Chinese herbal medicine atomizing agent as well as preparation method and application thereof

Abstract

The invention belongs to the field of Chinese herbal medicine application, and particularly relates to a Chinese herbal medicine atomizing agent and a preparation method and application thereof. The Chinese herbal medicine atomizing agent comprises a plant extracting solution, antibacterial composite peptide and the like, the plant extracting solution is obtained by sequentially treating Chinese herbal medicines such as radix bupleuri, liquorice, blackberry lily, rheum officinale, honeysuckle and fructus forsythiae in three steps, and the Chinese herbal medicines have the effects of lifting yang qi, clearing away heat and toxic materials and relieving swollen sore throat; the antibacterial composite peptide is obtained by extracting a peptide component of andrographis paniculata and combining with curcumin modification, has broad-spectrum antibacterial activity and can inhibit the activity of pathogens related to amygdalitis. The Chinese herbal medicine atomizing agent disclosed by the invention is subjected to high-speed homogenization and ultrasonic treatment to form an oil-in-water microemulsion, and the form can better stabilize bioactive components in the Chinese herbal medicine atomizing agent. The obtained Chinese herbal medicine atomizing agent has a better anti-inflammatory effect, can greatly relieve symptoms such as throat pain, fever and dyspnea related to amygdalitis after being used, and has a better application prospect.

Description

Technical Field

[0001] The present invention belongs to the field of application of Chinese herbal medicines, and particularly relates to a Chinese herbal medicine atomizing agent, a preparation method thereof and an application thereof. Background Art

[0002] Tonsillitis is a common inflammatory disease of the lymphoid tissue mass at the root of the tongue, mainly divided into acute tonsillitis and chronic tonsillitis. Acute tonsillitis is usually caused by overwork or catching a cold, and chronic tonsillitis is caused by repeated attacks of acute tonsillitis. The symptoms of tonsillitis usually include fever, sore throat, dry throat, bad breath and foreign body sensation, snoring, etc. For adults, mild tonsillitis can heal on its own, while moderate to severe cases require timely medical treatment such as taking medicine. Pediatric tonsillitis is prone to occur in autumn and winter and is a common childhood disease. Since children's immune systems are not fully developed, they are easily invaded by pathogens.

[0003] Western medicines such as penicillin, amoxicillin, azithromycin, etc. are applicable to tonsillitis caused by bacterial infections. Hormonal drugs such as dexamethasone can be used to reduce the inflammatory response, relieve severe sore throat and swelling, and antipyretic and analgesic drugs such as paracetamol and ibuprofen can be used to relieve fever, etc. The classic traditional Chinese medicine formula selects Dabu Yin Pills combined with Wuwei Xiaodu Yin, which involves Chinese medicinal materials such as raw rehmannia root, tortoise shell, stir-fried phellodendron bark, anemarrhena rhizome, dandelion, viola yedoensis, honeysuckle flower, burdock fruit, platycodon root, liquorice root, etc., to achieve the effects of tonifying the deficient yin, purging the excessive fire, clearing the fire of the triple energizer, etc. Or select Danggui Liuhuang Decoction in "Secret Collection of the Orchid Chamber", which contains angelica sinensis, prepared rehmannia root, raw rehmannia root, scutellaria baicalensis, phellodendron bark, coptis chinensis, astragalus membranaceus, and has the effects of clearing heat and purging fire, replenishing qi and consolidating the exterior, nourishing yin and enriching blood, etc. Or select Modified Yupingfeng Powder, with astragalus membranaceus as the monarch drug, atractylodes macrocephala as the minister drug, and ledebouriella seseloides as the assistant drug, and appropriately add other Chinese medicinal materials, which can tonify the qi of the lung and spleen, enhance the external defense function, and reduce the invasion of exogenous pathogens. In addition to the above oral medications, traditional Chinese medicine also provides external treatment methods such as gargling, throat blowing, acupoint application, acupoint injection, atomizing inhalation, enema, etc., which have good clinical application effects.

[0004] Compared with other forms of drugs, an atomizing agent converts the drug into an aerosol form through an atomizing inhalation device (such as an air compression atomizer, an ultrasonic atomizer, etc.), and directly acts on the respiratory tract and lungs after inhalation. Its advantage is that the drug can act on the target organ at a higher concentration, without passing through the digestive system and blood circulation, and the onset time is faster than oral or injection methods. Therefore, the dosage of general atomizing agents is much lower than oral or intravenous administration, with less systemic absorption and thus fewer side effects, especially suitable for children, the elderly or patients with weak liver and kidney functions. Summary of the Invention

[0005] Based on the above background, the present invention provides a Chinese herbal medicine atomizing agent, which contains plant extracts, antibacterial composite peptides, etc. The plant extracts are obtained by performing three-step treatment on Chinese herbal medicines such as Bupleurum chinense, Glycyrrhiza uralensis, Belamcanda chinensis, Rheum palmatum, Lonicera japonica, and Forsythia suspensa in sequence, aggregating the effects of the above Chinese herbal medicines in lifting yang qi, clearing heat and detoxifying, and relieving sore throat; the antibacterial composite peptide is the peptide component extracted from Andrographis paniculata and obtained after being modified by combining curcumin, and has broad-spectrum antibacterial properties and can inhibit the activity of pathogens related to tonsillitis. The Chinese herbal medicine atomizing agent of the present invention forms an oil-in-water microemulsion after high-speed homogenization and ultrasonic treatment, and this form can better stabilize the bioactive components in the Chinese herbal medicine atomizing agent. The Chinese herbal medicine atomizing agent prepared by the present invention has better anti-inflammatory effects, and can greatly relieve symptoms such as sore throat, fever, and dyspnea related to tonsillitis after use, and has better application prospects.

