Crystal form of bet protein inhibitor, medical composition, preparation method and use

Abstract

The invention relates to the technical field of pharmaceutical preparations, in particular to a BET protein inhibitor crystal form, a pharmaceutical composition, a preparation method and application. The pharmaceutical composition comprises an active ingredient of a compound as shown in a formula (I), the pharmaceutical composition has excellent dissolution of the active ingredient and excellent absorbability in vivo. # imgabs0 #

Description

Technical Field

[0001] The present application relates to the technical field of pharmaceutical preparations, and in particular to a crystal form, pharmaceutical composition, preparation method and use of a BET protein inhibitor. Background Art

[0002] Histone acetylation is an important part of epigenetic research. Acetylated histones can activate gene transcription through the action of DNA polymerase, RNA polymerase and transcription factors. The bromodomain and extra-terminal domain (BET) family belongs to the bromodomain protein family (BRDs), which is a class of evolutionarily highly conserved proteins that can recognize and bind to acetylated lysine residues in the tail of histones, recruit chromatin regulatory proteins, transcription factors, chromatin remodeling factors, etc., and thus play an important role in regulating gene transcription and chromatin remodeling. It is related to various biological processes such as cell growth, proliferation and differentiation, apoptosis and necrosis, and is an important epigenetic "reader".

[0003] So far, it has been found that the human genome encodes a total of 61 bromodomains, which are distributed in 46 different proteins. The BRDs family consists of four subtypes: BRD2, BRD3, BRD4, and BRDT, all of which contain two tandem bromodomains (BD1, BD2) and an extra-terminal (ET) domain. The two bromodomains are mainly responsible for recognizing and binding to acetylated lysine residues, while the ET domain interacts with auxiliary factors. Abnormal expression of BET proteins is associated with a variety of diseases. Although the four members of the BET family have similar structures, their biological functions are still different, especially BRD4 is closely related to a variety of diseases such as cancer and inflammation (Jung M et al., Epigenomics, 2015, 7(3): 487-501.).

[0004] Targeting BET proteins is beneficial for developing new therapeutic strategies targeting cancer, inflammation and viruses. Currently, small molecule inhibitors targeting BET proteins have entered the preclinical and clinical research stages, and are mainly used to treat cancer and autoimmune diseases. For example: JQ1, a BET small molecule inhibitor developed by Picaud et al. (Picaud S et al., Nature, 2010, 468(7327): 1067-1073.) in 2010. Related biological experiments have shown that JQ1 has potential therapeutic value for diseases such as cancer, cardiovascular disease, human immunodeficiency virus (HIV) infection and inflammation; RVX-208 developed by Resverlogix Corp. (McLure KG et al., PLoS One, 2013, 8, 12: e83190.) is a quinazolinone inhibitor that selectively acts on BD2. Clinical data show that it has significant therapeutic effects on diseases such as coronary artery syndrome, atherosclerosis and diabetes, and has now entered Phase III clinical trials; Incyte disclosed compound INCB-057643 in patent application CN106414442A, which is in Phase I clinical trials; Abbive disclosed a class of selective BET protein inhibitors in WO2017177955A1, among which compound ABBV-744 showed better safety in preclinical studies, but is currently in Phase I clinical trials and has been suspended.

[0005] Therefore, there is a need to develop pharmaceutical compositions of BET protein inhibitors. Summary of the invention

[0006] The purpose of the present application is to provide a crystal form, a pharmaceutical composition, a preparation method and a use of a BET protein inhibitor; the active ingredient in the pharmaceutical composition has good solubility and dissolution, and excellent absorption in the body.

[0007] The present application provides a pharmaceutical composition, which comprises an active ingredient, wherein the active ingredient is a compound represented by formula (I);

[0008]

[0009] In some embodiments, the pharmaceutical composition contains 2-100 mg, preferably 5-50 mg, and more preferably 5-15 mg, of the active ingredient per unit preparation, for example 2, 10, 15, 25, 30, 40, 50, 60, 70, 80, 90, 100 mg.

[0010] In some embodiments, the active ingredient is 1.5% to 25% of the total weight of the pharmaceutical composition, preferably 1.5% to 20%, more preferably 3.7% to 5.6%, for example: 1.5%, 3.7%, 5.6%, 9.3%, 12%, 16.7%, 20%, 25%.

[0011] In some embodiments, the pharmaceutical composition further comprises: one or more pharmaceutically acceptable excipients;

[0012] Preferably, the excipient is selected from one or more of a filler, a binder, a disintegrant, a solubilizer and a lubricant;

[0013] More preferably, the auxiliary materials include: fillers, binders, disintegrants, solubilizers and lubricants.

[0014] In some embodiments, the filler is selected from one or more of microcrystalline cellulose, lactose, cellulose-lactose, starch, mannitol, and calcium hydrogen phosphate, preferably cellulose-lactose. In the present invention, cellulose-lactose is a mixture of lactose and powdered cellulose, for example, comprising a mixture containing 75% by weight of lactose and 25% by weight of powdered cellulose. Cellulose-lactose can use cellulose-lactose Cellactose 80, but is not limited thereto.

[0015] In some embodiments, the binder is selected from one or more of povidone, hydroxypropyl methylcellulose, hydroxypropyl cellulose, and polyvinyl alcohol, preferably povidone, for example, povidone K30.

[0016] In some embodiments, the disintegrant is selected from one or more of cross-linked sodium carboxymethyl cellulose, cross-linked polyvinylpyrrolidone, sodium carboxymethyl starch, and low-substituted hydroxypropyl cellulose, preferably cross-linked sodium carboxymethyl cellulose.

[0017] The term "low-substituted hydroxypropyl cellulose" refers to low-substituted hydroxypropyl cellulose in which the content of hydroxypropyl groups in the hydroxypropyl cellulose is 5.0 wt% to 9.9 wt%.

[0018] In some embodiments, the solubilizing agent is selected from one or more of sodium lauryl sulfate, Tween, Span, polyoxyethylene fatty acid esters, preferably sodium lauryl sulfate.

[0019] In some embodiments, the lubricant is selected from one or more of magnesium stearate, calcium stearate, and sodium stearyl fumarate, preferably magnesium stearate.

[0020] In some embodiments, each of the pharmaceutical compositions comprises: 3 to 19 mg of the binder, preferably 3 to 12 mg, more preferably 3 to 6 mg, for example: 3, 6, 10, 12, 16, 19 mg.

[0021] In some embodiments, the pharmaceutical composition comprises: 20-318 mg of the filler, preferably 66-193 mg, more preferably 97-193 mg, for example: 20, 58, 63, 66, 76, 82, 105, 107, 115, 116, 119, 121, 124, 153, 170, 193, 213, 250, 280, 300, 318 mg.

[0022] In some embodiments, the pharmaceutical composition comprises: 1 to 300 mg of the solubilizer, preferably 1 to 45 mg, more preferably 5 to 45 mg, for example: 1, 6, 10, 15, 30, 45, 50, 75, 100, 150, 180, 200, 250, 280, 300 mg.

[0023] In some embodiments, the pharmaceutical composition comprises: 1 to 25 mg of the disintegrant, preferably 4 to 28 mg, more preferably 4 to 8 mg, for example: 1, 3, 4, 5, 7, 8, 9, 15, 20, 25 mg.

[0024] In some embodiments, the pharmaceutical composition includes: 0.7-10 mg of the lubricant, preferably 0.7-5 mg, preferably 0.7-2.7 mg, for example: 0.7, 1, 1.4, 1.8, 2, 2.5, 2.7, 3, 4, 5, 8, 10 mg.

