Antibiotic-loaded nano-emulsion as well as preparation method and application thereof

Abstract

The invention provides an antibiotic-loaded nano-emulsion and a preparation method and application thereof, and relates to the technical field of medical excipients, antibiotics are mixed with pharmaceutical excipients including soybean oil, dipalmitoyl phosphatidylcholine and cholesterol, and the mixture is dissolved by a volatile organic solvent to form an oil phase; sodium deoxycholate and Tween are added into the sterile liquid to be dissolved to form a water phase; and mixing the oil phase and the water phase, and carrying out ultrasonic treatment, rotary evaporation treatment and high-pressure homogenization treatment to obtain the antibiotic-loaded nano-emulsion. By pertinently adjusting and matching various factors such as types of inhalable pharmaceutic adjuvants, the adding proportion of the inhalable pharmaceutic adjuvants, the ratio of the oil phase to the water phase, the ultrasonic power, the homogenizing pressure and the like, the prepared nano-emulsion can efficiently load antibiotics, has good aerosol inhalation performance and better drug stability, and can be used for preparing the antibiotic-loaded nano-emulsion. The nanoemulsion can remain in lung tissues of mice for a long time, and the delivery efficiency of lung drugs is remarkably improved.

Description

Technical Field

[0001] The present application relates to the technical field of medical excipients, and in particular to a nanoemulsion loaded with antibiotics, and a preparation method and application thereof. Background Art

[0002] At present, there is a serious shortage of inhaled antibiotics for clinical use. The only inhaled antibiotic approved for clinical use in China is tobramycin inhalation solution, and many antibiotics such as rifampicin and polymyxin have been used for inhalation beyond the instructions. Therefore, it is urgent to develop antibiotic preparations specifically for inhalation.

[0003] The existing inhaled antibiotics are mainly solution-type inhaled antibiotics, and the only nano-preparation for inhalation on the market is amikacin inhaled liposome suspension. However, solution-type inhaled antibiotics have the disadvantages of rapid blood entry, low lung retention, large dosage, and insufficient antibacterial efficiency.

[0004] Based on this, a new antibiotic-loaded nanoformulation for inhalation is urgently needed. Summary of the invention

[0005] The present application provides an antibiotic-loaded nanoemulsion and a preparation method and application thereof, so as to solve the problems of existing inhaled antibiotics such as rapid blood entry, low lung retention, large dosage, and insufficient antibacterial efficiency.

[0006] In a first aspect of the embodiments of the present application, a method for preparing an antibiotic-loaded nanoemulsion is provided, the method comprising the following steps: Step 1: Evenly mix antibiotics, soybean oil, dipalmitoylphosphatidylcholine and cholesterol to obtain a mixture, and dissolve the mixture in a volatile organic solvent to prepare an oil phase, wherein 0.1 to 20 mg of antibiotics, 0.1 to 100 mg of soybean oil, 1 to 100 mg of dipalmitoylphosphatidylcholine and 0.1 to 3 mg of cholesterol are added to each 1 ml of the volatile organic solvent in the oil phase; Step 2: adding an organic acid sodium salt and Tween to a sterile liquid, stirring until the organic acid sodium salt and the Tween are dissolved, to prepare an aqueous phase, wherein 0.1 to 4 mg of the organic acid sodium salt and 0.1 to 5 mg of Tween are added to each 1 ml of the sterile liquid in the aqueous phase; Step 3: After the oil phase and the water phase are uniformly mixed, ultrasonic treatment is performed to obtain a first emulsion, and the volatile organic solvent in the first emulsion is removed by rotary evaporation to obtain a second emulsion, wherein the mixing volume ratio of the volatile organic solvent in the oil phase to the sterile liquid in the water phase is 1 to 8:1; Step 4: subjecting the second emulsion to high pressure homogenization to obtain an antibiotic-loaded nanoemulsion.

[0007] Optionally, in step 1, the volatile organic solvent is at least one of dichloromethane, ethyl acetate, ether and chloroform.

[0008] Optionally, in step 1, the antibiotic is at least one of rifampicin, polymyxin and vancomycin.

[0009] Optionally, in step 1, the sterile liquid is at least one of water, physiological saline, deionized water, phosphate buffer, deoxycholic acid, citrate buffer and Tween aqueous solution.

[0010] Optionally, in step 2, the organic acid sodium salt is at least one of sodium citrate and sodium deoxycholate.

[0011] Optionally, in step 3, the power of the ultrasonic treatment is 30 watts to 400 watts, and the time of the ultrasonic treatment is 20 seconds to 20 minutes.

[0012] Optionally, in step 3, the temperature of the rotary evaporation to remove the volatile organic solvent in the first emulsion is 37 degrees Celsius to 60 degrees Celsius, and the time of the rotary evaporation to remove the volatile organic solvent in the first emulsion is 5 minutes to 10 minutes.

[0013] Optionally, in step 4, the pressure of the high-pressure homogenization treatment is 200 bar to 1000 bar, and the time of the high-pressure homogenization treatment is 2 minutes to 20 minutes.

[0014] In a second aspect of the embodiments of the present application, there is provided an antibiotic-loaded nanoemulsion, wherein the antibiotic-loaded nanoemulsion is prepared by the preparation method described in any one of the first aspects above.

[0015] In a third aspect of the embodiments of the present application, there is provided a use of the antibiotic-loaded nanoemulsion as described in the second aspect in the preparation of an inhaled antibiotic drug.

[0016] The present application has the following advantages: The present application proposes a nanoemulsion loaded with antibiotics and a preparation method and application thereof, the method comprising the following steps: uniformly mixing antibiotics, soybean oil, dipalmitoylphosphatidylcholine and cholesterol to obtain a mixed material, dissolving the mixed material with a volatile organic solvent to prepare an oil phase, in which 0.1 to 20 mg of antibiotics, 0.1 to 100 mg of soybean oil, 1 to 100 mg of dipalmitoylphosphatidylcholine and 0.1 to 3 mg of cholesterol are added to each milliliter of the volatile organic solvent; sodium salt of an organic acid and Tween are added to the mixture; Stirring in a sterile liquid until the organic acid sodium salt and the Tween are dissolved to prepare an aqueous phase, in which 0.1 to 4 mg of the organic acid sodium salt and 0.1 to 5 mg of Tween are added to every 1 ml of the sterile liquid; after the oil phase and the aqueous phase are evenly mixed, ultrasonic treatment is performed to obtain a first emulsion, and the volatile organic solvent in the first emulsion is removed by rotary evaporation to obtain a second emulsion, wherein the mixing volume ratio of the volatile organic solvent in the oil phase to the sterile liquid in the aqueous phase is 1 to 8:1; the second emulsion is subjected to high-pressure homogenization treatment to obtain an antibiotic-loaded nanoemulsion.

