KR20140041251A - Pharmaceutical composition for preventing or treating peridontal disease comprising extract of lignum acronychiae containing nerolidol - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating a periodontal disease containing Lignum Acronychiae extract as an active ingredient, a pharmaceutical composition for preventing or treating a periodontal disease containing nerolidol or pharmaceutically acceptable salts thereof as an active ingredient, a composition for washing an oral cavity containing the Lignum Acronychiae extract as an active ingredient, a composition for washing oral cavity containing the nerolidol or the pharmaceutically acceptable salts thereof as an active ingredient, a food composition for preventing or improving the periodontal disease containing the Lignum Acronychiae extract, and a food composition for preventing or improving the periodontal disease containing the nerolidol or the pharmaceutically acceptable salts thereof. The compositions containing the Lignum Acronychiae extract and the nerolidol extracted from the Lignum Acronychiae extract or the pharmaceutically acceptable salts thereof in the present invention have no side effects thereby capable of safe employment. Moreover, the compositions have an excellent and specific antibacterial activity against periodontal bacteria, thereby capable of being widely used for more effective prevention or treatment of the periodontal disease. [Reference numerals] (AA) Viable cell count (%)

Description

Pharmaceutical composition for preventing or treating peridontal disease comprising extract of Lignum Acronychiae containing nerolidol}

The present invention relates to a pharmaceutical composition for preventing or treating periodontal disease, including the Gangjin-hyang extract comprising Neroolidol, and more specifically, the present invention provides a pharmaceutical for the prevention or treatment of periodontal disease, including Kangjin-hyang extract as an active ingredient A pharmaceutical composition for the prevention or treatment of periodontal disease, comprising a composition, nerolidol or a pharmaceutically acceptable salt thereof as an active ingredient, a composition for oral cleansing, including gangjinhyang extract as an active ingredient, nerolidol or Oral cleaning composition comprising a pharmaceutically acceptable salt thereof as an active ingredient, periodontal disease comprising a food composition for the prevention or improvement of periodontal disease, including gangjinhyang extract and nerolidol or a pharmaceutically acceptable salt thereof It relates to a food composition for the prevention or improvement of.

Periodontal disease refers to a disease that damages the periodontal tissue. Typically, the periodontal tissue is composed of alveolar bone, gingiva and periodontal ligament, the periodontal disease is usually caused by the periodontal infection of the gingiva, the lesions are expanded, the periodontal ligament and alveolar bone is damaged, the symptoms worsen, bleeding hemorrhage, Various pathological symptoms such as swelling, periodontal sac formation, loss of adhered gingiva, destruction of alveolar bone, and bad breath result in loss of teeth. The periodontal disease is divided into gingivitis and periodontitis according to the area of the lesion, and gingivitis refers to a disease in which the lesion occurs only in the gum, that is, soft tissue, and periodontitis refers to the lesion. Refers to a condition that manifests itself in areas that extend to the gums and around the gum bone.

Periodontal disease is known to be highly related to systemic diseases such as atherosclerosis, myocardial infarction, stroke, diabetes, premature birth, low weight, abortion and preeclampsia (Desvarieux, M. et al., Circulation, 2005, 111, 576-582; Offenbacher , S. et al., J. Periodontol., 1996, 67, 1103-1113; Garcia, RI et al., Periodontol. 2000, 2001, 25, 21-36; Champagne, CM et al., J. Int. Acad, Periodontol., 2000, 2, 9-13; Paquette, DW, J. Int. Acad. Periodontol., 2002, 4, 101-119), various studies are being actively conducted to prevent or treat it.

As a method of treating periodontal disease, a surgical method is generally used. In the case of gingivitis, it is to be treated by using physical treatment (tartar removal and root root removal) to remove plaque and tartar of a part affected by periodontal disease. In the case of periodontitis, it may be treated by surgical operation (gingiva peeling), but in severe cases, the tooth may need to be removed. In contrast, as a method of preventing periodontal disease, surgical methods such as scaling to remove tartar on the upper part of the junctional epithelium from the surface of the teeth and non-surgical methods of treating antibacterial agents against periodontal bacteria are used. do. However, when the antimicrobial agent is used, there is a disadvantage in that side effects occur at other parts of the body when used for a long time, and studies to develop alternatives having improved or improved pharmacological effects without reducing or showing side effects are continued.

