Fudosteine Nebulization Inhalation Solution Composition, Medicinal Assembly and Its Application

Abstract

There is provided a budesonide nebulization inhalation solution composition and its application. The composition, which is a budesonide nebulization inhalation solution composed of budesonide, a pH adjuster, and water, has, as the particle size of the spray droplets, D 90 The particle size is 7.0 to 10.0 μm, D 50 The particle size is 2.8 to 4.5 μm, D 10 The particle size is 0.5 to 1.5 μm, and the deposition percentage of particles with an aerodynamic diameter of 1.4 to 5.4 μm in the budesonide inhalation solution is 50% or more. Further provided is a medicinal assembly for use in combination with a nebulization inhalation solution, comprising an inhalation composition, a nebulizer with specific parameters, and a nebulizer cup. The budesonide nebulization inhalation solution composition and its medicinal assembly have the advantages of a small effective amount, rapid onset of effect, and high drug delivery efficiency, and are particularly effective in the treatment of tracheal and main bronchial diseases.

Description

Cross-reference

[0001] This application claims priority based on a Chinese patent application filed with the China National Intellectual Property Administration on July 18, 2022, with application number 202210842971.5 and title of invention "Fudosteine Nebulization Inhalation Solution Composition, Medicinal Assembly and Its Application", the entire content of which is incorporated herein by reference in its entirety.

Technical Field

[0002] The present invention relates to the technical field of nebulized inhalation, and particularly to a fudosteine nebulization inhalation solution composition, a medicinal assembly and its application.

Background Art

[0003] A nebulization inhalation solution composition refers to a preparation that administers a drug using a special device and exerts a systemic or local effect via the deep part of the respiratory tract, cavities, mucous membranes, etc. Compared with conventional drug delivery methods, nebulized drug delivery has the advantages of rapidity, high efficiency, and good patient compliance. The first-pass effect of the liver is avoided, and the side effects that may be caused by systemic drug delivery are reduced. Therefore, it is currently recognized worldwide as the best method for treating respiratory diseases such as asthma and chronic obstructive pulmonary disease (COPD). In recent years, as air pollution and aging progress, the prevalence of respiratory diseases has been increasing steadily, and nebulized inhalation administration has attracted attention.

[0004] As a specific dosage form of drug administration, a nebulized inhalant is converted from a nebulized inhalation solution into an aerosol through a specific device, and the inhaled dose is deposited in the respiratory system at a certain rate and with an appropriate particle size. In particular, the particle size has been widely studied by researchers as a factor of the preparation that mainly affects nebulized administration.

[0005] Fudosteine was first launched in Japan in 2001, manufactured by SSP and Mitsubishi Tanabe Pharma, belonging to the class of cysteine derivative drugs, which can change the composition and rheological properties of secretions, reduce the viscosity of sputum, and improve respiratory depression. Furthermore, fudosteine can increase serous secretion in the airway and suppress tracheal inflammation.

[0006] Currently, all fudosteine-related drugs sold at home and abroad are in oral dosage forms (including oral solutions, tablets, capsules, and granules). CN113975262A discloses a fudosteine-containing pharmaceutical composition containing fudosteine, thickening agents, flavoring agents, pH adjusters, coloring agents, fragrances, and solvents, and a method for producing the same. The pharmaceutical composition contains both a thickening agent and a pH adjuster, and the thickening agent and the pH adjuster are mutually formulated to maintain the excellent stability of the pharmaceutical composition. The pharmaceutical composition is an oral preparation, and auxiliary agents such as flavoring agents and fragrances are added to improve the taste. CN112057418A discloses a fudosteine oral solution containing fudosteine, metal ion compounds, acid-base regulators, and solvents, and a preparation method thereof. By forming a chelate compound between fudosteine and metal ions through chelation, the problem that the content of isomer impurities present in the fudosteine oral solution increases over time is solved. However, on the one hand, oral dosage forms are difficult to administer to patients with dysphagia or in a coma state. On the other hand, fudosteine can damage the gastric mucosal barrier and is likely to cause side effects such as rash, nausea, indigestion, itching, sensory numbness, abdominal discomfort, and diarrhea.

[0007] Chinese Patent CN108078964A discloses a fludosteine inhalation composition related to a fludosteine dry powder inhalant and its preparation method. Aiming at the disadvantages of commercially available fludosteine dosage forms, such as large dosage and insufficient systemic effects exerted, it aims to provide a fludosteine dry powder inhalant that can exert effects quickly at low doses and the drug can act directly on the lesion site, as well as its preparation method. However, since the dry powder inhalation device is relatively expensive, the cost increases, the burden on patients increases, and the internal flow resistance of the device affects the delivery and distribution of the drug, thus affecting the exertion of the drug effect.

[0008] US20200316003A1 first discloses a nebulized inhalation solution preparation of fludosteine. A nebulized inhalation solution preparation of a single dose of fludosteine containing fludosteine or its salt and / or its hydrate, the metal chelating agent EDTA, a pH adjuster, and water for injection was manufactured. The preparation has the characteristics of high efficiency, low toxicity, excellent stability, and high safety. However, when administered using a nebulizer, the problem that the particle size distribution of the spray droplets is wide, the range of the geometric standard deviation is large, the volume of the remaining chemical solution is large, the treatment time is extended, and the compliance is reduced has not been solved yet.

[0009] For conventional drugs, the action site administered through a pneumatic nebulizer is generally the lungs. However, fludosteine is mainly used as a mucolytic agent and can act well on goblet cells. However, since goblet cells are distributed in the trachea and main bronchi, it is necessary to solve the problem of the administration site of fludosteine in order to better the therapeutic effect of fludosteine. Specifically, it is necessary to enable fludosteine to mainly deposit in the trachea and main bronchi.

[0010] Therefore, the development of a nebulized inhalation solution composition of fludosteine and its pharmaceutical assembly that can solve the above problems has become an urgent problem to be solved.

Summary of the Invention

Problems to be Solved by the Invention

[0011] In order to solve the technical problem of the site of action after inhalation of budesonide by conventional spraying, the present invention provides a nebulized inhalation solution composition of budesonide having a uniform particle size distribution, high drug solution utilization rate, short treatment time, and remarkable treatment effect. In the present invention, furthermore, a medicinal assembly for the budesonide inhalation solution is provided. By using the budesonide inhalation solution in combination with a compression nebulizer, the budesonide spray particles can directly act on the trachea and main bronchi, achieving the most ideal treatment effect, significantly improving the safety and effectiveness in the administration to patients, and solving the technical problem that most of the conventional budesonide only deposits in the lungs.

Means for Solving the Problems

[0012] In order to achieve the above object, the following technical solutions are used in the present invention.

[0013] The first object of the present invention is to provide a nebulized inhalation solution composition of budesonide containing budesonide or a pharmaceutically acceptable salt thereof as an active ingredient, a pH adjuster, and water for injection.

[0014] Preferably, the pH of the composition is 2.5 to 5.0.

[0015] Preferably, the pH of the composition is 3.5 to 4.0.

[0016] More preferably, the pH of the composition is 3.7 to 3.9.

[0017] The aerodynamic particle size distribution of the spray particles is determined by the physical properties of the nebulized inhalation solution preparation, but the aerodynamic particle size of the spray particles and each parameter of the spraying device have an important influence on the administration site. The inventors of the present invention studied the main hyperplasia sites of goblet cells in the respiratory tract in the efficacy research and tests, and found that goblet cells were scattered and hyperplasia occurred in the tissues of the trachea and main bronchi of the respiratory tract, and that goblet cells mainly existed in the trachea and main bronchi. Fudosteine, an expectorant, inhibits the hyperplasia of goblet cells that secrete mucus in the trachea and reduces the viscosity of sputum to facilitate expectoration when used as an airway mucolytic agent, thus exerting an expectorant effect. Therefore, fudosteine acts on goblet cells and becomes excellent in expectorant effect. Thereby, when the administration site of fudosteine is the trachea and main bronchi, the expectorant effect is better. According to the pharmaceutical assembly of the fudosteine inhalation solution of the present invention, by using the fudosteine inhalation solution in combination with a compression nebulizer, the spray particles can directly act on the trachea and main bronchi, so that the most ideal therapeutic effect can be achieved.

