KR101653662B1 - A pharmaceutical composition comprising extracts of sterculia lychophora for preventing, improving or treating disease of oral cavity - Google Patents

Abstract

TECHNICAL FIELD The present invention relates to a composition for preventing, ameliorating or treating oral diseases, and more particularly, to a composition containing an anti-sea extract as an active ingredient.
The anti-sea extract according to the present invention inhibits the growth of oral bacteria, the effect of inhibiting the acid production of oral bacteria, the effect of inhibiting the biofilm formation of oral bacteria, and the biofilm formation of oral bacteria on the surfaces of artificial teeth and natural teeth .

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a composition for prevention, improvement or treatment of oral diseases containing an extract as an active ingredient,

TECHNICAL FIELD The present invention relates to a composition for preventing, ameliorating or treating oral diseases, and more particularly, to a composition containing an anti-sea extract as an active ingredient.

The anti-sea extract according to the present invention inhibits the growth of oral bacteria, the effect of inhibiting the acid production of oral bacteria, the effect of inhibiting the biofilm formation of oral bacteria, and the biofilm formation of oral bacteria on the surfaces of artificial teeth and natural teeth .

The environment of the oral cavity is suitable for the growth of bacteria by nutrition and physiology. Because many bacteria are always present, the incidence of oral diseases is very high. Dental caries and periodontal disease are typical oral diseases caused by these oral bacteria.

Translucent, hard, milky white material that covers the surface of the head of a tooth and protects the tooth dentin is called tooth enamel or enamel. It is called dental caries that the tooth enamel is damaged by the acid generated by the decomposition of sugar or starch by bacteria living in the oral cavity, and tooth decay occurs.

Periodontal disease is often referred to as taste, depending on the degree of disease gingivitis (gingivitis) and periodontitis (periodontitis) is divided into. It is a relatively light and fast-acting periodontal disease called gingivitis that is limited to gingiva or soft tissue, and it is called periodontitis when the inflammation progresses to the gums and the gums.

There is also a V-shaped gap between the gum (gums) and the teeth. Periodontal disease refers to the attack of germs on the lower part of the gum line of the sulcus, damaging the periodontal ligament and adjacent tissues. As the inflammation progresses and more tissue is damaged, the groove develops into a periodontal pocket, and the more the periodontal disease becomes, the deeper the depth of the periodontal pocket becomes. As the periodontal ligament becomes deeper, periodontal ligaments become inflamed and bone loss may occur.

Typical oral bacteria include Streptococcus mutans , Porphyromonas gingivirus, gingivalis and Prevotella intermedia ). The present invention provides a composition for preventing, ameliorating or treating oral diseases through inhibition of S. mutans .

A biofilm is a layer in which microorganisms such as bacteria collect to form a single layer.

Biofilms can be made on a solid surface, or on the surface or inside of a liquid. When a microorganism forms a biofilm, the microorganism can more effectively capture and grow the nutrient, and the adhesion to the surface of the material also increases.

S. mutans It also forms a biofilm on the tooth surface. S. If mutans biofilm formation S. that becomes more and more difficult elimination of mutans, S. the activity of the mutans is enhanced. Therefore, S. The inhibition of biofilm formation of mutans can also inhibit the development of tooth decay.

Meanwhile, the objection of the present invention will be briefly described below.

In contrast, there are anti-Japanese, such as Ho Daei, Annam, and Dae Dong, and in Korea, Sterculia scaphigera Wall. In China, Sterculia lychnophora Hance: 大海 樹). In Japan, it was not found in process medicines.

There is no smell, and if you chew it for a long time, it is viscous, the taste is sweet and the quality is cold.

In addition, the opposite is done by removing the heat by lowering the lungs. Dry cough, sore throat, sore throat, osteoporosis, osteoporosis, nosebleed, stigma, ocular hypertension, toothache or hemorrhoids. Weak diarrhea and hypotensive effects were reported with pharmacological actions.

