JP5017531B2 - Mouth cleaner - Google Patents

Description

  The present invention relates to a mouth cleaning agent. More specifically, the present invention relates to a mouth cleanser containing apolactoferrin having antibacterial properties.

  There are three factors that cause caries: microorganisms, teeth, and food. For example, caries are caused by a state in which oral cavity flora is highly caries-induced, a state in which dental cavities are highly sensitive to caries, and a state in which a dietary pattern is highly cariogenic. The mechanism of caries development is that the caries fungus Streptococcus mutans produces glucosyltransferase (GTase), a plaque-forming enzyme, and is a glucan that is an insoluble and sticky polysaccharide using sucrose as a substrate. Is synthesized. Synthesized glucan adheres to the tooth surface together with caries bacteria to form plaques, and lactic acid bacteria (Lactococcus lactis subsp. Lactis) produce lactic acid in the plaques to demineralize enamel.

  It has been reported that lactoferrin has a pathogen adherence-inhibiting action as a substance having a caries-suppressing effect, that is, a caries-preventing effect (Patent Document 1). Furthermore, Patent Document 2 describes that lactoferrin is combined with other substances (lactoperoxidase, tea polyphenol, fluorine) to increase the caries inhibitory effect.

  Lactoferrin is present in body fluids of many mammals, such as milk. In particular, it is known that colostrum of breast milk contains 5 to 10 g / L and occupies 30% to 70% of the total protein contained. Lactoferrin is an important protein for maintaining and developing the health of infants, and has recently been found to have antibacterial and antibacterial effects, and is used in various fields in addition to the food industry.

  Lactoferrin is an iron-binding glycoprotein having a molecular weight of about 80,000, in which two irons are bound in one molecule. Lactoferrin releases iron under acidic conditions such as pH 2 to become apolactoferrin. Regarding the bacteriostatic (antibacterial) action of lactoferrin, it was considered as follows. Due to the chelating action of apolactoferrin, the iron necessary for the growth of microorganisms is deprived and its growth is restricted. For this reason, microorganisms that strongly require iron during growth receive the bacteriostatic (antibacterial) action of lactoferrin. Such a bacteriostatic (antibacterial) action of lactoferrin has been considered particularly in the intestinal environment. Thus, the chelating action of apolactoferrin has attracted attention regarding the bacteriostatic (antibacterial) action of lactoferrin. However, there are many parts that are not yet known about the properties of apolactoferrin itself.

Patent Document 3 describes that antibacterial properties against skin resident pathogenic bacteria and cariogenic bacteria tend to increase by hydrolyzing lactoferrin with an acid. More specifically, bovine lactoferrin is adjusted to pH 3 or lower with citric acid and then treated at high temperature (95 ° C.) for 5 minutes to 3 hours to obtain various iron binding ability products. It is described that products with lower potency had higher antibacterial effects against skin resident pathogens and cariogenic bacteria
JP-A-3-220130 Japanese Patent Laid-Open No. 9-107917 JP-A-6-279310

  An object of the present invention is to provide an oral cleanser having high antibacterial activity against caries bacteria (cariogenic bacteria).

The present invention provides a mouth cleanser containing apolactoferrin, wherein the mouth cleanser comprises:
The iron binding degree of the apolactoferrin is 5% or less, and when an aqueous solution containing the apolactoferrin at a concentration of 1 w / v% is prepared, the total cation concentration in the aqueous solution is 5 mmol / L or less.

  ADVANTAGE OF THE INVENTION According to this invention, the mouth cleaning agent with high antimicrobial property with respect to a caries fungus (cariogenic bacteria) is provided.

(Apolactoferrin)
The mouth cleanser of the present invention contains apolactoferrin. Apolactoferrin is a glycoprotein molecule from which iron bound in the lactoferrin molecule is released. The apolactoferrin used in the present invention is not particularly limited as long as it has the following characteristics.

