Oral composition and application thereof

Abstract

The invention relates to an oral composition and application thereof, in particular to a composition which comprises N-acetylated breast milk oligosaccharide and fucosyllactose. Wherein the N-acetylated breast milk oligosaccharide is selected from the group consisting of lactose-N-tetraose and / or lactose-N-neotetraose, and the fucosyllactose is selected from the group consisting of 2 '-FL and / or 3-FL. The composition provided by the invention can improve the balance of microbial flora in the oral cavity so as to achieve the effects of improving and maintaining the health of the oral cavity.

Description

Technical Field

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

[0002] Oral health is an important part of overall health. In 2007, the World Health Organization proposed that oral disease is a serious public health problem that needs to be actively prevented and treated. Oral health includes: "no chronic pain in the oral and maxillofacial area, oropharyngeal cancer, oral ulcers, congenital defects such as cleft lip and palate, periodontal (gum) disease, caries, tooth loss, and other diseases and functional disorders that affect the oral cavity." Oral health directly or indirectly affects overall health. Oral diseases such as caries and periodontal disease can destroy the hard tissues of teeth and the supporting tissues around the teeth. In addition to affecting functions such as chewing, speech, and aesthetics, they can also cause difficulties in social interaction and psychological disorders. Some microorganisms exist in the mouth for a long time, which can cause or aggravate certain systemic diseases such as coronary heart disease and diabetes, endangering overall health and affecting the quality of life.

[0003] With the improvement of living standards, people pay more and more attention to oral health. However, oral problems such as oral ulcers, gingivitis, and periodontitis are still common, causing pain and discomfort to patients and affecting their quality of life. Although traditional oral treatment methods, such as drug therapy, can relieve symptoms to a certain extent, they often have problems such as local irritation, allergic reactions, repeated attacks, and large side effects. Therefore, it is particularly important to develop new, safe, and effective compositions that inhibit bacteria and reduce inflammation and improve oral ulcers. Summary of the invention

[0004] In order to overcome the deficiencies in the prior art, the present invention aims to provide a composition that is safe, significantly inhibits bacteria, and improves or cures oral ulcers.

[0005] In order to solve the above technical problems, the present invention adopts the following technical solutions:

[0006] The first object of the present invention is to provide a composition comprising N-acetylated human milk oligosaccharides and fucosyllactose;

[0007] Wherein, the N-acetylated human milk oligosaccharide is selected from lactose-N-tetraose and / or lactose-N-neotetraose, and the fucosyllactose is selected from 2'-FL and / or 3-FL.

[0008] Preferably, the composition consists of lactose-N-neotetraose and 2'-FL.

[0009] Preferably, the content of the N-acetylated human milk oligosaccharides is 30-70% by mass of the composition, and the remainder is fucosyllactose.

[0010] Preferably, the N-acetylated human milk oligosaccharides and fucosyllactose are each in the form of separate preparations.

[0011] Preferably, the N-acetylated human milk oligosaccharides and fucosyllactose are administered simultaneously or sequentially.

[0012] Preferably, the composition further comprises an auxiliary material.

[0013] The second object of the present invention is to provide a composition for improving oral health, wherein the composition for improving oral health comprises any one of the compositions described above.

[0014] Preferably, the content of the composition for improving oral health is 0.1-50% by mass.

[0015] The third object of the present invention is to provide use of any one of the above-mentioned compositions or the above-mentioned composition for improving oral health in the preparation of a product for improving oral health.

[0016] Preferably, the product is a product for improving and / or treating oral ulcers.

[0017] Preferably, the product is a product that inhibits the proliferation of oral pathogenic microorganisms.

[0018] Preferably, the oral pathogenic microorganisms include Porphyromonas gingivalis, Fusobacterium nucleatum and Streptococcus mutans.

