Artemisinin compound periodontitis inhibition and mucous membrane repair composition

Abstract

The invention discloses an artemisinin compound type composition for inhibiting periodontitis and repairing mucous membranes. The invention relates to the technical field of periodontitis repair compositions. The composition comprises an antibacterial component and formylated hyaluronic acid. According to the artemisinin compound type periodontitis inhibition and mucous membrane repair composition, the antibacterial component can be combined with protein of bacteria to denature the protein of the bacteria, so that the growth and reproduction of the bacteria are inhibited, in treatment of periodontitis, the number of harmful bacteria in the oral cavity can be reduced, meanwhile, the inhibition effect on the bacteria causing periodontitis can be enhanced, and the inflammatory response can be relieved; the sulfonated chitosan, the L-ascorbic acid and other components can promote the tissue repair function, can be adsorbed on the mucous membrane surface in oral mucous membrane repair, provide a support for cell attachment and growth, facilitate migration and proliferation of damaged mucous membrane cells, promote synthesis of collagen, facilitate repair of damaged periodontal tissues and mucous membranes and improve the oral mucous membrane repair effect. The wound healing is accelerated.

Description

Technical Field

[0001] The invention relates to the technical field of periodontitis repair compositions, and in particular to an artemisinin-compounded periodontitis inhibition and mucosal repair composition. Background Art

[0002] Artemisinin is a sesquiterpene lactone compound containing a peroxy bridge structure extracted from the plant Artemisia annua. It has antimicrobial activity. It can destroy the cell structure and function of periodontal pathogens, help reduce the number of periodontal pathogens in the oral cavity, reduce the inflammatory response caused by these bacteria, and help control the growth and reproduction of common periodontal pathogens such as Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans, thereby alleviating symptoms such as gingival redness, swelling and bleeding, and helping to improve periodontal health.

[0003] The Chinese patent with publication number CN106214505A discloses a new application of artemisinin components or their compositions, namely, a new application in the field of oral care for mammals. It eliminates or alleviates the adverse conditions of the mammalian oral cavity by providing a series of benefits such as correction of abnormal keratinization, anti-inflammatory, analgesic, hemostatic, healing, antibacterial, and antifungal, so as to enhance the health and beauty of the oral cavity. The benefits are demonstrated by significantly preventing or alleviating dental plaque, gingivitis, periodontitis, oral keratosis, tooth sensitivity, oral ulcers, caries, and proliferation of oral pathogens in mammals; the benefits are achieved by administering artemisinin components or their compositions to mammals or oral care compositions containing artemisinin components or their compositions, such as health products, daily chemical products, health products, or medicines. The invention expands the use of artemisinin components or their compositions in the field of oral care, so that they can play a greater role in the health of the general public. However, the composition prepared by the above technology has limited effect on the repair of the mucosa when used for periodontitis; therefore, the present invention proposes an artemisinin-compounded periodontitis inhibition and mucosal repair composition to solve the above-mentioned problems. Summary of the invention

[0004] 1. Technical issues to be resolved In view of the deficiencies in the prior art, the present invention provides an artemisinin-compounded periodontitis inhibition and mucosal repair composition, which solves the problems mentioned in the above background technology.

[0005] (II) Technical solution To achieve the above objectives, the present invention is implemented through the following technical solutions: an artemisinin-compounded periodontitis inhibition and mucosal repair composition, comprising an antibacterial component and aldehyde-modified hyaluronic acid; Wherein, the antibacterial component is compounded by copper sulfate, artemisinin and sulfonated chitosan; The preparation method of the artemisinin-based periodontitis inhibition and mucosal repair composition comprises the following steps: dissolving an antibacterial component and aldehyde-modified hyaluronic acid in 1-3 mL of a PBS buffer solution at a mass ratio of (3-5):7, then pouring the solution into a mold at room temperature, stirring evenly for 10-15 seconds, and standing the mixture for 15-30 minutes to obtain the artemisinin-based periodontitis inhibition and mucosal repair composition.

