WO2022033542A1 - 5-membered heteroaromatic pyridone compounds and application thereof - Google Patents

Abstract

A class of 5-membered heteroaromatic pyridone compounds, and specifically disclosed are compounds represented by formula (I) and a pharmaceutically acceptable salt thereof.

Description

5-membered heteroaromatic ring and pyridone compounds and their applications

This application claims the following priority

CN202010817455.8, application date: August 14, 2020;

CN202110532688.8, application date: May 14, 2021.

Field of Invention

The present invention relates to a class of 5-membered heteroaromatic ring and pyridone compounds, in particular to compounds represented by formula (I) and pharmaceutically acceptable salts thereof.

Background technique

BET (bromodomain and extra-terminal domain) is a bromodomain and an additional C-terminal domain. This protein family has 4 members: BRD2, BRD3, BRD4, BRDT, which act as acetylated lysines on histones the "reader". Acetylation of histones is an important way to regulate gene transcription and chromosome structure. BET proteins recognize and bind to acetylated lysines on histones, thereby promoting chromosome decompression and recruiting multiple transcription factors to form a large transcriptional regulatory protein The complex promotes the activation of RNA polymerase II and stimulates the initiation and elongation of transcription, and activates gene transcription, thereby realizing the epigenetic regulation of genes. BET inhibitors can effectively downregulate the gene and protein levels of c-MYC and PD-L1 in tumor cells, thereby realizing the dual functions of tumor proliferation inhibition and immune killing of tumors. Its antitumor activity has been demonstrated.

SUMMARY OF THE INVENTION

The present invention provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereof,

in,

T is selected from N and CH;

T ₁ , T ₂ and T ₃ are each independently selected from N and CH;

R ₁ is selected from -S(=O) ₂ -C _1-3 alkyl, -S(=O)(=NH)-C _1-3 alkyl and -S(=O) ₂ -C _1-3 alkane Amino, the -S(=O) ₂ -C _1-3 alkyl, -S(=O)(=N)-C _1-3 alkyl and -S(=O) ₂ -C _1-3 alkane amino is optionally substituted with 1, 2 or 3 _Ra ;

R ₂ are each independently selected from F, Cl, Br and C _1-3 alkyl optionally substituted with ₁ , 2 or 3 R _b ;

m is selected from 0, 1 and 2;

L ₁ is selected from a single bond, -C(R _c )(R _d )- and -N(R _e )-;

R _a and R _b are each independently selected from F, Cl, Br and I;

_Rc and _Rd are each independently selected from H, F, Cl, Br, I and _CH3 ;

Alternatively, R _c and R _d together with the carbon atoms to which they are attached form a C _3-5 cycloalkyl optionally substituted with 1, 2 or ₃ R;

R _{e is} selected from H and CH ₃ ;

R is selected from F, Cl, Br and I.

In some aspects of the present invention, above, R ₁ is selected from

said

Optionally substituted with 1, 2 or 3 _Ra , other variables are as defined herein.

In some aspects of the present invention, the above R ₁ is selected from

Other variables are as defined in the present invention.

In some embodiments of the present invention, the above R ₂ are independently selected from F, Cl, Br and CH ₃ , and said CH ₃ is optionally substituted with 1, 2 or 3 R _b , and other variables are as defined in the present invention.

In some embodiments of the present invention, the above R ₂ are independently selected from F, and other variables are as defined in the present invention.

In some embodiments of the present invention, the above R _c and R _d together with the carbon atoms to which they are attached constitute a cyclopropyl group optionally substituted with 1, 2 or 3 R, other variables as defined herein.

In some embodiments of the present invention, the above R _c and R _d together with the carbon atoms to which they are attached form a cyclopropyl group, and other variables are as defined in the present invention.

In some aspects of the present invention, the above L ₁ is selected from single bond, -CH ₂ -, -NH- and

Other variables are as defined in the present invention.

In some aspects of the present invention, the above-mentioned structural units R ₁ -L ₁ - are selected from

Other variables are as defined in the present invention.

In some aspects of the present invention, the above-mentioned structural units

selected from

Other variables are as defined in the present invention.

In some schemes of the present invention, the compound represented by the above-mentioned formula (I) or a pharmaceutically acceptable salt thereof,

in,

T is selected from N and CH;

T ₁ , T ₂ and T ₃ are each independently selected from N and CH;

R ₁ is selected from -S(=O) ₂ -C _1-3 alkyl and -S(=O)(=NH)-C _1-3 alkyl, the -S(=O) ₂ -C _{1- 3} alkyl and -S(=O)(=N)-C _1-3 alkyl optionally substituted by 1, 2 or 3 R _a ;

R ₂ are each independently selected from F, Cl, Br and C _1-3 alkyl optionally substituted with ₁ , 2 or 3 R _b ;

m is selected from 0, 1 and 2;

L ₁ is selected from a single bond, -C(R _c )(R _d )- and -N(R _e )-;

R _a and R _b are each independently selected from F, Cl, Br and I;

_Rc and _Rd are each independently selected from H, F, Cl, Br, I and _CH3 ;

Alternatively, _Rc and _Rd together with the atoms to which they are attached form a _{C3-5cycloalkyl} optionally substituted with 1, 2 or ₃ Rs;

R _{e is} selected from H and CH ₃ ;

R is selected from F, Cl, Br and I.

In some aspects of the present invention, the above R ₁ is selected from

said

Optionally substituted with 1, 2 or 3 _Ra , other variables are as defined herein.

In some aspects of the present invention, the above R ₁ is selected from

Other variables are as defined in the present invention.

In some embodiments of the present invention, the above R ₂ are independently selected from F, Cl, Br and CH ₃ , and said CH ₃ is optionally substituted with 1, 2 or 3 R _b , and other variables are as defined in the present invention.

