KR20120133134A - Composition for Prevention or Treatment of Dental Caries Comprising Extract of Tetrapanax papyriferus - Google Patents

Abstract

PURPOSE: A composition containing a Tetrapanax papyriferus extract for preventing or treating dental caries is provided to ensure antibacterial effect and to suppress glucosyltransferase activity. CONSTITUTION: A composition for preventing or treating dental caries contains 0.01-50 wt% of Tetrapanax papyriferus extract as an active ingredient. The extract is prepared using water or an organic solvent. The organic solvent is low alcohol of C1-C4, acetone, or a solution thereof. The extract is a fraction prepared by adding an organic solvent to the water extract of Tetrapanax papyriferus The oral composition is a toothpaste, oral cleanser, or oral spray. A health functional food for preventing or relieving dental caries contains the extract as an active ingredient. [Reference numerals] (AA) Tetrapanax papyriferus

Description

Composition for Prevention or Treatment of Dental Caries Comprising Extracts of Tobacco Herbal Medicine {{Composition for Prevention or Treatment of Dental Caries Comprising Extract of Tetrapanax papyriferus}

The present invention relates to a composition for preventing or treating dental caries, and more particularly, to a composition containing the extract of the herb as an active ingredient, which can be usefully used for the prevention and / or treatment of dental caries.

Dental caries is a disease in which the hard tissues of teeth are destroyed by the action of organic acids produced by the metabolism of carbohydrates in food by microorganisms in the gingival and subgingival bacterial membranes. It is one of the global concerns as well as Korea. Dental caries does not have a direct relationship to human life, but since the chewing function of food decreases, it directly or indirectly affects diseases of the digestive system. The morbidity rate of dental caries is particularly high in children, with 85% of children aged 5 years having experienced or experiencing pain from dental caries.

Although there are many causes of dental caries, lactic acid, which is caused by the breakdown of carbohydrates such as sugar and starch of food residues attached to teeth by the fermentation of bacteria that resides in the oral cavity, desorbs the lime of the dental hard tissues. Will occur. Several species are known as pathogenic microorganisms that cause dental caries, but in particular Streptococcus mutans ), Streptococcus sobrinus ) and the like act as a major pathogen for the development of dental caries (Non-Patent Document 1). Streptococcus mutans secrete an enzyme called glucosyltransferase (GTase) to extracellular or cell surface to produce glucose and fructose based on sucrose in food, and at the same time insoluble glucan, a polymer of glucose ) Will form on the surface. This glucan causes other microorganisms in the oral cavity to adhere to the dental plaque, forming a dental plaque, or dental plaque. Within this plaque, lactic acid bacteria, including Streptococcus mutans bacteria, multiply to produce lactic acid as a carbon source, and lactic acid is collected 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 used to suppress such dental caries include methods using antibiotics such as sporamycin, vancomycin, and chlorohexidine, and methods of inhibiting pathogenic bacteria by organic or inorganic fluoride. Etc. are used (Non Patent Literature 3). However, if the antibiotics are used continuously, not only the microorganisms become resistant or cause side effects such as diarrhea and vomiting, but also the problem of inhibiting the growth of bacteria in the oral cavity due to the abuse of these antibiotics. have. Therefore, there is an urgent need for the development of effective and stable prevention and treatment of dental caries.

On the other hand, Tongcho (通 草, Tetrapanax papyriferus K. ) refers to the stems of the tree (Tetrapanax papyriferus K. Koch), the appearance is columnar, white, the quality is light and flexible, hollow in the middle, elastic There is this and is easy to be broken. Tongcho extends into vines with the same thickness as fingers, with 2-3 branches per node. Five leaves hang at the end of the branch and bear fruit, similar to small hairs. The seed is black and the inside is white, but it is sweet because it is sweet. Lunar New Year, in February, the branches are dried in the shade, and there is a hole in the stem, which leads to both ends.

