KR20190127447A - A composition for preventing and treating oral disease comprising black rice - Google Patents

Abstract

The present invention can be utilized as a pharmaceutical composition or health functional food by effectively acting on the inflammation inhibitory activity of Aggregatibacter actinomycetemcomitans, pathogen of oral disease, using a black rice extract as an active component.

Description

A composition for preventing and treating oral disease comprising black rice}

The present invention relates to a composition for the prevention and treatment of oral diseases including periodontitis. More specifically, the black rice is an effective ingredient for the prevention and treatment of oral diseases with inflammatory inhibitory activity against Aggregatibacter actinomycetemcomitans , the causative agent of oral diseases. It relates to a composition for.

Oral diseases, which are a worldwide problem with oral health, are dental caries and periodontitis. WHO reports that dental caries and periodontal disease occur in about 60% of the adult population worldwide (Petersen, 2003, 2004).

The primary cause of oral disease is known as bacteria present in the dental plaque (Marsh, 1992). Dental caries is caused by demineralization of tooth enamel by organic acids such as lactic acid, a sugar metabolite of bacteria present in the dental plaque . Mutans group streptococci (mutans streptococci) have been reported (Loesche, 1986). Periodontitis is known as Gram-negative species such as Porphyromonas gingivalis (P. gingivalis) , Prevotella intermedia (P. intermedia) , Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans), and Fusobacterium nucleatum (F. nucleatum). .

Compounds widely used to prevent oral disease while inhibiting the growth of oral bacteria include chlorhexidine, fluorine compounds, xylitol and sorbitol. Chlorhexidine is an antibiotic used in periodontal treatment to inhibit gram-negative bacteria. As a side effect, chlorhexidine causes pigmentation in teeth and prostheses, and causes taste disorders and burning in the oral mucosa (Tredwin et al . 2005). Fluoride compounds are most commonly used to inhibit dental caries-causing bacteria, but excessive intake can cause abdominal pain, spots and cancer (Spencer and Do, 2008).

Therefore, recently, in order to solve the above problems, an oral composition having excellent pharmacological action and stability has been developed in addition to the conventional oral cleaning action.

Registration No. 10-1641213 relates to an oral composition for preventing and treating periodontitis, including cedar extract and arrow extract as active ingredients, showing growth inhibition activity of P. gingivalis, the cause of periodontitis. 10-2011-103267 relates to an oral composition for the prevention and treatment of caries or periodontitis using turmeric leaf extract as an active ingredient and exhibits antimicrobial activity against S. mutans .

Black rice or black medicinal rice is processed into various forms of food due to its unique color and aroma, and consumption is also increasing. Black rice contains antioxidants such as anthocyanins, polyphenols, flavonoids, and vitamins. Ethanol extracts of black rice have been reported to have superior antioxidant effects than white rice (Choi SW et al. 1996. Food Biotechnol 5: 305-309, Nam SH et al . 1997. Agri Chem. Biotechnol 40: 307-312). On the other hand, the study showed that the administration of black rice bran pigment to the rats made with rice germ improves lipid metabolism and increases the antioxidant enzyme activity (Cho Min-Kyung et al., 2008. J Korean Soc.Appl Biol Chem 51 (3): 200 -206), black rice extract and anthocyanin have been reported to lower blood and liver cholesterol levels in rats fed a high cholesterol diet.

The black rice having the above-mentioned efficacy has been mainly applied to the health functional food mainly in the past, and there are some examples of applying the black rice extract as a cosmetic composition in Korean Patent Publication No. 10-2005-0075516, but preventing and treating oral diseases including periodontitis. There is no disclosure or teaching about the effects and methods for preparing the composition therefor.

Patent Registration No. 10-1641213, Publication No. 10-2011-103267, Publication No. 10-2005-0075516

Min-Kyung Cho et al., 2008. J Korean Soc. Appl Biol Chem 51 (3): 200-206 Choi SW et al. 1996. Food Biotechnol 5: 305-309, Nam SH et al. 1997. Agri Chem. Biotechnol 40: 307-312

The present invention is to solve the problem to provide a composition, health functional food or pharmaceutical composition that is effective in the treatment and prevention of oral diseases, including periodontitis, using black rice extract as an active ingredient.

In order to solve the above problems, the present invention provides a composition for preventing and treating oral diseases, pharmaceutical compositions, and health functional foods using black rice extract as an active ingredient as a means for solving the problems.

The pharmaceutical composition of the present invention, the health functional food is characterized in that it comprises the step of extracting black rice bran with ethanol, filtering and concentrating with a vacuum rotary concentrator and dissolving in (dimethylsulfoxide, DMSO) in dimethyl sulfoxide.

Black rice extract according to the present invention is not only very high inhibitory activity against the periodontitis-inducing bacteria A. actinomycetemcomitans , but also can be used in various compositions such as oral disease prevention and treatment, pharmaceutical compositions and health functional foods.

1 is a diagram showing the results of black rice extract inhibits the inflammatory mediator (NO) induced by the periodontitis causing bacteria (A. actinomycetem comitans) .
2 to 4 is a view showing the inhibitory effect of inflammatory cytokines IL-1β, IL-6, TNF-α expression by the periodontitis causing bacteria of the black rice extract.

