Pharmaceutical composition of tricyclic PDE3 / PDE4 dual inhibitor compounds

Abstract

The present invention relates to a pharmaceutical composition of a tricyclic PDE3 / PDE4 dual inhibitor compound and a method for preparing the same, and more particularly to a pharmaceutical composition of the compound of formula (I) or a pharmaceutically acceptable salt thereof, a method for preparing the same and uses thereof. TIFF2023512475000019.tif40170

Description

Technical Field

[0001] Cross-reference to Related Applications In this application, the priority of Chinese Patent Application No. 202010042865.X for invention submitted to the China National Intellectual Property Administration on January 15, 2020 is claimed, and it is incorporated herein by reference in its entirety.

[0002] This application belongs to the field of pharmaceuticals, and relates to a pharmaceutical composition of a tricyclic PDE3 / PDE4 dual inhibitor compound, a method for producing the same, and uses thereof. Specifically, it relates to a pharmaceutical composition of a compound of formula (I) or a pharmaceutically acceptable salt thereof, a method for producing the same, and uses thereof.

Background Art

[0003] Phosphodiesterase (PDE) is a superfamily of enzymes containing 11 families, and each family is involved in different signal transmissions and regulates different physiological processes. Among them, PDE3 is the main phosphodiesterase in human airway smooth muscle (ASM). Inhibiting PDE3 increases the intracellular cAMP concentration and relaxes bronchial smooth muscle. PDE4 plays a major regulatory role in the expression of inflammatory mediators and anti-inflammatory mediators, and PDE4 inhibitors can inhibit the release of harmful mediators from inflammatory cells. Therefore, an inhibitor having an inhibitory effect on both PDE3 and PDE4 has the bronchodilation effect of β-adrenergic receptor agonists and the anti-inflammatory action by inhalation of glucocorticoids. The functional complementarity of the dual targets theoretically has a greater effect than a single target, realizes the therapeutic effect that could only be obtained by conventional combined administration with a single drug, eliminates the disadvantage that the physical and chemical properties of the component drugs in combined administration cannot be fully compatible, makes the administration method easier, and is advantageous for administration at a certain dose.

[0004] Victoria Boswell et al, J.Pharmaco.Experi.Therap. 2006, 318, 840 - 848, and WO200005830 reported compounds RPL554 and RPL565, which have long - term bronchodilation and anti - inflammatory effects. However, due to their physical and chemical properties such as poor solubility and high plasma clearance, they are suitable for inhalation administration. However, it was found from the data that their inhibitory activity against PDE4 was not satisfactory and the anti - inflammatory effect was not good. Therefore, it is necessary to develop compounds with good PDE3 / 4 inhibitory activity.

Chemical formula

Prior art documents

Patent documents

[0005]

Patent Document 1

Non - patent documents

[0006]

Non - patent Document 1

Summary of the invention

[0007] In one aspect of the present application, a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and a surfactant is provided.

Chemical formula

[0008] In some embodiments, the pharmaceutical composition further comprises a metal chelating agent (also referred to as a metal complexing agent).

[0009] In some embodiments, the pharmaceutical composition further comprises a buffering agent.

[0010] In some embodiments, the pharmaceutical composition further comprises a diluent.

[0011] In some embodiments, the pharmaceutical composition further comprises an osmotic pressure regulator.

[0012] In some embodiments, the pharmaceutical composition comprises a compound of formula (I) or a pharmaceutically acceptable salt thereof, a surfactant, and a buffer.

[0013] In some embodiments, the pharmaceutical composition comprises a compound of formula (I) or a pharmaceutically acceptable salt thereof, a surfactant, a buffer, and an osmotic pressure regulator.

[0014] In some embodiments, the pharmaceutical composition comprises a compound of formula (I) or a pharmaceutically acceptable salt thereof, a surfactant, a buffer, and a metal chelating agent.

[0015] In some embodiments, the pharmaceutical composition comprises a compound of formula (I) or a pharmaceutically acceptable salt thereof, a surfactant, a buffer, an osmotic pressure regulator, and a metal chelating agent.

[0016] In some embodiments, the pharmaceutical composition comprises a compound of formula (I) or a pharmaceutically acceptable salt thereof, a surfactant, a buffer, an osmotic pressure regulator, a metal chelating agent, and a diluent.

[0017] In some embodiments, the pharmaceutical composition comprises a compound of formula (I) or a pharmaceutically acceptable salt thereof, a surfactant, and at least one of a buffer, an osmotic pressure regulator, a metal chelating agent, and a diluent.

[0018] In some embodiments, the pharmaceutically acceptable salt is selected from maleate, sulfate, mesylate, and p-toluenesulfonate.

[0019] In some embodiments, the pharmaceutically acceptable salt of the compound of formula (I) has a molar ratio of the compound of formula (I) to the acid ion forming the pharmaceutically acceptable salt of (1:1) to (1:2 ) and may be, for example, 1:1, 1:2.

[0020] In some embodiments, in the pharmaceutical composition, the "compound of formula (I) or a pharmaceutically acceptable salt thereof" may be replaced by the "compound of formula (I)".

[0021] In some embodiments, the pharmaceutical composition comprises a compound of formula (I), a surfactant, a buffer, an osmotic pressure regulator, a metal chelating agent, and a diluent.

[0022] In some embodiments, in the pharmaceutical composition, the concentration of the compound of formula (I) or a pharmaceutically acceptable salt thereof is about 0.001 to about 80 mg / mL calculated as the compound of formula (I), preferably 0.002 to 50 mg / mL, more preferably selected from 0.1 to 20 mg / mL.

[0023] The surfactant of the present application is a pharmaceutically acceptable surfactant, for example, a wetting agent. The surfactant may be a nonionic surfactant, an anionic surfactant, a cationic surfactant, or an amphoteric ionic surfactant. Preferably, the one or more surfactants are nonionic surfactants.

[0024] In some embodiments, the surfactant is selected from one or more of ethylene glycol polyoxyethylene ether, polypropylene glycol alkyl ether, alkyl polyglucoside, octylphenol polyoxyethylene ether, alkylphenol polyoxyethylene ether, glycerin alkyl ester, polyoxyethylene sorbitan fatty acid ester (polysorbate), sorbitan alkyl ester, sorbitan fatty acid ester, cocamide MEA, cocamide DEA, dodecyl dimethylamine oxide, copolymer of polyethylene glycol and polypropylene glycol (poloxamer), ethoxylate of tallow alkylamine (POEA).

[0025] Preferably, the surfactant is selected from one or more of polyoxyethylene sorbitan fatty acid esters (e.g., polysorbate-based), sorbitan fatty acid esters (e.g., span-based).

[0026] In some embodiments, the polyoxyethylene sorbitan fatty acid ester is selected from one or more of polysorbate 20 (polyoxyethylene sorbitan laurate, Tween 20), polysorbate 40 (polyoxyethylene sorbitan monopalmitate), polysorbate 60 (polyoxyethylene sorbitan stearate), polysorbate 80 (polyoxyethylene sorbitan monooleate, Tween 80).

[0027] In some embodiments, the sorbitan fatty acid ester is selected from one or more of sorbitan monolaurate (Span 20), sorbitan monopalmitate, sorbitan monostearate, sorbitan tristearate, sorbitan monooleate.

[0028] More preferably, the surfactant is selected from one or more of polysorbate-based and span-based.

[0029] In some specific embodiments, the surfactant is selected from one or more of polysorbate 20, polysorbate 80, and Span 20.

