Pyridinone substituents or derivatives thereof, methods of making, uses and pharmaceutical compositions

Abstract

The application provides a kind of pyridinone substituent or its derivative and its preparation method, application and pharmaceutical composition, belongs to chemical medicine technical field.The pyridinone substituent or its derivative is the compound shown in formula I, its salt, its stereoisomer, its solvate, its hydrate, its prodrug.The pyridinone substituent or its derivative of the application has good inhibition effect on BRD4-BD1 protein, can be used as new BET inhibitor, for preventing and / or treating various diseases related to BET, such as cancer, etc., excellent effect, has good application prospect.

Description

Technical Field

[0001] This invention belongs to the field of chemical pharmaceutical technology, specifically relating to a class of pyridone substitutes or their derivatives, their preparation methods, applications, and pharmaceutical compositions. Background Technology

[0002] Proteins containing a family of bromodomains and out-of-terminal (BET) domains are epigenetic readers; they regulate gene transcription by binding acetylated histones through their bromodomains. The BET family comprises four members: BRD2, BRD3, BRD4, and BRDT, sharing two N-terminal bromodomains and an additional C-terminal domain, exhibiting high sequence conservation. All BET family members play a role in controlling or executing various aspects of the cell cycle and remain complexed with chromosomes during cell division—meaning they play a role in maintaining epigenetic memory. Their dysfunction plays a crucial role in a variety of human diseases.

[0003] Inhibiting protein-protein interactions between BET protein and acetylated histones has become a potential therapeutic target for human diseases, including virological, inflammatory, central nervous system disorders, and various cancers. Reported small-molecule BET inhibitors include ABBV-075, GSK282015, OTX-015, CPI-0610, BI894999, BMS-986158, INCB054329, and GS-5829. There is a need to develop other BET inhibitors with improved properties compared to existing BET inhibitors, such as improved activity, selectivity, safety, tolerability, pharmacokinetic properties, or pharmacodynamics. Summary of the Invention

[0004] The purpose of this invention is to provide a class of pyridone substitutes or their derivatives, their preparation methods, applications, and pharmaceutical compositions.

[0005] This invention provides compounds of Formula I, their salts, their stereoisomers, their solvates, their hydrates, and their prodrugs:

[0006]

[0007] in,

[0008] R1 is selected from hydrogen, hydroxyl group, C1-C6 alkyl group, and NR5R6;

[0009] n is an integer from 0 to 3;

[0010] Each R a R b Each alkyl group is independently selected from hydrogen, substituted or unsubstituted C1 to C6 alkyl groups;

[0011] R2 is selected from hydrogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy;

[0012] R3 and R4 are each independently selected from hydrogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, -NHR7, halogen, nitro, cyano, carboxyl; or R3 and R4 are connected to pyridine to form a cyclic ring.

[0013] R5 and R6 are each independently selected from hydrogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted 6-10 aryl, substituted or unsubstituted 6-10 heteroaryl; or R5 and R6 are connected to N atoms to form substituted or unsubstituted 4-10 heterocyclic alkyl groups.

[0014] R7 is selected from hydrogen, substituted or unsubstituted C1 to C6 alkyl groups;

[0015] X is selected from S or N; Y is selected from CR8 or NR8;

[0016] The two dashed lines connecting X and Y are either empty or a key depending on the choice between X and Y. When the dashed line is empty, it is a single key; when the dashed line is a key, it is a double key. And the two dashed lines cannot be empty or a key at the same time.

[0017] R8 is selected from hydrogen, substituted or unsubstituted C1 to C6 alkyl groups;

[0018] Each substituent of the aryl and heteroaryl groups is independently selected from -C(O)N(H)R9;

[0019] R9 is selected from hydrogen, hydroxyl, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy;

[0020] Each substituent of the heterocyclic alkyl group is independently selected from substituted or unsubstituted C1-C6 alkyl groups, -S(O)(O)R 10 -C(O)R 10 3- to 10-membered cycloalkyl groups and hydroxyl groups;

[0021] R 10 Selected from substituted or unsubstituted C1-C6 alkyl groups and substituted or unsubstituted C1-C6 alkoxy groups;

[0022] Each substituent of the alkyl and alkoxy groups is independently selected from halogen, hydroxyl, amino, nitro, carboxyl, cyano, and 3- to 10-membered cycloalkyl groups.

[0023] Furthermore, the compound is as shown in Formula II:

[0024]

[0025] in,

[0026] R1 is selected from hydrogen, hydroxyl group, C1-C6 alkyl group, and NR5R6;

[0027] n is an integer from 0 to 3;

[0028] Each R a R b Each alkyl group is independently selected from hydrogen, substituted or unsubstituted C1 to C6 alkyl groups;

[0029] R2 is selected from hydrogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy;

[0030] X1 is selected from N or CH;

[0031] R5 and R6 are each independently selected from hydrogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted 6-10 aryl, substituted or unsubstituted 6-10 heteroaryl; or R5 and R6 are connected to N atoms to form substituted or unsubstituted 4-10 heterocyclic alkyl groups.

[0032] X is selected from S or N; Y is selected from CR8 or NR8;

[0033] The two dashed lines connecting X and Y are either empty or a key depending on the choice between X and Y. When the dashed line is empty, it is a single key; when the dashed line is a key, it is a double key. And the two dashed lines cannot be empty or a key at the same time.

