Application of cinnamon extract in preparation of porphyromonas gingivalis infection resisting medicine or oral cavity cleaning and nursing material

Abstract

The invention provides application of a cinnamon extract in preparation of a medicine for resisting porphyromonas gingivalis infection or an oral cavity cleaning and nursing material, and belongs to the technical field of traditional Chinese medicine extracts, and the use concentration of the cinnamon extract in the medicine is larger than or equal to 220 micrograms per milliliter; the action time is 10 to 40 minutes. The medicine is an external preparation, and the dosage forms of the medicine comprise gargle, gel and periodontal sustained-release agent. The oral cavity cleaning and nursing material comprises toothpaste, mouthwash or oral cavity spray. After the cinnamon extract VII stock solution acts on porphyromonas gingivalis with the concentration of 2 * 10 < 4 > cfu / mL for 30 minutes, the sterilization rate is greater than 99.9%; therefore, the cinnamon extract VII provided by the invention has a remarkable antibacterial effect on the porphyromonas gingivalis, can be applied to the preparation of a medicine for resisting the infection of the porphyromonas gingivalis or an oral cavity cleaning and nursing material, and has a good action effect.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine extracts, and in particular relates to application of a cinnamon extract in preparing a medicine for resisting Porphyromonas gingivalis infection or an oral cleaning and care product. Background Art

[0002] Porphyromonas gingivalis, a Gram-negative obligate anaerobe, is a key pathogenic microorganism in oral diseases such as chronic periodontitis and aggressive periodontitis. The virulence factors it secretes, such as gingipains and endotoxins, not only trigger periodontal inflammation and bone resorption but are also closely linked to the development and progression of systemic diseases such as cardiovascular disease, diabetes, and Alzheimer's disease through blood circulation or immune responses. Therefore, effectively inhibiting the growth and reproduction of P. gingivalis is of great significance for maintaining oral health and preventing systemic diseases.

[0003] Currently, clinical treatment for Porphyromonas gingivalis infection primarily relies on mechanical debridement and chemical antimicrobial drugs. Mechanical debridement, including supragingival scaling and subgingival curettage, is difficult to completely remove bacterial biofilms deep within the periodontal pockets. While chemical antimicrobial drugs such as metronidazole and amoxicillin can exert a certain antibacterial and bactericidal effect, long-term use can easily lead to bacterial resistance and may disrupt the balance of the oral microecology, causing dysbiosis. It can also cause adverse reactions such as gastrointestinal discomfort and allergic reactions. Furthermore, recent studies have found that Porphyromonas gingivalis in some patients has become resistant to traditional antibiotics, further increasing the difficulty of treatment.

[0004] Traditional Chinese medicine extracts, with their multi-component and multi-target properties, demonstrate unique advantages in the antibacterial field. Many traditional Chinese medicines contain active ingredients such as flavonoids, alkaloids, and terpenes, which can exert antibacterial effects through various mechanisms, including interfering with bacterial biofilm formation, inhibiting the expression of virulence factors, and regulating host immunity. Compared to chemical drugs, traditional Chinese medicine extracts have a lower risk of inducing resistance and exhibiting fewer adverse reactions. However, current research on traditional Chinese medicine extracts targeting Porphyromonas gingivalis is largely at the basic experimental stage, lacking a systematic and efficient extraction process and a clear mechanism of action, and lacking mature products with clinical application value. Summary of the Invention

[0005] In view of this, the object of the present invention is to provide a use of a cinnamon extract in the preparation of a drug for resisting Porphyromonas gingivalis infection; the specific cinnamon extract VII extracted and separated from cinnamon of the present invention has a significant effect of inhibiting and killing Porphyromonas gingivalis, and can be used to treat oral diseases caused by Porphyromonas gingivalis infection.

[0006] The present invention provides an application of cinnamon extract VII in preparing a medicine for resisting Porphyromonas gingivalis infection or an oral cleaning and care product, wherein the concentration of the cinnamon extract in the medicine is ≥220 μg / mL.

