Salt and crystal form of benzo-aza aromatic ring derivative and application of salt and crystal form in medicine

Abstract

The invention relates to a pharmaceutical succinate of a compound shown in a formula (I) or a stereoisomer thereof, a crystal form, a preparation method, a pharmaceutical composition of the pharmaceutical succinate, and an application of the pharmaceutical succinate in preparation of drugs for treating diseases related to complement factor B activity or expression quantity.

Description

A salt and crystal form of a benzazepine aromatic ring derivative and its application in medicine Technical Field

[0001] The present invention relates to the field of medicine, and in particular to a pharmaceutically acceptable succinate salt of a compound represented by general formula (I) or a stereoisomer thereof, a crystal form thereof, and a preparation method thereof, as well as a pharmaceutical composition thereof and its use in preparing a drug for treating diseases related to complement factor B activity or expression. Background Art

[0002] Complement factor B is a component of the alternative complement pathway and participates in both specific and nonspecific immune mechanisms. It contains a serine protease (SP) domain that, when activated, provides catalytic activity for the C3 and C5 convertases of the alternative pathway. Complement factor B is a key enzyme in the activation of the alternative complement pathway and serves as a suitable target for inhibiting complement activation.

[0003] Patent PCT / CN2022 / 113216 describes a compound represented by formula (I), which exhibits excellent complement factor B inhibitory activity and a good inhibitory rate of C3a levels in vivo. When used as a therapeutic agent for human treatment, administering a therapeutically effective dose is problematic and can lead to toxic side effects or ineffective treatment. Therefore, it is crucial to select a salt form that is stable, reproducible, and possesses physicochemical properties favorable for its use as a therapeutic agent.

[0004] Summary of the Invention

[0005] The present invention aims to provide a pharmaceutically acceptable succinate salt, crystal form and preparation method of a compound represented by formula (I) or a stereoisomer thereof, as well as its application in the preparation of a drug for treating diseases related to complement factor B activity or expression.

[0006] The advantages of the salts and crystalline forms of the compounds of the present invention include, but are not limited to, ease of processing and crystallization, convenient handling, ease of purification, ease of industrialization, good fluidity, ease of micronization, high solubility, low hygroscopicity, good pharmacokinetic properties and good stability, and are suitable for the preparation of pharmaceutical preparations.

[0007] The present invention provides a succinate salt and a crystal form of a compound represented by formula (I).

[0008] In some embodiments, the pharmaceutically acceptable salt is selected from succinate;

[0009] In some embodiments, the molar ratio of the compound represented by formula (I): succinic acid is about 1:0.5 to 1:2.5;

[0010] In some embodiments, the molar ratio of the compound of formula (I): succinic acid is about 1:0.75;

[0011] The present invention relates to a crystalline form 1 of a succinate salt of the compound represented by the above formula (I), which has an X-ray powder diffraction pattern using Cu-Kα radiation, and has characteristic diffraction peaks at the following 2θ positions: 7.77°±0.2°, 8.17°±0.2°, 14.61°±0.2°, 17.06°±0.2°, and 20.00°±0.2°;

[0012] In some embodiments, the crystalline form 1 of the succinate salt of the compound represented by the aforementioned formula (I) has an X-ray powder diffraction pattern using Cu-Kα radiation having characteristic diffraction peaks at the following 2θ positions: 7.77°±0.2°, 8.17°±0.2°, 9.27°±0.2°, 12.76°±0.2°, 14.61°±0.2°, 17.06°±0.2°, 20.00°±0.2°, and 22.48°±0.2°;

[0013] In some embodiments, the crystalline form 1 of the succinate salt of the compound represented by the aforementioned formula (I) has an X-ray powder diffraction pattern using Cu-Kα radiation having characteristic diffraction peaks at the following 2θ positions: 7.34°±0.2°, 7.77°±0.2°, 8.17°±0.2°, 9.27°±0.2°, 12.50°±0.2°, 12.76°±0.2°, 13.69°±0.2°, 14.61°±0.2°, 17.06°±0.2°, 19.76°±0.2°, 20.00°±0.2°, 21.52°±0.2°, 22.01°±0.2°, 22.48°±0.2°, 22.56°±0.2°, and 24.53°±0.2°;

[0014] In some embodiments, the crystalline form 1 of the succinate salt of the compound represented by the aforementioned formula (I) has an X-ray powder diffraction pattern as shown in FIG1 using Cu-Kα radiation;

