KR20130060084A - Composition for preventing or treating peridontal disease - Google Patents

Abstract

PURPOSE: A pharmaceutical composition containing a Cudrania tricuspidata extract is provided to prevent or treat periodontal diseases. CONSTITUTION: A pharmaceutical composition for preventing or treating periodontal diseases contains 0.01-0.05wt% of a Cudrania tricuspidata extract as an active ingredient. The extract is prepared from Cudrania tricuspidata leaves using water, methanol, ethanol, butanol, ethyl acetate, or a mixture thereof. The extraction is performed by room temperature extraction, hot water extraction, maceration, reflux and cooling extraction, sonication, or vapor extraction. [Reference numerals] (AA) Cudrania tricuspidata 0.5%; (BB) Cudrania tricuspidata 1%; (CC) Cudrania tricuspidata 2%; (DD) Cudrania tricuspidata 2.5%

Description

Composition for preventing or treating peridontal disease

The present invention relates to a pharmaceutical composition, toothpaste composition, oral cleansing composition or oral protection agent for the treatment or prevention of periodontal disease comprising kkujippong extract as an active ingredient.

Periodontal disease, also known as taste, is divided into gingivitis and periodontitis, depending on the severity of the disease. It is a relatively light and fast-recovering form of periodontal disease that is limited to the gums, ie soft tissues, is called gingivitis, and when this inflammation progresses to the gums and around the gum bone, it is called periodontitis.

There is a V-shaped gap between the gingiva (gingiva) and the teeth, which is a periodontal disease that bacteria attack the area below the gum line of the sulcus to damage the periodontal ligament and adjacent tissues. As the inflammation progresses and more tissue is damaged, the groove develops into a periodontal pocket, and the more severe the periodontitis, the deeper the periodontal sac becomes. As the periodontal sac becomes deeper, the periodontal ligament becomes inflamed and bone loss occurs.

Periodontal disease is closely related to age, with more than half of adults over 20 years old, three out of four after 35 years of age, and 80-90% of elderly people over 40 years old. A direct cause of periodontal disease is a bacterial film called plaque that is formed continuously in the teeth. Plaques are sticky and colorless, and if they are hardened and not removed, they become tartar.

When plaque and tartar accumulate, the gum falls from the tooth, which causes a gap to open, forming a periodontal pocket between the tooth and the gum. As inflammation progresses, the gap between the gums and teeth increases, and the alveolar bone and periodontal ligaments are destroyed. Malnutrition of proteins and vitamins, hormonal disorders such as pregnancy or diabetes, smoking and AIDS can make the disease worse.

Currently, antimicrobial agents are used to treat such oral diseases. In general, dental caries uses chlorhexidine 0.2% nutrient solution for plaque management, but it is used only for short-term therapy due to side effects and unpleasant taste of tooth stains. In periodontal disease, calculus removal is primarily performed, but as an adjuvant therapy, more microorganisms are involved in treating antimicrobial agents than caries, so tetracycline and metronidazole are used in addition to chlorhexidine toothpaste. . However, these antimicrobial agents can cause side effects, such as the generation of resistant bacteria, non-selective sterilization, it is necessary to find an effective substance that can replace the existing antimicrobial agents.

Although antibiotics such as penicillin are used to inhibit the growth of bacteria causing the formation of dental plaques for the purpose of preventing dental caries, they are not used in clinical practice due to their resistance to prolonged use (Kwak, Dong-Ju, Korean Journal of Hygiene and Science, pp. 99-). 107, 2004). In addition, methods of using toothbrushes and other fluorine compounds, fluorine-releasing devices, and automatic toothbrush instruments have been developed and introduced. Effect and isolation of antimicrobial substance brazilin Wonkwang Dental Clinic, 2004) To date, it is evidence that dental caries is a major cause of major tooth loss. For this reason, the development of a more effective and stable dental caries prevention method is required.

