KR20170126261A - Composition for inhibiting oral inflammation comprising fractions Citrus Unshiu Peel - Google Patents

Abstract

The present invention relates to a composition for inhibiting oral inflammation comprising fractions of citrus unshiu peel as an active ingredient, wherein the fractions of citrus unshiu peel is preferably a fraction obtained by fractionating a citrus unshiu peel ethanol extract with ethyl acetate, inhibits generation of nitric oxide more excellently than a citrus unshiu peel ethanol extract in periodontal ligament cells causing inflammatory, induces HO-1 expression, and inhibits inflammatory cytokine secretion. Therefore, the fractions of citrus unshiu peel of the present invention can be usefully used as toothpaste, mouthwash, a mouth spray, mouth ointment, candy or gum for inhibiting inflammation in the mouth.

Description

TECHNICAL FIELD The present invention relates to a composition for inhibiting oral inflammation comprising fragments of dermis as an active ingredient,

The present invention relates to a composition for inhibiting inflammation in the mouth such as periodontitis, gingivitis or dental caries.

Periodontal disease refers to any disease that occurs in the periodontal tissue, and can be divided into gingivitis and periodontitis depending on the severity of the disease. Among them, gingivitis is a relatively light and rapid form of periodontal disease. Gingivitis is a type of inflammation limited to the soft tissues only. Gingivitis refers to the case where the inflammation progresses to the gums and gums. This periodontal disease is a major cause of tooth loss after adulthood, and it is important to prevent and treat it in the modern age society.

Oral cleaning agents are used for the purpose of preventing oral inflammation, tooth decay and bad breath, such as periodontitis caused by damage to the oral cavity and teeth caused by changes in the eating habits of modern people. Oral detergents currently on the market are manufactured by using sodium fluoride, benzethonium chloride, polyoxyethylene, polyoxypropylene polymer, cetylpyridinium chloride, sodium saccharin, and sodium benzoate, which are antibacterial components, . However, these ingredients show side effects such as vomiting, nervous breakdown, coma, and diarrhea when swallowed, and they cause fatal damage if swallowed. Therefore, infants under the age of 7 are restricted from using mouthwash. In addition, studies have been published that cause damage to the oral cavity during long-term use.

Recently, attempts have been made to add natural products such as chitosan, aloe, green tea extract, pine leaf extract and the like to mouthwash detergent compositions, but no compositions containing dermis-derived ingredients have been studied.

1. Korean Patent Publication No. 2008-0107279.

Accordingly, it is an object of the present invention to provide a composition for inhibiting inflammation in the mouth comprising a dermis-derived component as an active ingredient.

In order to achieve the above object, the present invention is the fraction of the dermis with ethyl acetate (Ethyl acetate, EtOAc), C1-C4 alcohol, water, chloroform (chloroform, CHCl ₃₎ and any one of the solvents selected from the group consisting of a mixture thereof Wherein the composition comprises a dermal fraction obtained by the method of the present invention as an active ingredient.

According to the present invention, the fraction obtained by fractionating the ethanol extract of Citrus Unshiu Peel with ethyl acetate inhibited the production of nitric oxide (NO) more than the dermis ethanol extract in the inflammation-induced periodontal ligament cells, (heme oxygenase-1) expression and inhibited the secretion of inflammatory cytokines. Therefore, the dermis fraction of the present invention can be usefully used as toothpaste, mouthwash, mouth spray, oral ointment, candy or gum for inhibiting inflammation in the mouth.

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a flow chart for preparing the dermal fraction (CHCl ₃ , EtOAc, butanol, BuOH) or distilled water according to the present invention,
FIG. 2 is a result of thin-layer chromatography (TLC) of a dermis 70% ethanol extract, each fraction or dermis single compound,
FIG. 3 shows the results of high performance liquid chromatography (HPLC) of 70% ethanol extract of dermis,
Figure 4 shows the HPLC results of CHCl ₃ fractions of 70% ethanol extract of dermis,
Figure 5 shows the HPLC results of the EtOAc fractions of 70% ethanol extract of dermis,
Figure 6 shows the HPLC results of BuOH fractions of 70% ethanol extract of dermis,
Figure 7 shows the HPLC results of the distilled water fraction of 70% ethanol extract of dermis,
FIG. 8 shows the results of examining the effect of 70% ethanol extract of dermis on the fraction of nitric oxide (NO) or HO-1 (heme oxygenase-1)
FIG. 9 shows the results of comparing the effects of the dermis 70% EtOH extract and the dermis EtOAc fraction, wherein A is the inhibitory effect of NO production, B is the effect of inducing HO-1 expression, And D was the result of comparing the inhibitory effect of inducible NO synthase (iNOS)
FIG. 10 shows the results of confirming the effect of dermis 70% EtOH extract or dermis EtOAc fraction on inflammatory cytokine secretion,
FIG. 11 shows the results of confirming the effect of the fraction of dermis EtOAc on the expression of Nrf2 (Nuclear factor-E2-related factor) or ARE (antioxidant response element)
FIG. 12 shows the results of confirming the effect of the dermal EtOAc fraction in the secretion of inflammatory cytokines when inhibiting or inhibiting the expression of HO-1.