[0015] In some embodiments, the differential scanning calorimetry (DSC) curve of the crystalline form 1 of the succinate salt of the compound represented by the aforementioned formula (I) shows that the peak temperatures of the crystalline form 1 are 192.4° C. and 203.4° C., respectively; its thermogravimetric analysis (TGA) curve shows that the weight loss is about 1.67% before 150° C.; its isothermal adsorption curve shows that the weight gain is about 1.15% due to moisture absorption at 80% RH; its differential scanning calorimetry curve, thermogravimetric analysis curve, and isothermal adsorption curve are shown in Figures 2-4;

[0016] In some embodiments, the crystal structure of the compound represented by the aforementioned formula (I) was successfully determined by the MicroED method. The unit cell parameters are shown in Table 1. The crystal structure belongs to the orthorhombic system, C2221 (No. 20) space group, and the unit cell constant is α=90°,β=90°,γ=90°,unit cell volume Z' of the system is 2; in some embodiments, the asymmetric unit of Form 1 consists of 2 molecules of the compound represented by Formula (I) and 1.5 molecules of succinic acid; in some embodiments, the asymmetric unit in the unit cell is shown in FIG5 .

[0017] Table 1 Unit cell parameters of the succinate salt crystal form 1 of the compound represented by formula (I):

[0018] The present invention also relates to a method for preparing a crystalline form 1 of a succinate salt of a compound represented by formula (I), wherein the method comprises: ① taking the compound represented by formula (I), adding a solvent (i), stirring at room temperature or stirring under heating, ② adding an aqueous solution of succinic acid, stirring, crystallizing, and drying to obtain the target crystalline form 1; in some embodiments, the solvent used is selected from C 1-6 Halogenated alkane solvents, C 2-6 Ester solvents, C 2-6 Nitrile solvents, C 2-6 Ether solvents, C 1-6 One or more mixed solvents in any proportion of alcohol solvents or water, preferably one or more selected from dichloromethane, 1,2-dichloroethane, acetonitrile, ethyl acetate, methanol, ethanol, isopropanol, propanol, ether, tetrahydrofuran or water;

[0019] In some embodiments, in the preparation method of the aforementioned crystalline form 1, the solvent (i) is selected from a mixed solvent of ethanol and water or ethanol; the volume (ml) of the solvent (i) is 1 to 100 times, preferably 1 to 50 times, and more preferably 2 to 8 times the weight (g) of the compound.

[0020] In one aspect, the present invention further provides a pharmaceutical composition, wherein the pharmaceutical composition contains a therapeutically effective amount of the succinate salt, crystal form and pharmaceutically acceptable carrier or excipient of the compound represented by the aforementioned formula (I).

[0021] As used herein, an "effective amount" or "therapeutically effective amount" refers to administering a sufficient amount of a compound disclosed herein to alleviate, to some extent, one or more symptoms of the disease or condition being treated (e.g., kidney disease). In some embodiments, the result is a reduction and / or alleviation of the signs, symptoms, or causes of the disease, or any other desired change in a biological system. For example, an "effective amount" for therapeutic use is the amount of a composition comprising a compound disclosed herein required to provide a clinically significant reduction in disease symptoms.

[0022] In another aspect, the present invention also provides the use of the pharmaceutically acceptable salt of the compound represented by formula (I) or the succinate salt, crystal form or the aforementioned pharmaceutical composition of the compound represented by formula (I) in the preparation of drugs for diseases related to kidney diseases.

[0023] The present invention relates to a kit, which may include a composition in single-dose or multi-dose form, wherein the kit comprises a pharmaceutically acceptable salt or co-crystal of the compound of the present invention, and the amount of the pharmaceutically acceptable salt or co-crystal of the compound of the present invention is the same as that in the above-mentioned pharmaceutical composition.

[0024] The crystal form of the compound represented by formula (I) of the present invention has 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 crystal form of the present invention can significantly reduce the filtration time during the preparation process, shorten the production cycle, and save costs. The crystal form of the present invention also has good light stability, thermal stability, and moisture stability, which can ensure the reliability of the crystal form during storage and transportation, thereby ensuring the safety of the preparation, and the crystal form does not need to be specially packaged to prevent the influence of light, temperature, and humidity, thereby reducing costs. The crystal form will not be degraded due to the influence of light, high temperature, and high humidity, thereby improving the safety of the preparation and the effectiveness after long-term storage. Patients taking the crystal form will not worry about the preparation producing photosensitivity reactions due to exposure to sunlight.