Tongcho is named because the urethra is blocked because the urine does not look cool, and there are several species of water (水腫). Tongcho has no smell, taste is sweet, and it is slightly cold. In addition, Tongcho lowers the heat and increases the amount of urine to promote gonorrhea, urethritis, cystitis, postpartum milk secretion. In addition, according to the herbology, Tongcho heals five kinds of disease, urinates well, releases physiognomy, heals species, and improves heat, ) To work well. Currently, it is known that the extract of Tongcho has anticancer activity against human oral cancer cell lines (Non-Patent Document 4), and there are reports that Tongcho may be used as a component of the herbal composition for lowering blood sugar (Patent Document 1 and Patent Document 2). However, no literature has been reported that can be used to prevent or treat dental caries.

Accordingly, the present inventors have studied various herbal extracts that can be usefully used for the prevention or treatment of dental caries. As a result, the present inventors have shown that the extracts of the herbaceous plants exhibit excellent dental caries prevention or treatment effects that have not been reported before. The invention has been completed.

Republic of Korea Patent Registration No. 10-0827350 Republic of Korea Patent Publication No. 10-2004-0026276

 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  Jeong, Sun-Hee, "The Effect of Tobacco Extract on Human Oral Cancer KB Cell Line", Thesis, Chosun University, 2010

It is an object of the present invention to provide a composition for the prevention or treatment of dental caries, which contains the herb extract as an active ingredient.

In order to achieve the above object, the present invention provides a composition for the oral cavity for the prevention or treatment of dental caries comprising the extract as an active ingredient.

In addition, the present invention provides a health food for the prevention or alleviation of dental caries containing the herb extract as an active ingredient.

The present invention provides a composition for the oral cavity for the prevention or treatment of dental caries containing the herb extract as an active ingredient.

The Tongcho extract is preferably extracted by adding an extraction solvent of 2 to 30 times, preferably 5 to 20 times, to the Tongcho. Although water is generally used as the extraction solvent, other organic solvents may be used in addition to water as necessary. Organic solvents that may be used include, but are not limited to, C ₁ to C ₄ lower alcohols, preferably ethanol, acetone, or aqueous solutions thereof. In the present invention, the "tobacco extract" is used to encompass not only the tobacco extract prepared using the extracting solvent, but also the fraction extracted by adding one or more "fractionated organic solvents" to the tobacco extract. The fractionated organic solvent is not particularly limited, and conventional organic solvents may be used without limitation. In a preferred embodiment of the present invention, an organic solvent such as ethyl acetate or butanol was used as the fractionated organic solvent with respect to the water extract of the herb.

The composition for oral cavity containing the tobacco extract of the present invention may be prepared by using a formulation such as toothpaste, mouthwash or mouth spray, but is not limited thereto. In the composition for oral cavity of the present invention, the toothpaste may include one or more components such as abrasives, binders, moisturizers, foaming agents, sweeteners, flavoring agents and the like. Examples of the abrasive include aluminum hydroxide, silicic anhydride, aluminum silicate, dicalcium phosphate, dihydrate and anhydride, tricalcium phosphate, calcium carbonate, calcium pyrophosphate, insoluble sodium metaphosphate, magnesium triphosphate, magnesium carbonate, and sulfuric acid. One, two or more of calcium, polymethyl methacrylate, and other synthetic resins may be used in combination in an amount of about 20 to 90% of the total composition (20 to 60% for paste toothpaste). Examples of the binder include synthetic resins such as garginine, various thickening cellulose derivatives, xanthan gum, gums such as tragacanth gum, polyvinyl alcohol, sodium polyacrylate, polyacrylic acid / maleic acid copolymer, and carboxyvinyl polymer. In organic binders, such as a polymeric derivative, and inorganic binders, such as a silica and a laponite, 1 type, or 2 or more types can be mix | blended in the quantity of about 0.3-5% of the whole composition. Paste and liquid oral compositions in dentifrices include sorbitol, glycerin, ethylene glycol, propylene glycol, polyether glycol, and polypropylene glycol as moisturizing agents in the preparation of about 10-70% of the total composition. It can be used in combination.

