Glutathione dry powder inhalant as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of antioxidant drugs, and particularly relates to a glutathione dry powder inhalant as well as a preparation method and application thereof. The glutathione dry powder inhalant is prepared from the following active ingredients in percentage by mass: 50%-60% of reduced glutathione, 10%-20% of a carrier and 20%-30% of a dispersing agent, totaling 100%, the carrier is mannitol, lactose or trehalose, the dispersing agent is leucine, and no other pharmaceutic adjuvants exist. The glutathione dry powder inhalant is obtained by uniformly mixing the raw materials of the glutathione dry powder inhalant and then carrying out spray drying.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of antioxidant drugs, and particularly relates to a glutathione dry powder inhalant as well as a preparation method and application thereof. BACKGROUND

[0002] Asthma is a chronic airway inflammatory disease characterized by airway hyperresponsiveness and variable airflow limitation, mainly manifested as repeated attacks of wheezing, shortness of breath, chest tightness and cough. In clinical treatment drugs, reduced glutathione as an endogenous antioxidant tripeptide has unique potential in asthma treatment due to its antioxidant, protection of airway epithelial cells and enhancement of immune function.

[0003] The glutathione dosage forms currently used in clinical applications are mainly oral tablets and injections, such as reduced glutathione tablets and reduced glutathione for injection. Among them, the oral preparation has low bioavailability, and the injection is inconvenient to administer, resulting in poor long-term medication compliance of patients, and both of them need a large dose to maintain the effective treatment concentration of lung lesions, which can easily cause systemic adverse reactions. However, in some cases of acute asthma attack, the traditional oral preparations and injection forms are difficult to meet the needs of rapid local drug delivery. Dry powder inhalants can quickly take effect, are convenient to carry, and are suitable for long-term medication of asthma.

[0004] Pulmonary drug delivery can directly deliver drugs to target organs, has the advantages of reducing drug dosage and reducing systemic toxic side effects, and becomes an ideal drug delivery route for the treatment of respiratory diseases. Dry powder inhalants, as an important form of pulmonary drug delivery, are preparations in which micronized drugs and carriers are in the form of capsules, bubbles or multi-dose reservoirs, which are inhaled by patients through special dry powder inhalation devices, and the inhaled drugs are delivered to the lungs. It is widely used in the treatment of diseases such as pneumonia and asthma.

[0005] However, the currently marketed dry powder inhalants have low lung effective deposition rate and low yield, which makes it difficult to ensure the effective drug content of the dose directly reaching the lung lesions. SUMMARY

[0006] In order to solve the technical problems of the above-mentioned existing anti-asthma dry powder inhalants, which have low lung effective deposition rate, resulting in difficulty in ensuring the effective drug content of the dose directly reaching the lung lesions. The present application provides a glutathione dry powder inhalant as well as a preparation method and application thereof. The glutathione dry powder inhalant is an inhalation preparation that is rapidly absorbed, convenient, safe and effective.

[0007] The present application uses the carrier and the dispersion enhancer to jointly maintain the particle dispersion, prevent glutathione from gathering, and serve as the "dry base" of the spray drying to provide the physical support for the preparation of the particles, adjust the particle size and the hygroscopicity, and ensure the effective deposition in the lung; meanwhile, the dispersion enhancer can enhance the dispersion capacity of the reduced glutathione, and ensure the uniform release of the dose during the inhalation. Through the addition of the carrier and the dispersion agent, the present application effectively improves the powder property of the reduced glutathione pulmonary dry powder inhalant, and makes the drug have good dissolution in the lung, improves the drug concentration in the lesion part, and improves the treatment effect of asthma. Moreover, the preparation of the preparation is simple, is beneficial to the production, and has high yield.

[0008] In the present application, the reduced glutathione, the carrier and the dispersion agent are mixed in the aqueous solvent system, and then are formed into the dry powder with the particle size of 0.5-5 μm through the spray drying, so as to obtain the glutathione dry powder inhalant.

