Cyclic alpha-hydroxyimide compound and application thereof

Abstract

The invention relates to the technical field of biological medicine, and particularly discloses a cyclic alpha-hydroxyimide compound and application thereof. In the structural general formula of the compound, R1 is any one of hydrogen, C1-6 alkyl, C3-7 cycloalkyl, C3-7 heterocycloalkyl, aryl or heteroaryl substituted C1-6 alkyl; r2 is any one of hydrogen, deuterium, a C1-6 alkyl group, a C3-7 cycloalkyl group, a C3-7 heterocycloalkyl group, a C1-6 alkoxy group, a C1-6 alkylamino group, an optionally substituted aryl group or an optionally substituted heteroaryl group; x is any one of CH and N; y is any one of hydrogen, deuterium, halogen, hydroxyl, amino, cyano and a group shown in a formula (1); wherein R3 is any one of hydrogen, a hydroxyl group, an amino group, a C1-6 alkoxy group and a C1-6 alkylamino group; z is any one of CH2, NH, O and S. The compound provided by the invention has relatively strong FEN-1 inhibitory activity.

Description

Technical Field

[0001] The present application relates to the technical field of biomedicine, and more specifically, to a cyclic α-hydroxyimide compound and application thereof. Background Art

[0002] The liver is the largest digestive gland in the human body and the most active organ in the human body's metabolism. It is involved in important functions such as digestion, metabolism, immunity and nutrient storage. When the liver is diseased, many functions of the human body will malfunction. Hepatitis B (abbreviated as "HBV") is an infectious disease caused by hepatitis B virus (HBV) infection, mainly liver disease, and is a health issue of global concern. Hepatitis B patients have a high risk of dying from cirrhosis and liver cancer.

[0003] The pathogenic mechanism of HBV infection of the liver is still not very clear, but most opinions believe that: after HBV enters the human body, it will replicate and multiply in large quantities in liver cells, and damage the liver by triggering abnormal immune responses, thereby causing liver lesions. Among them, the unique replication mechanism of HBV, the vulnerability of the host immune system and other factors are involved. At present, it is generally believed that HBV is endocytosed through the sodium taurocholate cotransporter (sodium taurocholate cotransportingpolypeptide, NTCP). Inside the cell, rcDNA (double-linked relaxed circular DNA molecule) enters the cell nucleus and is repaired by host repair factors to form cccDNA (covalently closed circular DNA molecule). The formation of cccDNA is used as a viral minichromosome for HBV RNA transcription. pgRNA (pregenomic RNA, pregenomic ribonucleic acid) binds to HBV Pol (polymerase protein) in the cytosol to induce the packaging process, and then synthesizes HBV DNA. New HBV virus particles are assembled and translocated in the endoplasmic reticulum, and secreted outside the liver cells as virus particles, thereby realizing HBV replication.

[0004] The reason why HBV is difficult to eradicate and prone to relapse is that cccDNA exists stably in the nucleus of liver cells of HBV patients for a long time, and cccDNA is the starting template for HBV replication. At the same time, the host immune system cannot effectively respond to HBV, which is also a key factor in chronic HBV infection. The complete elimination of cccDNA in HBV patients is a sign of complete cure of hepatitis B.

[0005] At present, the antiviral drugs for hepatitis B are nucleotide analogues (NAs) and interferon (IFN), but neither of them can directly affect the generation process of cccDNA. NAs have a poor effect on HBV antigens, and long-term use of NAs can also cause patients to develop drug resistance. IFN also has the disadvantages of low cure rate or response rate, inconvenient administration method, and many adverse reactions and contraindications. Only by clearing cccDNA in liver cells can HBV infection be completely suppressed, thereby achieving the goal of "curing" hepatitis B.

[0006] In summary, since current antiviral drugs cannot completely eliminate HBV, there is still a huge market demand for the treatment of hepatitis B. The development of new hepatitis B drugs, especially inhibiting the generation of cccDNA through new mechanisms, will provide a new direction for the treatment of hepatitis B. Summary of the invention

[0007] The present application provides a cyclic α-hydroxyimide compound and application thereof.

[0008] Human flap endonuclease 1 (FEN-1) is a metallo-nuclease catalyzed by magnesium ions, and is mainly involved in the maturation of Okazaki fragments and the excision of 5'-flap during DNA replication. This process is substrate structure specific and only recognizes the specific 5'-flap structure of the substrate DNA, regardless of the sequence of the substrate DNA. A number of studies have shown that FEN-1 plays a vital role in the cccDNA repair process of HBV. The cyclic α-hydroxyimide compound provided in the present application has a strong FEN-1 inhibitory activity, which further inhibits the generation of hepatitis B surface antigen, hepatitis B E antigen, hepatitis B total DNA and even hepatitis B cccDNA, ultimately achieving the cure of hepatitis B.

[0009] In the first aspect, the present application provides a cyclic α-hydroxyimide compound, which adopts the following technical solution:

[0010] A cyclic α-hydroxyimide compound, the general structural formula of the cyclic α-hydroxyimide compound is as follows:

[0011]

[0012] In the general structural formula, R1 is hydrogen, C 1-6 Alkyl, C 3-7 Cycloalkyl, C 3-7 Heterocycloalkyl, aryl or heteroaryl substituted C 1-6 Any of the alkyl groups;

[0013] R2 is hydrogen, deuterium, C 1-6 Alkyl, C 3-7Cycloalkyl, C 3-7 Heterocycloalkyl, C 1-6 Alkoxy, C 1-6 any of alkylamino, optionally substituted aryl or optionally substituted heteroaryl;

[0014] X is any of CH and N;

[0015] Y is any one of hydrogen, deuterium, halogen, hydroxyl, amino, cyano, and a group as shown in formula (2);

[0016]

[0017] In the formula (2), R3 is hydrogen, hydroxyl, amino, C 1-6 Alkoxy, C 1-6 Any of alkylamino groups.

[0018] Preferably, the structural formula of the cyclic α-hydroxyimide compound is shown in formula (2) or formula (3):

[0019]

[0020] R4 is hydrogen, deuterium, C 1-6 Alkyl, C 3-7 Cycloalkyl, C 3-7 Heterocycloalkyl, C 1-6 Alkoxy, C 1-6 any of alkylamino, optionally substituted aryl or optionally substituted heteroaryl;

[0021] Z is any one of CH2, NH, O and S.

[0022] Preferably, in the general structural formula, R1 is a C 1-6 alkyl;

[0023] R2 is optionally substituted aryl or optionally substituted heteroaryl;

[0024] X is any of CH and N;

[0025] Y is a group as shown in formula (1); in formula (1), R3 is an amino group or a C 1-6 Alkoxy;

[0026] Z is CH2.

[0027] Preferably, the structural formula of the cyclic α-hydroxyimide compound is any one of formula (4) to formula (20).

[0028] Formula (4) to Formula (20) are as follows:

[0029]

[0030]

[0031] Formula (4) - 3-hydroxy-2-oxo-1H-quinoline-4-carboxylic acid ethyl ester.

[0032] Formula (5) - 1-({3-[3-(acetylamino)phenyl]phenyl}methyl)-3-hydroxy-2-oxoquinoline-4-carboxylic acid.

[0033] Formula (6)—3-hydroxy-2-oxo-1H-1,5-naphthyridine-4-carboxamide.

[0034] Formula (7)—3-Hydroxy-6-{[4-(4-hydroxyphenyl)pyridin-2-yl]methyl}-2-oxo-1H-1,5-naphthyridine-4-carboxamide.

[0035] Formula (8)—3-Hydroxy-6-{[3-(4-hydroxyphenyl)phenyl]methyl}-2-oxo-1H-1,5-naphthyridine-4-carboxamide.

[0036] Formula (9)—3-Hydroxy-6-{[5-(3-hydroxyphenyl)thiophen-2-yl]methyl}-2-oxo-1H-1,5-naphthyridine-4-carboxamide.

[0037] Formula (10)—3-hydroxy-6-{[4-(1H-pyrrolo[2,3-b]pyridin-3-yl)pyrazol-1-yl]methyl}-2-oxo-1H-1,5-naphthyridine-4-carboxamide.

[0038] Formula (11)—3-hydroxy-6-{2-[(4-hydroxyphenyl)amino]-2-oxoethyl}-2-oxo-1H-1,5-naphthyridine-4-carboxamide.

[0039] Formula (12)—3-hydroxy-2-oxo-6-{[4-(2-oxo-1H-pyridin-4-yl)pyridin-2-yl]methyl}-1H-1,5-naphthyridine-4-carboxamide.

[0040] Formula (13)—3-Hydroxy-2-oxo-6-{[5-(4-formamidophenyl)thiophen-2-yl]methyl}-1H-1,5-naphthyridine-4-carboxamide.

[0041] Formula (14)—3-Hydroxy-2-oxo-6-{[3-(4-formamidophenyl)phenyl]methyl}-1H-1,5-naphthyridine-4-carboxamide.

[0042] Formula (15)—3-Hydroxy-2-oxo-6-{[4-(5-carboxamido-1H-pyrrol-3-yl)pyrazol-1-yl]methyl}-1H-1,5-naphthyridine-4-carboxamide.

[0043] Formula (16)—3-Hydroxy-2-oxo-6-{[6-(1H-pyrrolo[2,3-b]pyridin-3-yl)pyridin-3-yl]methyl-1H-1,5-naphthyridine-4-carboxamide.

[0044] Formula (17)—3-Hydroxy-2-oxo-6-{[5-(1H-pyrrolo[2,3-b]pyridin-3-yl)thiophen-2-yl]methyl}-1H-1,5-naphthyridine-4-carboxamide.

[0045] Formula (18)—3-Hydroxy-2-oxo-6-({5-[3-(4-methylpiperazin-1-yl)phenyl]thiophen-2-yl}methyl)-1H-1,5-naphthyridine-4-carboxamide.

[0046] Formula (19)—4-{3-[(8-Formamido-7-hydroxy-6-oxo-5H-1,5-naphthyridin-2-yl)methyl]phenyl}benzoic acid.

[0047] Formula (20)—3-Hydroxy-2-oxo-6-({3-[(4-hydroxyphenyl)methyl]phenyl}methyl)-1H-1,5-naphthyridine-4-carboxamide.

[0048] In addition, the cyclic α-hydroxyimide compound can also be any one of 3-hydroxy-2-oxo-1H-quinoline-4-carboxylic acid, 6-{[4-(3-chloro-4-hydroxyphenyl)piperazine-1-yl]methyl}-3-hydroxy-2-oxo-1H-1,5-naphthyridine-4-carboxamide, and 3-hydroxy-6-[2-(5-hydroxybenzo[d]thiazol-2-yl)ethyl]-2-oxo-1H-1,5-naphthyridine-4-carboxamide.

[0049] In summary, this application has the following beneficial effects:

[0050] The cyclic α-hydroxyimide compound provided in the present application has strong FEN-1 inhibitory activity and can inhibit the production of hepatitis B surface antigen, hepatitis B E antigen, hepatitis B total DNA and cccDNA at the cellular level, and is expected to be developed into a new anti-hepatitis B drug. BRIEF DESCRIPTION OF THE DRAWINGS

[0051] Figure 1 It is a synthetic route diagram of a specific compound represented by formula (3).

[0052] Figure 2 It is a synthetic route diagram of a specific compound represented by formula (2).

[0053] Figure 3 This is the inhibition rate of the compound provided in Example 13 of the present application on HbeAg.

[0054] Figure 4 The inhibitory effect of the compound provided in Example 13 of the present application on cccDNA (Southern-blot results). DETAILED DESCRIPTION

[0055] Explanation of terms

[0056] In order to describe the content of this application more clearly, the terms involved in this application are explained as follows. Those skilled in the art can understand the following terms in combination with relevant technologies.

[0057] The term "C 1-6 The term “alkyl” alone or in combination means a saturated straight-chain or branched alkyl group containing 1 to 6, especially 1 to 4 carbon atoms, including methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, 2-pentyl, 3-pentyl, 2-methyl-2-butyl, 3-methyl-2-butyl, 3-methyl-1-butyl, 2-methyl-1-butyl, n-hexyl, 2-hexyl, 3-hexyl, 2-methyl-2-pentyl, 3-methyl-2-pentyl, 4-methyl-2-pentyl, 3-methyl-3-pentyl, 2-methyl-3-pentyl, 2,3-dimethyl-2-butyl, 3,3-dimethyl-2-butyl, etc.

[0058] Preferably, "C 1-6 The "alkyl" is any one of methyl, ethyl, n-propyl, isopropyl and tert-butyl.

[0059] The term "C 3-7 "Cycloalkyl" alone or in combination means a saturated cycloalkyl group containing 3-7, especially 3-6 carbon atoms, including cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and the like.