[0017] The present application specifically adjusts the types of inhalable pharmaceutical excipients, the addition ratio of inhalable pharmaceutical excipients, the ratio of the oil phase and the water phase, and various factors such as ultrasonic power and homogenization pressure, and cooperates with each other to enable the prepared nanoemulsion to efficiently load antibiotics and have good atomization inhalation performance and good drug stability. Compared with the inhaled antibiotic solution dosage form, the nanoemulsion can be retained in the lung tissue of mice for a longer time, significantly improving the efficiency of lung drug delivery. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings required for use in the description of the embodiments of the present application will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present application. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative labor.

[0019] Figure 1 This is a transmission electron microscopy morphology image of a rifampicin-loaded nanoemulsion provided in an embodiment of the present application; Figure 2 This is a transmission electron microscopy morphology of a nanoemulsion loaded with polymyxin provided in an embodiment of the present application; Figure 3 This is a transmission electron microscopy morphology image of a nanoemulsion loaded with vancomycin provided in an example of the present application; Figure 4 The example of the present application provides a quantitative detection diagram of lung tissue of a rifampicin nanoemulsion 48 hours after inhalation by mice. DETAILED DESCRIPTION

[0020] The following will be combined with the drawings in the embodiments of the present application to clearly and completely describe the technical solutions in the embodiments of the present application. Obviously, the described embodiments are part of the embodiments of the present application, not all of the embodiments. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of this application.

[0021] At present, there is a serious shortage of inhaled antibiotics for clinical use. The only inhaled antibiotic approved for clinical use in China is tobramycin inhalation solution, and many antibiotics such as rifampicin and polymyxin have been used for inhalation beyond the instructions. Therefore, it is urgent to develop antibiotic preparations specifically for inhalation.

[0022] The existing inhaled antibiotics are mainly solution-type inhaled antibiotics, and the only nano-preparation for inhalation on the market is amikacin inhaled liposome suspension. However, solution-type inhaled antibiotics have the disadvantages of rapid blood entry, low lung retention, large dosage, and insufficient antibacterial efficiency.

[0023] Based on this, the present application proposes a method for preparing a nanoemulsion loaded with antibiotics, using various inhalable pharmaceutical excipients, such as soybean oil, dipalmitoylphosphatidylcholine (DPPC), cholesterol, phospholipids and other materials to prepare a nanoemulsion that can load antibiotics.

[0024] In a first aspect of the present application, a method for preparing an antibiotic-loaded nanoemulsion is provided, the method comprising the following steps: Step 1: Evenly mix antibiotics, soybean oil, dipalmitoylphosphatidylcholine and cholesterol to obtain a mixture, and dissolve the mixture in a volatile organic solvent to prepare an oil phase, wherein 0.1 to 20 mg of antibiotics, 0.1 to 100 mg of soybean oil, 1 to 100 mg of dipalmitoylphosphatidylcholine and 0.1 to 3 mg of cholesterol are added to each 1 ml of the volatile organic solvent in the oil phase; Step 2: adding an organic acid sodium salt and Tween to a sterile liquid, stirring until the organic acid sodium salt and the Tween are dissolved, to prepare an aqueous phase, wherein 0.1 to 4 mg of the organic acid sodium salt and 0.1 to 5 mg of Tween are added to each 1 ml of the sterile liquid in the aqueous phase; Step 3: After the oil phase and the water phase are uniformly mixed, ultrasonic treatment is performed to obtain a first emulsion, and the volatile organic solvent in the first emulsion is removed by rotary evaporation to obtain a second emulsion, wherein the mixing volume ratio of the volatile organic solvent in the oil phase to the sterile liquid in the water phase is 1 to 8:1; Step 4: subjecting the second emulsion to high pressure homogenization to obtain an antibiotic-loaded nanoemulsion.

[0025] When the step 1 is specifically implemented, in order to prepare a nanoemulsion with high efficiency of loading antibiotics, in the preparation method proposed in the present application, an oil phase containing antibiotics is first prepared, and a fat-soluble antibiotic is added to the oil phase to promote the combination of the antibiotic and the oil phase, and enhance the stability and solubility of the antibiotic, so as to achieve high efficiency of loading of the antibiotic in the nanoemulsion. The soybean oil in the oil phase has good solubility and can effectively dissolve fat-soluble substances to achieve high efficiency of loading of antibiotics. At the same time, soybean oil can also be used as a carrier of antibiotics, which helps to release the antibiotics after atomization and inhalation in the body; dipalmitoylphosphatidylcholine, as an emulsifier, has good emulsifying properties and can form a stable interface film between the oil phase and the aqueous phase, thereby preventing the aggregation of oil droplets and promoting the formation of nanoemulsions. At the same time, it can enhance the physical stability of the nanoemulsion and prevent the emulsion from stratification or demulsification during storage and use; cholesterol can improve the biocompatibility of the nanoemulsion, so that it can better interact with the cell membrane in the body and improve the cell adhesion of the drug. The cellular uptake rate will further affect the metabolic pathway and clearance rate of the nanoemulsion in the body, thereby regulating the distribution and residence time of antibiotics in the body and improving the bioavailability and therapeutic effect of the drug; volatile organic solvents can adjust the physical properties of the oil phase, such as viscosity and surface tension. At the same time, during the emulsification process, the volatility of the solvent can affect the stability of the emulsion and the formation of droplets, which can help form smaller and more uniform droplets and improve the stability and performance of the nanoemulsion; further, the interaction between soybean oil and dipalmitoylphosphatidylcholine and cholesterol as emulsifiers can enhance the stability of the nanoemulsion and reduce the leakage and degradation of antibiotics. Soybean oil, dipalmitoylphosphatidylcholine and cholesterol each play an important role in the oil phase of the nanoemulsion, and jointly promote the formation, stability and performance improvement of the nanoemulsion.

[0026] In an optional embodiment of the present application, the volatile organic solvent used in the above step 1 can be at least one of dichloromethane, ethyl acetate, ether and chloroform. Due to the strong solubility of dichloromethane, easy volatility, stable physical and chemical properties and wide application range, the above volatile organic solvent is preferably dichloromethane.