[0025] In some embodiments, the pharmaceutical composition comprises: each unit of the pharmaceutical composition comprises:

[0026] The active ingredient is 2 to 100 mg, preferably 5 to 50 mg, more preferably 5 to 15 mg;

[0027] The binder is 3 to 19 mg, preferably 3 to 12 mg, more preferably 3 to 6 mg;

[0028] The filler is 20 to 318 mg, preferably 66 to 193 mg, more preferably 97 to 193 mg;

[0029] The solubilizer is 1 to 300 mg, preferably 1 to 45 mg, more preferably 5 to 45 mg;

[0030] The disintegrant is 1 to 25 mg, preferably 4 to 28 mg, more preferably 4 to 8 mg;

[0031] The lubricant is 0.7 to 10 mg, preferably 0.7 to 5 mg, and more preferably 0.7 to 2.7 mg.

[0032] In some embodiments, the pharmaceutical composition comprises: each unit of the pharmaceutical composition comprises:

[0033] 2 to 100 parts of the active ingredient, preferably 5 to 50 parts, more preferably 5 to 15 parts;

[0034] The binder is 3 to 19 parts, preferably 3 to 12 parts, more preferably 3 to 6 parts;

[0035] The filler is 20 to 318 parts, preferably 66 to 193 parts, more preferably 97 to 193 parts;

[0036] The solubilizer is 1 to 300 parts, preferably 1 to 45 parts, more preferably 5 to 45 parts;

[0037] The disintegrant is 1 to 25 parts, preferably 4 to 28 parts, more preferably 4 to 8 parts;

[0038] The lubricant is present in an amount of 0.7 to 10 parts, preferably 0.7 to 5 parts, and more preferably 0.7 to 2.7 parts.

[0039] In this application, parts are in milligrams.

[0040] In some embodiments, the binder is added by internal addition, the filler is added by internal addition or internal and external addition, and the disintegrant, solubilizer and lubricant are added by external addition.

[0041] In some embodiments, the filler is added by internal and external addition, and the filler is added in two parts, one part is the filler of the internal addition part added inside the particle, and the other part is the filler of the external addition part added outside the particle. In some embodiments, the filler of the internal addition part and the filler of the external addition part can be the same or different. In some embodiments, the filler of the internal addition part is the same as the filler of the external addition part.

[0042] In some embodiments, the weight ratio of the internal filler to the external filler is (32-64): (1-254).

[0043] In some embodiments, the pharmaceutical composition includes: 32-64 mg of an added portion of filler, for example: 32, 40 mg, 50 mg, 64 mg.

[0044] In some embodiments, the pharmaceutical composition includes: an additional portion of filler 1 to 254 mg, preferably 75 to 129 mg, for example: 1, 2, 6, 12, 18, 50, 64, 73, 74, 75, 76, 82, 85, 86, 87, 89, 91, 102, 129, 149, 200, 220, 254 mg.

[0045] In some embodiments, the filler is 4% to 92% of the total weight of the pharmaceutical composition, preferably 72% to 86%; for example: 92%, 90%, 88%, 87%, 86%, 85%, 84%, 82%, 80%, 79%, 78%, 72%, 70%, 64%, 60%, 57%, 50%, 40%, 30%, 27%, 16%, 14%, 12%, 10%, 5%, 4%.

[0046] In some embodiments, the binder is 2% to 12% of the total weight of the pharmaceutical composition, preferably 2.2%; for example: 2%, 2.2%, 2.4%, 5%, 8%, 10%, 12%.

[0047] In some embodiments, the disintegrant is 1% to 5% of the total weight of the pharmaceutical composition, preferably 3%; for example: 1%, 2%, 3%, 4%, 5%.

[0048] In some embodiments, the solubilizer is 0.7% to 60% of the total weight of the pharmaceutical composition, preferably 3.7% to 18.5%; and / or 0.7%, 3.7%, 4.4%, 7.4%, 10%, 11.1%, 16.7%, 18.5%, 27.8%, 40%, 50%, 60%.

[0049] In some embodiments, the weight ratio of the compound represented by Formula I to the solubilizer is 1:0.5-5, preferably 1:1-5, such as 1:0.5, 1:1, 1:3, 1:5.

[0050] In some embodiments, the lubricant is 0.5% to 2.0% of the total weight of the pharmaceutical composition, for example: 0.5%, 1.0%, 2.0%.

[0051] In some embodiments, the pharmaceutical composition is an oral preparation, preferably an oral solid preparation; more preferably a tablet, granule or capsule, further preferably a tablet or capsule.

[0052] In the present application, the granules are granulated into a granular preparation. In addition, as for the granules, a coating can be added as needed, and the granules can be made into enteric granules or sustained-release granules using a known appropriate method. The granules can be foaming granules, which refer to granules that rapidly foam and dissolve or disperse in water.

[0053] In the present application, the tablets may be manufactured by tableting. For example, the mixed raw materials may be directly tableted to obtain tablets, or the mixed raw materials may be made into granules and further compressed to obtain tablets.

[0054] In the present application, in the case of capsules, they may be hard capsules or soft capsules.

[0055] In the present application, a capsule can be obtained by filling the granules into a capsule.

[0056] In the present application, the compound represented by formula (I) may include a hydrate of the compound represented by formula (I). The hydrate may be prepared using a known method. Specifically, the hydrate may be a compound represented by formula (II);

[0057]

[0058] Among them, n=0-6, for example: n=0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5 or 6.

[0059] In the present application, the compound represented by formula (I) may be a polymorph. In some specific embodiments, the compound represented by formula (I) may be a crystalline form I, and the X-ray powder diffraction pattern of the crystalline form I has characteristic peaks at 2θ angles of 4.65±0.20°, 7.19±0.20°, 8.31±0.20°, 8.77±0.20°, 10.13±0.20°, 15.19±0.20°, 18.49±0.20° and 20.28±0.20°.

[0060] The present invention provides a crystalline form I of a compound represented by formula (I), wherein an X-ray powder diffraction spectrum has characteristic peaks at 2θ angles of 4.65±0.20°, 7.19±0.20°, 8.31±0.20°, 8.77±0.20°, 10.13±0.20°, 15.19±0.20°, 18.49±0.20° and 20.28±0.20°.

[0061] In some embodiments, the X-ray powder diffraction pattern of the crystalline Form I has characteristic peaks at 2θ angles of 4.18±0.2°, 4.65±0.2°, 5.09±0.2°, 7.19±0.2°, 8.31±0.2°, 8.77±0.2°, 10.13±0.2°, 13.06±0.2°, 13.86±0.2°, 14.66±0.2°, 15.19±0.2°, 15.65±0.2°, 17.54±0.2°, 17.69±0.2°, 18.49±0.2°, 20.28±0.2°, 20.94±0.2° and 25.92±0.2°.

[0062] In some embodiments, the X-ray powder diffraction pattern of the crystalline form I at a 2θ angle is 4.18±0.2°, 4.65±0.2°, 5.09±0.2°, 7.19±0.2°, 8.31±0.2°, 8.77±0.2°, 10.13±0.2°, 10.78±0.2°, 11.33±0.2°, 11.70±0.2°, 13.06±0.2°, There are characteristic peaks at .2°, 13.56±0.2°, 13.86±0.2°, 14.66±0.2°, 15.19±0.2°, 15.65±0.2°, 17.54±0.2°, 17.69±0.2°, 18.49±0.2°, 20.28±0.2°, 20.66±0.2°, 20.94±0.2° and 25.92±0.2°.

[0063] In some embodiments, the X-ray powder diffraction pattern of the crystalline form I at a 2θ angle of 4.18±0.2°, 4.65±0.2°, 5.09±0.2°, 7.19±0.2°, 8.31±0.2°, 8.77±0.2°, 10.13±0.2°, 10.78±0.2°, 11.33±0.2°, 11.70±0.2°, 13.06±0.2°, 13.56±0.2°, 1 There are characteristic peaks at 3.86±0.2°, 14.66±0.2°, 15.19±0.2°, 15.65±0.2°, 17.54±0.2°, 17.69±0.2°, 18.49±0.2°, 20.28±0.2°, 20.66±0.2°, 20.94±0.2°, 23.99±0.2°, 25.92±0.2°, 26.37±0.2° and 26.69±0.2°.

