A stable oral care composition containing high purity bee oligopeptides

Abstract

The application discloses a kind of stable oral care compositions containing high-purity bee oligopeptide, belongs to oral care technical field, specifically related to a kind of oral care compositions including bee oligopeptide complex and regulator, the purity of bee oligopeptide in bee oligopeptide complex is more than 95%, the mass ratio of bee oligopeptide complex and regulator is 0.02-1:10-8140.The application is prepared to obtain oral care compositions by using bee oligopeptide complex, regulator and water mixing, bee oligopeptide complex is used after being compounded by certain proportion by natural bee oligopeptide and synthetic bee oligopeptide, the use of regulator can adjust the activity of water in oral care compositions, improve the stability of bee oligopeptide.In the application, additive can also be added, the use of additive and regulator together can also adjust the activity of water in oral care compositions, and oral care compositions with different components can be prepared, which can expand the use of oral care compositions.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of oral care, and particularly relates to an oral care composition containing high-purity melittin. BACKGROUND

[0002] Oral microorganisms are the most critical cause of oral diseases. Oral microbial communities are composed of more than 700 gram-negative and gram-positive bacteria, including aerobic and anaerobic bacteria. Microorganisms on the surface of the oral cavity and teeth often exist in the form of biofilms. These bacteria / biofilms can easily cause various oral diseases, such as halitosis, dental caries, gingivitis, and even tooth loss.

[0003] Antibacterial peptides can effectively and broadly inhibit the growth of bacteria and their biofilms, and will not cause microbial resistance or induce superbugs. Among them, melittin has attracted widespread attention due to its natural source, low effective concentration, and anti-inflammatory activity. Melittin is also called bee venom peptide. The most abundant bioactive polypeptide in bee venom is melittin, which accounts for about 40-60% of the dry weight of bee venom. Melittin is a polypeptide composed of 26 amino acid residues, with a molecular weight of 2840. The pure product is a white powder, and the aqueous solution is strongly alkaline with a pH of about 10.0. Currently, people mainly obtain melittin by separation and extraction from bee venom or chemical synthesis.

[0004] Melittin is a naturally occurring antibacterial peptide that has resistance function to harmful life forms or pathogens such as bacteria, fungi, parasites, and viruses. Melittin has sufficient positive charge and is often accompanied by hydrophobicity, which can bind to biofilms containing negative electricity under the action of static electricity, penetrate and destroy the membrane structure to cause cell death. Unlike the single-target bactericidal principle of traditional antibiotics, antibacterial peptides can destroy pathogens at multiple targets, greatly reduce the generation of drug-resistant bacteria, and have broad-spectrum antibacterial properties, making them one of the best choices to replace antibiotics in the future.

[0005] Naturally isolated melittin contains a relatively high phospholipase A2 (PLA2), which has significant cytotoxicity and significant hemolytic activity. Melittin often exists in the form of a tetramer. The tetramer structure is similar to the structure of phospholipase A2, and the alkalinity and hydrophobicity are also very close, making it difficult to purify and separate. The synergistic effect of melittin and phospholipase A2 can easily cause hemolysis and trigger immune responses in the body. Naturally occurring melittin is easily inactivated in different solvents such as water, ethanol-water mixtures, etc., and is also easily affected by concentration, light, temperature, and pH during long-term storage, resulting in a decrease in activity or inactivation. SUMMARY

[0006] The present application aims to provide an oral care composition containing high-purity melittin with good stability, good inhibitory effect on Streptococcus mutans, and good inhibitory effect on Porphyromonas gingivalis.

[0007] The technical scheme adopted by the present application to achieve the above-mentioned purpose is:

[0008] An oral care composition comprises: a bee oligopeptide complex and a modulator mixed together, the purity of the bee oligopeptide in the bee oligopeptide complex is more than 95%, and the mass ratio of the bee oligopeptide complex to the modulator is 0.02-1:10-8140. It is very difficult to purify the naturally occurring bee oligopeptide. The dry weight of natural bee venom contains 40-60% bee oligopeptide. The dry weight usually exists in the form of a tetramer, and the molecular weight and the allergic component phosphatase are very close. The basicity is also similar. Therefore, it is relatively difficult to purify and separate the bee oligopeptide. The cytotoxicity of the unpurified melittin is about 1 ppm. The purity of the synthetic bee oligopeptide is higher. By compounding the natural bee oligopeptide and the synthetic bee oligopeptide, the stability of the bee oligopeptide can be improved while reducing the toxicity of the natural bee oligopeptide. In the oral composition, the modulator includes short-chain or long-chain alcohol substances, and the combination of the alcohol substances can reduce the water activity. In the present application, the bee oligopeptide complex and the modulator are compounded, the water activity is adjusted to less than 0.90 when water is used as the solvent, and the purity of the bee oligopeptide in the bee oligopeptide complex is more than 95%. Under the specific water activity and the purity of the bee oligopeptide in the bee oligopeptide complex, the stability of the bee oligopeptide in the oral care composition can be improved.

[0009] Preferably, the modulator includes at least one of glycerol, PEG, ethanol, sorbitol, propylene glycol and D-panthenol.

[0010] More preferably, the mass ratio of the bee oligopeptide complex to glycerol is 0.02-1:300-2000; or,

[0011] The mass ratio of the bee oligopeptide complex to PEG is 0.02-1:200-1500; or,

[0012] The mass ratio of the bee oligopeptide complex to ethanol is 0.02-1:1000-2000; or,

[0013] The mass ratio of the bee oligopeptide complex to sorbitol is 0.02-1:1000-2000; or,

[0014] The mass ratio of the bee oligopeptide complex to propylene glycol is 0.02-1:100-600; or,

[0015] The mass ratio of the bee oligopeptide complex to D-panthenol is 0.02-1:10-40.

[0016] Preferably, the oral care composition further comprises an additive.