  The apolactoferrin in the present invention has an iron binding degree in the molecule of 5% or less, preferably 4% or less, more preferably 3% or less. Here, the degree of iron binding refers to the ratio of the number of moles of iron to the number of moles of apolactoferrin. The degree of iron binding can be determined by measuring the absorbance of apolactoferrin by spectroscopic analysis or by directly measuring the amount of iron in apolactoferrin by atomic absorption analysis or ICP spectroscopy. In the present invention, the degree of iron binding refers to a value obtained by dissolving apolactoferrin powder in pure water to make a 1 w / v% solution, and measuring this at an absorbance of 470 nm.

  In the case of preparing an aqueous solution containing apolactoferrin at a concentration of 1 w / v%, the total cation concentration in the aqueous solution is 5 mmol / L or less. The total cation concentration is determined by dissolving apolactoferrin powder in 0.1N hydrochloric acid to prepare a 0.1 w / v% solution, and measuring the amount of each cation by atomic absorption photometry. And add them together. The total cation concentration may correspond to a salt (ion) contained as an impurity in the apolactoferrin powder. This is because not the ions bound to lactoferrin but only the salt mixed in the powder can be dissolved by the above 0.1N hydrochloric acid. The total cation concentration is preferably 3.0 mmol / L or less, and more preferably 1.0 mmol / L or less.

  Apolactoferrin can be usually produced by adjusting the pH of an aqueous solution containing lactoferrin to the acidic side to dissociate divalent iron ions of the lactoferrin molecule.

  Lactoferrin, which is a raw material for apolactoferrin, is separated and purified from mammalian secretions such as milk (eg, milk) or processed milk products such as skim milk and whey (eg, whey) (eg, adsorbed on a cation exchange resin). Then, it may be obtained by utilizing a method of desorption with a high-concentration salt solution, a separation method by electrophoresis, a separation method by affinity chromatography, or the like. Furthermore, it may be produced by various cells (including microorganisms, plant cells, animal cells, insect cells, etc.) obtained by genetic recombination, plants, animals and the like. Lactoferrin may be commercially available as pharmaceuticals, reagents, and the like. Lactoferrin is preferably derived from natural products. Preferably, it is derived from whey. Whey obtained as a by-product generated when producing a dairy product (for example, cheese, casein, etc.) from cow milk or skim milk can be suitably used as a source of lactoferrin.

  Apolactoferrin can be preferably produced, for example, by adding an acid to the solution when ultrafiltration of the lactoferrin-containing solution to dissociate iron ions bound to lactoferrin. Examples of the acid that can be used here include citric acid, hydrochloric acid, phosphoric acid, malic acid, and acetic acid (0.4 M or more), and citric acid is preferable. Alternatively, for example, apolactoferrin is partitioned by these membranes in which bipolar membranes and cation exchange membranes, which are composite ion exchange membranes having a structure in which a cation exchange membrane and an anion exchange membrane are laminated, are alternately arranged. It can be suitably manufactured also by using an electrodialyzer having an acid chamber and a base chamber. In this case, hydrochloric acid produced during the manufacturing process in the electrodialyzer is used as the acid.

  In the production of apolactoferrin in the present invention, the adjusted acidic pH is preferably 0.5 to 3.0, more preferably 1.5 to 2.5. When the pH is close to neutral (for example, 5.5), the antibacterial properties of the obtained apolactoferrin may be weakened. As a pH adjuster of an aqueous solution containing lactoferrin, not only the acid but also phthalic acid, glycine and the like can be used. These pH adjusting agents are added to an aqueous solution containing lactoferrin in an amount suitable for adjusting the pH to the above value.

  The temperature at which the pH of the aqueous solution containing lactoferrin is adjusted to the acidic side is preferably not high considering protein denaturation. Usually, it is 5 to 60 ° C, more preferably 15 to 35 ° C, and even more preferably room temperature.

  Specific production of apolactoferrin in the present invention will be described in detail in the following preparation examples, but the production method of apolactoferrin is not limited thereto. The apolactoferrin in the present invention can also be obtained by modifying a commercially available apolactoferrin so as to have the above iron binding degree and total cation concentration.

  In the production of apolactoferrin, apolactoferrin can usually be obtained in the form of an aqueous solution. When the mouth cleanser of the present invention is prepared using apolactoferrin, it may be used in the form of an aqueous solution or in a powdered form after removing the solvent.