[0064] In some embodiments, the X-ray powder diffraction pattern of Form I is substantially as follows Figure 1 shown.

[0065] In some embodiments, the TGA spectrum of the crystalline form I is substantially as follows Figure 2 shown.

[0066] In some embodiments, the DSC spectrum of the crystalline form I has endothermic peaks at 102.86°C±5°C, 139.19°C±5°C, and 218.03°C±5°C.

[0067] In some embodiments, the DSC spectrum of the crystalline form I is substantially as follows Figure 3 shown.

[0068] The present invention also provides a method for preparing the above-mentioned pharmaceutical composition, comprising the step of mixing the active ingredient and one or more pharmaceutically acceptable excipients.

[0069] In some embodiments, the step of mixing the active ingredient, filler, binder, disintegrant, solubilizer and lubricant is included;

[0070] In some embodiments, the granules are prepared by dry granulation, wet granulation, fluidized bed granulation or direct mixing; preferably, the solvent used in wet granulation or fluidized bed granulation is alcohol; preferably, the prepared granules can be further prepared into tablets, or directly filled into capsules to prepare capsules.

[0071] In some embodiments, the preparation method comprises:

[0072] 1) Preparation of drug-loaded solution;

[0073] 2) Preparation of drug-loaded particles;

[0074] 3) Preparation of total mixed particles;

[0075] 4) Optionally, capsule filling or tableting.

[0076] In some embodiments, the preparation method comprises:

[0077] 1) mixing a solvent, a binder and a compound of formula I to prepare a drug-loaded solution;

[0078] 2) mixing the added filler with the drug-loaded solution, granulating, and drying to obtain drug-loaded particles;

[0079] 3) mixing the drug-loaded particles, the additional filler, the solubilizer, the disintegrant and the lubricant to prepare the total mixed particles;

[0080] 4) Optionally, capsule filling or tableting; or

[0081] 1) mixing a solvent, a binder and a compound of formula I to prepare a drug-loaded solution;

[0082] 2) mixing the filler with the drug-loaded solution, granulating, and drying to obtain drug-loaded particles;

[0083] 3) mixing the drug-loaded particles, filler, solubilizer, disintegrant and lubricant to prepare total mixed particles;

[0084] 4) Optionally, capsule filling or tableting;

[0085] As used herein, unless otherwise stated, the term "optional" means that the object or event it modifies may or may not exist, or may or may not occur.

[0086] More preferably, the solvent is ethanol, and the weight ratio of the binder to the ethanol is 6 to 1:42, preferably 1:42;

[0087] More preferably, the step 2) adopts fluidized bed granulation.

[0088] In some embodiments, the drying temperature is 40-60°C.

[0089] In some specific embodiments, the preparation method comprises:

[0090] 1) Preparation of drug-loaded solution:

[0091] Take ethanol, add povidone under stirring until it is completely dissolved, add the active ingredient, and stir until it is completely dissolved to prepare a drug-loaded solution;

[0092] 2) Preparation of drug-loaded particles:

[0093] The fluidized bed is preheated for 3 minutes with an inlet air temperature of 30-40°C, and then the drug-loaded solution is slowly sprayed in, and the material temperature is controlled at 25-35°C. After the spraying is completed, the material is dried with an inlet air temperature of 40-60°C, and the moisture content of the material is controlled to be ≤3%. After the moisture content is qualified, the material temperature is lowered to below 40°C and the material is discharged to obtain drug-loaded particles.

[0094] 3) Preparation of total mixed particles:

[0095] The drug-loaded particles, an additional portion of cellulose-lactose, sodium lauryl sulfate, cross-linked sodium carboxymethyl cellulose and magnesium stearate are mixed to prepare total mixed particles.

[0096] 4) Capsule filling or tableting.

[0097] In some specific embodiments, the preparation method comprises:

[0098] 1) Preparation of drug-loaded solution:

[0099] Take ethanol, add povidone under stirring until it is completely dissolved, add the active ingredient, stir until it is completely dissolved, and prepare a drug-loaded solution.

[0100] 2) Preparation of drug-loaded particles:

[0101] After adding part of the cellulose-lactose into the fluidized bed and preheating it with an inlet air temperature of 30-40°C for about 3 minutes, slowly spray the drug-loaded solution and control the material temperature at 25-35°C. After the spraying is completed, use an inlet air temperature of about 40-60°C to dry the material and control the moisture content of the material to ≤3%. After the moisture content is qualified, reduce the material temperature to below 40°C and discharge the material.

[0102] 3) Preparation of total mixed particles:

[0103] The drug-loaded particles, an additional portion of cellulose-lactose, sodium lauryl sulfate, cross-linked sodium carboxymethyl cellulose and magnesium stearate are mixed to obtain total mixed particles.

[0104] 4) Capsule filling or tableting.

[0105] The present invention also provides the use of the above-mentioned pharmaceutical composition, the crystal form of the compound represented by the above-mentioned formula (I) or the pharmaceutical composition prepared by the above-mentioned preparation method in preparing drugs for preventing and / or treating diseases related to BET proteins;

[0106] In some embodiments, the BET protein-related diseases include, but are not limited to, tumor diseases, inflammatory diseases, autoimmune diseases, and viral infections.

[0107] In some embodiments, the tumor disease includes but is not limited to non-solid tumors (such as leukemia), solid tumors (cancers and sarcomas), such as acute leukemia, acute lymphocytic leukemia, acute myeloid leukemia (including monocytic leukemia, myeloblastic leukemia, myelomonocytic leukemia and promyelocytic leukemia), acute T-cell leukemia, B-cell acute lymphoid leukemia, angiosarcoma, astrocytoma, basal cell carcinoma, bile duct cancer, bladder cancer, brain cancer, breast cancer, bronchial cancer, prostate cancer, cervical cancer, chondrosarcoma, chordoma, choriocarcinoma, chronic leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia and chronic myeloid leukemia, chronic myeloid leukemia, colon cancer, colorectal cancer, non-small cell lung cancer.

[0108] In some specific embodiments, the BET protein-related disease is a hematological malignancy.

[0109] The BET protein inhibitor of the present application having a structure as shown in the compound of formula (I) has good BET protein inhibitory activity and good pharmacokinetic properties, and can also exhibit good in vivo tumor inhibition activity and low risk of hERG cardiotoxicity, and is a new generation of highly effective and low-toxic BET protein inhibitors.

[0110] The present application also covers solutions obtained by any combination, deletion or replacement of the above-mentioned embodiments and preferred embodiments.

[0111] Patent application PCT / CN2021 / 134454 (application date 2021.11.30) studies a class of BET protein inhibitors, among which the compound shown in formula (I) prepared in Example 1 shows excellent BRD4 inhibitory effect and is a potential new drug for diseases related to BET proteins.

[0112] BET bromodomain inhibitors have great application prospects as drug development, and there is a continuous clinical demand for the development of novel pharmaceutical compositions of highly effective and low-toxic BET protein inhibitors.

[0113] Beneficial effects of this application:

[0114] The pharmaceutical composition of the present application has high solubility, achieving the technical effect of improving relative bioavailability. Moreover, the pharmaceutical composition of the present application is stable even if stored for a long time.

[0115] The preparation method of the pharmaceutical composition of the present invention can significantly improve the in vivo absorption of the pharmaceutical composition and increase the relative bioavailability. BRIEF DESCRIPTION OF THE DRAWINGS

[0116] Figure 1 is the XRD pattern of Form I in Preparation Example 1;

[0117] Figure 2 is the TGA graph of Form I in Preparation Example 1;

[0118] Figure 3 This is the DSC graph of Form I in Preparation Example 1. DETAILED DESCRIPTION

[0119] In order to make the present application easier to understand, the present application will be described in detail below in conjunction with embodiments. These embodiments are only for illustration and are not limited to the scope of application of the present application.

