A bronchodilator salbutamol aerosol and a preparation method thereof

Abstract

The invention relates to a bronchodilator salbutamol aerosol and a preparation method thereof, and belongs to the technical field of drugs for treating respiratory diseases. The salbutamol aerosol of the invention is composed of salbutamol, triolein, ethanol, meglumine, and a propellant. The invention uses triolein to encapsulate salbutamol to prevent drug degradation and inactivation, thereby improving the local therapeutic effect of salbutamol in the respiratory tract, and also adds an appropriate amount of meglumine to the prescription of the salbutamol aerosol, thereby significantly improving the stability of the salbutamol aerosol, effectively reducing the content of related substances, and ensuring the safety of medication.

Description

Technical Field

[0001] The invention belongs to the technical field of drugs for treating respiratory diseases, and particularly relates to a bronchodilator salbutamol aerosol and a preparation method thereof. Background Art

[0002] The information disclosed in this background technology section is only intended to enhance the understanding of the overall background of the invention, and should not necessarily be regarded as an admission or any form of suggestion that the information constitutes the prior art already known to a person skilled in the art.

[0003] Respiratory diseases are common and frequently occurring diseases. The main lesions are in the trachea, bronchi, lungs and chest cavity. Patients with mild lesions often have coughs, chest pains and breathing problems. Patients with severe lesions have difficulty breathing, hypoxia and even respiratory failure leading to death. The mortality rate in cities ranks third, while it ranks first in rural areas. What should be paid more attention to is that due to air pollution, smoking, an aging population and other factors, the incidence and mortality rates of chronic obstructive pulmonary disease (COPD, including chronic bronchitis, emphysema, cor pulmonale), bronchial asthma, lung cancer, diffuse interstitial pulmonary fibrosis, and lung infections at home and abroad have increased.

[0004] Bronchial asthma is a common chronic inflammatory airway disease, characterized by recurrent wheezing, shortness of breath, chest tightness or coughing, which often occur or worsen at night or in the early morning. In severe cases, dyspnea and hypoxemia may occur. The onset of asthma is related to genetic and environmental factors. Genetic factors mainly determine the patient's prone constitution, while environmental factors such as various allergens, air quality, smoking, exercise, etc. are specific triggering factors. Common types of bronchial asthma include exercise-induced, drug-induced, occupational and allergic.

[0005] Asthma cannot be completely cured at present, but the condition can be controlled through medication to improve the quality of life. Commonly used medications include control medications and reliever medications. The former are used to control the condition in the long term, such as inhaled corticosteroids; the latter are used to relieve acute attacks, such as short-acting β 2 Receptor agonists, etc.

[0006] Salbutamol is a short-acting beta 2 Adrenergic receptor agonist, chemical formula C 13 H 21 NO 3 , white crystalline powder, odorless, soluble in ethanol, slightly soluble in water, used as an antiasthmatic, can effectively inhibit the release of allergenic substances such as histamine, prevent bronchospasm, suitable for bronchial asthma, asthmatic bronchitis, bronchospasm, emphysema, etc. Existing dosage forms include tablets, controlled-release tablets, aerosols, etc.

[0007] Chinese patent publication number CN116392460A discloses a salbutamol aerosol, comprising salbutamol or a pharmaceutically acceptable salt thereof, vitamin E succinate polyethylene glycol ester and a propellant.

[0008] Chinese patent publication number CN110585177A discloses a salbutamol sulfate inhalation aerosol, comprising the following components by weight: 0.1-0.3% salbutamol sulfate, 85-95% tetrafluoroethane, 0.01-0.03% oleic acid, and the balance being anhydrous ethanol. Summary of the invention

[0009] The shortcomings of the prior art are overcome, and the present invention provides a salbutamol aerosol with high stability and good efficacy through liposome encapsulation technology, and optimized formulation and preparation method.

[0010] Specifically, the salbutamol aerosol is composed of 1.0-2.0 parts by weight of salbutamol, 3.5-9.5 parts by weight of triolein, 40-120 parts by weight of ethanol, 0.5-1.0 parts by weight of meglumine, and a propellant up to 1000 parts by weight.

[0011] The preparation method of the salbutamol aerosol is:

[0012] (1) dissolving salbutamol and triolein in ethanol, removing part of the ethanol by vacuum rotary evaporation at a water bath temperature of 30°C to 40°C, adding a buffer solution to adjust the pH, continuing vacuum rotary evaporation at a water bath temperature of 40°C to 50°C until no liquid is distilled out, ultrasonicating the mixture for 10 to 30 minutes, and homogenizing under high pressure to obtain salbutamol microparticles;

[0013] (2) Add meglumine to the salbutamol microparticles, perform ultrasonic treatment for 5 to 15 minutes, fill the microparticles, and add a propellant to obtain the microparticles.

