipha palm is extracted with a nipah palm, a bud attached to a nipah palm, or a nipah palm Dried to a dry powder which is naturally dried in a shade, is added with a solvent in an amount of 3 to 20 times by weight of the dry weight and subjected to hot water extraction or cold extraction or warm extraction at a temperature of 20 to 100 ° C for 30 minutes to 5 days, Wherein the active ingredient is an extract obtained by cooling or extracting the extract or a solvent obtained by removing the solvent from the extract, and the lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, , Alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzo Byte, the profile further includes at least one of the hydroxy-benzoate, talc, magnesium stearate and mineral oil,
The extract of Nephila palmata is a concentrated solution concentrated under reduced pressure at 20-100 ° C. The concentrated extract or the powdered extract is used in water or ethanol or a mixture of water and ethanol,
The hibiscus extract is a hibiscus extract obtained by naturally drying a flower of a hibiscus safflower paraffin, and drying the dried flower powder. And a solvent obtained by removing the solvent from the extract at 30 to 5 days at an extraction temperature of 20 to 100 ° C,
A composition for antiinflammation using a nihon palm phalaenopsis extract and a hibiscus extract in an amount of 1: 0.1 ~ 0.2 by weight as an active ingredient.

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