Antibacterial composition, antibacterial composition ointment as well as preparation method and application of antibacterial composition ointment

Abstract

The invention relates to the technical field of Chinese herbal medicines, and discloses an antibacterial composition, an antibacterial composition ointment as well as a preparation method and application thereof, and the antibacterial composition is prepared from the following components in parts by mass: 12 to 25 parts of fructus gardeniae extract and 6 to 12 parts of cortex cinnamomi extract. According to the antibacterial composition and the antibacterial composition ointment provided by the invention, the two traditional Chinese medicine extracts of the fructus gardeniae and the cortex cinnamomi are selected aiming at the main pathogenic bacteria of the skin tinea of cats and dogs, and the two traditional Chinese medicine extracts play a synergistic antibacterial effect, so that the effect of solving the skin tinea of cats and dogs is improved. The product is high in safety and has a good application prospect.

Description

Technical Field

[0001] The present invention relates to the technical field of Chinese herbal medicines, and in particular to an antibacterial composition, an antibacterial composition ointment, and a preparation method and application thereof. Background Art

[0002] Dermatophytosis is a common fungal skin disease of fur animals. Dermatophytes invade the skin, nails, hair and other parts of animals, causing symptoms such as hair loss, skin granulomas, and blisters. It has the characteristics of high incidence, strong infectivity, and easy recurrence. Common pets such as cats and dogs are susceptible to dermatophytosis. According to relevant statistics, the incidence rate of cats and dogs is about 6%. In addition, the disease is a zoonosis. People can also be infected through contact with sick animals, which may lead to public health problems. The pathogens that cause dermatophytosis in cats and dogs are mainly superficial fungi that parasitize on the superficial keratin tissue of the skin, such as Microsporum canis, Microsporum gypseum, and Trichophyton mentagrophytes. Among them, Microsporum canis is the main pathogen, accounting for 70% and 98% of dermatophyte infections in dogs and cats, respectively. At present, the products used in the market for the treatment of dermatophyte infections in dogs and cats are mainly chemical synthetic antifungal drugs, including polyenes, azoles, echinocandins, and pyrimidines, such as clotrimazole, terbinafine, and pitofen. Although these chemical drugs have good efficacy, they have large side effects, are prone to drug resistance, and have a high recurrence rate. Chinese herbal medicines have the characteristics of small side effects and resistance to drug resistance. In recent years, the development of Chinese herbal medicine products for the treatment of cat and dog dermatophytosis has received more and more attention. Summary of the invention

[0003] Purpose of the invention: In view of the problems existing in the prior art, the present invention provides an antibacterial composition, an antibacterial composition ointment, and a preparation method and application thereof. The present invention provides an efficient and safe solution for treating cat and dog dermatophyte Microsporum canis, a common pathogen that induces dermatophyte tinea in cats and dogs.

[0004] Technical solution: In the first aspect, the present invention provides an antibacterial composition, which is composed of 12-25 parts of gardenia extract and 6-12 parts of cinnamon extract in parts by mass.

[0005] Furthermore, the gardenia extract is specifically obtained by crushing the gardenia, adding an appropriate volume of ethanol with a concentration of 75% (v / v) or more, ultrasonically extracting, filtering, and concentrating under reduced pressure.

[0006] Furthermore, the cinnamon extract is obtained by crushing cinnamon, adding an appropriate volume of ethanol with a concentration of 75% (v / v) or more, ultrasonically extracting, filtering, and concentrating under reduced pressure.

[0007] Second aspect, the present invention provides an application of the antibacterial composition as described in any one of the above in the treatment of dermatophytosis in cats and dogs, wherein the pathogenic bacterium of the dermatophytosis in cats and dogs is Microsporum canis.

[0008] Third aspect, the present invention provides an antibacterial composition ointment, and the raw materials of the antibacterial composition ointment include the antibacterial composition as described in any one of the above and conventional excipients.

[0009] Fourth aspect, the present invention provides a preparation method of the antibacterial composition ointment as described in any one of the above. Take 12 - 25 parts of gardenia extract and 6 - 12 parts of cinnamon extract, add conventional excipients, and use the conventional preparation method of ointment to make an ointment.

[0010] Fifth aspect, the present invention provides an application of the antibacterial composition ointment as described in any one of the above in the treatment of dermatophytosis in cats and dogs, wherein the pathogenic bacterium of the dermatophytosis in cats and dogs is Microsporum canis.

