Composition for oral care as well as preparation method and application thereof

Abstract

The invention relates to the technical field of oral care, in particular to a composition for oral care and a preparation method and application thereof. The composition is prepared from the following components in parts by mass: 0.5 to 8.0 parts of recombinant human albumin and 0.5 to 8.0 parts of calyx seu fructus physalis extract. The mass part of the calyx seu fructus physalis extract is 0.5-2.0 parts. The recombinant human albumin and the calyx seu fructus physalis extract have a synergistic effect in antibacterial and antioxidant aspects, can be used for oral care or oral inflammation treatment, particularly has important effects in oral foreign matter removal and tooth cleaning, has an obvious antibacterial effect, and expands the application field of the recombinant human albumin.

Description

Technical Field

[0001] The present invention relates to the technical field of oral care, and in particular to a composition for oral care, a preparation method and application thereof. Background Art

[0002] With the development of society, people's material and cultural life is becoming more and more abundant, and the requirements for oral health are also getting higher and higher. Due to the large amount of tobacco, tea, coffee, colored foods and carbonated drinks in daily life, some of them always remain and deposit on the surface of teeth when passing through the mouth. After a long period of accumulation, the teeth will turn yellow and black, affecting people's normal social life.

[0003] Toothpaste, as a basic product for oral cleaning, has become a necessity in people's daily life. The ingredients of existing toothpaste products are mostly based on chemical synthetic substances. Although these substances have certain effects in cleaning the oral cavity, long-term use may have a negative impact on the oral environment, such as destroying the oral ecological balance and causing tooth sensitivity. As people pay more attention to oral health, their expectations for toothpaste are no longer limited to basic cleaning functions, but they pay more attention to its functionality, safety and naturalness. Plant extracts are gradually gaining attention in the field of oral care due to their natural and safe characteristics.

[0004] In order to make toothpaste more effective, many inventions have tried to add various drugs to toothpaste, but these drugs often have toxic effects on the human body, especially after long-term use of medicated toothpaste, the side effects are large. Therefore, it is urgent to develop a toothpaste that has no side effects on the human body and has strong efficacy.

[0005] Human serum albumin (HSA) is a monomeric protein produced by the liver. Mature human serum albumin contains 585 amino acid residues and has a molecular weight of 66KD. Albumin has abundant free thiol groups and antioxidant properties, and can scavenge free radicals (such as hypochlorous acid (HOCl) and peroxynitrite (ONOOH)). Due to its large molecular weight and many active groups, it can bind to teeth and microorganisms in the mouth. These advantages make albumin very suitable as a raw material for toothpaste. So far, there has been no research on the use of albumin to prepare oral care products. Summary of the invention

[0006] To solve the above problems, the first aspect of the present invention provides a composition for oral care, which comprises recombinant human albumin and Lantern Festival Chinese Phalaenopsis extract, wherein the mass fractions of the components are as follows: the mass fraction of recombinant human albumin is 0.5~8.0 parts; the mass fraction of Lantern Festival Chinese Phalaenopsis extract is 0.5~2.0 parts.

[0007] Furthermore, the composition also includes additives acceptable in the field of oral care products and medicines.

[0008] Furthermore, the composition also includes at least one of abrasives, moisturizers, thickeners, foaming agents, anti-sensitivity agents, colorants, flavoring agents, preservatives, whitening agents, fluorides, surfactants, buffers, lubricants, fragrances, pH regulators, and emulsifiers.

[0009] Further, the abrasive comprises at least one of calcium carbonate, silica, and silicon dioxide; the moisturizer comprises at least one of glycerin, sorbitol, propylene glycol, sodium phytate, and polyethylene glycol; the thickener comprises at least one of sodium carboxymethyl cellulose and xanthan gum; the foaming agent comprises at least one of sodium lauroyl amino acid, cocamidopropyl betaine, trideceth-9, polyglyceryl laurate, PEG-20 hydrogenated castor oil, and lauryl glucoside; the anti-sensitizer comprises at least one of potassium nitrate and strontium chloride; the colorant comprises at least one of chlorophyll (green), curcumin (yellow), and beetroot red (red); the flavoring comprises at least one of mint, sucralose, sodium saccharin, and edible flavors; the preservative comprises at least one of sodium benzoate and parabens; the whitening agent comprises The invention relates to a novel surfactant comprising at least one of hydrogen peroxide, sodium phytate, pentasodium tripolyphosphate, sodium hexametaphosphate, sodium pyrophosphate, tetrasodium pyrophosphate, tetrapotassium pyrophosphate and disodium dihydrogen pyrophosphate; the fluoride comprises at least one of sodium fluoride and sodium monofluorophosphate; the surfactant comprises at least one of carbomer, sodium lauryl sulfate, sodium lauroyl sarcosine, sodium lauryl sulfate, sodium lauroyl glutamate and cocoyl glucoside; the buffer comprises at least one of sodium hydrogen phosphate, sodium dihydrogen phosphate, sodium silicate, potassium silicate and sodium bicarbonate; the lubricant comprises at least one of vegetable glycerin and xylitol; the pH adjuster comprises at least one of sodium citrate; the emulsifier comprises at least one of glycerol monostearate, sorbitan laurate, sorbitan palmitate, sorbitan stearate, polysorbate 20, polysorbate 40 and polysorbate 80.

[0010] It should be noted that the present invention does not strictly limit the composition and weight fraction of additives acceptable in the field of cosmetics. All additives that are harmless to the human body and have the functions of abrasives, moisturizers, thickeners, foaming agents, anti-sensitizers, colorants, flavorings, preservatives, whitening agents, fluorides, surfactants, buffers, lubricants, fragrances, pH adjusters, emulsifiers, or one or more additives selected according to the dosage form of the product, or the weight fraction of the additives is routinely adjusted according to actual needs, are all protected by the present invention.

[0011] The second aspect of the present invention provides the use of the composition in at least one of the following aspects: A1. Application in the preparation of antibacterial products; A2. Application in the preparation of antioxidant products; A3. Application in the preparation of products for inhibiting oral inflammation; A4. Application in the preparation of products for treating and / or preventing dental caries.

[0012] Furthermore, the product includes at least one of an oral care product or a medicine.

[0013] Furthermore, the product includes at least one of toothpaste and mouthwash; further, the oral medicine includes medicine for treating oral inflammation.