[0018] Preferably, as the particle size of the spray droplets during inhalation of the inhalation solution by spraying, the D90 particle size is 7.0 to 10.0 μm, the D50 particle size is 2.8 to 4.5 μm, the D10 particle size is 0.5 to 1.5 μm, and the deposition percentage of particles with an aerodynamic diameter of 1.4 to 5.4 μm in the fudosteine inhalation solution is 50% or more.

[0019] Preferably, the particles with an aerodynamic diameter of 1.4 to 5.4 μm account for 53.2% or more by mass of the fudosteine inhalation solution, for example, 54.9%, 55.1%, 63.5%, 67.9%.

[0020] Preferably, the particles with an aerodynamic diameter of 2.1 to 5.4 μm account for 41.5% or more by mass of the fudosteine inhalation solution, for example, 44.6%, 46.7%, 49.4%, 50.3%.

[0021] Preferably, as the particle size of the spray droplets during inhalation of the inhalation solution by the spray, the D90 particle size is 7.0 to 9.5 μm, the D50 particle size is 2.8 to 4.4 μm, and the D10 particle size is 0.5 to 1.0 μm.

[0022] Preferably, as the particle size of the spray droplets during inhalation of the inhalation solution by the spray, the D90 particle size is 7.13 to 9.42 μm, the D50 particle size is 2.81 to 4.38 μm, and the D10 particle size is 0.54 to 0.95 μm.

[0023] Preferably, the concentration of the active ingredient in the composition is 20 to 140 mg / mL.

[0024] Preferably, the concentration of the active ingredient in the composition is 80 mg / mL.

[0025] Preferably, the pH adjuster is selected from the group consisting of hydrochloric acid, sulfuric acid, tartaric acid, lactic acid, citric acid, glacial acetic acid, malic acid, sodium citrate, sodium hydroxide, and combinations thereof.

[0026] Preferably, the nebulized inhalation composition has a driving gas flow rate of 4 to 10 L / min during use.

[0027] More preferably, the driving gas flow rate is 4 to 8 L / min.

[0028] Another object of the present invention is to provide an assembly of a fogging inhalation solution composition of fodosteine, comprising the fogging inhalation solution composition of fodosteine and a spraying device used in combination with the composition.

[0029] The spraying device includes a nebulizer and a cup for the nebulizer. Preferably, for the nebulizer, the pressure parameter of the compressor is 1.1 to 1.6 bar, the spraying speed is 0.4 to 0.6 ml / min, and the average particle size of the generated particles for a 0.9 wt% standard sodium chloride solution by the cup for the nebulizer is 2.5 to 3.5 μm.

[0030] More preferably, the nebulizer has a compressor pressure parameter of 1.2 to 1.6 bar and a spraying rate of 0.5 to 0.6 ml / min.

[0031] Another object of the present invention is to provide the use of budesonide aerosol inhalation solution composition, or an assembly containing budesonide aerosol inhalation solution composition, in the preparation of a drug for expectoration treatment of respiratory diseases. The use includes (1) a step of adding budesonide or a pharmaceutically acceptable salt thereof as an active ingredient to a solvent and dissolving it; (2) a step of adding a pH adjuster to the solution obtained in step (1) so as to adjust the pH of the solution to 3.7 to 3.9 to obtain an inhalation solution composition; (3) a step of filtering, dispensing, sealing, and packaging the inhalation solution composition; (4) a step of spraying the composition using a spraying device, and includes Preferably, the spraying device includes a nebulizer and a nebulizer cup. Preferably, the nebulizer has a compressor pressure parameter of 1.1 to 1.6 bar, a spraying rate of 0.4 to 0.6 ml / min, and an average particle size of generated particles with respect to 0.9 wt% standard sodium chloride solution by the nebulizer cup of 2.5 to 3.5 μm.

[0032] Preferably, the drug is administered to one or more administration sites selected from the group consisting of the nose, throat, trachea, esophagus, and main bronchus. More preferably, the drug is administered to the administration sites of the trachea and main bronchus.

[0033] Preferably, the budesonide drug is used for expectoration treatment for one or more selected from the group consisting of bronchial asthma, chronic asthmatic bronchitis, bronchiectasis, tuberculosis, silicosis, chronic obstructive pulmonary emphysema, atypical mycobacteriosis, pneumonia, and diffuse panbronchiolitis.

[0034] From the pharmacokinetic study, it was shown that after administration by spraying, the time to reach the maximum concentration in plasma and each tissue (lung, trachea, main bronchus, larynx, tongue, nose, spleen, brain) was shorter than that after oral administration, and it was more rapidly and widely distributed to each tissue in vivo compared to the case where fostein tablets were administered. In the case of spray administration, the maximum concentration in the trachea, main bronchus, esophagus, larynx, nose, and lung was significantly higher than that in the case of oral administration. In particular, the maximum concentration in the trachea and main bronchus was significantly higher than that in the case of oral administration, and the maximum concentration in plasma, liver, heart, spleen, and kidney was lower than that in the case of oral administration. Therefore, fostein administered by spraying has a higher maximum concentration in the tissues and organs of the respiratory system and a lower maximum concentration in the metabolic and excretory organs compared to fostein tablets administered orally. The inhaled solution composition of fostein administered by spraying has a shorter half-life in plasma, lung, main bronchus, liver, and brain compared to fostein tablets administered orally, and a longer half-life in the esophagus compared to fostein tablets administered orally. Fostein after administration accumulates little in plasma and tissues, and the concentration 24 hours after administration in tissues and plasma is lower than the lower limit of quantification, with a fast disappearance rate.

[0035] The atomized inhalation solution composition of fostein according to the present invention has a dosage of 2 to 5 mL. From the results of animal experiments, it was shown that the atomized inhalation solution composition of fostein has a significantly lower minimum effective dose than the commercially available fostein oral solution.

Advantages of the Invention

[0036] The beneficial effects of the present invention are as follows.

[0037] (1) In the present invention, for the characteristics of the treatment of tracheal and main bronchial diseases by budesonide, a nebulized inhalation solution composition of budesonide and a compression type spraying device (nebulizer, nebulizer cup) used in combination therewith are designed, the pressure parameters of the nebulizer and the output parameters of the nebulizer cup are limited and adjusted, and the particle size and particle size parameters (D10, D50, D90) of the spray droplets of the nebulized inhalation solution composition of budesonide are correspondingly controlled, so that the residual of the chemical solution is reduced, the drug delivery rate and the total delivery amount are improved, and the drug deposition percentage of the budesonide spray particles in the trachea and main bronchus is improved. That is, the budesonide spray particles can directly act on the trachea and main bronchus, and thus the most ideal expectorant effect is achieved.

[0038] (2) The nebulized inhalation solution composition of budesonide disclosed in the present invention has special physical properties. According to the composition of the present invention, the drawbacks such as wide spray droplet distribution, long treatment time, and poor compliance existing in the inhalation preparations of budesonide by the same type of spraying are overcome.

[0039] (3) In the present invention, by a reasonable component formulation that does not contain stabilizers such as EDTA in the drug formulation, the risk of side effects such as cough and asthma caused by drug administration is avoided.

[0040] (4) Compared with the commercially available oral preparations of budesonide of the same type, the nebulized inhalation solution composition of budesonide disclosed in the present invention has a smaller effective amount, a faster onset of effect, a smaller total dose during administration, and is also excellent in drug safety.

Brief Description of the Drawings

[0041]

Figure 1

Embodiments for Carrying Out the Invention

[0042] In order to further explain the object, technical solution, and beneficial effects of the present invention, the atomization inhalation solution composition, assembly, and their applications provided by the present invention will be described in more detail with specific embodiments. The described embodiments and examples are merely illustrative and do not represent all the technical solutions of the present application. Other technical aspects obtained by those skilled in the art without creative work based on the embodiments of the present invention are also included in the protection scope of the present invention.

[0043] I. Explanation of Terms As specific abbreviations used in the present invention, "FPD" is an abbreviation for fine particle dose, "MMAD" is an abbreviation for mass median aerodynamic diameter, GSD is an abbreviation for geometric standard deviation, and "NGI" is an abbreviation for Next Generation Impactor.