As part of this research, natural products are attracting attention. Sanguinaria extract (U.S. Patent No. 4,689,216), ear worm extract (Korean Patent Publication No. 10-2002-0082308), Schisandra extract (Korean Patent Publication No. 10-2000-0047165), etc. It has been reported to have antimicrobial activity against periodontal bacteria, although it has been proved to be harmless to humans. However, compared with the conventional antimicrobial agents, the natural products have a disadvantage in that they exhibit relatively low antimicrobial activity and exhibit antimicrobial activity against not only periodontal bacteria but also other beneficial bacteria.

Under this background, the present inventors, in order to develop a material derived from natural products that exhibits excellent antibacterial activity against periodontal bacteria but also exhibits periodontal bacteria specific antimicrobial activity, as a result of intensive research, it is isolated from the extract of Gangjinhyang and Kangjinhyang extract. It was confirmed that the nerolidol showed excellent and specific antibacterial activity against the causative agent of periodontal disease, and completed the present invention.

One object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of periodontal disease, including the Kangjin-hyang extract as an active ingredient.

Another object of the present invention is to provide a pharmaceutical composition for preventing or treating periodontal disease, including nerolidol or a pharmaceutically acceptable salt thereof as an active ingredient.

Still another object of the present invention is to provide a food composition for preventing or improving periodontal disease, including the extract of Gangjin flavor.

Still another object of the present invention is to provide a food composition for preventing or ameliorating periodontal disease, including nerolidol or a pharmaceutically acceptable salt thereof.

Still another object of the present invention is to provide a composition for oral cleansing, including the Gangjin flavor extract as an active ingredient.

Still another object of the present invention is to provide a composition for oral cleaning comprising nerolidol or a pharmaceutically acceptable salt thereof as an active ingredient.

As an embodiment for achieving the above object of the present invention, the present invention is a prevention of periodontal disease comprising a strong jinyang extract, nerolidol or pharmaceutically acceptable salts thereof isolated from the kangjinhyang extract as an active ingredient Or it provides a pharmaceutical composition for treatment.

The term "earthquake direction (降眞香of the present invention; Lignum Acronychiae ) is a deciduous tree belonging to the legume (Leguminosae), Dalbergia. odorifera T. Chen A core material with the bark of the roots removed, sometimes called Gangjin or Kanghyang. The main ingredients are dalbergin, nordalbergin, isodlbergin, dlbegenone, dalbergichromene, acronylin, acrovestone ), Dimethylacrovestone 1, bergapten, dictamnine, skimmianine, evolitrine, kokusaginine, baurenol And hydroxyobtustyrene, isomucronustyrene, methyl-2-hydroxy-3,4-dimethoxybenzoic acid, and the like. dimethoxybenzoate), (+)-duartin, (-)-melilotocarpan C, (-)-melilotocarpan D4, (-)-methylnisolin, (-)-Odoricarpan, 3'-methoxydaidzein, 3 (R) -5'-methoxyvestitol, (3R-type) -claussequinone ), Dalbergia flavan dimer Do-18, dalbergia flavan dimer Do-19, dalbergia flavan dimer Do-20, dalbergia flavan dimer ( dalbergia flavan dimer Do-21, dalbergia-bis-isoflavan, formononetin, (dl-type) -isoduartin, isoliquiritigenin, Isomedicarpin, (dl-type) -medicapin, (dl-type) -mucronulatol, muturonulatol, obtustyrene, odoriflavene, (3R-type ) -Vestitol, (dl-type) -vestitol, nerolidol, 2-aryl-2 ', 6-dihydroxy-4'-meth Methoxybenzofuran, 6-dimethylbignafuran, 6-hydroxy-2- (2-hydroxy-4-methoxyphenyl) benzofuran , β-bisabolene, β-farnesen e), baurenol and the like.

