Fudosteine pharmaceutical composition, preparation method therefor, and use thereof

Abstract

Disclosed is a pharmaceutical composition containing Fudosteine, comprising the following components: a pharmaceutically active ingredient, a pH adjuster, and water. The concentration of Fudosteine in the composition ranges from 20 mg / mL to 200 mg / mL, and the pH value is between 3.0 and 6.0. The pharmaceutically active ingredient is one or more of Fudosteine, a pharmaceutically acceptable salt thereof, and a hydrate thereof. The pharmaceutical composition containing Fudosteine does not comprise an antioxidant, a surfactant, and a metal chelating agent, and can be prepared into an atomized inhalation preparation.

Description

Fudostertan pharmaceutical compositions, their preparation methods and applications

[0001] This application claims priority to an earlier application filed on November 11, 2024, with the China National Intellectual Property Administration, patent application number 2024115967995, entitled "Fodostan Pharmaceutical Composition, Preparation Method Thereof, and Application". The entire contents of the earlier application are incorporated herein by reference. Technical Field

[0002] This invention relates to a folic acid pharmaceutical composition, its preparation method, and its application. Background Technology

[0003] Fudosteine ​​is a novel respiratory mucolytic that effectively inhibits the excessive formation of goblet cells that secrete viscous mucus in the respiratory tract, thereby inhibiting the production of highly viscous fucoidin, reducing the viscosity of sputum, and making it easier to cough up. In addition, fudosteine ​​can enhance the secretory function of serous trachea, thus inhibiting tracheal inflammation.

[0004] Fudosteine, chemically named (-)-(R)-2-amino-3-(3-hydroxypropylthio)propionic acid, is a novel and potent expectorant with six effects: expectoration, expectoration, expectoration thinning, expectoration, anti-inflammation, and anti-oxidation. It was first launched in Japan in 2001 and is a cysteine ​​derivative with a steine-based skeleton developed by Mitsubishi Pharmaceutical Co., Ltd. and SS Pharmaceutical Co., Ltd. It has multiple pharmacological effects on chronic respiratory diseases: it inhibits the proliferation of respiratory epithelial cells, normalizes the trehalose / sialic acid ratio in sputum, restores the state of ciliary transport of airway secretions, and has anti-inflammatory effects; it has the advantages of strong efficacy, few side effects, wide range of indications, and great market potential; it is suitable for expectoration of chronic respiratory diseases such as bronchial asthma, chronic bronchitis, bronchiectasis, pulmonary tuberculosis, pneumoconiosis, emphysema, atypical mycobacterial infection, and diffuse bronchiolitis.

[0005] Currently marketed fodosteine-related drugs are all oral dosage forms (including oral solutions, tablets, capsules, and granules). Common adverse reactions include rash, nausea, indigestion and itching, decreased sensation, abdominal discomfort, and diarrhea, because fodosteine ​​damages the gastric mucosal barrier. To reduce the adverse reactions of fodosteine, existing technology has improved it into a nebulized inhalation solution formulation (see Chinese Patent Publication No. CN109925300A), allowing the drug to be administered locally through the lungs, effectively avoiding damage to the gastric mucosal barrier. Due to the manufacturing process of fodosteine, trace amounts of metal ions often exist in its solution. The presence of metal ions catalyzes oxidation reactions, leading to the oxidative decomposition of fodosteine, and also causes safety risks and side effects. To address these issues, CN109925300A adds EDTA as a metal ion chelating agent (antioxidant), thereby reducing the metal ion content in the product formulation and improving its stability. However, EDTA has a certain degree of mucosal irritation, and there is a risk of inducing adverse reactions such as coughing and asthma when inhaling EDTA-containing nebulized inhalation solutions. Chinese patent documents CN117442587A and CN115068450B disclose a fudostein nebulized inhalation solution composition, which does not contain metal ion chelating agents, but has a narrow pH range requirement, strict controllability in production, and is prone to causing coughing during inhalation.

[0006] Therefore, finding a solution to reduce the content of free metal ions while simultaneously reducing the irritation and production controllability caused by metal ion chelating agents has become an urgent technical problem to be solved. Summary of the Invention

[0007] The technical problem solved by this invention is to overcome the high free metal ion content, significant side effects, and poor production feasibility of existing fodostein nebulized inhalation solution compositions. This invention provides a fodostein pharmaceutical composition, its preparation method, and its applications. The fodostein composition of this invention exhibits good stability, low irritation, a small effective dose, high efficacy, good patient compliance, and strong production controllability. Furthermore, it does not contain antioxidants and / or surfactants, resulting in a lower incidence of adverse reactions, better safety, and promising market prospects.

[0008] This invention provides a fodosteine ​​pharmaceutical composition comprising the following components: an active pharmaceutical ingredient, a pH adjuster, and water; wherein the concentration of fodosteine ​​in the composition is 20 mg / ml to 200 mg / ml, and the pH value is 3.0 to 6.0; wherein the active pharmaceutical ingredient is one or more of fodosteine, its pharmaceutically acceptable salts, and its hydrates.

[0009] According to an embodiment of the present invention, the fodosteine ​​pharmaceutical composition is a nebulized inhalation solution.

[0010] According to an embodiment of the present invention, the pH adjuster may be selected from inorganic bases, inorganic acids, buffer systems formed by inorganic acids and inorganic acid salts, organic acids, or buffer systems formed by organic acids and organic acid salts.

[0011] According to an embodiment of the present invention, the inorganic acid is selected from one or more of hydrochloric acid, sulfuric acid, and phosphoric acid; for example, hydrochloric acid.

[0012] According to an embodiment of the present invention, the inorganic acid salt is selected from one or more of sodium carbonate, sodium bicarbonate, disodium hydrogen phosphate, and sodium dihydrogen phosphate.

[0013] According to an embodiment of the present invention, the organic acid is selected from one or more of tartaric acid, lactic acid, citric acid, glacial acetic acid, and malic acid.

[0014] According to an embodiment of the present invention, the organic acid salt is selected from sodium citrate.

[0015] According to an embodiment of the present invention, the inorganic base is selected from one or more of sodium hydroxide, potassium hydroxide, sodium bicarbonate, and triethylamine.

[0016] According to embodiments of the present invention, the pH adjuster is further preferably selected from one or more of hydrochloric acid, sulfuric acid, tartaric acid, and sodium hydroxide. In some embodiments, the pH adjuster is hydrochloric acid or sodium hydroxide.

[0017] According to an embodiment of the present invention, the active pharmaceutical ingredient is preferably fodosteine.

[0018] According to an embodiment of the present invention, the pH value of the fudostein pharmaceutical composition can be 3.0-3.4, 3.4-4.0 or 4.1-6.0, for example 3.0, 3.4, 3.5, 3.7, 3.8, 4.0, 4.1, 4.5, 5.0, 5.5 or 6.0.

[0019] According to an embodiment of the present invention, when the pH adjuster is selected from inorganic acids or buffer systems formed by inorganic acids and inorganic acid salts (e.g., the pH adjuster is hydrochloric acid), the pH value of the fudosteine ​​pharmaceutical composition is 3.0 to 5.0, for example 3.0 to 4.0, 3.7 to 3.8 or 4.0 to 5.0, exemplary values ​​being 3.0, 3.4, 3.5, 3.7, 3.8, 4.0, 4.1, 4.5 or 5.0.

[0020] According to an embodiment of the present invention, when the pH adjuster is selected from inorganic bases (e.g., sodium hydroxide), the pH value of the fodosteine ​​pharmaceutical composition is greater than 5.0 and does not exceed 6.0, for example, 5.1–6.0 or 5.5–6.0, exemplarily 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, or 6.0. This composition has a near-neutral pH, which helps reduce the risk of coughing during inhalation.

[0021] According to an embodiment of the present invention, the concentration of the active pharmaceutical ingredient is preferably 50 mg / ml to 150 mg / ml, for example 50 mg / ml, 60 mg / ml, 70 mg / ml, 80 mg / ml, 90 mg / ml, 100 mg / ml, 110 mg / ml, 120 mg / ml, 130 mg / ml, 140 mg / ml or 150 mg / ml, wherein the concentration refers to the ratio of the mass of the active pharmaceutical ingredient to the volume of the fodosteine ​​pharmaceutical composition.

[0022] According to an embodiment of the present invention, the fudostein pharmaceutical composition does not contain one or more of antioxidants, surfactants and metal chelators, preferably does not contain antioxidants, surfactants and metal chelators.

[0023] According to an embodiment of the present invention, the surfactant is selected from one or more of Tween, Span, stearic acid, sodium dodecylbenzenesulfonate and lecithin.