[0017] More preferably, the additive comprises a mannosamido compound having a mannosyl group and an amino compound group. The present application finds that the use of the additive, in combination with the regulator, can also affect the water activity in the oral care composition, and the stability of the propolis oligopeptide in the obtained oral care composition can be improved when the mannosamido compound is used as the additive.

[0018] Further more preferably, the content of the mannosamido compound in the oral care composition is 0.02-0.12wt%.

[0019] More preferably, in the preparation of the mannosamido compound, first, the mannosyl chloride is prepared by reacting the mannuronic acid and dichlorosulfoxide, and then the mannosamido compound is obtained by reacting the mannosyl chloride with the amino compound.

[0020] More preferably, in the preparation of the mannosyl chloride, the mannuronic acid is added to a solvent, then dichlorosulfoxide is added at 0-10℃, and the reaction is carried out at 20-40℃ for 2-8h. After the reaction is completed, the solvent and dichlorosulfoxide are removed by rotary evaporation to obtain the mannosyl chloride.

[0021] More preferably, in the preparation of the mannosyl chloride, the solvent is dichloromethane, the use amount of the mannuronic acid is 10-30wt% of the dichloromethane, and the use amount of the dichlorosulfoxide is 60-80wt% of the mannuronic acid.

[0022] More preferably, in the preparation of the mannosamido compound, the amino compound is added to a basic solution, then the mannosyl chloride is added, and the reaction is carried out at 20-40℃ for 4-16h. During the reaction, the pH is controlled at 9-10. After the reaction is completed, the pH is adjusted to 1-2, the precipitate is precipitated and filtered, washed and dried to obtain the mannosamido compound.

[0023] More preferably, in the preparation of the mannosamido compound, the basic solution is a sodium hydroxide solution, and the content of sodium hydroxide in the sodium hydroxide solution is 5-20wt%.

[0024] More preferably, in the preparation of the mannosamido compound, the amino compound is glycyl tyrosine, and the use amount of the amino compound is 5-20wt% of the sodium hydroxide solution.

[0025] More preferably, in the preparation of the mannosamido compound, the use amount of the mannosyl chloride is 100-300wt% of the amino compound.

[0026] More preferably, the additive comprises at least one of anhydrous citric acid, hydrogenated castor oil, essence, xanthan gum, cellulose gum, sodium saccharin, sodium fluoride, methylparaben, hydrous silica, sodium lauryl sulfate, cocamidopropyl betaine, sucralose, glycolipid surfactant, citric acid monohydrate, sodium benzoate, PEG 40, and inorganic salt.

[0027] More preferably, the inorganic salt comprises at least one of sodium chloride, sodium hydroxide, sodium carbonate, sodium bicarbonate, potassium chloride, potassium hydroxide, potassium carbonate, and potassium bicarbonate.

[0028] Preferably, the mass ratio of the bee oligopeptide complex and the xanthan gum is 0.02-1:30-120; or,

[0029] The mass ratio of the bee oligopeptide complex and the cellulose gum is 0.02-1:30-120; or,

[0030] The mass ratio of the bee oligopeptide complex and the sodium saccharin is 0.02-1:20-70; or,

[0031] The mass ratio of the bee oligopeptide complex and the sodium fluoride is 0.02-1:10-50; or,

[0032] The mass ratio of the bee oligopeptide complex and the methylparaben is 0.02-1:5-20; or,

[0033] The mass ratio of the bee oligopeptide complex and the hydrous silica is 0.02-1:1000-3000; or,

[0034] The mass ratio of the bee oligopeptide complex and the sodium lauryl sulfate is 0.02-1:100-400; or,

[0035] The mass ratio of the bee oligopeptide complex and the cocamidopropyl betaine is 0.02-1:30-140; or,

[0036] The mass ratio of the bee oligopeptide complex and the sucralose is 0.02-1:0.2-0.8; or,

[0037] The mass ratio of the bee oligopeptide complex and the glycolipid surfactant is 0.02-1:7-30; or,

[0038] The mass ratio of the bee oligopeptide complex and the citric acid monohydrate is 0.02-1:10-40; or,

[0039] The mass ratio of the bee oligopeptide complex and the sodium benzoate is 0.02-1:25-100; or,

[0040] The mass ratio of the bee oligopeptide complex and the inorganic salt is 0.02-1:2-8; or,

[0041] The mass ratio of the bee oligopeptide complex and the anhydrous citric acid is 0.02-1:60-240; or,

[0042] The mass ratio of the bee oligopeptide complex and the hydrogenated castor oil is 0.02-1:40-160; or,

[0043] The mass ratio of the bee oligopeptide complex and the essence is 0.02-1:10-40.

[0044] Preferably, the oral care composition comprises: the bee oligopeptide complex, the regulator, and water.

[0045] More preferably, the bee oligopeptide complex comprises natural bee oligopeptide and synthetic bee oligopeptide, and the natural bee oligopeptide and the synthetic bee oligopeptide in the bee oligopeptide complex are mixed in a mass ratio of 1:4-10.

[0046] More preferably, the content of the bee oligopeptide complex in the oral care composition is 0.0002-0.01wt%.

[0047] More preferably, the content of the regulator in the oral care composition is 0.1-81.4wt%.

[0048] More preferably, the regulator comprises at least one of glycerol, PEG, ethanol, sorbitol, propylene glycol, and D-panthenol.

[0049] More preferably, the content of glycerol in the oral care composition is 3-20wt%.

[0050] More preferably, the content of PEG in the oral care composition is 2-15wt%.

[0051] More preferably, the content of ethanol in the oral care composition is 10-20wt%.

[0052] More preferably, the content of sorbitol in the oral care composition is 10-20wt%.

[0053] More preferably, the content of propylene glycol in the oral care composition is 1-6wt%.

[0054] More preferably, the content of D-panthenol in the oral care composition is 0.1-0.4wt%.