[0007] Preferably, the action time of the cinnamon extract VII is 10 to 40 minutes.

[0008] Preferably, the drug is an external preparation, and the dosage form of the drug includes a mouthwash, a gel, and a periodontal sustained-release agent.

[0009] Preferably, the oral cleaning and care products include toothpaste, mouthwash or oral spray.

[0010] Preferably, the cinnamon extract VII uses cinnamon as raw material, and is extracted by moving bed extraction to obtain a full-component extract; the obtained full-component extract is then separated by countercurrent chromatography, and the effluent at different time periods is collected and distilled to obtain a refined extract, and the refined extract that can kill Porphyromonas gingivalis is collected to obtain cinnamon extract VII.

[0011] Preferably, the cinnamon extract VII comprises coumarin, anethole, trans-cinnamaldehyde, 2'-methoxycinnamaldehyde and methyl cinnamate.

[0012] Preferably, the killing rate of the cinnamon extract VII on Porphyromonas gingivalis is ≥99.9%. Compared with the prior art, the present invention has the following beneficial effects: the cinnamon extract VII provided by the present invention has significant activity in killing Porphyromonas gingivalis; the present invention applies the cinnamon extract VII stock solution to 2×10 4 After 30 minutes of treatment with Porphyromonas gingivalis at a concentration of cfu / mL, the sterilization rate was greater than 99.9%, indicating that the cinnamon extract VII provided by the present invention has a significant antibacterial effect on Porphyromonas gingivalis, and can be used to prepare drugs or oral cleaning and care products for anti-Porphyromonas gingivalis infection, with good effects.

[0013] In addition, the cinnamon extract VII provided by the present invention has significant antibacterial effects on Escherichia coli, Staphylococcus aureus and Candida albicans. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 Total ion current diagram for component detection of cinnamon extract VIⅠ using LCMS: from top to bottom: positive mode, negative mode, and ultraviolet absorption diagram at a wavelength of 254nm. DETAILED DESCRIPTION

[0015] The present invention provides the use of cinnamon extract VII in the preparation of a medicament or oral hygiene product for treating Porphyromonas gingivalis infection. The cinnamon extract is used in the medicament at a concentration of ≥220 μg / mL, preferably 220 to 500 μg / mL. In the present invention, the duration of action of cinnamon extract VII is 10 to 40 minutes, preferably 20 to 35 minutes, and more preferably 30 minutes.

[0016] In the present invention, the drug is an external preparation, and the dosage form of the drug preferably includes a mouthwash, a gel, or a periodontal sustained-release agent. The present invention does not particularly limit the composition and preparation method of the gel or periodontal sustained-release agent, as long as the concentration and activity of cinnamon extract VII in the drug are ensured.

[0017] In the present invention, the oral hygiene product includes a toothpaste, mouthwash, or oral spray; the concentration of cinnamon extract VII in the oral hygiene product is preferably ≥ 220 μg / mL. The composition and preparation method of the toothpaste, mouthwash, or oral spray are not particularly limited in the present invention; conventional preparation methods in the art may be employed as long as the concentration and activity of cinnamon extract VII in the oral hygiene product are maintained.

[0018] In the present invention, the preparation method of cinnamon extract VII comprises the following steps:

[0019] 1) extracting cinnamon bark by a moving bed to obtain a full-component extract; 2) subjecting the full-component extract obtained in step 1) to countercurrent chromatography, and collecting effluents at different time periods; 3) distilling the collected effluents to obtain a refined extract, and collecting the refined extract having the effect of killing Porphyromonas gingivalis to obtain cinnamon bark extract VII.

