Crystalline form of substituted 2-(pyrrolidin-3-yl) acetate derivative, preparation method therefor, and use thereof

Abstract

The present invention provides a solid form, a crystalline form, a crystal form A, a crystal form B, a crystal form C, and a salt form of a compound represented by formula (I), a preparation method therefor, and a use of the compound represented by formula (I) in the preparation of a drug for preventing and / or treating a disease or condition associated with elevated plasma Lp(a) levels.

Description

Crystal form of a substituted 2-(pyrrolidine-3-yl)acetic acid derivative, its preparation method and uses Technical Field

[0001] This invention relates to the field of pharmaceutical technology, and more particularly to compounds that are Lp(a) inhibitors, especially crystalline forms, salt forms, preparation methods, and uses of substituted 2-(pyrrolidine-3-yl)acetic acid derivatives. Background Technology

[0002] Lipoprotein(a) (Lp(a)) is a lipoprotein particle synthesized in the liver, consisting of an apolipoprotein(a) (Apo(a)) and a low-density lipoprotein-like particle containing an apolipoprotein(b) (Apo(b)).

[0003] The level of Lp(a) is mainly determined by genes, varies significantly among different populations, and is almost unaffected by lifestyle interventions.

[0004] Lp(a) is associated with an increased risk of coronary artery disease, ischemic stroke, aortic stenosis, heart failure, atrial fibrillation, and peripheral artery disease. Approximately 20% of the population has elevated serum Lp(a) levels (≥30 mg / dL). The increased risk of cardiovascular disease (CVD) associated with Lp(a) is primarily attributed to the dual procoagulant effect of Apo(a), and the atherogenic (AS) and pro-inflammatory effects of Apo B containing oxidized phospholipid components (OxPL). Lp(a) is a contributing factor not only to atherosclerotic cardiovascular disease (ASCVD) but also to calcific aortic valve disease. Elevated plasma Lp(a) levels are an independent risk factor for CVD.

[0005] For patients with elevated Lp(a) levels, approved treatment options are very limited. Apheresis can be used to filter blood to remove LDL and Lp(a); however, the effects are temporary and usually need to be repeated every two weeks, and patient adherence is not good. To date, there are no approved drug therapies for lowering Lp(a) levels. Therefore, it is essential to provide patients with CVD with pharmaceutically acceptable compounds and treatment options to lower plasma Lp(a) levels. Summary of the Invention

[0006] In a first aspect, the present invention provides the solid form of the compound of formula (I):

[0007] Secondly, the present invention provides a crystalline form of the compound of formula (I).

[0008] Thirdly, the present invention provides a crystal form C of the compound of formula (I), characterized in that, using Cu-Kα radiation, X-ray powder diffraction in terms of 2θ angle has characteristic peaks at 4.4±0.2° and 17.6±0.2°.

[0009] In some embodiments, the crystal form C of the compound of formula (I), when subjected to Cu-Kα radiation, exhibits characteristic peaks at 4.4±0.2°, 8.8±0.2°, and 17.6±0.2° in X-ray powder diffraction at an angle of 2θ.

[0010] In some embodiments, the crystal form C of the compound of formula (I), when subjected to Cu-Kα radiation, exhibits characteristic peaks at 4.4±0.2°, 8.8±0.2°, 13.2±0.2°, 17.6±0.2°, and 18.4±0.2° in X-ray powder diffraction at 2θ angles.

[0011] In some embodiments, the crystal form C of the compound of formula (I), when subjected to Cu-Kα radiation, exhibits characteristic peaks at 4.4±0.2°, 5.6±0.2°, 8.8±0.2°, 13.2±0.2°, 17.6±0.2°, and 18.4±0.2° in X-ray powder diffraction at 2θ angles.

[0012] In some embodiments, the crystal form C of the compound of formula (I), when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at angles of 4.4±0.2°, 5.6±0.2°, 8.8±0.2°, 13.2±0.2°, 16.2±0.2°, 17.6±0.2°, and 18.4±0.2°.

[0013] In some embodiments, the crystal form C of the compound of formula (I), when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at angles of 4.4±0.2°, 5.6±0.2°, 8.0±0.2°, 8.8±0.2°, 13.2±0.2°, 16.2±0.2°, 17.6±0.2°, and 18.4±0.2°.

[0014] In some embodiments, the crystal form C of the compound of formula (I), when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at 4.4±0.2°, 5.6±0.2°, 6.1±0.2°, 8.0±0.2°, 8.8±0.2°, 13.2±0.2°, 16.2±0.2°, 16.7±0.2°, 17.6±0.2°, 18.4±0.2°, 19.2±0.2°, and 21.2±0.2°.

[0015] In some embodiments, the crystal form C of the compound of formula (I) has an X-ray powder diffraction (XRPD) pattern substantially as shown in Figure 1 or Figure 4.

[0016] In some embodiments, the differential scanning calorimetry (DSC) spectrum of the compound of formula (I) in crystal form C shows a first endothermic peak below 100 °C and a second endothermic peak at 270 °C ± 5 °C.

[0017] In some embodiments, the differential scanning calorimetry (DSC) spectrum of the crystal form C of the compound of formula (I) shows a first endothermic peak at 60–100 °C and a second endothermic peak at 270 °C ± 5 °C. The first endothermic peak is preferably at 60–90 °C, more preferably at 70–90 °C.

[0018] Fourthly, the present invention also provides a crystal form A of the compound of formula (I), characterized in that, using Cu-Kα radiation, X-ray powder diffraction in terms of 2θ angle has characteristic peaks at 17.4±0.2°, 19.2±0.2°, and 21.1±0.2°.

[0019] In some embodiments, the crystal form A of the compound of formula (I), when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at angles of 2θ at 8.5±0.2°, 12.9±0.2°, 13.8±0.2°, 16.3±0.2°, 17.4±0.2°, 19.2±0.2°, and 21.1±0.2°.

[0020] In some embodiments, the crystal form A of the compound of formula (I), when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at angles of 2θ at 5.3±0.2°, 8.5±0.2°, 12.9±0.2°, 13.8±0.2°, 16.3±0.2°, 17.4±0.2°, 19.2±0.2°, and 21.1±0.2°.

[0021] In some embodiments, the crystal form A of the compound of formula (I) has an X-ray powder diffraction (XRPD) pattern substantially as shown in Figure 2.

[0022] In some embodiments, the differential scanning calorimetry (DSC) spectrum of the compound of formula (I) in crystal form A shows endothermic signal peaks at approximately 73±10 °C, 201±5 °C, and 262±5 °C.

[0023] Fifthly, the present invention also provides crystal form B of the compound of formula (I), characterized in that, using Cu-Kα radiation, X-ray powder diffraction in terms of 2θ angle has characteristic peaks at 7.6±0.2°, 11.5±0.2°, 17.8±0.2°, and 19.1±0.2°.

[0024] In some embodiments, the crystal form B of the compound of formula (I), when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at 7.6±0.2°, 11.5±0.2°, 15.3±0.2°, 17.8±0.2°, 19.1±0.2°, 20.8±0.2°, 22.3±0.2°, and 23.0±0.2° in 2θ angles.

