KR101156623B1 - Antimicrobial agent, composition and food products for oral cavity - Google Patents

Abstract

The present invention relates to an antimicrobial agent, an oral composition and a food or drink including the same, more specifically, an antibacterial agent containing the extract as an active ingredient, wherein the wax apple or water apple extract exhibits antimicrobial activity specifically to bacteria in the oral cavity, It relates to a composition for oral cavity and food and drink comprising the same.

Description

Antimicrobial agent, composition and food products for oral cavity comprising same {Antimicrobial agent, composition and food products for oral cavity}

The present invention relates to an antimicrobial agent, oral composition and food and drink comprising a wax or water apple extract.

Some bacteria in the oral cavity may cause bad breath, periodontal disease, caries, etc., and it is necessary to kill them or suppress or inhibit their proliferation.

The main component of bad breath is volatile sulfur compounds (VSC) such as hydrogen sulfide and methyl mercaptan. VSC is caused by breakdown of proteins constituting exfoliated epithelial cells, leukocytes, etc. by bacteria in the oral cavity (first dental publication, quarterly dental care 20 (1) 31-39, 2006).

Periodontal disease refers collectively to lesions expressed in periodontal tissue. Periodontal disease is usually caused by abnormal growth of plaque (germ mass) in the teeth between the teeth or alveolar, causing inflammation. Dental home exudate, accompanied by periodontal disease, is also a cause of bad breath (published by Medicinal Publishing Co., Ltd., war against oral biofilm dental plaque bacteria, 135-137). Caries, also known as tooth decay, are caused by demineralized hemorrhoids by acids produced by bacteria in the oral cavity.

Many of the bacteria in the oral cavity live in plaques. Plaque, also known as dental plaque and tartar, is a structure mainly composed of bacteria formed on the surface of the tooth and its metabolites, and it is known that there are 10 ¹¹ bacteria per gram. Plaque removal is essential for the prevention, cure of bad periodontal disease, and prevention and reduction of bad breath.

Until now, chlorhexidine and cetylpyridinium chloride are known as an oral antibacterial agent. However, these antimicrobial agents have a broad antimicrobial spectrum, and as a result of this use, there is a concern that adverse effects may occur such that mycelial growth occurs and abnormal drug propagation occurs. Oral mycelial disease is known as oral candidiasis. Oral candidiasis is caused by overgrowth of fungi (fungi) called Candida albicans in the oral cavity. In addition, side effects are known for chlorhexidine.

In view of the above, as an antimicrobial agent having high safety, an antimicrobial agent having a specific effect on bacteria in the oral cavity and particularly exhibiting antimicrobial activity on bacteria causing bad breath, periodontal disease and tooth decay is desired. In addition, in order to use the antibacterial agent in the composition for oral cavity, and also the product which can be taken orally, the antimicrobial agent is preferably derived from a material having high safety and good image.

The wax apple (Syzygium samaragense) and the water apple (Syzygium aquaeum) are plants of the genus Myzaceae (Syzygium). Wax apples are cultivated in Taiwan, India, and the Philippines, and the fruits are used for apples with the same taste as apples (Published by Agricultural Books Co., Ltd., published by Miyao Nakamura, 378-379). Water apples are distributed and cultivated throughout tropical Asia (published by the Agricultural and Tropical Tropical Research Center, Fruit Trees in Southeast Asia, 394-400).

Fitoterapia 69 (4) 365-367, 1998 shows that the extracts of the bark of bark apples and the 95% ethanol extracts have antibacterial activity against Staphylococcus aureus, Streptococcus pyogenes, etc. Has been disclosed. However, the study was not based on oral bacteria. Staphylococcus aureus is a Staphylococcus aureus that is present in the skin and causes purulent diseases. Streptococcus piogenes is a purulent streptococci present in the skin and sometimes reproduces in the pharynx, but is not a bacterium closely related to plaque formation or oral disease. In addition, the lipase inhibitory activity of wax apples (Japanese Patent Laid-Open No. 2005-60334, WO 2005-056031) is known, and steam distillate can be used as a cosmetic (Japanese Patent Laid-Open No. 2001-220312). It is also known that water distillates can be used as cosmetics for water apples (Japanese Patent Laid-Open No. 2001-220312).