In some embodiments of the present invention, the above R ₂ are independently selected from F, and other variables are as defined in the present invention.

In some embodiments of the present invention, the above R _c and R _d together with the atoms to which they are attached form a cyclopropyl group optionally substituted with 1, 2 or 3 R, other variables as defined herein.

In some embodiments of the present invention, the above R _c and R _d together with the atoms to which they are attached constitute a cyclopropyl group, and other variables are as defined in the present invention.

In some aspects of the present invention, the above L ₁ is selected from single bond, -CH ₂ -, -NH- and

Other variables are as defined in the present invention.

In some aspects of the present invention, the above-mentioned structural units R ₁ -L ₁ - are selected from

Other variables are as defined in the present invention.

In some aspects of the present invention, the above-mentioned structural units

selected from

Other variables are as defined in the present invention.

There are also some solutions of the present invention that are formed by any combination of the above variables.

In some aspects of the present invention, the above-mentioned compound or a pharmaceutically acceptable salt thereof is selected from the group consisting of:

in,

R ₁ and R ₂ are as defined in the present invention.

The present invention also provides a compound represented by the following formula or a pharmaceutically acceptable salt thereof,

technical effect

The compounds of the present invention all have significant BET Bromodomain inhibitory activity; the compounds of the present invention have short half-lives, wider plasma distribution, and moderate bioavailability; the compounds of the present invention exhibit significant tumor-inhibiting effects, and animals tolerated during administration Sex is good.

Definition and Explanation

Unless otherwise specified, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be considered indeterminate or unclear without specific definitions, but should be understood in its ordinary meaning. When a trade name appears herein, it is intended to refer to its corresponding commercial product or its active ingredient.

As used herein, the term "pharmaceutically acceptable" refers to those compounds, materials, compositions and/or dosage forms that, within the scope of sound medical judgment, are suitable for use in contact with human and animal tissue , without excessive toxicity, irritation, allergic reactions or other problems or complications, commensurate with a reasonable benefit/risk ratio.

The term "pharmaceutically acceptable salts" refers to salts of the compounds of the present invention, prepared from compounds with specific substituents discovered by the present invention and relatively non-toxic acids or bases. When compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting such compounds with a sufficient amount of base in neat solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine or magnesium salts or similar salts. When compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting such compounds with a sufficient amount of acid in neat solution or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts including, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, Hydrogen sulfate, hydroiodic acid, phosphorous acid, etc.; and organic acid salts including, for example, acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, Similar acids such as fumaric, lactic, mandelic, phthalic, benzenesulfonic, p-toluenesulfonic, citric, tartaric, and methanesulfonic acids; also include salts of amino acids such as arginine, etc. , and salts of organic acids such as glucuronic acid. Certain specific compounds of the present invention contain both basic and acidic functional groups and thus can be converted into either base or acid addition salts.

The pharmaceutically acceptable salts of the present invention can be synthesized from the acid or base containing parent compound by conventional chemical methods. Generally, such salts are prepared by reacting the free acid or base form of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent or a mixture of the two.

Unless otherwise indicated, the term "isomer" is intended to include geometric isomers, cis-trans isomers, stereoisomers, enantiomers, optical isomers, diastereomers and tautomers isomer.

The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers isomers, (D)-isomers, (L)-isomers, and racemic mixtures thereof and other mixtures, such as enantiomerically or diastereomerically enriched mixtures, all of which belong to this within the scope of the invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl. All such isomers, as well as mixtures thereof, are included within the scope of the present invention.

Unless otherwise indicated, the terms "enantiomers" or "optical isomers" refer to stereoisomers that are mirror images of each other.

Unless otherwise specified, the terms "cis-trans isomer" or "geometric isomer" result from the inability to rotate freely due to double bonds or single bonds to ring carbon atoms.

Unless otherwise indicated, the term "diastereomer" refers to a stereoisomer in which the molecule has two or more chiral centers and the molecules are in a non-mirror-image relationship.

Unless otherwise specified, "(+)" means dextrorotatory, "(-)" means levorotatory, and "(±)" means racemic.

Use solid wedge keys unless otherwise specified

and wedge-dotted keys

Indicate the absolute configuration of a stereocenter, using a straight solid key

and straight dashed keys

Indicate the relative configuration of the stereocenter, with a wavy line

Represents a solid wedge key

or wedge-dotted key

or with wavy lines

Represents a straight solid key

or straight dashed key

Unless otherwise indicated, the terms "enriched in one isomer", "enriched in isomers", "enriched in one enantiomer" or "enriched in one enantiomer" refer to one of the isomers or pairs The enantiomer content is less than 100%, and the isomer or enantiomer content is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or Greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.

Unless otherwise indicated, the terms "isomeric excess" or "enantiomeric excess" refer to the difference between two isomers or relative percentages of two enantiomers. For example, if the content of one isomer or enantiomer is 90% and the content of the other isomer or enantiomer is 10%, the isomer or enantiomeric excess (ee value) is 80% .

Optically active (R)- and (S)-isomers, as well as D and L isomers, can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the present invention is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting mixture of diastereomers is separated and the auxiliary group is cleaved to provide pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a diastereomeric salt is formed with an appropriate optically active acid or base, followed by conventional methods known in the art The diastereoisomers were resolved and the pure enantiomers recovered. In addition, separation of enantiomers and diastereomers is usually accomplished by the use of chromatography employing a chiral stationary phase, optionally in combination with chemical derivatization (eg, from amines to amino groups) formate).