[0020] In the present invention, the cinnamon bark is cinnamon twigs, cinnamon leaves, or cinnamon bark; cinnamon extract VII obtained from cinnamon twigs, cinnamon leaves, or cinnamon bark has similar properties; the cinnamon bark is preferably ground and then passed through a 20-40 mesh sieve. The structure of the moving bed and the moving bed extraction method are described in Patent 202411126616.3. The extraction solvent used in the present invention is an aqueous ethanol solution with a volume concentration of 50% to 60%, preferably 52% to 58%, and more preferably 55%. The extraction solvent is pumped at a rate of preferably 40 to 60 ml / min, more preferably 45 to 55 ml / min, and most preferably 50 ml / min. The mass-to-volume ratio of the cinnamon bark to the extraction solvent is preferably 1 g: (8-12) mL, preferably 1 g: (9-11) mL, and more preferably 1 g: 10 mL. After the moving bed extraction, the extract at the downstream terminal outlet is collected and concentrated to 1 / 3 of its original volume by reduced pressure to obtain a complete extract.

[0021] In the present invention, the obtained full-component extract is subjected to countercurrent chromatography, and the effluent is collected at different time periods. The apparatus used for countercurrent chromatography is described in Patent 202411086627.3. The countercurrent chromatography described in the present invention uses an ethanol-water solution with a volume concentration of 25% to 35% as the stationary phase, preferably 27% to 33%, and more preferably 30%; and n-hexane as the mobile phase. Specifically, the ethanol-water solution is mixed with the full-component extract to obtain a sample solution, ensuring that the volume concentration of ethanol in the sample solution is 25% to 35% (i.e., the sample is loaded with the stationary phase). After loading, the mobile phase is pumped in for countercurrent chromatography. In the present invention, the pumping rate of the mobile phase is preferably 5 to 10 ml / min, more preferably 7 to 9 ml / min, and most preferably 8 ml / min. During the countercurrent chromatography process, the sample components rapidly partition between the two phases. Due to differences in the partition coefficients, the different components flow out of the rotary joint at the outlet of the device along with the mobile phase. The effluent at different time periods is collected and labeled as component 1 to component 15.

[0022] In the present invention, the collected effluent is distilled to obtain a refined extract. The refined extract that has the ability to kill Porphyromonas gingivalis is collected to obtain Cinnamon Extract VII. In the present invention, the distillation temperature is 62-98°C, preferably 94-96°C, and more preferably 95°C, and the distillation time is 2-3 hours. In the present invention, the steam carries the components in the cinnamon and flows out after cooling through a condenser, obtaining refined extracts of various fractions. The refined extracts of the obtained fractions are tested for anti-Porphyromonas gingivalis activity, and the refined extract with anti-Porphyromonas gingivalis activity, i.e., the refined extract of Fraction 7, is collected to obtain Cinnamon Extract VII.

[0023] In the present invention, the cinnamon extract VII includes coumarin, anethole, trans-cinnamaldehyde, 2'-methoxycinnamaldehyde and methyl cinnamate, and also includes some unknown components.

[0024] The technical solutions provided by the present invention are described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0025] Example 1

[0026] Preparation and detection of active ingredients of cinnamon extract VIⅠ

[0027] In this embodiment, cassia bark from cinnamon bark is used as raw material, and the cassia bark is crushed and passed through a 20-40 mesh sieve.

[0028] 1) Moving bed extraction

[0029] The device used is described in patent 202411126616.3;

[0030] System construction: The fluid flow system is configured into 5 areas, numbered sequentially from area (5) to area (1), with area (5) as the upstream starting point and area (1) as the downstream end point. Each area (i) is provided with a liquid inlet hole ZI (i) and an outlet ZO (i), which are connected by pipes to form a liquid series passage. The system is configured with 5 tanks, 3 of which are used in this embodiment. An extraction tank with a loading and unloading port and a liquid inlet and outlet is placed in each area. The cinnamon raw material crushed to 20-40 mesh is loaded into 3 extraction tanks respectively, with each extraction tank containing 2 kg of cinnamon raw material. The loading and unloading port is closed, and the extraction tank is connected to the liquid inlet and outlet of the corresponding area.

