KR100330924B1 - Oral composition containing extract of ulmi cortex for treatment of periodontal diseases - Google Patents

Abstract

PURPOSE: Provided is an oral composition containing Ulmi Cortex extract, Achyranthes japonica extract or the mixture thereof for treatment of periodontal diseases. It inhibits the generation of superoxide, PGE2, interleukin-1 beta, as well as collagenase, while promoting the synthesis of collagen protein. CONSTITUTION: An oral composition for treatment of periodontal diseases is characterized by containing Ulmi Cortex extract which is obtained by using an extract solvent selected from the group consisting of water, methanol, ethanol, propanol and butanol, in the amount of 0.01-5 wt.%. It further contains Achyranthes japonica extract.

Description

Composition for periodontal disease containing milk extract

The present invention relates to a composition for oral cavity containing hyssop or milky skin extract. More specifically, the present invention relates to a composition for oral cavity containing at least one of the extract of dew or baekbaekpi excellent in treatment effect against periodontal disease, extracted from dew or baekbaekpi as an active ingredient. As an example of the composition of this invention, a toothpaste composition and an ointment composition are mentioned.

Periodontal disease is clinically caused by loss of teeth due to gingival inflammation and bleeding, periodontal pocket formation and destruction of alveolar bone. This periodontal disease is a series of processes consisting of colonization of bacteria, infiltration of periodontal tissues of bacteria, and destruction of periodontal tissues. First, saliva proteins in oral saliva are adsorbed to dentin and chalky surfaces to form a film. Bacteria, such as Streptococcus and Actinomyces, grow on the surface and form plaques, and as time passes, these plaques move in the direction of the root canal, while at the same time, porphyromonas and actinos. Anaerobic Gram-negative bacteria such as Bacillus (Actinobacillus) grow, and these bacteria, bacterial components, and bacterial products penetrate into the gingival connective tissue through the gingival epithelium, forming periodontal sacs, and metabolism of these bacteria results in poisonous hydrogen sulfide and ammonia Cell walls while releasing toxins from cells such as amines Active oxygen, prostaglandins secreted outside the cell by various actions of humoral and cellular immune systems that are directly destroyed by endotoxins such as lipopolysaccharide, or stimulating the bioimmune system Inflammation of the gums by various types of cytokines such as prostaglandins, leukotriens, histamine, interleukins and the like, and periodontal by enzymes such as collagenase secreted from bacteria and leukocytes When collagen, a tissue matrix, is broken down, gum retraction occurs, and it is left unattended, it progresses to periodontal disease.

Chlorhexidine gluconate, cetylpyridium chloride, sanguinarine and triclosan, which can kill and kill periodontal disease bacteria in a short time in an effort to prevent the occurrence of periodontal disease Antibacterial agents and anti-inflammatory agents such as triamcinolone acetonide have been developed and applied to oral products such as dentifrice, toothpaste and ointment, but the occurrence of periodontal disease has not been fundamentally prevented yet.

Therefore, in recent years, efforts to suppress periodontal disease using herbal extracts such as myrrh, baekbaekpi, horse riding, green tea, licorice, gold, pogongyoung, gold and silver, and corn have been active in Korea, but they play a decisive role in causing periodontal disease. Plant extracts that inhibit the production of prostaglandins or inhibit the activity of collagenase that degrades periodontal tissues have not been identified, and in most cases, they inhibit plaque formation or promote antibacterial, anti-inflammatory, astringent, hemostatic, and blood circulation. Through the action of herbal medicines such as to prevent the occurrence of periodontal disease.

Until now, the most widely used method for treating periodontal disease is to use pharmacologically active ingredients that inhibit the activity of collagenase enzymes that break down periodontal tissues or inhibit the production of prostaglandins, which cause periodontal disease.

As an example of such a method, U.S. Patent No. 5230895 has developed a system in which a drug containing tetracycline is continuously delivered to treat periodontal disease by inhibiting the enzyme activity of collagenase that degrades periodontal tissue. European Patent No. 528468 A1 proposes a method for treating periodontal disease by inhibiting periodontal disease by inhibiting the production of prostaglandins, which are the periodontal disease-causing agents, with toothpaste containing triclosan and oral cleansers. It is.

However, the above-mentioned pharmacologically active ingredients do not have both effects of inhibiting the generation of periodontal disease-causing agents and at the same time inhibiting periodontal tissue degrading enzyme activity in the treatment of periodontal disease. In addition to the limitations of treatment, since it is a synthetic material, there are many side effects of long-term use. In particular, tetracycline is usually administered 250mg (1 tablet) four times a day to administer 1000mg each day for 5-7 days. In general, tetracycline has been reported to be clinically and bacteriologically improved, and the results show that tetracycline is significantly effective after a tartar removal. Research reports on long-term low-dose administration showed that tetracycline was administered 1000 mg / day for 2 weeks, followed by 250 mg once daily for 48 weeks to reduce clinically gingivitis and motility strains in subgingival plaque And loss of periodontal pocket depth or adhesion was reported. However, long-term use of low concentrations of tetracycline often results in the development of resistance to Gram-negative bacteria, and the expression of such resistant strains often occurs even when the antibiotics are stopped. Side effects should be very careful, such as vomiting, pain in the gastrointestinal tract, Diarrhea and tooth coloring occur very often, and there are limitations that pregnant women and young children should refrain from.

Therefore, the present inventors induced collagenase activity measurement method and periodontal disease by decomposing periodontal tissues in various herbal medicines and plant extracts in order to find a substance that shows excellent periodontal disease treatment effect with little side effects or toxicity even after long-term use. By using scientific methods such as quantification of interleukin-1β and prostaglandin (PGE ₂ ), superoxide production assay, collagen synthesis quantification method, Herbal and plant extracts were screened for promoting collagen synthesis. As a result, specific extracts of hyssop and milky skin inhibit the production of periodontal disease-causing agents and inhibit the activity of periodontal tissue degrading enzymes as described below. Promote collagen synthesis Leading to the confirmation that this is effective, and have completed the present invention.

Accordingly, it is an object of the present invention to provide a composition for oral cavity containing the extract of horseradish, milky skin extract or a mixture thereof.

The herbal extract used as an active ingredient in the composition for periodontal disease of the present invention is extracted from dew or milky skin.

One of the effective herbal medicines of the present invention is a yellowish gray, which is dried in the sun after removing the beard roots by digging roots of Achyranthes japonica, which is a plant of amaranth and plants distributed in all parts of Korea, Japan, and South Yellow River in China. Or yellowish brown stick or slightly curved, 15-90cm long, 3-7cm in diameter, cylindrical, with many longitudinal wrinkles and marks of side roots. The plane is grayish white or light brown, and the neck of the center is yellowish white, and the quality is hard, but it is easily broken. It is almost odorless and tastes slightly sweet and mucus. In the cross section under a microscope, the skin and the neck are distinguished by a clear formation layer. At the center of the neck is a small protozoa, in the flow cell containing the matter of calcium hydroxide and starch granules are not visible. Its pharmacological effects are known to be effective in diuretic, painful, arthritis, etc. The main ingredients are saponin, oleanolic glycoside, insect transformation hormones (inokosterone, ecdysterone), and betaine hydrate (betaine hydrate). ) And the like.