[0009] The first object of the present application is to provide a glutathione dry powder inhalant, and the effective component of the glutathione dry powder inhalant is prepared from the following components in percentage by mass: 50-60% of the reduced glutathione, 10-20% of the carrier, and 20-30% of the dispersion agent, and the total is 100%.

[0010] Preferably, the carrier is mannitol, lactose or trehalose; and the dispersion agent is leucine.

[0011] The reduced glutathione can clear free radicals, protect the cell membrane, maintain the intracellular environment stable, and has potential therapeutic effect on the respiratory system diseases as an endogenous antioxidant and immunomodulator. The dry powder inhalant can directly deliver the drug to the lung, improve the drug concentration in the lung, and enhance the local treatment effect. The patients with respiratory system diseases often have the condition of increased oxidative stress, and the glutathione can reduce the oxidative damage in the lung, protect the lung tissue, and improve the lung function through the antioxidation. The present application takes the reduced glutathione as the medicinal active ingredient, and realizes the effective deposition in the lung through the dry powder inhalation, and improves the treatment effect of asthma.

[0012] More preferably, the carrier is mannitol, which is co-precipitated with the reduced glutathione during the spray drying process to form the composite particles with the suitable particle size and pore structure, so as to ensure the effective deposition in the lung during the inhalation. If the particle size is too large, it is easy to be deposited in the upper respiratory tract, and if the particle size is too small, it is easy to be exhaled with the airflow. Moreover, the mannitol has low hygroscopicity, which can reduce the hygroscopicity of the reduced glutathione, prevent the particle from caking during the storage, and maintain the powder fluidity. The crystallization characteristics of the mannitol can reduce the surface energy of the particles, reduce the interaction between the reduced glutathione molecules, avoid the aggregation of the active ingredient itself, and improve the mixing uniformity. In addition, the mannitol has the effect of dissolving mucus, has good physical and chemical stability, has no hygroscopicity, and can improve the aerosolization efficiency and stability of the drug.

[0013] Since the leucine molecule contains a hydrophobic isobutyl side chain, it can form a hydrophobic layer after adsorbing on the surface of particles, thereby reducing the electrostatic attraction and capillary force between particles. Therefore, the present application utilizes the hydrophobic leucine to modify reduced glutathione, utilizes the interfacial enrichment effect and crumpling forming characteristics of leucine to reduce particle aggregation and improve the flowability of the powder. At the same time, under the impact of the inhalation airflow, leucine can prevent secondary aggregation between particles, so that the composite particles are more easily dispersed into single particles, increase the fine particle fraction, and improve the lung deposition rate. Moreover, leucine is an essential amino acid for the human body, has low toxicity, and meets the stringent requirements of safety of auxiliary materials for inhalation preparations.

[0014] Preferably, the glutathione dry powder inhalation agent does not contain other pharmaceutical excipients, thereby avoiding the compatibility problems that may be caused by multiple excipients and improving the stability of the preparation. At the same time, the production steps are reduced, and the process is more easily scaled up and controlled through one-step granulation by spray drying. Moreover, the fewer the types of excipients, the lower the potential risk of irritation or allergy, and the preparation is particularly suitable for patients who need long-term inhalation therapy.

[0015] Preferably, the particle size of the glutathione dry powder inhalation agent is 0.5 μm to 5 μm, and the particle size range meets the requirement of 0.5 μm to 5 μm for pulmonary drug delivery, thereby ensuring that the drug particles can reach the alveolar region and improving the lung deposition rate and therapeutic effect.

[0016] Preferably, after the particle size treatment, the preparation can be directly filled into a dry powder inhaler in the form of a capsule or a vesicle.

[0017] A second object of the present application is to provide a preparation method of the above-mentioned glutathione dry powder inhalation agent, which comprises the following steps: Reduced glutathione, a carrier and a dispersing agent are dissolved in water to obtain a mixed solution.

[0018] The mixed solution is subjected to spray drying to obtain the glutathione dry powder inhalation agent.

