Crystal form of pde3 / 4 dual inhibitor, and preparation method therefor and use thereof

Abstract

Provided in the present invention are a crystal form of a compound of formula (I), a preparation method therefor, and the use thereof in the preparation of a related drug.

Description

A crystalline form of a PDE3 / 4 dual inhibitor, its preparation method and application Technical Field

[0001] This invention relates to the polymorphism of a compound, its preparation method and application, specifically to the multiple polymorphisms of a PDE3 / 4 dual inhibitor, its preparation method and application, belonging to the field of medicinal chemistry technology. Background Technology

[0002] COPD (chronic obstructive pulmonary disease) is a group of lung diseases characterized by airflow limitation that is not fully reversible and progresses, primarily affecting the lungs. It is the most common chronic killer of lung health. The high incidence and mortality rates of COPD are due to two main factors: firstly, early diagnosis is extremely difficult, as COPD often has an insidious onset, and once clinical manifestations appear, it often indicates a gradual decline in the patient's overall health and a gradual increase in respiratory symptoms; secondly, COPD is currently incurable, and medications primarily rely on bronchodilators that modulate airway smooth muscle. These drugs only relieve symptoms and delay disease progression, treating the symptoms but not the root cause. COPD has a long course, often requiring frequent medical visits, hospitalizations for acute exacerbations, and long-term care, consuming significant medical resources and becoming a serious global disease burden. Therefore, for COPD, it is necessary to develop new drugs targeting different areas to meet diverse clinical needs.

[0003] Phosphodiesterases (PDEs) belong to a superfamily of enzymes, comprising 11 families, each involved in different signal transductions and regulating various physiological processes. Studies have found that PDE3 is associated with the contraction of respiratory smooth muscle, while PDE4 plays a crucial role in inflammatory responses induced by immune cells. Given the limitations of selective PDE3 or PDE4 inhibitors in clinical practice, dual PDE3 / 4 inhibitors appear to be a more attractive approach for targeting key pathological features of COPD and asthma. Current evidence suggests that dual PDE3 / 4 inhibitors have synergistic inhibitory effects, including synergistic anti-inflammatory and bronchodilatory effects. WO2000058308A1 reported that compound RPL554 has long-acting bronchodilatory and anti-inflammatory effects; however, this drug has poor solubility and high plasma clearance, making it suitable for inhalation administration. Bioactivity data show that its PDE4 inhibitory activity is unsatisfactory, potentially leading to less than ideal anti-inflammatory effects. Therefore, dual PDE3 / 4 inhibitors warrant further investigation.

[0004] WO2023109802 discloses a compound of formula (I) which exhibits excellent PDE3 and PDE4 inhibitory activity.

[0005] This invention describes multiple crystal forms of a compound of formula (I), which belongs to the PDE3 / 4 dual inhibitors and is intended for the treatment or prevention of PDE3 / 4 mediated diseases. The preparation method of the compound is also disclosed. Summary of the Invention

[0006] This invention provides a crystal form of the compound of formula (I) that is high in purity, high in stability, high in bioavailability, low in hygroscopicity, and has a uniform particle size distribution;

[0007] This invention provides a crystalline form of a compound of formula (I).

[0008] The present invention provides crystal form 1 of the compound of formula (I) above, which, in some embodiments, is irradiated with Cu-Kα and has X-ray powder diffraction patterns with characteristic diffraction peaks at the following 2θ positions: 10.35°±0.2°, 13.12°±0.2°, 15.56°±0.2°, 17.95°±0.2°, and 21.34°±0.2°.

[0009] In some embodiments, the crystal form 1 of the aforementioned compound of formula (I), when irradiated with Cu-Kα, has a characteristic diffraction peak at the following 2θ positions in its X-ray powder diffraction pattern: 10.35°±0.2°, 12.83°±0.2°, 13.12°±0.2°, 14.99°±0.2°, 15.56°±0.2°, 17.95°±0.2°, 21.34°±0.2°, and 22.81°±0.2°.