[0055] More preferably, the oral care composition further comprises an additive, and the additive comprises at least one of anhydrous citric acid, hydrogenated castor oil, essence, xanthan gum, cellulose gum, sodium saccharin, sodium fluoride, hydroxybenzoate, hydrated silica, sodium lauryl sulfate, cocamidopropyl betaine, sucralose, glycolipid, monohydrate citric acid, sodium benzoate, and inorganic salt.

[0056] More preferably, the content of xanthan gum in the oral care composition is 0.3-1.2wt%.

[0057] More preferably still, the cellulose gum is present in the oral care composition in an amount of 0.3 to 1.2 wt%.

[0058] More preferably still, the sodium saccharin is present in the oral care composition in an amount of 0.2 to 0.7 wt%.

[0059] More preferably still, the sodium fluoride is present in the oral care composition in an amount of 0.1 to 0.5 wt%.

[0060] More preferably still, the methylparaben is present in the oral care composition in an amount of 0.05 to 0.2 wt%.

[0061] More preferably still, the hydrous silica is present in the oral care composition in an amount of 10 to 30 wt%.

[0062] More preferably still, the sodium lauryl sulfate is present in the oral care composition in an amount of 1 to 4 wt%.

[0063] More preferably still, the cocamidopropyl betaine is present in the oral care composition in an amount of 0.3 to 1.4 wt%.

[0064] More preferably still, the sucralose is present in the oral care composition in an amount of 0.002 to 0.008 wt%.

[0065] More preferably still, the sugar lipid is present in the oral care composition in an amount of 0.07 to 0.3 wt%. The sugar lipid is RhanceOne sugar lipid.

[0066] More preferably still, the citric acid monohydrate is present in the oral care composition in an amount of 0.1 to 0.4 wt%.

[0067] More preferably still, the sodium benzoate is present in the oral care composition in an amount of 0.25 to 1 wt%.

[0068] More preferably still, the inorganic salt is present in the oral care composition in an amount of 0.02 to 0.08 wt%.

[0069] More preferably still, the anhydrous citric acid is present in the oral care composition in an amount of 0.6 to 2.4 wt%.

[0070] More preferably still, the hydrogenated castor oil is present in the oral care composition in an amount of 0.4 to 1.6 wt%.

[0071] More preferably still, the flavor is present in the oral care composition in an amount of 0.1 to 0.4 wt%.

[0072] More preferably, the oral care composition further comprises ascorbyl glycerin, and the content of ascorbyl glycerin in the oral care composition is 0.01-0.05 wt%. After the regulator is applied to the oral care composition, the ascorbyl glycerin can be further added after the use of the mannosyl amino compound, and the use of the mannosyl amino compound and the ascorbyl glycerin together can improve the stability of the bee oligopeptide in the obtained oral care composition under a specific water activity and the purity of the bee oligopeptide in the bee oligopeptide complex.

[0073] The application discloses the use of the above-mentioned oral care composition in the preparation of an oral care product and / or mouthwash and / or toothpaste.

[0074] The application adopts the bee oligopeptide complex, the regulator and water to prepare the oral care composition, the bee oligopeptide complex is prepared by compounding the natural bee oligopeptide and the synthetic bee oligopeptide in a certain proportion, and the use of the regulator can regulate the water activity in the oral care composition and improve the stability of the bee oligopeptide. In the application, the additive can also be added, and the use of the additive and the regulator together can also regulate the water activity in the oral care composition, so that the oral care composition with different components can be prepared, and the use of the oral care composition can be expanded. Therefore, the application has the following beneficial effects: the bee oligopeptide has good stability, good inhibitory effect on Streptococcus mutans and good inhibitory effect on Porphyromonas gingivalis. Therefore, the application is an oral care composition containing high-purity bee oligopeptide with good stability, good inhibitory effect on Streptococcus mutans and good inhibitory effect on Porphyromonas gingivalis. BRIEF DESCRIPTION OF DRAWINGS

[0075] Figure 1 The infrared spectrum of the mannosyl amino compound.

[0076] Figure 2 The bee oligopeptide loss rate graph of the oral care compositions of Examples 1-7.

[0077] Figure 3 The bee oligopeptide loss rate graph of the oral care compositions of Examples 8-15.

[0078] Figure 4 The bee oligopeptide loss rate graph of the oral care compositions of Examples 16-23. DETAILED DESCRIPTION

[0079] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0080] The concepts involved in this application will first be described with reference to the accompanying drawings. It should be noted that the following descriptions of various concepts are only for the purpose of making the content of this application easier to understand and do not constitute a limitation on the scope of protection of this application; furthermore, the embodiments and features in the embodiments of this application can be combined with each other unless otherwise specified. This application will now be described in detail with reference to the accompanying drawings and embodiments.

[0081] Natural bee oligopeptides, purchased from Sigma Aldrich, catalog number M7391, with a purity of 60% as determined by HPLC.

[0082] The synthetic bee oligopeptide was synthesized by Zhejiang Paipai Biotechnology Co., Ltd., and its purity was 99% as determined by HPLC.

[0083] Bee oligopeptide complex 1 is a mixture of natural bee oligopeptides and synthetic bee oligopeptides in a mass ratio of 1:9, with a purity of 95.1%.

[0084] Bee oligopeptide complex 2 is a mixture of natural bee oligopeptides and synthetic bee oligopeptides in a mass ratio of 1:4, with a purity of 91.2%.

[0085] Antimicrobial test of Streptococcus mutans

[0086] Streptococcus mutans ( Streptococcus mutans The accession number for this is ATCC 700610.

[0087] The culture conditions for Streptococcus mutans were 80% N2 and 20% CO2, and cultured at 37°C.

[0088] Sample preparation: Add 0.01g of sample to 10mL of BHIA medium, vortex until evenly dispersed, and then serially dilute to the corresponding test concentration to obtain the test sample. The sample is a synthetic bee oligopeptide with a purity of 99%.