[0044] The term "particle size" means that which is measured for a single particle or particle distribution. These parameters can be measured by various techniques including dynamic light scattering, static light scattering, laser diffraction, sedimentation, time-of-flight, or other methods known to those skilled in the art. The particle size distribution can also be quantified from the size corresponding to the distribution (Dx) at a specific percentage point, provided that a certain percentage (x) of the whole is smaller than the defined size. For example, a D90 value distribution of 500 nm means that 90% (based on volume) of the distribution has a size less than 500 nm.

[0045] II. Experimental Conditions In the following examples, unless otherwise specified, the conditions of the experimental operations are as follows.

[0046] Chromatography Conditions Column: C18, 4.6×150 mm, 5 μm; Mobile phase: Phosphate solution (1→1000) of 5 mmol / L sodium heptanesulfonate - methanol (90:10) Detection wavelength: 210 nm; Column temperature: 50 °C; Injection volume: 10 μL; Flow rate: 1.0 mL / min; Solvent: Water

[0047] Preparation of calibration curve Preparation of stock solution of reference substance: Take 100 mg of the reference substance of fudosteine, accurately weigh it, put it into a 100 mL volumetric flask, dissolve and dilute it with the mobile phase, make up the volume to the calibration line, shake well, and use it as the stock solution. Preparation of calibration curve: Accurately pipette an appropriate amount of the stock solution, and serially dilute it with water to prepare standard solutions for calibration curve with concentrations of approximately 16 μg / ml, 40 μg / ml, 80 μg / ml, 96 μg / ml, and 160 μg / ml. Accurately take 10 μL of each of a series of solutions, inject them into the liquid chromatograph, perform linear regression analysis according to the least - squares method with the concentration on the horizontal axis and the peak area on the vertical axis, and obtain the linear regression equation: y = ax + b.

[0048] Method for measuring the dosage of fine particles The method was based on the method for measuring the aerodynamic characteristics of fine particles in the inhalation solution composition (General Chapter 0951, Part 4, Chinese Pharmacopoeia 2020 Edition) and NGI (Apparatus μsE) described in EP.

[0049] The specific operation is as follows. (1) To preheat the compressed - spray inhaler, place the NGI in a thermostat (or refrigerator) at 5 (±3) °C and hold for at least 90 min. (2) First, connect the NGI to the assembled leak detector, set the pressure of the leak detector to 4 KPa, then detect the leak rate. If the pressure becomes stable within 60 seconds, the test was conducted because the airtightness of the device is good. (3) Connect the nebulizer, NGI, large-capacity vacuum pump, and critical flow controller in order from left to right. Further, connect an air flow meter to detect the flow rate, turn on the large-capacity vacuum pump and the critical flow controller, and turn the adjustment knob to stabilize the magnitude of the flow rate. (4) Set the time parameter of the critical flow controller to 305 seconds. (5) Collect 2.5 mL of the sample, put it into the nebulizer cup, and at the same time turn on the "RUN" buttons of the compressed aerosol inhaler and the critical flow controller. Measure until 300 seconds, turn off the compressed aerosol inhaler, and turn off the large-capacity vacuum pump and the critical flow controller 5 seconds later.

[0050] (6) After all of the test process is completed, wash and transfer the parts of the nebulizer cup, artificial larynx + adapter, and collection trays (including filter trays) of stages 1 to 7 respectively. Accurately measure 20 ml of water, add it to the collection trays of Stages 1 to 6 respectively, mix uniformly, accurately measure 1 ml each and put it into a 10-ml volumetric flask, add water to make up the volume to the calibration mark for dilution, shake well, and obtain the test article solutions for Stages 1 to 6. Accurately measure 50 ml of water, add it to the collection tray of Stage 7, mix uniformly, and obtain the test article solution for Stage 7. Place the hierarchical filter paper of the MOC (Micro Orifice Collector) in a beaker, accurately measure 50 ml of water and put it into the beaker, squeeze the filter paper with a glass rod, stir uniformly, then filter, and take the secondary filtrate as the MOC test article solution. Take the artificial larynx, seal one end, accurately measure 50 ml of water and put it into the artificial larynx, shake well to obtain the artificial larynx test article solution, wash the nebulizer cup with an appropriate amount of water, transfer it into a 100-ml volumetric flask, add water to make up the volume to the calibration mark for dilution, shake well, then accurately transfer 1 ml to a 10-ml volumetric flask, add water to make up the volume to the calibration mark for dilution, shake well, and obtain the residual liquid of the nebulizer cup. Accurately measure 10 μL of each test sample solution respectively, inject them into the liquid chromatograph one by one, and record the chromatographic profile. Substitute the peak area A of each test sample into the linear equation y = ax + b to obtain the concentration c of each stage, and multiply by the dilution factor f of the sample solution to calculate the mass (m) of fudosteine at each stage. Calculation formula: Mass of fudosteine:

Number

[0051] Delivery rate and total delivery measurement method According to the "Method for Measuring Delivery Rate and Total Delivery Amount" in the section of inhalation liquid preparations (General Principles 0111, Volume IV, Chinese Pharmacopoeia 2020 Edition), the parameters of the breathing simulator were set according to the breathing characteristics.

[0052] For the test article solution, the parameters of the breathing simulator were set according to the breathing characteristics of adults. Exactly 2.5 mL of the sample was accurately measured and placed into the nebulizer cup. The breathing simulator, filter paper, nebulizer, and nebulizer cup were connected. At the same time, the nebulizer and the breathing simulator were turned on, and sprayed for 1 minute. The nebulizer was stopped once, and filter paper 1 was collected and placed in a 250 mL beaker. After replacing the new filter paper, spraying continued for 10 minutes, filter paper 2 was collected and placed in the second 250 mL beaker. Exactly 40 mL of water that was accurately measured was placed in the first beaker, and the filter paper was held below the liquid level while being fully wetted. After applying ultrasonic waves for 10 minutes, the filter paper was wrung out with a glass rod, stirred uniformly, and then filtered. Exactly 2 mL of the obtained secondary filtrate was accurately measured and placed into a 10 mL volumetric flask, diluted with water to the calibration line, shaken well, filtered, and the obtained secondary filtrate was collected. Exactly 50 mL of water that was accurately measured was placed in the first beaker, and the filter paper was held below the liquid level while being fully wetted. After applying ultrasonic waves for 10 minutes, the filter paper was wrung out with a glass rod, stirred uniformly, and then filtered. Exactly 1 mL of the secondary filtrate was accurately measured and placed into a 10 mL volumetric flask, diluted with water to the calibration line, shaken well, filtered, and the secondary filtrate was collected.

[0053] In the measurement method, each calibration curve solution and the test article solutions of filter paper 1 and filter paper 2 were accurately measured and injected into the liquid chromatograph respectively, and the chromatography profiles were recorded. The amount of budesonide collected on each filter paper was calculated by the calibration curve method based on the peak area. The ratio of the amount of budesonide collected on the first filter paper to the spraying time is the delivery rate, and the total amount of budesonide collected on each filter paper is the total delivery amount.

[0054] Unless otherwise specified, the formulation of the budesonide nebulized inhalation solution composition used in the examples was selected from the following Formulations 1 to 3. Prescription 1: Fudosteine (80 mg / mL), hydrochloric acid (appropriate amount), sodium hydroxide (added in appropriate amount as necessary), water for injection. Hydrochloric acid and sodium hydroxide were used to adjust the pH value. Prescription 2: Fudosteine (80 mg / mL), tartaric acid (appropriate amount), sodium hydroxide (added in appropriate amount as necessary), water for injection. Tartaric acid and sodium hydroxide were used to adjust the pH value. Prescription 3: Fudosteine (80 mg / mL), EDTA (0.5 mg / mL), hydrochloric acid (appropriate amount), sodium hydroxide (added in appropriate amount as necessary), water for injection. Hydrochloric acid and sodium hydroxide were used to adjust the pH value. Unless otherwise specified, Prescription 1 was selected as the prescription of the nebulized inhalation solution composition of fudosteine. Unless otherwise specified, the nebulized inhalation solution composition of fudosteine was nebulized under the nebulization conditions that the model number of the nebulizer was PARI BOY, the pressure of the nebulizer was 1.2 bar, the model number of the nebulizer cup was PARI LC SPRINT, and the spraying rate was 0.6 ml / min.