The term "Gangjinhyang extract" of the present invention means an extract obtained by extracting the Kangjinhyang with water or an organic solvent, and may be an active ingredient of the composition for the prevention, treatment or improvement of periodontal disease for the purposes of the present invention. . The Kangjinhyang extract is preferably water extracted using a solvent such as alcohol having 1 to 6 carbon atoms, nucleic acid, chloroform, methyl acetate, butanol. More preferably, the Kangjinhyang extract of the present invention may be extracted using water, an alcohol having 1 to 6 carbon atoms or a mixed solvent thereof, and most preferably Kangjinhyang hot water extract or 70% ethanol extract. It may be, but is not particularly limited thereto.

As used herein, the term "nerolidol" refers to a kind of sesquiterpene-type compound found in essential oils of various kinds of plants and flowers as an ingredient contained in extracts of Gangjin-hyang. Chemically represented as 3,7,11-trimethyl-1,6,10-dodecatrien-3-ol (3,7,11-Trimethyl-1,6,10-dodecatrien-3-ol) It is also called peruviol. The nerolidol is not particularly limited thereto, but may be preferably separated from the gangjinhyang extract. In one embodiment of the present invention was confirmed the antimicrobial activity of periodontal bacteria of nerolidol isolated from Kangjin flavor extract.

The term "pharmaceutically acceptable salts" of the present invention means acid addition salts or base addition salts suitable for the treatment of patients. For the purpose of the present invention, the salt may be used for the purpose of preventing periodontal disease or treating an individual suffering from periodontal disease in an individual that is likely to develop periodontal disease by adding an acid or a base to nerolidol, It is not particularly limited to this.

As used herein, the term "periodontal disease" refers to a term that refers to an alveolar bone or a disease affecting the alveolar bone surrounding tissue that supports and binds the alveolar bone to the gingiva, and more specifically, the periodontal infected with the alveolar bone or surrounding tissue The pathogenic metabolites secreted by the bacteria cause lesions that damage the structure of the tooth and the gingival tissue and weaken the immunity of the gingival tissue, and promote the growth of periodontal bacteria in the lesion area, By this means a disease caused by repeating the process of expanding the lesion site where the virulence metabolites are secreted. The periodontal disease is divided into gingivitis and periodontitis according to the area of the lesion, and gingivitis refers to a disease in which the lesion occurs only in the gum, that is, soft tissue, and periodontitis refers to the lesion. Refers to a condition that manifests itself in areas that extend to the gums and around the gum bone.

As used herein, the term "periodontal bacteria" refers to a causative agent causing periodontal disease by secreting virulent metabolites while infecting the alveolar bone or the tissues surrounding the alveolar bone, preferably gram-negative anaerobic bacteria. And more preferably Actinobacillus sp .) Microorganisms, Actinomyces sp .) microorganism, Campylobacter sp .) microorganisms, Enterococcus sp .) microorganism, Fusobacterium sp .) microorganisms, Klebsiella sp .) microorganisms, Pantoea sp .) microorganisms, Peptostreptococcus sp .) microorganisms, Porphyromonas sp .) microorganism, Staphylococcus sp .) microorganisms, Streptococcus sp .), Treponema sp .) microorganisms, Veillonella sp .) microorganisms, Propionibacterium sp . microorganisms, and the like, most preferably Enterococcus faecalis ), Actinomyces viscosus ), Streptococcus mutans ), Streptococcus sanguis ) and the like, but is not particularly limited thereto.

The term "prevention" of the present invention refers to all actions of inhibiting or delaying periodontal disease by administration of the gangjinhyang extract, nerolidol or its pharmaceutically acceptable salts isolated from the gangjinhyang extract.

The term "treatment" of the present invention is any action in which the symptoms of periodontal disease are improved or cured by administration of a composition comprising the Gangjin flavor extract, nerolidol isolated from the Gangjin flavor extract, or a pharmaceutically acceptable salt thereof. Means.