[0025] The crystalline form of the compound represented by formula (I) of the present invention has good chemical and physical stability, is easy to prepare, and is more suitable for the preparation of formulations. The crystalline form of the present invention has good fluidity, good compressibility, high bulk density, low hygroscopicity, and uniform particle size distribution.

[0026] The crystal form of the compound represented by formula (I) described in the present invention is suitable and convenient for large-scale preparation. The preparation prepared 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 the expressions such as “preferably, ..., its X-ray powder diffraction pattern has a characteristic diffraction peak at the following 2θ position” or “more preferably, ..., its X-ray powder diffraction pattern has a characteristic diffraction peak at the following 2θ position” described in the present invention mean that in addition to having a characteristic diffraction peak at the aforementioned 2θ position, there is also a characteristic diffraction peak at the aforementioned “following 2θ position”.

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

[0029] The crystalline 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 form of the present invention is not limited to the characteristic spectra that are exactly the same as the characteristic spectra described in the drawings disclosed in the present invention, such as XRD, DSC, and TGA. Any crystal form having characteristic spectra that are substantially the same or essentially the same as those described in the drawings falls within the scope of the present invention.

[0031] It is understood that, as is well known in the art 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 compound of the present invention has a DSC pattern with characteristic peak positions, having substantially the same properties as the DSC pattern provided in the accompanying drawings of the present invention, with an error tolerance of ±3°C.

[0032] Unless otherwise stated, the technology and scientific terms used herein have the same meaning as those skilled in the art to which the present invention pertains. If there is a contradiction, the definition provided herein shall prevail. When a certain amount, concentration or other value or parameter is expressed in the form of a range, a preferred range or a preferred upper numerical limit and a preferred lower numerical limit, it should be understood that it is equivalent to specifically disclosing any scope by combining any pair of upper range limits or preferred values ​​with any lower range limit or preferred values, without considering whether the scope is specifically disclosed. Unless otherwise stated, the numerical ranges listed herein are intended to include all integers and fractions (decimals) within the endpoints and range of the range.

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

[0034] The term "optional" or "optionally" as used herein means that the subsequently described event or circumstance may but need not occur, and the description includes instances where the event or circumstance occurs or does not occur.

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

[0036] Unless otherwise indicated, percentages, parts, etc. herein are by weight.

[0037] As used herein, "amorphous" refers to any solid material that is not ordered in three dimensions. In some cases, amorphous solids can be characterized by known techniques, including XRPD crystal diffraction analysis, differential scanning calorimetry (DSC), solid-state nuclear magnetic resonance (ssNMR) spectroscopy, or a combination of these techniques. As described below, the XRPD pattern produced by an amorphous solid has no obvious characteristic diffraction peaks. Other characterization methods, such as differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), dynamic vapor phase adsorption (DVS), etc., further help to identify the crystalline form and help determine stability and solvent / water content.

[0038] As used herein, a "crystalline form" or "crystal" refers to any solid material that exhibits a three-dimensional ordering, as opposed to an amorphous solid material, which produces a characteristic XRPD pattern with well-defined peaks.

[0039] The "pharmaceutical composition" of the present invention refers to a mixture of one or more compounds described herein or their physiologically / pharmaceutically acceptable salts and other components, wherein the other components include physiologically / pharmaceutically acceptable carriers and excipients.

[0040] The "carrier" mentioned in the present invention refers to a carrier or diluent that does not cause significant irritation to the organism and does not eliminate the biological activity and properties of the administered compound.

[0041] As used herein, "excipient" refers to an inert substance added to a pharmaceutical composition to facilitate administration of the compound. 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, and the like.

[0042] The "ether solvent" described in the present invention refers to a chain compound or a cyclic compound 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.

[0043] The "alcohol solvent" described in the present invention refers to one or more "hydroxyl" substituted "C 1-6 The group is derived from one or more hydrogen atoms on the "alkyl" group, the "hydroxyl" and "C 1-6 "Alkyl" is as defined above, and specific examples include but are not limited to methanol, ethanol, isopropanol, n-propanol, isopentanol or trifluoroethanol.

[0044] The "ester solvent" mentioned in the present 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.