[0010] In some embodiments, the crystal form 1 of the aforementioned compound of formula (I), when irradiated with Cu-Kα, has a characteristic diffraction peak at the following 2θ positions in its X-ray powder diffraction pattern: 6.54°±0.2°, 10.35°±0.2°, 12.83°±0.2°, 13.12°±0.2°, 14.47°±0.2°, 14.99°±0.2°, 15.56°±0.2°, 16.12°±0.2°, 17.95°±0.2°, 21.34°±0.2°, 22.81°±0.2°, 23.25°±0.2°, 29.18°±0.2°, and 31.53°±0.2°.

[0011] In one embodiment, the crystal form 1 of the aforementioned compound of formula (I) is subjected to Cu-Kα radiation, and its X-ray powder diffraction pattern is shown in Figure 1.

[0012] In one embodiment, the crystal form 1 of the aforementioned compound of formula (I) has a peak temperature of 249.88°C as shown by differential scanning calorimetry (DSC) and a weight loss of approximately 0.37% before 236°C as shown by thermogravimetric analysis (TGA). The differential scanning calorimetry and thermogravimetric analysis curves are shown in Figures 2 and 3, respectively.

[0013] In one embodiment, the hygroscopicity (DVS) isotherm of the aforementioned compound of formula (I) shows that the water adsorption is 0.1402% in the range of 0RH%-80%RH, and its isotherm adsorption curve is shown in Figure 4.

[0014] In some embodiments, the crystal structure of compound crystal form 1 shown in formula (I) was determined by single-crystal diffraction. The unit cell parameters are shown in Table 1. This crystal structure belongs to the triclinic crystal system and has no helical axis. Space group, cell constant is α = 91.511(2)°, γ = 90.364(2)°, β = 99.668(2)°, cell volume The Z' of the system is 2; in some embodiments, the asymmetric unit of crystal form 1 consists of two compound molecules as shown in formula (I); in some embodiments, the asymmetric unit in the unit cell is as shown in Figure 5.

[0015] Table 1 shows the unit cell parameters of crystal form 1 of the compound represented by formula (I):

[0016] The present invention also provides an amorphous form of the compound of formula (I) above, and its X-ray powder diffraction pattern using Cu-Kα radiation is shown in Figure 6.

[0017] In one embodiment, the amorphous form of the aforementioned compound (I) has a Tg temperature of 95.70°C as shown by differential scanning calorimetry (mDSC) and a weight loss of approximately 2.42% before 150°C as shown by thermogravimetric analysis (TGA). The differential scanning calorimetry and thermogravimetric analysis curves are shown in Figures 7 and 8, respectively.

[0018] In one embodiment, the amorphous form of the aforementioned compound (I) has a hygroscopicity (DVS) isotherm curve showing that the water adsorption at 0RH%-80%RH is 3.865%, which is significantly higher than that of crystal form 1. Its isotherm adsorption curve is shown in Figure 9.

[0019] The present invention also provides a method for preparing crystal form 1 of the compound of formula (I), wherein the method comprises: using the compound of formula (I) as a raw material to perform slurry crystallization, evaporation crystallization, solvent-resistant crystallization, cooling crystallization, water vapor stress crystallization, and diffusion crystallization; in some embodiments, the solvent used is selected from C1-6 Halogenated alkane solvents, C 2-6 Ester solvents, C 2-6 Nitrile solvents, C 2-6 Ether solvents, C 1-6 One or more of an alcohol solvent or a mixture of solvents in any proportion, preferably one or more of dichloromethane, 1,2-dichloroethane, acetonitrile, ethyl acetate, methanol, ethanol, isopropanol, propanol, diethyl ether, tetrahydrofuran, or water.

[0020] The crystalline and amorphous forms of the compounds of formula (I) of this invention have advantages including, but not limited to, ease of processing and crystallization, ease of handling, ease of purification, ease of industrialization, good flowability, ease of micronization, high solubility, good pharmacokinetic characteristics and good stability, making them suitable for the preparation of pharmaceutical formulations.

[0021] The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of the aforementioned crystalline form, and a pharmaceutically acceptable carrier and / or excipient.

[0022] The present invention also provides the use of the crystal form or pharmaceutical composition described in any of the foregoing embodiments in the preparation of medicaments for treating or preventing PDE3 / 4 mediated diseases.

[0023] The present invention also provides a method for treating or preventing PDE3 / 4-mediated diseases, the method comprising administering a subject a therapeutically effective amount of the crystal form or combination thereof described in any of the foregoing embodiments.