[0019] Effects of the Invention

[0020] The components of the drug combination provided by the present invention are all natural ingredients, simple in composition, less in side effects, safe, reduced in use and dependence on artificial additives or other chemical components, and have high acceptance and compliance; the drug combination of the present invention can selectively promote the growth of beneficial bacteria (such as bifidobacteria, lactobacilli, etc.) in the oral cavity, while inhibiting the reproduction of harmful bacteria, thereby naturally regulating the balance of oral microecology and thus comprehensively improving oral health, thereby making up for the deficiency of traditional products that can only focus on solving a single problem. The components of the composition provided by the present invention show a significant synergistic effect in treating oral health diseases, and have significant application value and promotion prospects. DETAILED DESCRIPTION

[0021] In order to make the technical solutions and beneficial effects of the present invention more clearly understood, the following is a detailed description by listing specific embodiments. Unless otherwise defined, the technical and scientific terms used herein have the same meanings as those in the technical field to which this application belongs.

[0022] In this specification, the numerical range expressed using "a numerical value A to a numerical value B" means a range including the endpoints numerical values ​​A and B.

[0023] In the present specification, a numerical range expressed using "above" or "below" means a numerical range including the number.

[0024] In this specification, the word "may" means both performing a certain process and not performing a certain process.

[0025] In this specification, the unit names used are all international standard unit names, and unless otherwise stated, the "%" used indicates weight or mass percentage.

[0026] After extensive research, the inventors proposed a drug combination containing N-acetylated human milk oligosaccharides and fucosyllactose, specifically containing lactose-N-tetraose or lactose-N-neotetraose and 2'-FL or 3-FL. Furthermore, the inventors optimized the two components and found that when the two are combined in a certain ratio, the composition has excellent ability to regulate the balance of oral microecology, which helps to reduce the occurrence of oral diseases, and thus reduce the risk of systemic diseases caused by oral problems, such as cardiovascular diseases, diabetes, etc. A new drug ingredient is provided for oral care, oral soft tissue treatment or auxiliary treatment.

[0027] Lactose-N-tetraose (LNT) is composed of galactose, N-acetylglucosamine and glucose, accounting for about 6% (w / w) of total HMOs. LNT has promising physiological effects, including prebiotic properties, anti-adhesion, antibacterial activity and antiviral effects.

[0028] Lactose-N-neotetraose (LNnT) is one of the components with relatively high content in human milk oligosaccharides. It is a non-fucosylated neutral breast milk oligosaccharide with physiological functions such as immunomodulation, intestinal cell response regulator and promotion of cell maturation.

[0029] 2'-fucosyllactose (2'-FL) is the most abundant HMO (accounting for about 31%) and is an important immunologically active component in breast milk, playing an important role in the growth and development of infants and young children.

[0030] 3-fucosyllactose (3-FL) is an important component of human milk oligosaccharides, accounting for 25-55% of the total HMOs in 18-month breast milk. 3-FL plays an important role in promoting human health. For example, 3-fucosyllactose specifically inhibits the binding of pathogenic Escherichia coli and norovirus to the corresponding host receptors to avoid infection. 3-fucosyllactose can inhibit the interaction between Pseudomonas aeruginosa and local epithelial cells, thereby avoiding Pseudomonas aeruginosa infection of the gastrointestinal tract, urinary tract and respiratory system. 3-fucosyllactose can also specifically proliferate specific beneficial bacteria in the intestine to regulate the structure and function of the intestinal flora.

[0031] The compositions provided by the present invention are all human milk oligosaccharides (HMOs), which, as natural components in breast milk, have extremely high biocompatibility and can function naturally in the oral environment, reducing dependence on artificial additives or chemical components; HMOs can selectively promote the growth of beneficial bacteria in the oral cavity (such as Archaea Streptococcus salivarius, Bifidobacterium, Lactobacillus, etc.), while inhibiting the growth of harmful bacteria (such as Streptococcus mutans), thereby naturally regulating the balance of oral microecology, promoting saliva secretion, reducing inflammation, helping to reduce the occurrence of oral diseases, and thus reducing the risk of systemic diseases caused by oral problems, such as cardiovascular disease, diabetes, etc.; breast milk oligosaccharides also have the effect of enhancing the barrier function of the oral mucosa, reducing the invasion and damage of external pathogens to the oral mucosa, improving the self-protection ability of the oral cavity, and assisting in preventing the occurrence of caries; HMOs also have multiple health benefits such as regulating immunity, promoting brain development and regulating intestinal flora, which means that the use of oral health products containing HMOs may bring additional health benefits; at the same time, HMOs have extremely low toxicity, and when using oral disease treatment products containing the combination provided by the present invention, the user faces a smaller risk of side effects, making the subject more compliant and easier to accept. In summary, the composition provided by the present invention has significant advantages in solving the problems existing in other oral health products, including providing a more natural way to promote health, more comprehensive health benefits, fewer side effects and safety issues, higher acceptance and compliance, and making up for the deficiencies of traditional oral health products.