[0027] In an optional embodiment of the present application, the Tween used in step 1 is at least one of Tween 20 and Tween 80. Preferably, the Tween used in step 1 is Tween 80.

[0028] In an optional embodiment of the present application, the antibiotic used in step 1 is preferably a fat-soluble antibiotic, i.e., a hydrophobic antibiotic, which can be selected from at least one of rifampicin, polymyxin and vancomycin, or other hydrophobic antibiotics with equivalent efficacy.

[0029] When the step 2 is specifically implemented, an aqueous phase is prepared by adding an organic acid sodium salt and Tween to a sterile liquid, wherein the organic acid sodium salt can play a role in adjusting the pH value in the aqueous phase, so that the pH value of the nanoemulsion is maintained within an appropriate range, thereby ensuring the stability of the emulsion and the effectiveness of the active ingredient; at the same time, the organic acid sodium salt can be used as a cosolvent to increase the solubility of the active ingredient in the aqueous phase, making it easier to be emulsified and dispersed in the nanoemulsion; secondly, the organic acid sodium salt, as an electrolyte, can affect the double-layer structure and charge distribution in the emulsion system, thereby affecting the stability and particle size distribution of the emulsion. An appropriate amount of electrolyte helps to form a more stable nanoemulsion. Tween has excellent emulsifying properties, can significantly reduce the interfacial tension between the oil phase and the water phase, promote the mixing and emulsification of the oil phase and the water phase, and form a stable nanoemulsion; Tween can be adsorbed on the surface of oil droplets and water droplets to form a protective film to prevent the aggregation and stratification of oil droplets and water droplets, thereby improving the stability of the nanoemulsion; Tween can also increase the solubility of antibiotics or active ingredients in the water phase, making them easier to be emulsified and dispersed in the nanoemulsion, increasing their loading capacity. Therefore, organic acid sodium salt and Tween each play an important role in the water phase of the nanoemulsion, and jointly promote the formation, stability and performance improvement of the nanoemulsion.

[0030] In an optional embodiment of the present application, the sterile liquid used in the above step 2 can be at least one of water, physiological saline, deionized water, phosphate buffer, deoxycholic acid, citrate buffer and Tween aqueous solution, and the Tween aqueous solution contains 0.1 to 5 mg of Tween per 1 ml of sterile liquid.

[0031] In an optional embodiment of the present application, the organic acid sodium salt used in step 2 is at least one of sodium citrate and sodium deoxycholate.

[0032] When the step 3 is implemented specifically, after obtaining the oil phase and the water phase containing the antibiotics respectively, the oil phase and the water phase containing the antibiotics are mixed. In order to fully mix the oil phase and the water phase, considering that ultrasonic treatment has an effective emulsification effect, can promote mixing uniformity, can improve emulsification efficiency, and improve the stability of the emulsion, ultrasonic treatment is used to mix them to obtain a first emulsion. Considering environmental protection and safety, after mixing, the volatile organic solvent is removed by rotary evaporation to obtain a second emulsion. At this time, the particles of the second emulsion mixed by ultrasonic treatment are larger, and the particle size distribution is not uniform. After obtaining the second emulsion, it needs to be subjected to the next step of homogenization.

[0033] In an optional embodiment of the present application, in step 3, the volume ratio of the volatile organic solvent in the oil phase to the sterile liquid in the water phase is 1 to 8:1.

[0034] In a preferred embodiment of the present application, the volume ratio of the volatile organic solvent in the oil phase to the sterile liquid in the water phase is preferably 2-6:1.

[0035] In an optional embodiment of the present application, in step 3, the power of the ultrasonic treatment is 30 watts to 400 watts. Since the power of the ultrasonic treatment is a key factor affecting the effect of nanoemulsion formation, preferably, in step 3, the power of the ultrasonic treatment is 100 watts to 400 watts. In addition, before the ultrasonic treatment, in order to mix the oil phase and the water phase evenly, the oil phase and the water phase are mixed evenly by vortex treatment, and the vortex treatment time is 1 minute to 10 minutes.

[0036] In an optional embodiment of the present application, in step 3, the ultrasonic treatment time is 20 seconds to 20 minutes.

[0037] In an optional embodiment of the present application, in step 3, the temperature of the rotary evaporation to remove the volatile organic solvent in the first emulsion is 37 degrees Celsius to 60 degrees Celsius, and the time of the rotary evaporation to remove the volatile organic solvent in the first emulsion is 5 minutes to 10 minutes.

[0038] When the step 4 is specifically implemented, in order to further improve the stability and homogeneity of the second emulsion, and to obtain a nanoemulsion with a more uniform particle size and a smaller particle size, the second emulsion obtained in step 3 is subjected to high-pressure homogenization to obtain a nanoemulsion loaded with antibiotics. Through further high-pressure homogenization, large particles that may not be fully dispersed by ultrasonic treatment can be broken, so that the emulsion reaches a higher homogeneity, the uniformity of the emulsion particle size can be significantly improved, and an emulsion with a nanoparticle size can be prepared. A smaller particle size distribution helps to reduce the stratification and sedimentation of the emulsion and improve the stability of the nanoemulsion. At the same time, by killing microorganisms with high pressure, the safety of the nanoemulsion is improved and the shelf life is extended, and the active ingredients in the nanoemulsion are retained, and the functionality of the nanoemulsion is improved. Further, compared with solvent-based nanoemulsions, the nanoemulsion loaded with antibiotics prepared in the present application can bring better atomization effects when used for atomization inhalation, and can make the nanoemulsion more retained in the lungs, increase the amount of antibiotics inhaled, and increase the effective concentration of antibiotics in lung lesions.

[0039] In an optional embodiment of the present application, in step 4, the pressure of the high-pressure homogenization treatment is 200 bar to 1000 bar. Since the homogenization pressure is a key factor affecting the nanoemulsion formation effect, preferably, in step 4, the homogenization pressure is 500 bar to 800 bar.

[0040] In an optional embodiment of the present application, in step 4, the time of the high-pressure homogenization treatment is 2 minutes to 20 minutes.

