Cevimeline sodium suspensions, lyophilized powders, inhalation aerosols, and methods of making same

Abstract

The application discloses a kind of sodium cilevestat suspension, freeze-dried powder, atomization inhalant and preparation method thereof, the sodium cilevestat suspension is the mixed solution of containing osmotic pressure regulator and surfactant, sodium cilevestat and porous phospholipid particle are dispersed after shearing stirring, then homogenization is obtained;The mass ratio of sodium cilevestat and porous phospholipid particle is 1.2~2:1.The sodium cilevestat suspension system of the application is stable, uniformly dispersed for a long time, after preparation by spray freeze-drying, sodium cilevestat freeze-dried powder is redissolved, can still maintain suspension state, and the dispersion performance is excellent;After atomization by atomization device, the percentage of particle diameter below 5.0 μm is high, fine particle fraction FPF is more than 70%, with excellent aerodynamic characteristics, more particles are deposited in 4~5 grade, more suitable for deep lung alveoli and lung microvessels, for the treatment of acute lung injury / acute respiratory distress syndrome.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical preparation technology, and more specifically, this invention relates to a sodium cevelestat suspension, lyophilized powder, nebulized inhaler, and its preparation method. Background Technology

[0002] ALI / ARDS refers to acute, progressive hypoxic respiratory failure caused by various intrapulmonary and extrapulmonary pathogenic factors other than cardiac causes. ALI is a serious clinical condition that can develop into ARDS. The main pathological features of ALI / ARDS are increased pulmonary microvascular permeability leading to protein-rich pulmonary edema and hyaline membrane formation in the alveolar exudate, accompanied by pulmonary interstitial fibrosis. Clinically, it presents as refractory hypoxemia, respiratory rate, and respiratory distress. Chest X-ray shows diffuse infiltrates in both lungs, and in later stages, multiple organ dysfunction often occurs.

[0003] Cevelexastat sodium is the only drug globally used for ALI / ARDS. As a highly specific neutrophil elastase inhibitor, it directly inhibits the core of the inflammatory response. Neutrophil elastase is a proteolytic enzyme that, when released from neutrophils aggregated in the lungs, can break down lung connective tissue, increase pulmonary vascular permeability, and induce ALIARDS. It can also promote the production of neutrophil chemokines, exacerbating the inflammatory response, and is an important damaging factor associated with ALIARDS and systemic inflammatory response syndrome. Clinical studies have shown that cevelexastat sodium can effectively reduce lung injury scores, improve lung function, shorten mechanical ventilation time and intensive care unit hospital stay in ARDS patients, and reduce mortality.

[0004] Currently, cevelexat sodium is only available as an injection, requiring continuous intravenous administration over 24 hours. This can easily lead to adverse reactions such as abnormal liver function and poses a potential risk of inducing systemic toxicity. Developing cevelexat sodium into an inhalation formulation increases the local concentration of the drug in the lungs. Through the dense capillary network of the lungs, drug absorption is accelerated, thereby reducing its impact on other tissues throughout the body, making it the preferred method of administration for treating lung diseases. However, cevelexat sodium has poor water solubility and requires extremely high clinical doses. Therefore, a solvent-free nebulized inhaler is the preferred dosage form for developing cevelexat sodium into an inhalation formulation. For example, CN116115589A discloses a pharmaceutical composition of cevelexta sodium for nebulized inhalation. Cevelexta sodium and the amphiphilic material distearylphosphatidylethanolamine-polyethylene glycol are dissolved separately in organic solvents, mixed in a certain proportion, and the organic solvent is removed by vacuum evaporation to obtain a drug-containing lipid film. After dissolving in water, a certain amount of mannitol is added, and the mixture is lyophilized. Before use, the mixture is first reconstituted with water and then nebulized through an inhaler. CN116635018A discloses a lyophilized powder of cevelexta sodium for nebulized inhalation, containing the excipient mannitol, a pH adjuster, an osmotic pressure adjuster, and a surfactant. The solution is prepared at low temperature and then lyophilized. Before use, the lyophilized powder is dissolved in water or a carrier and nebulized using a conventional inhalation device. CN117982414B discloses sodium cevelestat nanocrystals, their dry powder, inhaled powder, and their preparation method. Using vitamin E polyethylene glycol succinate (TPGS) as a stabilizer, it is homogenized with an aqueous dispersion of sodium cevelestat under high pressure to prepare a nanocrystal suspension. The suspension is then solidified using spray freeze-drying technology to obtain sodium cevelestat nanocrystal dry powder, which is then encapsulated.

[0005] However, the current nebulized inhalation formulation of cevelexostat sodium still has the following problems: First, the cevelexostat sodium suspension has a short uniform dispersion time and is prone to sedimentation after shaking. After being made into a lyophilized powder, its redispersibility is poor, resulting in poor droplet uniformity after nebulization, which affects its dispersion and deposition during inhalation. Second, the percentage of particles smaller than 5μm (fine particle fraction, FPF) in the suspension is low. When used for the treatment of ALI / ARDS, its aerodynamic properties are insufficient, and more particles are deposited at levels 2-3, failing to penetrate deep into the alveoli and pulmonary microvessels. Third, the drug is released rapidly in the body, requiring continuous administration, and there is also a potential risk of toxicity. Summary of the Invention

[0006] Based on this, the purpose of the present invention is to provide a sodium cevelestat suspension with a long uniform dispersion time. The lyophilized powder prepared using the suspension has good redispersibility, a high percentage of fine particles in the prepared nebulized inhaler, excellent aerodynamic characteristics, and a moderate drug release rate.

[0007] The specific technical solutions for achieving the above-mentioned objectives are as follows.

[0008] In a first aspect, the present invention provides a sodium cevelestat suspension, which is obtained by dispersing a mixed solution containing an osmotic pressure regulator and a surfactant with sodium cevelestat and porous phospholipid particles by shearing and stirring, followed by homogenization; wherein the mass ratio of sodium cevelestat to porous phospholipid particles is 1.2 to 2:1.

[0009] In a second aspect, the present invention provides a method for preparing the above-mentioned cevelexat sodium suspension, comprising the following steps: adding cevelexat sodium and porous phospholipid particles to a mixed solution containing an osmotic pressure regulator and a surfactant, dispersing by shearing and stirring, and then homogenizing.