According to one embodiment of the present invention, the nerolidol isolated from hot water extract, 70% ethanol extract, or strong extract of Gangjin-hyang is Enterococcus pecalis, Actinomyces viscous, Streptococcus mutans and Streptococcus It may exhibit antimicrobial activity in a concentration-dependent manner with respect to Sangwis (Figs. 1 to 6). As described above, by confirming that the periodontal bacteria specifically exhibit a concentration-dependent antimicrobial activity, it was confirmed that the Kangjinhyang extract of the present invention or nerolidol isolated therefrom is effective for the prevention or treatment of periodontal disease.

In addition, the pharmaceutical composition comprising the strong extract according to the present invention may further comprise a suitable carrier, excipients and diluents commonly used in the preparation of the pharmaceutical composition. Examples of the carrier, excipient and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

On the other hand, the pharmaceutical compositions of the present invention, respectively, in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, oral formulations, external preparations, suppositories, and sterile injectable solutions, respectively, may be used according to conventional methods. Can be. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid form preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, which form at least one excipient such as starch, calcium carbonate, sucrose or It is prepared by mixing lactose and gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use may include various excipients such as wetting agents, sweetening agents, fragrances, preservatives, etc. in addition to water and liquid paraffin, which are simple diluents commonly used in suspension, liquid solutions, emulsions and syrups have. 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.

According to another embodiment, the present invention provides a food composition for preventing or ameliorating periodontal disease, including gangjinhyang extract, nerolidol or a pharmaceutically acceptable salt thereof isolated from the gangjinhyang extract.

Kangjinhyang extract of the present invention has been used as a medicinal herb in the past, and the nerolidol isolated from the Kangjinhyang extract can also be used as a food composition because it is derived from the extract of Gangjinhyang has been used as a medicinal herb from the prior .

The gangjinhyang extract, nerolidol or its pharmaceutically acceptable salts isolated from the gangjinhyang extract comprises 0.01 to 100% by weight, more preferably 1 to 80% by weight relative to the total weight of the food composition. When the food is a beverage, it may be contained in a proportion of 1 to 30 g, preferably 3 to 20 g based on 100 ml. In addition, the composition may include additional ingredients that are commonly used in food compositions to improve the smell, taste, time and the like. For example, it may include vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, panthotenic acid, and the like. In addition, minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu) and chromium (Cr) may be included. It may also contain amino acids such as lysine, tryptophan, cysteine, valine and the like. In addition, preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), fungicides (bleaching powder and highly bleaching powder, sodium hypochlorite, etc.), antioxidants (butylhydroxyanisol (BHA), butylhydroxytoluene ( BHT), etc.), colorants (such as tar pigments), colorants (such as sodium nitrite, sodium nitrite), bleach (sodium sulfite), seasonings (such as MSG glutamate), sweeteners (ducin, cyclate, saccharin, sodium, etc.) Food additives such as flavors (vanillin, lactones, etc.), swelling agents (alum, potassium D-tartrate, etc.), reinforcing agents, emulsifiers, thickeners (pigments), coatings, gum herbicides, foam inhibitors, solvents, improvers, etc. ) Can be added. The additive is selected according to the type of food and used in an appropriate amount.

On the other hand, by using a food composition comprising the gangjinhyang extract, nerolidol isolated from the gangjinhyang extract or a pharmaceutically acceptable salt thereof functional food for preventing or improving periodontal disease can be prepared.

As a concrete example, the above-mentioned composition can be used to produce a processed food having good shelf-life while modifying the characteristics of agricultural products, livestock products or aquatic products. Such processed foods include, for example, sweets, beverages, alcoholic beverages, fermented foods, canned foods, milk processed foods, land processed foods, noodles and the like. Confections include biscuits, pies, cakes, breads, candies, jelly, gum, cereals (including meal substitutes such as grain flour). Beverages include carbonated drinks, functional hot drinks, juices (eg, apples, pears, grapes, aloes, citrus fruits, peaches, carrots, tomato juices, etc.), Sikhye, and the like. The mainstream includes sake, whiskey, shochu, beer, liquor, and fruit wine. Fermented foods include soy sauce, miso, and kochujang. Canned food includes canned seafood (e.g., tuna, mackerel, saury, canned seashells, etc.), canned animal products (beef, pork, chicken, canned turkey, etc.), canned produce (corn, peaches, canned apples, etc.). Milk processed foods include cheese, butter, yogurt and the like. Meat processed foods are pork cutlet, beef cutlet, chicken cutlet, sausage. Includes sweet and sour pork, nuggets, breadfruits and more. And noodles such as sealed packaging raw noodles. In addition, the composition may be used in retort food, soup and the like.

