Animal oral care composition and its preparation method and application

Abstract

The present invention provides an animal oral care composition and its preparation method and application, relating to the field of animal oral care technology. The animal oral care composition of the present invention is compounded by persimmon extract, licorice chalcone A, and gallnut extract. The three components have a significant synergistic effect and have significant bactericidal and antibacterial effects on animal oral pathogens. It can prevent the formation of dental plaque, prevent inflammatory changes in periodontal tissues, reduce oral odor, remove oral halitosis, freshen breath, and accelerate the healing of oral ulcers. In addition, the animal oral care composition of the present invention can be made into various dosage forms according to different needs, has flexible application methods, uses green and safe raw materials, does not produce resistance and drug resistance after long-term use, is safe and non-toxic to animals, and does not irritate the oral mucosa.

Description

Technical Field

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

[0002] With the development of the economy and the improvement of people's living standards, various pets such as dogs and cats have gradually entered thousands of households, and the health of pets has received more and more attention.

[0003] Due to factors such as pet husbandry practices, oral health issues among pets are gaining increasing attention. The oral cavity of pets (dogs and cats, among others) is composed of hard tissues such as teeth and soft tissues such as the tongue, gums, and periodontal tissues. Their unique dietary habits and lifestyles make them more susceptible to oral diseases, making proper oral hygiene extremely important. According to surveys, over 80% of pets experience oral problems, including bad breath, caries, oral ulcers, dental plaque, tartar, periodontitis, and gingivitis.

[0004] Bad breath, a symptom characterized by an unpleasant odor when breathing or opening the mouth, is quite common in pets (dogs, cats, etc.). Bad breath is divided into oral and non-oral sources. According to statistics, 80% to 90% of bad breath originates from the oral cavity and is caused by oral diseases, such as dental caries, periodontal disease, and oral mucosal diseases. Dental caries and periodontal disease are the most common related diseases. The oral cavity contains a large number of microorganisms that metabolize food residues such as amino acids and proteins remaining in the mouth, producing unpleasant odors - volatile sulfur compounds (VSCs). The main components of these compounds are hydrogen sulfide, methyl mercaptan, and ethyl sulfide. Studies have shown that inhibiting the proliferation of Porphyromonas gingivalis, Prevotella intermedia, and Fusobacterium nucleatum can reduce the level of volatile sulfides and reduce bad breath in animals.

[0005] Dental caries, commonly known as tooth decay or cavities, is a bacterial disease that can cause pulpitis and periapical periodontitis, and even inflammation of the alveolar bone and jawbone. If dental caries are not treated promptly, the lesions will continue to progress, forming cavities that eventually destroy and disappear the tooth crown, ultimately leading to tooth loss. Bacteria are essential for the development of dental caries, and the primary pathogenic bacteria in dental caries is currently believed to be Streptococcus mutans, which forms a dental plaque biofilm on the tooth surface, providing a microecological environment for the accumulation and growth of pathogenic bacteria. Therefore, inhibiting and eliminating Streptococcus mutans is a crucial step in the prevention and treatment of dental caries.

[0006] Periodontal disease refers to diseases of the tooth-supporting tissues (periodontal tissues). It can be categorized as gingivitis, which affects only the gum tissue, and periodontitis, which affects the deeper periodontal tissues (periodontal ligament, alveolar bone, and cementum). Periodontal disease is a common oral disease in pets (dogs, cats, and others). When clean teeth are exposed to saliva, a film of glycoproteins immediately forms. Oral bacteria adhere to this film and begin to multiply, forming dental plaque. Food particles, sloughed epithelial cells, and saliva can also contribute to the formation of dental plaque. Dental plaque forms very quickly in animals, typically several hours after teeth cleaning. Plaque is a soft substance that cannot be removed by tongue movement, drinking water, or saliva. Plaque that is not removed promptly eventually transforms into tartar, which accumulates above or near the gum line. Since the oral environment of dogs and cats is generally alkaline (compared to the acidic environment of humans), tartar can develop into calculus within just 3 to 5 days. Therefore, the incidence of tartar in pets is over five times that of humans. At present, dental plaque is recognized as the initiating factor of periodontal disease and the main pathogenic factor causing periodontal disease. The imbalance of the bacterial flora in dental plaque leads to the disruption of the balance between the bacterial flora and the host immune defense, which in turn causes inflammatory changes in the periodontal tissues. Gingivitis is a precursor to periodontitis and is caused by bacterial biofilm (dental plaque) that accumulates on the teeth near the gums (gingiva). Gingivitis is a reversible, non-destructive form of periodontitis that causes inflammation of the supporting tooth tissue but does not cause shedding or bone loss. Periodontitis occurs when the inflammatory process leads to loss of attachment to the periodontal ligament (PDL) of the dentition, gingival atrophy and ultimately bone loss. Studies have shown that the main pathogens causing periodontal disease are Porphyromonas gingivalis and Fusobacterium nucleatum.

[0007] Ulcerative stomatitis is a common clinical disease in dogs and cats, characterized by redness, swelling, ulcers, pain on the tongue, gums, buccal mucosa, and lips, inability to chew, and excessive salivation.

[0008] The oral problems mentioned above in pets (dogs, cats, etc.) are primarily caused by pathogenic bacteria. Therefore, the ultimate goal of preventing and treating oral problems is to inhibit the growth of oral bacteria by controlling Streptococcus mutans, Porphyromonas gingivalis, Fusobacterium nucleatum, and Prevotella intermedia.

