Alpha-hydroxy ketone nitrogen-containing fused ring compound and application thereof

Abstract

The invention relates to the technical field of biological medicine, and particularly discloses an alpha-hydroxy ketone nitrogen-containing fused ring compound and application thereof. The structural general formula of the compound is as shown in formula (1), and in the formula (1), R1 is hydrogen; deuterium; a hydroxyl group; an amino group; a C1-6 alkoxy group; a C1-6 alkyl amino group; a hydroxyl group substituted by aryl or heteroaryl, an amino group, a C1-6 alkoxy group or a C1-6 alkylamino group; r2 is hydrogen; deuterium; a C1-6 alkyl group; c3-7 cycloalkyl groups; a C3-7 heterocyclic alkyl group; a C1-6 alkoxy group; a C1-6 alkyl amino group; any one of an optionally substituted aryl group or an optionally substituted heteroaryl group; x1 is any one of NH and N; x2 is any one of CH2 and CH; a single bond or a double bond; when it is a single bond, R2 is substituted only on X1; and Y is any one of CH2 and NH. 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 an α-hydroxyketone nitrogen-containing fused ring 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 interaction between HBV infection and the host immune response and environment can lead to persistent HBV infection and cause varying degrees of liver damage. The molecular mechanism of persistent HBV infection is very complex, involving HBV's unique replication mechanism, host immune system vulnerabilities and other factors. At present, it is generally believed that HBV is endocytosed through the sodium taurocholate cotransporter (sodium taurocholate cotransporting polypeptide, NTCP). Inside the cell, rcDNA (double-stranded relaxed circular DNA molecule) enters the 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) binds to HBV Pol (polymerase protein) in the cytosol to induce the packaging process, followed by the synthesis of HBV DNA. New HBV virus particles are assembled and translocated in the endoplasmic reticulum and secreted outside the hepatocyte as virus particles, thereby achieving 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 an α-hydroxyketone nitrogen-containing fused ring compound and application thereof.

[0008] Human flap endonuclease 1 (FEN-1) is a metallo-nuclease catalyzed by magnesium ions, which is mainly involved in the maturation of Okazaki fragments and the excision of 5'-flap during DNA replication. This process is substrate-specific and only recognizes the specific 5'-flap structure of the substrate DNA, regardless of the sequence of the substrate DNA. Several studies have shown that FEN-1 plays a vital role in the repair of HBV cccDNA.

[0009] The α-hydroxyketone nitrogen-containing fused ring compound provided in the present application has strong FEN-1 inhibitory activity, further inhibiting 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.

[0010] In a first aspect, the present application provides an α-hydroxyketone nitrogen-containing fused ring compound, which adopts the following technical solution:

[0011] An α-hydroxy ketone nitrogen-containing fused ring compound, the general structural formula of the α-hydroxy ketone nitrogen-containing fused ring compound is shown in formula (1):

[0012]

[0013] In the formula (1), R1 is hydrogen; deuterium; hydroxyl; amino; C 1-6 Alkoxy; C 1-6 Alkylamino; hydroxy, amino, C substituted by aryl or heteroaryl 1-6 Alkoxy or C 1-6 Any of alkylamino;

[0014] R2 is hydrogen; deuterium; C 1-6 Alkyl; C 3-7 Cycloalkyl; C 3-7 Heterocycloalkyl; C 1-6 Alkoxy; C 1-6 alkylamino; any of optionally substituted aryl or optionally substituted heteroaryl;

[0015] X1 is any of NH and N;

[0016] X2 is any of CH2 and CH;

[0017] Y is any of CH2 and NH;

[0018] is a single bond or a double bond; when When it is a single bond, R2 is substituted only on X1.

[0019] Preferably, the structural formula of the α-hydroxyketone nitrogen-containing fused ring compound is shown in Formula (2) or Formula (3):

[0020]

[0021] In the formula (2) and formula (3), R2 is hydrogen; deuterium; C 1-6 Alkyl; C 3-7 Cycloalkyl; C 3-7 Heterocycloalkyl; C 1-6 Alkoxy; C 1-6 alkylamino; any of optionally substituted aryl or optionally substituted heteroaryl; R3 is hydrogen; deuterium; C 1-6 Alkyl; C 3-7 Cycloalkyl; C 3-7 Heterocycloalkyl; C 1-6 Alkoxy; C 1-6 Alkylamino; hydroxy, amino, C substituted by aryl or heteroaryl 1-6 Alkoxy or C 1-6 alkylamino; any of optionally substituted aryl or optionally substituted heteroaryl;

[0022] Z is any one of NH, CH2, O, and S;

[0023] is a single bond or a double bond; when When it is a single bond, R2 is substituted only on X1.