[0041] The preparation method of loaded antibiotics provided in the first aspect of the embodiment of the present application is firstly to mix the antibiotics with the inhalable pharmaceutical excipients soybean oil, DPPC and cholesterol, and dissolve them with a volatile organic solvent to form an oil phase; sodium deoxycholate and Tween are added to a sterile liquid to dissolve to form an aqueous phase; the oil phase and the aqueous phase are then mixed, and subjected to ultrasonic treatment, rotary evaporation treatment and high-pressure homogenization treatment to obtain a nanoemulsion loaded with antibiotics. By specifically adjusting the types of inhalable pharmaceutical excipients, the addition ratio of inhalable pharmaceutical excipients, the ratio of the amount of oil phase and aqueous phase, and the ultrasonic power and homogenization pressure and time, the prepared nanoemulsion can efficiently load antibiotics and has good atomization inhalation performance; higher drug stability, compared with the inhaled antibiotic solution dosage form, the antibiotic nanoemulsion improves the penetration and retention effect of antibiotics in lung tissue, and further improves the ability to resist respiratory infections. The preparation process of this method is simple, and it is prepared by high-pressure homogenization method, which can be mass-produced.

[0042] Based on the same application concept, in a second aspect of the embodiment of the present application, a nanoemulsion loaded with antibiotics is provided. The nanoemulsion loaded with antibiotics is prepared by the preparation method described in any one of the first aspects above.

[0043] The antibiotic-loaded nanoemulsion provided in the embodiment of the present application can enhance the stability and solubility of antibiotics and increase the effective concentration of antibiotics in lung lesions as an inhalable antibiotic drug. Solution-type antibiotics have poor permeability to lung tissue mucus and are easily recognized and cleared by alveolar macrophages with positive charges, which are eliminated quickly. Compared with solution-type antibiotics, nanomaterials can be rationally engineered, such as size control, surface modification, and stimulus-responsive functionalization, thereby producing a unique bactericidal mechanism with bacteria, improving anti-pathogen activity, and facilitating promotion and application.

[0044] Based on the same application concept, in a third aspect of the embodiments of the present application, a use of an antibiotic-loaded nanoemulsion as described in the second aspect above in the preparation of an inhaled antibiotic drug is proposed.

[0045] In order to clearly illustrate the antibiotic-loaded nanoemulsion and its preparation method and application proposed in the present application, a detailed description is given below in conjunction with examples.

[0046] Example 1: Preparation method of a nanoemulsion loaded with antibiotics Example 1A: Preparation of rifampicin-loaded nanoemulsion When the loaded antibiotic is rifampicin, the preparation method of the rifampicin loaded nanoemulsion comprises the following steps: Step 1: 3 mg of rifampicin, 20 mg of soybean oil, 10 mg of DPPC, and 3 mg of cholesterol are mixed uniformly to obtain a mixture; the mixture is dissolved in 1.5 ml of dichloromethane to prepare an oil phase.

[0047] Step 2: Add 3 mg of sodium deoxycholate and 5 mg of Tween 80 to 1 ml of sterile water and stir until the sodium deoxycholate and Tween 80 are dissolved to prepare an aqueous phase.

[0048] Step 3: The volatile organic solvent in the oil phase and the sterile liquid in the water phase are mixed in a volume ratio of 1.5:1, vortex mixed for 3 minutes, and ultrasonicated by a cell disruptor. The power of the ultrasonic treatment is 200 watts, and the ultrasonic treatment time is 8 minutes. After the ultrasonic treatment, a first emulsion is obtained, and the volatile organic solvent in the first emulsion is removed by rotary evaporation using a rotary evaporator to obtain a second emulsion. The rotary evaporation temperature is 45 degrees Celsius, and the rotary evaporation time is 5 minutes.

[0049] Step 4: The second emulsion is subjected to high-pressure homogenization treatment using a high-pressure homogenizer. The pressure of the high-pressure homogenization treatment is 800 bar, the temperature of the high-pressure homogenization is 4 degrees Celsius, and the time of the high-pressure homogenization treatment is 10 minutes to obtain a nanoemulsion loaded with rifampicin.

[0050] Example 1B: Preparation of rifampicin-loaded nanoemulsion When the loaded antibiotic is rifampicin, the preparation method of the rifampicin loaded nanoemulsion comprises the following steps: Step 1: 160 mg of rifampicin, 800 mg of soybean oil, 800 mg of DPPC, and 24 mg of cholesterol are mixed uniformly to obtain a mixture; the mixture is dissolved in chloroform and the volume is fixed to 8 ml to obtain an oil phase, which contains 20 mg / ml of rifampicin, 100 mg / ml of soybean oil, 100 mg / ml of DPPC, and 3 mg / ml of cholesterol.

[0051] Step 2: Add 4 mg of sodium deoxycholate and 5 mg of Tween to 1 ml of sterile water, and stir until the sodium deoxycholate and Tween are dissolved to prepare an aqueous phase containing 5 mg / ml of Tween and 4 mg / ml of sodium deoxycholate.

[0052] Step 3: The mixed material in the oil phase and the sterile liquid in the water phase are mixed in a volume ratio of 8:1, vortex mixed for 10 minutes, and ultrasonicated by a cell disruptor. The ultrasonic treatment power is 400 watts and the ultrasonic treatment time is 20 minutes. After the ultrasonic treatment, a first emulsion is obtained. The volatile organic solvent in the first emulsion is removed by rotary evaporation using a rotary evaporator to obtain a second emulsion. The rotary evaporation temperature is 60 degrees Celsius and the rotary evaporation time is 10 minutes.

[0053] Step 4: The second emulsion is subjected to high-pressure homogenization by a high-pressure homogenizer, the pressure of the high-pressure homogenization is 1000 bar, the temperature of the high-pressure homogenization is 4 degrees Celsius, and the high-pressure homogenization time is 20 minutes to obtain a nanoemulsion loaded with rifampicin.

[0054] Example 1C: Preparation of rifampicin-loaded nanoemulsion When the loaded antibiotic is rifampicin, the preparation method of the rifampicin loaded nanoemulsion comprises the following steps: Step 1: 5 mg of rifampicin, 5 mg of soybean oil, 50 mg of DPPC, and 5 mg of cholesterol are mixed uniformly to obtain a mixture; the mixture is dissolved in 50 ml of chloroform to prepare an oil phase, which contains 0.1 mg / ml of rifampicin, 0.1 mg / ml of soybean oil, 1 mg / ml of DPPC, and 0.1 mg / ml of cholesterol.