[0089] Preparation of Streptococcus mutans suspension: Take activated Streptococcus mutans strain, add 5 mL of PBS to initially prepare a bacterial suspension, and adjust the initial Streptococcus mutans concentration to 1×10⁻⁶. 8 CFU / mL, prepare and use immediately.

[0090] Bacteriostatic test of Streptococcus mutans: In a 96-well plate, 180 μL of test sample at different concentrations was added, and then 20 μL of Streptococcus mutans suspension was added. After shaking for 1 minute, anaerobic culture was performed. After 24 hours, the supernatant was removed, 200 μL of PBS was added to wash the biofilm formed at the bottom of the 96-well plate, and the planktonic bacteria were removed and dried at room temperature. 200 μL of 0.1% crystal violet solution was added to each well, and shaking culture was performed for 20 minutes. 200 μL of PBS was added for washing 3 times, and the plate was dried at room temperature. 200 μL of 99% ethanol was added to each well, and shaking culture was performed for 1 hour. 100 μL of the supernatant was transferred to a new 96-well plate, and the absorbance was measured at a wavelength of 595 nm. A control group was set, and the control group was added with BHIA medium without test sample.

[0091] The formation inhibition rate (%) was calculated as follows: 1 - (absorbance of sample group) / (absorbance of control group) x 100%

[0092] Table 1: Results of bacteriostatic test of Streptococcus mutans

[0093]

[0094] The results of the bacteriostatic test of Streptococcus mutans according to the present application are shown in Table 1. When the content of bee oligopeptide reaches 1 ppm and 10 ppm, the test sample has good bacteriostatic effect on Streptococcus mutans.

[0095] Biofilm inhibition test based on Porphyromonas gingivalis

[0096] Porphyromonas gingivalis (P. gingivalis) has a preservation number of ATCC 33277, and the culture conditions are 80% N2 and 20% CO2, and culture at 37°C. Porphyromonas gingivalis Preparation of test sample: 0.01 g of sample was added to 10 mL of BHIA medium, and vortexed to disperse uniformly. Gradient dilution was performed to obtain test samples at corresponding test concentrations. The sample was synthetic bee oligopeptide with a purity of 99%.

[0097] Culture temperature: 23.1°C; humidity: 51%.

[0098] Preparation of Porphyromonas gingivalis suspension: Activated Porphyromonas gingivalis strain was taken, and 5 mL of PBS was added to prepare a bacterial suspension. The initial concentration of Porphyromonas gingivalis was adjusted to 1 x 10 8 CFU / mL, and it was prepared and used immediately.

[0099]

[0100] ​Inhibition test of P. gingivalis: In a 96-well plate, 180 μL of test sample at different concentrations was added, 20 μL of P. gingivalis suspension was added, and after shaking for 1 minute, anaerobic culture was performed. After 24 hours, the supernatant was removed, 200 μL of PBS was added to wash the biofilm formed at the bottom of the 96-well plate, and the planktonic bacteria were removed, and dried at room temperature. 200 μL of 0.1% crystal violet solution was added to each well, and shaking culture was performed for 20 minutes, 200 μL of PBS was added to wash 3 times, and dried at room temperature. 200 μL of 99% ethanol was added to each well, and shaking culture was performed for 1 hour, and 100 μL of supernatant was transferred to a new 96-well plate, and the absorbance was measured at a wavelength of 595 nm. A control group was set, and the control group was added with BHIA medium without test sample.

[0101] The inhibition rate (%) was calculated as 1 - (absorbance of sample group) / (absorbance of control group) x 100%.

[0102] Table 2: Results of biofilm inhibition test

[0103]

[0104] The results of the biofilm inhibition of P. gingivalis by the bee oligopeptide in the present application are shown in Table 2. When the content of bee oligopeptide reaches 1 ppm and 10 ppm, the test sample has good antibacterial effect on P. gingivalis.

[0105] Example 1: Preparation method of an oral care composition

[0106] The oral care composition comprises bee oligopeptide complex, regulator, and water. The bee oligopeptide complex comprises natural bee oligopeptide and synthetic bee oligopeptide, the natural bee oligopeptide has a purity of 60%, the synthetic bee oligopeptide has a purity of 99%, and the natural bee oligopeptide and the synthetic bee oligopeptide in the bee oligopeptide complex are mixed in a mass ratio of 1:9; the regulator is glycerol, PEG, and ethanol. The content of the bee oligopeptide complex in the oral care composition is 0.001 wt%, the content of glycerol in the oral care composition is 10 wt%, the PEG in the oral care composition is PEG300, the content of PEG300 in the oral care composition is 5 wt%, and the content of ethanol in the oral care composition is 15 wt%. The activity of water in the oral care composition is 0.87.

[0107] In this example, the oral care composition is prepared by mixing the bee oligopeptide complex, the regulator, and water.

[0108] Example 2: Preparation method of an oral care composition

[0109] The oral care composition comprises a bee oligopeptide complex, a regulator, an additive and water. The bee oligopeptide complex comprises natural bee oligopeptide and synthetic bee oligopeptide, the natural bee oligopeptide has a purity of 60%, the synthetic bee oligopeptide has a purity of 99%, and the natural bee oligopeptide and the synthetic bee oligopeptide are mixed in the bee oligopeptide complex at a mass ratio of 1:9; the regulator comprises glycerol, PEG and ethanol; and the additive comprises anhydrous citric acid, hydrogenated castor oil and essence. The content of the bee oligopeptide complex in the oral care composition is 0.001 wt%, the content of glycerol in the oral care composition is 10 wt%, the PEG in the oral care composition is PEG 300, the content of PEG 300 in the oral care composition is 5 wt%, the content of ethanol in the oral care composition is 15 wt%, the content of anhydrous citric acid in the oral care composition is 1.2 wt%, the content of hydrogenated castor oil in the oral care composition is 0.8 wt%, and the content of essence in the oral care composition is 0.2 wt%. The activity of water in the oral care composition is 0.85.

