KR0176015B1 - Periodontal disease prevention and treatment composition - Google Patents

Abstract

The present invention relates to a periodontal disease prevention and treatment composition comprising the jujube extract and the thick extract as an active ingredient. The jujube extract and the thick extract extract-containing composition according to the present invention have a specific compounding ratio, i.e., a compounding ratio of 1: 6 to 1: 12, and particularly 1: 8 to 1: 10, Not only does it show an excellent synergistic effect, it also shows an excellent treatment of periodontal disease compared to existing periodontal disease prevention and treatment.

Description

Periodontal disease prevention and treatment composition

The present invention relates to a composition for preventing and treating periodontal disease.

Dental diseases are roughly divided into dental caries (cavities) and periodontal disease. Among these, periodontal disease is the most common infectious disease in adults among oral diseases, and it refers to loss of teeth due to gingival hemorrhage, swelling, periodontal sac formation, and destruction of alveolar bone. The mechanism of periodontal disease is that when the plaque is mechanically accumulated in the periodontal sac, it becomes a habitat for the bacteria present in the surrounding area, which gradually transitions from aerobic and breathable Gram-positive bacteria to anaerobic Gram-negative bacteria and proliferates deep into the periodontal sac. Will be. At this time, the toxins and all products of the proliferated anaerobic Gram-negative bacteria directly destroy tissues or stimulate the immune system and cause inflammation along with periodontal tissue destruction by various actions from the stimulated immune system.

Unlike the case of dental caries, the causative agent causing such periodontal disease is Gram-negative bacteria of the oral microorganisms, Porphymonas gingivalis, Prevotella intermedia, Actinobacillus actinomycetem. Comitans (Actinobacillus actinomycetemcomitans).

Also, the reason why these periodontal diseases appear mainly in adults is that the periodontal tissues are gradually weakened with age.

Therefore, in order to prevent or treat periodontal disease, all the causes of periodontal disease as described above can be resolved, that is, 1) restoring weakened or destroyed periodontal tissues to its original state, and 2) inflammation which is a toxic product of periodontal disease-causing bacteria. It is most preferable to use a method that can remove 3, and inhibit the causative agent of periodontal disease.

Research on the prevention and treatment of periodontal disease has been in progress since the 1920s, especially in Western countries, focusing on eliminating plaque using antimicrobial effects of antibiotics and chemotherapeutic agents (eg, chlorhexidine, cetylpyridinium). .

However, these antibiotics and chemotherapeutic agents are problematic due to side effects such as oral mucosal ulceration, exfoliative gingivitis, and pigmentation.

In addition to antibiotics and chemotherapeutic agents, enzymes that degrade plaque epilepsy and fluorides that prevent bacteria from absorbing on the tooth surface have been used for the treatment of periodontal disease, but enzymes have side effects and application methods. The formulations have a weak antimicrobial effect, which limits their universal use.

In order to solve the disadvantages of the use of synthetic drugs, including antibiotics, research has been conducted to develop drugs to treat periodontal disease using herbal extracts. As a result, Myrrha is effective for gingivitis and thrush, and Sanggenon C, an extract of the lettuce extract, inhibits the metabolism of Streptococcus mutans, resulting in the first stage of plaque formation. Although it has been found to interfere, myrrh has a weak antimicrobial effect, and the extract of Morus vulgaris has the disadvantage of inhibiting the proliferation of gingival fibroblasts to a significant level.

In addition, Sanguinaria extract has been shown to exhibit potent plaque suppression and antimicrobial effects in laboratory experiments, but clinical trial results do not provide the desired effect.

In addition, the extract of corn (Zea mays L.) is also commercially available under the trademarks such as Insadol (Dongkuk) and Dentol (Lucky), and has been used for the purpose of suppressing inflammation in the periodontal tissue.

However, the treatment of periodontal disease using the conventional herbal medicine has the advantage that there are no side effects, but it is not preferable because there is no regeneration of the periodontal tissue cells damaged by the periodontal disease, or there is no antibacterial action against the causative bacteria causing the periodontal disease. .