[0045] The "ketone solvent" described in the present invention refers to a compound in which a carbonyl group (-C(O)-) is connected to two hydrocarbon groups. Depending on the different hydrocarbon groups in the molecule, ketones can be divided 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.

[0046] The "nitrile solvent" of the present invention refers to one or more "cyano" substituted "C 1-6 The group derived from one or more hydrogen atoms on the "alkyl" group, the "cyano" and "C 1-6 "Alkyl" is as defined above, and specific examples include but are not limited to acetonitrile or propionitrile.

[0047] The "halogenated hydrocarbon solvent" of the present invention refers to one or more "halogen atoms" replacing "C 1-6 The group derived from one or more hydrogen atoms on the "alkyl", the "halogen atom" and "C 1-6 "Alkyl" is as defined above, and specific examples include but are not limited to: dichloromethane, 1,2-dichloroethane, chloroform or carbon tetrachloride.

[0048] The "room temperature" mentioned in the present invention generally refers to 4-30°C, preferably 20±5°C.

[0049] The drying temperature of the present invention is generally 20-100° C., preferably 25-70° C., and can be dried under normal pressure or reduced pressure (vacuum drying). Preferably, the drying is carried out under reduced pressure.

[0050] As used herein, an "X-ray powder diffraction pattern (XRPD pattern)" refers to an experimentally observed diffraction pattern or a parameter, data, or value derived therefrom. An XRPD pattern is typically characterized by peak positions (on the abscissa) and / or peak intensities (on the ordinate).

[0051] The term "2θ or 2θ angle" as used herein refers to the diffraction angle, where θ is the Bragg angle, which is a peak position expressed in degrees (°) based on an X-ray diffraction experiment, and is typically the horizontal axis unit in a diffraction pattern. If the incident beam is diffracted when the incident beam forms an angle θ with a certain lattice plane, the experimental setup requires recording the reflected beam at an angle of 2θ. It should be understood that the specific 2θ value of a specific 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 is ±0.3, which may be ±0.3, ±0.2, or ±0.1.

[0052] As used herein, "substantially the same" means that variations in representative peak positions and intensities are taken into account. For example, one skilled in the art will appreciate that peak positions (2θ) can exhibit some variation, typically as much as 0.1 to 0.2 degrees, and that the instrument used to measure diffraction can also introduce some variation. Furthermore, one skilled in the art will appreciate that relative peak intensities can vary due to instrumental differences, as well as the degree of crystallinity, preferred orientation, the surface of the sample being prepared, and other factors known to one skilled in the art, and should be considered merely qualitative measurements.

[0053] The "differential scanning calorimetry or DSC" mentioned in the present invention refers to measuring the temperature difference and heat flow difference between a sample and a reference object during the process of heating or maintaining the sample at a constant temperature, so as to characterize all physical and chemical changes related to thermal effects and obtain phase change information of the sample.

[0054] According to the description of hygroscopic characteristics and the definition of hygroscopic weight gain in the "9103 Guiding Principles for Hygroscopicity of Drugs" in Part IV of the 2020 edition of the Chinese Pharmacopoeia,

[0055] Deliquescent: Absorbs sufficient water to form a liquid;

[0056] Highly hygroscopic: weight gain due to moisture absorption is not less than 15%;

[0057] Hygroscopic: weight gain due to moisture absorption is less than 15% but not less than 2%;

[0058] Slightly hygroscopic: weight gain due to moisture absorption is less than 2% but not less than 0.2%;

[0059] No or almost no hygroscopicity: weight gain due to moisture is less than 0.2%.

[0060] The crystallization method of the present invention includes but is not limited to slurry crystallization, volatile crystallization, antisolvent crystallization, cooling crystallization, water vapor stress crystallization or diffusion crystallization.

[0061] The crystal form disclosed in the present invention can be prepared by the following common methods for preparing crystal forms:

[0062] 1. The volatilization experiment is to evaporate the clear sample solution at different temperatures until the solvent is dry.

[0063] 2. The slurry experiment is to stir the supersaturated solution of the sample (with insoluble solids present) at a certain temperature in different solvent systems.

[0064] 3. The antisolvent test is to dissolve the sample in a good solvent, add an antisolvent, stir the precipitated solid for a short time and then filter it immediately.