[0019] Preferably, the spray drying conditions are as follows: the inlet temperature is 60°C to 120°C, the feeding rate is 1 mL / min to 10 mL / min, and the gas flow rate is 200 L / h to 1000 L / h. By adjusting the spray drying conditions, the particle size, morphology and component distribution of the glutathione dry powder inhalation agent are controlled, which meets the requirements of deposition in the lower respiratory tract and the lung, and the activity of glutathione is preserved during the spray drying process. At the same time, under the synergistic action of the carrier and the dispersing agent, low-density particles are formed, thereby reducing the inertial deposition.

[0020] Preferably, the solid content in the mixed solution is 0.5wt% to 2.5wt%. By controlling the solid content in the suspension, the solution viscosity and drying efficiency are balanced, thereby avoiding the clogging of the nozzle or the formation of excessively large particles.

[0021] Preferably, the inlet temperature and gas flow rate are maintained for 10-30 minutes after the spraying is completed, so as to completely remove the residual moisture, reduce the moisture absorption of the powder, and improve the long-term storage stability.

[0022] A third object of the present application is to provide use of the above-mentioned glutathione dry powder inhalant in the preparation of an asthma drug.

[0023] Compared with the prior art, the present application has the following technical effects: The present application effectively improves the properties of the glutathione pulmonary dry powder inhalant powder by adding the carrier and the dispersant, and the carrier and the dispersant are used to maintain the particle dispersity, prevent the aggregation of glutathione, adjust the particle size and moisture absorption, ensure the pulmonary deposition, enhance the dispersity of the particles, ensure the uniform release of the dose during inhalation, and in addition, the mannitol can also dissolve mucus and promote the deep lung delivery of the drug. Meanwhile, the present application adopts the spray drying method to accurately control the particle size distribution of the dry powder inhalant, so as to obtain the inhalant particles with a particle size of 0.5-5 microns, so that the drug is deposited in the bronchioles and alveolar regions of the lung to exert the drug efficacy; and the technical problem that the pulmonary effective deposition rate of most other commercially available dry powder inhalants is low, so that it is difficult to ensure the active ingredients of the dose reaching the lung lesion is solved.

[0024] The present application is proved by experiments that the glutathione pulmonary dry powder inhalant has the anti-asthma and anti-inflammatory efficacy. BRIEF DESCRIPTION OF DRAWINGS

[0025] Figure 1 The yield and water content of the glutathione dry powder prepared by adding different amino acid excipients.

[0026] Figure 2 The powder morphology of the glutathione dry powder prepared by adding different amino acid excipients after standing; wherein a is glutathione, b is L-threonine, c is L-isoleucine, d is L-leucine, and e is L-arginine.

[0027] Figure 3 The geometric particle size of the glutathione dry powder prepared by adding different amino acid excipients.

[0028] Figure 4 The yield and water content of the glutathione dry powder prepared by adding different sugar excipients and amino acids.

[0029] Figure 5 The scanning electron microscope image of the glutathione dry powder prepared by adding different sugar excipients and amino acids; wherein a is mannitol, b is lactose, and c is trehalose.

[0030] Figure 6 The geometric particle size of the glutathione dry powder prepared by adding different sugar excipients and amino acids.

[0031] Figure 7 Standard curve for reduced glutathione in HPLC assay.

[0032] Figure 8 SEM image of the glutathione dry powder inhaler prepared in Example 1.

[0033] Figure 9 Frequency distribution histogram of the particle size of the glutathione dry powder inhaler prepared in Example 1.

[0034] Figure 10 Settling rate distribution of the glutathione dry powder inhaler prepared in Example 1.

[0035] Figure 11 Settling rate distribution of the glutathione dry powder inhaler prepared in Example 2.

[0036] Figure 12 Settling rate distribution of the glutathione dry powder inhaler prepared in Example 3.

[0037] Figure 13 Schematic diagram of modeling mouse asthma model.

[0038] Figure 14 Neutrophil ratio of mouse blood after different drug treatments.

[0039] Figure 15 Total number of intermediate cells of mouse blood after different drug treatments.

[0040] Figure 16 Intermediate cell ratio of mouse blood after different drug treatments.

[0041] Figure 17 Total number of white blood cells of mouse blood after different drug treatments.

[0042] Figure 18 Tumor necrosis factor alpha concentration in mouse alveolar lavage fluid after different drug treatments.

