A kind of nano-drug for inhalation therapy of pulmonary pseudomonas aeruginosa infection and its preparation method

Abstract

This invention discloses a nanomedicine for inhalation therapy of Pseudomonas aeruginosa infection in the lungs and its preparation method, belonging to the field of biomaterials technology. It includes tobramycin, nanozymes, and a liposome carrier; the liposome carrier is composed of 1,2-dioleoyl-sn-glycerol-3-phosphoethanolamine, cholesterol hemisuccinate, and distearate phosphatidylethanolamine-polyethylene glycol 2000. The nanomedicine of this invention can penetrate the lung mucus layer and disrupt the Pseudomonas aeruginosa biofilm, directly delivering the drug to the infected area of ​​the lungs using nebulized inhalation technology, solving the problems faced in clinical oral or intravenous antibiotic treatment of Pseudomonas aeruginosa infection in the lungs. Furthermore, it exhibits high safety and stability, and the preparation process is simple and reasonable, creating conditions for industrial commercialization.

Description

Technical Field

[0001] This invention relates to the field of biomaterials technology, and in particular to a nanomedicine for inhalation therapy of Pseudomonas aeruginosa infection of the lungs and its preparation method. Background Technology

[0002] Lung infections are common respiratory diseases characterized by high morbidity and mortality, posing a significant threat to public health and increasing the social burden. *Pseudomonas aeruginosa* is a common pathogen causing lung infections; it is easily colonized, highly drug-resistant, and forms biofilms after infection, making it difficult to clear. Furthermore, *Pseudomonas aeruginosa* infection stimulates lower respiratory tract mucosal cells to secrete more mucus, leading to a thickening of the mucus layer.

[0003] The current clinical treatment for Pseudomonas aeruginosa infection in the lungs mainly includes oral or intravenous antibiotics, which require high doses to achieve effective drug concentrations in lung tissue, leading to serious side effects.

[0004] Compared to traditional treatments, nebulized drug delivery has received more attention in recent years because it can deliver antibiotics directly to the infected area, showing better therapeutic effects with lower doses. Several inhaled formulations of antibiotics have already been used clinically. Tobramycin inhalation solution is the world's first inhaled formulation for treating bronchiectasis with Pseudomonas aeruginosa colonization, and also the first inhaled antibiotic approved for marketing in my country. It is the primary drug for inhaled treatment of lung infections and for clearing Pseudomonas aeruginosa colonization in the lungs. However, the biofilm formed by Pseudomonas aeruginosa is its main resistance mechanism, making it difficult for antibiotics to clear. The thickening of the pulmonary mucus layer caused by infection also poses challenges to inhaled antibiotic therapy. Furthermore, the release of the drug cannot be controlled during inhalation, all of which affect the efficacy of inhaled therapy.

[0005] Liposomes, as classic carriers of nanomaterials, are widely used in nanomedicine delivery systems. Inhaled liposomal drugs have emerged, but research in the treatment of pulmonary Pseudomonas aeruginosa infections is extremely limited. Furthermore, liposomes can be modified with PEG. PEG is a highly hydrophilic polymer with extremely low toxicity, high biocompatibility, and is non-immunogenic, exhibiting low protein adsorption capacity. Since the main component of the lower respiratory tract mucus layer is mucin, PEG modification can reduce the interaction between liposomes and the mucus layer, increasing their fluidity in the mucus and thus facilitating diffusion. Moreover, lipid components unstable in acidic environments can be used during preparation, endowing liposomes with the ability to release drugs in the acidic microenvironment of lung infection. Loading fluorescent materials into liposomes allows for real-time tracking of the distribution and release of liposomal drugs in the infected lung area using fluorescence imaging.