On the other hand, the composition for oral cavity containing the herbal extract of the present invention is menthol (Mentol), anetol (Anetol), Eugenol (Eugenol), limonene (Limonene), Ocimene, n-dodecyl alcohol (n -Dodecy Alcohol, Citronenol, Alpha-Terpineol, α-Terpineol, Methyl salycilate, Methyl Acetate, Citroneryl Acetate, Cineol ), Linalol, Ethyl Linalool, Vanillin, Thymol, Spearamint Oil, Sage Oil, Rosemary Oil, Cinnamon Oil These and other pharmaceuticals may be used alone or in combination in an amount of 0.1 to 10%, preferably 0.5 to 5% of the total composition. In addition, sweeteners such as sodium saccharin, stevioside, aspartame, D-Xylose and the like can be used in combination in an amount of 0 to 1%, preferably 0.01 to 0.5% of the total composition.

In addition, the composition for oral cavity of the present invention includes alkali metal monofluoro, such as cationic fungicides such as triclosan, chlorohexidine, benzetonium chloride, benzalkonium chloride, cetylpyridium chloride, sodium monofluorophosphate and potassium monofluorophosphate Fluorides such as phosphate, sodium fluoride, and tin fluoride, tranexamic acid, epsilon amino capronic acid, aluminum chlorohydroxy allantoin, dihydrocholestanol, cletilychinates, glycerophosphates, sodium chloride, and water-soluble inorganic phosphate compounds And one or two or more of various efficacy components such as ascorbic acid (sodium), dl-a-tocopherol acetate, xylitol, pyridoxine hydrochloride and polyvinyl pyrrolidone can be used in combination.

In addition, anionic surfactants, nonionic surfactants and / or cationic surfactants may be used as the surfactant used in the composition for oral cavity of the present invention. Here, the anionic surfactants include sodium lauryl sulfate, sodium alkyl sulfates such as myristyl sulfate, sodium N-Lauroyl sarcosinate, sodium N-miristyl sarcosinate, and sodium N-miristyl sarcosinate. Sodium N-acyl sarcosinate and the like can be used, and non-ionic surfactants include sugar-derived fatty acid esters such as sugar fatty acid esters, maltose fatty acid esters, lactose fatty acid esters, and polyethers. Oxyethylene hardened castor oil, polyoxyethylene higher alcohol ether, polyoxy ethylene polyoxy propylene copolymer, polyoxy ethylene polypropylene fatty acid ester and the like can be used. Cationic surfactants include N-alkyl di aminoethylglycine, such as N-lauryl di aminoethylglycine, N-myristyl diaminoethylglycine, N-alkyl-N-carboxymethyl ammonium betaine, 2-alkyl-1 Hydroxyethyl imidazoline betaine sodium and the like can be used. In this case, the anionic surfactant is mainly used as the foaming component, and sodium alkyl sulfate such as sodium lauryl sulfate is most widely used. Moreover, it is preferable to use these surfactant individually by 1 type or in mixture of 2 or more types, and, as for a compounding quantity, 0.01-0.5%, More preferably, 0.05-0.3% of the whole composition is used normally.

The tobacco extract may be included in an amount of 0.01 to 50% by weight based on the total weight of the composition for oral cavity of the present invention, the content of which may be appropriately adjusted according to the absorption of the active ingredient, the age, sex, condition, degree of dental caries of the patient, etc. Can be.

According to one embodiment of the present invention, the herb extract exhibits strong antibacterial activity against the causative agent of dental caries, and particularly, shows strong anti-fungal bacterium activity in the ethyl acetate fraction of the herb extract (see Table 1). In addition, in the antimicrobial activity experiment using the disk diffusion method, visible inhibition ring is observed in 70% ethanol extract, it can be confirmed that strongly inhibit the growth of dental caries (see Fig. 1).