[0024] The crystalline form of the compound of formula (I) described in this invention possesses excellent physical properties, including but not limited to solubility, dissolution rate, light resistance, low hygroscopicity, high temperature resistance, and high humidity resistance. For example, the crystalline form described in this invention can significantly reduce filtration time, shorten the production cycle, and save costs during formulation. The crystalline form described in this invention also has good photostability, thermal stability, and moisture stability, ensuring the reliability of the crystalline form during storage and transportation, thereby guaranteeing the safety of the formulation. Furthermore, the crystalline form does not require special packaging to prevent the effects of light, temperature, and humidity, thus reducing costs. The crystalline form will not degrade due to light, high temperature, and high humidity, improving the safety of the formulation and its effectiveness after long-term storage. Patients taking the crystalline form will not worry about photosensitivity reactions caused by exposure to sunlight.

[0025] The crystal form of the compound of formula (I) described in this invention exhibits good chemical and physical stability, is easy to prepare, and is more suitable for formulation preparation. The crystal form of this invention has good flowability, good compressibility, high bulk density, low hygroscopicity, and uniform particle size distribution.

[0026] The crystal form of the compound of formula (I) described in this invention is suitable and convenient for large-scale preparation. The preparation obtained by using the aforementioned crystal form can reduce irritation and improve absorption, thereby solving the problem of metabolic rate, significantly reducing toxicity, improving safety, and effectively ensuring the quality and efficacy of the preparation.

[0027] It can be understood that expressions such as "preferably, ..., its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ positions", or "more preferably, ..., its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ positions", etc., in this invention refer to the presence of characteristic diffraction peaks at the aforementioned 2θ positions, in addition to the presence of characteristic diffraction peaks at the aforementioned 2θ positions, also including characteristic diffraction peaks at the aforementioned "the following 2θ positions".

[0028] It is understood that the numerical values ​​described and protected in this invention are approximate. Variations within these values ​​may be attributed to equipment calibration, equipment errors, crystal purity, crystal size, sample size, and other factors.

[0029] The crystal structure of the present invention can be analyzed using various analytical techniques known to those skilled in the art, including but not limited to X-ray powder diffraction (XRD), ion chromatography (IC), differential scanning calorimetry (DSC), and / or thermogravimetric analysis (TGA), also known as thermogravimetry (TG).

[0030] It is understood that the crystal forms of the present invention are not limited to those that are exactly the same as the characteristic spectra described in the accompanying drawings, such as XRD, DSC, and TGA. Any crystal form that has a characteristic spectra that are substantially the same or essentially the same as those described in the accompanying drawings falls within the scope of the present invention.

[0031] It is understood that, as is well known in the field of differential scanning calorimetry (DSC), the melting peak height of a DSC curve depends on many factors related to sample preparation and instrument geometry, while the peak position is relatively insensitive to experimental details. Therefore, in some embodiments, the crystalline compounds of the present invention are characterized by DSC plots with characteristic peak positions, possessing substantially the same properties as the DSC plots provided in the accompanying drawings of the present invention, with an error tolerance of ±3°C.

[0032] Unless otherwise stated, the technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. In case of any conflict, the definitions provided herein shall prevail. When a quantity, concentration, or other value or parameter is expressed as a range, preferred range, or preferred upper and lower numerical limits, it should be understood that this is equivalent to specifically disclosing any range by combining any pair of upper or preferred values ​​with any lower or preferred value, regardless of whether the range is specifically disclosed. Unless otherwise stated, the numerical ranges listed herein are intended to include the endpoints of the range and all integers and fractions (decimals) within that range.

[0033] Unless otherwise stated, the terms used in the specification and claims have the following meanings.

[0034] The terms "optional" or "optionally" used in this invention refer to events or circumstances that may, but are not required to, occur as described below, including situations in which the event or circumstance may or may not occur.

[0035] When used in conjunction with a numerical variable, the terms "about" or "approximately" in this invention typically refer to the value of the variable and all values ​​of the variable being within the experimental error range (e.g., within a 95% confidence interval for the average value) or within ±10% of the specified value, or a wider range.

