KR101544939B1 - Composition for Prevention or Treatment of Dental Caries Comprising Extract of Gentianae macrophyllae Radix - Google Patents

Abstract

TECHNICAL FIELD The present invention relates to a composition for preventing or treating dental caries, and more particularly, to a composition for oral care and a health food for preventing or treating dental caries comprising an extract of Chrysanthemum morifolium as an active ingredient. The above extract of Jinhyo has excellent antimicrobial effect against the teeth of the teeth and inhibits the activity of glucosyltransferase which is an enzyme released during the growth and growth of the teeth of the teeth, So that it can be effectively used as an oral composition or a health food for preventing and / or treating dental caries.

Description

TECHNICAL FIELD The present invention relates to a composition for preventing or treating dental caries,

TECHNICAL FIELD The present invention relates to a composition for preventing or treating dental caries, and more particularly, to a composition containing an extract of Dental caries as an active ingredient and useful for preventing and / or treating dental caries.

Dental caries is a disease in which the hard tissues of teeth are destroyed by the action of locally produced organic acids due to the metabolism of carbohydrates in foods due to microorganisms present in the dental plaque of dentate gingiva and oral hygiene. It is one of the world's worries as well as our country. Dental caries is not directly related to human life, but it affects the digestive system diseases directly or indirectly because the chewing function of the food is deteriorated. The prevalence of dental caries is particularly high among children, with most people experiencing or experiencing pain from dental caries, to as high as 85% of 5-year-olds.

The cause of dental caries is various but generally lactic acid which is formed by decomposition of carbohydrate such as sugar or starch of food waste attached to teeth due to the fermentation action of bacteria resident in oral cavity causes tooth decay of hard tissue lime, . Several pathogenic microorganisms causing such dental caries are known, but in particular Streptococcus mutans mutans ), Streptococcus sobrinus sobrinus , etc. are the main pathogen of dental caries development (Non-Patent Document 1). Streptococcus mutans secretes an enzyme called glucosyltransferase (GTase) in the outer layer of mycelia or in the surface of mycelia to produce glucose and fructose using sucrose as a substrate in the food, and at the same time, insoluble glucan Is formed on the tooth surface. This glucan attaches to other microorganisms in the oral cavity to form a dental plaque, ie, a dental plaque. In this region, lactobacillus including Streptococcus mutans is proliferated to produce lactic acid from fructose as a carbon source, and lactic acid is captured and concentrated in adherent glucan. So a high concentration of a concentrated acid of the calcium phosphate composition of the tooth melts the Ca ^{2 +} unloaded by causing it to be a dental caries sikimeuroseo dissolving the outer membrane of the enamel of the teeth (Non-Patent Document 2).

Methods for suppressing such dental caries include using antibiotics such as sporamycin, vancomycin and chlorhexidine, inhibiting pathogenic bacteria by organic or inorganic fluorine (Non-Patent Document 3). However, when the antibiotics are continuously used, there is a disadvantage that the microorganisms may become resistant, or side effects such as diarrhea and vomiting may be caused. In addition, there is a problem in that the abuse of such antibiotics inhibits the growth of the bacteria in the oral cavity have. Therefore, there is an urgent need to develop an effective and stable dental caries prevention and treatment agent.

On the other hand, Gentianae macrophyllae Radix refers to the root of perennial herbaceous plants of buttercups and related plants that grow in submerged soil in the bushes of Korea. It is also called "Jinmu" or "Ookdogi". It is a non-toxic herbal medicine used as a medicine that is picked in fall and dried in the sun by removing foliage and whisker root. In the oriental medicine, it has been known from time to time that the above-mentioned in-situ relatives can be used for the treatment of analgesia, jinbok, customary vinegar, arthritis (Patent Document 1), 黄 疽 and urine disadvantages In recent years, it has been known that besides the above-mentioned pharmacological effects, it has a blood pressure lowering effect, antibacterial action against various infectious diseases, and inflammation due to increase of analgesic and adrenocorticotropic hormone secretion (Patent Document 2). In addition, it is known that the extract of Jincyung can be used for the treatment of inflammatory diseases (non-patent document 4) or for the treatment of irritable skin diseases, although it has been known that it is used as a component of acne or anti- (Patent Document 4), there has been no report showing that it can be used for prevention or treatment of dental caries.