[0006] Nanozymes, a popular material in nanotechnology research in recent years, possess catalytic activity similar to natural enzymes, relatively small size, and high environmental stability. They can penetrate biofilms, effectively degrading them, and have begun to be used for antibacterial purposes. Metallic materials, non-metallic materials, or their compounds and hybrids can all be used to prepare nanozymes. Nanozymes can exhibit different enzymatic activities under different conditions, thereby disrupting biofilms through various mechanisms. For example, nanozymes with peroxidase-like activity can effectively catalyze the conversion of hydrogen peroxide (H₂O₂) into highly reactive oxygen species, such as hydroxyl radicals (·OH), inactivating resident microorganisms and thus eradicating the entire biofilm. Summary of the Invention

[0007] The purpose of this invention is to provide a nanomedicine for inhalation therapy of Pseudomonas aeruginosa infection in the lungs and its preparation method, so as to solve the problems existing in the prior art.

[0008] To achieve the above objectives, the present invention provides the following solution: One of the technical solutions of the present invention is a nanomedicine for inhalation therapy of Pseudomonas aeruginosa infection in the lungs, comprising tobramycin, nanozyme and liposome carrier; The liposome carrier is composed of 1,2-dioleoyl-sn-glycerol-3-phosphoethanolamine, cholesterol hemisuccinate, and distearate phosphatidylethanolamine-polyethylene glycol 2000.

[0009] The second technical solution of the present invention is a method for preparing the aforementioned nanomedicine, comprising the following steps: (1) 1,2-dioleoyl-sn-glycerol-3-phosphoethanolamine, cholesterol hemisuccinate and distearate phosphatidylethanolamine-polyethylene glycol 2000 were mixed in proportion as liposome carriers, dissolved in organic solvents, and the organic solvents were removed by rotary evaporation under reduced pressure to form a film. (2) Add a mixed solution of tobramycin and Mn3O4 to the film for hydration, and then treat it with ultrasound to obtain the nanomedicine.

[0010] The third technical solution of the present invention is the application of the nanomedicine in the preparation of a drug for inhalation therapy of Pseudomonas aeruginosa infection of the lungs.

[0011] The fourth technical solution of the present invention is an inhaled drug for treating Pseudomonas aeruginosa infection of the lungs, comprising the aforementioned nanomedicine.

[0012] Based on the above technical solution, the present invention has the following technical effects: (1) This product is a liposome. The phospholipid bilayer of the liposome is similar to that of mammalian cell membranes, which can reduce the body's immune response and thus reduce drug toxicity. By using lipids that are unstable under acidic conditions, the liposomes can disintegrate in the acidic microenvironment of lung infection, thereby achieving controlled drug release, which is beneficial to improving the therapeutic effect and reducing side effects.

[0013] (2) Polyethylene glycol (PEG) has a low protein adsorption capacity, while the main component of the mucus layer of the lower respiratory tract is mucin. Therefore, PEG modification can reduce the interaction between liposomes and the mucus layer, increase their fluidity in the mucus, and make it easier for liposomes to diffuse in the mucus.

[0014] (3) Mn3O4 can act as a nanozyme, with catalytic activity similar to natural enzymes, relatively small size and high environmental stability, and can enter the interior of biofilm, thereby effectively degrading biofilm.

[0015] (4) This product can solve the problems faced in the clinical treatment of pulmonary Pseudomonas aeruginosa infection by nebulized tobramycin from three aspects: penetrating the pulmonary mucus barrier, destroying the biofilm of Pseudomonas aeruginosa, and controlling drug release, thereby achieving highly effective treatment.

[0016] (5) The present invention synthesizes a novel nanoparticle with good stability and safety. The preparation process is simple and reasonable, and it can be industrialized. It has broad prospects in the medical field.

[0017] The nanomedicine of this invention can penetrate the lung mucus layer and disrupt the biofilm of *Pseudomonas aeruginosa*. Utilizing nebulized inhalation technology, the drug is directly delivered to the infected area of ​​the lungs, solving the problems encountered in clinical oral or intravenous antibiotic treatment of *Pseudomonas aeruginosa* infections of the lungs. Furthermore, it exhibits high safety and stability, and its preparation process is simple and rational, paving the way for industrial commercialization. Attached Figure Description

[0018] Figure 1 This is a transmission electron microscope (TEM) image of Tob-Mn3O4-Lip nanoparticles.

[0019] Figure 2 The hydrated particle size of Tob-Mn3O4-Lip nanoparticles under dynamic light scattering.