Therefore, the present inventors conducted intensive research using various herbal medicines to develop excellent periodontal disease treatment agents that overcome the disadvantages of the above-mentioned herbal medicines for periodontal disease treatment. In addition to excellent anti-inflammatory activity, it has been shown to exhibit strong antimicrobial activity against the causative agent of periodontal disease (Korean Patent Publication No. 94-6594), and it was also confirmed that ginkgo biloba extract showed a regenerative effect on periodontal tissue (see: Korean Patent Application No. 94-13606). The present inventors also found that the composition containing the thick leaf extract and the ginkgo leaf extract has an excellent effect on the periodontal disease in general, and has applied for a patent on the composition for treating the periodontal disease targeting them (reference: Korean Patent Publication No. 94- 14117) Subsequently, as a result of continuous research, jujube extract also showed excellent effect on the regeneration of periodontal tissue, and a patent application was also filed for the treatment of periodontal disease using jujube extract (see Korean Patent Application No. 95- 14541).

But thick extract has excellent salting and antimicrobial effect, but it has the disadvantage of lowering periodontal tissue cell activity at higher doses. Jujube extract and ginkgo biloba extract have excellent tissue regeneration effect but lack antibacterial effect, ie prevention and treatment of periodontal disease. Among the various mechanisms for, the disadvantage is that it only works through some pathways. Therefore, the present inventors searched for the prevention and treatment effect of periodontal disease by combining the jujube extract and the hindnut extract in various ratios based on the results of these studies in order to develop a treatment for periodontal disease satisfying the above requirements. The combination of these two components is superior to synergistic periodontal disease treatment effect, that is, excellent tissue regeneration, antibacterial and anti-inflammatory effect, when using each component alone in a specific combination ratio, especially thick pepper that inhibits the activity of periodontal tissue cells at high doses Overcoming the shortcomings of the extract was confirmed to exhibit an excellent tissue regeneration effect was completed the present invention.

Therefore, the present invention relates to a periodontal disease prevention and treatment composition containing the jujube extract and the thick extract as an active ingredient.

The composition of the jujube extract and the thick extract of the present invention have a wide range of effects on the prevention and treatment of periodontal disease at a specific compounding ratio, namely (1) excellent regeneration effect on weakened or destroyed periodontal tissue and (2) periodontal disease It inhibits inflammatory reactions that destroy tissues, and (3) exhibits excellent antimicrobial effect against the causative agent of periodontal disease, and their effect is much better than the effects of jujube extract and pepper extract. It provides the advantage that there is no side effect which is a problem in using.

In the composition of the present invention, as a thick gourd extract, a thick gourd extract obtained by extracting a thick gourd with an alcohol, preferably a lower alkanol such as ethanol, preferably 5.0% or more of magnolol, which is a main component, and a honokiol (honokiol) is used to contain 1.0% or more. In addition, jujube extract obtained by extracting jujube (Ziziphi fructus) in a water bath with an alcohol, preferably a lower alkanol such as ethanol, and then concentrating under reduced pressure is used.

The composition of the jujube extract and the thick gourd extract of the present invention exhibits a better synergistic treatment effect against periodontal disease than when each component is used alone. Particularly, this synergistic effect is obtained from the jujube extract and the thick gourd as demonstrated by the following experimental examples. It is most pronounced when the extract is blended in a ratio of 1: 6 to 1: 12, in particular in a ratio of 1: 8 to 1: 10.

Therefore, in the composition of the present invention, in order to maximize the synergistic treatment effect against periodontal disease, the jujube extract and the pear extract are combined in a weight ratio of 1: 6 to 1: 12, particularly preferably in a weight ratio of 1: 8 to 1: 10 use.

The composition of the present invention in which the jujube extract and the thick extract are combined in the ratio as described above may be used as it is, but may be preferably formulated into a pharmaceutical preparation by adding a conventional pharmaceutically acceptable excipient. Formulations used for this purpose can be in the form of tablets, ointments, toothpastes, solutions (gagle), sprays, and the like. For example, the composition of the present invention may be included alone in a toothpaste composition, or a pharmaceutically acceptable It can be mixed with a carrier and formulated into an internal appetite, ointment and solution to be used as a preventive and therapeutic agent for periodontal disease.

