β-Lactam Compounds and Their Preparation Methods

Abstract

The present invention provides a lactam compound represented by formula (II), a preparation method thereof, and its use as an intermediate in the preparation of an anti-tumor compound represented by formula (A).

Description

[0001] This invention claims the priority of a prior application titled "Lactam Compounds and Their Preparation Methods" with the patent application number 202011286215.6, which was filed with the China National Intellectual Property Administration on November 17, 2020. The full text of the above prior application is incorporated into this invention by reference. Technical Field

[0002] This invention belongs to the field of pharmaceutical chemical synthesis, and particularly relates to a lactam compound and its preparation method. Background Art

[0003] Methionine adenosyltransferase (MAT), also known as S-adenosylmethionine synthetase, is a class of enzymes that can catalyze the reaction of methionine (Met) and ATP to generate S-adenosyl-L-methionine (SAM). SAM is the main methyl donor in the body and can regulate gene expression, transcription, and translation through transmethylation reactions, thereby having an important impact on cell growth, death, and differentiation. Moreover, SAM is also involved in the biosynthesis of polyamines and glutathione.

[0004] MAT enzymes mainly have three subtypes, MAT1A, MAT2A, and MAT2B. MAT1A mainly exists in normal liver cells, while MAT2A is widely distributed in extrahepatic cells. These two subtypes differ in catalytic efficiency and regulation mode. MAT2B does not have the ability to catalyze the synthesis of SAM, but rather serves as a regulatory subunit of MAT2A. After forming a complex with MAT2A, it regulates the catalytic activity of MAT2A.

[0005] Research indicates that in liver cancer cells, the expression level of MAT1A is downregulated and the expression of MAT2A increases, thereby promoting the proliferation of liver cancer cells. In addition, the phenomenon of abnormally elevated MAT2A expression levels also exists in many other types of tumors, and silencing the gene encoding MAT2A can lead to the death of cancer cells. Further, Marjon et al. (Cell Reports 15(3)(2016)574–587) found that cancer cell lines lacking MTAP are sensitive to MAT2A inhibition. MTAP, also known as methylthioadenosine phosphorylase, is widely expressed in normal tissue cells. This enzyme can catalyze the conversion of methylthioadenosine (MTA) into 5-methylthioribose-1-phosphate and adenine. This process is also an important link in the methionine salvage pathway in the human body. When MTAP is absent, the metabolic pathway of MTA is inhibited, leading to a large accumulation of MTA in the body and ultimately increasing the sensitivity of cancer cells to MAT2A inhibition.

[0006] The gene encoding human MTAP is located in the chromosomal region 9p21 (chr9p21). The frequency of homozygous deletion of MTAP in all tumors is about 15%, and the deletion frequencies vary in different tumors. Tumor types with relatively high deletion frequencies include glioma, mesothelioma, melanoma, gastric cancer, esophageal cancer, bladder cancer, pancreatic cancer, non-small cell lung cancer, astrocytoma, osteosarcoma, head and neck cancer, myxoid chondrosarcoma, ovarian cancer, endometrial cancer, breast cancer, soft tissue sarcoma, non-Hodgkin lymphoma, etc.

[0007] The chromosomal region 9p21 in humans not only contains the gene encoding MTAP, but also contains the tumor suppressor genes p16INK4A (also known as CDKN2A) and p15INK4B. In 80%-90% of the tumors with CDKN2A deletion, MTAP is also in the deleted state.

[0008] Given that the expression level of MAT2A is abnormally elevated in multiple types of tumors, including gastric cancer, colon cancer, liver cancer, and pancreatic cancer, etc., and selective inhibition of MAT2A can reduce the proliferative activity of MTAP-deleted cancer cells. Therefore, selective inhibition of MAT2A can be used as an effective tumor treatment method.

[0009] During the development of the preparation process of the MAT2A inhibitor with a lactam structure, it was found that there are problems such as cumbersome preparation process routes, expensive reagents, and pollution, which are not conducive to industrial production and there is a need for improvement. Summary of the Invention

[0010] The compound of formula (A) has excellent MAT2A selective inhibitory activity and can be used for preventing or treating tumors caused by reduced or absent MTAP activity.

[0011]

[0012] After multiple explorations and experiments, the inventors developed a preparation method for the compound of formula (A), and unexpectedly found that the compound shown in formula (II) can effectively solve the above problems in the industrial production process as an intermediate.

[0013] On the one hand, the present invention provides a compound shown in formula (II):

[0014]

[0015] Wherein, X is selected from Cl, Br or I;

[0016] L is a leaving group, and the leaving group is selected from F, Cl, Br, I,

[0017] R 1A protecting group which is H or N, and the protecting group for N is selected from formyl, acetyl, trifluoroacetyl, methoxycarbonyl, ethoxycarbonyl, benzyloxycarbonyl, tert-butoxycarbonyl, allyloxycarbonyl, benzyl, p-methoxybenzyl, 2-(trimethylsilyl)ethoxymethyl or methoxymethyl;

[0018] R 2 、R 3 、R 4 are independently selected from the following groups which are optionally substituted by R a : C 1 -C 6 alkyl, C 3 -C 6 cycloalkyl, 3- to 10-membered heterocyclic group, phenyl, 5- to 10-membered heteroaryl;

[0019] Each R a is selected from F, Cl, Br, I, CN, =O, C 1 -C 3 alkyl, OH, NH 2 、NH(C 1 -C 3 alkyl) or N(C 1 -C 3 alkyl) 2 。

[0020] In some embodiments, X is selected from Br.

[0021] In some embodiments, L is selected from F, Br, I,

[0022] In some embodiments, R 2 、R 3 、R 4 are independently selected from C 1 -C 6 alkyl or phenyl, and the C 1 -C 6 alkyl or phenyl is optionally substituted by R a 。

[0023] In some embodiments, R 2 、R 3 、R 4 are independently selected from methyl or phenyl, and the methyl or phenyl is optionally substituted by R a 。

[0024] In some embodiments, R a is selected from F, Cl, Br, I, CN, =O, C 1 -C 3 alkyl, NH 2 、NH(C1 -C 3 alkyl) or N(C 1 -C 3 alkyl) 2 。

[0025] In some embodiments, R a is selected from F or C 1 -C 3 alkyl.

[0026] In some embodiments, R a is selected from F or methyl.

[0027] In some embodiments, L is selected from F, Br, I,

[0028] In some embodiments, L is selected from F, Br or I.

[0029] In some embodiments, L is selected from F or Br.

[0030] In some embodiments, L is selected from F.

[0031] In some embodiments, R 1 is selected from H, tert-butoxycarbonyl, benzyl, 2-(trimethylsilyl)ethoxymethyl or methoxymethyl.

[0032] In some embodiments, R 1 is selected from H.

[0033] In some embodiments, the compound represented by formula (II) is selected from the compounds represented by formula (IIa):

[0034]

[0035] wherein X and L are as defined above.

[0036] In some embodiments, the compound represented by formula (II) is selected from the compounds represented by formula (IIb):

[0037]

[0038] wherein X is as defined above.

[0039] In some embodiments, the compound represented by formula (II) is selected from compound of formula (IIc) compound of formula (IId) compound of formula (IIe) or compound of formula (IIf)

[0040] In some embodiments, the compound represented by formula (II) is selected from the compounds represented by formula (IIc):

[0041]

[0042] Furthermore, the present invention also provides salts of the above-mentioned compound of formula (II), and the salts are acid addition salts formed at any salt-forming site in the compound of formula (II).

[0043] Furthermore, the present invention also provides salts of the above-mentioned compound of formula (II), and the salts are base addition salts formed at any salt-forming site in the compound of formula (II).

[0044] According to the present invention, the acid addition salts are selected from inorganic acid addition salts or organic acid addition salts.

[0045] According to the present invention, the base addition salts are selected from inorganic base addition salts or organic base addition salts.

[0046] Furthermore, the present invention also provides a preparation method of the above-mentioned compound of formula (II), and the preparation method includes step 3:

[0047]

[0048] (Step 3)

[0049] wherein X, L, R 1 are as defined above.

[0050] In some embodiments, R 1 is selected from H.

[0051] In some embodiments, X is selected from Cl, Br or I, L is selected from F or Br, and R 1 is selected from H.