[0010] In a third aspect, the present invention provides a lyophilized sodium cevelexta powder, which is obtained by spray-freezing the above-mentioned sodium cevelexta suspension.

[0011] In a fourth aspect, the present invention provides a cevelexta sodium nebulized inhaler comprising the above-mentioned cevelexta sodium lyophilized powder.

[0012] This invention utilizes lightweight porous phospholipid particles as a carrier to load cevelexat sodium drug crystals, and then disperses them by shearing and stirring with a mixed solution containing an osmotic pressure regulator and a surfactant, followed by homogenization, to prepare a stable cevelexat sodium suspension with a uniform dispersion time exceeding 5 minutes, and in some cases exceeding 20 minutes. This cevelexat sodium suspension is then spray-dried to obtain cevelexat sodium lyophilized powder, which is then formulated into a nebulized inhaler. After reconstitution, the lyophilized powder remains in suspension and does not settle for at least 24 hours, exhibiting excellent redispersibility. When nebulized, the fine particle fraction (FPF) (percentage of particles smaller than 5.0 μm) exceeds 70%, exhibiting excellent aerodynamic characteristics. A large number of particles deposit at level 4-5, making it more suitable for deep penetration into the alveoli and pulmonary microvessels, and is used for the treatment of acute lung injury (ALI) / acute respiratory distress syndrome (ARDS).

[0013] This invention controls the ratio of cevelexostat sodium to light porous phospholipid particles within a certain range, and combines this with appropriate shear dispersion and homogenization processes (especially microfluidic homogenization) to prepare a cevelexostat sodium suspension with a low PDI, high Zata potential, high stability, and long uniform dispersion time. When made into an nebulized inhaler, it has a suitable particle size and a moderate drug release rate, thus overcoming the problem of the need for continuous administration and the potential risk of toxicity. Attached Figure Description

[0014] Figure 1The results show the in vitro release rate of the cevelexostat sodium suspension in Experiment Example 6 of this invention.

[0015] Figure 2 The deposition results of cevelexatana at each layer are shown in Experimental Example 8 of this invention. Detailed Implementation

[0016] To facilitate understanding of the present invention, a more complete description will be provided below. The present invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a thorough and complete understanding of the disclosure of the present invention.

[0017] Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used in this specification is for the purpose of describing particular embodiments only and is not intended to limit the invention. The term "and / or" as used in this invention includes any and all combinations of one or more of the associated listed items.

[0018] Unless otherwise specified, all examples are conducted under conventional experimental conditions or as recommended in the manufacturer's instructions. All raw materials and reagents used in this invention are commercially available.

[0019] In some embodiments of the present invention, a sodium cevelestat suspension is disclosed, which is obtained by dispersing a mixed solution containing an osmotic pressure regulator and a surfactant with sodium cevelestat and porous phospholipid particles by shearing and stirring, and then homogenizing; the mass ratio of sodium cevelestat to porous phospholipid particles is 1.2 to 2:1.

[0020] In some embodiments, the mass ratio of sodium cevelestat to porous phospholipid particles is 1.2 to 1.5:1, more preferably 1.2 to 1.4:1.

[0021] In some embodiments, the pH of the mixed solution is 6.5 to 8.5, preferably 6.5 to 7.5, and more preferably 6.5 to 7.0.

[0022] In some embodiments, the osmotic pressure regulator is sodium chloride and / or glucose, preferably sodium chloride.

[0023] In some embodiments, the surfactant is sodium dodecyl sulfate and / or Tween 80, preferably Tween 80.

[0024] In some embodiments, the concentration of sodium chloride in the mixed solution is 8 mg / mL to 10 mg / mL, preferably 8.5 mg / mL to 9.5 mg / mL, and the concentration of Tween 80 is 0.01 mg / mL to 10.0 mg / mL, preferably 0.01 mg / mL to 5.0 mg / mL, more preferably 1.5 mg / mL to 1.7 mg / mL.

[0025] In some embodiments, the mass-to-volume ratio of the sodium cefilestat to the mixed solution is 40 mg to 60 mg: 1 mL, preferably 50 mg to 60 mg: 1 mL, and more preferably 55 mg to 60 mg: 1 mL.

[0026] In some embodiments, the shearing and stirring speed is 10,000 rpm to 12,000 rpm, preferably 10,000 rpm to 11,000 rpm, and more preferably 10,000 rpm to 10,050 rpm.

[0027] In some embodiments, the shearing and stirring time is 10 min to 15 min, preferably 10 min to 13 min, and more preferably 10 min to 11 min.

[0028] In some embodiments, the homogenization is high-pressure homogenization or microfluidic homogenization, more preferably microfluidic homogenization. Civelecustat sodium and porous phospholipid particles are passed through a Y-shaped homogenization chamber of a microfluidic homogenizer, ensuring effective adsorption of the drug onto the surface and interior of the porous phospholipid particles, achieving high-efficiency loading. Homogenization is then achieved through the high shear force and turbulence of a Z-shaped homogenization chamber.

[0029] In some embodiments, the homogenization pressure of the microjet homogenization is 15,000 psi to 20,000 psi, preferably 18,000 psi to 20,000 psi, and more preferably 19,000 psi to 20,000 psi.

[0030] In some embodiments, the microjet homogenization is performed 2 to 3 times, more preferably 2 times.

[0031] In some embodiments, the d90 particle size of the porous phospholipid particles is 2 μm to 4 μm.

[0032] In some embodiments, the porous phospholipid particles have a pore size of 0.2 μm to 1 μm.

[0033] In some embodiments, the density of the porous phospholipid particles is 0.4 g / cm³. 3 ~0.5g / cm 3 .

[0034] In some embodiments, the porous phospholipid particles are prepared by the following method: DSPC and CaCl2 are added to water for injection at a molar ratio of 1 to 3:1 and dispersed, while a pore-forming agent is added with stirring, followed by shearing and stirring, high-pressure homogenization, and then spray drying.

[0035] In some embodiments, the molar ratio of DSPC to CaCl2 is 1.5 to 2.5:1, more preferably 1.8 to 2.2:1.

