A compound based on the structure of 6-methylfurano[2,3-d]pyrimidine-4(3H)-one, its preparation method and application

Abstract

This invention belongs to the field of pharmaceutical synthesis technology, specifically relating to a compound based on the 6-methylfurano[2,3-d]pyrimidine-4(3H)-one structure, its preparation method, and its applications. The compound provided by this invention, based on the 6-methylfurano[2,3-d]pyrimidine-4(3H)-one structure, enhances the chelation effect between the compound and zinc ions in histone deacetylases by combining the pharmacophore of a DNA methyltransferase 3A inhibitor with an isohydroxamic acid fragment, thus exhibiting strong HDAC inhibitory activity while maintaining DNMT3A inhibition. The compound provided by this invention shows significantly higher inhibitory activity against human non-small cell lung cancer cells H460 than against H460 DNMT3A-HDAC6 double knockout cells, demonstrating in vivo activity against human non-small cell lung cancer and possessing application value as a therapeutic agent for diseases and disorders related to DNMT3A and HDAC6.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical synthesis technology, specifically relating to a compound based on the structure of 6-methylfurano[2,3-d]pyrimidine-4(3H)-one, its preparation method, and its application. Background Technology

[0002] Epigenetic regulation is closely related to tumorigenesis, progression, and drug resistance. Currently, several drugs targeting epigenetic enzymes have been approved by the FDA and used in cancer treatment. For example, the DNA methyltransferase inhibitor 5-azacytidine (5-AzaC) is used to treat myelodysplastic syndromes and acute myeloid leukemia, and histone deacetylase (HDAC) inhibitors (such as Voronostat, Belinostat, and Romidepsin) have been approved for the treatment of cutaneous and peripheral T-cell lymphomas.

[0003] In 2022, researchers discovered that the non-nucleoside DNA methyltransferase 3A (DNMT3A) inhibitor DY-46 has high activity (IC50). 50 It exhibits high selectivity (1.3 ± 0.22 μM) and can occupy both the SAM-cofactor pocket and cytosine pocket of DNMT3A. Its analogue DY-46-2 shows high IC50 for DNMT3A. 50 The concentration was 0.39±0.23 μM, showing excellent selectivity relative to DNMT1 (33.3 times), DNMT3B (269 times), and G9a (more than 1000 times). DY-46 and DY-46-2 significantly inhibited cancer cell proliferation [Zhang J, et al. Cancers (Basel), 2020, 12(8). 11-16].

[0004] In cells lacking DNMT3A, histone deacetylase 6 (HDAC6) is significantly upregulated, which may be one of the reasons why single DNMT inhibitors are mainly used to treat leukemia and are accompanied by many adverse reactions. How to develop dual-target inhibitors that selectively target both DNMT3A and HDAC6 to provide a synergistic effect for therapeutic applications remains a gap in research. Summary of the Invention

[0005] The purpose of this invention is to provide a compound based on the 6-methylfurano[2,3-d]pyrimidine-4(3H)-one structure, its preparation method and application. The compound provided by this invention has strong DMNT3A inhibition and HDAC6 inhibition.

[0006] To achieve the above objectives, the present invention provides the following technical solution:

[0007] This invention provides a compound based on the structure of 6-methylfurano[2,3-d]pyrimidine-4(3H)-one, as shown in Formula I:

[0008]

[0009] In Formula I, R is an amino group, a substituted amino group, a hydroxyl group, or an alkoxy group; L is a benzene ring, a substituted benzene ring, an aromatic heterocycle, a substituted aromatic heterocycle, or an alkyl chain; and n is 1, 2, 3, or 4.

[0010] Preferably, in the R of Formula I, the substituted amino group includes any one of the following structural groups;

[0011]

[0012] The alkoxy group includes methoxy or ethoxy.

