Artemisinin component compounded periodontal pathogenic bacterium inhibiting and balancing agent

Abstract

The invention discloses an artemisinin component compounded periodontal pathogenic bacterium inhibition balance agent, and relates to the technical field of dental bacterium inhibitors, and the main components of the periodontal pathogenic bacterium inhibition balance agent comprise modified porous corn starch, chitosan and artemisinin. According to the artemisinin component compounded periodontal pathogenic bacterium inhibition balance agent, the porous corn starch has a porous structure and a large specific surface area, and after the porous corn starch is modified by octenyl succinic anhydride, the structural characteristic is still kept, so that harmful substances such as bacteria and toxins in the oral cavity can be adsorbed, and the number of periodontal pathogenic bacteria in the oral cavity is reduced; meanwhile, the modified porous corn starch is used as a carrier of the antibacterial component, and the antibacterial component can be well loaded on the starch, so that the antibacterial component can be continuously and slowly released in the oral cavity environment, the inhibition effect on periodontal pathogenic bacteria is maintained for a long time, and all the antibacterial components are not completely released at a time.

Description

Technical Field

[0001] The invention relates to the technical field of dental bacteria inhibitors, and in particular to a periodontal pathogenic bacteria inhibitory balancing agent compounded with an artemisinin component. Background Art

[0002] Periodontal pathogens refer to bacteria that can cause inflammation of periodontal tissues. Common periodontal pathogens include Porphyromonas gingivalis, which can produce a variety of virulence factors, destroy the connection of periodontal tissues, cause gingival inflammation and alveolar bone absorption; Actinobacillus actinomycetemcomitans is also a major periodontal pathogen, which is closely related to aggressive periodontitis, can inhibit the host's defense response, and can also invade the periodontal tissue cells to cause damage; in addition, Tannerella forsythiae is also relatively common, which can produce some enzymes to decompose proteins and other components in periodontal tissues; these bacteria interact with each other and jointly endanger periodontal health.

[0003] 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.

[0004] However, due to the fluidity of saliva, most drugs only stay in the oral cavity for a short time and cannot exert their efficacy for a long time, thus limiting the bactericidal effect. To this end, the present invention proposes a periodontal pathogen inhibition balancer compounded with artemisinin components to solve the above-mentioned problems. Summary of the invention

[0005] 1. Technical issues to be resolved In view of the deficiencies of the prior art, the present invention provides a periodontal pathogen inhibition balancing agent compounded with artemisinin components, which solves the problems mentioned in the above background technology.

[0006] (II) Technical solution To achieve the above objectives, the present invention is implemented by the following technical scheme: a periodontal pathogen inhibition balance agent compounded with artemisinin components, the main components of the periodontal pathogen inhibition balance agent include modified porous corn starch, chitosan, and artemisinin, and the preparation method thereof is: Ⅰ. Pour zinc acetate dihydrate into deionized water and stir until completely dissolved to prepare solution A; Ⅱ. Pour artemisinin into glacial acetic acid solution and stir until completely dissolved to obtain solution B; III. Pour chitosan into deionized water, stir at room temperature, then add L-ascorbic acid, continue stirring, and heat in a water bath to obtain solution C; Preferably, the stirring time at room temperature is 5-10 min, the stirring time is continued for 30-40 min, the temperature of the water bath is 70-80° C., and the heating time in the water bath is 10-20 min.

[0007] IV. Add solution A and solution B to solution C, continue heating in a water bath, add modified porous corn starch, stir evenly, and pour into a homogenizer for homogenization to obtain a periodontal pathogen inhibition balancer; Preferably, the water bath heating time is 6-8 hours and the homogenization time is 8-15 minutes.

[0008] Preferably, the mass ratio of zinc acetate dihydrate, artemisinin, chitosan, L-ascorbic acid and modified porous corn starch is: 0.1-0.16: 0.2-0.24g: 0.1-0.18g: 2.4-3.2g: 6.5-10g.