[0065] 4. The cooling crystallization experiment is to dissolve a certain amount of sample into the corresponding solvent at high temperature, and then stir and crystallize directly at room temperature or low temperature.

[0066] 5. The polymer template experiment is to add different types of polymer materials to the clear sample solution and leave it open at room temperature to evaporate until the solvent is dry.

[0067] 6. The thermal method experiment is to treat the sample according to certain thermal method crystallization conditions and cool it to room temperature.

[0068] 7. The water vapor diffusion experiment is to place the sample in a certain humidity environment at room temperature. BRIEF DESCRIPTION OF THE DRAWINGS

[0069] FIG1 is an X-ray powder diffraction pattern of Form 1 of the succinate salt of Compound 1.

[0070] FIG2 is a differential scanning calorimetry analysis curve of Form 1 of the succinate salt of Compound 1.

[0071] FIG3 is a thermogravimetric analysis spectrum of Form 1 of the succinate salt of Compound 1.

[0072] FIG4 is an isothermal adsorption curve of Form 1 of the succinate salt of Compound 1.

[0073] FIG5 is a diagram showing the single crystal structure of Form 1 of the succinate salt of Compound 1. DETAILED DESCRIPTION

[0074] The structures of the compounds were determined by nuclear magnetic resonance (NMR) or / and mass spectrometry (MS). NMR shifts (δ) were expressed in 10 -6 The unit of ppm is given. NMR measurements were performed using a WNMR-I 400 nuclear magnetic spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), deuterated methanol (CD3OD), and tetramethylsilane (TMS) as the internal standard.

[0075] XRD analysis was performed using a Malvern Panalytical X-ray powder diffractometer. The 2θ scanning angle ranged from 4° to 40°, with a scan step size of 0.01°. The tube voltage and current were 40 kV and 40 mA, respectively, and the sample pan was a zero-background sample pan.

[0076] TGA test conditions: Thermogravimetric analyzer (DSC 214) was used. A 1-10 mg sample was placed in a pre-equilibrated sample pan and automatically weighed in the TGA oven. The sample was heated at a rate of 10°C / min to the final temperature, with nitrogen purge at 30 mL / min.

[0077] DSC test conditions: Differential Scanning Calorimetry (DSC) was performed using a NETZSCH TG 209F3 differential scanning calorimeter. A 0.5-5 mg sample was accurately weighed and placed in a standard pan or perforated aluminum crucible sample pan. The sample was heated at a rate of 10°C / min to the final temperature. A nitrogen purge rate of 40 mL / min was used.

[0078] The known starting materials of the present invention can be synthesized by methods known in the art.

[0079] Unless otherwise specified in the examples, the solution refers to an aqueous solution.

[0080] The following describes in detail the implementation process of the present invention and the beneficial effects produced by specific embodiments, which is intended to help readers better understand the essence and characteristics of the present invention and is not intended to limit the scope of implementation of this case.

[0081] Example 1: Preparation of Compound 1

[0082] Step 1: Synthesis of intermediate B

[0083] tert-Butyl 4-(((2S,4R)-4-cyclopropyl-2-(4-(methoxycarbonyl)phenyl)piperidin-1-yl)methyl)-5-(methoxy-d3)-7-methyl-1H-indole-1-carboxylate (Compound B)

[0084] Compound A (162 mg, 0.625 mmol) was dissolved in 5 mL of isopropyl acetate, and maleic acid (73 mg, 0.628 mmol) was added. The reaction mixture was allowed to react at room temperature for 1 h. The reaction system was concentrated under reduced pressure to obtain a crude product (235 mg). This crude product (235 mg) was dissolved in 10 mL of ethanol, and tert-butyl 4-formyl-5-trideuteromethoxy-7-methyl-1H-indole-1-carboxylate (165 mg, 0.564 mmol) was added. 20 mg of Ir(CO)2acac was added, and the hydrogen atmosphere was replaced three times. The temperature was raised to 80°C, and the mixture was reacted under a hydrogen balloon for 16 h. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The crude product was then purified by silica gel column chromatography (petroleum ether / ethyl acetate (v / v) = 10:1) to obtain Compound B (200 mg, yield: 60%).