As antimicrobial agents derived from natural products, the following are known. Japanese Patent Laid-Open No. 2002-114658 discloses that a legume plant has an antimicrobial activity against bacteria in the oral cavity. In addition, Japanese Patent Application Laid-Open No. 58-39615 discloses the combination of yellow wall, cloves, eucalyptus, mace, celery, parsley extract, and sodium alkyl sulfate, exhibits an antibacterial effect against oral bacteria.

Accordingly, the present inventors have steadily studied, the extract of the wax apple or water apple has an antimicrobial activity, and also found that the antimicrobial activity is specific for bacteria in the oral cavity and completed the present invention.

Accordingly, it is an object of the present invention to provide an antimicrobial agent that exhibits antimicrobial activity specifically to bacteria in the oral cavity and a composition for oral cavity and food and drink including the same.

The present invention is characterized by an antimicrobial agent comprising a wax or water apple extract.

The present invention also includes the antimicrobial agent exhibiting activity against bacteria in the oral cavity.

In addition, the present invention includes the antimicrobial agent wherein the extract is an extract of the leaves of wax or water apple.

In addition, the present invention includes a composition or food or drink for oral cavity containing the antimicrobial agent.

The antimicrobial term as used herein refers to the inhibition or inhibition of the killing or proliferation of microorganisms including bacteria, and includes concepts such as sterilization, sterilization, sterilization, and disinfection.

The site of the wax or water apple is not specified, and leaves, stems, bark, branches, pillars, roots, flowers, buds, seeds, fruits, or rinds may be used, and more preferably leaves. Examples of bacteria in the oral cavity include the following. That is, as the bacterial formate fatigue related to bad breath Pseudomonas long eccentricity less (Porphyromonas gingivalis ), Fusobacterium nucleatum , Prevotella intermedia ), Prevotella loescheii ), Treponema denticola ), Porphyromonas endodontalis ), Peptococcus anaerobics anaerobius ), Micros Priboti ( Micros prevotii ), Eubasterium limosum ), Centipedia periodontii , Selenomonas Arterimidis arterimidis ), Fusobacterium periodonticum ), Eubacterium spp.), Bacteroides spp., and the bacterium related to periodontal disease, Porphyromonas gingivalis gingivalis ), Fusobacterium nucleatum ), Actinobacillus actinomycetemcomitans ), Campylobacter rectus , Tennerella forsythia ), Treponema denticola ), Prevotella intermedia ), Actinomyces Viscosos ), Streptococcus mutans ) and Streptococcus sobrinus .

Among these bacteria, in particular Porphyromonas gingivalis ) and antibacterial agents that are active against Fusobacterium nucleatum are thought to be effective in reducing oral bacteria.

Porphyromonas gingivalis is a Gram-negative anaerobic bacterium that is representative of periodontal causative bacteria and is known as a causative agent of bad breath by producing VSC. Porphyromonas gingivalis has strong adhesion and co-aggregates with a plurality of fungal species (published first dental publication, quarterly dental care 20 (1) 31-39, 2006). In addition, porphyromonas gingivalis produces proteases such as collagenase and gingifen, and superoxide dismutase, thereby inhibiting the host's immune response. It is also reported that Ginaffein, which is secreted by Porphyromonas gingivalis, contributes to the coagulation of Actinomyces viscous.

Peugeot bacterium nucleatum is also a Gram-negative anaerobic bacterium and is known to be a causative agent of periodontal disease and to produce VSC and to cause bad breath. Peugeot bacterium nucleatum produces fatty acids that are toxic to periodontal tissue bacteria, inhibits the host's immune response and further promotes the growth of spiroheta and the like.

As described above, Porphyromonas gingivalis and Peugeot bacterium nucleatum inhibit the host's immune response, promote the growth of other oral bacteria and contribute to the formation of plaque.

