Biological peptide antibacterial mouthwash as well as preparation method and application thereof

Abstract

The invention discloses biological peptide antibacterial mouthwash as well as a preparation method and application thereof. The mouthwash comprises small molecule peptides shown as SEQ NO.1, SEQ NO.2 and SEQ NO.3 in a specific proportion, and auxiliaries such as glycerol, sodium hyaluronate and cocamidopropyl betaine. The small molecule peptides play roles in oral bacterial cell wall synthesis, cell membrane transport and cell membrane energy metabolism related proteins respectively, and a synergistic antibacterial effect is generated after combination. According to the present invention, the inhibition zone test and the minimum inhibitory concentration determination results show that the antibacterial effect of the mouthwash on streptococcus mutans, porphyromonas gingivalis and fusobacterium nucleatum is significantly superior to the traditional mouthwash and the single biological peptide mouthwash, and the mouthwash has advantages of good biocompatibility, low toxicity, effective oral cavity cleaning and oral cavity disease prevention, and is suitable for daily oral cavity nursing.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine and chemistry, and specifically relates to a biopeptide antibacterial mouthwash and a preparation method and application thereof. Background Art

[0002] The oral cavity is a complex micro-ecosystem where numerous microorganisms live. Under normal circumstances, oral microorganisms are in balance with the human body, but when the balance is broken, various oral diseases will occur. Problems such as caries, gingivitis, periodontitis and halitosis plague many people and seriously affect their quality of life.

[0003] Traditional mouthwashes mostly use chemical bactericides such as chlorhexidine as the main antibacterial ingredients. Although chlorhexidine can inhibit the growth of oral microorganisms, its long-term use has many disadvantages. It can easily cause irritation of the oral mucosa, causing discomfort to the user, and may even cause damage to the oral mucosa. Moreover, chlorhexidine can also stain teeth, affecting the appearance of teeth, which is unacceptable for users who pay attention to their image. In addition, with the improvement of people's health awareness, the demand for natural and safe oral care products is becoming more and more urgent.

[0004] Biopeptides have gradually become a research hotspot in the field of oral care due to their good biocompatibility, low toxicity and unique biological activity. However, existing biopeptide mouthwashes still have some shortcomings. The antibacterial effect of some biopeptide mouthwashes is not ideal, and it is difficult to effectively deal with a variety of oral pathogens. The stability of some biopeptide mouthwashes is poor, and the active ingredients are easily inactivated or degraded during storage and use, thereby affecting the antibacterial performance and shelf life of the product. Moreover, a single biopeptide can often only act on a specific target or physiological process of bacteria, and the antibacterial spectrum is relatively narrow, making it difficult to achieve a comprehensive and efficient antibacterial effect.

[0005] Based on the above background, it is of great practical significance to develop a biopeptide antibacterial mouthwash with high antibacterial performance, good stability, safety and non-irritation. The present invention aims to overcome the defects of the prior art by designing a new small molecule peptide sequence and optimizing the mouthwash formula, and provide a better quality product for the oral care market. Summary of the invention

[0006] The present invention provides a biopeptide antibacterial mouthwash comprising the following small molecule peptide SEQ NO.1: Leu-His-Pro-Gly-Tyr-Lys-Glu.

[0007] Furthermore, it also contains the following small molecule peptide SEQ NO.2: Arg-Trp-Ser-Ile-Asp-Met-Cys.

[0008] Furthermore, it also contains the following small molecule peptide SEQ NO6: Phe-Ala-Val-Gln-Thr-Asn-Leu. On the other hand, a biological peptide antibacterial mouthwash contains the following ingredients in weight percentage: SEQ NO.1 (Leu-His-Pro-Gly-Tyr-Lys-Glu) small molecule peptide: 0.15 parts to 0.35 parts;

[0009] SEQ NO.2 (Arg-Trp-Ser-Ile-Asp-Met-Cys) small molecule peptide: 0.1-0.2 parts;

[0010] SEQ NO.3 (Phe-Ala-Val-Gln-Thr-Asn-Leu) small molecule peptide: 0.1-0.2 parts;

[0011] Moisturizer: 5-10 parts of glycerin and 0.1-0.3 parts of sodium hyaluronate;

[0012] Surfactant: 1-3 parts of cocamidopropyl betaine;

[0013] Flavoring: 0.1-0.3 parts of menthol and 2-5 parts of xylitol;

[0014] Preservative: potassium sorbate 0.05-0.15 parts;

[0015] Buffer: 0.2-0.5 parts of disodium hydrogen phosphate and 0.1-0.3 parts of citric acid;

[0016] Slow-release organic compound: ethyl 2-(2-hydroxyphenoxy)acetate, 1 to 3 parts.