[0060] Preferably, "C 3-7 The "cycloalkyl" is any of cyclopropyl, cyclopentyl and cyclohexyl.

[0061] The term "C 3-7 The term "heterocycloalkyl" refers to a cycloalkyl group containing 3 to 7 carbon atoms and heteroatoms in a monocyclic heterocycloalkyl group, including aziridine, azetidinyl, oxetanyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, piperidinyl, morpholinyl, piperazinyl, etc.

[0062] The term "C 1-6 "Alkoxy" alone or in combination means a group C 1-6 Alkyl-O-, where "C1-6 "Alkyl" is as defined above. "C 1-6 “Alkoxy” includes, but is not limited to, methoxy (-OCH3), ethoxy (-OCH2CH3), n-propoxy (-OCH2CH2CH3), isopropoxy (-OCH(CH3)2), n-butoxy (-OCH2CH2CH2CH3), sec-butoxy (-OCH(CH3)CH2CH3), isobutoxy (-OCH2CH(CH3)2), tert-butoxy (-OC(CH3)3), n-pentoxy (-OCH2CH2CH2CH2CH3), neopentoxy (-OCH2C(CH3)3), and the like.

[0063] The term "C 1-6 "Alkylamino" alone or in combination means a group C 1-6 Alkyl-N-, where "C 1-6 "Alkyl" is as defined above. "C 1-6 The “alkylamino” includes methylamino, ethylamino, propylamino, isopropylamino, n-butylamino, isobutylamino, 2-butylamino, tert-butylamino, n-pentylamino, 2-pentylamino, 3-pentylamino, 2-methyl-2-butylamino, 3-methyl-2-butylamino, 3-methyl-1-butylamino, 2-methyl-1-butylamino, n-hexylamino, 2-hexylamino, 3-hexylamino, 2-methyl-2-pentylamino, 3-methyl-2-pentylamino, 4-methyl-2-pentylamino, 3-methyl-3-pentylamino, 2-methyl-3-pentylamino, 2,3-dimethyl-2-butylamino, 3,3-dimethyl-2-butylamino and the like.

[0064] Preferably, "C 1-6 The "alkylamino" is any one of methylamino, ethylamino, isopropylamino and tert-butylamino.

[0065] The term "heteroaryl" alone or in combination means a group having one or more rings of 5 to 20 atoms, wherein at least one atom in one ring is a heteroatom selected from nitrogen, oxygen, sulfur; and wherein at least one ring containing heteroatoms is aromatic. In heteroaryl groups comprising 2 or more fused rings, the ring with non-heteroatoms may be a carbocyclic ring. Heteroaryl groups may be substituted.

[0066] The term "pharmaceutically acceptable salt" means that the compounds of the present application exist in the form of their pharmaceutical salts, including acid addition salts and base addition salts. "Pharmaceutically acceptable salts" are described in pharmaceutically salts described by SM Berge in J. Pharmaceutical Sciences (Volume 66: 1-19, 1977). In the present application, "pharmaceutically acceptable, non-toxic acid addition salts" means salts formed by the compounds in the present application with organic acids or inorganic acids, and organic acids or inorganic acids include but are not limited to hydrochloric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, nitric acid, peramic acid, acetic acid, oxalic acid, maleic acid, fumaric acid, tartaric acid, benzenesulfonic acid, methanesulfonic acid, salicylic acid, succinic acid, citric acid, lactic acid, propionic acid, benzoic acid, p-toluenesulfonic acid, malic acid, etc. "Pharmaceutically acceptable, non-toxic base addition salt" refers to a salt formed by a compound of the present invention with an organic base or an inorganic base, including but not limited to alkali metal salts, such as carbendazim, sodium or potassium salts; alkaline earth metal salts, such as calcium or magnesium salts; organic base salts, such as salts formed by addition of an organic base containing an N group or N + (C 1-6 preferably, lithium hydroxide, sodium hydroxide, potassium hydroxide, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium bicarbonate, magnesium carbonate, calcium carbonate, ammonia water, triethylamine, tetrabutylammonium hydroxide, etc. "Pharmaceutically acceptable salts" can be synthesized by general chemical methods.

[0067] The term "hydrate" refers to an association formed between water and a compound in the present application.

[0068] The term "solvate" refers to an association formed by one or more solvent molecules and the compound in the present application. Solvents that form solvates include, but are not limited to, water, methanol, ethanol, isopropanol, ethyl acetate, tetrahydrofuran, N,N-dimethylformamide, dimethyl sulfoxide, and the like.

[0069] The term "isomer" includes all isomeric forms, including enantiomers, diastereomers, tautomers and geometric isomers (including cis-trans isomers). Therefore, the single stereochemical isomers of the compounds involved in the present application or mixtures of their enantiomers, diastereomers, tautomers or geometric isomers (cis-trans isomers) all fall within the scope of protection claimed in the present application.

[0070] The term "prodrug" refers to a chemical derivative of the compound of the present application, which is converted into a compound represented by formula (1) through a chemical reaction in vivo.

[0071] The term "isotopic derivative" refers to an isotopic derivative obtained by replacing hydrogen atoms in the compound represented by formula (1) with 1 to 6 deuterium atoms, or an isotopic derivative obtained by replacing carbon atoms in the compound represented by formula (1) with 1 to 3 carbon 14 atoms.

[0072] The present application provides a cyclic α-hydroxyimide compound, and the general structural formula of the cyclic α-hydroxyimide compound is as follows:

[0073]

[0074] In the general structural formula, R1 is hydrogen, C 1-6 Alkyl, C 3-7 Cycloalkyl, C 3-7 Heterocycloalkyl, aryl or heteroaryl substituted C 1-6 Any of the alkyl groups;

[0075] R2 is hydrogen, deuterium, C 1-6 Alkyl, C 3-7 Cycloalkyl, C 3-7 Heterocycloalkyl, C 1-6 Alkoxy, C 1-6 any of alkylamino, optionally substituted aryl or optionally substituted heteroaryl;

[0076] X is any of CH and N;

[0077] Y is any one of hydrogen, deuterium, halogen, hydroxyl, amino, cyano, and a group as shown in formula (2);

[0078]

[0079] In the formula (1), R3 is hydrogen, hydroxyl, amino, C 1-6 Alkoxy, C 1-6 Any of alkylamino groups.

[0080] Preferably, the structural formula of the cyclic α-hydroxyimide compound is shown in formula (2) or formula (3):

[0081]

[0082] R4 is hydrogen, deuterium, C 1-6 Alkyl, C 3-7 Cycloalkyl, C 3-7 Heterocycloalkyl, C 1-6 Alkoxy, C 1-6 any of alkylamino, optionally substituted aryl or optionally substituted heteroaryl;

[0083] Z is any one of CH2, NH, O and S.

[0084] The present application also provides a method for preparing the above-mentioned cyclic α-hydroxyimide compound. The following describes the synthesis route of the cyclic α-hydroxyimide compound provided by the present application to further describe the technical solution of the present application.

[0085] like Figure 1 The product 8 in the synthetic route shown is a specific compound of the above formula (3).

[0086] When X in formula (3) is N, Y is CONH2, and Z is CH2, formula (3) is Figure 1 The product 8 in the synthetic route shown in .

[0087] refer to Figure 1 The synthetic route of product 8 specifically comprises the following steps:

[0088] (1) Compound 1 reacts with tert-butyl cyanoacetate to obtain product 2;

[0089] (2) Product 2 is deacylated under acidic conditions to obtain product 3;

[0090] (3) Reducing the nitro group of product 3 to obtain product 4;

[0091] (4) product 4 reacts with ethyl oxalyl chloride to obtain product 5;

[0092] (5) Product 5 is cyclized under alkaline conditions to obtain product 6;

[0093] (6) Product 6 is reacted with substituted ethylboronic acid or substituted Boc-methane to obtain product 7;

[0094] (7) Product 7 is hydrolyzed with polyphosphoric acid (PPA) to obtain product 8.

[0095] like Figure 2 The product 18 in the synthetic route shown is a specific compound of the above formula (2).

[0096] When X in formula (2) is CH and Y is the structure shown in formula (1), formula (2) is Figure 2 The product 12 in the synthetic route shown in .

[0097] refer to Figure 2 The synthetic route of product 12 specifically includes the following steps:

[0098] (1) Compound 9 reacts with trimethylsilylated diazomethane to obtain product 10;

[0099] (2) Product 10 reacts with bromide to obtain product 11;

[0100] (3) Product 11 is demethylated in BBr3 to give product 12.

[0101] The abbreviations of materials commonly involved in this application are shown in Table 1.

[0102] Table 1 Abbreviations of materials commonly involved in this application

[0103]

[0104] The present application is further described by the following examples, however, these examples should not be construed as limiting the scope of the present application.

[0105] Example

[0106] The following describes the general experimental conditions in the examples of this application.

[0107] First, the reactions in the examples of the present application were all carried out under nitrogen protection.

[0108] Furthermore, the intermediates and final products in the examples of the present application were separated and purified by thin layer chromatography silica gel plates (Qingdao Bangkai), rapid preparative liquid phase system (Biotage Isolera Flash) and preparative high performance liquid chromatography system (Agilent 1290 Infinity II LC System, with Welch Xtimate C18 reverse phase chromatography column).

[0109] Furthermore, the liquid chromatography-mass spectrometry (LC-MS) instrument uses Waters ACQUITY Arc equipped with QDaDetector. The mass spectrometry (MS) uses an ESI source and only indicates the molecular weight M of the parent molecule, usually reporting [M+H] + . The injection volume is determined by the sample concentration: the flow rate is mL / min. The peak of the high performance liquid chromatography (HPLC) is recorded and read by UV Vis wavelengths at 220nm and 254nm. The mobile phases are A and B, and the gradient elution conditions are shown in Table 2 below. The elution conditions mentioned in Table 2 can achieve the elution of the α-hydroxy ketone nitrogen-containing fused ring compounds in the following examples.

[0110] Table 2 Gradient elution conditions

[0111]

[0112] Further, the nuclear magnetic resonance (NMR) spectra are obtained using a 400 MHz nuclear magnetic resonance spectrometer, often using CDCl3 and DMSO-d6 as solvents, and reporting chemical shifts (δ) in ppm (parts per million). The descriptions of the various peaks are as follows: s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), dd (doublet of doublets). Coupling constants are expressed in Hz (Hertz).

[0113] The names, structural formulas and preparation processes of the cyclic α-hydroxyimide compounds provided in Examples 1-9 are specifically shown in Tables 3 and 4.

[0114] Table 3 Names and structural formulas of intermediates in each step

[0115]

[0116]

[0117]

[0118]

[0119] Table 4 Name, structural formula and preparation process of cyclic α-hydroxyimide compounds provided in the examples (I)

[0120]

[0121]

[0122]

[0123] Example 1

[0124] This embodiment provides 3-hydroxy-2-oxo-1H-quinoline-4-carboxylic acid ethyl ester, the structural formula of which is shown in formula (4), and the specific preparation method is as follows.

[0125]

[0126] Indigo carmine (4.42 g, 10 mmol) was dissolved in 48 mL EtOH / DMF (volume ratio of 5:1), 1,8-diazabicycloundec-7-ene (DBU) (0.7 mL, 1.5 mmol) was quickly added, and the mixture was stirred evenly at room temperature. Then, ethyl diazoacetate (7.5 mL, 20 mmol) was added under nitrogen protection, and the mixture was reacted at room temperature for 15 h. The reaction solution was concentrated under reduced pressure to remove most of the EtOH, and 35 mL of 1M dilute hydrochloric acid was slowly added and stirred for 2 h. The reaction solution was filtered, the filter cake was collected, and washed with 40 mL of water and 40 mL of EtOH, respectively. It was purified by reverse phase high performance liquid chromatography (Pre-HPLC) and dried in a vacuum oven to obtain a white solid - 3-hydroxy-2-oxo-1H-quinoline-4-carboxylic acid ethyl ester (yield 42 g, 46%).

[0127] The test results are as follows:

[0128] MS (ESI) m / z = 234.1 [M + H] + .

[0129] 1 H NMR (DMSO-d6, 400MHz): δ 12.30 (s, 1H), 10.22 (s, 1H), 7.43-7.29 (m, 3H), 7.24-7.12 (m, 1H), 4.40 (q, J=7.2Hz, 2H), 1.33 (t, J=7.2Hz, 3H).

[0130] Example 2

[0131] This embodiment provides 1-({3-[3-(acetylamino)phenyl]phenyl}methyl)-3-hydroxy-2-oxoquinoline-4-carboxylic acid, whose structural formula is shown in formula (5). The specific preparation method is as follows, see Table 3 and Table 4 for details.