[0014] Furthermore, the oral inflammation described in Application A3 includes at least one of gingivitis, periodontitis, oral ulcers, oral mucositis, or oral inflammation caused by Staphylococcus aureus, Candida albicans, Prevotella intermedia, Porphyromonas gingivalis, Steinerella forsythia, Fusobacterium nucleatum, Actinobacillus actinomycetemcomitans, and Streptococcus mutans.

[0015] The third aspect of the present invention provides a toothpaste containing recombinant human albumin, the toothpaste comprising: 0.5 to 8.0 parts by mass of recombinant human albumin; 0.5 to 2.0 parts by mass of Lantern Festival extract; 21.1 to 50.4 parts by mass of moisturizer; 10 to 25 parts by mass of abrasive; 1.05 to 7.2 parts by mass of surfactant; 0.42 to 1.2 parts by mass of flavoring agent; 0.05 to 0.3 parts by mass of preservative; 0.02 to 0.20 parts by mass of emulsifier; and 0.4 to 3.5 parts by mass of thickener.

[0016] Furthermore, the moisturizer includes at least one of sorbitol, propylene glycol, and sodium phytate; the abrasive includes silicon dioxide; the surfactant includes at least one of carbomer, sodium lauryl sulfate, and sodium lauroyl sarcosinate; the thickener includes at least one of sodium carboxymethyl cellulose and xanthan gum; the emulsifier includes polysorbate 80; the flavoring agent includes at least one of sucralose, sodium saccharin, and edible flavors; and the preservative includes sodium benzoate.

[0017] Preferably, the components and mass fractions of the toothpaste are as follows: 0.5-8.0 mass fraction of recombinant human albumin; 0.5-2.0 mass fraction of Lantern Festival extract; 20-45 mass fraction of sorbitol; 1.0-5.0 mass fraction of propylene glycol; 10-25 mass fraction of silicon dioxide; 0.3-2.0 mass fraction of sodium carboxymethyl cellulose; 0.1-1.5 mass fraction of xanthan gum; 0.05-1.2 mass fraction of carbomer; and 0.5-1.0 mass fraction of sucrose. The mass fraction of sugar is 0.01-0.10 parts; the mass fraction of saccharin sodium is 0.01-0.10 parts; the mass fraction of sodium lauryl sulfate is 0.5-3.0 parts; the mass fraction of sodium lauroyl sarcosinate is 0.5-3.0 parts; the mass fraction of sodium benzoate is 0.05-0.3 parts; the mass fraction of edible flavor is 0.4-1.0 parts; the mass fraction of polysorbate 80 is 0.02-0.20 parts; the mass fraction of sodium phytate is 0.10-0.40 parts; and an appropriate amount of deionized water is used for dissolving.

[0018] A fourth aspect of the present invention provides a method for preparing the composition, the composition comprising the following steps: S1: Preparation of Lantern Festival extract: crush Lantern Festival, pass through a 10-mesh sieve to obtain powder, soak it in 70%~85% (v / v) ethanol at a material-liquid mass ratio of 1:(3~8), reflux extraction in a water bath at 40~100°C, distill to remove ethanol until there is no alcohol taste, add deionized water to dissolve and mix the Lantern Festival extract, spray dry the mixed solution to obtain the Lantern Festival extract; S2: Dissolve the Lantern Festival extract in a certain amount by mass into a recombinant human albumin aqueous solution or a salt buffer, and use a high-pressure homogenizer at 300-1000 bar for high-pressure homogenization for 25-35 min at a temperature of 25-50°C to obtain a uniform solution of the composition. Preferably, the recombinant human albumin salt buffer comprises at least one of recombinant human albumin phosphate buffer, recombinant human albumin citrate buffer, and recombinant human albumin acetate buffer. Preferably, the recombinant human albumin buffered saline solution is a recombinant human albumin PBS buffer.

[0019] Furthermore, the spray drying conditions in step S1 are: air inlet temperature: 120-160° C., atomization pressure: 0.1-0.5 MPa, and feed rate: 300-700 ml / h.

[0020] Further, the ethanol concentration in step S1 is 70%, 73%, 75%, 78%, 80%, 83%, 85% or any value therebetween; the mass ratio of the Chinese lantern to ethanol is 1:3, 1:3.5, 1:4, 1:4.5, 1:5, 1:5.5, 1:6, 1:6.5, 1:7, 1:7.5, 1:8 or any value therebetween. Further, the time for soaking the Chinese lantern extract in ethanol is 4 to 8 hours.

[0021] Furthermore, the extraction time of the water bath extraction step is 15 to 25 minutes; further, the water bath extraction step can be repeated 2 to 5 times, further, the water bath temperature extraction step can be repeated 2, 3, 4, 5 times, preferably, the step is repeated 3 times; further, the filtrates of the repeatedly extracted Lantern Festival extract need to be combined and then distilled.

[0022] Furthermore, the method comprises the step of adding additives acceptable in the field of oral care products and medicines to the composition to prepare the oral care product or medicine.

[0023] In a specific embodiment of the present invention, a method for preparing recombinant human albumin toothpaste using the composition is provided, the method comprising the following steps: (1) Preparation of Lantern Festival extract: crush the Lantern Festival used in the formula, pass it through a 10-mesh sieve to obtain a powder, soak it in 70%-85% ethanol at a solid-liquid ratio of 1:(3-8) for 4-8 hours, reflux extract it in a water bath at 40-100°C for 15-25 minutes, extract it three times, combine the filtrate, distill off the ethanol until there is no alcohol taste, suspend 0.5-2.0 parts of Lantern Festival extract in deionized water, mix the above solutions evenly, and spray dry the mixed solution to obtain Lantern Festival extract; (2) Inlet air temperature for spray drying: 120~160℃, atomization pressure: 0.1~0.5Mpa, feed rate: 300~700ml / h; (3) Add the Lantern Festival extract, sodium phytate, sodium benzoate, edible flavor, sucralose, sodium saccharin, and polysorbate 80 according to weight proportions into deionized water until completely dissolved, as phase A; take weight proportions of xanthan gum, carbomer, sodium carboxymethyl cellulose, sodium lauryl sulfate, sodium lauroyl sarcosinate, and mixed powder, add them into sorbitol and propylene glycol, place them on a magnetic stirrer and stir at a speed of 400-1000 rpm and a temperature of 50-85°C to make the adhesive Evenly disperse in the moisturizer and foaming agent as phase B, add phase A to phase B, the adhesive rapidly swells into a colloid when in contact with water, continue stirring until it is evenly mixed, transparent and free of powdery particles, store and age for 2-6 hours (preferably 4 hours), and further stir until the obtained colloid is uniform and transparent as phase C; add silicon dioxide to phase C at a speed of 100-400 rpm, mix it thoroughly, and stir at a speed of 100-400 rpm for 30-60 minutes as phase D. After mixing phase D with the recombinant human albumin solution, use a high-pressure homogenizer at 300-1000 bar, high-pressure homogenization for 30 minutes, and a temperature of 25-50°C to obtain a uniform paste; (4) The uniform paste obtained after high-pressure homogenization is placed in a -4°C refrigerator and aged for 24 hours to obtain the recombinant human albumin toothpaste.