[0055] The nebulized inhalation solution compositions of fudosteine according to Prescriptions 1 and 2 were prepared by a method including the following steps. S1: Add 50% - 80% of water for injection to the device for preparing the drug solution, control the water temperature at 25 ± 10°C, introduce protective nitrogen gas into the water for injection, maintain it until the end of drug solution preparation, and keep the nitrogen gas in the drug solution preparation tank at positive pressure. After measuring until the residual oxygen was less than 2 mg / L, the next step was carried out. S2: Slowly add the solid drug according to the prescription and stir until it is completely dissolved. S3: Add the pH adjuster to adjust the pH, and after measuring the residual oxygen to be less than 2 mg / L, the next operation was carried out. S4: Add water for injection to make up the total volume and stir to mix uniformly. S5: Perform membrane filtration, dispense the solution into ampoules, and introduce nitrogen gas. The nebulized inhalation solution composition of fudosteine according to Prescription 3 was adjusted in the same manner as Prescriptions 1 and 2, except that the solution obtained by membrane filtration in Step S5 was dispensed into ampoules.

[0056] III. Verification Results Application Example 1: According to the preparation methods of Formulations 1 to 3 and the atomization inhalation solution composition of fostein, the atomization inhalation solution composition of fostein was prepared, the pH was adjusted to 3.7, and the results of measuring its impurities and formulation stability are shown in Tables 1 to 3.

[0057] Table 1: Stability of Formulation 1

Table 1

[0058] Table 2: Stability of Formulation 2

Table 2

[0059] Table 3: Stability of Formulation 3

Table 3

[0060] From the results of formulation preparation, by adopting measures such as expelling oxygen while filling nitrogen during the preparation of the chemical solution, filling nitrogen during dispensing, and filling nitrogen into the packaging bag during the preparation process, it can be guaranteed that the product is not oxidized, and it was found that it basically coincides with the stability of the formulation with EDTA addition. It was shown that the atomization inhalation solution composition of fostein obtained by the preparation process of the present invention has good stability.

[0061] Application Example 2: The influence of experimental conditions on the spraying effect when atomizing the budesonide inhalation solution composition under experimental conditions of different driving gas flow rates was detected. The inventor of the present invention conducted a large number of parallel experiments and found that when the pH of the budesonide inhalation solution composition was in the range of 2.5 to 5.0, there was no significant difference in the spraying effect at the flow rate of the same driving device. To further explain the present invention, for any budesonide inhalation solution composition in this application example, the pH was specifically set to 4.0. The drug distribution is shown in Figure 1, and the detection results are shown in Table 4.

[0062] Table 4 Influence of different driving gas flow rates of spray atomization on drug spraying results

Table 4

[0063] From the above results, the following was found. As the driving gas flow rate increased, the delivery rate, total delivery amount, and deposition percentage (particle size range 1.4 μm to 5.4 μm) increased correspondingly, while the median diameter MMAD decreased more and more. When the flow rate was 2 L / min, the delivery rate, total delivery amount, and aerosol deposition percentage were too small, and the deposition percentages of particles (particle size range 2.1 μm to 5.4 μm) that tended to deposit in the main bronchus and trachea and particles (particle size range 1.4 μm to 2.1 μm) that tended to deposit in the lungs were significantly lower than those in the case of 4 to 8 L / min. And, from the results of clinical trials, it was common for particles of 5 to 10 μm to deposit in the upper airway and particles of 1 to 5 μm to deposit in the lower airway. Therefore, since it is desired that the aerosol inhalation drug deposits in the lower airway, the median diameter MMAD exceeded 5 μm and did not meet the general requirements of aerosol inhalation. When the driving gas flow rate exceeded 4 L / min, the delivery rate, total delivery amount, aerosol deposition percentage (particle size range 1.4 μm to 5.4 μm), deposition percentage when the particle size range was 2.1 μm to 5.4 μm, and deposition percentage when the particle size range was 1.4 μm to 2.1 μm increased. In the case of Nos. 3 to 4 where the flow rate was in the range of 6 to 8 L / min, the increase in these indicators was not significant, but the general requirements for the delivery of the aerosol inhalation drug could be met.

[0064] Application Example 3: Nebulizers and nebulizer cups of different brands were selected, and the effective deposition dosage of the budesonide nebulized inhalation solution composition was investigated. The inventor of the present invention conducted a large number of parallel experiments and found that when the pH of the budesonide inhalation solution composition was within the range of 2.5 to 5.0, there was no significant difference in the effective deposition dosage when the same nebulizer and nebulizer cup were used. To further illustrate the present invention, for the budesonide inhalation solution composition in any of the present application examples, the pH was specifically set to 5.0, and the prescriptions of the samples were set as Prescription 1 and 2.

[0065] The atomization test of the samples was carried out, and the nebulizers and nebulizer cups used in the experiment were purchased as commercial products. The model numbers of the purchased nebulizers included PARI BOY SX, Yuwell 403E, Yuwell 403C, Yuwell 403M, Yinghua Rongtai FA, and Yinghua Rongtai HA03. The model numbers of the nebulizer cups included PARI LC SPRINT series blue core, PARI LC PLUS, and Yinghua Rongtai handheld blue respiratory valve. In Examples 1 to 5 and Comparative Examples 1 to 5 of the present application, the inhalation solution composition prescriptions of the present invention were used to investigate the influence of the parameters of different nebulizers and nebulizer cup assemblies on the spraying characteristics. The parameter conditions of Examples 1 to 5 are shown in Table 5, and the parameter conditions of Comparative Examples 1 to 5 are shown in Table 6.

[0066] Table 5 Prescriptions and Spraying Device Assemblies of Examples 1 to 5 of the Present Invention

Table 5

[0067] Table 6 Prescriptions and Spraying Parameters of Comparative Examples 1 to 5

Table 6

[0068] Table 7 Aerodynamic diameter and distribution by different nebulizers

Table 7

[0069] As shown in Table 7, D 50 has a certain linear relationship with MMAD, and it was found that as D 50 increased, MMAD also increased. In Examples 1 to 5, high deposition percentages (particle size range 1.4 μm to 5.4 μm), high delivery rates were shown, and the deposition percentages of particles (particle size range 2.1 μm to 5.4 μm) that tend to deposit in the trachea and main bronchi, and the deposition percentages of particles (particle size range 1.4 μm to 2.1 μm) that tend to deposit in the lungs all became high, and their MMAD values were less than 5 μm and relatively low. In contrast, in Comparative Examples 1, 2, and 5, the MMAD values exceeded 5 μm, making it difficult to form effective depositions in the trachea and lungs. In Comparative Examples 3 and 4, the MMAD was small, the particles mainly deposited in the alveoli, and it became difficult to exert a medicinal effect on the trachea.

[0070] From the above experimental results, by controlling the composition of the budesonide nebulized inhalation solution of the present invention in combination with the pressure of the compressor, the spraying speed, and the average particle diameter of the generated particles with respect to a 0.9 wt% standard sodium chloride solution by the cup for the nebulizer in the compression type spraying device used in combination therewith, it has become possible to control the particle size parameter of the sprayed droplets of the budesonide nebulized inhalation solution within a certain range. That is, as the particle size of the sprayed droplets, the D90 particle size is 7.0 to 10.0 μm, the D50 particle size is 2.8 to 4.5 μm, the D10 particle size is 0.5 to 1.5 μm, and the deposition percentage of the particles with an aerodynamic diameter of 2.1 to 5.4 μm in the budesonide inhalation solution can be controlled to be 41.5% or more. The drug delivery rate and the total delivery amount can be improved, and the sprayed particles can directly act on the trachea and the main bronchi, so that the most ideal expectoration effect is realized.

[0071] Application Example 4: The budesonide inhalation solution (Formulation 1) was administered to SD rats by single and continuous spraying, and the pharmacokinetic characteristics and distribution in its plasma and tissues were investigated. The tissue distribution characteristics of the budesonide inhalation solution were clarified by comparing with the tissue distribution of the budesonide tablets (original dosage form) administered orally once and continuously. The results are shown in Table 8.