[0132] (1) Preparation of ethyl 3-methoxy-2-oxo-1H-quinoline-4-carboxylate (2A), the structural formula of which is shown in formula (21).

[0133] 3-Hydroxy-2-oxo-1H-quinoline-4-carboxylic acid ethyl ester (the product prepared in Example 1) was dissolved in THF / MeOH; TMSCHN2 (trimethylsilyl)diazomethane) was then added to the solution and stirred at room temperature for 1 hour. After the reaction was completed, the mixture was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to obtain a white solid, 3-methoxy-2-oxo-1H-quinoline-4-carboxylic acid ethyl ester (2A).

[0134] (2) Preparation of ethyl 1-(3-bromobenzyl)-3-methoxy-2-oxoquinoline-4-carboxylate (2B), the structural formula of which is shown in formula (22).

[0135] 3-Methoxy-2-oxo-1H-quinoline-4-carboxylic acid ethyl ester (2A) was dissolved in 5 mL of DMF, sodium hydride was slowly added in an ice-water bath, stirring was continued for 20 min, and then 3-bromobenzyl bromide was added, and the reaction was continued for 1 h. After the reaction was completed, 50 mL of water was added and extracted with EA (20 mL×3), the organic phase was washed with saturated brine, and then the organic phase was concentrated, and the crude product was purified by silica gel column chromatography to obtain a white solid - 1-(3-bromobenzyl)-3-methoxy-2-oxoquinoline-4-carboxylic acid ethyl ester (2B).

[0136] (3) Preparation of ethyl 1-({3-[3-(acetylamino)phenyl]phenyl}methyl)-3-methoxy-2-oxoquinoline-4-carboxylate (2C), the structural formula of which is shown in formula (23).

[0137] 1-(3-Bromobenzyl)-3-methoxy-2-oxoquinoline-4-carboxylic acid ethyl ester (2B) was dissolved in 1,4-dioxane, 3-acetamidophenylboronic acid pinacol ester and cesium carbonate were added to the solution, and Pd(dppf)Cl2 (catalyst) was added after stirring and mixing, and then the temperature was raised to 100°C for 16 hours. After the reaction was completed, the mixture was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography to obtain a colorless oily substance, 1-({3-[3-(acetylamino)phenyl]phenyl}methyl)-3-methoxy-2-oxoquinoline-4-carboxylic acid ethyl ester (2C).

[0138] (4) Preparation of ethyl 1-({3-[3-(acetylamino)phenyl]phenyl}methyl)-3-hydroxy-2-oxoquinoline-4-carboxylate (2D), the structural formula of which is shown in formula (24).

[0139] 1-({3-[3-(acetylamino)phenyl]phenyl}methyl)-3-methoxy-2-oxoquinoline-4-carboxylic acid ethyl ester (2C) was dissolved in DCM, 1M BBr3 solution was added to the solution, and then stirred at room temperature for 10 minutes. After the reaction was completed, 40 mL of water was added to quench, and then DCM was added for extraction (10 mL×3), and the organic phase was collected, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product of 1-({3-[3-(acetylamino)phenyl]phenyl}methyl)-3-hydroxy-2-oxoquinoline-4-carboxylic acid ethyl ester (2D).

[0140] (5) Prepare 1-({3-[3-(acetylamino)phenyl]phenyl}methyl)-3-hydroxy-2-oxoquinoline-4-carboxylic acid, the structural formula of which is shown in formula (5).

[0141] Ethyl 1-({3-[3-(acetylamino)phenyl]phenyl}methyl)-3-hydroxy-2-oxoquinoline-4-carboxylate (2D) was dissolved in THF, and an aqueous solution of LiOH was added to the solution at room temperature, and then reacted at room temperature for 16 hours. After the reaction was completed, the solution was concentrated under reduced pressure and purified by Pre-HPLC to obtain a white solid, 1-({3-[3-(acetylamino)phenyl]phenyl}methyl)-3-hydroxy-2-oxoquinoline-4-carboxylic acid.

[0142] Example 3

[0143] This embodiment provides 3-hydroxy-2-oxo-1H-1,5-naphthyridine-4-carboxamide, the structural formula of which is shown in formula (6), and the specific preparation method is as follows, as shown in Table 3 and Table 4.

[0144] (1) Preparation of tert-butyl (2-bromopyridin-3-yl)carbamate (3A), the structural formula of which is shown in formula (25).

[0145] 2-Bromonicotinic acid was dissolved in tert-butyl alcohol, triethylamine and diphenylphosphoryl azide were added, and the temperature was raised to 80°C for reaction for 3 hours. The crude product was concentrated to obtain a crude product, which was purified by column chromatography to obtain a white solid - tert-butyl (2-bromopyridin-3-yl)carbamate (3A).

[0146] (2) Preparation of tert-butyl (2-formylpyridin-3-yl)carbamate (3B), the structural formula of which is shown in formula (26).

[0147] Dissolve tert-butyl (2-bromopyridin-3-yl)carbamate (3A) in THF, cool to -68°C under nitrogen protection, add n-butyl lithium dropwise, stir at the temperature for 10 min, add DMF, and keep the temperature for 2 h. After the reaction, add 200 mL of saturated brine and 600 mL of EA, separate the liquids, dry and concentrate the organic phase, and purify the crude product by column chromatography to obtain a light yellow oily substance - tert-butyl (2-formylpyridin-3-yl)carbamate (3B).

[0148] (3) Prepare 3-aminopyridinecarboxaldehyde (3C), whose structural formula is shown in formula (27).

[0149] Dissolve tert-butyl (2-formylpyridin-3-yl)carbamate (3B) in DCM, add TFA, react at 30°C for 4 hours, and concentrate after the reaction is complete. Dilute with DCM, add saturated sodium bicarbonate solution to adjust pH to 7-8, separate and collect the organic phase, dry the organic phase over anhydrous sodium sulfate, and concentrate to obtain a yellow solid - 3-aminopyridinecarboxaldehyde (3C).

[0150] (4) Prepare 3-[(4-methoxybenzyl)oxy]-1H-1,5-naphthyridin-2-one (3D), whose structural formula is shown in formula (28).

[0151] 3-Aminopyridinecarboxaldehyde (3C) was dissolved in THF, ethyl 2-[(4-methoxybenzyl)oxy]acetate was added, and LiHMDS (lithium hexamethyldisilazide) was added dropwise at -10°C. The mixture was reacted at room temperature for 16 hours. After the reaction was completed, a yellow solid was obtained by filtration, namely 3-[(4-methoxybenzyl)oxy]-1H-1,5-naphthyridine-2-one (3D).

[0152] (5) Prepare 3-hydroxy-1H-1,5-naphthyridin-2-one (3E), whose structural formula is shown in formula (29).

[0153] 3-[(4-methoxybenzyl)oxy]-1H-1,5-naphthyridine-2-one (3D) was dissolved in DCM, TFA was added, and the mixture was reacted at room temperature for 16 h. After the reaction was completed, the mixture was concentrated and 200 mL of ether was added to slurry to obtain a brown solid, 3-hydroxy-1H-1,5-naphthyridine-2-one (3E).

[0154] (6) Prepare 4-bromo-3-hydroxy-1H-1,5-naphthyridin-2-one (3F), the structural formula of which is shown in formula (30).

[0155] 3-Hydroxy-1H-1,5-naphthyridin-2-one (3E) was dissolved in acetic acid, and NBS was added to react at room temperature for 2 hours. After the reaction was completed, a yellow solid, 4-bromo-3-hydroxy-1H-1,5-naphthyridin-2-one (3F), was obtained by filtration.

[0156] (7) Prepare 4-bromo-1-(4-methoxybenzyl)-3-[(4-methylbenzyl)oxy]-1H-1,5-naphthyridin-2-one (3G), the structural formula of which is shown in formula (31).

[0157] 4-Bromo-3-hydroxy-1H-1,5-naphthyridine-2-one (3F) was dissolved in DMF, and 4-methoxybenzyl chloride, cesium carbonate, and potassium iodide were added, and the mixture was reacted at room temperature for 3 hours. After the reaction was completed, 100 mL of water was added for dilution, and the mixture was extracted with EA (100 mL × 3). The organic phases were combined, washed with saturated brine, dried, and concentrated. The crude product was purified by column chromatography to obtain a white solid, 4-bromo-1-(4-methoxybenzyl)-3-[(4-methylbenzyl)oxy]-1H-1,5-naphthyridine-2-one (3G).

[0158] (8) Prepare 3-[(4-methoxybenzyl)oxy]-2-oxo-1,2-dihydro-1,5-naphthyridine-4-carbonitrile (3H), the structural formula of which is shown in formula (32).

[0159] 4-Bromo-1-(4-methoxybenzyl)-3-[(4-methylbenzyl)oxy]-1H-1,5-naphthyridin-2-one (3G) was dissolved in dimethylacetamide (DMA), and zinc cyanide, zinc powder, 1,1-bis(diphenylphosphino)ferrocene, and Pd2(Dba)3 were added, and the mixture was reacted at 135°C for 16 hours. After the reaction was completed, the mixture was concentrated and purified by reverse phase column chromatography to obtain a white solid, 3-[(4-methoxybenzyl)oxy]-2-oxo-1,2-dihydro-1,5-naphthyridin-4-carbonitrile (3H).

[0160] (9) Prepare 3-hydroxy-2-oxo-1H-1,5-naphthyridine-4-carboxamide, the structural formula of which is shown in formula (6).

[0161] 3-[(4-methoxybenzyl)oxy]-2-oxo-1,2-dihydro-1,5-naphthyridine-4-carbonitrile (3H) was dissolved in concentrated sulfuric acid and heated to 60°C for 2 hours. After the reaction was completed, the mixture was filtered and the filter cake was slurried with 3 mL of MeOH to obtain a black solid, 3-hydroxy-2-oxo-1H-1,5-naphthyridine-4-carboxamide.

[0162] Example 4

[0163] This embodiment provides 3-hydroxy-6-{[4-(4-hydroxyphenyl)pyridin-2-yl]methyl}-2-oxo-1H-1,5-naphthyridine-4-carboxamide, whose structural formula is shown in formula (7). The specific preparation method is as follows, see Table 3 and Table 4 for details.

[0164] (1) Preparation of tert-butyl 2-(6-chloro-3-nitropyridin-2-yl)-2-cyanoacetate (4A), the structural formula of which is shown in formula (33).

[0165] 2,6-Dichloro-3-nitropyridine was dissolved in THF, and then anhydrous potassium carbonate and tert-butyl cyanoacetate were added in sequence, stirred evenly, and heated to 70°C for 12 hours. After the reaction was completed, the filtrate was filtered and concentrated to obtain a brown oily substance, tert-butyl 2-(6-chloro-3-nitropyridine-2-yl)-2-cyanoacetate (4A).

[0166] (2) Preparation of 2-(6-chloro-3-nitropyridin-2-yl)acetonitrile (4B), the structural formula of which is shown in formula (34).

[0167] Slowly add 6M hydrochloric acid solution to the MeCN solution of tert-butyl 2-(6-chloro-3-nitropyridin-2-yl)-2-cyanoacetate (4A), stir evenly and heat to 70°C for 6 hours. After the reaction, add saturated sodium carbonate solution to adjust the pH to 8, then extract with EA (300mL×2), wash the organic phase with saturated brine 3 times, dry the organic phase with anhydrous sodium sulfate and concentrate to obtain a brown oily substance - 2-(6-chloro-3-nitropyridin-2-yl)acetonitrile (4B).

[0168] (3) Preparation of 2-(6-chloro-3-aminopyridin-2-yl)acetonitrile (4C), the structural formula of which is shown in formula (35).

[0169] 2-(6-chloro-3-nitropyridin-2-yl)acetonitrile (4B) and ammonium chloride were added to a 1L round-bottom flask, MeOH and water were added to dissolve, zinc flakes were slowly added in an ice-water bath, the ice-water bath was removed after the addition, and the reaction was continued for 2 hours. After the reaction was completed, a saturated sodium carbonate solution was added to adjust the pH to 8, and then EA was extracted (300mL×2), the organic phase was washed 3 times with saturated brine, the organic phase was dried over anhydrous sodium sulfate and concentrated, and the crude product was purified by silica gel column chromatography to obtain a yellow solid - 2-(6-chloro-3-aminopyridin-2-yl)acetonitrile (4C).

[0170] (4) Preparation of ethyl {[6-chloro-2-(cyanomethyl)pyridin-3-yl]carbamoyl}carboxylate (4D), the structural formula of which is shown in formula (36).