[0034] R8 is selected from hydrogen, substituted or unsubstituted C1 to C6 alkyl groups;

[0035] Each substituent of the aryl and heteroaryl groups is independently selected from -C(O)N(H)R9;

[0036] R9 is selected from hydrogen, hydroxyl, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy;

[0037] Each substituent of the heterocyclic alkyl group is independently selected from substituted or unsubstituted C1-C6 alkyl groups, -S(O)(O)R 10 -C(O)R 10 3- to 10-membered cycloalkyl groups and hydroxyl groups;

[0038] R 10 Selected from substituted or unsubstituted C1-C6 alkyl groups and substituted or unsubstituted C1-C6 alkoxy groups;

[0039] Each substituent of the alkyl and alkoxy groups is independently selected from halogen, hydroxyl, amino, nitro, carboxyl, cyano, and 3- to 10-membered cycloalkyl groups.

[0040] Furthermore,

[0041] R1 is selected from hydrogen, hydroxyl group, C1-C3 alkyl group, and NR5R6;

[0042] R5 is selected from hydrogen or C1-C3 alkyl; R6 is selected from substituted or unsubstituted phenyl; or R5 and R6 are connected to a N atom to form a substituted or unsubstituted group.

[0043] n is 0 or 1;

[0044] Each R a R b Each is independently selected from hydrogen and C1-C3 alkyl groups;

[0045] R2 is selected from hydrogen and C1-C3 alkyl groups;

[0046] X1 is selected from N or CH;

[0047] X is selected from S or N; Y is selected from CR8 or NR8;

[0048] The two dashed lines connecting X and Y are either empty or a key depending on the choice between X and Y. When the dashed line is empty, it is a single key; when the dashed line is a key, it is a double key. And the two dashed lines cannot be empty or a key at the same time.

[0049] R8 is selected from hydrogen and C1-C3 alkyl groups;

[0050] Each substituent of the phenyl group is independently selected from -C(O)N(H)R9;

[0051] R9 is selected from hydrogen, hydroxyl, substituted or unsubstituted C1-C3 alkyl, substituted or unsubstituted C1-C3 alkoxy;

[0052] The Each substituent is independently selected from substituted or unsubstituted C1-C3 alkyl groups, -S(O)(O)R 10 -C(O)R 10 3- to 6-membered cycloalkyl groups and hydroxyl groups;

[0053] R 10 Selected from substituted or unsubstituted C1-C3 alkyl groups;

[0054] Each substituent of the alkyl and alkoxy groups is independently selected from halogen, hydroxyl, amino, nitro, carboxyl, cyano, and 3- to 6-membered cycloalkyl groups;

[0055] Preferably,

[0056] Selected from

[0057] R8 is selected from hydrogen and C1-C3 alkyl groups.

[0058] Furthermore, the compound is as shown in Formula III:

[0059]

[0060] in,

[0061] R5 is selected from hydrogen or C1-C3 alkyl; R6 is selected from substituted or unsubstituted phenyl; or R5 and R6 are connected to a N atom to form a substituted or unsubstituted group.

[0062] R2 is selected from C1-C3 alkyl groups;

[0063] X1 is selected from N or CH;

[0064] X is selected from S or N; Y is selected from CR8 or NR8;

[0065] The two dashed lines connecting X and Y are either empty or a key depending on the choice between X and Y. When the dashed line is empty, it is a single key; when the dashed line is a key, it is a double key. And the two dashed lines cannot be empty or a key at the same time.

[0066] R8 is selected from hydrogen and C1-C3 alkyl groups;

[0067] Each substituent of the phenyl group is independently selected from -C(O)N(H)R9;

[0068] R9 is selected from hydrogen, hydroxyl, substituted or unsubstituted C1-C3 alkyl, substituted or unsubstituted C1-C3 alkoxy;

[0069] The Each substituent is independently selected from substituted or unsubstituted C1-C3 alkyl groups, -S(O)(O)R 10 -C(O)R 10 3- to 6-membered cycloalkyl groups and hydroxyl groups;

[0070] R 10 Selected from substituted or unsubstituted C1-C3 alkyl groups;

[0071] Each substituent of the alkyl and alkoxy groups is independently selected from halogen, hydroxyl, amino, nitro, carboxyl, cyano, and 3- to 6-membered cycloalkyl groups;

[0072] Preferably,

[0073] Selected from

[0074] R8 is selected from hydrogen and C1-C3 alkyl groups.

[0075] Furthermore, the compound is as shown in Formula IV:

[0076]

[0077] in,

[0078] X2 is selected from NR', CHR', and O;

[0079] R' is selected from hydrogen, substituted or unsubstituted C1-C3 alkyl groups, and -S(O)(O)R. 10 -C(O)R 10 3- to 6-membered cycloalkyl groups and hydroxyl groups;

[0080] R 10 Selected from substituted or unsubstituted C1-C3 alkyl groups;

[0081] R2 is selected from C1-C3 alkyl groups;

[0082] X1 is selected from N or CH;

[0083] X is selected from S or N; Y is selected from CR8 or NR8;

[0084] The two dashed lines connecting X and Y are either empty or a key depending on the choice between X and Y. When the dashed line is empty, it is a single key; when the dashed line is a key, it is a double key. And the two dashed lines cannot be empty or a key at the same time.

[0085] R8 is selected from hydrogen and C1-C3 alkyl groups;

[0086] Each substituent of the alkyl group is independently selected from halogen, hydroxyl, amino, nitro, carboxyl, cyano, and 3- to 6-membered cycloalkyl groups.

[0087] Furthermore,

[0088] R' is selected from hydrogen, -S(O)(O)R 10 C1-C3 alkyl groups 3- to 6-membered cycloalkyl groups and hydroxyl groups;

[0089] R 10 Selected from C1 to C3 alkyl groups;

[0090] Selected from

[0091] R8 is selected from hydrogen and C1-C3 alkyl groups.