[0009] However, most pet oral care products currently claiming antibacterial and anti-inflammatory properties use chemical antiseptics, such as chlorhexidine and cetylpyridinium chloride, as their active ingredients. This poses safety concerns and is prone to developing resistance and drug tolerance, hindering the effectiveness of animal oral care and posing a health risk. Therefore, providing an oral care product made from natural plant materials and simple components that can alleviate problems such as tooth decay, bad breath, dental plaque, oral ulcers, and periodontal disease in pets, without developing resistance and drug tolerance with long-term use, while also being safe, non-toxic, and non-irritating to the oral mucosa, remains an urgent technical challenge in this field. Summary of the Invention

[0010] The animal oral care composition of the present invention comprises the following raw materials in parts by weight: 1 to 6 parts of persimmon extract, 0.3 to 1 part of licorice chalcone A and 3 to 6 parts of gallnut extract.

[0011] Furthermore, the animal oral care composition comprises the following raw materials in parts by weight: 5 parts of persimmon extract, 0.5 parts of licorice chalcone A and 5 parts of gallnut extract.

[0012] Furthermore, the persimmon extract is extracted from the fruit of the persimmon tree.

[0013] Furthermore, the persimmon tannin content in the persimmon extract is ≥75 wt%.

[0014] Furthermore, the persimmon extract is food grade persimmon extract.

[0015] Persimmon extract is obtained from the fruit of the persimmon tree, Diospyros kaki Thunberg (Ebenaceae). Persimmon tannin is the main component of persimmon extract. By adding persimmon extract to the product, the present invention eliminates bad breath-causing bacteria (Porphyromonas gingivalis) and tooth decay-causing bacteria (Streptococcus mutans), improving oral health and freshening breath in animals. It also has an excellent deodorizing effect on sulfides and methyl mercaptan, which are responsible for bad breath.

[0016] Furthermore, the licorice chalcone A is extracted from the root of Glycyrrhiza inflata.

[0017] Furthermore, the purity of the licorice chalcone A is ≥95%.

[0018] Furthermore, the licorice chalcone A is food-grade licorice chalcone A or pharmaceutical-grade licorice chalcone A.

[0019] Licorice (Glycyrrhiza L.) is a traditional Chinese medicinal material with a history of more than 2,000 years of medicinal use in China. There are many types of licorice, the main ones being: Ural licorice (Glycyrrhiza uralensis Fisch.), licorice glabra (Glycyrrhiza glabra L.), and licorice inflata Bat. Licorice contains a variety of bioactive ingredients, including glycyrrhizic acid, glycyrrhetinic acid, glycyrrhizin, isoglycyrrhizin, glycyrrhizin flavonoids, and polysaccharides. At present, more than 150 glycyrrhizin flavonoids have been discovered by scientific research, including licorice chalcone A (molecular formula: C 21 H 22O4, molecular weight: 338.4) is a flavonoid compound with multiple physiological activities. The present invention adds licorice chalcone A to the product to make the animal oral care composition have good antibacterial, anti-ulcer and anti-inflammatory effects.

[0020] Furthermore, the gallnut extract is obtained by supercritical CO2 extraction.

[0021] Furthermore, the gallnut extract is a food-grade gallnut extract or a pharmaceutical-grade gallnut extract.

[0022] Gallnut extract is obtained by extracting cystic galls formed by female aphids (Aphididae) parasitizing the tender leaves or petioles of the Anacardiaceae plant "Rhus chinensis" and other plants of the same genus. Gallnut contains tannins (tannic acid), gallic acid, methyl gallate, and ethyl gallate. The present invention uses gallnut extract to impart antibacterial, anti-caries, anti-inflammatory, and analgesic effects to an animal oral care composition, and has a certain therapeutic effect on oral ulcers. Furthermore, the gallnut extract of the present invention also has an inhibitory effect on oral cariogenic bacteria.

[0023] The preparation method of the animal oral care composition of the present invention is as follows: persimmon extract, licorice chalcone A and gallnut extract are uniformly mixed to obtain the composition.

[0024] Another object of the present invention is to provide a use of the animal oral care composition in a product for maintaining animal oral health.

[0025] Furthermore, the product is one or more of oral spray, mouthwash, tooth cleaning powder, tooth gel, and toothpaste.

[0026] Compared with the prior art, the present invention has the following beneficial technical effects:

[0027] 1. This invention is the first to scientifically combine persimmon extract, licorice chalcone A, and gallnut extract. The three ingredients have a significant synergistic effect, demonstrating excellent antibacterial and deodorizing effects in animal oral care. Furthermore, when the three ingredients are used together, the antibacterial and deodorizing effects of the product are significantly enhanced compared to when used alone.

[0028] 2. The animal oral care composition of the present invention has significant bactericidal and antibacterial effects on oral pathogens, such as Streptococcus mutans, Bacillus nucleatum, Porphyromonas gingivalis, and Prevotella intermedia. It can inhibit oral pathogens from adhering to and multiplying on glycoprotein films, thereby preventing the formation of dental plaque.

[0029] 3. The oral care composition for animals of the present invention can effectively prevent inflammatory changes in periodontal tissues caused by dysbiosis in dental plaque and disruption of the balance between the flora and host immune defense;

[0030] 4. The oral care composition for animals of the present invention can effectively remove volatile sulfides such as hydrogen sulfide and methyl mercaptan from the oral cavity of animals, inhibit the proliferation of Porphyromonas gingivalis, Prevotella intermedia, and Fusobacterium nucleatum, reduce oral odor, eliminate oral halitosis, and freshen breath;

[0031] 5. The oral care composition for animals of the present invention is beneficial to epithelial cell regeneration and accelerates the healing of oral ulcers;

[0032] 6. The oral care composition for animals of the present invention can be prepared into various dosage forms according to different needs, has flexible application methods, uses green and safe raw materials, does not add chemical sterilization ingredients, does not produce resistance and drug resistance after long-term use, is safe and non-toxic to animals, has no irritation to the oral mucosa, and has a relieving effect on animal oral diseases, such as oral caries, bad breath, oral ulcers, and periodontal tissue lesions. DETAILED DESCRIPTION

[0033] The technical solution provided by the present invention is further described below in conjunction with embodiments.