[0030] In some embodiments, the concentration of the surfactant is about 0.01 to about 8 mg / mL. Typically, the concentration of the surfactant in the pharmaceutical composition is about 0.01 to 5 mg / mL, preferably about 0.02 to 3 mg / mL, more preferably about 0.05 to 2 mg / mL, and even more preferably about 0.1 to 1 mg / mL.

[0031] In some embodiments, in the pharmaceutical composition, the mass ratio of the compound of formula (I) or a pharmaceutically acceptable salt thereof (calculated as the compound of formula (I)) to the surfactant is about 1:200 to 100:1, preferably about 1:150 to 50:1, more preferably about 1:50 to 25:1, and even more preferably about 1:1 to 15:1, for example, about 10:1.

[0032] In some embodiments, the buffer is a pharmaceutically acceptable buffer. The buffer may be any buffer solution suitable for use in an inhalation liquid pharmaceutical composition. The buffer is generally one or more selected from sulfuric acid, hydrochloric acid, sodium hydroxide, citric acid, sodium citrate, lactic acid, sodium lactate, acetic acid, sodium acetate, trisodium phosphate, sodium dihydrogen phosphate, disodium hydrogen phosphate, potassium dihydrogen phosphate, tartaric acid, sodium tartrate, glycine, boric acid, phthalic acid. Preferably, the types of buffers are two or more, and the type is a citrate buffer solution or a phosphate buffer solution, and more preferably the sodium salts of both. The citrate buffer solution includes citric acid, sodium citrate, and mixtures thereof. The phosphate buffer solution includes phosphoric acid, monosodium phosphate (i.e., sodium dihydrogen phosphate), disodium hydrogen phosphate, and mixtures thereof.

[0033] In some embodiments, the buffer is selected from citric acid, citrate (e.g., sodium citrate), tartaric acid, tartrate (e.g., sodium tartrate), phosphoric acid, phosphate (e.g., sodium dihydrogen phosphate, disodium hydrogen phosphate).

[0034] In some embodiments, the buffer is selected from citrate salts (e.g., sodium citrate), tartrate salts (e.g., sodium tartrate), and phosphate salts (e.g., sodium dihydrogen phosphate, disodium hydrogen phosphate).

[0035] In some embodiments, the concentration of the buffer is from about 0.01 to about 50 mg / mL, preferably from about 0.05 to about 40 mg / mL, more preferably from 0.1 to about 25 mg / mL, and even more preferably from 0.5 to about 6 mg / mL.

[0036] In some embodiments, the buffer is used to adjust the pH of the pharmaceutical composition to about 3.0 to about 8.5, preferably about 5 to about 7.

[0037] In some embodiments, the osmotic pressure regulator is generally selected from one or more of simple non-toxic salts such as sodium chloride, potassium chloride, or sugars such as glucose, mannitol, xylitol. In some embodiments, the osmotic pressure regulator is sodium chloride.

[0038] The concentration of the osmotic pressure regulator is determined by the amount necessary to obtain isotonicity, e.g., isotonicity with plasma or lung fluid. The concentration of the osmotic pressure regulator is generally from about 0.01 to about 10 mg / mL, more generally from about 5 to 9 mg / mL.

[0039] In some embodiments, the metal chelating agent is selected from one or more of edetate salts such as edetic acid, disodium edetate, calcium disodium edetate. Preferably it is an edetate salt (e.g., calcium salt, sodium salt), and particularly preferably disodium edetate (EDTA-2Na).

[0040] The concentration of the metal chelating agent is determined by the amount of metal ions that can be introduced in the production of the pharmaceutical composition, and is generally from about 0.01 to about 40 mg / mL, preferably from about 0.01 to about 20 mg / It is mL, more preferably about 0.01 to about 5 mg / mL, and even more preferably about 0.01 to about 2 mg / mL.

[0041] In some embodiments, in the pharmaceutical composition, the diluent may be any pharmaceutically acceptable diluent. The diluent is suitable for inhalation administration. Generally, the diluent is selected from one or more of water, ethanol, and glycerol. Preferably, the diluent is water, and more preferably, the diluent is sterile water.

[0042] In some embodiments, in the pharmaceutical composition, the amount of the diluent used may be an appropriate amount such that the concentration of the compound of formula (I) or a pharmaceutically acceptable salt or additive thereof in the pharmaceutical composition is within a predetermined range.

[0043] In some embodiments, the pharmaceutical composition includes the compound of formula (I) and a polysorbate-based or span-based substance, and further, the polysorbate-based or span-based substance is selected from one or more of polysorbate 20, polysorbate 80, and span 20.

[0044] In some embodiments, the pharmaceutical composition further includes a phosphate, and further, the phosphate may be selected from sodium dihydrogen phosphate or its monohydrate and disodium hydrogen phosphate. In some embodiments, the pharmaceutical composition further includes sodium citrate or sodium tartrate.

[0045] In some embodiments, the pharmaceutical composition includes the compound of formula (I), a mixture of a span-based and a polysorbate-based substance (for example, polysorbate 80, polysorbate 20, span 20), and water.

[0046] In some embodiments, the pharmaceutical composition includes the compound of formula (I), a mixture of a span-based and a polysorbate-based substance (for example, polysorbate 80, polysorbate 20, span 20), sodium citrate or sodium tartrate, and water.

[0047] In some embodiments, the pharmaceutical composition comprises a compound of formula (I), a polysorbate (e.g., polysorbate 80, polysorbate 20), sodium dihydrogen phosphate or its monohydrate, disodium hydrogen phosphate, disodium edetate, and water.

[0048] In some embodiments, the pharmaceutical composition comprises a compound of formula (I), a polysorbate (e.g., polysorbate 80, polysorbate 20), sodium dihydrogen phosphate or its monohydrate, disodium hydrogen phosphate, sodium chloride, disodium edetate, and water.

[0049] In some embodiments, the pharmaceutical composition comprises a compound of formula (I), a polysorbate (e.g., polysorbate 80, polysorbate 20), sodium citrate or sodium tartrate, sodium chloride, and water.

[0050] In some embodiments, the pharmaceutical composition comprises a compound of formula (I), a mixture of a span and a polysorbate (e.g., polysorbate 80, polysorbate 20, span 20), sodium citrate or sodium tartrate, sodium chloride, and water.

[0051] In some embodiments, the pharmaceutical composition comprises a compound of formula (I), a surfactant, a buffer, an osmotic pressure regulator, a metal chelating agent, and a diluent. The concentration of the compound of formula (I) is 0.002 - 50 mg / mL, the concentration of the surfactant is 0.02 - 3 mg / mL, the concentration of the buffer is 0.1 - about 25 mg / mL, the concentration of the osmotic pressure regulator is 5 - 9 mg / mL, and the concentration of the metal chelating agent is 0.01 - about 5 mg / mL.

[0052] In some embodiments, in the pharmaceutical composition, the surfactant, buffer, osmotic pressure regulator, metal chelating agent, and diluent are as defined above.

[0053] In some embodiments, the pharmaceutical composition comprises a compound of formula (I), a surfactant, a buffer, an osmotic pressure regulator, a metal chelating agent, and a diluent. The concentration of the compound of formula (I) is 0.002 to 50 mg / mL, the concentration of the surfactant is 0.02 to 3 mg / mL, the concentration of the buffer is 0.1 to about 25 mg / mL, the concentration of the osmotic pressure regulator is 5 to 9 mg / mL, and the concentration of the metal chelating agent is 0.01 to about 5 mg / mL. The surfactant is one or more of polysorbate 20, polysorbate 80, and span 20. The buffer is one or more of sodium dihydrogen phosphate or its monohydrate, disodium hydrogen phosphate, tartaric acid, and citric acid. The osmotic pressure regulator is sodium chloride. The metal chelating agent is one or more of disodium edetate and calcium disodium edetate. The diluent is water.

