Pharmaceutical composition containing isoquinolinone compound and preparation method thereof

Abstract

The invention relates to a pharmaceutical composition containing an isoquinolinone compound and a preparation method of the pharmaceutical composition. In particular, the present disclosure provides a pharmaceutical composition comprising 9, 10-dimethoxy-2-[[2-(2-oxo-imidazolin-1-yl)-ethyl]-(2, 4, 6-trimethyl-phenyl)-amino]-6, 7-dihydro-pyrimidinyl [6, 1-a]-isoquinolin-4-one or a pharmaceutically acceptable salt thereof, and a polysorbate 80. The pharmaceutical composition shows good physical and chemical stability after being stored for several months.

Description

A pharmaceutical composition containing isoquinolinone compounds and a preparation method thereof

[0001] The present invention belongs to the field of pharmaceutical preparations, and relates to a pharmaceutical composition comprising an isoquinolinone compound and a preparation method thereof.

[0002] The development of new molecules with both PDE3 and PDE4 inhibitory activity will have the bronchodilator effect of β-adrenergic receptor agonists and the anti-inflammatory effect of inhaled glucocorticoids. The dual-targeting function complements each other and has better efficacy than a single target. For example, RPL554 (9,10-Dimethoxy-2-(2,4,6-trimethylphenylimino)-3-(N-carbamoyl-2-aminoethyl)-3,4,6,7-tetrahydro-2H-pyrimido[6,la]isoquinolin-4-one) is a PDE3 / PDE4 dual-target inhibitor, disclosed in WO00 / 58308. Recent Phase II clinical data showed that it can significantly improve bronchiectasis and symptoms in patients with COPD. At the same time, the drug is well tolerated and no obvious adverse events occur, such as heart problems, nausea and diarrhea, which are all mild. The safety of the drug and its "limited systemic exposure" are encouraging.

[0003] 9,10-dimethoxy-2-[[2-(2-oxo-imidazolin-1-yl)-ethyl]-(2,4,6-trimethyl-phenyl)-amino]-6,7-dihydro-pyrimidinyl[6,1-a]-isoquinolin-4-one is a new type of PDE3 / 4 with better PDE3 and PDE4 inhibitory activity.

[0004] In order to make new PDE3 / 4 inhibitors suitable for administration and to maintain stability during storage and subsequent use and to exert better effects, it is necessary to develop pharmaceutical (preparation) compositions that are more suitable for administration.

[0005]

[0006] The present disclosure provides a pharmaceutical composition comprising an active ingredient 9,10-dimethoxy-2-[[2-(2-oxo-imidazolin-1-yl)-ethyl]-(2,4,6-trimethyl-phenyl)-amino]-6,7-dihydro-pyrimidinyl[6,1-a]-isoquinolin-4-one or a pharmaceutically acceptable salt thereof, and polysorbate 80.

[0007] The concentration of the active ingredient in the pharmaceutical composition described in the present disclosure is 0.01 to 40 mg / mL, including but not limited to 0.01 mg / ml, 0.02 mg / ml, 0.03 mg / ml, 0.04 mg / ml, 0.05 mg / mL, 0.1 mg / mL, 0.3 mg / mL, 0.5 mg / mL, 0.7 mg / mL, 0.9 mg / mL, 1.1 mg / mL, 1.3 mg / mL, 1.5 mg / mL, 1.7 mg / mL, 1.9 mg / mL, 2.1 mg / mL, 2.3 mg / mL, 2.5 mg / mL, 2.7 mg / mL, 2.9 mg / mL, 3.1 mg / mL, 3.3 mg / mL, 3.5 mg / mL, 3.7 mg / mL, 3.9 mg / mL, 4.1 mg / mL, 4.3 mg / mL, 4.5 mg / mL, 4.7 mg / mL, 4.9 mg / mL, 5.1 mg / mL, 5.3 mg / mL, 5.5 mg / mL, 5.7 mg / mL, 5.9 mg / mL, 6.1 mg / mL, 6.3 mg / mL, 6.5 mg / mL, 6.7 mg / mL, 6.9 mg / mL, 7.1 mg / mL, 7.3 mg / mL, 7.5 mg / mL, 7.7 mg / mL, 7.9 mg / mL, 8.1 mg / mL, 8.3 mg / mL, 8.5 mg / mL, 8.7 mg / mL, 8.9 mg / mL, 9.1 mg / mL, 9.3 mg / mL, 9.5 mg / mL, 9.7 mg / mL, 9.9 mg / mL, 10.1 mg / mL, 10.3 mg / mL, 10.5 mg / mL, 10.7 mg / mL, 10.9 mg / mL, 11.1 mg / mL, 11.3 mg / mL, 11.5 mg / mL, 11.7 mg / mL, 11.9 mg / mL, 12.1 mg / mL, 12.3 mg / mL, 12.5 mg / mL, 12.7 mg / mL, 12.9 mg / mL, 13.1 mg / mL, 13.3 mg / mL, 13.5 mg / mL, 13.7 mg / mL, 13.9 mg / mL, 14.1 mg / mL, 14.3 mg / mL, 14.5 mg / mL, 14.7 mg / mL, 14.9 mg / mL, 15.1 mg / mL, 15.3 mg / mL, 15.5 mg / mL, 15.7 mg / mL, 15.9 mg / mL, 16.1 mg / mL, 16.3 mg / mL, 16.5 mg / mL, 16.7 mg / mL, 16.9 mg / mL, 17.1 mg / mL, 17.3 mg / mL, 17.5 mg / mL, 17.7 mg / mL, 17.9 mg / mL, 18.1 mg / mL, 18.3 mg / mL, 18.5 mg / mL, 18.7 mg / mL, 18.9 mg / mL, 19.1 mg / mL, 19.3mg / mL, 19.5mg / mL, 19.7mg / mL, 19.9mg / mL, 20.0mg / mL or any value between two values.

[0008] In some embodiments, the concentration of the active ingredient in the pharmaceutical composition is 0.05 to 20 mg / mL.