[0006] The present invention provides a Chinese herbal medicine atomizing agent, which contains the following raw materials in parts by weight: 20-30 parts of plant extracts, 0.2-0.4 parts of vitamin D, and 1-3 parts of antibacterial composite peptides.

[0007] The preparation method of the plant extracts is as follows: L1. Grind and crush Bupleurum chinense and Glycyrrhiza uralensis, sieve through a 2-mesh sieve, weigh according to a mass ratio of 1:1-1.5 to obtain a mixed powder A, add water and stir until evenly dispersed to obtain a mixed solution, then add a compound microbial agent to the mixed solution, perform anaerobic fermentation at 37°C for 5-6 h, inactivate at high temperature after completion, filter the fermentation broth to obtain a filtrate, and concentrate the filtrate to 0.4-0.6 times the original volume to obtain a concentrated solution; L2. Grind and crush Belamcanda chinensis and Rheum palmatum, sieve through a 10-mesh sieve, weigh according to a mass ratio of 0.3-0.5:1 to obtain a mixed powder B, weigh the mixed powder B, choline chloride, citric acid, and the concentrated solution obtained in step L1 according to a mass-volume ratio of 10-15 g:1-2 g:1.4-2.8 g:800-1000 mL, perform ultrasonic treatment at a power of 200-240 W for 2-3 h, and obtain a treated solution after completion; L3. Weigh Lonicera japonica, Forsythia suspensa, and Mentha haplocalyx according to a mass ratio of 1:0.8-1:0.9-1.2 to obtain a mixture, weigh the mixture and the treated solution obtained in step L2 according to a mass-volume ratio of 4-6 g:600-700 mL, heat to 35-45°C, stir and treat at a rotation speed of 180-200 rpm for 1.5-2 h, naturally cool to room temperature after completion, remove solid residues by coarse filtration, and centrifuge at a centrifugal force of 8000-10000 g for 20-30 min, and obtain a supernatant after completion. The supernatant is the plant extracts.

[0008] Preferably, the amount of water added in step L1 is 150 - 200 times the mass of the mixed powder A. The compound microbial agent is prepared by mixing Lactobacillus animalis subsp. lactis and Bifidobacterium breve in a mass ratio of 1:1. Both Lactobacillus animalis subsp. lactis and Bifidobacterium breve can be purchased from the China General Microbiological Culture Collection Center, and their preservation numbers are CGMCC No. 1.15623 and CGMCC No. 1.213 respectively. The preferred bacterial concentration in the compound microbial agent is 1×10 4 -10 6 cfu / mL. The addition amount of the compound microbial agent is 4 - 6% by volume of the mixed solution.

[0009] The preparation method of the antibacterial composite peptide is as follows: S1. Grind and crush andrographis paniculata to obtain andrographis paniculata powder with a particle size of 10 - 15 μm. Weigh andrographis paniculata powder and water according to a mass ratio of 1 - 3 g: 30 - 40 mL, heat to 50 - 55 °C, stir at a speed of 100 - 150 rpm for 8 - 10 min. After completion, adjust the pH to 6.5, add papain, maintain the above temperature and speed, and enzymatically hydrolyze for 1 - 1.5 h. After completion, inactivate the enzyme at high temperature, naturally cool to room temperature, and centrifuge at a centrifugal force of 5000 - 6000 g for 20 - 30 min to obtain the supernatant; S2. Adjust the pH of the supernatant obtained in step S2 to 7.0, pass through a nanofiltration membrane column with a molecular weight cut-off of 200 - 250 Da, and measure the conductivity at the same time. Collect the concentrated peptide solution with a conductivity below 400 μS / cm. The concentrated peptide solution is washed 3 - 5 times with deionized water, and then the peptide solution with a molecular weight between 1 - 3 kDa is collected by an ultrafiltration membrane. After vacuum freeze-drying, peptide powder is obtained; S3. Weigh egg yolk lecithin, cholesterol, curcumin, methanol and chloroform according to a mass ratio of 60 - 70 mg: 10 - 15 mg: 6 - 8 mg: 8 - 10 mL: 4 - 6 mL. Mix the above substances, stir until evenly dispersed, and then heat to 35 - 40 °C until the solvent completely evaporates, and collect the film-like substance; S4. Dissolve the peptide powder obtained in step S2 in water to obtain a peptide solution with a mass percentage concentration of 8 - 10%. Add the film-like substance obtained in step S3 to the peptide solution, and continuously stir at a speed of 100 - 120 rpm at room temperature for 2 - 3 h. After completion, perform ultrasonic treatment at a power of 180 - 200 W for 10 - 15 min. The obtained solution is vacuum freeze-dried to obtain the antibacterial composite peptide.

[0010] Preferably, the papain used in step S1 is preferably a product with an enzyme activity of 15 - 20 U / mg, and the addition amount of papain is 0.08 - 0.1 mg / mL.

[0011] Preferably, the mass volume concentration of the film-like substance in the peptide solution in step S4 is 0.5 - 0.8 mg / mL.