[0031] Gradient extraction: ethanol with a volume concentration of 50% was selected as the extraction solvent. The extraction solvent was pumped into the upstream starting point liquid inlet ZI(f) at a flow rate of 50 ml / min, so that it flowed through the cinnamon bark raw material in the extraction tanks in areas (5), (4), and (3) in sequence. After 20 L of the predetermined volume of extraction solvent was pumped in, the pump was stopped and the extract at the downstream terminal outlet ZO(3) was collected.

[0032] The residue in the extraction tank in area (5) is unloaded, new raw materials are loaded, and the tank is moved to area (1); the tank in the original area (4) is moved to area (5), and the tank in the original area (3) is moved to area (4), and an extraction tank filled with unextracted cinnamon raw materials is installed in the original area (3) and connected;

[0033] The extraction solvent is pumped in again from the upstream starting point liquid inlet ZI(f) at a flow rate of 50 ml / min, so that it flows through the cinnamon raw material in the extraction tanks in areas (5), (4), and (3) in sequence. After pumping in 20 L of the predetermined volume of extraction solvent, the pump is stopped and the extract at the downstream terminal outlet ZO(3) is collected.

[0034] Pre-extract liquid treatment: The extract collected from the downstream terminal outlet ZO (1) is concentrated to 1 / 3 of the original volume by vacuum concentration to obtain a full-component extract rich in non-volatile components such as polyphenols and polysaccharides, which is set aside.

[0035] 2) Countercurrent chromatography separation stage

[0036] The device used is described in patent 202411086627.3;

[0037] Device preparation: Build a countercurrent chromatography device, ensure that the rotating shaft (20) is firmly connected to the rotating frame (10), and install 142 tubular containers (30) in series on the rotating frame (10), and place a magnetic stirring element (33) in each container (30). Set the end face of the container (30) close to the rotating shaft (20) as the feed liquid inlet (31), and the end face away from the rotating shaft (20) as the feed liquid outlet (32), completing the series connection between the containers. Install a magnetic field forming element (40) on one side of the rotating frame (10) so that its arc around the rotating frame (10) does not exceed 180 degrees. 0 The magnetic field can be generated by a permanent magnet or an electromagnet, and the inlet and outlet rotary joints as well as auxiliary equipment such as pumps and liquid storage tanks are connected.

[0038] Stationary phase filling and sample loading: using a heavy phase solution (ethanol-water solution) as the stationary phase and an immiscible light phase solution (n-hexane) as the mobile phase, the ethanol-water solution is mixed with the concentrated full-component extract to obtain a sample solution, ensuring that the volume concentration of ethanol in the sample solution is 30-35%; starting the rotating drum, rotating it at a constant speed of 500 rpm, pumping 600 ml of sample solution into the rotary joint at the inlet end of the equipment until the liquid flows out from the rotary joint at the outlet end, ensuring that all containers (30) are filled with the sample solution.

[0039] Sample separation: The mobile phase was pumped into the rotary joint at the inlet of the device at a rate of 8 ml / min. Under the action of the magnetic stirring element (33), the sample components were rapidly distributed between the two phases of solution. Due to the difference in distribution coefficients, the different components flowed out of the rotary joint at the outlet of the device along with the mobile phase. The effluent was collected in sections every 50 minutes and labeled as component 1, component 2, component 3 to component 15. The components obtained by countercurrent chromatography were separated by using a rotary evaporator to recover the solvent to obtain 15 separated components.

[0040] 3) Distillation stage

[0041] Raw material preparation: Transfer the separated fractions 1 to 15 to a distillation kettle respectively. Add 50 times the volume of water according to the characteristics of the fractions and the target composition to fully immerse the cinnamon fraction.