[0009] Further, the pharmaceutical composition of the present disclosure also contains a buffer, and the buffer is selected from a citrate buffer, a phosphate buffer or an acetate buffer. In some embodiments, the buffer is selected from a phosphate buffer. In other embodiments, the buffer is selected from a sodium dihydrogen phosphate-disodium hydrogen phosphate buffer. In other embodiments, the buffer is selected from a sodium acetate-acetate buffer.

[0010] In an alternative embodiment, the concentration of the buffer is 1 mM to 50 mM, including but not limited to 1 mM, 2 mM, 3 mM, 4 mM, 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 11 mM, 12 mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, 20 mM, 21 mM, 22 mM, 23 mM, 24 mM, 47mM, 48mM, 49mM, 50mM or any value between any two values.

[0011] In some embodiments, the concentration of the buffer is 5 to 30 mM.

[0012] On the other hand, some embodiments provide pharmaceutical compositions in which the concentration of polysorbate 80 is 0.05 to 5.0 mg / mL, including but not limited to 0.05 mg / mL, 0.06 mg / mL, 0.07 mg / mL, 0.08 mg / mL, 0.09 mg / mL, 0.1 mg / mL, 0.2 mg / mL, 0.3 mg / mL, 0.4 mg / mL, 0.5 mg / mL, 0.6 mg / mL, 0.7 mg / mL, 0.8 mg / mL, 0.9 mg / mL, 1.0 mg / mL, 1.1 mg / mL, 1.2 mg / mL, 1.3 mg / mL, 1.4 mg / mL, 1.5 mg / mL, 1.6 mg / mL, 1.7 mg / mL, 1.8 mg / mL, 1.9 mg / mL, 2.0 mg / mL , 2.1mg / mL, 2.2mg / mL, 2.3mg / mL, 2.4mg / mL, 2.5mg / mL, 2.6mg / mL, 2.7mg / mL, 2.8mg / mL, 2.9mg / mL, 3.0mg / mL, 3.1mg / mL, 3.2mg / mL, 3.3mg / mL, 3.4mg / mL, 3.5mg / mL, 3.6 mg / mL, 3.7mg / mL, 3.8mg / mL, 3.9mg / mL, 4.0mg / mL, 4.1mg / mL, 4.2mg / mL, 4.3mg / mL, 4.4mg / mL, 4.5mg / mL, 4.6mg / mL, 4.7mg / mL, 4.8mg / mL, 4.9mg / mL, 5mg / ml or a value between any two data.

[0013] In some embodiments, the concentration of polysorbate 80 in the pharmaceutical composition is 0.08 to 0.5 mg / mL.

[0014] Further, the pH of the pharmaceutical composition described in the present disclosure is 3.0 to 8.0, including but not limited to 3.0, 4.0, 5.0, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0 or any value between two values.

[0015] In some embodiments, the pH of the pharmaceutical composition is between 6.0 and 7.0.

[0016] On the other hand, in some embodiments, the active ingredient in the pharmaceutical composition is present in the form of particles, and the particle size D90 of the active ingredient particles is less than 7 μm, selected from but not limited to 1.0 μm, 1.1 μm, 1.2 μm, 1.3 μm, 1.4 μm, 1.5 μm, 1.6 μm, 1.7 μm, 1.8 μm, 1.9 μm, 2.0 μm, 2.1 μm, 2.2 μm, 2.3 μm, 2.4 μm, 2.5 μm, 2.6 μm, 2.7μm, 2.8μm, 2.9μm, 3.0μm, 3.1μm, 3.2μm, 3.3μm, 3.4μm, 3.5μm, 3.6μm, 3.7μm, 3.8μm, 3.9 μm, 4.0μm, 4.1μm, 4.2μm, 4.3μm, 4.4μm, 4.5μm, 4.6μm, 4.7μm, 4.8μm, 4.9μm, 5.0μm or any value between two values.

[0017] In some embodiments, the particle size D90 of the active ingredient particles in the pharmaceutical composition is 2.0-4.0 μm.

[0018] In some embodiments, the particle size D90 of the active ingredient particles in the pharmaceutical composition is 2.0 to 3.5 μm.

[0019] In some embodiments, the particle size D90 of the active ingredient particles in the pharmaceutical composition is 3.0 μm or 3.2 μm.

[0020] In other embodiments, the particle size D50 of the active ingredient particles in the pharmaceutical composition is 1.0 μm, 1.1 μm, 1.2 μm, 1.3 μm, 1.4 μm, 1.5 μm, 1.6 μm, 1.7 μm, 1.8 μm, 1.9 μm, 2.0 μm, 2.1 μm, 2.2 μm, 2.3 μm, 2.4 μm, 2.5 μm, 2.6 μm, 2.7 μm, 2.8 μm, 2.9 μm, 3.0 μm, 3.1 μm, 3.2 μm, 3.3 μm, 3.4 μm, 3.5 μm, 3.6 μm, 3.7 μm, 3.8 μm, 3.9 μm, 4.0 μm or a value between any two values.

[0021] In some embodiments, the particle size D50 of the active ingredient particles in the pharmaceutical composition is 2.0-3.0 μm.

[0022] In some embodiments, the particle size D50 of the active ingredient particles in the pharmaceutical composition is 1.0-2.0 μm.

[0023] In some embodiments, the particle size D50 of the active ingredient particles in the pharmaceutical composition is 1.6 μm or 1.8 μm.

[0024] On the other hand, some embodiments provide pharmaceutical compositions in which the particle size D10 of the active ingredient particles is 0.1 μm, 0.2 μm, 0.3 μm, 0.4 μm, 0.5 μm, 0.6 μm, 0.7 μm, 0.8 μm, 0.9 μm or any value between two values.

[0025] On the other hand, the average particle size is expressed based on volume. Some embodiments provide a pharmaceutical composition in which the particle size D[4,3] of the active ingredient particles is 1.0 to 4 μm, including but not limited to 1.0 μm, 1.1 μm, 1.2 μm, 1.3 μm, 1.4 μm, 1.5 μm, 1.6 μm, 1.7 μm, 1.8 μm, 1.9 μm, 2.0 μm, 2.1 μm, 2.2 μm, 2.3 μm, 2.4 μm, 2.5 μm, 2.6 μm, 2.7 μm, 2.8 μm, 2.9 μm, 3.0 μm, 3.1 μm, 3.2 μm, 3.3 μm, 3.4 μm, 3.5 μm, 3.6 μm, 3.7 μm, 3.8 μm, 3.9 μm, 4.0 μm, or any value between two values.