[0052] In some embodiments, X is selected from Br, L is selected from F, and R 1 is selected from H.

[0053] In some embodiments, the halogenating agent in step 3 is selected from a chlorinating agent, a brominating agent or an iodinating agent.

[0054] In some embodiments, the halogenating agent in step 3 is selected from N-chlorosuccinimide, N-bromosuccinimide, N-iodosuccinimide, bromine, iodine, dibromohydantoin, pyridinium tribromide, sodium bromate, 5,5-dibromo-2,2-dimethyl-4,6-dione-1,3-dioxane, 1,3-dibromo-1,3,5-triazine-2,4,6-trione, copper(I) chloride or copper(I) iodide.

[0055] In some embodiments, the halogenating reagent described in step 3 is selected from N-chlorosuccinimide, N-bromosuccinimide, N-iodosuccinimide, bromine, iodine, dibromohydantoin, pyridinium tribromide or sodium bromate.

[0056] In some embodiments, the halogenating reagent described in step 3 is selected from N-chlorosuccinimide, N-bromosuccinimide, bromine, dibromohydantoin, pyridinium tribromide or sodium bromate.

[0057] In some embodiments, the halogenating reagent described in step 3 is selected from N-bromosuccinimide, bromine, dibromohydantoin, pyridinium tribromide or sodium bromate.

[0058] In some embodiments, the molar ratio of the compound of formula (I) to the halogenating reagent in step 3 is 2:1 to 1:2.

[0059] In some embodiments, the reaction solvent in step 3 is selected from one or more of amide solvents, carboxylic acid solvents, alcohol solvents, ester solvents, nitrile solvents, halogenated alkane solvents or ether solvents.

[0060] In some embodiments, the reaction solvent in step 3 is selected from one or more of acetic acid, N,N-dimethylformamide, N,N-dimethylacetamide, N,N-diethylformamide, acetonitrile or carbon tetrachloride.

[0061] In some embodiments, the reaction temperature in step 3 is 0°C - 130°C.

[0062] In some embodiments, the reaction temperature in step 3 is 50°C - 110°C.

[0063] In some embodiments, the reaction time in step 3 is 1 - 24 hours.

[0064] In some embodiments, the reaction time in step 3 is 1 - 10 hours.

[0065] In some embodiments, when the leaving group L of the compound of formula (I) in step 3 is selected from Cl, then X is not Br.

[0066] Furthermore, the present invention also provides a preparation method of the compound of formula (IIa), and the preparation method of the compound of formula (IIa) comprises step 1b, step 2b and step 3b:

[0067] (Step 1b):

[0068] (Step 2b):

[0069] (Step 3b):

[0070] Wherein, X and L are defined as above.

[0071] Furthermore, the present invention provides a method for preparing a compound of formula (IIc), and the method for preparing the compound of formula (IIc) comprises step 1c, step 2c, and step 3c:

[0072]

[0073] In some embodiments, step 1c further includes the step of adding an acidic reagent.

[0074] In some embodiments, the acidic reagent added in step 1c is sulfuric acid.

[0075] In some embodiments, the reaction temperature of step 1c is -5°C to 130°C.

[0076] In some embodiments, the reaction temperature of step 1c is 0°C to 110°C.

[0077] In some embodiments, the reaction time of step 1c is 1 to 24 hours.

[0078] In some embodiments, the reaction time of step 1c is 1 to 5 hours.

[0079] In some embodiments, step 2c further includes the step of adding a fluorinating reagent and a nitrite.

[0080] In some embodiments, the fluorinating reagent added in step 2c is pyridine hydrofluoride or fluoboric acid.

[0081] In some embodiments, the fluorinating reagent added in step 2c is pyridine hydrofluoride.

[0082] In some embodiments, the nitrite added in step 2c is sodium nitrite.

[0083] In some embodiments, the reaction temperature of step 2c is -15°C to 30°C.

[0084] In some embodiments, the reaction temperature of step 2c is -5°C to 10°C.

[0085] In some embodiments, the reaction temperature of step 2c is 0°C to 10°C.

[0086] In some embodiments, the reaction time of step 2c is 5 minutes to 3 hours.

[0087] In some embodiments, the reaction time of step 2c is 5 minutes to 1 hour.

[0088] In some embodiments, step 3c further includes the step of adding a brominating reagent.

[0089] In some embodiments, the brominating reagent added in step 3c is N-bromosuccinimide or bromine.

[0090] In some embodiments, the reaction solvent for step 3c is selected from one or more of amide solvents, carboxylic acid solvents, alcohol solvents, ester solvents, nitrile solvents, halogenated alkane solvents, or ether solvents.

[0091] In some embodiments, the reaction solvent for step 3c is selected from one or more of acetic acid, N,N-dimethylformamide, N,N-dimethylacetamide, N,N-diethylformamide, acetonitrile, or carbon tetrachloride.

[0092] In some embodiments, the reaction solvent for step 3c is N,N-dimethylformamide.

[0093] In some embodiments, the reaction solvent for step 3c is acetic acid.

[0094] In some embodiments, the reaction temperature for step 3c is 0°C - 130°C.

[0095] In some embodiments, the reaction temperature for step 3c is 50°C - 100°C.

[0096] In some embodiments, the reaction temperature for step 3c is 60°C - 90°C.

[0097] In some embodiments, the reaction temperature for step 3c is 75°C - 80°C.

[0098] In some embodiments, the reaction time for step 3c is 1 - 24 hours.

[0099] In some embodiments, the reaction time for step 3c is 1 - 10 hours.

[0100] In some embodiments, the reaction time for step 3c is 1 - 3 hours.

[0101] In some embodiments, the reaction time for step 3c is 2 hours.

[0102] Furthermore, the present invention provides a method for preparing a compound of formula (IId), and the method for preparing the compound of formula (IId) includes step 2d and step 3d:

[0103]

[0104] In some embodiments, step 2d further includes the step of adding a chlorinating reagent and nitrite.

[0105] In some embodiments, the chlorinating reagent added in step 2d is cuprous chloride and HCl.

[0106] In some embodiments, the nitrite added in step 2d is sodium nitrite, potassium nitrite, methyl nitrite, ethyl nitrite, n-butyl nitrite, tert-butyl nitrite or isoamyl nitrite.

[0107] In some embodiments, the nitrite added in step 2d is sodium nitrite.

[0108] In some embodiments, the reaction temperature of step 2d is -15°C to 50°C.

[0109] In some embodiments, the reaction temperature of step 2d is -5°C to 25°C.

[0110] In some embodiments, the reaction temperature of step 2d is 0°C to 25°C.

[0111] In some embodiments, step 3d further includes the step of adding a brominating reagent.

[0112] In some embodiments, the brominating reagent added in step 3d is N-bromosuccinimide or bromine.

[0113] In some embodiments, the brominating reagent added in step 3d is bromine.

[0114] In some embodiments, the reaction solvent of step 3d is selected from one or more of amide solvents, carboxylic acid solvents, alcohol solvents, ester solvents, nitrile solvents, halogenated alkane solvents or ether solvents.

[0115] In some embodiments, the reaction solvent of step 3d is selected from one or more of acetic acid, N,N-dimethylformamide, N,N-dimethylacetamide, N,N-diethylformamide, acetonitrile or carbon tetrachloride.

[0116] In some embodiments, the reaction solvent of step 3d is acetic acid.

[0117] In some embodiments, the molar ratio of the compound of (Id) in step 3d to the brominating reagent is 1:2 to 5:1.

[0118] In some embodiments, the reaction temperature of step 3d is 0°C to 130°C.

[0119] In some embodiments, the reaction temperature of step 3d is 50°C to 120°C.

[0120] In some embodiments, the reaction temperature of step 3d is 110°C.

[0121] In some embodiments, the reaction time of step 3d is 1 - 24 hours.

[0122] In some embodiments, the reaction time of step 3d is 1 - 10 hours.

[0123] In some embodiments, the reaction time of step 3d is 3 hours.