[0032] The first object of the present invention is to provide a composition comprising N-acetylated human milk oligosaccharides and fucosyllactose;

[0033] Wherein, the N-acetylated human milk oligosaccharide is selected from lactose-N-tetraose and / or lactose-N-neotetraose, and the fucosyllactose is selected from 2'-FL and / or 3-FL. The composition of the present invention can significantly improve oral health through multiple mechanisms such as inhibiting bacterial growth, promoting saliva secretion, improving oral flora balance, reducing inflammation, and assisting in preventing dental caries.

[0034] In the present application, N-acetylated human milk oligosaccharides and fucosyllactose are both in the form of powder particles. The present application does not limit the preparation process thereof, and any conventional process can be applied to the present application.

[0035] In certain embodiments, the composition consists of lactose-N-neotetraose and 2'-FL.

[0036] In certain embodiments, the content of N-acetylated human milk oligosaccharides is 30-70% (e.g., 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, etc.) based on the mass percentage of the composition, and the remainder is fucosyllactose.

[0037] In certain embodiments, the content of the N-acetylated human milk oligosaccharides is 50-70% by mass of the composition, and the remainder is fucosyllactose.

[0038] In certain embodiments, the N-acetylated human milk oligosaccharides and fucosyllactose are each in the form of separate formulations.

[0039] In certain embodiments, the N-acetylated human milk oligosaccharides and fucosyllactose are administered simultaneously or sequentially.

[0040] In certain embodiments, the composition further comprises an excipient.

[0041] In certain embodiments, the excipients may be appropriate solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, adhesives, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adhesives, integrities, penetration enhancers, pH regulators, buffers, plasticizers, surfactants, foaming agents, defoamers, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants and deflocculating agents, filter aids, release retardants, etc.

[0042] In certain embodiments, the composition of the present application can be prepared by a general method, wherein one or more diluents or carriers can be added, for example, in an oral form, such as a gel, tablet, capsule, granule, powder, lozenge, syrup, emulsion, suspension, etc.

[0043] In certain embodiments, the composition can also be compounded with known effective ingredients.

[0044] The second object of the present invention is to provide a composition for improving oral health, wherein the composition for improving oral health comprises any one of the above-mentioned compositions. Oral health products (such as toothpaste, mouthwash, etc.) containing the composition of the present invention are easy to use, help improve user compliance, and thus ensure the continued effect of the product.

[0045] In certain embodiments, the content of the composition for improving oral health is 0.1-50%, for example, 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, etc., measured by mass percentage of the composition.

[0046] The third object of the present invention is to provide use of any one of the above-mentioned compositions or the above-mentioned composition for improving oral health in the preparation of a product for improving oral health.

[0047] In certain embodiments, the product is a product for improving and / or treating oral ulcers.

[0048] In certain embodiments, the product is a product that inhibits the proliferation of oral pathogenic microorganisms.

[0049] In certain embodiments, the oral pathogenic microorganisms include Porphyromonas gingivalis, Fusobacterium nucleatum, and Streptococcus mutans.

[0050] In a preferred embodiment, the oral health improving product may be an ointment, a mouthwash, a toothpaste, a tooth powder, and the like.

[0051] The present invention will be further described below in conjunction with specific embodiment.It should be appreciated that these embodiments are only used to illustrate the present invention and are not intended to limit the scope of the present invention.The conditions of the experimental techniques not specifically described in the following examples are usually according to the normal conditions described in Sambrook et al., Molecular Cloning:A Laboratory Manual (New York:Cold Spring Harbor Laboratory Press, 1989), or according to the conditions described in Microbiology Magazine: Laboratory Manual (edited by James Cappuccino and Natalie Sherman of Pearson Education Press) or the conditions recommended by manufacturers.Unless otherwise indicated, percentages and parts are weight percentages and parts by weight.

[0052] Unless otherwise stated, the materials used in the examples are commercially available products.