[0017] The rest was deionized water.

[0018] Furthermore, the small molecule peptide of SEQ NO. 1 specifically binds to the active site of the transglycosylase in the synthesis of the oral bacterial cell wall, thereby hindering the extension of the sugar chain in the process of peptidoglycan synthesis, thereby inhibiting bacterial growth.

[0019] Furthermore, the SEQ NO. 2 small molecule peptide interacts with the substrate binding site of the ATP-binding cassette transporter on the cell membrane of oral bacteria, thereby preventing the uptake of nutrients and the discharge of harmful substances, thereby inhibiting bacterial proliferation.

[0020] Furthermore, the SEQ NO.3 small molecule peptide binds to the proton motive force-related protein on the oral bacterial cell membrane, interferes with its normal conformation and function, destroys the energy conversion and material transport balance of the bacterial cell membrane, and thus inhibits bacterial growth.

[0021] Further, when SEQ NO.1, SEQ NO.2 and SEQ NO.3 are combined in a ratio of 3:2:2, they have a synergistic antibacterial effect on a mixed flora of Streptococcus mutans, Porphyromonas gingivalis and Fusobacterium nucleatum. When the total peptide concentration is 0.3 mg / mL, the inhibition rate of the mixed flora is more than 85 parts. Further, sustained-release organic compound: 2-(2-hydroxyphenoxy)ethyl acetate, 1 part to 3 parts. 2-(2-hydroxyphenoxy)ethyl acetate This compound has an ester group and a hydroxyphenoxy structure. The ester group can be slowly hydrolyzed in the oral environment to release hydroxyphenoxyacetic acid with antibacterial activity. Hydroxyphenoxyacetic acid can react with amino groups, sulfhydryl groups and other groups in the protein and enzyme molecules of various oral bacteria to change their structure and function, thereby enhancing the antibacterial effect of the mouthwash.

[0022] On the other hand, the present invention also provides a method for preparing the biopeptide antibacterial mouthwash, comprising the following steps:

[0023] S1 uses chemical synthesis to prepare SEQ NO.1, SEQ NO.2 and SEQ NO.3 small molecule peptides. During the synthesis process, a suitable solid phase synthetic resin is selected, and the corresponding amino acids are modified with protective groups, and then the amino acids are coupled to the resin in sequence. The reaction temperature is controlled at 20-30°C, the pH value is 6-8, and the coupling time of each amino acid is 1-2 hours. After the peptide chain is synthesized, the protective group is removed and cleaved, and then purified and identified by high performance liquid chromatography, so that the purity of the peptide reaches more than 95 parts;

[0024] S2: mixing the purified SEQ NO.1, SEQ NO.2 and SEQ NO.3 small molecule peptides with a moisturizer, a surfactant, a flavoring agent, a preservative and a buffer in an appropriate amount of deionized water according to the ratio of claim 1, first dissolving the small molecule peptides in a small amount of deionized water, then adding other ingredients in sequence, and slowly stirring at a speed of 200-400 rpm in a stirrer for 30-60 minutes until uniform;

[0025] S3 finally filters the mixed solution with a 0.22 μm microporous filter membrane to remove impurities, obtains the finished product of the bio-peptide antibacterial mouthwash, and performs aseptic packaging.

[0026] The application of the biopeptide antibacterial mouthwash in oral care is used to prevent and improve oral diseases caused by Streptococcus mutans, Porphyromonas gingivalis and Fusobacterium nucleatum, including caries, gingivitis and halitosis. Beneficial effects of the invention:

[0027] The biopeptide antibacterial mouthwash of the present invention plays a role in multiple key targets of oral bacteria by containing three new small molecule peptides, SEQ NO.1, SEQ NO.2 and SEQ NO.3. The combination of the three small molecule peptides produces a synergistic effect, not a simple superposition. SEQ NO.1 destroys the bacterial cell wall structure, creating conditions for SEQ NO.2 and SEQ NO.3 to bind to the cell membrane target; SEQ NO.2 interferes with the function of cell membrane transport proteins, weakens the metabolism and repair ability of bacteria, and acts on the cell membrane system together with SEQ NO.3; SEQ NO.3 destroys the energy and material transport balance of bacterial cell membranes, which in turn affects the response of bacteria to cell wall damage and the maintenance of transport protein function. This synergistic effect attacks bacteria from multiple aspects and greatly enhances the antibacterial effect. The biopeptide is composed of natural amino acids, has good biocompatibility, and is extremely irritating to oral mucosa and teeth. Unlike traditional mouthwashes containing chemical bactericides, the mouthwash of the present invention reduces the potential risks of oral mucosal damage, tooth staining, etc. caused by component stimulation, and is suitable for long-term use. Especially for children, pregnant women and people with oral sensitivity, it provides a safe and reliable oral care option that can effectively prevent and treat oral diseases while protecting the health of users. In addition to the biopeptide ingredients, mouthwash also contains a variety of carefully selected adjuvants. Moisturizers glycerin and sodium hyaluronate can keep the mouth moist, prevent dryness and discomfort, and help maintain product stability; surfactant cocamidopropyl betaine can reduce surface tension, allowing the mouthwash to better penetrate and clean various parts of the mouth, and improve the antibacterial effect; flavoring agents menthol and xylitol give fresh breath and good taste, and xylitol has an anti-caries effect; preservative potassium sorbate ensures product quality and safety and prevents microbial contamination; buffers disodium hydrogen phosphate and citric acid maintain a suitable pH value to ensure the activity of biopeptides and oral health. 2-(2-hydroxyphenoxy)ethyl acetate (HPAE) in the mouthwash formula is a slow-release organic compound that can effectively prolong the antibacterial time of the mouthwash. The ester group of HPAE slowly hydrolyzes in the oral environment, continuously releasing hydroxyphenoxyacetic acid with antibacterial activity, which synergizes with biopeptides to continuously inhibit the growth and reproduction of oral bacteria. Compared with many existing mouthwashes, the antibacterial effect of the mouthwash of the present invention can last longer, can more reliably protect oral health, and reduce the risk of oral bacteria re-growth due to short-term antibacterial effect. The various components work together to make the mouthwash not only perform well in antibacterial performance, but also comprehensively improve the oral health and comfort of users. It has strong competitiveness in the oral care product market and provides a comprehensive and excellent solution for oral care. DETAILED DESCRIPTION

[0028] Example 1

[0029] The purpose of this experiment is to compare the differences in antibacterial effects between the bio-peptide antibacterial mouthwash of the present invention (including a novel small molecule peptide combination) and existing traditional mouthwashes and single bio-peptide mouthwashes, so as to highlight the synergistic advantages of the present invention.

[0030] The mouthwash of the present invention is a biological peptide antibacterial mouthwash prepared according to the preparation method described in the above invention content, wherein SEQ NO.1 (Leu-His-Pro-Gly-Tyr-Lys-Glu), SEQ NO.2 (Arg-Trp-Ser-Ile-Asp-Met-Cys) and SEQ NO.3 (Phe-Ala-Val-Gln-Thr-Asn-Leu) small molecule peptides are combined in a ratio of 3:2:2, and the content of each component is within a specified range, and the total peptide concentration is 0.3 mg / mL.

[0031] Traditional mouthwash: A representative traditional mouthwash containing chlorhexidine on the market was selected, with a chlorhexidine concentration of 0.12 parts (w / v), which is a common effective antibacterial concentration range.

[0032] Single biological peptide mouthwash 1: a mouthwash containing only the SEQ NO.1 small molecule peptide, with a peptide concentration of 0.2 mg / mL. Other ingredients are similar to the mouthwash of the present invention, but do not contain SEQ NO.2 and SEQ NO.3 peptides and the proportion of ingredients is adjusted accordingly.

[0033] Single biological peptide mouthwash 2: A mouthwash containing only SEQ NO.2 small molecule peptide, with a peptide concentration of 0.15 mg / mL and the same ingredient adjustment.

[0034] Single biological peptide mouthwash 3: A mouthwash containing only SEQ NO.3 small molecule peptide, with a peptide concentration of 0.18 mg / mL and the same ingredient adjustment.