[0024] According to an embodiment of the present invention, the antioxidant is selected from one or more of tert-butylhydroquinone, vitamin C (ascorbic acid), and vitamin E.

[0025] According to an embodiment of the present invention, the metal ion chelating agent is selected from ethylenediaminetetraacetic acid (EDTA).

[0026] According to an embodiment of the present invention, the water may be water for injection.

[0027] According to an embodiment of the present invention, the fodosteine ​​pharmaceutical composition comprises the following components: fodosteine, a pH adjuster and water, wherein the concentration of fodosteine ​​is 20 mg / ml to 200 mg / ml, and the pH adjuster is selected from inorganic acids or buffer systems formed by inorganic acids and inorganic acid salts (e.g., the pH adjuster is hydrochloric acid).

[0028] The pH value of the fodosteine ​​pharmaceutical composition is 3.0–5.0;

[0029] The fodosteine ​​pharmaceutical composition does not contain antioxidants, surfactants, or metal chelators.

[0030] According to an embodiment of the present invention, the fodosteine ​​pharmaceutical composition comprises the following components: fodosteine, a pH adjuster and water, wherein the concentration of fodosteine ​​is 20 mg / ml to 200 mg / ml, and the pH adjuster is selected from inorganic bases (e.g., sodium hydroxide).

[0031] The pH value of the fodosteine ​​pharmaceutical composition is greater than 5.0 and does not exceed 6.0;

[0032] The fodosteine ​​pharmaceutical composition does not contain antioxidants, surfactants, or metal chelators.

[0033] According to embodiments of the present invention, the fodosteine ​​pharmaceutical composition may be any of the following formulations:

[0034] Prescription 1: Fudosteine ​​100mg / ml, hydrochloric acid and water for injection, adjust pH to 3.7-3.8;

[0035] Prescription 2: Fudosteine ​​150 mg / ml, hydrochloric acid and water for injection, adjust pH to 3.7-3.8;

[0036] Prescription 3: Fudosteine ​​200 mg / ml, hydrochloric acid and water for injection, adjust pH to 3.7-3.8;

[0037] Prescription 4: Fudosteine ​​80 mg / ml, hydrochloric acid and water for injection, adjust pH to 3.7-3.8;

[0038] Prescription 5: Fudosteine ​​80 mg / ml, hydrochloric acid and water for injection, adjust pH to 3.7-3.8;

[0039] Prescription 6: Fudosteine ​​80 mg / ml, hydrochloric acid and water for injection, adjust pH to 3.0;

[0040] Prescription 7: Fudosteine ​​80 mg / ml, hydrochloric acid and water for injection, adjust pH to 3.5;

[0041] Prescription 8: Fudosteine ​​80 mg / ml, hydrochloric acid and water for injection, adjust pH to 4.0;

[0042] Prescription 9: Fudosteine ​​80 mg / ml, hydrochloric acid and water for injection, adjust pH to 4.5;

[0043] Prescription 10: Fudosteine ​​80 mg / ml, hydrochloric acid and water for injection, adjust pH to 5.0;

[0044] Prescription 11: Fudosteine ​​80 mg / ml, sodium hydroxide and water for injection, adjust pH to 5.5;

[0045] Prescription 12: Fudosteine ​​80 mg / ml, sodium hydroxide and water for injection, adjust pH to 6.0.

[0046] The present invention also provides a method for preparing the fudostein pharmaceutical composition, comprising mixing the components in the fudostein pharmaceutical composition.

[0047] According to an embodiment of the present invention, the preparation method further includes subjecting the mixed components to sterile filtration. The sterile filtration is preferably performed using a polyethersulfone membrane.

[0048] According to an embodiment of the present invention, the preparation method includes further packaging the fodosteine ​​pharmaceutical composition into a container. The container may be an ampoule or a vial.

[0049] According to an embodiment of the present invention, the material of the container is selected from any one of glass, polypropylene plastic, polyethylene plastic and polyester; preferably a glass ampoule or a low-density polyethylene ampoule.

[0050] According to an embodiment of the present invention, the container can be protected by nitrogen gas.

[0051] According to an embodiment of the present invention, the pharmaceutical composition is preferably packaged in a container, protected by nitrogen, and then filled and sealed. According to an embodiment of the present invention, the dissolved oxygen after filling and sealing is ≤1.0 mg / L.

[0052] The present invention also provides the use of the aforementioned fodosteine ​​pharmaceutical composition in the preparation of pharmaceutical formulations.

[0053] According to an embodiment of the present invention, the pharmaceutical preparation can be a nebulized inhalation preparation.

[0054] According to an embodiment of the present invention, the pharmaceutical preparation (e.g., a nebulized inhalation preparation) is a single dose, with a specification of 1.0 ml to 10.0 ml, preferably 1.0 ml to 5.0 ml, and more preferably 1.0 ml to 2.5 ml.

[0055] According to embodiments of the present invention, the pharmaceutical preparation is used to treat and / or prevent bronchial asthma, chronic wheezing bronchitis, bronchiectasis, pulmonary tuberculosis, pneumoconiosis, chronic obstructive emphysema, atypical mycobacterial disease, pneumonia, and / or diffuse bronchitis.

[0056] According to an embodiment of the present invention, the administration site of the fodosteine ​​composition is one or more of the following: nose, pharynx, trachea, esophagus and main bronchus, preferably the pharynx, trachea and main bronchus.

[0057] The present invention also provides the use of the aforementioned fudostein pharmaceutical composition in the preparation of medicaments for the treatment and / or prevention of bronchial asthma, chronic wheezing bronchitis, bronchiectasis, pulmonary tuberculosis, pneumoconiosis, chronic obstructive emphysema, atypical mycobacterial disease, pneumonia, and diffuse bronchitis.

[0058] The present invention also provides a method for treating and / or preventing diseases such as bronchial asthma, chronic wheezing bronchitis, bronchiectasis, pulmonary tuberculosis, pneumoconiosis, chronic obstructive emphysema, atypical mycobacterial disease, pneumonia and / or diffuse bronchitis, which provides a therapeutically effective amount of the fudostein pharmaceutical composition to patients in need.

[0059] The fodosteine ​​drug composition described herein can be used with a variety of nebulizers with different mechanisms, including but not limited to compressor nebulizers and screen nebulizers.

[0060] The atomizer is selected from one or more of the following: pari, Yuwell, Wududu, Yinghua Rongtai, and Medisana.

[0061] For example, the atomizer parameters include: flow rate of 5-30 L / min, such as 5 L / min, 10 L / min, 15 L / min, 25 L / min, 30 L / min, etc.; atomization time of 30-120 s, such as 30 s, 60 s, 90 s, 120 s, etc.; pre-cooling time of 60-180 min, such as 60 min, 90 min, 120 min, etc.; and pre-cooling temperature of 2-8℃, such as 5℃, etc.

[0062] For example, the aerodynamic particle size distribution results of the fudostein drug composition atomization are as follows: the percentage of fine particles (FPF%) is not less than 40% or is between 40% and 70%, such as 40.284%, 45.137%, 53.614%, 56.239%, 60.247%, and 65.483%; the amount of fine particles (FPD) is not less than 10 mg or is between 10 mg and 25 mg, such as 10.9401 mg and 12 mg. 0.4440mg, 13.8614mg, 15.1511mg, 17.2672mg, 20.8751mg, etc.; the mass median aerodynamic size distribution (MMAD) is not greater than 10μm or is 1μm~10μm, 3μm~6μm, such as 5μm, 4.9μm, 4.8μm, 4.6μm, 4.4μm, 4.3μm, 3.9μm, 3.8μm, 3.6μm, etc.

[0063] For example, the delivery rate is not less than 5 mg / min or is between 5 mg / min and 12 mg / min, such as 7.03 mg / min, 7.93 mg / min, 8.99 mg / min, 10.82 mg / min, etc.; the total delivery amount is not less than 20 mg or is between 20 and 110 mg or 20 and 70 mg, such as 24.38 mg, 25.89 mg, 33.77 mg, 60.17 mg, 99.39 mg; the residual amount is not more than 150 mg or is between 90 and 150 mg, such as 100.41 mg, 110.62 mg, 111.73 mg, 125.90 mg, 129.00 mg, 140.79 mg.

[0064] The present invention also provides a nebulizing assembly, including a nebulizing inhalation device (e.g., a nebulizer) and a fodosteine ​​pharmaceutical composition;

[0065] The nebulized inhalation device (e.g., a nebulizer) and the fodosteine ​​pharmaceutical composition have the limitations shown above.