[0053] Example 1

[0054] 300 people with oral ulcers were recruited and divided into 6 groups, 50 people in each group. The formula powder in Table 1 was dissolved in 50 mL of sterile water. The patients rinsed their mouths once every morning and evening after meals for 5 consecutive days, and the healing status on the 6th day was recorded. The evaluation criteria were that the ulcer base gradually flattened, the ulcer gradually shrank or disappeared, the mucosal congestion was significantly reduced, the inflammation subsided, and the pain was significantly improved or disappeared. The oral ulcer healing efficiency in Table 1 = the number of people judged to be effective / 50*100%.

[0055] Table 1

[0056] Combination / single product Ratio (total amount 10g) Oral ulcer healing efficiency (5 days) Z1 2'-FL30%+LNnT70% 94％ Z2 2'-FL50%+LNnT50% 90％ Z3 2'-FL70%+LNnT30% 86％ Z4 3-FL30%+LNnT70% 90％ Z5 3-FL50%+LNnT50% 84％ Z6 3-FL70%+LNnT30% 82％ Control group starch 66％ D1 2'-FL 80％ D2 LqCy 82％ D3 3-FL 78％

[0057] It can be seen from Table 1 that, compared with the control group and groups D1 to D3, the compositions of groups Z1 to Z6 have the efficacy of promoting the healing of oral ulcers, especially the composition of group Z1 has the strongest ability to promote the healing of oral ulcers, reaching 94%.

[0058] Example 2

[0059] Streptococcus, as oral planktonic bacteria, colonizes on the tooth surface in the early stage. Streptococcus mutans is the main microorganism that causes caries. It can ferment the residual carbohydrates in the mouth to produce acid and form a layer of adhesive plaque on the tooth surface, so that the produced organic acid contacts the tooth surface for a long time, causing damage to the tooth hard tissue, forming caries holes, and affecting oral health. Porphyromonas gingivalis is a late colonizing bacterium, and Fusobacterium nucleatum can connect the two. Porphyromonas gingivalis and Fusobacterium nucleatum play a leading role in periodontal disease. This example verifies that the composition of the present invention has an inhibitory effect on the proliferation of the above three microorganisms, thereby further illustrating that the combination of the present application is beneficial to oral health.

[0060] 1. Culture medium: Porphyromonas gingivalis, Fusobacterium nucleatum and other suspected periodontal pathogens need to be cultured anaerobically at 37°C in BHI blood broth medium under a mixed gas of 80% N2, 10% H2 and 10% CO2; oral Streptococcus (Streptococcus mutans) needs to be cultured statically in BHI medium at 37°C with 5% CO2 or under anaerobic conditions.

[0061] 2. Equipment

[0062] Constant temperature water bath, timer, Class II biological safety cabinet, etc.

[0063] 3. Specific steps

[0064] (1) Bacterial culture: The primary revived Porphyromonas gingivalis and Fusobacterium nucleatum were transferred to BHI liquid medium and cultured at 37°C under anaerobic conditions (80% N2, 10% H2 and 10% CO2) for 48 h. After three generations, the strains reached a stable state and were used for subsequent experiments. The primary revived Streptococcus mutans were transferred to BHI liquid medium and cultured at 37°C under anaerobic conditions (5% CO2) for 48 h. After three generations, the strains reached a stable state and were used for subsequent experiments.

[0065] (2) Preparation of culture medium: Take 10 mL of the composition / single product of Example 1 (excluding starch) and 10 mL of distilled water as a control group, add 3.6 g of BHI culture medium powder, 0.5 mL of 5% sterile defibrinated sheep blood, 50 μL of hemin chloride, and 50 μL of vitamin K1, dissolve and filter with a 0.22 μm filter membrane to sterilize to prepare a liquid culture medium for use.