[0124] Furthermore, the present invention provides a method for preparing a compound of formula (A), the method for preparing the compound of formula (A) comprising step 4a, step 4b, step 5a - 1 and step 6a - 1, or comprising step 4a, step 4b, step 5a - 2 and step 6a - 2:

[0125] (Step 4a):

[0126] (Step 4b):

[0127] (Step 5a - 1):

[0128] (Step 6a - 1):

[0129] (Step 5a - 2):

[0130] (Step 6a - 2):

[0131] wherein, X, L are defined as above, R 1 is H or a protecting group defined as above, wherein, when R 1 is H, the compound of formula (III) is the same as the compound of formula (IIIa), and in this case, the method for preparing the compound of formula (A) does not include step 4b.

[0132] In some embodiments, the compound of formula (II) is the compound of formula (IIa).

[0133] In some embodiments, the compound of formula (II) is the compound of formula (IIc).

[0134] In some embodiments, step 4a further comprises the step of adding sodium ethoxide.

[0135] In some embodiments, the reaction solvent for step 4a is selected from ethanol.

[0136] In some embodiments, the reaction temperature for step 4a is 0°C - 130°C.

[0137] In some embodiments, the reaction temperature in step 4a is 60°C - 90°C.

[0138] In some embodiments, the reaction temperature in step 4a is 70°C - 80°C.

[0139] In some embodiments, the reaction time in step 4a is 1 - 24 hours.

[0140] In some embodiments, the reaction time in step 4a is 2 - 10 hours.

[0141] In some embodiments, the reaction time in step 4a is 2.5 hours.

[0142] In some embodiments, the compound of formula (II) is the compound of formula (IId).

[0143] In some embodiments, step 4a further comprises the step of adding sodium ethoxide.

[0144] In some embodiments, the reaction solvent in step 4a is selected from ethanol.

[0145] In some embodiments, the reaction temperature in step 4a is 0°C - 130°C.

[0146] In some embodiments, the reaction temperature in step 4a is 80 - 120°C.

[0147] In some embodiments, the reaction temperature in step 4a is 105°C.

[0148] In some embodiments, the reaction time in step 4a is 1 - 24 hours.

[0149] In some embodiments, the reaction time in step 4a is 16 hours.

[0150] In some embodiments, step 5a - 1 is as follows:

[0151]

[0152] In some embodiments, step 5a - 1 further comprises the step of adding a metal catalyst.

[0153] In some embodiments, step 5a - 1 further comprises the step of adding a basic reagent.

[0154] In some embodiments, the metal catalyst is a palladium - based catalyst.

[0155] In some embodiments, the metal catalyst is dichloro[1,1'-bis(di - tert - butylphosphino)ferrocene]palladium(II) (Pd(dtbpf)Cl2 )。

[0156] In some embodiments, the basic reagent is potassium carbonate, sodium carbonate or potassium phosphate.

[0157] In some embodiments, the basic reagent is potassium phosphate.

[0158] In some embodiments, the reaction solvent in step 5a-1 is selected from one or more of water, amide solvents, sulfone solvents, nitrile solvents, halogenated alkane solvents or ether solvents.

[0159] In some embodiments, the reaction solvent in step 5a-1 is selected from one or more of water, 1,4-dioxane, N,N-dimethylformamide, dimethyl sulfoxide, acetonitrile or carbon tetrachloride.

[0160] In some embodiments, the reaction solvent in step 5a-1 is a mixed solvent of water and 1,4-dioxane.

[0161] In some embodiments, the reaction temperature in step 5a-1 is 60°C - 130°C.

[0162] In some embodiments, the reaction temperature in step 5a-1 is 80°C - 120°C.

[0163] In some embodiments, the reaction temperature in step 5a-1 is 100°C.

[0164] In some embodiments, the reaction time in step 5a-1 is 1 - 24 hours.

[0165] In some embodiments, the reaction time in step 5a-1 is 2 - 10 hours.

[0166] In some embodiments, the reaction time in step 5a-1 is 3 hours.

[0167] In some embodiments, step 6a-1 is:

[0168]

[0169] In some embodiments, step 6a-1 is:

[0170]

[0171] In some embodiments, step 6a-1 further includes the step of adding a metal catalyst.

[0172] In some embodiments, step 6a-1 further includes the step of adding a basic reagent.

[0173] In some embodiments, step 6a-1 further comprises the step of adding a ligand of the metal catalyst.

[0174] In some embodiments, the metal catalyst is copper(I) iodide.

[0175] In some embodiments, the base is potassium carbonate, sodium carbonate or potassium phosphate.

[0176] In some embodiments, the base is potassium carbonate.

[0177] In some embodiments, the ligand of the metal catalyst is N,N-dimethylglycine.

[0178] In some embodiments, the reaction solvent for step 6a-1 is selected from one or more of water, amide solvents, sulfone solvents, nitrile solvents, or ether solvents.

[0179] In some embodiments, the reaction solvent for step 6a-1 is selected from one or more of 1,4-dioxane, N,N-dimethylformamide, dimethyl sulfoxide or acetonitrile.

[0180] In some embodiments, the reaction solvent for step 6a-1 is selected from dimethyl sulfoxide.

[0181] In some embodiments, the reaction temperature for step 6a-1 is 60°C - 150°C.

[0182] In some embodiments, the reaction temperature for step 6a-1 is 80°C - 130°C.

[0183] In some embodiments, the reaction temperature for step 6a-1 is 120°C.

[0184] In some embodiments, the reaction time for step 6a-1 is 1 - 24 hours.

[0185] In some embodiments, the reaction time for step 6a-1 is 6 - 20 hours.

[0186] In some embodiments, the reaction time for step 6a-1 is 16 hours.

[0187] In some embodiments, step 5a-2 is as follows:

[0188]

[0189] In some embodiments, step 5a-2 further comprises the step of adding a metal catalyst.

[0190] In some embodiments, step 5a-2 further comprises the step of adding a base.

[0191] In some embodiments, the metal catalyst is a copper-based catalyst.

[0192] In some embodiments, the metal catalyst is copper acetate, copper trifluoromethanesulfonate, copper trifluoroacetate, copper bromide, copper chloride, copper sulfate, cuprous chloride, bis(neopentanoyl oxy) copper, copper acetylacetonate, tetraethyl cyanocuprate hexafluorophosphate, copper oxide, copper sulfide or copper powder.

[0193] In some embodiments, the metal catalyst is copper acetate.

[0194] In some embodiments, the basic reagent is pyridine.

[0195] In some embodiments, the reaction solvent in step 5a-2 is selected from one or more of water, amide solvents, sulfone solvents, nitrile solvents, haloalkyl solvents or ether solvents.

[0196] In some embodiments, the reaction solvent in step 5a-2 is selected from one or more of dichloromethane, chloroform, 1,4-dioxane, N,N-dimethylformamide, dimethyl sulfoxide or acetonitrile.

[0197] In some embodiments, the reaction solvent in step 5a-2 is selected from dichloromethane.

[0198] In some embodiments, the reaction temperature in step 5a-2 is 10°C - 120°C.

[0199] In some embodiments, the reaction temperature in step 5a-2 is 20°C - 10°C.

[0200] In some embodiments, the reaction temperature in step 5a-2 is 40°C.

[0201] In some embodiments, the reaction time in step 5a-2 is 1 - 24 hours.

[0202] In some embodiments, the reaction time in step 5a-2 is 6 - 20 hours.

[0203] In some embodiments, the reaction time in step 5a-2 is 16 hours.

[0204] In some embodiments, step 6a-2 is as follows:

[0205]

[0206] In some embodiments, step 6a-2 further includes the step of adding a metal catalyst.

[0207] In some embodiments, step 6a-2 further comprises the step of adding a basic reagent.

[0208] In some embodiments, the metal catalyst is a palladium-based catalyst.

[0209] In some embodiments, the metal catalyst is Pd(dtbpf)Cl 2 (dichloro[1,1'-bis(di-tert-butylphosphino)ferrocenepalladium(II)).

[0210] In some embodiments, the basic reagent is potassium carbonate, sodium carbonate, cesium carbonate or potassium phosphate.

[0211] In some embodiments, the basic reagent is cesium carbonate.

[0212] In some embodiments, the reaction solvent for step 6a-2 is selected from one or more of water, amide solvents, sulfone solvents, nitrile solvents, halogenated alkane solvents or ether solvents.

[0213] In some embodiments, the reaction solvent for step 6a-2 is selected from one or more of water, 1,4-dioxane, N,N-dimethylformamide, dimethyl sulfoxide, acetonitrile or carbon tetrachloride.