The term "functional food " of the present invention is the same term as " food for special health use (FoSHU) ", and includes medical and medical effects It means high food. As used herein, the term "health food" refers to a food having an active health maintenance or promotion effect compared to a general food, and a health supplement food refers to a food for health supplement purposes. Accordingly, the terms functional foods, health foods, and health supplements are widely used.These foods may be prepared in various forms such as tablets, capsules, powders, granules, liquids, pills, etc. in order to obtain useful effects in the prevention or improvement of periodontal disease. Can be.

According to another embodiment, the present invention provides a composition for oral cleaning comprising gangjinhyang extract, nerolidol isolated from the gangjinhyang extract, or a pharmaceutically acceptable salt thereof as an active ingredient.

Gangjinhyang extract provided by the present invention, nerolidol isolated from the Kangjinhyang extract or a pharmaceutically acceptable salt thereof has the effect of inhibiting the growth of periodontal bacteria that can cause periodontal disease by infecting the periodontal of the oral cavity. Since the mouth is clean, it can be used as an effective ingredient of the composition for oral cleaning, which can prevent the development of various oral diseases including periodontal disease.

In addition, the composition for oral cleaning provided by the present invention may further include caries preventive agent, fungicide, herbal medicine, inflammation relieving agent, enzyme, etc. in addition to the active ingredient in order to improve the cleaning effect. At this time, the caries preventive agent is not particularly limited, but sodium fluoride, sodium fluoride phosphate, amine fluoride, tin fluoride, xylitol and the like can be used; The fungicide is not particularly limited thereto, and chlorohexidine, cetylpyridium chloride, triclosan and the like can be used; The herbal medicine is not particularly limited to this, but may include bamboo salt, propolis, and the like; Examples of the anti-inflammatory agent include, but are not limited to, allantoin, aminocaproic acid, tranexamic acid, tocopherol acetate, methylsulfonylmethane, and the like; Examples of enzymes include, but are not limited to, dextranase, glucose oxidase, glucoamylase, α, β-amylase, lactoperoxidase, lysozyme, and the like.

On the other hand, the composition for oral cleaning of the present invention may be formulated in the form of a solid, paste, powder, liquid or spray by adding an abrasive, a wetting agent, a binder, a foaming agent, etc. It may further include. At this time, the abrasive is not particularly limited, but precipitated silica, silica gel, zirconium silicate, calcium dihydrogen phosphate, calcium dihydrogen phosphate, hydrous alumina, hard calcium carbonate, heavy calcium carbonate, calcium pyrophosphate, insoluble methacrylate, aluminum Silicates and the like can be used; Examples of the humectant include, but are not limited to, concentrated glycerin at a concentration of 98%, aqueous sorbitol at a concentration of 70%, an aqueous solution of amorphous sorbitol at a concentration of 70%, xylitol, polyethylene glycol, propylene glycol, and the like; Examples of the binder include, but are not limited to, sodium carboxymethylcellulose, carrageenan, xanthan gum, and the like; Examples of the foaming agent include, but are not limited to, sodium lauryl sulfate, a copolymer of polyoxyethylene polyoxypropylene (poloxamer), polyoxyethylene hardened castor oil, polyoxyethylene sorbitan fatty acid ester, and the like.

Gangjinhyang extract of the present invention, the composition comprising the nerolidol or pharmaceutically acceptable salts thereof isolated from the Kangjinhyang extract can be used safely without side effects, showing excellent and specific antimicrobial activity against periodontal bacteria Therefore, it may be widely used for more effective prevention or treatment of periodontal disease.