[0025] In some embodiments, the crystal form B of the compound of formula (I), when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at 7.6±0.2°, 11.5±0.2°, 15.3±0.2°, 16.1±0.2°, 16.7±0.2°, 17.8±0.2°, 19.1±0.2°, 20.8±0.2°, 22.3±0.2°, and 23.0±0.2° in 2θ angles.

[0026] In some embodiments, the crystal form B of the compound of formula (I) has an X-ray powder diffraction (XRPD) pattern substantially as shown in Figure 3.

[0027] In some embodiments, the crystal form B of the compound of formula (I) shows endothermic signals at approximately 47±5°C, 84±5°C, 161±5°C and 272±5°C according to DSC results.

[0028] In a sixth aspect, the present invention also provides a method for preparing crystal form C of the compound of formula (I), comprising the following steps:

[0029] Step 1: Add the crude product of compound (I) to the first solvent;

[0030] Step 2: Add the second solvent, heat to 40 to 60°C, and maintain the temperature for crystallization;

[0031] Step 3: Filter, collect the filter cake, and dry it to obtain the crystal form C of compound (I).

[0032] In some embodiments of the present invention, the first solvent may be selected from water or trifluoroethanol, preferably from water.

[0033] In some embodiments of the present invention, the second solvent is a poor solvent for the compound of formula (I), preferably isopropanol, tert-butanol, DMF (dimethylformamide), and more preferably DMF.

[0034] In some embodiments of the present invention, the volume ratio of the first solvent to the second solvent is 1:0.5 to 1:2, preferably 1:1 to 1.5, and more preferably 1:1.2 to 1.3.

[0035] In some embodiments of the present invention, step three, before filtration, may include the optional addition of a third solvent and a cooling and crystallization process, wherein the third solvent is a poor solvent for the compound of formula (I), the third solvent is preferably acetone, and the cooling and crystallization temperature is preferably 20-30°C.

[0036] In some embodiments of the present invention, the heating temperature in step two is 45-50°C.

[0037] In some embodiments of the present invention, the heat preservation and crystallization time in step two is 10 to 15 hours.

[0038] In a seventh aspect, the present invention also provides salts of compounds of formula (I), said salts being selected from: succinates, L-tartrates, citrates, fumarates, zinc salts, calcium salts, magnesium salts, benzenesulfonates, benzoates, adipates, L-malates, maleates, D-tartrates, oxalates, phosphates, hydrochlorides, and sulfates.

[0039] In some embodiments of the present invention, the salt is selected from: L-tartrate, citrate, succinate, fumarate, and zinc salt.

[0040] In an eighth aspect, the present invention also provides a pharmaceutical composition comprising a solid form, crystalline form, crystal form C, crystal form A, crystal form B, or a mixture of any two or more crystal forms C, crystal form A, and crystal form B, or a salt of a compound of formula (I) as described in the seventh aspect, and optionally a pharmaceutically acceptable carrier.

[0041] The above-described pharmaceutical composition is formulated into clinically acceptable preparations, such as oral preparations, injectable preparations, topical preparations, and excipient preparations, with oral preparations being preferred. The oral preparations are preferably solid dosage forms, such as tablets, capsules, and granules. These preparations can be obtained using appropriate excipients known to those skilled in the art and employing correspondingly known pharmaceutical preparation techniques.

[0042] In a ninth aspect, the present invention also provides the use of the solid form, crystalline form, crystal form A, crystal form B, crystal form C of the compound of formula (I) described in the first to fifth aspects, or a salt of the compound of formula (I) described in the seventh aspect, or the pharmaceutical composition described in the eighth aspect, in the preparation of a medicament for the prevention and / or treatment of cardiovascular disease (CVD).

[0043] In one embodiment of the invention, use is provided in the preparation of a medicament for the prevention and / or treatment of a disease or condition associated with elevated Lp(a) plasma levels, in solid form, crystalline form, crystal form A, crystal form B, crystal form C of a compound of formula (I) as described in the first to fifth aspects, or a salt of a compound of formula (I) as described in the seventh aspect, or a pharmaceutical composition as described in the eighth aspect.

[0044] In one embodiment of the present invention, the disease or condition associated with elevated Lp(a) plasma levels is cardiovascular disease (CVD); the cardiovascular disease (CVD) includes, but is not limited to, atherosclerotic cardiovascular disease (ASCVD), coronary artery stenosis, coronary artery-related disease, aortic stenosis, aortic stenosis-related disease, heart failure, heart failure-related disease, atrial fibrillation, and atrial fibrillation-related disease; the ASCVD includes peripheral vascular disease, peripheral artery-related disease, coronary heart disease, ischemic stroke, and ischemic stroke-related disease.

[0045] The term “elevated Lp(a) plasma level” refers to a plasma level that is equal to or higher than normal. For humans, “elevated Lp(a) plasma level” means a plasma level that is equal to or higher than approximately 30 mg / dL.

[0046] In one embodiment of the invention, for humans, "elevated Lp(a) plasma levels" is defined as plasma levels equal to or higher than approximately 50 mg / dL. The compounds provided herein can be used for treatment to lower Lp(a) plasma levels.

[0047] Definitions and Explanations

[0048] Unless otherwise stated, the following terms and phrases used herein are intended to have the following meanings. A particular phrase or term should not be considered uncertain or unclear unless specifically defined, but should be understood in its ordinary sense.

[0049] The solid form of the compound of formula (I) mentioned in this application includes both the amorphous and crystalline forms of the compound of formula (I).

[0050] The crystalline form of the compound of formula (I) mentioned in this application includes the anhydrous and solvent-free form, hydrate form, solvate form and eutectic form of the compound of formula (I).

[0051] The term "room temperature" refers to room temperature in the conventional sense of the art, generally 10 to 30°C, preferably 25°C ± 5°C.

[0052] In X-ray powder diffraction (XRD) patterns, the term "substantially" or "substantially as shown" refers to a substantially pure crystal form in which at least 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99% of the peaks appear in the given pattern. Furthermore, as the content of a crystal form in a product gradually decreases, some diffraction peaks attributable to that crystal form in the XRD pattern may decrease due to factors such as the instrument's detection sensitivity. Additionally, for any given crystal form, the peak positions may have slight errors, which is well known in crystallography. For example, peak positions can shift due to variations in temperature during sample analysis, sample movement, or instrument calibration; the measurement error for the 2θ value is sometimes approximately ±0.3°, and typically approximately ±0.2°. Therefore, this error should be taken into account when determining each crystal structure, and the terms "substantially" or "substantially as shown in the figure" are also intended to cover such differences in diffraction peak positions, referring to ±0.3°, preferably ±0.2°.

[0053] As used herein, the term "prevention" means, when used for a disease or condition (e.g., cardiovascular disease), that the compound or drug reduces the frequency of symptoms of the medical condition or delays the onset of the condition in a subject compared to a subject who has not been given the compound or drug (e.g., the combination product claimed in this application).