[0024] Preferably, in the formula (2) and formula (3), R2 is C substituted by an aryl or heteroaryl group. 1-6 alkyl;

[0025] R3 is optionally substituted aryl or optionally substituted heteroaryl;

[0026] X1 is N; X2 is CH; there is a carbon-nitrogen double bond between X1 and X2;

[0027] Y is any of CH2 and NH;

[0028] Z is any of NH and CH2;

[0029] is a single bond or a double bond; when When it is a single bond, R2 is substituted only on X1.

[0030] Preferably, the structural formula of the α-hydroxyketone nitrogen-containing fused ring compound is any one of formula (4) to formula (13).

[0031] The structural formulas of formula (4) to formula (13) are as follows:

[0032]

[0033]

[0034] Formula (4) - 9-hydroxy-2,3,4,8-tetrahydro-1H-pyrido[1,2-a]pyrazine-1,8-dione.

[0035] Formula (5) - 9-hydroxy-2H-pyrido[1,2-a]pyrazine-1,8-dione.

[0036] Formula (6)—5-Hydroxy-2,3,4,6-tetrahydro-1H-pyrido[2,1-f][1,2,4]triazine-4,6-dione.

[0037] Formula (7)—5-Hydroxy-4,6-dihydro-1H-pyrido[2,1-f][1,2,4]triazine-4,6-dione.

[0038] Formula (8) - 5-Hydroxy-7-{[(2-hydroxy-3-phenylphenyl)methyl]amino}-4,6-dihydro-3H-pyrido[2,1-f][1,2,4]triazine-4,6-dione.

[0039] Formula (9) - 5-Hydroxy-7-{[(2-hydroxy-3-phenylphenyl)methyl]amino}-4,6-dihydro-3H-pyrido[2,1-f][1,2,4]triazine-4,6-dione.

[0040] Formula (10)—5,7-dihydroxy-3-methyl-4,6-dihydro-3H-pyrido[2,1-f][1,2,4]triazine-4,6-dione.

[0041] Formula (11) - 5,7-dihydroxy-3-{[5-(7H-pyrrolo[2,3-d]pyrimidin-5-yl)-1H-pyrrol-3-yl]methyl}-4,6-dihydro-3H-pyrido[2,1-f][1,2,4]triazine-4,6-dione.

[0042] Formula (12) - 5,7-dihydroxy-3-{[5-(7H-pyrrolo[2,3-d]pyrimidin-5-yl)-1H-pyrrol-3-yl]methyl}-4,6-dihydro-3H-pyrido[2,1-f][1,2,4]triazine-4,6-dione.

[0043] Formula (13) - 5,7-dihydroxy-3-({5-[3-hydroxy-4-(hydroxymethyl)phenyl]thiophen-3-yl}methyl)-4,6-dihydro-3H-pyrido[2,1-f][1,2,4]triazine-4,6-dione.

[0044] In addition, the α-hydroxy ketone nitrogen-containing fused ring compound may also be 3-{[4-(3,4-dihydroxyphenyl)thiophen-2-yl]methyl}-5,7-dihydroxy-4,6-dihydro-3H-pyrido[2,1-f][1,2,4]triazine-4,6-dione.

[0045] In a second aspect, the present application provides the use of the above-mentioned α-hydroxyketone nitrogen-containing fused ring compound in the preparation of a drug for preventing and treating hepatitis B virus infection.

[0046] In a third aspect, the present application provides a composition, which adopts the following technical solution:

[0047] A composition comprises the above-mentioned α-hydroxy ketone nitrogen-containing fused ring compound.

[0048] Preferably, the composition is a preparation; the preparation further comprises a pharmaceutically acceptable salt.

[0049] Preferably, the pharmaceutically acceptable salts include acid addition salts and base addition salts.

[0050] In a fourth aspect, the present application also provides a solvate, which adopts the following technical solution:

[0051] A solvate, which is an association formed by the above-mentioned α-hydroxyketone nitrogen-containing fused ring compound and one or more solvent molecules.

[0052] Preferably, the solvent includes any one or more of water, methanol, ethanol, isopropanol, ethyl acetate, tetrahydrofuran, N,N-dimethylformamide, and dimethyl sulfoxide.

[0053] Preferably, the solvate is a hydrate.