[0034] Example 1

[0035] An animal oral care composition, comprising the following raw materials in parts by weight:

[0036] 5 parts of persimmon extract, 0.5 parts of licorice chalcone A, and 5 parts of gallnut extract.

[0037] The preparation method of the animal oral care composition is as follows: uniformly mix persimmon extract, licorice chalcone A and gallnut extract to obtain the composition.

[0038] Example 2

[0039] An animal oral care composition, comprising the following raw materials in parts by weight:

[0040] 6 parts of persimmon extract, 1 part of licorice chalcone A, and 6 parts of gallnut extract.

[0041] The preparation method of the animal oral care composition is as follows: uniformly mix persimmon extract, licorice chalcone A and gallnut extract to obtain the composition.

[0042] Example 3

[0043] Provided is an oral care composition for animals, the composition and proportion (in parts by weight) are as follows:

[0044] 1 part of persimmon extract, 0.3 parts of licorice chalcone A, and 3 parts of gallnut extract.

[0045] The preparation method of the animal oral care composition is as follows: uniformly mix persimmon extract, licorice chalcone A and gallnut extract to obtain the composition.

[0046] Comparative Example 1

[0047] Same as Example 1, except that the composition and proportion do not include persimmon extract, and the reduced portion is proportionally allocated to licochalcone A and gallnut extract.

[0048] Comparative Example 2

[0049] Same as Example 1, except that the composition and proportion do not contain licochalcone A, and the reduced portion is proportionally distributed to persimmon extract and gallnut extract.

[0050] Comparative Example 3

[0051] Same as Example 1, except that the composition and proportion do not include the gallnut extract, and the reduced portion is proportionally allocated to the persimmon extract and licorice chalcone A.

[0052] Comparative Example 4

[0053] 10.5 parts of Gallnut Extract.

[0054] Comparative Example 5

[0055] 10.5 parts of Licochalcone A.

[0056] Comparative Example 6

[0057] 10.5 parts persimmon extract.

[0058] Test Example 1 In vitro antibacterial test

[0059] Minimum inhibitory concentration (MIC) test

[0060] 1. Principle

[0061] In this test, different concentrations of antibacterial agents are mixed and dissolved in nutrient broth culture medium, and then bacteria are inoculated. The minimum inhibitory concentration (MIC) of the antibacterial agent that inhibits the growth of the test bacteria is determined by whether the bacteria grow or not.

[0062] 2. Test materials

[0063] Strains: Porphyromonas gingivalis (ATCC33277); Streptococcus mutans (ATCC25175); Fusobacterium nucleatum (ATCC49256); Actinomyces viscosus (ATCC15987); Prevotella intermedia (ATCC25611).

[0064] Samples to be tested: oral care compositions prepared in Examples 1-3 and oral care compositions prepared in Comparative Examples 1-6.

[0065] 3. Reagents, equipment and operating procedures

[0066] Refer to the minimum inhibitory concentration test (nutrient broth dilution method) in the "Disinfection Technical Specifications (2002 Edition)".

[0067] 4. Judging Rules

[0068] When the positive control tube has bacterial growth (turbidity) and the negative control tube has no bacterial growth (transparent), the effective concentration of the test bacterial suspension is 5×10 5 cfu / mL~5×10 6 cfu / mL, the antibacterial (inhibitory) agent concentration corresponding to the highest dilution of sterile growth in the test group is the MIC of the sample to the test bacteria.

[0069] 5. Test results

[0070]

[0071] Conclusion: As can be seen from the above table, the oral care compositions for animals prepared in Examples 1-3 and Comparative Examples 1-6 all have antibacterial effects, but the antibacterial effects of Examples 1-3 are significantly better than those of Comparative Examples 1-6, indicating that the compounding of the raw materials of the present invention can play a synergistic role and significantly enhance the antibacterial effect of the product.

[0072] At the same time, the antibacterial effects of Example 1 and Example 2 are similar, but the amount of licorice chalcone A in Example 1 is reduced by half. It can be seen that the reasonable compounding of products can not only improve the antibacterial effect, but also save the use of raw materials and reduce production costs.

[0073] Test Example 2 Bad breath improvement performance test 1

[0074] 1. Test materials

[0075] Source of bacteria: A complete saliva bacterial specimen was collected from a dog with periodontal disease and halitosis in an animal hospital. The VSCs value was 205 ppb (normal value is 125 ppb). The saliva was subcultured in liquid anaerobic culture medium and then preserved.

[0076] Samples to be tested: The oral care compositions prepared in Examples 1-3 and the oral care compositions prepared in Comparative Examples 1-6 were respectively prepared with sterile water to prepare solutions (5 g / 100 mL).

[0077] 2. Reagents and equipment

[0078] Test reagents: anaerobic liquid culture medium.

[0079] Test instruments: Halimeter VSCs measuring instrument (RH-17 model), anaerobic glove box.

[0080] 3. Test methods

[0081] (1) Bacterial culture: Saliva samples from clinical dogs with periodontal disease and halitosis were activated and cultured in anaerobic liquid culture medium (15% N2, 75% CO2, 10% H2) at 37°C for 48 hours to a concentration of 105-106 CFU / mL. 100 μL of the above bacterial solution and 100 μL of the test sample were added to a test tube containing 2 mL of anaerobic liquid culture medium and cultured anaerobically for 48 hours. A positive control was established in which only the bacterial solution and an equal amount of liquid culture medium were added without the test sample.