[0066] (3) Inoculate 2% volume fraction of the activated pathogenic bacteria in the culture medium prepared in step 2 and culture them in a screw anaerobic tube at 37°C for 48h. Take the cultured bacterial solution and spread it on a BHI blood plate by plate dilution coating method, and invert the blood plate under anaerobic conditions at 37°C (Porphyromonas gingivalis, Fusobacterium nucleatum: 80% N2, 10% H2 and 10% CO2; Streptococcus mutans: 5% CO2) for 24h, and observe and count the number of live bacteria. The experiment was repeated three times independently. The counting principle was based on the national standard GB4789.2-2016. The colony count was selected between 30CFU and 300CFU for colony counting, and the results were expressed as log10CFU / mL. The antibacterial rate of each embodiment and comparative example against pathogenic bacteria was calculated, and the calculation formula was: antibacterial rate = (number of live bacteria in the blank control - number of live bacteria in the experimental group) / number of live bacteria in the blank control × 100%.

[0067] The results showed that after three repeated tests, the test samples 1-6 were exposed to Streptococcus mutans, Porphyromonas gingivalis, and Fusobacterium nucleatum for 1 hour, and the antibacterial rate test results were shown in Table 2. In Table 2, if the antibacterial rate was ≥50% to 90%, it was judged to have an antibacterial effect; if the antibacterial rate was ≥90%, it was judged to have a strong antibacterial effect.

[0068] Table 2 Antibacterial test results of samples

[0069]

[0070]

[0071] As can be seen from Table 2, different compositions have different inhibitory effects on pathogenic bacteria. Composition 1 has a strong inhibitory effect on Streptococcus mutans and Fusobacterium nucleatum, especially inhibiting Streptococcus mutans, and has an antibacterial effect on Porphyromonas gingivalis; Composition 2 has a strong inhibitory effect on Streptococcus mutans and Fusobacterium nucleatum, and has an antibacterial effect on Porphyromonas gingivalis; Composition 3 has a strong antibacterial effect on all three pathogenic bacteria; Composition 4 has a strong inhibitory effect only on Fusobacterium nucleatum, and has an inhibitory effect on Streptococcus mutans and Porphyromonas gingivalis; Combinations 5 and 6 and the three single products have an antibacterial effect on the three pathogenic bacteria. In summary, the composition of the present invention has an inhibitory effect on the proliferation of the above three microorganisms, improves the balance of oral flora, thereby improving and maintaining oral health, and reducing the risk of caries and other oral diseases.

[0072] It should be understood that the above embodiments are exemplary and are not intended to include all possible implementations included in the claims. Various modifications and changes may be made on the basis of the above embodiments without departing from the scope of the present disclosure. Similarly, the various technical features of the above embodiments may be arbitrarily combined to form other embodiments of the present invention that may not be explicitly described. Therefore, the above embodiments only express several implementations of the present invention and do not limit the scope of protection of the patent of the present invention.

Claims

1. A composition, characterized in that The composition comprises N-acetylated human milk oligosaccharides and fucosyllactose; Wherein, the N-acetylated human milk oligosaccharide is selected from lactose-N-tetraose and / or lactose-N-neotetraose, and the fucosyllactose is selected from 2'-FL and / or 3-FL.

2. The composition according to claim 1, characterized in that The composition consists of lactose-N-neotetraose and 2'-FL.

3. The composition according to claim 1, characterized in that Calculated by mass percentage of the composition, the content of the N-acetylated human milk oligosaccharide is 30-70%, and the remainder is fucosyllactose.

4. The composition according to any one of claims 1 to 3, characterized in that The N-acetylated human milk oligosaccharides and fucosyllactose are each in the form of separate preparations.

5. The composition according to claim 4, characterized in that The N-acetylated human milk oligosaccharides and fucosyllactose are administered simultaneously or sequentially.

6. The composition according to claim 1, characterized in that The composition further includes auxiliary materials.

7. A composition for improving oral health, characterized in that: The composition for improving oral health comprises the composition according to any one of claims 1 to 6.

8. The composition for improving oral health according to claim 7, characterized in that: Calculated by the mass percentage of the composition for improving oral health, the content of the composition is 0.1-50%.

9. Use of the composition according to any one of claims 1 to 6 or the composition for improving oral health according to claim 7 or 8 in the preparation of a product for improving oral health.

10. The use according to claim 9, characterized in that: The product is a product for improving and / or treating oral ulcers; Preferably, the product is a product that inhibits the proliferation of oral pathogenic microorganisms; Preferably, the oral pathogenic microorganisms include Porphyromonas gingivalis, Fusobacterium nucleatum and Streptococcus mutans.

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