[0054] As used in this article, the term “treatment” means to reduce, alleviate or improve the symptoms of a disease or condition, improve underlying metabolic symptoms, suppress a disease or condition, such as preventing the development of a disease or condition, alleviating a disease or condition, causing the remission of a disease or condition, relieving the condition caused by a disease or condition, or preventing the symptoms of a disease or condition.

[0055] The terms "pharmaceuticalally acceptable carrier" or "pharmaceuticalally acceptable excipient" refer to carriers or excipients that do not cause significant irritation to the organism and do not impair the biological activity and properties of the active compound.

[0056] All solvents used in this application are commercially available and can be used without further purification. Beneficial effects

[0057] The crystal form of the compound of formula (I) provided by the present invention, especially crystal form C, has high crystallinity, good solubility, stable quality, and is easy to prepare into a drug.

[0058] Compound (I) exhibits strong inhibitory effects on Lp(a) assembly, good pharmacokinetic characteristics, and can significantly reduce serum Lp(a) levels in mice. Attached Figure Description

[0059] Figure 1 shows the XRPD spectrum of crystal form C of the compound of formula (I) obtained in Example 1.

[0060] Figure 2 shows the XRPD spectrum of crystal form A of the compound of formula (I) obtained in Example 3.

[0061] Figure 3 shows the XRPD spectrum of crystal form B of the compound of formula (I) obtained in Example 4.

[0062] Figure 4 shows the XRPD spectrum of crystal form C of the compound of formula (I) obtained in Example 2.

[0063] Figure 5 shows the DSC curve of crystal form C of the compound of formula (I) obtained in Example 2. Detailed Implementation

[0064] The technical solution of the present invention will be further described in detail below with reference to specific embodiments. The following embodiments are merely illustrative and explanatory of the present invention and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection intended by the present invention.

[0065] Unless otherwise stated, the raw materials and reagents used in the following examples are commercially available products or can be prepared by known methods.

[0066] In the following examples, the analytical methods used are as follows: nuclear magnetic resonance (¹H NMR), X-ray powder diffraction (XRPD), thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), and dynamic moisture adsorption-desorption analysis (DVS):

[0067] 1. Nuclear magnetic resonance analysis (¹H NMR)

[0068] Several milligrams of solid sample were dissolved in deuterated aqueous solvent and analyzed by nuclear magnetic resonance on a Bruker AVANCE NEO 400 (Bruker, GER).

[0069] 2. X-ray powder diffraction (XRPD)

[0070] The methods used in Examples 1 / 3 / 4 are as follows: The solid samples obtained in the experiments were analyzed using a Panalytical EMPYREAN (PANalytical, UK) X-ray powder diffractometer. The 2θ scanning angle ranged from 3° to 45°, the scanning step size was 0.013°, and the total testing time was 3.5 min. The testing method was Cu target Kα1 X-ray, voltage 45 kV, current 40 mA, and a zero-background sample disk.

[0071] Example 2 employed the following method: The obtained solid sample was analyzed using a SmartLabSE (Rigaku, JP) X-ray diffractometer. The 2θ scanning angle ranged from 3° to 40°, with a scanning step size of 0.02°, and a total testing time of 3.7 min. The testing method was Cu target Kβ radiation, with a voltage of 40 kV, a current of 40 mA, and a zero-background sample disk.

[0072] 3. Differential Scanning Calorimetry (DSC)

[0073] The methods used in Examples 1 / 3 / 4 are as follows: The differential scanning calorimeter was a TA Discovery 250 (TA, US). 1-2 mg of sample was accurately weighed and placed in a perforated DSC Tzero sample pan. The sample was heated to the final temperature at a rate of 10 °C / min, with nitrogen purging at a rate of 50 mL / min.

[0074] Example 2 uses the following method: The differential scanning calorimeter is a Netzsch DSC 3500 (Netzsch, Germany). Approximately 2 mg of sample was accurately weighed and placed in the DSC crucible. The crucible lid was then tightened, and a small hole was punched in the lid. The crucible was placed in the furnace, along with a reference crucible. The sample was heated to 320°C at a rate of 10°C / min, with nitrogen purging at a rate of 100 mL / min.

[0075] 4. Dynamic moisture adsorption-desorption analysis (DVS)

[0076] Preliminary assessment of hygroscopicity (simple dynamic moisture adsorption-desorption analysis) was performed using DVS Intrinsic / DVS Intrinsic Plus (SMS, UK). The test was conducted in gradient mode. After the test, XRPD analysis was performed on the sample to confirm whether the solid form had changed.

[0077] Preparation Example 1: Preparation of Compound (I)

[0078] Compounds of formula (I) can be prepared by the following method:

[0079] Step 1: Preparation of tert-butyl (R)-3-((S)-1-(tert-butoxy)-1-oxo-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborhecyclopentan-2-yl)phenyl)prop-2-yl)pyrrolidine-1-carboxylic acid:

[0080] In a single-necked flask, (R)-3-((S)-3-(3-bromophenyl)-1-(tert-butoxy)-1-oxopropane-2-yl)pyrrolidine-1-carboxylic acid tert-butyl ester (1.5 g, 3.30 mmol, 1.0 eq) was dissolved in 1,4-dioxane (20 mL), followed by the addition of 1,1-bis(diphenylphosphine)ferrocene palladium dichloromethane dichloride complex (II) (270.0 mg, 0.33 mmol, 0.1 eq), pinacol diborate (1.26 g, 4.95 mmol, 1.5 eq), and potassium acetate (972 mg, 9.9 mmol, 3 eq). The mixture was stirred at 90 °C for 16 hours under nitrogen protection. Liquid chromatography-mass spectrometry (LC-MS) analysis confirmed the reaction was complete. The reaction solution was directly concentrated and purified by rapid chromatography (Silica gel, PE:EtOAc = 2:1) to obtain the target compound (1.2 g). LCMS(ESI)[M-Boc+H]+=402.2.

[0081] Step 2: Preparation of tert-butyl (R)-3-((S)-1-(tert-butoxy)-3-(3-hydroxyphenyl)-1-oxopropane-2-yl)pyrrolidine-1-carboxylic acid:

[0082] Take a single-necked flask and dissolve the product obtained in the first step (1.2 g, 2.39 mmol, 1.0 eq) in tetrahydrofuran (20 mL), then add hydrogen peroxide (0.5 mL) and stir at room temperature for 16 hours. Liquid chromatography-mass spectrometry (LC-MS) showed the reaction was complete. The reaction solution was directly concentrated and purified by rapid chromatography (Silica gel, PE:EtOAc = 1:1) to obtain the target compound (800.0 mg). LC-MS (ESI) [M-2x tert-butyl+H]+ = 280.0.

[0083] Step 3: Preparation of tert-butyl (R)-3-((S)-1-(tert-butoxy)-3-(3-(2-(1,3-dioxoisoindoline-2-yl)ethoxy)phenyl)-1-oxopropane-2-yl)pyrrolidine-1-carboxylic acid:

[0084] Take a single-necked flask and dissolve the product obtained in step two (500.0 mg, 1.28 mmol, 1.0 eq) in N,N-dimethylformamide (5 mL). Then add 2-(2-bromoethyl)isoindoline-1,3-dione (647.7 mg, 2.56 mmol, 2 eq) and potassium carbonate (530.7 mg, 3.83 mmol, 3 eq). Stir at 90 °C for 2 hours. Liquid chromatography-mass spectrometry (LC-MS) indicates the reaction is complete. The reaction solution is directly concentrated and purified by rapid chromatography (Silica gel, PE:EtOAc = 1:1) to obtain the target compound (280 mg). LC-MS (ESI) [M+H]+ = 565.3.