[0082] (2) VSCs value determination: Determine using the Halimeter VSCs meter. There are two latex tubes of different lengths on the silicone stopper. Connect one end of the test tube (the longer one) to the latex tube of the Halimeter VSCs meter, and insert the other end into the test tube about 2 mm from the surface of the bacterial solution. The other latex tube is the air inlet tube. Before testing the VSCs value, insert the silicone stopper into the bacterial solution containing the sample to be tested, and shake the test tube twice while measuring each VSCs value. Do this a total of 6 times, remove the base value, take the peak value, and finally calculate the average value.

[0083] (3) Statistical analysis: SPSS 10.0 software was used to perform correlation analysis on VSCs values.

[0084] 4. Test results

[0085] The VSCs value of the positive control was (180 ± 82) ppb. As shown in the table, the animal oral care composition aqueous solutions (5 g / 100 mL) of Examples 1-3 significantly reduced VSCs, outperforming the animal oral care composition aqueous solutions (5 g / 100 mL) of Comparative Examples 1-6, demonstrating that the combination of the ingredients can synergize and significantly enhance the deodorizing effect. Furthermore, Example 1 exhibited a very strong deodorizing effect, achieving the same deodorizing effect as Example 2.

[0086] Samples to be tested VSCs value (ppb) Example 1 24±11.05 Example 2 23.75±9.75 Example 3 38.5±15.9 Comparative Example 1 119.0±45.36 Comparative Example 2 74.95±23.76 Comparative Example 3 118.0±46.12 Comparative Example 4 75.25±21.36 Comparative Example 5 151.0±64.31 Comparative Example 6 74.25±24.01

[0087] Bad breath improvement performance test 2

[0088] 1. Materials and Animals

[0089] Test samples: the oral care compositions prepared in Examples 1-3 were respectively mixed with sterile water to form solutions (3 g / 100 mL); chlorhexidine gluconate (0.12 wt %); and deionized water (control group) were used as test samples.

[0090] Experimental animals: 60 dogs of the same species, age, and breeding conditions, half male and half female

[0091] 2. Test methods

[0092] 60 pet dogs with symptoms such as bad breath and gingivitis were randomly divided into 5 groups.

[0093] Each group consisted of 12 rats, and their oral conditions were scored. The oral care compositions prepared in Examples 1-3 of the test samples were mixed with sterile water to form a solution (3 g / 100 mL); chlorhexidine gluconate (0.12 wt %); and deionized water (control group) were placed in a prepared spray bottle. Each group then used the solution for two consecutive weeks, twice a day, 2 mL each time. After two weeks, the test was scored again, and the scores and their averages were calculated.

[0094] 3. Test results

[0095] The Rosenberg scoring system was used, with lower scores indicating less severe bad breath. The scores were evaluated before and after the test using the following criteria:

[0096] Score describe 0 No bad breath, definitely no odor 1 Suspicious bad breath, with a strange smell and odor 2 Slight bad breath, definitely smells bad but very light 3 Moderate bad breath, obvious bad breath 4 Severe bad breath, but still tolerable to the examiner 5 Strong odor that the examiner cannot tolerate

[0097] The test results are as follows:

[0098]

[0099]

[0100] As can be seen from the table, test groups 1-4 had significant effects on removing and improving oral odor. Test groups 1 and 2 had better oral odor removal effects than test group 4. Test group 5 (control group) with deionized water had no significant effect.

[0101] Conclusion: The oral care compositions for animals prepared in Examples 1-3 of the present invention significantly inhibit and improve oral halitosis, demonstrating that the combination of the raw materials can synergize and significantly enhance the deodorizing effect. Furthermore, the deodorizing effects of Examples 1 and 2 were very strong, and the deodorizing effects of the oral care compositions for animals prepared in Examples 1 and 2 (3 g / 100 mL) were superior to those of chlorhexidine gluconate (0.12 wt%).

[0102] Test Example 3: Dental Caries Prevention and Treatment Test

[0103] 1. Experimental materials and animals:

[0104] Test samples: oral care compositions prepared in Examples 1-3 and oral care compositions prepared in Comparative Examples 1-6 were respectively prepared with sterile water to form solutions (3 g / 100 mL); chlorhexidine gluconate (0.12 wt %); and deionized water (negative control group) were used as the samples to be tested.

[0105] Test strain: Streptococcus mutans (ATCC25175) was provided by Shanghai Collection Biotechnology Center.

[0106] Experimental animals: 110 SPF-SD rats, half male and half female, all weaned 17 days after birth, with good mobility, no dental caries, and no periodontal disease.

[0107] Breeding unit: Jinan Pengyue Experimental Animal Breeding Co., Ltd.

[0108] 2. Establishment of rat caries model

[0109] One hundred and ten SPF-SD rats were weaned 17 days after birth. Body weight was recorded and the rats were fed a cariogenic diet consisting of Keyes 2000# and 5% sucrose solution. Ampicillin sodium was administered from 18 to 20 days of age for bacteriostasis. Saliva was collected with a sterile cotton swab and plated on a microsalivary-bacitracin agar (MSB) plate to assess the inhibition of endogenous mutans Streptococcus. From 22 to 25 days of age, ATCC 25175 (1 mL / rat) was inoculated intraorally for four consecutive days. Successful oral colonization with mutans Streptococcus was confirmed by testing on days 23 to 25 for subsequent experiments.