(Oral cleanser)
The content of apolactoferrin in the mouth cleanser of the present invention is not particularly limited because it varies depending on the dosage form. Preferably they are 0.01 mass%-20 mass%, More preferably, they are 0.1 mass%-10 mass%, More preferably, they are 1 mass%-5 mass%.

  The mouth cleanser of the present invention is intended for the prevention and treatment of oral diseases such as bad breath, caries and periodontal disease. The mouth cleanser of the present invention can take any form that is applicable to the oral cavity. Examples of the dosage form of the external preparation of the present invention include lotions, emulsions, sprays, gels, powders, granules, tablets and the like. The mouth cleanser of the present invention can be used as pharmaceuticals, quasi drugs, toiletries, oral preparations, and the like. More specifically, dentifrice, mouthwash, spray, troche and the like can be mentioned.

  In addition to the apolactoferrin described above, the mouth cleanser of the present invention may contain appropriate components depending on the purpose, the type of composition, and the like.

  For example, a fragrance or the like as a refreshing agent such as a bactericidal agent, an anti-inflammatory agent, a hemostatic agent, a tissue activator, etc., used for prevention or treatment of normal oral diseases can be blended.

  In addition, solvents, thickeners, surfactants, abrasives, sweeteners, preservatives, pigments, and the like can be blended.

  For example, examples of the solvent include ethyl alcohol, propylene glycol, polyethylene glycol, maltite, and lactitol. Examples of the thickener include carboxyvinyl polymer and / or a salt thereof, sodium carboxymethyl cellulose, hydroxyethyl cellulose, carrageenan, montmorillonite, gelatin and the like.

  Examples of the surfactant include an anionic surfactant, a nonionic surfactant, an amphoteric surfactant, and a cationic surfactant. More specifically, sodium lauryl sulfate, sucrose fatty acid ester, lactose fatty acid ester, lactitol fatty acid ester, maltitol fatty acid ester, stearic acid monoglyceride, polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan monostearate, polyoxy Ethylene (10, 20, 40, 60, 80, 100 mol) hydrogenated castor oil, polyoxyethylene alkyl ether, polyoxyethylene polyoxypropylene monolauryl ester and other polyethylene oxides and fatty acids, alkylol amide, 2-alkyl-N -Carboxymethyl-N-hydroxyethylimidazolinium betaine and the like.

  Examples of the abrasive include silica-based abrasives such as precipitated silica and zirconosilicate, calcium phosphate, aluminum hydroxide, and aluminum oxide.

  Examples of the sweetener include saccharin sodium, stevioside, stevia extract, paramethoxycinnamic aldehyde, neohesperidyl dihydrochalcone, and perilartin. Examples of the preservative include paraoxybenzoic acid ester and sodium benzoate. Examples of the colorant include blue No. 1, yellow No. 4, and titanium dioxide.

  In addition, the compounding quantity of these components can be made into a normal quantity in the range which does not prevent the mouth cleaning effect in this invention.

  The mouth cleanser of the present invention contains components usually used by those skilled in the art as described above, if necessary, and can be prepared according to methods commonly used by those skilled in the art. Apolactoferrin can be added to, blended with, or contained in the mouthwash by procedures commonly used by those skilled in the art. For example, it can be added to, blended with, or contained in the mouth cleansing agent by dissolving in advance in a solvent such as water or alcohol and then mixing with other compounding ingredients, or by stirring and mixing with other compounding ingredients as a powder. .

  The mouth cleanser of the present invention is in the form as described above, and is administered or applied to an individual according to the means normally used for administration or application in that form (for example, spray, gargle, dissolution in the oral cavity). Can be done.

  EXAMPLES Hereinafter, although an Example demonstrates this invention more concretely, this invention is not limited to these illustrations.

(Preparation Example 1: Production of apolactoferrin by various acid treatments)
LOV (hollow fiber module: membrane inner diameter 0.8 mm, effective membrane area 41 m ² , membrane material: polyacrylonitrile, nominal, manufactured by Microza UF Lab Test Machine (LX-22001; Asahi Kasei Chemicals Corporation)) Using an ultrafiltration apparatus incorporating a molecular weight cut off: 50,000), apolactoferrin was produced as follows.