[0014] The present invention utilizes triolein to encapsulate salbutamol, effectively encapsulates the drug, protects it from the influence of the external environment, prevents drug degradation and inactivation, and can also improve the solubility of the drug, increase the solubility of the drug in the aerosol, and help the drug to achieve the treatment goal more effectively. In addition, the present invention enhances the deposition and absorption of the drug in the lungs, improves the bioavailability of the drug, increases the residence time of the drug in the lesion site, and improves the local therapeutic effect of the drug in the respiratory tract through liposome technology.

[0015] Furthermore, the buffer solution is selected from at least one of a phosphate buffer solution, a citrate buffer solution, and a carbonate buffer solution.

[0016] Furthermore, the pH of the buffer solution is adjusted to 5.5-7.0.

[0017] Furthermore, the propellant is one of dimethyl ether, tetrafluoroethane and heptafluoropropane.

[0018] Since the drug particles of the inhalation preparation need to reach the lungs to exert a therapeutic effect, the particle size needs to be controlled. The present invention uses high-pressure homogenization and ultrasound to control the particle size of the salbutamol aerosol to about 0.5 to 5 μm.

[0019] Furthermore, the salbutamol aerosol is loaded in a pressure-resistant container and a quantitative valve combination as a container. It should be noted that the pressure-resistant container and the quantitative valve are conventional supporting devices for aerosols, and suitable devices can be selected.

[0020] The invention verifies the efficacy of salbutamol aerosol in treating asthma by using animal experiments, and provides the use of the salbutamol aerosol in the preparation of a drug for treating asthma.

[0021] Compared with the prior art, the technical effects of the present invention are:

[0022] The present invention utilizes triolein to encapsulate salbutamol, thereby preventing drug degradation and inactivation and improving the local therapeutic effect of the drug in the respiratory tract.

[0023] The invention adds a proper amount of meglumine into the salbutamol aerosol prescription, significantly improves the stability of the salbutamol aerosol, effectively reduces the content of related substances, and ensures the safety of medication. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 : Changes in the total content of related substances in the accelerated tests of the salbutamol aerosols of Examples 1 to 3 and Comparative Examples 1 to 5.

[0025] Figure 2 : The change diagram of salbutamol content in the accelerated test of salbutamol aerosol of Examples 1 to 3 and Comparative Examples 1 to 5.

[0026] Figure 3 : The active ingredient content of salbutamol aerosol at different pH values.

[0027] Figure 4 : PC100 values ​​of airway responsiveness in rats of each group.

[0028] Figure 5 : The levels of immunoglobulin IgE in rats in each group.

[0029] Figure 6 : The IgG content of rats in each group. DETAILED DESCRIPTION

[0030] In order to make the purpose and technical solution of the present invention clearer, the present invention is further described below in conjunction with embodiments, but the protection scope of the present invention is not limited to these embodiments, and the embodiments are only used to explain the present invention. It should be understood by those skilled in the art that any changes or equivalent substitutions that do not deviate from the concept of the present invention are included in the protection scope of the present invention.

[0031] Example 1 Salbutamol aerosol

[0032] formula:

[0033]

[0034] Preparation method:

[0035] (1) dissolving salbutamol and triolein in ethanol, removing part of the ethanol by vacuum rotary evaporation at 35° C. in a water bath, adding a phosphate buffer solution to adjust the pH to 6.2, continuing vacuum rotary evaporation at 45° C. in a water bath until no liquid is distilled out, ultrasonicating the mixture for 20 min, and homogenizing under high pressure to obtain salbutamol microparticles;

[0036] (2) Add meglumine to the salbutamol microparticles, perform ultrasonic treatment for 10 minutes, fill the microparticles, and add a propellant to obtain the microparticles.

[0037] Example 2 Salbutamol aerosol

[0038] formula:

[0039]

[0040] Preparation method:

[0041] (1) dissolving salbutamol and triolein in ethanol, removing part of the ethanol by vacuum rotary evaporation at a water bath temperature of 30° C., adding a citrate buffer solution to adjust the pH to 5.5, continuing the vacuum rotary evaporation at a water bath temperature of 40° C. until no liquid is distilled out, ultrasonicating the mixture for 10 minutes, and homogenizing under high pressure to obtain salbutamol microparticles;

[0042] (2) Add meglumine to the salbutamol microparticles, perform ultrasonic treatment for 5 minutes, fill the microparticles, and add a propellant to obtain the microparticles.