The compounds of the present invention may contain unnatural proportions of atomic isotopes at one or more of the atoms that constitute the compound. For example, compounds can be labeled with radioisotopes, such as tritium ( ³ H), iodine-125 ( ¹²⁵ I) or C-14 ( ¹⁴ C). For another example, deuterated drugs can be formed by replacing hydrogen with deuterium, and the bonds formed by deuterium and carbon are stronger than those formed by ordinary hydrogen and carbon. Compared with non-deuterated drugs, deuterated drugs can reduce toxic side effects and increase drug stability. , enhance the efficacy, prolong the biological half-life of drugs and other advantages. All transformations of the isotopic composition of the compounds of the present invention, whether radioactive or not, are included within the scope of the present invention.

The terms "optional" or "optionally" mean that the subsequently described event or circumstance may, but need not, occur, and that the description includes instances where said event or circumstance occurs and instances in which it does not. .

The term "substituted" means that any one or more hydrogen atoms on a specified atom are replaced by a substituent, which may include deuterium and hydrogen variants, as long as the valence of the specified atom is normal and the substituted compound is stable. When the substituent is oxygen (ie =O), it means that two hydrogen atoms are substituted. Oxygen substitution does not occur on aromatic groups. The term "optionally substituted" means that it may or may not be substituted, and unless otherwise specified, the type and number of substituents may be arbitrary on a chemically achievable basis.

When any variable (eg, R) occurs more than once in the composition or structure of a compound, its definition in each case is independent. Thus, for example, if a group is substituted with 0-2 Rs, the group may optionally be substituted with up to two Rs, with independent options for R in each case. Furthermore, combinations of substituents and/or variants thereof are permissible only if such combinations result in stable compounds.

When the number of a linking group is 0, such as -(CRR) ₀ -, it means that the linking group is a single bond.

When the number of a substituent is 0, it means that the substituent does not exist, such as -A-(R) ₀ means that the structure is actually -A.

When a substituent is vacant, it means that the substituent does not exist. For example, when X in A-X is vacant, it means that the structure is actually A.

When one of the variables is selected from a single bond, it means that the two groups connected to it are directly connected, for example, when L in A-L-Z represents a single bond, it means that the structure is actually A-Z.

When the bond of a substituent can be cross-linked to two or more atoms on a ring, the substituent can bond to any atom on the ring, for example, a structural unit

It means that the substituent R can be substituted at any position on cyclohexyl or cyclohexadiene. When the listed substituents do not indicate through which atom it is attached to the substituted group, such substituents may be bonded through any of its atoms, for example, pyridyl as a substituent may be through any one of the pyridine rings. The carbon atom is attached to the substituted group.

When the listed linking group does not indicate its direction of attachment, the direction of attachment is arbitrary, for example,

The linking group L in the middle is -MW-, at this time -MW- can connect ring A and ring B in the same direction as the reading order from left to right.

It is also possible to connect ring A and ring B in the opposite direction to the reading order from left to right.

Combinations of the linking groups, substituents and/or variants thereof are permissible only if such combinations result in stable compounds.

Unless otherwise specified, when a group has one or more attachable sites, any one or more sites in the group can be linked to other groups by chemical bonds. When the connection method of the chemical bond is not located, and there is an H atom at the linkable site, when the chemical bond is connected, the number of H atoms at the site will be correspondingly reduced with the number of chemical bonds connected to the corresponding valence. the group. The chemical bond connecting the site to other groups can be represented by straight solid line bonds

straight dotted key

or wavy lines

Express. For example, a straight solid bond in -OCH ₃ indicates that it is connected to other groups through the oxygen atom in this group;

The straight dashed bond in the group indicates that it is connected to other groups through the two ends of the nitrogen atom in the group;

The wavy line in the phenyl group indicates that it is connected to other groups through the 1 and 2 carbon atoms in the phenyl group;

Indicates that any linkable site on the piperidinyl group can be connected to other groups through a chemical bond, including at least

These 4 connection methods, even if the H atom is drawn on -N-, but

still includes

For the group in this connection mode, when one chemical bond is connected, the H at the site will be correspondingly reduced by one to become the corresponding monovalent piperidinyl group.

Unless otherwise specified, the number of atoms in a ring is generally defined as the number of ring members, eg, "5-7 membered ring" refers to a "ring" of 5-7 atoms arranged around it.

Unless otherwise specified, the term "C _1-3 alkyl" is used to denote a straight or branched chain saturated hydrocarbon group consisting of 1 to 3 carbon atoms. The C _1-3 alkyl group includes C _1-2 and C _2-3 alkyl groups, etc.; it can be monovalent (eg methyl), divalent (eg methylene) or multivalent (eg methine) . Examples of _C1-3 alkyl groups include, but are not limited to, _methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), and the like.

Unless otherwise specified, the term "C _1-3 alkylamino" refers to those alkyl groups containing 1 to 3 carbon atoms attached to the remainder of the molecule through an amino group. The C _1-3 alkylamino groups include C _1-2 , C ₃ and C ₂ alkylamino groups and the like. Examples of C _1-3 alkylamino include, but are not limited to, -NHCH ₃ , -N(CH ₃ ) ₂ , -NHCH ₂ CH ₃ , -N(CH ₃ )CH ₂ CH ₃ , -NHCH ₂ CH ₂ CH ₃ , - NHCH ₂ (CH ₃ ) ₂ and the like.

Unless otherwise specified, "C _3-5 cycloalkyl" means a saturated cyclic hydrocarbon group consisting of 3 to 5 carbon atoms, which is a monocyclic ring system, said C _3-5 cycloalkyl including C _{3 -4} and C _4-5 cycloalkyl, etc.; it may be monovalent, divalent or polyvalent. Examples of _C3-5 cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and the like.