[0120] Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0121] In this application, the terms "include", "comprises", "contains" and their equivalents should be understood as open, non-exclusive meanings, i.e., "including but not limited to", meaning that in addition to the listed elements, components and steps, other unspecified elements, components and steps may also be included. In this document, unless the context clearly dictates otherwise, singular terms encompass plural referents and vice versa.

[0122] The term "pharmaceutically acceptable" means that a carrier, vehicle, diluent, excipient, and / or formed salt is generally chemically or physically compatible with the other ingredients constituting a pharmaceutical dosage form and physiologically compatible with the receptor without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0123] The term "treatment" means administering the compound or formulation described herein to improve or eliminate a disease or one or more symptoms associated with the disease, and includes inhibiting the progression of a disease or condition, and alleviating a disease or condition.

[0124] In this application, all pharmaceutical excipients or reagents mentioned may be from commercial sources.

[0125] In this application, unless otherwise specified, "%" in the examples of this application refers to mass percentage.

[0126] In the present application, unless otherwise specified, the reaction temperature is 20-30°C.

[0127] In the present application, the active ingredient has the structure shown in formula (I);

[0128]

[0129] The compound represented by formula (I) may include a hydrate of the compound represented by formula (I). The specific hydrate may be prepared using a known method. Specifically, the hydrate may be a compound represented by formula (II):

[0130]

[0131] Among them, n=0-6, for example: n=0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5 or 6.

[0132] When n=2, it is a dihydrate crystal form of N-ethyl-4-(5-(2-hydroxypropyl-2-yl)-3-(indolyl-1-yl)thiophene-2-yl)-6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridine-2-carboxamide.

[0133] The pharmaceutical composition of the present application may contain the crystalline form I of the compound represented by formula (I) as an active ingredient. The crystalline form I of the compound represented by formula (I) is the crystalline form I of N-ethyl-4-(5-(2-hydroxypropyl-2-yl)-3-(indolyl-1-yl)thiophene-2-yl)-6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridine-2-carboxamide dihydrate.

[0134] The active ingredient in the preparation examples, embodiments, comparative examples and test examples of the present application adopts crystal form I.

[0135] In a specific embodiment, the preparation method of the crystal form I comprises: reacting a compound represented by formula (V), a solvent A and water to obtain a compound represented by formula (III);

[0136]

[0137] In a specific embodiment, the volume ratio of solvent A to water is 10:1-2:1, preferably 3:1; solvent A is preferably acetonitrile; wherein the volume mass ratio of solvent A to the compound represented by formula (V) is preferably 7.5 ml / g, and the volume mass ratio of water to the compound represented by formula (V) is preferably 2.5 ml / g;

[0138] In a specific embodiment, the preparation method of the crystal form I further comprises: reacting the compound represented by formula (IV) and a methyl metal reagent in a solvent B, post-treating, concentrating, adding dichloromethane and 1,4-dioxane for crystallization, to obtain the compound represented by formula (V);

[0139]

[0140] In a specific embodiment, the volume mass ratio of the solvent B to the compound represented by formula (IV) is 20-30 ml / g, preferably 30 ml / g; the solvent B is preferably selected from dimethyl sulfoxide, N,N-dimethylacetamide, N-methylpyrrolidone, 1,4-dioxane, tetrahydrofuran, and the solvent B is preferably tetrahydrofuran;

[0141] In a specific embodiment, the solvent B is preferably tetrahydrofuran;

[0142] In a specific embodiment, the molar ratio of the compound represented by formula (IV) to the methyl metal reagent is 1:4.0-5.0, preferably 1:4.0; in some preferred embodiments, the methyl metal reagent is selected from methyl lithium, methyl magnesium bromide, methyl magnesium chloride, methyl magnesium iodide, methyl zinc or methyl aluminum, preferably methyl lithium.

[0143] In a specific embodiment, the volume ratio of dichloromethane to 1,4-dioxane is 8:1.5-2.5, preferably 4:1, wherein the volume mass ratio of dichloromethane to the compound represented by formula (IV) is 8 ml / g, and the volume mass ratio of 1,4-dioxane to the compound represented by formula (IV) is 2 ml / g.

[0144] In some preferred embodiments, the crystallization temperature of the crystallization process is 10-25°C.

[0145] In some preferred embodiments, the crystallization time of the crystallization process is 14 to 18 hours.

[0146] In some embodiments, the crystallization by adding dichloromethane and 1,4-dioxane is specifically as follows: add dichloromethane, heat to 35-45°C, stir to dissolve, add 1,4-dioxane, cool to 15-25°C, stir for 5-7h, and then cool to 10-20°C, stir for 9-11h.

[0147] In some embodiments, the post-treatment is adding saturated water with ammonium chloride, separating the liquids, extracting the aqueous phase with ethyl acetate, combining the organic phases, washing with saturated brine, adding anhydrous sodium sulfate, stirring, filtering by pressure, and washing the filter cake with ethyl acetate.

[0148] In some embodiments, the reaction mixture is concentrated under reduced pressure until no liquid flows out, dichloromethane is added and the reaction mixture is concentrated until no liquid flows out, and the concentration is continued.

[0149] XRD: X-ray powder diffraction

[0150] The X-ray powder diffraction (XRD) described in this application is measured using a Malvern Panalytical Empyrean X-ray powder diffractometer. The test is based on the Pharmacopoeia of the People's Republic of China 2020 Edition, Part IV General Rules: 0451 X-ray Diffraction Method: Method 2 Powder X-ray Diffraction Method.

[0151] In this article, "X-ray powder diffraction pattern is basically Figure 1 "shown" refers to the X-ray powder diffraction pattern and Figure 1 Substantially the same. The term "substantially the same" with respect to X-ray powder diffraction patterns is meant to take into account variations in representative peak positions and intensities.

[0152] TGA: Thermogravimetric analysis

[0153] The thermogravimetric analysis (TGA) described in this application is performed using a TGA / DSC2 / 1600 simultaneous thermal analyzer. The test is based on JY / T 0589.4-2020 General Rules for Thermal Analysis Methods Part 4: Thermogravimetric Method.

[0154] The error of TGA can be within about ±0.5 mass %. Figure 2 "shown" refers to the TGA spectrum and Figure 2 Substantially the same. The term "substantially the same" with respect to TGA patterns means that such error variations are taken into account.

[0155] DSC: Differential Scanning Calorimetry

[0156] The differential scanning calorimetry (DSC) described in this application is measured using a DSC1 / 700 differential scanning calorimeter. The test is based on JY / T 0589.3-2020 General Rules for Thermal Analysis Methods Part 3: Differential Scanning Calorimetry.

[0157] In this article, "DSC spectra are basically Figure 3 "shown" refers to the DSC spectrum and Figure 3 Substantially the same, the term "substantially the same" of the DSC spectrum means that the representative characteristic peak positions are taken into account.

[0158] In this application, the dissolution curve was determined using the dissolution and release determination method in General Chapter 0931 of Part IV of the 2020 edition of the Chinese Pharmacopoeia.

[0159] Dissolution test method:

[0160] The dissolution and release rate was determined according to the dissolution and release rate determination method (Chinese Pharmacopoeia 2020 Edition, Part IV, General Rules 0931, Method 2, Pulp Method).

[0161] Dissolution conditions:

[0162] Dissolution medium pH 4.5: Take 2.99g sodium acetate, add 14.0ml 2mol acetic acid, add water to 1000ml, shake well; then add 1.0g sodium dodecyl sulfate (SDS), heat to dissolve, mix well, and you have it.

