Inhalation formulation for treating IPF disease and preparation method therefor

Abstract

An inhalation formulation for treating an IPF disease. The inhalation formulation is an aerosol inhalation liquid formulation, an inhalation aerosol, a dry powder inhaler or a soft mist inhaler. The inhalation formulation contains an active drug, and the active drug is any one of apremilast, ibudilast, roflumilast, crisaborole, and difamilast. A preparation process for the inhalation formulation is introduced into a conventional formulation. An active component in a conventional oral formulation is prone to being destroyed in the digestive tract, and has low bioavailability, a liver first pass effect, and a slow onset of action. An inhalation formulation capable of significantly alleviating IPF is prepared. Tests prove that the prepared inhalation formulation can effectively increase the bioavailability, and improve the clinical use effect.

Description

An inhalation preparation for treating IPF disease and its preparation method Technical Field

[0001] The present invention relates to the technical field of inhalants, and in particular to an inhalation preparation for treating IPF disease and a preparation method thereof. Background Art

[0002] Idiopathic pulmonary fibrosis (IPF) is a non-neoplastic lung disease characterized by the formation of scar tissue in the lungs without any known trigger, followed by chronic, progressive fibrosis, and an unstoppable decline in lung function, progressive respiratory failure, and a high mortality rate. IPF is a rare disease that is usually progressive and causes severe disability, affecting approximately 50,000 people worldwide. Currently, the recommended treatments for IPF are pirfenidone and nintedanib, but these two drugs have limited efficacy in preventing disease progression and improving quality of life, and are also associated with tolerability issues.

[0003] Current IPF treatments are taken orally, while some compounds in development are administered subcutaneously or intravenously.

[0004] Despite extensive research, there are currently no inhaled formulations specifically for clinical use in the treatment of IPF. Summary of the Invention

[0005] In response to the problems existing in the above-mentioned prior art, the present invention provides an inhalation preparation for treating IPF disease and a preparation method thereof. The prepared inhalation preparation has a simple production process, is portable, has mild respiratory tract irritation, and has improved bioavailability.

[0006] The present invention is implemented through the following technical solutions:

[0007] An inhalation preparation for treating IPF disease, wherein the inhalation preparation is a nebulized inhalation liquid preparation, an inhalation aerosol, an inhalation powder inhalation or a soft mist;

[0008] The inhalation preparation contains an active drug, and the active drug is any one of apremilast, ibudilast, crisaborole, and defamister.

[0009] Furthermore, the active drug includes but is not limited to apremilast, ibudilast, crisaborole, and defamister.

[0010] Furthermore, the inhalation preparation is an inhalation liquid preparation.

[0011] Furthermore, the inhalation preparation is a nebulized inhalation liquid preparation.

[0012] Furthermore, the aerosol inhalation liquid preparation is composed of an active drug and a pharmaceutical excipient A;

[0013] The pharmaceutical excipient A comprises a solvent, a nonionic surfactant, a pH buffer, an osmotic pressure regulator and a complexing agent.

[0014] Furthermore, the solvent is deionized water;

[0015] Furthermore, the nonionic surfactant includes but is not limited to Tween, Span, polyol, and poloxamer;

[0016] The pH buffer includes but is not limited to acetic acid-sodium acetate, phosphoric acid-sodium phosphate, citric acid-sodium citrate, and its pH value is 4.0-6.0;

[0017] The osmotic pressure regulator includes but is not limited to sodium chloride, propylene glycol, glycerol, and glucose;

[0018] The complexing agent is EDTA or its salt.

[0019] Furthermore, the nonionic surfactant is any one of Tween, Span, polyol and poloxamer.

[0020] Furthermore, the method for preparing the aerosol inhalation liquid preparation comprises the following preparation steps:

[0021] Step S1. dissolving the osmotic pressure regulator and the complexing agent in a solvent, stirring until completely dissolved, adding the active drug, and stirring until completely dissolved to obtain solution A;

[0022] Step S2. adding the solvent to the nonionic surfactant and dispersing the mixture uniformly to obtain solution B;

[0023] Step S3. Add solution B to solution A, stir evenly, add the remaining solvent, and add pH buffer to adjust the pH.

[0024] Furthermore, the nebulizer inhalation agent is used in a nebulizer inhalation device, which includes but is not limited to Aerogen® (vibrating screen nebulization), Omron® (air compression nebulization), Yuyue® (ultrasonic nebulization), etc.

[0025] Furthermore, the inhalation preparation is an inhalation aerosol.

[0026] Furthermore, the inhalation aerosol is composed of an active pharmaceutical ingredient and a pharmaceutical excipient B;

[0027] The pharmaceutical excipient B comprises a hydrofluoroalkane propellant, a polyethylene glycol surfactant and a cosolvent.

[0028] Furthermore, the active pharmaceutical ingredient is prepared by micronization, wherein the micronization process includes but is not limited to air flow milling, spray drying, spray freeze drying, recrystallization, and ball milling.

[0029] Further, the hydrofluoroalkane propellant includes but is not limited to 1,1,1,2,3,3,3-heptafluoro-n-propane or 1,1,1,2-tetrafluoroethane;

[0030] Polyethylene glycol includes but is not limited to PEG200, PEG400, and PEG1000;

[0031] Co-solvents include, but are not limited to, propylene glycol or ethanol.

[0032] Further, the active drug comprises 1-3% w / w of the total formulation;

[0033] The polyethylene glycol surfactant accounts for 0.01-1% w / w of the total preparation.

[0034] Furthermore, the method for preparing the inhalation aerosol comprises adding the micronized active pharmaceutical ingredient into an aluminum can, weighing it, adding a surfactant and a co-solvent, quickly sealing the aluminum can with a metering valve, adding a propellant through the metering valve, and then ultrasonicating the filled aerosol can for 30 seconds to obtain the inhalation aerosol.