[0043] Figure 19 Interleukin-1 beta concentration in mouse alveolar lavage fluid after different drug treatments. DETAILED DESCRIPTION

[0044] As described in the background art, the glutathione commonly used in the clinic at present is injection and tablet, and intravenous injection is often used to supplement glutathione in the body for treating asthma, but the aqueous solution of glutathione is unstable and cannot be stored for a long time, and injection can easily cause systemic reactions and has large adverse reactions, in addition, injection cannot be operated independently by patients, and can only be operated in hospitals.

[0045] Pulmonary administration has the advantages of reducing drug dosage and reducing systemic toxic side effects, because it can directly deliver drugs to the target organ, and becomes an ideal route for the treatment of respiratory diseases. Dry powder inhalation, as an important form of pulmonary administration, is a preparation in which micronized drugs and carriers are in the form of capsules, blisters or multi-dose reservoirs, and the atomized drugs are inhaled by patients through a special dry powder inhalation device to the lungs. It is widely used in the treatment of diseases such as pneumonia and asthma.

[0046] However, the current commercially available dry powder inhalation has a low lung effective deposition rate and a low yield, which makes it difficult to ensure the dose of active ingredients directly to the lung lesions.

[0047] Therefore, the present application prepares glutathione into a dry powder inhalation, which is convenient for long-term storage, ensures drug efficacy, allows patients to self-administer, and directly reaches the lungs. The inhalation has a rapid drug release, a quick onset of treatment, and less systemic adverse reactions. At the same time, the present application selects a glutathione dry powder inhalation with high yield, inhalation requirements and high lung deposition rate by screening the auxiliary materials and spray drying process parameters.

[0048] In order to enable those skilled in the art to better understand the technical solutions of the present application and to implement them, the present application will be further described below in conjunction with specific examples and drawings.

[0049] In the description of the present application, unless otherwise specified, the reagents used are commercially available, and the methods used are conventional techniques in the art.

[0050] Example 1 A glutathione dry powder inhalation.

[0051] The prescription 1 is as follows: Reduced glutathione 600mg; L-leucine 300mg; mannitol 100mg; pure water 40mL.

[0052] The specific preparation method is as follows: The prescription amount of reduced glutathione, L-leucine and mannitol is dissolved in 40mL of pure water, and then filtered with a 0.45μm filter to obtain a drug solution.

[0053] The drug solution is added to a spray dryer for spray drying, wherein the inlet temperature is set to 80℃, the feeding rate is 1mL / min, the fan power is 100%, and the gas flow rate is 800L / h. After the spray is completed, the gas inlet temperature and the gas flow rate are maintained for 30min to obtain a glutathione dry powder inhalation.

[0054] Example 2 A glutathione dry powder inhalation.

[0055] The prescription 2 is as follows: Reduced glutathione 500mg; L-leucine 300mg; mannitol 200mg; pure water 40mL.

[0056] The specific preparation method is as follows: The prescribed amounts of reduced glutathione, L-leucine, and mannitol were dissolved in 40 mL of pure water, and then filtered through a 0.45 μm filter membrane to obtain the drug solution.

[0057] The drug solution was added to a spray dryer for spray drying. The inlet temperature was set to 80℃, the feed rate to 1mL / min, the fan power to 100%, and the gas flow rate to 800L / h. After spraying, the gas inlet temperature and gas flow rate were maintained for 30 minutes to obtain glutathione dry powder inhaler.

[0058] Example 3 A glutathione dry powder inhaler.

[0059] Prescription 3 is as follows: Reduced glutathione 600mg; L-leucine 200mg; mannitol 200mg; pure water 40mL.

[0060] The specific preparation method is as follows: The prescribed amounts of reduced glutathione, L-leucine, and mannitol were dissolved in 40 mL of pure water, and then filtered through a 0.45 μm filter membrane to obtain the drug solution.

[0061] The drug solution was added to a spray dryer for spray drying, with the inlet temperature set at 80℃, the feed rate at 1mL / min, the fan power at 100%, and the gas flow rate at 800L / h. After spraying, the gas inlet temperature and gas flow rate were maintained for 30 minutes to obtain glutathione dry powder inhaler.