[0163] Dissolution medium pH 6.8: Use 900 ml of pH 6.8 phosphate buffer (weigh 7.8 g of sodium dihydrogen phosphate dihydrate, 0.896 g of sodium hydroxide, add 1000 ml of water to dissolve, add 0.5 g of sodium dodecyl sulfate to dissolve, mix well, and obtain) as the dissolution medium, place the test sample in a sinker, rotate at 75 revolutions per minute, operate according to the law, and take samples after 45 minutes.

[0164] Test solution: Take the eluate, filter it, and take the filtrate as the test solution.

[0165] Reference substance solution: Take an appropriate amount of reference substance, weigh accurately, add solvent [methanol-acetonitrile (50:50) v / v] to dissolve and dilute to make a solution with about 11 μg of the main component per 1 ml.

[0166] Determination method: Inject the test solution and the reference solution into the liquid chromatograph respectively, record the chromatogram, and calculate the dissolution amount of each particle by the peak area according to the external standard method.

[0167] Preparation Example 1: Preparation of N-ethyl-4-(5-(2-hydroxypropyl-2-yl)-3-(indolyl-1-yl)thiophene-2-yl)-6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridine-2-carboxamide dihydrate crystalline form

[0168] (1) Preparation of crude compound represented by formula (V)

[0169]

[0170] Feeding: Add 26.7 kg of tetrahydrofuran and 1.00 kg of the compound represented by formula (IV) to the reaction kettle, replace with nitrogen twice. Heat to 30-35°C and stir to dissolve, cool to 10±5°C after dissolution, drop 4.69 kg of methyl lithium reagent (keep at 10±10°C), keep at 10-20°C and stir for 30±10 minutes after the addition is complete.

[0171] Post-treatment: After the reaction is completed, add 7.8 kg of saturated ammonium chloride solution (keep at 15 ± 5 ° C), slowly add 6.0 kg of drinking water to the reactor, stir for 1.0 ± 0.5 h, let stand for 0.5 h, and separate the liquids. The aqueous phase is extracted with 5.4 kg of ethyl acetate. Combine the organic phases and wash with 10.0 kg of saturated brine. Add 4.00 kg of anhydrous sodium sulfate to the organic phase and stir for 2.0 ± 0.5 h. Filter the organic phase with a filter press, and wash the kettle wall and filter cake with 2.7 kg of ethyl acetate.

[0172] Concentration: The organic phase was transferred to a reactor, concentrated under reduced pressure at 40±5°C until no liquid flowed out, 3.33 kg of dichloromethane was added and concentrated until no liquid flowed out, and the concentration was continued for 0.5-1 h.

[0173] Crystallization: Add 6.65 kg of dichloromethane, heat to 40±5°C, stir to dissolve, filter through a precision filter, add 1.66 kg of 1,4-dioxane, cool to 20±5°C, stir for 6±1h, then cool to 15±5°C, stir for 10±1h.

[0174] Filtration: Filter by suction, rinse the reactor with 2.00 kg of ethyl acetate and wash the filter cake. Put the filter cake into a vacuum drying oven and dry it under reduced pressure at 40±5°C and vacuum degree ≤-0.080 MPa to obtain a crude compound represented by formula (V).

[0175] (2) Preparation of the refined product of the compound represented by formula (V)

[0176]

[0177] Feeding: Add 6.65 kg of dichloromethane, 1.00 kg of the crude compound represented by formula (V), and 0.24 kg of methanol into the reaction kettle, and stir at 25±5°C to dissolve.

[0178] Decolorization: Add 0.10 kg of activated carbon and stir at 25±5℃ for 3 hours.

[0179] Filtration and water washing: Filter the reaction solution with a precision filter, add 2.66 kg of dichloromethane to rinse the reactor, press the filtrate into the precision filter, and wash the organic phase twice with 4.0 kg of purified water.

[0180] Crystallization: Add 2.57 kg of 1,4-dioxane to the organic phase and stir at 25±5°C for 10±1 h, then cool to 5±5°C and stir for 6±1 h.

[0181] Suction filtration: Suction filtration, rinse the reactor with 2.00 kg of ethyl acetate and wash the filter cake.

[0182] Drying: Spread the filter cake evenly on a ceramic plate, with each plate not exceeding 2.0 kg, and place it in a square vacuum drying oven, and dry under reduced pressure at 40±5°C and vacuum degree ≤-0.080MPa. Drying is considered complete when the weight loss is ≤0.5%, and the refined product of the compound represented by formula (V) is obtained.

[0183] (3) Preparation of the compound represented by formula (III)

[0184]

[0185] Feeding: Add 5.92 kg acetonitrile, 2.50 kg purified water and 1.0 kg refined product of the compound represented by formula (V) into the reaction kettle. Stir at 20±5°C for 20±2 h under nitrogen protection.

[0186] Filtration: Rinse the reactor and the filter cake with a mixture of acetonitrile and aqueous solution (add 0.30 kg of purified water to 0.60 kg of acetonitrile), and then rinse the reactor and the filter cake with 2.00 kg of ethyl acetate.

[0187] Drying: Place the filter cake in a vacuum drying oven and dry it under reduced pressure at 45±5°C and vacuum degree ≤-0.080MPa to obtain the finished product of the compound represented by formula (III).

[0188] The obtained finished product of the compound represented by formula (III) is N-ethyl-4-(5-(2-hydroxypropyl-2-yl)-3-(indolyl-1-yl)thiophene-2-yl)-6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridine-2-carboxamide dihydrate crystalline form, named as crystalline form I.

[0189] The obtained crystal form I was subjected to X-ray powder measurement using Cu-ka radiation, and its spectrum had the diffraction angle, interplanar spacing and relative intensity shown in Table 1:

[0190] Table 1 Diffraction angle, peak height, interplanar spacing, relative intensity, and area of ​​Form A

[0191]

[0192]

[0193] The error of the 2θ diffraction angle is ±0.20°.

[0194] Furthermore, the crystalline form I prepared in Preparation Example 1 has substantially Figure 1 The X-ray powder diffraction pattern is shown.

[0195] The thermogravimetric analysis (TGA) spectrum of the crystal form I is substantially as follows Figure 2As shown in the figure, it can be seen that the weight loss of the crystal form in the range of 30°C to 165°C is 7.0%.

[0196] The differential scanning calorimetry (DSC) analysis of the crystal form I is substantially as follows Figure 3 As shown in the figure, it can be seen that the crystal form has endothermic peaks at 102.86℃±5℃, 139.19℃±5℃ and 218.03℃±5℃.

[0197] Preparation Example 2

[0198] Step 1: Preparation of drug-loaded solution

[0199] Weigh the prescribed amount of ethanol, add the prescribed amount of povidone K30 under stirring until it is completely dissolved, slowly add the prescribed amount of active ingredient (crystalline form I prepared by the preparation method of reference preparation example 1), stir until it is completely dissolved, prepare the drug-loaded solution, and set aside.

[0200] Step 2: Preparation of drug-loaded particles

[0201] The cellulose-lactose added to the fluidized bed is preheated for about 3 minutes with an inlet air temperature of 30-40°C, and then the drug-loaded solution is slowly sprayed in, and the material temperature is controlled at 25-35°C. After the spraying is completed, the material is dried with an inlet air temperature of about 40-60°C, and the material moisture is controlled to be ≤3%. After the moisture is qualified, the material temperature is reduced to below 40°C and the material is discharged to obtain drug-loaded particles.

[0202] Step 3: Preparation of total mixed particles

[0203] The amount of the additional excipients is converted according to the content and weight of the drug-loaded particles, and the drug-loaded particles are mixed with the added cellulose-lactose, sodium lauryl sulfate, cross-linked sodium carboxymethyl cellulose and magnesium stearate to obtain total mixed particles.

[0204] Step 4: Capsule filling

[0205] The filling amount is calculated based on the total mixed granule content for filling or tableting.