The invention ethyl acetate (Ethyl acetate, EtOAc), C1-C4 alcohol, water, chloroform (chloroform, CHCl _3), and effective and the resultant dermal fraction by fraction in any one solvent selected from the group consisting of a mixture thereof dermis Or a pharmaceutically acceptable salt thereof. The present invention also provides a composition for inhibiting inflammation in the mouth.

The dermis (Citrus Unshiu Peel) is a dried citrus fruit which belongs to the genus Udon and has been used as an essential herb medicine in oriental medicine.

The dermis contains 1.5 to 2% of essential oil and contains limonene, bioflavonoid, isopropenyl, toluene, 8-elemene, d-copaene, , alpha-fumarole (humulene), vitamin B and C, and the like.

Among the ingredients of the dermis, especially pharmacologically active ingredients are known as bioflavonoids. About 60 kinds of bioflavonoids have been isolated and their structure has been revealed. Over 90% of them are known as hesperidin and naringin.

It is known to have the efficacy to treat symptoms such as trimming, vomiting, nausea, indigestion, flatulence, sulphate, sea water and sputum, and diuretic action.

The dermis fraction is obtained by extracting the dermis from the dermis for 1 to 3 hours with water as a first solvent selected from the group consisting of water, C1 to C4 alcohols and a mixture thereof using a Soxhlet extraction method, It is preferably obtained by fractionation into any one second solvent selected from the group consisting of acetone, C1-C4 alcohol, water, chloroform, and mixtures thereof.

More preferably, the dermis fraction is obtained by extracting the dermis with an aqueous alcohol solution; Adding chloroform to the extracted extract, and separating into a chloroform layer and an aqueous alcohol solution layer; Adding ethyl acetate to the alcohol aqueous solution layer and separating the ethyl acetate layer.

The oral inflammation may be any one selected from the group consisting of periodontitis, gingivitis, stomatitis, nephritis and dental caries, but is not limited thereto.

The composition according to the present invention may have any one form selected from the group consisting of toothpaste, oral cleanser, oral spray, oral ointment, candy and gum, but any formulation for inhibiting inflammation in the mouth may be used Do.

In another aspect, the present invention a dermal fraction obtained by fractionation of the dermis with ethyl acetate (Ethyl acetate, EtOAc), C1-C4 alcohol, water, chloroform (chloroform, CHCl ₃₎ and any one of the solvents selected from the group consisting of a mixture thereof The present invention also provides a pharmaceutical composition for preventing or treating inflammation in the oral cavity.

The pharmaceutical compositions may further comprise suitable carriers, excipients or diluents conventionally used in the manufacture of pharmaceutical compositions.

Examples of the carrier, excipient or diluent which can be used in the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil.

The pharmaceutical composition according to the present invention may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral preparations, suppositories and sterilized injection solutions according to a conventional method .

In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose sucrose), lactose, gelatin, and the like.

In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included .

Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. 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. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The amount of the dermal fraction which is an active ingredient of the pharmaceutical composition according to the present invention may vary depending on the age, sex, body weight and disease of the patient, but it is preferably 0.001 to 100 mg / kg, preferably 0.01 to 10 mg / kg per day It may be administered once or several times.

Further, the dosage of the pharmaceutical composition according to the present invention may be increased or decreased depending on the route of administration, degree of disease, sex, weight, age, and the like. Thus, the dosage amounts are not intended to limit the scope of the invention in any manner.

The pharmaceutical composition may be administered to mammals such as rats, mice, livestock, humans, and the like in a variety of routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intratracheal, intrauterine or intracerebroventricular injections.