[0110] The oral care composition is prepared by mixing the bee oligopeptide complex, the regulator, the additive and water.

[0111] Example 3: A preparation method of an oral care composition

[0112] Preparation of mannuronic acid chloride: mannuronic acid is added to a solvent, and then dichlorosulfoxide is added at 5°C, and the reaction is carried out at 30°C for 4 hours. After the reaction is completed, the solvent and dichlorosulfoxide are removed by rotary evaporation to obtain mannuronic acid chloride. The solvent is dichloromethane, the use amount of mannuronic acid is 20 wt% of dichloromethane, and the use amount of dichlorosulfoxide is 70 wt% of mannuronic acid.

[0113] Preparation of mannuronic acid amide compound: an amino compound is added to an alkaline solution, and then mannuronic acid chloride is added, and the reaction is carried out at 30°C for 8 hours, and the pH is controlled at 10 during the reaction. After the reaction is completed, the pH is adjusted to 1, the precipitate is precipitated and filtered, washed and dried to obtain a mannuronic acid amide compound. The alkaline solution is a sodium hydroxide solution, the content of sodium hydroxide in the sodium hydroxide solution is 10 wt%, the amino compound is glycyl tyrosine, the use amount of the amino compound is 10 wt% of the sodium hydroxide solution, and the use amount of the mannuronic acid chloride is 200 wt% of the amino compound.

[0114] An oral care composition: a bee oligopeptide complex, a regulator, an additive and water. The bee oligopeptide complex comprises natural bee oligopeptide and synthetic bee oligopeptide, the natural bee oligopeptide has a purity of 60%, the synthetic bee oligopeptide has a purity of 99%, the natural bee oligopeptide and the synthetic bee oligopeptide in the bee oligopeptide complex are mixed in a mass ratio of 1:9; the regulator is glycerol, PEG and ethanol; the additive comprises anhydrous citric acid, hydrogenated castor oil, essence and a mannose amido compound. The content of the bee oligopeptide complex in the oral care composition is 0.001 wt%, the content of glycerol in the oral care composition is 10 wt%, the PEG in the oral care composition is PEG300, the content of PEG300 in the oral care composition is 5 wt%, the content of ethanol in the oral care composition is 15 wt%, the content of anhydrous citric acid in the oral care composition is 1.2 wt%, the content of hydrogenated castor oil in the oral care composition is 0.8 wt%, the content of essence in the oral care composition is 0.2 wt%, and the content of the mannose amido compound in the oral care composition is 0.07 wt%. The activity of water in the oral care composition is 0.84.

[0115] In this embodiment, an oral care composition is prepared by mixing a bee oligopeptide complex, a regulator, an additive and water.

[0116] Example 4: A preparation method of an oral care composition

[0117] In this embodiment, compared with Example 3, the content of the mannose amido compound in the oral care composition is 0.03 wt%. The activity of water in the oral care composition is 0.84.

[0118] Example 5: A preparation method of an oral care composition

[0119] In this embodiment, compared with Example 3, ascorbyl glycerin is additionally added to the additive. The content of ascorbyl glycerin in the oral care composition is 0.04 wt%. The activity of water in the oral care composition is 0.83.

[0120] Example 6: A preparation method of an oral care composition

[0121] In this embodiment, compared with Example 5, the content of ascorbyl glycerin in the oral care composition is 0.02 wt%. The activity of water in the oral care composition is 0.83.

[0122] Example 7: A preparation method of an oral care composition

[0123] In this embodiment, compared with Example 3, the content of the mannose amido compound in the oral care composition is 0.005 wt%. The activity of water in the oral care composition is 0.92.

[0124] The present application carries out infrared characterization on the mannose amido compound prepared in Example 3, and the results are shown in the following table, wherein the infrared absorption peak at 3408 cm-1 is that of hydroxyl, the infrared absorption peak at 2872 cm-1 is that of methylene, the infrared absorption peak at 1708 cm-1 is that of carbon-oxygen double bond, the infrared absorption peaks at 1628 cm-1 and 1492 cm-1 are those of benzene ring skeleton, and the infrared absorption peak at 1401 cm-1 is that of carbon-nitrogen bond, indicating that the mannose amido compound is obtained. Figure 1 -1 -1 -1 -1 -1 -1

[0125] The present application ages the oral care compositions prepared in each example at 40℃ and under a temperature condition of 75% for 2 weeks, and detects the content of bee oligopeptide in the oral care composition by HPLC. The loss rate of bee oligopeptide is calculated by comparing the content of bee oligopeptide before and after aging. According to the formula of the oral care composition of Example 2, the bee oligopeptide complex is replaced by natural bee oligopeptide, synthetic bee oligopeptide, bee oligopeptide complex 1 and bee oligopeptide complex 2 respectively, and the results are shown in Table 3.

[0126] Table 3 Stability test of bee oligopeptide

[0127]

[0128] In the present application, it is found that the stability of synthetic bee oligopeptide is the best, and there is basically no loss. The purity of bee oligopeptide in bee oligopeptide complex 1 is high, and the loss rate is small. The purity of bee oligopeptide in bee oligopeptide complex 2 is lower than 95%, and the loss rate is about 53% of that of natural bee oligopeptide. The loss rate of bee oligopeptide in bee oligopeptide complex 2 is lower than that of natural bee oligopeptide, indicating that when the purity of bee oligopeptide in bee oligopeptide complex is lower than 95%, the loss rate of bee oligopeptide has been greatly reduced.