[0206] Preparation Example 3

[0207] Step 1: Preparation of drug-loaded solution

[0208] Weigh the prescribed amount of ethanol, add the prescribed amount of povidone K30 under stirring until it is completely dissolved, slowly add the prescribed amount of active ingredient (crystalline form I prepared by the preparation method of reference preparation example 1), stir until it is completely dissolved, prepare the drug-loaded solution, and set aside.

[0209] Step 2: Preparation of drug-loaded particles

[0210] After adding cellulose-lactose to the fluidized bed and preheating it with an inlet air temperature of 30-40°C for about 3 minutes, the drug-loaded solution is slowly sprayed in, and the material temperature is controlled at 25-35°C. After the spraying is completed, the material is dried with an inlet air temperature of about 40-60°C, and the moisture content of the material is controlled to be ≤3%. After the moisture content is qualified, the material temperature is reduced to below 40°C and the material is discharged to obtain drug-loaded particles.

[0211] Step 3: Preparation of total mixed particles

[0212] The amount of the additional excipients is converted according to the content of the drug-loaded particles and the weight, and the drug-loaded particles are mixed with sodium lauryl sulfate, cross-linked sodium carboxymethyl cellulose and magnesium stearate to obtain total mixed particles.

[0213] Step 4: Capsule filling

[0214] The filling amount is calculated according to the total mixed particle content.

[0215] Example 1

[0216] Prepared by the preparation method of reference example 2.

[0217] Table 1 Capsule preparation / tablet (1000 capsules / tablet, each unit preparation contains 2 mg of active ingredient)

[0218]

[0219] Example 2

[0220] Prepared by the preparation method of reference example 2.

[0221] Table 2 Capsule preparation / tablet (1000 capsules / tablet, each unit preparation contains 100 mg of active ingredient)

[0222]

[0223]

[0224] Example 3

[0225] Table 3 Capsule preparation / tablet (1000 capsules / tablet, each unit preparation contains 5 mg of active ingredient)

[0226]

[0227] Example 4

[0228] Prepared by the preparation method of reference example 2.

[0229] Table 4 Capsule preparation (1000 capsules, each unit preparation contains 10 mg of active ingredient)

[0230]

[0231] Example 5

[0232] Prepared by the preparation method of reference example 2.

[0233] Table 5 Capsule preparation (1000 capsules, each unit preparation contains 15 mg of active ingredient)

[0234]

[0235]

[0236] Example 6

[0237] Prepared by the preparation method of reference example 2.

[0238] Table 6 Capsule preparation (1000 capsules, each unit preparation contains 25 mg of active ingredient)

[0239]

[0240] Example 7

[0241] Prepared by the preparation method of reference example 2.

[0242] Table 7 Capsule preparation (1000 capsules, each unit preparation contains 50 mg of active ingredient)

[0243]

[0244] Example 8

[0245] Prepared by the preparation method of reference example 2.

[0246] Table 8 Capsule preparation / tablet (1000 capsules / tablet, each unit preparation contains 100 mg of active ingredient)

[0247]

[0248] Example 9

[0249] Prepared by the preparation method of reference example 2.

[0250] Table 9 Capsule preparation (1000 capsules, each unit preparation contains 2 mg of active ingredient)

[0251]

[0252] Example 10

[0253] Prepared by the preparation method of reference example 3.

[0254] Table 10 Capsule preparation (1000 capsules, each unit preparation contains 100 mg of active ingredient)

[0255]

[0256] Embodiment 11

[0257] Prepared by the preparation method of reference example 2.

[0258] Table 11 Capsule preparation (1000 capsules, each unit preparation contains 2 mg of active ingredient)

[0259]

[0260] Example 12

[0261] Prepared by the preparation method of reference example 2.

[0262] Table 12 Capsule preparation (1000 capsules, each unit preparation contains 2 mg of active ingredient)

[0263]

[0264] Example 13

[0265] Prepared by the preparation method of reference example 2.

[0266] Table 13 Capsule preparation (1000 capsules, each unit preparation contains 100 mg of active ingredient)

[0267]

[0268]

[0269] Embodiment 14

[0270] Prepared by the preparation method of reference example 3.

[0271] Table 14 Capsule preparation (1000 capsules, each unit preparation contains 100 mg of active ingredient)

[0272]

[0273] Embodiment 15

[0274] Prepared by the preparation method of reference example 2.

[0275] Table 15 Capsule preparation (1000 capsules, each unit preparation contains 2 mg of active ingredient)

[0276]

[0277] Example 16

[0278] Prepared by the preparation method of reference example 2.

[0279] Table 16 Capsule preparation (1000 capsules, each unit preparation contains 2 mg of active ingredient)

[0280]

[0281]

[0282] Embodiment 17

[0283] Prepared by the preparation method of reference example 2.

[0284] Table 17 Capsule preparation (1000 capsules, each unit preparation contains 100 mg of active ingredient)

[0285]

[0286] Embodiment 18

[0287] Prepared by the preparation method of reference example 2.

[0288] Table 18 Capsule preparation (1000 capsules, each unit preparation contains 2 mg of active ingredient)

[0289]

[0290] Embodiment 19

[0291] Prepared by the preparation method of reference example 2.

[0292] Table 19 Capsule preparation (1000 capsules, each unit preparation contains 2 mg of active ingredient)

[0293]

[0294]

[0295] Embodiment 20

[0296] Prepared by the preparation method of reference example 2.

[0297] Table 20 Capsule preparation (1000 capsules, each unit preparation contains 100 mg of active ingredient)

[0298]

[0299] Embodiment 21

[0300] Prepared by the preparation method of reference example 3.

[0301] Table 21 Capsule preparation (1000 capsules, each unit preparation contains 100 mg of active ingredient)

[0302]

[0303] Embodiment 22

[0304] Prepared by the preparation method of reference example 2.

[0305] Table 22 Capsule preparation (1000 capsules, each unit preparation contains 5 mg of active ingredient)

[0306]

[0307]

[0308] Embodiments 23-24

[0309] Prepared by the preparation method of reference example 2.

[0310] Table 23 Capsule preparation (1000 capsules, each unit preparation contains 5 mg of active ingredient)

[0311]

[0312] Embodiments 25-26

[0313] Prepared by the preparation method of reference example 2.

[0314] Table 24 Capsule preparation (1000 capsules, each unit preparation contains 5 mg of active ingredient)

[0315]

[0316]

[0317] Examples 27-28

[0318] Prepared by the preparation method of reference example 2.

[0319] Table 25 Capsule preparation (1000 capsules, each unit preparation contains 5 mg of active ingredient)

[0320]

[0321] Comparative Example 1

[0322] Comparative Preparations: Prescribing Information

[0323] Table 26 Capsules (1000 capsules, each unit preparation contains 15 mg of active ingredient)

[0324]

[0325] Comparative preparation method:

[0326] Step 1: Granulation

[0327] The active ingredient (crystalline form I prepared by the preparation method of reference preparation example 1), povidone, lactose, microcrystalline cellulose, and added sodium starch glycolate are mixed, and purified water is added to perform granulation, granulation, drying, and dry granulation to obtain dry granules.

[0328] Step 2: Total Mixing

[0329] The amount of the additional auxiliary materials is converted according to the weight of the dry particles, and mixed with the added sodium carboxymethyl starch and magnesium stearate to obtain total mixed particles.

[0330] Step 3: Capsule filling

[0331] Calculate the filling amount according to the total mixed powder content and fill it.

[0332] Test Example 1 Influencing Factor Test

[0333] 1. Test materials

[0334] Samples prepared according to Example 3 (capsules, each unit preparation containing 5 mg of active ingredient).

[0335] 2. Experimental Methods

[0336] The influencing factor tests were carried out under high temperature (60℃), high temperature (50℃), high humidity (RH75%), and strong light (light 4500Lux±500Lux, near-ultraviolet energy 1w / m2) conditions to determine the properties, related substances, solubility, moisture and content.

