An anti-sugar composition containing hydrolyzed wheat protein and use thereof

Abstract

The application belongs to the technical field of oral care, and discloses an anti-sugar composition containing hydrolyzed wheat protein and application thereof.The anti-sugar composition containing hydrolyzed wheat protein comprises the following components in mass fraction: 0.001-0.1 parts of glucose oxidase, 0.001-0.1 parts of dextranase, 0.001-10 parts of hydrolyzed wheat protein, 0.01-0.5 parts of arginine, 0.01-0.05 parts of fluoride and 0.001-0.1 parts of probiotics.The anti-sugar composition is prepared by scientific proportioning and synergistic effect, and the prepared oral care product can effectively decompose dental plaque, reduce dental plaque, inhibit the generation of bacteria, inhibit tooth demineralization and promote tooth remineralization, reduce the toxic and side effects of fluoride and maintain oral health.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of oral care, in particular to an anti-sugar composition containing hydrolyzed wheat protein and application thereof. BACKGROUND

[0002] With the improvement of living standards, the degree of fine processing of sugar-containing food is also increasing, and the intake of sugar-containing food and sugar-containing beverages is on the rise, among which children are most affected. According to the epidemiological survey of oral health, the permanent tooth caries rate of 12-year-old children is 34.5%, which is 7.8 percentage points higher than ten years ago. The deciduous tooth caries rate of 5-year-old children is 70.9%, which shows that the oral health status of children is not optimistic. Children are a high-risk group of dental caries, and the occurrence of dental caries is closely related to sugar intake and oral hygiene. In addition, some acidic foods and acidic beverages are also the main factors for the destruction of the oral environment balance of children and the acid erosion of dental enamel. Due to the lack of self-consciousness and the necessary understanding of oral care, children cannot actively develop good brushing habits, and often eat snacks such as candies and biscuits with high sugar content, and parents do not guide them in time, so many children have oral diseases such as acid erosion and pain of teeth at an early age, which directly affects the learning, life and overall health of children.

[0003] Food residues in children's oral cavity contain a lot of sugar substances, which can become nutrients for oral bacteria. Common oral pathogenic bacteria in children's oral cavity mainly include Streptococcus mutans, Porphyromonas gingivalis, Fusobacterium nucleatum, Lactobacillus acidophilus, etc. These oral bacteria can produce a large amount of organic acid in the process of metabolizing sugar, which can penetrate the surface of dental enamel and cause the loss of tooth mineral (hydroxyapatite) and demineralization, resulting in the phenomenon of acid erosion of teeth. Further, the reproduction of oral bacteria can form a layer of adherent film-dental plaque on the surface of teeth, so that the acid produced by bacteria can adhere to the surface of teeth for a long time, so that hydroxyapatite is dissolved by acid to generate hydrogen phosphate ion and calcium ion, which is washed away by saliva, eventually eroding teeth, accelerating the demineralization of teeth, and causing a vicious cycle. Therefore, the anti-sugar effect of children's toothpaste is particularly important.

[0004] Fluoride can inhibit bacterial acid production, inhibit enamel demineralization and promote remineralization, and has been widely used in the oral care industry. However, long-term use also needs to consider its safety to avoid complications such as dental fluorosis. Currently, the toothpaste with anti-sugar effect on the market only relies on sodium fluoride to achieve the effect of preventing dental caries, and the anti-sugar path is single. There is no in-depth research on the anti-sugar ingredients and mechanism of action, and it is difficult to effectively balance the removal of dental plaque biofilm and the removal of dental plaque. In addition, in addition to fluoride-containing toothpaste, there is no children's toothpaste containing ingredients for repairing and remineralizing dental enamel. Therefore, developing a new type of anti-sugar (resisting the erosion of dental enamel caused by glycolysis) oral care product not only meets the needs of consumers, but also has broad market application prospects. SUMMARY

[0005] The present application aims to overcome the shortcomings of the prior art and provide an anti-sugar composition containing hydrolyzed wheat protein and its application.

[0006] To achieve the above-mentioned purpose, the technical solution adopted by the present application is as follows:

[0007] In a first aspect, the present application provides an anti-sugar composition containing hydrolyzed wheat protein, which comprises the following components in mass fraction: hydrolyzed wheat protein 0.001-10 parts, glucose oxidase 0.001-0.1 parts, dextranase 0.001-0.1 parts, arginine 0.01-0.5 parts, fluoride 0.01-0.05 parts, and probiotics 0.001-0.1 parts.

[0008] The anti-sugar composition of the present application adopts a scientific ratio. Glucose oxidase can decompose glucose, reduce the generation of bacteria, and inhibit the formation of dental plaque. Dextranase can effectively decompose dental plaque biofilm and reduce the production of dental plaque. Probiotics can inhibit the growth and reproduction of oral pathogenic bacteria such as Streptococcus mutans, Porphyromonas gingivalis, and Fusobacterium nucleatum, and maintain the balance of oral microecology. Hydrolyzed wheat protein can combine with calcium ions, adsorb on the surface of dental enamel, and promote the repair and remineralization process of teeth. Arginine can neutralize the acid produced by bacteria in the oral cavity, inhibit tooth demineralization, and promote tooth remineralization. Fluoride can enhance the hardness of teeth, promote tooth remineralization, and inhibit the growth of acid-producing bacteria. The various components have a synergistic effect, and the stability of the anti-sugar children's toothpaste prepared therefrom is significantly improved. The bioactive ingredients have high activity, thereby ensuring the stability of the various active substances within the shelf life of the product. Repeated use over a period of several days can effectively resist sugar, including helping to inhibit oral bacteria, remove dental plaque biofilm, inhibit dental plaque formation, resist bacterial glycolysis and acid production, and resist tooth demineralization caused by glycolysis and acid production, promote tooth remineralization, etc.