In Korea, natural products are attracting attention as alternatives with less side effects and improved therapeutic effects. Sanguinaria extract (US Pat. No. 4,689,216), Ear woowoo extract (Korean Patent No. 10-2002-0082308), Schisandra extract (Korean Patent No. 10-2000-0047165), and the like. In the case of a therapeutic agent using natural product extracts and components that have been used and proven to be harmless to humans since ancient times, resistance or safety problems caused by existing therapeutic agents can be overcome. In particular, herbal functional items using natural products that have been used as herbal medicine are popular. In the case of tooth decay, brushing and oral cleansing are important, so developing toothpaste or mouthwash containing natural ingredients with antibacterial properties to tooth decay can contribute to health and economic vitalization.

In addition, because gingivitis and periodontitis are accompanied by an inflammatory response, consideration should also be given to the preparation of a more effective composition for inhibiting inflammation.

Accordingly, the present inventors, while studying natural medicines for treating or preventing the periodontal disease as described above, the present invention was completed by discovering that the kkujippong extract inhibits the growth of periodontal causative bacteria and inhibits the inflammatory response.

The present invention relates to a pharmaceutical composition, toothpaste composition, oral cleaning agent for the treatment or prevention of periodontal disease comprising kkujippong extract as an active ingredient.

In order to solve the above problems, the present invention provides a pharmaceutical composition for the treatment or prevention of periodontal disease comprising kkujippong extract as an active ingredient.

As used herein, the term "Kujippong" refers to a kkujippong tree, and the scientific name is Cudrania tricuspidata. It is also called kkujippong. Cudrania tree is a deciduous tree belonging to the Mulberry family and is also called a mulberry tree. In the present invention, the region of the Cudrania tree used to obtain the extract is preferably the leaf portion (herb) of the Cudrania tree.

As used herein, the term "extract" refers to a specific component that has been separated from the Cudrania tree using a solvent of the liquid. As the solvent, water, methanol, ethanol, or a mixture thereof may be used. In order to efficiently extract the active ingredient, it is preferable to extract with methanol, ethanol or a mixture of these and water. The concentration of methanol and ethanol is preferably 10 to 90%, but is not limited thereto. It is even more preferable to extract using water as the solvent.

The extraction method of the present invention may use any one selected from the group consisting of hot water extraction, cold extraction, reflux cooling extraction, ultrasonic extraction and steam extraction.

The term "periodontal disease" used in the present invention, also referred to as flavor, is divided into gingivitis and periodontitis (periodontitis) depending on the degree of the disease. It is a relatively light and fast-recovering form of periodontal disease that is limited to the gums, ie soft tissues, is called gingivitis, and when this inflammation progresses to the gums and around the gum bone, it is called periodontitis.

Growth inhibition of Streptococcus mutans, one of the causative agents of periodontal disease, can be measured to determine whether periodontal disease is treated or prevented.

In addition, the present invention provides a toothpaste (toothpaste), oral cleansing agent (Gagle) comprising the kkujippong extract as an active ingredient using the treatment and prevention efficacy of periodontal disease of kkujippong extract.

In the present invention, it was confirmed that kkujippong extract is effective in the treatment or prevention of periodontal disease. According to one embodiment of the present invention, by confirming that the growth of Streptococcus mutans, which is an indicator of the treatment of periodontal disease, is inhibited, it was confirmed that there is an effect of preventing or treating periodontal disease.

The term "prevention" as used in the present invention means all actions of inhibiting or delaying the disease by administration of the kkujippong extract. In addition, the term "treatment" used in the present invention means all actions that improve or cure the symptoms of the disease by administration of the composition containing the kkujippong extract.

In addition, the present invention provides a composition for the treatment or prevention of periodontal disease, characterized in that it comprises 0.01 to 0.05% by weight of the kkujippong extract based on the total weight of the composition.

The composition for the treatment or prevention of periodontal disease according to the present invention inhibits the production of acid by oral microorganisms, and effectively inhibits the growth of oral microorganisms to inhibit dental caries and periodontal disease.