[0171] 2-(6-chloro-3-aminopyridin-2-yl)acetonitrile (4D) and pyridine were added to a 500 mL round-bottom flask, DCM was added to dissolve, and ethyl oxalyl chloride was slowly added under an ice-water bath. After the addition, the ice-water bath was removed and the reaction was continued for 1 hour. After the reaction was completed, the reaction solution was concentrated, and the crude product was purified by silica gel column chromatography to obtain a yellow solid, ethyl {[6-chloro-2-(cyanomethyl)pyridin-3-yl]carbamoyl}carboxylate (4D).

[0172] (5) Prepare 6-chloro-3-hydroxy-2-oxo-1,2-dihydro-1,5-naphthyridine-4-carbonitrile (4E), the structural formula of which is shown in formula (37).

[0173] Ethyl {[6-chloro-2-(cyanomethyl)pyridin-3-yl]carbamoyl}carboxylate (4D) was dissolved in THF, and t-BuOK (potassium tert-butoxide) was added under an ice-water bath. After the addition, the ice-water bath was removed and the reaction was continued for 2 hours. After the reaction was completed, the reaction solution was concentrated and filtered, and the filter cake was washed with water and EtOH respectively to obtain a white solid - 6-chloro-3-hydroxy-2-oxo-1,2-dihydro-1,5-naphthyridine-4-carbonitrile (4E).

[0174] (6) Preparation of tert-butyl 2-(4-bromopyridin-2-yl)acetate (4F), the structural formula of which is shown in formula (38).

[0175] Dissolve 4-bromo-2-methylpyridine in THF, cool to -70°C, slowly add lithium diisopropylamide under nitrogen protection, and stir for 1.5h after addition. Then slowly add Boc anhydride THF solution to the above solution, slowly warm to room temperature after addition and react overnight. After the reaction is completed, add saturated ammonium chloride solution to quench, then extract with EA (100mL×2), wash the organic phase with saturated brine 3 times, dry the organic phase with anhydrous sodium sulfate and concentrate, and purify the crude product by silica gel column chromatography to obtain a yellow oily substance - tert-butyl 2-(4-bromopyridin-2-yl)acetate (4F).

[0176] (7) Preparation of tert-butyl 2-(4-(4-methoxyphenyl)pyridin-2-yl)acetate (4G), the structural formula of which is shown in formula (39).

[0177] Add tert-butyl 2-(4-bromopyridin-2-yl)acetate (4F), 4-methoxyphenylboronic acid, potassium carbonate to a 100 mL round-bottom flask, add 1,4-dioxane and water respectively, and stir to dissolve. Then add [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride to the solution, heat to 85°C under nitrogen protection, and react for 2 hours. After the reaction is completed, filter and collect the filtrate and concentrate it. The crude product is purified by silica gel column chromatography to obtain a yellow oily substance - tert-butyl 2-(4-(4-methoxyphenyl)pyridin-2-yl)acetate (4G).

[0178] (8) Prepare 3-hydroxy-6-{[4-(4-methoxyphenyl)pyridin-2-yl]methyl}-2-oxo-1,2-dihydro-1,5-naphthyridine-4-carbonitrile (4H), the structural formula of which is shown in formula (40).

[0179] To a 50 mL round-bottom flask, tert-butyl 2-(4-(4-methoxyphenyl)pyridin-2-yl)acetate (4G), 6-chloro-3-hydroxy-2-oxo-1,2-dihydro-1,5-naphthyridine-4-carbonitrile (4E), lithium tert-butoxide, [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium, and imid-phos ligand were added, 1,4-dioxane was added to dissolve, and the temperature was raised to 130°C under nitrogen protection and reacted for 16 hours. After the reaction was completed, the reaction was concentrated, and the crude product was purified by Prep-HPLC to obtain a brown-yellow solid - 3-hydroxy-6-{[4-(4-methoxyphenyl)pyridin-2-yl]methyl}-2-oxo-1,2-dihydro-1,5-naphthyridine-4-carbonitrile (4H).

[0180] (9) Prepare 3-hydroxy-6-{[4-(4-methoxyphenyl)pyridin-2-yl]methyl}-2-oxo-1,2-dihydro-1,5-naphthyridine-4-carboxamide (4I), the structural formula of which is shown in formula (41).

[0181] 3-Hydroxy-6-{[4-(4-methoxyphenyl)pyridin-2-yl]methyl}-2-oxo-1,2-dihydro-1,5-naphthyridine-4-carbonitrile (4H) was added to a 50 mL round flask, and PPA was added at the same time. The temperature was raised to 110°C and stirred for 1 hour. After the reaction was completed, water was added to quench the reaction and the mixture was filtered. The filter cake was washed with EtOH to obtain a yellow solid, 3-hydroxy-6-{[4-(4-methoxyphenyl)pyridin-2-yl]methyl}-2-oxo-1,2-dihydro-1,5-naphthyridine-4-carboxamide (4I).

[0182] (10) Prepare 3-hydroxy-6-{[4-(4-hydroxyphenyl)pyridin-2-yl]methyl}-2-oxo-1H-1,5-naphthyridine-4-carboxamide, the structural formula of which is shown in formula (7).

[0183] 3-Hydroxy-6-{[4-(4-methoxyphenyl)pyridin-2-yl]methyl}-2-oxo-1,2-dihydro-1,5-naphthyridine-4-carboxamide (4I) was added to a 50 mL round flask, and DCM was added to dissolve it. Then, a 2M solution of BBr3 in DCM was added under an ice-water bath. After the addition, the ice-water bath was removed and the reaction was allowed to proceed for 1 hour. After the reaction was completed, the mixture was concentrated, and the crude product was purified by Prep-HPLC to obtain a light yellow solid, 3-hydroxy-6-{[4-(4-hydroxyphenyl)pyridin-2-yl]methyl}-2-oxo-1H-1,5-naphthyridine-4-carboxamide.

[0184] Example 5

[0185] This embodiment provides 3-hydroxy-6-{[3-(4-hydroxyphenyl)phenyl]methyl}-2-oxo-1H-1,5-naphthyridine-4-carboxamide. Its structural formula is shown in formula (8). The specific preparation method is as follows, as shown in Table 3.

[0186]

[0187] (1) Preparation of tert-butyl 2-(3-bromophenyl)acetate (5A), the structural formula of which is shown in formula (42).

[0188] 2-(3-bromophenyl)acetic acid (5 g, 23.25 mmol) and DMAP (1.42 g, 11.63 mmol) were added to a 250 mL round-bottom flask, and tert-butyl alcohol (50 mL) and THF (50 mL) were added respectively, and stirred to dissolve. Then Boc anhydride (10.15 g, 46.50 mmol) was added to the solution, and the reaction was carried out at 30° C. for 12 h. After the reaction was completed, the reaction solution was concentrated, and the crude product was purified by silica gel column chromatography to obtain a colorless oil - tert-butyl 2-(3-bromophenyl)acetate (5A) (yield 6.3 g, yield 99.93%).

[0189] (2) Preparation of tert-butyl 2-(4'-methoxy-[1,1'-biphenyl]-3-yl)acetate (5B), the structural formula of which is shown in formula (43).

[0190] To a 100 mL round-bottom flask, tert-butyl 2-(3-bromophenyl)acetate (5A) (3.30 g, 12.17 mmol), 4-methoxyphenylboronic acid (2.22 g, 14.60 mmol) and potassium carbonate (5.05 g, 36.51 mmol) were added in sequence, and 1,4-dioxane (55 mL) and water (11 mL) were added respectively, and stirred to dissolve. Then, [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (994 mg, 1.22 mmol) was added to the solution, and the temperature was raised to 90°C under nitrogen protection, and the reaction was carried out for 12 h. After the reaction was completed, the filtrate was collected by filtration, extracted with EA (20 mL×3), and the organic phase was washed with 10 mL of saturated brine, dried over anhydrous sodium sulfate and concentrated. The crude product was purified by silica gel column chromatography to obtain a colorless oily substance, tert-butyl 2-(4'-methoxy-[1,1'-biphenyl]-3-yl)acetate (5B) (yield 1.6 g, yield rate 44.06%).

[0191] Then, using tert-butyl 2-(4'-methoxy-[1,1'-biphenyl]-3-yl)acetate (5B) (200 mg, 0.902 mmol) as raw material, a yellow solid, 3-hydroxy-6-{[3-(4-hydroxyphenyl)phenyl]methyl}-2-oxo-1H-1,5-naphthyridine-4-carboxamide, was prepared in a manner similar to steps (8)-(10) of Example 4 (yield 23 mg, yield rate 52.50%).

[0192] The test results are as follows:

[0193] MS (ESI) m / z = 388.2 [M + H] + .

[0194] 1 H NMR (DMSO-d6, 400MHz): δ16.21 (s, 1H), 12.25-11.97 (m, 1H), 10.78 (s, 1H), 10.08-9.88 (m , 1H), 9.65-9.54(m, 1H), 8.73(s, 1H), 7.79-7.34(m, 5H), 7.25-6.82(m, 4H), 4.26(d, 2H).

[0195] Example 6

[0196] This embodiment provides 3-hydroxy-6-{[5-(3-hydroxyphenyl)thiophen-2-yl]methyl}-2-oxo-1H-1,5-naphthyridine-4-carboxamide. Its structural formula is shown in formula (9). The specific preparation method is as follows, as shown in Table 3.

[0197]

[0198] (1) Preparation of tert-butyl 2-(5-bromothiophen-2-yl)acetate (6A), the structural formula of which is shown in formula (44).

[0199] 5-Bromo-2-thiopheneacetic acid (2 g, 9.05 mmol) and DMAP (533 mg, 4.52 mmol) were added to a 250 mL round-bottom flask, and tert-butyl alcohol (20 mL) and THF (20 mL) were added respectively, and stirred to dissolve. Then Boc anhydride (3.95 g, 18.09 mmol) was added to the solution, and the reaction was carried out at 30 ° C for 12 h. After the reaction was completed, the reaction solution was concentrated, and the crude product was purified by silica gel column chromatography to obtain a yellow solid - tert-butyl 2-(5-bromothiophene-2-yl)acetate (6A) (yield 1.6 g, yield 63.81%).

[0200] (2) Preparation of tert-butyl 2-[5-(3-methoxyphenyl)thiophen-2-yl]acetate (6B), the structural formula of which is shown in formula (45).

[0201] To a 100 mL round-bottom flask, tert-butyl 2-(5-bromothiophene-2-yl)acetate (6A) (1.60 g, 5.77 mmol), 3-methoxyphenylboronic acid (1.32 g, 8.66 mmol) and potassium carbonate (2.39 g, 17.32 mmol) were added in sequence, and 1,4-dioxane (30 mL) and water (6 mL) were added respectively, and stirred to dissolve. Then [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium dichloromethane complex (466.74 mg, 0.58 mmol) was added to the solution, and the temperature was raised to 90 ° C under nitrogen protection, and the reaction was carried out for 12 h. After the reaction was completed, the filtrate was collected by filtration, extracted with EA (20 mL × 3), and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate and concentrated. The crude product was purified by silica gel column chromatography to obtain a yellow oily substance - tert-butyl 2-[5-(3-methoxyphenyl)thiophen-2-yl]acetate (6B) (yield 1.15 g, yield rate 65.45%).

[0202] Then, using tert-butyl 2-[5-(3-methoxyphenyl)thiophen-2-yl]acetate (6B) (350 mg, 1.15 mmol) as raw material, a yellow solid, 3-hydroxy-6-{[5-(3-hydroxyphenyl)thiophen-2-yl]methyl}-2-oxo-1H-1,5-naphthyridine-4-carboxamide (yield 6.41 mg, yield rate 6.17%), was prepared in a manner similar to steps (8)-(10) of Example 4.

[0203] The test results are as follows:

[0204] MS (ESI) m / z = 394.1 [M + H] + .

[0205] 1 H NMR (DMSO-d6, 400MHz): δ12.23 (brs, 1H), 10.79 (s, 1H), 9.57 (s, 1H), 8.83 (s, 1H), 8.37 (s, 1H), 7.72-7.61 (m, 1H), 7.35-7.20 (m, 3H), 7.01-6.80 (m, 4H), 4.42 (s, 2H).

[0206] Example 7

[0207] This embodiment provides 3-hydroxy-6-{[4-(1H-pyrrolo[2,3-b]pyridin-3-yl)pyrazol-1-yl]methyl}-2-oxo-1H-1,5-naphthyridine-4-carboxamide. Its structural formula is shown in formula (10). The specific preparation method is as follows, as shown in Table 3.

[0208]

[0209] (1) Preparation of 3-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-7-azaindole (7A), the structural formula of which is shown in formula (46).