The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments enumerated below, embodiments formed in combination with other chemical synthesis methods, and those well known to those skilled in the art Equivalent to alternatives, preferred embodiments include, but are not limited to, the embodiments of the present invention.

The structure of the compound of the present invention can be confirmed by conventional methods well known to those skilled in the art. If the present invention relates to the absolute configuration of the compound, the absolute configuration can be confirmed by conventional technical means in the art. For example, single crystal X-ray diffraction method (SXRD), the cultured single crystal is collected by Bruker D8 venture diffractometer, the light source is CuKα radiation, and the scanning mode is:

After scanning and collecting relevant data, the crystal structure was further analyzed by the direct method (Shelxs97), and the absolute configuration could be confirmed.

The solvent used in the present invention is commercially available.

The present invention adopts the following abbreviations: aq represents water; eq represents equivalent, equivalent; DCM represents dichloromethane; PE represents petroleum ether; DMF represents N,N-dimethylformamide; DMSO represents dimethyl sulfoxide; EtOAc represents ethyl acetate; EtOH represents ethanol; MeOH represents methanol; Cbz represents benzyloxycarbonyl, which is an amine protecting group; BOC represents tert-butoxycarbonyl, which is an amine protecting group; HOAc represents acetic acid; rt represents room temperature; O/N for overnight; THF for tetrahydrofuran; Boc ₂ O for di-tert-butyl dicarbonate; TFA for trifluoroacetic acid; DIPEA for diisopropylethylamine; TEA for triethylamine; iPrOH for 2-propanol ; mp represents melting point; AcOH represents acetic acid; Pd(dppf)Cl ₂ represents [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride; Pd(OAc) ₂ represents palladium acetate; DBU represents 1,8-Diazabicycloundec-7-ene.

Compounds are named according to conventional nomenclature in the art or are used

Software naming, commercially available compounds use supplier catalog names.

detailed description

The present invention will be described in detail by the following examples, but it does not mean any unfavorable limitation of the present invention. The present invention has been described in detail herein, and specific embodiments thereof have also been disclosed. For those skilled in the art, various changes and modifications can be made to the specific embodiments of the present invention without departing from the spirit and scope of the invention. will be obvious.

Example 1

synthetic route:

Step 1: Synthesis of Compounds 1-2

Compound 1-1 (5g, 45.01mmol, 1eq) was added to a mixture of ethylene glycol monomethyl ether (100mL) and hydrazine hydrate (10.30g, 174.89mmol, 10mL, purity 85%, 3.89eq) under nitrogen protection Stir at 110°C for 20 hours. The reaction solution was cooled to room temperature and solid was precipitated, filtered, and the filter cake was dried to obtain compound 1-2. ¹ H NMR (400MHz, CDCl ₃ )δ10.28(br s,1H),7.52(s,1H),6.95(d,J=7.2Hz,1H),5.63(d,J=6.8Hz,1H), 5.38 (d, J=2.0 Hz, 1H), 4.09 (s, 2H).

Step 2: Synthesis of Compounds 1-3

Compound 1-2 (3.5 g, 27.97 mmol, 1 eq) and propionaldehyde (2.79 g, 48.09 mmol, 3.5 mL, 1.72 eq) were added to ethanol (100 mL) and stirred at 80° C. for 4 hours. The reaction solution was cooled to 0°C, a solid was precipitated, filtered, and the residual solvent was removed from the filter cake under reduced pressure to obtain compound 1-3. ¹ H NMR (400MHz, DMSO-d ₆ )δ10.55(br s,1H),10.11(br s,1H),7.28(t,J=4.8Hz,1H),7.10(d,J=7.2Hz, 1H), 5.84 (d, J=3.6Hz, 1H), 5.55 (s, 1H), 2.20-2.27 (m, 2H), 1.04 (t, J=7.2Hz, 3H).

Step 3: Synthesis of Compounds 1-4

Compound 1-3 (1.0 g, 6.05 mmol, 1 eq) was dissolved in diphenyl ether (30 mL) and stirred at 220° C. for 6 hours. The reaction solution was cooled to room temperature, and a solid was precipitated, which was filtered. The filter cake was dissolved in methanol (20 mL) and filtered to remove a small amount of black insoluble matter. The filtrate was spin-dried, the obtained crude product was slurried with methanol (5 mL) at room temperature for 10 minutes, and filtered to obtain compound 1-4. LCMS (ESI) m/z: 148.8 [M+1] ⁺ ; ¹ H NMR (400 MHz, DMSO-d ₆ ) δ 11.00 (br s, 1H), 10.51 (br s, 1H), 6.87 (d, J = 2.4 Hz, 1H), 6.76 (s, 1H), 6.27 (d, J = 6.8 Hz, 1H), 2.31 (s, 3H).

Step 4: Synthesis of Compounds 1-6

Compound 1-5 (10g, 39.92mmol, 1eq) was dissolved in DMSO (300mL), and then 2,4-difluorophenol (4.00g, 30.75mmol, 0.77eq) and potassium carbonate (10.00g, 72.36mmol, 2,4-difluorophenol) were added. 1.81eq), and reacted at 100°C for 5 hours after the addition was completed. Water (500 mL) was added to the reaction solution, then extracted with ethyl acetate (200 mL*3), the combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the solution was concentrated under reduced pressure to obtain the compound 1-6. LCMS (ESI) m/z: 343.7 [M+1] ⁺ .

Step 5: Synthesis of Compounds 1-7

Compound 1-6 (8.6g, 24.99mmol, 1eq) was dissolved in ethanol (70mL), cooled to 0°C, sodium borohydride (1.89g, 50.09mmol, 2eq) was added, and the temperature was raised to 80°C for 3 hours after the addition. . The reaction solution was quenched by adding water (100 mL), then extracted with ethyl acetate (100 mL*3), the combined organic phases were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column (petroleum ether/ethyl acetate=3/1) to obtain compound 1-7. LCMS (ESI) m/z: 317.6 [M+1] ⁺ .