Yubaekpi, another effective herb used in the composition of the present invention, is an elmaceae plant that is distributed in North Korea, China, and Japan, Ulmus macrocapra, Ulmus pumila, and Elm davidiana. The root portion of the stem or root portion of the root is taken and dried in the sun, and pharmacological action is known to be diuretic, small seedling, canned, species, single action, the main components are beta-sitosterol (β-sitosterol), phytosterol and It consists of tannins. The term "milky skin" is also called "Yu Root Skin" or "hundreds of skin" depending on the region in some countries, but the usual "milk white skin" is more commonly used.

In the present invention, water or alcohol extracts are used as extracts of dew and baekbaekpi. The extract of dew and milk white skin used in the present invention is obtained by drying the dried dew and milk white skin, crushing and then pulverizing, extracting with water or alcohol, and filtering and concentrating the filtrate under reduced pressure.

Alcohol extraction solvents that can be used for extraction of dew and milky skin include lower alcohol solvents having 1 to 4 carbon atoms such as methanol, ethanol, propanol, butanol, and the like, and preferably ethanol is used.

The herbal extract of the present invention is contained in a dentifrice composition, an ointment composition or an oral composition exhibits the prevention and treatment of periodontal disease. The composition of the present invention may contain milk extract and hyssop extract in periodontal disease prevention or therapeutically effective amounts. The specific composition of the composition according to the present invention can be determined by those skilled in the art in consideration of the formulation, site of use, and the like. For example, when the composition is a toothpaste composition or an ointment composition, it is appropriate to use the ratio of 0.001 to 5% by weight, preferably 0.01 to 3% by weight based on the total weight of the composition of the milky skin extract, and the present invention In the toothpaste composition or ointment of the composition, even when mixing the extract of the dew and milky skin, each extract is preferably present in the same ratio as described above, when the two herbal extracts are combined based on the total weight of the composition It may be 0.002 to 10% by weight.

The composition for periodontal disease according to the present invention can be administered in oral tablets such as tablets, sugar tablets, powders, and capsules in suitable dosage forms, or oral compositions such as toothpaste compositions, mouthwashes and ointments for oral cavity. It may also be prepared by mixing with one or more excipients or additives (eg, surfactants, solubilizers, wetting agents, preservatives, fragrances, diluents, fillers, emulsifiers, lubricants, colorants, etc.) that are acceptable in compositions for preventing and treating periodontal disease. .

Specifically, the toothpaste composition according to the present invention can be prepared by the following method.

As the abrasive component, one or two or more selected from potassium monohydrogen phosphate, precipitated silica, silica gel, sodium bicarbonate, calcium carbonate, hydrous alumina, insoluble sodium phosphate and sodium pyrophosphate is used. The abrasive component is usually 1-90% by weight, but in the present invention, 20-60% by weight of calcium monohydrogen phosphate, hydrous silica and calcium carbonate alone or in combination.

As a fluorine compound which is an agent that promotes remineralization of teeth and strengthens the tissues of teeth, sodium fluoride or sodium monofluorophosphate is used alone or in combination of two or more thereof, and its content is suitably 0.01 to 2.0% by weight. If the content of the drug is less than 0.01% by weight is not sufficient effect and exceeds 2.0% by weight it is not suitable for oral preparations because it may affect the safety of the human body.

Wetting agents are used to maintain the dentifrice composition and prevent drying. Such wetting agents include glycerin, sorbitol solution, polyethylene glycol, and propylene glycol, which are used alone or in combination of two or more of them by 20 to 60 wt%.

In addition, binders are used to combine the liquid and solid components of the toothpaste component to maintain the form of the toothpaste and to ensure safety. Such binders are mainly sodium carginate or calcium salt, carboxymethyl cellulose sodium, xanthan gum, acacia gum, etc. Natural or synthetic polymeric materials, the content of which is from 0.1 to 5% by weight.

The foaming agent complements the cleaning action of the abrasive and not only penetrates the medicinal agent to the area that is difficult to reach the toothbrush, but also generates bubbles to increase the sensation of teeth, helps the cleaning action, speeds up the dispersion and penetration of the medicinal agent, It reduces the interfacial tension, so foreign substances in the oral cavity easily fall. Commonly used foaming agents include sodium lauryl sulfate, sodium alkyl sulfate, and anionic surfactants such as polyoxyethylene, polyoxypropylene, copolymers, polyoxyethylene hardened castor oil, polyoxyethylene sorbitan fatty acid, Alkanolamide fatty acid esters, sucrose fatty acid esters polyoxyethylene sorbitan fatty acid esters, polyoxyethylene fatty acid esters and polyoxyethylene castor hardened oil derivatives are used. The content of the foaming agent is preferably used 0.5 to 5% by weight of anionic or nonionic surfactants alone or in combination of two or more. In addition, flavors and sweeteners are used to control the hot or somewhat bitter taste, and the flavors are mainly used natural peppermint and spearmint oil. Perfume is used in the toothpaste 0.1 to 1% by weight. The sweetening agent is mainly used synthetic or natural non-fermentable sugars, and the representative ones are saccharin sodium, aspartame, lactose, maltose, ziitol, and the like, and suitable sweeteners are 0.05 to 1% by weight of saccharin sweetener. It is preferable.

In addition, buffers for adjusting the pH of the toothpaste composition include alkali metal salts of phosphate, in particular sodium phosphate, dibasic sodium phosphate, trisodium phosphate, citric acid and sodium citrate, phosphoric acid, hydrochloric acid, sodium hydroxide and sodium pyrophosphate and pyrophosphate. There is a buffer composition in the toothpaste composition according to the present invention is used by mixing two kinds of sodium phosphate, sodium phosphate, sodium phosphate, sodium phosphate, the pH is adjusted to 5 to 8.0. In order to prevent the contamination of microorganisms that may occur during the manufacture and use of toothpaste compositions, methyl paraoxybenzoate, benzoic acid, sodium benzoate, salicylic acid, etc., which are generally approved for use in foods and medicines, alone or in combination of two or more 0.01 to 0.5% by weight was used. In addition, in the toothpaste composition of the present invention, the content of mixed with the extract of dew and milky skin is preferably 0.002 to 10% by weight.

Ointment compositions of the present invention also include additives commonly used in the field of ointment preparation, for example surfactants, solubilizers, wetting agents, drug delivery systems for oral soft tissues, buffering agents, preservatives, sweeteners and flavoring agents to stabilize the condition of the ointment. Components such as these may be further blended.