[0009] Preferably, the preparation method of the modified porous corn starch is: S1. Pour porous corn starch into distilled water, stir and mix evenly to prepare a starch dispersion with a mass fraction of 30%-35%; S2. Place the mixture in a constant temperature water bath and heat it to 50-60°C, stir for 20-30 minutes, then add 2-5% by mass NaOH solution to adjust the pH value of the starch dispersion to 8.5-9.0; S3, slowly dropwise add octenyl succinic anhydride diluted 3-4 times with anhydrous ethanol, and react in a water bath at 35-45°C for 3-4h; S4. After the reaction is completed, filter the mixture, wash the precipitate with 2-5% HCI solution until it is neutral, and then wash it with anhydrous ethanol for 3-5 times. Finally, dry it in a drying oven at 40-50°C for 18-24 hours, take it out, grind and sieve it to obtain modified porous corn starch.

[0010] Preferably, in S1, the mass ratio of porous corn starch to distilled water is 1:(1.8-2.3).

[0011] Preferably, in S3, the amount of octenyl succinic anhydride added is 3%-5% of the mass of the porous starch, and during the reaction, the pH value of the solution is controlled to be 8.2-8.6.

[0012] Preferably, the preparation method of the porous corn starch is: (1) Pour corn starch into the buffer solution and then place it in a water bath; (2) Add complex enzyme and place in a constant temperature shaker for reaction; (3) After the reaction is completed, add NaOH solution, filter, wash the precipitate with distilled water, dry it in a drying oven to constant weight, and finally grind and sieve to obtain porous corn starch.

[0013] Preferably, in (1), the buffer solution is selected from any one of disodium hydrogen phosphate and potassium dihydrogen phosphate, and the pH value is 5-6.

[0014] Preferably, in (2), the complex enzyme is composed of α-amylase and saccharifying enzyme, and the mass ratio thereof is 1.2:(0.7-1.1); the water bath temperature is 45-55°C, and the constant temperature heating is performed for 20-30 minutes.

[0015] Preferably, in (2), the temperature of the constant temperature shaker is 40-50°C, and the reaction is carried out at a rotation speed of 100-140 r / min for 8-10 hours; in (3), the drying temperature is 45-50°C.

[0016] (III) Beneficial effects The present invention provides a periodontal pathogenic bacteria inhibitory balance agent compounded with artemisinin components. Compared with the prior art, it has the following beneficial effects: (1) The artemisinin-based periodontal pathogen inhibitory balancer has a porous structure and a large specific surface area. After being modified with octenyl succinic anhydride, this structural feature is still retained, and it can adsorb harmful substances such as bacteria and toxins in the oral cavity, thereby reducing the number of periodontal pathogens in the oral cavity. At the same time, the modified porous corn starch is used as a carrier of antibacterial ingredients. The antibacterial ingredients can be well loaded on the starch, so that it can continuously and slowly release the antibacterial ingredients in the oral environment, maintaining the inhibitory effect on periodontal pathogens for a long time, rather than releasing all the antibacterial ingredients at once. Corn starch is a natural, biodegradable material. After modification, it still has good biocompatibility and is relatively safe to use in the oral cavity.