[0085] Step 4: Preparation of tert-butyl (R)-3-((S)-3-(3-(2-aminoethoxy)phenyl)-1-(tert-butoxy)-1-oxopropane-2-yl)pyrrolidine-1-carboxylic acid:

[0086] Take a single-necked flask and dissolve the product obtained in step three (280.0 mg, 0.5 mmol, 1.0 eq) in ethanol (10 mL), then add hydrazine hydrate (124 mg, 2.50 mmol, 5.0 eq), and react at 90 °C for 16 hours. Liquid chromatography-mass spectrometry (LC-MS) indicates the reaction is complete. The reaction solution is directly concentrated and purified by rapid chromatography (Silica gel, PE:EtOAc = 1:5) to obtain the target compound (160 mg). LC-MS (ESI) [M+H]+ = 435.3.

[0087] Step 5: Preparation of tert-butyl pyrrolidine-1-carboxylic acid (R)-3-((S)-1-(tert-butoxy)-3-(3-(((2-(3-(((S)-3-(tert-butoxy)-2-((R)-1-(tert-butoxycarbonyl)pyrrolidine-3-yl)-3-oxopropyl)phenoxy)ethyl)amino)methyl)phenyl)-1-oxopropane-2-yl)pyrrolidine-1-carboxylic acid:

[0088] Take a single-necked flask and dissolve the product obtained in step four (160 mg, 0.37 mmol, 1.0 eq) in methanol (10 mL). Then add (R)-3-((S)-1-(tert-butoxy)-3-(3-formylphenyl)-1-oxopropane-2-yl)pyrrolidine-1-carboxylic acid tert-butyl ester (178.1 mg, 0.44 mmol, 1.2 eq), followed by sodium cyanoborohydride (23.2 mg, 0.37 mmol, 1.0 eq). Stir at room temperature for 16 hours. Liquid chromatography-mass spectrometry (LC-MS) indicates the reaction is complete. The reaction solution is directly concentrated and purified by rapid chromatography (Silica gel, PE:EtOAc = 1:5) to obtain the target compound (233 mg). LC-MS (ESI) [M+H]+ = 822.6.

[0089] Step 6: Preparation of tert-butyl (R)-3-((S)-3-(3-allylphenyl)-1-(tert-butoxy)-1-oxopropane-2-yl)pyrrolidine-1-carboxylic acid:

[0090] Take a single-necked flask and dissolve (R)-3-((S)-3-(3-bromophenyl)-1-(tert-butoxy)-1-oxopropane-2-yl)pyrrolidine-1-carboxylic acid tert-butyl ester (7.0 g, 15.4 mmol, 1.0 eq) in 1,4-dioxane (20 mL) and water (4 mL). Then add 2-allyl-4,4,5,5-tetramethyl-1,3,2-dioxaborhexacyclopentane (5.2 g, 30.8 mmol, 2 eq), potassium phosphate (9.8 g, 46.2 mmol, 3 eq), and 1,1-bis(diphenylphosphine)ferrocene palladium dichloromethane complex (II) (1.25 g, 1.54 mmol, 0.1 eq). Under nitrogen protection, stir at 90 °C for 16 hours. The reaction was confirmed to be complete by liquid chromatography-mass spectrometry (LC-MS). The reaction solution was directly concentrated and purified by rapid chromatography (Silica gel, PE:EtOAc = 2:1) to obtain the target compound (5.0 g). LC-MS [M-2×tert-butyl+H]+ = 304.1.

[0091] Step 7: Preparation of 2-(3-((S)-3-(tert-butoxy)-2-((R)-1-(tert-butoxycarbonyl)pyrrolidine-3-yl)-3-oxopropyl)phenyl)acetic acid:

[0092] Take a single-necked flask and dissolve the product obtained in step 6 (6.0 g, 14.4 mmol, 1.0 eq) in 1,4-dioxane (100 mL). Then add sodium periodate (12.3 g, 57.75 mmol, 4 eq) and potassium osmium tetroxide dihydrate (532.0 mg, 1.44 mmol, 0.1 eq). Stir at room temperature for 16 hours. Liquid chromatography-mass spectrometry (LC-MS) indicates the reaction is complete. The reaction solution is directly concentrated and purified by rapid chromatography (Silica gel, PE:EtOAc = 1:1) to obtain the target compound (500 mg). LC-MS (ESI) [M+H]+ = 434.2.

[0093] Step 8: Preparation of tert-butyl pyrrolidine-1-carboxylate (R)-3-((S)-1-(tert-butoxy)-3-(3-((N-(2-(3-((S)-3-(tert-butoxy)-2-((R)-1-(tert-butoxycarbonyl)pyrrolidine-3-yl)-3-oxopropyl)phenoxy)ethyl)-2-(3-((S)-3-(tert-butoxy)-2-((R)-1-(tert-butoxycarbonyl)pyrrolidine-3-yl)-3-oxopropyl)phenyl)acetamyl)methyl)phenyl)-1-oxopropane-2-yl)pyrrolidine-1-carboxylate:

[0094] Take a single-necked flask and dissolve the product obtained in step 5 (70 mg, 0.0852 mmol, 1.0 eq) in DMF (5 mL). Then add the product obtained in step 7 (44.3 mg, 0.102 mmol, 1.2 eq), N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (64.7 mg, 0.17 mmol, 2.0 eq), and N,N-diisopropylethylamine (33.0 mg, 0.25 mmol, 3.0 eq). Stir at room temperature for 16 hours, and the reaction is complete as determined by LC-MS. The reaction solution is directly concentrated and purified by rapid chromatography (Silica gel, PE:EtOAc = 1:8) to obtain the target compound (70 mg). LCMS (ESI) [M+H]+ = 1237.9.

[0095] Step 9: Preparation of (S)-3-(3-(2-((3-((S)-2-carboxy-2-((R)-pyrrolidine-3-yl)ethyl)benzyl)(2-(3-((S)-2-carboxy-2-((R)-pyrrolidine-3-yl)ethyl)phenoxy)ethyl)amino)-2-oxoethyl)phenyl)-2-((R)-pyrrolidine-3-yl)propionic acid:

[0096] Take a single-necked flask and dissolve the product obtained in step 8 (70.0 mg, 0.056 mmol) in dioxane hydrochloride (2 mL). Stir at room temperature for 16 hours. Liquid chromatography-mass spectrometry (LC-MS) indicates the reaction is complete. The reaction solution is directly concentrated and purified by preparative chromatography to obtain the target compound (27.2 mg). LC-MS (ESI) [M+H]+ = 769.4. 1H NMR (400 MHz, Deuterium) Oxide)δ7.20(dq,J=30.0,7.5Hz,3H),7.11-6.86(m,6H),6.81(s,1H),6.69-6.59(m,2H),4.59( d,J=4.6Hz,2H),4.10(t,J=5.0Hz,1H),4.00(d,J=5.2Hz,1H),3.88(s,1H),3.79(d,J=5.2Hz,1H ),3.69(d,J=5.2Hz,2H),3.29(dtt,J=13.1,9.3,4.6Hz,6H),3.12(dd,J=11.9,7.9Hz,3H),2.87 -2.58(m,8H),2.54-2.48(m,1H),2.41-2.23(m,6H),2.01(s,3H),1.63(dt,J=22.4,11.3Hz,3H).

