Sodium pyruvate aerosol inhalation preparation and preparation method thereof

Abstract

The invention belongs to the field of biological medicine, and particularly relates to a sodium pyruvate aerosol inhalation preparation and a preparation method thereof. The aerosol inhalation preparation is prepared from the following raw materials in parts by weight: 0.04 to 0.09 part of baicalin-berbamine / sodium pyruvate clathrate compound freeze-dried powder and 100 parts of normal saline, the preparation process of the baicalin-berbamine comprises the following steps: reacting berbamine with 4-chlorphenyl glycidyl ether to obtain an intermediate A, mixing the intermediate A with a baicalin solution, reacting in the presence of a catalyst, and purifying after the reaction is completed to obtain the baicalin-berbamine. The sodium pyruvate aerosol inhalation preparation can effectively relieve asthma symptoms, and has excellent antibacterial activity, stability and biological activity.

Description

Technical Field

[0001] The invention belongs to the field of biomedicine, and specifically relates to a sodium pyruvate atomized inhalation preparation and a preparation method thereof. Background Art

[0002] Bronchial asthma, referred to as asthma, is a common chronic inflammatory respiratory disease. The pathogenesis includes: chronic airway inflammation, airway hyperresponsiveness, airway neural dysregulation, genetic mechanism, respiratory viral infection, etc. The main characteristics are repeated episodes of wheezing, shortness of breath, chest tightness or cough in the airways. In severe cases, dyspnea and hypoxemia may occur. It often occurs or worsens at night and early morning. The onset of asthma is related to genetic and environmental factors. Among them, 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 asthma include exercise-induced, drug-induced, occupational and allergic. The symptoms of asthma are mainly wheezing, shortness of breath, chest tightness or cough.

[0003] Sodium pyruvate is the sodium salt of pyruvic acid. Pyruvic acid is a three-carbon keto acid, a colorless liquid with an acetic acid-like odor. It is an important intermediate metabolite in the process of sugar metabolism and a precursor of a variety of useful compounds. Therefore, it has a wide range of uses in industries such as chemical, pharmaceutical and agricultural chemicals and scientific research. With the deepening of research, scholars have found that sodium pyruvate can also be used as a drug for the treatment of asthma. Existing sodium pyruvate preparations have the problem of unsatisfactory effect on improving asthma symptoms, and thus cannot effectively and quickly relieve asthma symptoms.

[0004] Traditional Chinese medicine is favored by people due to its advantages such as high safety and low toxicity and side effects. Baicalin is a flavonoid compound extracted and separated from the dried root of Scutellaria baicalensis Georgi, a dicotyledonous plant of the Labiatae family. It has significant biological activity, antibacterial, anti-inflammatory, purging fire and detoxifying, hemostatic, tocolytic, anti-allergic and antispasmodic effects. It is also a specific inhibitor of mammalian liver salivary enzymes, has the function of regulating certain diseases, and also has strong anti-cancer physiological efficacy.

[0005] Berbamine is a dibenzylisoquinoline alkaloid extracted from Chinese medicinal plants such as Coptis chinensis, Berberis chinensis, and Berberis tenuifolia. It has pharmacological effects such as anti-arrhythmia, anti-tumor, and promotion of leukocyte proliferation. At present, there is no evidence that the above two components derived from Chinese medicine are used to relieve asthma symptoms after chemical grafting modification. Summary of the invention

[0006] The first object of the present invention is to provide a sodium pyruvate aerosol inhalation preparation, which has high stability and strong antibacterial performance.

[0007] The second object of the present invention is to provide a method for preparing a sodium pyruvate aerosol inhalation preparation, which is simple and easy to practice.

[0008] In order to achieve the above object, the technical solution adopted by the present invention is: A sodium pyruvate aerosol inhalation preparation, comprising the following raw materials in parts by weight: 0.04-0.09 parts of baicalin-berbamine / sodium pyruvate inclusion compound freeze-dried powder, 100 parts of normal saline; The preparation process of baicalin-berberamine is as follows: Berbamine reacts with 4-chlorophenyl glycidyl ether to obtain intermediate A, and the intermediate A is mixed with baicalin solution and reacted under catalyst conditions. After the reaction is completed, baicalin-berbamine is purified to obtain baicalin-berbamine.

[0009] Furthermore, the molar ratio of the intermediate A, baicalin and catalyst is 1:0.25-0.5:15-20, and the catalyst is triethylamine.