[0036] In some embodiments, the mass ratio of DSPC and CaCl2 to the volume ratio of water for injection is 3-15 g:100 mL, preferably 3.6-13.5 g:100 mL, more preferably 4-6 g:100 mL, and even more preferably 4.5-5.5 g:100 mL.

[0037] In some embodiments, the porogen is perfluorobromooctane (PFOB) or perfluoropropane, preferably PFOB.

[0038] In some embodiments, the volume ratio of water for injection to pore-forming agent is 1 to 4:1, preferably 2 to 4:1, and more preferably 3 to 4:1.

[0039] In some embodiments, the shearing and stirring speed is 8000 rpm to 10000 rpm.

[0040] In some embodiments, the pressure of the high-pressure homogenization is 140MPa to 180MPa, preferably 150MPa to 170MPa, more preferably 160MPa to 170MPa; the number of high-pressure homogenizations is 3 to 6 times, preferably 3 to 5 times, more preferably 4 to 5 times.

[0041] In some embodiments, the process parameters of the spray drying include: inlet temperature 90℃~135℃; outlet temperature 68℃~72℃; feed rate 2mL / min~8mL / min; and total gas flow rate 425L / min~525L / min.

[0042] In other embodiments of the present invention, a method for preparing the above-mentioned cevelexat sodium suspension is disclosed, comprising the following steps: adding cevelexat sodium and porous phospholipid particles to a mixed solution containing an osmotic pressure regulator and a surfactant, dispersing by shearing and stirring, and then homogenizing.

[0043] In some embodiments, the shearing and stirring speed is 10,000 rpm to 12,000 rpm, preferably 10,000 rpm to 11,000 rpm, and more preferably 10,000 rpm to 10,050 rpm.

[0044] In some embodiments, the shearing and stirring time is 10 min to 15 min, preferably 10 min to 13 min, and more preferably 10 min to 11 min.

[0045] In some embodiments, the homogenization is high-pressure homogenization or microjets homogenization.

[0046] In some embodiments, the homogenization pressure of the microjet homogenization is 15,000 psi to 20,000 psi, preferably 18,000 psi to 20,000 psi, and more preferably 19,000 psi to 20,000 psi.

[0047] In some embodiments, the high-pressure homogenization pressure is 1000 bar to 1400 bar, and the high-pressure homogenization time is 10 min to 30 min; preferably, the high-pressure homogenization pressure is 1100 bar to 1300 bar, and the high-pressure homogenization time is 10 min to 20 min; more preferably, the high-pressure homogenization pressure is 1150 bar to 1250 bar, and the high-pressure homogenization time is 10 min to 15 min.

[0048] In some other embodiments of the present invention, a lyophilized sodium cevelexate is disclosed, which is obtained by spray freeze-drying the above-mentioned sodium cevelexate suspension.

[0049] In other embodiments of the present invention, a cevelexat sodium nebulized inhaler is disclosed, comprising the above-mentioned cevelexat sodium lyophilized powder. The cevelexat sodium nebulized inhaler is mixed with water for injection, nebulized through a nebulizer (preferably a vibrating mesh nebulizer), and inhaled. Based on existing daily dosages for injectable drugs, the dosage is 5 mL, and the drug strength is 60 mg / mL.

[0050] The porous phospholipid particles for loading cevelexatol sodium described in this invention can be prepared according to the suspension particle preparation method in the prior art (CN105193773 B). In the following examples, the specific preparation steps of the porous phospholipid particles are as follows: 18.7 g of DSPC (1,2-distearate-sn-glycerol-3-phosphocholine) and 1.3 g of calcium chloride were homogenized in 400 mL of hot water (75°C) using a high-shear stirrer (8000 rpm); 100 mL of PFOB was slowly added during homogenization; the crude emulsion was further homogenized 5 times using a high-pressure homogenizer at a pressure of 170 MPa; the emulsion was spray-dried under nitrogen, with an inlet temperature of 95°C, an outlet temperature of 72°C, an emulsion feed rate of 2.4 mL / min, and a total gas flow rate of 498 L / min. The prepared porous phospholipid particles have a d90 particle size of approximately 2 μm, a pore size of 0.5 μm, and a density of 0.5 g / cm³.3 The density of the loaded cevelexat sodium is close to 1.0 g / cm³. 3 This ensures good suspension performance and prevents it from settling.

[0051] The microfluidic homogenizer used in this invention is manufactured by Norzer Fluid Technology (Shanghai) Co., Ltd., and its model is nano-LA 75Y.

[0052] The present invention will be further described in detail below with reference to specific embodiments.

[0053] Example 1: Cevelexostat sodium suspension, lyophilized powder

[0054] This embodiment first provides a cevelexostat sodium phospholipid complex suspension, which is prepared through the following steps:

[0055] (1) Sodium chloride and Tween 80 were added to 1 mL of water for injection at concentrations of 9 mg / mL and 1.6 mg / mL, respectively. The mixture was stirred until dissolved. The pH of the solution was adjusted to 7.5 using a buffer solution (pH 12.5) made of sodium dihydrogen phosphate and sodium hydroxide.

[0056] (2) Add 60 mg of cevelexatol sodium and 46.15 mg of porous phospholipid particles to the above solution at a mass ratio of 1.3:1, and mechanically stir for 10 min at a speed of 10000 rpm to disperse them initially.

[0057] (3) Then, a high-pressure homogenizer was used to homogenize the solution at 1200 bar for 15 min to prepare a sodium ceftriaxone suspension with a concentration of 60 mg / mL.

[0058] In this embodiment, the prepared cevelexostatin suspension was converted into cevelexostatin lyophilized powder using a spray freeze-drying method, as follows:

[0059] Cevelexostat sodium suspension was atomized using a 0.7 mm dual-fluid nozzle and dispersed in liquid nitrogen for rapid freezing to form small ice crystals. During dispersion, the liquid nitrogen was continuously stirred at a speed of 500 rpm / min. The small ice crystals were then transferred to a freeze dryer (Booqi Lyovapor™ L-200pro) for spray freeze-drying for 48 hours. The spray freezing parameters were: compressed air flow rate 473 L / h, feed rate 20% (approximately 4 mL / min), spray pressure 5 bar, and freezing vacuum 0.4 mbar.