As for the prior art related to the objection of the present invention, skin whitening and antioxidant compositions containing an extract are disclosed in the Korean Patent Registration No. 10-0930592.

However, the above-mentioned document relates to a skin whitening and antioxidative use of an extract as a countermeasure, and describes a whitening cosmetic composition composed of an anti-seaweed extract and an organic solvent fraction, and a description of the efficacy of the anti- I can not.

To date, the effects of antipyretic, antipyretic, antipyretic, whitening, or antioxidant effects have been known. However, the efficacy of anti-inflammatory drugs for oral diseases has not been reported.

Accordingly, the present inventors have made the present invention to provide a composition containing an anti-sea extract as an active ingredient and having efficacy in preventing, ameliorating or treating oral diseases.

Korean Registered Patent No. 10-0930592 (December 1, 2009)

It is an object of the present invention to provide a composition for preventing, ameliorating or treating oral diseases containing an anti-sea extract as an active ingredient.

It is an object of the present invention to provide a method of inhibiting the growth of oral bacteria, an effect of inhibiting acid production of oral bacteria, an effect of inhibiting biofilm formation of oral bacteria, and an effect of inhibiting biofilm formation of oral bacteria on artificial teeth and natural teeth surfaces As an active ingredient. The present invention also provides a composition containing an anti-seaweed extract having an amino acid sequence of SEQ ID NO:

The present invention aims at solving the technical problem by providing a composition for prevention, improvement or treatment of an oral disease containing an anti-sea extract as an active ingredient.

The present invention relates to a composition for inhibiting the growth of oral bacteria, an effect for inhibiting acid production of oral bacteria, an effect for inhibiting biofilm formation of oral bacteria, and an effect for inhibiting the formation of biofilms of oral bacteria on natural teeth and artificial teeth The present invention provides a composition containing an anti-seaweed extract as an active ingredient.

The composition according to the present invention may be provided as a composition for preventing, ameliorating or treating oral diseases, and each composition may be provided as a health functional food using a foodstuff acceptable processing agent. The health functional food using the composition according to the present invention may be provided in various forms such as a functional beverage, a health supplement, tea or confectionery.

In order to use the present invention as a pharmaceutical composition, it may be prepared by a known method in the pharmaceutical field and may be mixed with itself or a pharmaceutically acceptable carrier, excipient or diluent to prepare a powder, granule, Or injections, and the like. They may also be administered orally or parenterally.

The effective dose of the composition according to the present invention can be appropriately selected depending on the degree of absorption of the active ingredient in the body, the rate of water activation and excretion, the age, sex and condition of the patient, severity of the disease to be treated,

The composition according to the present invention may be used to prepare formulations such as pills, granules, beverages, tablets or capsules, and in this case, a treating agent may be added to prepare each formulation.

The composition according to the present invention has an effect of inhibiting the growth of oral bacteria.

The composition according to the present invention has an effect of inhibiting acid production of oral bacteria.

The composition according to the present invention has an effect of inhibiting the biofilm formation of oral bacteria.

The composition according to the present invention has an effect of inhibiting biofilm formation of oral bacteria on artificial teeth and natural teeth surfaces.

Figure 1 is S. mutans And the concentration of S. mutans after treatment with the extract in concentration against the culture medium.
Figure 2 is a S. is a photograph showing a result of observing the opposed S. mutans biofilm formation pattern after treating the extract by concentration of the culture dish in a scanning electron microscope to mutans culture (bar = 10 μm. x500, Bar = 50 um 300dpi, 300dpi) .
Figure 3 is a S. mutans This is a photograph showing the formation of S. mutans biofilm after treatment with extracts at different concentrations against cultures and applied to artificial teeth.

The terms and words used in the present specification and claims should not be construed as limited to ordinary or dictionary terms and the inventor may properly define the concept of the term in order to best describe its invention It should be construed as meaning and concept consistent with the technical kill of the present invention.