[0054] In some embodiments, the pharmaceutical composition comprises a compound of formula (I), polysorbate 80, sodium dihydrogen phosphate or its monohydrate, disodium hydrogen phosphate, sodium chloride, disodium edetate, and water. The concentration of the compound of formula (I) is 0.002 to 50 mg / mL, the concentration of polysorbate 80 is 0.02 to 3 mg / mL, the concentration of sodium dihydrogen phosphate and disodium hydrogen phosphate is 0.1 to 25 mg / mL, the concentration of sodium chloride is 5 to 9 mg / mL, and the concentration of disodium edetate is 0.01 to about 5 mg / mL.

[0055] In some embodiments, in the pharmaceutical composition of the compound of formula (I) or a pharmaceutically acceptable salt thereof, the compound of formula (I) is in a solid form. In some embodiments, the compound of formula (I) is a crystal of the compound of formula (I).

[0056] In some embodiments, in the pharmaceutical composition of the compound of formula (I) or a pharmaceutically acceptable salt thereof, the compound of formula (I) is a product obtained by controlling the particle size of the crystal of the compound of formula (I).

[0057] In some embodiments, the crystal of the compound of formula (I) has diffraction peaks in the X-ray powder diffraction pattern using Cu Kα radiation at 2θ angles of 5.81±0.2°, 13.96±0.2°, 15.01±0.2°, 17.95±0.2°, 24.73±0.2°.

[0058] In some embodiments of the present application, the crystal of the compound of formula (I) has diffraction peaks in the X-ray powder diffraction pattern using Cu Kα radiation at 2θ angles of 5.81±0.2°, 8.38±0.2°, 11.16±0.2°, 13.96±0.2°, 14.47±0.2°, 15.01±0.2°, 17.95±0.2°, 24.73±0.2°, 26.13±0.2°.

[0059] In some embodiments, the crystal of the compound of formula (I) has diffraction peaks in the X-ray powder diffraction pattern using Cu Kα radiation at 2θ angles of 5.81±0.2°, 8.38±0.2°, 11.16±0.2°, 13.96±0.2°, 14.47±0.2°, 15.01±0.2°, 16.76±0.2°, 17.95±0.2°, 20.83±0.2°, 24.73±0.2°, 26.13±0.2°.

[0060] In some embodiments, the crystal of the compound of formula (I) has diffraction peaks in the X-ray powder diffraction pattern using Cu Kα radiation at 2θ angles of 5.81±0.2°, 8.38±0.2°, 9.13±0 .2°, 11.16±0.2°, 11.60±0.2°, 12.82±0.2°, 13.96±0.2°, 14.47±0.2°, 15.01±0.2°, 16.76±0.2°, 17.95±0.2°, 18.91±0.2°, 20.83±0.2°, 24.36±0.2°, 24.73±0.2°, 25.78±0.2°, 26.13±0.2°.

[0061] In some embodiments, the crystal of the compound of formula (I) has an X-ray powder diffraction pattern using Cu Kα radiation that includes 5, 6, 7, 8, 9, 10, 11, 12 or more diffraction peaks at 2θ angles selected from 5.81±0.2°, 8.38±0.2°, 9.13±0.2°, 11.16±0.2°, 11.60±0.2°, 12.82±0.2°, 13.96±0.2°, 14.47±0.2°, 15.01±0.2°, 16.76±0.2°, 17.95±0.2°, 18.91±0.2°, 20.83±0.2°, 24.36±0.2°, 24.73±0.2°, 25.78±0.2°, 26.13±0.2°.

[0062] In some embodiments, the crystal of the compound of formula (I) has an X-ray powder diffraction pattern using Cu Kα radiation that includes 5, 6, 7, 8, 9, 10 or 11 diffraction peaks at 2θ angles selected from 5.81±0.2°, 8.38±0.2°, 11.16±0.2°, 13.96±0.2°, 14.47±0.2°, 15.01±0.2°, 16.76±0.2°, 17.95±0.2°, 20.83±0.2°, 24.73±0.2°, 26.13±0.2°.

[0063] In some embodiments, the crystal of the compound of formula (I) has an X-ray powder diffraction pattern using Cu Kα radiation that includes 5, 6, 7, 8 or 9 diffraction peaks at 2θ angles selected from 5.81±0.2°, 8.38±0.2°, 11.16±0.2°, 13.96±0.2°, 14.47±0.2°, 15.01±0.2°, 17.95±0.2°, 24.73±0.2°, 26.13±0.2°.

[0064] In some embodiments, the peak positions and relative intensities of the diffraction peaks in the XRPD pattern of the crystal of the compound of formula (I) are as shown in Table 1 below.

Table 1

[0065] In some embodiments, the crystal of the compound of formula (I) has an XRPD pattern using Cu Kα radiation as shown in Figure 1.

[0066] In some embodiments, the crystal of the compound of formula (I) has an endothermic peak at 247.70 °C ± 2 °C in the differential scanning calorimetry curve.

[0067] In some embodiments, the crystal of the compound of formula (I) has a differential scanning calorimetry curve as shown in Figure 2.

[0068] In some embodiments, the thermogravimetric analysis curve of the crystal of the compound of formula (I) shows a weight loss of 0.4870% at 155.75 ± 2 °C and a weight loss of 7.287% at 155.75 ± 2 °C to 262.18 ± 2 °C.

[0069] In some embodiments, the crystal of the compound of formula (I) has a thermogravimetric analysis curve as shown in Figure 3.

[0070] In some embodiments, in the pharmaceutical composition, the particle size of the compound of formula (I) or a pharmaceutically acceptable salt thereof is X 50 ≤ 10 μm, preferably 0.1 - 8 μm.

[0071] In some embodiments, in the pharmaceutical composition, the particle size of the compound of formula (I) or a pharmaceutically acceptable salt thereof is X 50 ≤ 5 μm and X 90 ≤ 10 μm.

[0072] The pharmaceutical composition of the present application may be in various forms of preparations formed to be suitable for administration to humans orally or by inhalation, for example, solutions and the like.

[0073] In some embodiments, the pharmaceutical composition of the present application is administered by inhalation.

[0074] In some embodiments, the pharmaceutical composition of the present application is administered by oral or nasal inhalation.

[0075] In some embodiments, the pharmaceutical composition of the present application is an inhalation solution.

[0076] In some embodiments, the pharmaceutical composition of the present application is in the form of a suspension.

[0077] In some embodiments, the pharmaceutical composition of the present application is in the form of an inhalation suspension.

[0078] In another aspect of the present application, there is provided a method for producing the pharmaceutical composition, which includes a process of mixing a surfactant with the compound of formula (I) or a pharmaceutically acceptable salt thereof. Preferably, the production method includes mixing a surfactant with the compound of formula (I) or a pharmaceutically acceptable salt thereof and at least one selected from a metal chelating agent, a buffer, a diluent, and an osmotic pressure regulator. More preferably, in the present application, there is provided a method for producing the pharmaceutical composition, which includes a step of mixing a wetting agent, a buffer, an osmotic pressure regulator, a metal chelating agent, the compound of formula (I) or a pharmaceutically acceptable salt thereof, and a diluent.

[0079] In some embodiments, the method for producing the pharmaceutical composition is as follows: 1) A step of mixing a wetting agent, a buffer, an osmotic pressure regulator, a metal chelating agent, and a diluent to form a solution; 2) A step of mixing the compound of formula (I) or a pharmaceutically acceptable salt thereof with the solution of step 1).