[0060] Example 2: Cevelexta sodium suspension, lyophilized powder

[0061] This embodiment first provides a cevelexostat sodium phospholipid complex suspension, which is prepared through the following steps:

[0062] (1)~(2) Same as Example 1;

[0063] (3) Using the Y-type interactive cavity or Z-type auxiliary cavity of the microfluidic homogenizer, homogenize twice under the conditions of homogenization pressure of 20000psi, homogenization speed of 53mm / s and condensation circulation temperature of 25℃ to prepare a sodium cevelestat suspension with a concentration of 60mg / mL.

[0064] In this embodiment, the cevelextanas suspension was converted into cevelextanas lyophilized powder by spray freeze-drying, the same method as in Example 1.

[0065] Comparative Example 1: Inhalation of Civelestat Sodium Suspension

[0066] This comparative example provides an inhaled cefelesat sodium suspension, the preparation method of which is as follows:

[0067] 1. Weigh out Tween 80, sorbitan laureate, and NaCl, add water for injection and stir to dissolve, and prepare mixed solutions with concentrations of 14 mg / mL, 1.8 mg / mL, and 9 mg / mL respectively. Then adjust the pH of the above solution to 7.5 with a buffer solution made of sodium dihydrogen phosphate and sodium hydroxide (pH value of 12.5), add cevelexat sodium and stir to mix evenly.

[0068] 2. A high-pressure homogenizer was used, with a homogenization pressure of 1000 bar and a feed rate of 120 L / h, to homogenize to the desired particle size (X50 = 1.38; X90 = 2.41 μm), thus obtaining a 60 mg / mL cevelexta sodium suspension. Comparative Example 2: Cevelexta sodium nanocrystal suspension for inhalation.

[0069] This comparative example provides an inhaled cefelesstat sodium nanocrystal suspension, the preparation method of which is as follows:

[0070] 1. Dissolve cetirizine sodium in water for injection to prepare a solution with a concentration of 60 mg / mL. Adjust the pH of the solution to 7.5 with sodium dihydrogen phosphate-sodium hydroxide buffer (pH 12.5). Then add sodium chloride and Tween 80 to final concentrations of 9 mg / mL and 1 mg / mL, respectively.

[0071] 2. Use a high-shear stirrer at 12000 rpm for 10 minutes to perform preliminary dispersion and obtain a dispersion liquid;

[0072] 3. The obtained dispersion was homogenized in a high-pressure homogenizer at a homogenization pressure of 1500 bar for 15 min to obtain a sodium cevelestat nanocrystal suspension with a concentration of 60 mg / mL.

[0073] Comparative Example 3: Inhalation of Civelestat Sodium Nanocrystal Suspension

[0074] This comparative example provides an inhaled cefelesstat sodium nanocrystal suspension, the preparation method of which is as follows:

[0075] 1. Dissolve cetirizine sodium in water for injection to prepare a solution with a concentration of 60 mg / mL. Adjust the pH to 7.5 with sodium dihydrogen phosphate-sodium hydroxide buffer. Add sodium chloride and Tween 80 to final concentrations of 9 mg / mL and 1 mg / mL, respectively.

[0076] 2. Use a high-shear stirrer at 12000 rpm for 10 minutes to perform preliminary dispersion and obtain a dispersion liquid;

[0077] 3. Using the Y-type interactive cavity or Z-type auxiliary cavity of the microfluidic homogenizer, the obtained dispersion is homogenized twice under the conditions of homogenization pressure of 20000psi, homogenization speed of 53mm / s, and condensation circulation temperature of 25℃ to obtain a sodium cevelestat nanocrystal suspension with a concentration of 60mg / mL.

[0078] Comparative Example 4: Inhalation of Cefelestat Sodium Liposome Suspension

[0079] This comparative example uses a thin-film dispersion method to prepare an inhaled cevelexostat sodium liposome suspension, the preparation method of which is as follows:

[0080] 1. Weigh 1.44g DSPC, 0.36g cholesterol, and 0.03g α-tocopherol and dissolve them in chloroform. Weigh 1.8g cevelexostat sodium and add it to an appropriate amount of methanol. Stir until dissolved. Combine the solutions and evaporate under low temperature and reduced pressure to remove the organic solvent, thus obtaining a phospholipid membrane.

[0081] 2. Add 0.27g of sodium chloride and 0.48g of Tween 80 to a sodium dihydrogen phosphate-sodium hydroxide buffer solution with a pH of 7.5 to obtain an aqueous phase. Slowly add the aqueous phase to the phospholipid membrane, gently shaking the flask to allow the phospholipid membrane to mix and hydrate better with the aqueous phase, thus obtaining an emulsion;

[0082] 3. Using the Y-type interactive cavity or Z-type auxiliary cavity of the microfluidic homogenizer, the obtained emulsion is homogenized twice under the conditions of homogenization pressure of 15000psi, homogenization speed of 53mm / s, and condensation circulation temperature of 25℃.

[0083] 4. Remove organic solvents by purging with nitrogen gas, and then use a 0.9% NaCl solution through a tangential flow ultrafiltration system (hollow fiber column membrane area 118 cm²). 2 Wash and filter (fiber inner diameter 0.5mm) to remove free drug, and concentrate the volume to 30mL;

[0084] 5. Sterilize and filter using a sterile 0.22μm membrane filter, and refrigerate at 4℃ for 2 hours to obtain a 60mg / mL cevelexat sodium liposome suspension.

[0085] Comparative Example 5: Inhalation of Civelestat Sodium Liposome Suspension

[0086] This comparative example uses an emulsification-solidification method to prepare an inhalation-grade cevelexostat sodium liposome suspension, the preparation method of which is as follows:

[0087] 1. Weigh 1.44g DSPC, 0.36g cholesterol, and 0.03g α-tocopherol and dissolve them in chloroform. Weigh 1.8g cevelexat sodium and add it to an appropriate amount of methanol. Stir until dissolved. Combine the solutions to obtain an organic phase solution.