[0017] (2) The periodontal pathogen inhibitory balance agent compounded with artemisinin has good antibacterial and anti-inflammatory effects, and has inhibitory effects on a variety of microorganisms including periodontal pathogens. Chitosan also has antibacterial properties and can form a protective film on the surface of the oral mucosa to prevent bacterial adhesion and invasion. Zinc acetate dihydrate can enhance the antibacterial ability. At the same time, zinc ions help repair the oral mucosa. These ingredients are combined to produce a synergistic antibacterial effect and can more effectively inhibit the growth and reproduction of periodontal pathogens. Artemisinin is a natural ingredient extracted from plants, chitosan is a natural polysaccharide, and modified porous corn starch is also a relatively safe material. They all have good biocompatibility in the oral environment, so that the prepared balance agent is less irritating to the oral tissue during use and is not easy to cause adverse reactions such as allergies. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 The SEM images of corn starch and modified porous corn starch provided by the present invention; Figure 2 This is a comparative diagram of the release of the drug provided by the present invention in simulated saliva. 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 Ⅰ. Pour 0.1 g of zinc acetate dihydrate into 5 mL of deionized water and stir until completely dissolved to prepare solution A; Ⅱ. Pour 0.2 g of artemisinin into 3.5 mL of glacial acetic acid solution and stir until completely dissolved to prepare solution B; III. Pour 0.1 g of chitosan into 40 mL of deionized water, stir at room temperature for 5 min, then add 2.4 g of L-ascorbic acid, continue stirring for 30 min, and heat in a 70°C water bath for 10 min to obtain solution C; IV. Add solution A and solution B to solution C, continue heating in a water bath for 6 hours, add 6.5 g of modified porous corn starch, stir evenly, pour into a homogenizer and homogenize for 8 minutes to obtain a periodontal pathogen inhibition balancer; Wherein, the preparation method of modified porous corn starch is: (1) Pour 15 g corn starch into 80 mL of disodium hydrogen phosphate buffer solution with a pH value of 5-6, then place it in a 45°C water bath and heat it at a constant temperature for 20 min; add a composite enzyme consisting of α-amylase and saccharifying enzyme in a mass ratio of 1.2:0.7, place it in a 40°C constant temperature shaker, and react at a speed of 100 r / min for 8 h; after the reaction is completed, add 3.5 mL of NaOH solution with a mass fraction of 3-5%, filter it, wash the precipitate with distilled water, then place it in a 45°C drying oven to dry to constant weight, and finally grind it through a 0.15 mm mesh sieve to obtain porous corn starch; (2) The porous corn starch and distilled water in a mass ratio of 1:2.3 were stirred and mixed evenly to prepare a starch dispersion with a mass fraction of 30%; the mixture was placed in a constant temperature water bath and heated to 50°C, and stirred for 20 minutes, and then a NaOH solution with a mass fraction of 2-5% was added to adjust the pH value of the starch dispersion to 8.5; octenyl succinic anhydride with an addition amount of 3% of the mass of the porous starch and diluted 3 times with anhydrous ethanol was slowly added dropwise, and reacted for 3 hours in a water bath at 35°C. During the reaction, the pH value of the solution was controlled to be 8.2; after the reaction was completed, the mixture was filtered, and then the precipitate was washed to neutral with a 2-5% HCI solution, and then washed 3 times with anhydrous ethanol, and finally placed in a drying oven at 40°C for 18 hours, and then taken out, ground and sieved to obtain modified porous corn starch.

[0021] Example 2 Ⅰ. Pour 0.13 g of zinc acetate dihydrate into 5 mL of deionized water and stir until completely dissolved to prepare solution A; Ⅱ. Pour 0.22 g of artemisinin into 3.5 mL of glacial acetic acid solution and stir until completely dissolved to prepare solution B; III. Pour 0.14 g of chitosan into 40 mL of deionized water, stir at room temperature for 8 min, then add 2.8 g of L-ascorbic acid, continue stirring for 35 min, and heat in a 75°C water bath for 15 min to obtain solution C; IV. Add solution A and solution B to solution C, continue heating in a water bath for 7 hours, then add 8.5 g of modified porous corn starch, stir evenly, pour into a homogenizer and homogenize for 12 minutes to obtain a periodontal pathogen inhibitory balancer; Wherein, the preparation method of modified porous corn starch is: (1) Pour 18 g of corn starch into 90 mL of potassium dihydrogen phosphate buffer solution with a pH value of 5-6, then place it in a 50°C water bath and heat it at a constant temperature for 25 min; add a composite enzyme consisting of α-amylase and saccharifying enzyme in a mass ratio of 1.2:1, place it in a constant temperature shaker at 45°C, and react at a speed of 120 r / min for 9 h; after the reaction is completed, add 4 mL of NaOH solution with a mass fraction of 3-5%, filter it, wash the precipitate with distilled water, then place it in a drying oven at 48°C to dry it to constant weight, and finally grind it through a 0.20 mm mesh sieve to obtain porous corn starch; (2) The porous corn starch and distilled water in a mass ratio of 1:2.1 were stirred and mixed evenly to prepare a starch dispersion with a mass fraction of 32%; the mixture was placed in a constant temperature water bath and heated to 55°C, and stirred for 25 minutes, and then a NaOH solution with a mass fraction of 2-5% was added to adjust the pH value of the starch dispersion to 8.8; octenyl succinic anhydride with an addition amount of 4% of the mass of the porous starch and diluted 4 times with anhydrous ethanol was slowly added dropwise, and reacted in a water bath at 40°C for 3.5 hours. During the reaction, the pH value of the solution was controlled to be 8.4; after the reaction was completed, the mixture was filtered, and then the precipitate was washed to neutral with a 2-5% HCI solution, and then washed 4 times with anhydrous ethanol, and finally placed in a drying oven at 45°C for 21 hours, taken out, ground and sieved, and the modified porous corn starch was obtained.