[0036] Unless otherwise stated, all percentages, parts, etc. in this document are by weight.

[0037] The “crystal form” or “crystal” mentioned in this invention refers to any solid substance exhibiting a three-dimensional arrangement, which, in contrast to amorphous solid substances, produces a characteristic XRPD pattern with clearly defined peaks.

[0038] The “pharmaceutical composition” described in this invention refers to a mixture of one or more compounds described herein or their physiologically / pharmaceutically acceptable salts with other components, wherein the other components include physiologically / pharmaceutically acceptable carriers and excipients.

[0039] The "carrier" as used in this invention refers to a carrier or diluent that does not cause significant irritation to organisms and does not eliminate the biological activity and properties of the given compound.

[0040] The term "excipient" as used in this invention refers to an inert substance added to a pharmaceutical composition for compound-dependent administration. Examples of excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars and different types of starch, cellulose derivatives (including microcrystalline cellulose), gelatin, vegetable oils, polyethylene glycols, diluents, granulating agents, lubricants, binders, disintegrants, etc.

[0041] The "ether solvents" mentioned in this invention refer to chain or cyclic compounds containing an ether bond -O- and having 1 to 10 carbon atoms. Specific examples include, but are not limited to, tetrahydrofuran, diethyl ether, propylene glycol methyl ether, methyl tert-butyl ether, isopropyl ether, or 1,4-dioxane.

[0042] The "alcohol solvent" mentioned in this invention refers to one or more "hydroxyl groups" replacing "C". 1-6 A group derived from one or more hydrogen atoms on an alkyl group, wherein the hydroxyl group and the C 1-6 "Alkyl" as defined above, specific examples include but are not limited to: methanol, ethanol, isopropanol, n-propanol, isoamyl alcohol, or trifluoroethanol.

[0043] The "ester solvent" mentioned in this invention refers to a combination of a lower organic acid containing 1 to 4 carbon atoms and a lower alcohol containing 1 to 6 carbon atoms. Specific examples include, but are not limited to, ethyl acetate, isopropyl acetate, or butyl acetate.

[0044] The "ketone solvents" mentioned in this invention refer to compounds in which a carbonyl group (-C(O)-) is attached to two hydrocarbon groups. Depending on the hydrocarbon groups in the molecule, ketones can be classified into aliphatic ketones, alicyclic ketones, aromatic ketones, saturated ketones, and unsaturated ketones. Specific examples include, but are not limited to, acetone, acetophenone, and 4-methyl-2-pentanone.

[0045] The "nitrile solvent" mentioned in this invention refers to one or more "cyano" groups replacing "C". 1-6 A group derived from one or more hydrogen atoms on an alkyl group, wherein the "cyano" and "C" 1-6 "Alkyl" is as defined above, and specific examples include, but are not limited to, acetonitrile or propionitrile.

[0046] The "halogenated hydrocarbon solvent" described in this invention refers to one or more "halogen atoms" replacing "C". 1-6 A group derived from one or more hydrogen atoms on an alkyl group, wherein the halogen atom and the C 1-6 "Alkyl" is defined above, and specific examples include, but are not limited to, dichloromethane, 1,2-dichloroethane, chloroform, or carbon tetrachloride.

[0047] The "room temperature" mentioned in this invention generally refers to 4 to 30°C, and preferably to 20±5°C.

[0048] The drying temperature described in this invention is generally 20–100°C, preferably 25–70°C. It can be performed under normal pressure or under reduced pressure (vacuum drying). Preferably, the drying is carried out under reduced pressure.

[0049] The "X-ray powder diffraction pattern (XRPD pattern)" mentioned in this invention refers to an experimentally observed diffraction pattern or parameters, data, or values ​​derived from it. XRPD patterns are typically characterized by peak position (horizontal axis) and / or peak intensity (vertical axis).

[0050] The "2θ or 2θ angle" mentioned in this invention refers to the diffraction angle, where θ is the Bragg angle, which is the peak position expressed in degrees (°) based on the setup in an X-ray diffraction experiment, and is usually the horizontal axis unit in the diffraction pattern. If the incident beam is diffracted when it forms an angle θ with a certain lattice plane, the experimental setup needs to record the reflected beam at a 2θ angle. It should be understood that the specific 2θ value for a particular crystal form mentioned herein is intended to represent the 2θ value (expressed in degrees) measured using the X-ray diffraction experimental conditions described herein, with an error range of ±0.3, which can be ±0.3, ±0.2, or ±0.1.