[0009] As a preferred embodiment of the anti-sugar composition of the present application, the ratio of the two added amounts of the dextranase and the glucose oxidase is 0.001:1 to 1:1. Preferably, the ratio is 0.01:1 to 1:1.

[0010] As a preferred embodiment of the anti-sugar composition of the present application, the ratio of the two added amounts of the dextranase or the glucose oxidase and the hydrolyzed wheat protein or the fluoride is 0.001:1 to 1:1. Preferably, the ratio is 0.01:1 to 1:1.

[0011] In a second aspect, the present application applies the anti-sugar composition in oral care.

[0012] In a third aspect, the present application provides an anti-sugar oral care product comprising 0.001% to 5% of the anti-sugar composition by mass percentage. Preferably, the anti-sugar oral care product comprises 0.01% to 5% of the anti-sugar composition; more preferably, the anti-sugar oral care product comprises 0.01% to 3% of the anti-sugar composition.

[0013] As a preferred embodiment of the anti-sugar oral care product of the present application, the anti-sugar oral care product is an anti-sugar children's toothpaste.

[0014] As a preferred embodiment of the anti-sugar oral care product of the present application, the anti-sugar children's toothpaste further comprises the following components by mass percentage: a humectant 25% to 28%, a natural antibacterial agent 0.1% to 1%, a thickening agent 0.2% to 1%, a soft abrasive 10% to 25%, xylitol 0.01% to 20%, a calcium-containing compound 0.001% to 5%, a mild surfactant 0.01% to 2%, a fragrance 0.5% to 1%, and the balance being water.

[0015] As a preferred embodiment of the anti-sugar oral care product of the present application, the natural antibacterial agent is at least one of benzyl alcohol, caprylyl glycol, hexylene glycol, and p-hydroxyacetophenone.

[0016] As a preferred embodiment of the anti-sugar oral care product of the present application, the calcium-containing compound is at least one of soluble calcium or slightly soluble calcium.

[0017] As a preferred embodiment of the anti-sugar oral care product of the present application, the mild surfactant is at least one of sodium lauroyl methyl amino acid, sodium lauroyl glutamate, cocamidopropyl betaine, sodium N-methyl cocoyl taurate, poloxamer, and alkyl polyglycoside.

[0018] Compared with the prior art, the present application has the following beneficial effects:

[0019] The anti-sugar composition of the present application adopts scientific proportioning, and each component synergistically enhances the effect. The dextranase and glucose oxidase decompose dental plaque biofilm by dissolving the extracellular polysaccharide of the dental plaque biofilm, remove dental plaque and inhibit the formation of dental plaque. The anti-sugar composition can inhibit or kill harmful bacteria such as Escherichia coli, Staphylococcus aureus, Streptococcus mutans, Porphyromonas gingivalis, Fusobacterium nucleatum, etc., and the bacteriostatic rate is above 97.5%, thereby preventing or assisting in treating oral diseases caused by oral pathogenic bacteria. The anti-sugar composition has the multiple-path anti-sugar effects of inhibiting the generation of oral pathogenic bacteria, resisting the acid production of dental plaque, inhibiting the formation of dental plaque, removing dental plaque and dental calculus, resisting the acid erosion of dental enamel caused by the glycolysis of oral bacteria, and promoting the remineralization of teeth. The prepared toothpaste can maintain a high enzyme activity formula, and the enzyme activity is as high as 73% to 85% when stored at 45°C for 2 months. BRIEF DESCRIPTION OF DRAWINGS

[0020] Figure 1 Statistical diagram for the effect of removing dental calculus of the anti-sugar children toothpaste;

[0021] Figure 2 Graphical representation of the in-vitro evaluation experiment of inhibiting the glycolysis of dental plaque. DETAILED DESCRIPTION

[0022] For the purpose of better illustrating the object, technical scheme and advantages of the present application, the present application will be further described in conjunction with specific embodiments. Those skilled in the art should understand that the specific embodiments described herein are only used to explain the present application, and are not used to limit the present application.

[0023] The test methods used in the embodiments are all conventional methods unless otherwise specified; and the materials, reagents, etc. used are all commercially available unless otherwise specified.

[0024] In different embodiments of the anti-sugar composition of the present application, the anti-sugar composition is added in an amount of 0.001% to 5%, preferably 0.01% to 5%, and more preferably 0.01% to 3%, according to the form of the oral care product.

[0025] The raw materials of the anti-sugar children toothpaste of the present application include a humectant, a natural antibacterial agent, a thickening agent, a soft abrasive, xylitol, a calcium-containing compound, the anti-sugar composition, a mild surfactant, a fragrance, and deionized water. The content of each raw material ranges from 25% to 28% for the humectant, from 0.1% to 1% for the natural antibacterial agent, from 0.2% to 1% for the thickening agent, from 10% to 25% for the soft abrasive, from 0.01% to 20% for the xylitol, from 0.001% to 5% for the calcium-containing compound, from 0.001% to 5% for the anti-sugar composition, from 0.01% to 2% for the mild surfactant, from 0.5% to 1% for the fragrance, and the balance is water (preferably deionized water).

