Composite disinfection composition and application thereof in targeted disinfection of oral mucosa drug-resistant bacteria based on PDT technology

Abstract

The invention relates to the technical field of oral cavity disinfection, in particular to a compound disinfection composition and application thereof to targeted disinfection in oral mucosa drug-resistant bacteria based on a PDT technology, targeted disinfection on the drug-resistant bacteria on the oral mucosa is achieved through cooperation of the compound disinfection composition and the PDT photodynamic technology of specific-section light waves, and the targeted disinfection effect on the drug-resistant bacteria on the oral mucosa is achieved. And the physical and non-antibiotic antibacterial scheme does not increase the resistance of drug-resistant bacteria.

Description

Technical Field

[0001] The present application relates to the technical field of oral disinfection, and more specifically, to a composite disinfection composition and its application in targeted disinfection of drug-resistant bacteria in oral mucosa based on PDT technology. Background Art

[0002] At present, antibacterial liquids and antibacterial creams with low-level quaternary ammonium salts, chlorhexidine, phenolic iodine and other ingredients as main ingredients can be found everywhere in the market.

[0003] In addition, although antibacterial products have been continuously updated in recent years, it cannot be ignored that the drug resistance of drug-resistant bacteria has increased exponentially, and some drug-resistant bacteria have reached the point where no drugs can be used.

[0004] According to the monitoring report: 40% of the 20 drug-resistant bacteria have a resistance rate of more than 3%, among which Acinetobacter baumannii has a resistance rate of more than 82% to 16 antimicrobial drugs. The resistance rate of drug-resistant bacteria such as Pneumococcus increased from 11% in 2012 to more than 24% in 2022, an increase of 120%;

[0005] In summary, it is urgent to provide a method for disinfecting drug-resistant bacteria. Based on the limitations of existing methods for disinfecting drug-resistant bacteria, the present application provides a composite disinfection composition and its application in targeted disinfection of drug-resistant bacteria in oral mucosa based on PDT technology, which can reduce the resistance of drug-resistant bacteria to antibacterial drugs and disinfectants through physical, non-antibiotic antibacterial schemes. Summary of the invention

[0006] In order to achieve the elimination of drug-resistant bacteria, the present application specifically provides a composite disinfection composition and its application in targeted disinfection of drug-resistant bacteria in oral mucosa based on PDT technology. Targeted disinfection of pathogenic bacteria on the oral mucosa can be achieved by adopting components such as photosensitizers in combination with PDT photodynamic technology of a specific light wave segment. Physical, non-antibiotic antibacterial solutions will not increase the resistance of drug-resistant bacteria.

[0007] In the first aspect, the present application provides a composite disinfection composition, which is as follows:

[0008] The composite disinfection composition comprises the following components in parts by weight:

[0009] 5-15 parts of plant photosensitizer; 5-10 parts of disinfectant; 3-5 parts of surfactant;

[0010] The plant photosensitizer is prepared by photophobic anaerobic fermentation of loquat leaves, liquorice leaves and Centella asiatica.

[0011] By adopting the above technical scheme, the photosensitizer prepared with natural plants as raw materials, after being compounded with disinfectants and surfactants, can adhere to dental plaque or biofilm targets, and when irradiated with light, produce singlet oxygen and active free radicals, destroying the structure of microbial biomacromolecules such as phospholipids, nucleic acids and proteins or affecting their functions, thereby achieving targeted disinfection.

[0012] Preferably, the plant photosensitizer is prepared by the following preparation steps, and the specific process is as follows:

[0013] S1. Firstly, fresh loquat leaves, licorice leaves and Centella asiatica are sheared, and then the sheared products are immersed and roughly extracted with ethanol aqueous solution to obtain a preformed product, and ultrasonic conditions are applied during the crude extraction process;

[0014] S2, adding cellulase and pectinase to the preformed product after ultrasonic treatment in S1, and then performing anaerobic fermentation in the dark for 3-6 hours, and concentrating the obtained mixed fermentation liquid to a paste by reduced pressure to obtain the plant photosensitizer.