[0006] Preferably, the preparation method of the antibacterial component is: (1) Add copper sulfate to deionized water and stir until it is completely dissolved to prepare solution A; (2) Pour artemisinin into a 1%-5% acetic acid solution and stir until it is completely dissolved to prepare solution B; (3) Pour sulfonated chitosan into deionized water, stir for 7-10 min at room temperature, add L-ascorbic acid, and continue stirring for 30-40 min to obtain solution C; (4) Heat solution C in a water bath, and add solution A and solution B thereto under stirring. Continue stirring and reacting for 30-40 minutes to obtain the antibacterial component.

[0007] Preferably, in said (4), the temperature is raised to 80-85°C.

[0008] The mass ratio of the copper sulfate, artemisinin, sulfonated chitosan and L-ascorbic acid is (0.2-0.3): (0.25-0.4): (0.1-0.3): (0.12-0.18).

[0009] Preferably, in (3), the preparation method of sulfonated chitosan is: (3.1) Pour chitosan into 1%-5% acetic acid and stir to react for 45-60 minutes; (3.2) Slowly add 1,3-propane sultone, heat the water bath to 60-70°C, and stir for 5-6 hours; (3.3) Cool to room temperature, then slowly add dropwise to the acetone solution. After the reaction is completed, wash the reaction product with anhydrous methanol for 3-5 times, and then dissolve it in distilled water; (3.4) and finally placed in a vacuum freeze dryer for 42-48 hours to obtain sulfonated chitosan.

[0010] Preferably, in said (3.4), the cold treatment temperature is -22--25°C, and the cold treatment time is 20-24h.

[0011] Preferably, the mass ratio of chitosan to 1,3-propane sultone is (1-2.5): (1-2.5).

[0012] Preferably, the preparation method of the aldehyded hyaluronic acid is: S1. Heat deionized water, then pour in hyaluronic acid and stir until completely dissolved; S2. Add sodium periodate under light-proof conditions and stir to react for 1.5-2h; S3, slowly add ethylene glycol and continue stirring the reaction for 40-60 minutes; S4. Dialyze in deionized water for 3-4 days using a dialysis bag, and then freeze-dry to obtain aldehyde-modified hyaluronic acid.

[0013] Preferably, in S1, the temperature is raised to 55-65°C.

[0014] Preferably, in S4, the molecular weight cutoff of the dialysis bag is 8000-14000Da.

[0015] Preferably, the mass ratio of hyaluronic acid to sodium periodate is (1.2-2.5):(0.5-0.8).

[0016] (III) Beneficial effects The present invention provides an artemisinin-based periodontitis inhibition and mucosal repair composition. Compared with the prior art, it has the following beneficial effects: (1) The artemisinin-based periodontitis inhibition and mucosal repair composition has antibacterial ingredients that can bind to bacterial proteins to denature them, thereby inhibiting bacterial growth and reproduction. In the treatment of periodontitis, it can help reduce the number of harmful bacteria in the oral cavity, while enhancing the inhibitory effect on bacteria that cause periodontitis and reducing inflammatory responses. In addition, ingredients such as sulfonated chitosan and L-ascorbic acid can promote tissue repair. In oral mucosal repair, they can be adsorbed on the mucosal surface to provide a scaffold for cell attachment and growth, which is beneficial to the migration and proliferation of damaged mucosal cells. They can also promote the synthesis of collagen, which helps repair damaged periodontal tissues and mucosa and accelerate wound healing.

[0017] (2) The artemisinin-based periodontitis inhibition and mucosal repair composition, the aldehyde-modified hyaluronic acid and other components work synergistically, which is beneficial to the adhesion and proliferation of cells, can provide a good environment for mucosal repair, and promote the growth and repair of oral mucosal cells; it can also maintain the moisture of the oral mucosa, which is beneficial to the self-repair of damaged mucosa. At the same time, the gel-like substance it forms can form a protective film on the mucosal surface to prevent further invasion by harmful substances such as bacteria, and has a good synergistic effect on the inhibition of periodontitis and mucosal repair. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This is a comparison chart of the antibacterial performance test provided by the present invention. DETAILED DESCRIPTION