[0010] Furthermore, the conditions for the reaction of the intermediate A with the baicalin solution are: reacting at 75-90° C. for 30-60 min.

[0011] Furthermore, the molar ratio of berbamine to 4-chlorophenyl glycidyl ether is 1:1.2-1.5, and the reaction conditions are: reacting at 55-70° C. for 2-4 hours.

[0012] Furthermore, the preparation method of the baicalin-berbamine / sodium pyruvate inclusion compound freeze-dried powder includes: using a polymer as a carrier, and adopting an emulsified solvent volatilization method to load sodium pyruvate and baicalin-berbamine onto the polymer to obtain nanoparticles; performing surface hydrophilic modification on the nanoparticles, and adding the modified nanoparticles to a lyophilization protective agent solution, and freeze-drying to obtain the nanoparticles.

[0013] Furthermore, the mass ratio of the sodium pyruvate, baicalin-berberamine and polymer is 1:1-10:1-10, and the polymer is selected from one of polylactic acid, polylactic acid-glycolic acid copolymer and polycaprolactone.

[0014] Furthermore, the concentration of the hydrophilic compound used in the hydrophilic modification is 0.3-0.6 mol / L, the hydrophilic compound is selected from polyethylene glycol or polyvinyl pyrrolidone, and the lyophilization protective agent is composed of mannitol and trehalose in a mass ratio of 1:1-2.

[0015] Furthermore, the freeze-drying is specifically as follows: first pre-freeze at normal pressure -30°C~-40°C for 2~4h; then evacuate and keep warm at 5~30Pa, -1~-10°C for 25~30h; then keep warm at 0~5°C for 1~2h; then heat to 25~30°C and keep warm for 12-16h.

[0016] Furthermore, the specific operation of the emulsified solvent volatilization method includes: adding the oil phase to the water phase under stirring, emulsifying and solidifying, centrifuging and washing; the water phase is a polyvinyl alcohol solution, and the oil phase is selected from one of a polylactic acid solution, a polylactic acid-glycolic acid copolymer solution, and a polycaprolactone solution; Wherein, the mass concentration of the polyvinyl alcohol in the water phase is 0.8-1.5%.

[0017] A method for preparing a sodium pyruvate aerosol inhalation preparation comprises the steps of compounding baicalin-berbamine / sodium pyruvate inclusion compound freeze-dried powder with physiological saline according to the above ratio to obtain the preparation.

[0018] The beneficial technical effects of the present invention are: 1. The present invention provides a sodium pyruvate aerosol inhalation preparation, the main component of which is baicalin-berbamine / sodium pyruvate inclusion compound freeze-dried powder. Experiments have found that the direct combination of baicalin, berbamine and sodium pyruvate is better than the use of sodium pyruvate alone, indicating that the above two ingredients play a positive role in relieving asthma symptoms. The inventor further modified the structure of berbamine by reacting with 4-chlorophenyl glycidyl ether and then grafted it with baicalin, and used the above compound in combination with sodium pyruvate, and the effect was better than the combination of baicalin, berbamine and sodium pyruvate.

[0019] 2. The present invention adopts the emulsified solvent volatilization method to load baicalin-berbamine and sodium pyruvate onto a polymer carrier to enhance the absorption performance and stability of the drug in the lungs.

[0020] 3. In addition, the sodium pyruvate aerosol inhalation preparation of the present invention can effectively relieve asthma symptoms and has excellent antibacterial properties, stability and biological activity. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 is the baicalin-berberamine in Example 1 of the present invention 1 HNMR spectrum. DETAILED DESCRIPTION

[0022] The following is a further detailed description of the present invention in conjunction with specific preferred embodiments, and it cannot be determined that the specific implementation of the present invention is limited to these descriptions. For those of ordinary skill in the technical field to which the present invention belongs, without departing from the concept of the present invention, several simple deductions or replacements can also be made, all of which should be deemed to belong to the protection scope of the present invention. The specific conditions not indicated in the embodiments are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used, unless otherwise specified, are conventional products obtained through commercial channels.

[0023] Example Example 1 The present embodiment provides a sodium pyruvate aerosol inhalation preparation, which is composed of 0.06 parts of baicalin-berbamine / sodium pyruvate inclusion compound freeze-dried powder and 100 parts of normal saline.