It is also included in a conventional gargle to rinse your mouth after brushing your teeth, or it can be sprayed onto a periodontal disease site as a spray containing a propellant and packed in a compressed container. You can also get a therapeutic effect.

The effective dose of the composition according to the present invention may vary depending on the purpose of use or the severity of the periodontal disease to be treated, the dosage form to be used, etc., but in general, 50 mg to 1 g per day as a combination of jujube extract and hummus extract, especially 100 Preference is given to using in amounts of from 500 mg.

The content range of the combination of the jujube extract and the thick gourd extract in the whole composition may vary depending on the purpose of use, but if the concentration is too low, the effect drops sharply, and if it is too high, the commerciality is poor due to the coloring of the jujube extract and the thick gourd extract. In consideration of the effective dose range of the combination of the jujube extract and the hind extract as described above, in the case of tablets, it is preferable to formulate to contain 50 mg to 250 mg of the combined moles of the jujube extract and the hind extract in the above ratio per unit tablet. In the case of ointments, liquids (gargles), toothpastes, and sprays, it is preferable to formulate the compounding molar in a ratio of 0.2 to 5.0% by weight.

On the other hand, jujube extract and hubac extract were LD ₅₀ (50% lethal dose) of the standard value of acute toxicity upon oral administration in mice and rats, respectively.

The composition of the present invention may further include herbal ingredients known to be effective in the treatment and prevention of periodontal disease, for example, ginkgo biloba extract, myrrh, lettuce extract, lettuce extract, corn extract and the like. You can also increase the effect.

The present invention is explained in more detail by the following examples, experimental examples and composition examples, but the present invention is not limited in any way by these.

Example 1 Preparation of Thick Pepper Extract from Thick Pepper

500 g of thin gourd (Magnoliae cortex) was thinly cut and extracted twice with reflux in a water bath using 2.5 L of 70% ethanol twice for 3 hours, the extracts were combined and filtered, and the filtrate was concentrated to give 60 g of gourd extract.

Example 2 Preparation of Jujube Extract from Jujube

300 g of Zyzyphi fructus was extracted twice with reflux in a water bath using 1.5 L of 70% ethanol, and the extracts were combined and filtered, and the filtrate was concentrated to obtain 140 g of jujube extracts.

The combination composition of the jujube extract and the thick extract of the present invention may be prepared by a conventional pharmaceutical method, for example, in the form of tablets, ointments, toothpaste liquids (gagle), sprays, and the combination ratio of the jujube extract and the thick extract is as follows. In combination with the experimental examples of preferably 1: 6 to 1: 12 by weight, particularly preferably 1: 8 to 1: 10 by weight, for example of the composition is as follows.

[Composition Example 1: Film Coated Tablet-1 Tablet (600mg)]

Jujube Extract 20mg

Thick Extract 200mg

Lactose 355mg

Hydroxypropyl Cellulose 5mg

Magnesium Stearate 3mg

Glycerin 2mg

Hydroxypropylmethylcellulose 15mg

Tablets are prepared according to the conventional method for preparing tablets using the above ingredients.

[Composition Example 2: Ointment (Value by Weight)]

Jujube Extract 0.04

Thick Extract 0.32

Carboxyvinyl Polymer 7.00

Glycerol 30.00

Peppermint Oil 0.006

Propylene Glycol 30.00

Add purified water to make 100.

The components are mixed by a conventional method for preparing an ointment to prepare an ointment.

[Composition Example 3: Toothpaste (Value by Weight)]

Jujube Extract 0.04

Thick Extract 0.40

Alumina 45.00

Propylene Glycol 10.00

Carboxymethylcellulose Sodium 2.00

Stevioside 0.30

Sodium Lauryl Sulfate 1.00

Peppermint 0.50

Add purified water to make 100.

Toothpaste is prepared by mixing the above components in a conventional toothpaste manufacturing method.

[Composition Example 4: Solution (Weight Value)]

Jujube extract 0.08

Thick Extract 0.96

Polyethylene Glycol 7.00

Ethanol 5.00

Peppermint Oil 0.06

Add purified water to make 100.

The components are mixed and dissolved in a conventional liquid preparation method, and a pH is adjusted to 5.0 to prepare a solution.