[0022] Example 3 Ⅰ. Pour 0.16 g of zinc acetate dihydrate into 5 mL of deionized water and stir until completely dissolved to prepare solution A; Ⅱ. Pour 0.24 g of artemisinin into 3.5 mL of glacial acetic acid solution and stir until completely dissolved to prepare solution B; III. Pour 0.18 g of chitosan into 40 mL of deionized water, stir at room temperature for 10 min, then add 3.2 g of L-ascorbic acid, continue stirring for 40 min, and heat in a water bath at 80°C for 20 min to obtain solution C; IV. Add solution A and solution B to solution C, continue heating in a water bath for 8 hours, add 10 g of modified porous corn starch, stir evenly, pour into a homogenizer and homogenize for 15 minutes to obtain a periodontal pathogen inhibition balancer; Wherein, the preparation method of modified porous corn starch is: (1) Pour 20 g of corn starch into 100 mL of potassium dihydrogen phosphate buffer solution with a pH value of 5-6, then place it in a 55°C water bath and heat it at a constant temperature for 30 min; add a composite enzyme consisting of α-amylase and saccharifying enzyme in a mass ratio of 1.2:1.1, place it in a 50°C constant temperature shaker, and react at a speed of 140 r / min for 10 h; after the reaction is completed, add 5 mL of a Na0H solution with a mass fraction of 3-5%, filter it, wash the precipitate with distilled water, then place it in a 50°C drying oven to dry to constant weight, and finally grind it through a 0.25 mm mesh sieve to obtain porous corn starch; (2) The porous corn starch and distilled water in a mass ratio of 1:1.8 were stirred and mixed evenly to prepare a starch dispersion with a mass fraction of 35%; the mixture was placed in a constant temperature water bath and heated to 60°C, and stirred for 30 minutes, and then a NaOH solution with a mass fraction of 2-5% was added to adjust the pH value of the starch dispersion to 9.0; octenyl succinic anhydride with an addition amount of 5% of the mass of the porous starch and diluted 4 times with anhydrous ethanol was slowly added dropwise, and reacted in a water bath at 45°C for 4 hours. During the reaction, the pH value of the solution was controlled to be 8.6; after the reaction was completed, the mixture was filtered, and then the precipitate was washed to neutral with a 2-5% HCI solution, and then washed with anhydrous ethanol for 5 times, and finally placed in a drying oven at 50°C for 24 hours, taken out, ground and sieved, and the modified porous corn starch was obtained.

[0023] Comparative Example 1 Compared with Example 1, the difference is that the modified porous corn starch in Example 1 is replaced by corn starch; the rest remains unchanged.

[0024] Comparative Example 2 Compared with Example 1, the difference is that chitosan is not added; the rest remains unchanged.

[0025] Drug release in simulated saliva: Artificial saliva (SSF) was prepared with 0.4 g / L NaCl, 0.4 g / L KCl, 0.795 g / L CaCl2·2H20, 0.78 g / L NaH2PO4·2H20, 0.05 g / L Na2S·2H2O, 0.1 g urea and 10 U / mL a-amylase; 20 mg of the sample was weighed and placed in a dialysis bag (molecular cutoff value 3500 Da) containing 2 mL of artificial saliva, immersed in 80 mL SSF release medium, and incubated at 37 ° C in an oscillating shaker at 100 rpm for 48 h. During the incubation process, 1 mL of the release medium was taken out at regular intervals to measure the drug concentration, and an equal amount of fresh SSF was added to keep the volume of the release medium constant.

[0026] Antibacterial performance test: Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, and Tannerella forsythia were used as test bacteria to measure the antibacterial rate; the results are shown in Table 1.