[0024] The beneficial effects of the present invention include but are not limited to: 1. In the composition of the present invention, recombinant human albumin and Lantern Festival extract work synergistically to clean the oral cavity from the two aspects of antibacterial and anti-oxidation, can be used to prepare products for inhibiting oral inflammation, and can also effectively improve the situation of dental caries. In addition, the composition of the present invention has simple ingredients and good biocompatibility, and can be widely used in the preparation of oral care products or oral medicines.

[0025] 2. The preparation method of the composition of the present invention has a simple production process and has a high application prospect. DETAILED DESCRIPTION

[0026] The present invention is described in detail below with reference to examples, but the present invention is not limited to these examples. Unless otherwise specified, the raw materials and catalysts in the examples of the present invention are purchased through commercial channels.

[0027] Definition: Recombinant human albumin refers to albumin prepared using recombinant DNA technology for treatment and prevention.

[0028] Source of materials: Recombinant human albumin: produced by Tonghua Anrui Biopharmaceutical Co., Ltd., product number: ART101S; The brocade lanterns were purchased from the Anguo Chinese Medicine Market in Hebei Province.

[0029] equipment: Water bath (HH.S21-8, Shanghai Boxun Medical Biological Instrument Co., Ltd.); Multifunctional fluorescence microplate reader (SPMax 3500FL, Shanghai Shanpu Biotechnology Co., Ltd.); Analytical balance (BSA224S, Sartorius Scientific Instruments Beijing Co., Ltd.); pH meter (PB-10, Sartorius Scientific Instruments Beijing Co., Ltd.) Unless otherwise specified, other reagents and instruments used were commercially available.

[0030] Example 1 Preparation of a composition containing recombinant human albumin comprising the following components in parts by weight: The effective ingredients are shown in Table 1.

[0031] Table 1 Compositions of various compositions containing recombinant human albumin

[0032] The specific steps for preparing a composition containing recombinant human albumin are as follows: The second aspect of the present invention provides a method for preparing a composition containing recombinant human albumin, comprising the following steps: (1) The Lantern Festival extract is prepared according to the following method: crush the Lantern Festival used in the formula, pass it through a 10-mesh sieve to obtain a powder, soak it in 75% ethanol at a solid-liquid ratio of 1:5 for 6 hours, reflux extract for 15 minutes at a water bath temperature, extract it 3 times, combine the filtrate, distill off the ethanol until there is no alcohol taste, suspend the Lantern Festival extract in deionized water, mix the above solutions evenly, and spray dry the mixed solution to obtain; (2) Inlet air temperature for spray drying: 120~160℃, atomization pressure: 0.1~0.5Mpa, feed rate: 300~700ml / h; (3) Dissolve the Lantern Festival extract in a recombinant human albumin aqueous solution or salt buffer according to the mass fraction, and use a high-pressure homogenizer at 300-1000 bar for 30 minutes at a temperature of 25-50°C to obtain a uniform recombinant human albumin composition solution.

[0033] Test Example 1 Antibacterial Test The antibacterial effect of the composition was examined with reference to QB / T2738-2012 "Evaluation Methods for Antibacterial and Antimicrobial Effects of Daily Chemical Products" "7.5 Test Methods for Antibacterial Effects of Antibacterial Daily Chemical Products (Inhibitory Ring Method)".

[0034] Take the recombinant human albumin composition solution of groups 1 to 13 in Example 1, and use a pipette to draw 50 μl of bacterial suspension (1×10 4 ~ 9×10 4CFU / ml) were placed on the surface of the corresponding nutrient agar medium plate, evenly spread 3 times, covered the plate, and dried at room temperature for 5 minutes. Take sterile dry filter paper pieces, add 20μl of the original solution of the composition to each piece as the test sample, and take 20μl of sterile distilled water as the control sample. Put 3 test pieces on each plate, 1 piece for the photo, cover the plate, seal it with sealing film, place it in a constant temperature incubator at a certain temperature for 16~18h, observe it, use a vernier caliper to accurately measure the diameter of the inhibition zone, and take the average value. When the test bacteria is Staphylococcus aureus, the culture dish is placed in a constant temperature box at 37°C for 16-18 hours; when the test bacteria is Candida albicans, the culture dish is placed in a constant temperature box at 28°C for 16-18 hours; when the test bacteria is Porphyromonas gingivalis, the culture dish is placed in a constant temperature box at 37°C for 72-96 hours; when the test bacteria is Prevotella intermedia, the culture dish is placed in a constant temperature box at 37°C for 24-48 hours; when the test bacteria is T. forsythiae, the culture dish is placed in a constant temperature box at 37°C for 24-48 hours. The dish was placed in a 25°C incubator for 24-48 hours; when the test bacteria was Fusobacterium nucleatum, the dish was placed in a 37°C incubator for 72-96 hours; when the test bacteria was Actinobacillus actinomycetemcomitans, the dish was placed in a 37°C incubator for 24-48 hours; when the test bacteria was Streptococcus mutans, the dish was placed in a 37°C incubator for 18-24 hours; when the test bacteria was Lactobacillus, the dish was placed in a 37°C incubator for 16-24 hours. The test was repeated 3 times. If the diameter of the antibacterial ring is greater than 7mm, the product has an antibacterial effect, otherwise the product has no antibacterial effect. The control group (Group 14) was sterile deionized water. The results of the antibacterial ring determination are shown in Table 2.