In addition, according to another embodiment of the present invention, glucan production is most strongly inhibited in the hydrothermal extract treatment group in which the antibacterial activity against dental caries is weak, while in the ethyl acetate and butanol solvent fractions, about half of the glucan is extracted. In particular, ethyl acetate fraction showed a low inhibitory activity, especially in the ethyl acetate fraction which showed strong antibacterial activity against dental caries. The antibacterial activity of the ethyl acetate fraction showed a direct inhibition of bacterial growth rather than glucosyltransferase activity. It can be inferred that it is represented by (see Table 2).

In addition, according to another embodiment of the present invention, the content of the phenolic compound in the ethyl acetate fraction was determined to be the highest in the ethyl acetate fraction, similar to the degree of antimicrobial activity observed in dental caries, and a large amount of the phenolic compound was present in the butanol fraction. It can be confirmed, which suggests that the antibacterial activity against the dental caries of the herb may be due to the phenolic compound present in the herb (see Table 3).

In addition, according to another embodiment of the present invention, as a result of toxicity experiments using mice, no abnormal symptoms of any clinical symptoms related to administration of the test substance were observed in all mice used in the experiment, and no dead animals were also observed (Table 4 Reference). together. All experimental animals showed no significant weight loss or gain (see Table 5).

In addition, the present invention provides a health food for the prevention or alleviation of dental caries containing the herb extract as an active ingredient.

The tobacco extract may be added to health foods for the purpose of preventing or improving dental caries symptoms, and when the tobacco extract is used as a food additive, it may be added as it is or used with other foods or food ingredients, Can be used accordingly. 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, in the preparation of food or beverages, the tobacco extract is added in an amount of 30% by weight or less, preferably 10% by weight or less 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 substance can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, teas, drinks, Alcoholic beverages and vitamin complexes, and the like and include all of the health foods in the 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 glucose, monosaccharides such as fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, 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 tobacco extract.

In addition to the above, the health beverage composition of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, Alcohol, a carbonation agent used for carbonated drinks, and the like. In addition, the tobacco extract may contain the flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components can be used independently or in combination. The ratio of such additives is not critical, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of tobacco extract.

As described above, the tobacco extract has an excellent antibacterial effect on dental caries, inhibits the activity of glucosyltransferase, an enzyme released during the growth and growth of dental caries, and its effect is also persistent. In addition, since it is very safe to be extracted from natural vinegar, it can be usefully used as a composition for oral health or health food for preventing and / or treating dental caries.

1 is a photograph showing the growth inhibitory experimental results showing the antimicrobial activity of the tobacco extract against dental caries, (A) shows that growth inhibition ring of 21 mm was formed upon administration of 2 mg of 70% ethanol extract, (B) Shows that growth inhibition ring of 10 mm was formed upon administration of 2 mg of hydrothermal extract.

Hereinafter, the present invention will be described in detail by way of examples.

However, the following examples are only for illustrating the present invention, and the content of the present invention is not limited by the following examples.

Example  One: Head  Preparation of extract

<1-1> Heat number  Preparation of extract

1 liter of distilled water was added to 50 g of dried tobacco, followed by extraction with hot water for 3 hours, followed by filtration and lyophilization using a filter paper.

<1-2> Preparation of organic solvent extract

0.5 g of 70% ethanol was added to 50 g of dried tobacco, sonicated at 25 ° C. for 1 hour, filtered using filter paper, and the solvent was removed using a rotary concentrator under reduced pressure.

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

After suspending the hot water extract of Tongcho prepared in Example <1-1> in water, and using ethyl acetate and butanol in consideration of the polarity of the solvent, the system fractionation from the low polar solvent to a high polar solvent sequentially An organic solvent fraction was obtained. The obtained organic solvent fractions were concentrated using a vacuum rotary concentrator, and then dissolved in DMSO (Dimetyl Sulfoxide) at a concentration of 25 mg / mL and stored at -20 ° C.