[0019] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0020] Example 1 The antibacterial component and the aldehyde-modified hyaluronic acid were dissolved in 1 mL of PBS buffer solution at a mass ratio of 3:7, and then poured into a mold at room temperature, stirred evenly for 10 seconds, and allowed to stand for 15 minutes to obtain an artemisinin-based periodontitis inhibition and mucosal repair composition; The preparation method of the antibacterial component is as follows: pour 0.2g of copper sulfate into 5mL of deionized water and stir until it is completely dissolved to obtain solution A; pour 0.25g of artemisinin into 3.5mL of acetic acid solution with a mass fraction of 1% and stir until it is completely dissolved to obtain solution B; pour 0.1g of sulfonated chitosan into 42mL of deionized water, stir for 7min at room temperature, add 0.12g of L-ascorbic acid, and continue stirring for 30min to obtain solution C; heat solution C to 80°C in a water bath, add solution A and solution B thereto under stirring, and continue stirring and reacting for 30min to obtain the antibacterial component; The preparation method of sulfonated chitosan is as follows: pour 1g of chitosan into 80mL of acetic acid with a mass fraction of 4%, and stir to react for 45min; slowly add 2.5g of 1,3-propane sultone, heat to 60°C in a water bath, and stir for 5h; cool to room temperature, and then slowly drop into an acetone solution. After the reaction is completed, wash the reaction product with anhydrous methanol for 3 times, and then dissolve it in distilled water; place it under a temperature condition of -22°C for 20h, and finally place it in a vacuum freeze dryer for 42h to obtain sulfonated chitosan; The preparation method of aldehyded hyaluronic acid is as follows: heat 100 mL of deionized water to 55°C, then pour in 1.2 g of hyaluronic acid and stir until completely dissolved; add 0.5 g of sodium periodate under light-proof conditions and stir to react for 1.5 hours; slowly add 1 mL of ethylene glycol and continue to stir to react for 40 minutes; dialyze in deionized water for 3 days using a dialysis bag with a molecular weight cutoff of 8000 Da, and then freeze-dry to obtain aldehyded hyaluronic acid.

[0021] Example 2 The antibacterial component and the aldehyde-modified hyaluronic acid were dissolved in 2 mL of PBS buffer solution at a mass ratio of 4:7, and then poured into a mold at room temperature, stirred evenly for 12 seconds, and allowed to stand for 25 minutes to obtain an artemisinin-compounded periodontitis inhibition and mucosal repair composition; The preparation method of the antibacterial component is as follows: pour 0.25g of copper sulfate into 5mL of deionized water and stir until it is completely dissolved to obtain solution A; pour 0.3g of artemisinin into 3.5mL of acetic acid solution with a mass fraction of 3% and stir until it is completely dissolved to obtain solution B; pour 0.2g of sulfonated chitosan into 42mL of deionized water, stir for 8min at room temperature, add 0.15g of L-ascorbic acid, and continue stirring for 35min to obtain solution C; heat solution C to 82°C in a water bath, add solution A and solution B thereto under stirring, and continue stirring for 35min to obtain the antibacterial component; The preparation method of sulfonated chitosan is as follows: pour 1.5g of chitosan into 80mL of acetic acid with a mass fraction of 2%, and stir to react for 50min; slowly add 2.5g of 1,3-propane sultone, heat to 65°C in a water bath, and stir for 5.5h; cool to room temperature, and then slowly drop into an acetone solution. After the reaction is completed, wash the reaction product with anhydrous methanol for 4 times, and then dissolve it in distilled water; place it under a temperature condition of -23°C for 22h, and finally place it in a vacuum freeze dryer for 45h to obtain sulfonated chitosan; The preparation method of aldehyded hyaluronic acid is as follows: heat 100 mL of deionized water to 60°C, then pour in 2 g of hyaluronic acid and stir until completely dissolved; add 0.6 g of sodium periodate under light-proof conditions and stir to react for 1.8 hours; slowly add 1.3 mL of ethylene glycol and continue to stir to react for 50 minutes; dialyze in deionized water for 4 days using a dialysis bag with a molecular weight cutoff of 10,000 Da, and then freeze-dry to obtain aldehyded hyaluronic acid.