[0020] Figure 3 The potential of Tob-Mn3O4-Lip nanoparticles under dynamic light scattering.

[0021] Figure 4 EDS elemental analysis of Tob-Mn3O4-Lip nanoparticles.

[0022] Figure 5The study evaluates the bactericidal effect of Tob-Mn3O4-Lip nanoparticles on the standard strain PAO1 of Pseudomonas aeruginosa. In this study, a represents the minimum inhibitory concentration (MIC) of Tob-Mn3O4-Lip nanoparticles against PAO1, b represents a comparison of the antibacterial effects of Tob-Mn3O4-Lip nanoparticles with other drugs such as Tob, and c represents the survival or death of confocal bacteria after drug treatment.

[0023] Figure 6 The interaction between Tob-Mn3O4-Lip nanoparticles and Pseudomonas aeruginosa biofilm.

[0024] Figure 7 This refers to the ability of Tob-Mn3O4-Lip nanoparticles to penetrate the lung mucus layer.

[0025] Figure 8 The pH responsiveness of Tob-Mn3O4-Lip nanoparticles.

[0026] Figure 9 Tob-Mn3O4-Lip nanoparticles exhibit good biocompatibility.

[0027] Figure 10 This study investigated the safety and therapeutic efficacy of Tob-Mn3O4-Lip nanoparticles. Specifically, a represents the change in body temperature of mice before and after inhalation treatment; b represents the change in body weight of mice before and after inhalation treatment; c represents the bacterial load in lung tissue homogenate and bronchoalveolar lavage fluid of mice after different drug treatments; and d represents the liver and kidney function tests of mice after different drug treatments. Detailed Implementation

[0028] Unless otherwise specified, the technical solutions described in this invention are all conventional solutions in the field, and the reagents or raw materials used are all purchased from commercial channels or are publicly available unless otherwise specified.

[0029] This invention provides a nanomedicine for inhalation therapy of Pseudomonas aeruginosa infection in the lungs, comprising tobramycin, nanozyme, and liposome carrier; The liposome carrier is composed of 1,2-dioleoyl-sn-glycerol-3-phosphoethanolamine, cholesterol hemisuccinate, and distearate phosphatidylethanolamine-polyethylene glycol 2000.

[0030] In some specific implementations, the nanozyme is Mn3O4 nanoparticles.

[0031] In some specific embodiments, the nanomedicine comprises, by weight percentage: 30-35% tobramycin, 3-5% Mn3O4 nanoparticles, 40-50% 1,2-dioleoyl-sn-glycerol-3-phosphoethanolamine, 10-15% cholesterol hemisuccinate, and 5-10% distearylphosphatidylethanolamine-polyethylene glycol 2000.

[0032] In some specific embodiments, the nanomedicine comprises, by weight percentage: 32.25% tobramycin, 3.22% Mn3O4 nanoparticles, 45.16% 1,2-dioleoyl-sn-glycerol-3-phosphoethanolamine, 12.9% cholesterol hemisuccinate, and 6.47% distearylphosphatidylethanolamine-polyethylene glycol 2000.

[0033] This invention also provides a method for preparing the nanomedicine, comprising the following steps: (1) 1,2-dioleoyl-sn-glycerol-3-phosphoethanolamine, cholesterol hemisuccinate and distearate phosphatidylethanolamine-polyethylene glycol 2000 were mixed in proportion as liposome carriers, dissolved in organic solvents, and the organic solvents were removed by rotary evaporation under reduced pressure to form a film. (2) Add a mixed solution of tobramycin and Mn3O4 to the film for hydration, and then treat it with ultrasound to obtain the nanomedicine.

[0034] In some specific implementations, the conditions for ultrasonic treatment are: low-temperature ultrasonic fragmentation, ultrasonic treatment for 1 second followed by a 1-second pause, total ultrasonic power of 130W, ultrasonic power of 60%, and total ultrasonic time of 6 minutes.

[0035] This invention also provides the application of the nanomedicine in the preparation of a medicament for inhalation therapy of Pseudomonas aeruginosa infection of the lungs.