[0013] Preferably, in formula I, the substituted benzene ring in L is:

[0014]

[0015] The aromatic heterocycle is:

[0016]

[0017] The substituted aromatic heterocycles include:

[0018]

[0019] The alkyl chain is:

[0020]

[0021] Preferably, the compound based on the 6-methylfurano[2,3-d]pyrimidine-4(3H)-one structure comprises any one of the following structural compounds:

[0022]

[0023]

[0024] The present invention also provides a method for preparing the compound based on the 6-methylfurano[2,3-d]pyrimidine-4(3H)-one structure described above, comprising the following steps:

[0025] (1) 2-chloro-3-oxobutyric acid acetic acid was mixed with malononitrile to carry out a cyclization reaction to obtain a compound with the structure shown in Formula II;

[0026] (2) The compound with the structure shown in Formula II is mixed with formic acid and acetic anhydride to carry out a cyclization reaction to obtain the compound with the structure shown in Formula III;

[0027] (3) The compound with the structure shown in Formula III is mixed with a substituted alkane, a first basic compound and a first organic solvent to carry out a substitution reaction to obtain the compound with the structure shown in Formula IV;

[0028] (4) The compound with the structure shown in Formula IV is mixed with a substituted amine, a second basic compound, and a second organic solvent to carry out a substitution reaction to obtain the compound with the structure shown in Formula V.

[0029] The substituted amine has the structural formula shown in Formula VIII:

[0030] RH type VIII;

[0031] (5) The compound with the structure shown in Formula V is mixed with a third basic compound and a third solvent and subjected to a hydrolysis reaction to obtain the compound with the structure shown in Formula VI;

[0032] (6) The compound with the structure shown in Formula VI is mixed with an amino-substituted carboxylic acid ester, a condensing agent and a fourth organic solvent to carry out a condensation reaction to obtain the compound with the structure shown in Formula VII.

[0033] (7) The compound with the structure shown in Formula VII is mixed with hydroxylamine, water, a fourth basic compound and a fifth organic solvent to carry out a hydroxylamine hydrolysis reaction to obtain the compound based on the 6-methylfurano[2,3-d]pyrimidine-4(3H)-one structure;

[0034]

[0035] Preferably, the molar ratio of the compound with the structure shown in Formula II to formic acid is 1:10 to 30; the molar ratio of the compound with the structure shown in Formula II to acetic anhydride is 1:5 to 10.

[0036] Preferably, the molar ratio of the compound with the structure shown in Formula III to the substituted alkane is 1:3 to 10; the molar ratio of the compound with the structure shown in Formula III to the first basic compound is 1:1.2 to 5; and the mass-volume ratio of the compound with the structure shown in Formula III to the first organic solvent is 1:5 to 30.

[0037] Preferably, the molar ratio of the compound with the structure shown in Formula VI to the amino-substituted carboxylic acid ester is 1:0.8 to 1.2; the molar ratio of the compound with the structure shown in Formula VI to the condensing agent is 1:1 to 2; and the mass-volume ratio of the compound with the structure shown in Formula VI to the fourth organic solvent is 1:5 to 50.

[0038] Preferably, the mass ratio of hydroxylamine to water is 1:1 to 10; the molar ratio of the compound with the structure shown in Formula VII to hydroxylamine is 1:20 to 100; the molar ratio of the compound with the structure shown in Formula VII to the fourth basic compound is 1:5 to 20; and the mass-volume ratio of the compound with the structure shown in Formula VII to the fifth organic solvent is 1:5 to 50.

[0039] The present invention also provides the use of compounds based on the 6-methylfurano[2,3-d]pyrimidine-4(3H)-one structure as described in the above-described scheme, or compounds based on the 6-methylfurano[2,3-d]pyrimidine-4(3H)-one structure obtained by the preparation method described in the above-described scheme, in the preparation of medicaments for treating diseases related to DNA methyltransferase 3A or histone deacetylase.