In the ointment composition of the present invention, as the surfactant for the state stability of the ointment, a polyoxyethylene-polyoxypropylene copolymer is used, such as Pluronic F-127 or Pluronic F-108. Pluronic derivatives can be used. In the ointment composition of the present invention such a surfactant can be used in combination in a ratio of 5 to 30% by weight based on the total weight of the composition.

In the ointment composition of the present invention, a lower alcohol solvent may be used as a solubilizer for extracts of dew and milky skin. Lower alcohol solvents used for this purpose include ethanol, isopropyl alcohol, etc., may be used by mixing two or more alcohol solvents. The lower alcohol solvent in the composition of the present invention is preferably blended in a ratio of 1 to 20% based on the total weight of the ointment composition.

The ointment composition of the present invention also uses a wetting agent to maintain the state of the ointment and prevent drying, such wetting agents include polyethylene glycol, such as glycerin, sorbitol liquid, polyethylene glycol 200, polyethylene glycol 400, polyethylene glycol 600 and polyethylene glycol 1000, One component selected from propylene glycol, poloxamer 407 and monoglyceride (Myverol 18-99) may be used alone, or two or more components may be mixed and used. The amount of wetting agent is preferably 5 to 40% by weight based on the total weight of the composition.

On the other hand, as a drug delivery system that can be delivered to the oral soft tissue to deliver the efficacy of the active ingredient gelatin and pectin alone or in combination to use in a ratio of 1 to 30% by weight based on the total weight of the composition, ointment Buffers to adjust the pH of the composition are selected from alkali metal salts of fixed phosphoric acid, in particular, sodium phosphate dibasic, sodium phosphate dibasic, sodium triphosphate, citric acid and sodium citrate, phosphoric acid, hydrochloric acid, sodium hydroxide and sodium pyrophosphate and pyrophosphate One type or two or more types of buffers may be used. In the ointment composition of the present invention, a pH value of about 5.0 to 8.0 may be obtained by appropriately mixing two kinds of sodium phosphate, sodium phosphate, and sodium phosphate. It is desirable to adjust as much as possible.

In addition, flavors and sweeteners are used to control the hot or somewhat bitter taste, considering that the composition of the present invention is especially applied in the oral cavity, which is generally a natural flavor peppermint or spearmint oil based on the total weight of the composition It is used at a ratio of 0.1 to 1% by weight. As the sweetening agent, synthetic or natural non-fermentable sugars are generally used. Representative examples of sweeteners that can be used include saccharin sodium, aspartame, lactose, maltose, xyitol, and the like, in particular saccharin sodium. The sweetener in the composition of the present invention is preferably used in a ratio of 0.05 to 1% by weight based on the total weight of the composition.

The ointment compositions of the present invention also contain preservatives, such as methyl paraoxybenzoate and paraoxybenzoate, which are generally allowed for use in foods and pharmaceuticals to prevent contamination of microorganisms that may occur during the manufacture and use of ointments. , One or two or more components selected from the group consisting of benzoic acid, sodium benzoate, salicylic acid and the like are used in a ratio of 0.01 to 0.5% by weight based on the total weight of the composition.

When using the ointment composition of the present invention clinically, it is suitable to use the composition formulated in the above ratio so that 50-500mg, preferably 100-300mg is applied to the affected area with periodontal disease per day. The periodontal disease may be appropriately increased or decreased in consideration of factors such as the severity of the disease and the size of the affected part.

The invention is explained in more detail based on the following examples and comparative examples, but the invention is not limited in any way by these.

Preparation Example 1

50 g of powder obtained by pulverizing dried dew drops was added, and 500 ml of 95% ethanol was added thereto, followed by immersion extraction for 3 days, followed by filtration using No. 1 Whatman No. 1, and then concentrated under reduced pressure to dry. 15 g of dew extract was obtained.

Preparation Example 2

50 g of the powder obtained by pulverizing the dried milky skin was taken, and 500 ml of 95% ethanol was added thereto, followed by immersion extraction for 3 days, filtered using No. 1 Whatman No. 1, and then concentrated under reduced pressure to dry. 20 g of the obtained milky skin extract was obtained.

Example 1-18 and Comparative Example 1-18

Toothpaste compositions of Examples and Comparative Examples were prepared according to the compositional ratios shown in Tables 1 to 9 below.

TABLE 1

TABLE 2

TABLE 3

TABLE 4

TABLE 5

TABLE 6

TABLE 7

TABLE 8

TABLE 9

Experimental Example 1-1. Inhibitory Effect on Superoxide Production of Toothpaste (Examples 1-1 to 1-18)

The venous blood collected from citric acid as an anticoagulant was centrifuged at 1200 rpm for 10 minutes from a healthy adult without systemic disease, and then the leukocyte concentrate was recovered first and then diluted 1: 1 with RPMI 1640 medium. After adding 12 ml of Ficoll-Paque to 50 ml centrifuge tube, 30 ml of diluted blood was carefully added to form a middle layer, followed by centrifugation at 1600 rpm for 30 minutes, and then the upper layer containing serum was removed and the monolayer containing mononuclear cells. After careful dilution, add 3 times RPML 1640 medium and centrifuge at 800 rpm for 10 minutes, discard the supernatant, add 10 ml of RPMI 1640 medium, gently pipet, and centrifuge at 800 rpm for 10 minutes. the supernatant was discarded while the HBSS (hanks' balanced salt solution) was added to a buffer solution in 10 Pipetting the mononuclear white blood cells of a person in a 24-well freight ⁶ After dispensing 0.45 ml into cells / well and incubating for 2 hours aseptically under 95% air, 5% CO ₂ and 100% humidity conditions, FMLP (N-Formyl-Met-Leu-Phe) was added to 10 ^-6 M. After 0.05ml treatment and incubation for 15 minutes at 37 ℃ to stimulate the cells, 0.1μl cytochrome C (80μM), 0.1ml Superoxide dismutase (30μg), cellular interface 0.1 ml of the diluted toothpaste composition of Example and Comparative Example except for the active agent was added, and HBSS (hanks' bala-nced salt solution) was added so that the total reaction solution was 0.9 ml. 0.1 ml of the phylated Zymosan A, a substance, was added to a final concentration of 1.3 mg / ml, and kept at 37 ° C for 90 minutes with shaking, followed by 10 minutes at 4 ° C to stop the reaction, and then 4 ° C. After centrifugation at 1500 rpm for 10 min, the supernatant was measured at 550 nm for optical density and superoxide an The amount of ion produced is calculated by the following equation.

△ O.D. = (B-D)-(A-C) = (B + C)-(D + A)

The measured results are shown in Table 10 below.