[0036] This invention also provides an inhaled treatment for Pseudomonas aeruginosa infection of the lungs, including the aforementioned nanomedicine.

[0037] The nanomedicine of this invention can penetrate the lung mucus layer and disrupt the biofilm of *Pseudomonas aeruginosa*. Utilizing nebulized inhalation technology, the drug is directly delivered to the infected area of ​​the lungs, solving the problems encountered in clinical oral or intravenous antibiotic treatment of *Pseudomonas aeruginosa* infections of the lungs. Furthermore, it exhibits high safety and stability, and its preparation process is simple and rational, paving the way for industrial commercialization.

[0038] By co-loading tobramycin and nanozymes into liposomes, a nanomedicine for inhalation therapy of Pseudomonas aeruginosa infection in the lungs can be prepared. In the acidic environment of the lungs, the liposomes, modified with PEG, penetrate the mucus barrier, undergo pH-responsive disintegration, and release tobramycin and nanozymes. Tobramycin acts as an antibiotic to inhibit bacteria, while the nanozymes disrupt the biofilm of Pseudomonas aeruginosa, thus enhancing the therapeutic effect of Pseudomonas aeruginosa infection in the lungs from multiple aspects and also facilitating nebulized inhalation. Developing an antibiotic delivery system with the ability to disrupt the Pseudomonas aeruginosa biofilm, penetrate the mucus layer, and release drugs responsively addresses key issues in current treatment. Furthermore, fluorescence imaging can track the distribution and release of the drug in the infected lung area in real time, which is beneficial for the diagnosis and treatment of Pseudomonas aeruginosa infection in the lungs.

[0039] In this invention, nanoparticles are delivered to the lungs via nebulized intratracheal inoculation. This delivery method is a local drug delivery approach, directly delivering the particles to the lungs without requiring specific targeting molecules. Instead, it relies on physical delivery. Therefore, the nanoparticles prepared in this invention are delivered directly to the site of lung infection via nebulized intratracheal inoculation. This local drug delivery method utilizes the physiological characteristics of the respiratory tract to enable the nanoparticles to deposit efficiently in the lungs.

[0040] Example 1 A nanomedicine for treating pulmonary Pseudomonas aeruginosa infection comprises the following components by weight percentage of the total raw materials: Tobramycin 32.25%, Mn3O4 nanoparticles 3.22%, DOPE (1,2-dioleoyl-sn-glycerol-3-phosphoethanolamine) 45.16%, CHEMS (cholesterol hemisuccinate) 12.9%, DSPE-mPEG 2000 (Distearylphosphatidylethanolamine-polyethylene glycol 2000) 6.47%.

[0041] Preparation method: (1) Manganese acetate (0.17 g), oleic acid (640 µL), and oleylamine (3.28 mL) were mixed and dissolved in xylene (15 mL). The mixture was stirred at 90 °C for 10 minutes, and then deionized water (1 mL) was added. The reaction was allowed to proceed for 2.5 hours. After the reaction, anhydrous ethanol (40 mL) was added, and the precipitate was collected by centrifugation and dispersed in cyclohexane (1 mL) to obtain Mn3O4 nanoparticles.

[0042] (2) Add 300 µL of Tween 20 dropwise to 20 mL of deionized water and stir at room temperature for 30 min to mix evenly. Take 15.6 mg of Mn3O4 nanoparticles synthesized by high-temperature pyrolysis, dissolve them in 1 mL of cyclohexane, and take 500 µL of the solution in deionized water containing Tween 20. Sonicate in a water bath for 1 h until the solution is clear. Add 2 mL of dopamine hydrochloride solution (1 mg / mL) and stir overnight at room temperature. Centrifuge at 14000 rpm and 4℃ for 2 h, collect the precipitate, and disperse it in 1.5 mL of deionized water to obtain polydopamine (PDA) coated Mn3O4, i.e., Mn3O4 solution, so that the water-insoluble Mn3O4 can be dissolved in water with the help of the PDA shell.