Hereinafter, the present invention will be described in more detail with reference to the accompanying drawings and examples.

Although the present invention may be modified in various ways and may have various embodiments, the present invention is not intended to be limited to the specific embodiments, and includes all changes, equivalents, and substitutes included in the spirit and technical scope of the present invention. It should be understood to do. In the following description of the present invention, if it is determined that the detailed description of the related known technology may obscure the gist of the present invention, the detailed description thereof will be omitted.

Terms such as first and second may be used to describe various components, but the components should not be limited by the terms. The terms are used only for the purpose of distinguishing one component from another. For example, without departing from the scope of the present invention, the first component may be referred to as the second component, and similarly, the second component may also be referred to as the first component. The term and / or includes a combination of a plurality of related items or any item of a plurality of related items.

 The terms used in the present application are not intended to limit the present invention. Singular expressions include plural expressions unless the context clearly indicates otherwise. In this application, the terms "comprise" or "have" are intended to indicate that there is a material, effect, potency, mechanism of action, or a combination thereof, as described in the specification, including one or more materials, effects, potency, It should be understood that it does not exclude in advance the possibility of the presence or the addition of a mechanism of action or combinations thereof.

Unless defined otherwise, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art. Terms such as those defined in the commonly used dictionaries should be construed as having meanings consistent with the meanings in the context of the related art, and shall not be construed in ideal or excessively formal meanings unless expressly defined in this application. Do not.

Example 1: Preparation of Black Rice Extract

100 g of washed and dried black rice bran was extracted using 1000 ml of 95% ethanol as the extraction solvent. The extracted sample was filtered through No. 2 Whatman filter paper, and the filtered extract was concentrated with a vacuum rotary concentrator to remove the solvent component, thereby preparing a black rice extract.

Experimental Example 1: Periodontitis causing bacteria of black rice extract A. actinomyecetem comitans Inhibition of inflammation by

To measure the anti-inflammatory activity of the black rice extract, macrophages (RAW264.7) were dispensed in 24-well plates at 5 x 10 ⁵ cells / ml per well and incubated in a 37 ° C, 5% CO₂ incubator for 24 hours. Then, after pretreatment with 0, 0.0625, 0.125, 0.25, 1, 2 mg / ml black rice extract prepared in Example 1, 0.5 × formal saline treated Aggregatibacter actinomyecetemcomitans ( A. actinomycetemcomitans) 1 × 10 ⁸ cfu Then, incubated for 24 hours. After incubation for 24 hours, 100 μl of supernatant was dispensed into a 96well plate, reacted with a Griess reagent to measure nitric oxide (NO) concentration, and absorbance at 540 nm was measured using an ELISA reader. The concentration was obtained.

A. actinomycetem comitans (A. a.) NO inhibition was calculated as follows.

×100 % NO inhibition rate = 1-

× 100%

¹⁾ A. a. : A. actinomycetemcomitans

The results are shown in Table 1 and FIG. As a result, NO concentration was 8.22 ± 0.00 μM in the control group, 8.12 ± 0.00 μM in the 0.0625 mg / ml black rice extract alone group, 7.56 ± 0.00 μM in the 0.125 mg / ml, 8.03 ± 0.00 μM in the 0.25 mg / ml, 0.5 mg / ml 7.51 ± 0.00 μM in ml, 8.12 ± 0.00 μM in 1 mg / ml, 7.99 ± 0.00 μM in 2 mg / ml, and the NO concentration rose to 18.53 ± 0.00 μM in the A. actinomycetemcomitans alone group and A. actinomycetemcomitans + black rice Extract 17.68 ± 0.00 μM at 0.0625 mg / ml, 18.01 ± 0.00 μM at 0.125 mg / ml, 18.48 ± 0.00 μM at 0.25 mg / ml, 16.92 ± 0.00 μM at 0.5 mg / ml, 13.42 ± 0.00 μM at 1 mg / ml , 2 mg / ml showed 9.45 ± 0.00 μM.

Black Rice Extract Inhibits Inflammatory Mediator (NO) Induced by Periodontitis sample Nitrile oxide (μM) % Inhibition  Negative Control  8.22 ± 0.00 - One  Black Rice Extract 0.0625 (mg / ml)  8.12 ± 0.00 - 2  Black Rice Extract 0.125 (mg / ml)  7.56 ± 0.00 - 3  Black Rice Extract 0.25 (mg / ml)  8.03 ± 0.00 - 4  Black Rice Extract 0.5 (mg / ml)  7.51 ± 0.00 - 5  Black Rice Extract 1 (mg / ml)  8.12 ± 0.00 - 6  Black Rice Extract 2 (mg / ml)  7.99 ± 0.00 - 7 A. a. ¹⁾ single administration group 18.53 ± 0.00 - 8  A. a. + Black Rice Extract 0.0625 (mg / ml) 17.68 ± 0.00 * 8.25% 9  A. a. + 0.125 mg / ml black rice extract 18.01 ± 0.00 * 5.04% 10  A. a. + Black rice extract 0.25 (mg / ml) 18.48 ± 0.00 * 0.45% 11  A. a. + Black Rice Extract 0.5 (mg / ml) 16.92 ± 0.00 * 15.59% 12  A. a. + Black Rice Extract 1 (mg / ml) 13.42 ± 0.00 * 49.54% 13  A. a. + Black Rice Extract 2 (mg / ml)  9.45 ± 0.00 * 88.07% ¹⁾ A. a. : A. actinomycetemcomitans