In the extraction in the present invention, in addition to water (hot water), alcohols such as methanol, ethanol or propanol, organic solvents such as ether, chloroform, ethyl acetate, acetone, glycerin, propylene glycol, or appropriately mixed thereof can be used as a solvent. .

According to the present invention, an antimicrobial agent comprising an extract of a wax apple or a water apple is provided. The antimicrobial agent of the present invention exhibits antimicrobial activity specifically to bacteria in the oral cavity, and as a result of use, there is no fear of causing myocerosis.

Moreover, the fruit of a wax apple or a water apple is provided for food, high safety, and the image as a raw material is good. Therefore, the antimicrobial agent containing the extract of a wax apple or a water apple, and the composition for oral cavity containing the same, food-drinks are easy to accept by a consumer.

Although the addition amount to the composition for oral cavity of this invention and food-drinks is not specifically limited, At least 0.005 weight% or more is good. Preferably at least 0.01% by weight is suitable.

EMBODIMENT OF THE INVENTION Hereinafter, although the Example of this invention is given and demonstrated, the scope of the present invention is not limited only to a following example.

Example  1: Preparation of Extract

From the leaves of the wax or water apple, extracts were obtained using various solvents. That is, 20 g (dry weight) of the leaves of the wax or water apple are pulverized, and 200 ml of water, 25% ethanol aqueous solution, 50% ethanol aqueous solution, 75% ethanol aqueous solution, 100% ethanol, 25% methanol aqueous solution, or 50% Extraction was performed in aqueous methanol solution at 70 ° C. for 2 hours.

This was filtered, concentrated with an evaporator, and lyophilized to obtain an extract. The extraction rate was determined from the weight of the extract. Each extraction rate is shown in Tables 1 and 2, respectively.

Extraction Rate of Wax Apple Extract division Extraction rate (%) Wax Apple Leaf Water Extract 19 25% Ethanol Extract of Wax Apple Leaf 24 Wax Apple Leaf 50% Ethanol Extract 29 Wax Apple Leaf 75% Ethanol Extract 23 100% Ethanol Extract of Wax Apple Leaf 20 Wax Apple Leaf 25% Methanol Extract 23 Wax Apple Leaf 50% Methanol Extract 30

Extraction Rate of Water Apple Extract division Extraction rate (%) Water Apple Leaf Water Extract 12 Water Apple Leaf 25% Ethanol Extract 16 Water Apple Leaf 50% Ethanol Extract 17 Water Apple Leaf 75% Ethanol Extract 14 Water Apple Leaf 100% Ethanol Extract 11 Water Apple Leaf 25% Ethanol Extract 11 Water Apple Leaf 50% Methanol Extract 17

Example  2: cultivation of various bacteria

(2-1) Providing Strains

Porphyromonas gingivalis ATCC33277, Peugeot bacterium nucleatum JCM8532, Streptococcus mutants MT8148R, Streptococcus sobrinus 6715, Actinomyces viscous ATCC15987 were used as test strains. For porphyromonas gingivalis, tests were also performed for strains FDC381 and FDA381 other than the above. Peugeot bacterium nucleatum was also tested for strains JCM11023, JCM11024, # 2, and # 20 other than the above.

(2-2) Pre-culture

Porphyromonas gingivalis, Peugeot bacterium nucleatum, Actinomyces viscous, yeast extract (0.3 g), trypticase soy bean (3 g), hemin (0.5 mg), mena Dione (0.05 mg) was added to water and inoculated in a liquid medium prepared in 100 ml, and cultured anaerobicly at 37 ° C. for 24 to 48 hours to prepare a preculture. Streptococcus mutants and Streptococcus sobrinus were inoculated into BHI (Brain Heart Infusion) liquid medium, and cultured at 37 ° C. for 24 hours to prepare a preculture.