[0035] Test strains: Streptococcus mutans, Porphyromonas gingivalis and Fusobacterium nucleatum, all purchased from professional microbial strain collection institutions, were revived, cultured and adjusted to the logarithmic growth phase before the test.

[0036] Test method: Prepare a Müller-Hinton agar medium plate and divide the plate into several areas. Take 10 μL of different mouthwash samples and drop them on the surface of the medium in the corresponding area. Place an Oxford cup (diameter 8 mm) to spread the sample evenly. Use a sterile cotton swab to dip an appropriate amount of the test strain suspension (concentration 1×10 7CFU / mL), spread evenly on the plate surface, and culture at 37°C for 24 hours. Measure the diameter of the inhibition zone, repeat 3 times for each sample, and take the average value.

[0037] Using the liquid dilution method, different mouthwashes were diluted with sterile culture medium in a 96-well microplate, with a final volume of 200 μL per well. 20 μL of the test strain suspension (concentration of 1×10 6 CFU / mL), and cultured at 37°C for 48 hours. Observe the turbidity of the liquid in the well, and take the lowest mouthwash concentration without turbidity growth as the MIC of the sample for the corresponding strain. Repeat each sample 3 times.

[0038] Test results

[0039] (I) Results of inhibition zone test

[0040] 1. Antibacterial effect on Streptococcus mutans

[0041] 1. The average diameter of the antibacterial zone of the mouthwash of the present invention is 22.5 mm.

[0042] 2. The average diameter of the antibacterial zone of traditional mouthwash is 18.2mm.

[0043] 3. The average diameter of the inhibition zone of single biopeptide mouthwash 1 was 13.5 mm.

[0044] 4. The average diameter of the inhibition zone of single biopeptide mouthwash 2 was 11.8 mm.

[0045] 5. The average diameter of the antibacterial zone of single biopeptide mouthwash 3 was 12.6 mm. 2. Antibacterial effect on Porphyromonas gingivalis

[0046] 1. The average diameter of the antibacterial zone of the mouthwash of the present invention is 20.8 mm.

[0047] 2. The average diameter of the antibacterial zone of traditional mouthwash is 16.5mm.

[0048] 3. The average diameter of the inhibition zone of single biopeptide mouthwash 1 was 10.2 mm.

[0049] 4. The average diameter of the inhibition zone of single biopeptide mouthwash 2 was 14.3 mm.

[0050] 5. The average diameter of the inhibition zone of single biopeptide mouthwash 3 was 11.5 mm.

[0051] 3. Antibacterial effect on Fusobacterium nucleatum

[0052] 1. The average diameter of the antibacterial zone of the mouthwash of the present invention is 21.6 mm.

[0053] 2. The average diameter of the antibacterial zone of traditional mouthwash is 17.3mm.

[0054] 3. The average diameter of the inhibition zone of single biopeptide mouthwash 1 was 11.9 mm.

[0055] 4. The average diameter of the inhibition zone of single biopeptide mouthwash 2 was 10.8 mm.

[0056] 5. The average diameter of the antibacterial zone of single biopeptide mouthwash 3 was 13.2 mm. (II) Minimum inhibitory concentration (MIC) test results

[0057] 1. MIC against Streptococcus mutans

[0058] 1. The MIC of the mouthwash of the present invention is 0.03 mg / mL.

[0059] 2. The MIC of traditional mouthwash is 0.06 parts (w / v), which is approximately 0.6 mg / mL after conversion.

[0060] 3. The MIC of single biopeptide mouthwash 1 is 0.2 mg / mL.

[0061] 4. The MIC of single biopeptide mouthwash 2 was 0.3 mg / mL.

[0062] 5. The MIC of single biopeptide mouthwash 3 was 0.25 mg / mL.

[0063] 2. MIC against Porphyromonas gingivalis

[0064] 1. The MIC of the mouthwash of the present invention is 0.04 mg / mL.

[0065] 2. The MIC of traditional mouthwash is 0.08 parts (w / v), which is about 0.8 mg / mL.

[0066] 3. The MIC of single biopeptide mouthwash 1 was 0.4 mg / mL.

[0067] 4. The MIC of single biopeptide mouthwash 2 was 0.2 mg / mL.

[0068] 5. The MIC of single biopeptide mouthwash 3 was 0.3 mg / mL.

[0069] 3. MIC against Fusobacterium nucleatum

[0070] 1. The MIC of the mouthwash of the present invention is 0.035 mg / mL.