TABLE 10

In the above test results, in Example 1-1, 5, 7, 9, and 11 containing more than 0.001% of hyssop extract, the inhibitory effect of superoxide production was increased with increasing concentration of hyssop extract, whereas Examples 1-2, 4, 6, 8, 10 and 12 containing the extract was found to have no superoxide inhibitory effect. Examples 1-13, 14, 15, 16, 17, and 18, which contain both dew extract and milk extract, also showed that the inhibitory effect on superoxide production increased with the concentration of dew extract.

Experimental Example 1-2. Inhibitory Effect on Collagenase, a Periodontal Tissue Degrading Enzyme of the Toothpaste of the Invention (Examples 1-1 to 1-18)

Experimental results of the present invention with the toothpaste inhibitory effect on collagenase, a periodontal tissue degrading enzyme is as follows.

In the course of this procedure, the periodontal pathogens Porphyromonas gingivalis and Polymorphonuclear It mimics the bio-oral environment in which periodontal tissue collagen is degraded by the action of collagenase enzymes in the saliva and gingival fluid of periodontal disease secreted from leukocytes) and neutrophils. Experimental method.

100 μl of Azocoll solution, 2% red collagen substrate, was added to 25 1.5 ml Eppendorf tubes, and one eppendorf tube was used as a blank and three tubes were purchased from Sigma. Sepaacryl from the saliva and gingival sulcus of the patient was added to the collagenase type I standard enzyme solution (collagen degrading activity: 315 units / mg) to 10, 100, and 200 ppm of each other 100 μl of collagenase enzyme purely separated through S-200 chromatography was used as a control and another tube was used as a control group, except for a cytotoxic surfactant in each of the remaining tubes (Examples 1-1 to 1). 1-18) and Comparative Group Toothpaste (Comparative Examples 1-1 to 1-18) were mixed with distilled water at a ratio of 1: 2, completely homogenized, and centrifuged at 5000 g for 10 minutes to treat 10 μl of the supernatant, followed by buffer solution (0.05 MTris-HCl, 1 nM CaCl ₂ , 7.8) was added to 500 μl of the reaction solution and reacted at 37 ° C. for 18 hours. Centrifugation of the eppendorf tube at 10000 g for 5 minutes precipitated undigested collagen and the supernatant containing the degraded collagen The absorbance was measured at 540 nm to prepare a standard activity curve, and the enzyme activity concentration was converted from the standard curve to compare and evaluate the enzyme activity of the experimental and control groups to obtain the following results (see Table 11 and Table 12).

TABLE 11

TABLE 12

In the above test results, the inhibitory effect on collagenase activity in Examples 1-1, 5, 7, 9, and 11 containing 0.01% or more of hyssop extract was found to increase as the concentration of hyssop extract increased. In Examples 1-2, 4, 6, 8, 10, and 12 containing milk extract, the inhibitory effect on collagenase activity was shown to increase as the concentration of milk extract was increased. Both extracts showed excellent inhibitory effects on collagenase activity and synergistic effects were found in Examples 1-13, 14, 15, 16, 17 and 18 containing both hyssop and milky skin extracts.

Experimental Example 1-3. Inhibitory Effect of Toothpaste of the Invention (Examples 1-1 to 1-18) on Interleukin (1L-1β) Production in Mononuclear Leukocytes

Blood mononuclear leukocytes isolated from blood were added to 0.8-well plate in a 24-well plate and divided into 10 ⁶ cells / well, and 200 μl of RPMI 1640 medium was added to the control and 100 μl of E. coli LPS (250 ppm). 100 μl of 3-fold diluted toothpaste composition of Examples (1-1 to 1-18) and Comparative Examples (1-1 to 1-18), except 100 μl of Well and LPS (250 ppm) and a surfactant with cytotoxicity After culturing for 24 hours using the well added as an experimental group, 50 μl of arachidonic acid was added and further incubated for 30 minutes. To a well of a 96-well plate to which an antibody of interleukin (1L-1β) was attached, 50 μl of standard solution (0, 10.24, 25.6, 64, 160, 400 pg / well) was added, and then the experimental group wells. 50 μl of the above cell culture solution was added, 50 μl of Biotinylated Antibody Reagent was added to all wells, and maintained at 25 ° C. for 30 minutes, followed by three washes with a washing buffer solution. Streptavidin-HRP Conjugate was added to all wells. After the addition, the mixture was maintained at 25 ° C. for 30 minutes, washed again with washing buffer solution three times, and 100 μl of enzyme substrate was immediately added thereto, and kept at 25 ° C. in the dark for 30 minutes with the lid of the plate open. After adding 100 μl, absorbance at 450 nm was measured with a microplate reader, and a standard curve was prepared using the absorbance values of the standard solution.

The measurement results are shown in Table 13 below.

TABLE 13

Inhibitory effects on 1-3, 5, 7, 9 and 11 containing E. coli LPS-stimulated human mononuclear leukocytes on the production and effect of 1L-1 on 0.01% concentration Was found to increase with increasing concentrations of dew extract, while Examples 1-2, 4, 6, 8, 10 and 12 containing milk extract showed no inhibitory effect on the production of 1L-1β. . Examples 1-13, 14, 15, 16, 17, and 18, which contain both dew extract and milk extract, also showed that the inhibitory effect on the production of superoxide was increased according to the concentration of the dew extract.

Experimental Example 1-4. Inhibitory Effect of Toothpaste of the Invention (Examples 1-1 to 1-18) on Prostaglandin (PGE2) Production in Mononuclear Leukocytes

Experimental results of the present invention with the toothpaste inhibitory effect on the production of prostaglandin, a periodontal disease-causing agent is as follows.

This test method was performed by stimulating human mononuclear leukocytes with lipopolysacch-aride, a cell wall component of porphyromonas gingivalis, a periodontal disease agent during the periodontal disease progression. Production of dean (PGE ₂ ) was compared with experimental group toothpaste (Examples 1-1 to 1-18) except for cytotoxic surfactants by immuno-diagnosis of antigen-antibody (Comparative Examples 1-1 to 1-18) ) Was mixed with distilled water at a ratio of 1: 2, homogenized completely, and centrifuged at 5000 g for 10 minutes to treat supernatant, and then evaluated the inhibitory effect.

Specific test methods contained 50 μl buffer (0.9% NaCl, 0.1% bovine serum albumin, 0.5% Kathon) in a blank well of a 96-well plate to which 1 gG of goat anti-mouse was attached. 0.1 M phosphate buffer solution) was added to the standard wells (0, 2.5, 5, 10, 20, 40, 80, 160, 320 pg), and 50 μl of the standard solution was added as well. 50 μl of the experimental group toothpaste (Examples 1-1 to 1-18) and the comparative group toothpaste (Comparative Examples 1-1 to 1-18), except for the cytotoxic surfactant, were added to the wells determined in the Examples and Comparative Examples. adding an antibody to PGE ₂ in the 50㎕ 50㎕ of PGE ₂ conjugate (conjugate) peroxidase (peroxidase) was added to all wells except for the blank well to all wells except for the blank well and 96-well-a freight Cover and keep at 25 ° C. for 1 hour, then wash 4 times with washing buffer solution (phosphate buffer solution containing 0.05% Tween 20: pH7.5). Immediately add 150 μl of enzyme substrate (3, 3 ', 5, 5'-tetramethylbenzidine / hydrogen peroxide dissolved in 20% dimethylformamide) at room temperature, hold at 25 ° C. for 30 minutes, and add 100 μl of 1M sulfuric acid. After the addition, the absorbance was measured at 450 nm with a microplate reader, and a standard curve was prepared using the absorbance values of the standard solution to calculate the amount of prostaglandin produced in the experimental group. The measurement results are shown in Table 14 and Table 15 below.