[0035] Furthermore, the inhalation aerosol is used for a device for inhaling aerosol, and the device for inhaling aerosol includes but is not limited to Ventolin®, Lixinping®, Jishu®, etc.

[0036] Furthermore, the inhalation preparation is an inhalation powder.

[0037] Furthermore, the inhalation powder consists of an active drug and a carrier;

[0038] The carrier includes but is not limited to lactose, mannose, chitosan, DPPC, and DSPC.

[0039] Furthermore, the content ratio of the active drug and the carrier is (5-100):(1-95).

[0040] Furthermore, the active drug particle size D90 is 1-10 μm;

[0041] The carrier particle size D90 is 3-200 μm.

[0042] Furthermore, the preparation method of the inhalation powder comprises the following steps:

[0043] S01. The active drug is micronized to obtain a first component;

[0044] S02. The first component is mixed with the carrier to obtain a second component;

[0045] S03. Prepare the second component into a dry powder preparation and load it into the corresponding device for standby use.

[0046] Furthermore, the micronization process includes but is not limited to air flow milling, spray drying, spray freeze drying, recrystallization, and ball milling;

[0047] Mixing methods include, but are not limited to, high shear mixing, air flow mixing, and three-dimensional mixing.

[0048] Furthermore, the device in S03 includes but is not limited to a Spiriva® device, a Seretide® device, a TurboSynbic® device or a Breezhaler® device.

[0049] Furthermore, the inhalation preparation is a soft mist.

[0050] Furthermore, the soft mist is composed of an active pharmaceutical ingredient and a pharmaceutical excipient C;

[0051] The pharmaceutical excipients C include solvents, additives and pH regulators.

[0052] Furthermore, the active pharmaceutical ingredient is prepared by micronization, wherein the micronization process includes but is not limited to air flow milling, spray drying, spray freeze drying, recrystallization, and ball milling.

[0053] Further, the solvent is deionized water;

[0054] The additive is EDTA or its salt;

[0055] The pH regulators are citric acid, hydrochloric acid and sodium hydroxide, and the pH range of the preparation is 2-6.

[0056] Further, the active drug concentration is 1ug / 100mL-1g / 100mL;

[0057] Furthermore, the preparation method of the soft mist is as follows: the additives and pH regulator are added to a beaker, part of the solvent is added to dissolve, the micronized active drug is added after dissolution, and the solvent is added to the required concentration, the pH is measured after stirring evenly, and the drug is put into a flexible plastic bottle for standby use.

[0058] Furthermore, the soft mist is administered through a soft mist device, which includes but is not limited to Spiriva Respimat and the like.

[0059] Beneficial effects of the present invention:

[0060] 1. This invention incorporates an inhalation preparation process into traditional formulations. The active ingredients in traditional oral formulations are easily destroyed in the digestive tract, resulting in low bioavailability, a first-pass effect in the liver, a slow onset of action, and significant systemic side effects. This invention produces an inhalation formulation that significantly improves IPF. Experimental studies have demonstrated that the prepared inhalation formulation can effectively increase bioavailability and enhance clinical efficacy.

[0061] 2. Currently, the only two preferred IPF treatments are nintedanib and pirfenidone. In this invention, we have demonstrated the effectiveness of PDE inhibitors in alleviating and improving IPF pulmonary fibrosis. Furthermore, we have modified the drug into an inhaled formulation, avoiding many of the drawbacks of oral formulations in the treatment of lung diseases, demonstrating significant innovation. BRIEF DESCRIPTION OF THE DRAWINGS

[0062] The accompanying drawings are used to further explain the present invention and constitute a part of the specification. Together with the embodiments of the present invention, they are used to explain the present invention and do not constitute a limitation of the present invention. In the accompanying drawings:

[0063] Figure 1 is a graph showing the body weight of mice in each group;

[0064] Figure 2 shows MASSON staining of the lungs of mice in each group;

[0065] Figure 3 shows HE staining of the lungs of mice in each group;

[0066] Figure 4 shows the detection of MDA and HYP content in the lungs of mice in each group;

[0067] Figures 5 and 6 are weight test diagrams of mice in each group in more examples and comparative examples;

[0068] FIG7 shows MASSON staining of the lungs of mice in more groups of Examples and Comparative Examples;

[0069] FIG8 shows HE staining of the lungs of mice in more groups of Examples and Comparative Examples;

[0070] FIG9 shows the detection of MDA and HYP content in the lungs of mice in more groups of Examples and Comparative Examples. DETAILED DESCRIPTION

[0071] The technical solution of the present invention is further described in detail below with reference to specific embodiments, but the protection scope of the present invention is not limited to the following embodiments.

[0072] IPF (idiopathic pulmonary fibrosis) is a distinct disease from chronic obstructive pulmonary disease (COPD), asthma, chronic bronchitis, and cystic fibrosis. IPF, short for idiopathic pulmonary fibrosis, is a chronic, progressive, fibrotic interstitial lung disease. This disease is characterized by the replacement of normal lung tissue with scar tissue (fibrosis), leading to a gradual hardening and thickening of the lung structure, which impairs gas exchange and makes breathing difficult. The exact cause of IPF is unknown, hence the term "idiopathic," but it may be linked to genetic factors, long-term smoking, certain viral infections, gastroesophageal reflux, environmental factors, and an abnormal immune system response.

[0073] The following are descriptions of several specific embodiments, comparative examples, and test examples of the present invention for treating IPF:

[0074] Example 1

[0075] The liquid preparation for aerosol inhalation of this embodiment includes an active pharmaceutical ingredient, a solvent, a nonionic surfactant, a pH buffer, an osmotic pressure regulator, and a complexing agent.