[0026] In some embodiments, the particle size D[4,3] of the active ingredient particles in the pharmaceutical composition is 1.6 or 1.7 μm.

[0027] On the other hand, the pharmaceutical composition disclosed herein further contains sorbitan fatty acid ester.

[0028] In some embodiments, the concentration of sorbitan fatty acid esters in the pharmaceutical composition is 0.01 to 0.5 mg / mL, including but not limited to 0.01 mg / mL, 0.02 mg / mL, 0.04 mg / mL, 0.06 mg / mL, 0.08 mg / mL, 0.1 mg / mL, 0.2 mg / mL, 0.3 mg / mL, 0.4 mg / mL, 0.5 mg / mL or any value between two data.

[0029] On the other hand, the present disclosure also provides a pharmaceutical composition comprising particles of the active ingredient 9,10-dimethoxy-2-[[2-(2-oxo-imidazolin-1-yl)-ethyl]-(2,4,6-trimethyl-phenyl)-amino]-6,7-dihydro-pyrimidinyl[6,1-a]-isoquinolin-4-one or a pharmaceutically acceptable salt thereof, wherein the particle size D90 of the active ingredient particles is less than 7 μm.

[0030] In some embodiments, the particle size D90 of the active ingredient particles in the pharmaceutical composition is 2.0-4.0 μm, for example 3.0 μm or 3.2 μm.

[0031] In some embodiments, the particle size D50 of the active ingredient particles in the pharmaceutical composition is 1.6 μm or 1.8 μm.

[0032] In some embodiments, the particle size D[4,3] of the active ingredient particles in the pharmaceutical composition is 1.6 or 1.7 μm.

[0033] Furthermore, the aforementioned pharmaceutical composition also contains a tension regulator. In some embodiments, the tension regulator in the pharmaceutical composition is sodium chloride.

[0034] In some embodiments, the concentration of the tonicity modifier in the pharmaceutical composition is 2 to 8 mg / mL, including but not limited to 2.0 mg / mL, 2.2 mg / mL, 2.4 mg / mL, 2.6 mg / mL, 2.8 mg / mL, 3.0 mg / mL, 3.2 mg / mL, 3.4 mg / mL, 3.6 mg / mL, 3.8 mg / mL, 4.0 mg / mL, 4.2 mg / mL, 4.4 mg / mL, 4.6 mg / mL, 4.8 mg / mL, 5.0 mg / mL, 5.2 mg / mL, 5.4 mg / mL, 5.6 mg / mL, 5.8 mg / mL, 6.0 mg / mL, 6.2 mg / mL, 6.4 mg / mL, 6.6 mg / mL, 6.8 mg / mL, 7.0 mg / mL, 7.2 mg / mL, 7.4 mg / mL, 7.6 mg / mL, 7.8 mg / mL, 8.0 mg / mL or any value therebetween.

[0035] In some embodiments, the pharmaceutical composition contains 0.05 to 40 mg / mL of the active ingredient 9,10-dimethoxy-2-[[2-(2-oxo-imidazolin-1-yl)-ethyl]-(2,4,6-trimethyl-phenyl)-amino]-6,7-dihydro-pyrimidinyl[6,1-a]-isoquinolin-4-one or a pharmaceutically acceptable salt thereof, and 0.05 to 5.0 mg / mL of polysorbate 80.

[0036] In some embodiments, the pharmaceutical composition contains 0.05 to 40 mg / mL of the active ingredient 9,10-dimethoxy-2-[[2-(2-oxo-imidazolin-1-yl)-ethyl]-(2,4,6-trimethyl-phenyl)-amino]-6,7-dihydro-pyrimidinyl[6,1-a]-isoquinolin-4-one or a pharmaceutically acceptable salt thereof, and 0.05 to 5.0 mg / mL of polysorbate 80, wherein the active ingredient is present in particles.

[0037] In some embodiments, the pharmaceutical composition contains:

[0038] a) 0.05 to 40 mg / mL of the active ingredient 9,10-dimethoxy-2-[[2-(2-oxo-imidazolin-1-yl)-ethyl]-(2,4,6-trimethyl-phenyl)-amino]-6,7-dihydro-pyrimidinyl[6,1-a]-isoquinolin-4-one or a pharmaceutically acceptable salt thereof,

[0039] b) 0.05 to 5.0 mg / mL of polysorbate 80, and

[0040] c) 1 to 50 mM buffer.

[0041] In some embodiments, the pharmaceutical composition contains:

[0042] a) 0.05 to 40 mg / mL of the active ingredient 9,10-dimethoxy-2-[[2-(2-oxo-imidazolin-1-yl)-ethyl]-(2,4,6-trimethyl-phenyl)-amino]-6,7-dihydro-pyrimidinyl[6,1-a]-isoquinolin-4-one or a pharmaceutically acceptable salt thereof,

[0043] b) 0.05 to 5.0 mg / mL of polysorbate 80, and

[0044] c) 1 to 50 mM buffer, in which the active ingredient is present as particles.

[0045] In some embodiments, the pharmaceutical composition contains:

[0046] a) 0.05 to 40 mg / mL of the active ingredient 9,10-dimethoxy-2-[[2-(2-oxo-imidazolin-1-yl)-ethyl]-(2,4,6-trimethyl-phenyl)-amino]-6,7-dihydro-pyrimidinyl[6,1-a]-isoquinolin-4-one or a pharmaceutically acceptable salt thereof,

[0047] b) 0.05 to 5.0 mg / mL of polysorbate 80, and

[0048] c) 1 to 50 mM buffer, wherein the active ingredient is present in the form of particles having a particle size D90 of less than 7 μm.