[0026] The humectant is at least one of sorbitol, glycerol, polyethylene glycol, propylene glycol.

[0027] The natural antibacterial agent is at least one of benzyl alcohol, octanediol, hexanediol, p-hydroxyacetophenone. The natural antibacterial agent belongs to food-grade ingredients, is safe and non-toxic, has good antibacterial and bacteriostatic performance, and can prolong the shelf life of food.

[0028] The thickening agent is at least one of xanthan gum, cellulose gum, carbomer.

[0029] The soft abrasive is silicon dioxide.

[0030] The calcium-containing compound is at least one of soluble calcium or slightly soluble calcium. The soluble calcium or slightly soluble calcium is at least one of calcium lactate, glycerophosphate calcium, and calcium gluconate. The children's anti-sugar toothpaste also adds xylitol, which is a natural plant sweetener and cannot be metabolized by bacteria in the oral cavity, so it will not produce acid to erode the teeth. The combination of xylitol and the calcium-containing compound can inhibit bacterial metabolism while promoting tooth remineralization, thereby resisting bacterial erosion and strengthening the teeth.

[0031] The mild surfactant is at least one of sodium lauroyl sarcosinate, sodium lauroyl glutamate, cocamidopropyl betaine, sodium N-methyl cocoyl taurate, poloxamer, and alkyl glycoside. The mild surfactant cleans the teeth without damaging the oral mucosa, is safe and non-irritating, and can be used alone or in combination with sodium lauryl sulfate.

[0032] The anti-sugar composition comprises 0.001-0.1 parts by weight of glucose oxidase, 0.001-0.1 parts by weight of dextranase, 0.001-10 parts by weight of hydrolyzed wheat protein, 0.01-0.5 parts by weight of arginine, 0.01-0.05 parts by weight of fluoride, and 0.001-0.1 parts by weight of probiotics. The multiple active ingredients in the anti-sugar composition can synergistically inhibit the growth of cariogenic bacteria and other oral pathogenic bacteria, inhibit plaque formation, resist acid erosion of tooth enamel, and promote tooth remineralization. The ratio of the addition amounts of dextranase and glucose oxidase in the anti-sugar composition is 0.001:1-1:1, and the preferred ratio is 0.01:1-1:1. The ratio of the addition amounts of dextranase or glucose oxidase and hydrolyzed wheat protein or fluoride is 0.001:1-1:1, and the preferred ratio is 0.01:1-1:1.

[0033] In the following examples and comparative examples, the performance detection method is as follows:

[0034] (1) Investigation of high-temperature stability of toothpaste formula physical and chemical indicators

[0035] Toothpaste pH reference GB 8372 2017 Toothpaste pH test method.

[0036] Toothpaste stability reference GB 8372 2017 Toothpaste stability test method, with slight modification: take two samples of toothpaste, one is stored at room temperature, the other is placed in a 45±1℃ constant temperature incubator, and is taken out at different time intervals for comparison with the room temperature, to check the stability: compare the appearance and color of the paste with the room temperature stored sample, whether the smell is normal, whether there are abnormal phenomena such as coarsening, water separation, etc., then cut the tube to check whether the paste has abnormal phenomena such as granulation, coarsening, water separation, shell separation, etc., if not, it is considered to be stable.

[0037] Toothpaste consistency method reference GB 8372 2001 Toothpaste consistency test method.

[0038] (2) Toothpaste plaque removal effect evaluation method

[0039] 1) Inoculate Streptococcus mutans into 8mL BHI broth and incubate at 37℃ in aerobic conditions for 20h-24h overnight;

[0040] 2) Take a 24-well plate, add 200 microliters of bacterial solution to each well, and then add 1800 microliters of BHI containing 5% sucrose, and incubate at 37℃ in aerobic conditions for 20h;

[0041] 3) Discard the culture solution with a 5mL pipette, and wash once with 2mL of 0.1mol / L(pH=6.0) PBS along the wall of the well;

[0042] 4) Add 2ml of 37℃ pre-warmed toothpaste solution (10-fold dilution) in a water bath or incubator at 37℃ for 5min;

[0043] 5) Wash with 2mL of PBS for 3 times to remove residual toothpaste solution;

[0044] 6) Dry in a 60℃ oven for 30min;

[0045] 7) Add 2mL of 0.1% crystal violet staining solution to each well and stain for 10min;

[0046] 8) Discard the staining solution and wash with 2mL of PBS for 3 times, and dry at 60℃ for 10-15min;

[0047] 9) Add 2mL of acetone:ethanol=2:8 eluent and measure the absorbance value at 590nm;

[0048] Plaque removal rate = [(blank absorbance - enzyme preparation treatment solution absorbance) / blank absorbance] x 100%.

[0049] (2) Inhibition of in vitro dental plaque glycolysis evaluation method

[0050] Principle: By cultivating dental plaque, observe the pH difference between sample group and control group after sample treatment to see whether the sample group can inhibit the growth of dental plaque.