[0076] The active pharmaceutical ingredient is roflumilast, and the dosage is 20 μg / mL;

[0077] The solvent is water;

[0078] The nonionic surfactant is Tween 80, and the dosage is 100 mg / L;

[0079] The pH buffer is phosphate-sodium phosphate, and its pH value is 4.0;

[0080] The osmotic pressure regulator is sodium chloride, and the dosage is 9 mg / mL;

[0081] The complexing agent is EDTA-2Na, and the dosage is 200 mg / L.

[0082] The osmotic pressure regulator sodium chloride and the chelating agent EDTA-2Na are dissolved in 1 / 3 volume of solvent (water), stirred until dissolved, and then micronized roflumilast is added and stirred until dissolved; non-ionic surfactant Tween 80 is added to 1 / 3 volume of solvent (water), and after uniform dispersion, it is added to the aforementioned solution, stirred evenly, and then solvent (water) is added to the full amount, and buffer is added to adjust the pH to 5.0 to obtain the product.

[0083] The micronization process of the preparation process is air flow mixing;

[0084] The feed pressure of the air flow milling is 6 bar, and the discharge pressure of the air flow milling is 6 bar.

[0085] Example 2

[0086] The liquid preparation for aerosol inhalation of this embodiment includes an active pharmaceutical ingredient, a solvent, a nonionic surfactant, a pH buffer, an osmotic pressure regulator, a complexing agent, and the like.

[0087] The active pharmaceutical ingredient is Apremilast, and the dosage is 20 μg / mL;

[0088] The solvent is water;

[0089] The nonionic surfactant is Tween 80, and the dosage is 100 mg / L;

[0090] The pH buffer is phosphate-sodium phosphate, and its pH value is 4.0;

[0091] The osmotic pressure regulator is sodium chloride, and the dosage is 9 mg / mL;

[0092] The complexing agent is EDTA-2Na, and the dosage is 200 mg / L.

[0093] The preparation method of the apremilast aerosol inhalation liquid preparation is as follows:

[0094] The osmotic pressure regulator sodium chloride and the chelating agent EDTA-2Na are dissolved in 1 / 3 volume of solvent (water), stirred until dissolved, and then micronized apremilast is added and stirred until dissolved; non-ionic surfactant Tween 80 is added to 1 / 3 volume of solvent (water), and after uniform dispersion, the mixture is added to the aforementioned solution, stirred evenly, and then solvent (water) is added to the full amount, and buffer is added to adjust the pH to 5.0 to obtain the product.

[0095] The micronization process of the preparation process is air flow mixing;

[0096] The feed pressure of the air flow milling is 6 bar, and the discharge pressure of the air flow milling is 6 bar.

[0097] Example 3

[0098] The inhalation aerosol of this embodiment includes an active pharmaceutical ingredient, a hydrofluoroalkane propellant, a polyethylene glycol surfactant, and a cosolvent.

[0099] The active drug is crisaborole, which accounts for 2% w / w of the total preparation;

[0100] The propellant is 1,1,1,2-tetrafluoroethane, which accounts for 96% w / w of the total preparation;

[0101] The co-solvent is ethanol, which accounts for 1% w / w of the propellant.

[0102] The polyethylene glycol surfactant is PEG1000, accounting for 1% w / w of the total amount of the preparation.

[0103] The preparation method of the crisaborole inhalation aerosol is as follows:

[0104] The micronized active ingredient crisaborole is added into an aluminum can and weighed, and then the surfactant PEG1000 and the co-solvent ethanol are added. The aluminum can is quickly sealed with a metering valve, and the propellant 1,1,1,2-tetrafluoroethane is added through the metering valve. The filled aerosol can is then ultrasonicated for 30 seconds to obtain the product.

[0105] The micronization process of the preparation process is air flow mixing;

[0106] The feed pressure of the air flow milling is 6 bar, and the discharge pressure of the air flow milling is 6 bar.

[0107] Example 4

[0108] The inhalation aerosol of this embodiment includes an active pharmaceutical ingredient, a hydrofluoroalkane propellant, a polyethylene glycol surfactant, and a cosolvent.

[0109] The active pharmaceutical ingredient is Apremilast, which accounts for 2% w / w of the total preparation;

[0110] The propellant is 1,1,1,2-tetrafluoroethane, which accounts for 96% w / w of the total preparation;

[0111] The co-solvent is ethanol, which accounts for 1% w / w of the propellant.

[0112] The polyethylene glycol surfactant is PEG1000, accounting for 1% w / w of the total amount of the preparation.

[0113] The preparation method of the Apremilast inhalation aerosol is as follows:

[0114] The micronized active ingredient apremilast is added to an aluminum can and weighed. Then, the surfactant PEG1000 and the co-solvent ethanol are added. The aluminum can is quickly sealed with a metering valve. The propellant 1,1,1,2-tetrafluoroethane is then added through the metering valve. The filled aerosol can is then ultrasonicated for 30 seconds to obtain the product.

[0115] The micronization process of the preparation process is air flow mixing;

[0116] The feed pressure of the air flow milling is 6 bar, and the discharge pressure of the air flow milling is 6 bar.

[0117] Example 5

[0118] The dry powder inhalation preparation of this embodiment includes the following raw materials: active drug, carrier;

[0119] The active drug is roflumilast, which accounts for 20% of the total amount of the preparation;

[0120] The carrier is lactose monohydrate, which accounts for 80% of the total amount of the preparation;

[0121] The preparation method of the roflumilast dry powder inhalation preparation is as follows:

[0122] The active drug Roflumilast is subjected to air flow pulverization to obtain a first component, which is then mixed with the carrier lactose monohydrate to obtain a uniform mixture. After spheroidization, a spherical particle sample is obtained and loaded into a Dubao Xinbico device.

[0123] In the roflumilast dry powder inhalation preparation, the feed pressure of the active drug during airflow milling is 6 bar, the discharge pressure during airflow milling is 6 bar, the particle size is D90 4.4 μm, and the carrier particle size is D90 7.2 μm.