[0072] Table 8 Comparison of the exposure amounts of budesonide in plasma and each tissue after single and continuous administration of a low dose of the budesonide inhalation solution and budesonide tablets (original dosage form) to SD rats

Table 8

[0073] As shown by the data in Table 8, oral administration of fosteamine resulted in high plasma exposure, while aerosol administration of fosteamine generally showed higher exposure in each tissue than in plasma, and the exposure to the respiratory system such as the larynx and trachea by aerosol administration was higher than that by oral administration. In the case of single aerosol administration, the exposure to the larynx, trachea, and esophagus was 1.34 to 3.19 times that of oral administration, and was lower in other tissues than in the case of oral administration. In the case of multiple aerosol administrations, the exposure (AUC0~t (hr*ng / g)) to the nose, larynx, trachea, esophagus, and main bronchi was 1.00 to 4.87 times that of oral administration. In particular, the exposure to the trachea and main bronchi was 3.43 and 4.87 times that of oral administration, respectively, and was lower in other tissues than in the case of oral administration. From these results, it was shown that according to the present invention, fosteamine can act well on the trachea and main bronchi.

[0074] The time to reach the maximum concentration of aerosol-administered fosteamine in plasma and each tissue (lung, trachea, main bronchus, larynx, tongue, nose, spleen, brain) was lower than that of oral administration, and it was distributed more rapidly and widely in each tissue in vivo compared to orally administered fosteamine tablets.

[0075] In the case of aerosol administration, the maximum concentration in the trachea, main bronchus, esophagus, larynx, nose, and lung was significantly higher than that of oral administration, and the maximum concentration in plasma, liver, heart, spleen, and kidney was lower than that of oral administration. Compared with orally administered fosteamine tablets, the fosteamine inhalation solution of the present invention had a higher maximum concentration in the tissues and organs of the respiratory system and a lower maximum concentration in the metabolic and excretory organs.

[0076] The aerosol-administered fosteamine inhalation solution had a shorter half-life in plasma, lung, main bronchus, liver, and brain than orally administered fosteamine tablets, a longer half-life in the esophagus than orally administered fosteamine tablets, and similar half-lives in other tissues (trachea, larynx, tongue, nose, heart, spleen, kidney).

[0077] The budesonide inhalation solution administered by continuous spraying and the budesonide tablets administered by continuous oral administration did not clearly accumulate in the plasma and tissues. The concentration of budesonide in the tissues and plasma at 24 hours after administration was lower than the lower limit of quantification, and the disappearance rate was rapid.

[0078] Application Example 5: Comparison of Pharmacokinetic Parameters of Oral and Inhaled Administration The budesonide inhalation solution (Formulation 1) was sprayed once on SD rats to investigate the pharmacokinetic characteristics of the budesonide inhalation solution in the rat body. Compared with the case of intravenous injection of the budesonide inhalation solution into SD rats, the absolute bioavailability of the budesonide inhalation solution (Formulation 1) in the rat body was investigated. Compared with the case of oral administration of the same amount of budesonide tablets to SD rats, the relative bioavailability of the budesonide inhalation solution (Formulation 1) in the rat body was investigated.

[0079] The results are shown in Table 9. Table 9 Collection of Average Pharmacokinetic Parameters of Different Dosage Forms of Budesonide

Table 9

[0080] Systemic exposure: When comparing the budesonide inhalation solution administered by spraying with the same dose of budesonide tablets administered orally, the budesonide inhalation solution had less exposure in the blood than the budesonide tablets. Bioavailability: When comparing the budesonide inhalation solution administered by spraying with the same dose of budesonide tablets administered orally, the relative bioavailability of the budesonide inhalation solution in the rat body was 16.2 ± 6.43%, the absolute bioavailability of the budesonide inhalation solution in the rat body was 18.0 ± 7.13%, the absolute bioavailability of the budesonide tablets in the flat body was 111 ± 22.6%, and the budesonide inhalation solution had lower bioavailability than the budesonide tablets. Time to reach the maximum concentration and maximum concentration: The budesonide inhalation solution was rapidly absorbed into the blood, with a shorter time to reach the maximum concentration and a lower maximum concentration in the blood compared to the budesonide tablets. Elimination half-life: There was no significant difference in the elimination half-life between the budesonide inhalation solution administered as a single spray and the same dose of budesonide tablets administered orally.

[0081] Application Example 6 Pharmacodynamic Test 1. Test article, control drug and reagents 1.1 Test article Budesonide inhalation solution (Formulation 1): 80 mg / mL, colorless and clear liquid, self-made.

[0082] 1.2 Control drug 1.2.1. Budesonide tablets were 0.2 g / tablet, white tablets, purchased from Jiangsu Zhengda Tianqing Pharmaceutical Co., Ltd., with batch numbers 2002151 and 2105141 respectively, and expiration dates of July 2021 and April 2023 respectively. Preparation: A certain amount of budesonide tablets were each taken and placed in a glassware and ground into fine powder, grinding while adding an appropriate amount of purified water, transferring to a graduated cylinder, adding ultrapure water and mixing well, making up the volume, and preparing solutions with concentrations of 68, 34, 17, 8.5, 4.25 mg / mL respectively for use in rat tests. Also, a solution with a concentration of 3.5 mg / mL was prepared for use in mouse tests. Note that fresh solutions were prepared and used immediately before use.

[0083] 1.2.2. Ambroxol hydrochloride oral liquid was 100 mL: 0.6 g, colorless to slightly yellowish clear liquid, purchased from China Resources Sanjiu (Nanchang) Pharmaceutical Co., Ltd., with batch number 2003004J and an expiration date until February 2022. Preparation: An appropriate amount of ambroxol hydrochloride oral liquid was weighed, and an appropriate amount of ultrapure water was added and mixed uniformly and well. A 4.5 mg / mL solution was prepared and used immediately before use.

[0084] 1.2.3. Prednisolone acetate tablets are white tablets at a dosage of 5 mg / tablet, purchased from Shanghai Shangyao Xinyi Pharmaceutical Factory Co., Ltd., with batch number 017200501 and an expiration date until May 2023. Preparation: A certain amount of prednisolone acetate tablets was taken and placed in a glassware, ground into fine powder, ground while adding an appropriate amount of purified water, transferred to a graduated cylinder, added with ultrapure water, mixed well, and made up to a constant volume. A solution with a concentration of 1.25 mg / mL was prepared immediately before use and used immediately.

[0085] 1.2.4. Acetylcysteine tablets: white tablets at a dosage of 0.2 g / tablet, purchased from ZAMBONS.p.A., with batch number 1002217 and an expiration date until February 2023. Preparation: A certain amount of acetylcysteine tablets was taken and placed in a glassware, ground into fine powder, ground while adding an appropriate amount of purified water, transferred to a graduated cylinder, added with ultrapure water, mixed well, and made up to a constant volume. A solution with a concentration of 43.2 mg / mL was prepared immediately before use and used immediately.

[0086] 1.2.5. Acetylcysteine solution for inhalation: colorless and clear liquid, purchased from ZAMBONS.p.A., with batch number 28005283 and an expiration date until May 2025.

[0087] 1.3. Reagents for model construction Lipopolysaccharide (LPS): white crystals, 100 mg per piece, purity over 99.5%, manufactured by SIGMA, with batch number 028M4094V and an expiration date until December 2023. Preparation: 100 mg of lipopolysaccharide was weighed, added with physiological saline and mixed uniformly, and made up to 100 mL. A lipopolysaccharide solution with a concentration of 1 mg / mL was prepared immediately before use and used immediately.

[0088] 1.4. Reagents 1.4.1. Physiological saline: colorless and transparent liquid, manufactured by Shijiazhuang No. 4 Pharmaceutical Co., Ltd., with batch number 2101031602 and an expiration date until November 2023.

[0089] 1.4.2. Sodium hydroxide: Sodium hydroxide with analytical grade purity, content of 96% or more, 500 g per bottle, purchased from Beijing Chemical Plant, batch number 20160912, expiration date until November 2022. Preparation: Weighed 20 g of sodium hydroxide, added ultrapure water and mixed uniformly, and made up to 500 mL. A solution with a concentration of 0.04 g / mL (1 mol / L) was prepared immediately before use and used right away. Preparation: Measured 1.8 mL of sodium hydroxide solution, added physiological saline and made up to 100 mL, mixed uniformly, and measured. The pH value was 11.9, that is, an alkaline sodium chloride solution was obtained.