The inventors of the present invention have studied various herbal medicine extracts which can be useful for the prevention and treatment of dental caries. As a result, it has been found out that the extract of Jinhyo has excellent dental caries prevention or therapeutic effect, Thereby completing the invention.

Korean Patent Publication No. 10-2003-0063106 Korean Patent Publication No. 10-2000-0043996 Korean Patent Laid-Open No. 10-2003-0044505 Korean Patent Publication No. 10-2006-0102620

 Liljemark W.F. et al., Crit. Rev. Oral Bio. Med. 7: 180-198, 1996  Sigmund, S.S. et al., J. Periodontol. 63: 322-331, 1992  McBain, A.J. et al., Appl. Environ. Microbiol. 69: 4770-4776, 2003  Cho, Hee-Chang et al., Korean Journal of Dermatology, Vol. 22, No. 3, 63-70, 2009

It is intended to provide a composition for the prevention or treatment of dental caries, which contains an extract of Chiono suppressant as an active ingredient.

In order to accomplish the above object, the present invention provides an oral composition for preventing or treating dental caries comprising an extract of Chrysanthemum morifolium as an active ingredient.

The present invention also provides a health food for preventing or alleviating dental caries comprising an extract of Chrysanthemum chrysanthemum as an active ingredient.

The present invention provides a composition for oral use for preventing or treating dental caries comprising an extract of Chamaecyparis obtusa as an active ingredient.

It is preferable that the above-mentioned crude extract is extracted by adding 2 to 30 times, preferably 5 to 20 times, of the extraction solvent to the true extract. As the extraction solvent, water is generally used, but other organic solvents than water may be used as needed. Examples of the organic solvent that can be used include C ₁ to C ₄ lower alcohols, preferably ethanol, acetone, or an aqueous solution thereof, but the present invention is not limited thereto. In the present invention, the above-mentioned "extract of Jinjiao " is used not only to extract the Jinjae extract prepared by using the above-mentioned extractant but also to include fractions obtained by further adding one or more" fractionated organic solvent " The fractionated organic solvent is not particularly limited, and conventional organic solvents can be used without limitation. In a preferred embodiment of the present invention, an organic solvent such as ethyl acetate or butanol is used as a fractionated organic solvent for the water extract of Jin-Ryang.

The oral composition containing the extract of the present invention can be used as a preparation for oral administration, such as dentifrices, mouthwashes, or mouthwashes, but is not limited thereto. In the oral composition of the present invention, the dentifrice may contain one or more components such as an abrasive, a binder, a moisturizer, a foaming agent, a sweetener, and a flavoring agent. Examples of the abrasive include aluminum hydroxide, silicic anhydride, aluminum silicate, calcium secondary phosphate, secondary hydroxide and anhydride, calcium tertiary phosphate, calcium carbonate, calcium pyrophosphate, insoluble sodium metaphosphate, magnesium tertiary phosphate, magnesium carbonate, Calcium, polymethyl methacrylate, and other synthetic resins may be used in an amount of about 20 to 90% of the total composition (20 to 60% in the case of paste toothpaste). The above-mentioned viscous solution may be prepared by a method including, for example, stirring of carrageenan, various incremental cellulose derivatives, xanthan gum and tragacanth gum, synthesis of polyvinyl alcohol, sodium polyacrylate, polyacrylic acid / maleic acid copolymer, carboxyvinyl polymer Polymer derivatives, and organic binder such as silica and laponite, in an amount of about 0.3 to 5% of the total composition. In the paste and liquid oral compositions of dentifrices, one or more of sorbitol, glycerin, ethylene glycol, propylene glycol, polyether glycol, polypropylene glycol and the like may be used as a moisturizing agent in the preparation of the composition in the range of about 10 to 70% And the like.