[0124] The mixing method is air flow mixing, wherein the feed pressure of the air flow milling is 1 bar, and the discharge pressure of the air flow milling is 6 bar.

[0125] Example 6

[0126] The dry powder inhalation preparation of this embodiment includes the following raw materials: active drug, carrier;

[0127] The active drug is Apremilast, which accounts for 20% of the total amount of the preparation;

[0128] The carrier is lactose monohydrate, which accounts for 80% of the total amount of the preparation;

[0129] The preparation method of the Apremilast dry powder inhalation preparation is as follows:

[0130] The active drug Apremilast is subjected to air flow pulverization to obtain the first component, which is then mixed with the carrier lactose monohydrate to obtain a uniform mixture. After spheroidization, a spherical particle sample is obtained and loaded into the Dubao Xinbico® device.

[0131] In the apremilast dry powder inhalation preparation, the feed pressure of the active drug during airflow milling is 6 bar, the discharge pressure during airflow milling is 6 bar, the particle size is D90 4.6 μm, and the carrier particle size is D90 7.5 μm.

[0132] The mixing method is air flow mixing, wherein the feed pressure of the air flow milling is 1 bar, and the discharge pressure of the air flow milling is 6 bar.

[0133] Example 7

[0134] The soft mist of this embodiment includes an active pharmaceutical ingredient, a solvent, an additive, and a pH adjuster.

[0135] The active pharmaceutical ingredient is Apremilast, and the dosage is 0.5 mg / mL;

[0136] The solvent is water;

[0137] The additive is EDTA, and the dosage is 0.1 mg / mL;

[0138] The pH regulator is citric acid, and the dosage is 0.03 mg / mL;

[0139] Add additives and citric acid to a beaker, add some solvent to dissolve, add micronized apremilast after dissolution, add water to 200 mL, stir evenly and measure the pH to 3.94, then put into a flexible plastic bottle for later use.

[0140] The micronization process of the preparation process is air flow mixing;

[0141] The feed pressure of the air flow milling is 6 bar, and the discharge pressure of the air flow milling is 6 bar.

[0142] Comparative Example 1

[0143] The difference between this comparative example and the embodiment is that the preparation used is oral apremilast.

[0144] Comparative Example 2

[0145] The difference between this comparative example and the embodiment is that the preparation used is oral nintedanib.

[0146] Test Example 1

[0147] According to the 2020 edition of the Pharmacopoeia of the People's Republic of China, General Rule 0951 of Part IV, the inhalation preparations prepared in Examples 1-6 were measured using Apparatus 3 according to the method for measuring the aerodynamic characteristics of fine particles of inhalation preparations. The measurement results are shown in Table 1.

[0148]

[0149] Table 1 Fine particle (FPF) measurement results of Examples 1-7

[0150] Test Example 2

[0151] Pharmacodynamic experiments

[0152] Pulmonary fibrosis model establishment: 6-8 week old C57BL6J mice were selected and adapted for one week; model group, apremilast inhalation (Example 2) treatment group, roflumilast inhalation (Example 5) treatment group, crisaborole inhalation (Example 3) treatment group, apremilast oral treatment group (Comparative Example 1) and nintedanib oral (Comparative Example 2, nintedanib is a commonly used drug for treating IPF in the prior art) treatment group was instilled with bleomycin solution (3 mg / kg / 50 μl) through the trachea on the first day, and the control group was treated with an equal amount of normal saline. Treatment started one week later for a period of 2 weeks, for a total of 3 weeks. Bleomycin solution was prepared with sterile PBS to the desired concentration of bleomycin dry powder (non-surgical method of bleomycin transtracheal injection into mouse lungs: mice were anesthetized with isoflurane and hung on a surgical board at a 70° angle. Bleomycin was instilled through the trachea using a 200 μl pipette).

[0153] The effects of apremilast inhalation treatment group, roflumilast inhalation treatment group, crisaborole inhalation treatment group, apremilast oral treatment group and nintedanib oral treatment group on lung injury in mice with pulmonary fibrosis were evaluated.

[0154] Body weight measurement: Weigh the mice at a fixed time every morning and record the weight, and observe the changes in the weight of mice in each group at different times.

[0155] HE staining and Masson staining: Mouse lung tissue was obtained and fixed in 4% paraformaldehyde for 24 hours, rinsed overnight in running water, dehydrated with standard alcohol, embedded in paraffin, and cut into 5 μm sections for HE and Masson staining, respectively. After clearing with xylene, the sections were mounted with neutral gum. Pathological changes in the lungs were observed under a light microscope and photographed.

[0156] Lung malondialdehyde (MDA) and hydroxyproline (HYP) content testing: Appropriate amounts of lung tissue from each group of mice were homogenized (grinded and centrifuged at 3000 rpm for 10 minutes, and the supernatant was collected for later use). MDA and HYP levels in the lungs were measured according to the kit instructions. MDA, representing malondialdehyde, is a product of lipid peroxidation and reflects oxidative damage, while HYP, representing hydroxyproline levels, reflects the degree of lung fibrosis.

[0157] Statistical Analysis: Graphpad Prism 9.0 software was used to analyze the experimental data. All data were tested for normality and presented as mean ± standard deviation (mean ± SD). One-way ANOVA was used to compare the means of multiple groups. P < 0.05 was considered statistically significant. The results are shown in Figures 1 to 4.

[0158] 1) PDEi treatment can effectively improve bleomycin-induced weight loss in mice, and the inhalation effect is better than oral administration

[0159] See Figure 1: Compared with the control group, the weight of mice was significantly reduced after bleomycin treatment for one week; compared with the model group, the weight of mice in the apremilast inhalation treatment group, roflumilast inhalation treatment group, crisaborole inhalation treatment group, and apremilast oral treatment group was significantly restored, and there was no significant difference in the weight recovery of mice in the nintedanib oral treatment group; compared with the apremilast oral treatment group, the weight recovery of mice in the apremilast inhalation treatment group, roflumilast inhalation treatment group, and crisaborole inhalation treatment group was more obvious.