[0012] The present invention also provides a preparation method of a Chinese herbal medicine atomizing agent, and the specific steps are as follows: V1. Add the antibacterial composite peptide into soybean oil, heat it to 80-90 °C, and stir it at a rotation speed of 200-250 rpm for 10-15 min to obtain component A; V2. Add vitamin D into the plant extract, and stir until it is evenly dispersed to obtain component B; V3. Mix the component A obtained in step V1 with the component B obtained in step V2, then homogenize it at a rotation speed of 8000-9000 rpm for 3-5 min, and then perform ultrasonic treatment at a power of 400-420 W for 1-2 min. After that, the Chinese herbal medicine atomizing agent is obtained.

[0013] Preferably, the dosage of soybean oil in step V1 is 4-5 times the weight part of the antibacterial composite peptide.

[0014] The present invention also provides an application of the Chinese herbal medicine atomizing agent, and the specific content is as follows: The Chinese herbal medicine atomizing agent obtained by the present invention is applicable to acute and chronic tonsillitis.

[0015] The beneficial effects of the present invention are as follows: The Chinese herbal medicine atomizing agent of the present invention comprises a plant extract, an antibacterial composite peptide, etc. The plant extract is obtained by performing three-step treatment on Chinese herbal medicines such as Bupleurum chinense. First, a composite microbial agent is used to ferment Bupleurum chinense and Glycyrrhiza uralensis, and the obtained fermentation broth provides the liquid component for the second-step treatment. Secondly, deep eutectic solvents are used to treat Belamcanda chinensis and Rheum palmatum, and combined with ultrasonic extraction to obtain a treated liquid. Finally, Lonicera japonica, Forsythia suspensa, and Mentha haplocalyx are enzymatically hydrolyzed to finally obtain the plant extract of the present invention. During the entire extraction process, Bupleurum chinense and Glycyrrhiza uralensis are extracted three times, Belamcanda chinensis and Rheum palmatum are extracted twice, and Lonicera japonica, etc. are extracted once. According to the pharmacopoeia records, Bupleurum chinense is pungent, bitter, slightly cold, and belongs to the liver, gallbladder, and lung meridians, and can disperse and clear heat, soothe the liver and relieve depression, and lift yang qi; Glycyrrhiza uralensis is sweet, flat, and belongs to the heart, lung, spleen, and stomach meridians, and can invigorate the spleen and replenish qi, clear heat and detoxify, and harmonize various medicines; Belamcanda chinensis is bitter, cold, and belongs to the lung meridian, and can clear heat and detoxify, and relieve sore throat; Rheum palmatum is bitter, cold, and belongs to the spleen, stomach, large intestine, liver, and pericardium meridians, and can purge and reduce accumulation, clear heat and purge fire; Lonicera japonica is sweet, cold, and belongs to the lung, heart, and stomach meridians, and can disperse wind-heat; Forsythia suspensa is bitter, slightly cold, and belongs to the lung, heart, and small intestine meridians, and can reduce swelling and dissipate binds; Mentha haplocalyx is pungent, cool, and belongs to the lung and liver meridians, and can clear the head and eyes, and benefit the throat. The above-mentioned various medicines are harmonized to obtain the plant extract, and the plant extract has good medicinal effects, can relieve inflammation, and improve symptoms such as sore throat, fever, etc. caused by tonsillitis. The antibacterial composite peptide of the present invention is a peptide component extracted from Andrographis paniculata. The peptide component involves many lipophilic amino acids, and the peptide component is further modified with curcumin to obtain the antibacterial composite peptide. The antibacterial composite peptide has broad-spectrum antibacterial properties and can inhibit the activity of pathogens related to tonsillitis. Finally, the antibacterial composite peptide is mixed with soybean oil to obtain Component A (oil component), and the plant extract is mixed with vitamin D to obtain Component B (water component). After high-speed homogenization and ultrasonic treatment, a Chinese herbal medicine atomizing agent in the form of an oil-in-water microemulsion is obtained, and this form can better stabilize the bioactive components in the Chinese herbal medicine atomizing agent.

[0016] The Chinese herbal medicine atomizing agent prepared by the present invention has a better anti-inflammatory effect. After use, it can greatly relieve symptoms such as throat pain, fever, and dyspnea related to tonsillitis, and has a better application prospect. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] The drawings described herein are used to provide a further understanding of the present application, and constitute a part of the present application. The illustrative embodiments of the present application and their descriptions are used to explain the present application, and do not constitute an improper limitation of the present application. In the drawings: Figure 1 are the experimental results of the swelling inhibition rate of different Chinese herbal medicine atomizing agents; Figure 2 are the content result diagrams of high-sensitivity C-reactive protein in the control group and the treatment group before and after treatment; Figure 3 are the content result diagrams of white blood cell count in the control group and the treatment group before and after treatment; Figure 4 It is a graph showing the content results of interleukin-2 in the control group and the treatment group before and after treatment. Specific implementation manners

[0018] In order to more clearly illustrate the overall concept of this application, the following will be described in detail by way of examples in combination with the accompanying drawings of the specification. In the following description, a large number of specific details are given to provide a more thorough understanding of the present invention. However, it is obvious to those skilled in the art that the present invention can be implemented without one or more of these details. In other examples, in order to avoid confusion with the present invention, some technical features well known in the art are not described.