[0214] In some embodiments, the reaction solvent for step 6a-2 is a mixed solvent of water and 1,4-dioxane.

[0215] In some embodiments, the reaction temperature for step 6a-2 is 60°C - 150°C.

[0216] In some embodiments, the reaction temperature for step 6a-2 is 70°C - 120°C.

[0217] In some embodiments, the reaction temperature for step 6a-2 is 80 - 100°C.

[0218] In some embodiments, the reaction temperature for step 6a-2 is 90°C.

[0219] In some embodiments, the reaction time for step 6a-2 is 1 - 24 hours.

[0220] In some embodiments, the reaction time for step 6a-2 is 6 - 20 hours.

[0221] In some embodiments, the reaction time for step 6a-2 is 15 hours.

[0222] Furthermore, the present invention provides a method for preparing a compound of formula (A), and the method for preparing the compound of formula (A) comprises step 4a, step 5a-1 and step 6a-1:

[0223]

[0224] In some embodiments, Step 4a, Step 5a-1, and Step 6a-1 are as defined above.

[0225] Furthermore, the present invention provides a method for preparing a compound of formula (A), the method for preparing the compound of formula (A) comprising Step 4a, Step 5a-2, and Step 6a-2:

[0226]

[0227] In some embodiments, Step 4a, Step 5a-2, and Step 6a-2 are as defined above.

[0228] Furthermore, the present invention provides a method for preparing a compound of formula (A), the method for preparing the compound of formula (A) comprising Step 4a, Step 5a-2, and Step 6a-2:

[0229]

[0230] In some embodiments, Step 4a, Step 5a-2, and Step 6a-2 are as defined above.

[0231] Furthermore, the present invention also relates to the use of a compound of formula (II) or a salt thereof as an intermediate in the preparation of a compound of formula (A),

[0232]

[0233] wherein R 1 , X, and L are as defined above.

[0234] In some embodiments, the compound of formula (II) is selected from compounds of formula (IIa)

[0235] In some embodiments, the compound of formula (II) is selected from compounds of formula (IIb)

[0236] In some embodiments, the compound of formula (II) is selected from compounds of formula (IIc) Compound of formula (IId) Compound of formula (IIe) or compound of formula (IIf)

[0237] In some embodiments, the present invention relates to the use of a compound of formula (IIa) or a salt thereof as an intermediate in the preparation of a compound of formula (A),

[0238]

[0239] Wherein, X and L are defined as above.

[0240] In some embodiments, the present invention relates to the use of a compound of formula (IIb) or a salt thereof as an intermediate in the preparation of a compound of formula (A),

[0241]

[0242] Wherein, X is defined as above.

[0243] In some embodiments, the present invention relates to the use of a compound of formula (IIc) or a salt thereof as an intermediate in the preparation of a compound of formula (A),

[0244]

[0245] In some embodiments, the present invention relates to the use of a compound of formula (Ic) or a salt thereof as an intermediate in the preparation of a compound of formula (A),

[0246]

[0247] The preparation method of the present invention can effectively prepare the compound of formula (A), and has the advantages of cheap and easily available raw materials, economical and environmental protection of the synthesis route, etc., and can meet the requirements of industrial production.

[0248] Term Definitions and Explanations

[0249] Unless otherwise specified, the definitions of groups and terms recorded in the specification and claims of the present application, including their definitions as examples, exemplary definitions, preferred definitions, definitions recorded in tables, definitions of specific compounds in examples, etc., can be combined and combined with each other arbitrarily. The group definitions and compound structures after such combination and combination should fall within the scope recorded in the specification of the present application.

[0250] The term "pharmaceutically acceptable salt" refers to a non-toxic acid or base salt acceptable in pharmacy, including salts of inorganic acids and bases, organic acids and bases.

[0251] The compounds of the present invention may have asymmetric carbon atoms (optical centers) or double bonds. Racemates, diastereoisomers, geometric isomers and individual isomers are all included within the scope of the present invention.

[0252] The graphical representations of racemates or enantiomerically pure compounds in this text are from Maehr, J. Chem. Ed. 1985, 62: 114 - 120. Unless otherwise specified, the absolute configuration of a stereocenter is represented by a wedge bond and a dashed bond. When the compounds described herein contain an olefinic double bond or other geometrically asymmetric centers, unless otherwise specified, they include E, Z geometric isomers. Similarly, all tautomeric forms are included within the scope of the present invention.

[0253] 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, diastereoisomers, (D)-isomers, (L)-isomers, and their racemic mixtures and other mixtures, such as enantiomer- or diastereoisomer-enriched mixtures, all of which mixtures are within the scope of the present invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and their mixtures are included within the scope of the present invention.

[0254] The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, provided that the valence of the particular atom is normal and the resulting compound is stable. When the substituent is oxo (i.e., =O), it means that two hydrogen atoms are replaced, and oxo does not occur on an aromatic group.

[0255] The term "optionally" or "optionally" means that the subsequent described event or circumstance may or may not occur, and the description includes both the occurrence and non-occurrence of the described event or circumstance. For example, "optionally" substituted by a halogen for an ethyl group means that the ethyl group may be unsubstituted (CH 2 CH 3 ), monosubstituted (such as CH 2 CH 2 F), polysubstituted (such as CHFCH 2 F, CH 2 CHF 2 , etc.) or fully substituted (CF 2 CF 3 ). Those skilled in the art will understand that for any group containing one or more substituents, no substitutions or substitution patterns that are spatially impossible to exist and / or cannot be synthesized will be introduced.

[0256] The term "C 1 -C 6"Alkyl" shall be understood to mean a straight-chain or branched-chain saturated monovalent hydrocarbon group having 1, 2, 3, 4, 5 or 6 carbon atoms. The alkyl groups are, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl or 1,2-dimethylbutyl, etc. The term "C 1 -C 3 alkyl" shall be understood to mean methyl, ethyl, propyl or isopropyl.

[0257] The term "C 3 -C 6 cycloalkyl" shall be understood to mean a saturated monovalent hydrocarbon ring having 3 to 6 carbon atoms. It includes but is not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl.

[0258] The term "3- to 10-membered heterocyclic group" means a saturated or partially saturated monovalent monocyclic, fused-ring, spiro-ring or bridged-ring which contains 1 to 5, preferably 1 to 3 heteroatoms selected from N, O, B and S. In particular, the heterocyclic group may include but is not limited to: 4-membered rings such as azetidinyl, oxetanyl; 5-membered rings such as tetrahydrofuranyl, dioxolanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl, 1,3,2-dioxaborolanyl; or 6-membered rings such as tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl or trithianyl; or partially saturated 6-membered rings such as tetrahydropyridinyl; or 7-membered rings such as diazepanyl. Optionally, the heterocyclic group may be benzo-fused. The heterocyclic group may be bicyclic, for example but not limited to 5,5-membered rings such as hexahydrocyclopenta[c]pyrrol-2(1H)-yl ring, or 5,6-membered bicyclic rings such as hexahydropyrrolo[1,2-a]pyrazin-2(1H)-yl ring. Rings containing nitrogen atoms may be partially unsaturated, i.e. it may contain one or more double bonds, such as but not limited to 2,5-dihydro-1H-pyrrolyl, 4H-[1,3,4]thiadiazinyl, 4,5-dihydrooxazolyl or 4H-[1,4]thiazinyl, or, it may be benzo-fused, such as but not limited to dihydroisoquinolinyl. Optionally, the 3- to 10-membered heterocyclic group may be a "3- to 10-membered heterocycloalkyl", meaning a saturated monovalent monocyclic, fused-ring, spiro-ring or bridged-ring containing 1 to 5 heteroatoms; according to the present invention, the heterocyclic group is non-aromatic.

[0259] The term "5- to 10-membered heteroaryl" should be understood to include a monocyclic, bicyclic or tricyclic aromatic ring system having 5 to 10 ring atoms and containing 1 to 5 heteroatoms independently selected from N, O and S. "5- to 6-membered heteroaryl" refers to a ring system having 5 or 6 ring atoms and containing 1 to 4, preferably 1 to 3 heteroatoms independently selected from N, O and S. In particular, heteroaryl is selected from thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl, etc. and their benzo derivatives, such as benzofuryl, benzothienyl, benzothiazolyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, benzotriazolyl, indazolyl, indolyl, isoindolyl, etc.; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, etc. and their benzo derivatives, such as quinolinyl, quinazolinyl, isoquinolinyl, etc.