[0080] In some embodiments, the production process further includes step 3) of homogenizing the product of step 2).

[0081] In some embodiments, after the homogenization step, the particle size of the compound of formula (I) or a pharmaceutically acceptable salt thereof is X 50 ≤5 μm and X 90 ≤10 μm.

[0082] In some embodiments, the production process further includes a filling step.

[0083] In another aspect of the present application, there is further provided a method for preventing or treating PDE3- and / or PDE4-related diseases in a mammal, preferably a human, comprising administering to the mammal in need of prevention or treatment a therapeutically effective amount of the pharmaceutical composition.

[0084] In another aspect of the present application, there is further provided the use of the pharmaceutical composition in the manufacture of a drug for preventing or treating PDE3- and / or PDE4-related diseases.

[0085] In another aspect of the present application, there is further provided the use of the pharmaceutical composition for preventing or treating PDE3- and / or PDE4-related diseases.

[0086] In another aspect of the present application, there is further provided the pharmaceutical composition for preventing or treating PDE3- and / or PDE4-related diseases.

[0087] In some embodiments of the present application, the PDE3- and / or PDE4-related diseases are selected from asthma and chronic obstructive pulmonary disease (COPD).

Advantages of the Invention

[0088] The compound of formula (I) of the present application and its pharmaceutical composition have obvious dual inhibitory effects on PDE3 and PDE4, and also have an obvious inhibitory effect on TNF-α in human peripheral blood mononuclear cells (hPBMC). It shows excellent anti-inflammatory effects in a lipopolysaccharide (LPS)-induced rat acute lung injury model. It has high in vivo plasma clearance, low plasma exposure and bioavailability by oral administration, and high safety by local route administration. It has low inhibitory effects on five isoenzymes of cytochrome P450 derived from human liver microsomes (CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP3A4), and there is no risk of drug interaction. It has obvious anti-inflammatory effects such as reducing the total number of white blood cells in BALF, and the minimum effective dose is low. It can reduce the airway resistance index Penh.

[0089] The crystals of the compound of formula (I) of the present application and the crystals of pharmaceutically acceptable salts thereof have advantages in terms of drug activity, pharmacokinetics, bioavailability, hygroscopicity, melting point, stability, solubility, purity, ease of manufacture, etc., and thus meet the requirements regarding the manufacture, storage, transportation, and formulation of drugs. Since the crystal form of the compound of formula (I) has weak hygroscopicity, it is advantageous for the absorption of inhaled drugs.

[0090] The pharmaceutical composition of the compound of formula (I) of the present application has good stability, does not increase impurities, and is within the pharmaceutically acceptable range. It has good dispersibility, no obvious sedimentation of particles, stable pharmacokinetics such as no increase in particle size and being within the range required for effective delivery of inhalation products. Also, the composition has a uniform and appropriate particle size and a fast absorption rate. It has an excellent delivery rate and accurate dosage, a high proportion of aerosol particles that can be inhaled, and a high dosage in the form of fine particles.

[0091] "Explanation of Terms" Unless otherwise specified in the present application, throughout the specification and the appended claims, the term "comprise", similar terms, and their English equivalents, such as "comprises", "comprising", or their equivalents, are understood in an open and inclusive sense of "including... but not limited to", meaning that in addition to the recited elements, components, or steps, other elements, components, or steps not specified may also be included.

[0092] As used throughout the specification, "one embodiment", "an embodiment", "in another embodiment", or "in some embodiments" means that at least one embodiment includes the relevant specific reference elements, structures, or features described in that embodiment. Therefore, the expressions "in one embodiment", "in an embodiment", "in another embodiment", or "in some embodiments" that appear in different places throughout the specification do not necessarily refer to the same embodiment. Also, the specific elements, structures, or features can be combined in any suitable manner in one or more embodiments.

[0093] In addition, the singular limiting term "one" (corresponding to "a", "an", and "the" in English) used in the specification of this application and the appended claims includes cases of multiple objects unless otherwise clearly specified. In other words, in this specification, singular terms include the plural forms of the terms, and vice versa. Therefore, for example, a reaction involving a "catalyst" means a reaction involving one catalyst, or two or more catalysts. In addition, unless otherwise clearly specified, the term "or" is generally used to include the meaning of "and / or".

[0094] The term "treatment" means administering the compounds or formulations described in this application to improve or eliminate a disease or one or more symptoms associated with the disease, and includes the following matters. (i) inhibiting the disease or the state of the disease, i.e., suppressing its progression; (ii) alleviating the disease or the state of the disease, i.e., eliminating the disease or the state of the disease.

[0095] The term "prevention" means administering the compounds or formulations described in this application to prevent a disease or one or more symptoms associated with the disease, and includes preventing the occurrence of a disease or a state of the disease in a mammal, particularly prevention when a mammal prone to the state of the disease has not been diagnosed with the state of the disease.

[0096] The term "therapeutically effective amount" refers to the dosage of the compound of this application that (i) treats or prevents a specific disease, symptom, or disorder, (ii) reduces, improves, or eliminates one or more symptoms of a specific disease, symptom, or disorder, or (iii) prevents or delays the occurrence of one or more symptoms of a specific disease, symptom, or disorder described in this specification. The "therapeutically effective amount" of the compound of this application varies depending on the compound, the state of the disease and its severity, the mode of administration, and the age of the mammal to be treated, but can be determined by those skilled in the art based on their knowledge and the content of this disclosure.

[0097] Generally, the particle size is quantified by measuring the diameter of an equivalent sphere (referred to as the sphere equivalent diameter) characteristic of laser diffraction, for example, using a laser particle size analyzer.

[0098] In the present application, the particle size distribution is represented by the sphere equivalent diameter (VD).

[0099] The term "X 10 " refers to the corresponding particle diameter when the percentage of the cumulative distribution is 10%, which physically means that particles with a diameter less than that account for 10% of the total volume.

[0100] The term "X 50 " is called the volume median diameter and refers to the corresponding particle diameter when the percentage of the cumulative distribution is 50%. Physically, it means that particles with a diameter less than that account for 50% of the total volume.

[0101] The term "X 90 " refers to the corresponding particle diameter when the percentage of the cumulative distribution is 90%, which physically means that particles with a diameter less than that account for 90% of the total volume.

[0102] In this specification, unless otherwise specified, the values of each parameter (including the 2θ angle, reaction conditions, etc.) are all regarded as being modified by the term "about" so as to include errors in the values obtained by measurement, etc. For example, there is an error of ±5% with respect to the described value.

[0103] In this specification, for the purpose of explanation and disclosure, patents, patent applications or existing publications are incorporated by reference in their entirety. These publications can be provided because they were published before the filing date of the present application. The statements regarding the publication dates of these documents or the description of their contents are based on the information known to the applicant, and the applicant does not undertake that the publication dates or the contents of these documents are correct. Moreover, in all target countries, the incorporation of these publications into this specification does not recognize that these publications have become common general knowledge in this field.

Brief Description of Drawings

[0104]

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

Modes for Carrying Out the Invention

[0105] The following specific examples are provided so that those skilled in the art can clearly understand and implement the present application. They should be regarded as illustrative descriptions and typical examples of the present application, rather than limitations on the scope of the present application.