[0055] Step 2: Add 5 mg of sodium deoxycholate and 5 mg of Tween into 50 ml of sterile water, and stir until the sodium deoxycholate and Tween are dissolved to prepare an aqueous phase containing 0.1 mg / ml of Tween and 0.1 mg / ml of sodium deoxycholate.

[0056] Step 3: The volatile organic solvent in the oil phase and the sterile liquid in the water phase are mixed in a volume ratio of 1:1, vortex mixed for 1 minute, and ultrasonicated by a cell disruptor. The ultrasonic treatment power is 30 watts, and the ultrasonic treatment time is 20 seconds. After ultrasonic treatment, a first emulsion is obtained, and the volatile organic solvent in the first emulsion is removed by rotary evaporation using a rotary evaporator to obtain a second emulsion. The rotary evaporation temperature is 37 degrees Celsius, and the rotary evaporation time is 5 minutes.

[0057] Step 4: The second emulsion is subjected to high-pressure homogenization by a high-pressure homogenizer, the pressure of the high-pressure homogenization is 200 bar, the temperature of the high-pressure homogenization is 4 degrees Celsius, and the high-pressure homogenization time is 2 minutes to obtain a nanoemulsion loaded with rifampicin.

[0058] Example 1D: Preparation of a Polymyxin-loaded Nanoemulsion When the loaded antibiotic is polymyxin, the preparation method of the nanoemulsion loaded with polymyxin in this embodiment comprises the following steps: Step 1: 3 mg of polymyxin, 20 mg of soybean oil, 10 mg of DPPC, and 3 mg of cholesterol are uniformly mixed to obtain a mixture; the mixture is dissolved in 1.5 ml of dichloromethane to prepare an oil phase.

[0059] Step 2: Add 3 mg of sodium deoxycholate and 5 mg of Tween 80 to 1 ml of sterile water and stir until the sodium deoxycholate and Tween are dissolved to prepare an aqueous phase.

[0060] Step 3: The volatile organic solvent in the oil phase and the sterile liquid in the water phase are mixed in a volume ratio of 1.5:1, vortex mixed for 3 minutes, and ultrasonicated by a cell disruptor. The ultrasonic treatment power is 200 watts, and the ultrasonic treatment time is 8 minutes. After the ultrasonic treatment, a first emulsion is obtained, and the volatile organic solvent in the first emulsion is removed by rotary evaporation to obtain a second emulsion. The rotary evaporation temperature is 45 degrees Celsius, and the rotary evaporation time is 5 minutes.

[0061] Step 4: The second emulsion is subjected to high-pressure homogenization by a high-pressure homogenizer, the pressure of the high-pressure homogenization is 800 bar, the temperature of the high-pressure homogenization is 4 degrees Celsius, and the time of the high-pressure homogenization is 10 minutes to obtain a nanoemulsion loaded with polymyxin.

[0062] Example 1E: Preparation of nanoemulsion loaded with vancomycin When the loaded antibiotic is vancomycin, the preparation method of the nanoemulsion loaded with vancomycin in this embodiment comprises the following steps: Step 1: 3 mg of vancomycin, 20 mg of soybean oil, 10 mg of DPPC, and 3 mg of cholesterol are mixed evenly to obtain a mixture; the mixture is dissolved in 1.5 ml of dichloromethane to prepare an oil phase; Step 2: Add 3 mg of sodium deoxycholate and 5 mg of Tween 80 to 1 ml of sterile water and stir until the sodium deoxycholate and Tween are dissolved to prepare an aqueous phase.

[0063] Step 3: The volatile organic solvent in the oil phase and the sterile liquid in the water phase are mixed in a volume ratio of 1.5:1, vortex mixed for 3 minutes, and ultrasonicated by a cell disruptor. The ultrasonic treatment power is 200 watts, and the ultrasonic treatment time is 8 minutes. After the ultrasonic treatment, a first emulsion is obtained, and the volatile organic solvent in the first emulsion is removed by rotary evaporation to obtain a second emulsion. The rotary evaporation temperature is 45 degrees Celsius, and the rotary evaporation time is 5 minutes.

[0064] Step 4: The second emulsion is subjected to high-pressure homogenization by a high-pressure homogenizer, the pressure of the high-pressure homogenization is 800 bar, the temperature of the high-pressure homogenization is 4 degrees Celsius, and the time of the high-pressure homogenization is 10 minutes to obtain a nanoemulsion loaded with vancomycin.

[0065] In order to illustrate the preparation method proposed in the examples of the present application, by specifically adjusting the type of inhalable pharmaceutical excipients, the addition ratio of the inhalable pharmaceutical excipients, the ratio of the amount of the oil phase and the aqueous phase, and the ultrasonic power and the homogenization pressure and time, the prepared nanoemulsion can efficiently load antibiotics, and has good atomization inhalation performance and high drug stability. The following comparative examples are listed below for comparison with the antibiotic-loaded nanoemulsions prepared by the preparation method proposed in Example 1 (Example 1A-Example 1E).

[0066] Comparative Example 1A: Preparation Method of Nanoemulsion Loaded with Rifampicin Comparative Example 1A Compared with Example 1A, soybean oil was not added, and the remaining operations were the same as those of Example 1A. The specific operations are as follows: Step 1: 3 mg of rifampicin, 10 mg of DPPC, and 3 mg of cholesterol are mixed uniformly to obtain a mixture; the mixture is dissolved in 1.5 ml of dichloromethane to prepare an oil phase.

[0067] Step 2: Add 3 mg of sodium deoxycholate and 5 mg of Tween 80 to 1 ml of sterile water and stir until the sodium deoxycholate and Tween are dissolved to prepare an aqueous phase.

[0068] Step 3: The volatile organic solvent in the oil phase and the sterile liquid in the water phase are mixed in a volume ratio of 1.5:1, vortex mixed for 3 minutes, and ultrasonicated by a cell disruptor. The ultrasonic treatment power is 200 watts, and the ultrasonic treatment time is 8 minutes. After the ultrasonic treatment, a first emulsion is obtained, and the volatile organic solvent in the first emulsion is removed by rotary evaporation to obtain a second emulsion. The rotary evaporation temperature is 45 degrees Celsius, and the rotary evaporation time is 5 minutes.

[0069] Step 4: The second emulsion is subjected to high-pressure homogenization by a high-pressure homogenizer, the pressure of the high-pressure homogenization is 800 bar, the temperature of the high-pressure homogenization is 4 degrees Celsius, and the time of the high-pressure homogenization is 10 minutes to obtain a nanoemulsion loaded with rifampicin.