[0019] Example 1: This example provides a preparation method of a plant extract, and the specific steps are as follows: L1. Grind and crush Bupleurum chinense and Glycyrrhiza uralensis, sieve through a 2-mesh sieve, weigh according to a mass ratio of 1:1 to obtain a mixed powder A, add water and stir until evenly dispersed. The added amount of water is 150 times the mass of the mixed powder A to obtain a mixed solution. Then, add a compound microbial preparation to the mixed solution. The compound microbial preparation is prepared from Bifidobacterium animalis subsp. lactis and Bifidobacterium breve according to a mass ratio of 1:1. Both Bifidobacterium animalis subsp. lactis and Bifidobacterium breve are purchased from the China General Microbiological Culture Collection Center, and their preservation numbers are CGMCC No.1.15623 and CGMCC No.1.213 respectively. The bacterial concentration in the compound microbial preparation is 1×10 4 cfu / mL. The added amount of the compound microbial preparation is 4% by volume of the mixed solution. Carry out anaerobic fermentation at 37°C for 5 h, inactivate at high temperature after completion, filter the fermentation broth to obtain a filtrate, and concentrate the filtrate to 0.4 times the original volume to obtain a concentrated solution; L2. Grind and crush Belamcanda chinensis and Rheum palmatum, sieve through a 10-mesh sieve, weigh according to a mass ratio of 0.3:1 to obtain a mixed powder B. Weigh the mixed powder B, choline chloride, citric acid, and the concentrated solution obtained in step L1 according to a mass-volume ratio of 10 g:1 g:1.4 g:800 mL, and perform ultrasonic treatment at a power of 200 W for 2 h to obtain a treated solution after completion; L3. Weigh Flos Lonicerae, Forsythia suspensa, and Mentha haplocalyx according to a mass ratio of 1:0.8:0.9 to obtain a mixture. Weigh the mixture and the treated solution obtained in step L2 according to a mass-volume ratio of 4 g:600 mL, heat to 35°C, stir and process at a rotation speed of 180 rpm for 1.5 h, naturally cool to room temperature after completion, remove solid residues by coarse filtration, and then centrifuge at a centrifugal force of 8000 g for 20 min. After completion, obtain the supernatant, and the supernatant is the plant extract.

[0020] Example 2: This example provides a preparation method of a plant extract, and the specific steps are as follows: L1. Grind and pulverize Bupleuri Radix and Glycyrrhizae Radix. After passing through a 2-mesh sieve, weigh them according to a mass ratio of 1:1.2 to obtain mixed powder A. Add water and stir until evenly dispersed. The added amount of water is 180 times the mass of mixed powder A to obtain a mixed solution. Then add the compound microbial preparation to the mixed solution. The compound microbial preparation is prepared from Bifidobacterium animalis subsp. lactis and Bifidobacterium breve according to a mass ratio of 1:1. Both Bifidobacterium animalis subsp. lactis and Bifidobacterium breve are purchased from the China General Microbiological Culture Collection Center, and their preservation numbers are CGMCC No.1.15623 and CGMCC No.1.213 respectively. The bacterial concentration in the compound microbial preparation is 1×10 5 cfu / mL. The added amount of the compound microbial preparation is 5% by volume of the mixed solution. Carry out anaerobic fermentation at 37°C for 5.4 h. After completion, inactivate at high temperature. Filter the fermentation broth to obtain a filtrate, and concentrate the filtrate to 0.5 times the original volume to obtain a concentrated solution; L2. Grind and pulverize Belamcandae Rhizoma and Rhei Radix et Rhizoma. After passing through a 10-mesh sieve, weigh them according to a mass ratio of 0.4:1 to obtain mixed powder B. Weigh mixed powder B, choline chloride, citric acid, and the concentrated solution obtained in step L1 according to a mass-volume ratio of 12 g:1.5 g:2.1 g:900 mL, and perform ultrasonic treatment at a power of 220 W for 2.5 h. After completion, obtain a treated solution; L3. Weigh Flos Lonicerae, Fructus Forsythiae, and Herba Menthae according to a mass ratio of 1:0.9:1 to obtain a mixture. Weigh the mixture and the treated solution obtained in step L2 according to a mass-volume ratio of 5 g:650 mL, heat to 39°C, and stir and treat at a rotation speed of 190 rpm for 1.8 h. After completion, naturally cool to room temperature. After rough filtering to remove solid residues, centrifuge at a centrifugal force of 9000 g for 25 min. After completion, obtain the supernatant, and the supernatant is the plant extract.

[0021] Example 3: This example provides a method for preparing a plant extract, and the specific steps are as follows: L1. Grind and pulverize Bupleuri Radix and Glycyrrhizae Radix. After passing through a 2-mesh sieve, weigh them according to a mass ratio of 1:1.5 to obtain mixed powder A. Add water and stir until evenly dispersed. The added amount of water is 200 times the mass of mixed powder A to obtain a mixed solution. Then add the compound microbial preparation to the mixed solution. The compound microbial preparation is prepared from Bifidobacterium animalis subsp. lactis and Bifidobacterium breve according to a mass ratio of 1:1. Both Bifidobacterium animalis subsp. lactis and Bifidobacterium breve are purchased from the China General Microbiological Culture Collection Center, and their preservation numbers are CGMCC No.1.15623 and CGMCC No.1.213 respectively. The bacterial concentration in the compound microbial preparation is 1×10 6cfu / mL. The addition amount of the compound microbial agent is 6% by volume of the mixed solution. Anaerobic fermentation is carried out at 37 °C for 6 h. After completion, high-temperature inactivation is carried out. The fermentation broth is filtered to obtain a filtrate, and the filtrate is concentrated to 0.6 times the original volume to obtain a concentrated solution. L2. Belamcanda chinensis and Rheum palmatum are ground and pulverized, and after passing through a 10-mesh sieve, they are weighed according to a mass ratio of 0.5:1 to obtain a mixed powder B. The mixed powder B, choline chloride, citric acid, and the concentrated solution obtained in step L1 are weighed according to a mass-volume ratio of 15 g:2 g:2.8 g:1000 mL, and ultrasonic treatment is carried out at a power of 240 W for 3 h. After completion, a treated solution is obtained. L3. Flos Lonicerae, Fructus Forsythiae, and Mentha haplocalyx are weighed according to a mass ratio of 1:1:1.2 to obtain a mixture. The mixture and the treated solution obtained in step L2 are weighed according to a mass-volume ratio of 6 g:700 mL, heated to 45 °C, and stirred at a rotation speed of 200 rpm for 2 h. After completion, it is naturally cooled to room temperature. After rough filtration to remove solid residues, it is centrifuged at a centrifugal force of 10000 g for 30 min. After completion, the supernatant is obtained, and the supernatant is the plant extract.