[0337] 3. Test results

[0338] Table 27

[0339]

[0340] The test results show that after being placed in strong light for 12 days, there is no obvious change in the sample compared with the day 0; after being placed in high temperature (60℃, 50℃) for 30 days, there is no obvious change in the indicators compared with the day 0; after being placed in high humidity (RH75%) conditions for 30 days, there is no obvious change in the indicators compared with the day 0.

[0341] Test Example 2 Dissolution curve investigation: Prescriptions with different binder dosages

[0342] 1. Test materials

[0343] Samples prepared according to Example 3 (capsules, each unit preparation containing 5 mg of active ingredient) and Example 22 (capsules, each unit preparation containing 5 mg of active ingredient).

[0344] 2. Experimental Methods

[0345] Determine the dissolution curve of the samples.

[0346] 3. Test results

[0347] Table 28

[0348]

[0349]

[0350] The measurement results show that the samples prepared according to Example 3 and Example 22 have a cumulative dissolution amount greater than 85% in 15 minutes.

[0351] Experimental Example 3 Dissolution curve investigation: Prescriptions with different disintegrant dosages

[0352] 1. Test materials

[0353] Samples prepared according to Example 23 (capsules, each unit preparation containing 5 mg of active ingredient), Example 3 (capsules, each unit preparation containing 5 mg of active ingredient), and Example 24 (capsules, each unit preparation containing 5 mg of active ingredient).

[0354] 2. Experimental Methods

[0355] Determination of sample dissolution curve

[0356] 3. Test results

[0357] Table 29

[0358]

[0359] The measurement results show that the samples prepared according to Example 23, Example 3, and Example 24 all have a cumulative dissolution amount greater than 85% in 15 minutes.

[0360] Experimental Example 4 Dissolution Curve Investigation: Prescriptions with Different Solubilizer Doses

[0361] 1. Test materials

[0362] Samples were prepared according to Example 3, Example 25 (capsules, each unit preparation containing 5 mg of active ingredient) (capsules, each unit preparation containing 5 mg of active ingredient), and Example 26 (capsules, each unit preparation containing 5 mg of active ingredient).

[0363] 2. Experimental Methods

[0364] Determination of sample dissolution curve

[0365] 3. Test results

[0366] Table 30

[0367]

[0368]

[0369] The measurement results show that the samples prepared according to Example 3, Example 25 and Example 26 all have a cumulative dissolution amount greater than 85% in 15 minutes.

[0370] Experimental Example 5 Dissolution Curve Investigation: Prescriptions with Different Lubricant Doses

[0371] 1. Test materials

[0372] Samples prepared according to Example 3 (capsules, each unit preparation containing 5 mg of active ingredient), Example 27 (capsules, each unit preparation containing 5 mg of active ingredient), and Example 28 (capsules, each unit preparation containing 5 mg of active ingredient).

[0373] 2. Experimental Methods

[0374] Determination of sample dissolution curve

[0375] 3. Test results

[0376] Table 31

[0377]

[0378] The measurement results show that the samples prepared according to Example 3, Example 27 and Example 28 all have a cumulative dissolution amount greater than 85% in 15 minutes.

[0379] Test Example 6: Dissolution curve investigation

[0380] 1. Test materials

[0381] Comparative preparation sample: Comparative preparation sample prepared according to Comparative Example 1

[0382] Sample of the present invention: Sample prepared according to Example 5 (capsules, each unit preparation containing 15 mg of active ingredient)

[0383] 2. Experimental Methods

[0384] Determination of sample dissolution curve

[0385] 3. Test results

[0386] Table 32

[0387]

[0388]

[0389] Results: The dissolution of the sample of Example 5 was significantly higher than that of the comparative preparation.

[0390] Test Example 7: Bioavailability Study

[0391] 1. Test materials

[0392] 1.1 Drugs

[0393] Comparative drug sample: active ingredient (using crystal form I)

[0394] Comparative preparation sample: Comparative preparation sample prepared according to Comparative Example 1

[0395] Sample of the present invention: Sample prepared according to Example 5 (capsules, each unit preparation containing 15 mg of active ingredient)

[0396] 1.2 Test equipment

[0397] LC-MS / MS instrument model: Triple Quad4500, AB Sciex (USA

[0398] 1.3 Research subjects

[0399] Six Beagle dogs, half male and half female, weighing 8±2kg, were purchased from Chengdu Dashuo Biotechnology Co., Ltd.

[0400] 2. Test methods

[0401] Bioavailability study: After a single oral administration of the sample of Example 5 of the present invention, the comparative preparation sample and the active ingredient to Beagle dogs at the same dose, the concentration level of the active ingredient in plasma and its basic pharmacokinetic characteristics were determined, and the differences in the main parameters such as Cmax, Tmax, AUClast, etc. were compared to evaluate its bioavailability activity.

[0402] Blood collection points: before administration, 0.5h, 1, 2, 4, 6, 8, and 24h.

[0403] Blood sample processing and storage: 2 mL of blood was collected from the forelimb vein and placed in an EDTA-K2 anticoagulant tube. Centrifuge at 4500 r / min for 10 min, separate 50 ul of plasma into a centrifuge tube, and freeze at -80°C.

[0404] Sample analysis: Take 50 μL of plasma, add 150 μL of 5 ng / mL memantine acetonitrile solution, and mix thoroughly; centrifuge at 12000 rpm for 10 min, take 100 μL of supernatant into an EP tube, add 100 μL of 50:50 (methanol: water) v / v solution and mix well, take 5 μL for analysis by LC-MS / MS.

[0405] 3. Test results

[0406] Table 33

[0407]

[0408] Results: The maximum blood concentration (Cmax) of the active ingredient in the sample of Example 5 of the present invention was approximately 3.56 times and 2.20 times that of the comparative preparation and the control drug (active ingredient), respectively, and the area under the curve (AUC) was approximately 8.92 times and 6.26 times that of the comparative preparation and the control drug (active ingredient), respectively, and the relative bioavailability was significantly higher.

[0409] In summary, the preparation according to the prescription and process of the embodiment of this patent can significantly improve its in vivo absorption and increase its relative bioavailability.

[0410] For the purpose of description and disclosure, all patents, patent applications and other publications are expressly incorporated herein by reference. These publications are provided only because they are disclosed prior to the filing date of the present application. All statements about the dates of these documents or the representations of the contents of these documents are based on the information available to the applicant and do not constitute any admission of the correctness of the dates of these documents or the contents of these documents. Moreover, any reference to these publications in this document does not constitute an admission that the publications are part of the common general knowledge in the art in any country.

[0411] Those skilled in the art will recognize that the scope of the present application is not limited to the various specific implementations and examples described above, but that various modifications, replacements, or recombinations can be made without departing from the spirit of the present application, which all fall within the scope of protection of the present application.

Claims

1. A pharmaceutical composition, characterized in that It comprises an active ingredient, wherein the active ingredient is a compound represented by formula (I); 2. The composition according to claim 2, characterized in that The pharmaceutical composition contains 2 to 100 mg, preferably 5 to 50 mg, and more preferably 5 to 15 mg of active ingredient per unit preparation.

3. The composition according to claim 1 or 2, characterized in that The pharmaceutical composition further comprises: one or more pharmaceutically acceptable excipients; Preferably, the excipient is selected from one or more of a filler, a binder, a disintegrant, a solubilizer and a lubricant; More preferably, the auxiliary materials include: fillers, binders, disintegrants, solubilizers and lubricants.