[0260] The term "acid addition salt" should be understood to mean a salt formed by the compound with an inorganic acid or an organic acid; inorganic acid salts include, but are not limited to, hydrochloride, hydrobromide, sulfate, nitrate, phosphate, etc.; organic acid salts include, but are not limited to, formate, acetate, 2,2-dichloroacetate, trifluoroacetate, propionate, caproate, caprylate, caprate, undecylenate, glycolate, gluconate, lactate, sebacate, adipate, glutarate, malonate, oxalate, maleate, succinate, fumarate, tartrate, citrate, palmitate, stearate, oleate, cinnamate, laurate, malate, glutamate, pyroglutamate, aspartate, benzoate, methanesulfonate, benzenesulfonate, p-toluenesulfonate, alginate, ascorbate, salicylate, 4-aminosalicylate, naphthalenedisulfonate, etc. These salts can be prepared by methods known in the art.

[0261] The term "base addition salt" should be understood to mean a salt formed by the compound with an inorganic base or an organic base; salts of inorganic bases include, but are not limited to, sodium salt, potassium salt, lithium salt, ammonium salt, calcium salt, magnesium salt, iron salt, zinc salt, copper salt, manganese salt, aluminum salt, etc.; preferred inorganic salts are ammonium salt, sodium salt, potassium salt, calcium salt and magnesium salt. Salts of organic bases include, but are not limited to, the following salts: isopropylamine salt, trimethylamine salt, diethylamine salt, triethylamine salt, tripropylamine salt, ethanolamine salt, diethanolamine salt, triethanolamine salt, lysine salt, arginine salt, histidine salt. These salts can be prepared by methods known in the art.

[0262] The term "ether solvents" includes, but is not limited to, diethyl ether, methyl ethyl ether, dipropyl ether, dibutyl ether, 1,4-dioxane, furan, methylfuran, tetrahydrofuran.

[0263] The term "amide solvents" includes, but is not limited to, N,N-dimethylformamide, N,N-dimethylacetamide, N,N-diethylformamide.

[0264] The term "carboxylic acid solvent" includes, but is not limited to, acetic acid and propionic acid.

[0265] The term "sulfone solvent" includes, but is not limited to, dimethyl sulfoxide, dimethyl sulfone, sulfolane, and 2,4-dimethyl sulfolane.

[0266] The term "ester solvent" includes, but is not limited to, methyl acetate, ethyl acetate, hexyl acetate, and phenyl acetate.

[0267] The term "nitrile solvent" includes, but is not limited to, acetonitrile.

[0268] The term "alcohol solvent" includes, but is not limited to, methanol, ethanol, propanol, isopropanol, butanol, pentanol, decanol, n-dodecanol, cyclopentanol, cyclohexanol, benzyl alcohol, and phenethyl alcohol.

[0269] The term "haloalkane solvent" includes, but is not limited to, chloromethane, dichloromethane, chloroform, carbon tetrachloride, and 1,2-dichloroethane.

[0270] The term "room temperature" or its abbreviation "r.t." refers to 25 ± 5 °C.

[0271] The acidic reagent described in the present invention can be a Lewis acid or a protonic acid, such as an organic acid or an inorganic acid, for example, one, two, or more of formic acid, acetic acid, trifluoroacetic acid, hydrochloric acid, nitric acid, sulfuric acid, aluminum chloride, iron chloride, boron trifluoride, niobium pentachloride, and nitric acid. Detailed Description of the Invention

[0272] The following examples illustrate the technical solutions of the invention in detail, but the protection scope of the present invention includes, but is not limited to, this.

[0273] The structure of the compound is determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). The unit of NMR shift is 10 -6 (ppm). The solvents for NMR measurement are deuterated dimethyl sulfoxide, deuterated chloroform, deuterated methanol, etc., and the internal standard is tetramethylsilane (TMS); "IC 50 " refers to the half inhibitory concentration, which means the concentration when the maximum inhibitory effect reaches half.

[0274] Preparation of Compound (IIc) and Compound of Formula (A) in Example 1

[0275] Synthesis route and specific synthesis steps:

[0276]

[0277] Preparation of S1

[0278]

[0279] Under an ice-water bath, add the reactants 2,6-diaminopyridine (67.5 g, 503.73 mmol, 1.10 eq) and DL-malic acid (50 g, 458.71 mmol, 1.00 eq) to a 2 L three-necked flask. Stir the mixture evenly using mechanical stirring, and then slowly add concentrated sulfuric acid (230 mL). During the addition of concentrated sulfuric acid, control the temperature not to exceed 45 °C. After the addition is complete, stir the reaction solution at 110 °C for 3 h. LCMS detects the completion of the reaction. Cool the reaction solution to room temperature and pour it into ice water (1 L) to quench. Add ammonia water to adjust the pH of the solution to 9, and a solid precipitates. Filter the reaction solution, and wash the filter cake with water twice. Pulverize the filter cake with a mixed solution of H 2 O / MeOH = 1:9. Filter and collect the solid. The obtained solid is dried under vacuum to obtain S1 (89 g of crude product), without purification, and the compound is directly used in the next reaction.

[0280] LC-MS: m / z: ES+[M+H] + = 162.

[0281] 1 H NMR (400 MHz, DMSO-d 6 ) δ 11.85 (s, 1H), 7.67 (dd, J = 8.9, 2.4 Hz, 2H), 6.99 (s, 2H), 6.37 (d, J = 8.5 Hz, 1H), 6.14 (d, J = 9.3 Hz, 1H).

[0282] Preparation of Ic

[0283]

[0284] Under an ice-water bath, dissolve the reactant S1 (60 g, crude product) in 800 mL of HF-Py (65% - 70% wt hydrogen fluoride). Add NaNO 2 (58.78 g, 851.85 mmol) to the above solution in batches. Control the temperature of the reaction solution between 0 and 10 °C during the feeding process. After the feeding is complete, stir the reaction solution at 0 °C for 5 min. LC-MS detects the completion of the reaction. Add ice water (600 ml) to the reaction solution, and a solid precipitates. Filter the above mixture, wash the solid with water twice, collect the solid and dry it under vacuum to obtain Ic (40 g of crude product). Without purification, the compound is directly used in the next reaction.

[0285] LC-MS: m / z: ES+[M+H] + = 165.

[0286] 1 H NMR (400 MHz, DMSO-d 6)δ12.35(s,1H),8.30(t,J=8.2Hz,1H),7.96(d,J=9.5Hz,1H),7.01(dd,J=8.3,2.1Hz,1H),6.55(d,J=9.6Hz,1H).

[0287] Preparation of Compound IIc

[0288]

[0289] Dissolve the reactant Ic (10 g) in DMF (600 mL), and add N-bromosuccinimide (16 g, 89.88 mmol). The reaction mixture is reacted at 75 °C for 2 h, and the reaction is monitored by LCMS until completion. The reaction mixture is cooled to room temperature, diluted with water (200 mL), and extracted with ethyl acetate (200 mL × 3). The combined organic phases are washed with saturated brine (500 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate is concentrated under reduced pressure to dryness. The concentrated residue is triturated and purified with acetonitrile (10 mL). Filter, collect the solid and dry it to obtain Compound IIc (8 g, 33.04 mmol, total yield of 42% for the three steps of step 1c, 2c, and 3c).

[0290] LC-MS: m / z: ES+[M+H] + =243 / 245.

[0291] 1 H NMR(400MHz,DMSO-d 6 )δ12.87(s,1H),8.56(s,1H),8.30(t,J=8.2Hz,1H),7.06(dd,J=8.4,2.0Hz,1H).

[0292] Preparation of IIIc

[0293]

[0294] Dissolve Compound IIc (5.4 g, 22.2 mmol, 1 eq) in absolute ethanol (100 mL), and add sodium ethoxide (6 g, 88.8 mmol, 4 eq). The reaction mixture is stirred at 80 °C for 2.5 h and monitored by LCMS until the reaction is complete. The reaction mixture is concentrated under reduced pressure, water (150 mL) is added, and it is extracted with ethyl acetate (100 mL × 3). The combined organic phases are dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. Compound IIIc (5.7 g, yield 93.3%) is obtained. Without purification, Compound IIIc is directly used in the next reaction.