[0085] LCMS m / z=536.3[M+1] +

[0086] Step 2: Synthesis of compound 1

[0087] 4-((2S,4R)-4-cyclopropyl-1-((5-(methoxy-d3)-7-methyl-1H-indol-4-yl)methyl)piperidin-2-yl)benzoic acid (Compound 1)

[0088] Compound B (200 mg, 0.37 mmol) was dissolved in 5 mL of methanol, and solid potassium carbonate (257 mg, 1.86 mmol) was added. The mixture was heated to 80°C and refluxed for 3 h. The reaction solution was cooled to room temperature and concentrated under reduced pressure to obtain a crude product. The crude product was dissolved in a mixture of 5 mL of THF and 1 mL of water, and lithium hydroxide monohydrate (155 mg, 3.7 mmol) was added. The mixture was stirred at room temperature for 16 h. The reaction system was concentrated under reduced pressure, and the crude product was subjected to Pre-HPLC (Instrument and Preparative Column: Glison GX-281 Preparative HPLC, Sunfire C18 Preparative Column, 5 μm, ID x Length = 30 mm x 150 mm). Preparation Method: The crude product was dissolved in methanol and dimethyl sulfoxide and filtered through a 0.45 μm filter membrane to prepare a sample solution. The mobile phase system was acetonitrile / water (containing 0.1% TFA). Gradient elution method: acetonitrile was gradient eluted from 5% to 60% (elution time 15 min), and lyophilized to obtain the trifluoroacetic acid salt of compound 1 (125 mg).

[0089] 1H NMR (400MHz, CD3OD) δ8.28-8.18(m,2H),7.77-7.68(m,2H),7.36-7.30(m,1H),6.77( s,1H),6.32(d,1H),4.48(dd,1H),4.40-4.29(m,1H),4.18-4.08(m,1H),3.62-3.52(m ,1H),3.30-3.21(m,1H),2.51(s,3H),2.25-2.12(m,1H),2.09-1.85(m,2H),1.79-1.6 0(m,1H),1.20-1.03(m,1H),0.67-0.53(m,1H),0.52-0.40(m,2H),0.26-0.12(m,2H).

[0090] LCMS m / z=422.2[M+1] +

[0091] To 125 mg of trifluoroacetate of compound 1 were added 10 mL of 2-methyltetrahydrofuran and 50 mL of a saturated sodium bicarbonate solution, stirred for 15 min, extracted, and the organic phase was concentrated to dryness to obtain compound 1 (60 mg).

[0092] Example 2: Preparation of Form 1 of the succinate salt of Compound 1

[0093] Take about 250 mg of compound 1, add 7.5 mL of ethanol and 2.5 mL of water, stir at 25°C, add 15 mL of 47 mg / mL succinic acid aqueous solution, stir for 2 hours, filter, rinse the filter cake with 30% ethanol aqueous solution, and vacuum dry to obtain Form 1 of the succinate salt of compound 1.

[0094] 1. Crystal form test example

[0095] Table 2. Instrument information and detection method parameters

[0096] 2. Specific peak characterization results of XRD test of compound 1

[0097] The X-ray powder diffraction pattern (XRD) of the crystalline form of the succinate salt of Compound I is shown in Figure 1. The specific peaks are shown in Table 3.

[0098] Table 3

[0099] 3. Stability results

[0100] The stability results of the succinate salt form 1 of Compound 1 are shown in Table 4.

[0101] Table 4

[0102] Conclusion: The succinate salt form 1 of compound 1 showed good stability under accelerated, intermediate and long-term conditions.

[0103] 4. Pharmacokinetic Test in Rats

[0104] Experimental purpose: This study administered the test substance to SD rats by single-dose intravenous and oral gavage, determined the concentration of the test substance in rat plasma, and evaluated the pharmacokinetic characteristics of the test substance in rats.

[0105] Experimental animals: Male SD rats, 200-250 g, 6-8 weeks old, 6 rats / compound, purchased from Chengdu Dashuo Experimental Animal Co., Ltd.

[0106] Experimental method: On the day of the experiment, 6 SD rats were randomly divided into groups according to body weight. They were fasted but not watered for 12-14 hours before administration and fed 4 hours after administration.

[0107] Table 5 *Dosage is based on free base.

[0108] Sampling: Before and after drug administration, 0.15 mL of blood was collected from the eye socket under isoflurane anesthesia and placed in an EDTAK2 centrifuge tube. ℃ The mixture was centrifuged for 10 min and the plasma was collected.

[0109] The time points for plasma collection in the IV&PO group were: 0, 5 min, 15 min, 30 min, 1, 2, 4, 6, 8, and 24 h.