COMPOSITION EXAMPLE 5: Atomizer (Value by Weight)]

Jujube Extract 0.04

Thick Extract 0.24

Trichloromonofluoromethane 40.00

Dichlorodifluoromethane 45.00

Peppermint Oil 0.15

Ethanol 6.00

Polyethylene Glycol 8.65

The components are mixed by a conventional spray preparation method and packed into a compression container to prepare a spray.

Experimental Example: Experiment for Determination of Mixture Ratio of Jujube Extract and Thick Leaf Extract

Experimental Example 1: Measurement of gingival fibroblast activity

The effect of these formulations on the gingival fibroblast activity was varied while varying the blending ratios of the two components in the composition of the present invention containing the combination of jujube extract and the hulled extract. The gingival fibroblasts for this experiment were collected from the gingival area of the first premolar from patients who visited Seoul National University Hospital for orthodontic treatment. Immediately before the gingival area was collected, calculus and plaque were removed using a curette, washed several times with physiological saline, local anesthesia was applied to the interdental area, and normal gingival tissue was collected. The collected tissue pieces were incubated in α-MEN (GIBCO, In., Grand island, N, Y., USA) medium containing 100 U / ml penicillin, 100 µg / ml streptomycin and 100% FBS. Five passages were made while replacing the medium with the same amount of fresh medium. During the cultivation, 95% air and 5% CO ₂ were continuously supplied while maintaining a humidity of 95% and a temperature of 37C. Substituted gingival fibroblasts were treated with 0.25% trypsin-EDTA (ethylenediaminetetraacetate) solution and centrifuged at 1200 rpm to obtain cell suspension in the culture medium, and the number of cells per well in a 96-well plate using a standard hemocytometer. Inoculated to 1 × 10 ⁵ and incubated for 1 day at 5% CO ₂ , 37 ℃. The next day the culture medium was replaced with fresh culture medium, and the second day the culture medium was removed and washed with HBSS (Hank's Balanced salt solution). Platelet-induced growth factor (PDGF-BB (Genzyme, Co., Cambridge, MA., USA)), a positive control agent, was used as distilled water as a solvent, and the extract was ethanol (EtOH). Jujube extract and ginkgo leaf extract were dimethyl. After dissolving sulfoxide (DMSO) as a solvent, these were added to the culture solution so that the concentration of each test substance was as mentioned in the following table, and maintained with 95% air and 5% CO while maintaining a humidity of 95% and a temperature of 37 ° C. _The cells were incubated for 24 hours while continuing to supply _2. MTT (methyl thiazol-2-yl-2,5-diphenyltetrazolium bromide, Sigma Co., St, Louis, MO, dissolved in physiological saline after incubation was continued. USA) 50 μl of solution was added to each well and further incubated for 4 hours under the same conditions as above, and the MTT solution was filtered off under reduced pressure, and 50 μl of DMSO was added to each well to dissolve the resulting formazon crystals. Add each plate. Afterwards, the absorbance was measured at 570 nm using an ELISA reader (ELISA reader, Thermo max, molecular devices, Menlo Park CA, USA) In this experiment, only the α-MEN medium without test substance was used for each experiment. All experimental results are shown in Tables 1 to 3 calculated as a percentage of the control group.

From the results described in Table 1, the jujube extract increases the cell activity as the dose is increased, it can be seen that the maximum effect at a concentration of 10 μg / ㎖ or more.

From the results described in Table 2, it can be seen that the thick gourd extract decreases cellular activity as the dose is increased.

As can be seen from the results described in Tables 1 to 3 above, when jujube extract and thick extract are used alone, only jujube extract has the effect of increasing the activity of gingival fibroblasts and the thick extract is rather gingival fibroblast In the case of using a composition comprising a jujube extract and a thick extract extract, but the combination ratio of the weight ratio of 1: 6 to 1: 12 by weight is much better than the gingival extract when using the jujube extract alone It showed a blast activity, especially the activity of gingival fibroblasts was significantly increased at a weight ratio of 1: 8 to 1: 10 by weight. This is the result of increasing the activity of the gingival fibroblasts by the combination of jujube extract and jujube extract by using a combination of jujube extract and hulhu extract. In addition, this increased effect can be seen that far better than the effect of the composition of PDGF and ginkgo biloba extract and thick extract which are known to increase the activity of gingival fibroblasts. Therefore, it can be seen that the composition of the extract and jujube extract according to the present invention is more synergistic than the use of each component alone in a specific compounding ratio, and also clear that it is an excellent periodontal disease treatment.