[0210] 3-Bromo-7-azaindole (10 g, 50.75 mmol) and DBU (23.18 g, 152.25 mmol) were added to a 500 mL round-bottom flask, and DMF (200 mL) was added and stirred to dissolve. SEM-Cl (10.15 g, 60.90 mmol) was then slowly added dropwise to the solution under an ice-water bath, and the ice-water bath was removed after the addition, and the reaction was continued for 16 h. After the reaction was completed, EA and water were added to separate the mixture, the aqueous phase was extracted with EA, the organic phases were combined, and the organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography to obtain a colorless oil - 3-bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-7-azaindole (7A) (7.6 g, yield 45.75%).

[0211] (2) Preparation of 3-(1H-pyrazol-4-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-7-azaindole (7B), the structural formula of which is shown in formula (47).

[0212] 3-Bromo-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-7-azaindole (7A) (2.5 g, 7.64 mmol) and 1H-pyrazole-4-boric acid (1.28 g, 11.46 mmol) were added to a 250 mL round-bottom flask, 1,4-dioxane (40 mL) was added and stirred to dissolve, and then 8 mL of sodium carbonate aqueous solution (2.43 g, 22.91 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (556.59 mg, 0.76 mmol) were added. The temperature was raised to 90 ° C under nitrogen protection and reacted for 16 h. After the reaction was completed, it was diluted with water, extracted with EA, the organic phases were combined, and the organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography to obtain a brown solid - 3-(1H-pyrazol-4-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-7-azaindole (7B) (yield 276 mg, yield rate 11.49%).

[0213] (3) Preparation of {4-[1-((2-(trimethylsilyl)ethoxy)methyl)-1H-7-azaindol-3-yl]-1H-pyrazol-1-yl}methylboronic acid (7C), the structural formula of which is shown in formula (48).

[0214] 3-(1H-pyrazol-4-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-7-azaindole (7B) (200 mg, 0.64 mmol) and potassium carbonate (264 mg, 1.91 mmol) were added to a 50 mL round-bottom flask, and MeCN (5 mL) was added and stirred at room temperature for 1 h, and then 4 mL of bromomethylboronic acid pinacol ester in MeCN (169 mg, 0.76 mmol) was added, and the reaction was continued at room temperature for 24 h. After the reaction was completed, the mixture was filtered and the filter cake was washed with MeCN to obtain a brown oily substance, {4-[1-((2-(trimethylsilyl)ethoxy)methyl)-1H-7-azaindole-3-yl]-1H-pyrazol-1-yl}methylboronic acid (7C) (yield 210 mg, yield 88.68%).

[0215] Then, using {4-[1-((2-(trimethylsilyl)ethoxy)methyl)-1H-7-azaindol-3-yl]-1H-pyrazol-1-yl}methylboronic acid (7C) (252 mg, 0.677 mmol) as raw material, a yellow solid - 3-hydroxy-6-{[4-(1H-pyrrolo[2,3-b]pyridin-3-yl)pyrazol-1-yl]methyl}-2-oxo-1H-1,5-naphthyridine-4-carboxamide (yield 15 mg, yield 13.81%) was prepared according to a method similar to steps (8)-(10) of Example 4.

[0216] The test results are as follows:

[0217] MS (ESI) m / z = 402.2 [M + H] + .

[0218] 1 H NMR (DMSO-d6, 400MHz): δ12.28 (s, 1H), 11.72 (s, 1H), 10.46 (s, 1H), 8.72 (s, 1H), 8.3 2-8.21 (m, 3H), 7.73-7.68 (m, 1H), 7.31-7.22 (m, 3H), 7.15-6.96 (m, 2H), 5.61 (s, 2H).

[0219] Example 8

[0220] This embodiment provides 3-hydroxy-6-{2-[(4-hydroxyphenyl)amino]-2-oxoethyl}-2-oxo-1H-1,5-naphthyridine-4-carboxamide. Its structural formula is shown in formula (11). The specific preparation method is as follows, as shown in Table 3.

[0221]

[0222] (1) Prepare 2-cyano-2-(8-cyano-7-hydroxy-6-oxo-5H-1,5-naphthyridin-2-yl)acetic acid (8A), the structural formula of which is shown in formula (49).

[0223] To a 50 mL round-bottom flask, tert-butyl cyanoacetate (255 mg, 1.81 mmol), 6-chloro-3-hydroxy-2-oxo-1,2-dihydro-1,5-naphthyridine-4-carbonitrile (4E) (200 mg, 0.90 mmol), lithium tert-butoxide (361 mg, 4.51 mmol), bis(tricyclohexylphosphine)palladium dichloride (67 mg, 0.09 mmol), imid-phos ligand (71 mg, 0.18 mmol) were added, 15 mL of 1,4-dioxane was added to dissolve, the temperature was raised to 130 ° C under nitrogen protection, and the reaction was carried out for 12 hours. After the reaction was completed, the reaction was concentrated, and the crude product was purified by Prep-HPLC to obtain a brown solid - 2-cyano-2-(8-cyano-7-hydroxy-6-oxo-5H-1,5-naphthyridine-2-yl)acetic acid (8A) (yield 487 mg, yield 99.78%).

[0224] (2) Prepare 2-(8-cyano-7-hydroxy-6-oxo-5H-1,5-naphthyridin-2-yl)acetic acid (8B), the structural formula of which is shown in formula (50).

[0225] 2-cyano-2-(8-cyano-7-hydroxy-6-oxo-5H-1,5-naphthyridin-2-yl)acetic acid (8A) (480 mg, 1.8 mmol) was added to a 50 mL round flask, 10 mL of water was added, and then 12 M hydrochloric acid (10 mL) was added, stirred evenly, and then heated to 100 ° C for 3 h. After the reaction was completed, EA was extracted, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography to obtain a yellow oil - 2-(8-cyano-7-hydroxy-6-oxo-5H-1,5-naphthyridin-2-yl)acetic acid (8B) (286 mg, yield 65.50%).

[0226] (3) Prepare 2-(8-cyano-7-hydroxy-6-oxo-5H-1,5-naphthyridin-2-yl)-N-(4-methoxyphenyl)acetamide (8C), the structural formula of which is shown in formula (51).

[0227] To a 50 mL round flask, 2-(8-cyano-7-hydroxy-6-oxo-5H-1,5-naphthyridin-2-yl)acetic acid (8B) (270 mg, 1.1 mmol), 4-anisidine (163 mg, 1.32 mmol) and HOBt (1-hydroxybenzotriazole, 149 mg, 1.1 mmol) were added, 3 mL of DMF was added to dissolve, N,N-diisopropylethylamine (DIPEA, 427 mg, 3.3 mmol) and EDCI (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, 274 mg, 1.43 mmol) were added under an ice-water bath, and the ice-water bath was removed after the addition and the mixture was reacted for 2 h. After the reaction, the reaction mixture was concentrated and the crude product was purified by Prep-HPLC to obtain a yellow solid - 2-(8-cyano-7-hydroxy-6-oxo-5H-1,5-naphthyridin-2-yl)-N-(4-methoxyphenyl)acetamide (8C) (yield 250 mg, yield rate 64.80%).

[0228] (4) Preparation of 3-hydroxy-6-{[4-(4-methoxyphenyl)carboxamide]methyl}-2-oxo-1,2-dihydro-1,5-naphthyridine-4-carboxamide (8D), the structural formula of which is shown in formula (52).

[0229] 2-(8-cyano-7-hydroxy-6-oxo-5H-1,5-naphthyridin-2-yl)-N-(4-methoxyphenyl)acetamide (8C) (250 mg, 0.713 mmol) was added to a 50 mL round flask, and 3 mL of polyphosphoric acid was added at the same time, and the temperature was raised to 110°C and stirred for 1 hour. After the reaction was completed, water was added to quench and filter, and the filter cake was washed with ethanol to obtain a yellow solid 3-hydroxy-6-{[4-(4-methoxyphenyl)formamide]methyl}-2-oxo-1,2-dihydro-1,5-naphthyridine-4-carboxamide (201 mg, yield: 63.72%).

[0230] (5) Preparation of 3-hydroxy-6-{2-[(4-hydroxyphenyl)amino]-2-oxoethyl}-2-oxo-1H-1,5-naphthyridine-4-carboxamide (Example 8)

[0231] 3-Hydroxy-6-{[4-(4-methoxyphenyl)formamide]methyl}-2-oxo-1,2-dihydro-1,5-naphthyridine-4-carboxamide (8D, 100 mg, 0.27 mmol) was added to a 50 mL round flask, 5 mL DCM was added to dissolve, and then 0.68 mL of 2M boron tribromide dichloromethane solution was added under an ice-water bath. After the addition, the ice-water bath was removed and the reaction was continued for 1 hour. After the reaction was completed, the reaction was concentrated, and the crude product was purified by Prep-HPLC to obtain a yellow solid - 3-hydroxy-6-{2-[(4-hydroxyphenyl)amino]-2-oxoethyl}-2-oxo-1H-1,5-naphthyridine-4-carboxamide (yield 5.8 g, yield 5.85%).

[0232] The test results are as follows:

[0233] MS (ESI) m / z = 355.2 [M + H] + .

[0234] 1 H NMR (DMSO-d6, 400MHz): δ12.15 (d, 1H), 10.84 (s, 1H), 10.16-10.00 (m, 2H), 9.24 (d, 1H), 7.74-7.66 (m, 2H), 7.41-7.34 (m, 2H), 7.22-7.09 (m, 2H), 6.97-6.67 (m, 1H), 3.91 (d, 2H).

[0235] Example 9

[0236] This embodiment provides 3-hydroxy-2-oxo-6-{[4-(2-oxo-1H-pyridin-4-yl)pyridin-2-yl]methyl}-1H-1,5-naphthyridine-4-carboxamide, whose structural formula is shown in formula (12). The specific preparation method is as follows, see Table 3 for details.

[0237]

[0238] (1) Preparation of tert-butyl 2-(4-bromopyridin-2-yl)acetate (9A), the structural formula of which is shown in formula (53).

[0239] To a 100 mL round-bottom flask, tert-butyl 2-(4-bromopyridin-2-yl)acetate (4F) (1.5 g, 5.51 mmol), (2-methoxypyridin-4-yl)boric acid (1.01 g, 6.61 mmol), potassium carbonate (1.52 g, 11.02 mmol) were added, and 25 mL of 1,4-dioxane and 5 mL of water were added, respectively, and stirred to dissolve. Then [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (305.57 mg, 0.55 mmol) was added to the solution, and the temperature was raised to 85 ° C under nitrogen protection, and the reaction was carried out for 2 h. After the reaction was completed, the filtrate was collected by filtration and concentrated, and the crude product was purified by silica gel column chromatography to obtain a colorless oily substance-tert-butyl 2-(4-bromopyridin-2-yl)acetate (9A) (yield 1.44 g, yield 86.98%).

[0240] Then, using tert-butyl 2-(4-bromopyridin-2-yl)acetate (9A) (488 mg, 1.62 mmol) as raw material, a method similar to steps (8)-(10) of Example 4 was used to prepare a light yellow solid - 3-hydroxy-2-oxo-6-{[4-(2-oxo-1H-pyridin-4-yl)pyridin-2-yl]methyl}-1H-1,5-naphthyridine-4-carboxamide (yield 13 mg, yield rate 30.23%).

[0241] The test results are as follows:

[0242] MS (ESI) m / z = 390.2 [M + H] + .

[0243] 1 H NMR (DMSO-d6, 400MHz): δ12.00 (br, 1H), 10.70 (s, 1H), 9.98 (s, 1H), 8.70-8.61 (m, 2H), 7.88-7.73 (m, 1H), 7 .66-7.63 (m, 1H), 7.57-7.50 (m, 2H), 7.44-7.42 (m, 1H), 6.77-6.73 (m, 1H), 6.61-6.53 (m, 1H), 4.06 (d, 2H).

[0244] Example 10

[0245] This embodiment provides 3-hydroxy-2-oxo-6-{[5-(4-formamidophenyl)thiophen-2-yl]methyl}-1H-1,5-naphthyridine-4-carboxamide, whose structural formula is shown in formula (13). The specific preparation method is as follows, see Table 3 for details.

[0246]

[0247] (1) Preparation of tert-butyl 2-[5-(4-cyanophenyl)thiophen-2-yl]acetate (10A), the structural formula of which is shown in formula (54).

[0248] Add tert-butyl 2-(5-bromothiophen-2-yl)acetate (2.0 g, 7.22 mmol), 4-cyanophenylboronic acid (1.27 g, 8.66 mmol), potassium carbonate (2.99 g, 21.60 mmol) to a 100 mL round-bottom flask, add 20 mL of 1,4-dioxane and 4 mL of water, respectively, and stir to dissolve. Then add [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (589 mg, 0.72 mmol) to the solution, heat to 90 ° C under nitrogen protection, and react for 2 h. After the reaction is completed, filter and collect the filtrate and concentrate it. The crude product is purified by silica gel column chromatography to obtain a yellow oily substance - tert-butyl 2-[5-(4-cyanophenyl)thiophen-2-yl]acetate (10A) (yield 1.30 g, yield 60.00%).