[0049] In some embodiments, the pharmaceutical composition contains:

[0050] a) 0.05 to 40 mg / mL of the active ingredient 9,10-dimethoxy-2-[[2-(2-oxo-imidazolin-1-yl)-ethyl]-(2,4,6-trimethyl-phenyl)-amino]-6,7-dihydro-pyrimidinyl[6,1-a]-isoquinolin-4-one or a pharmaceutically acceptable salt thereof,

[0051] b) 0.05 to 5.0 mg / mL of polysorbate 80,

[0052] c) 1 to 50 mM buffer, and

[0053] d) 0.01 to 0.5 mg / mL of sorbitan fatty acid ester, wherein the active ingredient is present in the form of particles, and the particle size D90 of the particles is less than 7 μm.

[0054] Pharmaceutically acceptable salts of the active ingredients described in the present disclosure include, but are not limited to, fumarate, sulfate, and hydrochloride. In some embodiments, the pharmaceutically acceptable salt is a fumarate.

[0055] In some embodiments, the pharmaceutical compositions disclosed herein are suitable for administration via a nebulizer. For example, the aforementioned pharmaceutical compositions are delivered to the respiratory tract via a special device to exert local or systemic effects.

[0056] The present disclosure also provides a method for preparing the aforementioned pharmaceutical composition, comprising: grinding, wet-milling, homogenizing, precipitating or shearing 9,10-dimethoxy-2-[[2-(2-oxo-imidazolin-1-yl)-ethyl]-(2,4,6-trimethyl-phenyl)-amino]-6,7-dihydro-pyrimidinyl[6,1-a]-isoquinolin-4-one or a pharmaceutically acceptable salt thereof.

[0057] In some embodiments, for process and economic considerations, the active ingredient and all or part of the polysorbate 80 are generally ground together to prepare a concentrated suspension of the active ingredient, and then diluted to obtain a suitable dilute suspension. The remaining surface stabilizer can be mixed with the aforementioned composition during the dilution process to maintain the stability of the diluted liquid. On the other hand, in order to obtain D90 particles that meet the conditions in the embodiments of the present disclosure, particles of a suitable particle size are first obtained by micronization, and then ground to prepare a suspension after mixing with the aforementioned polysorbate 80.

[0058] In an optional embodiment, the aforementioned preparation method further comprises the step of mixing with a buffer and / or a tension regulator.

[0059] On the other hand, the pharmaceutical composition of the present disclosure can be further freeze-dried to obtain a solid composition, such as a freeze-dried powder injection. In some embodiments, the pharmaceutical composition is stored in a vial.

[0060] In some embodiments, the freeze-drying comprises the steps of pre-freezing, primary drying and secondary drying in sequence.

[0061] The present disclosure also provides a lyophilized composition obtained by freeze-drying the aforementioned pharmaceutical composition. Further, the lyophilized composition is reconstituted or re-dissolved in a liquid medium to obtain a pharmaceutical composition, and the liquid medium used for the reconstitution or re-dissolution is selected from water for injection.

[0062] The present disclosure also provides a pharmaceutical composition consisting of the following ingredients, a lyophilized composition prepared from the pharmaceutical composition, and a reconstituted solution obtained by further reconstitution or re-dissolution of the lyophilized composition with a liquid medium, wherein the pharmaceutical composition contains 9,10-dimethoxy-2-[[2-(2-oxo-imidazolin-1-yl)-ethyl]-(2,4,6-trimethyl-phenyl)-amino]-6,7-dihydro-pyrimidinyl[6,1-a]-isoquinolin-4-one or a pharmaceutically acceptable salt thereof.

[0063] On the other hand, the present disclosure also provides the use of the aforementioned pharmaceutical composition, lyophilized composition or reconstituted solution in the preparation of a medicament for preventing and / or treating asthma, obstructive pulmonary disease, sepsis, nephritis, diabetes, allergic rhinitis, allergic conjunctivitis, ulcerative enteritis or rheumatism, preferably obstructive pulmonary disease.

[0064] The present disclosure also provides a pharmaceutical composition for use in the prevention and / or treatment of asthma, obstructive pulmonary disease, sepsis, nephritis, diabetes, allergic rhinitis, allergic conjunctivitis, ulcerative colitis or rheumatism, preferably obstructive pulmonary disease.

[0065] the term

[0066] "Buffer" refers to a buffer that tolerates changes in pH through the action of its acid-base conjugate components. Examples of buffers that control pH in an appropriate range include, but are not limited to, citrate and phosphate.

[0067] "Phosphate buffer" or "phosphate buffer" is a buffer including phosphate ions. Examples of phosphate buffers include disodium hydrogen phosphate-sodium dihydrogen phosphate, disodium hydrogen phosphate-potassium dihydrogen phosphate, etc., and a preferred phosphate buffer is disodium hydrogen phosphate-sodium dihydrogen phosphate.

[0068] "Citrate buffer" is a buffer including acetate ions. Examples of citrate buffer include citric acid-sodium citrate and the like. A preferred citrate buffer is citric acid-sodium citrate buffer.

[0069] "Acetate buffer" is a buffer including acetate ions. Examples of acetate buffer include acetic acid-sodium acetate and the like. A preferred acetate buffer is acetic acid-sodium acetate buffer.

[0070] "Pharmaceutical composition" means a mixture containing one or more compounds described herein or their physiologically / pharmaceutically acceptable salts or prodrugs and other chemical components, such as physiologically / pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to maintain the stability of the active ingredient, promote administration to an organism, facilitate the absorption of the active ingredient and thus exert biological activity. Herein, "pharmaceutical composition" and "preparation" are not mutually exclusive.

[0071] "Lyophilized preparation" refers to a preparation or pharmaceutical composition obtained after a pharmaceutical composition or solution preparation in liquid or solution form has been subjected to a vacuum freeze-drying step.

[0072] The "D10" mentioned in the present disclosure refers to the particle size corresponding to when the cumulative particle size distribution percentage of a sample reaches 10%. "D50" refers to the particle size corresponding to when the cumulative particle size distribution percentage of a sample reaches 50%. "D90" refers to the particle size corresponding to when the cumulative particle size distribution percentage of a sample reaches 90%.