[0035] Table 2 Results of inhibition zone determination of antibacterial experiments of various compositions containing recombinant human albumin

[0036] Note: “——” No inhibition zone was detected.

[0037] It can be concluded from Table 2 that the recombinant human albumin composition showed good antibacterial activity against Staphylococcus aureus, Candida albicans, Prevotella intermedia, Porphyromonas gingivalis, Steinerella forsythia, Fusobacterium nucleatum, Actinobacillus actinomycetemcomitans and Streptococcus mutans, and the average diameter of the inhibition zone was greater than 7 mm, indicating that the prepared recombinant human albumin composition was moderately sensitive to the above pathogens. The groups (6-9) prepared according to the optimal ratio of plant-based and animal-based albumin were far lower than recombinant human albumin in terms of antibacterial effect, indicating that the prepared recombinant human albumin had the most significant antibacterial effect. The recombinant human albumin compositions prepared in Group 4 and Group 3 had the best antibacterial activity; the antibacterial effect gradually increased with the increase in the mass fractions of recombinant human albumin and Lanternaria officinalis (Group 1 and Group 2); but after the mass fractions of recombinant human albumin and Lanternaria officinalis increased to the optimal ratio, the antibacterial effect began to decrease (Group 5 and Group 13); the antibacterial effect of no recombinant human albumin (Group 10) and no Lanternaria officinalis component (Group 11) was much lower than the antibacterial effect of adding both at the same time. This shows that in the recombinant human albumin composition prepared by this method, recombinant human albumin and Lanternaria officinalis are functional ingredients.

[0038] Test Example 2 Anti-caries Test The compositions prepared by group 4, group 3, and group 2 were taken; 90 subjects were selected, and the inclusion criteria of the subjects were as follows: the subjects were in good general condition; the number of natural teeth in the mouth was ≥15; accompanied by chronic gingivitis; and they had the habit of brushing their teeth in the morning and evening before the test. Excluded were those with caries, pulp disease, or apical lesions and untreated; those with damaged oral mucosa; those with denture repair, orthodontic appliances, and poor restorations in the mouth; those who used antibiotics or tetracycline drugs during the trial; those who were allergic to oral cleaning products; and those with infectious diseases such as diabetes or hepatitis B. The subjects were randomly divided into 3 groups, 30 in each group. Group 1 used the composition prepared by group 4, group 2 used the composition prepared by group 3, and group 3 used the composition prepared by group 2. There was no significant difference in general data such as gender and age among the subjects in each group. The subjects in each group brushed their teeth once in the morning and evening, 2 minutes / time, for 6 consecutive weeks. The subjects in each group could not change other oral cleaning products during the trial, and the effect was evaluated after 6 weeks of use. Gum index score: (1) 0 points: healthy gums; (2) 1 point: no inflammation of the gums, but slight changes in gum color and edema, no bleeding after probing; (2) 2 points: mild inflammation of the gums, slight changes in gum color and edema, no bleeding after probing; (3) 3 points: moderate inflammation of the gums, red gums, edema and shiny, bleeding after probing; (4) 4 points: severe inflammation of the gums, obvious redness, swelling and ulceration of the gums, and a tendency to spontaneous bleeding. The test results are shown in Table 3.

[0039] Table 3 The therapeutic effects of various compositions containing recombinant human albumin on gingivitis (scores)

[0040] According to the data in Table 3, the composition prepared by the method disclosed in this patent can relieve the symptoms of patients with severe gingival inflammation and moderate gingival inflammation, and has a therapeutic and repairing effect on patients with gingival disease, indicating that the composition prepared by this patent has a good anti-inflammatory effect.

[0041] Test Example 3 Oral ulcer healing experiment Experimental content: A total of 60 clean-grade SD rats (weighing 250-300g) were modeled by intraperitoneal injection of anesthesia phenol cauterization. Criteria for successful modeling: a circular or oval ulcer with a diameter of about 5mm is formed in the cauterized area, surrounded by redness and swelling, the mucosal surface is rough and uneven, there is a gray or yellow pseudomembrane, the activity, food intake, and water intake are reduced, and the stool is thin. On the second day after modeling, 60 male SD rats with oral ulcers that were successfully modeled were taken and randomly divided into 10 groups, 6 in each group, namely the model group, experimental groups 1-8 and the positive control group. Among them, the model group did not apply any drugs, and the experimental groups 1-8 were respectively applied with the recombinant human albumin compositions prepared by groups 3, 4, 7, 9, 10, 11, 12, and 13 in Example 1, and the positive control group was applied with Bingpengsan (commercially available). Use a sterilized cotton swab to apply the corresponding medicine to the oral ulcer. Take the composition prepared by Group 3, Group 4, Group 7, Group 9-12 and Bingpengsan powder to completely cover the wound surface. Apply it once in the morning and evening, with an interval of 8 hours between administrations. Treat continuously for 7 days, and take the complete healing of the ulcer as the end point. Observe the changes in the oral mucosa every day, measure the area of ​​the mucosal ulcer without edema, and observe the inflammation and congestion. Detection time points: before application after modeling and 8 hours after the completion of administration. Use a vernier caliper to measure the diameter of the ulcer and calculate the ulcer area. At the same time, record the water intake and food intake of each animal per unit time. Evaluation index: record the ulcer area on the 7th day after application: 1mm 2 For healing, ≥1mm 2 The model group is a blank control group without any drug. The results of the oral ulcer effect test on rat oral ulcers in the model group, positive control group and experimental groups 1 to 8 are shown in Table 4.

[0042] Table 4 The therapeutic effects of various compositions containing recombinant human albumin on oral ulcers

[0043] Note: Compared with the model group, P<0.05, * has significant difference.

[0044] As shown in Table 4, compared with the model group, both the drug-administered group and the commercially available group played a role in oral healing. The effects of the single component group (experimental group 5-6) and the non-optimal formula group (experimental group 7-8) were much lower than those of the optimal ratio group of recombinant human albumin, and the wounds could not heal until at least 7 days later. The optimal formula group (experimental group 1-2) showed a significant effect of promoting the healing of oral ulcers in rats in 4 days, and its effect was better than that of the positive control group (Bingpengsan group). In experimental groups 3-4, the effects of plant-derived and animal-derived albumin on healing oral ulcer wounds were not as good as those of the positive control group, and the healing effect was poor, which further showed that recombinant human albumin was more conducive to the treatment of oral ulcers.