[0295] 1 H NMR(400MHz,DMSO-d6 ) δ 12.52 (s, 1H), 8.42 (s, 1H), 7.98 (d, J = 8.5 Hz, 1H), 6.69 (d, J = 8.5 Hz, 1H), 4.38 (q, J = 7.0 Hz, 2H), 1.35 (t, J = 7.0 Hz, 3H).

[0296] Preparation of V

[0297]

[0298] Under a nitrogen atmosphere, reactant IIIc (1.65 g, 6.16 mmol, 1.00 eq), 1-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one (1.74 g, 7.39 mmol, 1.20 eq), Pd(dtbpf)Cl 2 (dichloro[1,1'-bis(di-tert-butylphosphino)ferrocene]palladium(II)) (401.4 mg, 0.616 mmol, 0.1 eq) and K 3 PO 4 (2.61 g, 12.31 mmol, 2.00 eq) were dissolved in a mixed solution of dioxane and water (30 mL of dioxane and 8 mL of water). The reaction mixture was stirred at 100 °C for 3 h, and the reaction was monitored by LCMS until completion. The reaction mixture was concentrated, water (50 mL) was added, and the mixture was filtered. The filter cake was dissolved in a mixed solvent of dichloromethane and CH 3 OH (100 mL of dichloromethane, 25 mL of methanol), activated carbon (1.5 g) was added, and the mixture was stirred at 45 °C for 1 h. The mixture was allowed to stand, and the mixture was filtered through diatomaceous earth. The filtrate was concentrated to obtain a solid (1.5 g, crude product). Dichloromethane and CH 3 OH (20 mL of dichloromethane, 0.5 mL of methanol) were added to the above solid for purification by trituration. The mixture was filtered, the solid was collected and dried to obtain compound V (1.04 g, 3.50 mmol, yield 57.8%).

[0299] LC-MS: m / z: ES+ [M + H] + = 298.

[0300] 1 1H NMR (400 MHz, DMSO-d 6)δ12.24(s,1H),8.35(d,J=2.6Hz,1H),8.05(s,1H),7.98(d,J=8.5Hz,1H),7.84(dd,J=9.5,2.6Hz,1H),6.67(d,J=8.5Hz,1H),6.45(d,J=9.5Hz,1H),4.39(q,J=7.0Hz,2H),3.50(s,3H),1.35(t,J=7.1Hz,3H).

[0301] Preparation of Compound A

[0302]

[0303] Under a nitrogen atmosphere, the reactant V (500 mg, 1.68 mmol, 1.00 eq), 1-(difluoromethoxy)-4-iodobenzene (910 mg, 3.37 mmol, 2.0 eq), N,N-dimethylglycine (70.0 mg, 0.68 mmol, 0.4 eq), K 2 CO 3 (470 mg, 3.37 mmol, 2.00 eq) and CuI (130 mg, 0.68 mmol, 0.4 eq) were successively added to DMSO (5 mL). The reaction mixture was stirred at 120 °C for 16 h, and the reaction was monitored by LCMS until completion. The reaction mixture was cooled to room temperature, and water (10 mL) and ammonia water (25 - 28% wt, 1 mL) were added thereto. The mixture was extracted with ethyl acetate (20 mL × 2), and the combined organic phases were washed with saturated brine (10 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue after concentration was triturated and purified with acetonitrile (5 mL) to obtain pure product A (500 mg, 1.14 mmol, yield 67.7%).

[0304] LC-MS: m / z: ES+[M+H] + =440.

[0305] 1 1H NMR (400 MHz, DMSO-d 6 )δ8.34(d,J=2.6Hz,1H),8.21(s,1H),8.10(d,J=8.5Hz,1H),7.90–7.84(m,1H),7.44–7.29(m,5H),6.73(d,J=8.4Hz,1H),6.47(d,J=9.5Hz,1H),4.02–3.91(m,2H),3.49(s,3H),1.08(t,J=7.0Hz,3H).

[0306] Preparation of Compound of Formula (IId) and Compound of Formula (A) in Example 2

[0307] Synthetic Route and Specific Synthetic Steps:

[0308]

[0309] Preparation of Id

[0310]

[0311] Under an ice - water bath, a solution of sodium nitrite (4.29 g, 62.11 mmol, 2 eq) in water (10 mL) was slowly added dropwise to a solution of reactant S1 (5 g, 31.02 mmol, 1 eq) in concentrated hydrochloric acid (20 mL, 34% wt, 90.17 eq) to obtain a diazonium salt solution. Cuprous chloride (6.14 g, 62.04 mmol, 2 eq) was added to concentrated hydrochloric acid (10 mL, 34% wt, 1 eq) to obtain a yellow solution, and this yellow solution was added dropwise to the above - mentioned diazonium salt solution. After the addition was completed, the reaction mixture was stirred at 0 °C for 1 hour and then at room temperature for 16 hours to obtain a dark - brown suspension. After the reaction was completed, the reaction mixture was filtered to obtain a filter cake and a filtrate. The filtrate was extracted with dichloromethane (30 mL * 5). The combined extraction phases were dried over anhydrous sodium sulfate and then concentrated under reduced pressure to dryness to obtain a solid. The solid and the filter cake were combined and purified by column chromatography (40 g silica gel flash column chromatography, fraction gradient: methanol / dichloromethane 0 - 10%) to obtain product Id (1.1 g, 6.07 mmol, yield 19.6%).

[0312] LC - MS: m / z: ES+[M + H] + = 181.

[0313] Preparation of IId

[0314]

[0315] Compound Id (100 mg, 0.56 mmol, 1 eq) was dissolved in 1 mL of acetic acid, and bromine liquid (890 mg, 2.8 mmol, 5 eq) was added dropwise at room temperature. After the addition was completed, the reaction mixture was stirred at 110 °C for 3 hours. Monitored by LC - MS, starting material Id disappeared. Saturated Na 2 SO 3 solution (10 mL) was added to the reaction mixture and stirred for 2 minutes. The above - mentioned mixture was extracted with ethyl acetate (5 mL * 2). The organic phase was washed with saturated sodium carbonate solution (10 mL * 3), washed with saturated brine (10 mL * 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to dryness to obtain solid IId (160 mg, crude product, 65% purity). Without purification, the crude product was directly used for the next reaction.

[0316] LC-MS: m / z: ES+[M+H] + = 305 / 307.

[0317] Preparation of IIIc

[0318]

[0319] Under a nitrogen atmosphere, sodium ethoxide (188 mg, 2.76 mmol) was added to a solution of compound IId (160 mg, crude, 65% purity) in ethanol (2 mL). The reaction mixture was heated to 105 °C and stirred overnight to obtain a yellow suspension. The reaction mixture was concentrated under reduced pressure to dryness, dissolved in water (10 mL), and extracted with ethyl acetate (3 mL × 3). The organic phase was concentrated under reduced pressure to dryness to obtain solid IIIc (120 mg, crude, 39% purity). Without purification, the crude product was directly used for the next reaction.

[0320] LC-MS: m / z: ES+[M+H] + = 269 / 271.

[0321] Preparation of IV

[0322]

[0323] Compound IIIc (120 mg, crude, 39% purity), compound 4-(difluoromethoxy)phenylboronic acid (126 g, 0.67 mmol), copper(II) acetate (120 mg, 0.668 mmol, 1.1 eq), and pyridine (106 mg, 1.34 mmol) were dissolved in dichloromethane (1 mL). Air was bubbled through the reaction mixture, and the reaction was carried out at 40 °C for 16 h. TLC showed that the reaction was complete. Water (1 mL) was added to the reaction mixture, and the mixture was extracted with dichloromethane (3 mL × 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to dryness to obtain a yellow oil. The oil was purified by column chromatography (petroleum ether:ethyl acetate = 10:1), and the product was collected and concentrated to obtain solid compound IV (87 mg, 39% purity).

[0324] LC-MS: m / z: ES+[M+H] + = 411 / 413.