[0071] 2. The MIC of traditional mouthwash is 0.07 parts (w / v), which is about 0.7 mg / mL.

[0072] 3. The MIC of single biopeptide mouthwash 1 was 0.3 mg / mL.

[0073] 4. The MIC of single biopeptide mouthwash 2 was 0.4 mg / mL.

[0074] 5. The MIC of single biopeptide mouthwash 3 was 0.2 mg / mL.

[0075] From the results of the inhibition zone test, the diameters of the inhibition zones of the mouthwash of the present invention for the three test strains are significantly larger than those of the traditional mouthwash. This shows that the mouthwash of the present invention has a more excellent performance in inhibiting the spread of bacterial growth and can prevent the growth and reproduction of bacteria in a larger area. In the MIC determination results, the MIC values ​​of the mouthwash of the present invention for Streptococcus mutans, Porphyromonas gingivalis and Fusobacterium nucleatum are much lower than those of the traditional mouthwash. This means that the mouthwash of the present invention can effectively inhibit the growth of these bacteria at a lower concentration, indicating that its antibacterial activity is stronger, and because a lower concentration of active ingredients is used, the potential side effects caused by high concentrations of chemical bactericides can be reduced, such as oral mucosal irritation, tooth staining and other problems.

[0076] In the inhibition zone test, the diameter of the inhibition zone of the mouthwash of the present invention for each strain is greater than that of the single biological peptide mouthwash. This shows that the combination of the three new small molecule peptides produces a synergistic effect, and its antibacterial effect is not a simple superposition of a single peptide, but cooperates with each other through a multi-target mechanism of action, expanding the range and intensity of antibacterial. In terms of MIC determination, the MIC value of the mouthwash of the present invention is significantly lower than that of a single biological peptide mouthwash. For example, for Streptococcus mutans, the MIC of the single biological peptide mouthwash 1 is 0.2 mg / mL, while the mouthwash of the present invention only requires 0.03 mg / mL, which fully proves that the combination of the three peptides greatly reduces the minimum effective concentration required to inhibit bacterial growth, reflecting a strong synergistic effect, and further illustrates that the mouthwash of the present invention optimizes the antibacterial performance through a reasonable peptide combination design, and has obvious advantages and innovations in the field of oral care products.

[0077] Example 2

[0078] This experiment aims to study the synergistic effect of 2-(2-hydroxyphenoxy)ethyl acetate (HPAE) in biological peptide antibacterial mouthwash, and compare it with the mouthwash without HPAE to evaluate its effect on the antibacterial effect and durability. Experimental mouthwash 1: The biological peptide antibacterial mouthwash prepared according to the preparation method described in the above invention content contains SEQ NO.1, SEQ NO.2 and SEQ NO.3 small molecule peptides, glycerol, sorbitol, sodium laureth sulfate, natural menthol, sweet orange flavor, sodium benzoate, HPAE and deionized water, and the content of each component is within the specified range.

[0079] Test mouthwash 2: Similar to test mouthwash 1, but without HPAE, and the contents of other ingredients were adjusted accordingly. Test strains: Streptococcus mutans, Porphyromonas gingivalis, and Fusobacterium nucleatum, all purchased from professional microbial strain collection institutions, revived, cultured, and adjusted to the logarithmic growth phase before testing.

[0080] Prepare a Müller-Hinton agar medium plate and divide the plate into several areas. Take 10 μL of different mouthwash samples and drop them on the surface of the medium in the corresponding area. Place an Oxford cup (diameter 8 mm) to spread the sample evenly. Use a sterile cotton swab to dip an appropriate amount of the test strain suspension (concentration 1×10 7 CFU / mL), spread evenly on the plate surface, and culture at 37°C for 24 hours. Measure the diameter of the inhibition zone, repeat 3 times for each sample, and take the average value.

[0081] Using the liquid dilution method, different mouthwashes were diluted with sterile culture medium in a 96-well microplate, with a final volume of 200 μL per well. 20 μL of the test strain suspension (concentration of 1×10 6 CFU / mL), and cultured at 37°C for 48 hours. Observe the turbidity of the liquid in the well, and take the lowest mouthwash concentration without turbidity growth as the MIC of the sample for the corresponding strain. Repeat each sample 3 times.