[0070] Comparative Example 1B: Preparation method of rifampicin-loaded nanoemulsion Comparative Example 1B is compared with Example 1A, except that DPPC is not added, and the remaining operations are the same as those of Example 1A. The specific operations are as follows: Step 1: 3 mg of rifampicin, 20 mg of soybean oil, and 3 mg of cholesterol are mixed evenly to obtain a mixture; the mixture is dissolved in 1.5 ml of dichloromethane to prepare an oil phase.

[0071] Step 2: Add 3 mg of sodium deoxycholate and 5 mg of Tween 80 to 1 ml of sterile water and stir until the sodium deoxycholate and Tween are dissolved to prepare an aqueous phase.

[0072] Step 3: The volatile organic solvent in the oil phase and the sterile liquid in the water phase are mixed in a volume ratio of 1.5:1, vortex mixed for 3 minutes, and ultrasonicated by a cell disruptor. The ultrasonic treatment power is 200 watts, and the ultrasonic treatment time is 8 minutes. After the ultrasonic treatment, a first emulsion is obtained, and the volatile organic solvent in the first emulsion is removed by rotary evaporation to obtain a second emulsion. The rotary evaporation temperature is 45 degrees Celsius, and the rotary evaporation time is 5 minutes.

[0073] Step 4: The second emulsion is subjected to high-pressure homogenization by a high-pressure homogenizer, the pressure of the high-pressure homogenization is 800 bar, the temperature of the high-pressure homogenization is 4 degrees Celsius, and the time of the high-pressure homogenization is 10 minutes to obtain a nanoemulsion loaded with rifampicin.

[0074] Comparative Example 1C: Preparation Method of Nanoemulsion Loaded with Rifampicin Comparative Example 1C is compared with Example 1A, except that cholesterol is not added, and the remaining operations are the same as those of Example 1A. The specific operations are as follows: Step 1: 3 mg of rifampicin, 20 mg of soybean oil, and 10 mg of DPPC are mixed uniformly to obtain a mixture; the mixture is dissolved in 1.5 ml of dichloromethane to prepare an oil phase.

[0075] Step 2: Add 3 mg of sodium deoxycholate and 5 mg of Tween 80 to 1 ml of sterile water and stir until the sodium deoxycholate and Tween are dissolved to prepare an aqueous phase.

[0076] Step 3: Mix the volatile organic solvent in the oil phase and the sterile liquid in the water phase in a volume ratio of 1.5:1, vortex for 3 minutes, and use a cell disruptor to sonicate. The power of the sonication is 200 watts, and the time of the sonication is 8 minutes. After the sonication, a first emulsion is obtained. The volatile organic solvent in the first emulsion is removed by rotary evaporation to obtain a second emulsion. The temperature of the rotary evaporation is 45 degrees Celsius, and the time of the rotary evaporation is 5 minutes.

[0077] Step 4: The second emulsion is subjected to high-pressure homogenization by a high-pressure homogenizer, the pressure of the high-pressure homogenization is 800 bar, the temperature of the high-pressure homogenization is 4 degrees Celsius, and the time of the high-pressure homogenization is 10 minutes to obtain a nanoemulsion loaded with rifampicin.

[0078] Comparative Example 1D: Preparation method of rifampicin-loaded nanoemulsion Comparative Example 1D is compared with Example 1A, the ultrasonic power is changed, and the remaining operations are the same as Example 1A, and the specific operations are as follows: Step 1: 3 mg of rifampicin, 20 mg of soybean oil, 10 mg of DPPC, and 3 mg of cholesterol are mixed uniformly to obtain a mixture; the mixture is dissolved in 1.5 ml of dichloromethane to prepare an oil phase.

[0079] Step 2: Add 3 mg of sodium deoxycholate and 5 mg of Tween 80 to 1 ml of sterile water and stir until the sodium deoxycholate and Tween are dissolved to prepare an aqueous phase.

[0080] Step 3: The volatile organic solvent in the oil phase and the sterile liquid in the water phase are mixed in a volume ratio of 1.5:1, vortex mixed for 3 minutes, and ultrasonicated by a cell disruptor. The ultrasonic treatment power is 450 watts, and the ultrasonic treatment time is 8 minutes. After the ultrasonic treatment, a first emulsion is obtained, and the volatile organic solvent in the first emulsion is removed by rotary evaporation to obtain a second emulsion. The rotary evaporation temperature is 45 degrees Celsius, and the rotary evaporation time is 5 minutes.

[0081] Step 4: The second emulsion is subjected to high-pressure homogenization by a high-pressure homogenizer, the pressure of the high-pressure homogenization is 800 bar, the temperature of the high-pressure homogenization is 4 degrees Celsius, and the homogenization time is 10 minutes to obtain a nanoemulsion loaded with rifampicin.

[0082] Comparative Example 1E: Preparation Method of Nanoemulsion Loaded with Rifampicin Comparative Example 1E is compared with Example 1A, the homogenization pressure is changed, and the remaining operations are the same as Example 1A. The specific operations are as follows: Step 1: 3 mg of rifampicin, 20 mg of soybean oil, 10 mg of DPPC, and 3 mg of cholesterol are mixed uniformly to obtain a mixture; the mixture is dissolved in 1.5 ml of dichloromethane to prepare an oil phase.

[0083] Step 2: Add 3 mg of sodium deoxycholate and 5 mg of Tween 80 to 1 ml of sterile water and stir until the sodium deoxycholate and Tween are dissolved to prepare an aqueous phase.

[0084] Step 3: The volatile organic solvent in the oil phase and the sterile liquid in the water phase are mixed in a volume ratio of 1.5:1, vortex mixed for 3 minutes, and ultrasonicated by a cell disruptor. The ultrasonic treatment power is 200 watts, and the ultrasonic treatment time is 8 minutes. After the ultrasonic treatment, a first emulsion is obtained, and the volatile organic solvent in the first emulsion is removed by rotary evaporation to obtain a second emulsion. The rotary evaporation temperature is 45 degrees Celsius, and the rotary evaporation time is 5 minutes.

[0085] Step 4: The second emulsion is subjected to high-pressure homogenization by a high-pressure homogenizer, the pressure of the high-pressure homogenization is 150 bar, the temperature of the high-pressure average is 4 degrees Celsius, and the time of the high-pressure homogenization is 10 minutes to obtain a nanoemulsion loaded with rifampicin.