[0088] 2. Add 0.27g sodium chloride and 0.48g Tween 80 to water for injection in sequence, stir to dissolve, and adjust the pH of the above solution to 7.5 with sodium dihydrogen phosphate-sodium hydroxide buffer (pH 12.5) to obtain the aqueous phase;

[0089] 3. In a 45°C water bath, and under continuous magnetic stirring, the organic phase is slowly dripped into the aqueous phase to form an O / W emulsion;

[0090] 4. Using the Y-type interactive cavity or Z-type auxiliary cavity of the microfluidic homogenizer, the obtained emulsion is homogenized twice under the conditions of homogenization pressure of 15000psi, homogenization speed of 53mm / s, and condensation circulation temperature of 25℃.

[0091] 5. Remove organic solvents by purging with nitrogen gas, and then use a 0.9% NaCl solution through a tangential flow ultrafiltration system (hollow fiber column membrane area 118 cm²). 2 Wash and filter (fiber inner diameter 0.5mm) to remove free drug, and concentrate the volume to 30mL;

[0092] 6. Sterilize and filter using a sterile 0.22μm membrane filter, and refrigerate at 4℃ for 2 hours to obtain a 60mg / mL cevelexat sodium liposome suspension.

[0093] Experimental Example 1: Investigation of the suspension properties of cevelexta sodium suspension

[0094] 1. Sedimentation and stratification: The suspension samples of Examples 1-2 and Comparative Examples 1-5 were shaken for 30 seconds and then allowed to stand to observe the sedimentation and stratification of the samples and the time.

[0095] 2. Average particle size, polydispersity index (PDI), and zeta potential: Take the suspension samples from Examples 1-2 and Comparative Examples 1-5, dilute them 100 times with pure water until clear and transparent, pipette 1 mL into a quartz cuvette, place it in a Malvern nanoparticle size analyzer, and determine the particle size, polydispersity index (PDI), and zeta potential of the cevelexostat sodium suspension using dynamic light scattering method. Perform the measurements in parallel for 3 times and take the average value.

[0096] The results are shown in Table 1.

[0097] Table 1. Suspension characteristics and particle size results of cevelexta sodium suspension.

[0098]

[0099] As shown in Table 1, the sodium celestatin suspension prepared by conventional methods (Comparative Example 1) is prone to sedimentation, with a PDI > 0.3 and a small absolute value of the Zeta potential (less than 30 mV), indicating instability. The sodium celestatin nanocrystal suspensions (Comparative Examples 2 and 3), sodium celestatin liposome suspensions (Comparative Examples 4 and 5), and sodium celestatin phospholipid complex suspensions (Examples 1 and 2) all maintained a uniform dispersion state for ≥3 min. Among these, the sodium celestatin suspensions (Examples 1 and 2) loaded with porous phospholipid particles as a carrier were uniformly dispersed in the aqueous solution for more than 20 min after shaking, significantly exceeding the dispersion time of the sodium celestatin nanocrystal suspensions (Comparative Examples 2 and 3) and the sodium celestatin liposome suspensions (Comparative Examples 4 and 5).

[0100] Except for Comparative Example 1, the PDI of suspensions in different forms (nanocrysts, liposomes, and phospholipid complexes) remained below 0.3, and the absolute value of the Zeta potential was above 30 mV, indicating system stability. However, the particle size differences were significant. The particle size of the cevelexostat sodium phospholipid complex suspension (Examples 1 and 2) was significantly larger than that of the cevelexostat sodium nanocrystal suspension (Comparative Examples 2 and 3) and the cevelexostat sodium liposome suspension (Comparative Examples 4 and 5). Smaller drug particle size results in a larger specific surface area, leading to faster drug release in vivo, requiring continuous administration, and posing a potential risk of toxicity. Therefore, the cevelexostat sodium phospholipid complex suspension better meets the medication requirements.

[0101] Experimental Example 2: Redispersibility of Cevelexta Sodium Lyophilized Powder

[0102] Accurately weigh 0.3 g of lyophilized sodium cevelexate from Examples 1-2 and Comparative Examples 2-5 (preparation process as in Example 1) and disperse it in 10 mL of pure water. Examine the changes in particle size, PDI, and Zeta potential of the lyophilized sodium cevelexate after redispersibility. Calculate the redispersibility index according to formula (2). The results are shown in Table 2.

[0103]

[0104] Table 2 Results of redispersibility after reconstitution of cevelexta sodium lyophilized powder

[0105] No. Average particle size (nm) PDI Zeta potential (mV) Rindex Example 1 2506 0.19 -42.29 1.05 Example 2 2108 0.17 -43.81 1.03 Comparative Example 2 962 0.27 -38.27 1.43 Comparative Example 3 695 0.28 -32.45 1.67 Comparative Example 4 283 0.26 -39.24 1.27 Comparative Example 5 252 0.23 -38.63 1.09

[0106] A redispersibility index closer to 1 indicates better redispersibility performance. A particle PDI less than 0.3 and an absolute value of the Zeta potential greater than 30 indicate more stable particles. Table 2 shows that all lyophilized powders maintained good stability after redispersibility. However, the redispersibility performance of the lyophilized powders prepared from the cevelexostat sodium phospholipid complex suspension (Examples 1 and 2) was significantly better than that of the lyophilized powders prepared from the cevelexostat sodium liposome suspension (Comparative Examples 4 and 5) and the lyophilized powders prepared from the cevelexostat sodium nanocrystal suspension (Comparative Examples 2 and 3). Furthermore, compared to the cevelexostat sodium phospholipid complex suspension prepared by high-pressure homogenization (Example 1), the cevelexostat sodium phospholipid complex suspension prepared by microfluidic homogenization (Example 2) had smaller particle size, better stability, and superior dispersibility.

[0107] Experimental Example 3: Comparison of the effect of the ratio of drug to porous phospholipid particles on the sodium cevelestat suspension.

[0108] Cevelexostat sodium suspension was prepared according to the method of Example 2, except that in step (2), the mass ratio of cevelexostat sodium to porous phospholipid particles was 2:1, 1.4:1, 1.3:1, 1.2:1, 1:1, and 1:2, respectively. The suspension properties of each suspension were investigated according to the method of Experimental Example 1, and the results are shown in Table 3.

[0109] Table 3. Dispersion state, particle size, PDI, and Zeta potential of each suspension.