[0042] Distillation: Heat the still to 95°C and perform steam distillation. Control the distillation temperature and time (95°C, 2-3 hours) to ensure full extraction of volatile components and avoid component decomposition and energy waste. The resulting steam, carrying components from the cinnamon bark, flows out through a condenser after cooling, yielding a refined extract of components 1-15.

[0043] The refined extracts of the above components 1 to 15 were tested for their anti-Porphyromonas gingivalis activity, and it was found that component 7 had a significant effect of killing Porphyromonas gingivalis. The refined extract of component 7 was named cinnamon extract VIⅠ.

[0044] Component Analysis of Cinnamon Extract VIⅠ by LCMS

[0045] The equipment and reagents used are shown in Tables 1 and 2.

[0046] Table 1 Equipment information

[0047]

[0048] Table 2 Reagent information

[0049]

[0050] Experimental process

[0051] Sample: Cinnamon Extract VIⅠ obtained from the above extraction, cinnamon bark 76793003, sample number WT202504014-01.

[0052] Sample preparation: Centrifuge cinnamon extract VIⅠ directly and take the supernatant as the test solution.

[0053] Instrument parameters and testing conditions

[0054] Chromatographic column: Waters ACQUITY UPLC HSS T3 2.1×100mm×1.8um (LC-226) Mobile phase: A-acetonitrile, B-0.1% formic acid solution

[0055] Table 3 Elution gradient

[0056] Time (min) 0 10 20 25 30 31 35 A％ 5 30 60 90 90 5 5

[0057] Flow rate: 0.3 mL / min Wavelength: 254 nm, full wavelength scan Column temperature: 35°C

[0058] Injection volume: 1 μL

[0059] Ion source: ESI nozzle voltage: 4kV / 3.5kV

[0060] Sheath gas temperature: 350℃ Sheath gas flow rate: 11L / min Drying gas flow rate: 8L / min

[0061] Drying gas temperature: 325℃

[0062] Scan mode: Full scan

[0063] Primary scanning range: m / z 50~1700

[0064] Cone voltage: 100V

[0065] Primary scanning range: m / z 50~1700

[0066] Collision energy gradient: 10, 20, 40 Ev

[0067] Test results such as Figure 1 shown.

[0068] Data Processing: The collected raw data were imported into Qualitative Analysis 10.0 software. Using its wizard settings and method templates, an unknown compound identification process was established, and peak extraction was performed on the raw data. Characteristic peaks in the samples were analyzed, and possible molecular formulas were fitted using the extracted molecular ion chromatographic peaks and isotopic peaks. Primary information and secondary fragments were matched against the PCDL secondary database and the cinnamon plant library, filtering out results with mass deviations greater than 7.5 ppm and matching scores less than 70. Simultaneously, secondary fragment results were matched using the online database SIRIUS version 5.8.5. Compounds with matching scores greater than 70 and FingerIDs less than 50 were selected to obtain the final test results, as shown in Table 4.

[0069] Table 4 LCMS analysis results of cinnamon extract VIⅠ

[0070]

[0071] 1: * indicates that the peak may contain a peak.

[0072] 2: The results shown in the notes for PCDL secondary library and SIRIUS online library are all secondary fragment matching results, and those shown for plant library matching are all primary plant component library matching;

[0073] 3: The retention times in the table are based on the 254 nm chromatogram in the positive mode. The retention times in the negative mode may deviate by 0.1-0.3 minutes due to the acquisition time interval, but this does not affect the judgment.

[0074] 4: “-” means there is no matching result, and the possible molecular weight of the peak is only speculated;

[0075] A total of 14 characteristic peaks were analyzed in the sample, and 3 peaks were almost not ionized and could not be identified;

[0076] Example 2

[0077] Determination of the anti-Porphyromonas gingivalis activity of the cinnamon extract VII prepared in Example 1

[0078] Test items: sterilization test;

[0079] Sample arrival date: April 27, 2025;

[0080] Testing completion date: May 19, 2025.