[0129] The present application uses HFF-1 cells to test the cytotoxicity of the mannose amido compound prepared in Example 3. The HFF-1 cells are cultured in DMEM medium containing 0.12wt% mannose amido compound, and a control group is set. The control group is added with DMEM medium. The cell survival rate of the mannose amido compound group is compared with the control group as a reference. The cell survival rate of the HFF-1 cells cultured in DMEM medium containing 0.12wt% mannose amido compound is 99.6%, indicating that the mannose amido compound has basically no cytotoxicity.

[0130] The loss rates of bee oligopeptide of Examples 1-7 are as follows: Figure 2 ​​​​​​​The application is shown in the figure, wherein S1 is Example 1, S2 is Example 2, S3 is Example 3, S4 is Example 4, S5 is Example 5, S6 is Example 6, S7 is Example 7, the application prepares an oral care composition by mixing the bee oligopeptide complex, the regulator and water, the bee oligopeptide complex is prepared by compounding natural bee oligopeptide and synthetic bee oligopeptide in a certain proportion, and the use of the regulator can adjust the activity of water in the oral care composition and improve the stability of the bee oligopeptide. In the application, additives can also be added, and the use of the additives and the regulator together can also adjust the activity of water in the oral care composition, so that oral care compositions with different components can be prepared, and the use of the oral care composition can be expanded. In Example 1 of the application, the loss rate of bee oligopeptide in the obtained oral care composition is reduced by mixing specific bee oligopeptide complex, regulator and water, and after the additive in Example 2, the loss rate of bee oligopeptide in the obtained oral care composition is almost the same as that in Example 1. The additive used in Example 2 includes anhydrous citric acid, hydrogenated castor oil and essence, which indicates that the use of anhydrous citric acid, hydrogenated castor oil and essence has little effect on the reduction of the loss rate of bee oligopeptide after the use of the regulator. The additive in Examples 3-4 is added with a mannose amido compound, which is prepared from mannose chloride and an amino compound. The amino compound includes glycyl tyrosine, and the mannose chloride is prepared from mannose acid and dichlorosulfoxide. The use of the mannose amido compound can reduce the loss rate of bee oligopeptide, and the use amount of the mannose amido compound needs to be within a certain range. If the use amount of the mannose amido compound is too low, the loss rate of bee oligopeptide cannot be reduced. The additive in Examples 5-6 is also added with ascorbyl glyceride, and the use of ascorbyl glyceride and the mannose amido compound together can further reduce the loss rate of bee oligopeptide.

[0131] Example 8: A preparation method of an oral care composition

[0132] An oral care composition: a bee oligopeptide complex, a regulator, an additive, and water. The bee oligopeptide complex comprises natural bee oligopeptides and synthetic bee oligopeptides, the natural bee oligopeptides have a purity of 60%, the synthetic bee oligopeptides have a purity of 99%, the natural bee oligopeptides and the synthetic bee oligopeptides in the bee oligopeptide complex are mixed at a mass ratio of 1:9; the regulator is glycerol; the additive comprises xanthan gum, cellulose gum, sodium saccharin, sodium fluoride, hydroxybenzoic acid ester, hydrated silica, sodium lauryl sulfate, cocamidopropyl betaine, monohydrate citric acid, sodium hydroxide, and essence. The content of the bee oligopeptide complex in the oral care composition is 0.005 wt%, the content of the glycerol in the oral care composition is 2 wt%, the content of the xanthan gum in the oral care composition is 0.6 wt%, the content of the cellulose gum in the oral care composition is 0.6 wt%, the content of the sodium saccharin in the oral care composition is 0.35 wt%, the content of the sodium fluoride in the oral care composition is 0.24 wt%, the content of the hydroxybenzoic acid ester in the oral care composition is 0.1 wt%, the content of the hydrated silica in the oral care composition is 21 wt%, the content of the sodium lauryl sulfate in the oral care composition is 1.87 wt%, the content of the cocamidopropyl betaine in the oral care composition is 0.7 wt%, the content of the monohydrate citric acid in the oral care composition is 1.2 wt%, the content of the sodium hydroxide in the oral care composition is 0.04 wt%, and the content of the essence in the oral care composition is 1.2 wt%. The activity of water in the oral care composition is 0.98.

[0133] An oral care composition is prepared by mixing a bee oligopeptide complex, a regulator, an additive, and water.

[0134] Example 9: A preparation method of an oral care composition

[0135] Compared with Example 8, the difference of the present example lies in the regulator in the oral care composition. The regulator is glycerol and PEG, the content of the glycerol in the oral care composition is 5 wt%, the PEG is PEG300, and the content of the PEG300 in the oral care composition is 3 wt%. The activity of water in the oral care composition is 0.94.

[0136] Example 10: A preparation method of an oral care composition

[0137] Compared with Example 8, the difference of the present example lies in the regulator in the oral care composition. The regulator is glycerol, PEG, and sorbitol, the content of the glycerol in the oral care composition is 15 wt%, the PEG is PEG300, the content of the PEG300 in the oral care composition is 10 wt%, and the content of the sorbitol in the oral care composition is 15 wt%. The activity of water in the oral care composition is 0.85.

[0138] Example 11: A method of making an oral care composition

[0139] This example differs from Example 9 in that a mannose amido compound is also added to the additive. The amount of mannose amido compound in the oral care composition is 0.07 wt%. The activity of water in the oral care composition is 0.89.

[0140] Example 12: A method of making an oral care composition

[0141] This example differs from Example 9 in that a mannose amido compound is also added to the additive. The amount of mannose amido compound in the oral care composition is 0.03 wt%. The activity of water in the oral care composition is 0.89.

[0142] Example 13: A method of making an oral care composition

[0143] This example differs from Example 11 in that ascorbyl glycerin is also added to the additive. The amount of ascorbyl glycerin in the oral care composition is 0.04 wt%. The activity of water in the oral care composition is 0.88.

[0144] Example 14: A method of making an oral care composition

[0145] This example differs from Example 11 in that ascorbyl glycerin is also added to the additive. The amount of ascorbyl glycerin in the oral care composition is 0.02 wt%. The activity of water in the oral care composition is 0.88.