[0090] 1.4.3. Phenol red: Phenol red with analytical grade purity, content of 99% or more, 25 g per bottle, manufactured by Shandong Xiya Chemical Co., Ltd., batch number B4301, expiration date until April 2022. Preparation: Weighed 2.5 g of phenol red, added alkaline sodium chloride solution and made up to 100 mL, adjusted with sodium hydroxide solution until the pH value reached 11.9, which was the same as the alkaline sodium chloride solution, and filtered through a 0.22 μm filter membrane. Preparation of phenol red calibration curve solution: Accurately weighed 50 mg of phenol red, added alkaline sodium chloride solution and made up to 50 mL, filtered through a 0.22 μm filter membrane to obtain a solution with a concentration of 1000 μg / mL. Diluted with alkaline sodium chloride solution to a total of 7 concentrations such as 10 μg / mL, 5 μg / mL, 2.5 μg / mL, 1.25 μg / mL, 0.63 μg / mL, 0.31 μg / mL, and 0.16 μg / mL.

[0091] 1.4.4. Sodium tetraborate (borax, Na2B4O7·H2O): Colorless hard crystalline powder, analytical grade purity, content of 99.5% or more, purchased from Tianjin Kaitong Chemical Reagent Co., Ltd., expiration date until September 2024. Preparation: A certain amount of Na2B4O7·H2O was weighed and dissolved in concentrated sulfuric acid to prepare a 25 mmol / L sodium tetraborate concentrated sulfuric acid solution.

[0092] 1.4.5. Carbazole (C 12 H9N): Colorless monoclinic flake crystals, analytical grade purity, purchased from Shanghai Yien Chemical Technology Co., Ltd., batch number RH297875, expiration date until September 2024. Preparation: 125 mg of C 12 H9N was weighed and dissolved in 100 mL of absolute ethanol to prepare a 7.5 mmol / L carbazole solution.

[0093] 1.4.6. Sodium chloride (NaCl): White cubic crystals or crystalline powder, analytical grade purity, content 99.5% or more, purchased from Tianjin Zhiyuan Chemical Reagent Co., Ltd., expiration date until April 2023. Preparation: 35.10 mg of pure NaCl was accurately weighed and dissolved in 200 mL of ultrapure water, mixed uniformly to prepare a sodium chloride solution with a concentration of 3 mol / L.

[0094] 1.4.7. D-Glucuronolactone (C6H8O6): White crystals or crystalline powder, analytical grade purity, content 99% or more, purchased from Shanghai Yien Chemical Technology Co., Ltd., batch number RH297877, expiration date until September 2024. Preparation: 50 mg of C6H8O6 was accurately weighed and dissolved in 50 mL of 3 mol / L sodium chloride to prepare a solution with a concentration of 1 mg / mL.

[0095] 2. Equipment Small animal nasal exposure system: Model HRH-MNE3026, manufactured by Beijing Huironghe Technology Co., Ltd. Microplate reader: Model Synergy HTX, manufactured by Bertin Instruments, USA. pH meter: Model PB~10, manufactured by Sartorius.

[0096] 3. Animals Mouse: SPF level of ICR strain; Rat: SPF level of SD strain. ICR mice, half male and half female, body weight 18 - 20 g, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., production license number SYXK(Beijing)2016 - 0006; SD rats, half male and half female, body weight 200 - 220 g, 220 - 240 g, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., production license number SCXK(Beijing)2016 - 0011.

[0097] 4. Test process 4.1. Data analysis method Route of administration: Administration by spray inhalation. The data generated from this test was analyzed using SPSS software, and a statistically significant difference between groups was shown as P < 0.05. Furthermore, the ED50 value was used to compare the effectiveness of Fudosteine tablets and Fudosteine inhalation solution.

[0098] 4.2. Consideration of spray generation conditions Administration was carried out using a spray device with a small animal nasal exposure system. After the aerosol concentration of the test article reached a steady state, particle size monitoring and analysis were performed. Two exposure ports were randomly selected, and each was measured twice, and the average value was obtained. Numerical values such as the mass median aerodynamic diameter (MMAD) and geometric standard deviation (GSD) of the aerosol particles were read out using a particle size distribution measuring device. A glass fiber filter membrane was placed at the randomly selected exposure port and measured under two spray generation conditions respectively. Samples were collected three times under each condition. After collecting the filter membrane, the concentration of the aerosol was detected using the LC - MS / MS method.

[0099] The results are shown in Table 10. Table 10 Results of consideration of spray generation conditions

Table 10

[0100] From the results, under the aerosol generation condition 1, the average aerosol concentration was 0.812 mg / L, the average particle size of the generated aerosol particles measured was 1.68 μm (corresponding to an average particle size of approximately 4.0 μm when measured with the Next Generation Impactor NGI), the D50 was 3.49 μm, and it was found that the sprayed drug met the requirements for the particle size to deposit in the airway and lungs and was helpful for exerting its drug effect. Under the aerosol generation condition 2, the average particle size became larger (corresponding to an average particle size of approximately 7.0 μm when measured with the Next Generation Impactor NGI). 4.3. Influence on the secretion of airway mucus

[0101] Based on the characteristic that phenol red is partially excreted from the airway after being intraperitoneally injected into mice, the excretion amount of phenol red in the trachea was measured to consider the influence of the test article on airway secretions. The higher the airway excretion amount, the stronger the expectorant effect of the Fudostein test article.

[0102] 4.3.1. Mouse phenol red excretion test ICR strain mice were randomly divided into 8 groups according to body weight, namely, the immediate model control group, the positive drug ambroxol hydrochloride group, the Fudostein tablet group (70.8 mg / kg), the high-dose group, medium-dose group, low-dose group of the Fudostein inhalation solution (14.6, 7.3, 3.65 mg / kg), and the comparison group - the high-dose group of the Fudostein inhalation solution. There were 14 animals in each group, with half being female and half being male. Each group of animals was administered once a day for 3 consecutive days according to the dosages, nebulizers, and routes shown in Table 11. Before the last administration, the animals in each group were fasted for 16 hours. 30 minutes after the last administration, 2.5% phenol red solution was intraperitoneally injected into each group of animals, and then 30 minutes later, the mice were sacrificed by cervical dislocation, the trachea was incised, 1.0 mL of alkaline sodium chloride solution was added, and it was vibrated with ultrasound for 5 minutes, and the excretion amount of phenol red in the airway was measured at a wavelength of 546 nm with a microplate reader.

[0103] Table 11 Influence on the phenol red excretion amount in mice(

Number

Table 11

[0104] As a result, in the positive drug ambroxol hydrochloride group and the high-dose group of fudosteine inhalation solution, the phenol red excretion amount into the trachea of mice was significantly increased compared with the model control group (P < 0.01; P < 0.05). In the fudosteine tablet group and the medium-dose and low-dose groups of fudosteine inhalation solution, the phenol red excretion amount into the trachea of mice tended to increase, but no statistical difference was observed. In the high-dose group of fudosteine inhalation solution in the comparison group with large spray particle diameters, the phenol red excretion amount into the trachea of mice tended to increase, but no statistical difference was observed.

[0105] Also, it was found that the spraying conditions also affect the drug efficacy results. Under spraying condition 2, when the dose was 15.3 mg / kg, the drug efficacy was clearly inferior compared with that when the dose was 14.6 mg / kg under spraying condition 1, and there was no significant difference in the drug efficacy in the medium-dose and low-dose groups compared with the model control group. Therefore, it was suggested that when spraying drugs on animals, it is necessary to strictly meet the spraying conditions.

[0106] 4.3.2. Detection test of airway secretion volume and mucopolysaccharide content in LPS-induced mucus hypersecretion rats SD rats were randomly divided into 11 groups according to body weight, namely, the aerosol blank control group of healthy animals, the acetylcysteine solution group for inhalation of positive drug, the high-dose group, medium-dose group, and low-dose group (8.75, 4.5, 2.25 mg / kg) of fudosteine inhalation solution, the aerosol blank control group of model animals, the fudosteine tablet group (42.5 mg / kg), the acetylcysteine solution group for inhalation of positive drug, the high-dose group, medium-dose group, and low-dose group (8.75, 4.5, 2.25 mg / kg) of fudosteine inhalation solution, with 10 rats in each group and half males and half females. All animals in the model group were made to inhale LPS solution (1 mg / mL, gas generation volume 10 L / min, dilution gas volume 1 L / min, 40 min / time / day) by spraying once a day for 2 consecutive days for modeling. Each group was administered once a day for 3 consecutive days according to the dosages and administration routes shown in Table 12. Immediately after the last administration, chloral hydrate was intraperitoneally injected into the rats for anesthesia, and they were fixed in the supine position. The skin of the neck was incised, the trachea was separated, and a glass capillary of known weight was inserted between the two cartilages at the lower end of the thyroid gland to aspirate sputum. When the capillary was filled with sputum, it was exchanged with another capillary. Using the increased weight of the capillary as the detection index, tracheal secretion drainage was collected and weighed to calculate the tracheal secretion volume in 2 hours (secretion volume = increased weight of capillary / body weight × 100 g).