[0022] Example 4: This example provides a preparation method of an antibacterial composite peptide, and the specific steps are as follows: S1. Andrographis paniculata is ground and pulverized to obtain andrographis paniculata powder with a particle size of 10 μm. The andrographis paniculata powder and water are weighed according to a mass ratio of 1 g:30 mL, heated to 50 °C, and stirred at a rotation speed of 100 rpm for 8 min. After completion, the pH is adjusted to 6.5, and papain is added. The papain is a product with an enzyme activity of 15 U / mg, and the addition amount of papain is 0.08 mg / mL. Keeping the above temperature and rotation speed, enzymatic hydrolysis is carried out for 1 h. After completion, the enzyme is inactivated at high temperature, and it is naturally cooled to room temperature and centrifuged at a centrifugal force of 5000 g for 20 min to obtain a supernatant. S2. The pH of the supernatant obtained in step S2 is adjusted to 7.0, and it is passed through a 200 Da nanofiltration membrane column. At the same time, the conductivity is measured, and the concentrated peptide solution with a conductivity below 400 μS / cm is collected. The concentrated peptide solution is washed 3 times with deionized water, and then the peptide solution between 1 - 3 kDa is collected by an ultrafiltration membrane and vacuum freeze-dried to obtain a peptide powder. S3. Egg yolk lecithin, cholesterol, curcumin, methanol, and chloroform are weighed according to a mass ratio of 60 mg:10 mg:6 mg:8 mL:4 mL. The above substances are mixed and stirred until evenly dispersed, and then heated to 35 °C until the solvent is completely volatilized, and the film-like substance is collected. S4. Dissolve the peptide powder obtained in step S2 in water to obtain a peptide solution with a mass percentage concentration of 8%. Add the film-like substance obtained in step S3 to the peptide solution. The mass-volume concentration of the film-like substance in the peptide solution is 0.5 mg / mL. Continuously stir at a speed of 100 rpm at room temperature for 2 h. After that, perform ultrasonic treatment at a power of 180 W for 10 min. Vacuum freeze-dry the obtained solution to obtain the antibacterial composite peptide.

[0023] Example 5: This example provides a method for preparing an antibacterial composite peptide, and the specific steps are as follows: S1. Grind and crush andrographis paniculata to obtain andrographis paniculata powder with a particle size of 12 μm. Weigh andrographis paniculata powder and water according to a mass ratio of 2 g:35 mL, heat to 52 °C, stir and process at a speed of 120 rpm for 9 min. After that, adjust the pH to 6.5, add papain, which is a product with an enzyme activity of 18 U / mg. The addition amount of papain is 0.09 mg / mL. Maintain the above temperature and speed and enzymatically hydrolyze for 1.2 h. After that, inactivate the enzyme at high temperature and naturally cool to room temperature. Centrifuge at a centrifugal force of 5400 g for 25 min to obtain the supernatant. S2. Adjust the pH of the supernatant obtained in step S2 to 7.0, pass through a 220 Da nanofiltration membrane column, and measure the conductivity at the same time. Collect the concentrated peptide solution with a conductivity below 400 μS / cm. Wash the concentrated peptide solution 4 times with deionized water, and then collect the peptide solution between 1 - 3 kDa by an ultrafiltration membrane. Vacuum freeze-dry to obtain the peptide powder. S3. Weigh egg yolk lecithin, cholesterol, curcumin, methanol and chloroform according to a mass ratio of 65 mg:12 mg:7 mg:9 mL:5 mL. Mix the above substances, stir until evenly dispersed, and heat to 38 °C until the solvent completely evaporates. Collect the film-like substance. S4. Dissolve the peptide powder obtained in step S2 in water to obtain a peptide solution with a mass percentage concentration of 9%. Add the film-like substance obtained in step S3 to the peptide solution. The mass-volume concentration of the film-like substance in the peptide solution is 0.7 mg / mL. Continuously stir at a speed of 110 rpm at room temperature for 2.5 h. After that, perform ultrasonic treatment at a power of 190 W for 12 min. Vacuum freeze-dry the obtained solution to obtain the antibacterial composite peptide.