Example  3: antimicrobial activity test

Each extract obtained in Example 1 and the extract to be compared were tested for antimicrobial activity. For comparison, water or ethanol extracts of the plating sheep (leaves) and cloves (buds), which are plants of the Myrtaceae sisyygium, were used. The extracts each dissolved 1.6 mg in 1 ml of 2% dimethylsulfoxide (DMSO). These were again serially diluted with an aqueous 2% DMSO solution to prepare a 2-fold dilution series, and 100 μl each was serially dispensed into 96 well plates. To each of these wells 100 μl of diluted bacterial solution was added. In addition, the added diluted bacterial solution was precultured in the liquid medium by the method shown in Example 2, and then, in Porphyromonas gingivalis, Peugeot bacterium nucleatum, and Actinomyces viscous, in the liquid medium of 2 times concentration. The culture solution was diluted 10-fold, and the preculture of Streptococcus mutants and Streptococcus sobrinus was diluted 50-fold in 2-fold concentration of liquid medium. The concentration of each well of the wax and water apple extracts was 800 µg / ml, 400 µg / ml, 200 µg / ml, 100 µg / ml, 50 µg / ml, 25 µg / ml, 12.5 µg / ml, 6.25 µg / ml Ml, 3.13 µg / ml, 1.56 µg / ml, and 0.78 µg / ml. A mixture of this extract and Porphyromonas gingivalis, Peugeot bacterium nucleatum or Actinomyces viscous was incubated at 37 ° C. for 24 to 48 hours under anaerobic conditions. Other mixtures of bacteria and extracts were incubated at 37 ° C. for 24 to 48 hours under aerobic conditions. Minimum Inhibitory Concentration (MIC) of each extract was determined from the turbidity of each well.

(3-1) Antibacterial Activity of Porphyromonas gingivalis and Peugeot bacterium Nucleatum from Wax Apple Extract

The results of the antimicrobial activity test against Porphyromonas gingivalis (ATCC33277) and Peugeot bacterium nucleatum (JCM8532) of the wax apple extract are shown in Table 3 below. The wax apple extract was shown to have antimicrobial activity against Porphyromonas gingivalis, Peugeot bacterium nucleotum.

MIC of Porphyromonas gingivalis, Peugeot bacterium nucleatum of wax apple extract (ppm) division Porphyromonas gingivalis (ATCC33277) Peugeot bacterium nucleatum (JCM8532) Wax Apple Leaf Water Extract 12.5 12.5 25% Ethanol Extract of Wax Apple Leaf 12.5 6.3 Wax Apple Leaf 50% Ethanol Extract 12.5 6.3 Wax Apple Leaf 75% Ethanol Extract 3.1 6.3 100% Ethanol Extract of Wax Apple Leaf 3.1 6.3 Wax Apple Leaf 25% Methanol Extract 12.5 6.3 Wax Apple Leaf 50% Ethanol Extract 6.3 6.3

(3-2) Antibacterial Activity of Water Apple Extract against Porphyromonas gingivalis and Peugeot bacterium Nucleatum

The results of the antimicrobial activity test of Porphyromonas gingivalis (ATCC33277) and Peugeot bacterium nucleatum (JCM8532) of water apple extract are shown in Table 4 below. Water apple extract was shown to have antimicrobial activity against Porphyromonas gingivalis, Peugeot bacterium nucleatum.

MIC of Porphyromonas gingivalis, Peugeot bacterium nucleatum of water apple extract (ppm) division Porphyromonas gingivalis (ATCC33277) Peugeot bacterium nucleatum (JCM8532) Water Apple Leaf Water Extract 25 12.5 Water Apple Leaf 25% Ethanol Extract 25 25 Water Apple Leaf 50% Ethanol Extract 25 25 Water Apple Leaf 75% Ethanol Extract 12.5 50 Water Apple Leaf 100% Ethanol Extract 12.5 50 Water Apple Leaf 25% Methanol Extract 25 25 Water Apple Leaf 50% Ethanol Extract 25 25

(3-3) Antibacterial Activity of Porphyromonas gingivalis (Other Strains) of Wax Apple Extract and Water Apple Extract

The results of the antimicrobial test on porphyromonas gingivalis (FDC381, FDA381) of the wax apple extract and the water apple extract are shown in Table 5 below. The wax apple extract and the water apple extract were found to have antimicrobial activity against strains other than ATCC33277 of Porphyromonas gingivalis.