[0024] Wherein, the preparation process of baicalin-berberamine is as follows: According to the molar ratio of berbamine to 4-chlorophenyl glycidyl ether being 1:1.3, berbamine and 4-chlorophenyl glycidyl ether are reacted at 60°C for 3 hours. After the reaction is completed, the reaction solution is cooled to room temperature, and unreacted 4-chlorophenyl glycidyl ether is removed by reduced pressure distillation to obtain intermediate A; According to the molar ratio of intermediate A, baicalin and triethylamine of 1:0.4:17, intermediate A was mixed with baicalin solution (solvent: DMF), and reacted at 80°C for 45 minutes in the presence of triethylamine. After the reaction was completed, the reaction solution was immediately filtered, cooled to room temperature, and the pH was adjusted to 2 with hydrochloric acid. After crystallization, it was filtered to obtain the product.

[0025] The NMR characterization results of the obtained baicalin-berberamine are ( Figure 1 shown): 1H-NMR (400MHz, DMSO) δ = 2.27 (s, 12H), 2.65 (m, 4H), 2.75-2.83 (m, 16H), 3.07 (m, 4H), 3.72-3.73 (d, 12H), 3. 84 (s, 6H), 3.96 (m, 4H), 4.13 (m, 1H), 4.21-4.30 (m, 10H), 4.52 (s, 1H), 4.61 (m, 2H), 4.78 (s, 1H), 4.98 (d, 1H) ,5.78-5.81 (m, 3H),6.17 (s, 1H),6.61 (s, 2H),6.72 (s, 1H),6.81-6.86 (m, 6H),6.94-6.99 (s, 8H),7.02-7.10 (d, 6H),7.32-7.34 (d, 6H),7.47-7.49 (d, 3H),7.78 (m, 2H),8.74 (s, 1H),12.87 (s, 1H). The above results confirmed that the obtained product was the target product.

[0026] The preparation method of the above-mentioned baicalin-berbamine / sodium pyruvate inclusion compound freeze-dried powder is as follows: Water phase: dissolve polyvinyl alcohol in deionized water to obtain a water phase, wherein the mass concentration of polyvinyl alcohol in the water phase is 1%, and set aside; Oil phase: sodium pyruvate, baicalin-berberamine, and polylactic acid are added to a polylactic acid solution (solvent is dichloromethane) at a mass ratio of 1:6:6, and a stirring device is turned on for stirring to obtain an oil phase. The concentration of polylactic acid in the oil phase is 0.1 g / mL; Under mechanical stirring, the oil phase is added to the aqueous phase for emulsification and solidification. After the solidification is completed, the reaction solution is centrifuged and washed to obtain nanoparticles; the nanoparticles are dispersed in a 0.4 mol / L polyethylene glycol aqueous solution Z, and then added to a lyophilization protective agent solution, which is composed of a mannitol and trehalose aqueous solution in a mass ratio of 1:1, and then pre-frozen at normal pressure of -35°C for 3 hours; then evacuated, kept warm at 15Pa and -6°C for 27 hours; then kept warm at 2°C for 1.5 hours; then heated to 27°C and kept warm for 14 hours to obtain the product.

[0027] A method for preparing a sodium pyruvate aerosol inhalation preparation is as follows: The baicalin-berbamine / sodium pyruvate inclusion compound freeze-dried powder is compounded with normal saline according to the above ratio to obtain the product.

[0028] Example 2 This embodiment provides a sodium pyruvate aerosol inhalation preparation, which consists of 0.09 parts of baicalin-berbamine / sodium pyruvate inclusion compound freeze-dried powder and 100 parts of normal saline.

[0029] Wherein, the preparation process of baicalin-berberamine is as follows: According to the molar ratio of berbamine to 4-chlorophenyl glycidyl ether being 1:1.5, berbamine and 4-chlorophenyl glycidyl ether are reacted at 55°C for 2h. After the reaction is completed, the reaction solution is cooled to room temperature, and unreacted 4-chlorophenyl glycidyl ether is removed by reduced pressure distillation to obtain intermediate A; According to the molar ratio of intermediate A, baicalin and triethylamine of 1:0.5:15, intermediate A was mixed with baicalin solution (solvent: DMF), and reacted at 75°C for 60 min in the presence of triethylamine. After the reaction was completed, the reaction solution was immediately filtered and cooled to room temperature. The pH was adjusted to 2 with hydrochloric acid, and the product was obtained after crystallization and filtration.