Toothpaste composition of the present invention, oral cleaning agent (Gagle) as a medicinal ingredient as a medicinal ingredient sodium fluoride, sodium fluoride phosphate, amine fluoride, tin fluoride or xylitol which is a caries prevention agent; Fungicides chlorohexidine, cetylpyridium chloride or triclosan; Herbal medicines such as bamboo salt or propolis; Anti-inflammatory agents allantoin, aminocaproic acid, tranexamic acid, tocopherol acetate or methylsulfonylmethane; and enzymes dextranase, glucose oxidase, glucoamylase, α, β-amylase, lactoperoxidase and lysozyme It may further comprise any one or more components selected from the group.

In addition, the toothpaste composition of the present invention, oral cleansing agent may further include any one or more materials selected from the group consisting of abrasives, wetting agents, binders, foaming agents, sweetening agents, anionic surfactants, fragrances and pigments. The composition may be in the form of a paste, powder, liquid (gagle) or powder.

In addition, the toothpaste composition of the present invention, the mouthwash is an abrasive, precipitated silica, silica gel, zirconium silicate, calcium dihydrogen phosphate, anhydrous calcium monohydrogen phosphate, hydrous alumina, hard calcium carbonate, heavy calcium carbonate, calcium pyrophosphate, insoluble methphosphate And it may further comprise any one or more abrasives selected from the group consisting of aluminum silicates. Preferably, the abrasive comprises 5 to 50 weight percent based on the total weight of the composition.

In addition, the degree of polishing of the final product exhibits a value of 30 to 100 so that there is no tooth wear due to excessive polishing, it is preferable to use a particle size of 20 ㎛ or less, although the particle size also varies depending on the type of abrasive. In general, the polishing value starts at 0 and represents a relative polishing degree with a standard toothpaste of 100. In general, most products have a value of 200 or less.

In addition, the toothpaste composition of the present invention, oral cleansing agent is any one selected from the group consisting of concentrated glycerin with a concentration of 98%, aqueous solution of sorbitol at a concentration of 70%, aqueous solution of amorphous sorbitol at a concentration of 70%, xylitol, polyethylene glycol and propylene glycol. It may further comprise one or more wetting agents. The humectant preferably comprises 20 to 70 wt% based on the total weight of the composition.

The humectant is an essential base ingredient for making ointment formulations, which prevents the toothpaste from drying and solidifying when exposed to air and provides shine to the surface of the toothpaste. .

In addition, any kind of water-soluble polymer may be used as the binder included in the toothpaste composition and oral cleaning agent of the present invention. Preferably, sodium carboxymethyl cellulose synthesized from cellulose of wood, carrageenan or microorganism extracted from algae Xanthan gum obtained from metabolism can be used. The binder serves to prevent separation of the solid powder and liquid components.

In addition, as the foaming agent included in the toothpaste composition and oral cleaning agent of the present invention, it is preferable to use sodium lauryl sulfate, an anionic surfactant, and a copolymer of polyoxyethylene polyoxypropylene (poloxamer) depending on the characteristics of the formulation. ), Nonionic surfactants such as polyoxyethylene hardened castor oil or polyoxyethylene sorbitan fatty acid ester can be used. The foaming agent enhances the feeling of use of the product and helps cleansing action, quickly disperses and penetrates the other active ingredients, and reduces the interfacial tension to act to easily remove foreign substances in the oral cavity.

In addition, the toothpaste composition of the present invention, flavors, sweeteners, pigments and the like contained in the oral cleanser serves to improve the feel of the toothpaste composition.

Perfume included in the toothpaste composition of the present invention, oral cleansing agent is preferably edible, and examples thereof include menthol, peppermint oil, spearmint oil, sage, eucalyptol, methyl salicylate or fruit extract. In addition, it is preferable to use sodium saccharin as the sweetener, and food coloring is preferably used as the dye.

Composition for the treatment or prevention of periodontal disease comprising kkujippong extract of the present invention as an active ingredient according to one embodiment of the present invention, the survival rate of the cells was maintained in the evaluation of cytotoxicity as an indicator of stability, the stability Confirmed.