Experimental Example 2: Method and result of measuring the effect of black rice extract on the cytokine expression

In order to measure anti-inflammatory cytokines of black rice extract, macrophages (RAW264.7) were dispensed in a 60m dish at 2 × 10 ⁵ cells / ml for each well and incubated in a 37 ° C., 5% CO₂ incubator for 24 hours. After pretreatment with 0, 1, 2 mg / ml of black rice extract prepared in Example 1, 0.5% formal saline-treated periodontal causative agent A. actinomycetemcomitans 1 × 10 ⁸ cfu was incubated for 24 hours. After trizol treatment, RNA was extracted and the expression of IL-1β, IL-6, and TNF-α (see Table 2) was measured by real-time PCR. The results are shown in Table 3 and FIG.

As a result, IL-1β showed increased expression of 90.39 ± 8.45, IL-6897.15 ± 75.38, and TNF-α 5.92 ± 0.55 when only periodontal causative organisms were treated, but black rice extract was administered at 2mg / ml concentration. When IL-1β induced by periodontal causative bacteria was 2.25 ± 0.56, IL-6 0.86 ± 0.86, TNF-α 0.34 ± 0.08 was confirmed that the expression is suppressed.

 Primer Sequences of inflammatory genes Gene Primer sequence IL-1β Forward 5′-GAAAGACGGCACACCCACCCT-3 ′ Reverse 5′-GCTCTGCTTGTGAGGTGCTGATGTA-3 ′ IL-6 Forward 5′-TCAGAATTGCCATTGCACA-3 ′ Reverse 5′-GTCGGAGGCTTAATTACACATG-3 ′ TNF-α Forward 5′-GGCAGGTCTACTTTGGAGTCATTGC-3 ′ Reverse 5′-ACATTCGAGGCTCCAGTGAATTCGG-3 ′ β-actin Forward 5′-CATCACTATCGGCAATGAGC-3 ′ Reverse 5′-GACAGCACTGTGTTGGCATA-3 ′

Black Rice Extract Inhibits the Expression of Inflammatory Cytokines IL-1β, IL-6, and TNF-α Induced by Periodontitis Real-Time PCR (gene expression folds of control) Sample
Primer Cytokines Control A.a. ^One) Alone A.a. + black rice (2 mg / ml) IL-β 1 ± 0.55 90.39 ± 8.45 2.25 ± 0.56 * IL-6 1 ± 0.69 897.15 ± 75.38 0.86 ± 0.86 * TNF-α 1 ± 0.61 5.92 ± 0.55 0.34 ± 0.08 *

¹⁾ A. a. : A. actinomycetemcomitans

Through the above experimental example, it can be seen that the black rice extract of the present invention is very effective in preventing and treating oral disease including periodontitis by inhibiting the expression of A. actinomyecetemcomitans , which is the cause of periodontitis.

The black rice extract can be prepared by oral composition or functional health food. More specifically, oral compositions can be prepared with oral detergents containing sodium fluoride, ethanol, glycerin, citric acid and peppermint, sorbitol solution, glycerin, propylene glycol, fatty acid esters, sodium sulfate, sodium chloride, flavorings, carboxymethylcellulose It may be prepared as a toothpaste containing sodium, and may be prepared as chewing gum by containing sugar, glucose, starch, glycerin, and flavorings.

Functional health foods can be prepared by mixing the above black rice extract with vitamins, biotin, nicotinic acid amide, folic acid, calcium, mineral mixtures, zinc oxide, potassium, sodium citrate, magnesium chloride.

While the invention has been described with reference to exemplary embodiments so far, those skilled in the art will appreciate that various changes can be made therein and equivalents may be substituted for elements thereof without departing from the scope of the invention. You will know.

In addition, many modifications may be made to adapt a particular situation and material to the teachings of the invention without departing from the essential scope thereof. Therefore, it is intended that the present invention not be limited to the particular embodiments disclosed as the best mode contemplated for carrying out the invention, but that the invention will include all embodiments falling within the scope of the appended claims.

Black rice extract of the present invention can be utilized in various ways, such as pharmaceutical compositions and health functional food for dental health.

Claims (3)

Composition for the prevention and treatment of oral diseases with black rice extract as an active ingredient.
The method of claim 1,
The oral disease is a composition for the prevention and treatment of oral diseases, characterized in that the causing bacteria Aggregatibacter actinomycetemcomitans .
Extracting the black rice bran with ethanol;
Filtering the extracted black rice bran and concentrating with a vacuum rotary concentrator; And
After removing the solvent component of the concentrated solution, the method for producing a composition for the prevention and treatment of oral diseases comprising the step of dissolving in dimethyl sulfoxide (DMSO).

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