[0092] Furthermore, the compound is as shown in formula V:

[0093]

[0094] in,

[0095] R1 is selected from hydroxyl groups;

[0096] R2 is selected from hydrogen and C1-C3 alkyl groups;

[0097] X1 is selected from N or CH;

[0098] X is selected from S or N; Y is selected from CR8 or NR8;

[0099] The two dashed lines connecting X and Y are either empty or a key depending on the choice between X and Y. When the dashed line is empty, it is a single key; when the dashed line is a key, it is a double key. And the two dashed lines cannot be empty or a key at the same time.

[0100] R8 is selected from hydrogen and C1-C3 alkyl groups;

[0101] Preferably,

[0102] Selected from R8 is selected from hydrogen and C1-C3 alkyl groups.

[0103] Furthermore, the compound is one of the following compounds:

[0104]

[0105]

[0106] The present invention also provides the use of the aforementioned compounds, their salts, their stereoisomers, their solvates, their hydrates, and their prodrugs in the preparation of BET inhibitors and / or in the preparation of medicaments for the prevention and / or treatment of BET-related diseases.

[0107] Preferably, the BET inhibitor is a BRD4 inhibitor;

[0108] And / or, the BET-related diseases are cancer, inflammatory diseases, autoimmune diseases, or viral infections;

[0109] More preferably, the BRD4 inhibitor is an inhibitor of the BRD4-BD1 protein;

[0110] And / or, the cancer is one or more of the following: lymphoma, leukemia, breast cancer, lung cancer, liver cancer, colon cancer, myeloma, glioma, pancreatic cancer, fibrosarcoma, cervical cancer, and bladder cancer.

[0111] The present invention also provides a drug preparation which is a formulation made of the aforementioned compound, its salt, its stereoisomer, its solvate, its hydrate, and its prodrug as active ingredients, plus pharmaceutically acceptable excipients or auxiliary ingredients.

[0112] Alternatively, a pharmaceutical composition comprising the aforementioned compound, its salt, its stereoisomer, its solvate, its hydrate, or its prodrug.

[0113] The compounds and derivatives provided in this invention can be named according to the IUPAC (International Union of Pure and Applied Chemistry) or CAS (Chemical Abstracts Service, Columbus, OH) nomenclature system.

[0114] Regarding the definition of terms used in this invention: Unless otherwise stated, the initial definitions provided for groups or terms herein apply to the groups or terms used throughout this specification; for terms not specifically defined herein, the meanings that a person skilled in the art would give them should be given based on the disclosure and context.

[0115] "Substitution" refers to the replacement of hydrogen atoms in a molecule by other different atoms or molecules.

[0116] The minimum and maximum carbon atom content in hydrocarbon groups are indicated by a prefix, for example, the prefix C. a ~C b Alkyl indicates any alkyl group containing "a" to "b" carbon atoms. Therefore, for example, "C1 to C6 alkyl" refers to an alkyl group containing 1 to 6 carbon atoms; "C1 to C6 alkoxy" refers to an alkoxy group containing 1 to 6 carbon atoms.

[0117] "Alkyl" refers to a saturated hydrocarbon chain with a specified number of carbon atoms. For example, C1-C6 alkyl refers to an alkyl group with 1 to 6 carbon atoms, that is, alkyl groups with 1, 2, 3, 4, 5, or 6 carbon atoms. Alkyl groups can be straight-chain or branched. Representative branched alkyl groups have one, two, or three branches. Alkyl groups include methyl, ethyl, propyl (n-propyl and isopropyl), butyl (n-butyl, isobutyl, and tert-butyl), pentyl (n-pentyl, isopentyl, and neopentyl), and hexyl, etc.

[0118] "Halogen" refers to fluorine, chlorine, bromine, or iodine.

[0119] "Cycloalkyl" refers to saturated or unsaturated all-carbon monocyclic or polycyclic (including fused, spiro, or bridged rings) that do not possess a conjugated π-electron system, such as, but not limited to: wait.

[0120] "Heterocyclic alkyl" refers to a cycloalkyl group in which at least one carbon atom on the ring is replaced by a heteroatom, which is O, N, or S, and is a saturated or unsaturated monocyclic or polycyclic (including fused, spiro, or bridged rings) that does not have a conjugated π-electron system, such as including but not limited to:

[0121] wait.

[0122] "Aryl" refers to an all-carbon monocyclic or polycyclic ring (including fused rings, spiro rings, or bridged rings) with a conjugated π-electron system, such as, but not limited to, phenyl, naphthyl, phenanthryl, anthraceneyl, fluorenyl, and indeneyl. The aromatic ring can be fused to other cyclic groups (including saturated and unsaturated rings), but cannot contain heteroatoms such as O, N, or S. Furthermore, the point of attachment to the parent group must be on a carbon atom of a ring with a conjugated π-electron system, such as, but not limited to, [other types of rings]. wait.

[0123] "Heteroaryl" refers to an aryl group in which at least one carbon atom on the ring of a conjugated π-electron system is replaced by a heteroatom, which is O, N, or S, such as including but not limited to thienyl, furanyl, isothiazolyl, etc.