[0045] Test Example 4 Antioxidant Experiment-DPPH Scavenging Ability Determination: Preparation of DPPH solution: Accurately weigh 20 mg of DPPH, dissolve it in anhydrous ethanol and make up to 500 ml in a brown volumetric flask to obtain a DPPH solution with a mass fraction of 0.004%. Store it away from light for later use.

[0046] Vitamin C was selected as the positive control. The experimental groups 1 to 8 (the compositions prepared by groups 3, 4, 7, 9 to 13 in Example 1 were respectively administered) and the positive control solution were prepared with a starting concentration of 1 mg / mL, and the concentration range was 0.002 to 1 mg / mL. The 96-well plate was divided into a sample group and a control group, with 3 replicate wells in each group. A blank control was set for both the sample group and the control group. The blank group was anhydrous ethanol, which was added to the DPPH solution in an equal volume of 1:1. The 96-well plate was reacted in the dark at room temperature for 30 minutes, shaken and evenly mixed, and the absorbance was measured at a wavelength of 517 nm. The half inhibitory concentration (IC 50 ) value (μg / ml). IC 50 The IC value is an important indicator to measure the strength of antioxidant activity, which indicates the concentration required for the antioxidant capacity to achieve half of the clearance effect. 50 The smaller the value, the better the free radical scavenging effect. The data results are shown in Table 5. The calculation formula for the free radical scavenging rate is as follows: Free radical scavenging rate (%) = ( )×100% Wherein: A1 is the measured OD value of the sample group, A2 is the measured OD value of the sample control group, and A0 is the measured OD value of the blank group.

[0047] Table 5 Results of DPPH antioxidant treatment with various compositions containing recombinant human albumin

[0048] Note: Compared with vitamin C, P<0.05, * has significant difference.

[0049] From the data results in Table 5, it can be seen that experimental groups 1~2 and 5~8 all have DPPH scavenging activity, among which experimental groups 1~2 and 5~8 are better than vitamin C as a positive control.

[0050] IC of DPPH scavenging activity of plant and animal albumin groups (experimental groups 3 and 4) 50 The value is the highest, indicating that albumin from plant and animal sources has the lowest antioxidant capacity.

[0051] Compared with the non-optimal ratio groups (experimental groups 7-8), experimental groups 5~6 (group 10 without the addition of recombinant human albumin, group 11 without the addition of Lantern Festival extract) had weaker DPPH scavenging ability, which further indicated that recombinant human albumin and Lantern Festival extract must be used in a specific ratio to play a synergistic role.

[0052] Test Example 5 Anti-inflammatory Experiment The effect of xylene on the swelling of the auricle of mice was tested to study its anti-inflammatory effect. Deionized water was used as the negative control group, dexamethasone acetate ointment was used as the positive control group, and the compositions obtained from Group 3, Group 4, Group 7, Group 9, Group 10, Group 11, Group 12, and Group 13 were used as test groups 1 to 8. 100 SPF female mice were randomly divided into 10 groups according to their body weight, with 10 mice per group. They were respectively the negative control group, the positive control group, and the experimental groups 1 to 8 (the compositions prepared from Groups 3, 4, 7, 9 to 13 in Example 1 were administered respectively).

[0053] Xylene was evenly applied to the inner and outer surfaces of the right auricle of each mouse to cause inflammation, with a dose of 100 μl / mouse (50 μl / mouse on one side), and the left ear was not treated as a blank control group. 10 minutes after xylene induced inflammation, the mice were first smeared with deionized water (0.1 ml / mouse) and dexamethasone acetate ointment (0.1 g / mouse) according to the corresponding groups, and the experimental group was smeared with the corresponding group of composition (0.1 g / mouse). 20 minutes after xylene induced inflammation, the mice were secondly smeared with deionized water (0.1 ml / mouse) and dexamethasone acetate ointment (0.1 g / mouse) according to the corresponding groups, and the experimental group was smeared with the corresponding group of composition (0.1 g / mouse).

[0054] 30 minutes after the second application of toothpaste, the mice were killed by cervical dislocation, the auricles on both sides were cut off, the test object on the right auricle was cleaned with saline, and the residual saline was absorbed with cotton balls. The auricles on both sides were overlapped, and the left and right ear pieces were punched down at one time with a sharp punch. The ear pieces of both ears were weighed separately, and the swelling value was calculated. The smaller the swelling value, the better the anti-inflammatory effect. The formula is as follows: swelling degree = left ear piece weight - right ear piece weight. The test results are shown in Table 6.

[0055] Table 6 Effects of various compositions containing recombinant human albumin on ear swelling and inflammation in mice

[0056] Note: Compared with the positive control group, P<0.05, * has significant difference.

[0057] It can be seen from Table 6 that compared with the negative control group, the positive control group and experimental groups 1 to 8 have the effect of relieving ear swelling; compared with the positive control group, experimental groups 1 to 2 and experimental groups 7 to 8 can significantly reduce the swelling of the auricle caused by xylene in mice, and have anti-inflammatory effects, among which experimental group 2 has the best effect, which is the best embodiment of the present invention. Experimental groups 5 to 6 have the lowest ability to inhibit ear swelling, further indicating that the use of recombinant human albumin is better than plant-based and animal-based albumin. As can be seen from the results of experiments 5 to 6, the components provided by the present invention are indispensable, and the recombinant human albumin and Lantern Festival extract have obvious synergistic effects, and have good anti-inflammatory, relieving gum redness and pain.

[0058] Example 2 Preparation of recombinant human albumin-containing toothpaste comprising the following components in parts by mass: The effective ingredients are shown in Table 7.