[0024] Example 6: This example provides a method for preparing an antibacterial composite peptide, and the specific steps are as follows: S1. Grind and powder Andrographis paniculata to obtain Andrographis paniculata powder with a particle size of 15 μm. Weigh Andrographis paniculata powder and water according to a mass ratio of 3 g:40 mL, heat to 55 °C, stir at a speed of 150 rpm for 10 min. After that, adjust the pH to 6.5, add papain, which is a product with an enzyme activity of 20 U / mg, and the addition amount of papain is 0.1 mg / mL. Maintain the above temperature and speed, and enzymatically hydrolyze for 1.5 h. After completion, inactivate the enzyme at high temperature, naturally cool to room temperature, and centrifuge at a centrifugal force of 6000 g for 30 min to obtain the supernatant; S2. Adjust the pH of the supernatant obtained in step S2 to 7.0, pass through a 250 Da nanofiltration membrane column, and measure the conductivity at the same time. Collect the concentrated peptide solution with a conductivity below 400 μS / cm. Wash the concentrated peptide solution 5 times with deionized water, and then collect the peptide solution between 1 - 3 kDa using an ultrafiltration membrane. After vacuum freeze-drying, obtain the peptide powder; S3. Weigh egg yolk lecithin, cholesterol, curcumin, methanol, and chloroform according to a mass ratio of 70 mg:15 mg:8 mg:10 mL:6 mL. Mix the above substances, stir until evenly dispersed, and then heat to 40 °C until the solvent completely evaporates, and collect the film-like substance; S4. Dissolve the peptide powder obtained in step S2 in water to obtain a peptide solution with a mass percentage concentration of 10%. Add the film-like substance obtained in step S3 to the peptide solution, and the mass volume concentration of the film-like substance in the peptide solution is 0.8 mg / mL. Continuously stir at a speed of 120 rpm at room temperature for 3 h. After completion, perform ultrasonic treatment at a power of 200 W for 15 min. The obtained solution is vacuum freeze-dried to obtain the antibacterial composite peptide.

[0025] Example 7: This example provides a preparation method of a Chinese herbal medicine atomizing agent, and the specific steps are as follows: The Chinese herbal medicine atomizing agent in this example contains the following raw materials in parts by weight: 20 parts of plant extract, 0.2 part of vitamin D, 1 part of antibacterial composite peptide, wherein the plant extract is prepared in Example 1, and the antibacterial composite peptide is prepared in Example 4; V1. Add the antibacterial composite peptide to soybean oil, and the amount of soybean oil used is 4 times the weight part of the antibacterial composite peptide. Heat to 80 °C and stir at a speed of 200 rpm for 10 min to obtain component A; V2. Add vitamin D to the plant extract and stir until evenly dispersed to obtain component B; V3. Mix the component A obtained in step V1 and the component B obtained in step V2, then homogenize at a speed of 8000 rpm for 3 min, and then perform ultrasonic treatment at a power of 400 W for 1 min. After completion, the Chinese herbal medicine atomizing agent is obtained.

[0026] Example 8: This example provides a preparation method of a Chinese herbal medicine atomizing agent, and the specific steps are as follows: The Chinese herbal medicine atomizing agent of this example contains the following raw materials in parts by weight: 24 parts of plant extract, 0.3 part of vitamin D, and 2 parts of antibacterial composite peptide. The plant extract is obtained by the preparation in Example 2, and the antibacterial composite peptide is obtained by the preparation in Example 5; V1. Add the antibacterial composite peptide to soybean oil. The amount of soybean oil used is 4.5 times the weight part of the antibacterial composite peptide. Heat it to 85 °C and stir it at a speed of 220 rpm for 12 min to obtain Component A; V2. Add vitamin D to the plant extract and stir until evenly dispersed to obtain Component B; V3. Mix the Component A obtained in step V1 with the Component B obtained in step V2, then homogenize it at a speed of 8500 rpm for 4 min, and then perform ultrasonic treatment at a power of 410 W for 1.5 min. After that, the Chinese herbal medicine atomizing agent is obtained.

[0027] Example 9: This example provides a preparation method of a Chinese herbal medicine atomizing agent, and the specific steps are as follows: The Chinese herbal medicine atomizing agent of this example contains the following raw materials in parts by weight: 30 parts of plant extract, 0.4 part of vitamin D, and 3 parts of antibacterial composite peptide. The plant extract is obtained by the preparation in Example 3, and the antibacterial composite peptide is obtained by the preparation in Example 6; V1. Add the antibacterial composite peptide to soybean oil. The amount of soybean oil used is 5 times the weight part of the antibacterial composite peptide. Heat it to 90 °C and stir it at a speed of 250 rpm for 15 min to obtain Component A; V2. Add vitamin D to the plant extract and stir until evenly dispersed to obtain Component B; V3. Mix the Component A obtained in step V1 with the Component B obtained in step V2, then homogenize it at a speed of 9000 rpm for 5 min, and then perform ultrasonic treatment at a power of 420 W for 2 min. After that, the Chinese herbal medicine atomizing agent is obtained.

[0028] Comparative Example 1: This comparative example provides a preparation method of a Chinese herbal medicine atomizing agent, and the specific steps are as follows: The difference between this comparative example and Example 7 is that it does not contain plant extract, and other raw materials and steps are the same as those in Example 7, that is, the Chinese herbal medicine atomizing agent of this comparative example is obtained.

[0029] Comparative Example 2: This comparative example provides a preparation method of a Chinese herbal medicine atomizing agent, and the specific steps are as follows: The difference between this comparative example and Example 8 is that it does not contain antibacterial composite peptide and contains soybean oil, and other raw materials and steps are the same as those in Example 8, that is, the Chinese herbal medicine atomizing agent of this comparative example is obtained.