[0015] By adopting the above technical solution and the light-protected mixed anaerobic fermentation method, chlorophyll is more likely to lose the magnesium in the porphyrin ring under acidic conditions to become pheophytin a, and further degrade to form pheophytin a-hexyl ether derivative with a strong absorption peak, which is more conducive to disinfection and sterilization.

[0016] Preferably, the weight ratio of loquat leaves, licorice leaves and Centella asiatica is 1:(0.3-0.5):(0.1-0.3).

[0017] Preferably, the ultrasonic conditions are: power 300-500W, frequency 10-20KHz, intermittent treatment 3-5min, and intermittent time 10s.

[0018] Preferably, the conditions for the anaerobic fermentation are: fermentation temperature of 48-52° C., pH value of 6.0-6.5, wherein the additive for adjusting the pH is copper chloride.

[0019] By adopting the above technical scheme, the temperature, time and specific pH regulator of the above-mentioned light-proof anaerobic fermentation will control the fermentation process to produce a small amount of chlorophyllin, which is not easy to affect the effective amount of chlorophyll. At the same time, part of it will also form a complex with copper ions, which is beneficial to inhibit the self-oxidation of chlorophyll during subsequent use. Under light conditions, it can also synergize with chlorophyll for sterilization.

[0020] Preferably, the surfactant is selected from amphoteric surfactants of benzalkonium bromide cationic type and / or amino acid type.

[0021] Preferably, the invention further comprises 10-30 parts of a thickener, wherein the thickener is nano-sized titanium dioxide.

[0022] Preferably, the mixture further comprises 10-30 parts of a solvent, wherein the solvent is water and / or ethanol.

[0023] Preferably, the composite disinfection composition is composed of the following components in parts by weight:

[0024] 5-15 parts of plant photosensitizer, 5-10 parts of disinfectant, 3-5 parts of surfactant, 0-30 parts of thickener, and 0-30 parts of solvent.

[0025] By adopting the above technical solution, targeted disinfection is achieved through the combined use of PDT + natural photosensitizer + bactericide, which provides a new path for the treatment of oral drug-resistant bacteria and effectively extends the application of PDT technology in the oral cavity within the industry. In addition, since the photosensitizer raw material used is plant, it also has the advantages of low toxicity, small side effects, high targeting, and easy absorption by human tissues.

[0026] In the second aspect, the present application provides an application of a composite disinfection composition for targeted disinfection of resistant bacteria in oral mucosa based on PDT technology, and the application products include capsules, tablets, solid beverages, suppositories, ointments, and gels.

[0027] Preferably, the application method of the composite disinfection composition is as follows: first gargle with a disinfection product containing any of the composite disinfection compositions described above, and then irradiate the oral area with a composite light source, so as to carry out targeted disinfection of drug-resistant bacteria, and the treatment time is relatively short, which can effectively reduce the operation time.

[0028] By adopting the above technical scheme, the above composite disinfection composition can be used for targeted disinfection of drug-resistant bacteria in oral mucosa based on PDT technology, and can also be prepared into capsules, tablets, solid beverages, suppositories, ointments, and gel products by compounding other components or changing the preparation method;

[0029] In addition, the above-mentioned specific photosensitizers can also be irradiated with composite light sources, which effectively overcomes the cognitive barrier of the traditional concept that only UVC253.7 can sterilize and disinfect. The use of composite light sources can effectively improve the combined effect and better kill bacteria and their survival barriers, achieving targeted destruction of various drug-resistant bacteria, and coordinating with different photosensitizers and bactericides to achieve targeted disinfection and maximize the disinfection effect.

[0030] In summary, this application has the following beneficial effects:

[0031] 1. The composite disinfection composition in the present application uses natural plants as the source of photosensitizers, and is compounded with disinfectants and surfactants. After adhering to dental plaque or biofilm targets, it can produce singlet oxygen and active free radicals when irradiated with light, and react with microbial phospholipids, nucleic acids, proteins and other biomacromolecules to damage the microbial structure or affect its function, ultimately achieving the purpose of disinfection;

[0032] 2. The dark-protected mixed anaerobic fermentation method in the present application makes it easy for chlorophyll to lose the magnesium in the porphyrin ring under acidic conditions to become pheophytin a, and further degrade to form pheophytin a-hexyl ether derivative, which has a strong absorption peak and a very high singlet oxygen yield;

[0033] In addition, thanks to the temperature, time and specific pH regulator of the light-proof anaerobic fermentation, only a small amount of chlorophyllin will be produced during the fermentation process, and some of it will form a complex with copper ions, which is beneficial to inhibit the self-oxidation of chlorophyll and can also cooperate with chlorophyll in sterilization under light conditions.