[0011] Beneficial effects: Compared with the prior art, the specific beneficial effects of the present invention are as follows: The antibacterial composition and the antibacterial composition ointment provided by the present invention target the main pathogenic bacterium Microsporum canis of dermatophytosis in cats and dogs, and select two traditional Chinese medicine extracts, gardenia and cinnamon. The two play a synergistic antibacterial effect, improving the effect of solving dermatophytosis in cats and dogs. The product has high safety and good application prospects.

[0012] The present invention compares the in vitro antibacterial effects of the two extracts and their combination on Microsporum canis. The results show that both gardenia extract and cinnamon extract have a certain inhibitory effect on Microsporum canis. After the combination of the two, a synergistic antibacterial effect is shown. Specific embodiments

[0013] The present invention will be introduced in detail below in combination with the embodiments.

[0014] In the present invention, the used gardenia extract is obtained by crushing gardenia, adding 20 times the volume of 75% (v / v) ethanol, ultrasonic extracting at 30 °C for 1 h, filtering, concentrating the extract under reduced pressure, and freeze - drying.

[0015] The used cinnamon extract is obtained by crushing cinnamon, adding 20 times the volume of 75% (v / v) ethanol, ultrasonic extracting at 30 °C for 1 h, filtering, concentrating the extract under reduced pressure, and freeze - drying.

[0016] Embodiment 1: This embodiment provides an antibacterial composition, including: 12 parts of gardenia extract and 6 parts of cinnamon extract by mass. The above antibacterial composition is prepared into an ointment by using a conventional method and additives.

[0017] A preparation method of an ointment is provided herein. It should be noted that this method is only for reference and not all the methods covered by the present invention: The matrix prescription by mass fraction is: glycerol monostearate 5 parts, white petrolatum 20 parts, liquid paraffin 10 parts, glycerol 5 parts, ethylparaben 0.1 part. Heat the above glycerol monostearate, white petrolatum and liquid paraffin to 75 °C until melted, stir evenly to obtain the oil phase, and keep it warm for standby. Heat glycerol, ethylparaben and an appropriate amount of distilled water to 75 °C until dissolved, stir evenly to obtain the water phase, and keep it warm for standby. Slowly add the water phase to the oil phase under stirring, continue stirring until emulsification is complete to form a uniform cream matrix. Add the above antibacterial composition to the cream matrix, stir evenly, and homogenize at 8000 rpm for 5 minutes using a homogenizer to obtain the antibacterial composition ointment.

[0018] Embodiment 2: This embodiment provides an antibacterial composition, including by mass: 20 parts of gardenia extract and 8 parts of cinnamon extract.

[0019] Prepare the above antibacterial composition into an ointment by conventional methods and additives.

[0020] Embodiment 3: This embodiment provides an antibacterial composition, including by mass: 25 parts of gardenia extract and 12 parts of cinnamon extract.

[0021] Prepare the above antibacterial composition into an ointment by conventional methods and additives.

[0022] Inhibitory zone test: An experiment for measuring the antibacterial ability of different extracts by the agar plate diffusion method. Microsporum canis was purchased from Shanghai Yiji Biotech Co., Ltd., and the culture medium was comprehensive potato dextrose agar (CPDA). Mix 0.2 mol / L Na2HPO4 solution and KH2PO4 solution evenly at a volume ratio of 12.3:87.7 as the solvent to prepare the culture medium solution. Using DMSO as the solvent, prepare 100 mg / mL gardenia extract test solution and 100 mg / mL cinnamon extract test solution respectively. Cool the autoclaved culture medium to about 50 °C, accurately pipette 200 μL of Microsporum canis suspension (10 7(CFU / mL) was added to 20 mL of the above medium, mixed well and poured into a petri dish to make a bacteria-containing plate. Filter paper with a thickness of 0.15 cm was punched into circular pieces with a diameter of 6 mm, sterilized, and immersed in the above-mentioned Gardenia jasminoides Ellis extract and Cinnamomum cassia Presl extract test solutions for 2 hours respectively. After draining, it was pasted on the bacteria-containing plate with sterile forceps, and 4 medicated filter paper pieces were placed equidistantly in each petri dish and placed in an incubator. After culturing at 28 °C for 7 days, the diameter of the inhibition zone was measured. The determination was carried out in parallel 3 times, and the results are shown in Table 1. The judgment standard for the inhibition zone is: a diameter greater than 15 mm is the most sensitive type, 10 - 15 mm is the moderately sensitive type, 7 - 9 mm is the low-sensitive type, and a diameter less than 7 mm is the insensitive type. The test results show that the Gardenia jasminoides Ellis extract and Cinnamomum cassia Presl extract test solutions both have a certain inhibitory effect on Microsporum canis. On this basis, the next experiment of determining the minimum inhibitory concentration (MIC) by the liquid medium dilution method was carried out.