[0146] Example 15: A method of making an oral care composition

[0147] This example differs from Example 11 in that the amount of mannose amido compound in the oral care composition is 0.005 wt%. The activity of water in the oral care composition is 0.91.

[0148] Example 16: A method of making an oral care composition

[0149] An oral care composition: a bee oligopeptide complex, a regulator, an additive and water. The bee oligopeptide complex comprises natural bee oligopeptide and synthetic bee oligopeptide, the natural bee oligopeptide has a purity of 60%, the synthetic bee oligopeptide has a purity of 99%, the natural bee oligopeptide and the synthetic bee oligopeptide in the bee oligopeptide complex are mixed in a mass ratio of 1:9; the regulator is glycerol and D-panthenol; the additive comprises sucralose, a sugar lipid, monohydrate citric acid, sodium benzoate, essence and hydrogenated castor oil. The content of the bee oligopeptide complex in the oral care composition is 0.005wt%, the content of glycerol in the oral care composition is 2wt%, the content of D-panthenol in the oral care composition is 0.05wt%, the content of sucralose in the oral care composition is 0.6wt%, the content of the sugar lipid in the oral care composition is 0.6wt%, the content of monohydrate citric acid in the oral care composition is 0.35wt%, the content of sodium benzoate in the oral care composition is 0.24wt%, the content of essence in the oral care composition is 0.1wt%, and the content of hydrogenated castor oil in the oral care composition is 21wt%. The activity of water in the oral care composition is 0.97.

[0150] An oral care composition is prepared by mixing a bee oligopeptide complex, a regulator, an additive and water.

[0151] Example 17: A preparation method of an oral care composition

[0152] Compared with Example 16, the difference of the present example lies in the regulator in the oral care composition. The regulator is glycerol, PEG, propylene glycol, ethanol and D-panthenol, the content of glycerol in the oral care composition is 5wt%, the PEG is PEG300, the content of PEG300 in the oral care composition is 2wt%, the content of propylene glycol in the oral care composition is 1wt%, the content of ethanol in the oral care composition is 5wt%, and the content of D-panthenol in the oral care composition is 0.1wt%. The activity of water in the oral care composition is 0.94.

[0153] Example 18: A preparation method of an oral care composition

[0154] Compared with Example 16, the difference of the present example lies in the regulator in the oral care composition. The regulator is glycerol, PEG, propylene glycol, ethanol and D-panthenol, the content of glycerol in the oral care composition is 5wt%, the PEG is PEG300, the content of PEG300 in the oral care composition is 2wt%, the content of propylene glycol in the oral care composition is 1wt%, the content of ethanol in the oral care composition is 5wt%, and the content of D-panthenol in the oral care composition is 0.1wt%. The activity of water in the oral care composition is 0.94.

[0155] Example 19: A preparation method of an oral care composition

[0156] This example differs from example 17 in that a mannose amido compound is also added to the additive. The amount of mannose amido compound in the oral care composition is 0.07 wt%. The water activity of the oral care composition is 0.88.

[0157] Example 20: A method of making an oral care composition

[0158] This example differs from example 17 in that a mannose amido compound is also added to the additive. The amount of mannose amido compound in the oral care composition is 0.03 wt%. The water activity of the oral care composition is 0.88.

[0159] Example 21: A method of making an oral care composition

[0160] This example differs from example 19 in that ascorbyl glyceride is also added to the additive. The amount of ascorbyl glyceride in the oral care composition is 0.04 wt%. The water activity of the oral care composition is 0.87.

[0161] Example 22: A method of making an oral care composition

[0162] This example differs from example 19 in that ascorbyl glyceride is also added to the additive. The amount of ascorbyl glyceride in the oral care composition is 0.02 wt%. The water activity of the oral care composition is 0.87.

[0163] Example 23: A method of making an oral care composition

[0164] This example differs from example 19 in that the amount of mannose amido compound in the oral care composition is 0.005 wt%. The water activity of the oral care composition is 0.91.

[0165] Bee oligopeptide loss rate test

[0166] The oral care compositions prepared in each example were aged at 40 °C and 75% relative humidity for 30 days. The amount of bee oligopeptide in the oral care composition was determined using HPLC. The bee oligopeptide loss rate was calculated by comparing the amount of bee oligopeptide before and after aging.

[0167] The bee oligopeptide loss rates for examples 8-15 were as follows: Figure 3The loss rate of the bee oligopeptide in the oral care composition of Example 10 is about 2% through the use of different adjusting agents, the loss rate of the bee oligopeptide in the oral care composition of Example 9 is about 10.54%, and the use amount of the additives in Examples 8-10 is consistent, indicating that the use of the adjusting agent can reduce the loss rate of the bee oligopeptide, and the use of the additives in Examples 8-10 cannot significantly reduce the loss rate of the bee oligopeptide; the mannose amido compound is added to the additive in Examples 11-12 in the application, the mannose amido compound is made of mannose chloride and an amino compound, the amino compound includes glycyl tyrosine, the mannose chloride is made of mannuronic acid and thionyl chloride, the use of the mannose amido compound can reduce the loss rate of the bee oligopeptide, and the use amount of the mannose amido compound needs to be within a certain range, if the use amount of the mannose amido compound is too low, the loss rate of the bee oligopeptide cannot be basically reduced; the ascorbyl glycoside is further added to the additive in Examples 13-14, and the common use of the ascorbyl glycoside and the mannose amido compound can further reduce the loss rate of the bee oligopeptide.