[0059] Table 7 Ingredients of various toothpastes containing recombinant human albumin

[0060] The specific steps for preparing a toothpaste containing recombinant human albumin are as follows: The second aspect of the present invention provides a method for preparing a toothpaste containing recombinant human albumin, comprising the following steps: (1) The Lantern Festival extract is prepared according to the following method: crush the Lantern Festival used in the formula, pass it through a 10-mesh sieve to obtain a powder, soak it in 75% ethanol at a solid-liquid ratio of 1:5 for 6 hours, reflux extract for 15 minutes at a water bath temperature, extract it 3 times, combine the filtrate, distill off the ethanol until there is no alcohol taste, suspend the Lantern Festival extract in deionized water, mix the above solutions evenly, and spray dry the mixed solution to obtain; (2) Inlet air temperature for spray drying: 120~160℃, atomization pressure: 0.1~0.5Mpa, feed rate: 300~700ml / h; (3) Add the Chinese lantern extract, sodium phytate, sodium benzoate, edible flavor, sucralose and sodium saccharin to a certain amount of deionized water according to the mass proportion until they are completely dissolved, which is phase A. Take the xanthan gum, carbomer, sodium carboxymethyl cellulose, sodium lauryl sulfate, sodium lauroyl sarcosinate and mixed powder according to the mass proportion, add them to sorbitol and propylene glycol, place them on a magnetic stirrer and stir at a speed of 400-1000 rpm and a temperature of 50-85°C to make the adhesive uniformly dispersed in the humectant and foaming agent as phase B. Add phase A to phase B. The adhesive rapidly swells into a colloid when it encounters water. Continue stirring until it is mixed and transparent without powdery particles. Store and age for 4 hours, and further stir until the obtained colloid is uniform and transparent, which is phase C. Add silicon dioxide to phase C at a speed of 100-400 rpm to make it fully mixed, and stir at a speed of 100-400 rpm for 30-60 minutes as phase D. After mixing phase D with recombinant human albumin, a high-pressure homogenizer was used for high-pressure homogenization at 300-1000 bar for 30 minutes at a temperature of 25-50°C to obtain a uniform paste; (4) The uniform paste obtained after high-pressure homogenization is placed in a -4°C refrigerator and aged for 24 hours to obtain the recombinant human albumin toothpaste.

[0061] Test Example 6 Antibacterial Test The antibacterial effect of toothpaste was examined with reference to QB / T2738-2012 “Evaluation Methods for Antibacterial and Antimicrobial Effects of Daily Chemical Products” “7.5 Test Methods for Antibacterial Effects of Antibacterial Daily Chemical Products (Inhibitory Ring Method)”.

[0062] The toothpaste stock solution (groups 1 to 9) prepared in Example 2 was prepared with a mass fraction of 30%, separated by centrifugation, and the supernatant was taken and kept at 20°C for 5 min. 50 μl of bacterial suspension (1×10 4 ~ 9×10 4CFU / ml) were placed on the surface of the corresponding nutrient agar medium plate, evenly spread 3 times, covered the plate, and dried at room temperature for 5 minutes. Take sterile dry filter paper, add 20μl of toothpaste stock solution to each piece as the test sample, and take 20μl of sterile distilled water as the control sample (Group 10). Put 3 test pieces on each plate, 1 piece for the photo, cover the plate, seal it with sealing film, place it in a constant temperature incubator at a certain temperature for 16~18h, observe it, use a vernier caliper to accurately measure the diameter of the inhibition zone, and take the average value. When the test bacteria is Staphylococcus aureus, place the culture dish in a constant temperature incubator at 37℃ for 16~18h; when the test bacteria is Candida albicans, place the culture dish in a constant temperature incubator at 28℃ for 16~18h. The test was repeated 3 times. When the test bacteria is Porphyromonas gingivalis, place the culture dish in a constant temperature incubator at 37℃ for 72~96h. The test bacteria is Prevotella intermedia, and the culture dish is placed in a constant temperature incubator at 37℃ for 24~48h. The test bacteria is Forsythia fossili, and the culture dish is placed in a constant temperature incubator at 25℃ for 24~48h. The test bacteria is Fusobacterium nucleatum, and the culture dish is placed in a constant temperature incubator at 37℃ for 72~96h. The test bacteria is Actinobacillus actinomycetemcomitans, and the culture dish is placed in a constant temperature incubator at 37℃ for 24~48h. The test bacteria is Streptococcus mutans, and the culture dish is placed in a constant temperature incubator at 37℃ for 18~24h, and the test bacteria is Lactobacillus, and the culture dish is placed in a constant temperature incubator at 37℃ for 16~24h. The test is repeated 3 times. If the diameter of the antibacterial ring is greater than 7mm, the product has an antibacterial effect, otherwise the product has no antibacterial effect. The control group is sterile deionized water. The antibacterial ring determination effect, the results are shown in Table 8.

[0063] Table 8 Results of inhibition zone determination of antibacterial experiments of various toothpastes containing recombinant human albumin

[0064] Note: “——” No inhibition zone was detected.

[0065] It can be concluded from Table 8 that the recombinant human albumin toothpaste showed good antibacterial activity against Staphylococcus aureus, Candida albicans, Prevotella intermedia, Porphyromonas gingivalis, Steinerella forsythia, Fusobacterium nucleatum, Actinobacillus actinomycetemcomitans and Streptococcus mutans, and the average diameter of the inhibition zone was greater than 7 mm, indicating that the prepared recombinant human albumin toothpaste was moderately sensitive to pathogenic bacteria. The recombinant human albumin toothpaste prepared in Group 4 and Group 3 had the best antibacterial activity; the antibacterial effect gradually increased with the increase of the mass fraction of recombinant human albumin and Lanternaria officinalis (Group 1 and Group 2); but as the mass fraction of recombinant human albumin and Lanternaria officinalis increased to the optimal ratio, the antibacterial effect began to decrease (Group 5 and Group 9); the antibacterial effect of no recombinant human albumin (Group 6) and no Lanternaria officinalis component (Group 7) was much lower than the antibacterial effect of adding both at the same time. It shows that in the recombinant human albumin toothpaste prepared by this method, recombinant human albumin and Lanternaria officinalis are functional ingredients.