[0249] Then, using tert-butyl 2-(4-bromopyridin-2-yl)acetate (10A) (405 mg, 1.35 mmol) as raw material, a method similar to steps (8)-(10) of Example 4 was used to prepare a light yellow solid - 3-hydroxy-2-oxo-6-{[5-(4-formamidophenyl)thiophen-2-yl]methyl}-1H-1,5-naphthyridine-4-carboxamide (yield 5.47 mg, total yield 1.30%).

[0250] The test results are as follows:

[0251] MS (ESI) m / z = 421.0 [M + H] + .

[0252] 1 H NMR (DMSO-d6, 400MHz): δ 10.78 (s, 1H), 9.96 (s, 1H), 8.84 (s, 1H), 7.98-7.66 (m, 5H), 7.48-6.67 (m, 6H), 4.49 (s, 2H).

[0253] Embodiment 11

[0254] This embodiment provides 3-hydroxy-2-oxo-6-{[3-(4-formamidophenyl)phenyl]methyl}-1H-1,5-naphthyridine-4-carboxamide. Its structural formula is shown in formula (14). The specific preparation method is as follows, as shown in Table 3.

[0255]

[0256] (1) Preparation of tert-butyl 2-(3-bromophenyl)acetate (11A), the structural formula of which is shown in formula (55).

[0257] In a 100 mL round-bottom flask, (3-bromophenyl)acetic acid (5.0 g, 23.3 mmol) and DMAP (1.42 g, 11.6 mmol) were added, and 50 mL of tert-butyl alcohol and 50 mL of THF were added respectively, and the mixture was stirred to dissolve. Then Boc anhydride (10.2 g, 46.50 mmol) was added to the solution, and the mixture was reacted at room temperature for 12 h. After the reaction was completed, the filtrate was collected by filtration and concentrated, and the crude product was purified by silica gel column chromatography to obtain a yellow oily substance, tert-butyl 2-(3-bromophenyl)acetate (11A) (yield 5.80 g, yield 92.00%).

[0258] (2) Preparation of tert-butyl 2-[3-(4-cyanophenyl)phenyl]acetate (11B), the structural formula of which is shown in formula (56).

[0259] To a 100 mL round-bottom flask, tert-butyl 2-(3-bromophenyl)acetate (11A) (3.0 g, 11.1 mmol), 4-cyanophenylboronic acid (2.11 g, 14.4 mmol), potassium carbonate (4.59 g, 33.2 mmol), Pd(dppf)Cl2 (catalyst) (809 mg, 1.11 mmol) were added, and 40 mL of 1,4-dioxane was added and stirred to dissolve. The temperature was raised to 90 ° C under nitrogen protection and reacted for 12 h. After the reaction was completed, EA was extracted, and the organic phase was washed with saturated brine, and the organic phase was dried over anhydrous sodium sulfate and concentrated. The crude product was purified by silica gel column chromatography to obtain a white solid - tert-butyl 2-[3-(4-cyanophenyl)phenyl]acetate (11B) (yield 3.2 g, yield 98.00%).

[0260] Then, using tert-butyl 2-[3-(4-cyanophenyl)phenyl]acetate (11B) (298 mg, 1.02 mmol) as raw material, a method similar to steps (8)-(10) of Example 4 was used to prepare a yellow solid - 3-hydroxy-2-oxo-6-{[3-(4-formamidophenyl)phenyl]methyl}-1H-1,5-naphthyridine-4-carboxamide (yield 91.18 mg, total yield 21.57%).

[0261] The test results are as follows:

[0262] MS (ESI) m / z = 415.1 [M + H] + .

[0263] 1H NMR (DMSO-d6, 400MHz): δ 11.99 (s, 1H), 10.73 (s, 1H), 9.96 (s, 1H), 8.03-7.95 (m, 3H), 7.80-7.57 (m, 9H), 7.50-7.18 (m, 1H), 4.31 (s, 2H).

[0264] Example 12

[0265] This embodiment provides 3-hydroxy-2-oxo-6-{[4-(5-carboxamido-1H-pyrrol-3-yl)pyrazol-1-yl]methyl}-1H-1,5-naphthyridine-4-carboxamide. Its structural formula is shown in formula (15). The specific preparation method is as follows, as shown in Table 3.

[0266]

[0267] (1) Preparation of 4-bromo-1-[(2-trimethylsilyl)ethoxy]methyl-1H-pyrrole-2-carboxylic acid methyl ester (12A), the structural formula of which is shown in formula (57).

[0268] 4-bromo-1H-pyrrole-2-carboxylic acid methyl ester (10.0 g, 49.0 mmol) and DBU (22.4 g, 147 mmol) were added to a 100 mL round-bottom flask, and 150 mL of DCM was added and stirred to dissolve. SEM-Cl (12.3 g, 73.5 mmol) was added dropwise under an ice-water bath, and the mixture was reacted at room temperature for 12 h. After the reaction was completed, it was diluted with water, extracted with DCM, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography to obtain a colorless oil - 4-bromo-1-[(2-trimethylsilyl)ethoxy]methyl-1H-pyrrole-2-carboxylic acid methyl ester (12A) (yield 9.70 g, yield 59.00%).

[0269] (2) Preparation of 4-(1H-pyrazol-4-yl)-1-[(2-trimethylsilyl)ethoxy]methyl-1H-pyrrole-2-carboxylic acid methyl ester (12B), the structural formula of which is shown in formula (58).

[0270] 4-Bromo-1-[(2-trimethylsilyl)ethoxy]methyl-1H-pyrrole-2-carboxylic acid methyl ester (12A) (7.5 g, 22.4 mmol), 1H-pyrazole-4-pinacol borate (6.53 g, 33.7 mmol), potassium carbonate (9.30 g, 67.3 mmol), Pd(dppf)Cl2 (catalyst) (1.64 g, 2.24 mmol) were added to a 100 mL round-bottom flask, and 140 mL DMF and 35 mL water were added to stir and dissolve, and the temperature was raised to 80 ° C under nitrogen protection, and the reaction was carried out for 12 hours. After the reaction was completed, it was diluted with water, extracted with EA, and the organic phase was washed with saturated brine, and the organic phase was dried over anhydrous sodium sulfate and concentrated. The crude product was purified by silica gel column chromatography to obtain a yellow oily substance - 4-(1H-pyrazol-4-yl)-1-[(2-trimethylsilyl)ethoxy]methyl-1H-pyrrole-2-carboxylic acid methyl ester (12B) (yield 2.60 g, yield rate 36.00%).

[0271] (3) Prepare 4-[1-(boronic acid methyl)pyrazol-4-yl]-1-(2-trimethylsilyl)ethoxy)methyl-1H-pyrrole-2-carboxylic acid methyl ester (12C), the structural formula of which is shown in formula (59).

[0272] 4-(1H-pyrazol-4-yl)-1-[(2-trimethylsilyl)ethoxy]methyl-1H-pyrrole-2-carboxylic acid methyl ester (12B) (2.6 g, 8.09 mmol), potassium carbonate (3.35 g, 24.3 mmol) were added to a 100 mL round-bottom flask, 50 mL of acetonitrile was added and stirred to dissolve, and then bromomethylbenzoic acid pinacol (2.14 g, 9.71 mmol) was added. The reaction was allowed to proceed at room temperature for 12 h. After the reaction was completed, the filtrate was collected by filtration and concentrated, and the crude product was purified by silica gel column chromatography to obtain a yellow oily substance - 4-[1-(boronic acid methyl)pyrazol-4-yl]-1-(2-trimethylsilyl)ethoxy)methyl-1H-pyrrole-2-carboxylic acid methyl ester (12C) (yield 3.0 g, yield 97.00%).

[0273] Then, using 4-[1-(boronic acid methyl) pyrazol-4-yl]-1-(2-trimethylsilyl) ethoxy) methyl-1H-pyrrole-2-carboxylic acid methyl ester (12C) (385 mg, 1.02 mmol) as raw material, a method similar to steps (8)-(10) of Example 4 was used to prepare a light yellow solid - 3-hydroxy-2-oxo-6-{[4-(5-formamido-1H-pyrrol-3-yl) pyrazol-1-yl] methyl}-1H-1,5-naphthyridine-4-carboxamide (yield 4.59 mg, yield rate 6.0%).

[0274] The test results are as follows:

[0275] MS (ESI) m / z = 394.1 [M + H] + .

[0276] 1 H NMR (DMSO-d6, 400MHz): δ11.38(s, 1H), 10.51(s, 1H), 8.75(s, 1H), 8.17(s, 1H), 7.92-7.90(m, 1H), 7.69-7.46(m, 5H), 7.06-6.87(m, 3H), 5.53(s, 2H).

[0277] Embodiment 13

[0278] This embodiment provides 3-hydroxy-2-oxo-6-{[6-(1H-pyrrolo[2,3-b]pyridin-3-yl)pyridin-3-yl]methyl-1H-1,5-naphthyridine-4-carboxamide. Its structural formula is shown in formula (16). The specific preparation method is as follows, as shown in Table 3.

[0279]

[0280] (1) Prepare 3-bromo-1-[(2-trimethylsilyl)ethoxy]methyl-1H-pyrrolo[2,3-b]pyridine (13A), the structural formula of which is shown in formula (60).

[0281] 3-Bromo-1H-pyrrolo[2,3-b]pyridine (30 g, 156 mmol) and DBU (70 g, 457 mmol) were added to a 100 mL round-bottom flask, and 300 mL THF was added and stirred to dissolve. SEM-Cl (38 g, 228 mmol) was added dropwise under an ice-water bath, and the mixture was reacted at room temperature for 16 h. After the reaction was completed, the filtrate was collected by filtration and concentrated, and the crude product was purified by silica gel column chromatography to obtain a colorless oil - 3-bromo-1-[(2-trimethylsilyl)ethoxy]methyl-1H-pyrrolo[2,3-b]pyridine (13A) (yield 21 g, yield 42%).

[0282] (2) Prepare 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[(2-trimethylsilyl)ethoxy]methyl-1H-pyrrolo[2,3-b]pyridine (13B), the structural formula of which is shown in formula (61).

[0283] 3-Bromo-1-[(2-trimethylsilyl)ethoxy]methyl-1H-pyrrolo[2,3-b]pyridine (13A) (19.2 g, 58.6 mmol), biboronic acid pinacol ester (22.3 g, 88.0 mmol), potassium acetate (17.3 g, 199.4 mmol), Pd(dppf)Cl2 (catalyst) (5.3 g, 7.2 mmol) were added to a 100 mL round-bottom flask, and 240 mL DMF and 60 mL water were added to stir and dissolve, and the temperature was raised to 80 ° C under nitrogen protection, and the reaction was carried out for 12 hours. After the reaction was completed, it was diluted with water, extracted with EA, and the organic phase was washed with saturated brine, and the organic phase was dried over anhydrous sodium sulfate and concentrated. The crude product was purified by silica gel column chromatography to obtain a yellow oil - 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-[(2-trimethylsilyl)ethoxy]methyl-1H-pyrrolo[2,3-b]pyridine (13B) (yield 11.9 g, yield rate 78.00%).

[0284] (3) Preparation of tert-butyl 2-(6-chloropyridin-3-yl)acetate (13C), the structural formula of which is shown in formula (62).

[0285] 2-(6-chloropyridin-3-yl)acetic acid (5.0 g, 29.14 mmol) and DMAP (1.78 g, 14.57 mmol) were added to a 100 mL round-bottom flask, and 50 mL of THF and 50 mL of tert-butyl alcohol were added to stir and dissolve, and then Boc anhydride (12.72 g, 58.28 mmol) was added under an ice-water bath, and the reaction was carried out at room temperature for 6 h. After the reaction was completed, the filtrate was collected by filtration and concentrated, and the crude product was purified by silica gel column chromatography to obtain a light yellow oily substance - tert-butyl 2-(6-chloropyridin-3-yl)acetate (13C) (yield 6.3 g, yield 95.00%).

[0286] (4) Preparation of tert-butyl 2-{6-[(1-(2-trimethylsilyl)ethoxy)methyl-1H-pyrrolo[2,3-b]pyridin-3-yl]pyridin-3-yl}acetate (13D), the structural formula of which is shown in formula (63).