[0124] The pharmaceutically acceptable salts described in this invention include acetates, adipates, aspartates, benzoates, benzenesulfonates, bicarbonates, carbonates, bisulfates, sulfates, borates, camphor sulfonates, citrates, cyclohexanesulfonates, ethanedisulfonates, ethanesulfonates, formates, fumarates, glucohepanoates, glucuronates, glucuronates, hexafluorophosphates, hydrochlorides, hydrobromide, hydroiodates, hydroxyethyl sulfonates, lactates, malates, maleic acid esters, malonates, methyl sulfates, naphthates, theosulfonates, nicotinates, nitrates, orotates, oxalates, palmitates, dihydroxyacetate, phosphates, hydrogen phosphates, dihydrogen phosphates, pyroglutamates, glycosides, stearates, succinates, tannins, tartrates, toluenesulfonates, trifluoroacetates, sine sulfonates, methanesulfonates, p-toluenesulfonates, quaternary ammonium salts, or succinates, etc.

[0125] In this invention, the heteroatoms of the heteroaryl and heterocycloalkyl groups are N, O, or S; the number of heteroatoms is 1 to 5.

[0126] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0127] This invention provides a class of pyridone substitutes or their derivatives that exhibit good inhibitory effects on BRD4-BD1 protein. They can be used as novel BET inhibitors for the prevention and / or treatment of various BET-related diseases, such as cancer, with excellent efficacy and promising application prospects.

[0128] Obviously, based on the above description of the present invention, and according to common technical knowledge and conventional methods in the field, various other modifications, substitutions or alterations can be made without departing from the basic technical concept of the present invention.

[0129] The following detailed embodiments further illustrate the above-described content of the present invention. However, this should not be construed as limiting the scope of the present invention to the following examples. All technologies implemented based on the above-described content of the present invention fall within the scope of the present invention. Detailed Implementation

[0130] The raw materials and equipment used in the specific embodiments of the present invention are all known products, obtained by purchasing commercially available products.

[0131] Example 1: Preparation of 6-methyl-4-(6-((4-(methanesulfonyl)piperazin-1-yl)methyl)-4-morpholinothieno[3,2-d]pyrimidin-2-yl)-1,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one (1)

[0132] Synthesis Route 1:

[0133]

[0134] I. Preparation of 4-(2-chlorothiophene[3,2-d]pyrimidin-4-yl)morpholine (IB)

[0135] 2,4-Dichlorothiopheno[3,2-d]pyrimidine (IA, 8.2 g, 40 mmol) was dissolved in methanol (140 mL). Morpholine (7.7 mL, 88 mmol) was slowly added to the solution under stirring at room temperature, and the reaction was stirred at room temperature for 1 hour. The insoluble residue was collected by filtration under reduced pressure. The filter cake was washed successively with water (50 mL × 2) and methanol (30 mL × 1), and then dried in a vacuum drying oven (60 °C) to constant weight to give a white solid product IB (9.55 g, yield 93%). MS-ESI (m / z): 256 [M+H] + .

[0136] II. Preparation of 2-chloro-4-morpholino[3,2-d]pyrimidine-6-carboxaldehyde (IC)

[0137] 4-(2-chlorothiophene[3,2-d]pyrimidin-4-yl)morpholine (IB, 6.4 g, 25 mmol) was dissolved in dry tetrahydrofuran (130 mL) and cooled to -78 °C. Under nitrogen protection, a 1.6 M n-butyllithium solution in n-hexane (19 mL, 30 mmol) was added dropwise. After the addition was complete, the temperature was raised to -60 °C, and the reaction proceeded until a clear brown solution was formed. The reaction solution was then cooled to -78 °C again, and N,N-dimethylformamide (2.9 mL, 37.5 mmol) was slowly added. After the addition was complete, the reaction proceeded at -78 °C for 0.5 hours, and then the temperature was slowly raised to 0 °C (approximately 1–1.5 hours). The reaction solution was slowly poured into 200 mL of 0.25 M dilute hydrochloric acid and 100 mL of ice water. The resulting turbid solution was stirred at 0-10 °C for 0.5 hours. The insoluble matter was collected by vacuum filtration, and the filter cake was washed with 50 mL of ice water. Then, it was dried in a vacuum drying oven (40 °C) to constant weight to obtain a pale yellow solid product IC (6.75 g, yield 95%). MS-ESI (m / z): 284 (M+H) + .

[0138] III. Preparation of 4-(2-chloro-6-(chloromethyl)thieno[3,2-d]pyrimidin-4-yl)morpholine (ID)

[0139] 2-Chloro-4-morpholino[3,2-d]pyrimidin-6-carboxaldehyde (IC, 5.7 g, 20 mmol) was dissolved in methanol (100 mL), cooled to -10 °C, and sodium borohydride (1.14 g, 30 mmol) was added in portions. The reaction was maintained at approximately -10 °C for 1 hour. Then, ice water (90 mL) was slowly added, and the mixture was stirred for 10 minutes. The insoluble matter was collected by vacuum filtration and washed with a small amount of ice water. The filter cake was dried to obtain the intermediate (2-chloro-4-morpholino[3,2-d]pyrimidin-6-yl)methanol. This intermediate was then dissolved in dichloromethane (100 mL), cooled to 0 °C, and thionyl chloride (20 mL) was slowly added with stirring. After the addition was complete, the system was brought to room temperature and reacted for 2 hours. The reaction mixture was evaporated to dryness, then ethanol (30 mL) was added and evaporated again under reduced pressure. This process was repeated twice. Then, ethanol / water ratios of 15 mL / 45 mL were added to the residue, and the mixture was stirred at 10-15°C for 1 hour. The mixture was filtered, and the filter cake was washed with an ice-cold ethanol-water solution (V / V = 1 / 3, 30 mL). The cake was then dried to give a white solid product ID (4.99 g, yield 82%). MS-ESI (m / z): 304 (M+1) + .