[0030] Comparative Example 3: This comparative example provides a method for preparing a Chinese herbal medicine atomizing agent, and the specific steps are as follows: The difference between this comparative example and Example 9 is that it does not involve soybean oil, and other raw materials and steps are the same as those in Example 9, that is, the Chinese herbal medicine atomizing agent of this comparative example is obtained.

[0031] Experimental test: I. Animal experiment: Male Kunming mice with a body weight of 20±2 g were randomly divided into 5 groups, with 10 mice in each group, namely the blank control group, the positive aspirin group, the high-dose group of the Chinese herbal medicine atomizing agent, the medium-dose group of the Chinese herbal medicine atomizing agent, and the low-dose group of the Chinese herbal medicine atomizing agent. Aspirin (Guangdong Bedi Pharmaceutical Co., Ltd., approval number H44024478) was administered by gavage at a dose of 20 mL / kg once a day for one week. The blank control group was given an equal volume of distilled water. The Chinese herbal medicine atomizing agent was administered by a handheld animal lung spray dosing system after atomization treatment. The high-dose group was at a dose of 2 mL / kg, the medium-dose group was at a dose of 1 mL / kg, and the low-dose group was at a dose of 0.5 mL / kg, once a day for one week. 45 minutes after the last dose, 20 μL of xylene was dropped on both sides of the right ear of each mouse to cause inflammation. 15 minutes after inflammation induction, the mice were sacrificed by cervical dislocation. The two ears were cut off at the ear root, and ear pieces of equal area were punched at the same position on both ears with a 6 mm punch. The ear pieces were weighed with an electronic balance, and the difference between the right ear and the left ear was used as the swelling degree to calculate the ear piece weight coefficient and the swelling inhibition rate: Swelling inhibition rate = (swelling degree of the control group - swelling degree of the experimental group) / swelling degree of the control group × 100%.

[0032] The experimental results are as Figure 1 shown. It can be seen from Figure 1 that aspirin can effectively relieve the inflammation caused by xylene. The Chinese herbal medicine atomizing agents prepared in Examples 7-9 also have good anti-inflammatory effects, while Comparative Examples 1-3 with incomplete raw material formulas have poor effects. This shows that there is a synergistic interaction between the plant extract and the antibacterial composite peptide. At the same time, the atomizing agent of the present invention is prepared into an oil-in-water emulsion, which helps to maintain the stability of the use effect of the atomizing agent.

[0033] Volunteer experiment: 60 volunteers were recruited, aged between 24 and 52 years old. Symptoms: acute onset, burning pain in the throat, increased pain on swallowing, fever and aversion to cold, general discomfort and other self-perceived symptoms; Examination: tonsils were swollen and congested, and in severe cases, the crypt openings were yellowish-white, with pus beans or pseudomembranes formed, but not exceeding the tonsils themselves, and were easily wiped off without bleeding. The pharynx was congested, and the submandibular lymph nodes were tender and swollen; Laboratory examination: the total number of peripheral blood white blood cells > 10×10 9 / L, with a neutral increase. Patients meeting the above conditions suffered from acute tonsillitis and were randomly divided into two groups, namely the control group and the treatment group. In the control group, dexamethasone tablets (Guangdong Nanguo Pharmaceutical Co., Ltd., approval number H44024618) at 5 mg were diluted with 20 mL of 0.9% normal saline and atomized and inhaled through an atomizer. In the treatment group, 4 mL of the Chinese herbal medicine atomizing agent prepared in Example 9 was used. Both the control group and the treatment group were administered 3 times a day. The dosage was reduced after the symptoms were relieved, and the treatment effect was evaluated after 7 days.

[0034] Before and after treatment, an enzyme-linked immunosorbent assay kit was used to detect the contents of hypersensitive C-reactive protein, white blood cell count, and interleukin-2, and the inflammatory reaction was observed. At the same time, the total effective rate of treatment was calculated. The total effective rate of treatment = (number of cured cases + number of markedly effective cases + number of improved cases) / total number of cases × 100%. Cured: The clinical related symptoms of the patient basically disappeared, and the inflammatory factors returned to normal levels, and the patient could eat and drink normally; Markedly effective: The clinical related symptoms of the patient were significantly improved, the tonsils basically returned to the normal state, there was slight sore throat, and the inflammatory factors decreased significantly compared with before treatment, and the patient could eat a proper amount of liquid food; Improved: The clinical related symptoms of the patient were somewhat relieved, the sore throat was reduced compared with before treatment, but there were still symptoms of swelling and congestion; Ineffective: The related symptoms of the patient persisted, the sore throat was obvious, the patient could not eat, and there were symptoms such as high fever and dyspnea.

[0035] The inflammatory reaction conditions before and after treatment in the control group and the treatment group are respectively as Figure 2 , Figure 3 , Figure 4 shown. It can be seen from Figures 2 - 4 that the Chinese herbal medicine atomizing agent prepared in Example 9 has a good application effect and is better than dexamethasone aerosol in relieving acute tonsillitis. The treatment effects of the control group and the treatment group are shown in Table 1. It can be seen from Table 1 that after 7 days of treatment, the number of cured and markedly effective cases in the treatment group of volunteers is higher than that in the control group, and the total effective rate is also significantly higher than that in the control group, proving that the Chinese herbal medicine atomizing agent obtained in Example 9 has a better application prospect.