[0062] Example 4 A glutathione dry powder inhaler.

[0063] The prescription is as follows: Reduced glutathione 500mg; L-leucine 250mg; mannitol 250mg; pure water 40mL.

[0064] The specific preparation method is as follows: The prescribed amounts of reduced glutathione, L-leucine, and mannitol were dissolved in 40 mL of pure water, and then filtered through a 0.45 μm filter membrane to obtain the drug solution.

[0065] The drug solution was added into the spray dryer for spray drying, wherein the inlet temperature was set to 80°C, the feeding rate was 1 mL / min, the fan power was 100%, and the gas flow rate was 800 L / h; after the spray was completed, the gas inlet temperature and the gas flow rate were maintained for 30 min, to obtain the glutathione dry powder inhalant.

[0066] Example 5 A glutathione dry powder inhalant.

[0067] The prescription is as follows: Reduced glutathione 500 mg; L-leucine 250 mg; α-lactose 250 mg; pure water 40 mL.

[0068] The specific preparation method is as follows: The prescription amount of reduced glutathione, L-leucine, and α-lactose was dissolved in 40 mL of pure water, and then filtered using a 0.45 μm filter membrane, to obtain a drug solution.

[0069] The drug solution was added into the spray dryer for spray drying, wherein the inlet temperature was set to 80°C, the feeding rate was 1 mL / min, the fan power was 100%, and the gas flow rate was 800 L / h; after the spray was completed, the gas inlet temperature and the gas flow rate were maintained for 30 min, to obtain the glutathione dry powder inhalant.

[0070] Example 6 A glutathione dry powder inhalant.

[0071] The prescription is as follows: Reduced glutathione 500 mg; L-leucine 250 mg; trehalose 250 mg; pure water 40 mL.

[0072] The specific preparation method is as follows: The prescription amount of reduced glutathione, L-leucine, and trehalose was dissolved in 40 mL of pure water, and then filtered using a 0.45 μm filter membrane, to obtain a drug solution.

[0073] The drug solution was added into the spray dryer for spray drying, wherein the inlet temperature was set to 80°C, the feeding rate was 1 mL / min, the fan power was 100%, and the gas flow rate was 800 L / h; after the spray was completed, the gas inlet temperature and the gas flow rate were maintained for 30 min, to obtain the glutathione dry powder inhalant.

[0074] Comparative Example 1 A glutathione dry powder.

[0075] The prescription is as follows: Reduced glutathione 500 mg; L-arginine 500 mg; pure water 40 mL.

[0076] The specific preparation method is as follows: The prescribed amounts of reduced glutathione and L-arginine were dissolved in 40 mL of pure water, and then filtered through a 0.45 μm filter membrane to obtain the drug solution.

[0077] The drug solution was added to a spray dryer for spray drying. The inlet temperature was set to 80℃, the feed rate to 1mL / min, the fan power to 100%, and the gas flow rate to 800L / h. After spraying, the gas inlet temperature and gas flow rate were maintained for 30 minutes to obtain glutathione dry powder.

[0078] Comparative Example 2 A type of glutathione dry powder.

[0079] The difference from Comparative Example 1 is as follows: L-threonine was used.

[0080] Comparative Example 3 A type of glutathione dry powder.

[0081] The difference from Comparative Example 1 is as follows: L-Leucine was used.

[0082] Comparative Example 4 A type of glutathione dry powder.

[0083] The difference from Comparative Example 1 is as follows: L-Isoleucine was used.

[0084] This invention selects different amino acids as excipients to prepare glutathione dry powder inhalers and explores the effects of different amino acid excipients on the yield, moisture content and geometric particle size of the dry powder.

[0085] Depend on Figure 1 It can be seen that the yield of glutathione dry powder prepared by adding L-arginine, L-threonine, L-leucine or L-isoleucine to the reduced glutathione raw material did not change significantly; however, the moisture content changed significantly. Compared with the addition of L-arginine, L-threonine and L-isoleucine, the moisture content of glutathione dry powder after adding L-leucine was the lowest.