[0097] Example 1: Preparation of crystal form C of compound (I)

[0098] 500.3 mg of compound (I) was weighed and added to 27.5 mL of water / acetonitrile (1 / 10, v / v). The mixture was suspended and stirred at 50 °C for 1 day. The suspension was then centrifuged, and the solid was dried under vacuum at 50 °C for 3 days to obtain 479.5 mg of white solid powder (yield 95.8%). The obtained solid was characterized by XRPD, and the results showed that it was crystalline form C.

[0099] XRPD results show that crystal form C is a well-crystallized solid. The spectral analysis data are shown in the table below:

[0100] DSC results showed that crystal form C had endothermic signals at around 77.2℃ and 268.1℃.

[0101] Example 2: Preparation of crystal form C of compound (I)

[0102] Take 50.0 g of compound (I) as raw material, add purified water (700 ml) to dissolve, add DMF (900 mL) and seed crystals (0.7 g); then heat to 45-50℃ and keep warm to crystallize for 15 h; add acetone, cool to 25±5℃ and continue to crystallize for 2 h; filter, collect filter cake, dry to obtain compound (I) crystal form C (44.7 g), yield 89.4%.

[0103] The XRPD characterization results of the obtained crystal form C are shown in the table below and Figure 4. The XRPD results show that crystal form C is a solid with good crystallinity.

[0104] The DSC results (see Figure 5) show that crystal form C has endothermic signals at around 86.4℃ and 272.4℃.

[0105] The experiment was repeated without adding acetone during the reaction. The solid obtained by crystallization was analyzed by XRPD, and the results showed that it was also crystal form C.

[0106] Example 3: Preparation of crystal form A of compound (I)

[0107] Weigh 699.8 mg of compound (I), add 0.7 ml of water at room temperature and sonicate until completely dissolved; take 0.1 ml of the solution, add 0.3 ml of dimethylformamide, stir at room temperature for 1 day, the solid precipitates, centrifuge to separate it, and then vacuum dry the solid at room temperature.

[0108] The obtained solid was characterized by XRPD, and the results showed that it was crystal form A.

[0109] The XRPD results are shown in Figure 2 and the table below, indicating that crystal form A is a solid with poor crystallinity.

[0110] DSC results showed that crystal form A had endothermic signal peaks at 72.5℃, 200.6℃ and 261.9℃.

[0111] Example 4: Preparation of crystal form B of compound (I)

[0112] 39.7 mg of compound (I) was added to 3.2 mL of a water / DMF (3 / 5, v / v) mixed solvent. The mixture was subjected to a cyclic heating and cooling process at a rate of 0.5 °C / min, alternating between 10 °C (30 min) → 50 °C (30 min) → 10 °C (30 min). After 13 cycles, the suspension was centrifuged, and the resulting solid was dried under vacuum at 50 °C. XRPD characterization of the solid revealed it to be crystal form B.

[0113] XRPD results show that crystal form B is a solid with moderate crystallinity. The results are shown in the table below and Figure 3.

[0114] DSC results showed that crystal form B had endothermic signals at approximately 47.2℃, 83.9℃, 160.6℃, and 271.9℃.

[0115] Example 5: Preparation of salts of compound (I)

[0116] The compounds of formula (I) were reacted with different ligand acids to prepare the corresponding salts:

[0117] 1. High-temperature suspension reaction

[0118] A certain amount of compound (I) and a certain equivalent of ligand were weighed and added to a certain amount of solvent. The mixture was stirred at 50°C for 3-4 hours, then transferred to room temperature and stirred for 1 day. The suspension was centrifuged, and the solid was dried under vacuum at room temperature. If the solution was clear / formed into an oil / gel, stirring was continued at room temperature for 1 day. The solution with precipitated solid was centrifuged, and the solid was dried under vacuum at room temperature. The experimental results are shown in the table below.

[0119] Table 1 Results of high-temperature suspension reaction experiment

[0120] 2. Room temperature suspension reaction

[0121] A certain amount of compound (I) and a certain equivalent of ligand were weighed and added to a certain amount of solvent. The mixture was stirred and reacted at room temperature for 2 days. The suspension was then centrifuged, and the solid was dried under vacuum at room temperature. If the solution was clear / formed into an oil / gel, the solution was stirred at 5°C for 2 days. The solution with precipitated solid was centrifuged, and the solid was dried under vacuum at room temperature. The experimental results are shown in the table below.

[0122] Table 2 Results of room temperature suspension reaction experiments

[0123] 3. Ion exchange method

[0124] A certain amount of compound sample of formula (I) and 3 equivalents of sodium hydroxide (prepared with water, 1 mol / L) were weighed and added to a certain amount of solvent. After stirring and dissolving at room temperature, 1.5 equivalents of calcium chloride (prepared with water, 1 mol / L) / magnesium chloride (prepared with water, 1 mol / L) / zinc chloride were added for ion exchange. After stirring at room temperature for 5-30 min, the suspension was centrifuged. The solid was further purified by slurrying with a certain amount of solvent, and the suspension was centrifuged again. The solid was then dried under vacuum at room temperature. The experimental results are shown in the table below.

[0125] Table 3. Experimental results of ion exchange method

[0126] A total of 17 salts of the compound of formula (I) were obtained, namely amorphous succinate, amorphous L-tartrate, amorphous citrate, amorphous fumarate, amorphous zinc salt, amorphous calcium salt, amorphous magnesium salt, amorphous benzenesulfonate, amorphous benzoate, amorphous adipate, amorphous L-malate, amorphous maleate, amorphous D-tartrate, amorphous oxalate, amorphous phosphate, amorphous hydrochloride, and amorphous sulfate.

[0127] Test Example 1: Solubility Study of Crystal Form C of Compound (I)

[0128] At room temperature (approximately 25°C), weigh approximately 20 mg of crystal form C of compound (I), add a certain amount of water gradually, stir, and observe whether the solid completely dissolves. Estimate the solubility of the compound in various solvents based on the volume of solvent required for complete dissolution. According to the solubility test results, crystal form C is readily soluble in water. The corresponding results are shown in the table below.

[0129] Table 4. Water solubility assessment results

[0130] Test Example 2: Biological and Water Solubility Test of Crystal Form C of Compound (I)

[0131] The dynamic solubility of crystal form C of compound (I) was determined at 37 °C in three biological media (FaSSIF, FeSSIF, and FaSSGF) and water. The corresponding results are shown in the table below. The results show that the 24-hour solubility in the three biological media and water is greater than 10 mg / mL, and no solid remains afterward.