Step 6: Synthesis of Compounds 1-8

Phosphorus tribromide (2.02 g, 7.37 mmol, 99% purity, 1 eq) was added dropwise to a solution of compound 1-7 (2.33 g, 7.37 mmol, 1 eq) in dichloromethane (30 mL) at 0°C, and added dropwise. The reaction was completed at 20°C for 3 hours. Water (20 mL) was slowly added dropwise to the constantly stirring reaction solution to quench the reaction, and then extracted with dichloromethane (20 mL*3). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, and filtered. , the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column (petroleum ether/ethyl acetate=10/1) to obtain compound 1-8. LCMS (ESI) m/z: 379.9 [M+1] ⁺ ; ¹ H NMR (400 MHz, CDCl ₃ ) δ 7.93 (S, 1H), 7.92 (S, 1H), 7.12-7.18 (m, 1H), 6.84-6.87 (m, 2H), 4.32 (s, 2H).

Step 7: Synthesis of Compounds 1-9

To the solution of compound 1-8 (2.34g, 6.17mmol, 1eq) dissolved in ethanethiol (2.62g, 42.18mmol, 3.12mL, 6.83eq) was added cesium carbonate (3.04g, 9.34mmol, 1.51eq), added The reaction was completed at 20°C for 16 hours. The reaction was connected to a tail gas absorption device, and all the solvent was pumped into a 4M sodium hydroxide aqueous solution absorption bottle under reduced pressure at room temperature. The obtained crude products 1-9 were directly used in the next reaction without treatment.

Step 8: Synthesis of Compounds 1-10

To a solution of compound 1-9 (2.22g, 6.16mmol, 1eq) dissolved in methanol (30mL), a solution of potassium monopersulfate (11.4g, 18.54mmol, 3.01eq) in water (30mL) was added. The reaction was carried out at 20°C for 1 hour. Potassium monopersulfate (7.6g, 12.36mmol, 2.01eq) was added again and the reaction was continued for 2 hours at 20°C. Water (50 mL) was slowly added to the constantly stirring reaction solution, and a solid was precipitated, which was filtered, and the filter cake was purified by silica gel column (petroleum ether/ethyl acetate=10/1) to obtain compound 1-10. LCMS (ESI) m/z: 393.8 [M+1] ⁺ ; ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.08 (d, J=2.0 Hz, 1 H), 7.98 (d, J=2.0 Hz, 1 H) , 7.21-7.23 (m, 1H), 6.93-6.98 (m, 2H), 4.10 (s, 2H), 2.95 (q, J=7.6Hz, 2H), 1.42 (t, J=7.6Hz, 3H).

Step 9: Synthesis of Compound 1

To a solution of compound 1-10 (50mg, 337.47μmol, 1eq) and compound 1-4 (137mg, 349.30μmol, 1.04eq) dissolved in 1,4-dioxane (3mL) was added potassium phosphate (145mg, 683.11 μmol, 2.02eq), cuprous iodide (7mg, 36.76μmol, 1.09e-1eq) and trans-1,2-cyclohexanediamine (5mg, 43.79μmol, 5.37μL, 0.13eq), the reaction was carried out at 110°C under stirring for 40 hours. The reaction solution was diluted with ethyl acetate (10 mL), washed with water (5 mL) and saturated brine (5 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by chromatography plate (dichloromethane/methanol=10/1) to obtain compound 1. LCMS (ESI) m/z: 459.8 [M+1] ⁺ ; 1H NMR (400 MHz, CDCl ₃ ) δ 9.74 (br s, 1H), 8.10 (s, 1H), 7.93 (s, 1H), 7.15- 7.21(m, 1H), 6.92-7.07(m, 4H), 6.39(d, J=8.0Hz, 1H), 4.23(s, 2H), 3.06(q, J=8.0Hz, 2H), 2.57(s , 3H), 1.48 (t, J=8.0Hz, 3H).

Example 2

synthetic route:

Step 1: Synthesis of Compound 2-2

Compound 2-1 (2 g, 10.47 mmol, 1 eq) and ethylsulfonyl chloride (1.4 g, 10.89 mmol, 1.03 mL, 1.04 eq) were added to pyridine (10 mL) and stirred at 26° C. for 5 hours. Ethyl acetate (50 mL) and water (50 mL) were added to the reaction solution, the pH of the aqueous phase was adjusted to 6 with 1M hydrochloric acid solution, the organic phase was separated and washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. Compound 2-2 was obtained.

Step 2: Synthesis of Compounds 2-3

To a solution of compound 2-2 (1g, 3.53mmol, 1eq) and 2,4-difluorophenol (460mg, 3.54mmol, 1.00eq) dissolved in N,N-dimethylformamide (10mL) was added cesium carbonate (2.4g, 7.37mmol, 2.09eq), the reaction was reacted under microwave at 150°C for 4 hours. The reaction solution was treated with water (30 mL), then extracted with ethyl acetate (20 mL*3), the combined organic phases were washed with water (20 mL) and saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude product was filtered through silica gel Column purification (petroleum ether/ethyl acetate=3/1) gave compound 2-3. LCMS (ESI) m/z: 392.9, 394.9 [M+1] ⁺ .