[0082] Mouthwash 1 and mouthwash 2 were mixed with bacterial suspensions of Streptococcus mutans, Porphyromonas gingivalis, and Fusobacterium nucleatum (concentration of 1×10 6 CFU / mL) and incubate at 37°C. Take a small amount of the mixture every 2 hours and inoculate it on the corresponding culture medium plate. After culturing for 24 hours, observe the growth of the colony to determine the duration of the antibacterial effect of the mouthwash.

[0083] Test results: The average diameter of the antibacterial zone of the test mouthwash 1 (containing HPAE) against Streptococcus mutans was 23.8 mm.

[0084] The average diameter of the inhibition zone of the test mouthwash 2 (without HPAE) was 22.5 mm.

[0085] The average diameter of the inhibition zone of the test mouthwash 1 (containing HPAE) was 21.6 mm. The average diameter of the inhibition zone of the test mouthwash 2 (without HPAE) was 20.8 mm. For the antibacterial effect on Fusobacterium nucleatum, the average diameter of the inhibition zone of the test mouthwash 1 (containing HPAE) was 22.9 mm. The average diameter of the inhibition zone of the test mouthwash 2 (without HPAE) was 21.6 mm.

[0086] The results of MIC determination showed that the MIC of test mouthwash 1 (containing HPAE) against mutans Streptococcus was 0.025 mg / mL. The MIC of test mouthwash 2 (without HPAE) was 0.03 mg / mL. The MIC of test mouthwash 1 (containing HPAE) against Porphyromonas gingivalis was 0.035 mg / mL. The MIC of test mouthwash 2 (without HPAE) was 0.04 mg / mL. The MIC of test mouthwash 1 (containing HPAE) against Fusobacterium nucleatum was 0.03 mg / mL. The MIC of test mouthwash 2 (without HPAE) was 0.035 mg / mL. The results of durability test showed that the antibacterial effect of test mouthwash 1 (containing HPAE) against mutans Streptococcus could last for more than 8 hours; against Porphyromonas gingivalis could last for more than 7 hours; and against Fusobacterium nucleatum could last for more than 7.5 hours. The antibacterial effect of the experimental mouthwash 2 (without HPAE) on Streptococcus mutans can last for about 6 hours; on Porphyromonas gingivalis can last for about 5.5 hours; and on Fusobacterium nucleatum can last for about 6 hours.

[0087] Slow-release organic compound: 2-(2-hydroxyphenoxy)ethyl acetate (hereinafter referred to as HPAE), with a structural formula of . This compound has an ester group and a hydroxyphenoxy structure. The ester group can be slowly hydrolyzed in the oral environment to release hydroxyphenoxyacetic acid with antibacterial activity. Hydroxyphenoxyacetic acid can react with amino groups, sulfhydryl groups and other groups in the proteins and enzyme molecules of various oral bacteria, changing their structure and function, thereby enhancing the antibacterial effect of mouthwash.

[0088] It can be seen from the test results that 2-(2-hydroxyphenoxy)ethyl acetate (HPAE) has a significant synergistic effect in biological peptide antibacterial mouthwashes. In the antibacterial zone test, the diameter of the antibacterial zone of the mouthwash containing HPAE for the three test strains was larger than that of the mouthwash without HPAE, indicating that HPAE can enhance the antibacterial ability of mouthwash. In the MIC determination results, the MIC values ​​of the mouthwash containing HPAE for each strain were lower than those of the mouthwash without HPAE, indicating that HPAE can reduce the minimum effective concentration required for mouthwash to inhibit bacterial growth. In terms of durability test, the antibacterial effect of the mouthwash containing HPAE lasted significantly longer than that of the mouthwash without HPAE. This is because the ester group of HPAE is slowly hydrolyzed in the oral environment, and the released hydroxyphenoxyacetic acid can react with the protein and enzyme molecules of oral bacteria, enhancing the antibacterial effect of the biological peptide, thereby improving the antibacterial effect and durability of the mouthwash.

[0089] In summary, compared with the prior art, the bio-peptide antibacterial mouthwash of the present invention, whether compared with traditional mouthwash or single bio-peptide mouthwash, shows a significant synergistic effect in antibacterial effect, has better application prospects and market potential, and is expected to provide a safer and more efficient solution for oral care.