[0132] Table 5 Dynamic solubility test in biological media and water

[0133] Test Example 3: Stability Study of Crystal Form C of Compound (I)

[0134] The stability of crystal form C of compound (I) was studied under conditions of 60℃, 40℃ / 75% RH and 25℃ / 60% RH. Samples were taken at 7 days for XRPD characterization and HPLC testing. The results are shown in the table below.

[0135] XRPD results showed that crystal form C did not undergo any crystal form transformation after 7 days of exposure to high temperature, accelerated conditions, and high humidity. Purity results showed that crystal form C remained chemically stable after 7 days of exposure to high temperature, accelerated conditions, and high humidity.

[0136] Table 6 Results of the stability study of crystal form C

[0137] Test Example 4: Solid-state humidity stability assessment of compound (I)

[0138] Approximately 10 mg of different crystal forms of compound (I) were weighed and placed in a drug stability test chamber at 25℃ / 60% RH, or in an atmosphere of 25℃ / 75% RH (saturated NaCl solution), or 25℃ / 85% RH (saturated KCl solution). After a period of time, samples were taken for XRPD characterization, and the results are shown in the table below. At 25℃, crystal form C remained crystal form C for 3 days at 60% RH and 75% RH; after 4 days at 85% RH, it partially transformed into crystal form B. Crystal form B partially transformed into crystal form C after 3 days at both 60% RH and 75% RH.

[0139] Table 7 Results of Solid Humidity Stability Study

[0140] *The white powder has good flowability; no gelation was observed.

[0141] Test Example 5: Solubility Evaluation of Salt Forms of Compound (I)

[0142] The solubility of five candidate salt forms—succinate amorphous, L-tartrate amorphous, citrate amorphous, fumarate amorphous, and zinc salt amorphous—in solvents was determined, and the results are shown in the table below.

[0143] Table 8. Solubility test results of each salt type

[0144] According to solubility test results, L-tartrate amorphous salts are sparingly soluble in methanol, toluene, DMF, chloroform, and THF; citrate amorphous salts are readily soluble in water but sparingly soluble in methanol, MTBE, ethylene glycol methyl ether, and acetonitrile; succinate amorphous salts are readily soluble in trifluoroethanol but sparingly soluble in methanol, acetone, ethyl acetate, n-heptane, and dioxane. In these sparingly soluble solvents, partial dissolution of the solid in methanol is observed, which is superior to other sparingly soluble solvents; fumarate amorphous salts are readily soluble in trifluoroethanol but sparingly soluble in isopropanol, acetone, cyclohexane, and dioxane; and zinc salt amorphous salts are sparingly soluble in acetonitrile, chloroform, water, DMF, and ethylene glycol methyl ether.

[0145] Test Example 6: Lp(a) Assembly Inhibition Screening ELISA Detection Experiment

[0146] 1. Reagents, consumables, and instruments

[0147] 2. Preparation of cell culture medium 2.1 Complete cell culture medium

[0148] 2.2 Experimental Culture Medium

[0149] 3. Experimental Procedure

[0150] 3.1 Cell Culture and Passaging

[0151] 1) Preheat the culture medium, DPBS, and trypsin in a 37℃ water bath.

[0152] 2) Digest cells with trypsin, transfer the cell suspension to a 15ml centrifuge tube, and centrifuge at 1000rpm for 5 minutes.

[0153] 3) Resuspend the cells in the culture medium and transfer the cell suspension to a new T75 cell culture flask.

[0154] 4) Incubate at 37℃ in a 5% CO2 incubator.

[0155] 3.2 Plate laying and chemical dosing

[0156] 1) Seed HepG2 cells overexpressing ApoA protein into 96-well plates, 100 μl per well, and culture overnight for adhesion.

[0157] 2) On the second day, replace the culture medium with fresh one, then add the diluted compound, and place the cell plate in a 5% CO2, 37°C constant temperature incubator for 24 hours.

[0158] 3.3 Preparation of cell culture supernatant

[0159] 1) Add 10 μl of 1.5 M EACA to each well.

[0160] 2) Centrifuge at 1000 rpm for 1 minute and transfer 100 μl of cell culture supernatant to a pre-coated ELISA plate.

[0161] 3.4 ELISA Experiment

[0162] 1) Add 100 μl of capture antibody to each well to pre-coat the ELISA plate and incubate overnight at 25°C.

[0163] 2) Add 300 μl of cleaning solution to each well and wash 5 times, one minute each time.

[0164] 3) Add 100 μl of blocking solution to each well and incubate at 25°C for 2 hours.

[0165] 4) Add 300 μl of cleaning solution to each well and wash 5 times, one minute each time.

[0166] 5) Add 100 μl of sample to each well and incubate at 25°C for 1 hour.

[0167] 6) Add 300 μl of cleaning solution to each well and wash 5 times, one minute each time.

[0168] 7) Add 100 μl of detection antibody to each well and incubate at 25°C for 1 hour.

[0169] 8) Add 300 μl of cleaning solution to each well and wash 5 times, one minute each time.

[0170] 9) Add 100 μl of substrate solution (Color Reagent A:Color Reagent B, 1:1) to each well and incubate at 25°C in the dark for 20 minutes.

[0171] 10) Add 50 μl of stop solution to each well, gently blow and aspirate several times with the pipette tip, and take the reading within five minutes.

[0172] 4. IC50 Data Analysis

[0173] H = Ave (DMSO or H2O)

[0174] L = Ave (1uM Reference)

[0175] SD(H) = STDEV (DMSO or H2O)

[0176] SD(L) = STDEV(1uM Reference)

[0177] CV%(DMSO)=100*(SD_H / Ave_H)

[0178] CV%(1uM Reference)=100*SD_L / Ave_L

[0179] Z'=1-3*(SD_H+SD_L) / (Ave_H-Ave_L)

[0180] Inhibition%=(Ave_H-Sample) / (Ave_H-Ave_L)

[0181] Nonlinear fitting regression equation for compound IC50:

[0182] Y=Bottom+(Top-Bottom) / (1+10^((LogIC50-X)*HillSlope))

[0183] X:cpd concentration

[0184] Y:inhibition%

[0185] Top and Bottom:Plateaus in same units as Y

[0186] logIC50: Same log units as X

[0187] HillSlope:Slope factor or Hill slope

[0188] 5. Test Results

[0189] The experimental results show that the compounds of the present invention have a strong inhibitory effect on Lp(a) assembly. Among them, LY3473329 is the compound of Example 1 in CN114008021A, and the preparation method is as described in Example 1 of patent CN114008021A.

[0190] The results are shown in the table below.

[0191] Table 9

[0192] Test Example 7: In vivo pharmacokinetic experiment in C57BL / 6J female mice

[0193] Using C57BL / 6J female mice as test animals, the drug concentration in plasma of C57BL / 6J female mice at different time points after intravenous and gavage administration of the compound was determined by LC / MS / MS, and the pharmacokinetic characteristics of the compound in C57BL / 6J female mice were studied.