[0107] After the drainage was completed, the trachea was removed from the larynx to the tracheal bifurcation, weighed, the trachea was cut longitudinally, a cotton swab was used to take out airway mucus, washed with 3 mL of distilled water, centrifuged at 3000 r / 15 min, and 2.5 mL of the supernatant was collected. The carbazole sulfate colorimetric method was used to measure the content of mucopolysaccharide in the trachea per unit weight of rats in each group at a wavelength of 530 nm.

[0108] During the processes of acute and chronic airway diseases, the degree of glycosylation increases significantly with the increase in the secretion of mucus and mucin, which is closely related to the severity of infection. Mucin is the main component of mucus and determines the adhesiveness, viscosity, and elasticity of airway mucus. The glycose at the end of mucin is collectively called mucopolysaccharide, and its main components are sialic acid and fucose. Therefore, in this study, the mucus repair effect of fudosteine inhalation solution was evaluated using the content of mucopolysaccharide as an index. In addition, the content of mucopolysaccharide and the secretion volume of airway mucus were measured in the study.

[0109] In view of the characteristic that the test article in this study is an inhalation preparation, after administering the test article to animals via the oral and nasal administration routes, a certain amount of inhalation solution may accumulate in the airways of the animals, increasing the secretion fluid in the airways of the animals. Also, administering physiological saline as a control to the blank control group and the model control group may still increase the secretion fluid. Since all previous airway secretion tests were conducted on normal animals, it is difficult to objectively evaluate the effectiveness of the inhalant on airway secretion. Therefore, in this test, it is important to observe and compare the effects of the test article on the mucus secretion volume of animals, and further evaluate the effects on the mucopolysaccharide content in airway mucus. Thus, a normal control group under physiological conditions and a mucus hypersecretion model control under pathological conditions were designed.

[0110] The results are shown in Table 12. Table 12 Effects on the secretion fluid volume and mucopolysaccharide content in the airways of LPS-induced mucus hypersecretion rats

Table 12

[0111] As a result, compared with the aerosol blank control group of healthy animals, in the high-dose and medium-dose groups of budesonide inhalation solution, the amount of airway secretion in rats increased significantly (P<0.01; P<0.05), and there was no significant difference at low dose. However, in the aerosol blank control group of model animals, the amount of airway secretion in rats increased significantly (P<0.01), suggesting that the modeling was successful. Also, in the model animal budesonide tablet group, the acetylcysteine solution group for inhalation of positive drug, and the high-dose and medium-dose groups of budesonide inhalation solution, compared with the aerosol blank control group of model animals, the amount of airway secretion in rats all increased significantly (P<0.01; P<0.05), and there was no significant difference at low dose.

[0112] Compared with the aerosol blank control group of healthy animals, in the acetylcysteine solution group for inhalation of positive drug, the high-dose, medium-dose, and low-dose groups of budesonide inhalation solution, the mucopolysaccharide content in the airway secretion of rats all increased significantly (P<0.01; P<0.05). However, in the aerosol blank control group of model animals, the mucopolysaccharide content in the airway secretion of rats increased significantly (P<0.01), suggesting that the modeling was successful. Compared with the aerosol blank control group of model animals, in the model animal budesonide tablet group, the acetylcysteine solution group for inhalation of positive drug, and the high-dose and medium-dose groups of budesonide inhalation solution, the mucopolysaccharide content in the airway secretion of rats all decreased (P<0.01; P<0.05), and there was no significant difference at low dose.

[0113] The test results showed that when the test article was administered by inhalation to normal rats, both the airway mucus secretion amount and the mucopolysaccharide content were significantly improved. However, in the comparative test between normal rats and mucus hypersecretion model rats, it was found that the content of mucopolysaccharide was significantly reduced by the test article. It was suggested that the test article played a role in inhibiting the excessive secretion of mucus by inhibiting the secretion of mucopolysaccharide in the airway.

[0114] 4.4. Influence on airway inflammation SD rats were randomly divided into 9 groups according to body weight, namely, blank control group, model control group, positive drug prednisolone acetate group, high-dose, medium-dose, and low-dose groups of fodosteine tablets (42.5 mg / kg, 21 mg / kg, 10 mg / kg), high-dose, medium-dose, and low-dose groups of fodosteine inhalation solution (8.75 mg / kg, 4.5 mg / kg, 2.25 mg / kg), with 8 rats in each group, half male and half female. LPS solution (1 mg / mL, generated gas volume 8 L / min, diluted gas volume 4 L / min, 40 min / time / day) was inhaled by spraying into the animals of each group except the blank control group, and modeling was performed once a day for 3 consecutive days. After modeling for 1 hour every day, the animals in each group were administered according to the doses and routes shown in Table 13. Four hours after the last modeling, the trachea and lungs were excised, fixed with 10% formalin, dehydrated, embedded, sliced, stained with HE, the tissue was examined pathologically, the number of neutrophils in the tissue was counted, and the differences between groups were compared.

[0115] The results are shown in Table 13. Table 13 Effects on LPS-induced airway inflammation in rats (

Number

Table 13

[0116] From the above results, compared with the blank control group, in the airways of the rats in the model control group, inflammatory cells mainly composed of neutrophils infiltrated, and the number of neutrophils increased significantly (P < 0.01). Compared with the model control group, in the positive drug prednisolone acetate group, the high-dose group of fodosteine tablets (42.5 mg / kg), the high-dose and medium-dose groups of fodosteine inhalation solution (8.75 mg / kg, 4.5 mg / kg), the quantity of neutrophils all decreased significantly (P < 0.01; P < 0.05). In the medium-dose and low-dose groups of fodosteine tablets and the low-dose group of fodosteine inhalation solution, the number of neutrophils decreased, but no statistical difference was observed.

[0117] In the above test, for the test article Fudosteine inhalation solution, after examining the inhibitory effect on the hyperplasia of goblet cells in the epithelial tissue of the main bronchus, the inhibitory effect on airway mucus secretion and airway inflammation, its expectorant effect was comprehensively evaluated. In the goblet cell hyperplasia model in the epithelial tissue of the main bronchus of LPS-induced rats, the effective doses of Fudosteine inhalation solution and the control drug Fudosteine tablets were compared, and the ED 50 values were calculated. As a result, for the Fudosteine inhalation solution, the minimum effective dose was 4.5 mg / kg and the ED 50 value was 3.38 mg / kg; for the Fudosteine tablets, the minimum effective dose was 42.5 mg / kg and the ED 50 value was 37.67 mg / kg. The difference in the minimum effective doses of both was 9.4-fold, and the difference in the ED 50 values was 11.1-fold.

[0118] Application Example 7: Consideration of the Expectorant Effect and Its Mechanism of Fudosteine Inhalation Solution The Fudosteine oral solution and the Fudosteine inhalation solution were respectively administered to LPS-induced sputum production model rats, and their expectorant effects were compared.

[0119] 1. Test Materials 1.1. Test Articles and Control Articles Fudosteine inhalation solution (Formulation 1): Colorless and clear liquid, 80 mg / mL, self-made. Fudosteine tablets are 0.2 g / tablet, white tablets, purchased from Jiangsu Zhengda Tianqing Pharmaceutical Co., Ltd., with batch numbers 2002151 and 2105141 respectively, and expiration dates of July 2021 and April 2023 respectively.

[0120] 1.2. Reagents Lipopolysaccharide (LPS) is a white crystal, 100 mg / stick, with a purity of over 99.5%, batch number 028M4094V, and was purchased from SIGMA in May 2021. Preparation method: Weigh a certain amount of LPS accurately and mix it with physiological saline to prepare an LPS solution with a concentration of 2 mg / mL. Physiological saline: Colorless and transparent liquid, batch number 1903111602.