Figure 1 is a graph showing the cell number reduction level of each periodontal bacteria by hot water extract of Gangjin-hyang.
Figure 2 is a graph showing the cell count reduction level of each periodontal bacteria by 70% ethanol extract of Kangjin-hyang.
Figure 3 is a graph showing the cell number reduction level of each periodontal bacteria caused by nerolidol.
Figure 4 is a photograph showing the minimum sterilization concentration of the hydrothermal extract of Kangjin-hyang on each periodontal bacteria.
Figure 5 is a photograph showing the minimum bactericidal concentration of the 70% ethanol extract of Gangjin flavor on each periodontal bacteria.
Figure 6 is a photograph showing the minimum sterilization concentration of nerolidol on each periodontal bacteria.

Hereinafter, the present invention will be described in more detail with reference to examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.

Example  One: Periodontal bacteria  About Gangjin Hyang  Antimicrobial Activity of Extracts

Example  1-1: Periodontal  culture

Known as periodontal bacteria Enterococcus faecalis (Enterococcus faecalis ) (KCTC 3206, ATCC 19433), Actinomyces viscosus ) (KCTC 9146, ATCC 15988), Streptococcus mutans ) (KCTC 3065, ATCC 25175) and Streptococcus sanguis ) (NCTC 9811) was primary cultured in Brain Heart Infusion (BHI) Broth medium (BHI, Difco, USA), and the primary cultures were subcultured in BHI solid medium containing agar.

Example  1-2: Gangjin Hyang Heat number  Preparation of extract

50 g of dried Kangjin-hyang was immersed in 1 liter of distilled water, hydrothermal extraction for 3 hours, filtered to obtain a liquid component, and then lyophilized to prepare a hydrothermal extract of Kangjin-yang.

Example  1-3: Gangjin Hyang  Preparation of 70% Ethanol Extract

50 g of dried Gangjin fragrance was immersed in 0.5 liter of 70% ethanol, sonicated at 25 ° C. for 1 hour, filtered to obtain a liquid component, and then the solvent was removed using a rotary concentrator under reduced pressure. 70% ethanol extract was prepared.

Example  1-4: Periodontal bacteria  About Gangjin Hyang  Antimicrobial Activity of Extracts

In order to confirm the proliferation inhibitory activity of the gingival fungus extract against periodontal bacteria, each periodontal bacteria passaged in Example 1-1 was incubated in BHI Broth medium, and the bacterial solution was diluted so that the absorbance value was 0.1 at 550 nm. Diluted bacterial solution was inoculated at 1% in fresh BHI Broth medium and 200 μl was dispensed into each well of a 96-well plate. Gangjinhyang hot water extract prepared in Example 1-2 and Kangjinhyang 70% ethanol extract prepared in Example 1-3 were added to each of the dispensed wells at a concentration of 2.5 mg / ml and incubated for 20 hours, After the incubation was completed, the absorbance value was measured at 550 nm to determine the growth inhibitory activity against periodontal bacteria (FIGS. 1 and 2).

1 is a graph showing the cell count reduction level of periodontal bacteria by hot water extract of Kangjin-hyang, Figure 2 is a graph showing the cell number reduction level of periodontal bacteria by 70% ethanol extract of Kangjin-hyang. As shown in Figure 1 and Figure 2, Kangjinhyang hot water extract and 70% ethanol extract can reduce the number of cells of periodontal bacteria in a concentration-dependent manner, strains that are most reduced by the Kangjinhyang extract is Streptococcus Mutans, followed by Streptococcus saint gux, Actinomyces viscous, and Enterococcus pecalis.

As such, different levels of antibacterial activity were shown for each periodontal bacteria by the Gangjinhyang extract, which was analyzed as the Kangjinhyang extract of the present invention exhibiting specificity and selectivity for periodontal bacteria.

Example  1-5: Periodontal bacteria  About Of nerolidol  Growth inhibitory activity

Instead of Gangjinhyang hot water extract or 70% ethanol extract, except for treating each well of a 96-well plate with nerolidol (sigma-aldrich), known as the ingredient contained in Kangjinhyang extract, at a concentration of 0.625 mg / ml Was carried out in the same manner as in Example 1-4, to determine the growth inhibitory activity against periodontal bacteria (Fig. 3).