[0066] Without violating common sense in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain various preferred embodiments of the present invention.

[0067] The reagents and raw materials used in this invention are all commercially available.

[0068] In this invention, room temperature refers to an ambient temperature of 10℃ to 35℃, preferably 25℃.

[0069] The beneficial effects of this invention are as follows: The fodosteine ​​nebulized inhalation formulation of this invention allows fodosteine ​​to act directly on the lungs, increasing the concentration of the drug administered to the respiratory tract or lungs. It has a rapid onset of action, avoids the first-pass effect of the liver and the degradation and destruction of the gastrointestinal tract, reducing damage to the patient's organs, maintaining efficacy, and improving bioavailability. Furthermore, the inhalation formulation has advantages such as minimal irritation, ease of use, good patient compliance, low toxicity, and suitability for long-term treatment, making it suitable for patients with dysphagia. The fodosteine ​​composition of this invention exhibits good stability (high-temperature stability, light stability, accelerated stability, and / or long-term stability), low irritation, a small effective dose, high efficacy, good patient compliance, and strong production controllability. It also does not contain antioxidants and / or surfactants and / or metal chelators, resulting in a lower incidence of adverse reactions, better safety, and good market prospects.

[0070] Terminology Definitions and Explanations

[0071] The term "multiple" refers to two or more physical mixtures, such as two, three or more physical mixtures.

[0072] The term "patient" refers to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, or primates, with humans being the most preferred.

[0073] The term "therapeutic effective amount" refers to the amount of an active compound or drug that researchers, veterinarians, physicians, or other clinicians are looking for in tissues, systems, animals, individuals, or humans to elicit a biological or medical response.

[0074] The term "comprising" includes a pharmaceutical composition consisting of the components listed above, and when the pharmaceutical composition consists of the components listed above, it does not exclude the presence (within specified limits) of impurities and / or isomers in the pharmaceutical composition. Detailed Implementation

[0075] The present invention is further illustrated below by way of embodiments, but the invention is not limited to the scope of the embodiments described herein. Experimental methods in the following embodiments that do not specify specific conditions were performed according to conventional methods and conditions, or as selected according to the product instructions.

[0076] Example 1

[0077] Prescription composition:

[0078] Fudostertan (100mg / ml) 10.0g

[0079] Add appropriate amount of hydrochloric acid to adjust the pH to 3.7–3.8.

[0080] Add water for injection to a final volume of 100 mL.

[0081] Preparation process:

[0082] 1) Purge the room temperature water for injection at 80% of the prescription volume with nitrogen for protection until the filling is completed, and control the dissolved oxygen to ≤1.0 mg / L;

[0083] 2) Add fodostatin and stir at room temperature until completely dissolved;

[0084] 3) Add 0.5 mol / L hydrochloric acid solution to adjust the pH to 3.7–3.8;

[0085] 4) Add water for injection to the prescribed amount and stir until well mixed;

[0086] 5) Filter the drug solution obtained in step 4) through a 0.22μm polyethersulfone filter membrane, fill each clean borosilicate glass ampoule with 2.0ml of drug solution, fill with nitrogen gas, and seal.

[0087] Example 2

[0088] Prescription composition:

[0089] Fudostertan (150mg / ml) 15.0g

[0090] Add appropriate amount of hydrochloric acid to adjust the pH to 3.7–3.8.

[0091] Add water for injection to a final volume of 100 mL.

[0092] Preparation process:

[0093] 1) Purge the room temperature water for injection at 80% of the prescription volume with nitrogen for protection until the filling is completed, and control the dissolved oxygen to ≤1.0 mg / L;

[0094] 2) Add fodostatin and stir at room temperature until completely dissolved;

[0095] 3) Add 0.5 mol / L hydrochloric acid solution to adjust the pH to 3.7–3.8;

[0096] 4) Add water for injection to the prescribed amount and stir until well mixed;

[0097] 5) Filter the drug solution obtained in step 4) through a 0.22μm polyethersulfone filter membrane, fill each clean borosilicate glass ampoule with 2.0ml of drug solution, fill with nitrogen gas, and seal.

[0098] Example 3

[0099] Prescription composition:

[0100] Fudosteine ​​(200mg / ml) 20.0g

[0101] Add appropriate amount of hydrochloric acid to adjust the pH to 3.7–3.8.

[0102] Add water for injection to a final volume of 100 mL.

[0103] Preparation process:

[0104] 1) Purge the room temperature water for injection at 80% of the prescription volume with nitrogen for protection until the filling is completed, and control the dissolved oxygen to ≤1.0 mg / L;

[0105] 2) Add fodostatin and stir at room temperature until completely dissolved;

[0106] 3) Add 0.5 mol / L hydrochloric acid solution to adjust the pH to 3.7–3.8;

[0107] 4) Add water for injection to the prescribed amount and stir until well mixed;

[0108] 5) Filter the drug solution obtained in step 4) through a 0.22μm polyethersulfone filter membrane, fill each clean borosilicate glass ampoule with 2.0ml of drug solution, fill with nitrogen gas, and seal.

[0109] Example 4

[0110] Prescription composition:

[0111] Fudostertan (80mg / ml) 24.0g

[0112] Add appropriate amount of hydrochloric acid to adjust the pH to 3.7–3.8.

[0113] Add water for injection to a final volume of 300 mL.

[0114] Preparation process:

[0115] 1) Add fodosteine ​​raw material to 80% of the prescribed amount of room temperature water for injection and stir at room temperature until completely dissolved;

[0116] 2) Add 0.5 mol / L hydrochloric acid solution to adjust the pH to 3.7–3.8;

[0117] 3) Add water for injection to the prescribed amount and stir until well mixed;

[0118] 4) Filter the drug solution obtained in step 3) through a 0.22μm polyethersulfone filter membrane, fill each clean borosilicate glass ampoule with 2.5ml of drug solution, and seal it.

[0119] Example 5

[0120] Prescription composition:

[0121] Fudostertan (80mg / ml) 16.0g

[0122] Add appropriate amount of hydrochloric acid to adjust the pH to 3.7–3.8.

[0123] Add water for injection to a final volume of 200 mL.

[0124] Preparation process:

[0125] 1) Purge the room temperature water for injection at 80% of the prescription volume with nitrogen for protection until the filling is completed, and control the dissolved oxygen to ≤1.0 mg / L;

[0126] 2) Add fodostatin and stir at room temperature until completely dissolved;

[0127] 3) Add 0.5 mol / L hydrochloric acid solution to adjust the pH to 3.7–3.8;

[0128] 4) Add water for injection to the prescribed amount and stir until well mixed;

[0129] 5) Filter the drug solution obtained in step 4) through a 0.22μm polyethersulfone filter membrane, fill 2.5ml of the drug solution into a low-density polyethylene ampoule, and seal it to obtain the final product.

[0130] Example 6

[0131] Prescription composition:

[0132] Fudostertan (80mg / ml) 16.0g

[0133] Add hydrochloric acid to a pH of 3.0.

[0134] Add water for injection to a final volume of 200 mL.

[0135] Preparation process:

[0136] 1) Purge the room temperature water for injection at 80% of the prescription volume with nitrogen for protection until the filling is completed, and control the dissolved oxygen to ≤1.0 mg / L;

[0137] 2) Add fodostatin and stir at room temperature until completely dissolved;

[0138] 3) Add 0.5 mol / L hydrochloric acid solution to adjust the pH to 3.0;

[0139] 4) Add water for injection to the prescribed amount and stir until well mixed;

[0140] 5) Filter the drug solution obtained in step 4) through a 0.22μm polyethersulfone filter membrane, fill each clean borosilicate glass ampoule with 2.5ml of drug solution, fill with nitrogen gas, and seal to obtain the final product.

[0141] Example 7

[0142] Prescription composition:

[0143] Fudostertan (80mg / ml) 16.0g

[0144] Add hydrochloric acid to a pH of 3.5.

[0145] Add water for injection to a final volume of 200 mL.

[0146] Preparation process:

[0147] 1) Purge the room temperature water for injection at 80% of the prescription volume with nitrogen for protection until the filling is completed, and control the dissolved oxygen to ≤1.0 mg / L;

[0148] 2) Add fodostatin and stir at room temperature until completely dissolved;

[0149] 3) Add 0.5 mol / L hydrochloric acid solution to adjust the pH to 3.5;

[0150] 4) Add water for injection to the prescribed amount and stir until well mixed;

[0151] 5) Filter the drug solution obtained in step 4) through a 0.22μm polyethersulfone filter membrane, fill each clean borosilicate glass ampoule with 2.5ml of drug solution, fill with nitrogen gas, and seal to obtain the final product.