Claims

1. A biopeptide antibacterial mouthwash, characterized in that: The small molecule peptides specifically bind to the active site of the transglycosylase in the synthesis of the oral bacterial cell wall, hindering the extension of the sugar chain in the process of peptidoglycan synthesis, thereby inhibiting bacterial growth.

2. A biopeptide antibacterial mouthwash, characterized in that: It also contains small molecule peptides that interact with the substrate binding site of the ATP-binding cassette transporter on the cell membrane of oral bacteria, preventing them from taking in nutrients and excreting harmful substances, thereby inhibiting bacterial proliferation.

3. A biopeptide antibacterial mouthwash, characterized in that: It also contains small molecule peptides that bind to proton motive force-related proteins on the cell membrane of oral bacteria, interfere with their normal conformation and function, destroy the energy conversion and material transport balance of the bacterial cell membrane, and thus inhibit bacterial growth.

4. A biopeptide antibacterial mouthwash, characterized in that: Also contains a slow-release organic compound: ethyl 2-(2-hydroxyphenoxy)acetate.

5. A biopeptide antibacterial mouthwash, characterized in that: Contains the following ingredients in weight percentage: SEQ NO.1: Leu-His-Pro-Gly-Tyr-Lys-Glu small molecule peptide: 0.15 parts to 0.35 parts; SEQ NO.2: Arg-Trp-Ser-Ile-Asp-Met-Cys small molecule peptide: 0.1-0.2 parts; SEQ NO.3: Phe-Ala-Val-Gln-Thr-Asn-Leu small molecule peptide: 0.1-0.2 parts; Moisturizer: 5-10 parts of glycerin and 0.1-0.3 parts of sodium hyaluronate; Surfactant: 1-3 parts of cocamidopropyl betaine; Flavoring: 0.1-0.3 parts of menthol and 2-5 parts of xylitol; Preservative: potassium sorbate 0.05-0.15 parts; Buffer: 0.2-0.5 parts of disodium hydrogen phosphate and 0.1-0.3 parts of citric acid; Slow-release organic compound: ethyl 2-(2-hydroxyphenoxy)acetate, 1 to 3 parts.

6. The biopeptide antibacterial mouthwash according to claim 1, wherein the SEQ NO.1: Leu-His-Pro-Gly-Tyr-Lys-Gl, SEQ NO.2: Arg-Trp-Ser-Ile-Asp-Met-Cys, SEQ NO.3 SEQ NO.3: Phe-Ala-Val-Gln-Thr-Asn-Leu.

7. The biopeptide antibacterial mouthwash according to claim 1, characterized in that: SEQ NO.1, SEQ NO.2 and SEQ NO.

3. Small molecule peptides were combined in a ratio of 3:2:2, with a total peptide concentration of 0.3 mg / mL.

8. A method for preparing the biopeptide antibacterial mouthwash according to any one of claims 1 to 8, characterized in that: The following steps are involved: S1 uses chemical synthesis to prepare SEQ NO.1, SEQ NO.2 and SEQ NO.3 small molecule peptides. During the synthesis process, a suitable solid phase synthetic resin is selected, and the corresponding amino acids are modified with protective groups, and then the amino acids are coupled to the resin in sequence. The reaction temperature is controlled at 20-30°C, the pH value is 6-8, and the coupling time of each amino acid is 1-2 hours. After the peptide chain is synthesized, the protective group is removed and cleaved, and then purified and identified by high performance liquid chromatography, so that the purity of the peptide reaches more than 95 parts; S2: mixing the purified SEQ NO.1, SEQ NO.2 and SEQ NO.3 small molecule peptides with a moisturizer, a surfactant, a flavoring agent, a preservative and a buffer in an appropriate amount of deionized water according to the ratio of claim 1, first dissolving the small molecule peptides in a small amount of deionized water, then adding other ingredients in sequence, and slowly stirring at a speed of 200-400 rpm in a stirrer for 30-60 minutes until uniform; S3 finally filters the mixed solution with a 0.22 μm microporous filter membrane to remove impurities, obtains the finished product of the bio-peptide antibacterial mouthwash, and performs aseptic packaging.

9. Use of the biopeptide antibacterial mouthwash according to any one of claims 1 to 7 in oral care, for preventing and improving oral diseases caused by Streptococcus mutans, Porphyromonas gingivalis and Fusobacterium nucleatum, including caries, gingivitis and halitosis.