Comparative example  One. goldthread  Preparation of extract

Except for using the antibacterial activity of the dental caries have been reported in place of the tobacco bark in the same manner as in Example <1-1> was prepared by extracting the water.

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

The Streptococcus mutans strain, the causative agent of dental caries used in this experiment, is a Gram-positive and anaerobic bacterium. The KCTC3065 strain, which was distributed from the Korea Institute of Bioscience and Biotechnology, was isolated from BHI broth (Difco, USA) Subcultured in Agar medium was used for the experiment. In order to investigate the growth inhibitory activity of the tobacco extract against the causative agent of dental caries, the Streptococcus mutans strain was inoculated with BHI broth and passaged, and 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 μl aliquoted in 96-well microplates. After dispensing, the final concentration was added to the wells such that the total concentration of 70% ethanol extract and hot water extract was 1.25 mg / ml, and the organic solvent fraction derived from hot water extract was 0.625 mg / ml. After incubation for 20 hours, the absorbance was measured to measure growth inhibition, and the results are shown in Table 1.

Treatment concentration
(Μg / ml) 70% ethanol
extract Hot water extract Ethyl acetate
Fraction Butanol fraction 1,250 1,250 625 625 Dental caries
Growth inhibition (%) 66.79 ± 0.43 10.00 ± 0.62 48.21 ± 0.81 14.94 ± 0.75

As can be seen from Table 1, the strongest antimicrobial activity was confirmed in 70% ethanol extract. In addition, when the ethyl acetate fraction of the water extract was treated, anti-fungal bacterium activity was confirmed about 4 times stronger even at twice the dilution concentration than the hot water extract of the mother liquor Tongcho, and the active ingredient was concentrated by the solvent fraction to increase the efficiency. It was confirmed.

Experimental Example  2. Determination of antimicrobial activity against bacteria causing dental caries

The antimicrobial activity of the tobacco extract prepared in Example 1 was measured by the disk diffusion method. To this end, a BHI plate containing 1.5% agar was inoculated into a base layer, and 10% of the bacterial solution diluted to an absorbance of 0.1 at ₅₅₀ nm in BOD medium containing 1% agar. After the preparation (over layer), the size of the inhibitory ring due to antimicrobial activity was measured by placing 8 mm paper disks treated with 2 mg of 70% ethanol extract and hot water extract.

As a result, as a result of treatment of the tobacco extract to the culture plate containing a certain concentration of Streptococcus mutans strain, it was confirmed that visible inhibition ring is generated in 70% ethanol extract to strongly inhibit the growth of dental caries (FIG. One).

Experimental Example  3. Glucan  produce Low performance  Measure

Using glucosyltransferase which forms insoluble glucan on the tooth surface by using sucrose in oral food as a substrate, glucosyltransferase secreted by dental caries is collected and buffer solution containing sucrose and whole hot water Extracts and their organic solvent fractions were added to confirm glucan production inhibition. First, Streptococcus mutans strains were inoculated in BHI broth and incubated in a 37 ° C. incubator for 24 hours, followed by centrifugation at 4,300 rpm for 30 minutes to obtain supernatant. After filtering the supernatant with a 0.2 μm syringe-type filter, a crude enzyme solution was prepared using an 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, 800 μl of substrate solution (containing 12.5 g of Sucrose, 0.25 g of NaN _{3 in} 1 liter of 0.0625 M Potassium Phosphate Buffer (pH 6.5)), 20 μl of coenzyme solution and 180 μl of hot water extract or solvent fraction or rhubarb extract Was added to bring the final volume to 1 ml. As a negative control, 180 μl of distilled water was added. The test tube was inclined at about 30 ° and allowed to react for 30 hours in a 37 ° C incubator. After the reaction, the supernatant was discarded by centrifugation, and 4 ml of distilled water was added to sonicate for 10 minutes to disperse the formed glucan. The absorbance was measured at 550 nm with a spectrophotometer immediately after dispersion, and the inhibition rate for glucosyltransferase of each sample was calculated by the following formula, and the results are shown in Table 2.