In addition, the composition for the treatment or prevention of periodontal disease comprising the kkujippong extract of the present invention as an active ingredient, according to an embodiment of the present invention, the organic acid production of Streptococcus mutans which is one indicator for confirming dental caries Having inhibitory activity, it was confirmed that there is a prevention or treatment effect of dental caries.

The present invention relates to a pharmaceutical composition, toothpaste composition or oral cleaning agent for the treatment or prevention of periodontal disease comprising kkujippong extract as an active ingredient, it can prevent or treat periodontal disease with the composition according to the present invention.

Figure 1 shows the antimicrobial activity of Streptococcus mutans (S. mutans) of the kkujippong extract according to an embodiment of the present invention. (A) is a control group treated with sterile distilled water, (B) shows the growth inhibition ring when treated with Angelica extract and (C) shows the growth inhibition ring when treated with kkujippong extract.
Figure 2, showing the antimicrobial activity of the concentration of kkujippong extract for the growth of time Streptococcus mutans (S. mutans) according to an embodiment of the present invention.
Figure 3 shows the pH change over time by the addition of the extract of Streptococcus mutans (S. mutans) kkujippong according to an embodiment of the present invention.
Figure 4 shows the cell viability (%) by the addition of kkujippong extract according to Example 4 of the present invention.
Figure 5 shows the inhibition of NO production (uM) by the addition of the kkujippong extract according to Example 4 of the present invention.
Figure 6 shows the inhibition of NO production (uM) by the addition of Aspirin, a control for comparison with the effect of kkujippong extract of FIG.

Hereinafter, the present invention will be described in more detail with reference to the following Production Examples and Examples. However, the following Preparation Examples and Examples are for illustrating the present invention, but the scope of the present invention is not limited thereto.

Example  : CJ  Preparation of extract

After lyophilizing 10 g of kkujippong extract stock solution, the mixture was extracted by heating at reflux at 85 ° C. for 3 hours, cooled, and filtered through a filter paper. After freeze-drying the filtered extract was obtained 0.011g of kkujippong extract.

Reference Example  One

Streptococcus mutans (KCTC 3065, Korea Life Resource Center), a strain used in the present invention, was cultured in BHI medium (Brain heart infusion, Difco, USA) and mixed well with glycerol (glycerol) in a ratio of 3: 7- It was used for the experiment while storing in a 70 ℃ freezer. Strains stored in the freezer were activated three days before the start of the experiment and inoculated in a BHI agar plate or BHI liquid medium and used for experiments incubated in a thermostat at 37 ℃ for 24 hours.

Experimental Example  1: the causative agent of tooth decay Streptococcus Mutans  Growth inhibitory effect measurement

1-1) Hole  Diffusion method hole diffusion method )On by Of clear zone  Measure

In order to measure the clear zone by the kkujippong extract against Streptococcus mutans, the causative agent of tooth decay, the experiment was performed as follows using a hole diffusion method according to the standard method of the National Committee for Clinical Laboratory Standards (NCCLS). It was. Streptococcus mutans strain was pre-cultured for 24 hours by the method of Reference Example 1, prepared at a concentration of 1 × 10 ⁸ CFU / ml, and 1 mL in 5 mL of top agar (0.7% agar) maintained at 45 ° C. After adding well, the mixture was layered on a BHI agar plate (1.5% agar) that was hardened in advance. Sterile stainless steel hole maker (diameter 8mm) was raised before the top was hardened to make holes. The extract was applied to 70 ul of the stock solution. As a control, the same amount of sterile distilled water used as the solvent of the extract was used. In addition, the same experiment was performed by applying 70ul (final 490 ug) per hole of the sample of the Angelica extract 7mg / ml concentration for comparison with the Angelica extract known to have a streptococcus mutans growth inhibitory effect. The growth inhibition produced after 48 hours of incubation at 37 ° C. was measured using micrometa (Mitutoyo, Kawasaki, Japan).