Step 3: Synthesis of Compound 2

Compound 1-4 (65mg, 438.71μmol, 1eq) and compound 2-3 (180mg, 457.78μmol, 1.04eq) were dissolved in dioxane (3mL), followed by potassium phosphate (200.00mg, 918.66μmol, 2.09 eq), (1R,2R)-1,2-cyclohexanediamine (12mg, 105.08μmol, 0.24eq), cuprous iodide (15mg, 78.8μmol, 0.18eq), and the temperature was raised to 110° C. to react for 32 hours. The reaction solution was diluted with ethyl acetate (10 mL) and filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by preparative chromatography (chromatographic column: Phenomenex Gemini-NX 80*30mm*3 μm; mobile phase: [water (10mM NH ₄ HCO ₃ ) -ACN]; acetonitrile %: 43%-73%, 9 min) to give compound 2. LCMS (ESI) m/z: 461.0 [M+1] ⁺ ; ¹ H NMR (400 MHz, CDCl ₃ ) δ 9.07 (br s, 1H), 8.21 (d, J=2.5 Hz, 1H), 7.78 (dd , J＝4.0, 8.0Hz, 1H), 7.2(s, 1H), 7.14-7.19(m, 1H), 7.0(d, J＝8.0Hz, 1H), 6.98-6.91(m, 2H), 6.83( s, 1H), 3.35 (q, J=7.4 Hz, 2H), 2.49 (s, 3H), 1.51 (t, J=8.0 Hz, 3H).

Example 3

synthetic route:

Step 1: Synthesis of Compound 3-1

To a solution of compound 1-8 (2.4g, 6.33mmol, 1eq) dissolved in ethanol (10mL) and water (5mL) was added sodium sulfite (860mg, 6.82mmol, 1.08eq), and the reaction was stirred at 80°C for 15 hours. The solvent was removed from the reaction solution under reduced pressure, the residue was diluted with water (20 mL) and washed with dichloromethane (20 mL*2), and the aqueous phase was freeze-dried to obtain compound 3-1, which was directly used in the next reaction.

Step 2: Synthesis of Compound 3-2

To a solution of compound 3-1 (2.5g, 6.58mmol, 1eq) in dichloromethane (5mL) at 0°C was added N,N-dimethylformamide (95.00mg, 1.30mmol, 0.1mL, 1.98e-1eq) and oxalyl chloride (1.45 g, 11.42 mmol, 1 mL, 1.74 eq), and the reaction was stirred at 24 °C for 13 hours. The reaction solution was directly spin-dried to obtain compound 3-2, which was directly used in the next reaction. LCMS (ESI) m/z: 397.4 [M+1] ⁺ .

Step 3: Synthesis of Compound 3-3

To a solution of compound 3-2 (200mg, 501.75μmol, 1eq) and N,N-dimethylamine (150mg, 1.84mmol, 168.54μL, 3.67eq, HCl) in dichloromethane (5mL) at 0°C was added 4-N , N-dimethylaminopyridine (17 mg, 139.15 μmol, 2.77e-1 eq) and DBU (808.00 mg, 5.31 mmol, 800.00 μL, 10.58 eq), and the reaction was stirred at 24° C. for 3 hours. The reaction solution was diluted with dichloromethane (10 mL), and adjusted to pH=6 with 1M HCl, the organic phase was separated, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude product was purified with a preparative plate (dichloromethane/methanol=10/1) to obtain compound 3-3. LCMS (ESI) m/z: 407.0, 409.0 [M+1] ⁺ ; 1H NMR (400 MHz, CDCl ₃ ) δ 8.09 (d, J=2.0 Hz, 1H), 7.98 (d, J=2.0, 1H) , 7.26-7.22 (m, 1H), 7.01-6.93 (m, 2H), 4.12 (s, 2H), 2.84 (s, 6H).

Step 4: Synthesis of Compound 3

To the solution of compound 3-3 (160mg, 392.90μmol, 1eq) and compound 1-4 (80mg, 539.95μmol, 1.37eq) dissolved in dioxane (10mL) was added potassium phosphate (210mg, 989.31μmol, 2.52eq) ), cuprous iodide (16mg, 84.01μmol, 2.14e-1eq) and (1R,2R)-1,2-cyclohexanediamine (16mg, 140.12μmol, 17.19μL, 3.57e-1eq), the reaction was carried out at 110 Stir at °C for 35 hours. The reaction solution was directly filtered, the filter cake was washed with methanol (10 mL), and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative plate (dichloromethane/methanol=10/1), and the crude product was further purified by preparative chromatography (chromatographic column: Welch Xtimate C18 150*25mm*5μm; mobile phase: [water (10mM NH ₄ HCO ₃ ) -ACN]; acetonitrile %: 33%-63%, 9 min) to give compound 3. LCMS (ESI) m/z: 475.2 [M+1] ⁺ ; ¹ H NMR (400 MHz, CDCl ₃ ) δ 11.07 (br s, 1H), 8.01 (d, J=2.0 Hz, 1H), 7.85 (d , J＝2.0, 1H), 7.15-7.05(m, 2H), 6.92(s, 1H), 6.89-6.82(m, 2H), 6.41(d, J＝8.0Hz, 1H), 4.10(s, 2H) ), 2.82(s, 6H), 2.48(s, 3H).

Example 4

synthetic route:

Step 1: Synthesis of Compound 4-1

To compound 3-2 (2.2g, 5.52mmol, 1eq) was added methylamine (10g, 96.60mmol, 30% purity, 17.50eq) (~30% ethanol solution) at 24°C, and the reaction was stirred for 3 hours. The reaction solution was directly concentrated, the residue was dissolved in ethyl acetate (20 mL) and washed with water (10 mL) and saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under pressure, and the crude product was purified by silica gel column (dichloromethane). Methane/methanol=10/1) to obtain compound 4-1. LCMS (ESI) m/z: 392.9, 394.9 [M+1] ⁺ .

