Application of polypeptide in preparation of product for treating and / or preventing gingivitis and / or periodontitis

Abstract

The invention provides application of polypeptide in preparation of a product for treating and / or preventing gingivitis and / or periodontitis. The polypeptide with the amino acid sequence shown as SEQ ID NO: 1 is adopted for treating a gingival epithelial cell inflammation model, it is found that the polypeptide can effectively reduce the expression level of inflammatory factors IL-8, IL-1beta and TNF-alpha, and it is indicated that the polypeptide can effectively treat and / or prevent gingivitis and / or periodontitis and is suitable for preparing products for treating and / or preventing gingivitis and / or periodontitis.

Description

Technical Field

[0001] The present invention belongs to the technical field of polypeptides, and more specifically, relates to the use of polypeptides in preparing products for treating and / or preventing gingivitis and / or periodontitis. Background Art

[0002] Gingivitis is an inflammation of the gum tissue caused mainly by dental plaque. The bacteria in dental plaque can cause symptoms such as redness, swelling, bleeding, and pain in the gums. If not treated in time, gingivitis may develop into periodontitis, causing serious damage to the periodontal tissues (gingiva, periodontal ligament, alveolar bone, and cementum). Periodontitis is a chronic inflammation of the periodontal tissue caused mainly by dental plaque. The bacteria in dental plaque form dental calculus on the periodontal tissue, which stimulates and causes inflammation, leading to problems such as periodontal pocket formation, alveolar bone absorption, and loose teeth. Periodontitis not only affects the patient's chewing function, but may also cause systemic diseases such as cardiovascular disease and diabetes. It can be seen that gingivitis and periodontitis are both common inflammatory diseases of the oral cavity, which have important effects on oral health and overall health.

[0003] At present, the drugs commonly used to treat gingivitis and periodontitis include: cephalosporin antibiotics (such as cefradine, cephalexin, cefaclor, etc.), nitroimidazole antibiotics (such as metronidazole, tinidazole, ornidazole, etc.), penicillin antibiotics (such as penicillin V potassium tablets, amoxicillin capsules, etc.), etc. These antibiotics have good bactericidal effects, reduce inflammatory symptoms, and promote the recovery of gum health. However, long-term use of antibiotics may lead to adverse reactions and drug resistance.

[0004] Polypeptides are a type of compound formed by linking multiple α-amino acids together through a peptide chain. Due to their obvious advantages such as multiple biological activities and low risk of adverse reactions and drug resistance, they have gradually received widespread attention in the pharmaceutical field. However, there are still few polypeptides that can treat gingivitis or periodontitis. Summary of the invention

[0005] The present invention aims to provide the use of a polypeptide in the preparation of a product for treating and / or preventing gingivitis and / or periodontitis in view of the deficiencies of the prior art. The polypeptide can effectively reduce the expression levels of inflammatory factors IL-8, IL-1β, and TNF-α in a gingival epithelial cell inflammation model, thereby effectively treating and / or preventing gingivitis and / or periodontitis, providing a new raw material option for products for treating and / or preventing gingivitis and / or periodontitis, and also enriching the polypeptide resource library for treating and / or preventing gingivitis and / or periodontitis.

[0006] The first object of the present invention is to provide a use of a polypeptide in preparing a product for treating and / or preventing gingivitis and / or periodontitis.

[0007] The second objective of the present invention is to provide the use of polypeptide-related biomaterials in the preparation of products for treating and / or preventing gingivitis and / or periodontitis.

[0008] The third object of the present invention is to provide the use of active peptides in the preparation of products for treating and / or preventing gingivitis and / or periodontitis.

[0009] The fourth object of the present invention is to provide the use of biomaterials related to active peptides in the preparation of products for treating and / or preventing gingivitis and / or periodontitis.