[0034] 3. The present application uses U light sources of different wavelengths to make a composite light source, which can effectively improve the combined effect and better kill bacteria and their survival barriers, achieve targeted destruction of various drug-resistant bacteria, and achieve targeted disinfection and maximize the disinfection effect by cooperating with different photosensitizers and bactericides. The processing time is relatively short, which can effectively reduce the patient's surgery and pain time;

[0035] 4. This application can use a composite light source and a customizable multi-band lamp bead integrated core sheet light emitting device. Its design provides a good example for expanding new thinking for PDT applications. Combined with a specific photosensitizer, it effectively overcomes the traditional concept that only UVC253.7 can sterilize and disinfect;

[0036] 5. This application effectively fills the gap in oral applications of PDT technology in the industry through the composite formula of PDT + natural photosensitizer + bactericide. Through targeted disinfection, it provides a new solution for the treatment of oral drug-resistant bacteria. The photosensitizer used has low toxicity, few side effects, high targeting, and is easily absorbed by human tissues, thereby extending the application field of PDT technology and having extremely high application value. DETAILED DESCRIPTION

[0037] The present application is further described in detail below with reference to the embodiments.

[0038] Preparation Example 1-5

[0039] A plant photosensitizer, the weight ratio of its raw material components is shown in the following table, and it is prepared by the following preparation steps, and its specific process is as follows:

[0040] S1. First, fresh loquat leaves, licorice leaves and Centella asiatica are sheared according to the corresponding weight ratio, and then the sheared materials are soaked and roughly extracted with 70% ethanol aqueous solution at a solid-liquid ratio of 1:1 to obtain a preform;

[0041] Ultrasonic conditions were applied during the immersion and extraction process, and the ultrasonic conditions were controlled as follows: power 300 W, frequency 10 KHz, intermittent treatment 3 min, and intermittent time 10 s;

[0042] S2, adding cellulase and pectinase to the preformed product after ultrasonic treatment in S1, and then performing anaerobic fermentation in the dark for 3 hours, and concentrating the obtained mixed fermentation liquid to a paste under reduced pressure to obtain a plant photosensitizer;

[0043] The enzymatic activity of cellulase is 100,000 u / ml, and the added amount is 5‰ of the total weight of the preform;

[0044] The enzyme activity of pectinase is 50,000 u / ml, and the amount added is 3‰ of the total weight of the preform;

[0045] The conditions for anaerobic fermentation were: fermentation temperature 50°C, pH 6.2;

[0046] The additive for adjusting pH is copper chloride.

[0047] Table: Weight ratio of raw material components of plant photosensitizer in Preparation Examples 1-5

[0048]

[0049] Preparation Example 6-7

[0050] A plant photosensitizer, which is different from Preparation Example 1 in that the process conditions of its ultrasonic treatment are different, as shown in the following table:

[0051] Table: Ultrasonic conditions in Preparation Examples 6-7

[0052]

[0053] Preparation Examples 8-10

[0054] A plant photosensitizer, which is different from Preparation Example 1 in that the process conditions of its anaerobic fermentation are different, as shown in the following table:

[0055] Table: Anaerobic fermentation conditions in preparation examples 8-10

[0056]

[0057]

[0058] Performance testing

[0059] The composite disinfection compositions prepared in each embodiment and comparative example were respectively selected for bactericidal performance test, the application amount of the composite disinfection composition was 2.0 g, and the specific test steps and conditions were as follows:

[0060] Test conditions:

[0061] 1) Lighting condition I: Light dose 3.0 J / cm 2 , wavelength 640nm, irradiation time 3min;

[0062] Light condition II: or the total light dose is controlled at 3.0J / cm 2 , irradiated simultaneously by a composite light source with wavelengths of 640nm, 650nm, and 660nm for 3 minutes.