[0110] 3. Animal grouping test

[0111] 110 rats were randomly divided into 11 groups (10 rats per group). Starting on day 28, the teeth of rats in groups 1-11 were treated with a cotton swab containing a sterile aqueous solution (3 g / 100 mL) of the oral care composition prepared in Examples 1-3 and Comparative Examples 1-6, 0.12 wt% chlorhexidine gluconate, and deionized water (negative control group). After treatment, rats were deprived of food for 2 hours. Treatment was repeated twice daily for 5 consecutive weeks. Body weight of the rats was recorded weekly, and their health status was recorded daily.

[0112] On day 61, 100 μL of stimulated saliva was collected from each rat and smeared on bovine heart and brain infusion (BHI) blood agar (total colony count) and MSB agar (total S. mutans colony count). The cells were incubated anaerobically at 37°C for 48 hours, and the number of S. mutans colonies and total colony counts were recorded. The level of S. mutans in rat saliva was expressed as the ratio of the number of MSB colonies to the number of BHI colonies [S. mutans level (%) = number of MSB colonies / number of BHI colonies × 100%].

[0113] On day 63, the rats were sacrificed, and jawbone specimens were removed, cleaned, dried, and stained. Ultrathin diamond shavings were used to sagittally section the maxillary and mandibular molar surfaces along the mesiodistal plane. The smooth surface and pit and fissure caries lesions of the molars were graded and scored using the Keyes scoring method under a stereomicroscope to assess caries inhibition in the experimental group. This process was performed blindly by two other experimenters (one recording the Keyes score and the other verifying it).

[0114] Caries inhibition rate (%) = (caries score of negative control group - caries score of experimental group) / caries score of negative control group × 100%.

[0115] During the experiment, no significant differences in food and water consumption were observed between the groups, ensuring that all groups were exposed to the same cariogenic attack. During the treatment period, the rats showed no abnormalities such as erythema or swelling in the oral mucosa. At the beginning of the experiment, there was no statistically significant difference in body weight between the groups; during the experiment, the rats gained weight uniformly; and at the end of the experiment, there was no statistically significant difference in body weight gain between the groups.

[0116] 4. Test results

[0117] Group Test samples n Streptococcus mutans level (%) Experimental group 1 Example 1 (3 g / 100 mL) 12 20.6.±2.6 Experimental Group 2 Example 2 (3 g / 100 mL) 12 20.3±2.3 Experimental Group 3 Example 3 (3g / 100mL) 12 22.8±2.5 Experimental Group 4 Comparative Example 1 (3 g / 100 mL) 12 27.2±2.8 Experimental Group 5 Comparative Example 2 (3 g / 100 mL) 12 25.3±2.2 Experimental Group 6 Comparative Example 3 (3 g / 100 mL) 12 26.5±2.7 Experimental Group 7 Comparative Example 4 (3 g / 100 mL) 12 25.6±2.3 Experimental Group 8 Comparative Example 5 (3 g / 100 mL) 12 29.6±2.9 Experimental Group 9 Comparative Example 6 (3 g / 100 mL) 12 25.0±2.3 Experimental group 10 0.12wt% chlorhexidine gluconate 12 20.5±2.2 Experimental group 11 Deionized water (negative control group) 12 32.1±3.2

[0118] As can be seen from the table, at the end of the experiment (61st day), the levels of Streptococcus mutans in experimental groups 1-3 and experimental group 10 were reduced to varying degrees, among which the levels of experimental groups 1, 2 and 10 were close.

[0119] Rat dental caries scoring

[0120] Keyes scoring for smooth surface caries and pit and fissure caries: According to Keyes' classic method for evaluating dental caries, the degree of dental caries damage is divided into four grades: grade E caries, caries only accumulates on the enamel; grade Ds caries, enamel caries and caries lesions do not exceed 1 / 4 of the outer layer of dentin; grade Dm caries, caries lesions involve 1 / 4 to 3 / 4 of the dentin thickness; grade Dx caries, caries lesions involve more than 3 / 4 of the dentin thickness.

[0121] Dental caries scores of rats in each group

[0122]

[0123] Conclusion: The oral care compositions prepared in Examples 1-3 exhibited significantly better caries-inhibiting effects than the oral care compositions prepared in Comparative Examples 1-6, demonstrating that the combination of the ingredients can synergistically enhance caries inhibition. Furthermore, the caries-inhibiting effects of Experimental Groups 1, 2, and 10 were very similar.

[0124] Test Example 4: Gingivitis Reduction Test

[0125] 1. Materials and Animals

[0126] Test samples: oral care compositions prepared in Examples 1-3 and Comparative Examples 1-6 were respectively mixed with sterile water to form solutions (3 g / 100 mL); chlorhexidine gluconate (0.12 wt %); and deionized water (control group) were used as test samples.

[0127] Experimental animals: 110 dogs, half male and half female

[0128] 2. Test methods

[0129] 110 pet dogs suffering from symptoms such as bad breath and gingivitis were selected and randomly divided into 11 groups of 10 dogs each. Their oral conditions were scored. The oral care compositions prepared from the test samples Examples 1-3 and Comparative Examples 1-6 were respectively prepared with sterile water to form a solution (3 g / 100 mL); chlorhexidine gluconate (0.12 wt %); and deionized water (control group) were placed in a prepared spray bottle. Each group then used the solution for two consecutive weeks, twice a day, 2 mL each time. After two weeks, the test was scored again, and the scores and their averages were calculated.

[0130] 3. Test results

[0131] Gingival Index (GI): Check the condition of each dog's gums and record the tooth surface score and its average value. The lower the score, the healthier the gums.