In another aspect, the present invention a dermal fraction obtained by fractionation of the dermis with ethyl acetate (Ethyl acetate, EtOAc), C1-C4 alcohol, water, chloroform (chloroform, CHCl ₃₎ and any one of the solvents selected from the group consisting of a mixture thereof And a health functional food for preventing or ameliorating inflammation in the mouth.

The health functional food may be provided in the form of powder, granules, tablets, capsules, syrups or beverages. The health functional food may be used in combination with other food or food additives in the active ingredient dermis fraction, Lt; / RTI > The amount of the active ingredient to be mixed can be suitably determined according to its use purpose, for example, prevention, health or therapeutic treatment.

The dermatophyte fraction contained in the health functional food may be used in accordance with the effective dose of the pharmaceutical composition. However, in the case of long-term intake for health and hygiene purposes or for health control purposes, It is clear that the component can be used in an amount of more than the above range since there is no problem in terms of safety.

There is no particular limitation on the type of the health functional food, and examples thereof include meat, sausage, bread, chocolate, snack, confectionery, pizza, ramen noodles, other noodles, dairy products including ice cream, various soups, drinks, tea, Beverages and vitamin complexes.

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, the present invention will be described in detail with reference to the following examples. However, the following examples are intended to illustrate the contents of the present invention, but the scope of the present invention is not limited to the following examples. Embodiments of the present invention are provided to more fully describe the present invention to those skilled in the art.

< Example  1> dermis extract and Fraction  purchase

1. Dermatophyte extract

The dermis (Jeju acid) of the present invention was purchased and used as a product of Daeyu Pharmaceutical Co., Ltd. (Daegu Yakseong City) After adding 1 L of 70% ethanol (EtOH) to 100 g of dried dermis, it was extracted with Soxhlet extraction method for 2 hours and repeated twice.

After extraction, the mixture was concentrated using a rotary evaporator and stored at -80 ° C for use in experiments.

2. Dermis Fraction

The dermal fraction was obtained as in Fig. First, 1 L of distilled water was added to 100 g of the dermis 70% EtOH extract prepared above to prepare a suspension, which was then added to a fraction funnel. 1 L of chloroform (CHCl ₃ ) was added thereto, followed by vortexing and shaking for 10 minutes. This was separated by allowing to stand for 24 hours, after which only the chloroform layer was obtained.

After separating the chloroform layer, 1 L of ethyl acetate (Ethyl Acetate, EtOAc) was added to the fraction funnel, followed by vortexing and shaking for 10 minutes. This was separated by allowing to stand for 24 hours, after which only ethyl acetate layer was obtained.

After separating the ethyl acetate layer, 1 L of butanol (BuOH) was added to the fraction funnel, followed by vortexing and shaking for 10 minutes. This was separated by separating for 24 hours, after which only the butanol layer was obtained.

Further, only the butanol layer was obtained and only the remaining distilled water layer was obtained.

As a result, CHCl ₃ fractions (16.7 g), EtOAc fractions (1.1 g), BuOH fractions (10.3 g) and distilled water fractions (60.5 g) were obtained in 100 g of dermis 70% ethanol extract.

< Example  2> extract or By fraction  Confirm the ingredient difference

One. Thin layer chromatography (thin-layer chromatography, TLC)

0.1 mg each of a 70% ethanol extract of dermis, each fraction or a dermis single compound [neohesperidin, acubin, quercetin] was dissolved in 1 ml of methanol (Methanol, MeOH) . The mixed solution of CHCl ₃ : MeOH: distilled water (2: 1: 0.1 volume ratio) was used as a normal phase TLC development solvent and saturated by using a mixed solution of MeOH: distilled water (9: 1 volume ratio) Respectively. After completion of the development, TLC was dried and the color reaction was confirmed with 2 ml of 10% sulfuric acid (H ₂ SO ₄ ).

As a result, as shown in Fig. 2, the Rf value of neohesperidin on TLC was found to be about 0.50 in an orange-yellow color.

In addition, it was found that the dermal 70% ethanol extract or fraction (CHCl ₃ , EtOAc, BuOH) had an orange color at an Rf value of about 0.50.

 As a result, it was found that neohesperidin was contained.

2. High Performance Liquid Chromatography (HPLC)

The dermal 70% ethanol extracts or the constituents contained in each fraction were confirmed by HPLC.

The ethanol extract of dermis 70% was diluted to a concentration of 1 mg / ml, and HPLC was measured. The results are shown in FIG.