[0030] The preparation method of the above-mentioned baicalin-berbamine / sodium pyruvate inclusion compound freeze-dried powder is as follows: Water phase: dissolve polyvinyl alcohol in deionized water to obtain a water phase, wherein the mass concentration of polyvinyl alcohol in the water phase is 0.8%, and set aside; Oil phase: sodium pyruvate, baicalin-berberamine and polylactic acid-co-glycolic acid were added to the polylactic acid-co-glycolic acid solution (solvent: dichloromethane) at a mass ratio of 1:1:1, and the stirring device was turned on for stirring to obtain an oil phase. The concentration of polylactic acid in the oil phase was 0.1 g / mL; Under mechanical stirring, the oil phase is added to the aqueous phase for emulsification and solidification. After the solidification, the reaction solution is centrifuged and washed to obtain nanoparticles; the nanoparticles are dispersed in a 0.6 mol / L polyethylene glycol aqueous solution, and then added to a lyophilization protective agent solution, which is composed of a mannitol and trehalose aqueous solution in a mass ratio of 1:1.5, and then pre-frozen at normal pressure of -30°C for 4 hours; then evacuated and kept warm at 5Pa and -10°C for 25 hours; then kept warm at 5°C for 1 hour; then heated to 30°C and kept warm for 12 hours to obtain the product.

[0031] A method for preparing a sodium pyruvate aerosol inhalation preparation is as follows: The baicalin-berbamine / sodium pyruvate inclusion compound freeze-dried powder is compounded with normal saline according to the above ratio to obtain the product.

[0032] Example 3 The present embodiment provides a sodium pyruvate aerosol inhalation preparation, which is composed of 0.04 parts of baicalin-berbamine / sodium pyruvate inclusion compound freeze-dried powder and 100 parts of normal saline.

[0033] Wherein, the preparation process of baicalin-berberamine is as follows: According to the molar ratio of berbamine to 4-chlorophenyl glycidyl ether being 1:1.2, berbamine and 4-chlorophenyl glycidyl ether are reacted at 70°C for 4 hours. After the reaction is completed, the reaction solution is cooled to room temperature, and unreacted 4-chlorophenyl glycidyl ether is removed by reduced pressure distillation to obtain intermediate A; According to the molar ratio of intermediate A, baicalin and triethylamine of 1:0.25:20, intermediate A was mixed with baicalin solution (solvent: DMF), and reacted at 90°C for 30 min in the presence of triethylamine. After the reaction was completed, the reaction solution was immediately filtered and cooled to room temperature. The pH was adjusted to 2 with hydrochloric acid, and the product was obtained after crystallization and filtration.

[0034] The preparation method of the above-mentioned baicalin-berberamine-sodium pyruvate inclusion compound freeze-dried powder is as follows: Water phase: dissolve polyvinyl alcohol in deionized water to obtain a water phase, wherein the mass concentration of polyvinyl alcohol in the water phase is 1.5%, and set aside; Oil phase: sodium pyruvate, baicalin-berberamine, and polycaprolactone are added to the polycaprolactone solution (solvent is dichloromethane) at a mass ratio of 1:10:10, and a stirring device is turned on for stirring to obtain an oil phase. The concentration of polylactic acid in the oil phase is 0.1 g / mL; Under mechanical stirring, the oil phase is added to the aqueous phase for emulsification and solidification. After the solidification, the reaction solution is centrifuged and washed to obtain nanoparticles; the nanoparticles are dispersed in a 0.3 mol / L polyvinyl pyrrolidone aqueous solution, and then added to a freeze-drying protective agent solution, which is composed of a mannitol and trehalose aqueous solution in a mass ratio of 1:2, and then pre-frozen at normal pressure of -40°C for 2h; then evacuated and kept warm at 30Pa and -1°C for 30h; then kept warm at 0°C for 2h; then heated to 25°C and kept warm for 16h to obtain the product.

[0035] A method for preparing a sodium pyruvate aerosol inhalation preparation is as follows: The baicalin-berbamine / sodium pyruvate inclusion compound freeze-dried powder is compounded with normal saline according to the above ratio to obtain the product.

[0036] Comparative Example Comparative Example 1 Comparative Example 1 provides a sodium pyruvate aerosol inhalation preparation, which consists of 0.06 parts of baicalin / sodium pyruvate inclusion compound freeze-dried powder and 100 parts of normal saline.