[0054] In summary, this application has the following beneficial effects:

[0055] The α-hydroxyketone nitrogen-containing fused ring 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. It is expected to be developed into a new anti-hepatitis B drug to fill the gap in the treatment mechanism of anti-hepatitis B drugs. BRIEF DESCRIPTION OF THE DRAWINGS

[0056] Figure 1 It is a synthetic route diagram of a specific compound represented by formula (2).

[0057] Figure 2 It is a synthetic route diagram of a specific compound represented by formula (3). DETAILED DESCRIPTION

[0058] Explanation of terms

[0059] 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.

[0060] 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.

[0061] Preferably, "C 1-6 The "alkyl" is any one of methyl, ethyl, n-propyl, isopropyl and tert-butyl.

[0062] 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.

[0063] Preferably, "C 3-7 The "cycloalkyl" is any of cyclopropyl, cyclopentyl and cyclohexyl.

[0064] The term "C 3-7The 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.

[0065] The term "C 1-6 "Alkoxy" alone or in combination means a group C 1-6 Alkyl-O-, where "C 1-6 "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.

[0066] 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.

[0067] Preferably, "C 1-6 The "alkylamino" is any one of methylamino, ethylamino, isopropylamino and tert-butylamino.

[0068] 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.

[0069] 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.

[0070] The term "hydrate" refers to an association formed between water and a compound in the present application.

[0071] 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, etc. Preferably, the solvate is a hydrate.

[0072] 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.

[0073] The term "prodrug" refers to a chemical derivative of the compound of the present application, which is converted into the compound of the present application through a chemical reaction in vivo.

[0074] The term "isotopic derivative" refers to an isotopic derivative in which the hydrogen atoms in the compounds of the present application are replaced by 1 to 6 deuterium atoms, and an isotopic derivative in which the carbon atoms in the compounds of the present application are replaced by 1 to 3 carbon 14 atoms.

[0075] The present application provides an α-hydroxy ketone nitrogen-containing fused ring compound, the general structural formula of the α-hydroxy ketone nitrogen-containing fused ring compound is shown in formula (1):

[0076]

[0077] In the formula (1), R1 is hydrogen; deuterium; hydroxyl; amino; C 1-6 Alkoxy; C 1-6 Alkylamino; hydroxy, amino, C substituted by aryl or heteroaryl 1-6 Alkoxy or C 1-6 Any of alkylamino;

[0078] R2 is hydrogen; deuterium; C 1-6 Alkyl; C 3-7 Cycloalkyl; C 3-7 Heterocycloalkyl; C 1-6 Alkoxy; C 1-6 alkylamino; any of optionally substituted aryl or optionally substituted heteroaryl;

[0079] X1 is any of NH and N;

[0080] X2 is any of CH2 and CH;

[0081] Y is any of CH2 and NH;

[0082] is a single bond or a double bond; when When it is a single bond, R2 is substituted only on X1.

[0083] Preferably, the structural formula of the α-hydroxyketone nitrogen-containing fused ring compound is formula (2) or formula (3):

[0084]

[0085] In the formula (2) and formula (3), R2 is hydrogen; deuterium; C 1-6 Alkyl; C 3-7 Cycloalkyl; C 3-7 Heterocycloalkyl; C 1-6 Alkoxy; C 1-6 alkylamino; any of optionally substituted aryl or optionally substituted heteroaryl; R3 is hydrogen; deuterium; C 1-6 Alkyl; C 3-7 Cycloalkyl; C 3-7 Heterocycloalkyl; C1-6 Alkoxy; C 1-6 Alkylamino; hydroxy, amino, C substituted by aryl or heteroaryl 1-6 Alkoxy or C 1-6 alkylamino; any of optionally substituted aryl or optionally substituted heteroaryl;

[0086] Z is any one of NH, CH2, O, and S;

[0087] Y is any of CH2 and NH;

[0088] is a single bond or a double bond; when When it is a single bond, R2 is substituted only on X1.

[0089] The present application also provides a synthesis route of the above-mentioned α-hydroxy ketone nitrogen-containing fused ring compound. The following describes the synthesis route of the α-hydroxy ketone nitrogen-containing fused ring compound provided by the present application to further describe the technical solution of the present application.

[0090] like Figure 1 The product 9 in the synthetic route shown (i.e., the synthetic route of general formula 1) is a specific compound of the above formula (2).

[0091] When X1 in formula (2) is NH, X2 is CH2, Y is NH, and Z is NH, formula (2) is Figure 1 The product 9 in the synthetic route shown in .