[0160] 2) PDEi treatment reduces the pathological damage of pulmonary fibrosis and the inhalation effect is better than oral administration

[0161] As shown in Figure 2, MASSON staining showed that compared with the control group, the model group had obvious pathological changes, with thickening of the alveolar interstitium and a significant increase in collagen fiber deposition; compared with the model, the apremilast inhalation treatment group, the roflumilast inhalation treatment group, and the crisaborole inhalation treatment group had fewer pathological changes in the lungs of mice, with thin alveolar interstitium and clear boundaries of each part; compared with the inhalation group, the alveolar interstitium of mice in the oral apremilast and nintedanib groups was slightly thickened, with a few collagen fibers visible (see arrows in Figure 2). The arrows in the figure indicate the collagen deposition, as shown in Figure 2: there was no collagen fiber deposition in the lungs of the control group; the collagen fibers in the lungs of the model group increased significantly; the collagen fibers in the apremilast, roflumilast, and crisaborole inhalation groups were significantly reduced compared with the model group; the collagen deposition in the oral apremilast and nintedanib groups was reduced compared with the model group, but the degree of reduction was worse than that in the inhalation group.

[0162] As shown in Figure 3, HE staining showed that compared with the control group, the lung tissue structure of the mice in the model group was abnormal, with significant thickening of the alveolar wall, unclear alveolar structure, obvious fibrosis, and obvious inflammatory cell infiltration. Compared with the model, the abnormal lung tissue structure of mice in the apremilast inhalation treatment group, roflumilast inhalation treatment group, and crisaborole inhalation treatment group was improved, some alveoli were inflated, the alveolar wall was thinned, and very few inflammatory cells were visible; compared with the inhalation group, the abnormal alveolar structure of mice in the oral apremilast and nintedanib groups was improved, the alveolar wall was slightly thickened, some alveolar structures were unclear, and inflammatory cell infiltration was visible (see arrows in Figure 3). The arrows in the figure indicate the alveolar structure and inflammatory cell infiltration, as shown in Figure 3: the alveolar structure of the control group was clearly visible, without inflammatory cell infiltration; the lungs of the model group were dense, the alveolar cavity structure disappeared, and there was a large amount of inflammatory cell infiltration; the alveolar structure of the apremilast, roflumilast, and crisaborole inhalation groups was significantly improved compared with the model group, without obvious inflammatory cell infiltration; oral apremilast and nintedanib both had an improvement effect on the alveolar structure compared with the model group, with a small amount of inflammatory cell infiltration.

[0163] 3) Inhaled PDEi can effectively reduce lung MDA content and HYP levels

[0164] As shown in Figure 4, compared with the control group, the MDA and HYP levels in the lungs of the model group were significantly increased, and decreased significantly after inhalation treatment. Oral apremilast and nintedanib treatment showed a slight alleviation trend of MDA and HYP in the lungs, but there was no statistical significance. The results showed that apremilast inhalation treatment, roflumilast inhalation treatment, and crisaborole inhalation treatment can effectively reduce the oxidative stress level and collagen deposition of pulmonary fibrosis.

[0165] Nintedanib is a commonly used drug for the treatment of IPF in the prior art, but its limited efficacy and significant systemic side effects leave a significant gap in the selection of therapeutic agents for IPF. In summary, the present invention demonstrates that oral nintedanib, through a side-by-side comparison of the efficacy of IPF, demonstrates superior therapeutic effects compared to drugs such as apremilast, roflumilast, and crisaborole, demonstrating its innovative nature.

[0166] The following are other embodiments of the present invention and further description thereof:

[0167] Example 7

[0168] The soft mist of this embodiment includes an active pharmaceutical ingredient, a solvent, an additive, and a pH adjuster.

[0169] The active pharmaceutical ingredient is Apremilast, and the dosage is 0.5 mg / mL;

[0170] The solvent is water;

[0171] The additive is EDTA, and the dosage is 0.1 mg / mL;

[0172] The pH regulator is citric acid, and the dosage is 0.03 mg / mL;

[0173] Add additives and citric acid to a beaker, add some solvent to dissolve, add micronized apremilast after dissolution, add water to 200 mL, stir evenly and measure the pH to 3.94, then put into a flexible plastic bottle for later use.

[0174] The micronization process of the preparation process is air flow mixing;

[0175] The feed pressure of the air flow milling is 6 bar, and the discharge pressure of the air flow milling is 6 bar.

[0176] Example 8

[0177] The inhalation aerosol of this embodiment includes an active pharmaceutical ingredient, a hydrofluoroalkane propellant, a polyethylene glycol surfactant, and a cosolvent.

[0178] The active pharmaceutical ingredient is ibudilast, which accounts for 2% w / w of the total preparation;

[0179] The propellant is 1,1,1,2-tetrafluoroethane, which accounts for 96% w / w of the total preparation;

[0180] The co-solvent is ethanol, which accounts for 1% w / w of the propellant.

[0181] The polyethylene glycol surfactant is PEG1000, accounting for 1% w / w of the total amount of the preparation.

[0182] The preparation method of the ibudilast dry inhalation aerosol is as follows:

[0183] The micronized active ingredient ibudilast is added to an aluminum can and weighed. The surfactant PEG1000 and the co-solvent ethanol are then added. The aluminum can is quickly sealed with a metering valve. The propellant 1,1,1,2-tetrafluoroethane is then added through the metering valve. The filled aerosol can is then ultrasonicated for 30 seconds to obtain the product.

[0184] The micronization process of the preparation process is air flow mixing;

[0185] The feed pressure of the air flow milling is 6 bar, and the discharge pressure of the air flow milling is 6 bar.