[0036] Table 1 Comparison of treatment effects

[0037] As mentioned above, these are only several embodiments of the present invention and do not impose any form of limitation on the present invention. Although the present invention is disclosed as above with preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art, without departing from the scope of the technical solution of the present invention, making some changes or modifications using the disclosed technical content is equivalent to equivalent implementation cases and all fall within the scope of protection of the technical solution of the present invention.

Claims

1. A Chinese herbal medicine atomizing agent, characterized in that, Comprising the following raw materials in parts by weight: 20 - 30 parts of plant extract, 0.2 - 0.4 parts of vitamin D, 1 - 3 parts of antibacterial composite peptide.

2. The Chinese herbal medicine atomizing agent according to claim 1, characterized in that, The preparation method of the plant extract is as follows: L1. Grind and pulverize bupleurum and licorice, weigh them according to the mass ratio of 1:1 - 1.5 to obtain mixed powder A, add water and stir until evenly dispersed to obtain a mixed solution, then add the composite microbial agent to the mixed solution for anaerobic fermentation. After completion, inactivate at high temperature. After filtering the fermentation broth, obtain a filtrate, and concentrate the filtrate to 0.4 - 0.6 times the original volume to obtain a concentrated solution; L2. Grind and pulverize belamcanda chinensis and rhubarb, weigh them according to the mass ratio of 0.3 - 0.5:1 to obtain mixed powder B. Weigh mixed powder B, choline chloride, citric acid, and the concentrated solution obtained in step L1 according to the mass - volume ratio of 10 - 15 g:1 - 2 g:1.4 - 2.8 g:800 - 1000 mL, and perform ultrasonic treatment. After completion, obtain a treated solution; L3. Weigh honeysuckle, forsythia, and mint according to the mass ratio of 1:0.8 - 1:0.9 - 1.2 to obtain a mixture. Weigh the mixture and the treated solution obtained in step L2 according to the mass - volume ratio of 4 - 6 g:600 - 700 mL, heat and stir - process. After completion, naturally cool to room temperature. After rough - filtering to remove solid residues, centrifuge. After completion, obtain the supernatant, and the supernatant is the plant extract.

3. The Chinese herbal medicine atomizing agent according to claim 2, wherein In step L1, the addition amount of water is 150 - 200 times the mass of mixed powder A. The composite microbial agent is prepared from Lactobacillus bifidus animalis subsp. lactis and Bifidobacterium breve according to the mass ratio of 1:

1. The addition amount of the composite microbial agent is 4 - 6% of the mixed solution by volume.

4. The Chinese herbal medicine atomizing agent according to claim 3, characterized in that, The preparation method of the antibacterial composite peptide is as follows: S1. Grind and pulverize andrographis paniculata to obtain andrographis paniculata powder. Weigh andrographis paniculata powder and water according to the mass ratio of 1 - 3 g:30 - 40 mL, heat and stir - process, add papain for enzymatic hydrolysis. After completion, inactivate the enzyme at high temperature, naturally cool to room temperature, and centrifuge to obtain the supernatant; S2. Adjust the pH of the supernatant obtained in step S2 to 7.0, pass through a nanofiltration membrane column with a molecular weight cut - off of 200 - 250 Da, and measure the conductivity simultaneously. Collect the concentrated peptide solution with a conductivity below 400 μS / cm. Wash the concentrated peptide solution with deionized water, and then collect the peptide solution with a molecular weight between 1 - 3 kDa using an ultrafiltration membrane. After vacuum freeze - drying, obtain peptide powder; S3. Weigh egg yolk lecithin, cholesterol, curcumin, methanol, and chloroform according to the mass ratio of 60 - 70 mg:10 - 15 mg:6 - 8 mg:8 - 10 mL:4 - 6 mL. Mix the above substances, stir until evenly dispersed, and then heat until the solvent completely evaporates, and collect the film - like substance; S4. Dissolve the peptide powder obtained in step S2 in water to obtain a peptide solution with a mass - percentage concentration of 8 - 10%. Add the film - like substance obtained in step S3 to the peptide solution, continuously stir at room temperature. After completion, perform ultrasonic treatment. The obtained solution is vacuum freeze - dried to obtain the antibacterial composite peptide.

5. The Chinese herbal medicine atomizing agent according to claim 4, wherein In step S1, the addition amount of papain is 0.08 - 0.1 mg / mL.

6. The Chinese herbal medicine atomizing agent according to claim 5, wherein In step S4, the mass - volume concentration of the film - like substance in the peptide solution is 0.5 - 0.8 mg / mL.

7. A preparation method of the Chinese herbal medicine atomizing agent according to any one of claims 1-6, characterized in that, The specific steps are as follows: V1. Add the antibacterial composite peptide into soybean oil, heat and stir to obtain component A; V2. Add vitamin D into the plant extract and stir until evenly dispersed to obtain component B; V3. Mix the component A obtained in step V1 with the component B obtained in step V2, homogenize and perform ultrasonic treatment, and then the Chinese herbal medicine atomizing agent is obtained after completion.

8. The preparation method of the Chinese herbal medicine atomizing agent according to claim 7, characterized in that, In step V1, the dosage of soybean oil is 4-5 times the weight part of the antibacterial composite peptide.

9. Use of the Chinese herbal medicine atomizing agent according to any one of claims 1-6, characterized in that, The Chinese herbal medicine atomizing agent obtained by the present invention is applicable to acute and chronic tonsillitis.

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