On the other hand, the oral composition containing the crude extract of the present invention may contain at least one selected from the group consisting of Mentol, Anetol, Eugenol, Limonene, Ocimene, n-dodecyl alcohol (n -Dodecy Alcohol, Citronenol, alpha-Terpineol, Methyl salycilate, Methyl Acetate, Citronelyl Acetate, Cineol, Linalool, Ethyl Linalool, Vanillin, Thymol, Speamint Oil, Sage Oil, Rosemary Oil, Cinnamon Oil, Or the like may be used alone or in combination in an amount of 0.1 to 10%, preferably 0.5 to 5%, of the whole composition. In addition, sweeteners such as sodium saccharin, stevioside, aspartame, D-xylose and the like may be used in an amount of 0 to 1%, preferably 0.01 to 0.5%, of the total composition.

The oral composition of the present invention may also contain a cationic fungicide such as triclosan, chlorohexidine, benzethonium chloride, benzalkonium chloride, cetylpyridium chloride, alkali metal monofluoro such as sodium monofluorophosphate and potassium monofluorophosphate, Phosphate, fluoride such as sodium fluoride and tin fluoride, tranexamic acid, epsilon aminocaproic acid, aluminum chlorohydrosilate allantoin, dihydrocholestanol, clitoridic acid, glycerophosphate, sodium chloride, water soluble inorganic phosphate compound , Ascorbic acid (sodium), dl-a-tocopherol acetate, xylitol, pyridoxine hydrochloride, polyvinylpyrrolidone, and the like.

In addition, anionic surfactant, nonionic surfactant, and / or cationic surfactant may be used as a surfactant used as a foaming component in the oral composition of the present invention. Examples of the anionic surfactant include sodium alkyl sulfate such as sodium lauryl sulfate and sodium myristyl sulfate, sodium N-lauroyl sarcosinate, sodium N-miristyl sarcosinate, Sodium N-acyl sarcosinate and the like can be used. Nonionic surfactants include saccharide-derived fatty acid esters such as sugar fatty acid esters, maltose fatty acid esters and lactose fatty acid esters, poly Oxyethylene hardened castor oil, polyoxyethylene higher alcohol ether, polyoxyethylene polyoxypropylene copolymer, polyoxyethylene polypropylene fatty acid ester, and the like. Examples of the cationic surfactant include N-alkyldiaminoethylglycine such as N-lauryldiaminoethylglycine, N-myristyldiaminoethylglycine and the like, N-alkyl-N-carboxymethylammonium betaine, -Hydroxyethylimidazoline betaine sodium and the like can be used. In this case, anionic surfactant is mainly used as the foaming component, and sodium alkylsulfate such as sodium lauryl sulfate is the most widely used. These surfactants are preferably used singly or in combination of two or more, and the amount thereof is usually 0.01 to 0.5%, more preferably 0.05 to 0.3%, of the total composition.

The extract may be contained in an amount of 0.01 to 50% by weight based on the total weight of the oral composition of the present invention, but its content may be appropriately adjusted depending on the degree of absorption of the active ingredient, age, sex, condition, .

According to one embodiment of the present invention, the extract of Chinchilla japonica exhibits strong antimicrobial activity against causative bacteria of dental caries, and exhibits strong antitumor activity in the ethyl acetate fraction and butanol fraction of Chinatown water extract (see Table 1). In addition, in the antimicrobial activity experiment using the disk diffusion method, a visible inhibitory ring was observed in the 70% ethanol extract, and it was confirmed that the growth of the teeth was strongly inhibited (see FIG. 1).

In addition, according to another embodiment of the present invention, glucan production was most strongly inhibited in the hydrothermal extract-treated group, which had weak antibacterial activity against the Toxin bacteria, and the glucan production inhibition was relatively strong even in the ethyl acetate fraction, (See Table 2), the ethyl acetate fraction exhibited a similar inhibitory activity to the hot-water extract even at a dilution of 2-fold.

In addition, according to another embodiment of the present invention, the phenolic compound content was the highest in the ethyl acetate fraction, similar to the antimicrobial activity level confirmed in the treatment of the Toxoplasma gondii, and a large amount of the phenolic compound was also present in the butanol fraction , Suggesting that the antimicrobial activity of chinensis against the tooth whiteness may be due to the phenolic compounds present in the syngas (see Table 3).