[0073] The "D[4,3]" mentioned in the present disclosure represents the average particle size based on volume. The full name is "mass-to-volume average particle size", which is abbreviated as volume average diameter. It is calculated by averaging the particle size values ​​at both ends of each particle size interval, multiplying them by the particle size distribution percentage corresponding to this interval, and then adding up the products, that is, D[4,3] = (f1·D1+f2·D2+f3·D3+……).

[0074] The content percentage (%) of all substances in the present disclosure refers to the weight-to-volume ratio. The "weight-to-volume ratio" mentioned in the present disclosure refers to the weight (in g) of the component contained in every 100 mL of liquid system, that is, g / 100 mL.

[0075] The numerical values ​​in this disclosure are instrumental measurements, which have a certain degree of error. Generally speaking, plus or minus 10% is within the reasonable error range. Of course, the context in which the numerical value is used needs to be considered. For example, the particle size of the active ingredient, the numerical value is the error change after measurement does not exceed plus or minus 10%, which can be plus or minus 9%, plus or minus 8%, plus or minus 7%, plus or minus 6%, plus or minus 5%, plus or minus 4%, plus or minus 3%, plus or minus 2% or plus or minus 1%, preferably plus or minus 5%.

[0076] Typical acceptable standards for stability of the present disclosure are as follows: the content change during storage is usually no more than about 10%, preferably no more than about 5% of the active ingredient is degraded as measured by HPLC. HPLC conditions: octadecylsilane bonded silica gel is used as the filler, phosphate, triethylamine phosphate buffer / acetonitrile is used as the mobile phase, and gradient elution is used.

[0077] Figure 1: Inhibitory effect on the number of immune cells in BALF (#p<0.01 vs normal control group, **p<0.01, *p<0.05 vs model control group).

[0078] Figure 2: Inhibitory effect on TNFα in BALF (**p<0.01, *p<0.05 vs model control group).

[0079] The following are specific implementation methods of the present disclosure. The examples are for further describing the present disclosure rather than limiting the present disclosure. All technical solutions equivalent to the present disclosure belong to the protection scope of the present disclosure.

[0080] Example 1

[0081] Preparation of 9,10-dimethoxy-2-[[2-(2-oxo-imidazolin-1-yl)-ethyl]-(2,4,6-trimethyl-phenyl)-amino]-6,7-dihydro-pyrimidinyl[6,1-a]isoquinolin-4-one (Compound 1) (see the method in WO2022228544)

[0082] Intermediate 1a: Preparation of 1-(2-chloroethyl)-imidazolin-2-one

[0083] At 0°C, thionyl chloride (5 mL) was slowly added to 1-(2-hydroxyethyl)imidazolidinone (3.5 g, 26.9 mmol), the temperature was raised to 45°C, and the mixture was stirred until the reaction was completed. A saturated sodium chloride solution was added to quench the reaction, and the pH was adjusted to 7 with a 10% NaOH solution. The mixture was extracted with dichloromethane, washed with a saturated sodium chloride solution, and dried over anhydrous sodium sulfate. The filtrate was concentrated under reduced pressure to obtain intermediate 1a (3.5 g, yield 88.4%), MS (ESI) m / z 149.1 [M+H] + .

[0084] Intermediate 1b: Preparation of 1-(3,4-dimethoxyphenethyl)urea

[0085] At room temperature, 2-(3,4-dimethoxyphenyl)ethylamine hydrochloride (4.3 g, 19.8 mmol) was dissolved in water (25 mL), the temperature was raised to 50°C, potassium cyanate (1.8 g, 21.8 mmol) was added in batches, stirring was continued until the reaction was completed, the temperature was lowered to 0°C, filtered, the filter cake was washed with ice water, and dried to obtain intermediate 1b (4.1 g, yield 93.8%), MS (ESI) m / z 225.1 [M+H] + .

[0086] Intermediate 1c: Preparation of 1-[2-(3,4-dimethoxy-phenyl)-ethyl]-pyrimidine-2,4,6-trione

[0087] Under ice bath condition, sodium ethoxide (3.8 g, 55.8 mmol) was added to anhydrous ethanol (50 mL) in batches. After the addition, the temperature was raised to reflux, and diethyl malonate (5.9 g, 36.6 mmol) was added dropwise. After the addition, stirring was continued for 0.25 h to 0.5 h. An ethanol solution (30 mL) of intermediate 1b (4.1 g, 18.3 mmol) was added dropwise. The reaction was stirred until the reaction was completed, and the temperature was lowered to 0°C. 5% HCl solution was added dropwise until pH = 6. 300 mL of water was added, and the mixture was filtered. The filter cake was washed with ice water and dried to obtain intermediate 1c (3.9 g, yield 77.1%), MS (ESI) m / z 293.1 [M+H] + .

[0088] Intermediate 1d: Preparation of 2-chloro-9,10-dimethoxy-6,7-dihydropyrimido[6,1-a]isoquinolin-4-one

[0089] At room temperature, intermediate 1c (3.9 g, 13.4 mmol) was added to phosphorus oxychloride (120 mL), the temperature was raised to 110°C, stirred until the reaction was completed, cooled and concentrated, the solid was poured into ice water, saturated NaOH solution was added dropwise until pH = 10, filtered, the filter cake was washed with ice water, and dried to obtain intermediate 1d (2.4 g, yield 62.4%), MS (ESI) m / z 293.1 [M+H] + .

[0090] Intermediate 1e: Preparation of 9,10-dimethoxy-2-(2,4,6-trimethylphenylimino)-2,3,6,7-tetrahydropyrimido[6,1-a]isoquinolin-4-one

[0091] At room temperature, intermediate 1d (2.4 g, 8.2 mmol) was suspended in isopropanol (30 mL), 2,4,6-trimethylaniline (4.5 g, 24.6 mmol) was added, the system was heated to 90 ° C, and stirring was continued until the reaction was completed, the temperature was lowered, and the filter cake was washed with ice water and dried to obtain intermediate 1e (3.0 g, yield 92.1%), MS (ESI) m / z 392.2 [M+H] + .