[0037] The preparation method of the above-mentioned baicalin / sodium pyruvate inclusion compound freeze-dried powder is as follows: Water phase: dissolve polyvinyl alcohol in deionized water to obtain a water phase, wherein the mass concentration of polyvinyl alcohol in the water phase is 1%, and set aside; Oil phase: according to the mass ratio of sodium pyruvate, baicalin and polylactic acid of 1:6:6, sodium pyruvate and baicalin are added to the polylactic acid solution (the solvent is dichloromethane), and the stirring device is turned on for stirring to obtain an oil phase. The concentration of polylactic acid in the oil phase is 0.1 g / mL, and it is set aside; Under mechanical stirring, the oil phase is added to the water phase for emulsification and solidification. After solidification, the reaction solution is centrifuged and washed to obtain nanoparticles; the nanoparticles are dispersed in a 0.4 mol / L polyethylene glycol aqueous solution, and then added to a freeze-drying protective agent solution, which is composed of a mannitol and trehalose aqueous solution with a mass ratio of 1:1, and then pre-frozen at normal pressure of -35°C for 3 hours; then vacuumed, kept warm at 15Pa and -6°C for 27 hours; then kept warm at 2°C for 1.5 hours; then heated to 27°C and kept warm for 14 hours to obtain the product. The rest is the same as in Example 1.

[0038] Comparative Example 2 Comparative Example 2 provides a sodium pyruvate aerosol inhalation preparation, which consists of 0.06 parts of berbamine / sodium pyruvate inclusion compound lyophilized powder and 100 parts of normal saline.

[0039] The preparation method of the berbamine / sodium pyruvate inclusion compound freeze-dried powder is as follows: Water phase: dissolve polyvinyl alcohol in deionized water to obtain a water phase, wherein the mass concentration of polyvinyl alcohol in the water phase is 1%, and set aside; Oil phase: according to the mass ratio of sodium pyruvate, berbamine and polylactic acid of 1:6:6, sodium pyruvate and berbamine are added to the polylactic acid solution (the solvent is dichloromethane), and the stirring device is turned on for stirring to obtain an oil phase. The concentration of polylactic acid in the oil phase is 0.1 g / mL, and it is set aside; Under mechanical stirring, the oil phase is added to the water phase for emulsification and solidification. After solidification, the reaction solution is centrifuged and washed to obtain nanoparticles; the nanoparticles are dispersed in a 0.4 mol / L polyethylene glycol aqueous solution, and then added to a freeze-drying protective agent solution, which is composed of a mannitol and trehalose aqueous solution with a mass ratio of 1:1, and then pre-frozen at normal pressure of -35°C for 3 hours; then vacuumed, kept warm at 15Pa and -6°C for 27 hours; then kept warm at 2°C for 1.5 hours; then heated to 27°C and kept warm for 14 hours to obtain the product. The rest is the same as in Example 1.

[0040] Comparative Example 3 Comparative Example 3 provides a sodium pyruvate aerosol inhalation preparation, which consists of 0.06 parts of baicalin / berbamine / sodium pyruvate inclusion compound freeze-dried powder and 100 parts of normal saline.

[0041] The preparation method of the above-mentioned baicalin / berbamine / sodium pyruvate inclusion compound freeze-dried powder is as follows: Water phase: dissolve polyvinyl alcohol in deionized water to obtain a water phase, wherein the mass concentration of polyvinyl alcohol in the water phase is 1%, and set aside; Oil phase: according to the mass ratio of sodium pyruvate, baicalin and berbamine mixture to polylactic acid of 1:6:6, add sodium pyruvate, baicalin and berbamine mixture to polylactic acid solution (solvent is dichloromethane), and start stirring equipment to stir to obtain oil phase. The concentration of polylactic acid in the oil phase is 0.1 g / mL, and it is set aside; Under mechanical stirring, the oil phase is added to the water phase for emulsification and solidification. After solidification, the reaction solution is centrifuged and washed to obtain nanoparticles; the nanoparticles are dispersed in a 0.4 mol / L polyethylene glycol aqueous solution, and then added to a freeze-drying protective agent solution, which is composed of a mannitol and trehalose aqueous solution with a mass ratio of 1:1, and then pre-frozen at normal pressure -35°C for 3 hours; then vacuumed, kept warm at 15Pa and -6°C for 27 hours; then kept warm at 2°C for 1.5 hours; then heated to 27°C and kept warm for 14 hours to obtain the obtained nanoparticles. The rest is the same as in Example 1.

[0042] Comparative Example 4 Comparative Example 4 provides a sodium pyruvate aerosol inhalation preparation, which consists of 0.06 parts of sodium pyruvate inclusion compound lyophilized powder and 100 parts of normal saline.