TABLE 14

TABLE 15

E. coli LPS-stimulated human mononuclear leukocytes stimulated PGE ₂ production and resulted in inhibitory effect on 1-3, 5, 7, 9 and 11 containing hyssop extract. On the other hand, it was found to be elevated, whereas Examples 1-2, 4, 6, 8, 10 and 12 containing the milky skin extract did not have an effect of inhibiting PGE ₂ production. Examples 1-13, 14, 15, 16, 17 and 18, which contain both dew extract and milk extract, also show that the PGE ₂ production inhibitory effect increases with the concentration of the dew extract.

Experimental Example 1-5. Effect of Toothpaste of the Invention (Examples 1-1 to 1-18) on the Collagen Protein Production of Gingival Fibroblasts

Primary culture One gingival fibroblast was dispensed to 10 ⁶ cell / well in a 24-well plate and incubated in DMEM medium containing 10% FBS for one day, and then replaced with a fresh medium for 24 hours. Wash the wash layer on the bottom with HBSS buffer solution, add 0.8ml of MEM medium without serum and Proline, and then use the experimental group toothpaste except for the cytotoxic surfactant (Examples 1-1 to 1-18). Comparative toothpaste (Comparative Examples 1-1 to 1-18) was mixed with distilled water in a ratio of 1: 2, completely homogenized and centrifuged at 5000 g for 10 minutes to treat supernatant, followed by ¹⁴ C-Proline (10 Ci) 100 Cells were incubated with a culture solution containing μl. After 24 hours, total protein and collagen protein were measured. First, in order to measure the total amount of extracellular total protein, the culture medium of each well was placed in a sealed dialysis tube at one end and sealed at the other end of Cold Buffer (Tris-HCl 0.05mol / L, NaCl 0.2mol / L, CaCl). ₂ Dialysis with 0.05mol / L, phenylmethylsulfonylfluoride 0.3mN) for 24 hours, take 100µl each, put in Count-ing Vial, add 10ml Scintillation coctail, and irradiate with Liquid Scintillation Counter (LSC) for 1 minute. Was measured. To measure the total amount of intracellular total protein, 0.1N NaOH and 0.3mN of phenylmethylsulfonylfluoride were added to each well from which the cell culture medium was removed, and then maintained at 60 ° C. for 30 minutes to break down the cell membrane. Into the sealed dialysis tube and seal the other end with Cold Buffer (Tris-HCl 0.05mol / L, NaCl 0.2mol / L, CaCl ₂ 0.05mol / L, phenylmethylsulfonylfluoride 0.3mN). After completion of dialysis for 24 hours, 100 μl of each was added to the counting vial, and 10 ml of scintillation coctail was added to measure radioactivity for 1 minute by LSC. To measure the synthesis amount of intracellular and extracellular collagen protein, 100 μl of dialysis-treated cell culture solution and cell homogenate were respectively taken, and placed in 1.5 ml microtube. CollagenaseBuffer (0.05M Tris-HCl, 1mM CaCl _2, 0.03 mM phenylmethylsulphur) Polyvinylfluoride), 100 μl of collagenase enzyme (100 ppm), and then maintained at 37 ° C. for 3 hours to completely degrade collagen and then remove 1% of 50% trichloroacetic acid to remove undigested protein. And 500 µl of a solution containing tannic acid was added and then precipitated at 4 ° C. for 30 minutes, followed by centrifugation at 1000 × g for 5 minutes, and 100 µl of the supernatant was added to the counting vial, and 10 ml of scintillation coctail was added. Radioactivity was measured.

The measured results are shown in Table 16 below.

TABLE 16

In order to study the efficacy and effect on the collagen synthesis of human fibroblasts, Example 1-2 containing ¹⁴ C-Proline but containing the milky skin extract as a result of treating the dentifrice composition of the present invention in a cell culture solution as described above. , 4, 6, 8, 10 and 12 showed that collagen synthesis promoting effect was increased as the concentration of milky skin was increased, whereas in Examples 1-3, 5, 7, 9 and 11 which contained dew extract No collagen synthesis promoting effect was found. Examples 1-13, 14, 15, 16, 17, and 18, which contain both hyssop extract and milky white extract, also showed that the collagen synthesis promoting effect increased with the concentration of milky skin.

Experimental Example 1-6. Clinical experiment on efficacy and effect of the present invention toothpaste (Examples 1-1 to 1-18)

Results of conducting a clinical trial for the treatment of periodontal disease with the present invention toothpaste are as follows.

Subjects were evenly orthodontic and lacking tooth defects, and periodontal disease patients were selected from 30 to 50 years old by age of 30 at 30-year intervals. The clinical trials on the treatment effect of periodontal disease of the experimental group toothpaste (Examples 1-1 to 1-18) and the comparative group toothpaste (Comparative Examples 1-1 to 1-18) were performed by the following method.

First, the clinical experimental method of the toothpaste of the present invention was performed by the following method.

After dividing the test group into the test group and the comparison group, oral health education and correct brushing method were supplied, the same control toothbrush was supplied to all the test subjects, and the toothpaste was performed to score the initial gingivitis index and the test group and the comparative group toothpaste. After 1 week, 1 month, 3 months, and 6 months, oral examination was performed to check the gingivitis index. The gingivitis index was measured by inserting a cherry oriental probe into the gingival sulcus. After probing continuously around each tooth without applying force, and after 30 seconds, the bleeding state was measured and the score was recorded in the following manner to obtain a result.

The measurement results are shown in Table 17 below.

TABLE 17

In the above clinical trial results, 1L-1 and prostaglandin-induced periodontal disease, superoxide, and collagenase to inhibit the enzyme activity of collagenase, which degrades periodontal tissues, promote collagen synthesis, Treatment of periodontal disease of the toothpaste (Examples 1-1 to 1-12) of the present invention containing milk extract extracts capable of inhibiting enzymatic activity of genase was performed by extracting dew extract or milk extract from 1 week to 6 months. Its efficacy was superior to Comparative Examples (1 to 18), which did not contain, and the gingivitis index of Comparative Example continuously increased with time after one month, while using the toothpaste developed according to the present invention. The incidence of periodontal disease was suppressed and the gingivitis index was remarkably decreased over time. Examples 1-13, 14, 15, 16, 17, and 18 containing both were found to have a synergistic effect on inhibition of periodontal disease

Examples 2-1 to 2-18 and Comparative Examples 2-1 to 2-18

Based on the component combination ratios shown in Tables 18 to 26, each component was dissolved and gelled in a buffer according to a conventional ointment preparation method and a comparative composition (Examples 2-1 to 2-18) according to the present invention. Comparative Examples 2-1 to 2-18) were prepared.