[0010] The above-mentioned purpose of the present invention is achieved through the following technical solutions:

[0011] The present invention uses a polypeptide with an amino acid sequence as shown in SEQ ID NO: 1 to treat a gingival epithelial cell inflammation model, and finds that it can effectively reduce the expression levels of inflammatory factors IL-8, IL-1β, and TNF-α, indicating that the polypeptide of the present invention can effectively treat and / or prevent gingivitis and / or periodontitis, and is suitable for preparing products for treating and / or preventing gingivitis and / or periodontitis. Therefore, the present invention provides the following applications:

[0012] (1) Use of polypeptides in the preparation of products for treating and / or preventing gingivitis and / or periodontitis;

[0013] (2) Use of polypeptide-related biomaterials in the preparation of products for treating and / or preventing gingivitis and / or periodontitis;

[0014] (3) Use of active peptides in the preparation of products for the treatment and / or prevention of gingivitis and / or periodontitis;

[0015] (4) Application of biomaterials related to active peptides in the preparation of products for the treatment and / or prevention of gingivitis and / or periodontitis;

[0016] The amino acid sequence of the polypeptide is shown in SEQ ID NO: 1; the active peptide has more than 85% homology with the polypeptide with the amino acid sequence shown in SEQ ID NO: 1.

[0017] Optionally, the related biological material of the polypeptide is a nucleic acid molecule capable of expressing the polypeptide.

[0018] Optionally, the related biological material of the polypeptide is a recombinant cell, a recombinant bacterium, a recombinant plasmid or an expression cassette containing a nucleic acid molecule capable of expressing the polypeptide.

[0019] Optionally, the biological material related to the active peptide is a nucleic acid molecule capable of expressing the active peptide.

[0020] Optionally, the biological material related to the active peptide is a recombinant cell, a recombinant bacterium, a recombinant plasmid or an expression cassette containing a nucleic acid molecule capable of expressing the active peptide.

[0021] Optionally, the treatment and / or prevention of gingivitis and / or periodontitis is to reduce the expression level of inflammatory factors.

[0022] Furthermore, the inflammatory factor is one or more of IL-8, IL-1β, and TNF-α.

[0023] Optionally, the gingivitis is gingivitis caused by lipopolysaccharide stimulation.

[0024] Optionally, the periodontitis is periodontitis caused by lipopolysaccharide stimulation.

[0025] Optionally, the product is a medicine or an oral product.

[0026] Further, the preparation of the drug is one of oral liquid, spirit, tincture, aerosol, powder spray, injection, sterile powder for injection, and suppository. The types of the above preparations can be understood according to the relevant definitions in Pharmaceutics (Sixth Edition, People's Medical Publishing House, Cuifude), and the preparation of the above preparations can be prepared according to the methods of the relevant preparations in Pharmaceutics (Sixth Edition, People's Medical Publishing House, Cuifude).

[0027] Furthermore, the medicine also contains pharmaceutically acceptable excipients.

[0028] Furthermore, the auxiliary materials are solid lubricants (such as stearic acid and / or magnesium stearate, etc., used to reduce friction and ensure the success of drug manufacturing), vegetable oils (such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil and / or cocoa butter, etc., used as solvents or carriers to help drug ingredients dissolve or disperse better), polyols (such as propylene glycol, glycerol, sorbitol, mannitol and / or polyethylene glycol, etc., used to improve the stability of drugs, prevent drugs from losing water or deteriorating during storage, and also used to improve the taste and solubility of drugs), alginic acid (to maintain the physical stability of drugs, prevent To prevent drug particles from settling or agglomerating), emulsifiers (such as Tween and / or polyoxyethylene castor oil, etc., used to reduce interfacial tension, promote emulsification of the dispersed phase and maintain the stability of the emulsion), wetting agents (such as sodium lauryl sulfate, etc., used to help drug ingredients disperse and dissolve better), colorants (used to improve the appearance color of the drug, making it easier to identify and distinguish), flavoring agents (used to improve the taste of the drug and improve patient compliance with medication), tableting (favorable for pressing loose granular substances into solid tablets, which are convenient for patients to take and carry, and are also beneficial for improving the stability and bioavailability of drugs). The pharmaceutical composition may be selected from the group consisting of: a stabilizer (for increasing the physical stability of the pharmaceutical composition and preventing it from deteriorating or degrading during storage or transportation); an antioxidant (for preventing the pharmaceutical composition from deteriorating due to oxidation during storage or use and extending the shelf life of the pharmaceutical composition); an isotonic saline solution (for adjusting the osmotic pressure of the pharmaceutical composition to meet the physiological requirements of the human body, thereby reducing irritation and adverse reactions to the human body); a phosphate buffer solution (for adjusting the pH value of the pharmaceutical composition to meet the physiological requirements of the human body, thereby reducing irritation and adverse reactions to the human body); sugars (such as lactose, glucose and / or sucrose, etc., for helping the pharmaceutical composition to be better dispersed and dissolved, while providing the necessary energy support); starch (such as corn starch and / or potato starch, etc., for helping the tablet to disintegrate quickly after taking the tablet and release the pharmaceutical composition, thereby improving the bioavailability of the pharmaceutical composition); cellulose and its derivatives (such as sodium carboxymethyl cellulose, ethyl cellulose and / or methyl cellulose, etc., for improving the compressibility and formability of the pharmaceutical composition and for controlling the release rate of the pharmaceutical composition); an adhesive (such as gelatin, etc., for helping the pharmaceutical composition to be better combined together); and a lubricant (such as talc, etc., for reducing the friction of the pharmaceutical composition). The type of excipient can be selected according to the need to improve the stability, activity and / or bioavailability of the drug.