[0051] Reagent preparation: ① Plaque growth medium: dissolve 6g BBL tryptone peptone, 20g sucrose in 160.8g deionized water, sterilize at 121℃ for 20min and cool down. Add 13.2g fresh saliva before use. ② Glycolysis medium: dissolve 6g BBL tryptone peptone, 10g sucrose in 184g deionized water, sterilize at 121℃ for 20min and cool down. ③ Experimental grouping: sample treatment solution: toothpaste sample is prepared into toothpaste slurry with toothpaste: water = 1:3; negative control: normal saline; positive control: glycolysis medium.

[0052] Operation steps: ① Plaque formation: collect saliva from 3-5 people, mix (about 100mL), add sucrose to the concentration of 0.1% in the saliva, add 5mL of the saliva in a test tube. Insert a cleaned and polished glass rod and soak at 37℃ overnight. ② Plaque growth: collect 13.2g fresh saliva and mix in the growth medium. Take 5mL of the medium in a new test tube, transfer the glass rod treated in ① to the test tube, soak at 37℃ for more than 6h. Then transfer the glass rod to a new test tube containing 5mL of fresh saliva, soak at 37℃ overnight. ③ Add 6-8mL of treatment solution to the test tube. Soak the glass rod treated in ② in the treatment solution for 1s each time, for 60s. Then wash the glass rod in a test tube containing deionized water, 10s each time, for 2 times. ④ Add 5mL of glycolysis medium in a new test tube, soak the glass rod washed in ③ in the test tube, treat at 37℃ for 6h. ⑤ Measure the pH and record.

[0053] Efficacy evaluation: efficacy% compared with the positive control = [1-(average pH value of the positive control- average pH value of the experimental group) / (average pH value of the positive control- average pH value of the negative control group)]x100%

[0054] The above value greater than or equal to 50% indicates effectiveness, i.e. can inhibit in vitro dental plaque glycolysis.

[0055] (3) Method for detecting the stability of biological enzyme activity in toothpaste

[0056] Principle: Dextranase can hydrolyze the α-1,6 glycosidic bond in dextran. The reaction product is isomaltulose. The activity of dextranase is determined by measuring the amount of reducing sugar formed. Llanes method is used to determine reducing sugar.

[0057] Reagent preparation: Substrate solution: Dextran from Leuconostoc dextranicum (Sigma, NO. 31390), dissolve 0.5 g dry (105℃, 4 h) in water and dilute to 50 ml. (Freshly prepared); Reagent A: 8.25 g potassium ferricyanide + 10.6 g anhydrous sodium carbonate, make up to 1 L with water, and store in the dark for 2-3 days. (Valid for 3 months); Reagent B: 25 g potassium iodide + 50 g zinc sulfate heptahydrate + 250 g sodium chloride, make up to 1 L with water (Valid for 3 months); Reagent C: 5 ml acetic acid, dilute to 100 ml with water;

[0058] Determination procedure: Mix 10 ml substrate solution and 4 ml 0.1 M acetic acid buffer in test tube ①, and incubate the mixture at 37℃ for 10 min, then add 1 ml enzyme solution to test tube ① and mix, continue to incubate at 37℃ for 30 min; Prepare another test tube ② in advance, add 5 ml reagent A + 3 ml water for standby. After incubation in test tube ① is completed, take 2 ml reaction solution to test tube ②, and place test tube ② in a boiling water bath for 5 min, then take out test tube ② after cooling, successively add 5 ml reagent B + 3 ml reagent C, mix well, and titrate with 0.01 mol / L sodium thiosulfate, the solution turns milky white and does not fade for 30 s as the end point, record the titration volume.

[0059] Calculation: One unit of dextranase activity is defined as the amount of enzyme that can produce the same reducing power as one micromole of sodium thiosulfate per minute under the above conditions.

[0060] Units / ml = (B-T) x 0.01 x 1000 x (1 / 30) x (15 / 2) x F x N

[0061] B: Blank titration volume (unit: ml)

[0062] T: Titration volume of enzyme sample (unit: ml)

[0063] F: Coefficient of 0.01 mol / L sodium thiosulfate

[0064] N: Dilution factor of enzyme

[0065] (4) Detection method of toothpaste inhibiting oral pathogenic bacteria

[0066] The performance of children's anti-sugar toothpaste in inhibiting oral pathogenic bacteria was evaluated according to the detection method in GB 15979-2002 Appendix C4.

[0067] (5) Detection method of toothpaste inhibiting glycolysis effect

[0068] ① Reagent preparation:

[0069] Plaque growth medium: 6 g BBL Trypticase Soy Broth, 20 g sucrose, dissolved in 160.8 g deionized water, sterilized at 121 °C for 20 min and cooled. 13.2 g fresh saliva was added before use. Glycolysis medium: 6 g BBL Trypticase Soy Broth, 10 g sucrose, dissolved in 184 g deionized water, sterilized at 121 °C for 20 min and cooled. Treatment solution preparation: Toothpaste sample was prepared as a toothpaste slurry with water at a ratio of 1:3, and other antibacterial substances were prepared as solutions at appropriate concentrations.

[0070] ②Glass rod polishing

[0071] New glass rods were polished on a lathe using silicon carbide sandpaper of 240, 320, 400, and 600 grits, starting from 25 mm from one end. After initial polishing, 600 grit sandpaper was used again before each experiment. Negative and positive control groups were set up, with a maximum of 1-2 experimental groups per experiment, and 4 glass rods were needed for each experimental group.