[0086] Depend on Figure 2 As can be seen from the actual images of glutathione dry powder, after adding different amino acid excipients and letting it stand for a period of time, the glutathione dry powder with added L-threonine changed color and hardened significantly, and agglomerated into lumps; the glutathione dry powder with added L-arginine also began to agglomerate into lumpy powder; while the powder with added L-isoleucine and L-leucine did not show obvious changes.

[0087] In this invention, the geometric particle size of dry powder containing L-leucine and L-isoleucine was measured and compared.

[0088] Depend on Figure 3 It can be seen that the particle size of glutathione powder with added L-isoleucine is significantly increased, while L-leucine shows no significant change compared to pure glutathione without amino acids, and the particle size meets the requirement of 0.5μm~5μm. Therefore, L-leucine was selected as the amino acid excipient for further experiments.

[0089] Sugars can alter the basic physicochemical properties of dry powder inhalers, and can also, together with amino acids, change the microstructure of the prepared dry powder inhaler particles. In this invention, mannitol, lactose, and trehalose were selected, and a glutathione dry powder inhaler was prepared according to a mass ratio of glutathione, L-leucine, and sugars of 1:0.5:0.5. The effects of different sugar excipients on the yield, moisture content, and morphology of the dry powder inhaler were investigated.

[0090] Depend on Figure 4 It can be seen that there is no significant difference in the dry powder yield when different sugar excipients are added; however, the glutathione dry powder inhaler with added mannitol has the lowest water content.

[0091] Depend on Figure 5 It is known that dry powder inhalers with added mannitol, trehalose, and lactose are basically spherical, but agglomeration occurs, which may be due to the fact that the spray drying parameters have not been optimized.

[0092] Depend on Figure 6 It is known that the dry powder inhaler with added mannitol has the smallest geometric particle size, the dry powder with added trehalose has the largest geometric particle size, and lactose is in between. This invention selects mannitol as a sugar excipient for further experiments.

[0093] Experimental test: 1. Liquid chromatography test.

[0094] This invention describes the liquid chromatography (LC) analysis of reduced glutathione raw material. The LC analysis conditions were established as follows: chromatographic column: Merck C18 column (4.6 mm × 250 mm, 5 μm); mobile phase: methanol and phosphate buffer solution (methanol:phosphate volume ratio 4:96); column temperature: 25 °C; flow rate: 1.0 mL / min; detection wavelength: 210 nm; injection volume: 10 μL. Linear regression was performed with peak area (A) as the ordinate and concentration (C) as the abscissa.

[0095] like Figure 7As shown in the figure, the standard curve equation for reducing glutathione is A=5938223.8806C+46507.9701, the correlation coefficient R2=0.9974, the accuracy, precision and recovery are good, which meet the requirements of the analysis method; and the linear range is 20 μg / mL-400 μg / mL.

[0096] 2. Scanning electron microscopy characterization.

[0097] From Figure 8 100 microspheres were randomly selected from the microspheres for particle size measurement, and a frequency histogram was prepared as shown in Figure 9 The average particle size of the glutathione dry powder inhalation is 1.96 μm, and the particle size range of the microspheres is 0.5 μm-4.5 μm, which meets the inhalation size requirement, and the particle size of the microspheres is mostly concentrated in 1.25 μm-3.5 μm, and the particles in this particle size range can be better deposited in the bronchioles and alveolar regions of the lung to exert the drug efficacy.

[0098] 3. In vitro inhalation property test.

[0099] The glutathione dry powder inhalation prepared in Example 1-Example 3 is respectively made into a drug-containing No. 3 capsule; wherein each capsule contains 50 mg of glutathione dry powder inhalation.