[0043] (3) Weigh out DOPE (1,2-dioleoyl-sn-glycerol-3-phosphoethanolamine), CHEMS (cholesterol hemisuccinate), and DSPE-mPEG. 2000 A mixture of 10.86 mg of distearate phosphatidylethanolamine and polyethylene glycol 2000 (molar ratio 3:2:0.15) was used as a liposome carrier, dissolved in chloroform, and dissolved by sonication in a 50 mL round-bottom flask. The organic solvent was removed by rotary evaporation under reduced pressure at 40 °C to form a uniform film. 5 mL of tobramycin solution (1 mg / mL) and 1 mL of Mn3O4 solution (0.5 mg / mL) were sonicated and added to the round-bottom flask. The mixture was magnetically stirred in a 37 °C water bath to ensure complete hydration (approximately 1 h), followed by low-temperature sonication for 6 min (1 s sonication, 1 s pause, total sonication power 130 W, sonication power 60%) to obtain the Tob-Mn3O4-Lip solution.

[0044] Example 2 A nanomedicine for treating pulmonary Pseudomonas aeruginosa infection comprises the following components by weight percentage of the total raw materials: Tobramycin 32.25%, Mn3O4 nanoparticles 3.22%, DOPE (1,2-dioleoyl-sn-glycerol-3-phosphoethanolamine) 45.16%, CHEMS (cholesterol hemisuccinate) 12.9%, DSPE-mPEG 2000 (Distearylphosphatidylethanolamine-polyethylene glycol 2000) 6.47%.

[0045] Preparation method: (1) Manganese acetate (0.17 g), oleic acid (640 µL), and oleylamine (3.28 mL) were mixed and dissolved in xylene (15 mL). The mixture was stirred at 90 °C for 10 minutes, and then deionized water (1 mL) was added. The reaction was allowed to proceed for 3 hours. After the reaction, anhydrous ethanol (40 mL) was added, and the precipitate was collected by centrifugation and freeze-dried to obtain Mn3O4 nanoparticles.

[0046] (2) Add 300 µL of Tween 20 dropwise to 20 mL of deionized water and stir at room temperature for 30 min to mix evenly. Take 15.6 mg of Mn3O4 nanoparticles synthesized by high-temperature pyrolysis, dissolve them in 1 mL of cyclohexane, and take 500 µL of the solution in deionized water containing Tween 20. Sonicate in a water bath for 1 h until the solution is clear. Add 2 mL of dopamine hydrochloride solution (1 mg / mL) and stir overnight at room temperature. Centrifuge at 14000 rpm and 4℃ for 2 h, collect the precipitate, and disperse it in 1.5 mL of deionized water to obtain polydopamine (PDA) coated Mn3O4, i.e., Mn3O4 solution, so that the water-insoluble Mn3O4 can be dissolved in water with the help of the PDA shell.

[0047] (3) Weigh DOPE, CHEMS, DSPE-mPEG 2000 A mixture of 10.86 mg (molar ratio of the three components: 3:2:0.15) was used as a liposome carrier, dissolved in chloroform organic solvent, and placed in a 50 mL round-bottom flask for ultrasonic dissolution. The organic solvent was removed by rotary evaporation under reduced pressure at 40 °C to form a uniform film. 10 mL of tobramycin solution (1 mg / mL) and 1 mL of Mn3O4 solution (1 mg / mL) were ultrasonically mixed and added to the round-bottom flask. The mixture was magnetically stirred in a 37 °C water bath to ensure complete hydration (about 1 h). The mixture was then subjected to low-temperature ultrasonic disruption for 6 min (1 s of ultrasonication followed by 1 s of pause, total ultrasonic power of 130 W, ultrasonic power of 60%) to obtain the Tob-Mn3O4-Lip solution.

[0048] Example 3 A nanomedicine for treating pulmonary Pseudomonas aeruginosa infection comprises the following components by weight percentage of the total raw materials: Tobramycin 32.25%, Mn3O4 nanoparticles 3.22%, DOPE (1,2-dioleoyl-sn-glycerol-3-phosphoethanolamine) 45.16%, CHEMS (cholesterol hemisuccinate) 12.9%, DSPE-mPEG 2000 (Distearylphosphatidylethanolamine-polyethylene glycol 2000) 6.47%.