[0152] Example 8

[0153] Prescription composition:

[0154] Fudostertan (80mg / ml) 16.0g

[0155] Add hydrochloric acid to adjust the pH to 4.0.

[0156] Add water for injection to a final volume of 200 mL.

[0157] Preparation process:

[0158] 1) Purge the room temperature water for injection at 80% of the prescription volume with nitrogen for protection until the filling is completed, and control the dissolved oxygen to ≤1.0 mg / L;

[0159] 2) Add fodostatin and stir at room temperature until completely dissolved;

[0160] 3) Add 0.5 mol / L hydrochloric acid solution to adjust the pH to 4.0;

[0161] 4) Add water for injection to the prescribed amount and stir until well mixed;

[0162] 5) Filter the drug solution obtained in step 4) through a 0.22μm polyethersulfone filter membrane, fill each clean borosilicate glass ampoule with 2.5ml of drug solution, fill with nitrogen gas, and seal to obtain the final product.

[0163] Example 9

[0164] Prescription composition:

[0165] Fudostertan (80mg / ml) 16.0g

[0166] Add hydrochloric acid to adjust the pH to 4.5.

[0167] Add water for injection to a final volume of 200 mL.

[0168] Preparation process:

[0169] 1) Purge the room temperature water for injection at 80% of the prescription volume with nitrogen for protection until the filling is completed, and control the dissolved oxygen to ≤1.0 mg / L;

[0170] 2) Add fodostatin and stir at room temperature until completely dissolved;

[0171] 3) Add 0.5 mol / L hydrochloric acid solution to adjust the pH to 4.5;

[0172] 4) Add water for injection to the prescribed amount and stir until well mixed;

[0173] 5) Filter the drug solution obtained in step 4) through a 0.22μm polyethersulfone filter membrane, fill each clean borosilicate glass ampoule with 2.5ml of drug solution, fill with nitrogen gas, and seal to obtain the final product.

[0174] Example 10

[0175] Prescription composition:

[0176] Fudostertan (80mg / ml) 16.0g

[0177] Add hydrochloric acid to a pH of 5.0.

[0178] Add water for injection to a final volume of 200 mL.

[0179] Preparation process:

[0180] 1) Purge the room temperature water for injection at 80% of the prescription volume with nitrogen for protection until the filling is completed, and control the dissolved oxygen to ≤1.0 mg / L;

[0181] 2) Add fodostatin and stir at room temperature until completely dissolved;

[0182] 3) Add 0.5 mol / L hydrochloric acid solution to adjust the pH to 5.0;

[0183] 4) Add water for injection to the prescribed amount and stir until well mixed;

[0184] 5) Filter the drug solution obtained in step 4) through a 0.22μm polyethersulfone filter membrane, fill each clean borosilicate glass ampoule with 2.5ml of drug solution, fill with nitrogen gas, and seal to obtain the final product.

[0185] Example 11

[0186] Prescription composition:

[0187] Fudostertan (80mg / ml) 16.0g

[0188] Add an appropriate amount of sodium hydroxide to bring the pH to 5.5.

[0189] Add water for injection to a final volume of 200 mL.

[0190] Preparation process:

[0191] 1) Purge the room temperature water for injection at 80% of the prescription volume with nitrogen for protection until the filling is completed, and control the dissolved oxygen to ≤1.0 mg / L;

[0192] 2) Add fodostatin and stir at room temperature until completely dissolved;

[0193] 3) Add 0.5 mol / L sodium hydroxide solution to adjust the pH to 5.5;

[0194] 4) Add water for injection to the prescribed amount and stir until well mixed;

[0195] 5) Filter the drug solution obtained in step 4) through a 0.22μm polyethersulfone filter membrane, fill each clean borosilicate glass ampoule with 2.5ml of drug solution, fill with nitrogen gas, and seal to obtain the final product.

[0196] Example 12

[0197] Prescription composition:

[0198] Fudostertan (80mg / ml) 16.0g

[0199] Add an appropriate amount of sodium hydroxide to bring the pH to 6.0.

[0200] Add water for injection to a final volume of 200 mL.

[0201] Preparation process:

[0202] 1) Purge the room temperature water for injection at 80% of the prescription volume with nitrogen for protection until the filling is completed, and control the dissolved oxygen to ≤1.0 mg / L;

[0203] 2) Add fodostatin and stir at room temperature until completely dissolved;

[0204] 3) Add 0.5 mol / L sodium hydroxide solution to adjust the pH to 6.0;

[0205] 4) Add water for injection to the prescribed amount and stir until well mixed;

[0206] 5) Filter the drug solution obtained in step 4) through a 0.22μm polyethersulfone filter membrane, fill each clean borosilicate glass ampoule with 2.5ml of drug solution, fill with nitrogen gas, and seal to obtain the final product.

[0207] Example 13

[0208] Prescription composition:

[0209] Fudostertan (80mg / ml) 16.0g

[0210] Add appropriate amount of hydrochloric acid to adjust the pH to 3.7–3.8.

[0211] Add water for injection to a final volume of 200 mL.

[0212] Preparation process:

[0213] 1) Purge the room temperature water for injection at 80% of the prescription volume with nitrogen for protection until the filling is completed, and control the dissolved oxygen to ≤1.0 mg / L;

[0214] 2) Add fodostatin and stir at room temperature until completely dissolved;

[0215] 3) Add 0.5 mol / L hydrochloric acid solution to adjust the pH to 3.7–3.8;

[0216] 4) Add water for injection to the prescribed amount and stir until well mixed;

[0217] 5) Filter the drug solution obtained in step 4) through a 0.22μm polyethersulfone filter membrane, fill each clean borosilicate glass ampoule with 2.5ml of drug solution, fill with nitrogen gas, and seal to obtain the final product.

[0218] Example 14 High Temperature Test

[0219] The samples from Examples 1 to 13 of this invention were placed at a high temperature (50°C) for 10 days and 30 days, respectively. On the 10th and 30th days, samples were taken to measure the appearance, pH value and related substances, and compared with the data of the same batch of samples on day 0. The results are shown in Tables 2, 3, 4 and 5 (in Tables 2-5, " / " means not detected and "-" means not detected).

[0220] The methods for detecting related substances are as follows:

[0221] 1. Related substances I: Determined by high performance liquid chromatography (Chinese Pharmacopoeia 2025 Edition, Part IV, General Chapter 0512). Operate in the dark.

[0222] Solvent: 0.9 g / L sodium octane sulfonate buffer solution - acetonitrile (94:6).

[0223] For the test solution, take 10 vials of this product, place the contents in the same container, accurately measure 1 ml, place it in a 20 ml volumetric flask, dilute to the mark with solvent, and shake well.

[0224] For the impurity I reference standard stock solution, accurately weigh approximately 4 mg of impurity I reference standard, place it in a 20 ml volumetric flask, dissolve and dilute with water to the mark, and shake well.

[0225] For the impurity II reference standard stock solution, accurately weigh approximately 4 mg of impurity II reference standard, place it in a 20 ml volumetric flask, add an appropriate amount of 1 mol / L hydrochloric acid solution to dissolve it, dilute with water to the mark, and shake well.

[0226] For the reference solution, accurately weigh approximately 4 mg of fodosteine ​​reference standard, place it in a 100 ml volumetric flask, add an appropriate amount of solvent, sonicate to dissolve, and dilute to the mark with solvent, then shake well. Accurately measure 2 ml of the above solution, place it in a 10 ml volumetric flask, dilute to the mark with solvent, and shake well to obtain the final solution.

[0227] For system suitability solution preparation, take appropriate amounts of fodosteine ​​inhalation solution, impurity I reference stock solution, and impurity II reference stock solution, dissolve them in solvent, and quantitatively dilute to prepare a mixed solution containing approximately 4.0 mg of fodosteine ​​and approximately 8 μg each of impurity I and impurity II per ml.

[0228] Chromatographic conditions: Octadecylsilane-bonded silica gel was used as the stationary phase (Waters XBridge C18, 4.6 mm × 250 mm, 5 μm or equivalent column); the mobile phase was 0.9 g / L sodium octanesulfonate buffer (0.9 g sodium octanesulfonate dissolved in 1000 ml of water, pH adjusted to 1.50 ± 0.05 with phosphoric acid) - acetonitrile (94:6); the flow rate was 1.0 ml / min; the column temperature was 35 °C; the detection wavelength was 210 nm; the injection volume was 20 μl; and the run time was 60 minutes.