[0081] Test strain: Porphyromonas gingivalis ATCC 33277

[0082] Neutralizer: D / E neutralizing broth

[0083] method

[0084] 1. Inspection basis: WS / T650-2019 "Evaluation Methods for Antibacterial and Bacteriostatic Effects", sterilization rate standard requirement: >90%.

[0085] 2. Neutralizer identification test: The neutralizer used is D / E neutralization broth.

[0086] Action concentration: stock solution; Action time: 10min,

[0087] The experiment was repeated three times, and the test strain was Porphyromonas gingivalis ATCC33277.

[0088] result

[0089] 1. Neutralizer identification test

[0090] Group 1: Add 0.1 mL of bacterial suspension to 5.0 mL of neutralizer and mix for 10 min. Take 0.5 mL of the mixture and add 4.5 mL of neutralizer. Dilute 10-fold with neutralizer and take 1.0 mL for culture and counting.

[0091] Group 2: 0.1 mL of bacterial suspension was added to 5.0 mL of neutralization product, mixed for 10 min, 0.5 mL of the mixture was added to 4.5 mL of neutralization product, diluted 10-fold with the neutralization product, and 1.0 mL was aspirated for culture and counting.

[0092] Group 3: 0.1 mL of bacterial suspension was added to 5.0 mL of PBS and mixed for 10 min. 0.5 mL of the mixture was added to 4.5 mL of PBS, diluted 10-fold with PBS, and 1.0 mL was cultured and counted.

[0093] Group 4: 1.0 mL each of diluent and neutralizer was inoculated into the plate, and 1.0 mL was aspirated for culture and counting.

[0094] Table 5 Neutralizer identification test results

[0095]

[0096] Groups 1, 2, and 3 had similar amounts of test bacteria growing, with a bacterial count of 1×10 4 CFU / mL~9×10 4 The difference in colony count between groups was 3.2% (<15%). Group 4 showed no bacterial growth and the neutralizer was qualified.

[0097] 2. Killing effect on test bacteria

[0098] 100 μL of bacterial suspension was added to 5 mL of sample solution. After the specified time, 0.5 mL was added to 4.5 mL of the corresponding neutralizer. After 10 minutes of incubation, appropriate dilution was performed. 1.0 mL of the appropriate dilution was then sampled for viable bacterial culture counts. A control sample was also prepared. The results are detailed in Table 6.

[0099] Table 6 Sterilization results of samples on test bacteria

[0100]

[0101] IV. Conclusion

[0102] The results of three repeated tests showed that the sterilization rates of the sample solution against Porphyromonas gingivalis were 52%, 91% and >99.9% at action times of 10 min, 20 min and 30 min, respectively. The sample had an antibacterial effect against Porphyromonas gingivalis.

[0103] Example 3

[0104] Determination of the activity of cinnamon extract VII prepared in Example 1 against Escherichia coli, Staphylococcus aureus and Candida albicans

[0105] Escherichia coli antibacterial activity assay

[0106] 1. Sample name: Cinnamon Extract VII

[0107] 2. Test strain name, strain number, providing institution, and culture generation: Escherichia coli, 8099, Guangdong Huankai Microbiology Technology Co., Ltd., 7th generation;

[0108] 3. Culture medium and others: nutrient agar medium, phosphate buffer (0.03 mol / L pH 7.2), neutralizer (3.9% D / E neutralizing broth);

[0109] 4.Instrument name / model / number: Constant temperature incubator / DH6000BII / L-516, electronic balance / SQP / L-326, electron microscope / BM2100 / L-511, vertical steam pressure sterilizer / LDZX-60KSB / L-484, constant temperature water bath / DK-98-IIA / L-515, biological safety cabinet / BSC-1500HA2-X / L-499.