[0043] The preparation method of the above-mentioned sodium pyruvate inclusion compound lyophilized powder is as follows: Water phase: dissolve polyvinyl alcohol in deionized water to obtain a water phase, wherein the mass concentration of polyvinyl alcohol in the water phase is 1%, and set aside; Oil phase: add sodium pyruvate to polylactic acid solution (solvent is dichloromethane) at a mass ratio of 1:6. The concentration of polylactic acid in the oil phase is 0.1 g / mL. Set aside. Under mechanical stirring, the oil phase is added to the water phase for emulsification and solidification. After solidification, the reaction solution is centrifuged and washed to obtain nanoparticles; the nanoparticles are dispersed in a 0.4 mol / L polyethylene glycol aqueous solution, and then added to a freeze-drying protective agent solution, which is composed of a mannitol and trehalose aqueous solution with a mass ratio of 1:1, and then pre-frozen at normal pressure -35°C for 3 hours; then vacuumed, kept warm at 15Pa and -6°C for 27 hours; then kept warm at 2°C for 1.5 hours; then heated to 27°C and kept warm for 14 hours to obtain the obtained nanoparticles. The rest is the same as in Example 1.

[0044] Experimental Example 1 The safety of the sodium pyruvate aerosol inhalation preparation of Example 1 was tested, and the process was as follows: Experimental subjects 20 guinea pigs, weighing 290-320 g; Experimental Grouping Negative control group: 0.9% sodium chloride injection by mass volume percentage; Experimental process: A nebulizer was used for aerosol inhalation administration (sodium pyruvate dosage was 8 mg / kg). Each group was administered for 14 consecutive days, once a day, with a dosing volume of 1 mL / kg. Observation was performed during the dosing period and the recovery period. All animals were killed 24 hours after the last administration. After dissection, the irritation reactions of the animal's lips, palate, tongue, nose, pharynx, larynx, trachea, and lungs were observed with the naked eye to observe whether there were irritation reactions such as congestion and redness. The final evaluation was performed based on naked eye observation. The results are shown in Table 1.

[0045] Table 1 As shown in Table 1, under the experimental conditions, after the guinea pigs were given the sodium pyruvate aerosol inhalation preparation by aerosol inhalation, no obvious irritation changes were observed in the respiratory system of the guinea pigs, which verified the safety of the sodium pyruvate aerosol inhalation preparation of the present invention.

[0046] Experimental Example 2 Experimental Example 2 The encapsulation efficiency of the sodium pyruvate products of Example 1 and Comparative Examples 1-4 was tested, and the testing process was as follows: Select 255nm as the detection wavelength of sodium pyruvate, determine its content by UV method, and establish the standard curve of sodium pyruvate; In the process of preparing nanoparticles, 1 mL of the reaction solution after solid-liquid separation in Example 1 and Comparative Examples 1-4 was taken, the volume of the supernatant was measured after centrifugation, and the absorbance was substituted into the standard curve to obtain the content of free sodium pyruvate in the supernatant, and the encapsulation rate of sodium pyruvate was calculated. The calculation formula is: encapsulation rate = (initial addition amount of sodium pyruvate - mass of free sodium pyruvate in supernatant) / initial addition amount of sodium pyruvate × 100%. For specific results, see Table 2.

[0047] Table 2 It can be seen from Table 2 that the sodium pyruvate inclusion compound prepared in the present invention has a higher encapsulation rate.

[0048] Experimental Example 3 The antibacterial effects of the aerosol inhalation preparations of Example 1 and Comparative Examples 1-4 were tested as follows: Staphylococcus aureus solution (10 8CFU / mL, 100 μL) and 900 μL of the aerosol inhalation preparations of Example 1 and Comparative Examples 1-4 were added to different shaking tubes and placed in a constant temperature shaker at 37°C. After 12 hours, the bacterial solution was diluted to 10 with PBS (pH 7.4). 5 CFU / mL, take 80μL and drop it onto the agar plate and spread it evenly, then observe and record the number of colonies after culturing for 24h, set up a blank control group (PBS), and calculate the inhibition rate according to the following formula: (number of colonies in the blank group - number of colonies in the experimental group) / number of colonies in the blank group × 100%. The results are shown in Table 3.