[0229] System suitability requirements: In the chromatogram of the solution, the peaks of impurity I (double peak), fudostein, and impurity II should elute sequentially, and the resolution between each peak and its adjacent peaks should meet the requirements.

[0230] For the assay, accurately measure the test solution and the reference solution, inject them separately into the liquid chromatograph, and record the chromatograms.

[0231] If impurity peaks are present in the chromatogram of the test solution, they should be calculated based on peak area using the external standard method with correction factors. Impurity I (calculated as the sum of the two peak areas) (correction factor 0.76) and Impurity II (correction factor 0.85) should not exceed 0.2% of the labeled amount. Other individual impurities should not exceed 0.2% of the labeled amount. Except for Impurity I and Impurity II, the sum of the peak areas of all other impurities should not exceed 0.6% of the labeled amount. Peaks in the chromatogram of the test solution that are less than 0.02% of the labeled amount are negligible.

[0232] Note: The number of injection needles for blank solution must not be less than 5.

[0233] 2. Related substances II were determined according to high performance liquid chromatography (Chinese Pharmacopoeia 2025 Edition, Part IV, General Chapter 0512). The procedure was performed in the dark.

[0234] Solvent: water.

[0235] For the test solution, take 10 vials of this product, place the contents in the same container, accurately measure 1 ml, place it in a 20 ml volumetric flask, dilute to the mark with solvent, and shake well; accurately measure 5 ml of the above solution, place it in a 20 ml volumetric flask, dilute to the mark with solvent, and shake well to obtain the test solution.

[0236] For the impurity III reference standard stock solution, accurately weigh approximately 4 mg of impurity III reference standard, place it in a 20 ml volumetric flask, dissolve and dilute to the mark with 0.1 mol / L hydrochloric acid solution, and shake well.

[0237] For the impurity VII reference standard stock solution, accurately weigh approximately 4 mg of impurity VII reference standard, place it in a 20 ml volumetric flask, dissolve and dilute to the mark with 0.1 mol / L hydrochloric acid solution, and shake well.

[0238] For the reference solution, accurately weigh approximately 4 mg of fodosteine ​​reference standard, place it in a 100 ml volumetric flask, add an appropriate amount of solvent, sonicate to dissolve, and dilute to the mark with solvent, then shake well. Accurately measure 1 ml of the above solution, place it in a 20 ml volumetric flask, dilute to the mark with solvent, and shake well to obtain the reference solution.

[0239] System suitability solution (1) Take appropriate amounts of the stock solution of impurity I and impurity II reference standards under related substances I, and dilute them with solvent to prepare a solution containing approximately 2 μg of impurity I and impurity II per 1 ml.

[0240] System suitability solution (2) Take appropriate amounts of fodostein inhalation solution, impurity III reference stock solution and impurity VII reference stock solution, dissolve them in solvent and dilute quantitatively to prepare a mixed solution containing approximately 1.0 mg of fodostein and approximately 2 μg each of impurity III and impurity VII per 1 ml.

[0241] Chromatographic conditions: Octadecylsilane-bonded silica gel was used as the stationary phase (CAPCELL PAK C18 ACR S5, 4.6 mm × 250 mm, 5 μm or equivalent column); mobile phase A was 6.8 g / L potassium dihydrogen phosphate buffer solution (6.8 g of potassium dihydrogen phosphate dissolved in 1000 ml of water, pH adjusted to 4.0 with phosphoric acid), and mobile phase B was acetonitrile-water (80:20). Gradient elution was performed according to the table below; flow rate was 0.5 ml / min; column temperature was 30 °C; detection wavelength was 210 nm; injection volume was 20 μl.

[0242] Table 1 Gradient elution program

[0243] System suitability requirements: In the chromatogram of the system suitability solution (2), the peaks of impurity III, fudostein and impurity VII emerge in sequence, and the resolution between each peak and the adjacent peak should meet the requirements; the resolution between impurity VII and the adjacent peak should not be less than 1.0.

[0244] For the assay, accurately measure the test solution and the reference solution, inject them separately into the liquid chromatograph, and record the chromatograms.

[0245] If any impurity peaks are present in the chromatogram of the limit test solution, all chromatographic peaks in the system suitability solution (1) shall be deducted. Based on the peak area calculated using the principal component external standard method with correction factors, impurity III (correction factor 0.78) shall not exceed 0.2% of the labeled amount. Similarly, impurity VII (correction factor 1.5) shall not exceed 0.2% of the labeled amount, and other individual impurities shall not exceed 0.2% of the labeled amount. The sum of the peak areas of all impurities shall not exceed 1.0% of the labeled amount. Peaks in the chromatogram of the test solution that are less than 0.02% of the labeled amount shall be ignored.

[0246] 3. The dextrorotatory isomer was determined by high performance liquid chromatography (Chinese Pharmacopoeia 2025 Edition, Part IV, General Chapter 0512). The procedure was performed in the dark.

[0247] Solvent: water.

[0248] For the test solution, take 10 vials of this product, place the contents in the same container, accurately measure 2 ml, place it in a 20 ml volumetric flask, dilute to the mark with solvent, and shake well.

[0249] For the reference solution, accurately weigh approximately 4 mg of fodosteine ​​reference standard, place it in a 50 ml volumetric flask, add an appropriate amount of solvent, sonicate to dissolve, dilute to the mark with solvent, and shake well.

[0250] For system suitability solution, accurately weigh approximately 4 mg of the dextrorotatory isomer reference standard, place it in a 50 ml volumetric flask, add an appropriate amount of solvent, and sonicate to dissolve. Accurately measure 5 ml of fodosteine ​​inhalation solution, place it in the same volumetric flask, dilute to the mark with solvent, and shake well.

[0251] Chromatographic conditions: Octadecylsilane-bonded silica gel was used as the stationary phase (YMC Triart C18, 4.6 mm × 250 mm, 5 μm or equivalent column); chiral solution (1.32 g of D-phenylalanine and 1.0 g of copper sulfate dissolved in 1000 ml of water) - methanol (98:2) was used as the mobile phase; the flow rate was 0.6 ml per minute; the column temperature was 30 °C; the detection wavelength was 270 nm; the injection volume was 20 μl; and the run time was 30 minutes.

[0252] The system suitability requirements are that in the chromatogram of the reference solution, the fodosteine ​​peak and the dextrorotatory isomer peak should emerge sequentially, and the resolution between the two peaks should meet the requirements; the theoretical plate number, calculated based on the fodosteine ​​peak, should not be less than 2000.

[0253] For the assay, accurately measure the test solution and the reference solution, inject them separately into the liquid chromatograph, and record the chromatograms.

[0254] If a peak with the same retention time as the dextrorotatory isomer is present in the chromatogram of the test solution, the peak area should be calculated using the external standard method for the main component and should not exceed 1.0% of the labeled amount. Peaks in the chromatogram of the test solution that are less than 0.05% of the labeled amount should be ignored. Note: Inject the sample after equilibration under the above chromatographic conditions for at least 4 hours.

[0255] Table 2. Screening results of fodostatin concentration

[0256] The results of the high-temperature test showed that after 30 days of storage at 50°C, there were no significant differences in the properties, pH value and related substances I of samples with different concentrations. However, as the concentration of fodostein increased, the total impurities and isomers of related substances II showed an increasing trend (see Table 2).

[0257] Table 3 Results of Nitrogen Filling Process Study

[0258] The results of the high-temperature test showed that samples with the same concentration of nitrogen-filled and non-nitrogen-filled nitrogen showed no significant differences in properties, pH value, related substances I, related substances II, and isomers after being placed at 50°C for 30 days (see Table 3).

[0259] Table 4. Results of the study on packaging container materials

[0260] The results of the high-temperature test showed that the samples packaged in glass ampoules and polyethylene plastic ampoules did not differ significantly in properties, pH value, and related substances I and II after being placed at 50°C for 30 days (see Table 4).

[0261] Table 5 Results of pH screening test

[0262] The results of the pH 3.0 to pH 6.0 test showed that after being placed at a high temperature of 50℃ for 30 days, there were no significant differences in pH value, properties, and related substances I and II among samples with different pH values ​​(see Table 5).

[0263] Example 15

[0264] The stability of the sample from Example 11 was investigated under accelerated conditions (40±2℃, 75%±5%RH). The specific results are detailed in Table 6.

[0265] Table 6 Summary of Accelerated Stability Data in Example 11

[0266] Note: (1) Related substances I: The content of impurity I is the result of the sum of the peak areas of the two peaks; total impurities are the sum of all other impurity peaks except for impurity I and impurity II; the neglect limit is 0.02%.