[0132] Scoring Criteria

[0133] Score describe 0 Gum health 1 Mild gingival inflammation, slight change in gum color and slight edema, no bleeding on probing 2 Moderate gingival inflammation, red gums, edema, shiny gums, bleeding on probing 3 Severe gingivitis, with noticeable redness and swelling of the gums, accompanied by ulcers and a tendency to spontaneous bleeding

[0134] Test results

[0135]

[0136]

[0137] As can be seen from the table, test groups 1-3 and test group 10 have a significant effect on alleviating oral gingivitis, which is better than test groups 4-9.

[0138] Conclusion: The oral care compositions for animals prepared in Examples 1-3 are significantly better than the oral care compositions for animals prepared in Comparative Examples 1-6 in alleviating oral gum symptoms, indicating that the compounding of the raw materials can exert a synergistic effect and significantly enhance the therapeutic effect of gum symptoms.

[0139] Test Example 5: Dental Plaque Inhibition Test

[0140] 1. Materials and Animals

[0141] Test samples: The oral care compositions prepared in Examples 1-3 were respectively mixed with sterile water to form solutions (3 g / 100 mL); chlorhexidine gluconate (0.12 wt %) was used as the test sample.

[0142] Experimental animals: 10 dogs were raised, half of them were male and half were female.

[0143] 2. Test methods

[0144] Ten pet dogs were randomly divided into five groups of two dogs each, with a control group included. The test dogs in the test sample-treated group underwent unilateral ultrasonic cleaning to observe the effects of the oral care compositions (3 g / 100 mL) and chlorhexidine gluconate (0.12 wt %) prepared in Examples 1-3 on clean teeth and teeth with plaque. The test dogs in the control group also underwent unilateral ultrasonic cleaning to observe the growth of dental plaque on clean and plaque-bearing teeth without the test sample treatment.

[0145] All five groups of experimental dogs were anesthetized and underwent unilateral ultrasonic dental cleaning on the left buccal teeth of the maxillary and mandibular teeth of the experimental dogs. The teeth were cleaned until the dental plaque index was determined to be 0. The teeth of the experimental dogs were then stained with a plaque developer, and the unilateral dental plaque index of the experimental dogs was determined using the Quigley-Hein method. The average dental plaque index of the maxillary and mandibular teeth was calculated.

[0146] Plaque index determination

[0147]

[0148]

[0149] Ten experimental dogs were housed in the same environment, fed the same brand and amount of dog food and water. Four hours after each meal, the teeth of the experimental sample-treated dogs were sprayed with 2 ml of the oral care composition prepared in Examples 1-3 (3 g / 100 mL) and chlorhexidine gluconate (0.12 wt %). The control group received no treatment. Four weeks later, the five experimental dogs were again anesthetized with respiratory anesthetics. Plaque staining was performed on the teeth of the experimental dogs on one side using the Quigley-Hein method, and the results were compared to those four weeks prior.

[0150] This experiment used SPSS20.0 statistical software to perform statistical analysis on the collected data.

[0151] 3. Test results

[0152]

[0153] As shown in the table, the amount of dental plaque on the tooth surfaces of the dogs in test groups 1-4, which were cleaned, was significantly less than that of the dogs in the control group. The amount of dental plaque on the tooth surfaces of the dogs in test groups 1-4, which were cleaned, was not significantly different from that of the dogs in the control group. Test groups 1-4 showed a significant effect in inhibiting dental plaque on clean teeth, effectively alleviating plaque accumulation. Furthermore, test groups 1, 2, and 4 demonstrated comparable plaque inhibition.

[0154] Conclusion: The oral care compositions prepared in Examples 1-3 have a significant inhibitory effect on the formation of dental plaque and can effectively alleviate the accumulation of dental plaque on teeth.

[0155] Comparative Example 6 Oral Ulcer Treatment Test

[0156] 1. Materials and Animals

[0157] Test sample: Example 1-3 Preparation of oral care composition for animals

[0158] Experimental animals: 36 SPF-SD rats, half male and half female, weighing 200-250 g, well-active, free of dental caries and periodontal disease. Breeding unit: Jinan Pengyue Experimental Animal Breeding Co., Ltd.

[0159] 2. Establishment of oral ulcer animal model

[0160] Thirty SPF-SD rats were anesthetized with 3 mL / kg of 10% chloral hydrate solution via intraperitoneal injection. After anesthesia, a 6-cm-long double-ended glass tube with an inner diameter of 60 mm was placed inside the tube. A small cotton ball was placed at one end, with the bottom of the cotton ball flush with the tube opening. 40% glacial acetic acid solution was dripped into the tube to saturate the cotton ball. The open end of the glass tube with the cotton ball was then placed flat against the right buccal mucosa of the rat. After 90 seconds of cauterization, large, irregular white lesions appeared in this area. 24 hours later, a circular or oval ulcer developed in the experimental area on the right cheek of the rat, thus establishing a rat oral ulcer model.

[0161] 3. Test methods

[0162] On the second day, 30 rats from the successfully established models were randomly divided into a blank group (0.9 wt% saline), a test group (2.0 g / kg of the composition), a control group (1.0 g / kg of Guilin Watermelon Frost), and a healthy group (0.9 wt% saline). The drug was applied to the oral ulcers of the rats four times daily, 3 hours apart, for 10 consecutive days. The maximum transverse diameter (d1) and maximum longitudinal diameter (d2) of the oral ulcers in each group of rats were measured with a vernier caliper before dosing and on days 4, 6, and 10 after dosing. The ulcer area was calculated (ulcer area = π × d1 × d2 × 1 / 4, where π is 3.14).

[0163] Oral ulcer area of ​​rats

[0164]

[0165]

[0166] As shown in the table, compared with the blank group, the oral ulcer areas of rats in Examples 1-3 and Guilin Watermelon Frost groups were significantly reduced, indicating that the oral care compositions for animals prepared in Examples 1-3 have significant therapeutic effects on oral ulcers.