  10 kg of 50 mg / mL lactoferrin (manufactured by Fontera; iron binding degree is about 20%) was used. In the manufacturing process of apolactoferrin, lactoferrin was treated with any of the following acids: 0.1 M citric acid, 0.1 M lactic acid, 0.1 M hydrochloric acid, 0.1 M malic acid, 0.1 M acetic acid, or 0.4 M Acetic acid. First, the lactoferrin solution was put into a supply tank of the apparatus, circulated for 10 minutes, and then circulated in the reverse direction for 5 seconds to concentrate the solution. At this time, the pressure and circulating fluid flow rate at the inlet and outlet of the UF membrane were set to 0.12 Mpa, 0.08 Mpa, and 15 L / min, respectively. This operation was repeated until the non-permeated concentrate was halved (this was defined as one round). Next, the citric acid solution was put into the tank instead of the lactoferrin solution, and the same operation as above was performed for two rounds. Subsequently, 8 MΩ · cm or more of pure water was put into the tank, and the above operation was performed for 5 rounds to remove the acid remaining in the non-permeated concentrate. The temperature of the circulating liquid was in the range of 10 to 28 ° C. throughout the production process, and the pH was 2 to 3.

  A 40 kg concentrated solution was obtained by the above production process. The concentrated solution was then lyophilized to obtain 9.5 g of white powder.

The powder obtained by each acid treatment was apolactoferrin and the purity of apolactoferrin was measured using BIOXYTECH (registered trademark) Lacto f EIA ^™ (OXIS International Inc., Oregon, USA) after dissolving the powder in pure water. Determined by performing antibody quantification. The purity of apolactoferrin was as follows: 93% for 0.1M citric acid, 94% for 0.1M lactic acid, 96% for 0.1M hydrochloric acid, 91% for 0.1M malic acid,. 91% for 1M acetic acid and 91% for 0.4M acetic acid.

  Furthermore, the iron binding degree of the powder obtained by each acid treatment is dissolved in pure water so as to have a concentration of 1 w / v%, and then the amount of iron bound to apolactoferrin is measured at an absorbance of 470 nm. Determined by measuring. Table 1 shows the degree of iron binding of the obtained apolactoferrin. Here, the degree of iron binding was calculated by the degree of iron binding (%) = (number of moles of iron in a 1 w / v% solution / 1 number of moles of apolactoferrin in a 1 w / v% solution) × 100.

(Preparation Example 2: Production of apolactoferrin by various citrate treatment times)
Except for increasing or decreasing the number of citric acid treatments, an apolactoferrin powder was produced according to the procedure described in Preparation Example 1 above, and the degree of iron binding was measured. Table 2 shows the degree of iron binding of the obtained apolactoferrin.

(Example 1: Apolactoferrin antibacterial test using E. coli)
The antibacterial properties of the apolactoferrin powders produced in the above Preparation Example 1 and treated with various acids were examined. As a control, lactoferrin powder (manufactured by Fontera) was used.

E. coli (NBRC 3972) purchased from the National Institute of Biomedical Resources (NBRC), Biotechnology Headquarters, National Institute for Product Evaluation Technology, subcultured in 5 mL of SCD bouillon (Nissui Pharmaceutical Co., Ltd.) Liquid). 50 μL of this stored Escherichia coli solution was inoculated into 5 mL of SCD broth and cultured in a shaking water bath at 30 ° C. for 16 hours (preculture). It was diluted pre-cultured bacterial solution with sterile water to prepare a 10-fold serial dilutions of up to 10 ⁷ times. In each well of a 96-well flat-bottom microplate (BD Falcon), 50 μL of a series obtained by dissolving apolactoferrin or lactoferrin powder in sterilized water and diluting multiple times, 100 μL of SCD broth having a double concentration, 50 μL of 10 ⁵ diluted bacterial solution was added. The microplate was cultured at 35 ° C. for 24 hours. After culturing, the microplate was measured for turbidity (absorbance) at a wavelength of 630 nm with a microplate reader (Multiscan JX: Thermo Labsystems) to confirm the antibacterial action.