Figure 3 is a graph showing the cell number reduction level of periodontal bacteria caused by nerolidol. As shown in Figure 3, nerolidol known to be included in the Gangjinhyang extract was able to reduce the number of cells of periodontal bacteria in a concentration-dependent. Although nerolidol appears to exhibit the same antimicrobial activity as Kangjin-hyang's hydrothermal extract in that it reduces the number of cells of Streptococcus mutans most, and reduces the number of cells of Enterococcus pecalis the least, it has been shown that Antibacterial activity distinguished from each Kangjin-hyang extract in that it can reduce the number of cells of Actinomyces biscusus more than Gus, and can reduce the number of cells of Enterococcus pecalis even by low concentration. It could be seen that.

However, since nerolidol showed different levels of antibacterial activity for each periodontal bacteria, nerolidol was also analyzed to show specificity and selectivity for periodontal bacteria similarly to Kangjin flavor extract.

Example  2: Periodontal bacteria  Determination of Minimum Sterilization Concentration

2 μl of each culture cultured in Examples 1-4 and 1-5 was inoculated into BHI solid medium used in Example 1-1, and cultured to determine minimum bactericidal concentration (MBC) ( 4 to 6).

Figure 4 is a photograph showing the minimum bactericidal concentration of Kangjin-hyang hot water extract on periodontal bacteria, Figure 5 is a photograph showing the minimum bactericidal concentration of 70% ethanol extract of Kangjin-flavor on periodontal bacteria, Figure 6 The photograph shows the minimum bactericidal concentration of nerolidol.

First, in the case of hot water extract of Kangjin-hyang, the minimum sterilization concentration for Enterococcus pecalis, Actinomyces biscusus and Streptococcus guigus is more than 2,500 µg / ml, and the minimum sterilization concentration for Streptococcus mutans is It was confirmed that it is 2,500 μg / ml (FIG. 4). Next, in the case of the 70% ethanol extract of Kangjin-hyang, the minimum bactericidal concentration for Enterococcus pecalis and Actinomyces viscous was 2,500 µg / ml or more, and the minimum bactericidal concentration for Streptococcus guigus was 2,500 µg. / Ml, the minimum sterilization concentration for Streptococcus mutans was confirmed to be 1,250 μg / ml (FIG. 5). Finally, for nerolidol, the minimum bactericidal concentrations for Enterococcus pecalis, Actinomyces viscous and Streptococcus guigus are more than 2,500 µg / ml and the minimum sterilization concentration for Streptococcus mutans is 1,250. It was confirmed that the μg / ㎖ (Fig. 6).

These results were analyzed in accordance with the results of Examples 1-4 and 1-5 in that the strain most affected by the antimicrobial activity of Kangjin-hyang extract and nerolidol was Streptococcus mutans.

Example  3: Gangjin Hyang  Extract Acute Toxicity Experiment

Example  3-1: Preparation of Laboratory Animals

Four-week-old male and female ICR mice were obtained and visually inspected for appearance, and then purified in an animal room where the test was conducted for one week, and healthy animals were selected for the experiment. During the acclimatization period, the general clinical symptoms were observed, and 20 healthy males and females were selected, and the experimental animals were prepared by performing group separation by random method according to the weight range. The experiment for this embodiment was deliberated by the Experimental Animal Ethics Committee, and during the period of acclimation and experiments, the temperature was 23 ± 3 ℃, relative humidity 50 ± 10%, the number of ventilation was 12-16 times per hour, and the illumination was The 12-hour contrast cycle (lighting up 7:00, lighting out at 19:00) and the illuminance were adjusted to 150-300 Lx to maintain a constant breeding environment. Solid animal feed (PMI nutrition, USA) and water were used as free intake conditions.

Example  3-: Gangjin Hyang  Extract Acute toxicity  evaluation

Using the experimental animals prepared in Example 3-1, the body weight of the experimental animals to which the Gangjinhyang hot water extract was administered was measured, and the acute toxicity due to the Kangjinhyang hot water extract administration was compared with the control group.