Examples

[0106] Synthesis of Intermediate BB-1

Chemical Formula

Chemical Formula

[0107] Step 1: Synthesis of Compound BB-1-2 Under a nitrogen atmosphere, a mixture of Compound BB-1-1 (21.10 g) and ethyl cyanoacetate (11.00 g, 10.38 mL) was stirred at 100 °C for 16 hours. After the reaction was completed, the mixture was cooled to 70 °C, and ethanol (30 mL) was slowly added dropwise, and a large amount of solid precipitated. It was filtered, and the cake was dried under reduced pressure to obtain Product BB-1-2. 11H NMR (400 MHz, DMSO-d6) δ = 8.26 (t, J = 5.2 Hz, 1H), 6.86 (d, J = 8.0 Hz, 1H), 6.79 (br s, 1H), 6.71 (d, 8.0 Hz, 1H), 4.00 (q, J = 6.8 Hz, 2H), 3.72 (s, 3H), 3.59 (s, 2H), 3.31~3.23 (m, 2H), 2.64 (t, J = 7.2 Hz, 2H), 1.32 (t, J = 6.8 Hz, 3H). MS-ESI m / z: 263.1 [M+H] + 。

[0108] Step 2: Synthesis of Compound BB-1-3 Under a nitrogen atmosphere, phosphoryl trichloride (379.50 g, 230.00 mL) was heated to 85 °C, and compound BB-1-2 (26.00 g) was added in several portions. The reaction mixture was reacted for 2 hours while stirring at 85 °C. After completion of the reaction, most of the phosphoryl trichloride was removed by distillation under reduced pressure. Dichloromethane (200 mL) was added to the residue, and it was washed with water (100 mL × 2). After drying the organic phase over anhydrous sodium sulfate, it was filtered to remove the drying agent and concentrated under reduced pressure. The crude product was obtained, pulped with ethyl acetate (20 mL) and purified to obtain compound BB-1-3. 1 1H NMR (400 MHz, CD3OD) δ = 7.16 (s, 1H), 6.83 (s, 1H), 4.62 (s, 1H), 4.12 (q, J = 6.8 Hz, 2H), 3.86 (s, 3H), 3.35 (d, J = 6.4 Hz, 2H), 2.84 (t, J = 6.4 Hz, 2H), 1.44 (t, J = 6.8 Hz, 3H). MS-ESI m / z: 245.1 [M+H] + 。

[0109] Step 3: Synthesis of Compound BB-1-4 At 0 °C, compound BB-1-3 (1.00 g) was added to 98% concentrated sulfuric acid (12.88 g, 128.69 mmol, 7.00 mL) in several portions. The reaction mixture was stirred at 27 °C for 3 hours. After completion of the reaction, the mixture was added to cold water (15 mL), and then an aqueous sodium hydroxide solution (4 mol / L, 32 mL) was added dropwise to adjust the pH to neutral, followed by extraction with ethyl acetate (100 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered to remove the drying agent, and concentrated under reduced pressure to obtain compound BB-1-4. MS-ESI m / z: 263.1 [M+H] + 。

[0110] Step 4: Synthesis of compound BB-1-5 At 0 °C, sodium (2.42 g) was added to ethanol (80 mL) in several portions. After the mixture was stirred at 28 °C for 30 minutes, compound BB-1-4 (6.90 g) was added to the solution in several portions, and the mixture was stirred at 80 °C for 30 minutes. Subsequently, diethyl carbonate (9.32 g, 9.51 mL) was added all at once, and the mixture was continuously stirred at 80 °C for 5 hours. After completion of the reaction, the mixture was cooled to room temperature, ice water (30 mL) was slowly added, and then the pH was adjusted to neutral with dilute hydrochloric acid (2 mol / L, 53 mL), resulting in the precipitation of a large amount of solid. The solid was filtered to obtain a cake, which was pulped with ethanol (10 mL) and purified to obtain compound BB-1-5. 1 H NMR (400 MHz, DMSO-d6) δ = 11.22 (br s, 1H), 7.35 (s, 1H), 6.95 (s, 1H), 6.22 (s, 1H), 4.09 (q, J = 6.8 Hz, 2H), 3.90 (br s, 2H), 3.83 (s, 3H), 2.89 (br s, 2H), 1.35 (t, J = 6.8 Hz, 3H). MS-ESI m / z: 289.1 [M+H] + 。

[0111] Step 5: Synthesis of compound BB-1-6 At room temperature, compound BB-1-5 (5.00 g) was dissolved in phosphoryl trichloride (30 mL). Under a nitrogen atmosphere, the reaction mixture was stirred at 100 °C for 16 hours. After completion of the reaction, most of the solvent was removed by distillation under reduced pressure. Water (100 mL) was added, and the mixture was extracted with dichloromethane (150 mL × 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered to remove the drying agent, and concentrated under reduced pressure to obtain compound BB-1-6. MS-ESI m / z: 306.9 [M+H] + 。

[0112] Step 6: Synthesis of compound BB-1 At room temperature, compound BB-1-6 (925.67 mg) was dissolved in isopropanol (8 mL), and 2,4,6-trimethylaniline (2.10 g) was added. Under a nitrogen atmosphere, the reaction mixture was stirred at 90 °C for 15 hours. After completion of the reaction, the mixture was cooled to room temperature and concentrated under reduced pressure to obtain a residue, which was then pulped and purified with ethanol (6 mL) to obtain compound BB-1. 1 H NMR (400 MHz, DMSO-d6) δ = 8.85 (br s, 1H), 7.27 (s, 1H), 6.97 (s, 1H), 6.90 (s, 2H), 6.45 (s, 1H), 4.10 (q, J = 6.8 Hz, 2H), 3.90 (t, J = 6.0 Hz, 2H), 3.86 (s, 3H), 2.87 (t, J = 6.0 Hz, 2H), 2.45 (s, 3H), 2.11 (s, 6H), 1.37 (t, J = 6.8 Hz, 3H). MS-ESI m / z: 406.2 [M+H] + 。

[0113] Synthesis of compound BB-4

Chemical Structure

Chemical Structure

[0114] Step 1: Synthesis of compound BB-4-1 At room temperature, compound BB-1 (1.00 g) was dissolved in 2-butanone (35 mL), and 2-(2-bromoethyl)isoindoline-1,3-dione (3.76 g), potassium carbonate (3.07 g), and sodium iodide (2.22 g) were added in this order. Under a nitrogen atmosphere, the reaction mixture was stirred at 85 °C for 72 hours. After completion of the reaction, the mixture was concentrated to remove most of the organic solvent, water (30 mL) was added, and the mixture was extracted with ethyl acetate (25 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered to remove the drying agent, concentrated under reduced pressure, and the residue was purified by flash silica gel column chromatography (eluent: petroleum ether:ethyl acetate = 15:1 to 3:1) to obtain compound BB-4-1. MS-ESI m / z: 579.3 [M+H] + 。

[0115] Step 2: Synthesis of compound BB-4 At room temperature, compound BB-4-1 (500.00 mg) was dissolved in chloroform (3 mL) and ethanol (3 mL), and hydrazine hydrate (152.67 mg, purity 85%) was added. Under a nitrogen atmosphere, the mixture was stirred at 28 °C for 16 hours. After completion of the reaction, the mixture was concentrated to remove most of the organic solvent, water (15 mL) was added, and the mixture was extracted with dichloromethane (15 mL × 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered to remove the drying agent, and concentrated under reduced pressure to obtain compound BB-4. 1 H NMR (400 MHz, DMSO-d6) δ = 6.95 (s, 1H), 6.85 (br s, 2H), 6.66 (s, 1H), 5.31 (s, 1H), 4.14 (t, J = 6.8 Hz, 2H), 4.05 (q, J = 6.8 Hz, 2H), 3.91 (t, J = 6.4 Hz, 2H), 3.62 (s, 3H), 2.90~2.86 (m, 4H), 2.22 (s, 3H), 1.95 (br s, 6H), 1.33 (t, J = 6.8 Hz, 3H). MS-ESI m / z: 449.2 [M+H] + 。