In addition, according to another embodiment of the present invention, as a result of toxicity test using a mouse, no abnormal findings of clinical symptoms related to the administration of the test substance were observed in all of the mice used in the experiment, and no dead animal was observed Reference). together. No significant weight loss or increase effects were observed in the experimental animals (see Table 5).

The present invention also provides a health food for preventing or alleviating dental caries comprising an extract of Chrysanthemum chrysanthemum as an active ingredient.

The extract can be added to a health food for the purpose of preventing or improving dental caries symptoms. When the extract is used as a food additive, it can be added as it is or can be used in combination with other food or food ingredients. Can be appropriately used. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health or therapeutic treatment). In general, when the food or beverage is produced, the crude extract is added in an amount of not more than 30% by weight, preferably not more than 10% by weight based on the raw material. However, in the case of long-term consumption intended for health and hygiene purposes or for health control purposes, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range .

There is no particular limitation on the kind of the food. Examples of the food to which the above substance can be added include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, ice cream, various soups, drinks, tea, Alcoholic beverages, and vitamin complexes, all of which include health foods in a conventional sense.

The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. The above-mentioned natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like. The ratio of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 ml of the crude extract.

In addition to the above, the health beverage composition of the present invention may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, Alcohols, carbonating agents used in carbonated drinks, and the like. In addition, the extract may contain flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. Although the ratio of such additives is not critical, it is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the extract.

As described above, the extract of the present invention has an excellent antimicrobial effect against tooth whitening bacteria, suppresses the activity of glucosyltransferase, which is an enzyme released during the growth and growth of bacteria, and its effect is continuous But it is extracted from a natural product and is very safe. Therefore, it can be effectively used as an oral composition or a health food for preventing and / or treating dental caries.

FIG. 1 (A) shows that the growth inhibition ring of 22 mm was formed when 2 mg of 70% ethanol extract was formed, and (B) the growth inhibitory effect of the extract of Angelica keiskei koidz Indicates that a growth inhibition ring of 13 ㎜ was formed when 2 mg of hot - water extract was administered.

Hereinafter, the present invention will be described in detail with reference to examples.

However, the following examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

Example  One: Whole life  Preparation of extract

<1-1> Heat number  Preparation of extract

One liter of distilled water was added to 50 g of dried persimmon, and the mixture was subjected to hot-water extraction for 3 hours, followed by filtration using a filter paper and lyophilization.

<1-2> Preparation of organic solvent extract

After adding 0.5 liter of 70% ethanol to 50 g of dried persimmon, it was sonicated at 25 ° C for 1 hour, filtered using a filter paper, and the solvent was removed using a rotary evaporator under reduced pressure.

<1-3> Heat number  Active ingredient of extract Fraction  Produce

The hot water extract prepared in Example <1-1> was suspended in water and fractionated with a solvent having a low polarity and a solvent having a high polarity using ethyl acetate and butanol in consonance with the polarity of the solvent, Organic solvent fractions were obtained. The obtained organic solvent fractions were concentrated using a rotary evaporator under reduced pressure and dissolved in DMSO (Dimethyl Sulfoxide) at a concentration of 25 mg / ml and stored at -20 ° C.

Comparative Example  One. goldthread  Preparation of extract

In the same manner as in Example <1-1>, except for using antimicrobial activity of antimicrobial activity against chrysanthemum strain instead of antimicrobial antimicrobial activity, the extract of Antimony nucifera was prepared.

Experimental Example  1. Measurement of growth inhibitory activity against causative bacteria of dental caries

Streptococcus mutans, a causative organism of dental caries used in this experiment, is a gram-positive, tuberous anaerobic bacterium. KCTC3065 strain, which has been distributed from the Center for Microbial Resources, Korea Research Institute of Bioscience and Biotechnology, was purchased from BHI Broth (Difco, USA) The cells were subcultured on Agar medium and used for experiments. In order to investigate the growth inhibitory activity of the extracts against dental caries, the Streptococcus mutans strain was inoculated on BHI broth and subcultured, and then the bacterial solution was diluted so that the absorbance value at 550 nm was 0.1. This dilution was inoculated 10% in BHI broth and 200 [mu] l in 96-well microplates. After the dispensation, the final concentration was 1.25 mg / ml for the 70% ethanol extract and hot water extract, and 0.625 mg / ml for the hot water extract-derived organic solvent fraction, respectively. After incubation for 20 hours, the absorbance was measured and the growth inhibition was measured. The results are shown in Table 1.