[0072] ③Experimental procedure

[0073] Day 1: Glass rods were cleaned with dilute hydrochloric acid under ultrasonic agitation, rinsed, dried, and polished with 600 grit sandpaper. The glass rods were rinsed with deionized water and dried. Saliva was collected from 3-5 people, mixed (about 100 mL), and sucrose was added to a concentration of 0.1%. 5 mL of the saliva was added to 16 test tubes. The glass rods were inserted and soaked at 37 °C overnight. Plaque growth medium was prepared and sterilized at high temperature for use the next day.

[0074] Day 2: Fresh saliva was collected and added to the growth medium, mixed well. 5 mL of the medium was taken to new test tubes, and the glass rods from the previous day were transferred to new 16 test tubes and soaked at 37 °C for more than 6 hours. Then they were transferred to 16 test tubes containing 5 mL of fresh saliva each and soaked at 37 °C overnight. Glycolysis medium was prepared and sterilized at high temperature for use the next day.

[0075] Day 3: The treatment solution was prepared and added to 4 test tubes, ensuring that it fully covered the plaque on the glass rods (about 6-8 mL). 10 mL of deionized water was added to 32 test tubes. The glass rods were removed from the 37 °C constant temperature incubator and soaked in the treatment solution for 60 seconds each time. Then the glass rods were washed in the deionized water test tubes, 10 seconds each time, for a total of 2 washes. 5 mL of glycolysis medium was added to 16 test tubes, and the washed glass rods were soaked in the glycolysis test tubes at 37 °C for 6 hours. The pH of the experimental and control groups was measured and recorded. The glass rods were placed in a beaker and soaked in 1 M HCl for cleaning and recovery.

[0076] ④Efficacy evaluation

[0077] Efficacy % compared with positive control = [1- (pH average of positive control - pH average of experimental group) / (pH average of positive control - pH average of negative control group)] x 100%

[0078] The above value greater than or equal to 50% indicates effectiveness.

[0079] (6) In vitro anti-acid erosion model evaluation method

[0080] Principle: Use bovine enamel to simulate human teeth for toothpaste slurry immersion treatment, and use glycolysis liquid to erode the teeth, and observe whether there is a significant difference in the hardness change of the bovine tooth sample before and after the treatment compared with the control to evaluate the anti-acid erosion ability.

[0081] Reagent preparation: ① Artificial saliva: 200 g / L MUCIN TYPE II, 0.38 g / L NaCl, 0.183 g / L CaCl2, 0.528 g / L KH2PO4, 1.114 g / L KCl. First, stir the MUCIN with hot water at 80°C until it is completely dissolved, then add the other ingredients in turn, stir and dissolve, and finally adjust the pH to 7.0 with 1 mol / L KOH solution; ② Glycolysis liquid: 6 g of pancreatic tryptone soy broth, 20 g of sucrose, dissolved in 160.8 g of deionized water, sterilized at 121°C for 20 min and cooled. Collect 3-4 fresh saliva of human, add 13.2 g of fresh saliva before use. Put the culture medium with saliva into a 37°C incubator for 18-24 h, and measure the pH value at about 4.00. Finally, sterilize at 121°C for 30 min, and cool for standby; ③ Sample treatment liquid: toothpaste sample is prepared into a toothpaste slurry with toothpaste: water = 1:3.

[0082] Operation steps:

[0083] 1) Select the enamel sample with a hardness value of (230-360) kg / mm 2 ;

[0084] 2) Perform in vitro circulation treatment on the selected enamel sample, as shown in Table 1:

[0085] Table 1 Circulation treatment

[0086]

[0087] Note: After 6 times of treatment of serial numbers 1-3, perform the treatment of serial number 4.

[0088] 3) Test the hardness value of the in vitro circulation treated enamel sample, recorded as HV 酸蚀前 , soak in glycolysis liquid for 30 min, and measure the hardness value, recorded as HV 酸蚀后 .

[0089] 4) Remineralization treatment: the acid-etched enamel samples were treated again in vitro circulation, operation same as 2), then test the hardness value, recorded as HV 再矿化 .

[0090] Efficacy evaluation:

[0091] 1) Anti-erosion ability evaluation: calculate the change of microhardness value before and after acid-etching ΔHV1=HV 酸蚀前 -HV 酸蚀后 , independent sample t test was used. If the ΔHV value of the sample group was less than that of the control group, and the comparison of the ΔHV values of the two groups was statistically significant (P<0.05), it was considered that the sample group could resist the erosion of the enamel caused by the acid produced by glycolysis in the oral cavity.

[0092] 2) Evaluation of the ability to promote remineralization: calculate the change of hardness value before and after remineralization ΔHV2=HV 再矿化 -HV 酸蚀后 , independent sample t test was used. If the ΔHV value of the sample group was greater than that of the control group, and the comparison of the ΔHV values of the two groups was statistically significant (P<0.05), it was considered that the sample group could promote the remineralization ability of the enamel eroded by glycolysis in the oral cavity.

[0093] The composition of the anti-sugar children toothpaste of Examples 1-10 and Comparative Examples 1-11 is shown in Table 2.

[0094] The preparation method of the anti-sugar children toothpaste of Examples 1-10 and Comparative Examples 1-11 is as follows:

[0095] Sodium pyrophosphate, xylitol, calcium lactate, sodium fluoride were added to deionized water, stirred until completely dissolved, then sorbitol, glycerol, polyethylene glycol-8, benzyl alcohol were added, stirred for 5 min to prepare the liquid phase. Silica, xanthan gum were added to a container and stirred uniformly, then the powder was added to the liquid phase, stirred for 20 min. Finally, essence, cocamidopropyl betaine, saliva lactobacillus, glucose oxidase, arginine, hydrolyzed wheat protein were added, stirred for 15 min and degassed to prepare the children anti-sugar toothpaste.