[0051] The term "substantially identical" as used in this invention means taking into account representative peak positions and intensity variations. For example, those skilled in the art will understand that peak positions (2θ) will exhibit some variation, typically up to 0.1–0.2 degrees, and that the instrument used to measure diffraction will also cause some variation. Furthermore, those skilled in the art will understand that relative peak intensities will vary due to differences between instruments, as well as the degree of crystallinity, preferred orientation, the surface of the prepared sample, and other factors known to those skilled in the art, and should be considered as qualitative measurements only.

[0052] The "differential scanning calorimetry or DSC" described in this invention refers to measuring the temperature difference and heat flow difference between the sample and the reference material during the sample heating or isothermal process, in order to characterize all physical and chemical changes related to thermal effects and obtain phase transition information of the sample.

[0053] According to the description of hygroscopic characteristics and the definition of hygroscopic weight gain in the "Guiding Principles on Hygroscopicity of Drugs" in Part IV of the 2020 edition of the Chinese Pharmacopoeia:

[0054] Deliquescence: Absorbs sufficient moisture to form a liquid;

[0055] Extremely hygroscopic: the weight gain due to hygroscopic absorption is not less than 15%;

[0056] It has hygroscopic properties: the weight gain due to hygroscopic absorption is less than 15% but not less than 2%;

[0057] Slightly hygroscopic: the weight gain due to moisture absorption is less than 2% but not less than 0.2%;

[0058] It has little or no hygroscopicity: the weight gain due to moisture absorption is less than 0.2%.

[0059] The crystallization methods described in this invention include, but are not limited to, slurry crystallization, volatile crystallization, antisolvent crystallization, cooling crystallization, water vapor stress crystallization, or diffusion crystallization.

[0060] The crystal form disclosed in this invention can be prepared by the following method:

[0061] 1. The evaporation experiment involves evaporating a clear solution of the sample in an open container at different temperatures until the solvent is dry.

[0062] 2. The crystal slurry experiment involves stirring a supersaturated solution of the sample (containing insoluble solids) in different solvent systems at a certain temperature.

[0063] 3. The solvent resistance test involves dissolving the sample in a good solvent, adding the solvent, stirring the precipitated solid briefly, and then filtering it immediately.

[0064] 4. The cooling crystallization experiment involves dissolving a certain amount of sample into a corresponding solvent at high temperature, and then directly stirring and crystallizing at room temperature or low temperature.

[0065] 5. The polymer template experiment involves adding different types of polymer materials to a clear solution of the sample and leaving it at room temperature to evaporate until the solvent is dry.

[0066] 6. Thermal method experiments involve treating the sample under specific thermal crystallization conditions and then cooling it to room temperature.

[0067] 7. The water vapor diffusion experiment involves placing the sample in an environment with a certain humidity at room temperature.

[0068] The terms "good solvent" and "bad solvent" in this invention are relative. In a pair of solvents, the one with higher solubility is a good solvent, and the one with lower solubility is a bad solvent.

[0069] Unless otherwise specified, the solvent used in the above preparation method may be a single solvent or a combination of two or more solvents.

[0070] The X-ray powder diffraction or DSC pattern and TGA pattern disclosed in this invention, which are substantially the same, also fall within the scope of this invention.

[0071] Unless otherwise stated, the terms used in the specification and claims have the following meanings.

[0072] As used in this invention, "crystal of the present invention", "crystal form of the present invention", "crystal form of the present invention" and the like are interchangeable.

[0073] The "room temperature" mentioned in this invention generally refers to 4-30℃, and preferably to 20±5℃.

[0074] The crystal structure of the present invention can be analyzed using various analytical techniques known to those skilled in the art, including but not limited to X-ray powder diffraction (XRD), differential scanning calorimetry (DSC), and / or thermogravimetric analysis (TGA), also known as thermogravimetry (TG).