[0140] IV. Preparation of 4-(2-chloro-6-((4-(methanesulfonyl)piperazin-1-yl)methyl)thieno[3,2-d]pyrimidin-4-yl)morpholine (IE)

[0141] ID (1.22 g, 4 mmol), 1-methanesulfonylpiperazine (0.98 g, 6 mmol), and potassium carbonate (1.10 g, 8 mmol) were mixed in N,N-dimethylformamide (20 mL) and heated to 80 °C for 2 hours. After the reaction was complete, the solvent was removed by concentration under reduced pressure. The residue was purified by column chromatography to give a white solid intermediate IE (1.69 g, 98% yield). MS-ESI (m / z): 432 [M+H] + .

[0142] V. Preparation of 6-methyl-4-(6-((4-(methanesulfonyl)piperazin-1-yl)methyl)-4-morpholinothieno[3,2-d]pyrimidin-2-yl)-1,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one (1)

[0143]

[0144] IE (47 mg, 0.11 mmol), 6-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxobenzofuran-2-yl)-1,6-dihydro-7H-pyrrolo[2,3-C]pyridin-7-one (47 mg, 0.17 mmol), PdCl2 (DPPF) (8 mg, 0.011 mmol), and sodium carbonate (35 mg, 0.33 mmol) were placed in a 100 mL round-bottom flask and dissolved in 1,4-dioxane / water (8 mL / 2 mL). After purging with nitrogen three times, the reaction was carried out at 95 °C for 3 hours under nitrogen protection. After the reaction was complete, the solvent was removed by concentration under reduced pressure. The residue was purified by column chromatography to give a white solid product 1 (34 mg, yield 57%). 1 H NMR (400MHz, CDCl3): δ9.78(s,1H),8.21(s,1H),7.35–7.32(m,2H),7.31(s,1H),4.07–4.01(m,4H),3.94–3 .90(m,4H),3.89(s,2H),3.77(s,3H),3.35–3.26(m,4H),2.81(s,3H),2.72–2.66(m,4H).HRMS:calculated for C 24 H 29 N7O4S2[(M+H) + ],544.1795;found 544.1793.

[0145] Examples 2-15, Preparation of Compound 2-15

[0146] Following the preparation method of Example 1, compounds 2-15 in Table 1 were synthesized using substituted boronic esters or substituted amines of different types. These substituted boronic esters or amines are commercially available or synthesized using methods known in the literature. The structural formulas and chemical names of compounds 2-15 are shown in Table 1.

[0147] Table 1. Structural identification data of compounds 2-15

[0148]

[0149]

[0150]

[0151]

[0152]

[0153] Example 16: Preparation of 4-(6-((4-(2-hydroxy-2-methylpropionyl)piperazin-1-yl)methyl)-4-morpholinothieno[3,2-d]pyrimidin-2-yl)-6-methyl-1,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one (16)

[0154]

[0155] Compound 7 (61 mg, 0.13 mmol), 2-methyl-2-hydroxypropionic acid (24 mg, 0.23 mmol), HATU (75 mg, 0.2 mmol), and N,N-diisopropylethylamine (84 mg, 0.65 mmol) were dissolved in N,N-dimethylformamide (2 mL) and stirred at room temperature for 1 hour. After the reaction was complete, the mixture was diluted with water (10 mL), extracted twice with ethyl acetate (15 mL), and the organic layers were combined. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, filtered to remove the sodium sulfate solid, concentrated under reduced pressure, and the residue was purified by column chromatography to give product 16 (13 mg, 20% yield) as an off-white solid. 1 H NMR (400MHz, CDCl3): δ10.80(s,1H),8.21(s,1H),7.37(t,J=2.5Hz,1H),7.33(d,J=2.5Hz,1H),7.30(s,1H),4.45(s,1H),4.07– 4.02(m,4H),3.94–3.90(m,4H),3.85(s,2H),3.79(s,3H),3.78–3.73(m,4H),2.61–2.55(m,4H),1.49(s,6H).HRMS:calculated for C 27 H 33 N7O4S[(M+H) + ],552.2387;found 552.2379.

[0156] Examples 17-18, Preparation of Compounds 17-18

[0157] Following the preparation method of Example 16, compounds 17-18 in Table 2 were synthesized using different substituted carboxylic acids. These substituted carboxylic acids were commercially available or synthesized using methods known in the literature. The structural formulas and chemical names of compounds 17-18 are shown in Table 2.

[0158] Table 2. Structural identification data of compounds 17-18

[0159]

[0160]

[0161] Example 19: Preparation of 6-methyl-4-(4-morpholinothieno[3,2-d]pyrimidin-2-yl)-1,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one (19)

[0162]

[0163] IB (52 mg, 0.2 mmol), 6-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxobenzofuran-2-yl)-1,6-dihydro-7H-pyrrolo[2,3-C]pyridin-7-one (61 mg, 0.17 mmol), PdCl2 (DPPF) (8 mg, 0.011 mmol), and sodium carbonate (35 mg, 0.33 mmol) were placed in a 100 mL round-bottom flask and dissolved in 1,4-dioxane / water (8 mL / 2 mL). After purging with nitrogen three times, the reaction was carried out at 95 °C for 3 hours under nitrogen protection. After the reaction was complete, the solvent was removed by concentration under reduced pressure. The residue was purified by column chromatography to give a white solid product 19 (57 mg, yield 78%). 1 H NMR (400MHz, DMSO-d6): δ12.07(s,1H),8.33(s,1H),8.22(d,J=5.4Hz,1H),7.50(d,J=5 .4Hz,1H),7.37(s,1H),7.29(s,1H),4.07-3.94(m,4H),3.87-3.76(m,4H),3.67(s,3H).