[0022] Example 3 The antibacterial component and the aldehyde-modified hyaluronic acid were dissolved in 3 mL of PBS buffer solution at a mass ratio of 5:7, and then poured into a mold at room temperature, stirred evenly for 15 seconds, and allowed to stand for 30 minutes to obtain an artemisinin-based periodontitis inhibition and mucosal repair composition; The preparation method of the antibacterial component is as follows: 0.3 g of copper sulfate is poured into 5 mL of deionized water and stirred until it is completely dissolved to obtain solution A; 0.4 g of artemisinin is poured into 3.5 mL of acetic acid solution with a mass fraction of 5% and stirred until it is completely dissolved to obtain solution B; 0.3 g of sulfonated chitosan is poured into 42 mL of deionized water, stirred for 10 minutes at room temperature, and then 0.18 g of L-ascorbic acid is added, and stirring is continued for 40 minutes to obtain solution C; solution C is heated to 85° C. in a water bath, and solution A and solution B are added thereto under stirring conditions, and stirring is continued for 40 minutes to obtain the antibacterial component; The preparation method of sulfonated chitosan is as follows: pour 2.5g of chitosan into 80mL of acetic acid with a mass fraction of 5%, and stir to react for 60min; slowly add 2.5g of 1,3-propane sultone, heat to 70°C in a water bath, and stir for 6h; cool to room temperature, and then slowly drop into an acetone solution. After the reaction is completed, wash the reaction product with anhydrous methanol for 5 times, and then dissolve it in distilled water; place it under a temperature condition of -25°C for 24h, and finally place it in a vacuum freeze dryer for 48h to obtain sulfonated chitosan; The preparation method of aldehyded hyaluronic acid is as follows: heat 100 mL of deionized water to 65°C, then pour in 2.5 g of hyaluronic acid and stir until completely dissolved; add 0.8 g of sodium periodate under light-proof conditions and stir to react for 2 hours; slowly add 1.6 mL of ethylene glycol and continue to stir to react for 60 minutes; dialyze in deionized water for 4 days using a dialysis bag with a molecular weight cutoff of 14,000 Da, and then freeze-dry to obtain aldehyded hyaluronic acid.

[0023] Comparative Example 1 Compared with Example 1, the difference is that artemisinin is not added; the rest remains unchanged.

[0024] Comparative Example 2 Compared with Example 1, the difference is that the aldehyde-modified hyaluronic acid is replaced by hyaluronic acid; the rest remains unchanged.

[0025] The above-mentioned weight and milliliters are for single-dose mixing. If a large dose is to be mixed, the amount added can be increased accordingly.

[0026] After anesthesia, the first molar on the right side of the experimental rats was used as the model tooth, and the periodontitis model was established using the double-line method supplemented with a high-sugar diet. The specific operation was as follows: 0# medical suture was buried in the gingival sulcus, and then the tooth neck was ligated with a 0.20mm orthodontic ligature. During this period, all rats were fed with a high-sugar feed, and regular inspections were conducted. The loose and detached ligatures were re-ligated in time. After 8 weeks, periodontitis was formed in the rats, and the high-sugar diet was stopped; the rats were anesthetized, the ligatures and sutures were removed, and 1% of the composition was used. The administration method was injection into the periodontal pocket until the drug overflowed from the periodontal pocket, and the frequency was once a week; the rats were anesthetized after the 0th, 2nd, and 4th weeks of administration, and the gingival state was observed and the depth of the periodontal pocket was probed; the results are shown in Table 1.

[0027] 0 Weeks 2 weeks 4 weeks Example 1 1.55±0.04 0.90±0.14 0.13±0.01 Example 2 1.59±0.04 0.93±0.13 0.15±0.07 Example 3 1.61±0.05 0.95±0.08 0.08±0.04 Comparative Example 1 1.67±0.08 1.39±0.13 1.31±0.09 Comparative Example 2 1.66±0.10 1.37±0.09 1.22±0.18 As shown in Table 1, there was little difference in the gum status of each group at week 0. After 2 weeks of medication, the gum redness and swelling of Examples 1-3 were alleviated, and the probing depth decreased. After 4 weeks, compared with Comparative Examples 1-2, the gum inflammation of Examples 1-3 was significantly subsided, and the probing depth was significantly reduced.