Treatment concentration
(Mg / ml) 70% ethanol
extract Hot water extract Ethyl acetate
Fraction Butanol fraction 1.25 1.25 0.625 0.625 Tooth root
Growth Inhibition (%) 57.30 ± 0.73 11.78 ± 1.34 53.92 + - 2.23 56.34 ± 0.86

As can be seen from Table 1, the strongest antimicrobial activity was confirmed in the 70% ethanol extract. In addition, when the ethyl acetate fraction and butonol fraction of the water extract were treated, the anti-periodic bacterial activity was confirmed to be about 5 times stronger than that of the hot water extract of Shin-kyo as the mother liquor, And the efficiency was improved.

Experimental Example  2. Measurement of antimicrobial activity against causative bacteria of dental caries

The antibacterial activity of the crude extract prepared in Example 1 was measured by the disk diffusion method. To this end, the BHI plates supplemented with 1.5% agar base layer (base layer), the addition that the absorbance is inoculated on the BHI medium 1% of the diluted bacterial solution 10% agar is added to a 0.1-over layers at OD ₅₅₀ ㎚ and then the size of the inhibitory ring was measured by the antimicrobial activity on an 8 ㎜ paper disk on which 2 ㎎ of 70% ethanol extract and hot water extract were respectively treated.

As a result, it was confirmed that when the culture plate containing the Streptococcus mutans strain at a certain concentration was treated with the extract of Chionoianaceae, a visible inhibitory ring was generated from the 70% ethanol extract to strongly inhibit the growth of the Chionae One).

Experimental Example  3. Glucan  produce Low performance  Measure

Glucosyltransferase secreted by Toxoplasma gonorrhepsis is collected using the property of glucosyltransferase which forms insoluble glucan on the tooth surface using sucrose in the food in the oral cavity as a substrate. The extract and its organic solvent fractions were added to confirm glucanogenesis inhibition. First, Streptococcus mutans strain was inoculated into BHI broth and cultured in a 37 ° C incubator for 24 hours, followed by centrifugation at 4,300 rpm for 30 minutes at 4 ° C to take a supernatant. The supernatant was filtered through a 0.2 쨉 m syringe filter, and then a crude enzyme solution was prepared using Amicon Ultra Centrifugal Filter (MWCO 30 kDa, Cat. No. UFC903008, Millipore).

The activity of glucosyltransferase was confirmed by measuring the concentration of insoluble glucan produced using sucrose as a substrate using a spectrophotometer. For this purpose, a substrate to the glass test tube, a solution (0.0625 M Potassium Phosphate Buffer (pH 6.5) 1 riteo of Sucrose 12.5 g, NaN ₃ 0.25 g contained) 800 ㎕, crude enzyme solution 20 ㎕ and jingyo hot water extract or solvent fraction or goldthread extract 180 ㎕ To make the final volume 1 ml. As a negative control, 180 [mu] l of distilled water was added. The test tube was incubated at 37 ° C for 30 hours while being inclined at about 30 °. After the reaction, the supernatant was discarded by centrifugation, 4 ml of distilled water was added, and ultrasonication was performed for 10 minutes to disperse the formed glucan. Immediately after dispersion, the absorbance at 550 nm was measured with a spectrophotometer, and the inhibition rate of each sample against glucosyltransferase was calculated by the following equation, and the results are shown in Table 2.