[0325] Preparation of A

[0326]

[0327] Under a nitrogen atmosphere, compound IV (50 mg, 121.95 μmol, 1.00 eq) and 1-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2(1H)-one (42.9 mg, 182.92 μmol, 1.50 eq) were dissolved in a mixed solution of dioxane (0.8 mL) and water (0.4 mL). Cesium carbonate (118.9 mg, 365.85 μmol, 3.00 eq) and Pd(dtbpf)Cl 2 (dichloro[1,1'-bis(di-tert-butylphosphino)ferrocene)palladium(II)) (10 mg, 12.36 μmol, 0.10 eq) were successively added to the solution. The reaction mixture was reacted at 90 °C for 15 h. The reaction was monitored by TLC and was complete. The reaction mixture was cooled to room temperature, filtered, and the filtrate was collected. The filtrate was purified by preparative high-pressure liquid chromatography [YMC-Actus Triart C18 column, 5 μm silica, 30 mm diameter, 150 mm length; using a polarity-decreasing mixture of water (containing 0.05% NH 4 HCO 3 ) and acetonitrile as the eluent; acetonitrile gradient ratio 70% - 82%, elution time 12 min], and the product was collected and lyophilized to obtain solid A (9.10 mg, yield: 17.16%).

[0328] LC-MS: m / z: ES+ [M+H] + = 440.

[0329] 1 1H NMR (400 MHz, DMSO-d 6 ) δ 8.34 (d, J = 2.6 Hz, 1H), 8.21 (s, 1H), 8.10 (d, J = 8.5 Hz, 1H), 7.90–7.85 (m, 1H), 7.44–7.29 (m, 5H), 6.73 (d, J = 8.4 Hz, 1H), 6.47 (d, J = 9.5 Hz, 1H), 4.03–3.94 (m, 2H), 3.49 (s, 3H), 1.08 (t, J = 7.0 Hz, 3H).

[0330] Example 3 Preparation of the compound of formula (IIc)

[0331]

[0332] Compound Ic (500 mg, 3.10 mmol, 1 eq) was added to acetic acid (7 mL), and bromine (641 mg, 4.03 mmol, 1.3 eq) was added dropwise at room temperature. After the addition was complete, the reaction mixture was stirred at 75 °C for 3 h. The reaction was monitored by LC-MS until completion. The reaction mixture was cooled to room temperature, and saturated Na 2 SO 3 solution (10 mL) was added, and the mixture was stirred for 2 min. The above mixture was extracted with ethyl acetate (50 mL × 2). The organic phase was washed with saturated aqueous sodium carbonate solution (30 mL × 3), washed with saturated aqueous NaCl solution (20 mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to dryness to obtain solid IIc (410 mg, crude product).

[0333] LC-MS: m / z: ES+ [M + H] + = 243 / 245.

[0334] 1 H NMR (400 MHz, DMSO-d 6 ) δ 12.85 (s, 1H), 8.56 (s, 1H), 8.29 (td, J = 8.2, 1.9 Hz, 1H), 7.06 (dd, J = 8.4, 2.0 Hz, 1H).

[0335] Example 4 Preparation of Compound of Formula (IIe)

[0336]

[0337] Compound Ic (161 mg, 1.00 mmol, 1 eq) was added to DMF (3 mL). NIS (1.73 g, 10.00 mmol, 10 eq) was added at room temperature, and the reaction mixture was stirred at 80 °C for 18 h. The reaction was monitored by LC-MS until completion. The reaction mixture was cooled to room temperature, and 3 mL of saturated Na 2 SO 3 solution was added, and the mixture was stirred for 2 min. The above mixture was extracted with ethyl acetate (50 mL × 2). The organic phase was washed with saturated aqueous NaCl solution (20 mL × 1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to dryness and purified by column chromatography (petroleum ether:ethyl acetate = 1:1) to obtain compound IIe (118 mg, yield 40%).

[0338] LC-MS: m / z: ES+ [M + H] + = 291.

[0339] 1 H NMR (400 MHz, DMSO-d 6)δ 12.73 (s, 1H), 8.76 (s, 1H), 8.27 (t, J = 8.2 Hz, 1H), 7.03 (dd, J = 8.4, 1.9 Hz, 1H).

[0340] Example 5 Preparation of Compound of Formula (IIf)

[0341]

[0342] Compound Ic (161 mg, 1.00 mmol, 1 eq) was added to DMF (3 mL). NCS (133 mg, 1.00 mmol, 1 eq) was added at room temperature, and the reaction mixture was stirred at 80 °C for 18 h. The reaction was monitored by LC-MS until completion. The reaction mixture was cooled to room temperature, diluted with water (20 mL), and extracted with ethyl acetate (5 mL × 3). The combined organic phases were washed with saturated brine (20 mL × 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to dryness. The concentrated residue was triturated and purified with a mixed solvent of ethyl acetate and petroleum ether (3 mL petroleum ether + 3 mL ethyl acetate), filtered by suction, the filter cake was collected, and the filter cake was dried in vacuo to obtain solid IIf (130 mg, yield 66%).

[0343] LC-MS: m / z: ES+ [M + H] + = 199 / 201.

[0344] 1 H NMR (400 MHz, DMSO-d 6 )δ 12.93 (s, 1H), 8.37 (s, 1H), 8.30 (t, J = 8.2 Hz, 1H), 7.08 (dd, J = 8.4, 2.0 Hz, 1H).

[0345] Test Examples of Biological Activity and Related Properties of Compound of Formula (A)

[0346] Test Example 1, Biochemical Test

[0347] Brief Introduction of Test Principle: L-Methionine and ATP can be converted into SAM, inorganic phosphate and inorganic diphosphate under the catalysis of MAT2A enzyme. By adding a color reagent, such as ammonium molybdate, etc., to the enzyme reaction mixture, the content of inorganic phosphate in the sample can be quantitatively detected, and then the enzyme activity of MAT2A can be reflected.

[0348] Materials: MAT2A screening kit was purchased from BPS bioscience (USA); 384-well plates were purchased from Corning (USA).

[0349] 1. MAT2a protein (Beijing Kanglonghuacheng New Drug Technology Co., Ltd.);

[0350] 2. L-Methionine (Sigma#M9625-5G)

[0351] 3. ATP (Sigma#A7699-1G)

[0352] 4. KCl (Sigma#60142-500ML-F)

[0353] 5. Tris (Sigma#T2663-1L)

[0354] 6. MgCl 2 (Sigma#M1028)

[0355] 7. EDTA (Invitrogen#AM9260G)

[0356] 8. BSA (Sangon Biotech#A500023-0100)

[0357] 9. PiColorLock (abcam#ab270004)

[0358] Detection method: Dissolve the compound in DMSO, dilute the compound to a final concentration of 10 μM using Echo, perform a 3-fold dilution, and transfer 80 nL to a 384-well plate. Prepare the experimental buffer (50 mM Tris, 50 mM KCl, 15 mM MgCl 2 , 100 μM EDTA, 0.005% BSA). Dilute MAT2a protein with the experimental buffer (final concentration is 4 μg / mL). Add 40 μL of 2X MAT2A solution to the 384-well plate and incubate at room temperature for 60 minutes. Dilute L-methionine and ATP with the experimental buffer (final concentration of L-methionine is 200 μM, final concentration of ATP is 400 μM). Add 40 μL of 2X L-methionine and ATP solution to initiate the reaction and incubate at room temperature for 60 minutes. Add the detection reagent and detect the signal value after incubating at room temperature for 15 minutes.

[0359] Data analysis:

[0360] Calculate % Compound inhibition and fit to obtain the IC 50

[0361] % Compound inhibition = 1 - 100% * (Signal - Bottom) / (Top - Bottom)

[0362] Experimental results:

[0363] Under the conditions of this experiment, the inhibitory effect of the compound to be tested on MAT2A can be represented by the IC 50 value of the inhibition of the level of phosphate production during the enzymatic reaction. The MAT2A inhibitory activity of the compound to be tested is shown in Table 1 specifically.

[0364] Table 1

[0365]

[0366] Test Example 2: Detection of intracellular SAM level

[0367] Brief introduction of the test principle: After co-incubating the MAT2A inhibitor to be tested with cancer cells for a period of time, the cells are lysed with a termination reagent to quench the MAT2A enzyme activity. The MAT2A catalytic product SAM in the cell lysate is quantitatively determined by LC-MS / MS, and then the activity of MAT2A in the cells is reflected.