4. The composition according to claim 3, characterized in that The filler is selected from one or more of microcrystalline cellulose, lactose, cellulose-lactose, starch, mannitol, and calcium hydrogen phosphate, preferably cellulose-lactose; and / or The binder is selected from one or more of povidone, hydroxypropyl methylcellulose, hydroxypropyl cellulose and polyvinyl alcohol, preferably povidone; and / or The disintegrant is selected from one or more of cross-linked sodium carboxymethyl cellulose, cross-linked polyvinylpyrrolidone, sodium carboxymethyl starch, and low-substituted hydroxypropyl cellulose, preferably cross-linked sodium carboxymethyl cellulose; and / or The solubilizing agent is selected from one or more of sodium lauryl sulfate, Tween, Span, polyoxyethylene fatty acid ester, preferably sodium lauryl sulfate; and / or The lubricant is selected from one or more of magnesium stearate, calcium stearate, and sodium stearyl fumarate, preferably magnesium stearate.

5. The composition according to claim 3 or 4, characterized in that The pharmaceutical composition comprises: 3 to 19 mg, preferably 3 to 12 mg, more preferably 3 to 6 mg of the binder; and / or The pharmaceutical composition comprises: 20 to 318 mg of the filler, preferably 66 to 193 mg, more preferably 97 to 193 mg; and / or The pharmaceutical composition comprises: 1 to 300 mg, preferably 1 to 45 mg, more preferably 5 to 45 mg of the solubilizer; and / or The pharmaceutical composition comprises: 1 to 25 mg, preferably 4 to 28 mg, more preferably 4 to 8 mg of the disintegrant; and / or The pharmaceutical composition comprises: 0.7 to 10 mg of the lubricant, more preferably 0.7 to 5 mg; Preferably, the pharmaceutical composition comprises: each unit of the pharmaceutical composition comprises: The active ingredient is 2 to 100 mg, preferably 5 to 50 mg, more preferably 5 to 15 mg; The binder is 3 to 19 mg, preferably 3 to 12 mg, more preferably 3 to 6 mg; The filler is 20 to 318 mg, preferably 66 to 193 mg, more preferably 97 to 193 mg; The solubilizing agent is 1 to 300 mg, preferably 1 to 45 mg, more preferably 5 to 45 mg; The disintegrant is 1 to 25 mg, preferably 4 to 28 mg, more preferably 4 to 8 mg; The lubricant is 0.7 to 10 mg, preferably 0.7 to 5 mg, and more preferably 0.7 to 2.7 mg.

6. The composition according to claim 3 or 4, characterized in that The pharmaceutical composition is an oral preparation, preferably an oral solid preparation; more preferably a tablet, granule or capsule, further preferably a tablet or capsule.

7. The composition according to claim 3 or 4, characterized in that The compound represented by formula (I) is a polymorph, preferably form I, and the X-ray powder diffraction pattern of form I has characteristic peaks at 2θ angles of 4.65±0.20°, 7.19±0.20°, 8.31±0.20°, 8.77±0.20°, 10.13±0.20°, 15.19±0.20°, 18.49±0.20° and 20.28±0.20°.

8. A crystalline form I of a compound represented by formula (I), characterized in that: Its X-ray powder diffraction pattern has characteristic peaks at 2θ angles of 4.65±0.20°, 7.19±0.20°, 8.31±0.20°, 8.77±0.20°, 10.13±0.20°, 15.19±0.20°, 18.49±0.20° and 20.28±0.20°; Preferably, the X-ray powder diffraction pattern of the crystalline form I has characteristic peaks at 2θ angles of 4.18±0.20°, 4.65±0.20°, 5.09±0.20°, 7.19±0.20°, 8.31±0.20°, 8.77±0.20°, 10.13±0.20°, 13.06±0.20°, 13.86±0.20°, 14.66±0.20°, 15.19±0.20°, 15.65±0.20°, 17.54±0.20°, 17.69±0.20°, 18.49±0.20°, 20.28±0.20°, 20.94±0.20° and 25.92±0.20°; Preferably, the X-ray powder diffraction pattern of the crystalline form I at a 2θ angle is 4.18±0.20°, 4.65±0.20°, 5.09±0.20°, 7.19±0.20°, 8.31±0.20°, 8.77±0.20°, 10.13±0.20°, 10.78±0.20°, 11.33±0.20°, 11.70±0.20°, 13.06±0.20° , 13.56±0.20°, 13.86±0.20°, 14.66±0.20°, 15.19±0.20°, 15.65±0.20°, 17.54±0.20°, 17.69±0.20°, 18.49±0.20°, 20.28±0.20°, 20.66±0.20°, 20.94±0.20°, and 25.92±0.20°, with characteristic peaks; Preferably, the X-ray powder diffraction pattern of the crystalline form I is substantially as shown in FIG1 ; More preferably, the TGA spectrum of the crystalline form I is substantially as shown in Figure 2; More preferably, the DSC spectrum of the crystal form I has endothermic peaks at 102.86℃±5℃, 139.19℃±5℃ and 218.03℃±5℃; further preferably, the DSC spectrum of the crystal form is substantially as shown in Figure 3.

9. A method for preparing the pharmaceutical composition according to any one of claims 1 to 7, characterized in that: The method comprises the steps of mixing the active ingredient and one or more pharmaceutically acceptable excipients; Preferably, the method comprises the step of mixing the active ingredient, filler, binder, disintegrant, solubilizer and lubricant; Preferably, the granules are prepared by dry granulation, wet granulation, fluidized bed granulation or direct mixing; preferably, the solvent C used in wet granulation or fluidized bed granulation is water, alcohol or a mixture of alcohol and water; preferably, the prepared granules can be further prepared into tablets, or directly filled into capsules to prepare capsules; Preferably, the preparation method comprises: 1) Preparation of drug-loaded solution; 2) Preparation of drug-loaded particles; 3) Preparation of total mixed particles; 4) Optionally, capsule filling or tableting; Preferably, the preparation method comprises: 1) mixing solvent C, a binder and a compound of formula I to prepare a drug-loaded solution; 2) mixing the added filler with the drug-loaded solution, granulating, and drying to obtain drug-loaded particles; 3) mixing the drug-loaded particles, the additional filler, the solubilizer, the disintegrant and the lubricant to prepare the total mixed particles; 4) Optionally, capsule filling or tableting; or 1) mixing solvent C, a binder and a compound of formula I to prepare a drug-loaded solution; 2) mixing the filler with the drug-loaded solution, granulating, and drying to obtain drug-loaded particles; 3) mixing the drug-loaded particles, filler, solubilizer, disintegrant and lubricant to prepare total mixed particles; 4) Optionally, capsule filling or tableting; More preferably, the solvent C is ethanol, and the weight ratio of the binder to the ethanol is 6 to 1:42, preferably 1:42; More preferably, the step 2) adopts fluidized bed granulation.

10. Use of the pharmaceutical composition according to any one of claims 1 to 7, the crystal form of the compound represented by formula (I) according to claim 8, or the pharmaceutical composition prepared by the preparation method according to claim 9 in the preparation of drugs for preventing and / or treating diseases related to BET proteins; Preferably, the BET protein-related disease is selected from tumor disease, inflammatory disease, autoimmune disease, and viral infection; Preferably, the tumor disease comprises non-solid tumors, solid tumors, such as acute leukemia, acute lymphocytic leukemia, acute myeloid leukemia (including monocytic leukemia, myeloblastic leukemia, myelomonocytic leukemia and promyelocytic leukemia), acute T-cell leukemia, B-cell acute lymphocytic leukemia, angiosarcoma, astrocytoma, basal cell carcinoma, bile duct cancer, bladder cancer, brain cancer, breast cancer, bronchial cancer, prostate cancer, cervical cancer, chondrosarcoma, chordoma, choriocarcinoma, chronic leukemia, chronic lymphocytic leukemia, chronic myeloid leukemia and chronic myeloid leukemia, chronic myeloid leukemia, colon cancer, colorectal cancer, non-small cell lung cancer; More preferably, the BET protein-related disease is hematological malignancy.

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