Specifically, three groups (five animals each) were prepared, each of which was set to azeotropic 0.5 based on the oral dose limit of 5000 mg / kg, and four male and female ICR mice were included, including the control group. Divided. The route of administration was the most widely used oral administration method in oriental clinic, and the herbal medicine of each dose group was disclosed in the experiment by diluting in saline immediately before administration, fasting the animal overnight, and fixing it by dorsal skin fixation and oral administration. Forced oral administration was performed using a metal soda and an injection tube.

Thereafter, while raising for 14 days (once a day), weight change was measured before and after 1, 3, 7, 14 days (Table 1), and whether or not a dead individual of the test animals occurred. It was confirmed (Table 2).

Changes in Body Weight of Experimental Animals Treated with Gangjinhyang Extract gender Strong Amount Dose (g / kg) Weight (g) 0 days 1 day 3 days 7 days 14 days cock 0
5
2.5
1.25 19.944
20.680
20.558
20.540 22.146
23.104
22.476
22.140 22.410
23.450
22.466
22.406 23.060
23.654
22.694
22.190 23.388
23.868
22.646
23.176 female 0
5
2.5
1.25 25.802
26.880
26.386
26.050 29.816
30.332
29.142
29.126 30.682
31.584
31.086
29.158 32.204
32.966
32.812
31.610 33.662
33.722
33.334
32.432

Number of deaths among experimental animals administered Gangjinhyang extract gender Strong Amount Dose (g / kg) The death population (mari) 0 One 2 3 4 5 6 7 8 9 10 11 12 13 14 cock 0
5
2.5
1.25 0
0
0
0 0
0
0
0 0
0
0
0 0
0
0
0 0
0
0
0 0
0
0
0 0
0
0
0 0
0
0
0 0
0
0
0 0
0
0
0 0
0
0
0 0
0
0
0 0
0
0
0 0
0
0
0 0
0
0
0 female 0
5
2.5
1.25 0
0
0
0 0
0
0
0 0
0
0
0 0
0
0
0 0
0
0
0 0
0
0
0 0
0
0
0 0
0
0
0 0
0
0
0 0
0
0
0 0
0
0
0 0
0
0
0 0
0
0
0 0
0
0
0 0
0
0
0

As shown in Table 1, no change in body weight of the experimental animals due to the administration of Gangjinhyang extract was observed. In addition, as shown in Table 2, no death of the experimental animals due to the administration of the extract of Kangjin-hyang. In particular, even when administered to 5g per unit body weight (1kg) of the experimental animals did not occur because the death of Kangjinhyang extract was confirmed that the LD ₅₀ value of at least 5g / kg or more.

Claims (13)

Pharmaceutical composition for the prevention or treatment of periodontal disease, including the Kangjin-hyang extract as an active ingredient.
The method of claim 1,
The strong jinhyang extract is a hydrothermal extract or 70% ethanol extract of the strong jinhyang.
The method of claim 1,
The periodontal disease is Enterococcus ( Enterococcus) faecalis ), Actinomyces viscosus ), Streptococcus mutans or Streptococcus sanguis is a disease caused by infection.
The method of claim 1,
The periodontal disease is periodontitis or gingivitis composition.
Pharmaceutical composition for the prevention or treatment of periodontal disease, including nerolidol or a pharmaceutically acceptable salt thereof as an active ingredient.
Food composition for the prevention or improvement of periodontal disease, including Kangjin flavor extract.
The method according to claim 6,
The strong jinhyang extract is a hydrothermal extract or 70% ethanol extract of the strong jinhyang.
The method according to claim 6,
The periodontal disease is a disease caused by infection of Enterococcus pecalis, Actinomyces biscusus, Streptococcus mutans or Streptococcus guigus.
The method according to claim 6,
The periodontal disease is periodontitis or gingivitis composition.
A food composition for preventing or ameliorating periodontal disease, including nerolidol or a pharmaceutically acceptable salt thereof.
Oral cleansing composition comprising the Kangjin flavor as an active ingredient.
12. The method of claim 11,
The strong jinhyang extract is a hydrothermal extract or 70% ethanol extract of the strong jinhyang.
Oral doldol or a pharmaceutically acceptable salt thereof as an active ingredient comprising a composition for oral cleaning.

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