  FIG. 1 is a graph showing the absorbance in an E. coli culture solution when apolactoferrin or lactoferrin obtained by each acid treatment is added at various concentrations. The horizontal axis indicates the apolactoferrin or lactoferrin concentration (%) in the culture solution. The vertical axis represents the absorbance, and the lower the absorbance, the smaller the viable cell count, that is, the higher the antibacterial activity. As is clear from FIG. 1, lactoferrin showed little antibacterial action, whereas apolactoferrin showed antibacterial action as the concentration of addition increased. In particular, the antibacterial effect of apolactoferrin obtained by the citric acid treatment was most excellent.

(Example 2: Influence of the degree of iron binding in an apolactoferrin antibacterial test using Escherichia coli)
The antibacterial activity against Escherichia coli was examined in the same manner as in Example 1, using the apolactoferrin obtained in Preparation Example 2, a commercially available lactoferrin (manufactured by Fontera), and a commercially available apolactoferrin (manufactured by Tatsua Dairy). Here, the degree of iron binding of commercially available lactoferrin was 19.8%, and the degree of iron binding of commercially available apolactoferrin was 4.39%.

  FIG. 2 is a graph showing the absorbance in an E. coli culture solution when apolactoferrin having various iron binding degrees is added at various concentrations. The horizontal axis indicates the concentration of apolactoferrin added (%), and the vertical axis indicates the absorbance. The lower the absorbance, the lower the viable cell count, that is, the higher the antibacterial activity. From FIG. 2, lactoferrin having an iron binding degree of 19.8% did not decrease in absorbance and no antibacterial action was observed. In the case of apolactoferrin having an iron binding degree of 5% or less, the absorbance decreased with an increase in the addition amount, and an excellent antibacterial action was observed. Furthermore, apolactoferrin having an iron binding degree of about 4% or less (iron binding degree: 4.09%, iron binding degree: 2.95%, iron binding degree: 2.2%, iron binding degree: 1.66%, and iron binding degree) 1.04%), sufficient antibacterial activity was also observed in an aqueous solution containing apolactoferrin at a concentration of 5 w / v%.

(Example 3: Effect of salt in an apolactoferrin antibacterial test using Escherichia coli)
In the antibacterial test of Escherichia coli of Example 1 above, the influence of salt was examined using citrate-treated apolactoferrin. Antibacterial activity was examined according to the same procedure as in Example 1 except that sodium chloride was added to a 1 w / v% citric acid-treated apolactoferrin solution so as to have various ion concentrations.

  FIG. 3 is a graph showing the absorbance in an E. coli culture solution containing various concentrations of sodium chloride and 2.0 w / v% apolactoferrin. The horizontal axis indicates sodium chloride concentration (mmol / L), and the vertical axis indicates absorbance. The lower the absorbance, the lower the viable cell count, that is, the higher the antibacterial activity. In FIG. 3, the antibacterial effect decreased as the salt addition concentration increased. Therefore, it is understood that the amount of salt that can be mixed as impurities in the apolactoferrin powder affects the antibacterial properties. Furthermore, it has become clear that the amount of such salt is desirably 5 mmol / L or less based on the total ions when apolactoferrin is used as a 1% solution.

  Therefore, the total cation concentration of apolactoferrin with an iron binding degree of 2.95% obtained in Preparation Example 2 was determined. By adding 0.1N hydrochloric acid to the lyophilized powder of apolactoferrin, preparing a 0.1 w / v% apolactoferrin solution and measuring for Na, K, Ca, Mg, and Cu by atomic absorption spectrophotometry, these The concentration of each cation was determined and the total was taken as the total cation concentration. For comparison, the total cation concentration of commercial lactoferrin and apolactoferrin was also determined. The results are shown in Table 3.

  The apolactoferrin with an iron binding degree of 2.95% obtained in Preparation Example 2 had a total cation concentration of 5 mmol / L or less. In Example 2, with the commercially available apolactoferrin (manufactured by Tatsua Dairy), the absorbance decreased to a certain level or more even though the iron binding degree was 5% or less. As shown in this example, this apolactoferrin has a total cation concentration exceeding 5 mmol / L. Therefore, it was considered that both the iron binding degree and the total cation concentration of the apolactoferrin powder affect the antibacterial properties.