[0086] Comparative Example 1F: Preparation method of rifampicin-loaded nanoemulsion Comparative Example 1F Compared with Example 1A, the volume ratio of the volatile organic solvent in the oil phase to the sterile liquid in the water phase was changed to 0.5:1, and the remaining operations were the same as those in Example 1A. The specific operations are as follows: Step 1: 3 mg of rifampicin, 20 mg of soybean oil, 10 mg of DPPC, and 3 mg of cholesterol are mixed uniformly to obtain a mixture; the mixture is dissolved in 1.5 ml of dichloromethane to prepare an oil phase.

[0087] Step 2: Add 3 mg of sodium deoxycholate and 5 mg of Tween 80 to 1 ml of sterile water and stir until the sodium deoxycholate and Tween are dissolved to prepare an aqueous phase.

[0088] Step 3: The volatile organic solvent in the oil phase and the sterile liquid in the water phase are mixed in a volume ratio of 5:1, vortex mixed for 3 minutes, and ultrasonicated by a cell disruptor. The ultrasonic treatment power is 200 watts, and the ultrasonic treatment time is 8 minutes. After ultrasonic treatment, a first emulsion is obtained, and the volatile organic solvent in the first emulsion is removed by rotary evaporation to obtain a second emulsion. The rotary evaporation temperature is 45 degrees Celsius, and the rotary evaporation time is 5 minutes.

[0089] Step 4: The second emulsion is subjected to high-pressure homogenization by a high-pressure homogenizer, the pressure of the high-pressure homogenization is 800 bar, the temperature of the high-pressure homogenization is 4 degrees Celsius, and the time of the high-pressure homogenization is 10 minutes to obtain a nanoemulsion loaded with rifampicin.

[0090] Comparative Example 1G: Preparation Method of Nanoemulsion Loaded with Rifampicin Comparative Example 1G Compared with Example 1A, the volume ratio of the volatile organic solvent in the oil phase to the sterile liquid in the water phase was changed to 10:1, and the remaining operations were the same as those in Example 1A. The specific operations are as follows: Step 1: 3 mg of rifampicin, 20 mg of soybean oil, 10 mg of DPPC, and 3 mg of cholesterol are mixed uniformly to obtain a mixture; the mixture is dissolved in 1.5 ml of dichloromethane to prepare an oil phase.

[0091] Step 2: Add 3 mg of sodium deoxycholate and 5 mg of Tween 80 to 1 ml of sterile water and stir until the sodium deoxycholate and Tween are dissolved to prepare an aqueous phase.

[0092] Step 3: The volatile organic solvent in the oil phase and the sterile liquid in the water phase are mixed in a volume ratio of 10:1, vortex mixed for 3 minutes, and ultrasonicated by a cell disruptor. The power of the ultrasonic treatment is 200 watts, and the ultrasonic treatment time is 8 minutes. After the ultrasonic treatment, a first emulsion is obtained, and the volatile organic solvent in the first emulsion is removed by rotary evaporation to obtain a second emulsion. The rotary evaporation temperature is 45 degrees Celsius, and the rotary evaporation time is 5 minutes.

[0093] Step 4: The second emulsion is subjected to high-pressure homogenization by a high-pressure homogenizer, the pressure of the high-pressure homogenization is 800 bar, the temperature of the high-pressure homogenization is 4 degrees Celsius, and the time of the high-pressure homogenization is 10 minutes to obtain a nanoemulsion loaded with rifampicin.

[0094] Example 2: Performance Characterization of Rifampicin-Loaded Nanoemulsions First, the morphology of the antibiotic-loaded nanoemulsions prepared in Example 1A, Example 1D, and Example 1E was characterized. The characterization results are shown in Figures 1 to 3 The prepared antibiotic-loaded nanoemulsion was subjected to morphological characterization. The characterization results are shown in Figure 1 , Figure 1 is a transmission electron microscopy morphology image of a rifampicin-loaded nanoemulsion provided in an embodiment of the present application, Figure 2 is a transmission electron microscopy morphology of a polymyxin-loaded nanoemulsion provided in an embodiment of the present application, Figure 3 This is a transmission electron microscope morphology of a nanoemulsion loaded with vancomycin provided in an embodiment of the present application. The nanoemulsions loaded with antibiotics rifampicin, polymyxin and vancomycin have uniform morphologies and are all spherical. The experimental method used in this embodiment to characterize the performance of the nanoemulsion loaded with rifampicin is the same as the commonly used technical means in the art, and will not be repeated here.

[0095] Then, the performance of the antibiotic-loaded nanoemulsions prepared in Example 1A to Example 1E and Comparative Example 1A to Comparative Example 1G was characterized, and the encapsulation efficiency, drug loading, particle size, PDI (Polymer Dispersion Index), potential and stability were tested respectively. The test results are shown in Table 1.

[0096] Table 1 Performance characterization results of the antibiotic-loaded nanoemulsions prepared in Example 1A-Example 1E and Comparative Example 1A-Comparative Example 1G

[0097] As can be seen from the results in Table 1, the nanoemulsion loaded with antibiotics prepared by the preparation method of the present application, as an inhalation nanoemulsion, has an efficient loading rate for antibiotics, has good atomization inhalation performance, and has good stability. The ratio of soybean oil, DPPC, and the ratio of sodium deoxycholate and Tween 80 in the nanoemulsion loaded with antibiotics affect the encapsulation rate and drug loading. A larger ultrasonic power can reduce the particle size of the nanoemulsion, but too high an ultrasonic power will affect the physical stability of the nanoemulsion, causing the nanoemulsion to precipitate during storage; sodium deoxycholate can significantly promote the uniformity of the emulsification of the nanoemulsion, and help to form an emulsion with uniform particle size. Without sodium deoxycholate, the nanoemulsion will be difficult to form; the ultrasonic power is too low, the nanoemulsion is not fully emulsified, and it is not easy to form an emulsion.

[0098] Therefore, in the preparation method of the present application, the optimal parameter ratio is that in Example 1A, specifically, the volume ratio of the volatile organic solvent in the oil phase to the sterile liquid in the aqueous phase is 1.5:1, the vortex time is 3 minutes, the power of the ultrasonic treatment is 200 watts, the time of the ultrasonic treatment is 8 minutes, the temperature of the rotary evaporation treatment is 45 degrees Celsius, the time of the rotary evaporation treatment is 5 minutes, the homogenization pressure is 800 bar, and the homogenization time is 10 minutes. By the above preparation method, a nanoemulsion with good stability stored at 4 degrees Celsius for 6 months, a minimum dispersion index of 0.124, and high encapsulation efficiency and drug loading (encapsulation efficiency can reach up to 99%, and drug loading can reach up to 10%) can be obtained.