As a result, as shown in Figure 1 (A) sterile distilled water treated control group did not show the growth inhibitory effect on the decay bacteria, (B) in the 490 ug treated group of Angelica extract extract growth inhibition of 12.0 ± 0.5 mm produced (C) In the case of the Cudrania extract treatment group, growth inhibition ring of 16.0 ± 0.5 mm was generated. Therefore, kkujippong extract has an antimicrobial effect against Streptococcus mutans, the causative agent of tooth decay, which was confirmed to be more effective than Angelica extract (FIG. 1, Table 1).

Hole diffusion method Extract (A) H ₂ O (B) Angelica (C) Cudrania extract Volume 70 ul 70 ul of 7 mg / ml
(490 ug) 70 ul Inhibitory zone (mm) 8 ± 0 12 ± 0.5 16 ± 0.5

1-2) Determination of antibacterial activity against growth inhibition by liquid culture method

Direct growth inhibition activity in liquid culture of Cudrania extract against the causative bacteria Streptococcus mutans was measured according to the standard method of the National Committee for Clinical Laboratory Standards (NCCLS). Streptococcus mutans bacteria prepared by culturing for 1 hour at 1 × 10 ⁶ CFU / ml were pre-cultured in a medium containing 1% of glucose-sterilized glucose in BHI medium to a final concentration of 5 × 10 ³ CFU. / inoculated with / ml zigzag extract was added to the final concentration of 0, 0.5, 1.0, 2.0, 2.5%, respectively, and cultivated for 27 hours at 37 ℃ incubator and spectrophotometry at 600 nm of the bacterial culture at 3 hours intervals It was measured using a photometer. As a result, as shown in FIG. 2, in the control group without addition of the zigzag extract, the growth of the mutans bacteria reached a maximum of 1.86 after 12 hours of incubation, but the final absorbance after 27 hours in the experimental group to which 0.5% of the zigzag extract was added. Was 1.64, and 1%, 2%, and 2.5% of the zigzag extract addition group showed 1.21, 1.09, and 0.99, respectively, indicating strong antimicrobial activity (FIG. 2).

Experimental Example  2: Streptococcus Mutans  Review of organic acid production inhibitory activity

As shown in Experiment 1, 0.22 um filtered sterilized glucose was added to the BHI liquid medium to a final concentration of 1%, and then, each sample was added by concentration, and inoculated with Streptococcus mutans bacteria for 27 hours at 37 ° C. in a thermostat. While culturing, a predetermined amount of the culture solution was collected at 3 hour intervals and the pH of the supernatant after centrifugation was measured using a pH meter (EUTECH PH-6, EUTECH, SINGAPORE) to observe the effect of inhibiting organic acid production. As a result, as shown in Fig. 3, the culture medium of Streptococcus mutans was rapidly lowered at the initial culture of pH 7.3 at 12 hours of culturing, and recorded pH 4.4 at 27 hours of culturing. On the other hand, the zigzag extract started slowly showing the suppression of the organic acid production of the mutans from 12 hours of culture, and showed the inhibition of the organic acid production in a concentration-dependent manner.

Experimental Example  3: RAW  264.7 Cytotoxicity in Cells and NO  Generation suppression activity test

Cujippong extract was obtained in the same manner as in the above example.

3-1) Cell Culture

RAW 264.7 cells, mouse macrophage lines, were treated at 37 ° C. and 5% CO ₂ in DMEM (Dulbecco's Modified Eagle Medium) medium containing 10% fetal bovin serum (FBS) and 1% streptomycin-penicillin. The culture was used in the following experiment.

3-2) Reagents Used

3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl-tetrazolium bromide [3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl-tetrazolium bromide , MTT], lipopolysaccharides (LPS, Escherichia coli), grease 1% sulfanylamide (in 5% phosphoric acid) and 0.1% naphthylenediamine dihydrochloride were used in the experiment.