[0049] Preparation method: (1) Weigh 0.613 g of manganese acetate tetrahydrate (Mn(CH3COO)2·4H2O), add 30 mL of anhydrous ethanol to dissolve it completely, place it in a polytetrafluoroethylene-lined reactor and seal it. Place the sealed reactor on a constant temperature magnetic stirrer, and while maintaining magnetic stirring, raise the temperature to 100 ℃ (5 ℃ / min), maintain this temperature and stir for 24 h. After that, let it cool naturally to room temperature, transfer the solution in the reactor to a centrifuge tube, centrifuge at 4000 rpm for 10 min to separate the product, discard the supernatant, wash the obtained precipitate three times with anhydrous methanol, and place it in an electric vacuum drying oven at 60 ℃ for 12 h. The final brown powder obtained is Mn3O4 nanoparticles.

[0050] (2) Add 300 µL of Tween 20 dropwise to 20 mL of deionized water and stir at room temperature for 30 min to mix evenly. Take 15.6 mg of Mn3O4 nanoparticles synthesized by high-temperature pyrolysis, dissolve them in 1 mL of cyclohexane, and take 500 µL of the solution in deionized water containing Tween 20. Sonicate in a water bath for 1 h until the solution is clear. Add 2 mL of dopamine hydrochloride solution (1 mg / mL) and stir overnight at room temperature. Centrifuge at 14000 rpm and 4℃ for 2 h, collect the precipitate, and disperse it in 1.5 mL of deionized water to obtain polydopamine (PDA) coated Mn3O4, i.e., Mn3O4 solution, so that the water-insoluble Mn3O4 can be dissolved in water with the help of the PDA shell.

[0051] (3) Weigh DOPE, CHEMS, DSPE-mPEG 2000 A mixture of 10.86 mg (molar ratio of the three components: 3:2:0.15) was used as a liposome carrier, dissolved in chloroform organic solvent, and placed in a 50 mL round-bottom flask for ultrasonic dissolution. The organic solvent was removed by rotary evaporation under reduced pressure at 40 °C to form a uniform film. 10 mL of tobramycin solution (1 mg / mL) and 1 mL of Mn3O4 solution (1 mg / mL) were ultrasonically mixed and added to the round-bottom flask. The mixture was magnetically stirred in a 37 °C water bath to ensure complete hydration (about 1 h). The mixture was then subjected to low-temperature ultrasonic disruption for 6 min (1 s of ultrasonication followed by 1 s of pause, total ultrasonic power of 130 W, ultrasonic power of 60%) to obtain the Tob-Mn3O4-Lip solution.

[0052] Example of effect 1. To test the bactericidal effect of the Tob-Mn3O4-Lip nanoparticles synthesized in Example 1, they were applied to the Pseudomonas aeruginosa standard strain PAO1 to verify their bactericidal effect. The results are as follows: Figure 5 As shown, Figure 5 In the figure, 'a' represents the minimum inhibitory concentration (MIC) of Tob-Mn3O4-Lip nanoparticles against PAO1, which is 16 µg / mL. Figure 5 In Figure b, the antibacterial effect of Tob-Mn3O4-Lip nanoparticles is compared with that of other drugs such as Tob. Figure 5 In the middle (c), the results show the viability and mortality of confocal bacteria after drug application. The results indicate that Tob-Mn3O4-Lip nanoparticles have a better antibacterial effect than Tob alone, demonstrating the synergistic antibacterial effect of Tob and Mn3O4.

[0053] 2. To verify the interaction between the Tob-Mn3O4-Lip nanoparticles synthesized in Example 1 and the Pseudomonas aeruginosa biofilm, the three-dimensional structure of the biofilm after interaction with the PAO1 biofilm was captured using a confocal microscope, as shown in the figure. Figure 6 As shown, Tob-Mn3O4-Lip nanoparticles can scavenge biofilms.