[0116] Example 1: Production of the compound of formula (I) At 20 °C, 5-hydroxy-3-methyl-1,2,3-triazole-4-carboxylic acid (18.50 mg) was dissolved in DCM (1 mL), HATU (8.80 mg) and triethylamine (57.40 μL) were added, and the mixture was stirred for 2 hours. Subsequently, compound BB-4 (50 mg ) was added, and stirring was continued for 16 hours while maintaining the temperature. The mixture was diluted to 10 mL with DCM, washed with water (30 mL × 3), dried over anhydrous sodium sulfate, filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to evaporate the solvent to obtain a crude product. The crude product was purified by prep-HPLC to obtain the target compound of formula (I) as a yellow solid. 1 H NMR (400 MHz, CD3OD) δ = 6.94 (s, 2H), 6.87 (s, 1H), 6.77 (s, 1H), 5.52 (s, 1H), 4.48 (t, J = 6.0 Hz, 2H), 4.15 (s, 3H), 4.12~4.08 (m, 2H), 4.01 (t, J = 6.0 Hz, 2H), 3.87 (t, J = 6.0 Hz, 2H), 3.69 (s, 3H), 2.94 (t, J = 6.0 Hz, 2H), 2.29 (s, 3H), 2.06 (s, 6H), 1.41 (t, J = 6.8 Hz, 3H). MS m / z: 574.1 [M + H] + 。

[0117] Example 2: Preparation of crystals of the compound of formula (I) 50 mg of the compound of formula (I) was added to a 4 mL glass bottle, 1 mL of absolute ethanol and 0.2 mL of water were added, the temperature was raised to 40 °C and stirred for 48 hours. It was cooled to room temperature by natural cooling, centrifuged to obtain a solid, and vacuum dried to obtain 46 mg of solid crystals. The XRPD pattern, DSC curve and TGA curve are as shown in Figures 1, 2 and 3 respectively.

[0118] Examples 3 to 36: Manufacturing process: 1) Each additive was added to a preparation tank, stirred to clarify it, and an additive solution was obtained. 2) The compound of formula (I) was added to the additive solution prepared above, and stirred to obtain a uniform suspension. 3) Using a device such as a high-pressure homogenizer, microjet or sand mill, adjust the particle size of the compound of formula (I) in the formulation to X 50 ≤ 5 μm and X 90 ≤ 10 μm. 4) A product was obtained.

[0119] The dosage of each additive and the product are as shown in Table 2 below.

Table 2-1

Table 2-2

[0120] Experimental Example 1: Study on the solid stability of the crystals of the compound of formula (I) High performance liquid chromatography (HPLC): Refer to the following for the composition of the HPLC method. Column: Zorbax SB C-18, 4.6 mm × 150 mm, 5 μm (PDS-HPLC-007). Mobile phase A: 0.1% TFA in water. Mobile phase B: 100% ACN. Sample preparation: The sample was dissolved in a mixed solvent of acetonitrile and water (acetonitrile: water = 50:50 (V:V)).

[0121] Static method for examining solid stability: The stability of the compound when standing under the following conditions was examined, and samples were taken at different times to measure the content. Approximately 5 mg of the crystals of the compound of formula (I) produced in Example 2 was accurately weighed and placed in a dry and clean glass bottle. As a double sample, the thinly spread one was used as the formal test sample, and under the stress factor test conditions ((60 °C), (relative humidity 92.5%), light (total illuminance 1.2 × 10 6 Lux·hr / near ultraviolet region 200 W·hr / m 2) It was left standing at (40 °C, relative humidity 75%), (60 °C, relative humidity 75%) so that the sample was completely exposed, covered with aluminum foil, and small holes were made. Samples were taken and analyzed at 5 days, 10 days, 1 month, 2 months, and 3 months. The samples left standing under light (visible light 1,200,000 Lux, ultraviolet 200 W) conditions were completely exposed at room temperature. Refer to Table 3 for the experimental results.

Table 3

[0122] As can be seen from the above, the crystals of the compound of formula (I) in the present application have good stability under high temperature, high humidity or light conditions, and no impurities increased during the test.

[0123] Experimental Example 2: Study on the hygroscopicity of the crystals of the compound of formula (I) Apparatus model: SMS DVS Advantage dynamic vapor sorption measurement apparatus Measurement conditions: The sample (10 - 20 mg, crystals produced in Example 3) was placed on a DVS sample tray for measurement.

[0124] The detailed DVS parameters are as follows. Temperature: 25 °C. Equilibrium: dm / dt = 0.01% / min (minimum 10 min, maximum 180 min). Drying: Dried for 120 minutes under 0% RH. RH (%) measurement step: 10%. RH (%) measurement step range: 0% - 90% - 0%. The final dynamic vapor sorption (DVS) curve is as shown in Figure 4.

[0125] As can be seen from Figure 4, the crystals of the compound of formula (I) in the present application have low hygroscopicity.

[0126] Experimental Example 3: In vitro measurement of the inhibitory activity of the compound against PDE3A enzyme Experimental purpose: Measure the expression of AMP / GMP by fluorescence polarization, that is, track the binding of the AMP / GMP antibody to show enzyme activity.

[0127] Reagent: Experimental buffer solution: 10 mM Tris-HCl (pH 7.5), 5 mM MgCl2, 0 .01% Brij 35, 1 mM dithiothreitol (DTT), 1% DMSO.

[0128] Enzyme: Recombinant human PDE3A (gene accession number NM_000921, amino acid 669 terminus) with an N-terminal GST tag was expressed in the baculovirus of Sf9 insect cells, and the molecular weight was 84 kDa.

[0129] Substrate: 1 μM cAMP. Measurement: Transcreener® AMP2 / GMP2 antibody and AMP2 / GMP2 AlexaFluor633 were traced.

[0130] Procedure: 1. Recombinant human PDE3A enzyme and substrate (1 μM cAMP) were each dissolved in freshly prepared experimental buffer solution. 2. The PDE3A enzyme buffer solution was transferred to the reaction well. 3. Using an acoustic method (echo 550, accuracy nanoliter), a compound dissolved in 100% DMSO was added to the reaction well containing the PDE3A enzyme buffer solution, and incubated at room temperature for 10 minutes. 4. The substrate buffer solution was added to the reaction well to initiate the reaction. 5. Incubated at room temperature for 1 hour. 6. The measurement mixture (Transcreener® AMP2 / GMP2 antibody and AMP2 / GMP2 AlexaFluor633 tracing) was added to stop the reaction, and incubated for 90 minutes while mixing gently. The measurement range of fluorescence polarization was Ex / Em = 620 / 688.

[0131] Data analysis: Based on the AMP / GMP standard curve, the percent enzyme activity was calculated from the fluorescence polarization signal relative to the DMSO control using software Excel and converted to nM. Curve fitting was performed using GraphPad Prism (for medical graphing). Experimental results are shown in Table 4.

[0132] Experimental Example 4: In vitro measurement of the inhibitory activity of compounds against the PDE4B enzyme Experimental objective: To measure the expression of AMP / GMP by fluorescence polarization, thus tracking the binding of AMP / GMP antibodies and thus indicating enzyme activity.

[0133] reagent: Experimental buffer solution: 10mM Tris-HCl (pH 7.5), 5mM MgCl2, 0.01% Brij 35, 1mM DTT, 1% DMSO.

[0134] Enzyme: Recombinant human PDE4B (gene accession number NM_002600, amino acid 305 end) was expressed in baculovirus in Sf9 insect cells with an N-terminal GST tag and had a molecular weight of 78 kDa.