(%) = [(Control - control blank) - (sample - sample blank)} / (control - control blank)] × 100

Treatment concentration
(占 퐂 / ml) Hot water extract Ethyl acetate
Fraction Butanol
Fraction goldthread
Hot water extract 500 250 250 500 Glucan production
Inhibition rate (%) 64.59 ± 0.29 54.60 ± 0.64 34.93 + - 1.22 53.35 ± 1.28

As can be seen from Table 2, glucan production was most strongly inhibited in the hydrothermal extract-treated group in which the antimicrobial activity against the tooth root was weak. In the butanol fraction, the glucan production inhibitory activity was 30% or more. Especially, the ethyl acetate fraction showed similar inhibitory activity even at the dilution concentration twice as high as that of the hot - water extract. In addition, the antioxidative activities of the extracts of Huangyong hot - water extracts were compared with those of the extracts of Huangyong hot - water extracts.

Experimental Example  4. Determination of total phenolic compounds content

In the past, it has been reported that the content of phenolic compounds is high in herbal medicines known to have dental caries inhibition activity (Lee, HO, et al., Antibacterial effects of sophora flavescens on Streptococcus mutans. Korea J. Food Nutr. 13: 35 (3): 284-293, 2006). Based on the results, it was found that the extract from hot water and the hot water extract-derived organic solvent The content of total phenolic compounds in the fractions was determined using the Folin-Denis method (Slater TF. 1984. Free radical mechanisms in tissue injury. Biochem J 222: 1-15). 2 ml of hot water extract and two kinds of solvent fractions were diluted 10 times with distilled water, and then 2-fold diluted Folin-Ciocalteu 'reagent (Sigma, MO, USA) was added thereto for 3 minutes. Then, the same amount of 10% Na ₂ CO ₃ was slowly added and reacted for 1 hour. Absorbance was measured at 700 nm using a spectrophotometer (UV / Vis Spectrophotometer U-2900, HITACHI, Tokyo, Japan). From the measured absorbance, the amount of the total phenolic compound was converted into a standard curve using tannic acid, and the results are shown in Table 3.

Total polyphenol Concentration (/ / ml) Hot water extract 65.36 ± 1.73 Ethyl acetate fraction 293.18 ± 0.97 Butanol fraction 232.28 ± 5.33

As can be seen from Table 3, the phenolic compound content was the highest in the ethyl acetate fraction and the phenolic compound was present in the butanol fraction in a similar manner to the antimicrobial activity confirmed in the control Respectively. This suggests that the antimicrobial activity of chinchillas against the phytopathogenic fungi may be due to the phenolic compounds present in the chlorophyll.

Experimental Example  5. Acute toxicity  Experiment

Single - dose toxicity studies were conducted to obtain data on the safety of extracts from Chinchon. Single-dose toxicity tests were conducted using female and male ICR mice according to the "Nonclinical Test Control Standard" (Oct. 25, 2005) of Toxicological Test Methods for Drug and Other Notification Notification No. 2005-60 of the Food and Drug Administration. The temperature of 23 ± 3 ° C, relative humidity of 50 ± 10%, the number of times of ventilation 12 to 16 times per hour, illumination 12 hours of light intensity (light 7:00, light 19:00), illumination 150 ~ 300 Lx The animals were inspected and visualized under the environment of human rearing, and their appearance was visually examined. Male and female mice were set up in groups of 5 mice per group, and were housed individually in cages. Feeds and water were fed ad libitum and feeds were fed with animal feed (PMI nutrition, USA).

The dose of hot water extract of Jeungbo was set to 3 doses of 2,500 ㎎ / ㎏ and 1,250 ㎎ / ㎏ (Mouse, BW) with an air ratio of 0.5, based on 5000 ㎎ / ㎏. Were used for the test. The route of administration was oral administration. The animals were fasted overnight and then orally administered after fixation of the body using a disposable sonde and a suture tube for oral administration.

Clinical symptoms were observed every hour for 6 hours from the time of administration, and then, for the next 14 days, general symptoms such as change in general condition, mobility, appearance, autonomic nerve, and death were observed once a day. Body weights were measured at the time of arrival, before separation of the group and at 1, 3, 7, and 14 days after the test substance administration, respectively. The animals were fasted for 18 hours on the night before sacrifice and then anesthetized by intraperitoneal injection of Zoletil 50 (Virbac SA, France) and Xylazine Hydrochloride (Bayer Korea Co., Ltd., Kyonggi-do) And gross lesions were observed for all organs.