[0368] Materials and cells: HCT116 MTAP - / - The cells were purchased from Kangyuan Bochuang; fetal bovine serum, McCoy's 5a medium and penicillin-streptomycin were purchased from Gibco (USA), 96-well plates were purchased from Corning (USA), and PBS was purchased from Cytiva (USA).

[0369] Cell culture: HCT116 MTAP - / - The cells were cultured in McCoy's 5a culture medium containing 10% fetal bovine serum + 1% penicillin-streptomycin at 37 °C and 5% CO 2 conditions. Cells in the logarithmic growth phase can be used for the experiment.

[0370] LC-MS / MS detection: Use LC-MS / MS to detect the effect of the compound on the SAM production level in the HCT116 MTAP - / - cell line. Adjust the cell concentration and inoculate into 96-well plates, and culture at 37 °C and 5% CO 2 conditions for 24 hours. After adding the compound, continue to culture at 37 °C and 5% CO 2 conditions for 6 hours. Add the termination solution to lyse the cells. After the lysate is processed, it is injected and analyzed by LC-MS / MS to determine the SAM concentration.

[0371] Data analysis:

[0372] Calculate %Compound inhibition and fit to obtain the IC of the compound 50

[0373] % Compound inhibition = 1 - 100% * (Signal - Bottom) / (Top - Bottom)

[0374] Experimental results:

[0375] Under the conditions of this experiment, the inhibitory activity of the compound to be tested against MAT2A enzyme in HCT116 MTAP - / - cell line is expressed as the IC 50 value of the inhibition of SAM generation in cells by the compound to be tested. The specific inhibitory activity of the compound in formula (A) against SAM generation in cells is shown in Table 2.

[0376] Table 2

[0377]

[0378] Test Example 3: Inhibitory Test on the Proliferation of Human Colon Cancer HCT116 Cells

[0379] Brief Introduction of Test Principle: After co - incubating the MAT2A inhibitor to be tested with cancer cells for a period of time, a cell proliferation counting method based on ATP content is used to measure the effect of the compound to be tested on cell proliferation.

[0380] Materials and Cells: HCT116 MTAP + / + cells and HCT116 MTAP - / - cells were purchased from Kangyuan Bochuang; fetal bovine serum, McCoy's 5a medium, and penicillin - streptomycin were purchased from Gibco (USA), 96 - well plates were purchased from Corning (USA), and Cell - Titer Glo reagent was purchased from Promega (USA).

[0381] Cell Culture: HCT116 MTAP + / + cells and HCT116 MTAP - / - cells were both cultured in McCoy's 5a culture medium containing 10% fetal bovine serum + 1% penicillin - streptomycin at 37°C and 5% CO 2 conditions. Cells in the logarithmic growth phase can be used for the experiment.

[0382] Detection of Cell Proliferation Activity: Use Cell - Titer Glo reagent to detect the inhibitory activity of the compound against the proliferation of HCT116 MTAP + / + and HCT116MTAP - / - two cell lines. Adjust the cell concentration and inoculate into 96 - well plates, and culture at 37°C and 5% CO 2 conditions for 24 hours. After adding the compound, culture at 37°C and 5% CO 2Continue culturing for 6 days under the conditions. Add Cell-Titer Glo reagent to detect cell viability.

[0383] Data analysis:

[0384] Calculate the % Compound inhibition and fit to obtain the IC of the compound 50

[0385] % Compound inhibition = 1 - 100% * (Signal - Bottom) / (Top - Bottom)

[0386] HCT116 MTAP - / - The cells were obtained by site-directed knockout of wild-type HCT116 (HCT116 WT, CT116MTAP + / + ) cells through gene knockout means and do not express MTAP protein. Since MAT2A inhibition and MTAP deletion can produce synthetic lethality, resulting in the death of tumor cells. By testing the anti-proliferative activity of MAT2A inhibitors against HCT116 MTAP - / - cells and HCT116 WT, the activity and selectivity of MAT2A inhibitors can be evaluated.

[0387] Experimental results:

[0388] Under the experimental conditions, the test compound showed strong anti-proliferative activity against HCT116 MTAP - / - cells and showed a certain selectivity compared to HCT116 MTAP + / + cells. The specific anti-cell proliferation activity of the test compound is shown in Table 3.

[0389] Table 3

[0390]

Claims

1. Process for preparing a compound of formula (A), characterized in that, the process for preparing the compound of formula (A) comprises step 4a, step 4b, step 5a-1 and step 6a-1, or comprises step 4a, step 4b, step 5a-2 and step 6a-2: (Step 4a): (Step 4b): (Step 5a-1): (Step 6a-1): (Step 5a-2): (Step 6a-2): wherein, X is selected from Cl, Br or I; L is a leaving group, and the leaving group is selected from F, Br, I, R 1 is a protecting group for H or N, and the protecting group for N is selected from formyl, acetyl, trifluoroacetyl, methoxycarbonyl, ethoxycarbonyl, benzyloxycarbonyl, tert-butoxycarbonyl, allyloxycarbonyl, benzyl, p-methoxybenzyl, 2-(trimethylsilyl)ethoxymethyl or methoxymethyl; R 2 、R 3 、R 4 are independently selected from the following groups optionally substituted with R a : C 1 -C 6 alkyl, C 3 -C 6 cycloalkyl, 3- to 10-membered heterocyclic group, phenyl or 5- to 10-membered heteroaryl; Each R a is selected from F, Cl, Br, I, CN, ═O, C 1 -C 3 alkyl, OH, NH 2 , NH(C 1 -C 3 alkyl) or N(C 1 -C 3 alkyl) 2 , and when R 1 is H, the compound of formula (III) is the same as the compound of formula (IIIa), and the preparation method of the compound of formula (A) does not include step 4b.

2. The process for preparing a compound of formula (A) according to claim 1, characterized in that, L is a leaving group, and the leaving group is selected from F, Br or I.

3. The process for preparing a compound of formula (A) according to claim 1, characterized in that, the compound represented by formula (II) is selected from the compounds represented by formula (IIa): wherein, X and L are as defined in claim 1.

4. The process for preparing a compound of formula (A) according to claim 1, characterized in that, the compound represented by formula (II) is selected from the compounds represented by formula (IIb): wherein, X is as defined in claim 1.

5. The process for preparing a compound of formula (A) according to claim 1, characterized in that, the compound represented by formula (II) is selected from one of the following structures:

6. The process for preparing a compound of formula (A) according to claim 1, characterized in that, the process for preparing the compound of formula (A) comprises the following steps:

7. The process for preparing a compound of formula (A) according to claim 1, characterized in that, the process for preparing the compound of formula (A) comprises the following steps:

8. The process for preparing a compound of formula (A) according to claim 1, characterized in that, the process for preparing the compound of formula (A) comprises the following steps:

9. The process for preparing a compound of formula (A) according to claim 1, characterized in that, the process for preparing the compound of formula (II) comprises the following step 3: Among them, X, L, and R 1 As defined in claim 1, the halogenating reagent is selected from a chlorinating reagent, a brominating reagent, or an iodinating reagent.

10. The process for preparing a compound of formula (A) according to claim 9, characterized in that, the halogenating agent in step 3 is selected from N-chlorosuccinimide, N-bromosuccinimide, N-iodosuccinimide, bromine, iodine, dibromohydantoin, pyridinium tribromide, sodium bromate, 5,5-dibromo-2,2-dimethyl-4,6-dione-1,3-dioxane, 1,3-dibromo-1,3,5-triazine-2,4,6-trione, cuprous chloride or cuprous iodide.

11. The process for preparing a compound of formula (A) according to any one of claims 5-7, characterized in that, the process for preparing the compound of formula (IIc) comprises the following steps 1c, step 2c, step 3c:

12. The process for preparing a compound of formula (A) according to claim 11, characterized in that, step 1c further comprises the step of adding an acidic reagent, step 2c further comprises the step of adding a fluorine reagent and nitrite, and step 3c further comprises the step of adding a brominating reagent.

13. Compound Use as an intermediate in the preparation of a compound of formula (A) ​

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