Therefore, the experimental examples and the reference examples described in the present specification are merely the most preferred embodiments of the present invention and are not intended to represent all of the technical ideas of the present invention. Therefore, various equivalents and variations Examples should be understood.

Manufacturing example  1. Preparation of anti-seawater extract

The extract can be extracted with distilled water or an organic solvent, and fractionated with the above solvents. When an organic solvent is used, ethanol is preferably used, but the present invention is not limited thereto.

In the present invention, the extract was prepared by extracting the opposite sea with 70% ethanol and concentrating, and the procedure is as follows.

① In the opposite direction, ethanol was set to be 1 liter of 70% ethanol based on 300 g, and ethanol was immersed in ethanol for 2 weeks before filtration.

② The filtered material is concentrated under reduced pressure at room temperature for 15 hours to prepare an extract.

At this time, the concentrate was 10.68 g, showing a yield of 3.56%.

Experimental Example  One. S . mutans Proliferation of Inhibition  Measure

1-1. Preparation for experiment

The strains used in the experiment were Streptococcus mutans ATCC as 25175 Brain heart infusion (BHI, Difco , USA) to the liquid medium one or two primary passage after the culture inoculated to the same medium and cultured for 24 hours in a incubator of 35 ~ 40 ℃.

The strain may be replaced by other oral bacteria, and Streptococcus but is not limited to mutans .

1-2. Experimental course

Absorbance refers to the amount by which a substance absorbs light. Assuming that the intensity of the incident light to the material is I0 and the intensity of the transmitted light is I, the absorbance A is represented by A = log10 (I0 / I). When Lambert-Beer's law is established, the absorbance is proportional to the concentration of the light-absorbing material. That is, by measuring the absorbance of a substance, the concentration of the substance can be known.

On the same principle, S. When measuring the absorbance of the culture liquid containing the S mutans. The concentration of mutans , ie, S. mutans The degree of proliferation can be known. In this invention S. S In order to measure the concentration of mutans. mutans culture Absorbance was measured.

In addition, NaF was set as a positive control to increase the accuracy of the experiment.

NaF is an ionic compound which is used as a colorless solid for various purposes. Typical applications are tooth decay prevention.

The fluorine flame aids in the formation of fluoroapatite, a major component of the enamel layer of the tooth surface, and enhances the strength of the tooth. It is also added to toothpaste, and it is also sold as tablet for prevention of tooth decay.

① BHI liquid medium containing 1% glucose was prepared in group 6, and the extracts were treated at concentrations of 0.01, 0.02, 0.03 and 0.04 mg / ml respectively.

(2) One group without any treatment was set as a negative control group. The remaining one group is then set to the positive control group was treated for 0.1% NaF, S. All groups were inoculated with mutans at a concentration of 1 × 10 ⁸ CFU / mL.

③ All groups were cultured in an incubator at 35 ~ 40 ℃ for 24 hours and absorbance was measured at 550 nm using an ELISA reader (Molecular Devices Co., CF, USA) based on BHI liquid medium.

1-3. Experiment result

Figure 1 is S. mutans And the concentration of S. mutans after treatment with the extract in concentration against the culture medium.

For convenience, each experimental group was classified into 6 groups of A, B, C, D, E, (Control: A, extract: 0.01 mg / ml treated group: B, extract: 0.02 mg / ml treated group: C, extract: 0.03 mg / ml treated group: D, extract: 0.04 mg / : F)

As a result, the absorbance of the group A was the highest, and the absorbance of the group A was decreased as the concentration of the extract increased. Especially, the rate of decrease of absorbance was significantly increased from group D, and the decrease rate of group E was almost similar to that of group F, which is a positive control group.

As a result, it can be concluded that the contrary extract has an effect of inhibiting the growth of S. mutans in a concentration-dependent manner.

Experimental Example  2. S . mutans Acid production of Inhibition  Measure

2-1. Preparation for experiment

The same procedure as in Experimental Example 1-1 was carried out.