[0110]

[0111] The mass ratio of sodium cevelestat to porous phospholipid particles has a significant impact on the suspension state of the suspension. If the proportion of sodium cevelestat is too high, sedimentation and stratification are likely to occur. If the proportion of porous phospholipid particles is too high, stratification and floating are likely to occur. As shown in Table 3, when the mass ratio of sodium cevelestat to porous phospholipid particles is 1:1 or 1:2, the PDI > 0.3 or the absolute value of the Zeta potential is less than 30 mV, the suspension is unstable, the uniform dispersion time is short, and the dispersion performance is poor. When the mass ratio of sodium cevelestat to carrier particles exceeds 1:1, after shaking for 30 s and standing, the uniform dispersion time is ≥ 3 min, and the PDI ≤ 0.3 and the absolute value of the Zeta potential is greater than 30 mV, which can achieve a stable state of the suspension. Among them, when the mass ratio of sodium cevelestat to porous phospholipid particles is 1.3:1, the prepared suspension has the longest uniform dispersion time and the smallest particle size, while the PDI is lower than 0.3 and the absolute value of the Zeta potential is larger.

[0112] Comparison of the effects of preparation process on cevelexostat sodium suspension in Experiment Example 4

[0113] The cevelexta sodium suspension was prepared according to the method of Example 2, except that different dispersion or homogenization processes were used in step (2) or step (3):

[0114] Group 1: In step (2), the stirring speeds are 8000 rpm, 10000 rpm, and 12000 rpm, respectively, and the other steps are the same as in Example 1.

[0115] Group 2: In step (2), the stirring time is 5 min, 10 min, and 15 min respectively, and the other steps are the same as in Example 1.

[0116] Group 3: In step (3), the microjet homogenization pressures are 15000psi, 20000psi, 25000psi and 30000psi respectively, and the other steps are the same as in Example 2.

[0117] Group 4: In step (3), the number of times the microjet homogenizes is 1, 2, and 3, respectively, and the other steps are the same as in Example 2.

[0118] The suspension properties of each suspension were examined according to the method in Experiment Example 1, and the results are shown in Table 4.

[0119] Table 4. Effects of different dispersion and homogenization processes on cevelexta sodium suspension

[0120]

[0121]

[0122] Table 4 shows that the high-shear stirring speed and time, as well as the microjets homogenizing pressure and the number of homogenization cycles, all significantly affect the homogenity of the cevelex suspension. A stable suspension can only be achieved when the high-shear stirrer speed exceeds 10,000 rpm, the stirring time exceeds 10 min, the microjets homogenizing pressure is below 20,000 psi, and the number of homogenization cycles is greater than 2. The PDI is lowest under the conditions of a stirring speed of 10,000 rpm and a stirring time of 10 min. When the microjets homogenizing pressure is 20,000 psi and homogenization is performed twice, the cevelex sodium suspension has a moderate particle size, a PDI < 0.3, and a relatively large absolute value of the Zeta potential; the cevelex suspension prepared under these process conditions is the most stable.

[0123] Experimental Example 5: Comparison of the effects of pH on cevelexta sodium suspension

[0124] The cevelexta sodium suspension was prepared according to the method of Example 2, except that in step (1), the pH of the solution was adjusted to 6.5, 7.0, 7.5, 8.0, and 8.5 respectively using a buffer solution (pH value of 12.5) made of sodium dihydrogen phosphate and sodium hydroxide.

[0125] Considering that cetelestat sodium is easily hydrolyzed, and that pH value has a significant impact on the stability of cetelestat sodium nebulized inhaler, to ensure stability during the lyophilization process and use, cetelestat sodium suspensions prepared at different pH values ​​were placed for different time periods for related substance detection. Quantitative analysis was performed using HPLC with a Diamonsil C18 column (200 mm × 4.6 mm, 5 μm), a mobile phase of 0.02 mol / L potassium dihydrogen phosphate buffer solution (pH = 3.5)-acetonitrile (55:45), and a detection wavelength of 208 nm.

[0126] The temperature of the water for injection was controlled at 20℃, and the cevelexta sodium suspension was placed at 20℃ for 24 hours. The changes in impurity A (impurities generated by hydrolysis) and total impurities in the cevelexta sodium suspension were investigated at 0, 0.5h, 1h, 2h, 4h, 6h, 12h, and 24h. The results are shown in Table 5.

[0127] Table 5. Changes in relevant substances in cevelexostat sodium suspension over 24 hours.

[0128]

[0129] As shown in Table 5, the impurities A and total impurities of the cevelexostat sodium suspension gradually increased during the storage process. When the pH of the solution was between 6.5 and 7.5, the growth of impurities A and total impurities in the prepared cevelexostat sodium suspension was relatively slow and stable.

[0130] Experimental Example 6: In vitro release study of cevelexta sodium suspension

[0131] The in vitro release of cevelexostat sodium suspension was investigated using a flow-through cell and dialysis bag. The in vitro dissolution rate of the active pharmaceutical ingredient (API) in lyophilized powders prepared from cevelexostat sodium suspensions of Examples 1-2 and Comparative Examples 2-5 (prepared using the same method as in Example 1) was investigated in artificial simulated lung fluid. The composition of the artificial lung fluid included: MgCl2·6H2O (0.20 g / L), NaCl (6.02 g / L), KCl (0.30 g / L), Na2HPO4·12H2O (0.36 g / L), Na2SO4 (0.07 g / L), CaCl2 (0.28 g / L), CH3COONa (0.57 g / L), NaHCO3 (2.60 g / L), Na3H5C6O7·2H2O (0.10 g / L), and dipalmitoylphosphatidylcholine (0.02% w / v).

[0132] Three portions of lyophilized powder equivalent to 0.3 g of cefelestatin sodium were accurately weighed and reconstituted with 5 mL of water for injection. The solutions were then transferred to 3000 Da (cellulose ester) dialysis bags and placed in a flow cell. 200 mL of artificial lung fluid was added, and the drug release was investigated using a closed-loop method at 37°C. 1 mL of the solution was collected at 0, 1 min, 5 min, 10 min, 15 min, 20 min, 30 min, 60 min, 120 min, 240 min, and 360 min, and 1 mL of artificial lung fluid was added to the release medium. The cefelestatin sodium content was determined by HPLC (parameters same as in Example 5).