[0186] Example 9

[0187] The dry powder inhalation preparation of this embodiment includes the following raw materials: active drug, carrier;

[0188] The active drug is defamilastat, which accounts for 20% of the total amount of the preparation;

[0189] The carrier is lactose monohydrate, which accounts for 80% of the total amount of the preparation;

[0190] The preparation method of the difamist dry powder inhalation preparation is as follows:

[0191] The active drug defamilastat is subjected to air flow pulverization to obtain a first component, which is then mixed with the carrier lactose monohydrate to obtain a uniform mixture. The mixture is then subjected to spheroidization to obtain a spherical particle sample, which is then loaded into a Dubao Xinbico device.

[0192] In the defamilastat dry powder inhalation preparation, the feed pressure of the active drug during airflow milling is 6 bar, the discharge pressure during airflow milling is 6 bar, the particle size D90 is 4.4 μm, and the carrier particle size D90 is 7.2 μm.

[0193] The mixing method is air flow mixing, wherein the feed pressure of the air flow milling is 1 bar, and the discharge pressure of the air flow milling is 6 bar.

[0194] Example 10

[0195] The dry powder inhalation preparation of this embodiment includes the following raw materials: active drug, carrier;

[0196] The active drug is ibudilast, which accounts for 20% of the total amount of the preparation;

[0197] The carrier is lactose monohydrate, which accounts for 80% of the total amount of the preparation;

[0198] The preparation method of the ibudilast dry powder inhalation preparation is as follows:

[0199] The active drug ibudilast is subjected to air flow pulverization to obtain the first component, which is then mixed with the carrier lactose monohydrate to obtain a uniform mixture. After spheroidization, a spherical particle sample is obtained and loaded into the Dubao Xinbico® device.

[0200] In the ibudilast dry powder inhalation preparation, the feed pressure of the active drug during airflow milling is 6 bar, the discharge pressure during airflow milling is 6 bar, the particle size D90 is 4.4 μm, and the carrier particle size D90 is 7.2 μm.

[0201] The mixing method is air flow mixing, wherein the feed pressure of the air flow milling is 1 bar, and the discharge pressure of the air flow milling is 6 bar.

[0202] According to the method of the above-mentioned test example 1, the test example 3 was carried out

[0203] According to the 2020 edition of the Pharmacopoeia of the People's Republic of China, General Rule 0951 of Part IV, the inhalation preparations prepared in Examples 8-10 were measured using Apparatus 3 according to the method for determining the aerodynamic characteristics of fine particles of inhalation preparations. The results are shown in Table 2.

[0204]

[0205] Table 2 Fine particle (FPF) measurement results of Examples 8-10

[0206] Comparative Examples 3 and 4 are added on the basis of the aforementioned Comparative Examples 1 and 2, as follows:

[0207] Comparative Example 3

[0208] The difference between this comparative example and the embodiment is that the preparation used is oral ibudilast.

[0209] Comparative Example 4

[0210] The difference between this comparative example and the embodiment is that the preparation used is oral defamilastat.

[0211] Furthermore, based on the above-mentioned Experimental Example 2, Experimental Example 4 was carried out.

[0212] Pharmacodynamic experiments

[0213] Pulmonary fibrosis model establishment: 6-8 week-old C57 / BL6J mice were selected and acclimated for one week. The mice were divided into a blank control group, a model group, a premilast inhalation treatment group (Example 2), a roflumilast inhalation treatment group (Example 5), a crisaborole inhalation treatment group (Example 3), an ibudilast inhalation treatment group (Example 10), a defamilast inhalation treatment group (Example 9), an ibudilast oral treatment group (Comparative Example 3), a defamilast oral treatment group (Comparative Example 4), and a nintedanib oral treatment group (Comparative Example 2; nintedanib is a commonly used drug for the treatment of IPF in the prior art). The n = 8 mice per group. On the first day, mice in the model group and each treatment group were instilled with a bleomycin solution (3 mg / kg / 50 μl) via tracheal instillation. The control group was treated with an equal amount of normal saline. One week later, treatment began in each treatment group for two weeks, for a total of three weeks. Bleomycin solution was prepared by preparing bleomycin powder with sterile PBS to the desired concentration (non-surgical method of transtracheal infusion of bleomycin into mouse lungs: mice were anesthetized by isoflurane inhalation and suspended on a surgical board at a 70° angle, and bleomycin was instilled through the trachea using a 200 μl pipette).

[0214] The effects of apremilast inhalation treatment group, roflumilast inhalation treatment group, crisaborole inhalation treatment group, ibudilast inhalation treatment group, defamilast inhalation treatment group, apremilast oral treatment group, ibudilast oral treatment group, defamilast oral treatment group and nintedanib oral treatment group on lung injury in mice with pulmonary fibrosis were evaluated.

[0215] Body weight measurement: Weigh the mice at a fixed time every morning and record the weight, and observe the changes in the weight of mice in each group at different times.

[0216] HE staining and Masson staining: Mouse lung tissue was obtained and fixed in 4% paraformaldehyde for 24 hours, rinsed overnight in running water, dehydrated with standard alcohol, embedded in paraffin, and serially sectioned (5 μm) for HE and Masson staining, respectively. After clearing with xylene, the sections were mounted with neutral gum. Observed and photographed under a light microscope.

[0217] Detection of malondialdehyde (MDA) and hydroxyproline (HYP) content in the lungs: An appropriate amount of lung tissue from each group of mice was homogenized (grinded and centrifuged at 3000 rpm for 10 minutes, and the supernatant was collected for later use). The MDA and HYP levels in the lungs were detected according to the instructions of the kit (MDA is malondialdehyde, a product of lipid peroxidation, reflecting the degree of oxidative damage; HYP (hydroxyproline) levels reflect the degree of pulmonary fiber deposition).