Step 2: Synthesis of Compound 4

To the dioxane of cuprous iodide (38.33mg, 201.28μmol, 3.44e-1eq), compound 4-1 (230mg, 584.94μmol, 1eq) and compound 1-4 (95.83mg, 646.82μmol, 1.11eq) (10mL) potassium phosphate (316.25mg, 1.49mmol, 2.55eq) and (1R,2R)-1,2-cyclohexanediamine (19.17mg, 167.85μmol, 20.59μL, 2.87e-1eq) were added to the mixture, The reaction was stirred under nitrogen at 110°C for 35 hours. The reaction solution was directly filtered, the filtrate was concentrated under reduced pressure, the obtained residue was purified by silica gel column (dichloromethane/methanol=10/1), and the obtained crude product was further purified by preparative chromatography (chromatographic column: Phenomenex Gemini-NX 80*40mm *3 μm; mobile phase: [water (0.05% NH ₃ H ₂ O)-ACN]; % acetonitrile: 26%-56%, 8 min) to give compound 4. LCMS (ESI) m/z: 461.2 [M+1] ⁺ ; ¹ H NMR (400 MHz, CDCl ₃ ) δ 8.12 (s, 1H), 7.95 (s, 1H), 7.26-7.24 (m, 1H), 7.12(d,J＝8.0Hz,1H),6.94-6.89(m,2H),6.77(s,1H),6.47(d,J＝8.0Hz,1H),4.52-4.55(m,1H),4.25 (s, 2H), 2.82 (s, 3H), 2.50 (s, 3H).

biological test

Experimental Example 1. Detection of BRD4 biochemical activity of the compounds of the present invention

1. Experiment Preparation

Test 1: _IC50 characterization of compounds at 10 concentrations.

Compounds were evaluated for _IC50 at 10 concentrations against two BRDs (BRD4-1, BRD4-2), single well, starting concentration of 10 [mu]M, 3-fold serial dilution.

2. Test conditions

BRD buffer composition: 50 mM HEPES-HCl, pH 7.5, 100 mM NaCl, 0.1% BSA, 0.05% CHAPS and 1% DMSO.

Ligand: Histone H4 peptide (1-21) K5/8/12/16Ac-Biotin

Detection: AlphaScreen binding experiment (Ex/Em=680/520-620nm)

experiment procedure:

2.1 Add 4X BRD to the wells of the reaction plate, except for the control wells without BRD, replace with buffer.

2.2 Compounds in 100% DMSO were added to the BRD mixture using Acoustic Technology (Echo550, nanoliter). Centrifuge, shake gently at room temperature, and pre-incubate for 30 minutes.

2.3 Add 4X ligand, centrifuge and shake.

2.4 Gently shake and incubate at room temperature for 30 minutes.

2.5 Add 4X donor beads (donor beads) in the dark. Centrifuge and shake.

2.6 Add 4X acceptor beads (acceptor beads) in the dark. Centrifuge and shake. Protect from light and shake for 60 minutes.

2.7 Alpha detection using Enspire (Ex/Em=680/520-620nm)

3. Experimental results

Table 1 BRD4 detection IC ₅₀ test results

compound	BRD4 (BD1, BD2), IC50 (nM)
1	6.21, 1.44

Conclusion: The compounds of the present invention all have significant BET Bromodomain inhibitory activity.

Experimental example 2. Pharmacokinetic study of the compounds of the present invention

1. Summary

1.1 Male CD-1 mice were used as the test animals, and the LC/MS/MS method was used to determine the drug concentrations in the plasma of mice at different times after intravenous and intragastric administration of the test compounds respectively. To study its pharmacokinetic behavior in mice and evaluate its pharmacokinetic characteristics.

2. Experimental protocol

2.1 Test drug: test compound.

2.2 Drug preparation

Weigh an appropriate amount of sample, add a solvent, stir and ultrasonicate until it becomes clear for intravenous administration.

Weigh an appropriate amount of sample, add a solvent, stir and ultrasonicate to a clear state for intragastric administration.

2.3 Administration

Four male CD-1 mice were divided into two groups. After an overnight fast, one group was administered intravenously, and the other group was administered intragastrically.

3. Operation

Male CD-1 mice were intravenously administered with test compounds, and 30 μL of blood was collected at 0.0833, 0.25, 0.5, 1, 2, 4, 8 and 24 hours, respectively, and placed in commercial test tubes containing EDTA-K ₂ . After the test compound was administered to the gavage group, 30 μL of blood was collected at 0.25, 0.5, 1, 2, 4, 6, 8 and 24 hours, respectively, and placed in a commercial test tube containing EDTA-K ₂ . The tubes were centrifuged at 3000g for 15 minutes to separate the plasma and stored at -60°C. Animals were allowed to eat 2 hours after dosing.

LC/MS/MS method was used to determine the content of the tested compounds in the plasma of mice after intravenous and intragastric administration. The linear range of the method was 2.00-6000 nmol/L; plasma samples were analyzed after acetonitrile precipitation.

4. Pharmacokinetic parameters results

Table 2 Summary of pharmacokinetic parameter data

"--":without;

Experimental conclusion: the half-life of the compound of the present invention is short, the distribution outside the plasma is wide, and the bioavailability is moderate.

Experimental Example 3. In vivo efficacy study of the compounds of the present invention in the MC38 mouse colon cancer cell animal xenograft model

1. Experimental Design

Table 3 Animal groupings and dosing schedules for in vivo pharmacodynamic experiments

group

number of animals

Compound therapy

Dosage (mg/kg)

Dosing volume parameters (μL/g)

Route of administration

Dosing frequency

1	7	vehicle group	--	10	p.o.	BID
2	7	Compound 1	20	10	p.o.	BID

Note: p.o: oral; BID: twice a day.

2. Experimental materials

2.1 Experimental animals

Species: mouse.