2-2. Experimental course

S. The mutans break down the sugar remaining in the tooth to form an acid, and the acid dissolves the enamel layer on the surface of the tooth to generate tooth decay. Therefore, S. By inhibiting the mutans from generating acid, it is possible to inhibit the occurrence of tooth decay. Therefore, in the present invention, pH measurement was performed to determine whether the extract of Opuntia ficus-indica inhibits acid production.

In the same conditions as Experimental Example 1-2 cultured S. The pH of the mutans was measured using a pH meter (ORIONSA 720, USA) and the inhibition of acid production was observed. In the same manner as in Experimental Example 1-2, groups without any treatment were set as negative control groups and 0.1% NaF was set as positive control groups.

2-3. Experiment result

[Table 1] shows the results of the reaction with S. mutans This table shows the pH change pattern when the culture solution is treated by concentration.

Concentration (mg / ml) pH (before incubation) pH (after incubation) Control (A) 7.24 + 0.06 5.37 ± 0.006 0.01 (B) 7.26 0.06 * 5.42 ± 0.015 * 0.02 (C) 7.25 ± 0.00 * 5.62 + 0.015 * 0.03 (D) 7.25 ± 0.00 * 6.55 0.026 * 0.04 (E) 7.25 ± 0.00 * 7.23 + 0.023 * 0.1% NaF (F) 7.30 ± 0.06 * 7.29 ± 0.000 *

For convenience, each experimental group was classified into 6 groups of A, B, C, D, E, (Control: A, extract: 0.01 mg / ml treated group: B, extract: 0.02 mg / ml treated group: C, extract: 0.03 mg / ml treated group: D, extract: 0.04 mg / : F)

Measurement results, S. No significant difference in pH was observed in all groups before the mutans were cultured. However, it was confirmed that pH was decreased in all groups after incubation, especially pH of A group was decreased to 5.37. S. mutant It can be seen that acid was generated by the culture.

In group D, the rate of decrease of pH was lower than that of A, B and C after incubation. After cultivation, the pH of group D was 6.55, which was 1.13 higher than that of group B after incubation.

After incubation, the rate of decrease of pH was very low in E group. After culturing, the pH of E group was 7.23, and it was confirmed that it was almost the same as that of F group (positive control group) after culturing.

Thus, the opposed extract was concentration-dependent with S. it is possible to conclude that there is an effect of inhibiting the acid production of mutans .

Experimental Example  3. S. mutans Biofilm formation Inhibition  Measure

3-1. Preparation for experiment

The same procedure as in Experimental Example 1-1 was carried out.

3-2. Experimental course

S. a flat dish S to observe biofilm formation patterns of mutans. Mutans were cultured and observed. The method is as follows.

① Six 35 mm dishes were prepared. Four of the dishes were treated with BHI liquid medium and the extracts were treated at concentrations of 0.01, 0.02, 0.03 and 0.04 mg / ml, respectively.

One dish without any treatment was set as negative control, and the other dish was treated with 0.1% NaF to be a positive control, and then inoculated with 5 × 10 ⁵ CFU / mL / dish of bacteria.

③ All groups were incubated at 35 ~ 40 ℃ incubator for 24 hours, and all supernatant was removed.

④ Each dish was washed with 1.5 mL of distilled water.

⑤ Fixed in 2.5% glutaraldehyde solution (in 0.1M sodium cacodylate buffer, pH 7.2, 4 ℃) for 24 hours.

⑥ Ethanol was washed at 70, 80, 95, and 100% concentration and dehydrated.

⑦ Freeze-dried, coated with gold, and observed and photographed using SEM (Scanning Electron Microscope).

3-3. Experiment result

Figure 2 is a S. mutans The culture medium opposed the treatment of extract at different concentrations is a photograph showing a result of observing the S. mutans biofilm formation pattern after the culture dish on a scanning electron microscope (bar = 10 μm. X500, Bar = 50 um 300dpi, 300dpi).