% Inhibition of glucan production = [{(control-control blank)-(sample-sample blank)} / (control-control blank)] × 100

Treatment concentration
(Μg / ml) Hot water extract Ethyl acetate
Fraction Butanol
Fraction goldthread
Hydrothermal extract 500 250 250 500 Glucan generation
Inhibition rate (%) 47.46 ± 4.12 20.94 ± 3.96 27.02 ± 1.51 53.35 ± 1.28

As can be seen from Table 2, the glucan production was most strongly inhibited in the hydrothermal extract treatment group in which the antibacterial activity against dental caries was weak. On the other hand, the two solvent fractions showed about 20% inhibition of glucan production. In particular, the ethyl acetate fraction, which exhibited strong antibacterial activity against dental caries, exhibited relatively low inhibition of glucan production. It could be inferred that this was indicated by direct fungal growth inhibition rather than siltransase activity inhibition. In addition, as a result of comparing the activity after the same amount of the antibacterial activity against the dental caries, which was reported the antibacterial activity, it was confirmed that the tobacco extract and the extract of the yellow lotus showed similar inhibition rate of glucan production.

Experimental Example  4. Determination of total phenolic compound content

It has been reported that the content of phenolic compounds is high in herbal medicines known to have inhibitory activity against dental caries (Lee HO, et al., Antibacterial effects of sophora flavescens on Streptococcus mutans.Korea J. Food Nutr. 13: 536-546, 2000; Yun-Mi Park, et al., J. Korean Soc.Food Sci.Nutr. 35 (3): 284-293, 2006), based on this 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). The hot water extract and the two kinds of solvent fractions were diluted 10 times with distilled water, and each of 2 ml was taken. The same amount of 2 times diluted Folin-Ciocalteu 'reagent (Sigma, MO, USA) was added and reacted for 3 minutes. Thereafter, the same amount of 10% Na ₂ CO ₃ was slowly added and reacted for 1 hour, and the absorbance was measured at 700 nm using a spectrophotometer (UV / Vis Spectrophotometer U-2900, HITACHI, Tokyo, Japan). From the measured absorbance, the total amount of the phenolic compound was converted into a standard curve using tannic acid, and the results are shown in Table 3.

Total polyphenol Concentration (µg / ml) Hot water extract 80.49 ± 1.24 Ethyl acetate fraction 128.18 ± 1.94 Butanol fraction 103.31 ± 1.02

As can be seen from Table 3, the content of phenolic compounds in the ethyl acetate fraction was measured to be the highest in the ethyl acetate fraction, similar to the degree of antibacterial activity observed in dental caries, and the large amount of phenolic compounds were also present in the butanol fraction. Confirmed. This suggests that the antimicrobial activity of Tobacco on dental caries may be due to the phenolic compounds present in Tobacco.

Experimental Example  5. Acute toxicity  Experiment

A single dose toxicity test was performed to prepare data on the safety of the tobacco extract. Single-dose toxicity experiments were performed using male and female ICR mice according to the "Non-clinical Trial Management Standards" (October 25, 2005) of Toxicology Test Methods, such as Food and Drug Administration Notification No. 2005-60. About 1 week after purchase, temperature 23 ± 3 ℃, relative humidity 50 ± 10%, ventilation frequency 12 ~ 16 times per hour, lighting 12 hours light cycle (light up 7:00, light off 19:00), illumination 150 ~ 300 Lx The animals were visually inspected while being quarantined and purified under a human breeding environment, and healthy animals were selected for the experiment. Male and female mice were set up in 5 groups in each group, and 5 were individually housed in cages. Feed and water were freely ingested, and the feed was fed solid animal feed (PMI nutrition, USA) for laboratory animals.