[0075] The "2θ or 2θ angle" mentioned in this invention refers to the peak position expressed in degrees (°) based on the setup in an X-ray diffraction experiment, and is typically the horizontal axis unit in a diffraction pattern. If the reflected beam is diffracted when the incident beam forms an angle θ with a certain lattice plane, the experimental setup needs to record the reflected beam at a 2θ angle. It should be understood that the specific 2θ value for a particular crystal form mentioned herein is intended to represent the 2θ value (expressed in degrees) measured using the X-ray diffraction experimental conditions described herein, and the error range of the 2θ may be ±0.3, ±0.2, or ±0.1.

[0076] It is understood that the numerical values ​​described and protected in this invention are approximate. Variations within these values ​​may be attributed to equipment calibration, equipment errors, crystal purity, crystal size, sample size, and other factors.

[0077] It is understood that the crystal forms of the present invention are not limited to those that are exactly the same as the characteristic spectra described in the accompanying drawings, such as XRD, DSC, TGA, and DVS. Any crystal form having a characteristic spectra that are substantially the same or essentially the same as those described in the accompanying drawings falls within the scope of the present invention. Attached Figure Description

[0078] Figure 1 shows the X-ray powder diffraction pattern of crystal form 1 of compound (I).

[0079] Figure 2 shows the differential scanning calorimetry curve of crystal form 1 of compound (I).

[0080] Figure 3 shows the thermogravimetric analysis spectrum of crystal form 1 of compound (I).

[0081] Figure 4 shows the isothermal adsorption curve of crystal form 1 of compound (I).

[0082] Figure 5 shows the single crystal structure of crystal form 1 of compound (I).

[0083] Figure 6 shows the amorphous X-ray powder diffraction pattern of compound (I).

[0084] Figure 7 shows the differential scanning calorimetry curve of the amorphous compound of formula (I).

[0085] Figure 8 shows the thermogravimetric analysis spectrum of the amorphous compound (I).

[0086] Figure 9 shows the isothermal adsorption curves of the amorphous compound (I). Detailed Implementation

[0087] The structure of the compound was determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). NMR shifts (δ) were expressed in 10⁻¹⁰ increments. -6 The unit (ppm) is given. NMR determination was performed using a WNMR-I 400 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD). The internal standard was tetramethylsilane (TMS).

[0088] XRD measurements were performed using a Malvern Panalytical X-ray powder diffractometer. The 2θ scanning angle ranged from 4 to 40°2θ, with a scan step of 0.01°. The phototube voltage and current for the test samples were 40 kV and 40 mA, respectively, and the sample disk was a zero-background sample disk.

[0089] TGA test conditions: The thermogravimetric analyzer was a NETZSCH (DSC 214). 1-10 mg of sample was placed in a pre-equilibrated sample pan and automatically weighed inside the TGA furnace. The sample was heated to the final temperature at a rate of 10 °C / min, with nitrogen purging at a rate of 30 mL / min.

[0090] DSC test conditions: The differential scanning calorimeter was a NETZSCH (NETZSCH TG 209F3). Samples of 0.5 mg to 5 mg were accurately weighed and placed in a standard pan or a perforated aluminum crucible sample pan. The sample was heated to the final temperature at a rate of 10 °C / min, with nitrogen purging at a rate of 40 mL / min.

[0091] The known starting materials of this invention can be synthesized using methods known in the art.

[0092] Unless otherwise specified in the examples, "solution" refers to an aqueous solution.

[0093] The following detailed embodiments illustrate the implementation process and beneficial effects of the present invention, aiming to help readers better understand the essence and characteristics of the present invention, and are not intended to limit the scope of implementation of this case.

[0094] Example 1: Preparation of compound (I)

[0095] Compound (I) was prepared according to the method described in WO2023109802.

[0096] Example 2: Preparation of crystal form 1 of compound (I)

[0097] Take about 250 mg of compound (I), add 7.5 mL of methanol, stir at 25 °C, stir for 2 hours, filter, wash the filter cake with 30% ethanol aqueous solution, and dry under vacuum to obtain crystal form 1 of compound (I).