As a result, the administration of test substances such as gait disorder, behavioral abnormality, cough, diarrhea, edema, shortness of breath, grooming, jumping, gait, lethargy, vomiting, No clinically relevant abnormalities were observed, and no deaths were observed in all female / male ICR mice (Table 4). Thus, the extract is jingyo limit capacity (5,000 ㎎ / ㎏ / day) to not even be observed by oral administration in death LD ₅₀ No value was calculated.

group Volume
(Mg / kg) gender Number of animals Number of animals death rate(%) Control group 0 cock 5 0 0 female 5 0 0 Extract 1,250 cock 5 0 0 female 5 0 0 2,500 cock 5 0 0 female 5 0 0 5,000 cock 5 0 0 female 5 0 0

In addition, the body weight change of the experimental animals was observed for 1, 3, 7, and 14 days after oral administration of the hot water extract of Chinchilla japonica. As a result, it was found that all the experimental animals had more than 25% Increased by more than 10%. No significant weight loss or increase effect was observed in the experimental animals (Table 5).

group Volume
(Mg / kg) Weight (g) 0 days 1 day 3 days 7 days 14 days Control 1 0 Average 25.136 28.210 29.070 31.264 33.030 SD 1.984 2.271 2.008   2.419 0.466 cock
(male) 1,250 Average 24.964 27.108 27.958 30.334 32.234 SD 1.832 3.126 3.017 2.795 2.989 2,500 Average 25.030 27.404 28.378 30.738 32.568 SD 1.285 2.004 1.993 1.673 2.292 5,000 Average 24.974 28.446 29.326 30.872 33.114 SD 0.760 1.539 1.664 1.368 1.450 Control group 2 0 Average 22.008 21.308 23.382 23.458 24.932 SD 1.299 0.952 1.104 0.923 1.025 female
(female) 1,250 Average 21.928 22.626 22.692 24.568 26.830 SD 1.295 1.833 1.827 2.100 2.502 2,500 Average 21.890 22.988 22.916 22.532 23.832 SD 1.138 0.981 1.029 1.379 0.977 5,000 Average 21.694 21.836 22.304 23.434 25.070 SD 0.718 1.310 1.726 1.026 1.326

Manufacturing example  One: Whole life  Preparation of oral compositions containing extracts

<1-1> Manufacture of Toothpaste

Extract of the extract of Example 1 1 g &lt; tb &gt;

Sodium hydrogen carbonate ··································································· 20 g

Glycerin ········································· 15 g

White Clay · · · · · · · · · · · · · · · · · · · · · · · · · · · · · 2 g

Bamboo salt · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · ·

Vitamin E ······················· 2 g

Small amount to the viscosity of purified water

The above ingredients were mixed to prepare a natural toothpaste having a dental caries prevention effect. Menthol or peppermint essential oil may be added to give a cooling sensation.

<1-2> Manufacture of oral cleaning agent

The extract of goethite of Example 1 .................. 0.5%

Ethanol ····················· 15%

Glycerin · · · · · · · · · · · · 10%

Sodium benzoate &lt; SEP &gt; 0.15% &lt; tb &gt;

Sucarin sodium ····························· 0.1%

Peppermint essential oil ······················ 0.15%

Purified water (ion-exchanged water)

pH 6.0

Oral detergent having dental caries prevention and bad breath prevention effect was prepared by adding alcohol, wetting agent, ethanol, glycerin, and sodium benzoate as general oral detergent ingredients, and the extract prepared in Example 1. Peppermint essential oil may be added to add a cooling sensation, and sodium saccharin may be added to adjust the taste of the herb extract.

Manufacturing example  2: Whole life  Preparation of health food containing extract

<2-1> Manufacturing of food

Foods containing the extract of Shin-kyo as prepared in Example 1 were prepared as follows.

1. Preparation of cooking seasoning

The safflower for health promotion was prepared from 20 to 95% by weight of the extract.

2. Manufacture of tomato ketchup and sauce

0.2 ~ 1.0% by weight of the extract of Chinchilla japonica was added to tomato ketchup or sauce to prepare healthy tomato ketchup or sauce.