[0096] Table 2 Anti-sugar children toothpaste components

[0097]

[0098] Table 2 Anti-sugar children toothpaste components (continued)

[0099]

[0100] The results of the stability test of the physicochemical indicators of the anti-sugar children toothpaste of Examples 1-5 are shown in Table 3:

[0101] Table 3 Stability test of physicochemical indicators of anti-sugar children toothpaste

[0102]

[0103] As shown in Table 3, the anti-sugar children toothpaste of Examples 1-5 was detected for various indicators after accelerated aging at 45℃ for 3 months. The detection results showed normal, the anti-sugar children toothpaste of Examples 1-5 had good overall stability, normal physical and chemical indicators, good compatibility of formula components, no water oil separation phenomenon, and excellent quality.

[0104] The plaque removal effect of the anti-sugar children toothpaste of Examples 1-5 is shown in Table 2. Figure 1 The anti-sugar children toothpaste of Examples 1-5 all had obvious plaque removal effect, and the plaque removal effect was derived from the decomposition of dental plaque biofilm by the biological enzyme in combination with other components, and the plaque removal effect was obviously improved with the increase of the concentration of the biological enzyme.

[0105] The detection results of the biological enzyme activity stability of the anti-sugar children toothpaste of Examples 1-5 are shown in Table 4.

[0106] Table 4 Enzyme activity of anti-sugar children toothpaste

[0107]

[0108]

[0109] After two months of high temperature, the average enzyme activity of the anti-sugar children toothpaste of Examples 1-5 was about 75%, and the residual activity was increased with the increase of the enzyme concentration, and the highest was 83%, indicating that the biological enzyme could maintain high enzyme activity in the formula system of the anti-sugar toothpaste and play an active role.

[0110] The effect of the anti-sugar children toothpaste of Examples 1-5 and Comparative Examples 1-11 on inhibiting oral pathogenic bacteria is shown in Table 5.

[0111] Table 5 Bacteriostatic rate of anti-sugar children toothpaste for inhibiting oral pathogenic bacteria

[0112] Bacteriostatic rate Streptococcus mutans % Porphyromonas catoniae % Fusobacterium nucleatum % Example 1 ＞99.8 ＞99.8 ＞97.5 Example 2 ＞99.8 ＞99.8 ＞97.5 Example 3 ＞99.8 ＞99.8 ＞97.5 Example 4 ＞99.8 ＞99.8 ＞97.5 Example 5 ＞99.8 ＞99.8 ＞97.5 Comparative Example 1 66 67 58 Comparative Example 2 60 59 65 Comparative Example 3 76 82 75 Comparative Example 4 31 35 28 Comparative Example 5 26 29 31 Comparative Example 6 77 86 89 Comparative Example 7 83 90 88 Comparative Example 8 92 87 66 Comparative Example 9 72 76 78 Comparative Example 10 91 92 86 Comparative Example 11 89 90 92

[0113] The anti-sugar children toothpaste of Examples 1-5 can effectively inhibit the growth of oral pathogenic bacteria.

[0114] The effect of the anti-sugar children toothpaste of Example 5 on inhibiting glycolysis is shown in Table 6. Figure 2 The plaque treated with the anti-sugar children toothpaste of Example 5 was inhibited and could not grow and reproduce. The plaque treated with the toothpaste without anti-sugar composition still grew vigorously and showed turbid solution and acidic solution.

[0115] The results of the evaluation of the ability of the sugar-resistant children's toothpaste of Examples 1 to 5 and Comparative Examples 1 to 11 to inhibit the acid production of dental plaque are shown in Table 6.

[0116] Table 6 Evaluation of the ability of the sugar-resistant children's toothpaste to inhibit the acid production of dental plaque

[0117]

[0118]

[0119] As shown in Table 6, in the test of the evaluation of the ability to inhibit the acid production of dental plaque, the sugar-resistant composition in the sugar-resistant children's toothpaste of Examples 1 to 5 had a synergistic effect, and could synergistically inhibit the acid production of dental plaque, and when the concentration was increased, the effect of inhibiting the acid production of dental plaque was also obviously increased. At the same time, when the single component in the sugar-resistant children's toothpaste of Comparative Examples 1 to 5 was compared, the effect of the single component to inhibit the acid production of dental plaque was poor, far less than the synergistic inhibition in the multi-component. Biological enzymes such as dextranase, glucose oxidase, etc. have the effect of decomposing dental plaque biofilm and reducing bacterial adhesion, and can inhibit bacterial multiplication to a certain extent, and experimental data show that hydrolyzed wheat protein as a kind of wheat polypeptide also has the effect of inhibiting bacterial multiplication. Sodium fluoride and arginine have weak bacteriostatic ability on bacteria, and thus have weak ability to inhibit the acid production of dental plaque. It can be seen that the sugar-resistant composition in the sugar-resistant children's toothpaste of Examples 1 to 5 has the sugar-resistant effect of inhibiting the acid production of dental plaque. In addition, through the comparison of each example and Comparative Examples 6 to 11, it can be seen that the synergistic effect of various compositions can obviously promote the inhibition of the acid production of dental plaque, and has a synergistic effect.