[0218] Statistical Analysis: Graphpad Prism 9.0 software was used to analyze the experimental data. All data were tested for normality and presented as mean ± standard deviation (mean ± SD). One-way ANOVA was used to compare the means of multiple groups. P < 0.05 was considered statistically significant. The results are shown in Figures 5 to 9.

[0219] 1) PDEi treatment can effectively improve bleomycin-induced weight loss in mice, and the inhalation effect is better than oral administration

[0220] See Figures 5 and 6: Compared with the control group, the weight of mice was significantly reduced after bleomycin treatment for one week; compared with the model group, the weight of mice in the apremilast inhalation treatment group, roflumilast inhalation treatment group, criborole inhalation treatment group, ibudilast inhalation treatment group, defamilast inhalation treatment group, apremilast oral treatment group, ibudilast oral treatment group, and defamilast oral treatment group was significantly restored, and there was no significant difference in the weight recovery of mice in the nintedanib oral treatment group compared with the model group; compared with the oral treatment group, the weight recovery of mice in each drug inhalation treatment group was more obvious. As can be seen from Figures 5 and 6, compared with the 7th day after the end of modeling, the weight of the mice in the model group did not recover significantly on the 14th day, while the weight of the mice in the apremilast inhalation treatment group, roflumilast inhalation treatment group, criborole inhalation treatment group, ibudilast inhalation treatment group, defamilast inhalation treatment group, apremilast oral treatment group, ibudilast oral treatment group, and defamilast oral treatment group recovered significantly, while the weight of the mice in the nintedanib oral treatment group did not recover significantly.

[0221] 2) PDEi treatment reduces the pathological damage of pulmonary fibrosis and the inhalation effect is better than oral administration

[0222] As shown in Figure 7, MASSON staining showed that compared with the control group, the model group had obvious pathological changes, with thickening of alveolar interstitial tissue, significantly increased collagen fiber deposition, and disappearance of normal alveolar ventilation structure. Compared with the model, the lung damage of mice in the apremilast inhalation treatment group, roflumilast inhalation treatment group, crisaborole inhalation treatment group, ibudilast inhalation treatment group, and defamilast inhalation treatment group was significantly milder, with no thickening of alveolar interstitial tissue and clear alveolar structure, which was similar to that of the control group. Compared with the inhalation group, the apremilast oral treatment group, ibudilast oral treatment group, and defamilast oral treatment group had significantly milder lung damage. The alveolar interstitium of mice in the oral treatment group and the nintedanib oral treatment group was slightly thickened, and a small amount of collagen fibers were visible (see the arrows in Figure 7). The arrows in the figure indicate the collagen deposition. As shown in Figure 7: there was no collagen fiber deposition in the lungs of the control group; the collagen fibers in the lungs of the model group increased significantly; the collagen fibers in the apremilast, roflumilast, crisaborole, ibudilast, and defamilast inhalation groups were significantly reduced compared with the model group; the collagen deposition of apremilast, ibudilast, defamilast, and nintedanib oral administration was reduced compared with the model group, but the degree of reduction was less than that of the inhalation group.

[0223] As shown in Figure 8, HE staining showed that compared with the control group, the lung tissue structure of the mice in the model group was abnormal, with significant thickening of the alveolar wall, unclear alveolar structure, obvious fibrosis, and obvious inflammatory cell infiltration. Compared with the model, the lung tissue structure of the mice in the apremilast inhalation treatment group, the roflumilast inhalation treatment group, the crisborole inhalation treatment group, the ibudilast inhalation treatment group, and the defamilast inhalation treatment group was significantly improved, with some alveoli inflated, no significant thickening of the alveolar wall, and less inflammatory cell infiltration; compared with the inhalation treatment group, the alveolar structural abnormalities of the mice in the apremilast oral treatment group, the ibudilast oral treatment group, the defamilast oral treatment group, and the nintedanib oral treatment group were improved, with slightly thickened alveolar wall, unclear alveolar structure, and visible inflammatory cell infiltration (see Figure 8). 8), the arrows in the figure indicate the alveolar structure and inflammatory cell infiltration, as shown in Figure 8: the alveolar structure of the control group was clearly visible, without inflammatory cell infiltration; the lungs of the model group were dense, the alveolar cavity structure disappeared, and there was a large amount of inflammatory cell infiltration; the alveolar structure of the inhalation treatment groups of apremilast, roflumilast, crisaborole, ibudilast, and defamilast was significantly improved compared with the model group, without obvious inflammatory cell infiltration; apremilast, ibudilast, defamilast and nintedanib oral administration all had an improvement effect on the alveolar structure compared with the model group, with a small amount of inflammatory cell infiltration.

[0224] 3) Inhaled PDEi can effectively reduce lung MDA content and HYP levels

[0225] As shown in Figure 9, compared with the control group, the MDA and HYP levels in the lungs of the model group were significantly increased, and significantly decreased after inhalation treatment. The MDA and HYP levels in the lungs of mice in the oral apremilast, ibudilast, defamilast and nintedanib treatment groups showed a slight downward trend but no statistical significance. The results showed that apremilast inhalation treatment, roflumilast inhalation treatment, crisaborole inhalation treatment, ibudilast inhalation treatment, and defamilast inhalation treatment can effectively reduce the oxidative stress level and collagen deposition of pulmonary fibrosis.

[0226] As mentioned above, nintedanib is a commonly used drug for treating IPF in the prior art, but because of its general efficacy and the large systemic side effects, there is a large gap in the selection of therapeutic drugs for IPF. The present invention compares the efficacy of oral nintedanib in treating IPF in parallel, confirming that drugs such as apremilast, roflumilast, and crisaborole can show better therapeutic effects and are creative. That is, in the present invention, the active drug is any one of apremilast, ibudilast, roflumilast, crisaborole, and defamilast; and the active drug includes but is not limited to apremilast, ibudilast, roflumilast, crisaborole, and defamilast.