3. Manufacture of flour food

0.5 ~ 5.0% by weight of the extract of Chrysanthemum morifolium was added to wheat flour, and bread, cake, cookies, crackers and noodles were prepared by using this mixture to prepare foods for health promotion.

4. Manufacture of soups and gravies

0.1 ~ 5.0 wt.% Of the extract of Chinchilla japonica was added to the soup and the juice to prepare a health improvement meat product, noodle soup and juice.

5. Manufacture of ground beef

The ground beef for health promotion was prepared by adding 10% by weight of the extract from the ground beef.

6. Manufacture of dairy products

5 to 10% by weight of the crude extract was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.

7. Manufacturing of wires

Brown rice, barley, glutinous rice, and yulmu were dried by a known method and dried, and the mixture was granulated to a powder having a particle size of 60 mesh. Black soybeans, black sesame seeds, and perilla seeds were steamed and dried by a conventional method, and then they were prepared into powder having a particle size of 60 mesh by a pulverizer. The crude extract was concentrated under reduced pressure in a vacuum concentrator, sprayed, and dried with a hot-air drier, and the resulting dried product was pulverized to a particle size of 60 mesh with a pulverizer to obtain a dry powder. The above-prepared cereal grains, seeds and dried extract powder were prepared by blending the dried powders in the following proportions.

Cereals (30% by weight of brown rice, 15% by weight of yulmu, 20% by weight of barley)

Seeds (7% by weight of perilla, 8% by weight of black beans, 7% by weight of black sesame seeds)

The dried powder (3% by weight) of the crude extract,

(0.5% by weight),

(0.5% by weight)

<2-2> Production of beverage

1. Manufacture of carbonated beverages

Additive of 1 to 5% of the extract of Allium cepaense, 5 to 10% of sugar, 0.05 to 0.3% of citric acid, 0.005 to 0.02% of caramel and 0.1 to 1% of vitamin C was mixed and 79 to 94% of purified water was mixed therein to make syrup , The syrup was sterilized at 85 to 98 ° C for 20 to 180 seconds, mixed with cooling water at a ratio of 1: 4, and 0.5 to 0.82% of carbon dioxide gas was injected to prepare a carbonated drink containing the extract.

2. Manufacture of health drinks

(3%) and fructose extract (3%) such as liquid fructose (0.5%), oligosaccharide (2%), sugar (2%), salt (0.5% This was packaged in small containers such as glass bottles and PET bottles to produce health drinks.

3. Manufacture of vegetable juice

Healthy vegetable juice was prepared by adding 5 g of the extract of Chungbuk National University to 1,000 ㎖ of tomato or carrot juice.

4. Manufacture of fruit juice

1 g of the extract was added to 1,000 ml of apple or grape juice to prepare fruit juice for health promotion.

Claims (11)

A composition for oral use for preventing or treating dental caries comprising as an active ingredient an extract of Jin-pyo extract obtained by extracting Jin-bridge with water or an alcohol-extracting solvent. The method according to claim 1,
Wherein said extract is prepared using 2-30 parts by volume (v / w) of an extraction solvent. delete The method according to claim 1,
Wherein the alcohol is selected from the group consisting of C ₁ to C ₄ lower alcohols and aqueous solutions thereof. The method of claim 4,
Wherein the organic solvent is an aqueous solution of ethanol or ethanol. The method according to claim 1,
Wherein the extract is a fraction obtained by adding a fractionated organic solvent to a water extract of Jeung-shi. The method of claim 6,
Wherein the fractionated organic solvent is ethyl acetate or butanol. The method according to claim 1,
Wherein the composition comprises 0.01 to 50% by weight of the extract of Chrysanthemum morifolium according to the total weight of the oral composition. The method according to any one of claims 1, 2, and 4 to 8,
Wherein the oral composition is selected from the group consisting of dentifrices, mouthwashes, and oral sprayed formulations. A health food for preventing or alleviating dental caries comprising as an active ingredient an extract of Jin-kyo, which is obtained by extracting Jin-bridge with water or an alcohol-extracting solvent. The method of claim 10,
Wherein said health food is in the form of a food or beverage.

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