[0120] The results of the evaluation of the acid-erosion resistance and the ability to promote remineralization of the sugar-resistant children's toothpaste of Examples 1 to 5 and Comparative Examples 1 to 11 are shown in Tables 7 and 8.

[0121] Table 7 Evaluation of the acid-erosion resistance of the children's sugar-resistant toothpaste

[0122]

[0123]

[0124] Table 8 Evaluation of the ability of the children's sugar-resistant toothpaste to promote remineralization

[0125]

[0126] As can be seen from Table 7 and Table 8, the anti-erosion ability and the ability to promote enamel remineralization of the sugar-resistant children's toothpaste of Examples 1-5 have significant differences compared with the negative control group. Through comparison of the single components of Comparative Examples 1-5, the single components of probiotics, dextranase and dextranase cannot resist tooth erosion and promote enamel remineralization, but fluoride, hydrolyzed wheat protein and arginine can resist tooth erosion and promote enamel remineralization, but the anti-erosion and tooth remineralization ability of hydrolyzed wheat protein is obviously better than that of sodium fluoride and arginine; in addition, the effect of the sugar-resistant children's toothpaste of Examples 1-5 is obviously lower than the anti-erosion and remineralization ability of the sugar-resistant composition of multiple components. At the same time, by comparing with Comparative Examples 6-11, it can be seen that the sugar-resistant children's toothpaste lacking any single sugar-resistant component has lower anti-erosion and remineralization ability than the sugar-resistant children's toothpaste containing all the sugar-resistant components (Examples 1-5). As can be seen from Table 7, the anti-erosion ability of the sugar-resistant children's toothpaste of Examples 1-5 is increasing with the increase of the components and concentration of the sugar-resistant active ingredients. At the same time, as can be seen from Table 8, the ability to promote enamel remineralization of the sugar-resistant children's toothpaste of Examples 1-5 is also increasing with the increase of the components and concentration of the sugar-resistant active ingredients, but the enamel hardness is relatively stable after increasing to a certain level, and the remineralization effect is not obvious (Examples 3-5). Therefore, the glucose oxidase, dextranase, fluoride, hydrolyzed wheat protein, arginine and probiotics in the sugar-resistant composition play a synergistic effect, and jointly play the sugar-resistant effect.

[0127] In summary, the present application provides a sugar-resistant composition comprising glucose oxidase, dextranase, hydrolyzed wheat protein, sodium fluoride, probiotics and arginine, and a sugar-resistant children's toothpaste comprising the sugar-resistant composition, which has a sugar-resistant effect, specifically a multi-path sugar-resistant effect of inhibiting plaque acid production, resisting enamel erosion and promoting enamel remineralization. The sugar-resistant composition can protect children's teeth from sugar through multiple paths, inhibit plaque formation, remove dental plaque, resist tooth demineralization caused by bacterial glycolysis acid production due to sugar in food, promote tooth remineralization, and maintain oral health.

[0128] The glucose oxidase in the anti-sugar composition of the present application is an aerobic dehydrogenase prepared by fermentation of Aspergillus niger and the like. It is non-toxic and has no side effects on the human body, and has the functions of removing glucose, deoxidizing, sterilizing and the like. The glucose oxidase has the advantages of catalytic specificity, high activity and high catalytic efficiency, and can decompose the glucan in the food residues adhering to the teeth, thereby inhibiting the formation of dental plaque. In addition, the glucose oxidase has good adhesion on the teeth and can continuously take effect. The hydrogen peroxide generated in the process of the glucose oxidase decomposing glucose is gradually released, and will not suddenly produce too much to stimulate other tissues in the oral cavity, and can safely and effectively remove stains and dental plaque without damaging the enamel and irritating the gums, and can efficiently whiten the teeth in a short time. The dextranase in the anti-sugar composition of the present application is a kind of glucanase, which can cut the alpha-1, 6-glycosidic bond and degrade glucan. The biofilm of dental plaque is mainly composed of extracellular polysaccharide, and the main component is glucan. Studies have shown that dextranase can inhibit the initial formation of oral pathogenic bacterial biofilm, reduce bacterial adhesion, change the structure of biofilm, and destroy the crosslinking between bacterial colonies. Therefore, dextranase can effectively decompose dental plaque biofilm, thereby inhibiting the formation of dental plaque. The present application overcomes the problem that the stability and activity of biological enzymes in toothpaste are easily changed.

[0129] The anti-sugar composition of the present application includes oral probiotics. Probiotics are a general term for microorganisms that are beneficial to the host, including active strains and inactivated strains, and can help the human body adjust or maintain the microecological balance, prevent diseases, and promote the health of the body. The probiotics are at least one of Bifidobacterium adolescentis, Bifidobacterium lactis, Bifidobacterium bifidum, Lactobacillus crispatus, Bifidobacterium longum, Bifidobacterium breve, Lactobacillus bulgaricus, Lactobacillus gasseri, Lactobacillus helveticus, Lactobacillus johnsonii, Lactobacillus plantarum, Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus salivarius and Streptococcus thermophilus. In view of the defect that probiotics cannot be stored at room temperature, the present application uses inactivated freeze-dried powder of probiotics to replace live bacteria. After the probiotics enter the oral cavity with toothpaste, they occupy the binding points on the oral mucosa by adhesion, thereby reducing the colonization of pathogenic bacteria in the oral environment, inhibiting the growth and reproduction of pathogenic bacteria, and reducing the number of harmful bacteria such as cariogenic bacteria, periodontal pathogenic bacteria and oral Candida. Probiotics can also self-aggregate and co-aggregate with harmful bacteria to form a biofilm, which is wrapped and taken out of the oral cavity, thereby inhibiting the growth of pathogenic bacteria and forming a barrier to prevent pathogenic bacteria from colonizing and infecting.