[0164] Example 20: Preparation of 6-methyl-4-(6-methyl-4-morpholino[3,2-d]pyrimidin-2-yl)-1,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one (20)

[0165]

[0166] Following the synthesis method of Example 19, 2,4-dichlorothieno[3,2-D]pyrimidine was replaced with 2,4-dichlorothieno[3,2-d]pyrimidine to prepare an off-white solid product 20 (31 mg, yield 41%). 1 H NMR (400MHz, DMSO-d6): δ12.06(s,1H),8.30(s,1H),7.36(t,J=2.7Hz,1H),7.2 5–7.20(m,2H),3.99–3.91(m,4H),3.85–3.77(m,4H),3.66(s,3H),2.62(s,3H).

[0167] Example 21: Preparation of 4-(6-(2-hydroxypropyl-2-yl)-4-morpholinothieno[3,2-d]pyrimidin-2-yl)-6-methyl-1,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one (21)

[0168] Synthesis Route 2:

[0169]

[0170] I. Preparation of 2-(2-chloro-4-morpholino[3,2-d]pyrimidin-6-yl)prop-2-ol (II-A)

[0171] IB (512 mg, 2 mmol) was dissolved in dry tetrahydrofuran solution (8 mL), cooled to -78 °C, and under nitrogen protection, 1.6 M n-butyl / n-hexane solution (1.7 mL, 2.6 mmol) was slowly added. After the addition was complete, the temperature was raised to -60 °C and stirred for 1 hour. The reaction solution was then cooled to -78 °C again, and acetone (275 μL, 6 mmol) was added. After the addition was complete, the mixture was kept at -70 °C and stirred for 2 hours. After the reaction was complete, 1 M hydrochloric acid solution (2 mL) was slowly added to quench the reaction, followed by dilution with water (30 mL). The mixture was extracted twice with ethyl acetate (30 mL), and the extracts were combined and concentrated under reduced pressure. The residue was purified by column chromatography to give a white solid product II-A (514 mg, yield 82%). MS-ESI (m / z): 314 (M+H) + .

[0172] II. Preparation of 4-(6-(2-hydroxypropyl-2-yl)-4-morpholinothieno[3,2-d]pyrimidin-2-yl)-6-methyl-1,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one (21)

[0173]

[0174] Following the synthesis method of Example 19, IB was replaced with II-A to prepare off-white solid product 21 (60 mg, yield 70%). 1 H NMR (400MHz, DMSO-d6): δ12.09(s,1H),8.32(s,1H),7.50–7.10(m,3H),6. 07(s,1H),4.10–3.90(m,4H),3.81(s,3H),3.72–3.55(m,4H),1.60(s,6H).

[0175] Example 22: Preparation of 6-methyl-4-(9-methyl-8-((4-(methanesulfonyl)piperazin-1-yl)methyl)-6-morpholinyl-9H-purine-2-yl)-1,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one (22)

[0176] Synthesis Route 3:

[0177]

[0178] I. Preparation of 2-chloro-9-methyl-6-morpholino-9H-purine-8-carboxaldehyde (III-B)

[0179] Weigh 2.53 g (10 mmol) of 4-(2-chloro-9-methyl-9H-purin-6-yl)morpholine III-A and dissolve it in 150 mL of dry tetrahydrofuran solution. Cool to -78 °C and add N,N,N',N'-tetramethylethylenediamine (2.4 mL, 16 mmol) under nitrogen protection. Then add 1.6 M n-butyllithium / n-hexane solution (10 mL, 16 mmol) dropwise. After the addition is complete, maintain the temperature and stir for 0.5 hours. Then raise the temperature to -40 °C and stir for 1 hour. Cool back to -78 °C and add N,N-dimethylformamide (1.5 mL, 20 mmol), stirring for 2 hours. After the reaction was complete, the reaction was quenched with water (10 mL), the temperature was raised to room temperature, diluted with water (100 mL), and extracted twice with dichloromethane (50 mL). The extracts were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a white solid crude product III-B (1.44 g, yield 51%). MS-ESI (m / z): 282 (M+H) + .

[0180] II. Preparation of 4-(2-chloro-9-methyl-8-(4-(methanesulfonyl)piperazin-1-yl)methyl)-9H-purine-6-yl)morpholine (III-C)

[0181] III-B (564 mg, 2 mmol) was dissolved in 1,2-dichloroethane (10 mL), and 1-methanesulfonylpiperazine (361 mg, 2.2 mmol) was added. One drop of acetic acid was added, and the mixture was stirred at room temperature for 1 hour. Then, sodium triacetoxyborohydride (848 mg, 4 mmol) was added, and the mixture was stirred overnight at room temperature. After the reaction was complete, the reaction solution was diluted with water (20 mL), and extracted twice with dichloromethane (20 mL). The organic layers were combined, concentrated to dryness under reduced pressure, and the residue was purified by column chromatography to give a white solid product III-C (327 mg, 38% yield). MS-ESI (m / z): 430 (M+H) + .

[0182] III. Preparation of 6-methyl-4-(9-methyl-8-((4-(methanesulfonyl)piperazin-1-yl)methyl)-6-morpholinyl-9H-purine-2-yl)-1,6-dihydro-7H-pyrrolo[2,3-c]pyridin-7-one (22)

[0183]

[0184] Following the synthesis method of Example 19, IB was replaced with III-C to prepare off-white solid product 22 (56 mg, yield 52%). 1 H NMR (400MHz, DMSO-d6): δ11.93(s,1H),7.62(s,1H),7.07–6.93(m,2H),4.32(s,2H),4.19– 4.02(m,4H),3.87(s,3H),3.79–3.69(m,4H),3.63(s,3H),2.87(s,3H),2.63–2.54(m,8H).