[0267] (2) Related substances II: The neglect limit is 0.02%, and all known impurities are included in the total impurities.

[0268] The results in the table above show that after 6 months of acceleration, the product's stability is good and does not exceed the limit requirements.

[0269] Example 17 Pharmacokinetic Study of Sprague-Dawley Rats After Single Inhalation / Intravenous Administration

[0270] In Example 11, Sprague-Dawley rats were administered fodosteine ​​tablets (0.2 g / tablet, batch number: B241002J5D, Sichuan Kelun Pharmaceutical Co., Ltd.) via single inhalation and intravenous administration, respectively. The pharmacokinetic characteristics of fodosteine ​​in SD rats, as well as the relative and absolute bioavailability of the inhaled administration, were investigated. A total of 30 SD rats (half male and half female) were used in the pharmacokinetic study, divided into 5 groups of 6 rats each, all administered via single dose. The administration regimens are shown in Table 7.

[0271] Table 7 Dosing Regimen

[0272] The pharmacokinetic parameters of different groups are shown in Table 8:

[0273] Table 8 Summary of pharmacokinetic parameters for different administration methods

[0274] Note: F% represents absolute bioavailability, and F1% represents relative bioavailability.

[0275] The AUC of 13.2 mg / kg, 39.5 mg / kg and 118.2 mg / kg fodosteine ​​inhalation solution administered by a single inhalation was compared with that of intravenous administration. lastIn comparison, the absolute bioavailability (F%) of fodosteine ​​in male SD rats was 26.69%, 34.09%, and 64.71%, respectively, while the absolute bioavailability (F%) in female SD rats was 53.18%, 63.94%, and 78.16%, respectively; compared with the AUC of gavage administration... last In comparison, the relative bioavailability (F%) of fodosteine ​​in male SD rats was 26.15%, 102.28%, and 63.41%, respectively, while the relative bioavailability (F%) in female SD rats was 46.65%, 191.81%, and 68.57%, respectively.

[0276] Example 18 Active systemic anaphylaxis test administered to guinea pigs by inhalation

[0277] Thirty-six male Hartley guinea pigs were randomly divided into four groups of nine animals each. The negative control group was sensitized by inhalation of 0.9% sodium chloride injection for 30 minutes. The positive control group was sensitized by inhalation of 20 mg / mL chicken ovalbumin for 5 minutes. In Example 11, the low- and high-dose groups were sensitized by inhalation of the same drug for 10 and 30 minutes, respectively, with actual delivered doses of approximately 11 and 32 mg / kg. The challenge duration for each group was twice that of the initial sensitization. Sensitization was performed on days 1, 3, and 5. Fourteen days after the last sensitization (day 19), the first three animals in each group were challenged. Aerosol particle size distribution analysis was performed after administration on days 1, 5, and 19. The results are shown in Table 9.

[0278] Table 9. Actual average sensitization and aerosol particle size distribution parameters in Example 11

[0279] Under the experimental conditions, Hartley guinea pigs were sensitized by inhalation with fudostein inhalation solution of Example 11 at average delivery doses of 12.6 mg / kg and 37.7 mg / kg on days 1, 3, and 5. They were challenged by inhalation at doses of 23.5 mg / kg and 70.5 mg / kg on days 14 (D19) and 21 (D26) after the last sensitization. No active systemic anaphylactic reactions were observed in any of the dose groups of Example 11.

[0280] Example 19 Long-term toxicity test

[0281] ●Rat toxicity test

[0282] A total of 190 SD rats (95 per sex) were used in the experiment and randomly divided into 8 groups according to body weight. Groups 1-4 were the toxicology study groups, and groups 5-8 were the toxicokinetics study groups. Groups 1 and 5 were given sodium chloride injection (0.9%) as negative controls. Groups 2 and 6, 3 and 7, and 4 and 8 were given the actual delivered doses of Example 11 of 17.1, 49.3, and 150.5 mg / kg, respectively, as low, medium, and high dose groups, with inhalation times of 15, 45, and 135 minutes, respectively, once daily for 4 weeks, for a total of 28 administrations, with each administration spaced 24 ± 8 hours apart. The day of the first administration was defined as D1.

[0283] During the administration of the drug from D1 to D28, aerosol samples were taken for aerosol concentration analysis. After the administration of the drug on D1, D7, D14, D21 and D28, samples were taken for aerosol particle size distribution analysis.

[0284] The actual delivered dose and aerosol particle size distribution parameters for the low, medium, and high dose groups are shown in the table below:

[0285] Table 10 Delivery dose and aerosol particle size distribution parameters

[0286] Death or near death: No animal deaths or near death related to Example 11 were observed during the administration and recovery period, in the negative control group, low, medium and high dose groups.

[0287] Toxicological indicators: During the experiment, no abnormal changes related to Example 11 were observed in the negative control group and the dose groups of Example 11 in clinical observation (including general clinical observation, detailed clinical observation and local observation after administration), body weight, food intake, ophthalmological examination, clinicopathology (blood cell count, coagulation function, blood biochemistry and urine analysis), respiratory function, etc.

[0288] Pathological examination:

[0289] No significant abnormalities were observed in the organ weight, gross examination, and microscopic examination of the euthanized animals at the end of the administration period (D29) and the end of the recovery period (D57) as related to Example 11.

[0290] During the experiment, clinical observation (including general clinical observation, detailed clinical observation, and local observation after drug administration) was performed on animals in groups 1-4. This included monitoring body weight, food intake, respiratory function, ophthalmological examination, clinicopathological examination (blood cell count, coagulation function, blood biochemistry, and urinalysis), gross anatomy, organ weighing, and histopathological examination. On days 1 and 28, blood drug concentrations and toxicokinetics were measured in all animals in groups 5-8.

[0291] Fudosteine ​​was not detected in the plasma of animals in the solvent control group after administration on D1 and D28.

[0292] The main TK parameters of fodosteine ​​in each group of animals after administration on D1 and D28 are shown in the table below:

[0293] Table 11 TK Parameters

[0294] Example 11 of fodosteine ​​was administered to SD rats by inhalation at doses of 17.1, 49.3 and 150.5 mg / kg once daily for 28 consecutive days.

[0295] Following administration on days 1 and 28, the exposure (AUC) of fodosteine ​​in female animals of each dose group was determined. last The exposure was higher in males than in females. Exposure increased with increasing dose, and the rate of increase was generally greater than the rate of increase with dose. No significant accumulation was observed overall after repeated administration.

[0296] ●Canine toxicity test

[0297] Forty Beagle dogs (20 per sex) were used in the experiment and randomly divided into four groups according to their body weight. Group 1 animals served as a negative control group, receiving inhalation of 0.9% sodium chloride injection for 120 minutes. Groups 2, 3, and 4 animals received the low, medium, and high doses described in Example 11, with inhalation times of 15, 45, and 120 minutes, respectively, delivering actual doses of 14.9 mg / kg, 44.8 mg / kg, and 122.3 mg / kg. Administered once daily for four weeks, for a total of 28 doses, with the first dose dated D1. The first three animals per sex in groups 1-4 were euthanized at the end of the treatment period (D29), and the last two animals per sex in groups were euthanized after a four-week recovery period (D57).

[0298] During the administration of the drug from D1 to D28, aerosol samples were taken for aerosol concentration analysis. After the administration of the drug on D1, D7, D14, D21 and D28, samples were taken for aerosol particle size distribution analysis.

[0299] During the trial, animals underwent clinical observation (including general clinical observation, detailed clinical observation, and local clinical observation after drug administration), and their weight, food intake, body temperature, electrocardiogram, blood pressure, blood oxygen saturation, ophthalmological examination, clinical pathology (blood cell count, coagulation function, blood biochemistry, and urine analysis), gross anatomy, organ weighing, and histopathological examination were performed. Blood samples were collected on days 1 and 28 for blood drug concentration testing and toxicokinetic analysis.

[0300] The actual delivered dose and aerosol particle size distribution parameters for the low, medium, and high dose groups are shown in the table below:

[0301] Table 12 Actual Delivery Dose and Aerosol Particle Size Distribution Parameters

[0302] Death / Near Death: No animals in groups 1-4 died or were near death during the experiment.

[0303] Toxicological indicators: During the experiment, no changes were observed in the food intake, body temperature, blood oxygen saturation, ophthalmological examination, electrocardiogram, blood pressure, clinicopathology (blood cell count, coagulation function, blood biochemistry and urine analysis), organ weight, gross and microscopic observations of the animals in each treatment group that were related to Example 11.