[0167] Conclusion: The oral care compositions for animals prepared in Examples 1-3 have significant therapeutic effects on oral ulcers. The therapeutic effects of Example 1 and Example 2 are comparable.

[0168] Test Example 7 Oral mucosal irritation test

[0169] 1. Materials and Animals

[0170] Test sample: Example 1 Preparation of oral care composition for animals

[0171] Experimental animals: 3 SPF male golden hamsters, 60-65 days old, bred by Beijing Weitonglihua Experimental Animal Technology Co., Ltd.

[0172] 2. Test methods

[0173] The test was conducted in accordance with the method in "YY / T 0127.13-2018 Biological Evaluation of Oral Medical Devices Part 13: Oral Mucosal Irritation Test." Three male golden hamsters were sampled. A 10mm diameter cotton ball was soaked with the test sample and placed in the hamster's left cheek pouch. A cotton ball soaked in physiological saline was placed in the other cheek pouch as a control. The contact time was 5 minutes, repeated once every hour for a total of 4 times. The cheek pouch was inspected after each contact and before repeated placement. The cheek pouch was observed visually 24 hours after the last contact. At the end of the test, the animals were sacrificed using an overdose of anesthesia. The mucosa and surrounding tissues at the sample contact site were collected, fixed with 10% formalin, embedded in conventional paraffin, and semi-serial sectioned, with 5 sections taken at intervals, and stained with hematoxylin and eosin.

[0174] 3. Test evaluation

[0175] Visual Observation and Scoring: After euthanasia, the cheek pouch mucosa was visually inspected for irritation, injury, congestion, swelling, erosion, or ulceration. The cheek pouches on the experimental and control sides of the same animal were compared, and the condition of each cheek pouch was recorded, noting any differences between the experimental and control sides. Oral mucosal clinical manifestations were scored according to the oral mucosal reaction scoring system in Table 1, 10.1.2, of "YY / T 0127.13-2018 Biological Evaluation of Oral Medical Devices - Part 13: Oral Mucosal Irritation Test." The average score for each animal was calculated by summing the scores of each animal in each observation period and dividing by the total number of animals observed.

[0176] Histological Evaluation and Scoring: Microscopically evaluate the irritation response of mucosal tissue. Observe the epithelium and connective tissue for signs of hyperkeratosis, dyskeratosis, acanthosis, laxity, epithelial dysplasia, epithelial atrophy, spongiosis, basal cell vacuolation, liquefaction, metaplasia, hyperplasia, and other abnormal changes. Calculate the tissue response score for each animal according to Table 2, Section 10.2.2 of "YY / T0127.13-2018 Biological Evaluation of Oral Medical Devices - Part 13: Oral Mucosal Irritation Test" for the oral mucosal tissue response scoring system. Add the microscopic evaluation scores of all animals in the experimental group and divide by the total number of observations to calculate the average score for the experimental group. The same calculation was performed for the control group. The maximum score for each group was 16.

[0177] 4. Test results

[0178] Results of golden hamster oral mucosal reaction scoring:

[0179]

[0180] The stimulation index was obtained by subtracting the mean score of the control group from the mean score of the experimental group.

[0181] Results of golden hamster oral mucosal tissue reaction scoring:

[0182]

[0183] The degree of oral mucosal tissue reaction was graded according to the irritation index.

[0184] Oral mucosal tissue reaction grading

[0185]

[0186]

[0187] Oral mucosal irritation index: 0, reaction degree: none.

[0188] Conclusion: The results of the irritation test on the oral mucosa of golden hamsters of the oral care composition for animals prepared in Example 1 were: non-irritating.

[0189] Test Example 8 Toxicology Test

[0190] 1. Materials and Animals

[0191] Test sample: Example 1 Preparation of oral care composition for animals

[0192] Experimental animals: mice

[0193] Level: SPF level

[0194] Number and gender: 20, half male and half female

[0195] Weight: 18~22g

[0196] Breeding unit: Jinan Pengyue Experimental Animal Breeding Co., Ltd.

[0197] 2. Test methods

[0198] Refer to 2.3.1 Acute oral toxicity test of "Technical Specifications for Disinfection" (2002 edition).

[0199] 3. Test results

[0200] The KM mice showed no abnormal symptoms or death within 14 days of exposure. After the end of the experimental observation, the animals were subjected to a gross autopsy and no abnormalities were found. The acute oral toxicity LD50 of this sample to KM mice was >5000 mg / kg·bw. The results are as follows:

[0201] Acute oral toxicity test results

[0202]

[0203] Conclusion: The oral care composition for animals prepared in Example 1 has an acute oral toxicity LD50 of >5000 mg / kg·bw to KM mice, and is practically non-toxic in the acute oral toxicity test.

[0204] In summary, the oral care composition for animals of the present invention can effectively prevent and treat gingivitis and oral ulcers, prevent dental plaque and dental caries, relieve bad breath, and freshen breath.

[0205] The animal oral care composition of the present invention can be prepared into various dosage forms such as pet mouthwash, toothpaste, tooth gel, tooth cleaning powder, oral spray, etc. for maintaining oral health.

[0206] Preferred Example 1 Oral Care Composition The above oral preparation is prepared by adding 3 to 6 parts by weight.

[0207] Example 4 Preparation of Animal Oral Care Products

[0208] 1. Oral spray

[0209] Take 3-6 parts of the animal oral care composition of Example 1 of the present invention, 1-5 parts of a stabilizer, 1-5 parts of erythritol, 0.01-0.5 parts of a preservative, and the balance is deionized water.