[0043] Example 3 Salbutamol aerosol

[0044] formula:

[0045]

[0046]

[0047] Preparation method:

[0048] (1) dissolving salbutamol and triolein in ethanol, removing part of the ethanol by vacuum rotary evaporation at 40° C. in a water bath, adding a carbonate buffer solution to adjust the pH to 7.0, continuing vacuum rotary evaporation at 50° C. in a water bath until no liquid is distilled out, ultrasonicating the mixture for 30 min, and homogenizing under high pressure to obtain salbutamol microparticles;

[0049] (2) Add meglumine to the salbutamol microparticles, perform ultrasonic treatment for 15 minutes, fill the microparticles, and add a propellant to obtain the microparticles.

[0050] Comparative Example 1 Salbutamol aerosol

[0051] formula:

[0052]

[0053] Preparation method:

[0054] (1) dissolving salbutamol and triolein in ethanol, removing part of the ethanol by vacuum rotary evaporation at 35° C. in a water bath, adding a phosphate buffer solution to adjust the pH to 6.2, continuing vacuum rotary evaporation at 45° C. in a water bath until no liquid is distilled out, ultrasonicating the mixture for 20 min, and homogenizing under high pressure to obtain salbutamol microparticles;

[0055] (2) The salbutamol microparticles are sonicated for 10 minutes, filled, and added with a propellant to obtain the salbutamol microparticles.

[0056] Comparative Example 2 Salbutamol aerosol

[0057] formula:

[0058]

[0059] Preparation method:

[0060] (1) dissolving salbutamol and lecithin in ethanol, removing part of the ethanol by vacuum rotary evaporation at 35° C. in a water bath, adding a phosphate buffer solution to adjust the pH to 6.2, continuing vacuum rotary evaporation at 45° C. in a water bath until no liquid is distilled out, ultrasonicating the mixture for 20 min, and homogenizing under high pressure to obtain salbutamol microparticles;

[0061] (2) Add meglumine to the salbutamol microparticles, perform ultrasonic treatment for 10 minutes, fill the microparticles, and add a propellant to obtain the microparticles.

[0062] Comparative Example 3 Salbutamol aerosol

[0063] formula:

[0064]

[0065] Preparation method:

[0066] (1) dissolving salbutamol, triolein, and meglumine in ethanol, removing part of the ethanol by vacuum rotary evaporation at a water bath temperature of 35° C., adding phosphate buffer solution to adjust the pH to 6.2, continuing vacuum rotary evaporation at a water bath temperature of 45° C. until no liquid is distilled out, ultrasonicating the mixture for 20 minutes, and homogenizing under high pressure to obtain salbutamol microparticles;

[0067] (2) The salbutamol microparticles are sonicated for 10 minutes, filled, and added with a propellant to obtain the salbutamol microparticles.

[0068] Comparative Example 4 Salbutamol aerosol

[0069] formula:

[0070]

[0071] Preparation method:

[0072] (1) dissolving salbutamol in ethanol and ultrasonicating for 20 min to obtain salbutamol microparticles;

[0073] (2) Add meglumine to the salbutamol microparticles, perform ultrasonic treatment for 10 minutes, fill the microparticles, and add a propellant to obtain the microparticles.

[0074] Comparative Example 5 Commercially available salbutamol aerosol (National Medicine Standard H23020333)

[0075] Determination of related substances in salbutamol aerosol

[0076] The salbutamol aerosols of Examples 1 to 3 and the salbutamol aerosols of Comparative Examples 1 to 5 were subjected to an accelerated test at a temperature of 40°C ± 2°C, a relative humidity of 75% ± 5%, and an illumination of 4500 lx ± 500 lx. Samples were taken at the first day, the end of the second month, the end of the third month, and the end of the sixth month, respectively, to determine the contents of salbutamol-related substances.

[0077] The related substances of salbutamol aerosol are determined according to the HPLC method (General Rule 0512) of the 2020 edition of the Chinese Pharmacopoeia. Test solution: Take an appropriate amount of this product, add the mobile phase to dissolve and dilute to make a solution containing approximately 2 mg per 1 ml (determined within 12 hours). Control solution: Accurately measure 1 ml of the test solution, place it in a 100 ml volumetric flask, dilute to the scale with the mobile phase, and shake well (determined within 12 hours). System suitability solution: Take an appropriate amount of terbutaline sulfate and salbutamol, add the mobile phase to dissolve and dilute to make a solution containing approximately 0.2 mg per 1 ml. Chromatographic conditions: Octylsilane bonded silica gel is used as filler; sodium heptane sulfonate solution [take 2.87g of sodium heptane sulfonate and 2.5g of potassium dihydrogen phosphate, dissolve in water and dilute to 1000ml, adjust the pH value to 3.65 with phosphoric acid solution (1→2)]-acetonitrile (78:22) is used as mobile phase; detection wavelength is 220nm; injection volume is 20μl). System suitability requirements: In the system suitability solution chromatogram, the separation between the salbutamol peak and the terbutaline peak should meet the requirements. Determination method: Accurately measure the test solution and the control solution, inject them into the liquid chromatograph respectively, and record the chromatogram until 25 times the retention time of the main component peak. Limit: If there are impurity peaks in the chromatogram of the test solution, the sum of the areas of each impurity peak shall not be greater than the main peak area of ​​the control solution (1.0%).