[0110] All samples were stored at -60°C before analysis and quantitative analysis was performed using LC-MS / MS.

[0111] Table 6 Pharmacokinetic parameters of the compounds of the present invention in rat plasma Test compounds *Note: Compounds were administered ig (orally)

[0112] Conclusion: The compounds of the present invention have good oral absorption in rats. Compared with LNP023, the trifluoroacetate salt of compound 1 has a higher maximum blood concentration and oral bioavailability in rats.

[0113] Reference compound (LNP023) structure:

[0114] 5. Canine Pharmacokinetic Testing

[0115] Experimental purpose: This study administered the test substance to beagle dogs by single-dose intravenous and oral administration, determined the concentration of the test substance in beagle dog plasma, and evaluated the pharmacokinetic characteristics of the test substance in beagle dogs.

[0116] Test animals: Male beagle dogs, 8-11 kg, 1-3 years old, 6 per compound, purchased from Beijing Masi Biotechnology Co., Ltd.

[0117] Experimental method: On the day of the experiment, 6 beagle dogs were randomly divided into groups according to body weight. They were fasted but not watered for 14-18 hours before administration and fed 4 hours after administration.

[0118] Table 7 *Dosage is based on free base.

[0119] Sampling: Before and after administration, collect 1.0 mL of blood from the jugular vein or limb vein and place it in an EDTAK2 centrifuge tube. Centrifuge at 5000 rpm, 4°C for 10 min to collect plasma.

[0120] The time points for plasma collection in the IV&PO group were: 0, 5 min, 15 min, 30 min, 1, 2, 4, 6, 8, 10, 12, and 24 h.

[0121] All samples were stored at -60°C before analysis and quantitative analysis was performed using LC-MS / MS.

[0122] Table 8 Pharmacokinetic parameters of the compounds of the present invention in dog plasma *Note: Compounds were administered ig (orally)

[0123] Conclusion: The compound of the present invention has good oral absorption in beagle dogs.

[0124] 6. Monkey Pharmacokinetic Test

[0125] Purpose: This study administered the test substance to cynomolgus monkeys by single-dose intravenous and oral administration, determined the concentration of the test substance in monkey plasma, and evaluated the pharmacokinetic characteristics of the test substance in monkeys.

[0126] Experimental animals: Male cynomolgus monkeys, 3-5 kg, 3-6 years old, 6 per compound, purchased from Suzhou Xishan Biotechnology Co., Ltd.

[0127] Experimental method: On the day of the experiment, 6 monkeys were randomly divided into groups according to body weight. They were fasted but not watered for 14-18 hours before administration and fed 4 hours after administration.

[0128] Table 9 *Dosage is based on free base.

[0129] Sampling: Before and after administration, 1.0 mL of blood was collected from the limb vein and placed in an EDTAK2 centrifuge tube. Centrifuge at 5000 rpm, 4°C for 10 min to collect plasma.

[0130] The time points for plasma collection in the IV&PO group were: 0, 5 min, 15 min, 30 min, 1, 2, 4, 6, 8, 10, 12, and 24 h.

[0131] All samples were stored at -60°C before analysis and quantitative analysis was performed using LC-MS / MS.

[0132] Table 10 Pharmacokinetic parameters of the compounds of the present invention in monkey plasma *Note: Compounds were administered ig (orally)

[0133] Conclusion: The compounds of the present invention have good oral absorption in monkeys. Compared with the trifluoroacetate salt of LNP023, the trifluoroacetate salt of compound 1 has higher oral exposure and bioavailability in monkeys.

[0134] 7. Caco2 permeability test

[0135] The experiment used Caco-2 cell monolayers in 96-well Transwell plates, incubated in triplicate. Transport buffer (HBSS, 10 mM HEPES, pH 7.4 ± 0.05) containing the compound of the invention (2 μM) or the control compounds digoxin (10 μM), nadolol (2 μM), and metoprolol (2 μM) was added to the apical or basolateral wells of the cell monolayer. Transport buffer containing DMSO was added to the corresponding receiving wells. After incubation at 37 ± 1°C for 2 hours, the cell plate was removed and appropriate samples were taken from the apical and basolateral wells to a new 96-well plate. Proteins were then precipitated by adding acetonitrile containing an internal standard. Samples were analyzed using LC-MS / MS to determine the concentrations of the compound of the invention and the control compound. The concentration data were used to calculate the apparent permeability coefficients for transport from the apical to basolateral side of the cell monolayer and from the basolateral to apical side, thereby calculating the efflux rate. The integrity of the cell monolayer after 2 hours of incubation was assessed by leakage of Lucifer Yellow.