[0066] Test Example 7 Anti-caries Test Take the toothpaste prepared by Group 4, Group 3, and Group 2 of Example 2; select 90 subjects, and the inclusion criteria of the subjects are: the subjects are in good general condition; the number of natural teeth in the mouth is ≥15; accompanied by chronic gingivitis; and those who have the habit of brushing teeth in the morning and evening before the test. Exclude those with caries, pulp disease or apical lesions and untreated; those with damaged oral mucosa; those with denture repair, orthodontic appliances, and those with poor restorations in the mouth; those who use antibiotics or tetracycline drugs during the test; those who are allergic to oral cleaning products; those with infectious diseases such as diabetes or hepatitis B. The subjects were randomly divided into 3 groups, 30 in each group, group 1 used the toothpaste prepared by group 4, group 2 used the toothpaste prepared by group 3, and group 3 used the toothpaste prepared by group 2. There was no significant difference in general information such as gender and age among the subjects in each group. The subjects in each group brushed their teeth once in the morning and evening, 2min / time, and used it continuously for 6 weeks. The subjects in each group could not replace other oral cleaning products during the test, and the effect was evaluated after 6 weeks of use. Gum index score: (1) 0 points: healthy gums; (2) 1 point: no inflammation of the gums, but slight changes in gum color and edema, no bleeding after probing; (2) 2 points: slight inflammation of the gums, slight changes in gum color and edema, no bleeding after probing; (3) 3 points: moderate inflammation of the gums, red gums, edema and shiny, bleeding after probing; (4) 4 points: severe inflammation of the gums, obvious redness, swelling and ulceration of the gums, and a tendency to spontaneous bleeding. The test results are shown in Table 9.

[0067] Table 9 The therapeutic effect of the toothpaste of the present invention on gingivitis (score)

[0068] According to the experimental data in Table 9, the toothpaste prepared by the patented technology has a significant improvement effect on the clinical symptoms of patients with severe gingivitis and moderate gingivitis. Studies have shown that the preparation can not only effectively relieve the inflammatory response of gingival tissue, but also achieve tissue repair and functional reconstruction in the gingival lesion area, showing significant anti-inflammatory activity and therapeutic efficacy.

[0069] Test Example 8 Oral ulcer healing experiment Experimental content: A total of 66 clean-grade SD rats (weighing 250-300g) were modeled by intraperitoneal injection of anesthesia phenol cauterization. Criteria for successful modeling: a circular or oval ulcer with a diameter of about 5mm is formed in the cauterized area, surrounded by red and swollen bulges, the mucosal surface is rough and uneven, there is a gray or yellow pseudomembrane, the activity, food intake, and water intake are reduced, and the stool is relatively loose. On the second day after modeling, 66 male SD rats with oral ulcers that were successfully modeled were taken and randomly divided into 11 groups, 6 rats in each group, namely the model group, the drug administration group 1 to the drug administration group 9 (corresponding to the toothpaste prepared in groups 1 to 9 of Example 2, respectively), and the Bingpengsan (commercially available) group, of which the Bingpengsan (commercially available) group was the positive control group. Use a sterile cotton swab to apply the corresponding medicine to the oral ulcer, take the toothpaste prepared in Groups 1 to 9 prepared in Example 2 and the Bingpengsan powder to completely cover the wound surface, apply it once in the morning and evening, with an interval of 8 hours between administrations, and treat continuously for 7 days. The complete healing of the ulcer is regarded as the end point. Observe the changes in the oral mucosa every day, measure the area of ​​the mucosal ulcer without edema, and observe the inflammation and congestion. Detection time points: before application after modeling and 8 hours after the completion of administration. Measure the diameter of the ulcer with a vernier caliper and calculate the ulcer area. At the same time, record the water intake and food intake of each animal per unit time. Evaluation index: record the ulcer area on the 7th day after application: 1mm 2 For healing, ≥1mm 2 The model group is the group that did not heal, and the model group is the group that did not receive any drug. The oral ulcer effect test, the model group, Bingpengsan (commercially available) group, and drug administration group 1 to group 9 had effects on oral ulcers in rats. The results are shown in Table 10.

[0070] Table 10 The therapeutic effect of the toothpaste of the present invention on oral ulcers

[0071] Note: Compared with the model group, P<0.05, * has significant difference.

[0072] According to the experimental data in Table 10, both the drug-administered group and the commercially available group showed an oral wound repair effect compared with the model group, but the efficacy of the single active component (groups 6-7) and the non-optimal ratio preparations (groups 1-2, group 5, combination 8-9) was significantly different, and the wound healing period was extended to more than 5 days. Groups 3-4, which were prepared with the optimal ratio of recombinant human albumin, showed a significant healing effect after 3 days of administration (P<0.05), and their efficacy exceeded that of the traditional Bingpengsan control group. The research data fully confirms the key role of recombinant human albumin combined with Jin Denglong in the treatment of oral ulcers. It can significantly accelerate the repair process of mucosal tissue through the synergistic effect of bioactive ingredients.

[0073] Test Example 9 Antioxidant Experiment-DPPH Scavenging Ability Determination: Preparation of DPPH solution: Accurately weigh 20 mg of DPPH, dissolve it in anhydrous ethanol and make up to 500 ml in a brown volumetric flask to obtain a DPPH solution with a mass fraction of 0.004%. Store it away from light for later use.

[0074] Vitamin C was selected as the positive control. The experimental group (toothpaste prepared in groups 1 to 9 of Example 2) and the positive control solution were prepared with a starting concentration of 1 mg / mL, and the concentration range was 0.002 to 1 mg / mL. The 96-well plate was divided into a sample group and a control group, with 3 replicate wells in each group. A blank control was set for both the sample group and the control group. The blank group was anhydrous ethanol, which was added to the DPPH solution in an equal volume of 1:1. The 96-well plate was reacted in the dark at room temperature for 30 minutes, shaken and evenly mixed, and the absorbance was measured at a wavelength of 517nm. The half inhibitory concentration (IC 50 ) value (μg / ml). IC 50 The IC value is an important indicator to measure the strength of antioxidant activity. It indicates the concentration required for the antioxidant capacity to achieve half of the clearance effect. 50 The smaller the value, the better the free radical scavenging effect. The data results are shown in Table 11, and the calculation formula for free radical scavenging rate is as follows: Free radical scavenging rate (%) = ( )×100% Wherein: A1 is the measured OD value of the sample group, A2 is the measured OD value of the sample control group, and A0 is the measured OD value of the blank group.

[0075] Table 11 DPPH antioxidant treatment results of toothpaste of the present invention

[0076] Note: Compared with vitamin C, P<0.05, * has significant difference.

[0077] According to the experimental data in Table 11, each group (1-9) showed significant free radical scavenging activity, and their DPPH scavenging rates were better than the positive control vitamin C. It is worth noting that the DPPH scavenging efficiency of groups 3-4 was statistically significantly different from that of vitamin C (P<0.05), indicating its strong antioxidant properties. The experiment found that the free radical scavenging rates of group 6 lacking recombinant human albumin, group 7 lacking Lantern Festival extract, and non-optimal ratio groups (groups 1-2, group 5, and group 8-9) were significantly reduced. The results verified that recombinant human albumin and Lantern Festival extract need to be in a specific ratio to achieve an antioxidant synergistic effect.