[0121] 1.3. Equipment Analytical balance: CPA225D, manufactured by Sartorius. Automatic staining device: DRS 2000JC-D2, manufactured by SAKURA, Japan. Microscope: BX43, manufactured by OLYMPUS, Japan.

[0122] 1.4. Animals SD rats, a total of 96 (half male and half female), weighing 200 - 220 g, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd., production license number SCXK (Jing) 2016 - 0006.

[0123] 2. Test procedure An LPS solution was administered to rats by aerosol inhalation to induce the proliferation of goblet cells in their tracheas and main bronchi, and a rat sputum production model was established. The expectorant effect of the test article was evaluated by measuring the number of goblet cells in the tracheas and main bronchi of the animals.

[0124] Test method: The rats that passed the quarantine were randomly divided into a blank group, a model group, an aerosol high-dose group, an aerosol medium-dose group, an aerosol low-dose group, an oral high-dose group, an oral medium-dose group, and an oral low-dose group according to their body weights, with 8 rats in each group (half male and half female). Animals in each group other than the blank control group were administered an LPS solution with a concentration of 2 mg / mL by aerosol inhalation at a rate of 10 L / min of generated gas and 1 L / min of dilution gas for 40 minutes per time, twice a day, to establish a rat sputum production model.

[0125] On the day following the second modeling, purified water was administered to the blank group and the model group by forced oral administration, and the corresponding test article and control product at the indicated dosages were administered to the other groups by nebulized inhalation and oral administration at the dosages shown in Table 12, once a day for 5 consecutive days. Four hours after the last administration, the right lung was excised and subjected to AB-PAS staining. Sections of three lung bronchi were prepared from each animal, and three fields of view in each section were randomly selected and examined under a microscope. The length of the basement membrane and the number of goblet cells in each field of view were measured, and the number of goblet cells contained on the basement membrane per unit length (mm) was calculated. The average result from nine fields of view was taken as the measurement result of the goblet cell count for that animal.

[0126] 3. Test Results Compared with the blank group, the number of goblet cells in the lung bronchi of animals in the model group increased significantly (P < 0.01). Compared with the model group, the number of goblet cells in the trachea and main bronchi of animals in the high-dose nebulized group, medium-dose nebulized group, and low-dose nebulized group decreased significantly (P < 0.01), but there were no significant changes in the high-dose oral group, medium-dose oral group, and low-dose oral group (P > 0.05). Details are shown in Table 14.

[0127] Table 14 Effects on Goblet Cells in the Trachea and Main Bronchi of Sputum Generation Model Rats (

Number

Table 14

[0128] From the experimental results, compared with the blank control group, in the model control group, the number of goblet cells in the epithelial tissue of the main bronchus of rats increased significantly. Compared with the model control group, in the oral high-dose group, and the high- and medium-dose aerosol groups, the number of goblet cells in the epithelial tissue of the main bronchus of rats all decreased significantly, indicating that they had a significant expectorant effect (ability to resist goblet cell hyperplasia). In the oral medium-dose group, low-dose group, and low-dose aerosol group, the number of goblet cells in the epithelial tissue of the main bronchus of rats showed a decreasing trend, but there was no statistical difference. Since the minimum effective dose of Fudosteine inhalation solution in the sputum production model was 4.25 mg / kg, and the effective dose was much lower than 42.5 mg / kg of the control product (commercially available Fudosteine tablets), it was revealed that the nebulized inhalation solution composition of the present invention and its pharmaceutical assembly showed good therapeutic effects.

[0129] The above description of the embodiments is only used to assist in understanding the method of the present invention and its central concept. It should be noted that those skilled in the art can make some improvements and modifications to the present invention without departing from the principle of the present invention, and these improvements and modifications are also included within the protection scope of the claims.

Claims

1. A nebulized inhalation solution composition of fostein, comprising fostein or a pharmaceutically acceptable salt thereof as an active ingredient, a pH adjuster, and water for injection, wherein the pH is 2.5 to 5.0, when the inhalation solution is inhaled by spraying, as the particle size of the spray droplets, the D90 particle size is 7.0 to 10.0 μm, the D50 particle size is 2.8 to 4.5 μm, the D10 particle size is 0.5 to 1.5 μm, and the deposition percentage of particles with an aerodynamic diameter of 1.4 to 5.4 μm in the inhalation solution of fostein is 50% or more.

2. The deposition percentage of particles with an aerodynamic diameter of 1.4 to 5.4 μm in the fostein inhalation solution is 53% or more, and preferably, the deposition percentage of particles with an aerodynamic diameter of 2.1 to 5.4 μm in the fostein inhalation solution is 41.5% or more. The inhalation solution composition according to Claim 1, characterized in that.

3. When the inhalation solution is inhaled by spraying, as the particle size of the spray droplets, the D90 particle size is 7.0 to 9.5 μm, the D50 particle size is 2.8 to 4.4 μm, the D10 particle size is 0.5 to 1.0 μm, and preferably, when the inhalation solution is inhaled by spraying, as the particle size of the spray droplets, the D90 particle size is 7.13 to 9.42 μm, the D50 particle size is 2.81 to 4.38 μm, the D10 particle size is 0.54 to 0.95 μm. The inhalation solution composition according to Claim 1, characterized in that.

4. The concentration of the active ingredient in the composition is 20 to 140 mg / mL. The inhalation solution composition according to any one of Claims 1 to 3, characterized in that.

5. The concentration of the active ingredient in the composition is 80 mg / mL. The inhalation solution composition according to Claim 4, characterized in that.

6. The pH of the composition is 3.5 to 4.

0. The inhalation solution composition according to any one of Claims 1 to 3, characterized in that.

7. The pH of the composition is 3.7 to 3.

9. The inhalation solution composition according to Claim 4, characterized in that.

8. The pH adjuster is selected from the group consisting of hydrochloric acid, sulfuric acid, tartaric acid, lactic acid, citric acid, glacial acetic acid, malic acid, sodium citrate, sodium hydroxide, and combinations thereof. The inhalation solution composition according to any one of Claims 1 to 3, characterized in that.

9. An assembly comprising the nebulized inhalation solution composition of fostein according to any one of Claims 1 to 8, (1) An atomized inhalable solution composition according to any one of claims 1 to 8, (2) An assembly comprising a nebulizer having a compressor pressure parameter of 1.1 to 1.6 bar and a spray rate of 0.4 to 0.6 ml / min, and a nebulizer cup having an average particle size of generated particles of 2.5 to 3.5 μm in a 0.9 wt % standard sodium chloride solution, and a spray device to be used in combination with the composition.

10. 10. The assembly according to claim 9, characterized in that the nebulizer has a compressor pressure parameter of 1.2-1.6 bar and a nebulization rate of 0.5-0.6 ml / min.

11. Use of the fudosteine atomized inhalation solution composition according to any one of claims 1 to 8 or the assembly according to any one of claims 9 to 10 in the preparation of a drug for expectorant treatment of respiratory diseases, comprising: (1) adding and dissolving fudosteine or a pharma- ceutically acceptable salt thereof as an active ingredient in water for injection; (2) adding a pH adjuster to the solution obtained from step (1) so as to adjust the pH of the solution to 3.7-3.9 to obtain an inhalation solution composition; (3) filtering, dispensing, sealing and packaging the inhalation solution composition; (4) spraying the composition using a spray device; The spray device is characterized in that it comprises a nebulizer having a compressor pressure parameter of 1.1 to 1.6 bar and a spray rate of 0.4 to 0.6 ml / min, and a nebulizer cup having an average particle size of generated particles of 2.5 to 3.5 μm for a 0.9 wt% standard sodium chloride solution.

12. 12. The use according to claim 11, characterized in that the medicament is administered to one or more administration sites selected from the group consisting of the nose, throat, trachea, esophagus, and main bronchi.

13. The use according to claim 11, characterized in that the drug is used for expectorant treatment for one or more diseases selected from the group consisting of bronchial asthma, chronic asthmatic bronchitis, bronchiectasis, tuberculosis, pneumoconiosis, chronic obstructive pulmonary emphysema, atypical mycobacteriosis, pneumonia, and diffuse panbronchiolitis.

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