[0210] The stabilizer is any one of propylene glycol and / or glycerol or a combination of both, and both are food grade.

[0211] The erythritol has a refreshing and pure sweet taste and is food grade.

[0212] The preservatives are sodium benzoate and / or potassium sorbate, both of which are food grade.

[0213] The components of the prepared samples are shown in the table:

[0214] Components (wt%) Sample 1 Sample 2 Sample 3 Animal oral care composition 3 5 6 Propylene glycol － － 5 glycerin 3 5 － Erythritol 4 3 5 Potassium sorbate 0.1 0.1 － Sodium benzoate － － 0.1 Deionized water margin margin margin total 100 100 100

[0215] Preparation method: dissolve a stabilizer, erythritol, and the oral care composition for animals of Example 1 in deionized water, then add a preservative, and stir to obtain the oral spray.

[0216] 2. Mouthwash

[0217] Take 3-6 parts of the animal oral care composition of Example 1 of the present invention, 0.1-0.5 parts of a thickener, 1-5 parts of a stabilizer, 1-5 parts of erythritol, 0.01-0.5 parts of a preservative, and the balance is deionized water.

[0218] The thickener is sodium carboxymethyl cellulose, which is food grade.

[0219] The stabilizer is propylene glycol and / or glycerol, both of which are food grade.

[0220] The erythritol has a refreshing and pure sweet taste and is food grade.

[0221] The preservatives are sodium benzoate and / or potassium sorbate, both of which are food grade.

[0222] The components of the prepared samples are shown in the table:

[0223]

[0224]

[0225] Preparation method: dissolve sodium carboxymethyl cellulose in deionized water, then dissolve a stabilizer, erythritol, and the oral care composition for animals of Example 1 in deionized water, and finally add a preservative and stir to obtain the mouthwash.

[0226] 3. Toothpaste

[0227] Take 3-6 parts of the animal oral care composition of Example 1 of the present invention, 0.5-2 parts of a thickener, 10-25 parts of an abrasive, 40-60 parts of a moisturizer, 0.01-0.5 parts of a preservative, 0.05-0.2 parts of a flavor, and the balance is deionized water.

[0228] The thickener is sodium carboxymethyl cellulose.

[0229] The friction agent is silicon dioxide.

[0230] The moisturizing agent is one or two of sorbitol, propylene glycol and glycerin.

[0231] The preservative is sodium benzoate and / or methylparaben

[0232] The flavor is food grade.

[0233] The components of the prepared samples are as follows:

[0234]

[0235]

[0236] Preparation method:

[0237] (1) dissolving the moisturizing agent in deionized water to form a solution;

[0238] (2) Mixing sodium carboxymethyl cellulose, silicon dioxide, and the oral care composition for animals in Example 1, adding the mixture to the above solution under vacuum, and stirring the mixture under vacuum to form a paste;

[0239] (3) Add essence and preservative to the above paste, stir evenly in vacuum to obtain the toothpaste.

[0240] 4. Tooth gel

[0241] Take 3-6 parts of the animal oral care composition of Example 1 of the present invention, 0.5-2 parts of a thickener, 1-5 parts of a moisturizer, 0.01-0.5 parts of a preservative, 0.05-0.2 parts of a flavor, and the balance is deionized water.

[0242] The thickener is sodium carboxymethyl cellulose.

[0243] The moisturizing agent is propylene glycol and / or glycerin.

[0244] The preservative is sodium benzoate and / or potassium sorbate.

[0245] The flavor is food grade.

[0246] The components of the prepared samples are as follows:

[0247] Components (wt%) Sample 1 Sample 2 Sample 3 Animal oral care composition 3 5 6 Sodium carboxymethyl cellulose 1.2 1.2 1.0 Propylene glycol － 3 3 glycerin 3 － 2 Potassium sorbate 0.1 0.1 0.1 essence 0.1 0.1 0.1 Deionized water margin margin margin total 100 100 100

[0248] Preparation method:

[0249] (1) dissolving sodium carboxymethyl cellulose in deionized water to form a solution;

[0250] (2) adding a moisturizer and the oral care composition for animals in Example 1 to the above solution and stirring uniformly to form a gel;

[0251] (3) Add essence and preservative to the gel and stir evenly to obtain the tooth gel.

[0252] This document uses specific examples to illustrate the principles and implementation methods of the present invention. The above examples are only intended to help understand the method and core concept of the present invention. At the same time, those skilled in the art will find that the specific implementation methods and application scopes may vary based on the concept of the present invention. In summary, the contents of this specification should not be construed as limiting the present invention.

Claims

1. An animal oral care composition, characterized in that The invention is composed of the following raw materials in parts by weight: 5 parts of persimmon extract, 0.5 parts of licorice chalcone A and 5 parts of gallnut extract; Persimmon extract is obtained from the fruit of the persimmon tree, of which persimmon tannin is the main component of persimmon extract; The purity of the licorice chalcone A is ≥95%; The gallnut extract is obtained by supercritical CO2 extraction.

2. An animal oral care composition according to claim 1, characterized in that The gallnut extract is a food-grade gallnut extract or a pharmaceutical-grade gallnut extract.

3. The method for preparing the animal oral care composition according to any one of claims 1 to 2, characterized in that: Here are the steps: The persimmon extract, licorice chalcone A and gallnut extract are mixed evenly to obtain the product.

4. The use of the animal oral care composition according to any one of claims 1 to 2, characterized in that: Used to prepare products for maintaining oral health in animals.

5. The use of the animal oral care composition according to claim 4, characterized in that: The product is one or more of oral spray, mouthwash, tooth cleaning powder, tooth gel and toothpaste.

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