[0099] Example 3: Characterization of drug delivery efficiency and drug retention of antibiotic-loaded nanoemulsions A nanoemulsion labeled with fluorescent dye DID, i.e., 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindodicarbocyanine perchlorate (1,1′-dioctadecyl-3,3,3′,3′-tetramethylindodicarbocyanine) was selected, and a nanoemulsion loaded with DID-labeled rifampicin (hereinafter referred to as rifampicin nanoemulsion) prepared by the preparation method of Example 1A was used as an inhaled nanoemulsion to observe the distribution of the nanoemulsion into mice.

[0100] The specific operation is as follows: the preparation method of Example 1A is used to prepare a DID-labeled rifampicin nanoemulsion, and intravenous injection of free DID and aerosol inhalation of free DID are used as control groups, respectively. A mouse micro-aerosol inhalation device is used to quantitatively administer an equal amount of DID-labeled nanoemulsion to mice. 48 hours after the mice are administered the aerosol inhalation, the lung tissue of the mice is removed, and the fluorescence content in the lung tissue of each group of mice is observed by a small animal in vivo imaging device, and the drug delivery efficiency and drug retention in the lung tissue are analyzed.

[0101] The above experimental results refer to Figure 4 , Figure 4 This is a quantitative detection diagram of lung tissue of a rifampicin nanoemulsion provided in an example of the present application 48 hours after inhalation by mice. Figure 4 DID represents the distribution of the drug (i.e., rifampicin nanoemulsion) in mice. In the mouse model, after rifampicin nanoemulsion was inhaled by mice, the fluorescence signal of the lung tissue of the nanoemulsion group (i.e., DID-labeled nanoemulsion) was significantly higher than that of the intravenous injection group and the nebulized inhalation group of free DID. The exposure of DID-labeled rifampicin nanoemulsion after nebulized inhalation at 48 hours was significantly higher than that of inhalation administration (nebulized inhalation DID) and intravenous administration (intravenous injection DID), indicating that nebulized inhalation of rifampicin nanoemulsion can significantly improve the efficiency of drug delivery in the lungs, and has the advantages of increasing the drug retention and exposure time in lung tissue.

[0102] The above is a detailed introduction to an antibiotic-loaded nanoemulsion, a preparation method and an application thereof provided by the present application. Specific examples are used herein to illustrate the principles and implementation methods of the present application. The description of the above embodiments is only used to help understand the method and core idea of ​​the present application. At the same time, for those skilled in the art, according to the idea of ​​the present application, there will be changes in the specific implementation methods and application scopes. In summary, the content of this specification should not be understood as limiting the present application.

Claims

1. A method for preparing an antibiotic-loaded nanoemulsion, characterized in that: The method comprises the following steps: Step 1: Evenly mix antibiotics, soybean oil, dipalmitoylphosphatidylcholine and cholesterol to obtain a mixture, and dissolve the mixture in a volatile organic solvent to prepare an oil phase, wherein 0.1 to 20 mg of antibiotics, 0.1 to 100 mg of soybean oil, 1 to 100 mg of dipalmitoylphosphatidylcholine and 0.1 to 3 mg of cholesterol are added to each 1 ml of the volatile organic solvent in the oil phase; Step 2: adding an organic acid sodium salt and Tween to a sterile liquid, stirring until the organic acid sodium salt and the Tween are dissolved, to prepare an aqueous phase, wherein 0.1 to 4 mg of the organic acid sodium salt and 0.1 to 5 mg of Tween are added to each 1 ml of the sterile liquid in the aqueous phase; Step 3: After the oil phase and the water phase are uniformly mixed, ultrasonic treatment is performed to obtain a first emulsion, and the volatile organic solvent in the first emulsion is removed by rotary evaporation to obtain a second emulsion, wherein the mixing volume ratio of the volatile organic solvent in the oil phase to the sterile liquid in the water phase is 1 to 8:1; Step 4: subjecting the second emulsion to high pressure homogenization to obtain an antibiotic-loaded nanoemulsion.

2. The method for preparing the antibiotic-loaded nanoemulsion according to claim 1, characterized in that: In step 1, the volatile organic solvent is at least one of dichloromethane, ethyl acetate, ether and chloroform.

3. The method for preparing the antibiotic-loaded nanoemulsion according to claim 1, characterized in that: In step 1, the antibiotic is at least one of rifampicin, polymyxin and vancomycin.

4. The method for preparing the antibiotic-loaded nanoemulsion according to claim 1, characterized in that: In step 2, the sterile liquid is at least one of water, physiological saline, deionized water, phosphate buffer, deoxycholic acid, citrate buffer and Tween aqueous solution.

5. The method for preparing the antibiotic-loaded nanoemulsion according to claim 1, characterized in that: In step 2, the organic acid sodium salt is at least one of sodium citrate and sodium deoxycholate.

6. The method for preparing the antibiotic-loaded nanoemulsion according to claim 1, characterized in that: In step 3, the power of the ultrasonic treatment is 30 watts to 400 watts, and the time of the ultrasonic treatment is 20 seconds to 20 minutes.

7. The method for preparing the antibiotic-loaded nanoemulsion according to claim 1, characterized in that: In the step 3, the temperature of the rotary evaporation to remove the volatile organic solvent in the first emulsion is 37 degrees Celsius to 60 degrees Celsius, and the time of the rotary evaporation to remove the volatile organic solvent in the first emulsion is 5 minutes to 10 minutes.

8. The method for preparing the antibiotic-loaded nanoemulsion according to claim 1, characterized in that: In step 4, the pressure of the high-pressure homogenization treatment is 200 bar to 1000 bar, and the time of the high-pressure homogenization treatment is 2 minutes to 20 minutes.

9. A nanoemulsion loaded with antibiotics, characterized in that: The antibiotic-loaded nanoemulsion is prepared by the preparation method according to any one of claims 1 to 8.

10. Use of the antibiotic-loaded nanoemulsion according to claim 9 in the preparation of inhaled antibiotic drugs.

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