(Example 4: Antibacterial test of apolactoferrin having predetermined iron binding degree and total cation concentration)
Considering the results of Example 3 above, the antibacterial properties of apolactoferrin having an iron binding degree of 5% or less and a total cation concentration of 5 mmol / L or less were further examined. The following three types of apolactoferrin were used: Sample A (commercial apolactoferrin: degree of iron binding 4.39%, total cation concentration 14.7 mmol / L); sample B (sample A was prepared according to the method of Preparation Example 1) Retreated at a temperature of 25 ± 1 ° C. and a pH of 2.6: iron binding degree 3.89%, total cation concentration 4.5 mmol / L; and sample C (apolactoferrin obtained in Preparation Example 2 above): Iron binding degree 4.56%, total cation concentration 4.4 mmol / L).

  About antibacterial property, it carried out in the same procedure as Example 1. The final concentration of apolactoferrin was 20 mg / mL.

  FIG. 4 shows a graph showing the absorbance in the culture solution of E. coli when samples A, B, and C are added. The vertical axis represents the absorbance, and the lower the absorbance, the smaller the viable cell count, that is, the higher the antibacterial activity. In the sample A in which the degree of iron binding was 5% or less but the total cation concentration was higher than 5 mmol / L, the absorbance was not so different as compared to the culture solution to which no apolactoferrin was added. On the other hand, Samples B and C, which have an iron binding degree of 5% or less and a total cation concentration of 5 mmol / L, have a low absorbance compared to the culture solution to which no apolactoferrin is added. Showed antibacterial properties.

(Example 5: Effect of apolactoferrin on cultured caries fungus)
Regarding the apolactoferrin having an iron binding degree of 2.95% obtained in Preparation Example 2 above, two types of Streptococcus mutans (obtained from Air Brown Co., Ltd .: 25175 strain (S. mutans MT) and 35668 strain (S .mutans XC)), its growth inhibitory effect was examined. As a control, lactoferrin (iron binding degree: about 20%; manufactured by Fontera) was used.

  5 ml of Todd Hewitt Broth (TH medium: manufactured by Kanto Chemical Co., Ltd.) placed in a test tube was inoculated with mutans bacteria, and left to stand at 37 ° C. for 24 hours in an incubator.

Apolactoferrin powder and lactoferrin powder were dissolved in water to give an 8 w / v% aqueous solution. Next, the 8 w / v% aqueous solution was diluted to prepare 4 w / v%, 2 w / v%, 1 w / v%, and 0.5 w / v% aqueous solutions. 50 μL of each of these aqueous solutions, 50 μL of cultured mutans (10 ⁴ diluted) bacterial solution, and 100 μL of TH medium were placed in a microplate and cultured at 37 ° C. for 24 hours. After incubation, the absorbance of the microplate was measured at 595 nm to determine the bacterial concentration of 25175 strain, and the absorbance was measured at 630 nm to determine the bacterial concentration of 35668 strain.

  FIG. 5 is a graph showing the absorbance in each mutans culture medium when various concentrations of apolactoferrin (Apo-Lfn) or lactoferrin (Lfn) are added (A: mutans 25175 strain, B: mu Tans bacteria 35668). The horizontal axis indicates the concentration of apolactoferrin or lactoferrin added (%), and the vertical axis indicates the absorbance. The lower the absorbance, the lower the viable cell count, that is, the higher the antibacterial activity. As shown in FIG. 5, both A: S. mutans strain 25175 and B: S. mutans strain 35668 showed an antibacterial action as the absorbance decreased with increasing amount of apolactoferrin. From this, it was found that apolactoferrin exhibits a strong mutans bacterium inhibitory action.

(Example 6: Mutans bacteria count level test in human oral cavity)
Twenty healthy volunteers were recruited, a medical examination was conducted by a doctor, and only those who met the test criteria of this study were entered. Grouped and treated as follows:
Group A (7 persons): Day 1-untreated, Day 2-Apolactoferrin (ApoLfn) gargle, Day 3-Water gargle;
Group B (7 people): Day 1-water gargle, Day 2-untreated, Day 3-ApoLfn gargle; and Group C (6 people): Day 1-ApoLfn gargle, Day 2-water gargle, Day 3-no treatment.