[0098] 1 H NMR (400MHz, DMSO) δ6.96(s,1H),6.85(s,2H),6.66(s,1H),6.09(t,J=5.8Hz,1H),5.38(d,J=43.7Hz,2H),4.19(t,J=6.8 Hz,2H),3.93(dd,J=18.2,12.3Hz,2H),3.80(s,3H),3.44–3.10(m,2H),2.90(t,J=6.0Hz,2H),2.22(s,3H),1.97(s,6H).

[0099] Example 3: Preparation of the amorphous form of compound (I)

[0100] Take about 250 mg of compound (I), add 7.5 mL of methanol and 22.5 mL of dichloromethane, stir at 25 °C to dissolve, filter, spray dry to amorphous form, and vacuum dry to obtain amorphous compound (I).

[0101] Crystal form test example

[0102] Table 2. Instrument Information and Testing Method Parameters

[0103] Stability study of compound crystal form 1 of formula (I) of this invention:

[0104] Under high temperature (60℃), high humidity (75%RH), high humidity (92.5%RH), and light intensity (5000lx±500lx, 90±5μw / cm²), the following conditions were observed: 2 Stability-related data (including crystal form, moisture content, purity, particle size distribution, etc.) of crystal form 1 were tested under accelerated test (40℃±2℃ / 75%RH±5%RH), long-term test (2~8℃), and long-term test (25℃±2℃ / 60%±5%RH) conditions. The results are shown in Tables 3-9.

[0105] Table 3. Results of Influencing Factors—High Temperature (60℃) Test

[0106] Table 4. Results of Influencing Factors—High Humidity (75% RH) Test

[0107] Table 5. Results of Influencing Factors—High Humidity (92.5% RH) Test

[0108] Table 6. Experimental Results of Influencing Factors - Illumination (5000lx±500lx, 90±5μw / cm²) 2 )test

[0109] Table 7. Results of accelerated (40℃±2℃ / 75%RH±5%RH) test

[0110] Table 8. Results of long-term (2-8℃) test

[0111] Table 9. Results of long-term (25℃±2℃ / 60%±5%RH) test

[0112] The results in Tables 3-9 show that the solid-state stability of crystal form 1 of compound (I) of the present invention is good under all the stability test conditions described above. Amorphous grinding experiment of compound (I) of the present invention:

[0113] Compound (I) was wet-milled in different solvents for 5 minutes and XRPD was performed. The results are shown in Table 10.

[0114] Table 10. Results of wet milling crystal transformation test

[0115] As can be seen from Table 10, under different solvent conditions, wet milling transformed the amorphous form into the more stable crystal form 1.

[0116] Specific peak characterization results of XRD tests for crystal form 1 of compound (I)

[0117] The X-ray powder diffraction (XRD) pattern of crystal form 1 of compound (I) is shown in Figure 1. The specific peak values ​​are shown in Table 11.

[0118] Table 11

[0119] Rat PK data of compound crystal form 1 of formula (I) of this invention

[0120] Four groups of rats were used: an intravenous injection group (1 mg / kg), a single inhalation group (1.036 mg / kg and 12.928 mg / kg), and a single inhalation and a medium-dose inhalation group (3.346 mg / kg, administered once daily for 7 days). Each group consisted of six rats (half male and half female). The inhalation groups used a suspension of crystal form 1. Pharmacokinetic parameters were calculated based on the drug concentration-time curves, and the results are shown in the table below (mean values ​​for both male and female rats).

[0121] Table 12 Pharmacokinetic parameters of rats after a single intravenous injection of 1 mg / kg crystal form 1 (Mean ± SD, n = 6, half male and half female)

[0122] Table 13 Pharmacokinetic parameters of rats after inhalation of different doses of crystal form 1 (Mean ± SD, n = 6, half male and half female)

[0123] Beagle PK data for crystal form 1 of compound of formula (I) of this invention

[0124] Beagles were divided into four groups: an intravenous injection group (0.2 mg / kg), a single inhalation group (0.175 mg / kg and 1.648 mg / kg), and a single inhalation and subsequent 7-day (once daily) medium-dose inhalation group (0.600 mg / kg). Each group consisted of six rats, half male and half female. The inhalation groups used a suspension of crystal form 1. Pharmacokinetic parameters were calculated based on the drug concentration-time curves, and the results are shown in the table below (mean values ​​for male and female rats).