[0100] In this test, the Anderson eight-stage impactor (ACI) of the American TISCH company is used to investigate the in vitro lung deposition of the dry powder inhalation sample; the ACI device is used to simulate the human lung drug administration at a flow rate of 60 L / min, and the ACI device is composed of 8 collection trays, which are 1st-7th and microporous collection tray (MOC level). The spray holes are uniformly distributed above each collection tray, and the size of the spray holes decreases with the increase of the collection tray level. The dry powder in the capsule enters the ACI device along with the airflow to reach the 1st collection tray. The particles in the sample with an aerodynamic diameter greater than a certain value (related to the gas flow) will be trapped in the 1st collection tray, and the remaining particles will enter the next collection tray through the interstage channel. In this way, all particles in the dry powder inhalation are trapped on each collection tray in the order of aerodynamic diameter from large to small. The content of glutathione at each place can be determined by the following established glutathione HPLC determination method, and finally the distribution of the sample at each place is determined.

[0101] The full name of in vitro deposition rate is Fine Particle Fraction, abbreviated as FPF. FPF refers to the percentage of drug that can be deposited in the lungs out of the total amount of dry powder inhaler released from the device in a single dose. It is usually calculated based on the percentage of drug mass with an aerodynamic particle size between 0.5 μm and 5 μm out of the total collected drug mass. FPF is the most direct indicator for evaluating the pulmonary drug delivery efficiency and efficacy of dry powder inhalers. Generally, the higher the FPF, the higher the pulmonary drug delivery efficiency of the dry powder inhaler.

[0102] like Figures 10 to 12 As shown, Figure 12 The highest amount of drug was deposited in the lungs, indicating that the highest amount of reduced glutathione was able to reach the lungs to exert its therapeutic effect.

[0103] 4. In vivo pharmacodynamic testing.

[0104] This invention selects the glutathione dry powder inhaler prepared in Example 2 for dry powder administration; 10 mg of reduced glutathione raw material is dissolved in 1 mL of PBS solution to obtain a glutathione solution, which is administered via tail vein injection or pulmonary administration. ICR mice are divided into five groups, with 12 mice in each group. The first group is the normal group, without any treatment, denoted as Normal; the second, third, fourth, and fifth groups are as follows: Figure 13 Establish a 21-day asthma mouse model; such as Figure 13 As shown, the treatment of the asthma mouse model was as follows: Mice in groups 2, 3, 4, and 5 were sensitized by intraperitoneal injection of 0.2 ml of 10% OVA in PBS solution on days 1, 7, and 14, respectively. Subsequently, from day 15 to day 21, each mouse was nebulized with 1 mL of 1% OVA solution daily. During this period, mice in group 2 received no treatment and were designated as the asthma group. Mice in group 3 received nebulization, during which 2 mg of glutathione dry powder inhaler was administered to the lungs daily, with a glutathione content of 1 mg, designated as the glutathione dry powder inhalation group, or GSH-1. Mice in group 4 received glutathione solution via tail vein injection, with a glutathione content of 1 mg, designated as the glutathione solution intravenous injection group, or GSH-2. Mice in group 5 received glutathione solution via pulmonary administration, with a glutathione content of 1 mg, designated as the glutathione solution inhalation group, or GSH-3. Mice were sacrificed on day 22. Blood was collected after sacrifice for routine blood analysis and detection of inflammatory factors.

[0105] like Figure 14As shown, compared to the normal mice in the first group, the number of neutrophils in the blood of the mice in the asthma group was significantly increased, indicating that the asthma model was successfully established. Compared to the asthma group, the number of neutrophils in the blood of the mice in the other glutathione solution intravenous injection group, glutathione solution inhalation group, and glutathione dry powder inhalation group was decreased; among them, the decrease in the glutathione solution intravenous injection group was not significant; the glutathione dry powder inhalation group and the glutathione solution inhalation group both showed significant decreases, with a significant trend, indicating that glutathione inhalation administration has good efficacy.

[0106] like Figure 15 As shown, compared with the normal group of mice, the total number of eosinophils, basophils and monocytes in the blood of mice in the asthma group was significantly increased; the total number of intermediate cells in the blood of mice in the other glutathione solution intravenous injection group, glutathione solution inhalation group and glutathione dry powder inhalation group was significantly reduced, indicating that glutathione has good therapeutic effect.