[0304] Clinical observation: During the trial, no abnormal changes related to Example 11 were observed in the low- and medium-dose groups of animals. In the high-dose group of Example 11, 2 / 10 animals experienced transient coughing on days 7 and 9. Since this occurred in the high-dose group, it was considered possibly related to Example 11. However, it was only transient, and no other abnormal symptoms were observed. Histopathological examination of the respiratory tract was also normal, and it was considered a non-clinical adverse reaction.

[0305] Body weight: During the experiment, no abnormal changes in body weight or weight gain were observed in the low- and medium-dose groups as described in Example 11. In the high-dose group, male animals showed a decrease in body weight (W4) and weight gain, while female animals showed a decrease in weight gain. Compared to the control group at the same time point, male animals (D28) showed a 7.7% decrease in body weight, while female animals showed no decrease. Statistically significant decreases were observed in W4 weight gain for both males and females, by 0.312 and 0.390 kg, respectively. Since the weight loss was less than 10% compared to the negative control group at the same time point and was recoverable during the recovery period, it was considered a non-clinical adverse reaction.

[0306] Toxicokinetics:

[0307] Following administration on days 1 and 28, fodosteine ​​was not detected in the plasma of any animals in the negative control group. Example 11: Animal exposure at each dose group (in AUC) last The exposure rate increased with increasing dose, and the rate of increase in exposure was generally slightly greater than the rate of increase in dose; no significant sex differences were observed; no significant accumulation of fodostein was observed in animals after multiple administrations.

[0308] Table 13 TK Parameters

[0309] Under the experimental conditions, in Example 11, Beagle dogs were repeatedly administered via inhalation at delivery doses of 14.9, 44.8, and 122.3 mg / kg, once daily for 4 weeks, for a total of 28 administrations. No local irritation was observed in any of the groups. No systemic toxicity was observed in the 14.9 and 44.8 mg / kg dose groups. Transient coughing was observed in 2 / 10 of the animals in the 122.3 mg / kg dose group, and a few animals experienced weight loss (less than 10% compared to the negative control group at the same time point). No other abnormalities were observed.

[0310] Example 20 Pharmacodynamic Test

[0311] Eighteen male KM mice were used in this experiment and randomly divided into three groups: a negative control group, a low-dose group (target dose: 15 mg / kg), and a high-dose group (target dose: 30 mg / kg), with six mice per group. The negative control group (inhaled for 30 min), the low-dose group (inhaled for 15 min), and the high-dose group (inhaled for 30 min) were administered the drug via inhalation once daily for three consecutive days (D1-D3). Aerosol samples from D1-D3 were collected for aerosol concentration analysis, and aerosol particle size analysis was performed on D1. Approximately 30 min after inhalation administration on D3, all animals were intraperitoneally injected with phenol red solution (100 mg / kg). All animals were euthanized 30 min after phenol red administration, and tracheal tissue was excised and placed in centrifuge tubes containing NaHCO3 (50 mg / mL) solution. After centrifugation, the supernatant was collected. The OD value was detected at a wavelength of 546 nm using a UV-Vis spectrophotometer, and the amount of phenol red excreted was calculated based on the standard curve.

[0312] In this experiment, D1 to D3 were used for aerosol concentration analysis in Example 11, and D1 samples were used for aerosol particle size analysis. The actual delivery dose and aerosol particle size parameters of Example 11 are shown in Table 14, and the results of the effect of mouse tracheal secretion on the aerosol are shown in Table 15.

[0313] Table 14 Actual delivered dose and aerosol particle size parameters in Example 11

[0314] Note: MMAD: Mass Median Aerodynamic Diameter; GSD: Geometric Standard Deviation; FPF: Fine Particle Fraction, i.e., the percentage of fine particles with MMAD < 5 μm.

[0315] Table 15 Results of the effect of different doses on tracheal erythropoiesis in mice in Example 11

[0316] Phenol red excretion: Compared with the negative control group (7.734±1.867 μg / mL), the low-dose and high-dose groups in Example 11 were 18.383±6.72 μg / mL and 23.897±13.518 μg / mL, respectively, with both mean values ​​increased, and the differences between groups were statistically significant (P≤0.05). This indicates that Example 11, at actual delivery doses of 22.738 mg / kg and 45.474 mg / kg, nebulized inhalation for 15 min and 30 min, respectively, once a day for 3 consecutive days, can increase phenol red excretion in the trachea of ​​mice. Example 21 Aerodynamic particle size distribution

[0317] Install the tray onto the NGI bottom support, ensuring that each collection cup corresponds to its respective position on the bottom support. Pre-cool the assembled impactor and L-shaped connecting tube (artificial trachea) in a cooling device (5°C) for 90 minutes. Connect the impactor to the L-shaped connecting tube, turn on the vacuum pump, connect the flow meter to the L-shaped connecting tube, and adjust the flow control valve to make the gas flow rate at the artificial trachea inlet 15 L / min (±5%). Remove the flow meter.

[0318] Connect the nebulizer to the artificial throat tube, take one sample from Example 11, squeeze it all into the nebulizer, connect the nozzle to the L-shaped connecting tube through the adapter and check the airtightness, turn on the vacuum pump for 15 seconds and then turn on the nebulizer to atomize, turn off the nebulizer after 60 seconds of atomization, wait for 15 seconds, turn off the vacuum pump, and remove the impactor from the cooling device.

[0319] Rinse each collection cup (S1-S7), MOC, adapter (ADP), artificial throat (INP), and nebulizer (DEV) with solvent. Transfer all samples from each device to different volumetric flasks (as shown in Table 16), dilute to the mark with solvent, and mix well. Repeat the same procedure three times.

[0320] Table 16

[0321] The corresponding aerodynamic particle size distribution results are shown in Table 17:

[0322] Table 17 Repeatability Test Results for Different Atomizers - Aerodynamic Particle Size Distribution Results

[0323] Example 22 Delivery rate and total delivery volume

[0324] Delivery rate: Take one sample from Example 11 and fully extrude it into the nebulizer. Connect the nebulizer to the breathing simulator via the mouth adapter, ensuring all connections are sealed. In standard adult mode, turn on the breathing simulator and set its operating time to 1 minute (15 breathing cycles). Start the nebulizer at the beginning of the breathing cycle and stop the nebulizer and breathing simulator after 1 minute. Remove the filter membrane (65mm specification; PP material) into a 250ml beaker, add an appropriate amount of solvent, and sonicate for 10 minutes. Transfer the entire solution to a 200ml volumetric flask (absorb as much solution as possible from the filter membrane), and rinse the beaker with solvent in small amounts several times. Add the washings to the volumetric flask, dilute to the mark with solvent, and shake well. The amount of active material collected by the first filter membrane compared to the collection time is the delivery rate. Repeat the same procedure 3 times.

[0325] Total delivery volume: Replace the filter membrane (65mm specification; PP material) in the filter and continue atomization until the atomizing device no longer releases obvious aerosols. Record the total delivery volume number of times. Remove the filter membrane and place it in a 250ml beaker. Add an appropriate amount of solvent and sonicate for 10 minutes. Transfer the entire solution to a 200ml volumetric flask (absorb as much solution as possible from the filter membrane). Rinse the beaker with solvent in small amounts several times. Add the washing solution to the volumetric flask and dilute to the mark with solvent. Shake well. Accurately measure 1ml of the above solution and place it in a 10ml volumetric flask. Dilute to the mark with solvent and shake well. The total delivery volume is the sum of the amounts collected from all filter papers (the filter papers used to determine the delivery rate and total delivery volume). Repeat the same procedure 3 times.

[0326] Residual amount: Wash the residual solution on the above nebulizer (including nebulizer, mouthpiece and adapter) with solvent, place it in a 250ml beaker, add an appropriate amount of solvent and sonicate for 10 minutes, transfer the solution to a 200ml volumetric flask, and rinse the beaker with solvent in small amounts several times. Add the washing solution to the volumetric flask, dilute to the mark with solvent and shake well; accurately measure 1ml of the above solution, place it in a 10ml volumetric flask, dilute to the mark with solvent and shake well to obtain the solution; repeat the same operation 3 times.