[0287] 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-1-[(2-trimethylsilyl)ethoxy]methyl-1H-pyrrolo[2,3-b]pyridine (13B) (7.4 g, 19.76 mmol) and tert-butyl 2-(6-chloropyridin-3-yl)acetate (13C) (3.0 g, 13.18 mmol) were added to a 100 mL round-bottom flask, and 100 mL of 1,4-dioxane and 20 mL of water were added to stir and dissolve, and then potassium carbonate (5.46 g, 39.53 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (964.07 mg, 1.32 mmol) were added, and the temperature was raised to 90°C under nitrogen protection for 16 h. After the reaction was completed, it was diluted with water, extracted with EA, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography to obtain a light yellow oil - tert-butyl 2-{6-[(1-(2-trimethylsilyl)ethoxy)methyl-1H-pyrrolo[2,3-b]pyridin-3-yl]pyridin-3-yl}acetate (13D) (yield 2.4 g, yield rate 41.00%).

[0288] Then, using tert-butyl 2-{6-[(1-(2-trimethylsilyl)ethoxy)methyl-1H-pyrrolo[2,3-b]pyridin-3-yl]pyridin-3-yl}acetate (13D) (505.89 mg, 1.15 mmol) as raw material, a method similar to steps (8)-(10) of Example 4 was used to prepare a yellow solid - 3-hydroxy-2-oxo-6-{[6-(1H-pyrrolo[2,3-b]pyridin-3-yl)pyridin-3-yl]methyl-1H-1,5-naphthyridine-4-carboxamide (yield 17.3 mg, total yield 3.65%).

[0289] The test results are as follows:

[0290] MS (ESI) m / z = 413.1 [M + H] + .

[0291] 1 H NMR (DMSO-d6, 400MHz): δ11.99 (s, 1H), 10.75 (s, 1H), 9.95 (s, 1H), 8.75-8.55 (m, 2H) , 8.26-8.18(m, 2H), 7.89-7.65(m, 5H), 7.42-7.16(m, 2H), 6.54(s, 1H), 4.27(s, 2H).

[0292] Embodiment 14

[0293] This embodiment provides 3-hydroxy-2-oxo-6-{[5-(1H-pyrrolo[2,3-b]pyridin-3-yl)thiophen-2-yl]methyl}-1H-1,5-naphthyridine-4-carboxamide. Its structural formula is shown in formula (17). The specific preparation method is as follows, as shown in Table 3.

[0294]

[0295] (1) Preparation of tert-butyl 2-{5-[(1-(2-trimethylsilyl)ethoxy)methyl-1H-pyrrolo[2,3-b]pyridin-3-yl]thiophen-2-yl}acetate (14A), the structural formula of which is shown in formula (64).

[0296] 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-1-[(2-trimethylsilyl)ethoxy]methyl-1H-pyrrolo[2,3-b]pyridine (13B) (3.0 g, 8.01 mmol), tert-butyl 2-(5-bromothiophen-2-yl)acetate (2.2 g, 8.01 mmol), potassium phosphate (5.1 g, 24.04 mmol) were added to a 100 mL round-bottom flask, and 30 mL of 1,4-dioxane and 6 mL of water were added respectively, and stirred to dissolve. Then [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (293.18 mg, 0.53 mmol) was added to the solution, and the temperature was raised to 100 ° C under nitrogen protection, and the reaction was carried out for 10 hours. After the reaction, the organic phase was collected and concentrated, and the crude product was purified by silica gel column chromatography to obtain a yellow oil - tert-butyl 2-{5-[(1-(2-trimethylsilyl)ethoxy)methyl-1H-pyrrolo[2,3-b]pyridin-3-yl]thiophen-2-yl}acetate (14A) (yield 600 mg, yield 16.84%).

[0297] Then, using tert-butyl 2-{5-[(1-(2-trimethylsilyl)ethoxy)methyl-1H-pyrrolo[2,3-b]pyridin-3-yl]thiophen-2-yl}acetate (14A) (231 mg, 0.52 mmol) as raw material, a method similar to steps (8)-(10) of Example 4 was used to prepare a light yellow solid - 3-hydroxy-2-oxo-6-{[5-(1H-pyrrolo[2,3-b]pyridin-3-yl)thiophen-2-yl]methyl}-1H-1,5-naphthyridine-4-carboxamide (yield 5.64 mg, total yield 2.60%).

[0298] The test results are as follows:

[0299] MS (ESI) m / z = 418.1 [M + H] + .

[0300] 1 H NMR (DMSO-d6, 400MHz): δ11.93 (s, 1H), 10.85 (s, 1H), 9.96 (s, 1H), 8.90 (s, 1H), 8.27-8.21(m, 2H), 7.78-7.67(m, 2H), 7.27-6.96(m, 5H), 4.46(s, 2H).

[0301] Embodiment 15

[0302] This embodiment provides 3-hydroxy-2-oxo-6-({5-[3-(4-methylpiperazin-1-yl)phenyl]thiophen-2-yl}methyl)-1H-1,5-naphthyridine-4-carboxamide. Its structural formula is shown in formula (18). The specific preparation method is as follows, see Table 3 for details.

[0303]

[0304] (1) Preparation of 1-(3-bromophenyl)-4-methylpiperazine (15A), the structural formula of which is shown in formula (65).

[0305] 1-(3-bromophenyl)piperazine (9.50 g, 34.22 mmol) and formaldehyde (4.17 g, 51.33 mmol) were added to a 100 mL round-bottom flask, and 100 mL of dichloroethane was added to stir and dissolve. Octadecyltrimethylammonium bromide (10.88 g, 51.33 mmol) was then added to the solution, and the reaction was continued at room temperature for 16 h. After the reaction was completed, saturated sodium carbonate solution was added to adjust the pH to 9, and the organic phase was separated and dried by spin drying. The crude product was purified by silica gel column chromatography to obtain a colorless oily substance, 1-(3-bromophenyl)-4-methylpiperazine (15A) (yield 8.50 g, yield 97.00%).

[0306] (2) Prepare 1-methyl-4-[3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]piperazine (15B), the structural formula of which is shown in formula (66).

[0307] 1-(3-bromophenyl)-4-methylpiperazine (15A) (8.50 g, 33.31 mmol), biboronic acid pinacol ester (16.92 g, 66.62 mmol), potassium acetate (9.81 g, 99.94 mmol), Pd(dppf)Cl2 (catalyst) (1.22 g, 1.67 mmol) were added to a 100 mL round-bottom flask, and 250 mL of 1,4-dioxane was added and stirred to dissolve, and the temperature was raised to 90 ° C under nitrogen protection, and the reaction was carried out for 12 hours. After the reaction was completed, it was diluted with water, extracted with EA, and the organic phase was washed with saturated brine, and the organic phase was dried over anhydrous sodium sulfate and concentrated. The crude product was purified by silica gel column chromatography to obtain a black oily substance - 1-methyl-4-[3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl]piperazine (15B) (yield 7.60 g, yield rate 75.00%).

[0308] (3) Preparation of tert-butyl {5-[(3-(4-methylpiperazin-1-yl)phenyl]thiophen-2-yl}acetate (15C), the structural formula of which is shown in formula (67).

[0309] 1-Methyl-4-[3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)phenyl]piperazine (15B) (3.05 g, 10.10 mmol) and tert-butyl (5-thiophen-2-yl)acetate (2.80 g, 10.10 mmol) were added to a 100 mL round-bottom flask, and 50 mL of 1,4-dioxane and 10 mL of water were added to stir and dissolve, and then potassium carbonate (4.19 g, 30.31 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (824.95 mg, 1.01 mmol) were added, and the temperature was raised to 90°C under nitrogen protection to react for 16 h. After the reaction was completed, it was diluted with water, extracted with EA, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography to obtain a black oily substance - tert-butyl {5-[(3-(4-methylpiperazin-1-yl)phenyl]thiophen-2-yl}acetate (15C) (yield 2.4 g, yield rate 64.00%).

[0310] Then, using tert-butyl {5-[(3-(4-methylpiperazin-1-yl)phenyl]thiophen-2-yl}acetate (15C) (1.13 g, 1.02 mmol) as raw material, a yellow solid, 3-hydroxy-2-oxo-6-({5-[3-(4-methylpiperazin-1-yl)phenyl]thiophen-2-yl}methyl)-1H-1,5-naphthyridine-4-carboxamide, was prepared in a manner similar to steps (8)-(10) of Example 4 (yield 19.70 mg, total yield 4.07%).

[0311] The test results are as follows:

[0312] MS (ESI) m / z = 476.1 [M + H] + .

[0313] 1 H NMR (DMSO-d6, 400MHz): δ12.28 (s, 1H), 10.78 (s, 1H), 7.78-7.72 (m, 3H), 7.43-7. 18(m, 3H), 7.07-6.85(m, 4H), 4.45(s, 2H), 3.16(t, 4H), 2.45(t, 4H), 2.22(s, 3H).

[0314] Example 16

[0315] This embodiment provides 4-{3-[(8-formamido-7-hydroxy-6-oxo-5H-1,5-naphthyridin-2-yl)methyl]phenyl}benzoic acid. Its structural formula is shown in formula (19). The specific preparation method is as follows, as shown in Table 3.

[0316]

[0317] (1) Preparation of 3'-methyl-[1,1'-biphenyl]-4-carboxylic acid methyl ester (16A), the structural formula of which is shown in formula (68).

[0318] 3-iodotoluene (5.0 g, 22.9 mmol), methyl 4-boronic acid benzoate (4.95 g, 27.5 mmol), potassium carbonate (9.51 g, 68.8 mmol) were added to a 100 mL round-bottom flask, and 50 mL of 1,4-dioxane and 10 mL of water were added and stirred to dissolve. Then [1,1'-bis(diphenylphosphino)ferrocene] palladium dichloride (1.68 g, 2.29 mmol) was added to the solution, and the temperature was raised to 90 ° C under nitrogen protection, and the reaction was carried out for 2 h. After the reaction was completed, it was diluted with water, extracted with EA, and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The crude product was purified by silica gel column chromatography to obtain a yellow solid - 3'-methyl-[1,1'-biphenyl]-4-carboxylic acid methyl ester (16A) (yield 4.9 g, yield 94%).

[0319] (2) Preparation of 3'-bromomethyl-[1,1'-biphenyl]-4-carboxylic acid methyl ester (16B), the structural formula of which is shown in formula (69).

[0320] 3'-Methyl-[1,1'-biphenyl]-4-carboxylic acid methyl ester (16A) (4.8 g, 21.2 mmol), NBS reagent (4.15 g, 23.3 mmol) were added to a 100 mL round-bottom flask, and 60 mL of carbon tetrachloride was added, and then dibenzoyl peroxide (0.51 mg, 2.12 mmol) was added to the solution, and the temperature was raised to 80°C and reacted for 12 hours. After the reaction was completed, the filtrate was collected by filtration and concentrated, and the crude product was purified by silica gel column chromatography to obtain a white solid - 3'-bromomethyl-[1,1'-biphenyl]-4-carboxylic acid methyl ester (16B) (yield 4.6 g, yield 89%).

[0321] (3) Preparation of 3'-[(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)]methyl-[1,1'-biphenyl]-4-carboxylic acid methyl ester (16C), the structural formula of which is shown in formula (70).

[0322] 3'-bromomethyl-[1,1'-biphenyl]-4-carboxylic acid methyl ester (16B) (4.5 g, 14.8 mmol), biboronic acid pinacol ester (4.49 g, 17.7 mmol), potassium acetate (4.34 g, 44.2 mmol), Pd(dppf)Cl2 (catalyst) (1.08 g, 1.47 mmol) were added to a 100 mL round-bottom flask, and 90 mL of 1,4-dioxane was added and stirred to dissolve, and the temperature was raised to 80°C under nitrogen protection, and the reaction was continued for 2 hours. After the reaction was completed, the filtrate was collected by filtration and concentrated, and the crude product was purified by silica gel column chromatography to obtain a white solid - 3'-[(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)]methyl-[1,1'-biphenyl]-4-carboxylic acid methyl ester (16C) (yield 2.8 g, yield 53%).

[0323] Then, using 3'-[(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)]methyl-[1,1'-biphenyl]-4-carboxylic acid methyl ester (16C) (405 mg, 1.15 mmol) as raw material, a yellow solid - 4-{3-[(8-formamido-7-hydroxy-6-oxo-5H-1,5-naphthyridin-2-yl)methyl]phenyl}benzoic acid (yield 9.87 mg, total yield 2.07%) was prepared according to a method similar to steps (8)-(10) of Example 4.

[0324] The test results are as follows:

[0325] MS (ESI) m / z = 416.2 [M + H] + .

[0326] 1H NMR (DMSO-d6, 400MHz): δ 11.95 (s, 1H), 10.72 (s, 1H), 9.96 (s, 1H), 8.72 (s, 1H), 8.03-8.01 (m, 2H), 7.82-7.21 (m, 9H), 4.31 (s, 2H).