[0125] Table 14 Pharmacokinetic parameters of beagle dogs after a single intravenous injection of 0.2 mg / kg crystal form 1 (Mean ± SD, n = 6, half male and half female)

[0126] Table 15 Pharmacokinetic parameters of different doses of crystal form 1 administered by inhalation to beagle dogs (Mean ± SD, n = 6, half male and half female)

[0127] In vitro liver microsomal metabolic stability

[0128] This study used liver microsomes from five species—mouse, rat, dog, monkey, and human—as in vitro models to evaluate the metabolic stability of crystal form 1 of compound (I). At 37°C, 1 μM of crystal form 1 of compound (I) was incubated with liver microsomes from the five species using an NADPH regeneration system for different times (5, 15, 30, 45, 60 min). The concentration of compound (I) in the resulting samples was determined using LC-MS / MS.

[0129] The residual rates of compound (I) crystal form 1 in human, monkey, dog, rat, and mouse liver microsomes after incubation were 10.6% (60 min), 0.328% (30 min), 0.296% (45 min), 0.261% (45 min), and 0.437% (30 min), respectively. In the negative control, the concentration of compound (I) crystal form 1 in inactivated human, monkey, dog, rat, and mouse liver microsomes did not significantly decrease after 60 min of incubation. The intrinsic clearance rate (CL) of human, monkey, dog, rat, and mouse liver was calculated. int The values ​​were 98.6, 540, 598, 453 and 1324 mL / min / kg, respectively, and the liver clearance rate (CL) was... hb The values ​​were 17.1, 40.7, 29.4, 49.2 and 84.3 mL / min / kg, respectively.

[0130] In summary, based on the results of the liver microsomal metabolic stability test, it is inferred that compound (I) belongs to the class of highly cleared drugs in humans, monkeys, dogs, rats, and mice.

Claims

1. A crystalline form of a compound of formula (I), 2. The crystalline form as described in claim 1, wherein it is crystalline form 1 of the compound of formula (I), characterized in that, Crystal form 1 is a triclinic crystal system, and the crystal has no helical axis. Space group, cell constant is α = 91.511(2)°, γ = 90.364(2)°, β = 99.668(2)°, cell volume The Z' of the system is 2.

3. The crystalline form of the compound as described in claim 1, wherein the crystalline form 1 of the compound of formula (I) is subjected to Cu-Kα radiation, and its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ positions: 10.35°±0.2°, 13.12°±0.2°, 15.56°±0.2°, 17.95°±0.2°, and 21.34°±0.2°.

4. The crystalline form as described in claim 3, when subjected to Cu-Kα radiation, exhibits characteristic diffraction peaks at the following 2θ positions in its X-ray powder diffraction pattern: 10.35°±0.2°, 12.83°±0.2°, 13.12°±0.2°, 14.99°±0.2°, 15.56°±0.2°, 17.95°±0.2°, 21.34°±0.2°, and 22.81°±0.2°.

5. The crystalline form as described in claim 4, when subjected to Cu-Kα radiation, exhibits characteristic diffraction peaks at the following 2θ positions in its X-ray powder diffraction pattern: 6.54°±0.2°, 10.35°±0.2°, 12.83°±0.2°, 13.12°±0.2°, 14.47°±0.2°, 14.99°±0.2°, 15.56°±0.2°, 16.12°±0.2°, 17.95°±0.2°, 21.34°±0.2°, 22.81°±0.2°, 23.25°±0.2°, 29.18°±0.2°, and 31.53°±0.2°.

6. The crystalline form as described in claim 5, when subjected to Cu-Kα radiation, has an X-ray powder diffraction pattern that is essentially as shown in Figure 1.

7. A pharmaceutical composition comprising a therapeutically effective amount of the crystal form according to any one of claims 1-6, and a pharmaceutically acceptable carrier and / or excipient.

8. Use of the crystalline form of any one of claims 1-6, or the pharmaceutical composition of claim 7, in the preparation of a medicament for treating / preventing PDE3 / 4 mediated diseases.

9. A method for treating / preventing PDE3 / 4-mediated diseases, the method comprising administering to a subject a therapeutically effective amount of the crystalline form of any one of claims 1-6, or the pharmaceutical composition of claim 7.

Images