TABLE 18

TABLE 19

TABLE 20

TABLE 21

Table 22

TABLE 23

TABLE 24

TABLE 25

TABLE 26

Experimental Example 2-1: Superoxide Formation Inhibitory Effect of Ointment of the Present Invention

The inhibitory effect of the ointment composition of the present invention on the production of superoxide, a periodontal disease-causing agent, was measured by the following method.

The venous blood collected from citric acid as an anticoagulant was centrifuged at 1200 rpm for 10 minutes from healthy adults without systemic diseases, and then the leukocyte concentrate was recovered first, and then diluted 1: 1 with RPMI 1640 medium. It was. 12 ml of Ficoll-Paque was added to a 50 ml centrifuge tube, and 30 ml of the diluted blood was carefully added to form a middle layer and centrifuged at 1600 rpm for 30 minutes. The supernatant containing serum was removed, the monolayer containing mononuclear cells was carefully extracted with a sterile pipette into a new centrifuge tube, and then 3 times of RPMI 1640 medium was added and centrifuged at 800 rpm for 10 minutes. Discard supernatant, add 10 ml of RPMI 1640 medium, gently pipette, centrifuge at 800 rpm for 10 minutes, discard supernatant, pipet with HBSS (Hank's Balanced Sal Solution) buffer, and The leukocytes were aliquoted into 0.45 ml of 10 ⁶ cells / well in a 24-well plate and incubated for 2 hours aseptically under 95% air, 5% CO ₂ , 100% humidity, and then FMLP (N-formyl- Met-Leu-Phe) was treated with 0.05 ml (10 ⁻⁶ M) and incubated at 37 ° C. for 15 minutes to stimulate the cells. 0.1 ml (80 µM) of cytochrome C, 0.1 ml (30 µg) of superoxide dismutase, and 0.1 ml of the ointment compositions of Examples and Comparative Examples diluted three-fold with HBSS were added thereto, followed by 0.9 ml of the total reaction solution. HBSS was added to keep warm at 37 ° C. for 10 minutes and phagocytized Zymosan A was added in an amount of 0.1 ml to give a final concentration of 1.3 mg / ml. After the reaction mixture was insulated for 90 minutes at 37 ° C. with shaking, the reaction mixture was placed in a refrigerator at 4 ° C. for 10 minutes to stop the reaction, and then centrifuged at 1500 rpm for 10 minutes at 4 ° C. The absorbance of the supernatant was measured at 550 nm, and the amount of superoxide anion produced was calculated by the following equation.

ΔO.D. = (B-D)-(A-C) = (B + C)-(D + A)

The measured results are shown in Table 27 below.

TABLE 27

In Example 2-1, 5, 7, 9, 11 containing more than 0.001% of the dew drops from the results shown in Table 27, the superoxide formation inhibitory effect was increased as the concentration of the dew drops, whereas milky Examples 2-2, 4, 6, 8, 10, and 12 containing blood showed no superoxide inhibitory effect. Example 2-13, 14, 15, 16, 17, and 18 containing both dew and milky skin also showed that the inhibitory effect on the production of superoxide was increased with the concentration of dew.

Experimental Example 2-2: Inhibitory effect on the activity of collagenase enzyme of the ointment of the present invention

The inhibitory effect of the ointment composition according to the present invention on collagenase, a periodontal tissue degrading enzyme, was tested by the following method.

This test method was performed in the course of periodontal disease, salivary disease of periodontal disease patients from periodontal disease-causing porphyromonas gingivalis, polymorphonuclear leukocytes, and neutral leukocytes (Neutrophil). It is an experimental method that mimics the bio-oral environment where the collagen, which is the substrate of periodontal tissue, is degraded by the action of collagenase enzyme contained in the gingival fissure solution.

Add 100 μl of Azocoll solution, 2% red collagen substrate, to each of 25 1.5 ml Eppendorf tubes, use one Eppendorf tube as blank and three tubes with Sigma Collagenase type I standard enzyme solution (collagen degradation activity: 315 units / mg) purchased from was added to 10, 100, 200 ppm. To each of the remaining tubes, 100 μl of collagenase enzyme isolated from the saliva and gingival fluid of the periodontal disease patient through Sephacryl S-200 chromatography was added, one of which was used as a control. Each of the remaining tubes was thoroughly homogenized by mixing experimental group ointments (Examples 2-1 to 2-18) and comparative group ointments (Comparative Examples 2-1 to 2-18) according to the present invention with distilled water in a ratio of 1: 2. 10 μl of the supernatant, which was centrifuged at 5000 g for 10 minutes, was added by a micropipette, and a buffer solution (0.05 M Tris-HCl, 1 nM CaCl ₂ , pH7.8) was added so that the total reaction solution was 500 μl. Each Eppendorf tube was placed in a 37 ° C thermostat and reacted for 18 hours, followed by centrifugation at 10000 g for 5 minutes to precipitate undigested collagen, taking a supernatant containing digested collagen, and measuring absorbance at 540 nm. Standard activity curves were prepared from the results obtained from the solution-added tubes, and the enzyme activities of the experimental and control groups were compared by comparing the activity concentrations of the enzymes in the remaining tubes from the standard curves. The measured results are shown in Table 28 below.

TABLE 28

From the results shown in Table 28, the inhibitory effect on collagenase activity in Examples 2-1, 5, 7, 9, and 11 containing more than 0.001% of the dew was increased as the concentration of the dew was increased. In Example 2-2, 4, 6, 8, 10, and 12 containing milky skin, the inhibitory effect on collagenase activity was increased as milky skin concentration was increased. Both of the blood showed an excellent inhibitory effect on collagenase activity, and in Examples 2-13, 14, 15, 16, 17, and 18 containing both whipped and milky skin, a synergistic effect was shown.

Experimental Example 2-3: Inhibitory effect of the ointment of the present invention on the production of interleukin (IL-1β) of mononuclear leukocytes

The inhibitory effect on the production of interleukin IL-1β, a periodontal disease-causing agent, of the ointment composition according to the present invention was measured by the following method.