The CHCl ₃ fraction of the 70% ethanol extract of dermis was diluted to a concentration of 1 mg / ml, and HPLC was measured. The results are shown in FIG.

The EtOAc fraction of the dermis 70% ethanol extract was diluted to a concentration of 1 mg / ml and the result was shown in FIG.

The BuOH fraction of the 70% ethanol extract of dermis was diluted to a concentration of 1 mg / ml, and HPLC was measured. The results are shown in FIG.

The distilled water fraction of the 70% ethanol extract of dermis was diluted to a concentration of 1 mg / ml, and HPLC was measured. The results are shown in FIG.

As a result, as shown in FIG. 3 to FIG. 7, it was confirmed that retention time (RT) peaks were different for each 70% ethanol extract or fraction of dermis (CHCl ₃ , EtOAc, BuOH or distilled water). Therefore, it was judged that the components contained in each fraction were different with reference to FIG. 2 to FIG.

< Example  3> Nitric oxide (NO) or HO-1 heme oxygenase -1) expression by fraction

We examined the effect of fraction on NO and HO-1 expression in inflammatory periodontal ligament cells.

For this purpose, HI-PDL cells, which are human-derived periodontal ligament cells, were purchased from the Department of Dentistry of Kyung Hee University (Teeth and Periodontal Regeneration Center). The cells were cultured in a 5% CO ₂ incubator at 37 ° C in a medium containing 10% fetal bovine serum and 1% antibiotic (Alpha-MEM), and subcultured every 2 days.

The cultured HI-PDL cells were plated at a density of 1 × 10 ⁴ cells / well in a 96-well plate, cultured, treated with 100 ng / ml of lipopolysaccharide (LPS) and cultured for 24 hours. Then 100 or 200 μg / ml of each fraction (CHCl ₃ , EtOAc, BuOH or distilled water) was treated and cultured for 24 hours. After the incubation, 100 μl of Griess reagent (1% sulfanilamide, 2.5% phosphoric acid containing 0.1% naphthylethylendiamine) was added to the supernatant (100 μl), and the mixture was incubated at room temperature for 10 minutes After the reaction, absorbance was measured at 540 nm. At this time, sodium nitrite (NaNO ₂ ) was prepared for each concentration to show a standard curve, and NO production amount was calculated.

In addition, the expression of HO-1, an important in vivo action protein for oxidative stress, was confirmed by Western blotting.

The HI-PDL cells were cultured in a 96-well plate at 3 × 10 ⁵ cells / well, treated with 100 or 200 μg / ml of each fraction (CHCl ₃ , EtOAc, BuOH or distilled water) Lt; / RTI &gt; After incubation, the supernatant was removed and washed with phosphate buffered saline (PBS). Then, 20 μl of RIPA buffer (radioimmune precipitation assay buffer) was added to dissolve the cells, followed by centrifugation (13,500 rpm, 10 minutes) to obtain proteins. Protein was quantified by Bradvord assay, electrophoresed using 8% SDS-PAGE (sodium dodecyl sulphate-polyacylamide gel electrophoresis), and transferred to a PVDF membrane (PVDF membrane) at 200 mA for 1 hour. After blocking with 5% skim milk for 1 hour, primary antibody (HO-1, Enzo (ADI-SPA0895)) was diluted 1: 1000 and allowed to react for 12 hours. After washing with Tris-Buffered Saline buffer (Tween 20) three times for 10 minutes, the cells were reacted with a secondary antibody conjugated with horseradish peroxidase, followed by ECL (enhanced chemiluminescence) And protein expression was confirmed. At this time, the amount of Actin expression was measured as a standard protein.

As a result, fraction of ethanol extract of 70% dermis (CHCl ₃ , EtOAc, BuOH or distilled water) inhibited NO production in a concentration dependent manner. In particular, the dermis EtOAc fraction was the most highly inhibitory to NO production (Fig. 8A). The highest concentration of EtOAc was then used at 200 μg / ml.

In addition, it was confirmed that the dermal EtOAc fraction in fraction (fraction of CHCl ₃ , EtOAc, BuOH, or distilled water) of dermis 70% ethanol extract induces HO-1 expression clearly (FIG.

< Example  4> dermis 70% EtOH  Extract and dermis EtOAc Fraction  compare

1. NO inhibition and induction of HO-1 expression

The effects of dermal 70% EtOH extracts and dermal ETOAc fractions were compared in NO inhibition and induction of HO-1 expression.