[0063] 2) Test strains: The ATCC series of standard strains provided by the International Pasteur as stipulated by WHO are treated with quaternary ammonium salt fungicides and are resistant to the multi-culture strain RE;

[0064] The initial bacterial strains are specifically selected from Escherichia coli (ATCC8099), Staphylococcus aureus (ATCC6538), and Candida albicans (ATCC90028), and the suffix RE of the complex strain is used to distinguish it.

[0065] 3) Test object: A simulated oral cavity cast with alginate water-powder impression material in accordance with the YY 91027-1999 standard, loaded with the above-mentioned strains to be tested, with the initial concentration of each strain being 1*10 12 CFU / mg, then test according to the above conditions, count the number of live bacteria after irradiation, and calculate the corresponding sterilization rate %.

[0066] Example

[0067] Examples 1-5

[0068] A composite disinfectant composition, the raw material components and dosages of which are shown in the following table (g), is prepared by dispersing and mixing at normal temperature and pressure:

[0069] The plant photosensitizer is prepared according to Preparation Example 1, the disinfectant is hydrogen peroxide, the surfactant is sodium cocoyl aminopropionate, the thickener is nano-scale titanium dioxide, and the solvent is 50% ethanol aqueous solution.

[0070] Table: Components and weights of the composite disinfection compositions in Examples 1-5 (g)

[0071]

[0072] Comparative Examples 1-3

[0073] A composite disinfection composition, which differs from Example 1 in that the raw material of the plant photosensitizer is replaced by an equal amount of a single plant component, as shown in the following table:

[0074] Table: Usage of plant photosensitizer raw materials in comparative examples 1-5

[0075]

[0076] Comparative Example 4

[0077] A composite disinfection composition, which is different from Example 1 in that the raw materials of the plant photosensitizer are the same, and only the preparation method is different, as follows:

[0078] S1. First, fresh loquat leaves, licorice leaves and Centella asiatica are sheared according to the corresponding weight ratio, and then the sheared materials are soaked and roughly extracted with 70% ethanol aqueous solution at a solid-liquid ratio of 1:1 to obtain a preform;

[0079] Ultrasonic conditions were applied during the immersion and extraction process, and the ultrasonic conditions were controlled as follows: power 300 W, frequency 10 KHz, intermittent treatment 3 min, and intermittent time 10 s;

[0080] S2, the preformed product after ultrasonic treatment in S1 is concentrated under reduced pressure to a paste, thereby obtaining a plant photosensitizer.

[0081] The composite disinfectant compositions in the above Examples 1-5 and Comparative Examples 1-4 were selected and their anti-inflammatory properties against different bacterial species were tested according to the above test steps and standards. The average values ​​of the test results were recorded in the following table.

[0082] Table: Performance test of Examples 1-5 and Comparative Examples 1-4

[0083]

[0084] As can be seen from the above table, the composite disinfection compositions prepared in Examples 1-5 all have excellent anti-inflammatory effects, and the sterilization rates of light I are all ≥95.0%, and the sterilization rates of light II are all ≥99.0%, which are improved to varying degrees compared with Comparative Examples 1-4. It can be seen that the plant photosensitizer prepared by special raw materials and processes in this application has a significant effect in terms of antibacterial performance;

[0085] Photosensitizers, disinfectants and surfactants, after adhering to dental plaque or biofilm targets, can generate singlet oxygen and active free radicals when exposed to light, which react with microbial biomacromolecules such as phospholipids, nucleic acids and proteins, damaging the microbial structure or affecting its function, and then achieving sterilization;

[0086] In addition, combined with the corresponding test data of Comparative Examples 1-4, it can be concluded that only the plant photosensitizer obtained by the combination of loquat leaves, licorice leaves and Centella asiatica and anaerobic fermentation has the corresponding effect. Further combined with the data, the principle may be as follows:

[0087] 1) The loquat leaves, licorice leaves and Centella asiatica components used not only have a large amount of chlorophyll components, but also have a certain compatibility effect, so they can effectively play a synergistic bactericidal role;

[0088] The specific theoretical basis can be found in Example 1 and Comparative Examples 1-3, that is, the sterilization rate of the groups using each component alone is ≤ 75%, which can only be regarded as having a certain antibacterial effect;

[0089] In addition, the intermittent ultrasonic treatment and mixed fermentation during the preparation process effectively avoid the high temperature conditions caused by heating and high speed, thereby achieving the retention of effective substances;

[0090] 2) The dark-protected mixed anaerobic fermentation method in the present application makes it easy for chlorophyll to lose the magnesium in the porphyrin ring under acidic conditions to become pheophytin a, and further degrade to form pheophytin a-hexyl ether derivative, which has a strong absorption peak and a very high singlet oxygen yield;

[0091] The theoretical basis can be found in Example 1 and Comparative Example 4. Although the same components as those in Example 1 of the present application are used in Comparative Example 4, since it is only conventional immersion extraction, ultrasonic treatment and reduced pressure concentration, only chlorophyll is played a role, and the effect is poor due to the stability of chlorophyll.

[0092] 3) In addition, thanks to the temperature, time and specific pH regulator of the light-proof anaerobic fermentation, only a small amount of chlorophyllin will be produced during the fermentation process, and some of it will form a complex with copper ions, which is beneficial to inhibit the self-oxidation of chlorophyll and can also synergize with chlorophyll to sterilize under light conditions.

[0093] It can also be seen from Examples 1-5 that the composite disinfection composition is preferably composed of the following components in parts by weight: 5-25 parts of plant photosensitizer, 5-10 parts of disinfectant, 3-5 parts of surfactant, 0-30 parts of thickener, and 0-30 parts of solvent. The performance fluctuation is small within this range.

[0094] The dosage of the thickener and the solvent can be adjusted and replaced according to the conventional method in the art according to the actual adhesion capacity requirements, but the overall adhesion capacity should not be too weak. Since the test conditions in this application are simulated oral cavity, the shedding phenomenon of the composite disinfection composition caused by gravity will also have a slight effect on the final performance, and when the thickener is nano-scale titanium dioxide, it is beneficial to further improve the performance.

[0095] Embodiment 6-14

[0096] A composite disinfection composition, which is different from Example 1 in that the plant photosensitizers used are used differently, and the specific corresponding relationships are shown in the following table.

[0097] Table: Comparison table of plant photosensitizer usage in Examples 6-14

[0098] Group Plant photosensitizer Example 6 Prepared from Preparation Example 2 Example 7 Prepared from Preparation Example 3 Example 8 Prepared by Preparation Example 4 Example 9 Prepared from Preparation Example 5 Example 10 Prepared from Preparation Example 6 Embodiment 11 Prepared from Preparation Example 7 Example 12 Prepared from Preparation Example 8 Example 13 Prepared from Preparation Example 9 Embodiment 14 Prepared from Preparation Example 10

[0099] The composite disinfectant compositions in Examples 6-14 were sampled and their anti-inflammatory properties against different bacterial species were tested according to the above test steps and standards. The average values ​​of the test results were recorded in the following table.

[0100] Table: Performance Tests of Examples 6-14

[0101]

[0102] It can be seen from the above table that the composite disinfection compositions prepared in Examples 6-14 all have excellent anti-inflammatory effects, and the sterilization rates of light I are all ≥90.0%, and the sterilization rates of light II are all ≥95.0%, which are improved to varying degrees compared with Example 1. It can be seen that the raw material ratio and process conditions of the present application will greatly affect the antibacterial effect of the plant photosensitizer, as follows;

[0103] It can be seen from Examples 1 and 6-9 that the weight ratio of loquat leaves, licorice leaves and Centella asiatica is preferably 1:(0.3-0.5):(0.1-0.3). Beyond this range, the performance decreases slightly. Preparation Example 3, i.e., Example 7, is the best example, and the compounding of the three is the most reasonable.