[0135] Substrate: 1 μM cAMP. Measurements: Transcreener® AMP2 / GMP2 antibody and AMP2 / GMP2 AlexaFluor633 were tracked.

[0136] procedure: 1. Recombinant human PDE4B enzyme and substrate (1 μM cAMP) were dissolved in freshly prepared experimental buffer. 2. The PDE4B enzyme buffer solution was transferred to the reaction well. 3. Using an acoustic technique (echo 550, nanoliter precision), the PDE4B enzyme was detected. Compounds dissolved in 100% DMSO were added to reaction wells containing basic buffer solution and incubated at room temperature for 10 minutes. 4. Substrate buffer solution was added to the reaction well to initiate the reaction. 5. Incubate at room temperature for 1 hour. 6. The reaction was stopped by adding the measurement mixture (Transcreener® AMP2 / GMP2 antibody and AMP2 / GMP2 AlexaFluor633 tracer), and incubated for 90 minutes while mixing slowly. The measurement range of fluorescence polarization was Ex / Em = 620 / 688.

[0137] Data analysis: Based on the AMP / GMP standard curve, the percentage of enzyme activity relative to the DMSO control was calculated from the fluorescence polarization signal using the software Excel and converted to nM. Curve fitting was performed using GraphPad Prism (for medical graph creation).

[0138] Refer to Table 4 for the experimental results.

Table 4

[0139] The active ingredient of the pharmaceutical composition of the present application has an obvious dual inhibitory effect on PDE3 and PDE4.

[0140] Experimental Example 5: Pharmacokinetic study in beagle dogs In this study, male beagle dogs were used as test animals, and the plasma drug concentrations of the compound of formula (I) at different time points after intravenous injection or forced oral administration to beagle dogs were quantitatively measured by LC-MS / MS method, so as to evaluate the in vivo pharmacokinetic characteristics of the compound of formula (I) in beagle dogs.

[0141] A clear solution of the compound of formula (I) was injected into the bodies of two beagle dogs weighing 10 - 12 kg via the cephalic vein or the great saphenous vein, and the clear solution of the compound of formula (I) was forcibly orally administered to two beagle dogs weighing 10 - 12 kg (fasted overnight). All animals were bled approximately 500 μL from the peripheral vein at 0.0333 h, 0.0833 h, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, and 24 h after administration and transferred to anticoagulant centrifuge tubes containing 0.85 - 1.15 mg of K2EDTA·2H2O, and centrifuged at 3000 g for 10 minutes at 4°C to collect plasma. The blood drug concentration was measured by the LC-MS / MS method, and the relevant pharmacokinetic parameters were calculated by the non-compartmental model × linear trapezoidal method using the pharmacokinetic software WinNonlin (trademark) Version 6.3 (Pharsight Corporation, Mountain View, California).

Table 5

[0142] The active ingredient of the pharmaceutical composition of the present application has a high in vivo plasma clearance and low plasma exposure and bioavailability upon oral administration.

[0143] Experimental Example 6: Inhibitory effect on the isoenzymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP3A4) of cytochrome P450 derived from human liver microsomes Phenacetin (CYP1A2), Diclofenac (CYP2C9), (S)-Mephenytoin (CYP2C19), Dextromethorphan (CYP2D6), and Midazolam (CYP3A4), which are five specific probe substrates for five CYP isoenzymes, were incubated with human liver microsomes and the compound of formula (I), respectively. Nicotinamide adenine dinucleotide phosphate reduced form (NADPH) was added to initiate the reaction. After the reaction was completed, the samples were processed and the concentrations of five metabolites, Acetaminophen, 4’-Hydroxydiclofenac, 4’-Hydroxymephenytoin, Dextrorphan, and 1’-Hydroxymidazolam, which were generated from the specific substrates, were quantitatively measured by liquid chromatography tandem mass spectrometry (LC-MS / MS) method, and the corresponding half inhibitory concentration (IC 50 ) was calculated.

Table 6

[0144] The active ingredient of the pharmaceutical composition of the present application has a low inhibitory effect on five isoenzymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP3A4) of cytochrome P450 derived from human liver microsomes.

[0145] Experimental Example 7: Pharmacodynamic study in a rat model of acute lung injury induced by passive smoking Experimental animals: Male SPF-standard Sprague-Dawley rats (provided by Shanghai Slack Experimental Animal Co., Ltd.) were used, and the body weight was about 200 g.

[0146] Experimental procedure: 1. After the animals were accepted into the facility, they were bred for one week to adapt to the environment and then randomly divided into 6 groups according to their body weight. 2. On the 1st to 3rd days of the experiment, in each group, the target compound was atomized and administered for 30 minutes. Then, the animals in the model group and each compound treatment group were exposed to passive smoking for 1 hour, and at 4-hour intervals, they were exposed to passive smoking for 1 hour again. They were exposed to tobacco smoke twice a day for 3 consecutive days. The control group animals were exposed to air indoors. 3. On the 4th day of the experiment, in each group, the target compound was atomized and administered for 30 minutes. The animals in the model group and each compound treatment group were exposed to 150 μg / mL of LPS by atomization inhalation for 15 minutes. 3 hours after the start of atomization administration, they were exposed to passive smoking for 1 hour. The animal lung function (Penh, F) was measured, and after the animals were euthanized with CO2, the alveolar lavage fluid was collected for cell counting.

[0147] 4. Administration treatment Administration method: Using an atomization administration device for whole-body exposure, the test compound and the reference compound were atomized and administered for 30 minutes at the maximum atomization rate (about 12 mL).

[0148] Administration frequency: Atomized and administered for 30 minutes before passive smoking every morning or the solvent was administered, and it was administered before LPS atomization inhalation on the 4th day.

[0149] 5. Measurement of pharmacodynamic endpoints (1) Total number of white blood cells in BALF (alveolar lavage fluid). (2) Measurement of lung function under methacholine (Mch) induction (airway resistance index Penh).

Table 7

[0150] Refer to Figures 5 and 6 for the experimental results.

[0151] As shown in Figures 5 and 6, the active ingredient of the pharmaceutical composition of the present application can effectively reduce the number of white blood cells in the alveolar lavage fluid and the airway resistance index Penh.

[0152] Experimental Example 8: In Vitro Measurement of the Inhibitory Activity of Compounds against TNF-α in Human Peripheral Blood Mononuclear Cells Experimental Purpose: To show the anti-inflammatory activity at the cellular level of the test compound based on the level of TNF-α in human peripheral blood mononuclear cells (hPBMC).

[0153] Procedure: 1. Whole blood was collected from healthy subjects and anticoagulated with an EDTA anticoagulation tube. 2. PBMCs were separated by Ficoll density gradient centrifugation, counted, and the cell concentration was adjusted to 2×10 6 / mL. 3. 2×10 5 cells and 1 ng / mL of LPS were added to each well of a U-bottom 96-well plate. The compound of formula (I) was prepared in DMSO solutions at concentrations of 100 μM, 10 μM, 1 μM, 100 nM, 10 nM, 1 nM, 100 pM, and 10 pM respectively, and each well had a reaction system of 200 μL. 4. After culturing for 24 hours, the supernatant was collected. 5. The TNF-α level in the supernatant was measured by ELISA, and an inhibition curve was fitted with the software Graphpad Prism to calculate the IC 50 .

[0154] Refer to Table 8 for the experimental results.

Table 8

[0155] Therefore, the active ingredient of the pharmaceutical composition of the present application can exhibit effective anti-inflammatory activity and has an obvious inhibitory effect on TNF-α in human peripheral blood mononuclear cells (hPBMC).