[0136] Table 11 Caco2 test results of the compounds of the present invention

[0137] Conclusion: The compounds of the present invention have good Caco2 permeability. Compared with the trifluoroacetate salt of LNP023, the trifluoroacetate salt of compound 1 has better permeability and lower efflux rate.

Claims

1. A pharmaceutically acceptable salt of the compound represented by formula (I), The pharmaceutically acceptable salt is selected from succinate, wherein: The molar ratio of the compound represented by formula (I): succinic acid is about 1:0.

75.

2. A crystalline form 1 of a succinate salt of a compound represented by formula (I), wherein the crystalline form 1 is an orthorhombic system, a C2221 (No. 20) space group, and a unit cell constant of α=90°,β=90°,γ=90°,unit cell volume The Z' of the system is 2.

3. A crystalline form 1 of the succinate salt of the compound represented by formula (I), using Cu-Kα radiation, and its X-ray powder diffraction pattern has characteristic diffraction peaks at the following 2θ positions (2θ±0.2°): 7.77°±0.2°, 8.17°±0.2°, 14.61°±0.2°, 17.06°±0.2°, and 20.00°±0.2°.

4. The crystalline form 1 of the succinate salt of the compound represented by formula (I) according to claim 2, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks at the following 2θ positions (2θ±0.2°): 7.77°±0.2°, 8.17°±0.2°, 9.27°±0.2°, 12.76°±0.2°, 14.61°±0.2°, 17.06°±0.2°, 20.00°±0.2°, and 22.48°±0.2°.

5. The crystalline form 1 of the succinate of the compound represented by formula (I) according to claim 2, using Cu-Kα radiation, has an X-ray powder diffraction pattern having characteristic diffraction peaks at the following 2θ positions (2θ±0.2°): 7.34°±0.2°, 7.77°±0.2°, 8.17°±0.2°, 9.27°±0.2°, 12.50°±0.2°, 12.76°±0.2°, 13.69°±0.2°, 14.61°±0.2°, 17.06°±0.2°, 19.76°±0.2°, 20.00°±0.2°, 21.52°±0.2°, 22.01°±0.2°, 22.48°±0.2°, 22.56°±0.2°, and 24.53°±0.2°.

6. The crystal form 1 of the succinate according to claim 2, using Cu-Kα radiation, has an X-ray powder diffraction pattern as shown in FIG1 .

7. The crystal form 1 of the succinate according to claim 2, whose differential scanning calorimetry analysis curve and thermogravimetric analysis curve are shown in Figure 2 and Figure 3 respectively.

8. A method for preparing the crystalline form 1 of the succinate salt of the compound represented by formula (I), wherein: The method comprises: ① taking a compound represented by formula (I), adding a solvent (i), stirring at room temperature or heating and stirring, ② adding a succinic acid aqueous solution, stirring, crystallizing, and drying to obtain the target crystal form 1.

9. The preparation method according to claim 8, wherein: The solvent used is selected from C 1-6 Halogenated alkane solvents, C 2-6 Ester solvents, C 2-6 Nitrile solvents, C 2-6 Ether solvents, C 1-6 One or more mixed solvents in any proportion of alcohol solvents or water are preferably selected from one or more of dichloromethane, 1,2-dichloroethane, acetonitrile, ethyl acetate, methanol, ethanol, isopropanol, propanol, ether, tetrahydrofuran or water.

10. A pharmaceutical composition, wherein The pharmaceutical composition contains a therapeutically effective amount of the pharmaceutically acceptable salt according to claim 1, or the crystal form according to any one of claims 2 to 7, and a pharmaceutically acceptable carrier or excipient.

11. The pharmaceutical composition according to claim 10, wherein the therapeutically effective amount is 1-800 mg calculated as the free base.

12. Use of the pharmaceutically acceptable salt according to claim 1, or the crystal form according to claims 2 to 7, or the pharmaceutical composition according to claim 10 in the preparation of a medicament for treating a disease associated with complement factor B activity or expression.

13. The use according to claim 12, wherein the disease is selected from kidney disease.

Images