[0029] Furthermore, the oral product is one or more of oral lozenges, mouthwash effervescent tablets, oral care liquid, toothpaste or mouthwash.

[0030] Furthermore, the oral product also contains an auxiliary material acceptable to oral products.

[0031] Furthermore, the auxiliary materials are solvents (such as glycerol, propylene glycol and / or ethanol, etc., used to dissolve raw materials for easy use and storage), flavors (such as mint flavor, etc., used to increase the aroma and taste of oral products and enhance the sensory experience of users), preservatives (such as potassium sorbate, sodium benzoate, methylparaben and / or propylparaben, etc., used to prevent the growth of microorganisms in oral products and extend the shelf life of oral products), antibacterial agents (such as cetylpyridinium chloride, chlorhexidine gluconate and / or benzalkonium chloride, etc., used to inhibit or kill harmful microorganisms in the oral cavity, which helps maintain oral health), pH regulators (such as disodium hydrogen phosphate, sodium dihydrogen phosphate and / or sodium citrate, etc., used to adjust the pH of oral products, To make it meet the physiological needs of the oral environment and reduce the stimulation to the oral tissues), thickeners (such as poloxamer 407, etc., used to increase the viscosity of oral products, improve the use experience, and also used to improve the stability of oral products, not easy to stratify or precipitate), sweeteners (such as sucralose, sodium saccharin, sorbitol, xylitol and / or maltitol, etc., used to increase the sweetness of oral products and enhance the sensory experience of users), cooling agents (such as menthol and its derivatives, etc., used to increase the cooling feeling of oral products and enhance the sensory experience of users), surfactants (such as sodium lauryl sulfate and / or sodium lauroyl sarcosine, etc., used to reduce the surface tension of oral products and enhance the cleaning effect) one or more. The type of the auxiliary material can be selected according to the needs of improving the stability, safety, effectiveness or improving the use experience of oral products.

[0032] The present invention has the following beneficial effects:

[0033] The present invention uses a polypeptide with an amino acid sequence as shown in SEQ ID NO: 1 to treat a gingival epithelial cell inflammation model, and finds that it can effectively reduce the expression levels of inflammatory factors IL-8, IL-1β, and TNF-α, indicating that the polypeptide of the present invention can effectively treat and / or prevent gingivitis and / or periodontitis, and is suitable for preparing products for treating and / or preventing gingivitis and / or periodontitis. Therefore, the present invention provides the use of a polypeptide with an amino acid sequence as shown in SEQ ID NO: 1 in preparing products for treating and / or preventing gingivitis and / or periodontitis. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 This is the gene expression of inflammatory factors (IL-8, IL-1β and TNF-α) in Example 2. DETAILED DESCRIPTION