Two doses of Thucho hot-water extract were set at 2,500 mg / kg and 1,250 mg / kg (Mouse, B.W.), and each of them was diluted in each dose with an excipient physiological saline and used for the test. The route of administration was administered by oral administration. After fasting the animals overnight, the body was fixed by the dorsal skin fixation method using oral administration disposable sonde and injection tube and then orally administered.

Clinical symptoms were observed every hour for 6 hours immediately after administration, and then general symptoms such as changes in general condition, motility, appearance, autonomic nerve, and death were observed once daily for 14 days. Body weights were measured at day 1, 3, 7, 14 after group separation and at the time of administration of test substance, respectively. After 18 hours of fasting the night before the sacrifice, the animals were anesthetized by intraperitoneal injection of Zoletil 50 (Virbac SA, France) and xylazine hydrochloride (Bayer Korea Co., Ltd., Gyeonggi-do). Afterwards, gross lesions on all organs were observed.

As a result, all the male and female ICR mice used in the experiment were treated with test substance such as gait disorder, behavioral disorder, squat, diarrhea, swelling, tightness of breath, grooming, jumping, seizures, lethargy, vomiting, nasal secretion, paralysis, and softness. Abnormal findings of any relevant clinical symptoms were not observed and no deaths were observed in all male and female ICR mice (Table 4). Therefore, even if the Tobacco extract is administered orally up to the limit dose (2,500 mg / kg / day), death can not be observed LD ₅₀ The value was not calculated.

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

In addition, as a result of oral single-dose administration of the extract, the body weight of the experimental animals was observed for 1, 3, 7, and 14 days. Showed a tendency to lose weight at, but recovered from day 7. No significant weight loss or increase was observed in all experimental animals (Table 5).

group Volume
(Mg / kg) Weight (g) 0 days 1 day 3 days 7 days 14 days Control group 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 25.054 26.610 27.492 28.956 31.520 SD 1.790 2.053 2.036 2.253 1.991 2,500 Average 24.932 24.678 25.500 27.044 29.660 SD 1.242 0.998 0.981 0.944 1.079 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 21.980 22.718 22.55 22.340 24.302 SD 1.122 1.003 1.003 1.103 1.571 2,500 Average 21.910 22.258 24.85 23.552 25.380 SD 0.837 0.748 0.748 1.235 2.372

Manufacturing example  One: Head  Preparation of the composition for oral cavity containing the extract

<1-1> Preparation of Toothpaste

Herb extract of Example 1 ?????????????????? 1 g

Sodium bicarbonate ???????????????????????? 20 g

glycerin????????????????????????? 15 g

White Clay ??????????????????????? 2 g

Bamboo salt ??????????????????????????? 10 g

Vitamin E ????????????????????????? 2 g

Small amount to match the purified water viscosity

By combining the above raw materials to prepare a natural toothpaste having a dental caries prevention effect. You can also add menthol or peppermint essential oils for a refreshing feel.

<1-2> Preparation of Oral Cleaning Agent

Herb extract of Example 1 ?????????????????? 0.5%

ethanol??????????????????????????? 15%

glycerin?????????????????????????? 10%

Sodium benzoate ???????????????????? 0.15%

Zalcarin Sodium ??????????????????????? 0.1%

Peppermint Refinery ?????????????????????? 0.15%

Purified Water (Ion Exchange Water) ??????????????????? 74.09%

pH 6.0

Alcohol, humectant, surfactant, ethanol, glycerin, sodium benzoate are used as a general mouthwash component, and the herbal extract prepared in Example 1 was added to prepare a mouthwash having an effect of preventing dental caries and bad breath. Peppermint essential oil can be added to add a refreshing feeling, and saccharin sodium can be added to adjust the taste by the herbal extract.

Manufacturing example  2: Head  Preparation of Health Foods Containing Extract

<2-1> Manufacturing of food

The food containing the tobacco extract prepared in Example 1 was prepared as follows.