[0054] 3. To verify the ability of the Tob-Mn3O4-Lip nanoparticles synthesized in Example 1 to penetrate the lung mucus layer, they were reacted with the mucus layer of an artificial lung, as follows: Figure 7 As shown, this demonstrates that Tob-Mn3O4-Lip nanoparticles have the ability to penetrate the mucus layer.

[0055] 4. To verify the pH responsiveness of the Tob-Mn3O4-Lip nanoparticles synthesized in Example 1, the tobramycin release rate at different pH values ​​was measured, such as... Figure 8 As shown, it is demonstrated that the release is fastest in an acidic environment (pH=5.5), indicating that the nanoparticles have pH responsiveness.

[0056] 5. The Tob-Mn3O4-Lip nanoparticles synthesized in Example 1 exhibit good biocompatibility, such as... Figure 9 As shown, the CCK-8 cytotoxicity assay demonstrated that it is essentially non-toxic to normal cells.

[0057] 6. Verify the safety and therapeutic efficacy of the Tob-Mn3O4-Lip nanoparticles synthesized in Example 1 at the animal level, such as... Figure 10 As shown, Figure 10 In the middle, 'a' represents the change in body temperature of mice before and after treatment. Figure 10 b represents weight change. Figure 10 In the middle (c), we see the bacterial agar plate distribution of lung tissue homogenate and bronchoalveolar lavage fluid. Figure 10 d represents liver and kidney function tests after treatment. This demonstrates that the Tob-Mn3O4-Lip nanoparticles synthesized in Example 1 have good therapeutic effects and are safe and non-toxic in animal studies.

[0058] Obviously, the above embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the implementation of the present invention. For those skilled in the art, other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively describe all embodiments here. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention should be included within the scope of protection of the claims of the present invention.

Claims

1. A nano-drug for inhalation therapy of pulmonary Pseudomonas aeruginosa infection, characterized in that, Including tobramycin, nanozymes, and liposome carriers; The liposome carrier is composed of 1,2-dioleoyl-sn-glycerol-3-phosphoethanolamine, cholesterol hemisuccinate, and distearate phosphatidylethanolamine-polyethylene glycol 2000.

2. The nanomedicine according to claim 1, wherein, The nanozyme is Mn3O4 nanoparticles.

3. The nanomedicine according to claim 2, wherein, The nanomedicine comprises, by weight percentage: 30-35% tobramycin, 3-5% Mn3O4 nanoparticles, 40-50% 1,2-dioleoyl-sn-glycerol-3-phosphoethanolamine, 10-15% cholesterol hemisuccinate, and 5-10% distearylphosphatidylethanolamine-polyethylene glycol 2000.

4. The nanomedicine according to claim 3, wherein, The nanomedicine comprises, by weight percentage: tobramycin 32.25%, Mn3O4 nanoparticles 3.22%, 1,2-dioleoyl-sn-glycerol-3-phosphoethanolamine 45.16%, cholesterol hemisuccinate 12.9%, and distearylphosphatidylethanolamine-polyethylene glycol 2000 6.47%.

5. A method for preparing the nanodrug according to any one of claims 1 to 4, characterized in that, Includes the following steps: (1) 1,2-dioleoyl-sn-glycerol-3-phosphoethanolamine, cholesterol hemisuccinate and distearate phosphatidylethanolamine-polyethylene glycol 2000 were mixed in proportion as liposome carriers, dissolved in organic solvents, and the organic solvents were removed by rotary evaporation under reduced pressure to form a film. (2) Add a mixed solution of tobramycin and Mn3O4 to the film for hydration, and then treat it with ultrasound to obtain the nanomedicine.

6. The production method according to claim 5, wherein The conditions for ultrasonic treatment are: low-temperature ultrasonic fragmentation, ultrasonic treatment for 1 second followed by a 1-second pause, total ultrasonic power of 130W, ultrasonic power of 60%, and total ultrasonic time of 6 minutes.

7. The use of the nanomedicine according to any one of claims 1-4 in the preparation of a medicament for inhalation therapy of Pseudomonas aeruginosa infection of the lungs.

8. A medicament for inhalation therapy of pulmonary Pseudomonas aeruginosa infection, characterized in that, Including the nanomedicines according to any one of claims 1-4.

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