[0185] The following specific experimental examples demonstrate the beneficial effects of the present invention.

[0186] Experimental Example 1: Affinity test with BRD4-BD1 protein

[0187] The binding of compounds to BRD4-BD1 protein was detected using homogeneous time-resolved fluorescence spectroscopy. (+)-JQ1 was used as a positive control. The BRD4-BD1 time-resolved fluorescence resonance energy transfer (TR-FRET) assay kit (purchased from BPSBioscience) and a multi-functional microplate reader were used.

[0188]

[0189] Experimental procedure:

[0190] (1) Compound preparation: The compound was serially diluted with DMSO, starting at a concentration of 1 μM and then diluted 3 times to obtain 6 concentrations, with duplicate wells for each concentration.

[0191] (2) Prepare reaction solution: Dilute BRD4-BD1 protein and Biotin-labeled histone H4 peptide with Diluent Buffer from the kit.

[0192] (3) Preparation of detection solution: Dilute Anti-GST-TB with the Detection Buffer provided in the kit. 2+ Cryptate and SA-XL-665.

[0193] (4) Take a 384-well plate and arrange it according to the plate layout: test compound well, control well min (high concentration positive drug), control well max (DMSO).

[0194] (5) Add 20 nL of the corresponding concentration of compound or DMSO solution to each well of the plate, then add 10 μL of reaction solution to each well, followed by 10 μL of detection solution.

[0195] (6) After gently shaking and mixing, incubate at room temperature for 60 minutes, and then read the fluorescence signal on the Envision multi-functional microplate reader.

[0196] Data Analysis:

[0197] Calculation formula

[0198]

[0199] Fitting dose-response curve

[0200] Plotting the concentration logarithm on the X-axis and the percentage inhibition rate on the Y-axis, a dose-response curve was fitted using the log(inhibitor) vs. response-variable slope method in GraphPad Prism 5 to derive the IC50 of the compound's inhibition of protein binding. 50 value.

[0201] Table 3 below shows the affinity test activity of the compounds of the present invention for BRD4-BD1 protein. A represents IC50. 50 <100nM, B represents 100nM <IC 50 <1000nM, C represents IC 50 >1000nM.

[0202] Table 3. Affinity test of the compounds of the present invention against BRD4-BD1 protein.

[0203] 1 A 12 A 2 A 13 A 3 C 14 A 4 C 15 A 5 C 16 A 6 A 17 A 7 A 18 A 8 A 19 B 9 A 20 B 10 A 21 A 11 A 22 A

[0204] Table 3 shows that the compounds of the present invention have good inhibitory effects on BRD4-BD1 protein, with compounds 1-2, 6-18, and 21-22 showing better effects.

[0205] Experimental Example 2: Biological assay of the inhibitory effect on the proliferation of lymphoma cells SU-DHL-4 and SU-DHL-6

[0206] ① Dissolve the compound in 100% DMSO to prepare compound stock solutions with concentrations of 1 mM and 0.1 mM.

[0207] ② Add 100 μL of RPMI 1640 complete culture medium to each well of a 96-well plate, add 0.2 μL of 1 mM or 0.1 mM stock solution, and add 0.2 μL of DMSO to the control group.

[0208] ③ After counting SU-DHL-4 and SU-DHL-6 cells, dilute them to 60,000 cells / mL with RPMI 1640 medium and prepare a cell suspension.

[0209] ④ Add 100 μL of diluted cell suspension to each well of a 96-well plate containing the compound.

[0210] ⑤ Place the culture plate in a 37℃, 5% CO2 incubator and incubate for 72 hours.

[0211] ⑥ Add 100 μL of CellTiter-Lumi to each well of a 96-well plate. TM (Beyotime Biotechnology) Luminescence Detection Reagent.

[0212] ⑦ Shake at room temperature for 2 minutes to promote cell lysis.

[0213] ⑧ Let it stand at room temperature away from light for 10 minutes.

[0214] ⑨ Use a multi-functional microplate reader (BioTek Cytation5) to detect chemiluminescence (RLU).

[0215] ⑩ According to the formula

[0216] Table 4 below shows the inhibitory activity of the compounds of the present invention on the proliferation of lymphoma cells SU-DHL-4 and SU-DHL-6.

[0217] Table 4. Inhibitory activity of the compounds of the present invention against the proliferation of lymphoma cells SU-DHL-4 and SU-DHL-6.

[0218]

[0219] Table 4 shows that most of the compounds in this invention have good inhibitory effects on lymphoma cells, and some of the compounds have better inhibitory effects on lymphoma cells than the positive control compound JQ-1.

[0220] In summary, this invention provides a class of pyridone substitutes or their derivatives that exhibit good inhibitory effects on BRD4-BD1 protein. These can serve as novel BET inhibitors for the prevention and / or treatment of various BET-related diseases, such as cancer, with excellent efficacy and promising application prospects.