Strain: C57 mouse.

Age and weight: 7-8 weeks old.

Gender: Female.

Supplier: Shanghai Sipple-Bikai Laboratory Animal Co., Ltd.

3. Experimental methods and steps

3.1 Cell Culture

Name: MC38 (mouse colon cancer cells).

Source: Cells were purchased from Saibaikang (Shanghai) Biotechnology Co., Ltd., and were maintained and passaged by Shanghai WuXi AppTec New Drug Development Co., Ltd.

Passaging conditions: the culture medium was DMEM medium containing 10% fetal bovine serum, and the culture conditions were 37° C., 5% carbon dioxide. 1:5 was passed every other day, 3 to 4 times a week, and it was the 18th generation at the time of inoculation.

3.2 Tumor cell inoculation

Cells were resuspended in DPBS at a density of ² x 106 cells/mL. 0.1 mL of DPBS (containing 2×10 ⁵ MC38 cells) was subcutaneously inoculated into the right back of each mouse. When the average tumor volume reached (72.97±4.6) mm ³ , the mice were randomly divided into groups and administered according to the tumor volume.

3.3 Tumor measurement and experimental indicators

Tumor diameters were measured with vernier calipers twice a week. The calculation formula of tumor volume is: V=0.5×a×b ² , a and b represent the long and short diameters of the tumor, respectively.

The antitumor efficacy of the compounds was evaluated by TGI (%) or relative tumor proliferation rate T/C (%). Relative tumor proliferation rate T/C (%) = T _RTV /C _RTV × 100 (T _RTV : the average RTV of the treatment group; C _RTV : the average RTV of the negative control group). The relative tumor volume (relative tumor volume, RTV) is calculated according to the results of tumor measurement, and the calculation formula is RTV=V _t /V ₀ , where V ₀ is the tumor volume measured during group administration (ie D0), and V _t is a certain The tumor volume at one measurement, T _RTV and C _RTV were taken on the same day.

TGI (%), reflecting tumor growth inhibition rate. TGI(%)=[(1-(average tumor volume at the end of administration of a certain treatment group-average tumor volume at the beginning of administration of this treatment group))/(average tumor volume at the end of treatment in the solvent control group-starting treatment in the solvent control group time average tumor volume)] × 100.

After the experiment, the tumor weight will be detected, and the percentage of T _weight /C _weight will be calculated, and T _weight /C _weight represents the tumor weight of the administration group and the vehicle control group, respectively.

3.4 Statistical analysis

Statistical analysis was performed using SPSS software based on relative tumor volume and tumor weight at the end of the trial. The comparison between two groups was analyzed by t-test, and the comparison between three or more groups was analyzed by one-way ANOVA. P<0.05 considered significant difference.

4. Experimental conclusion

The results are shown in Table 4.

Table 4

group	Dose (mpk)	TGI(%)	P value
Compound 1	20	77.7	0.001

Conclusion: After 15 days of administration observation, compared with the vehicle control group, the compound of the present invention exhibits a significant tumor-inhibiting effect; and during the administration period, the animals are well tolerated.

Claims (12)

1. A compound represented by formula (I) or a pharmaceutically acceptable salt thereof,

   

   in,

   T is selected from N and CH;

   T ₁ , T ₂ and T ₃ are each independently selected from N and CH;

   R ₁ is selected from -S(=O) ₂ -C _1-3 alkyl, -S(=O)(=NH)-C _1-3 alkyl and -S(=O) ₂ -C _1-3 alkane Amino, the -S(=O) ₂ -C _1-3 alkyl, -S(=O)(=N)-C _1-3 alkyl and -S(=O) ₂ -C _1-3 alkane amino is optionally substituted with 1, 2 or 3 _Ra ;

   R ₂ are each independently selected from F, Cl, Br and C _1-3 alkyl optionally substituted with ₁ , 2 or 3 R _b ;

   m is selected from 0, 1 and 2;

   L ₁ is selected from a single bond, -C(R _c )(R _d )- and -N(R _e )-;

   R _a and R _b are each independently selected from F, Cl, Br and I;

   _Rc and _Rd are each independently selected from H, F, Cl, Br, I and _CH3 ;

   Alternatively, R _c and R _d together with the carbon atoms to which they are attached form a C _3-5 cycloalkyl optionally substituted with 1, 2 or ₃ R;

   R _{e is} selected from H and CH ₃ ;

   R is selected from F, Cl, Br and I.
2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R ₁ is selected from

   

   

   said

   

   Optionally substituted with 1, 2 or 3 _Ra .
3. The compound according to claim 2 or a pharmaceutically acceptable salt thereof, wherein R ₁ is selected from

   

   
4. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R ₂ is independently selected from F, Cl, Br and CH ₃ , wherein CH ₃ is optionally substituted with 1, 2 or 3 R _b .
5. The compound of claim 4, or a pharmaceutically acceptable salt thereof, wherein R ₂ is each independently selected from F.
6. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein R _c and R _d together with the carbon atom to which they are attached form a cyclopropyl group optionally surrounded by 1, 2 or 3 R replace.
7. The compound of claim 6, or a pharmaceutically acceptable salt thereof, wherein _Rc and _Rd together with the carbon atom to which they are attached form a cyclopropyl group.
8. The compound according to claim 1 or 7 or a pharmaceutically acceptable salt thereof, wherein L ₁ is selected from a single bond, -CH ₂ -, -NH- and

   
9. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the structural units R ₁ -L ₁ - are selected from

   

   
10. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the structural unit

    

    selected from

    

    
11. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, which compound is selected from the group consisting of:

    

    in,

    R ₁ and R ₂ are as defined in claim 1 .
12. A compound represented by the following formula or a pharmaceutically acceptable salt thereof,