[0092] Compound 1: Preparation of 9,10-dimethoxy-2-[[2-(2-oxo-imidazolin-1-yl)-ethyl]-(2,4,6-trimethyl-phenyl)-amino]-6,7-dihydro-pyrimidinyl[6,1-a]isoquinolin-4-one

[0093] At room temperature, intermediate 1e (0.72 g, 1.8 mmol) was dissolved in tetrahydrofuran (20 mL), and potassium tert-butoxide (0.42 g, 3.6 mmol) was added under nitrogen atmosphere. After the addition, the temperature was raised to 65°C and stirring was continued for 48 h. The temperature was lowered to 25°C and intermediate 1a (0.82 g, 5.5 mmol) was added. After the addition, the temperature was raised to 80°C and stirring was continued until the reaction was completed. A saturated sodium chloride solution was added to quench the reaction, and the mixture was extracted with dichloromethane, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The target compound 1 (0.21 g, yield 46.5%) was obtained by silica gel column chromatography (n-heptane / ethyl acetate).

[0094] 1 H NMR (400MHz, CDCl3) δ6.99(s,2H),6.69(s,1H),6.64(s,1H),5.39(s,1H),4.61(s,1H),4.22-4.15(m,2H),4.08-3.99(m,2H),3.93 (s,3H),3.77-3.69(m,5H),3.55-3.46(m,2H),3.38-3.42(t,J=6.8Hz,2H),2.88-2.92(t,J=6.4Hz,2H),2.34(s,3H),2.18(s,6H).

[0095] MS (ESI) m / z 504.4 [M+H] + .

[0096] Test Example 1 In vitro PDE4B enzyme activity assay: IMAP FP-based assay

[0097] 1. Experimental Materials

[0098] 2. Experimental steps

[0099] The compound was serially diluted 5-fold with DMSO to obtain different concentrations (10000 nM, 2000 nM, 400 nM, 80 nM, 16 nM, 3.2 nM, 0.64 nM, 0.128 nM, 0.0256 nM, 0.005 nM). 200 μL of compounds of different concentrations were added to a 384-well plate (n=2), and 2 portions of 200 μL of DMSO were added to the 384-well plate (n=2) as a blank control; then 10 μL of 0.025 μg / mL PDE4B1 enzyme solution (prepared with 1 mM 5*IMAP Reaction buffer and 1 mM DTT) was added to the 384-well plate, and 10 μL of blank buffer without PDE4B1 enzyme was added to one of the blank controls, and incubated at room temperature for 15 minutes, and then 10 μL of 0.1 μM FAM-cAMP solution (prepared with 1 mM 5*IMAP Reaction buffer and 1 mM DTT) was added thereto, and after incubation at room temperature for 30 minutes, 60 μL of detection solution (prepared with 0.5625 mM 5*IMAP Progressive Binding buffer A, 0.1875 mM 5*IMAP Progressive Binding buffer B and 0.75mM beads), incubated at room temperature for 60 minutes and collected data. The inhibition rate calculation formula is: Inhibition rate = M / (MM control )*100; IC was calculated based on the concentration and inhibition rate fitting curve 50 In this experiment, RPL554 was used as a positive control.

[0100] The examples disclosed herein inhibited PDE4B1 enzyme activity in vitro by the above test, and the measured IC 50 =50nM.

[0101] Test Example 2 In vitro PDE3A enzyme activity assay: IMAP FP-based assay

[0102] 1. Experimental Materials

[0103] 2. Experimental steps

[0104] The compound was serially diluted 5-fold with DMSO to obtain different concentrations (10000 nM, 2000 nM, 400 nM, 80 nM, 16 nM, 3.2 nM, 0.64 nM, 0.128 nM, 0.0256 nM, 0.005 nM). 200 μL of compounds of different concentrations were added to a 384-well plate (n=2), and 2 portions of 200 μL of DMSO were added to the 384-well plate (n=2) as a blank control; then 10 μL of 0.025 μg / mL PDE4B1 enzyme solution (prepared with 1 mM 5*IMAP Reaction buffer and 1 mM DTT) was added to the 384-well plate, and 10 μL of blank buffer without PDE3A enzyme was added to one of the blank controls, and incubated at room temperature for 15 minutes, and then 10 μL of 0.1 μM FAM-cAMP solution (prepared with 1 mM 5*IMAP Reaction buffer and 1 mM DTT) was added thereto, and after incubation at room temperature for 30 minutes, 60 μL of detection solution (prepared with 0.5625 mM 5*IMAP Progressive Binding buffer A, 0.1875 mM 5*IMAP Progressive Binding buffer B and 0.75mM beads), incubated at room temperature for 60 minutes and collected data. The inhibition rate calculation formula is: Inhibition rate = M / (MM control )*100; IC was calculated based on the concentration and inhibition rate fitting curve 50 In this experiment, RPL554 was used as a positive control.

[0105] The examples disclosed herein inhibited PDE3A enzyme activity in vitro by the above test, and the measured IC 50 =0.13nM.

[0106] Example 2

[0107] 9,10-dimethoxy-2-[[2-(2-oxo-imidazolin-1-yl)-ethyl]-(2,4,6-trimethyl-phenyl)-amino]-6,7-dihydro-pyrimidinyl[6,1-a]-isoquinolin-4-one (Compound A) and the excipients in Table 1 were prepared according to the following steps to prepare different formulations.

[0108] Step 1: Prepare buffer solution: Weigh the prescribed amount of polysorbate 20, Span 20, polysorbate 80, sodium dihydrogen phosphate, sodium hydrogen phosphate and sodium chloride respectively, and stir to dissolve with appropriate amount of water.

[0109] Step 2: Add the prescribed amount of compound A, dilute to 1 / 20 of the total preparation volume with the buffer obtained in step 1, and stir evenly; shear with a high-speed shearing machine T50 (shearing speed of about 5000 rpm) for 2-6 minutes, sterilize, and obtain a concentrated suspension.

[0110] Step 3: Add the buffer obtained in step 1 to the concentrated suspension in step 2, mix, adjust the volume, and shear with a high-speed shearing machine T50 for 10-30 minutes.