[0110] method

[0111] 1.Testing basis: Appendix C of GB15979-2002 Hygiene Standard for Disposable Sanitary Products;

[0112] 2. The test was repeated three times at a temperature of 20°C ± 1°C.

[0113] 3. Preparation of E. coli suspension: Dilute to the desired concentration with phosphate buffer (0.03 mol / L, pH 7.2);

[0114] 4. Escherichia coli sterilization test: Cinnamon extract VII solution was used to treat Escherichia coli in suspension.

[0115] result

[0116] The test was repeated three times at a temperature of 20°C ± 1°C. The cinnamon extract VII was used as a stock solution and the exposure time was 2 minutes. The average bactericidal rate against Escherichia coli was 93.06%.

[0117] Table 7 Results of Escherichia coli sterilization test

[0118]

[0119]

[0120] Note: No bacteria grew in the negative control group.

[0121] in conclusion

[0122] The test was repeated three times at a temperature of 20°C ± 1°C. Cinnamon Extract VII, used as a stock solution and exposed for 2 minutes, had an average bactericidal rate of 93.06% against E. coli, meeting the requirements of Appendix C of GB15979-2002, "Hygienic Standard for Disposable Sanitary Products," indicating that the product has antibacterial properties.

[0123] Staphylococcus aureus antibacterial activity assay

[0124] 1. Sample name: Cinnamon Extract VII

[0125] 2. Test strain name, strain number, providing unit and culture generation: Staphylococcus aureus, ATCC6538, Guangdong Huankai Microbiology Technology Co., Ltd., 7th generation;

[0126] 3. Culture medium and others: nutrient agar medium, phosphate buffer (0.03 mol / L pH 7.2), neutralizer (3.9% D / E neutralizing broth);

[0127] 4.Instrument name / model / number: Constant temperature incubator / DH6000BII / L-516, electronic balance / SQP / L-326, electron microscope / BM2100 / L-511, vertical steam pressure sterilizer / LDZX-60KSB / L-484, constant temperature water bath / DK-98-IIA / L-515, biological safety cabinet / BSC-1500IIA2-X / L-499.

[0128] method

[0129] 1.Testing basis: Appendix C of GB15979-2002 Hygiene Standard for Disposable Sanitary Products;

[0130] 2. The test was repeated three times at a temperature of 20°C ± 1°C.

[0131] 3. Preparation of Staphylococcus aureus suspension: Dilute to the desired concentration with phosphate buffer (0.03 mol / L, pH 7.2);

[0132] 4. Staphylococcus aureus sterilization test: Use cinnamon extract VII solution to act on Staphylococcus aureus in suspension.

[0133] result

[0134] The test was repeated three times at a temperature of 20°C ± 1°C and an exposure time of 2 minutes. The average bactericidal rate against Staphylococcus aureus was 92.87%.

[0135] Table 8: Results of the bactericidal test on Staphylococcus aureus

[0136]

[0137] Note: No bacteria grew in the negative control group.

[0138] in conclusion

[0139] The test was repeated three times at a temperature of 20°C ± 1°C. The original solution, with an action time of 2 minutes, had an average bactericidal rate of 92.87% against Staphylococcus aureus, meeting the requirements of Appendix C of GB15979-2002, "Hygienic Standard for Disposable Sanitary Products," indicating that the product has antibacterial properties.

[0140] Antibacterial activity assay against Candida albicans

[0141] 1. Sample name: Cinnamon Extract VII

[0142] 2. Test strain name, strain number, providing unit and culture generation: Candida albicans, ATCC10231, Guangdong Huankai Microbiology Technology Co., Ltd., 7th generation;

[0143] 3. Culture medium and others: Sabouraud agar, phosphate buffer (0.03 mol / L pH 7.2), neutralizer (3.9% D / E neutralizing broth);

[0144] 4.Instrument name / model / number: Constant temperature incubator / DH6000BII / L-516, electronic balance / SQP / L-326, electron microscope / BM2100 / L-511, vertical steam pressure sterilizer / LDZX-60KSB / L-484, constant temperature water bath / DK-98-IIA / L-515, biological safety cabinet / BSC-1500IIA2-X / L-499.