Experimental Example 2: Anti-inflammatory effect measurement

The anti-inflammatory effect of the composition of the jujube extract and the thick extract according to the present invention was confirmed by measuring the inhibitory effect of PGE production of gingival fibroblasts by an in vitro test. Five to seven passages of gingival fibroblasts were placed in 1 ml of α-MEM (GIBCO, In. Grand island, N.Y., U.S.A.) medium containing 1% antibiotic, 1% FBS in each well of a 24-well plate. After dispensing at the concentration of cells, 1 ng / ml of rHUIL-1β (Genzyme, Co., Cambridge, M.A., U.S.A.) was added to induce PGE production. Here, as a test substance, the thick leaf extract was dissolved in ethanol (EtOH), the jujube extract and the ginkgo biloba extract were dissolved in dimethyl sulfoxide (DMSO) as a solvent, and the concentrations of the respective test substances were as shown in the following table. Each well was added to the culture solution, and the wells without addition of any test substance were sterilely incubated for 48 hours at 37% with 5% CO. After the incubation, the medium of each well was collected and the PGE in the medium was colorimetrically determined at 450 nm using an ELISA reader using an enzyme immunoassay system (PGEcnzymc immunoassay system, Amersham, In. Buckinghamshirc, UK). To 6 are shown.

From the results shown in Table 4, jujube extract inhibits the production of PGE as the dose is increased, it can be seen that the maximum effect at a concentration of 10 µg / ㎖ or more.

From the results described in Table 5, the thick gourd extract inhibits the production of PGE as the dose is increased, the effect can be seen that the maximum at 100 μg / ㎖ or more.

As can be seen from the results of Tables 4 to 6, the jujube extract slightly inhibits the production of PGE that mediates the inflammatory response when jujube extract and the pepper extract are used alone, and the pepper extract extracts the production of PGE. Suppresses to a significant level. However, in the case of using the composition of jujube extract and thick gourd extract showed a significantly superior PGE production inhibitory effect compared to the case of using only the thick gourd extract alone in the blending ratio of 1: 6 to 1: 12, and other combination ratio The effect was reduced at. These results are shown by increasing the PGE production inhibitory effect of the extract of jujube extract by jujube extract and jujube extract. In addition, this increased effect is far superior to the effect of the composition of the ginkgo biloba extract and thick leaf extract that is known to be the best treatment for periodontal disease. Therefore, the composition of the extract and jujube extract according to the present invention can be seen that there is a clear synergistic effect on periodontal disease compared to the case of using each component alone in a specific compounding ratio, it can be seen that it is an excellent periodontal disease treatment.

Experimental Example 3: Antibacterial Effect Measurement

Porphyromonas gingivalis (Worphyromonas gingivalis) W50, which is a donor of the periodontal pathogen, which was donated by the University of New York State University of Dentistry, to measure the antimicrobial effect of the extract of jujube extract and the pear extract according to the present invention The following experiments were performed using Prevotella intermedia ATCC 25611 and the breathable Gram-negative bacterium Actinobacillus actinomycetemcomitans Y4.

Porphyromonas Gingivalis W50 (PG) and Prebotella intermediary ATCC 25611 (PI) were inoculated into trypti-case soy agar agar medium containing 5 μl of hemin and 0.5 μl of menadione. Was incubated in an anaerobic incubator at 37 ° C., 10% CO 10% H and 80% N, and Actinobacillus actinomycetemcomitans Y4 (AA) was inoculated in trypticase soy agar agar blood medium and breathable 10 Pure culture was carried out in a bacterial incubator containing% CO. Each of the inoculated bacteria cultured for 4 to 7 days in trypticase soy broth was incubated in the anaerobic and breathable incubators as described above for 48 to 72 hours, respectively, Cells / ml. These cultured bacteria were inoculated in a blood agar medium in which each test substance was formulated at a concentration as mentioned in the following table using a multi-inoculator. These mediums were incubated at 37 ° C. in an anaerobic incubator mixed with 10% CO, 10% H and 80% N for 3 to 5 days, and then visually checked for the growth of bacteria. Minimum Inhibitory Concentration (MIC) is shown in Table 7 below.