[0035] The present invention is further described below in conjunction with the accompanying drawings and specific embodiments of the specification, but the embodiments do not limit the present invention in any form. It should be understood that these embodiments and examples are only used to illustrate the present invention and are not used to limit the scope of the present invention. The purpose of providing these embodiments and examples is to make the understanding of the disclosure of the present invention more thorough and comprehensive. It should also be understood that the present invention can be implemented in many different forms and is not limited to the embodiments and examples described herein. Those skilled in the art can make various changes or modifications without violating the connotation of the present invention, and the equivalent forms obtained also fall within the protection scope of the present invention. For example, the features described or described as part of an embodiment can be combined in another embodiment in a suitable manner to produce a new embodiment. In addition, in the description below, a large number of specific details are given in order to provide a more comprehensive understanding of the present invention. It should be understood that the present invention can be implemented without one or more of these details. Unless otherwise specified, the reagents, methods and equipment used in the embodiments of the present invention are conventional reagents, methods and equipment in the art. Unless otherwise specified, the reagents and materials used in the embodiments of the present invention are commercially available.

[0036] Example 1 Synthesis of polypeptide

[0037] The polypeptide was synthesized by the Sangon Bioengineering Company, and the amino acid sequence of the polypeptide is shown in SEQ ID NO:1.

[0038] Among them, SEQ ID NO:1: GLDWWQL.

[0039] Example 2 Polypeptides can effectively treat and / or prevent gingivitis and / or periodontitis

[0040] 1. Experimental grouping and treatment:

[0041] (1) Experimental group (LPS+12.5μM peptide, LPS+25μM peptide):

[0042] Human gingival epithelial cells (HGECs) were taken out of liquid nitrogen, thawed in a 37°C water bath, and inoculated into DMEM medium containing 10% (v / v) fetal bovine serum and 1% (v / v) penicillin / streptomycin. After centrifugation at 1000 rpm for 5 min, the cells were collected and resuspended in DMEM medium containing 10% (v / v) fetal bovine serum and 1% (v / v) penicillin / streptomycin. 5 Gingival epithelial cells were seeded in 6-well plates at a density of 10 cells / mL and incubated at 37°C and 5% CO 2After the cells grew to 70% under the conditions, Porphyromonas gingivalis lipopolysaccharide (LPS, to a final concentration of 1.0 μg / mL, and the 6-well plate was shaken to evenly distribute the Porphyromonas gingivalis lipopolysaccharide) and the polypeptide obtained in Example 1 (to a final concentration of 12.5 and 25 μM, respectively) were added to each well, and the culture was continued at 37°C and 5% CO 2 After culturing in an incubator for 24 h, RNA of human gingival epithelial cells was extracted and the gene expression of inflammatory factors (IL-8, IL-1β and TNF-α) was analyzed using reverse transcription-polymerase chain reaction (RT-PCR) technology.

[0043] (2) Blank group (CON):

[0044] Human gingival epithelial cells (HGECs) were taken out of liquid nitrogen, thawed in a 37°C water bath, and inoculated into DMEM medium containing 10% (v / v) fetal bovine serum and 1% (v / v) penicillin / streptomycin. After centrifugation at 1000 rpm for 5 min, the cells were collected and resuspended in DMEM medium containing 10% (v / v) fetal bovine serum and 1% (v / v) penicillin / streptomycin. 5 Gingival epithelial cells were seeded in 6-well plates at a density of 10 cells / mL and incubated at 37°C and 5% CO 2 After the cells grew to 70% growth under the conditions, they were cultured at 37°C and 5% CO 2 After culturing in an incubator for 24 h, RNA of human gingival epithelial cells was extracted and the gene expression of inflammatory factors (IL-8, IL-1β and TNF-α) was analyzed using reverse transcription-polymerase chain reaction (RT-PCR) technology.