[0104] It can be seen from Examples 1, 10-11 that the preferred process conditions for ultrasonic treatment are as follows:

[0105] The power is 300-500W, the frequency is 10-20KHz, the intermittent treatment is 3-5min, and the intermittent time is 10s. Under the same treatment time, the intermittent treatment method is more conducive to the stability and acquisition of chlorophyll, and reduces the high temperature effect caused by continuous stirring. Refer to Example 10 and Example 11.

[0106] It can be seen from Examples 1, 12-14 that the preferred process conditions for anaerobic fermentation are as follows:

[0107] The fermentation temperature is 48-52°C and the pH value is 6.0-6.5. The additive for adjusting the pH is copper chloride. Since chlorophyll can lose the magnesium in the porphyrin ring and further degrade to form a-hexyl ether derivative of pheophytin with a strong absorption peak and high singlet oxygen yield, magnesium chloride is not recommended.

[0108] In addition, thanks to the temperature, time, pH and copper chloride selection of the light-proof anaerobic fermentation, only a small amount of chlorophyllin will be produced during the fermentation process, and some of it will form a complex with copper ions, which is beneficial to inhibit the self-oxidation of chlorophyll. Under light conditions, it can also synergize with chlorophyll for sterilization.

[0109] This specific embodiment is merely an explanation of the present application and is not a limitation of the present application. After reading this specification, those skilled in the art may make modifications to the present embodiment without any creative contribution as needed, but such modifications are protected by the patent law as long as they are within the scope of the claims of the present application.

Claims

1. A composite disinfectant composition, characterized in that: The composition comprises the following components in parts by weight: 5-15 parts of plant photosensitizer; 5-10 parts of disinfectant; 3-5 parts of surfactant; The plant photosensitizer is prepared by photophobic anaerobic fermentation of loquat leaves, liquorice leaves and Centella asiatica.

2. The composite disinfectant composition according to claim 1, characterized in that The plant photosensitizer is prepared by the following preparation steps, and the specific process is as follows: S1. Firstly, fresh loquat leaves, licorice leaves and Centella asiatica are sheared, and then the sheared products are immersed and roughly extracted with ethanol aqueous solution to obtain a preformed product, and ultrasonic conditions are applied during the crude extraction process; S2, adding cellulase and pectinase to the preformed product after ultrasonic treatment in S1, and then performing anaerobic fermentation in the dark for 3-6 hours, and concentrating the obtained mixed fermentation liquid to a paste by reduced pressure to obtain the plant photosensitizer.

3. The composite disinfectant composition according to claim 2, characterized in that The weight ratio of loquat leaves, liquorice leaves and Centella asiatica is 1:(0.3-0.5):(0.1-0.3).

4. The composite disinfectant composition according to claim 2, characterized in that The ultrasonic conditions are: power 300-500W, frequency 10-20KHz, intermittent treatment 3-5min, and intermittent time 10s.

5. The composite disinfectant composition according to claim 2, characterized in that: The conditions of the anaerobic fermentation are: fermentation temperature 48-52° C., pH value 6.0-6.5, wherein the additive for adjusting pH is cupric chloride.

6. The composite disinfectant composition according to claim 1, characterized in that The surfactant is selected from amphoteric surfactants of benzalkonium bromide cation type and / or amino acid type.

7. The composite disinfectant composition according to claim 1, characterized in that The invention also comprises 10-30 parts of a thickener, wherein the thickener is nano-sized titanium dioxide.

8. The composite disinfectant composition according to claim 1, characterized in that The invention also comprises 10-30 parts of a solvent, which is water and / or ethanol.

9. The composite disinfection composition according to any one of claims 1 to 8, characterized in that: The invention is composed of the following components in parts by weight: 5-15 parts of plant photosensitizer, 5-10 parts of disinfectant, 3-5 parts of surfactant, 0-30 parts of thickener and 0-30 parts of solvent.

10. Application of a composite disinfection composition for targeted disinfection of drug-resistant bacteria in oral mucosa based on PDT technology, characterized in that: The application method is as follows: The drug-resistant bacteria can be targetedly disinfected by first gargling the disinfection product containing the composite disinfection composition according to any one of claims 1 to 9, and then irradiating the oral area with a composite light source.

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