As can be seen from the results of Table 7, the composition of the jujube extract and the pear extract, which were found to have a synergistic effect in the gingival fibroblast activity effect of Experimental Example 1 and the anti-inflammatory effect measurement Experiment of Experimental Example 2, based on the weight It was shown that the antibacterial effect was also excellent in the combination ratio of 1: 6 to 1:12 compared with the case where each was used alone.

This jujube extract itself has little antimicrobial effect, but it is the result of jujube extract by increasing the antimicrobial effect of the extract after mashing with the extract. In addition, this increased effect is better than the effect of the composition of the ginkgo biloba extract and the thick leaf extract that was previously found to be the best treatment for periodontal disease. Therefore, it is clear that the composition of the thick gourd extract and the jujube extract of the present invention have a clear synergistic antimicrobial effect compared to the case of using each component alone.

As demonstrated in the above experimental example, the composition of the jujube extract and the thick extract of the present invention has a biased effect that appears when each component is used alone at a mixing ratio of 1: 6 to 1: 12 based on the weight. Overcoming and showing various therapeutic effects on the treatment of periodontal disease, this effect was shown to be a much better synergistic effect than when using each component alone.

Experimental Example 4: Stability Test

In order to determine the stability of the formulations prepared according to Composition Examples 1 to 5, each formulation was stored under the storage conditions described below, and then the content of magnolol in the thick extract of each formulation was determined by high performance liquid chromatography (HPLC) method. Analysis at 254 nm of the UV detector and the results are shown in Table 8.

[Storage Conditions]

-Test material: The sample prepared according to Composition Examples 1 to 5 was put in a glass bottle or a compression container and sealed by sealing, then used in a cardboard box.

Storage condition: 40 ℃ (± 10 ℃)

However, the spray agent of composition example 5 was stored at room temperature.

Measurement period: Immediately after manufacture, after 2 months, after 4 months, after 6 months

-Test Items :

Composition Example 1 (Film Coated Tablet): Appearance, Disintegration (Artificial Gas Solution), Content

Composition Example 2 (Ointment): Appearance, Content

Composition Example 3 (Toothpaste): Appearance, pH (10% suspension), Content

Composition Example 4 (Solution Agent): Appearance, pH, Content

Composition Example 5 (Atomizer): Content

As can be seen from the results shown in Table 8, the stability test results show little change in the properties of tablets, ointments, toothpastes, solutions, and sprays after 6 months at 40 ° C, and the content of magnolol in the thick extract It is considered to be a stable product because it shows more than 95% compared to immediately after manufacture, and there is no problem of stability in industrialization.

Claims (7)

A composition for preventing and treating periodontal disease, comprising an alcohol extract of jujube and an alcohol extract of thick pepper in a ratio of 1: 6 to 1: 12 by weight. The periodontal disease prevention and treatment composition according to claim 1, which comprises an alcohol extract of jujube and an alcohol extract of hulk in a ratio of 1: 8 to 1: 10 by weight. The composition for preventing and treating periodontal disease according to claim 1 or 2, wherein the alcohol extract of jujube and thick pepper is an ethanol extract. The composition for preventing and treating periodontal disease according to claim 1, further comprising a pharmaceutically acceptable carrier and formulated in a pharmaceutical unit dosage form. The composition for preventing and treating periodontal disease according to claim 4, wherein the pharmaceutical unit dosage form is in the form of a tablet, ointment, toothpaste, liquid (gagle) or spray. 6. The periodontal disease prevention and treatment composition according to claim 5, comprising 50 mg to 250 mg of a combination of a refined sugar jujube extract and a thick leaf extract. The composition for preventing and treating periodontal disease according to claim 5, which comprises 0.2 to 5.0% by weight of a combination of ointment, toothpaste, solution (gagle), and spray extract of jujube extract per unit preparation.