[0168] The loss rates of the bee oligopeptides in Examples 16-23 are as follows: Figure 4The loss rate of the bee oligopeptide in the oral care composition of Example 18 is about 1.33%, the loss rate of the bee oligopeptide in the oral care composition of Example 9 is about 15.52%, and the use amount of the additive in Examples 16-18 is consistent, indicating that the use of the adjusting agent can reduce the loss rate of the bee oligopeptide, and the use of the additive in Examples 16-18 cannot significantly reduce the loss rate of the bee oligopeptide; the additive in Examples 19-20 in the application adds a mannose amido compound, the mannose amido compound is made of mannose chloride and an amino compound, the amino compound includes glycyl tyrosine, the mannose chloride is made of mannuronic acid and dichlorosulfoxide, the use of the mannose amido compound can reduce the loss rate of the bee oligopeptide, and the use amount of the mannose amido compound needs to be in a certain range, if the use amount of the mannose amido compound is too low, the loss rate of the bee oligopeptide cannot be basically reduced; the additive in Examples 21-22 further adds ascorbyl glyceride, and the common use of the ascorbyl glyceride and the mannose amido compound can further reduce the loss rate of the bee oligopeptide.

[0169] The above-described embodiments and / or implementations are merely used to illustrate the preferred embodiments and / or implementations of the present application, and not intended to limit the embodiments of the present application in any form, and any person skilled in the art can make some changes or modifications as other equivalent embodiments without departing from the scope of the technical means disclosed in the present application, and the technical or embodiments substantially the same as the present application should be considered.

[0170] The principles and implementations of the present application are described by using specific examples herein, and the above example descriptions are only used to help understand the method and core idea of the present application. The above descriptions are only preferred embodiments of the present application, and it should be pointed out that due to the limited expression, there are infinite specific structures, and for ordinary skilled persons in the technical field, some improvements, refinements or changes can be made without departing from the principles of the present application, and the above technical features can be combined in an appropriate manner; these improvements, refinements, changes or combinations, or the direct application of the inventive concept and technical solution to other occasions without improvement, should be considered as the protection scope of the present application.

Claims

1. An oral care composition comprising: The bee oligopeptide complex is mixed with a modulator, the purity of the bee oligopeptide in the bee oligopeptide complex is more than 95%, and the mass ratio of the bee oligopeptide complex to the modulator is 0.02-1:10-8140; The modulator comprises at least one of glycerol, PEG, ethanol, sorbitol, propylene glycol, and D-panthenol; The mass ratio of the bee oligopeptide complex to glycerol is 0.02-1:300-2000; or, The mass ratio of the bee oligopeptide complex to PEG is 0.02-1:200-1500; or, The mass ratio of the bee oligopeptide complex to ethanol is 0.02-1:1000-2000; or, The mass ratio of the bee oligopeptide complex to sorbitol is 0.02-1:1000-2000; or, The mass ratio of the bee oligopeptide complex to propylene glycol is 0.02-1:100-600; or, The mass ratio of the bee oligopeptide complex to D-panthenol is 0.02-1:10-40. The oral care composition further comprises an additive, the additive comprises a mannuronyl amino compound, the mannuronyl amino compound comprises a mannuronyl group and an amino compound group; in the preparation of the mannuronyl amino compound, first, a mannuronyl chloride is prepared by reacting mannuronic acid and dichloro sulfoxide, and then the mannuronyl chloride is reacted with an amino compound to obtain the mannuronyl amino compound; the content of the mannuronyl amino compound in the oral care composition is 0.02-0.12 wt%.

2. An oral care composition according to Claim 1 characterised in that: The additive comprises at least one of anhydrous citric acid, hydrogenated castor oil, essence, xanthan gum, cellulose gum, sodium saccharin, sodium fluoride, hydroxybenzoic acid ester, hydrated silica, sodium lauryl sulfate, cocamidopropyl betaine, sucralose, sugar lipid surfactant, monohydrate citric acid, sodium benzoate, PEG40, and inorganic salt.

3. An oral care composition according to Claim 2 characterised in that: The inorganic salt comprises at least one of sodium chloride, sodium hydroxide, sodium carbonate, sodium bicarbonate, potassium chloride, potassium hydroxide, potassium carbonate, and potassium bicarbonate.

4. The oral care composition according to claim 2, wherein: The mass ratio of the bee oligopeptide complex to xanthan gum is 0.02-1:30-120; or, The mass ratio of the bee oligopeptide complex to cellulose gum is 0.02-1:30-120; or, The mass ratio of the bee oligopeptide complex to sodium saccharin is 0.02-1:20-70; or, The mass ratio of the bee oligopeptide complex to sodium fluoride is 0.02-1:10-50; or, The mass ratio of the bee oligopeptide complex to hydroxybenzoic acid ester is 0.02-1:5-20; or, The mass ratio of the bee oligopeptide complex to hydrated silica is 0.02-1:1000-3000; or, The mass ratio of the bee oligopeptide complex to sodium lauryl sulfate is 0.02-1:100-400; or, The mass ratio of the bee oligopeptide complex to cocamidopropyl betaine is 0.02-1:30-140; or, The mass ratio of the bee oligopeptide complex to sucralose is 0.02-1:0.2-0.8; or, The mass ratio of the bee oligopeptide complex to sugar lipid is 0.02-1:7-30; or, the mass ratio of the bee oligopeptide complex and citric acid monohydrate is 0.02-1:10-40; or, the mass ratio of the bee oligopeptide complex and sodium benzoate is 0.02-1:25-100; or, the mass ratio of the bee oligopeptide complex and inorganic salt is 0.02-1:2-8; or, the mass ratio of the bee oligopeptide complex and anhydrous citric acid is 0.02-1:60-240; or, the mass ratio of the bee oligopeptide complex and hydrogenated castor oil is 0.02-1:40-160; or, the mass ratio of the bee oligopeptide complex and essence is 0.02-1:10-40.

5. Use of the oral care composition of any one of claims 1-4 in the preparation of an oral care product.

6. Use according to claim 5, characterized in that: The oral care product includes mouthwash and / or toothpaste.

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