[0078] Test Example 10 Anti-inflammatory Experiment The effect of xylene on the swelling of the auricle of mice was studied to study its anti-inflammatory effect. Deionized water was used as the negative control group, dexamethasone acetate ointment was used as the positive control group, and toothpastes of groups 1 to 9 prepared in Example 2 were used as test groups 1 to 9. 100 SPF female mice were randomly divided into 11 groups according to their body weight, with 10 mice / group. They were negative control group, positive control group, test group 1, test group 2, test group 3, test group 4, test group 5, test group 6, test group 7, test group 8 and test group 9.

[0079] Xylene was evenly applied to the inner and outer surfaces of the right auricle of each mouse to cause inflammation, with a dose of 100 μl / mouse (50 μl / mouse on one side). The left ear was not treated as a blank control group. 10 minutes after xylene induced inflammation, the mice were first smeared with deionized water (0.1 ml / mouse) and dexamethasone acetate ointment (0.1 g / mouse) according to the corresponding groups, and the experimental group was smeared with toothpaste (0.1 g / mouse) of the corresponding group. 20 minutes after xylene induced inflammation, the mice were secondly smeared with deionized water (0.1 ml / mouse) and dexamethasone acetate ointment (0.1 g / mouse) according to the corresponding groups, and the experimental group was smeared with toothpaste (0.1 g / mouse) of the corresponding group.

[0080] 30 minutes after the second application of toothpaste, the test mice were killed by cervical dislocation, the auricles on both sides were cut off, the test object on the right auricle was cleaned with saline, and the residual saline was absorbed with cotton balls. The bilateral auricles were overlapped, and the left and right ear pieces were punched down at one time with a sharp punch. The ear pieces of both ears were weighed separately, and the swelling value was calculated. The smaller the swelling value, the better the anti-inflammatory effect. The formula is as follows: swelling degree = left ear piece weight - right ear piece weight. The test results are shown in Table 12.

[0081] Table 12 Effect of the toothpaste of the present invention on ear swelling and inflammation in mice

[0082] Note: Compared with the positive control group, P<0.05, * has significant difference.

[0083] According to the experimental data in Table 12, the positive control and test groups (1-9) showed significant inhibitory effects on inflammation compared with the negative control group. It is worth noting that test groups 1-5 had excellent inhibitory effects on xylene-induced ear swelling in mice, which was significantly better than the positive control group, among which test group 4 showed the best anti-inflammatory efficacy. The anti-inflammatory activity of test groups 6-7 was significantly attenuated, and its inhibition rate was the lowest. Recombinant human albumin and Lantern extract can inhibit the activation of inflammatory pathways through synergistic effects, achieving the therapeutic effects of anti-inflammatory, analgesia and tissue repair.

[0084] The above description is only an embodiment of the present invention. The protection scope of the present invention is not limited by these specific embodiments, but is determined by the claims of the present invention. For those skilled in the art, the present invention may have various changes and variations. Any modification, equivalent substitution, improvement, etc. made within the technical ideas and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A composition for oral care containing recombinant human albumin, characterized in that: The composition comprises recombinant human albumin and Lantern Festival Chinese Phalaenopsis extract, wherein the mass fractions of the components are as follows: the mass fraction of recombinant human albumin is 0.5 to 8.0 parts; the mass fraction of Lantern Festival Chinese Phalaenopsis extract is 0.5 to 2.0 parts.

2. The composition according to claim 1, characterized in that The composition further comprises additives acceptable in the oral care product and pharmaceutical fields.

3. The composition according to claim 1, characterized in that The composition further comprises at least one of abrasives, moisturizers, thickeners, foaming agents, anti-sensitivity agents, colorants, flavoring agents, preservatives, whitening agents, fluorides, surfactants, buffers, lubricants, fragrances, pH regulators, and emulsifiers.

4. The use of the composition according to any one of claims 1 to 3, characterized in that: The application includes at least one of the following applications: A1. Application in the preparation of antibacterial products; A2. Application in the preparation of antioxidant products; A3. Application in the preparation of products for inhibiting oral inflammation; A4. Application in the preparation of products for treating and / or preventing dental caries.

5. The use according to claim 4, characterized in that: The product includes at least one of an oral care product and a medicine.

6. The use according to claim 4, characterized in that: The product comprises at least one of toothpaste and mouthwash.

7. A toothpaste containing recombinant human albumin, characterized in that: The toothpaste comprises: the composition according to claim 1; 21.1 to 50.4 parts by mass of a moisturizer; 10 to 25 parts by mass of an abrasive; 1.05 to 7.2 parts by mass of a surfactant; 0.42 to 1.2 parts by mass of a flavoring agent; 0.05 to 0.3 parts by mass of a preservative; 0.02 to 0.20 parts by mass of an emulsifier; and 0.4 to 3.5 parts by mass of a thickener.

8. A method for preparing the composition according to any one of claims 1 to 3, characterized in that: The composition comprises the following steps: S1: Preparation of Lantern Festival extract: crush Lantern Festival, pass through a 10-mesh sieve to obtain powder, soak it in 70%~85% (v / v) ethanol at a material-liquid mass ratio of 1:(3~8), reflux extraction in a water bath at 40~100℃, collect the filtrate, distill the filtrate to remove ethanol until there is no alcohol taste, add deionized water to dissolve and mix the Lantern Festival extract, spray dry the mixed solution to obtain the Lantern Festival extract; S2: Dissolve the Lantern Festival extract in the recombinant human albumin aqueous solution or salt buffer by mass, and use a high-pressure homogenizer at 300-1000 bar for 25-35 minutes at a temperature of 25-50° C. to obtain a uniform solution of the composition.

9. The method according to claim 8, characterized in that The spray drying conditions in step S1 are: air inlet temperature: 120-160° C., atomization pressure: 0.1-0.5 MPa, and feed rate: 300-700 ml / h.

10. The method according to claim 8, characterized in that The method comprises the step of adding additives acceptable in the field of oral care products and medicines to the composition to prepare the oral care products or medicines.

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