1. Preparation of cooking seasoning

A health promotion cooking seasoning was prepared with 20-95% by weight of Tongcho extract.

2. Manufacture of tomato ketchup and sauce

Health-promoting tomato ketchup or sauce was prepared by adding 0.2-1.0% by weight of the tobacco extract to tomato ketchup or sauce.

3. Manufacture of flour food

Tobacco extract 0.5-5.0% by weight was added to the flour, using this mixture to prepare bread, cakes, cookies, crackers and noodles to produce health foods.

4. Manufacture of soups and gravies

Tonic extract 0.1-5.0% by weight was added to soups and gravy to prepare health products, noodle soup and gravy.

5. Manufacture of ground beef

10% by weight of the tobacco extract was added to the ground beef to prepare a ground beef for health promotion.

6. Manufacture of dairy products

5-10% by weight of Tobacco extract was added to milk, and the milk was used to prepare various dairy products such as butter and ice cream.

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 extract was concentrated under reduced pressure in a vacuum concentrator, dried by spraying and a hot air dryer, and ground to a particle size of 60 mesh using a grinder to obtain a dry powder. It was prepared by combining the dry powder of the grains, seeds and tobacco extract prepared in the following ratio.

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

Seeds (7% by weight perilla, 8% by weight black bean, 7% by weight black sesame)

Dry powder (3% by weight) of the soybean extract,

(0.5% by weight),

(0.5% by weight)

<2-2> Production of beverage

1. Manufacture of carbonated beverages

Add 1 ~ 5% of Tobacco Extract, 5 ~ 10% Sugar, 0.05 ~ 0.3% Citric Acid, 0.005 ~ 0.02% Caramel, 0.1 ~ 1% Vitamin C, and mix it with 79 ~ 94% Purified Water to make syrup. , The syrup was sterilized at 85-98 ° C. for 20-180 seconds, mixed with cooling water at a ratio of 1: 4, and 0.5-0.82% of carbon dioxide was injected to prepare a carbonated beverage containing a tobacco extract.

2. Manufacture of health drinks

Instant sterilization by homogeneously mixing subsidiary ingredients such as liquid fructose (0.5%), oligosaccharide (2%), sugar (2%), salt (0.5%), water (75%), and tobacco extract (3%) This was packaged in small packaging containers such as glass bottles and PET bottles to prepare a healthy beverage.

3. Manufacture of vegetable juice

Tobacco extract 5 g was added to 1,000 ml of tomato or carrot juice to prepare vegetable juice for health promotion.

4. Manufacture of fruit juice

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

Claims (11)

Oral composition for the prevention or treatment of dental caries containing tongcho extract as an active ingredient. The method according to claim 1,
Tongcho extract is a composition for oral cavity extracted by adding 2 ~ 30 times the amount of the extraction solvent to Tongcho. The method according to claim 2,
The extraction solvent is water or an organic solvent composition for oral use. The method according to claim 3,
The organic solvent is a composition for oral cavity selected from the group consisting of C ₁ to C ₄ lower alcohol, acetone and an aqueous solution thereof. The method of claim 4,
The organic solvent is an oral composition of ethanol or ethanol aqueous solution. The method according to claim 1,
The tongcho extract is a composition for oral cavity which is a fraction extracted by adding a fractionated organic solvent to the water extract of Tongcho. The method of claim 6,
The fractionated organic solvent is ethyl acetate or butanol composition. The method according to claim 1,
The composition for oral cavity, characterized in that containing the Tongcho extract 0.01 to 50% by weight based on the total weight of the composition for oral cavity. The method according to any one of claims 1 to 8,
The oral composition is selected from the group consisting of toothpaste, mouthwash and mouth spray. Health food for the prevention or alleviation of dental caries containing tong extract as an active ingredient. The method of claim 10,
The health food is a health food in the form of food or drink.

Images