[0049] The beta-hemolytic streptococcus solution (10 8 CFU / mL, 100 μL) and 900 μL of the aerosol inhalation preparations of Example 1 and Comparative Examples 1-4 were added to different shaking tubes and placed in a constant temperature shaker at 37°C. After 12 hours, the bacterial solution was diluted to 10 with PBS (pH 7.4). 5 CFU / mL, take 80μL and drop it onto the agar plate and spread it evenly, then observe and record the number of colonies after culturing for 24h, set up a blank control group (PBS), and calculate the inhibition rate according to the following formula: (number of colonies in the blank group - number of colonies in the experimental group) / number of colonies in the blank group × 100%. The results are shown in Table 3.

[0050] Table 3 As shown in Table 3, the antibacterial effect of the aerosol preparation of Example 1 is significantly higher than that of Comparative Examples 1-4. Analysis shows that the lyophilized powder in the aerosol preparation of Example 1 is prepared by sodium pyruvate and baicalin-berberamine, while the lyophilized powder in the aerosol preparation of Comparative Example 1 is prepared by sodium pyruvate and baicalin, the lyophilized powder in the aerosol inhalation preparation of Comparative Example 2 is prepared by berbamine and sodium pyruvate, and the lyophilized powder in the aerosol preparation of Comparative Example 3 is prepared by a mixture of baicalin and berbamine and sodium pyruvate. In summary, baicalin-berberamine of the present invention has a strong effect of resisting Staphylococcus aureus and beta-hemolytic streptococci. That is, the sodium pyruvate aerosol inhalation preparation of the present invention has an excellent antibacterial effect.

[0051] Experimental Example 4 The pharmacological performance of the aerosol preparations of Example 1 and Comparative Examples 1-3 was investigated as follows: Twenty-five guinea pigs, weighing 290-320 g, were raised normally at 25°C, 60% humidity and 12-hour alternating light conditions. The guinea pigs were placed in a closed bell jar of a cough and asthma induction instrument in turn, and 0.6% histamine solution (solvent: normal saline) was atomized into the closed bell jar at an atomization rate of 0.15 mL / min using an ultrasonic nebulizer for 15 seconds. After stopping the atomization, the asthma induction latency of the guinea pigs (the time from the start of spraying to the onset of asthma symptoms such as dyspnea, convulsions and falls in the guinea pigs) was observed, and guinea pigs with a latency period of more than 2 minutes were discarded. 18 guinea pigs were screened and randomly divided into 6 groups, one of which was set as a blank control group.

[0052] 18 guinea pigs were anesthetized by intraperitoneal injection of urethane solution at a volume of 1.25 g / kg. After 30 minutes, the limbs and head of the guinea pigs were fixed, and jugular vein intubation and tracheal intubation were started. The respiratory flow rate, intrathoracic pressure, respiratory flow rate change and tidal volume of each group of guinea pigs were measured.

[0053] 0.5 mL of 15 μg / mL histamine was injected intravenously at a rate of 1 mL / min. After 30 minutes, each group of guinea pigs was given a nebulized inhalation of 0.8 mg / kg sodium pyruvate aerosol inhalation preparation for 2 minutes, the drug delivery device was removed, and the endotracheal tube was reconnected. After 5 minutes, 0.5 mL of 15 μg / mL histamine was injected intravenously. The guinea pigs were waited for to stabilize and the parameters of each group were recorded. After 4 hours of drug administration, 0.5 mL of 15 μg / mL histamine was injected intravenously. The guinea pigs were waited for to stabilize and the parameters of each group were recorded. Airway resistance = change in intrathoracic pressure / change in respiratory flow rate, dynamic lung compliance = tidal volume / change in intrathoracic pressure, where the change in inspiratory flow rate is not calculated based on the directly measured respiratory flow rate value, but the differential value of the tidal volume with respect to time (the size of the change in the volume of the tracing box over time) is the change in respiratory flow rate. This is because only when the gas actually enters the lungs can the gas exchange function be exerted and the chest movement be caused, thereby causing a change in tidal volume.

[0054] The airway resistance and dynamic lung compliance values ​​calculated each time the animal is in a stable state are taken as the normal values ​​before stimulation, and the values ​​with the largest changes in airway resistance and dynamic lung compliance after stimulation are taken as the airway resistance and dynamic lung compliance values ​​after administration. The calculation formula for the change rate is as follows: Airway resistance change rate = (maximum airway resistance after stimulation - normal airway resistance before stimulation) / normal airway resistance before stimulation; Dynamic lung compliance change rate = (minimum dynamic lung compliance after stimulation - normal dynamic lung compliance before stimulation) / normal dynamic lung compliance before stimulation. The experimental results of each group were averaged, and the results are shown in Table 4.