A scanning electron microscope (SEM) refers to an electron microscope having a function of directly observing a three-dimensional structure by scanning the surface of a sample with an electron beam. In the present invention, S. The mutans biofilm inhibitory effect was measured.

For convenience, each experimental group was classified into 6 groups of A, B, C, D, E, (Control: A, extract: 0.01 mg / ml treated group: B, extract: 0.02 mg / ml treated group: C, extract: 0.03 mg / ml treated group: D, extract: 0.04 mg / : F)

The results showed that the S. mutans biofilm was the thickest in group A, process The higher the concentration, the thinner the biofilm is. Especially, E group showed similar inhibition pattern of biofilm as F group, which is positive control group.

Thus, the opposed extract was concentration-dependent with S. it is possible to conclude that there is an effect of suppressing the biofilm formation of mutans .

Experimental Example  4. On the surface of artificial teeth S . mutans  Biofilm formation Inhibition  Measure

4-1. Preparation for experiment

The same procedure as in Experimental Example 1-1 was carried out.

4-2. Experimental course

In order to observe whether or not the surface of a human tooth was treated with the culture liquid, the same results as those obtained when observation of the dish culture were observed, artificial teeth made of the same constituents and composition as human teeth were prepared. The artificial teeth S. In contrast to the mutans culture, the extracts were treated by concentration and applied.

1) The six artificial teeth (Endura, Shofu Inc., Kyoto, Japan) were prepared. Four of them were inoculated with BHI liquid medium containing 0.1% sucrose and the extracts were added at concentrations of 1, 5, 10 and 50 ug / mL Respectively.

2) Set the one artificial teeth did not have any treatment as a negative control, and then the remaining one artificial teeth is set as a positive control by applying a ^{0.1% NaF, 5 × 10 5} CFU / mL S such that the number of bacteria in / dish. mutans Respectively.

3) All groups were cultured in incubator at 35 ~ 40 ℃ for 24 hours. All the supernatant was removed.

4) Dissect each artificial tooth with 1.5 mL of 0.1% Safranine solution for 30 seconds.

5) After removing the 0.1% Safranine solution, it was washed twice with 1.5 mL of distilled water and dried.

4-3. Experiment result

Figure 3 is a S. mutans This is a photograph showing the biofilm formation pattern of S. mutans after treatment with extracts in concentration against the culture solution and applied to the artificial teeth.

The results showed that S. mutans biofilm was the most abundant in group A, and that in group B, process As the concentration increases, the formation rate of S. mutans biofilm is remarkably decreased. Especially, E group showed similar inhibition pattern of biofilm as F group, which is positive control group.

As a result, the contrary extracts were found to be S. and the effect of inhibiting biofilm formation of mutans, S in natural tooth surfaces. It can be concluded that mutans are also effective in inhibiting the formation of biofilms.

Claims (5)

A composition containing an anti-body extract as an active ingredient,
The composition comprises an anti-sea extract having a concentration of 0.01 to 0.04 mg / ml,
The effect of suppressing the proliferation of Streptococcus mutans, the inhibitory effect of Streptococcus mutans on the production of acid, the inhibitory effect of Streptococcus mutans on the biofilm formation, and the effect of Streptococcus mutans A pharmaceutical composition for preventing, ameliorating or treating oral diseases caused by Streptococcus mutans by having an effect of inhibiting biofilm formation.
delete delete The method according to claim 1,
The pharmaceutical composition for preventing, ameliorating or treating oral diseases caused by Streptococcus mutans, wherein the anti-seaweed extract is an ethanol extract of the opposite side.
A composition containing an anti-body extract as an active ingredient,
The composition comprises an anti-sea extract having a concentration of 0.01 to 0.04 mg / ml,
The effect of suppressing the proliferation of Streptococcus mutans, the inhibitory effect of Streptococcus mutans on the production of acid, the inhibitory effect of Streptococcus mutans on the biofilm formation, and the effect of Streptococcus mutans A health functional food composition for improving oral disease caused by Streptococcus mutans by having an effect of inhibiting biofilm formation.

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