Claims

1. A compound, its salt, and its stereoisomers, characterized in that: The compound is shown in Formula II: Formula II in, R1 is selected from hydrogen, hydroxyl group, C1~C3 alkyl group, and NR5R6; R5 and R6 connect to the N atom to form substituted or unsubstituted bonds. , , , ; n is 0 or 1; Each R a R b Each is independently selected from hydrogen and C1~C3 alkyl groups; R2 is selected from hydrogen and C1~C3 alkyl groups; X1 is selected from N or CH; X is selected from S or N; Y is selected from CR8 or NR8; The two dashed lines connecting X and Y are either empty or a key depending on the choice between X and Y. When the dashed line is empty, it is a single key; when the dashed line is a key, it is a double key. And the two dashed lines cannot be empty or a key at the same time. R8 is selected from hydrogen and C1-C3 alkyl groups; The , , , Each substituent is independently selected from substituted or unsubstituted C1~C3 alkyl groups, -S(O)(O)R 10 -C(O)R 10 3- to 6-membered cycloalkyl groups and hydroxyl groups; R 10 Selected from substituted or unsubstituted C1-C3 alkyl groups; Each substituent of the alkyl group is independently selected from halogen, hydroxyl, amino, nitro, carboxyl, cyano, and 3- to 6-membered cycloalkyl groups.

2. The compound, its salt, and its stereoisomers according to claim 1, characterized in that: Selected from , ; R8 is selected from hydrogen and C1~C3 alkyl groups.

3. The compound, its salt, and its stereoisomers according to claim 1, characterized in that: The compound is shown in Formula III: Formula III in, R5 and R6 connect to the N atom to form substituted or unsubstituted bonds. , , , ; R2 is selected from C1~C3 alkyl groups; X1 is selected from N or CH; X is selected from S or N; Y is selected from CR8 or NR8; The two dashed lines connecting X and Y are either empty or a key depending on the choice between X and Y. When the dashed line is empty, it is a single key; when the dashed line is a key, it is a double key. And the two dashed lines cannot be empty or a key at the same time. R8 is selected from hydrogen and C1-C3 alkyl groups; The , , , Each substituent is independently selected from substituted or unsubstituted C1~C3 alkyl groups, -S(O)(O)R 10 -C(O)R 10 3- to 6-membered cycloalkyl groups and hydroxyl groups; R 10 Selected from substituted or unsubstituted C1-C3 alkyl groups; Each substituent of the alkyl group is independently selected from halogen, hydroxyl, amino, nitro, carboxyl, cyano, and 3- to 6-membered cycloalkyl groups.

4. The compound, its salt, and its stereoisomers according to claim 3, characterized in that: Selected from , ; R8 is selected from hydrogen and C1~C3 alkyl groups.

5. The compound, its salt, and its stereoisomers according to claim 3, characterized in that: The compound is shown in Formula IV: Formula IV in, X2 is selected from NR', CHR', and O; R' is selected from hydrogen, substituted or unsubstituted C1~C3 alkyl groups, and -S(O)(O)R. 10 -C(O)R 10 3- to 6-membered cycloalkyl groups and hydroxyl groups; R 10 Selected from substituted or unsubstituted C1-C3 alkyl groups; R2 is selected from C1~C3 alkyl groups; X1 is selected from N or CH; X is selected from S or N; Y is selected from CR8 or NR8; The two dashed lines connecting X and Y are either empty or a key depending on the choice between X and Y. When the dashed line is empty, it is a single key; when the dashed line is a key, it is a double key. And the two dashed lines cannot be empty or a key at the same time. R8 is selected from hydrogen and C1-C3 alkyl groups; Each substituent of the alkyl group is independently selected from halogen, hydroxyl, amino, nitro, carboxyl, cyano, and 3- to 6-membered cycloalkyl groups.

6. The compound, its salt, and its stereoisomers according to claim 5, characterized in that: R' is selected from hydrogen, -S(O)(O)R 10 C1~C3 alkyl groups , , , , , 3- to 6-membered cycloalkyl groups and hydroxyl groups; R 10 Selected from C1~C3 alkyl groups; Selected from , ; R8 is selected from hydrogen and C1~C3 alkyl groups.

7. The compound, its salt, and its stereoisomers according to claim 1, characterized in that: The compound is shown in formula V: Formula V in, R1 is selected from hydroxyl groups; R2 is selected from hydrogen and C1~C3 alkyl groups; X1 is selected from N or CH; X is selected from S or N; Y is selected from CR8 or NR8; The two dashed lines connecting X and Y are either empty or a key depending on the choice between X and Y. When the dashed line is empty, it is a single key; when the dashed line is a key, it is a double key. And the two dashed lines cannot be empty or a key at the same time. R8 is selected from hydrogen and C1~C3 alkyl groups.

8. The compound, its salt, and its stereoisomers according to claim 7, characterized in that: Selected from , ; R8 is selected from hydrogen and C1~C3 alkyl groups.

9. A compound, its salt, and its stereoisomers, characterized in that: The compound is one of the following compounds: 。 10. Use of the compound, its salt, or its stereoisomer according to any one of claims 1 to 9 in the preparation of a BET inhibitor and / or in the preparation of a medicament for the prevention and / or treatment of BET-related diseases; wherein the BET inhibitor is a BRD4 inhibitor.

11. The use according to claim 10, characterized in that: The diseases associated with BET are cancer, inflammatory diseases, autoimmune diseases, or viral infections.

12. The use according to claim 11, characterized in that: The BRD4 inhibitor is an inhibitor that inhibits the BRD4-BD1 protein; And / or, the cancer is one or more of the following: lymphoma, leukemia, breast cancer, lung cancer, liver cancer, colon cancer, myeloma, glioma, pancreatic cancer, fibrosarcoma, cervical cancer, and bladder cancer.

13. A pharmaceutical composition, characterized in that: It includes the compounds, salts, and stereoisomers of any one of claims 1 to 9.

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