  Here, “no treatment” means that no mouth washing is performed, and “water gargle” means that the mouth is rinsed by gargle once for 5 seconds in three portions with 150 mL of tap water. “Apolactoferrin (ApoLfn) gargle” means that 150 mL of 2% avactoferrin aqueous solution produced in Preparation Example 1 is divided into 3 portions, and the oral cavity is rinsed by gargle once in 5 seconds. To do.

  After 21:00 on the day before the test, they did not eat and brushed their teeth. Drinking and smoking were also prohibited after 3 o'clock on the test day. In addition, violent exercise was banned from the previous day. Each group member was treated as described at 7 am. At 9 o'clock, in a relaxed state, the chewing gum contained in the kit "Oral Tester (registered trademark)" (Tokuyama Dental Co., Ltd.) was chewed for 5 minutes. Saliva was put into a container, 0.5 mL was sucked with a syringe, and the syringe filter included in the kit was attached and filtered. Next, the entire amount of the treatment liquid 1 contained in the kit was sucked and filtered again. Next, the entire amount of the treatment solution 2 contained in the kit was sucked, filtered, and allowed to stand for 2 minutes. Next, the entire amount of the treatment liquid 3 contained in the kit was sucked and discharged to the treatment liquid tray contained in the kit. 0.1 mL of the collected liquid (antigen) was sucked using a mutans dropper and added to the sample window of the chromatographic device. 10 to 20 minutes after addition, using the determination chart, “low (score 0)”, “slightly high (score 1)”, “high (score 2)”, and “very high (score 3)” It was judged. The same test was conducted on the second and third days. Therefore, the same subject has received three treatments. The results were summarized for each treatment and expressed as the mean ± SD of 20 subjects (no dropouts).

Figure 6 is a graph showing the mutans count score in the oral cavity in water gargling and apo lactoferrin gargle. In the figure, the “Control group” is a group that summarizes the results of no treatment, the “water gargle group” is a group that summarizes the results of the gargle treatment, and the “ApoLfn gargle group” is an apolactoferrin gargle treatment This is a group that summarizes the results. As shown in FIG. 6, in the apolactoferrin gargle group, the number of mutans bacteria in the oral cavity was significantly reduced compared to the Control group (p <0.01). Therefore, mutans bacteria in the oral cavity were suppressed by gargle with apolactoferrin.

  The mouth cleanser of the present invention has high antibacterial properties against caries bacteria (cariogenic bacteria). Also, it is highly safe and cannot cause side effects. Therefore, the mouth cleanser of the present invention is useful for preventing tooth decay, bad breath and the like.

It is a graph which shows the light absorbency in colon_bacillus | E._coli culture liquid at the time of adding the apolactoferrin or lactoferrin obtained by each acid treatment at various concentrations. Is a graph showing the absorbance at E. coli culture when adding apo lactoferrin iron binding of the species people at various concentrations. It is a graph which shows the light absorbency in a E. coli culture solution at the time of adding apolactoferrin with various concentration sodium chloride. It is a graph which shows the light absorbency in the culture solution of colon_bacillus | E._coli at the time of adding the samples A, B, and C which are apolactoferrins with various iron binding degrees and various total cation concentrations. It is a graph which shows the light absorbency in each mutans culture liquid at the time of adding various concentrations of apolactoferrin or lactoferrin It is a graph which shows the mutans bacteria count score in the oral cavity in a water gargle and an apolactoferrin gargle.

Claims (3)

An oral cleanser containing apolactoferrin,
When an aqueous solution containing the apolactoferrin having an iron binding degree of 4.09 % or less and containing the apolactoferrin at a concentration of 1 w / v%, the total cation concentration in the aqueous solution is 5 mmol / L or less. is there,
Mouth cleaner. The oral cleaner of Claim 1 containing 0.1 mass%-10 mass% of the apolactoferrin. The oral cleanser according to claim 1 or 2, wherein the apolactoferrin is an apolactoferrin obtained by ultrafiltration of a lactoferrin-containing liquid adjusted to be acidic by adding an acid.

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