In a 24-well plate, 0.8 ml of blood mononuclear leukocytes isolated from blood were added to a concentration of 10 ⁶ cells / well, and the wells to which 200 μl of RPMI 1640 medium was added were added to the control, E. coli. The wells to which 100 μl of E. coli LPS (250ppm) was added, and 100 μl of LPS (250ppm) and 100 μl of ointment diluted three times with RPMI 1640 medium were incubated in the experimental group for 24 hours, followed by 50 μl of arachidonic acid. Add and incubate for 30 more minutes. 50 μl of IL-1β standard solution (0, 10.24, 25.6, 64, 160, 400 pg / well) was added to the well of a 96-well plate to which the antibody of IL-1β was attached, and the cell culture solution was added to the experimental group well. Add 50 μl, add 50 μl of biotinylated antibody reagent to all wells, hold at 25 ° C. for 3 hours, then wash buffer solution (0.01 M phosphate buffer containing 0.05 wt% of Twin 20, pH). 7.5) three times. 100 μl of streptavidin-HRP conjugate was added to all wells and maintained at 25 ° C. for 30 minutes, and then washed three times with washing buffer, and 100 μl of enzyme substrate was immediately added thereto at 25 ° C. in the dark. Keep the plate open for 30 minutes, add 100 μl of 0.18M sulfuric acid, measure the absorbance at 450 nm using a microplate reader, and prepare a standard curve from the absorbance values of the standard solution. Interleukin production was calculated. The measured results are shown in Table 29 below.

TABLE 29

From the results set forth in Table 29 above, As a result of testing the inhibitory effect of the ointment of the present invention on the production of IL-1β in human mononuclear leukocytes stimulated with E. coli LPS, Example 2-3, 5, 7, 9, Inhibition effect in 11 was shown to increase with increasing concentration of whistle, whereas in Example 2-2, 4, 6, 8, 10, 12 containing milky skin, there was no inhibitory effect of IL-1β production. appear. Examples 2-13, 14, 15, 16, 17, and 18 containing both dew and milky skin also showed that the inhibitory effect on the production of IL-1β was increased with the concentration of the dew.

Experimental Example 2-4: Inhibitory effect on the production of prostaglandins (PGE ₂ ) of mononuclear leukocytes of the ointment of the present invention

The inhibitory effect of the ointment composition of the present invention on the production of prostaglandins, which is a periodontal disease-causing agent, was tested by the following method.

This test method uses lipopolysaccharide, a cell wall component of Porphyromonas gingivalis, a periodontal disease-causing organism, to stimulate the production of PGE ₂ by stimulating human mononuclear leukocytes. By immunodiagnostic method, the experimental group ointment (Examples 2-1 to 2-18) and the comparative group toothpaste and ointment (Comparative Examples 2-1 to 2-18) were mixed with distilled water at a ratio of 1: 2 and completely homogenized to obtain 10 minutes at 5000 g. After the supernatant was treated by centrifugation for a while, the inhibitory effect was evaluated. The specific experimental method is as follows.

To a blank well of a 96-well plate to which goat's anti-mouse IgG was attached, 50 μl buffer solution (0.1 M phosphate buffer containing 0.9% NaCl, 0.1% bovine serum albumin, 0.5% Kathon) was added. To the standard well, 50 μl of appropriate concentration (0, 2.5, 5, 10, 20, 40, 80, 160, 320 pg) of PGE ₂ was added to the standard solution, and the experimental group ointment prepared as described above (Example 2- 1 to 2-18) and 50 μl of comparative group ointment (Comparative Examples 2-1 to 2-18) were added to the wells set as Examples and Comparative Examples and 50 μl of antibody against PGE ₂ was added to all wells except the blank wells. After the addition, 50 μl of PGE ₂ conjugate / peroxidase was added to all wells except the blank wells, and covered with a 96-well plate and maintained at 25 ° C. for 1 hour, followed by washing buffer solution (0.05% Tween 20). Phosphate buffer solution: pH4.4), washed four times and at room temperature 3,3 dissolved in an enzyme substrate (20% dimethylformamide) 150 μl of ', 5,5'-tetramethylbenzidine / hydrogen peroxide) was added immediately, held at 25 ° C. for 30 minutes, and 100 μl of 1M sulfuric acid was added, followed by measurement of absorbance at 450 nm with a microplate reader. A standard curve was prepared from the values, and the prostaglandin production amount (unit pg) of the experimental group was calculated from the standard curve. The measured results are as described in Table 30 below.

TABLE 30

From the results set forth in Table 30 above, The inhibitory effect of the ointment composition of the present invention on the production of PGE ₂ in human mononuclear leukocytes stimulated with E. coli PLS was tested. As a result, the inhibitory effect was found in Examples 2-3, 5, 7, 9, and 11 containing E. coli PLS. It was found that the concentration was increased as the concentration of the dew was increased, whereas in Examples 2-2, 4, 6, 8, 10, and 12 containing milky skin, there was no effect of inhibiting PGE ₂ production. Examples 2-13, 14, 15, 16, 17, and 18 containing both dew and milky skin also showed that the PGE ₂ production inhibitory effect increased with the concentration of dew.

Experimental Example 2-5: Effect of the ointment of the present invention on collagen protein production of gingival fibroblasts

The effect of the ointment composition of the present invention on the production of collagen protein of gingival fibroblasts was measured by the following method.

To the primary cultured gingival fibroblasts in 24-well plates 10 ⁶ a 10% FBS-containing DMEM medium are 1㎖ so that the cells / well seeded in each well and cultured for one day, and then the medium replaced with fresh medium the day old After incubation for 24 hours, the cell layer attached to the bottom was washed with HBSS buffer solution, and then 0.8ml of MEM medium containing no serum and proline was added, and compared with the experimental group ointments (Examples 2-1 to 2-18). Group ointment (Comparative Examples 2-1 to 2-18) was diluted to a ratio of 1: 2 with DMEM medium, completely homogenized, and 100 μl of the supernatant obtained by centrifugation at 5000 g for 10 minutes was added, followed immediately by ¹⁴ C. Cells were incubated with a culture solution containing 100 μl of proline (10 μCi). After 24 hours the collagen protein was measured. First, in order to measure the amount of extracellular total protein, the culture medium of each well was placed in a sealed dialysis tube at one end, and the other end was sealed, followed by cold buffer (Tris-HCl 0.05 mol / l, NaCl 0.2 mol / l, CaCl). ₂ 0.05 mol / l, phenylmethylsulfonylfluoride 0.3mN) for 24 hours, and then 100 μl of each was added to a counting vial, and 10 ml of scintillation coctail was added to the liquid scintillation counter. Radioactivity was measured for 1 minute with Liquid Scintillation Counter (LSC). In order to measure the total amount of intracellular total protein, 0.1N NaOH and 0.3mN of phenylmethylsulfonylfluoride were added to each well from which the cell culture medium was removed, and then maintained at 60 ° C. for 30 minutes to destroy cell membranes. Into one sealed dialysis tube and sealed with the other end with cold buffer (tris-HCl 0.05 mol / l, NaCl 0.2 mol / l, CaCl ₂ 0.05 mol / l, phenylmethylsulfonylfluoride 0.3mN). After dialyzing for 24 hours, 100 μl of each was added to the counting vial, 10 ml of scintillation cocktail was added, and radioactivity was measured for 1 minute by LSC. In order to measure the total amount of intracellular and extracellular total collagen protein, 100 μl of dialysis-completed cell culture solution and cell homogenate were added to 1.5 ml microtubes, and the collagenase buffer solution (0.05 M Tris-HCl, 1 mM CaCl ₂ , 0.3 mM phenylmethylsulfonylfluoride) and 100 μl of collagenase enzyme (100 ppm) were added and then maintained at 37 ° C. for 3 hours to completely break down collagen and then remove 50% of triclone to remove undigested protein. After adding 500 µl of a solution containing roacetic acid and 1% tannic acid, precipitate for 30 minutes at 4 ° C, centrifugation at 1000xg for 5 minutes, take 100 µl of the supernatant, add 10 ml of scintillation cocktail to the counting vial, and add LSC. Radioactivity was measured for 1 min. The measured results are shown in Table 31 below.