[0111] Fill, pack into polyester / aluminum / polyethylene pharmaceutical composite bags, and store in sealed containers.

[0112] Table 1: Prescription quantity configuration

[0113] The above prescriptions 1-5 were placed under different conditions to investigate their stability. The data are shown in Table 2

[0114] Note: d stands for day and M stands for month.

[0115] Conclusion: The Chinese Pharmacopoeia Part IV General Rules for Inhalation Preparations stipulates that the particle size of drugs in inhalation preparations should generally be controlled below 10 μm, and most of them should be 5 μm. Under long-term storage conditions or accelerated storage conditions, the particle size of the active ingredients in the composition of Prescriptions 1 and 2 increased significantly, such as the D90 value greater than 5 μm, or even greater than 10 μm. Compared with polysorbate 20, the formulation containing polysorbate 80 can maintain a stable particle size under accelerated conditions or long-term storage at 25°C, showing better composition storage stability.

[0116] The prescription 5 FPF (fine particle fraction) was measured to be 52.64% using a breathing simulator (model: Copley BRS2100) and a nebulizer pump (model: PARI TurboBOY) to simulate adult breathing (simulation parameters: tidal volume 500 ml; respiratory rate 15 cycles / min; sinusoidal respiratory waveform; respiratory ratio 1:1).

[0117] Example 3

[0118] 9,10-dimethoxy-2-[[2-(2-oxo-imidazolin-1-yl)-ethyl]-(2,4,6-trimethyl-phenyl)-amino]-6,7-dihydro-pyrimidinyl[6,1-a]-isoquinolin-4-one (Compound A) and the excipients in Table 3 were prepared according to the steps of Example 2 to prepare different formulations.

[0119] Table 3: Prescription quantity configuration

[0120] The above prescriptions 6-9 were placed under different conditions to investigate their stability. The data are shown in Table 4.

[0121] Table 4

[0122] Example 4

[0123] Step 1: Prepare buffer solution: weigh the prescribed amount of polysorbate 80, sodium dihydrogen phosphate, disodium hydrogen phosphate and sodium chloride, and stir to dissolve with appropriate amount of water.

[0124] Step 2: Add the prescribed amount of compound A, dilute to 1 / 20 of the total preparation volume with the buffer obtained in step 1, and stir evenly; shear with a high-speed shearing machine T50 (shearing speed of about 5000 rpm) for 2-6 minutes, sterilize, and obtain a concentrated suspension.

[0125] Step 3: Add the buffer obtained in step 1 to the concentrated suspension in step 2, mix, adjust the volume, and shear with a high-speed shearing machine T50 for 10-30 minutes.

[0126] Table 5

[0127] Note: The batch size is 20,000 pieces

[0128] The above formulation was placed at 40°C ± 2°C / 25% RH ± 5% RH to examine its stability. The data are shown in Table 6.

[0129] Table 6

[0130] Example 5: Drug efficacy study in LPS-induced acute lung inflammation model in mice

[0131] This study used the LPS-induced acute lung inflammation model of BALB / c mice, and evaluated the inhibitory effect of the compound on lung inflammation by counting immune cells and the level of TNFα cytokines in bronchoalveolar lavage fluid (BALF). The mice were divided into 6 groups, 8 in each group, including: normal control group, model control group, RPL554 group (1.0 mg / kg), low, medium and high dose groups of compound A (0.3, 1.0, 3.0 mg / kg). Mice were stimulated by LPS aerosolization 1 hour after intratracheal aerosol administration, and the BALF of the animals was collected 6 hours after modeling, and the total cells, neutrophils, and eosinophils in the BALF were counted, and the level of proinflammatory cytokine TNFα in the BALF was detected.

[0132] The results of cell counting in BALF are shown in Figure 1. The number of total cells, neutrophils and eosinophils in the BALF of the model control group animals was significantly higher than that of the normal control group. After administration of 1.0 mg / kg to the RPL554 positive control group, the number of total cells, neutrophils and eosinophils in the BALF was significantly reduced compared with the model group. The number of neutrophils and eosinophils in the compound A 0.3, 1.0 and 3.0 mg / kg groups were also significantly reduced compared with the model group, and the efficacy of compound A 0.3 mg / kg was equivalent to that of RPL554 1 mg / kg; the 1.0 and 3.0 mg / kg groups showed a significant decrease in the number of total cells, and the 0.3 mg / kg group showed a decreasing trend.

[0133] The results of TNFα level detection in BALF are shown in Figure 2. The level of TNFα in BALF of the model control group was significantly higher than that of the normal control group. After administration of 1.0 mg / kg to the RPL554 positive control group, the level of TNFα in BALF showed a downward trend compared with the model group. All dose groups of compound A can reduce the level of TNFα, among which the 0.3 and 1.0 mg / kg groups were significantly lower than those of the model group.

[0134] Example 6

[0135] 9,10-dimethoxy-2-[[2-(2-oxo-imidazolin-1-yl)-ethyl]-(2,4,6-trimethyl-phenyl)-amino]-6,7-dihydro-pyrimidinyl[6,1-a]-isoquinolin-4-one (Compound A), sodium carboxymethyl cellulose, polysorbate 20, sodium dihydrogen phosphate, sodium dihydrogen phosphate and sodium chloride were weighed according to the amounts in Table 7, and a formulation composition was prepared according to the following steps.

[0136] Table 7

[0137] Step 1: Prepare buffer solution: Weigh the prescribed amount of polysorbate 20, sodium dihydrogen phosphate, disodium hydrogen phosphate and sodium chloride, and stir to dissolve with appropriate amount of water.

[0138] Step 2: Add the prescribed amount of compound A, dilute to 1 / 20 of the total preparation volume with the buffer obtained in step 1, and stir evenly; shear with a high-speed shearing machine T50 (shearing speed of about 5000 rpm) for 2-6 minutes, sterilize, and obtain a concentrated suspension.

[0139] Step 3: Add the buffer obtained in step 1 to the concentrated suspension in step 2, mix, adjust the volume, and shear with a high-speed shearing machine T50 for 10-30 minutes.