[0045] (3) Control group (LPS):

[0046] Human gingival epithelial cells (HGECs) were taken out of liquid nitrogen, thawed in a 37°C water bath, and inoculated into DMEM medium containing 10% (v / v) fetal bovine serum and 1% (v / v) penicillin / streptomycin. After centrifugation at 1000 rpm for 5 min, the cells were collected and resuspended in DMEM medium containing 10% (v / v) fetal bovine serum and 1% (v / v) penicillin / streptomycin. 5 Gingival epithelial cells were seeded in 6-well plates at a density of 10 cells / mL and incubated at 37°C and 5% CO 2 After the cells grew to 70% under the conditions, Porphyromonas gingivalis lipopolysaccharide (LPS) was added to each well to make the final concentration of 1.0 μg / mL, and the 6-well plate was shaken to evenly distribute the Porphyromonas gingivalis lipopolysaccharide. The culture was continued at 37°C and 5% CO. 2After culturing in an incubator for 24 h, RNA of human gingival epithelial cells was extracted and the gene expression of inflammatory factors (IL-8, IL-1β and TNF-α) was analyzed using reverse transcription-polymerase chain reaction (RT-PCR) technology.

[0047] 2. Experimental results:

[0048] Gene expression of inflammatory factors Figure 1 As shown. Figure 1 It can be seen that compared with the blank group, the expression levels of inflammatory factors IL-8, IL-1β, and TNF-α in human gingival epithelial cells in the control group under the stimulation of LPS were significantly increased, while the experimental group treated with the polypeptide obtained in Example 1 can significantly reduce the expression levels of inflammatory factors IL-8, IL-1β, and TNF-α, indicating that the polypeptide of the present invention can effectively inhibit the inflammation of gingival epithelial cells caused by LPS stimulation, thereby effectively treating and / or preventing gingivitis and / or periodontitis.

[0049] In summary, the present invention uses a polypeptide with an amino acid sequence as shown in SEQ ID NO: 1 to treat a gingival epithelial cell inflammation model, and it is found that it can effectively reduce the expression levels of inflammatory factors IL-8, IL-1β, and TNF-α, indicating that the polypeptide of the present invention can effectively treat and / or prevent gingivitis and / or periodontitis, and provides a new raw material selection for products for treating and / or preventing gingivitis and / or periodontitis, and also enriches the polypeptide resource library that has the effect of treating and / or preventing gingivitis and / or periodontitis.

[0050] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. Use of a polypeptide in the preparation of a product for treating and / or preventing gingivitis and / or periodontitis, characterized in that: The amino acid sequence of the polypeptide is shown in SEQ ID NO:

1.

2. Use of polypeptide-related biomaterials in the preparation of products for treating and / or preventing gingivitis and / or periodontitis, characterized in that: The amino acid sequence of the polypeptide is shown in SEQ ID NO:

1.

3. The application according to claim 2, characterized in that: The relevant biological material of the polypeptide is a nucleic acid molecule capable of expressing the polypeptide.

4. The application according to claim 2, characterized in that: The related biological material of the polypeptide is a recombinant cell, a recombinant bacterium, a recombinant plasmid or an expression box containing a nucleic acid molecule capable of expressing the polypeptide.

5. Use of an active peptide in the preparation of a product for treating and / or preventing gingivitis and / or periodontitis, characterized in that: The active peptide has more than 85% homology with the polypeptide with an amino acid sequence as shown in SEQ ID NO:

1.

6. Use of biomaterials related to active peptides in the preparation of products for treating and / or preventing gingivitis and / or periodontitis, characterized in that: The active peptide has more than 85% homology with the polypeptide with an amino acid sequence as shown in SEQ ID NO:

1.

7. The use according to claim 6, characterized in that: The biological material related to the active peptide is a nucleic acid molecule capable of expressing the active peptide.

8. The use according to claim 6, characterized in that: The biological material related to the active peptide is a recombinant cell, a recombinant bacterium, a recombinant plasmid or an expression box containing a nucleic acid molecule capable of expressing the active peptide.

9. The use according to any one of claims 1 to 8, characterized in that: The treatment and / or prevention of gingivitis and / or periodontitis is to reduce the expression level of inflammatory factors.

10. The use according to claim 9, characterized in that: The inflammatory factor is one or more of IL-8, IL-1β, and TNF-α.

Images