[0133] The results are as follows Figure 1 As shown. From Figure 1 It can be seen that in the artificial lung fluid, sodium cetylestradiol sodium nanocrystal suspension (Comparative Examples 2-3), sodium cetylestradiol sodium liposome suspension (Comparative Examples 4-5), and sodium cetylestradiol sodium phospholipid complex suspension (Examples 1-2) can all gradually release sodium cetylestradiol sodium. Among them, sodium cetylestradiol sodium nanocrystal suspension has the fastest release rate, releasing more than 90% of the drug in about 30 minutes (Comparative Examples 2-3). Sodium cetylestradiol sodium liposome suspension reaches a release rate of more than 90% within 6 hours, which is relatively sufficient (Comparative Examples 4-5). Sodium cetylestradiol sodium phospholipid complex suspension has a moderate drug release rate, reaching a release rate of 90% in 2 hours (Examples 1-2).

[0134] Experimental Example 8: Investigation of the atomization characteristics of cevelexta sodium lyophilized powder

[0135] 1. Total delivery volume and delivery efficiency

[0136] The lyophilized sodium cefelestatin samples from Examples 1-2 and Comparative Examples 2-5 (prepared using the same method as in Example 1) were reconstituted with water for injection and nebulized using a PARI eflow nebulizer, following the procedure outlined in General Chapter 0111 of Part IV of the 2020 Chinese Pharmacopoeia. The nebulizer was connected, and the breathing simulator was set to adult mode. The first nebulization lasted 60 seconds. The first filter paper was removed, and the second filter paper was replaced before nebulization continued for 5 minutes. All filter papers (first, second, exhaled, nebulizer cup, filter paper assembly, and adapter) were thoroughly washed with methanol. The active substance was quantitatively collected, with three parallel runs for each nebulization time. Quantitative analysis was performed using HPLC. The chromatographic column was a Diamonsil C18 (200 mm × 4.6 mm, 5 μm), the mobile phase was 0.02 mol / L potassium dihydrogen phosphate buffer solution (pH = 3.5) - acetonitrile (55:45), and the detection wavelength was 208 nm. Accurately measure 10 μL each of the reference solution and the test solution, inject them into the liquid chromatograph, record the chromatograms, calculate the mass of cevelexta sodium in each device and filter paper using the external standard method, calculate the total delivery amount and delivery efficiency, and the results are shown in Table 6.

[0137] 2. Investigation of aerodynamic particle size distribution

[0138] Referring to Chapter 0951 of Part IV of the 2020 Chinese Pharmacopoeia, the aerodynamic particle size of cevelexostat sodium lyophilized powder was determined using a new generation pharmaceutical cascade impactor (NGI): 0.3 g of cevelexostat sodium lyophilized powder was weighed, reconstituted with 5 mL of water for injection, and nebulized using a PARI eflow nebulizer. The assembled impactor and L-shaped connecting tube were pre-cooled in a cooling device (5°C) for at least 90 minutes. Measurements were started within 5 minutes of removing it from the cooling device, and the airtightness was checked. The flow rate was set to 15 L / min for 3 minutes. After nebulization, the sample was thoroughly washed with methanol, and the active substances at each stage were collected. Three parallel tests were performed. Quantitative analysis was performed using HPLC under the same chromatographic conditions. Data were analyzed using Copley data analysis software version 3.10 to calculate the fine particle dose (FPD), fine particle fraction (FPF), and median mass aerodynamic diameter (MMAD). All samples were tested three times. The results are shown in Table 6, and the percentages of sediment at each level are shown in Table 6. Figure 2 .

[0139] Table 6 Results of Atomization Characteristics Evaluation

[0140]

[0141] As shown in Table 6, the lyophilized powders prepared using cevelexostat sodium nanocrystal suspension (Comparative Examples 2-3), cevelexostat sodium liposome suspension (Comparative Examples 4-5), and cevelexostat sodium phospholipid complex suspension (Examples 1-2) exhibited no significant differences in atomization characteristics such as delivery rate, total delivery volume, fine particle dosage, and median aerodynamic diameter. However, the lyophilized powder prepared using the cevelexostat sodium phospholipid complex suspension (Examples 1-2) showed a significantly higher percentage of particles smaller than 5 μm, with a fine particle fraction (FPF) exceeding 70%. Furthermore, from... Figure 2 It can be seen that the lyophilized powder prepared from the sodium nanocrystal suspension (Comparative Examples 2-3) has more particles deposited at level 2-3, the lyophilized powder prepared from the sodium liposome suspension (Comparative Examples 4-5) has more particles deposited at level 2-4, and the lyophilized powder prepared from the sodium phospholipid complex suspension (Examples 1-2) has more particles deposited at level 4-5. This indicates that the lyophilized powder prepared from the sodium phospholipid complex suspension of the present invention has better aerodynamic properties and is more suitable for penetrating deep into the alveoli and pulmonary microvessels for the treatment of ALI / ARDS.

[0142] Experimental Example 9: Stability Study of Cevelexta Sodium Lyophilized Powder

[0143] This experiment investigated the stability of lyophilized powders prepared from the sodium cevelestat suspensions of Examples 2, 3, and 5 (prepared using the same method as in Example 1) at 0, 1, 2, 3, and 6 months. The results are shown in Table 7.

[0144] Table 7. Stability results of lyophilized sodium cevelestatin powder

[0145]

[0146] As shown in Table 7, the lyophilized powders prepared using sodium cetylstatin nanocrystal suspension, sodium cetylstatin liposome suspension, and sodium cetylstatin phospholipid complex suspension all had impurity A <1.0% and total impurities <2% within 6 months, all exhibiting good stability.