[0130] The hydrolyzed wheat protein in the anti-sugar composition of the present application uses wheat protein as raw material, and adopts chemical hydrolysis, microbial fermentation, protease hydrolysis and other methods to obtain small molecular polypeptide substances, which have the characteristics of good water solubility, dispersion stability, easy absorption and strong biological activity. The main component of the hydrolyzed wheat protein is wheat peptide, which can chelate mineral elements, promote the absorption and utilization of animals to mineral elements. The hydrolyzed wheat protein can chelate calcium ions and adhere to the surface of hydroxyapatite (the main component of tooth enamel), increase the hardness of teeth, promote tooth remineralization and repair acid-eroded teeth. In addition, the hydrolyzed wheat protein has a synergistic effect when used with fluoride, which can further enhance the remineralization effect of fluoride and reduce the toxic side effects of fluoride.

[0131] The arginine in the anti-sugar composition of the present application plays an important role in the human body. As a component of salivary mucin, it can adhere to the surface of teeth with the semi-permeable membrane function of salivary mucin, and further regulate the balance of oral flora and the dynamic balance between ions. At the same time, arginine can buffer and regulate the pH value of saliva by participating in the hydrogen-producing Neisseria metabolism, and due to its good solubility, it is easy to adsorb on the tooth surface and dentin tubules, thereby promoting the deposition of calcium and phosphorus ions on the surface of dentin and in the dentin tubules, and promoting tooth remineralization. In addition, arginine has the ability to promote hydroxyapatite deposition and close dentin tubules, thereby achieving the effect of anti-sensitivity.

[0132] The product of the present application adopts a scientific use method, and repeated use within a period of several days can effectively resist sugar, including helping to inhibit oral bacteria, remove dental plaque biofilm, inhibit plaque formation, resist bacterial glycolysis to produce acid, and at the same time resist tooth demineralization caused by glycolysis to produce acid, promote tooth remineralization, etc. Specifically, by decomposing glucose through anti-sugar ingredients, reducing the source of oral bacterial metabolism, removing dental plaque polysaccharides to reduce soft plaque, reducing dental plaque, inhibiting the growth of acid-producing bacteria, and at the same time being able to remove dental plaque biofilm and neutralize the acid produced by bacteria, further enhancing the hardness of teeth, promoting tooth remineralization, inhibiting tooth demineralization, and achieving the effect of inhibiting glycolysis to acid-erode teeth.

[0133] Finally, it should be noted that the above examples are only used to illustrate the technical solutions of the present application and not to limit the protection scope of the present application. Although the present application has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present application can be modified or replaced by equivalents without departing from the essence and scope of the technical solutions of the present application.

Claims

1. An anti-glycating composition comprising hydrolyzed wheat protein, characterized in that, The anti-sugar composition comprises, by mass parts, the following components: hydrolyzed wheat protein 0.001-10 parts, glucose oxidase 0.001-0.1 part, dextranase 0.001-0.1 part, arginine 0.01-0.5 part, fluoride 0.01-0.05 part, and probiotic 0.001-0.1 part.

2. The anti-sugar composition of claim 1, wherein, The ratio of the two added amounts of the dextranase and the glucose oxidase is 0.001:1-1:

1.

3. The anti-sugar composition of claim 1, wherein, The ratio of the two added amounts of the dextranase or the glucose oxidase and the hydrolyzed wheat protein or the fluoride is 0.001:1-1:

1.

4. Use of the anti-sugar composition according to any one of claims 1-3 in the preparation of an oral care product.

5. An anti-sugar oral care product characterized in that, The anti-sugar oral care product comprises, by mass percentage, 0.001%-5% of the anti-sugar composition according to any one of claims 1-3.

6. The anti-glycated oral care product of claim 5, wherein, The anti-sugar oral care product is an anti-sugar children's toothpaste.

7. The anti-glycated oral care product of claim 6, wherein, The anti-sugar children's toothpaste further comprises, by mass percentage, the following components: humectant 25%-28%, natural antibacterial agent 0.1%-1%, thickening agent 0.2%-1%, soft abrasive 10%-25%, xylitol 0.01%-20%, calcium-containing compound 0.001%-5%, mild surfactant 0.01%-2%, essence 0.5%-1%, and the balance being water.

8. The anti-glycated oral care product of claim 7, wherein, The natural antibacterial agent is at least one of benzyl alcohol, caprylyl glycol, hexylene glycol, and p-hydroxyacetophenone.

9. The anti-glycated oral care product of claim 7, wherein, The calcium-containing compound is at least one of soluble calcium or slightly soluble calcium.

10. The anti-glycated oral care product of claim 7, wherein, The mild surfactant is at least one of sodium lauroyl methyl amino acid, sodium lauroyl glutamate, cocamidopropyl betaine, sodium N-methyl cocoyl taurate, poloxamer, and alkyl polyglycoside.

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