[0092] refer to Figure 1 The synthetic route of product 9 specifically comprises the following steps:

[0093] (1) Compound 1 reacts with Boc-hydrazine under acidic conditions to obtain product 2;

[0094] (2) Product 2 is de-Bocated under TFA conditions to obtain product 3;

[0095] (3) Product 3 is cyclized with formamidine to obtain product 4;

[0096] (4) Product 4 is hydrolyzed under alkaline conditions to obtain product 5;

[0097] (5) Product 5 reacts with DPPA under alkaline conditions to obtain product 6;

[0098] (6) Product 6 is de-Bocated under TFA conditions to obtain product 7;

[0099] (7) Product 7 reacts with bromide or substituted aldehyde (ketone) to obtain product 8;

[0100] (8) Product 8 is debenzylated under palladium-carbon catalysis to obtain product 9.

[0101] like Figure 2 The product 18 in the synthetic route shown (i.e., the synthetic route of general formula 2) is a specific compound of the above formula (3).

[0102] When X1 in formula (3) is N, X2 is CH2, and Y is NH, formula (3) is Figure 2 The product 18 in the synthetic route shown in FIG.

[0103] refer to Figure 2 The synthetic route of product 18 specifically includes the following steps:

[0104] (1) Compound 10 is brominated to obtain product 11;

[0105] (2) Product 11 reacts with Pin2B2 to obtain product 12;

[0106] (3) product 12 reacts with hydrogen peroxide to obtain product 13;

[0107] (4) Product 13 is substituted with benzyl bromide to obtain product 14;

[0108] (5) Product 14 reacts with Boc-hydrazine under acidic conditions to obtain product 15;

[0109] (6) Product 15 is cyclized with formamide to obtain product 16;

[0110] (7) Product 16 is coupled with a substituted boronic acid to give product 17;

[0111] (8) Product 17 is debenzylated under palladium-carbon catalysis to give product 18.

[0112] The abbreviations of materials commonly involved in this application are shown in Table 1.

[0113] Table 1 Abbreviations of materials commonly involved in this application

[0114] Serial number abbreviation Chinese name Serial number abbreviation Chinese name 1 <![CDATA[CDCl3]]> Deuterated chloroform 5 MeOH Methanol 2 DMSO-d6 Deuterated dimethyl sulfoxide 6 TFA Trifluoroacetic acid 3 DMF N,N-Dimethylformamide 7 THF Tetrahydrofuran 4 EA Ethyl acetate 8 DCM Dichloromethane

[0115] 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.

[0116] Example

[0117] The following describes the general experimental conditions in the examples of this application.

[0118] First, the reactions in the examples of the present application were all carried out under nitrogen protection.

[0119] 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).

[0120] 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.

[0121] Table 2 Gradient elution conditions

[0122]

[0123]

[0124] 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).

[0125] The names, structural formulas and preparation processes of the α-hydroxyketone nitrogen-containing fused ring compounds provided in Examples 1-5 are specifically shown in Tables 3 and 4.

[0126] Table 3 Names and structural formulas of intermediates in each step

[0127]

[0128] Table 4 Name, structural formula and preparation process of α-hydroxyketone nitrogen-containing fused ring compounds provided in the examples

[0129]

[0130]

[0131] Example 1

[0132] This embodiment provides 9-hydroxy-2,3,4,8-tetrahydro-1H-pyrido[1,2-a]pyrazine-1,8-dione, the structural formula of which is shown in formula (4), and the specific preparation method is as follows, as shown in Tables 3 and 4.

[0133] (1) Preparation of 3-benzyloxy-4-oxo-4H-pyran-2-carboxylic acid methyl ester (1A), the structural formula of which is shown in formula (14).

[0134] 3-Benzyloxy-4-oxo-4H-pyran-2-carboxylic acid was dissolved in DMF, anhydrous K2CO3 was added, and then iodomethane was added, and the reaction was carried out overnight at 40°C. After the reaction was completed, water was added to quench, and EA (40 mL×3) was used for extraction. The organic phase was washed with saturated brine, dried and concentrated, and the crude product was purified by silica gel column chromatography to obtain a white solid, 3-benzyloxy-4-oxo-4H-pyran-2-carboxylic acid methyl ester (1A).

[0135] (2) Preparation of 3-benzyloxy-4-oxo-1,4-dihydropyridine-1-[2-((tert-butyloxycarbonyl)amino)ethyl]-2-carboxylic acid methyl ester (1B), the structural formula of which is shown in formula (15).