[0147] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0148] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. A suspension of cipamidex sodium characterized in that, It is a mixed solution containing an osmotic pressure regulator and a surfactant, sodium cefixime and porous phospholipid particles, which are dispersed by shearing stirring and then homogenized to obtain; the mass ratio of sodium cefixime to porous phospholipid particles is 1.2-1.4:1; the osmotic pressure regulator is sodium chloride; and the surfactant is Tween 80; The pH of the mixed solution is 6.5-8.5; The porous phospholipid particles are prepared by the following method: DSPC and CaCl2 are added into 70-85℃ water for injection in a molar ratio of 1-3:1, a pore-forming agent is added while stirring, shear stirring, high-pressure homogenization, and then spray drying; the d90 particle size of the porous phospholipid particles is 2-4 μm; the pore size of the porous phospholipid particles is 0.2-1 μm; the density of the porous phospholipid particles is 0.4-0.5 g / cm 3 ; the pore-forming agent is perfluorooctyl bromide or perfluoropropane. 3 ​ The stirring speed of the shearing stirring is 10,000 rpm-12,000 rpm, and the stirring time of the shearing stirring is 10 min-15 min; The homogenization is high-pressure homogenization or microfluidization, the homogenization pressure of the microfluidization is 15,000 psi-20,000 psi, the speed of the microfluidization is 20 mm / s-70 mm / s, and the homogenization times of the microfluidization are 2-3 times.

2. The sodium cevimeline suspension according to claim 1, characterized in that, The pH of the mixed solution is 6.5-7.5; And / or, the mass-volume ratio of sodium cefixime to the mixed solution is 40 mg-60 mg:1 mL.

3. The sodium cevimeline suspension of claim 2, wherein, The pH of the mixed solution is 6.5-7.0, and the mass-volume ratio of sodium cefixime to the mixed solution is 50 mg-60 mg:1 mL.

4. The sodium cefixime suspension of claim 1, wherein, In the mixed solution, the concentration of sodium chloride is 8 mg / mL-10 mg / mL; and the concentration of Tween 80 is 0.01 mg / mL-10.0 mg / mL; And / or, the molar ratio of DSPC to CaCl2 is 1.5-2.5:1; And / or, the ratio of the mass of DSPC and CaCl2 to the volume of water for injection is 3-15 g:100 mL; And / or, the volume ratio of water for injection to porogen is 1-4:1; And / or, the speed of the shearing stirring is 8,000 rpm-10,000 rpm; And / or, the pressure of the high-pressure homogenization is 140 MPa-180 MPa; and the number of times of the high-pressure homogenization is 3-6 times; And / or, the process parameters of the spray drying include: inlet temperature 90°C-135°C; outlet temperature 68°C-72°C; feed speed 2 mL / min-8 mL / min; and total gas flow rate 425 L / min-525 L / min.

5. The sodium cevimeline suspension of claim 4, wherein, In the mixed solution, the concentration of sodium chloride is 8.5 mg / mL-9.5 mg / mL; the concentration of Tween 80 is 0.01 mg / mL-5.0 mg / mL; the molar ratio of DSPC to CaCl2 is 1.8-2.2:1; the ratio of the mass of DSPC and CaCl2 to the volume of water for injection is 4.5-5.5 g:100 mL; the porogen is perfluorooctyl bromide; the volume ratio of water for injection to porogen is 3-4:1; the pressure of the high-pressure homogenization is 160 MPa-170 MPa; and the number of times of the high-pressure homogenization is 4-5 times.

6. The sodium cevimeline suspension of claim 1, wherein, The stirring speed of the shearing stirring is 10000 rpm-11000 rpm; And / or, the stirring time of the shearing stirring is 10 min-13 min; And / or, the homogenization is microfluidization.

7. The sodium cevimeline suspension of claim 6, wherein, The stirring speed of the shearing stirring is 10000 rpm-10050 rpm; And / or, the stirring time of the shearing stirring is 10 min-11 min.

8. The sodium cevimeline suspension of claim 6, wherein, The homogenization pressure of the microfluidization is 19000 psi-20000 psi; And / or, the speed of the microfluidization is 40 mm / s-60 mm / s; And / or, the homogenization times of the microfluidization is 2 times.

9. A process for the preparation of the suspension of sodium cevimeline according to any one of claims 1 to 8, characterized in that, It comprises the following steps: adding the sodium sivelestat and porous phospholipid particles into a mixed solution containing an osmotic pressure regulator and a surfactant, dispersing by shearing stirring, and then homogenizing.

10. The method of preparing the suspension of cipemastat sodium according to claim 9, characterized in that, The stirring speed of the shearing stirring is 10000 rpm-12000 rpm; And / or, the stirring time of the shearing stirring is 10 min-15 min; And / or, the homogenization is high-pressure homogenization or microfluidization.

11. The method of preparing the sodium cevimeline suspension according to claim 10, characterized in that, The pressure of the high-pressure homogenization is 1000 bar-1400 bar, and the high-pressure homogenization time is 10 min-30 min; The homogenization pressure of the microfluidization is 15000 psi-20000 psi; and the speed of the microfluidization is 20 mm / s-70 mm / s; The homogenization times of the microfluidization is 2 times-3 times.

12. The method of preparing the sodium cevimeline suspension according to claim 11, characterized in that, The pressure of the high-pressure homogenization is 1100 bar-1300 bar, and the high-pressure homogenization time is 10 min-20 min; The homogenization pressure of the microfluidization is 18000 psi-20000 psi; the speed of the microfluidization is 40 mm / s-60 mm / s; and the homogenization times of the microfluidization is 2 times.

13. The method of preparing the sodium cevimeline suspension according to claim 12, characterized in that, The pressure of the high-pressure homogenization is 1150 bar-1250 bar, and the high-pressure homogenization time is 10 min-15 min; The homogenization pressure of the microfluidization is 19000 psi-20000 psi.

14. The method of preparing the sodium cevimeline suspension according to claim 9, characterized in that, The stirring speed of the shearing stirring is 10000 rpm-11000 rpm; And / or, the stirring time of the shearing stirring is 10 min-13 min.

15. The method of preparing the sodium cevimeline suspension according to claim 14, characterized in that, The stirring speed of the shearing stirring is 10000 rpm-10050 rpm; And / or, the stirring time of the shearing stirring is 10 min-11 min.

16. A sodium cevimeline lyophilized powder characterized in that, It is obtained by spray freeze-drying of the sodium sivelestat suspension according to any one of claims 1-8.

17. A sodium cipamfaste aerosol inhalant, characterized in that, It comprises the sodium sivelestat freeze-dried powder according to claim 16.

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