[0327] Embodiment 17

[0328] This embodiment provides 3-hydroxy-2-oxo-6-({3-[(4-hydroxyphenyl)methyl]phenyl}methyl)-1H-1,5-naphthyridine-4-carboxamide. Its structural formula is shown in formula (20), and its preparation method is as follows, as shown in Table 3.

[0329]

[0330] (1) Preparation of [3-(4-methoxybenzyl)phenyl]methanol (17A), the structural formula of which is shown in formula (71).

[0331] To a 100 mL round-bottom flask, add (3-bromophenyl)methanol (2.5 g, 13.3 mmol), 4-methoxybenzylboronic acid pinacol ester (3.98 g, 16.0 mmol), potassium carbonate (5.54 g, 40.1 mmol), add 50 mL of 1,4-dioxane and 10 mL of water, respectively, and stir to dissolve. Then add [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (978 mg, 1.34 mmol) to the solution, heat to 90 ° C under nitrogen protection, and react for 12 h. After the reaction is completed, dilute with water, extract with EA, wash the organic phase with saturated brine, dry the organic phase with anhydrous sodium sulfate, and concentrate. The crude product is purified by silica gel column chromatography to obtain a yellow oily substance, [3-(4-methoxybenzyl)phenyl]methanol (17A) (yield 1.60 g, yield 52%).

[0332] (2) Preparation of 1-[3-(4-methoxybenzyl)]benzyl bromide (17B), the structural formula of which is shown in formula (72).

[0333] Triphenylphosphine (2.59 g, 9.86 mmol) was added to a 100 mL round-bottom flask, and 50 mL of THF was added and stirred to dissolve. Carbon tetrabromide (3.27 g, 9.86 mmol) and [3-(4-methoxybenzyl)phenyl]methanol (17A) (1.50 g, 6.57 mmol) were slowly added under an ice-water bath and dissolved in 10 mL of THF. The mixture was slowly added dropwise to the above solution under an ice-water bath, and the mixture was reacted at room temperature for 2 h. After the reaction was completed, the filtrate was collected by filtration and concentrated, and the crude product was purified by silica gel column chromatography to obtain a yellow oily substance, 1-[3-(4-methoxybenzyl)]benzyl bromide (17B) (yield 1.70 g, yield 89%).

[0334] (3) Prepare 2-[3-(4-methoxybenzyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (17C), the structural formula of which is shown in formula (73).

[0335] 1-[3-(4-methoxybenzyl)]benzyl bromide (17B) (1.6 g, 5.49 mmol), bipyraclostrobin (1.67 g, 6.59 mmol), potassium acetate (1.62 g, 16.5 mmol) were added to a 100 mL round-bottom flask, and 30 mL of 1,4-dioxane was added and stirred to dissolve. Then [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (402 mg, 0.55 mmol) was added to the solution, and the temperature was raised to 80°C under nitrogen protection, and the reaction was carried out for 12 hours. After the reaction was completed, the filtrate was collected by filtration and concentrated, and the crude product was purified by silica gel column chromatography to obtain a yellow oily substance - 2-[3-(4-methoxybenzyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborane (17C) (yield 1.60 g, yield 86%).

[0336] Then, using 2-[3-(4-methoxybenzyl)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (17C) (366 mg, 1.08 mmol) as raw material, a yellow solid, 3-hydroxy-2-oxo-6-({3-[(4-hydroxyphenyl)methyl]phenyl}methyl)-1H-1,5-naphthyridine-4-carboxamide, was prepared in a manner similar to steps (8)-(10) of Example 4 (yield 22.2 mg, total yield 5.1%).

[0337] The test results are as follows:

[0338] MS (ESI) m / z = 402.2 [M + H] + .

[0339] 1 H NMR (DMSO-d6, 400MHz): δ12.03 (br, 1H), 10.73 (s, 1H), 9.17 (s, 1H), 7.78-7 .61 (m, 3H), 7.34-6.99 (m, 7H), 6.65-6.63 (m, 2H), 4.16 (s, 2H), 3.78 (s, 2H).

[0340] The results of biological tests on the cyclic α-hydroxyimide compounds are described in detail below to further illustrate the technical solution of the present application.

[0341] Test 1

[0342] The test subjects are the cyclic α-hydroxyimide compounds provided in Examples 1-17 of the present application, and the test type is to detect the inhibitory ability of the above compounds on FEN-1.

[0343] The details are as follows:

[0344] (1) Test method

[0345] A. Expression and purification of wild-type human FEN-1 protein

[0346] The wild-type human FEN-1 protein expression system was constructed using the E. coli BL21 (DE3) expression system (Vazyme BL21 (DE3) competent cells), and the purified FEN-1 protein was obtained by the E. coli BL21 (DE3) expression system. The FEN-1 plasmid was synthesized by Genscript (UniProt Entry: P39748).

[0347] B. Construction of FEN-1 enzyme activity detection system based on fluorescence signal

[0348] Construction of DNA substrate containing HBV sequence: DNA substrate with HBV r sequence 5'-flap structure, connected with fluorescence generating group and fluorescence quenching group structure, prepared by annealing "flap", "quencher" and "template" oligonucleotide sequences (F is 5-TAMRA fluorescence group, Q represents BHQ fluorescence quenching group). The specific sequences are shown in Table 6.

[0349] Table 6 Construction of DNA substrate containing HBV sequence

[0350]

[0351] The annealing program was as follows: 95°C, 5 min; then cooled to 25°C.

[0352] The annealed DNA substrate was co-incubated with the purified FEN-1 protein (100 nM). The F fragment cleaved by FEN-1 emits fluorescence because it is separated from the Q fragment. Therefore, the increased fluorescence intensity can indicate the cleavage activity of FEN-1 on the "flap". Different concentration gradients of the compounds to be tested (compounds provided in Examples 1-17) were added to this system, and the fluorescence change data were collected by the fluorescence detection equipment. Inhibition rate and IC 50 The calculation formula of (half inhibitory concentration) is as follows:

[0353] Inhibition rate and IC 50 The calculation formula is as follows:

[0354]

[0355] IC 50 of the compound is calculated by XL fit software-Fit models(205).

[0356] (2) Test results

[0357] The test results are shown in Table 7. 50 (half inhibitory concentration) indicates the concentration of the compound to be tested when it inhibits half the amount of FEN-1 protein; the stronger the inhibitory ability, the lower the concentration;

[0358] Table 7 Activity data of cyclic α-hydroxyimide compounds provided in Examples 1-17 for inhibiting FEN-1

[0359]

[0360]

[0361] It can be seen from Table 7 that the cyclic α-hydroxyimide compound provided by the present application can effectively inhibit the activity of FEN-1 protein.

[0362] Test 2

[0363] The test subject is the cyclic α-hydroxyimide compound provided in Example 13 of the present application, and the test type is a functional test to detect the compound on PHH cells (primary human hepatocytes) to evaluate the in vitro anti-HBV efficacy of the compound provided in the present application.

[0364] The test method is as follows:

[0365] (1) Test method

[0366] On day 0, thaw the frozen PHH cells and adjust the density to 6×10 5 PHH cells were plated into 24-well plates with 0.45 ml per well.

[0367] On day 1, HBV subtype D was added to infect PHH cells.

[0368] On day 2, compounds were added to treat PHH cells.

[0369] The test compound had a starting concentration of 50 μM, 4-fold dilution, 5 concentrations, and duplicate wells. The reference compound GLS4 (a new drug for hepatitis B) had a starting concentration of 5 μM, 4-fold dilution, 5 concentrations, and duplicate wells. The fresh medium containing the compound was replaced every 2 days. The final concentration of DMSO (dimethyl sulfoxide) in the medium was 2%.

[0370] On the 8th day, the supernatant of PHH cells was collected in a 24-well plate, and HBeAg (hepatitis B e antigen) in the supernatant of PHH cells was detected by ELISA, and the fluorescence change data was collected by a fluorescence detection device.

[0371] The viability of PHH cells was determined by CCK-8 method. PHH cells were collected, Hirt DNA in PHH cells was extracted, and cccDNA was detected by Southern blot (two duplicate wells were combined as one well for the test).

[0372] (2) Test results

[0373] The inhibitory activity data are shown in Table 8. Among them, EC 50 (Median lethal concentration) indicates the concentration of the compound to be tested that causes 50% PHH cell death, poisoning or pathology; the higher the concentration, the safer the compound to be tested is for PHH cells. 50 (Half inhibitory concentration) indicates the concentration of the compound to be tested that causes 50% PHH cell death, poisoning or pathology; the stronger the inhibitory ability, the lower the concentration.

[0374] Table 8 Inhibitory activity data of Example 13 on HBeAg

[0375] Compound Type <![CDATA[HBeAg EC 50 (μM)]]> <![CDATA[CC 50 (μM)]]> Example 13 provides the compound 21.45 >50

[0376] The inhibition rate of HbeAg by the tested compound on the 8th day is as follows: Figure 3 The results of Southern blot detection of cccDNA are shown in Figure 4 shown.

[0377] Depend on Figure 3 , Figure 4 As shown in Table 8, the compound provided in Example 13 of the present application can not only significantly reduce the production of HBeAg in PHH cells, but also inhibit the formation of cccDNA, showing the potential to cure hepatitis B.

[0378] This specific embodiment is merely an explanation of the present application and is not a limitation of the present application. After reading this specification, those skilled in the art may make modifications to the present embodiment without any creative contribution as needed, but such modifications are protected by the patent law as long as they are within the scope of the claims of the present application.

Claims

1. A cyclic α-hydroxyimide compound, characterized in that The general structural formula of the cyclic α-hydroxyimide compound is as follows: In the general structural formula, R1 is hydrogen, C 1-6 Alkyl, C 3-7 Cycloalkyl, C 3-7 Heterocycloalkyl, aryl or heteroaryl substituted C 1-6 Any of the alkyl groups; R2 is hydrogen, deuterium, C 1-6 Alkyl, C 3-7 Cycloalkyl, C 3-7 Heterocycloalkyl, C 1-6 Alkoxy, C 1-6 any of alkylamino, optionally substituted aryl or optionally substituted heteroaryl; X is any of CH and N; Y is any one of hydrogen, deuterium, halogen, hydroxyl, amino, cyano, and a group as shown in formula (2); In the formula (1), R3 is hydrogen, hydroxyl, amino, C 1-6 Alkoxy, C 1-6 Any of alkylamino groups.

2. The cyclic α-hydroxyimide compound according to claim 1, characterized in that The structural formula of the cyclic α-hydroxyimide compound is shown in formula (2) or formula (3): R4 is hydrogen, deuterium, C 1-6 Alkyl, C 3-7 Cycloalkyl, C 3-7 Heterocycloalkyl, C 1-6 Alkoxy, C 1-6 any of alkylamino, optionally substituted aryl or optionally substituted heteroaryl; Z is any one of CH2, NH, O and S.

3. The cyclic α-hydroxyimide compound according to claim 1, characterized in that In the general structural formula, R1 is a C substituted by an aryl or heteroaryl group. 1-6 alkyl; R2 is optionally substituted aryl or optionally substituted heteroaryl; X is any of CH and N; Y is a group as shown in formula (1); in formula (1), R3 is an amino group or a C 1-6 Alkoxy; Z is CH2.

4. The cyclic α-hydroxyimide compound according to claim 1, characterized in that The structural formula of the cyclic α-hydroxyimide compound is any one of formula (4) to formula (20):

5. Use of the cyclic α-hydroxyimide compound according to any one of claims 1 to 4 in the preparation of a medicament for preventing and treating hepatitis B virus infection.

6. A composition, characterized in that The composition comprises the cyclic α-hydroxyimide compound according to any one of claims 1 to 4; Preferably, the composition is a preparation; the preparation further comprises a pharmaceutically acceptable salt; Preferably, the pharmaceutically acceptable salts include acid addition salts and base addition salts.

7. A solvate, characterized in that The solvate is an association complex formed by the cyclic α-hydroxyimide compound according to any one of claims 1 to 4 and one or more solvent molecules; preferably, the solvent includes any one or more of water, methanol, ethanol, isopropanol, ethyl acetate, tetrahydrofuran, N,N-dimethylformamide, and dimethyl sulfoxide.

8. An isomer, characterized in that The isomer is an isomer of the cyclic α-hydroxyimide compound according to any one of claims 1 to 4; preferably, the isomer includes stereoisomers, tautomers, and cis-trans isomers.

9. An isotope derivative, characterized in that: The isotope derivative is an isotope derivative obtained by replacing the hydrogen atoms in the cyclic α-hydroxyimide compound according to any one of claims 1 to 4 with 1 to 6 deuterium atoms or replacing the carbon atoms with 1 to 3 carbon 14 atoms.

10. A prodrug, characterized in that The prodrug is a chemical derivative of the cyclic α-hydroxyimide compound according to any one of claims 1 to 4.

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