[0027] like Figure 1 As shown, the surface of corn starch is smooth, while the surface of the modified porous corn starch is rough and has a large number of pore structures, which makes it easier for antibacterial ingredients to be adsorbed therein.

[0028] like Figure 2 As shown, the modified porous corn starch has significantly improved drug sustained-release performance compared to the untreated corn starch.

[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. A periodontal pathogenic bacteria inhibitory balance agent compounded with artemisinin components, characterized in that: The periodontal pathogenic bacteria inhibitor balancer mainly comprises modified porous corn starch, chitosan and artemisinin, and the preparation method thereof is as follows: Ⅰ. Pour zinc acetate dihydrate into deionized water and stir until completely dissolved to prepare solution A; Ⅱ. Pour artemisinin into glacial acetic acid solution and stir until completely dissolved to obtain solution B; III. Pour chitosan into deionized water, stir at room temperature, then add L-ascorbic acid, continue stirring, and heat in a water bath to obtain solution C; IV. Add solution A and solution B to solution C, continue heating in a water bath, add modified porous corn starch, stir evenly, and pour into a homogenizer for homogenization to obtain a periodontal pathogen inhibition balancer.

2. The periodontal pathogenic bacteria inhibitory balancing agent compounded with artemisinin components according to claim 1, characterized in that: The preparation method of the modified porous corn starch is: S1. Pour porous corn starch into distilled water, stir and mix evenly to prepare a starch dispersion with a mass fraction of 30%-35%; S2. Place the mixture in a constant temperature water bath and heat it to 50-60°C, stir for 20-30 minutes, then add 2-5% by mass NaOH solution to adjust the pH value of the starch dispersion to 8.5-9.0; S3, slowly dropwise add octenyl succinic anhydride diluted 3-4 times with anhydrous ethanol, and react in a water bath at 35-45°C for 3-4h; S4. After the reaction is completed, filter the mixture, wash the precipitate with 2-5% HCI solution until it is neutral, and then wash it with anhydrous ethanol for 3-5 times. Finally, dry it in a drying oven at 40-50°C for 18-24 hours, take it out, grind and sieve it to obtain modified porous corn starch.

3. The periodontal pathogenic bacteria inhibitory balancing agent compounded with artemisinin components according to claim 2, characterized in that: In the S1, the mass ratio of porous corn starch to distilled water is 1:(1.8-2.3).

4. The periodontal pathogenic bacteria inhibitory balancing agent compounded with artemisinin components according to claim 2, characterized in that: In S3, the added amount of octenyl succinic anhydride is 3%-5% of the mass of the porous starch, and during the reaction, the pH value of the solution is controlled to be 8.2-8.

6.

5. The periodontal pathogenic bacteria inhibitory balancing agent compounded with artemisinin components according to claim 2, characterized in that: The preparation method of the porous corn starch is: (1) Pour corn starch into the buffer solution and then place it in a water bath; (2) Add complex enzyme and place in a constant temperature shaker for reaction; (3) After the reaction is completed, add NaOH solution, filter and wash the precipitate with distilled water, then place it in a drying oven to dry to constant weight, and finally grind and sieve to obtain porous corn starch.

6. The periodontal pathogenic bacteria inhibitory balancing agent compounded with artemisinin components according to claim 5, characterized in that: In the above (1), the buffer solution is selected from any one of disodium hydrogen phosphate and potassium dihydrogen phosphate, and the pH value is 5-6.

7. The periodontal pathogenic bacteria inhibitory balancing agent compounded with artemisinin components according to claim 5, characterized in that: In the above (2), the complex enzyme is composed of α-amylase and saccharifying enzyme, and the mass ratio thereof is 1.2:(0.7-1.1); the water bath temperature is 45-55°C, and the constant temperature heating is performed for 20-30 minutes.

8. The artemisinin-containing periodontal pathogenic bacteria inhibitory balancing agent according to claim 1, characterized in that: In (2), the temperature of the constant temperature shaker is 40-50°C, and the reaction is carried out at a rotation speed of 100-140 r / min for 8-10 hours; in (3), the drying temperature is 45-50°C.

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