[0028] Antibacterial performance test: dilute the bacterial solution with LB medium to a concentration of 1x106 CFU / mL, add equal volumes of bacterial solution and sample into a 96-well plate, the total addition volume is 200 μL / well, and the final bacterial solution concentration is 5x10 5 CFU / mL, place the 96-well plate in a 37°C incubator and shake at 90 rpm for 24 h, measure the optical density at 600 nm with an ELISA reader, and calculate the inhibition rate, see Figure 1 It can be seen that the antibacterial rates of Examples 1, 2, and 3 against Staphylococcus aureus, Escherichia coli, and Porphyromonas are all above 97%, while the antibacterial rates of Comparative Examples 1 and 2 are only 85%-88%. Therefore, the antibacterial rate of the composition prepared by the preparation method of the present invention is significantly better than that of the comparative example.

[0029] Meanwhile, the contents not described in detail in this specification belong to the prior art known to those skilled in the art.

[0030] It should be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device.

[0031] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. An artemisinin-based periodontitis inhibition and mucosal repair composition, characterized in that: Includes antimicrobial ingredients, aldehyde-formylated hyaluronic acid; Wherein, the antibacterial component is compounded by copper sulfate, artemisinin and sulfonated chitosan; The preparation method of the artemisinin-based periodontitis inhibition and mucosal repair composition comprises the following steps: dissolving an antibacterial component and aldehyde-modified hyaluronic acid in 1-3 mL of a PBS buffer solution at a mass ratio of (3-5):7, then pouring the solution into a mold at room temperature, stirring evenly for 10-15 seconds, and standing the mixture for 15-30 minutes to obtain the artemisinin-based periodontitis inhibition and mucosal repair composition.

2. The artemisinin-based periodontitis inhibition and mucosal repair composition according to claim 1, characterized in that: The preparation method of the antibacterial component is: (1) Add copper sulfate to deionized water and stir until it is completely dissolved to prepare solution A; (2) Pour artemisinin into a 1%-5% acetic acid solution and stir until it is completely dissolved to prepare solution B; (3) Pour sulfonated chitosan into deionized water, stir for 7-10 min at room temperature, add L-ascorbic acid, and continue stirring for 30-40 min to obtain solution C; (4) Heat solution C in a water bath, and add solution A and solution B thereto under stirring. Continue stirring and reacting for 30-40 minutes to obtain the antibacterial component.

3. The artemisinin-based periodontitis inhibition and mucosal repair composition according to claim 2, characterized in that: In the step (4), the temperature is raised to 80-85°C.

4. The artemisinin-based periodontitis inhibition and mucosal repair composition according to claim 2, characterized in that: In (3), the preparation method of sulfonated chitosan is: (3.1) Pour chitosan into 1%-5% acetic acid and stir to react for 45-60 minutes; (3.2) Slowly add 1,3-propane sultone, heat the water bath to 60-70°C, and stir for 5-6 hours; (3.3) Cool to room temperature, then slowly add dropwise to the acetone solution. After the reaction is completed, wash the reaction product with anhydrous methanol for 3-5 times, and then dissolve it in distilled water; (3.4) and finally placed in a vacuum freeze dryer for 42-48 hours to obtain sulfonated chitosan.

5. The artemisinin-based periodontitis inhibition and mucosal repair composition according to claim 4, characterized in that: In the above (3.4), the cold treatment temperature is -22-25°C, and the cold treatment time is 20-24h.

6. The artemisinin-based periodontitis inhibition and mucosal repair composition according to claim 1, characterized in that: The preparation method of the aldehyde-modified hyaluronic acid is as follows: S1. Heat deionized water, then pour in hyaluronic acid and stir until completely dissolved; S2. Add sodium periodate under light-proof conditions and stir to react for 1.5-2h; S3, slowly add ethylene glycol and continue stirring the reaction for 40-60 minutes; S4. Dialyze in deionized water for 3-4 days using a dialysis bag, and then freeze-dry to obtain aldehyde-modified hyaluronic acid.

7. The artemisinin-based periodontitis inhibition and mucosal repair composition according to claim 4, characterized in that: In S1, the temperature is raised to 55-65°C.

8. The artemisinin-based periodontitis inhibition and mucosal repair composition according to claim 4, characterized in that: In the S4, the molecular weight cut-off of the dialysis bag is 8000-14000Da.

Images