[0145] method

[0146] 1.Testing basis: Appendix C of GB15979-2002 Hygiene Standard for Disposable Sanitary Products;

[0147] 2. The test was repeated three times at a temperature of 20°C ± 1°C.

[0148] 3. Preparation of Candida albicans suspension: dilute to the desired concentration with phosphate buffer (0.03 mol / L, pH 7.2);

[0149] 4. Candida albicans bactericidal test: Cinnamon extract VII solution was used to act on Candida albicans in suspension.

[0150] result

[0151] The test was repeated three times at a temperature of 20°C ± 1°C. The average bactericidal rate of the original solution of cinnamon extract VII against Candida albicans was 92.40% after an action time of 2 minutes.

[0152] Table 9: Candida albicans sterilization test results

[0153]

[0154] Note: No bacteria grew in the negative control group.

[0155] in conclusion

[0156] The test was repeated three times at a temperature of 20°C ± 1°C. Cinnamon Extract VII, as a stock solution, had an average bactericidal rate of 92.40% against Candida albicans after a 2-minute exposure time, meeting the requirements of Appendix C of GB15979-2002, "Hygienic Standard for Disposable Sanitary Products," demonstrating the product's antibacterial properties.

[0157] Example 4

[0158] Acute oral toxicity test of cinnamon extract VII

[0159] According to the single maximum limit test in 2.3.1 of Technical Specifications for Disinfection (2002 edition) [20 animals (half male and half female) were gavaged with a dose of 5000 mg / kg body weight, and there was no death within 14 days, so it can be determined that the LD50 is greater than 5000 mg / kg body weight], the test results were evaluated and determined in accordance with 2.3.1.6 of 2.3.1 of Technical Specifications for Disinfection (2002 edition), and the results of the acute oral toxicity test of the test substance in mice were practically non-toxic.

[0160] As shown in the above examples, the cinnamon extract VII provided by the present invention has significant activity against Porphyromonas gingivalis (30 minutes, a sterilization rate greater than 99.9%). Cinnamon extract VII can be used to prepare drugs or oral hygiene products for treating P. gingivalis infection, with good efficacy. Furthermore, cinnamon extract VII has significant antibacterial effects against Escherichia coli, Staphylococcus aureus, and Candida albicans, and can also be used in the preparation of daily chemical products or cleaning and disinfection products.

[0161] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A use of cinnamon extract VII in the preparation of a drug or oral cleaning and care product for resisting Porphyromonas gingivalis infection, characterized in that: The cinnamon extract is used in the medicine at a concentration of ≥220 μg / mL.

2. The use according to claim 1, characterized in that The action time of the cinnamon extract VII is 10 to 40 minutes.

3. The use according to claim 1 or 2, characterized in that The medicine is an external preparation, and the dosage forms of the medicine include gargle, gel, and periodontal sustained-release agent.

4. The use according to claim 1 or 2, characterized in that The oral cleaning and care products include toothpaste, mouthwash or oral spray.

5. The use according to claim 1 or 2, characterized in that: The cinnamon extract VII uses cinnamon as a raw material, and is extracted by a moving bed to obtain a full-component extract; the obtained full-component extract is then separated by countercurrent chromatography, and the effluents at different time periods are collected and distilled to obtain a refined extract; the refined extract that can kill Porphyromonas gingivalis is collected to obtain the cinnamon extract VII.

6. The use according to claim 5, characterized in that The cinnamon extract VII includes coumarin, anethole, trans-cinnamaldehyde, 2'-methoxycinnamaldehyde and methyl cinnamate.

7. The use according to claim 6, characterized in that The killing rate of the cinnamon extract VII on Porphyromonas gingivalis is ≥99.9%.

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