[0136] 3-Benzyloxy-4-oxo-4H-pyran-2-carboxylic acid methyl ester (1A) was dissolved in toluene, and then N-tert-butyloxycarbonyl-1,2-ethylenediamine and glacial acetic acid were added under nitrogen protection, and the temperature was raised to 110°C and reacted for 16 hours. The reaction solution was concentrated under reduced pressure to remove most of the toluene, and the crude product was purified by silica gel column chromatography to obtain a white solid - 3-benzyloxy-4-oxo-1,4-dihydropyridine-1-[2-((tert-butyloxycarbonyl)amino)ethyl]-2-carboxylic acid methyl ester (1B).

[0137] (3) Prepare 9-benzyloxy-3,4-dihydropyrido[1,2-a]pyrazine-1,8-dione (1C), the structural formula of which is shown in formula (16).

[0138] 3-Benzyloxy-4-oxo-1,4-dihydropyridine-1-[2-((tert-butyloxycarbonyl)amino)ethyl]-2-carboxylic acid methyl ester (1B) was dissolved in ethyl acetate (50 mL) of hydrochloric acid. After the reaction of the raw materials was completed, a saturated sodium bicarbonate solution was added to the mixture. After the reaction was completed, it was extracted with dichloromethane (40 mL) and the organic phases were combined. The organic phase was dried over anhydrous sodium sulfate and concentrated. The crude product was purified by silica gel column chromatography to obtain a light yellow oily substance, 9-benzyloxy-3,4-dihydropyrido[1,2-a]pyrazine-1,8-dione (1C).

[0139] (4) Prepare 9-hydroxy-2,3,4,8-tetrahydro-1H-pyrido[1,2-a]pyrazine-1,8-dione, the structural formula of which is shown in formula (4).

[0140] 9-Benzyloxy-3,4-dihydropyrido[1,2-a]pyrazine-1,8-dione (1C) was dissolved in DCM, and boron tribromide was added to react at room temperature for 1 hour. After the reaction was completed, the mixture was quenched with water, extracted with EA three times, and the organic phase was concentrated and purified by reverse phase high performance liquid chromatography (Pre-HPLC) to obtain a white solid, 9-hydroxy-2,3,4,8-tetrahydro-1H-pyrido[1,2-a]pyrazine-1,8-dione.

[0141] Example 2

[0142] This embodiment provides 9-hydroxy-2H-pyrido[1,2-a]pyrazine-1,8-dione, whose structural formula is shown in formula (5). The specific preparation method is as follows, see Table 3 and Table 4 for details.

[0143] (1) Prepare 3-benzyloxy-N-(4-methoxybenzyl)-4-oxo-4H-pyran-2-carboxamide (2A), the structural formula of which is shown in formula (17).

[0144] 3-Benzyloxy-4-oxo-4H-pyran-2-carboxylic acid was dissolved in DCM, 1-hydroxybenzotriazole (HOBt), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI) were added, and then 4-methoxybenzylamine was added, and the reaction was carried out at room temperature for 16 hours. After the reaction was completed, the mixture was quenched with water and extracted with dichloromethane (40 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate and concentrated, and the crude product was purified by silica gel column chromatography to obtain a white solid, 3-benzyloxy-N-(4-methoxybenzyl)-4-oxo-4H-pyran-2-carboxamide (2A).

[0145] (2) Prepare 3-benzyloxy-1-(2,2-dimethoxyethyl)-N-(4-methoxybenzyl)-4-oxo-1,4-dihydropyridine-2-carboxamide (2B), the structural formula of which is shown in formula (18).

[0146] 3-Benzyloxy-N-(4-methoxybenzyl)-4-oxo-4H-pyran-2-carboxamide (2A) was dissolved in MeOH, 2,2-dimethoxyethane-1-amine was added, and the temperature was raised to 70°C for 16 hours. After the reaction, the reaction solution was concentrated, and the crude product was purified by silica gel column chromatography to obtain a yellow solid - 3-benzyloxy-1-(2,2-dimethoxyethyl)-N-(4-methoxybenzyl)-4-oxo-1,4-dihydropyridine-2-carboxamide (2B).

[0147] (3) Prepare 9-hydroxy-2-(4-methoxybenzyl)-2H-pyrido[1,2-a]pyrazine-1,8-dione (2C), the structural formula of which is shown in formula (19).