[0194] Experimental animals: C57BL / 6J female mice

[0195] Drug preparation: Weigh a certain amount of the compound and add physiological saline to prepare colorless and clear solutions of 0.4 mg / mL (IV) and 1 mg / mL (PO).

[0196] Experimental Procedure: C57BL / 6J female mice were fasted overnight before being administered the drug via gavage, while intravenous administration was administered without fasting. The intravenous dose was 2 mg / kg, and the gavage dose was 10 mg / kg. Approximately 0.03 mL of blood was collected from the orbital venous plexus at 0.083 (IV), 0.25, 0.5, 1, 2, 4, 8, and 24 hours after administration. The blood samples were placed in centrifuge tubes containing EDTA-K2 and centrifuged at 3000 rpm / min for 10 min at 2-8°C to obtain plasma, which was then stored at -80°C.

[0197] Determination of the content of the target compound in the plasma of female C57BL / 6J mice at different blood collection time points: 10 μL of plasma from female C57BL / 6J mice collected at various time points after drug administration of LY3473329 and the compound of this application was added to 200 μL of 500 mM trichloroacetic acid aqueous solution to precipitate proteins, which contained 100 ng / mL internal standard. The mixture was vortexed for 1 min. Samples treated in test tubes were centrifuged at 14000 rpm for 7 min, and samples treated in 96-well plates were centrifuged at 4000 rpm for 10 min. 190 μL of supernatant was transferred to a 96-well plate, and 8 μL of supernatant was injected for LC-MS / MS analysis. Example 6 (WO2023078333A1): 12 μL of plasma from female C57BL / 6J mice collected at various time points after drug administration was added to 240 μL of methanol to precipitate proteins. The methanol contained 10 ng / mL internal standard. The mixture was vortexed for 1 min. Samples treated in test tubes were centrifuged at 14000 rpm for 7 min, and samples treated in 96-well plates were centrifuged at 4000 rpm for 10 min. 220 μL of the supernatant was transferred to a 96-well plate, and 10 μL of the supernatant was analyzed by LC-MS / MS.

[0198] Pharmacokinetic parameters

[0199] After LC / MS / MS analysis to detect blood drug concentration, pharmacokinetic parameters (such as Cmax, AUC, T1 / 2, etc.) were calculated using WinNonlin software and a non-compartmental model method.

[0200] The results showed that the compounds of this invention exhibited good pharmacokinetics, with better blood drug exposure after intravenous and gavage administration, a longer half-life, and excellent oral administration performance. Their pharmacokinetic properties, such as blood drug exposure and half-life, were superior to those of the control compounds LY3473329 and WO2023078333A1 (Compound of Example 6). The pharmacokinetic parameters of the compounds of formula (I) in C57BL / 6J female mice after gavage and intravenous administration are shown in the table below.

[0201] Among them, LY3473329 is the compound of Example 1 in CN114008021A, and the preparation method is the same as that of Example 1 in patent CN114008021A. The preparation method of the compound of Example 6 in WO2023078333A1 is the same as that of Example 6 in patent WO2023078333A1.

[0202] Table 10 C57BL / 6J female mice administered the drug via gavage. Pharmacokinetic parameters

[0203] Table 11 Pharmacokinetic parameters of intravenous administration in female C57BL / 6J mice

[0204] Test Example 8: Lp(a) Inhibition Test in Humanized Mice

[0205] The in vivo activity of the compound in reducing Lp(a) was evaluated in a humanized Lp(a) transgenic mouse model. Lp(a) mice were obtained by mating mice expressing humanized apoB 100 and mice expressing humanized apo(a) containing 16 Kringle repeats. Female mice aged 8–9 weeks were selected for the experiment. Housing conditions included a standard light cycle (12 hours light / 12 hours dark), temperature of 20–25°C, humidity of 40–70%, and free access to water and a normal diet.

[0206] The day before the experiment, mice were randomly divided into a solvent control group and a test compound group (n=5 / group) based on their body weight and baseline plasma Lp(a) concentration. Mice were orally administered either the solvent (physiological saline, 10 mL / kg) or the test compound (prepared with physiological saline to a concentration of 3 mg / mL, 10 mL / kg per mouse) once daily (9:00 am) for 3 consecutive days. Blood samples were collected after each 3-day course to measure the rate of change in serum Lp(a). Eight hours after the last administration, a 10 μL blood sample was collected from the tail and transferred to a 1.5 mL centrifuge tube containing 90 μL of physiological saline. The sample was thoroughly mixed and incubated on ice for 30 minutes. The sample was then centrifuged at 3000 rpm for 10 minutes at 4°C, and the supernatant was collected and placed on ice. A double-antibody sandwich ELISA assay was used to detect the Lp(a) concentration in the supernatant. Anti-Apo(a) capture antibody and HRP-conjugated anti-ApoB detection antibody were used. TMB was used for color development, and the reaction was terminated with 1N sulfuric acid. The OD value was read at 450 nm using a Tecan microplate reader. Calculate the inhibition rate for each treatment group. Inhibition rate (%) = (Pre-treatment Lp(a) concentration of solvent control group or self-administered drug - Post-treatment Lp(a) concentration) / Pre-treatment Lp(a) concentration of solvent control group or self-administered drug * 100.

[0207] The experimental results show that the compound of the present invention significantly reduces the serum Lp(a) level in mice in vivo.

[0208] The compounds of this invention significantly reduced serum Lp(a) levels in mice more effectively than the control compounds LY3473329 and / or WO2023078333A1, the compound of Example 6.

[0209] The inhibition rates of Lp(a) in vivo by compound (I) are shown in the table below.

[0210] Table 12. In vivo efficacy test of single-dose drugs in mice.

Claims

1. Formula (I) compound in solid form, 2. Crystalline form of the compound of formula (I).

3. The crystal form C of the compound of formula (I), characterized in that, Using Cu-Kα radiation, X-ray powder diffraction, expressed in 2θ angles, exhibits characteristic peaks at 4.4±0.2° and 17.6±0.2°, or Using Cu-Kα radiation, X-ray powder diffraction, expressed in 2θ angles, exhibits characteristic peaks at 4.4±0.2°, 8.8±0.2°, and 17.6±0.2°, or Using Cu-Kα radiation, X-ray powder diffraction, expressed in 2θ angles, exhibits characteristic peaks at 4.4±0.2°, 8.8±0.2°, 13.2±0.2°, 17.6±0.2°, and 18.4±0.2°, or Using Cu-Kα radiation, X-ray powder diffraction, expressed in 2θ angles, exhibits characteristic peaks at 4.4±0.2°, 5.6±0.2°, 8.8±0.2°, 13.2±0.2°, 17.6±0.2°, and 18.4±0.2°, or Using Cu-Kα radiation, X-ray powder diffraction, expressed in 2θ angles, exhibits characteristic peaks at 4.4±0.2°, 5.6±0.2°, 8.8±0.2°, 13.2±0.2°, 16.2±0.2°, 17.6±0.2°, and 18.4±0.2°, or Using Cu-Kα radiation, X-ray powder diffraction, expressed in 2θ angles, exhibits characteristic peaks at 4.4±0.2°, 5.6±0.2°, 8.0±0.2°, 8.8±0.2°, 13.2±0.2°, 16.2±0.2°, 17.6±0.2°, and 18.4±0.2°, or Using Cu-Kα radiation, X-ray powder diffraction, expressed in 2θ angles, exhibits characteristic peaks at 4.4±0.2°, 5.6±0.2°, 6.1±0.2°, 8.0±0.2°, 8.8±0.2°, 13.2±0.2°, 16.2±0.2°, 16.7±0.2°, 17.6±0.2°, 18.4±0.2°, 19.2±0.2°, and 21.2±0.2°. Using Cu-Kα radiation, X-ray powder diffraction patterns are obtained, which are essentially as shown in Figure 1 or Figure 4.