[0156] Experimental Example 9: Stability Test The product obtained in Example 28 was left standing for 6 months under accelerated conditions (40°C ± 2°C / RH 25% ± 5%) and long-term conditions (30°C ± 2°C / RH 65% ± 5%). Refer to Table 10 for the results.

Table 9

[0157] As can be seen from the above, the pharmaceutical composition of the present application exhibits good stability under accelerated conditions and long-term conditions, with no obvious increase in impurities and no increase in particle size.

Claims

1. A pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and a surfactant. 【Chemical Formula 1】

2. The pharmaceutical composition according to claim 1, further comprising a metal chelating agent.

3. The pharmaceutical composition according to claim 1 or 2, further comprising a buffering agent.

4. The pharmaceutical composition according to any one of claims 1 to 3, further comprising an osmotic pressure regulator.

5. The pharmaceutical composition according to any one of claims 1 to 4, further comprising a diluent.

6. The pharmaceutical composition according to any one of claims 1 to 5, comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, a surfactant, and at least one of a buffering agent, an osmotic pressure regulator, a metal chelating agent, and a diluent.

7. The surfactant is a nonionic surfactant, or the surfactant is selected from one or more of ethylene glycol polyoxyethylene ether, polypropylene glycol alkyl ether, alkyl polyglucoside, octylphenol polyoxyethylene ether, alkylphenol polyoxyethylene ether, glycerin alkyl ester, polyoxyethylene sorbitan fatty acid ester, sorbitan alkyl ester, sorbitan fatty acid ester, cocamide MEA, cocamide DEA, dodecyldimethyl amine oxide, a copolymer of polyethylene glycol and polypropylene glycol, and ethoxylates of tallow alkylamines, or the surfactant is selected from one or more of polysorbate-based and span-based surfactants, or the surfactant is selected from one or more of polysorbate 20, polysorbate 80, and span 20, or the concentration of the surfactant is about 0.01 to about 8 mg / mL, the pharmaceutical composition according to any one of claims 1 to 6.

8. The buffering agent is selected from one or more of sulfuric acid, hydrochloric acid, sodium hydroxide, citric acid, sodium citrate, lactic acid, sodium lactate, acetic acid, sodium acetate, trisodium phosphate, sodium dihydrogen phosphate, disodium hydrogen phosphate, potassium dihydrogen phosphate, tartaric acid, sodium tartrate, glycine, boric acid, and phthalic acid, or the buffering agent is selected from citric acid, citrate, tartaric acid, tartrate, phosphoric acid, and phosphate, or the buffering agent is selected from citrate, tartrate, and phosphate, or the concentration of the buffering agent is about 0.01 to about 50 mg / mL. Or, the pharmaceutical composition according to claim 3, wherein the buffer is used to adjust the pH of the pharmaceutical composition to about 3.0 to about 8.

5.

9. The pharmaceutical composition according to claim 4, wherein the osmotic pressure regulator is selected from one or more of sodium chloride, potassium chloride, glucose, mannitol, and xylitol.

10. The pharmaceutical composition according to claim 2, wherein the metal chelating agent is selected from one or more of edetic acid, disodium edetate, and calcium disodium edetate.

11. The pharmaceutical composition according to claim 5, wherein the diluent is selected from one or more of water, ethanol, and glycerol.

12. A pharmaceutical composition comprising a compound of formula (I), a surfactant, a buffer, an osmotic pressure regulator, a metal chelating agent, and a diluent, wherein the concentration of the compound of formula (I) is 0.002 to 50 mg / mL, the concentration of the surfactant is 0.02 to 3 mg / mL, the concentration of the buffer is 0.1 to about 25 mg / mL, the concentration of the osmotic pressure regulator is 5 to 9 mg / mL, and the concentration of the metal chelating agent is 0.01 to about 5 mg / mL, according to any one of claims 1 to 11.

13. A pharmaceutical composition comprising a compound of formula (I), polysorbate 80, sodium dihydrogen phosphate or its monohydrate, disodium hydrogen phosphate, sodium chloride, disodium edetate, and water, and optionally, the concentration of the compound of formula (I) is 0.002 to 50 mg / mL, the concentration of polysorbate 80 is 0.02 to 3 mg / mL, the concentration of sodium dihydrogen phosphate and disodium hydrogen phosphate is 0.1 to 25 mg / mL, the concentration of sodium chloride is 5 to 9 mg / mL, and the concentration of disodium edetate is 0.01 to about 5 mg / mL, according to any one of claims 1 to 12.

14. The compound of formula (I) is a crystal of the compound of formula (I), and the X-ray powder diffraction pattern of the crystal using Cu Kα radiation has 5, 6, 7, 8, 9, 10 or 11 diffraction peaks at 2θ angles selected from 5.81 ± 0.2°, 8.38 ± 0.2°, 11.16 ± 0.2°, 13.96 ± 0.2°, 14.47 ± 0.2°, 15.01 ± 0.2°, 16.76 ± 0.2°, 17.95 ± 0.2°, 20.83 ± 0.2°, 24.73 ± 0.2°, 26.13 ± 0.2°, or the X-ray powder diffraction pattern using Cu Kα radiation has diffraction peaks at 2θ angles of 5.81 ± 0.2°, 13.96 ± 0.2°, 15.01 ± 0.2°, 17.95 ± 0.2°, 24.73 ± 0.2°, or 2θ angles of 5.81 ± 0.2°, 8.38 ± 0.2°, 11.16 ± 0.2°, 13.96 ± 0.2°, 14.47 ± 0.2°, 15.01 ± 0.2°, 16.76 ± 0.2°, 17.95 ± 0.2°, 20.83 ± 0.2°, 24.73 ± 0.2°, 26.13 ± 0.2°; the pharmaceutical composition according to any one of claims 1 to 13.

15. The particle size of the compound of formula (I) or a pharmaceutically acceptable salt thereof is X ５０ ≦ 10 μm, or the particle size of the compound of formula (I) or a pharmaceutically acceptable salt thereof is X ５０ ≦ 5 μm and X ９０ ≦ 10 μm, The pharmaceutical composition according to any one of claims 1 to 14

16. The mass ratio of the compound of formula (I) or a pharmaceutically acceptable salt thereof to the surfactant is about 1:200 to 100:1, preferably about 1:150 to 50:1, more preferably about 1:50 to 25:1, and even more preferably about 1:1 to 15:1 (the mass of the compound of formula (I) or a pharmaceutically acceptable salt thereof is calculated based on the compound of formula (I)); the pharmaceutical composition according to any one of claims 1 to 15.

17. The pharmaceutical composition according to any one of claims 1 to 16, wherein the pharmaceutical composition is in the form of a suspension.

18. A method for producing the pharmaceutical composition according to claim 6, comprising mixing a surfactant with a compound of formula (I) or a pharmaceutically acceptable salt thereof and at least one selected from a metal chelating agent, a buffer, a diluent, and an osmotic pressure regulator.

19. A medicament for preventing or treating PDE3 and / or PDE4-related diseases in mammals, comprising the pharmaceutical composition according to any one of claims 1 to 17, and optionally, the PDE3 and / or PDE4-related diseases are selected from asthma and chronic obstructive pulmonary disease; the medicament.

20. Use of the pharmaceutical composition according to any one of claims 1 to 17 in the manufacture of a medicament for preventing or treating a PDE3 and / or PDE4-related disease, optionally, wherein the PDE3 and / or PDE4-related disease is selected from asthma and chronic obstructive pulmonary disease.

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