[0148] 3-Benzyloxy-1-(2,2-dimethoxyethyl)-N-(4-methoxybenzyl)-4-oxo-1,4-dihydropyridine-2-carboxamide (2B) was dissolved in toluene, and L-camphorsulfonic acid was added. The temperature was raised to 110°C and the reaction was continued for 20 hours. After the reaction, the reaction solution was concentrated, and the crude product was purified by reverse phase silica gel column chromatography to obtain a light yellow oily substance, 9-hydroxy-2-(4-methoxybenzyl)-2H-pyrido[1,2-a]pyrazine-1,8-dione (2C).

[0149] (4) Prepare 9-hydroxy-2H-pyrido[1,2-a]pyrazine-1,8-dione, the structural formula of which is shown in formula (5).

[0150] 9-Hydroxy-2-(4-methoxybenzyl)-2H-pyrido[1,2-a]pyrazine-1,8-dione (2C) was dissolved in a mixed solvent of TFA and anisole, and the temperature was raised to 140°C for 48 hours. After the reaction, the reaction solution was concentrated and purified by Pre-HPLC to obtain a white solid, 9-hydroxy-2H-pyrido[1,2-a]pyrazine-1,8-dione.

[0151] Example 3

[0152] This embodiment provides 5-hydroxy-2,3,4,6-tetrahydro-1H-pyrido[2,1-f][1,2,4]triazine-4,6-dione. Its structural formula is shown in formula (6). The specific preparation method is as follows, see Table 3 and Table 4 for details.

[0153] (1) Prepare 3-benzyloxy-4-oxo-1,4-dihydropyridine-2-carboxylic acid (3A), the structural formula of which is shown in formula (20).

[0154] 3-Benzyloxy-4-oxo-4H-pyran-2-carboxylic acid was dissolved in ammonia water and reacted at room temperature for 16 hours. After the reaction, the mixture was concentrated and the crude product was purified by silica gel column chromatography to obtain a white solid, 3-benzyloxy-4-oxo-1,4-dihydropyridine-2-carboxylic acid (3A).

[0155] (2) Prepare 3-benzyloxy-4-oxo-1,4-dihydropyridine-2-carboxamide (3B), the structural formula of which is shown in formula (21).

[0156] 3-Benzyloxy-4-oxo-1,4-dihydropyridine-2-carboxylic acid (3A) was dissolved in DMF, and N,N-diisopropylethylamine (DIEA), HOBt, EDCI, and ammonium chloride were added and reacted at room temperature for 3 hours. The reaction solution was purified by reverse phase silica gel column chromatography to obtain a white solid, 3-benzyloxy-4-oxo-1,4-dihydropyridine-2-carboxamide (3B).

[0157] (3) Prepare 1-amino-3-benzyloxy-4-oxo-1,4-dihydropyridine-2-carboxamide (3C), the structural formula of which is shown in formula (22).

[0158] 3-Benzyloxy-4-oxo-1,4-dihydropyridine-2-carboxamide (3B) and O-(2,4-dinitrophenyl)hydroxylamine were dissolved in acetonitrile, potassium carbonate was added, and then the mixture was reacted at room temperature for 16 hours. After the reaction, the reaction solution was concentrated and the crude product was purified by silica gel column chromatography to obtain a yellow solid, 1-amino-3-benzyloxy-4-oxo-1,4-dihydropyridine-2-carboxamide (3C).

[0159] (4) Prepare 5-benzyloxy-2,3-dihydro-1H-pyrido[2,1-f][1,2,4]triazine-4,6-dione (3D), the structural formula of which is shown in formula (23).

[0160] 1-Amino-3-benzyloxy-4-oxo-1,4-dihydropyridine-2-carboxamide (3C) was dissolved in DMF and acetic acid, and paraformaldehyde was added. The temperature was then raised to 100°C for 4 hours. After the reaction, water was added to precipitate the solid, which was slurried with water and dried to obtain a yellow solid - 5-benzyloxy-2,3-dihydro-1H-pyrido[2,1-f][1,2,4]triazine-4,6-dione (3D).

[0161] (5) Prepare 5-hydroxy-2,3,4,6-tetrahydro-1H-pyrido[2,1-f][1,2,4]triazine-4,6-dione, the structural formula of which is shown in formula (6).

[0162] 5-Benzyloxy-2,3-dihydro-1H-pyrido[2,1-f][1,2,4]triazine-4,6-dione (3D) was dissolved in DCM, and boron tribromide was added. The mixture was reacted at room temperature for 2 h. After the reaction, the mixture was concentrated and the crude product was purified by Pre-HPLC to obtain a white solid, 5-hydroxy-2,3,4,6-tetrahydro-1H-pyrido[2,1-f][1,2,4]triazine-4,6-dione.