[0107] like Figure 16 As shown, compared to the normal group of mice, the proportion of intermediate cells in the blood of mice in the asthma group was significantly increased, while the proportion of intermediate cells in the blood of mice in the other glutathione solution intravenous injection group, glutathione solution inhalation group, and glutathione dry powder inhalation group was significantly decreased. This indicates that glutathione, whether administered in solution or dry powder form, has a certain therapeutic effect on asthma; among them, the glutathione dry powder lung administration group showed the most significant effect, indicating that the glutathione dry powder inhalation group is more effective than the tail vein injection group in treating asthmatic mice.

[0108] like Figure 17 As shown, compared with the normal group of mice, the white blood cell count in the blood of the asthma group mice was significantly increased, while the white blood cell count in the blood of the other groups of mice in the intravenous glutathione solution group, the inhalation group of glutathione solution group, and the inhalation group of glutathione dry powder group was decreased; and the white blood cell count in the glutathione dry powder inhalation group and the glutathione solution inhalation group was significantly decreased, with the glutathione dry powder inhalation group showing the most significant effect, indicating a good therapeutic effect.

[0109] like Figure 18 As shown, compared with the normal group of mice, the tumor necrosis factor α (TNF-α) in the bronchoalveolar lavage fluid of the asthmatic mice was significantly increased, while the TNF-α in the bronchoalveolar lavage fluid of the other groups (intravenous glutathione solution, inhaled glutathione solution, and dry powder inhalation) was decreased. Moreover, the dry powder inhalation group and the intravenous glutathione solution administration group showed significant decreases, with the glutathione dry powder inhalation lung administration group showing the most significant decrease, indicating a good therapeutic effect.

[0110] like Figure 19As shown, compared with normal group mice, the interleukin-1β (IL-1β) in the bronchoalveolar lavage fluid of the asthma group mice is obviously increased, the IL-1β in the bronchoalveolar lavage fluid of the glutathione solution intravenous injection group, the glutathione solution inhalation group and the glutathione dry powder inhalation group mice is decreased; and the glutathione solution intravenous injection group and the glutathione dry powder administration group are significantly decreased, and the glutathione dry powder inhalation group is more significant.

[0111] Although preferred embodiments of the application have been described, those skilled in the art will recognize that additional modifications and variations may be made thereto without departing from the spirit and scope of the application. It is therefore intended that the appended claims cover all such modifications and variations as fall within the scope of the application.

[0112] Obviously, numerous modifications and variations of the present application are possible in light of the above teachings. It is therefore to be understood that within the scope of the appended claims and their equivalents, the application can be practiced otherwise than as specifically described.

Claims

1. A dry powder inhaler of glutathione, characterized by, The effective component of the glutathione dry powder inhalant is prepared from the following components in percentage by mass: 30-60% reduced glutathione, 10-20% carrier, 20-30% dispersing agent, and the total is 100%. The carrier is mannitol, lactose or trehalose, and the dispersing agent is leucine.

2. The dry powder inhaler of glutathione according to claim 1, characterized in that, The geometric particle size of the glutathione dry powder inhalant is 0.5-5 μm.

3. A method of preparing the dry powder inhaler of glutathione according to any one of claims 1 to 2, characterized in that, The method comprises the following steps: dissolving the reduced glutathione, carrier and dispersing agent in water to obtain a mixed solution; spray drying the mixed solution to obtain the glutathione dry powder inhalant.

4. The method of claim 3, wherein the dry powder inhaler of glutathione is prepared by, The solid content in the mixed solution is 0.5-2.5 wt%.

5. The method of claim 3, wherein the dry powder inhaler of glutathione is prepared by, The spray drying conditions are: inlet temperature of 60-120 °C, feed rate of 1-10 mL / min, and gas flow rate of 200-1000 L / h.

6. The method of claim 3, wherein the dry powder inhaler of glutathione is prepared by, After the spray drying is completed, the gas inlet temperature and gas flow rate are maintained for 10-30 min to obtain the glutathione dry powder inhalant.

7. Use of a dry powder inhaler of glutathione in the manufacture of a medicament for asthma, characterized in that, The glutathione dry powder inhalant is the glutathione dry powder inhalant according to any one of claims 1-2.

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