The HI-PDL cells treated with LPS were treated with dermis 70% EtOH extract or dermis EtOAc fraction, and the production of NO was confirmed in the same manner as in Example 3. The dermis EtOAc fraction was found to be NO (Fig. 9A).

In addition, HI-PDL cells were treated with 10, 50, 100 or 200 μg / ml of dermis 70% EtOH extract or dermis EtOAc fraction, and the expression of HO-1 was confirmed in the same manner as in Example 3 above. As a result, HO-1 expression was increased in both the dermis 70% EtOH extract and the dermis EtOAc fraction, but the dermis EtOAc fraction significantly increased HO-1 expression compared to the dermis 70% EtOH extract, and the control group, Copp, HO-1 inducer than HO-1 inducer (Fig. 9B).

2. Cytotoxicity check

The cell viability after treatment with 70% EtOH extract or dermis EtOAc fraction was confirmed by MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide).

First, HI-PDL cells were cultured in a 96-well microplate at a density of 1 × 10 ⁴ cells / well and treated with 100% or 200 μg / ml of dermis 70% EtOH extract or dermis EtOAc fraction for 24 hours. Then, 100 μl of MTT reagent (5 mg / ml concentration) was treated and reacted for 4 hours. After the reaction was completed, the supernatant was removed and 200 μl of dimethyl sulfoxide (DMSO) was treated to complete the reaction and the absorbance was measured at 590 nm. Relative cell viability was calculated assuming that the cell survival rate of the group cultured without the sample was 100%.

As a result, it was confirmed that neither the dermis 70% EtOH extract nor the dermis EtOAc fraction showed toxicity to 200 μg / ml (FIG. 9C).

3. Induction type NO production enzyme (inducible NO synthase , iNOS ) Expression inhibition confirmation

In addition, HI-PDL cells were treated with 10%, 50%, 100% or 200 μg / ml of a dermis 70% EtOH extract or dermis EtOAc fraction, and then subjected to the same Western blotting as in Example 3 to inducible NO synthase (iNOS) . At this time, an iNOS antibody (Santa Cruz (sc-650)) was reacted with a primary antibody instead of the HO-1 antibody for 12 hours.

As a result, both the dermis 70% EtOH extract or the dermis EtOAc fraction decreased the expression of iNOS in a concentration-dependent manner, but the dermis EtOAc fraction significantly decreased the expression of iNOS even at a low concentration of 10 μg / ml (FIG.

4. Inflammatory cytokine ( Cytokine ) Confirm the effect on secretion

In addition, LPS-treated HI-PDL cells were treated with dermal 70% EtOH extract or dermis EtOAc fraction to confirm the secretion of inflammatory cytokines.

First, HI-PDL cells were cultured in a 96-well microplate at a density of 1 × 10 ⁴ cells / well and cultured for 24 hours at 100 or 200 μg / ml of dermis 70% EtOH extract or dermis EtOAc fraction. Then, the supernatant was taken and the secretion amount of inflammatory cytokine tumor necrosis factor-alpha, TNF-alpha, interleukin-1 beta, IL-1 beta, IL-6 or IL- (R & D system, Minneapolis, USA) according to the manufacturer's instructions.

As a result, it was confirmed that the dermis 70% EtOH extract or the dermis EtOAc fraction suppresses the secretion amount of TNF-α, IL-1β, IL-6 or IL-12, which are inflammatory cytokines, as shown in FIG. In particular, it was confirmed that the dermal EtOAc fraction significantly inhibited the secretion of inflammatory cytokines from the 70% EtOH extract of dermis.

< Example  5> Dermis EtOAc Fraction  Check the effect

1. Identification of effects in antioxidant mechanisms

Nuclear factor-E2-related factor (Nrf2) is present in the form of a defect in other proteins, and when oxidative stress occurs in the cell, it breaks the bond and induces the action of the gene against oxidative stress.

Therefore, after treatment with EtOAc, the transcription of Cytosolic Nrf2 and Nuclear Nrf2 was confirmed.

HI-PDL cells were treated with 200 μg / ml of the dermis EtOAc fraction for 0, 0.5, 1 or 1.5 hours, followed by Western blotting in the same manner as in Example 3 to confirm the expression of Nrf2 or Nrf2 in the nucleus. At this time, Nrf2 antibody (Abcam (ab31163)) was reacted with the primary antibody for 12 hours.