[0163] Example 4

[0164] This embodiment provides 5-hydroxy-4,6-dihydro-1H-pyrido[2,1-f][1,2,4]triazine-4,6-dione, whose structural formula is shown in formula (7). The specific preparation method is as follows, see Table 3 and Table 4 for details.

[0165] (1) Prepare 5-benzyloxy-2-methoxy-2,3-dihydro-1H-pyrido[2,1-f][1,2,4]triazine-4,6-dione (4A), the structural formula of which is shown in formula (24).

[0166] 1-Amino-3-benzyloxy-4-oxo-1,4-dihydropyridine-2-carboxamide (3C) was dissolved in triethyl orthoformate, p-toluenesulfonic acid was added, and then the temperature was raised to 100°C for reaction for 4 hours. After the reaction was completed, the solid was precipitated by cooling, and filtered to obtain a yellow solid - 5-benzyloxy-2-methoxy-2,3-dihydro-1H-pyrido[2,1-f][1,2,4]triazine-4,6-dione (4A).

[0167] (2) Prepare 5-benzyloxy-1H-pyrido[2,1-f][1,2,4]triazine-4,6-dione (4B), the structural formula of which is shown in formula (25).

[0168] 5-Benzyloxy-2-methoxy-2,3-dihydro-1H-pyrido[2,1-f][1,2,4]triazine-4,6-dione (4A) was dissolved in THF, LiHMDS was added, and the reaction was carried out at room temperature for 1 hour. After the reaction was completed, 5 mL of water was added for dilution, and 10 mL of EA was added for extraction. The organic phase was collected, dried and concentrated, and the crude product was purified by silica gel column chromatography to obtain a white solid, 5-benzyloxy-1H-pyrido[2,1-f][1,2,4]triazine-4,6-dione (4B).

[0169] (3) Prepare 5-hydroxy-4,6-dihydro-1H-pyrido[2,1-f][1,2,4]triazine-4,6-dione, the structural formula of which is shown in formula (7).

[0170] 5-Benzyloxy-1H-pyrido[2,1-f][1,2,4]triazine-4,6-dione (4B) was dissolved in DCM, and boron tribromide was added. The mixture was reacted at room temperature for 2 h. After the reaction, the mixture was concentrated and the crude product was purified by Pre-HPLC to obtain a white solid, 5-hydroxy-4,6-dihydro-1H-pyrido[2,1-f][1,2,4]triazine-4,6-dione.

[0171] Example 5

[0172] This embodiment provides 5-hydroxy-7-{[(2-hydroxy-3-phenylphenyl)methyl]amino}-4,6-dihydro-3H-pyrido[2,1-f][1,2,4]triazine-4,6-dione. Its structural formula is shown in formula (8), and its preparation method is as follows, as shown in Table 3.

[0173] Using 2-hydroxy-[1,1'-biphenyl]-3-carboxaldehyde as raw material (220 mg, 1.13 mmol), a yellow solid, 5-hydroxy-7-{[(2-hydroxy-3-phenylphenyl)methyl]amino}-4,6-dihydro-3H-pyrido[2,1-f][1,2,4]triazine-4,6-dione (9.5 mg, yield: 12%), was prepared according to the synthetic route of general formula 1.

[0174] The test results are as follows:

[0175] MS (ESI) m / z = 377.2 [M + H] + .

[0176] 1 H NMR (DMSO-d6, 400MHz): δ12.25 (s, 1H), 8.75 (s, 1H), 7.84 (s, 1H), 7.51-7.49 (m, 2H), 7.45-7.40 (m, 3H), 7.3 4-7.30 (m, 1H), 7.20-7.18 (m, 1H), 7.13-7.11 (m, 1H), 6.91-6.87 (m, 1H), 6.16 (s, 1H), 2.63 (d, J=3.6Hz, 2H).

[0177] In addition, the present application also provides Examples 6-10, and the names and structural formulas of the corresponding compounds are shown in Table 5. Those skilled in the art can prepare the following compounds by the preparation methods given in the present application and the preparation methods available in the art.

[0178] Table 5 Name and structural formula of α-hydroxyketone nitrogen-containing fused ring compounds provided in the examples (II)

[0179]

[0180] The results of biological tests on the above-mentioned α-hydroxyketone nitrogen-containing fused ring compounds are described in detail below to further illustrate the technical solution of the present application.

[0181] Test 1

[0182] The test subjects are the α-hydroxyketone nitrogen-containing fused ring compounds provided in Examples 1-10 of the present application, and the test type is to detect the inhibitory ability of the above compounds on FEN-1.