MIC (ppm) of Porphyromonas gingivalis of Wax Apple Extract and Water Apple Extract division FDA381 FDC381 Wax Apple Leaf Water Extract 12.5 12.5 Water Apple Leaf Water Extract 50 25 Wax Apple Leaf 50% Ethanol Extract 12.5 12.5 Water Apple Leaf 50% Ethanol Extract 50 50

(3-4) Antimicrobial Activity of Waxapple Extract and Waterapple Extract against Peugeot bacterium Nucleatum (Other Strains)

The results of the antibacterial test on the Peugeot bacterium nucleatum (JCM11023, JCM11024, # 2, # 20) of the wax apple extract and the water apple extract are shown in Table 6 below. The wax apple extract and the water apple extract were shown to have antimicrobial activity against strains other than JCM8532 of Peugeot bacterium nucleatum.

MIC (ppm) for Peugeot bacterium nucleatum from Waxapple Extract and Waterapple Extract division JCM11023 JCM11024 #2 # 20 Wax Apple Leaf Water Extract 6.3 12.5 12.5 6.3 Water Apple Leaf Water Extract 25 25 12.5 25 Wax Apple Leaf 50% Ethanol Extract 6.3 12.5 3.1 6.3 Water Apple Leaf 50% Ethanol Extract 25 25 6.3 12.5

(3-5) Antimicrobial Activity of Wax Apple Extract and Water Apple Extract Against Other Bacteria

The results of the antimicrobial test of Streptococcus mutants, Streptococcus sobrinus, and Actinomyces viscous of wax apple extract and water apple extract are shown in Table 7 below. Both wax apple extract and water apple extract did not show antimicrobial activity against these bacteria. The antimicrobial activity of the wax apple extract and the water apple extract was shown to be specific for Porphyromonas gingivalis and Peugeot bacterium nucleotum.

MIC (ppm) for Wax Apple Extract and Water Apple Extract division Streptococcus
Mutance Streptococcus
Sobriners Actinomyces Viscous Wax Apple Leaf Water Extract - - - 25% Ethanol Extract of Wax Apple Leaf - - - Wax Apple Leaf 25% Methanol Extract - - - Water Apple Leaf Water Extract - - - Water Apple Leaf 25% Ethanol Extract - - - Water Apple Leaf 25% Methanol Extract - - -

('-' Means activity is not recognized even at 800ppm.)

(3-6) Antimicrobial Activity of Porphyromonas gingivalis and Peugeot bacterium nucleotum from Comparative Extracts

Water or ethanol extracts of the leaves of the plated sheep, which is a plant of Myrtaceae syyzygium, and the extract of the clove bud which is similarly a plant of the family Syrtygium, Myrtaceae Was used. The results of the antimicrobial activity against Porphyromonas gingivalis (ATCC33277) and Peugeot bacterium nucleatum (JCM8532) are shown in Table 8 below. The antimicrobial activity of the extracts was lower than that of wax apple and water apple, and the antimicrobial activity of Porphyromonas gingivalis was particularly low.

division Porphyromonas gingivalis (ATCC33277) Peugeot bacterium nucleatum (JCM8532) Clove Bud Extract 100 12.5 Clove Bud 50% Ethanol Extract 50 12.5 Clove Bud 100% Ethanol Extract 100 25 50% Ethanol Extract of Plating Lamb Leaves 50 100

Example  4: application

(4-1)

Tablets were prepared according to the following formulation.

D-mannitol 39.6%

Lactose 39.6

Crystalline Cellulose 8.5

Hydroxypropyl Cellulose 4.3

Example  1 of Wax Apple Leaf  Water Extract 8.0

                                 100.0%

This tablet had an antimicrobial effect without losing its properties as a tablet.

(4-2)

The powder was prepared according to the following prescription.

Lactose 62.5%

Potato starch 12.5

Example  1 of Wax Apple Leaf  25% Ethanol Extract 25.0

                                           100.0%

This powder had an antimicrobial effect without losing its properties as a powder.