[0078] Figure 1 The graph is a graph showing the changes in the total content of related substances in the accelerated test of the salbutamol aerosols of Examples 1 to 3 and Comparative Examples 1 to 5. The impurity content of the salbutamol aerosols of Examples 1 to 3 is significantly lower than that of the salbutamol aerosols of the Comparative Examples, which can reduce the toxicity or other adverse effects that may occur during the use of the drug. In particular, asthma cannot be completely cured at present, and the condition needs to be controlled for a long time by drugs, and the safety of salbutamol aerosol is extremely important.

[0079] Determination of the active ingredient content in salbutamol aerosol

[0080] The salbutamol aerosols of Examples 1 to 3 and the salbutamol aerosols of Comparative Examples 1 to 5 were placed under conditions of 40°C ± 2°C, 75% ± 5% relative humidity, and 4500 lx ± 500 lx for accelerated testing, and samples were taken on the first day, at the end of the second month, at the end of the third month, and at the end of the sixth month to determine the salbutamol content. The salbutamol content was determined according to the high performance liquid chromatography method (General Rule 0512) of the 2020 edition of the Chinese Pharmacopoeia.

[0081] Figure 2The diagram is a change in the salbutamol content in the accelerated test of the salbutamol aerosols of Examples 1 to 3 and Comparative Examples 1 to 5. Compared with the salbutamol aerosols of Comparative Examples 1 to 5, the salbutamol aerosols of Examples 1 to 3 show higher stability of the active ingredient content, and the high-purity active ingredient is more likely to show the expected therapeutic effect, because there are no impurities that affect the interaction between the drug and the organism, ensuring that the drug can accurately exert its effect.

[0082] Single factor experiment to explore the effect of pH on the stability of the active ingredient content of salbutamol aerosol

[0083] The single factor variable was pH value, which was 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, and 8.0 respectively. The other experimental conditions were the same as the salbutamol aerosol in Example 1. Each group of salbutamol aerosol was prepared and tested under the same conditions under the determination of the content of the active ingredient in the salbutamol aerosol to detect the salbutamol content at the end of the 6th month.

[0084] Figure 3 The results show that the pH value in the liposome preparation method has a great influence on the stability of the active ingredient content of salbutamol aerosol. Under the reasonable coordination and regulation of other factors, the pH value of 5.5-7.0 has a better effect on the stabilization of salbutamol.

[0085] Pharmacodynamics of salbutamol aerosol in the treatment of asthmatic rats

[0086] 1. Experimental Animals

[0087] SD rats SPF grade, 180-220 g, experimental animal license number: SYXK(Lu)20180008, were adaptively fed under standard conditions for 1 week before the experiment.

[0088] 2. Modeling method and grouping

[0089] The asthma model of rats was established by sensitization and stimulation with ovalbumin (OVA). Several rats were sensitized by intraperitoneal injection of 1 ml of OVA sensitization solution on the first and eighth days of modeling. Ten rats in the blank group were intraperitoneally injected with physiological saline instead of OVA sensitization solution at the same time and the same dose. From the 15th day, except for the blank group, the rats were placed in the nebulizer and stimulated by nebulization with 1% OVA nebulizer solution for 20 minutes per day for a total of 7 days. The stimulation was successful when the rats showed accelerated breathing, cyanosis of the lips, abdominal muscle spasms, nodding breathing and unstable standing. The blank group rats were nebulized with physiological saline instead of OVA nebulizer solution, and the time and method were the same as those of the asthma model group rats. The rats with successful modeling were divided into the model group, the embodiment 1 group, the embodiment 2 group, the embodiment 3 group, the comparative example 4 group, and the comparative example 5 group, with 10 rats in each group.

[0090] 3. Drug administration

[0091] The rats in each treatment group were given the corresponding drugs by nebulization inhalation, and the rats in the model group and the blank group were given normal saline by nebulization inhalation. The drug was administered twice a day for 7 days, with a dose of 9 μg / kg.