[0140] The above prescriptions 12-14 were placed under different conditions to investigate their stability. The data are shown in Table 10.

[0141] Table 8

[0142] Note: / The data of this batch fluctuates greatly and is inaccurate.

[0143] The FPF of prescription 12 was measured to be 44.1% using a respiratory simulator (model: Copley BRS2100) and a nebulizer pump (model: PARI TurboBOY) to simulate adult breathing (simulation parameters: tidal volume 500 ml; respiratory rate 15 cycles / min; sinusoidal respiratory waveform; respiratory ratio 1:1).

Claims

A pharmaceutical composition comprises active ingredients 9,10-dimethoxy-2-[[2-(2-oxo-imidazolin-1-yl)-ethyl]-(2,4,6-trimethyl-phenyl)-amino]-6,7-dihydro-pyrimidinyl[6,1-a]-isoquinolin-4-one or a pharmaceutically acceptable salt thereof, and polysorbate 80. The pharmaceutical composition according to claim 1, wherein the active ingredient concentration is 0.01 to 40 mg / mL, preferably 0.05 to 20 mg / mL. The pharmaceutical composition according to claim 1 or 2, further comprising a buffer, wherein the buffer is preferably a citrate buffer or a phosphate buffer or an acetate buffer, more preferably a phosphate buffer, and most preferably a sodium dihydrogen phosphate-disodium hydrogen phosphate buffer. The pharmaceutical composition according to any one of claims 1 to 3, wherein the concentration of polysorbate 80 is 0.05 to 5.0 mg / mL, preferably 0.08 to 0.5 mg / mL. The pharmaceutical composition according to any one of claims 1 to 4, wherein the concentration of the buffer is 1 to 50 mM, preferably 5 to 30 mM. The pharmaceutical composition according to any one of claims 1 to 5, wherein the pH of the composition is 3.0 to 8.

0. The pharmaceutical composition according to any one of claims 1 to 6, further comprising sorbitan fatty acid ester. The pharmaceutical composition according to claims 1-7, wherein the active ingredient is present in the form of particles, the particle size D90 of the particles is less than 7 μm, preferably 1.0 to 5.0 μm, for example 3.0 μm or 3.2 μm. A pharmaceutical composition comprising: a) 0.05 to 40 mg / mL of an active ingredient 9,10-dimethoxy-2-[[2-(2-oxo-imidazolin-1-yl)-ethyl]-(2,4,6-trimethyl-phenyl)-amino]-6,7-dihydro-pyrimidinyl[6,1-a]-isoquinolin-4-one or a pharmaceutically acceptable salt thereof, b) 0.05 to 2.0 mg / mL of polysorbate 80, and c) 1 to 50 mM of a buffer. A pharmaceutical composition comprising particles of an active ingredient 9,10-dimethoxy-2-[[2-(2-oxo-imidazolin-1-yl)-ethyl]-(2,4,6-trimethyl-phenyl)-amino]-6,7-dihydro-pyrimidinyl[6,1-a]-isoquinolin-4-one or a pharmaceutically acceptable salt thereof, wherein the particle size D90 of the active ingredient particles is less than 7 μm, preferably 1.0 to 5.0 μm, for example 3.0 μm or 3.2 μm The pharmaceutical composition according to any one of claims 1 to 10, further comprising a tension regulator, wherein the tension regulator is preferably sodium chloride. The pharmaceutical composition according to any one of claims 1 to 11, which is suitable for administration via a nebulizer. A method for preparing the pharmaceutical composition according to any one of claims 1 to 12, comprising: The active ingredient 9,10-dimethoxy-2-[[2-(2-oxo-imidazolin-1-yl)-ethyl]-(2,4,6-trimethyl-phenyl)-amino]-6,7-dihydro-pyrimidinyl[6,1-a]-isoquinolin-4-one or a pharmaceutically acceptable salt thereof is subjected to a grinding, wet-milling, homogenization, precipitation or shearing step. A method for preparing a lyophilized preparation containing 9,10-dimethoxy-2-[[2-(2-oxo-imidazolin-1-yl)-ethyl]-(2,4,6-trimethyl-phenyl)-amino]-6,7-dihydro-pyrimidinyl[6,1-a]-isoquinolin-4-one or a pharmaceutically acceptable salt thereof, comprising the step of freeze-drying the pharmaceutical composition according to any one of claims 1 to 12. A lyophilized preparation containing 9,10-dimethoxy-2-[[2-(2-oxo-imidazolin-1-yl)-ethyl]-(2,4,6-trimethyl-phenyl)-amino]-6,7-dihydro-pyrimidinyl[6,1-a]-isoquinolin-4-one or a pharmaceutically acceptable salt thereof prepared by the method of claim 14. A method for preparing a reconstituted solution containing 9,10-dimethoxy-2-[[2-(2-oxo-imidazolin-1-yl)-ethyl]-(2,4,6-trimethyl-phenyl)-amino]-6,7-dihydro-pyrimidinyl[6,1-a]-isoquinolin-4-one or a pharmaceutically acceptable salt thereof, comprising the step of reconstituted the lyophilized preparation according to claim 15, wherein the liquid medium used for reconstitution is preferably water for injection. A reconstituted solution containing 9,10-dimethoxy-2-[[2-(2-oxo-imidazolin-1-yl)-ethyl]-(2,4,6-trimethyl-phenyl)-amino]-6,7-dihydro-pyrimidinyl[6,1-a]-isoquinolin-4-one or a pharmaceutically acceptable salt thereof prepared by the method of claim 16. A nebulizer comprising the pharmaceutical composition according to any one of claims 1 to 12, the lyophilized preparation according to claim 15, or the reconstituted solution according to claim 17. Use of the pharmaceutical composition according to any one of claims 1 to 12, the lyophilized preparation according to claim 15 or the reconstituted solution according to claim 17 in the preparation of a medicament for preventing and / or treating asthma, obstructive pulmonary disease, sepsis, nephritis, diabetes, allergic rhinitis, allergic conjunctivitis, ulcerative enteritis or rheumatism, preferably obstructive pulmonary disease.