(4-3)

Toothpaste was prepared according to the following prescription.

Calcium Carbonate 48.0%

Glycerin 20.0

Carboxymethylcellulose 2.0

Sodium Lauryl Sulfate 2.0

Spices 1.0

Saccharin 0.1

Wax Apple Leaf 50% Ethanol Extract of Example 1 3.0

Chlorhexidine 0.01

Water 23.89

                                      100.0%

This toothpaste had an antimicrobial effect without losing its properties as a toothpaste.

(4-4)

Chewing gum was prepared according to the following prescription,

Gum Base 20.0%

Sugar 54.7

Glucose 15.0

Starch syrup 9.3

Spices 0.5

Example  1 of Wax Apple Leaf  75% Ethanol Extract 0.5

                                        100.0%

This chewing gum had an antimicrobial effect without losing its properties as a chewing gum.

(4-5)

Candy was prepared according to the following prescription.

Sugar 49.6%

Starch syrup 33.0

Citric Acid 1.0

Spices 0.2

L-menthol 1.0

Wax Apple Leaf 100% Ethanol Extract of Example 1 0.8

Water 14.4

                                        100.0%

This candy had an antimicrobial effect without losing its properties as a candy.

(4-6)

Chocolate was prepared according to the following prescription.

Kakaobiter 20.0%

Whole milk powder 20.0

Kakaover 17.0

41.75 per minute

Lecithin 0.45

Spices 0.2

Example  1 of Wax Apple Leaf  25% Methanol Extract 0.6

                                        100.0%

This chocolate had an antimicrobial effect without losing its properties as chocolate.

(4-7)

Biscuits were prepared according to the following prescription.

30.7% sugar

Wheat flour 26.8

Flat rate 26.8

Butter 3.2

Egg 10.2

Baking Soda 0.3

Example  1 of Wax Apple Leaf  50% Methanol Extract 2.0

                                        100.0%

This biscuit had an antimicrobial effect without losing its properties as a biscuit.

(4-8)

Gumi jelly was prepared according to the following prescription.

Polydextrose aqueous solution 38.0%

Sorbitol aqueous solution 8.0

Paratinose Aqueous Solution 9.0

Maltose aqueous solution 20.0

Trehalose Aqueous Solution 11.0

Gelatin 10.0

Tartaric Acid 1.0

Example  1 of Water apple leaf Water Extract      3.0

                                 100.0%

This gummi jelly had an antimicrobial effect without losing its properties as a gummi jelly.

(4-9)

The confectionery was prepared according to the following prescription.

72.7% sugar

Lactose 18.9

Sucrose Fatty Acid Ester 0.15

Water Apple Leaf 25% Ethanol Extract 4.0 of Example 1

Water 4.25

                                       100.0%

This fruit had an antimicrobial effect without losing its properties as a fruit.

(4-10)

Ice cream was prepared according to the following prescription.

Egg yolk 11.0%

Sugar 13.5

Milk 37.0

Fresh Cream 37.0

Vanilla Beans 0.5

Example  1 of Water apple leaf  50% Ethanol Extract 1.0

                                       100.0%

This ice cream had an antimicrobial effect without losing its properties as an ice cream.

(4-11)

The beverage was prepared according to the following prescription.

Orange Juicer 30.0%

Iseonghwa Party 15.33

Citric Acid 0.1

Vitamin C 0.04

Spices 0.1

Water Apple Leaf 100% Ethanol Extract of Example 1 0.01

Water 54.42

                                        100.0%

The beverage had an antimicrobial effect without losing its properties.

Claims (6)

An antimicrobial agent showing activity against oral bacteria of Porphyromonas gingivalis or Fusobacterium nucleatum , comprising wax or water apple extracts. The antimicrobial agent according to claim 1, wherein the extract is a leaf extract of a wax apple or a water apple. delete delete The composition for oral cavity containing the antimicrobial agent of Claim 1 or 2. Food and drink containing the antimicrobial agent of Claim 1 or 2.