[0183] The details are as follows:

[0184] (1) Test method

[0185] A. Expression and purification of wild-type human FEN-1 protein

[0186] 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).

[0187] B. Construction of FEN-1 enzyme activity detection system based on fluorescence signal

[0188] 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.

[0189] Table 6 Construction of DNA substrate containing HBV sequence

[0190]

[0191] The annealing program was as follows: 95°C, 5 min; then cooled to 25°C.

[0192] 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 "flap". Different concentration gradients of the test compound (compounds provided in Examples 1-10) 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:

[0193] Inhibition rate and IC 50 The calculation formula is as follows:

[0194]

[0195] IC 50of the compound is calculated by XL fit software-Fit models(205).

[0196] (2) Test results

[0197] The test results are shown in Table 7. 50 (half inhibitory concentration) indicates the concentration of the test compound when it inhibits half the amount of FEN-1 protein; the stronger the inhibitory ability, the lower the concentration.

[0198] Table 7 Activity data of α-hydroxyketone nitrogen-containing fused ring compounds provided in Examples 1-10 for inhibiting FEN-1

[0199]

[0200]

[0201] It can be seen from Table 7 that the α-hydroxyketone nitrogen-containing fused ring compound provided in the present application can effectively inhibit the activity of FEN-1 protein.

[0202] 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. An α-hydroxyketone nitrogen-containing fused ring compound, characterized in that: The general structural formula of the α-hydroxyketone nitrogen-containing fused ring compound is shown in formula (1): In the formula (1), R1 is hydrogen; deuterium; hydroxyl; amino; C 1-6 Alkoxy; C 1-6 Alkylamino; hydroxy, amino, C substituted by aryl or heteroaryl 1-6 Alkoxy or C 1-6 Any of alkylamino; R2 is hydrogen; deuterium; C 1-6 Alkyl; C 3-7 Cycloalkyl; C 3-7 Heterocycloalkyl; C 1-6 Alkoxy; C 1-6 alkylamino; any of optionally substituted aryl or optionally substituted heteroaryl; X1 is any of NH and N; X2 is any of CH2 and CH; Y is any of CH2 and NH; is a single bond or a double bond; when When it is a single bond, R2 is substituted only on X1.

2. The α-hydroxyketone nitrogen-containing fused ring compound according to claim 1, characterized in that: The structural formula of the α-hydroxyketone nitrogen-containing fused ring compound is formula (2) or formula (3): In the formula (2) and formula (3), R2 is hydrogen; deuterium; C 1-6 Alkyl; C 3-7 Cycloalkyl; C 3-7 Heterocycloalkyl; C 1-6 Alkoxy; C 1-6 alkylamino; any of optionally substituted aryl or optionally substituted heteroaryl; R3 is hydrogen; deuterium; C 1-6 Alkyl; C 3-7 Cycloalkyl; C 3-7 Heterocycloalkyl; C 1-6 Alkoxy; C 1-6 Alkylamino; hydroxy, amino, C substituted by aryl or heteroaryl 1-6 Alkoxy or C 1-6 alkylamino; any of optionally substituted aryl or optionally substituted heteroaryl; Z is any one of NH, CH2, O, and S; Y is any of CH2 and NH; is a single bond or a double bond; when When it is a single bond, R2 is substituted only on X1.

3. The α-hydroxyketone nitrogen-containing fused ring compound according to claim 2, characterized in that: In the formula (2) and formula (3), R2 is a C substituted by an aryl or heteroaryl group. 1-6 alkyl; R3 is optionally substituted aryl or optionally substituted heteroaryl; X1 is N; X2 is CH; there is a carbon-nitrogen double bond between X1 and X2; Y is any of CH2 and NH; Z is either NH or CH2.

4. The α-hydroxyketone nitrogen-containing fused ring compound according to claim 1, characterized in that: The structural formula of the α-hydroxyketone nitrogen-containing fused ring compound is any one of formula (4) to formula (13):

5. Use of the α-hydroxyketone nitrogen-containing fused ring 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 α-hydroxyketone nitrogen-containing fused ring 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 α-hydroxyketone nitrogen-containing fused ring 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; and the solvate is a hydrate.

8. An isomer, characterized in that The isomer is an isomer of the α-hydroxyketone nitrogen-containing fused ring 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 isotopic derivative is an isotopic derivative obtained by replacing the hydrogen atoms in the α-hydroxyketone nitrogen-containing fused ring 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 α-hydroxyketone nitrogen-containing fused ring compound according to any one of claims 1 to 4.

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