3-3) Cytotoxicity test method

On the first day, the cultured RAW 264.7 cells were incubated for 24 hours in 1.0 × 10 ⁵ cells / well in 24 well plates. On the second day, the medium was removed and the Kuzzangpon extracts were treated at concentrations of 0, 10, 50, 75 and 100 ug / ml, respectively. This is 37 ℃, 5% CO ₂ After the reaction in the incubator for 24 hours, the culture solution was removed. On the third day, the culture solution was removed and MTT agent (5 mg / ml) was added at a concentration of 0.5 mg / ml and reacted for 3 hours in a 37 ° C., 5% CO ₂ incubator. In-plate culture was removed. 2 ml of dimethyl sulfoxide (DMSO) was added to dissolve the precipitate, and the absorbance was measured at 540 nm.

3-4) Grease ( Griess ) Reaction NO  Production Inhibition Activity Test Method

On the first day, the cultured RAW 264.7 cells were incubated for 24 hours in 1.0 × 10 ⁵ cells / well in 24 well plates. On the second day, the medium was removed and treated with 1 ug / ml of LPS (Escherichia coli 026: B6; Difco, Detroit, MI, USA), and then the zigzag extracts were prepared at 0, 10, 50, 75, and 100 ug / ml concentrations, respectively. Treated. This was reacted for 24 hours at 37 ℃, 5% CO ₂ incubator and the culture was removed. On the third day, intraplate medium was dispensed in 100 ul into 96 well plates. NO production was measured by the amount of nitrite released from the cells. After reacting the same amount of grease (Griess) in the medium, the absorbance was measured at 540 nm.

3-5) RAW  264. Cytotoxicity test results of samples in 7 cells

As shown in FIG. 4 and Table 2, the cytotoxicity of each sample in the RAW 264.7 cells was analyzed by MTT assay.

(ug / mL) CJ (SD) LPS (-) 0 81.22 1.27 LPS (+) 0 100.00 0.15 10 100.06 0.58 50 97.74 2.05 75 89.25 1.89 (unit %) 100 88.28 1.30

3-6) RAW  264. Of Samples in 7 Cells NO  Production Inhibitory Activity Test Results

As shown in FIG. 5 and Table 3, after treating lipopolysaccharide (LPS), which is a representative inflammation-inducing substance, to RAW 264.7 cells at a concentration of 1 ug / ml, NO production inhibitory activity by each sample was analyzed through a grease reaction. As a result, the anti-inflammatory effect was confirmed by suppressing NO production in concentration-dependent manner compared to the LPS-treated group in the Cudrania extract.

(ug / mL) CJ (SD) LPS (-) 0 1.40 0.30 LPS (+) 0 33.47 1.49 10 28.61 0.27 50 22.85 0.25 75 15.96 0.17 (Unit uM) 100 15.34 0.15

Claims (8)

Pharmaceutical composition for the treatment or prevention of periodontal disease comprising kkujippong extract as an active ingredient.
According to claim 1, wherein the kkujippong extract is a composition, characterized in that extracted from the leaves of kkujippong.
The composition of claim 1, wherein the zigzag extract is extracted with water, methanol, ethanol, butanol, ethyl acetate, or a mixture thereof.
The composition according to claim 1, wherein the extraction is room temperature extraction, hot water extraction, cold needle extraction, reflux extraction, ultrasonic extraction or steam extraction.
According to claim 1, wherein the composition is characterized in that it comprises 0.01 to 0.05% by weight of the kkujippong extract based on the total weight of the composition.
The composition of claim 1, wherein the composition is a toothpaste or mouthwash composition.
7. Sodium fluoride, sodium fluoride phosphate, amine fluoride, tin fluoride, xylitol, chlorohexidine, cetylpyridium chloride, triclosan, bamboo salt, propolis, allantoin, aminocaproic acid, tranexamic acid Tocopherol acetate, methylsulfonylmethane, dextranase, glucose oxidase, glucoamylase, α, β-amylase, lactoperoxidase and lysozyme further comprises a composition further comprising any one or more components.
The composition of claim 1, wherein the periodontal disease is gingivitis or periodontitis.

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