As a result, it was confirmed that the expression of Nrf2 in the matrix was decreased and the expression of Nrf2 in the nucleus was increased with lapse of time (FIG. 11A). Therefore, it was confirmed that the nuclear transfer was carried out in Nrf2.

In addition, the effect of the dermis EtOAc fraction on the binding of the Nrf2-binding elements, an antioxidant response element (ARE), was confirmed.

First, the dermal ETOAc fractions were treated with 10, 50, 100 or 200 mg / ml of HI-PDL cells transformed with the ARE-luciferase gene, respectively. Transformation of HI-PDL cells using the ARE-luciferase gene was performed using methods known in the art.

As a result, the ARE binding increased as the concentration of the dermis EtOAc fraction increased, as in the expression of Nrf2 (FIG. 11B).

2. Identification of effects on NO production or inflammatory cytokine secretion

When HI-PDL cells were treated with LPS (100 μg / ml) alone, HI-PDL cells treated with LPS (100 ng / ml) were treated with only the dermis EtOAc fraction (200 μg / ml) in NO production or inflammatory cytokine secretion In one case, the treatment of HI-PDL cells treated with LPS (100 ng / ml) was compared with the simultaneous treatment of the dermis EtOAc fraction and the HO-1 inhibitor SnPP (50 μM).

NO production was confirmed in the same manner as in Example 3, and inflammatory cytokine secretion was confirmed in the same manner as in Example 4-4.

As a result, as shown in Fig. 12, it was confirmed that when the dermis EtOAc fraction was treated with inflammation-induced cells, the NO production or the expression of inflammatory cytokines such as TNF- ?, IL-1 ?, IL-6 and IL- Respectively. On the other hand, when the expression of HO-1 was inhibited, the expression of NO production, TNF-α, IL-1β, IL-6 and IL-12 was increased again. Therefore, the anti-inflammatory effect of the dermal EtOAc fraction was found to be HO-1 mediated.

While the present invention has been particularly shown and described with reference to specific embodiments thereof, those skilled in the art will appreciate that such specific embodiments are merely preferred embodiments and that the scope of the present invention is not limited thereby. something to do. It is therefore intended that the scope of the invention be defined by the claims appended hereto and their equivalents.

Claims (7)

Including a dermal fraction obtained by fractionation of the dermis with ethyl acetate (Ethyl acetate, EtOAc), C1-C4 alcohol, water, chloroform (chloroform, CHCl ₃₎ and any one of the solvents selected from the group consisting of a mixture thereof as an active ingredient Or a pharmaceutically acceptable salt thereof. The method according to claim 1,
The dermis fraction is obtained by extracting the dermis from the dermis for 1 to 3 hours with water as a first solvent selected from the group consisting of water, C1 to C4 alcohols and a mixture thereof using a Soxhlet extraction method, Wherein the composition is obtained by fractionation into any one second solvent selected from the group consisting of acetone, C1-C4 alcohol, water, chloroform, and mixtures thereof. The method according to claim 1,
The dermis fraction
Extracting the dermis with an aqueous solution of alcohol;
Adding chloroform to the extracted extract, and separating into a chloroform layer and an aqueous alcohol solution layer;
Wherein the ethyl acetate fraction is an ethylacetate fraction obtained by adding ethyl acetate to the alcohol aqueous solution layer and separating the ethyl acetate layer. The method according to claim 1,
Wherein the oral inflammation is any one selected from the group consisting of periodontitis, gingivitis, stomatitis, nephritis, and dental caries. The method according to claim 1,
Wherein the composition has any one of the formulations selected from the group consisting of oral cleanser, toothpaste, mouth spray, oral ointment, candy and gum. Including a dermal fraction obtained by fractionation of the dermis with ethyl acetate (Ethyl acetate, EtOAc), C1-C4 alcohol, water, chloroform (chloroform, CHCl ₃₎ and any one of the solvents selected from the group consisting of a mixture thereof as an active ingredient Or a pharmaceutically acceptable salt thereof. Including a dermal fraction obtained by fractionation of the dermis with ethyl acetate (Ethyl acetate, EtOAc), C1-C4 alcohol, water, chloroform (chloroform, CHCl ₃₎ and any one of the solvents selected from the group consisting of a mixture thereof as an active ingredient Or a pharmaceutically acceptable salt thereof.

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