[0023]

[0024] Experiment on determining the minimum inhibitory concentration (MIC) by the liquid medium dilution method: Experiment on determining the minimum inhibitory concentration (MIC) of different extracts against Microsporum canis by the liquid medium dilution method. Using DMSO as the solvent, test solutions of Gardenia jasminoides Ellis extract and Cinnamomum cassia Presl extract with a concentration of 1000 mg / mL were prepared respectively. A row of test tubes was taken, and except for adding 9 mL of the medium solution to the first tube, 5 mL of the medium solution was added to each of the remaining tubes. 1 mL of the Gardenia jasminoides Ellis extract test solution in Example 1 was precisely pipetted into the first tube, ultrasonically mixed evenly to obtain a Gardenia jasminoides Ellis extract solution with a concentration of 100 mg / mL. 5 mL of the medicated medium solution was aspirated from the first tube to the second tube, ultrasonically mixed evenly and then 5 mL was aspirated to the third tube. After such continuous serial dilution, a series of Gardenia jasminoides Ellis extract solutions with concentrations of 3.12 - 100 mg / mL were obtained. According to the same method of operation, a series of Cinnamomum cassia Presl extract solutions with concentrations of 3.12 - 100 mg / mL were obtained respectively. After the two blank culture media and the above-mentioned test solutions were sterilized at high temperature, 20 μL of the Microsporum canis suspension in Example 1 was added respectively (no bacteria was added to the negative control solution), and it was placed in a shaking incubator for culturing at a rotation speed of 160 r / min. After culturing at 28 °C for 7 days, it was taken out and the growth of the strains in each tube was observed visually. The solution in the negative control tube was clear and transparent, indicating complete sterilization and no contamination during the culturing process. The solution in the positive control tube was turbid, indicating good growth of each strain. The lowest concentration of the test drug solution that was clear and transparent was taken as the minimum inhibitory concentration MIC of the extract against Microsporum canis. The test was carried out in parallel 3 times, and the results are shown in Table 2. On this basis, the next experiment of determining the antibacterial ability of different extract combinations by the liquid medium dilution method was carried out.

[0025]

[0026] Test for determining the antibacterial ability of different extract combinations by liquid medium dilution method: A two-factor and three-level experiment was conducted to investigate the inhibitory effect of different concentrations of extract combinations on Microsporum canis. Taking the MIC of each extract as the base value, 1 / 2, 1 / 4, and 1 / 8 of it were taken as the three levels. The experimental factor table is shown in Table 3, and the experimental table is shown in Table 4.

[0027]

[0028]

[0029] According to the designed extract concentrations of each sample in Table 4, using the test solution in the above liquid medium dilution experiment, 250 μL of Gardenia jasminoides Ellis extract solution and 120 μL of Cinnamomum cassia Presl extract solution were respectively pipetted into a 10 mL volumetric flask, added with culture medium solution to the scale, shaken evenly, and added to test tubes to obtain the test sample No. 1 in Table 4. Samples were taken and prepared in this way successively to obtain 9 sample solutions in Table 4. After two blank culture media and the above test solution were sterilized at high temperature, 20 μL of the Microsporum canis suspension prepared in the antibacterial circle test was added respectively (no bacteria were added to the positive control solution), and they were placed in a shaking incubator for cultivation at a rotation speed of 160 r / min. After culturing at 28 °C for 7 days, they were taken out and the growth of the strains in each tube was observed with the naked eye. The clear and transparent solution in the negative control tube indicated complete sterilization and no contamination during the cultivation process, and the turbid solution in the positive control tube indicated good growth of each strain. The clear sample solution was antibacterial positive "+", and the turbid sample solution was antibacterial negative "—". The test was determined in parallel three times, and the results are shown in Table 5.

[0030]