[0227] In summary, the inhalation preparation for treating IPF disease and the preparation method thereof provided by the present invention have a simple production process, are portable, and can effectively improve the bioavailability of the active ingredient.

[0228] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above and that the invention can be embodied in other specific forms without departing from the spirit or essential characteristics of the invention. Therefore, the embodiments should be considered in all respects as illustrative and non-restrictive, and the scope of the invention is defined by the appended claims rather than the foregoing description, and it is intended that all variations that come within the meaning and range of equivalents of the claims be embraced therein.

[0229] In addition, it should be understood that although this specification is described in terms of implementation methods, not every implementation method contains only one independent technical solution. This narrative method of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment can also be appropriately combined to form other implementation methods that can be understood by those skilled in the art.

Claims

1. An inhalation preparation for treating IPF disease, characterized in that: The inhalation preparation is an inhalation liquid preparation, an inhalation aerosol, an inhalation powder spray or a soft mist; The inhalation preparation contains an active drug, and the active drug is any one of apremilast, ibudilast, crisaborole, and defamister.

2. The inhalation preparation according to claim 1, characterized in that The inhalation preparation is an inhalation liquid preparation.

3. The inhalation preparation according to claim 2, characterized in that The atomized inhalation liquid preparation is composed of an active drug and a pharmaceutical excipient A; The pharmaceutical excipient A comprises a solvent, a nonionic surfactant, a pH buffer, an osmotic pressure regulator and a complexing agent.

4. The inhalation preparation according to claim 3, characterized in that The solvent is deionized water; The nonionic surfactant is any one of Tween, Span, polyol and poloxamer; The pH buffer is any one of acetic acid-sodium acetate, phosphoric acid-sodium phosphate, and citric acid-sodium citrate, and its pH value is 4.0-6.0; The osmotic pressure regulator is any one of sodium chloride, propylene glycol, glycerol, and glucose; The complexing agent is EDTA or its salt.

5. The method for preparing the inhalation preparation according to any one of claims 2 to 4, characterized in that: The method comprises the following preparation steps: Step S1. dissolving the osmotic pressure regulator and the complexing agent in a solvent, stirring until completely dissolved, adding the active drug, stirring until completely dissolved, to obtain solution A; Step S2. adding the solvent to the nonionic surfactant and dispersing the mixture evenly to obtain a solution B; Step S3. Add solution B to solution A, stir evenly, add the remaining solvent, and add pH buffer to adjust the pH to obtain an inhalation aerosol.

6. The inhalation preparation according to claim 1, characterized in that The inhalation preparation is an inhalation aerosol.

7. The inhalation preparation according to claim 6, characterized in that The inhalation aerosol is composed of an active pharmaceutical ingredient and a pharmaceutical excipient B; The pharmaceutical excipient B comprises a hydrofluoroalkane propellant, a polyethylene glycol surfactant and a cosolvent.

8. The inhalation preparation according to claim 7, characterized in that The active pharmaceutical ingredient is prepared by micronization, wherein the micronization is any one of air flow pulverization, spray drying, spray freeze drying, recrystallization, and ball milling.

9. The inhalation preparation according to claim 7, characterized in that The hydrofluoroalkane propellant is 1,1,1,2,3,3,3-heptafluoro-n-propane or 1,1,1,2-tetrafluoroethane; The polyethylene glycol is any one of PEG200, PEG400 and PEG1000; The co-solvent is propylene glycol or ethanol.

10. The method for preparing the inhalation preparation according to any one of claims 6 to 9, characterized in that: The micronized active pharmaceutical ingredient is added into an aluminum can, weighed, and then a surfactant and a co-solvent are added. The aluminum can is quickly sealed with a metering valve, and then the propellant is added through the metering valve. The filled aerosol can is then ultrasonicated for 30 seconds to obtain an inhalation aerosol.

11. The inhalation preparation according to claim 1, characterized in that The inhalation preparation is an inhalation powder.

12. The inhalation preparation according to claim 11, characterized in that The inhalation powder consists of an active drug and a carrier; The carrier is at least one of lactose, mannose, chitosan, DPPC and DSPC.

13. The inhalation preparation according to claim 12, characterized in that The content ratio of the active drug and the carrier is (5-100):(1-95).

14. The inhalation preparation according to claim 12, characterized in that The particle size D90 of the active drug is 1-10 μm; The carrier particle size D90 is 3-200 μm.

15. The method for preparing the inhalation preparation according to any one of claims 11 to 14, characterized in that: The following steps are involved: S01. The active drug is micronized to obtain a first component; S02. The first component is mixed with the carrier to obtain a second component; S03. Prepare the second component into a dry powder preparation and load it into a corresponding device for standby use.

16. The method for preparing the inhalation preparation according to claim 15, characterized in that: Micronization is any one of air flow pulverization, spray drying, spray freeze drying, recrystallization, and ball milling; The mixing method is any one of high shear mixing, air flow mixing and three-dimensional mixing.

17. The inhalation preparation according to claim 1, characterized in that The inhalation preparation is a soft mist.

18. The inhalation preparation according to claim 17, characterized in that The soft mist is composed of active drugs and pharmaceutical excipients C; The pharmaceutical excipient C includes a solvent, an additive and a pH adjuster.

19. The inhalation preparation according to claim 18, characterized in that The solvent is deionized water; The additive is EDTA or its salt; The pH regulator is citric acid, hydrochloric acid, and sodium hydroxide, and the pH range of the preparation is 2-6.

20. The method for preparing the inhalation preparation according to any one of claims 17 to 19, characterized in that: Add additives and pH adjusters to a beaker, add some solvent to dissolve, add micronized active drug after dissolution, and add solvent to the required concentration, stir evenly and measure the pH, put into a flexible plastic bottle and set aside.

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