[0327] The delivery rate, total delivery volume, and residual amount are shown in Table 18:

[0328] Table 18 Repeatability Test Results for Different Atomizers - Delivery Rate, Total Delivery, and Residual Amount

[0329] The embodiments of the present invention have been described above by way of example. However, the scope of protection of the present invention is not limited to the above embodiments. Any modifications, equivalent substitutions, improvements, etc., made by those skilled in the art within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A fodosteine ​​pharmaceutical composition, characterized in that, The fodosteine ​​pharmaceutical composition comprises the following components: an active pharmaceutical ingredient, a pH adjuster, and water; the concentration of fodosteine ​​in the composition is 20 mg / ml to 200 mg / ml, and the pH value is 3.0 to 6.0; the active pharmaceutical ingredient is one or more of fodosteine, its pharmaceutically acceptable salts, and its hydrates.

2. The pharmaceutical composition according to claim 1, characterized in that: The pH adjuster is an inorganic base, an inorganic acid, a buffer system formed by an inorganic acid and an inorganic acid salt, an organic acid, or a buffer system formed by an organic acid and an organic acid salt. Preferably, the inorganic acid is selected from one or more of hydrochloric acid, sulfuric acid, and phosphoric acid; Preferably, the inorganic acid salt is selected from one or more of sodium carbonate, sodium bicarbonate, disodium hydrogen phosphate, and sodium dihydrogen phosphate; Preferably, the organic acid is selected from one or more of tartaric acid, lactic acid, citric acid, glacial acetic acid, and malic acid; Preferably, the organic acid salt is selected from sodium citrate; Preferably, the inorganic base is selected from one or more of sodium hydroxide, potassium hydroxide, sodium bicarbonate, and triethylamine; More preferably, the pH adjuster is one or more of hydrochloric acid, sulfuric acid, tartaric acid, and sodium hydroxide.

3. The pharmaceutical composition according to claim 1, characterized in that: pH ranges from 3.0 to 3.4, 3.4 to 4.0, or 4.1 to 6.0, for example, 3.0, 3.4, 3.5, 3.7, 3.8, 4.0, 4.1, 4.5, 5.0, 5.5, or 6.0; Preferably, when the pH adjuster is selected from inorganic acids or buffer systems formed by inorganic acids and inorganic acid salts (e.g., the pH adjuster is hydrochloric acid), the pH value of the fudosteine ​​pharmaceutical composition is 3.0 to 5.0; Preferably, when the pH adjuster is selected from inorganic bases (e.g., sodium hydroxide), the pH value of the fudosteine ​​pharmaceutical composition is greater than 5.0 and does not exceed 6.

0.

4. The pharmaceutical composition according to claim 1, characterized in that: The concentration of the active pharmaceutical ingredient is 50 mg / ml to 150 mg / ml, for example, 50 mg / ml, 80 mg / ml, 100 mg / ml or 150 mg / ml. The concentration refers to the ratio of the mass of the active pharmaceutical ingredient to the volume of the fodosteine ​​pharmaceutical composition.

5. The pharmaceutical composition according to claim 1, characterized in that: The fodosteine ​​pharmaceutical composition described herein does not include one or more of the following: antioxidants, surfactants, and metal chelators; For example, the surfactant is selected from Tween, Span, stearic acid, sodium dodecylbenzene sulfonate or lecithin; For example, the antioxidant is selected from tert-butylhydroquinone, vitamin C, or vitamin E; For example, the metal ion chelating agent is ethylenediaminetetraacetic acid.

6. The pharmaceutical composition according to claim 1, characterized in that: The fodosteine ​​pharmaceutical composition comprises the following components: fodosteine, a pH adjuster, and water, wherein the concentration of fodosteine ​​is 20 mg / ml to 200 mg / ml, and the pH adjuster is selected from inorganic acids or buffer systems formed by inorganic acids and inorganic acid salts (e.g., the pH adjuster is hydrochloric acid). The pH value of the fodosteine ​​pharmaceutical composition is 3.0–5.0; The fodosteine ​​pharmaceutical composition does not contain antioxidants, surfactants, or metal chelators; Alternatively, the fodosteine ​​pharmaceutical composition comprises the following components: fodosteine, a pH adjuster, and water, wherein the concentration of fodosteine ​​is 20 mg / ml to 200 mg / ml, and the pH adjuster is selected from inorganic bases (e.g., sodium hydroxide). The pH value of the fodosteine ​​pharmaceutical composition is greater than 5.0 and does not exceed 6.0; The fodosteine ​​pharmaceutical composition does not contain antioxidants, surfactants, or metal chelators; For example, the fodosteine ​​pharmaceutical composition is any of the following formulations: Prescription 1: Fudosteine ​​100 mg / ml, hydrochloric acid and water for injection, adjust pH to 3.7-3.8; Prescription 2: Fudosteine ​​150 mg / ml, hydrochloric acid and water for injection, adjust pH to 3.7-3.8; Prescription 3: Fudosteine ​​200 mg / ml, hydrochloric acid and water for injection, adjust pH to 3.7-3.8; Prescription 4: Fudosteine ​​80 mg / ml, hydrochloric acid and water for injection, adjust pH to 3.7-3.8; Prescription 5: Fudosteine ​​80 mg / ml, hydrochloric acid and water for injection, adjust pH to 3.7-3.8; Prescription 6: Fudosteine ​​80 mg / ml, hydrochloric acid and water for injection, adjust pH to 3.0; Prescription 7: Fudosteine ​​80 mg / ml, hydrochloric acid and water for injection, adjust pH to 3.5; Prescription 8: Fudosteine ​​80 mg / ml, hydrochloric acid and water for injection, adjust pH to 4.0; Prescription 9: Fudosteine ​​80 mg / ml, hydrochloric acid and water for injection, adjust pH to 4.5; Prescription 10: Fudosteine ​​80 mg / ml, hydrochloric acid and water for injection, adjust pH to 5.0; Prescription 11: Fudosteine ​​80 mg / ml, sodium hydroxide and water for injection, adjust pH to 5.5; Prescription 12: Fudosteine ​​80 mg / ml, sodium hydroxide and water for injection, adjust pH to 6.

0.

7. A pharmaceutical preparation comprising the fodosteine ​​pharmaceutical composition according to any one of claims 1 to 6; Preferably, the pharmaceutical preparation is a nebulized inhalation preparation; Preferably, the pharmaceutical preparation is a single dose of 1.0 ml to 10.0 ml, more preferably 1.0 ml to 5.0 ml, and even more preferably 1.0 ml to 2.5 ml.

8. The use of the fudostein pharmaceutical composition according to any one of claims 1 to 6 in the preparation of medicaments for treating and / or preventing bronchial asthma, chronic wheezing bronchitis, bronchiectasis, pulmonary tuberculosis, pneumoconiosis, chronic obstructive emphysema, atypical mycobacterial disease, pneumonia, and diffuse bronchitis; Preferably, the administration site of the fodosteine ​​pharmaceutical composition is one or more of the following: nose, pharynx, trachea, esophagus, and main bronchus.

9. The preparation process of the fudostein pharmaceutical composition according to any one of claims 1 to 6, comprising the following steps: Simply mix the components of the pharmaceutical composition. Preferably, the preparation method further includes sterilizing the mixed components by filtration; the sterilizing filtration is preferably performed using a polyethersulfone membrane. Preferably, the preparation method further includes packaging the pharmaceutical composition in a container; the container is an ampoule or a vial; the material of the ampoule or vial is selected from glass, polypropylene plastic, polyethylene plastic and polyester; preferably glass ampoules or low-density polyethylene ampoules; the container is filled with nitrogen for protection; More preferably, the pharmaceutical composition is packaged in a container, protected by nitrogen, and then filled and sealed; preferably, the dissolved oxygen after filling and sealing is ≤1.0 mg / L.

10. An atomizing component, characterized in that, The nebulizing assembly includes a nebulizing inhalation device and the fodosteine ​​pharmaceutical composition according to any one of claims 1 to 6; The nebulizing inhalation device is selected from either a compressor-type nebulizing inhalation device or a screen-type nebulizing inhalation device; For example, atomization parameters include: flow rate of 5-30 L / min, atomization time of 30-120 s, pre-cooling time of 60-180 min, and / or pre-cooling temperature of 2-8℃; For example, the aerodynamic particle size distribution of the fudostein drug composition atomization results are as follows: the percentage of fine particles (FPF%) is not less than 40% or is 40% to 70%; the amount of fine particles (FPD) is not less than 10 mg or is 10 mg to 25 mg; and / or, the median mass aerodynamic particle size distribution (MMAD) is not greater than 10 μm or is 1 μm to 10 μm, 3 μm to 6 μm; For example, the delivery rate is not less than 5 mg / min or 5 mg / min to 12 mg / min; the total delivery amount is not less than 20 mg or 20 to 110 mg or 20 to 70 mg; and the residual amount is not more than 150 mg or 90 to 150 mg.

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