[0055] Table 4 As shown in Table 4, compared with the blank control group and comparative examples 1-3, the sodium pyruvate aerosol inhalation preparation of Example 1 has a significantly lower airway resistance increase rate and a significantly higher dynamic lung compliance decrease rate after administration, which indicates that the asthma of guinea pigs is significantly relieved after aerosolization, and the maintenance effect is better after a prolonged period of time. The above results show that the sodium pyruvate aerosol inhalation preparation of the present invention has excellent biological activity.

[0056] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit them. The basic principles and main features of the present invention have been described above with specific implementation schemes. On the basis of the present invention, some modifications or replacements may be made thereto, but these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of protection claimed by the present invention.

Claims

1. A sodium pyruvate aerosol inhalation preparation, characterized in that: The method comprises the following raw materials in parts by weight: 0.04-0.09 parts of baicalin-berbamine / sodium pyruvate inclusion compound freeze-dried powder, 100 parts of normal saline; The preparation process of baicalin-berberamine is as follows: Berbamine reacts with 4-chlorophenyl glycidyl ether to obtain intermediate A, and the intermediate A is mixed with baicalin solution and reacted under catalyst conditions. After the reaction is completed, baicalin-berbamine is purified to obtain baicalin-berbamine.

2. The sodium pyruvate aerosol inhalation preparation according to claim 1, characterized in that: The molar ratio of the intermediate A, baicalin and the catalyst is 1:0.25-0.5:15-20, and the catalyst is triethylamine.

3. The sodium pyruvate aerosol inhalation preparation according to claim 1, characterized in that: The conditions for the reaction of the intermediate A with the baicalin solution are: reacting at 75-90° C. for 30-60 min.

4. The sodium pyruvate aerosol inhalation preparation according to claim 1, characterized in that: The molar ratio of berbamine to 4-chlorophenyl glycidyl ether is 1:1.2-1.5, and the reaction conditions are: reacting at 55-70° C. for 2-4 hours.

5. The sodium pyruvate aerosol inhalation preparation according to claim 1, characterized in that: The preparation method of the baicalin-berbamine / sodium pyruvate inclusion compound freeze-dried powder comprises: using a polymer as a carrier, and adopting an emulsified solvent volatilization method to load sodium pyruvate and baicalin-berbamine onto the polymer to prepare nanoparticles; performing surface hydrophilic modification on the nanoparticles, and adding the modified nanoparticles into a freeze-drying protective agent solution, and freeze-drying to obtain the freeze-dried powder.

6. The sodium pyruvate aerosol inhalation preparation according to claim 5, characterized in that: The mass ratio of the sodium pyruvate, baicalin-berberamine and polymer is 1:1-10:1-10, and the polymer is selected from one of polylactic acid, polylactic acid-glycolic acid copolymer and polycaprolactone.

7. The sodium pyruvate aerosol inhalation preparation according to claim 5, characterized in that: The concentration of the hydrophilic compound used in the hydrophilic modification is 0.3-0.6 mol / L, the hydrophilic compound is selected from polyethylene glycol or polyvinyl pyrrolidone, and the lyophilization protective agent is composed of mannitol and trehalose in a mass ratio of 1:1-2.

8. The sodium pyruvate aerosol inhalation preparation according to claim 5, characterized in that: The freeze drying process is specifically as follows: first pre-freeze at normal pressure -30°C to -40°C for 2 to 4 hours; then evacuate and keep warm at 5 to 30 Pa and -1 to -10°C for 25 to 30 hours; then keep warm at 0 to 5°C for 1 to 2 hours; then heat to 25 to 30°C and keep warm for 12 to 16 hours.

9. The sodium pyruvate aerosol inhalation preparation according to claim 5, characterized in that: The specific operation of the emulsified solvent volatilization method includes: adding the oil phase to the water phase under stirring, emulsifying and solidifying, centrifuging and washing; the water phase is a polyvinyl alcohol solution, and the oil phase is selected from one of a polylactic acid solution, a polylactic acid-glycolic acid copolymer solution, and a polycaprolactone solution; Wherein, the mass concentration of the polyvinyl alcohol in the water phase is 0.8-1.5%.

10. A method for preparing a sodium pyruvate aerosol inhalation preparation, characterized in that: The baicalin-berbamine / sodium pyruvate inclusion compound freeze-dried powder is compounded with physiological saline according to the ratio described in claim 1 to obtain the product.

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