[0031] As can be seen from the results in Table 5, when the concentration of Gardenia jasminoides Ellis extract reaches 12 mg / mL and the concentration of Cinnamomum cassia Presl extract reaches 6 mg / mL, both show positive antibacterial results. It indicates that the critical concentrations of the combination of Gardenia jasminoides Ellis extract and Cinnamomum cassia Presl extract for antibacterial are 12 mg / mL and 6 mg / mL respectively. This result shows that the combination of Gardenia jasminoides Ellis extract and Cinnamomum cassia Presl extract has a significantly better inhibitory effect on Microsporum canis than either extract alone. At the same time, the minimum inhibitory concentration of Gardenia jasminoides Ellis extract alone for antibacterial is 50 mg / mL, and the minimum inhibitory concentration of Cinnamomum cassia Presl extract alone for antibacterial is 25 mg / mL. The antibacterial concentration required when the two are combined is much lower than that required for either extract alone, indicating that there is a good synergistic antibacterial effect between the two. The existing understanding of the antifungal mechanism mainly includes inhibiting the synthesis of sterols in the fungal cell membrane, inhibiting the synthesis of the fungal cell wall, and inhibiting the synthesis of nucleic acids in the fungus. However, there is no clear literature report on the antifungal mechanism of traditional Chinese medicine at present. Some studies have shown that cinnamon components can affect the permeability of the bacterial cell membrane. Combining the above test results, it is speculated that after the combination of the two extracts in the present invention affects the permeability of the Microsporum canis cell membrane to a certain extent, the Gardenia jasminoides Ellis extract enters the Microsporum canis cells and reaches a certain concentration to interfere with its cell synthesis, thus achieving a synergistic inhibitory effect on Microsporum canis.

[0032] Experiment on the treatment effect of clinical dermatophytosis in dogs and cats: Take 12 parts by weight of Gardenia jasminoides Ellis extract and 6 parts by weight of Cinnamomum cassia Presl extract, and use conventional methods and additives to make an ointment. This product is used to treat dogs and cats suffering from dermatophytosis. The selection criteria for dogs and cats are as follows: dogs and cats often scratch the affected area, the affected area is a round or oval hairless area with erythema, superficial scales and scabs at the edge. The affected area of the skin is irradiated with a Wood's lamp, and the affected area shows yellow-green fluorescence. Clinically, it is judged as dermatophytosis caused by Microsporum canis. The usage method is to apply an appropriate amount to the affected area several times a day. The treatment effect can be divided into three categories: cured, effective and ineffective. Cured means that after medication, the inflammatory secretions at the skin affected area completely disappear, there is no odor at the affected part, and symptoms such as red rashes, dandruff, and pustules all disappear. Effective means that after medication, the inflammatory secretions at the skin affected area are significantly reduced, there is almost no odor at the affected part, and symptoms such as red rashes, dandruff, and pustules are alleviated. Ineffective means that after medication, the inflammatory secretions at the skin affected area still exist, there is still an odor at the affected part, and symptoms such as red rashes, dandruff, and pustules have not improved. Apply the above ointment to 8 dogs with dermatophytosis. After 1 week of treatment, 6 were cured and 2 were effective. Continue to treat for 1 week, and all were cured. There was no recurrence after stopping the medication for 1 week. Apply the above ointment to 10 cats with dermatophytosis. After 1 week of treatment, all were cured. There was no recurrence after stopping the medication for 1 week. It can be seen that the product of the present invention can effectively treat dermatophytosis in dogs and cats, and has a low recurrence rate.

[0033] The above embodiments are only used to illustrate the technical concept and features of the present invention, and the purpose is to enable those who are familiar with this technology to understand the content of the present invention and implement it accordingly. It is not intended to limit the protection scope of the present invention. Any equivalent transformation or modification made according to the spirit and essence of the present invention should be covered within the protection scope of the present invention.

Claims

1. An antibacterial composition, characterized in that: The antibacterial composition is composed of 12-25 parts of gardenia extract and 6-12 parts of cinnamon extract in parts by mass.

2. The antibacterial composition according to claim 1, characterized in that: The gardenia extract is specifically obtained by crushing the gardenia, adding an appropriate volume of ethanol with a concentration of 75% (v / v) or more, ultrasonically extracting, filtering, and concentrating under reduced pressure.

3. The antibacterial composition according to claim 1, characterized in that: The cinnamon extract is specifically obtained by crushing cinnamon, adding an appropriate volume of ethanol with a concentration of 75% (v / v) or more, ultrasonically extracting, filtering, and concentrating under reduced pressure.

4. Use of the antibacterial composition according to any one of claims 1 to 3 in treating dermatophytosis in cats and dogs, characterized in that: The pathogenic bacteria of the cat and dog dermatophytosis is Microsporum canis.

5. An antibacterial composition ointment, characterized in that: The raw materials of the antibacterial composition ointment include the antibacterial composition according to any one of claims 1 to 4 and conventional excipients.

6. The method for preparing the antibacterial composition ointment according to any one of claims 4-5, characterized in that: Take 12-25 parts of gardenia extract and 6-12 parts of cinnamon extract, add conventional auxiliary materials, and adopt a conventional ointment preparation method to prepare an ointment.

7. Use of the antibacterial composition ointment according to claim 5 in treating dermatophytosis in cats and dogs, characterized in that: The pathogenic bacteria of the cat and dog dermatophytosis is Microsporum canis.

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