[0092] 4. Statistical analysis

[0093] SPSS 22.0 software was used to analyze the data. The comparison among multiple groups was performed by one-way analysis of variance, and the comparison between two groups was performed by independent sample T test. P < 0.05 was considered statistically significant.

[0094] 5. Observation indicators and experimental results

[0095] 5.1 Airway responsiveness measurement

[0096] The airway responsiveness of rats in each group was measured using a noninvasive whole body plethysmography system. After the rats were sealed in a plethysmography box for 5 minutes, bronchial stimulation was performed using methacholine (Mch) solution to observe the changes in the airway resistance parameter values ​​under Mch stimulation. The Mch stimulation concentration when the airway responsiveness increased by 2 times the baseline value, i.e. PC100, was used as the evaluation index.

[0097] Figure 4 is the PC100 value of airway reactivity of each group of rats. Compared with the rats in comparative example 5, the PC100 of the rats in Example 1, Example 2, and Example 3 were all increased, with significant differences, P < 0.01. This shows that the salbutamol aerosols of Examples 1 to 3 of the present invention are more effective in treating asthma than the commercially available salbutamol aerosols. The PC100 value of airway reactivity of the salbutamol aerosol in comparative example 4 is slightly lower than that of the PC100 value of airway reactivity in comparative example 5, indicating that when salbutamol is made into liposomes and then prepared into aerosols for administration, the drug is easier to be absorbed by epithelial cells with lipid membrane properties, and the effect is better and ideal.

[0098] 5.2 Determination of immunoglobulins

[0099] Anesthetize, collect blood through the abdominal aorta, centrifuge at 3000r / min for 10min, draw the supernatant, divide it into portions, store in a -20℃ refrigerator, and wait for testing. Add ovalbumin to each well of a 96-well cell culture plate at 4℃ overnight, rinse with PBS buffer, then add 1% BSA, 150μL per well, culture at 37℃ for 1h, and rinse with PBS buffer. Dilute the serum that has been taken with buffer, add 200μL to each well, culture at 37℃ for 3h, and rinse with PBS buffer. Dilute IgE and IgG antibodies with buffer, add 100μL to each well, culture at 37℃ for 2h, then rinse with PBS buffer, finally take the substrate and make it into 1g / L with carbonate buffer, add 100μL to each well, culture for 15min (IgG) and 12h (IgE), respectively, and measure the absorbance (A) at wavelengths of 405nm and 492nm.

[0100] Figure 5 is the immunoglobulin IgE content of rats in each group, Figure 6 Compared with the rats in the comparative example 5, the immunoglobulins IgE and IgG of the rats in the example 1 group, the example 2 group and the example 3 group were significantly different, P < 0.01.

Claims

1. A salbutamol aerosol, characterized in that: The salbutamol aerosol is composed of the following ingredients: 1.0-2.0 parts by weight of salbutamol, 3.5-9.5 parts by weight of triolein, 40-120 parts by weight of ethanol, an appropriate amount of buffer solution, 0.5-1.0 parts by weight of meglumine, and a propellant added to make up to 1000 parts by weight; The preparation method of the salbutamol aerosol is: (1) dissolving salbutamol and triolein in ethanol, removing part of the ethanol by a first vacuum rotary evaporation, adding a buffer solution to adjust the pH to 5.5-7.0, performing a second vacuum rotary evaporation until no liquid is distilled out, ultrasonicating the mixture for 10-30 minutes, and homogenizing under high pressure to obtain salbutamol microparticles; (2) Add meglumine to the salbutamol microparticles, perform ultrasonic treatment for 5 to 15 minutes, fill the microparticles, and add a propellant to obtain the microparticles.

2. The salbutamol aerosol according to claim 1, characterized in that The propellant is one of dimethyl ether, tetrafluoroethane and heptafluoropropane.

3. The salbutamol aerosol according to claim 1, characterized in that The water bath temperature of the first vacuum rotary evaporation is 30°C to 40°C.

4. The salbutamol aerosol according to claim 1, characterized in that The water bath temperature of the second vacuum rotary evaporation is 40°C to 50°C.

5. The salbutamol aerosol according to claim 1, characterized in that The buffer solution is selected from at least one of a phosphate buffer solution, a citrate buffer solution, and a carbonate buffer solution.

6. The salbutamol aerosol according to claim 1, characterized in that The particle size of the salbutamol aerosol is 0.5 to 5 μm.

7. The salbutamol aerosol according to claim 1, characterized in that The salbutamol aerosol is loaded in a pressure-resistant container and a quantitative valve combination as a container.

8. Use of the salbutamol aerosol according to claim 1 in preparing a drug for treating asthma.

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