Table 31

From the results described in Table 31 above, as a result of treating the ointment composition of the present invention to a cell culture solution containing ¹⁴ C-proline, Example 2-2, 4, 6, 8, 10, 12 containing the milky skin extract In the collagen synthesis promoting effect was shown to increase as the concentration of milky skin increased, whereas in Examples 2-3, 5, 7, 9, 11 containing the dew was not shown to promote collagen synthesis. Examples 2-13, 14, 15, 16, 17, and 18 containing both dew and milk white skin also showed that collagen synthesis promoting effect increased with the concentration of milk white skin.

Experimental Example 2-6: Treatment effect of periodontal disease of the ointment of the present invention (clinical experiment)

The therapeutic effect of the ointment of the present invention on periodontal disease was confirmed by the following clinical trial method.

After dividing the test subjects into the experimental group and the comparative group, the gingival smear was measured to measure the initial gingivitis index, and the experimental group ointment and the comparative group ointment were supplied at 200 mg once a day and 3 times a day after meals. Months later, an oral examination was performed to check the gingivitis index. In each case, the gingivitis index is measured by inserting a periodental probe into the gingival bulb and continuously probing around each tooth without applying force and measuring bleeding after 30 seconds. Results were obtained by recording scores on the basis as described. The measured results are shown in Table 33 below.

Table 32

Table 33

From the results shown in Table 33, it promotes the synthesis of moss and collagen, which can inhibit the production of IL-1β and prostaglandins and superoxide that cause periodontal disease, and inhibit the enzymatic activity of collagenase that degrades periodontal tissues. , The ointment (Examples 2-1 to 2-12) of the ointment of the present invention containing milky skin extract that can inhibit the enzymatic activity of collagenase treatment of the periodontal disease from 1 week to 1 month The efficacy was superior to Comparative Examples (2-1 to 2-18) containing no blood, and the gingivitis index in comparison was continuously increased with time after one month, The use of the developed ointment suppresses the development of periodontal disease, and the gingivitis index is remarkably decreased over time. In Examples 2-13, 14, 15, 16, 17, and 18, there was a synergistic effect on the suppression of periodontal disease.

Toothpaste composition and ointment composition according to the present invention not only inhibits the production of superoxide (prooxide), prostaglandin (PGE ₂ ), interleukin-1β (interleukin-1β) which is a periodontal disease-causing agent, collagen protein is a substrate of periodontal tissue The periodontal disease can be effectively treated by inhibiting the activity of collagenase enzyme that degrades the protein and promoting collagen protein synthesis.

Claims (14)

Composition for periodontal disease containing milk extract. The composition of claim 1, wherein the baekbaekpi extract is a baekpipi extract extracted with an extraction solvent selected from the group consisting of water, methanol, ethanol, propanol and butanol. The composition of claim 1 wherein the composition is a toothpaste composition, an ointment composition or an oral formulation. The composition of claim 1, wherein the composition further comprises a dew drop extract. 4. The composition of claim 3, wherein the dentifrice composition contains 0.01-5 wt% of milky skin extract. The composition of claim 3 or 5, wherein the toothpaste composition further comprises an abrasive selected from the group consisting of calcium monohydrogen phosphate, precipitated silica and calcium carbonate in an amount of 20 to 60 wt% based on the total weight of the composition. The composition of claim 3 or 5, wherein the dentifrice composition further contains sodium fluoride or sodium monofluorophosphate at 0.01 to 2 wt% based on the total weight of the composition. The composition of claim 3, wherein the ointment composition contains the milky skin extract in an amount of 0.001 to 5% by weight based on the total weight of the composition. 9. The ointment composition according to claim 3 or 8, wherein the ointment composition is a surfactant selected from nonionic surfactants and anionic surfactants, solubilizers selected from ethanol and isopropyl alcohol, glycerin, sorbitol liquid, polyethylene glycol 200, polyethylene glycol Drug delivery system selected from at least one wetting agent selected from 400, polyethylene glycol 600 and polyethylene glycol 1000, propylene glycol, poloxamer 407 and monoglycerides (Nyverol 18-99), gelatin and plastics, sodium phosphate primary At least one buffer selected from sodium diacid, sodium triphosphate, citric acid and sodium citrate, phosphoric acid, hydrochloric acid, sodium hydroxide and sodium pyrophosphate, methyl paraoxybenzoate, propyl paraoxybenzoate, benzoic acid, sodium benzoate, and salicylic acid At least one preservative, synthetic or natural ratio selected from among A composition further comprising a sweetener selected from fermentable sugars, and a fragrance. The composition of claim 3 or 8, wherein the ointment composition further comprises polyoxyethylene-polyoxypropylene copolymer at 5 to 30% by weight based on the total weight of the composition. The composition of claim 3 or 8, wherein the ointment composition further contains 1 to 20% by weight, based on the total weight of the composition, of a solubilizer selected from ethanol and isopropyl alcohol. According to claim 3 or 8, wherein the ointment composition is glycerin, sorbitol liquid, polyethylene glycol 200, polyethylene glycol 400, polyethylene glycol 600 and polyethylene glycol 1000, propylene glycol, poloxamer 407 and monoglycerides (Myverol 18 -99) further comprising 5 to 40% by weight, based on the total weight of the composition, of at least one humectant selected from the group consisting of: The composition of claim 3 or 8, wherein the ointment composition further contains 1 to 30% by weight, based on the total weight of the composition, of at least one component selected from the group consisting of gelatin and pectin. 9. The ointment composition according to claim 3 or 8, wherein the ointment composition is selected from the group consisting of first sodium phosphate, second sodium phosphate, third sodium phosphate, citric acid, sodium citrate, phosphoric acid, hydrochloric acid, sodium hydroxide, sodium pyrophosphate and pyrophosphate. Wherein the pH value is adjusted to between about 5.0 and 8.0 by combining at least one buffer selected.