4. The crystal form C of the compound of formula (I) according to claim 3, characterized in that, The differential scanning calorimetry (DSC) spectrum showed a first endothermic peak below 100℃ and a second endothermic peak at 270±5℃.

5. A method for preparing crystal form C of the compound of formula (I), characterized in that, Includes the following steps: Step 1: Add the crude product of compound (I) to the first solvent; Step 2: Add the second solvent, heat to 40-60℃, and maintain the temperature for crystallization; Step 3: Filter, collect the filter cake, and dry it to obtain the crystal form C of compound (I).

6. The method for preparing crystal form C of the compound of formula (I) according to claim 5, characterized in that, The first solvent is selected from water or trifluoroethanol, preferably water.

7. The method for preparing crystal form C of the compound of formula (I) according to claim 5, characterized in that, The second solvent is selected from isopropanol, tert-butanol, and DMF, preferably DMF.

8. The method for preparing crystal form C of the compound of formula (I) according to claim 5, characterized in that, The volume ratio of the first solvent to the second solvent is 1:0.5 to 2, preferably 1:1 to 1.5, and more preferably 1:1.2 to 1.

3.

9. The method for preparing crystal form C of the compound of formula (I) according to claim 5, characterized in that, The temperature in step two is 45–50°C.

10. The method for preparing crystal form C of the compound of formula (I) according to claim 5, characterized in that, The heat preservation and crystallization time in step two is 10 to 15 hours.

11. The method for preparing crystal form C of the compound of formula (I) according to claim 5, characterized in that, The third step, before filtration, also includes optionally adding a third solvent and cooling to crystallize the material. The third solvent is selected from acetone, and the cooling and crystallization temperature is 20-30°C.

12. Crystal form A of the compound of formula (I), characterized in that, Using Cu-Kα radiation, X-ray powder diffraction, expressed in 2θ angles, exhibits characteristic peaks at 17.4±0.2°, 19.2±0.2°, and 21.1±0.2°, or Using Cu-Kα radiation, X-ray powder diffraction, expressed in 2θ angles, exhibits characteristic peaks at 8.5±0.2°, 12.9±0.2°, 13.8±0.2°, 16.3±0.2°, 17.4±0.2°, 19.2±0.2°, and 21.1±0.2°, or Using Cu-Kα radiation, X-ray powder diffraction, expressed in 2θ angles, exhibits characteristic peaks at 5.3±0.2°, 8.5±0.2°, 12.9±0.2°, 13.8±0.2°, 16.3±0.2°, 17.4±0.2°, 19.2±0.2°, and 21.1±0.2°, or Using Cu-Kα radiation, an X-ray powder diffraction pattern is obtained, which is essentially as shown in Figure 2.

13. Crystal form B of the compound of formula (I), characterized in that, Using Cu-Kα radiation, X-ray powder diffraction, expressed in 2θ angles, exhibits characteristic peaks at 7.6±0.2°, 11.5±0.2°, 17.8±0.2°, and 19.1±0.2°, or Using Cu-Kα radiation, X-ray powder diffraction, expressed in 2θ angles, exhibits characteristic peaks at 7.6±0.2°, 11.5±0.2°, 15.3±0.2°, 17.8±0.2°, 19.1±0.2°, 20.8±0.2°, 22.3±0.2°, and 23.0±0.2°, or Using Cu-Kα radiation, X-ray powder diffraction, expressed in 2θ angles, exhibits characteristic peaks at 7.6±0.2°, 11.5±0.2°, 15.3±0.2°, 16.1±0.2°, 16.7±0.2°, 17.8±0.2°, 19.1±0.2°, 20.8±0.2°, 22.3±0.2°, and 23.0±0.2°. Using Cu-Kα radiation, an X-ray powder diffraction pattern is obtained, which is essentially as shown in Figure 3.

14. A salt of the compound of formula (I) is selected from succinate, L-tartrate, citrate, fumarate, zinc salt, calcium salt, magnesium salt, benzenesulfonate, benzoate, adipate, L-malate, maleate, D-tartrate, oxalate, phosphate, hydrochloride and sulfate; preferably L-tartrate, citrate, succinate, fumarate and zinc salt.

15. A pharmaceutical composition comprising a solid form of the compound of formula (I) of claim 1, or a crystalline form of the compound of formula (I) of claim 2, or a crystal form C of any of the compounds of formula (I) of claims 3-4, or a crystal form A of the compound of formula (I) of claim 12, or a crystal form B of the compound of formula (I) of claim 13, or a mixture of any two or more of the aforementioned crystal forms C, A, and B, or a salt of the compound of formula (I) of claim 14, or a mixture of any two or more thereof, and optionally a pharmaceutically acceptable carrier.

16. Use of the solid form of the compound of formula (I) according to claim 1, or the crystalline form of the compound of formula (I) according to claim 2, or the crystal form C of any of the compounds of formula (I) according to claims 3-4, or the crystal form A of the compound of formula (I) according to claim 12, or the crystal form B of the compound of formula (I) according to claim 13, or the salt of the compound of formula (I) according to claim 14, or the pharmaceutical composition according to claim 15 in the preparation of a medicament for the prevention and / or treatment of cardiovascular disease (CVD).

17. Use of the solid form of the compound of formula (I) according to claim 1, or the crystalline form of the compound of formula (I) according to claim 2, or the crystal form C of the compound of formula (I) according to any one of claims 3-4, or the crystal form A of the compound of formula (I) according to claim 12, or the crystal form B of the compound of formula (I) according to claim 13, or the salt of the compound of formula (I) according to claim 14, or the pharmaceutical composition according to claim 15 in the preparation of a medicament for the prevention and / or treatment of diseases or conditions associated with elevated Lp(a) plasma levels.

18. The use according to claim 17, characterized in that, The diseases or conditions associated with elevated Lp(a) plasma levels are cardiovascular diseases (CVD); the cardiovascular diseases (CVD) include, but are not limited to, atherosclerotic cardiovascular disease (ASCVD), coronary artery stenosis, coronary artery-related diseases, aortic stenosis, aortic stenosis-related diseases, heart failure, heart failure-related diseases, atrial fibrillation, and atrial fibrillation-related diseases; the ASCVD includes peripheral vascular disease, peripheral artery-related diseases, coronary heart disease, ischemic stroke, and ischemic stroke-related diseases.

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