[0040] This invention provides a compound based on the 6-methylfurano[2,3-d]pyrimidine-4(3H)-one structure. This compound, based on the 6-methylfurano[2,3-d]pyrimidine-4(3H)-one structure, enhances the chelation of the compound with zinc ions in histone deacetylase (HDAC) by combining the pharmacophore of a DNA methyltransferase 3A (DNMT3A) inhibitor with an isohydroxamic acid fragment, thus exhibiting strong HDAC inhibitory activity while maintaining DNMT3A inhibition.

[0041] This invention also provides a method for preparing the compound based on the 6-methylfurano[2,3-d]pyrimidine-4(3H)-one structure described above. The preparation method provided by this invention is simple, convenient to operate, highly feasible, and has the potential for large-scale industrial application.

[0042] This invention also provides the application of the compounds based on the 6-methylfurano[2,3-d]pyrimidine-4(3H)-one structure described above in the preparation of drugs for treating diseases related to DNA methyltransferase 3A or histone deacetylase. The compounds provided by this invention exhibit selectivity for DNMT3A and HDAC6, and possess strong inhibitory effects on both DNMT3A and HDAC6. Their inhibitory activity against H460 human non-small cell lung cancer cells is significantly higher than that against H460 DNMT3A-HDAC6 double knockout cells. They demonstrate in vivo activity against human non-small cell lung cancer and have potential application value as a therapeutic agent for diseases and dysregulations related to DNMT3A and HDAC6. Attached Figure Description

[0043] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0044] Figure 1 Cellular thermal transfer analysis of compound FQX-1D;

[0045] Figure 2 Methylation dot blot analysis of compound FQX-1D;

[0046] Figure 3 The expression level of Ac-α-tubulin in H460 cells treated with compound FQX-1D;

[0047] Figure 4 Enzyme activity assay for compound FQX-1D. Detailed Implementation

[0048] This invention provides a compound based on the structure of 6-methylfurano[2,3-d]pyrimidine-4(3H)-one, as shown in Formula I:

[0049]

[0050] Wherein, R is an amino group, a substituted amino group, a hydroxyl group, or an alkoxy group; L is a benzene ring, a substituted benzene ring, an aromatic heterocycle, a substituted aromatic heterocycle, or an alkyl chain; and n is 1, 2, 3, or 4.

[0051] In this invention, in Formula I, the structure of the substituted amino group in R is preferably -NR1R2; R1 and R2 independently preferably include C1-C6 alkyl, substituted C1-C6 alkyl, C1-C6 alkenyl, substituted C1-C6 alkenyl, C3-C20 cycloalkyl, substituted C3-C20 cycloalkyl, 3-20 membered heterocyclic group or substituted 3-20 membered heterocyclic group, or R1 and R2 preferably together form C3-C20 cycloalkyl, substituted C3-C20 cycloalkyl, 3-20 membered heterocyclic group or substituted 3-20 membered heterocyclic group.

[0052] In this invention, the C1-C6 alkyl groups in R1 and R2 preferably include methyl, ethyl, n-propyl, isopropyl, isobutyl, sec-butyl, n-butyl, n-pentyl, or isopentyl; the C1-C6 alkenyl groups preferably include allyl; the C3-C20 cycloalkyl groups preferably have a monocyclic, polycyclic, spirocyclic, or bridged ring structure; the C3-C20 cycloalkyl groups preferably include cyclopentyl, cyclohexyl, methyl-substituted cyclohexyl, cycloheptyl, or cyclooctyl; the 3-20 membered heterocyclic groups preferably have a monocyclic, polycyclic, spirocyclic, or bridged ring structure; the heteroatoms in the 3-20 membered heterocyclic groups preferably include one or more of N, O, and S; the number of heteroatoms in the 3-20 membered heterocyclic groups is preferably 1-5, specifically 1, 2, 3, 4, or 5; the 3-20 membered heterocyclic groups preferably include pyrrolidinyl, morpholinyl, piperazine, methyl-substituted piperazine, or azaheptanyl.

[0053] In this invention, the number of substituents (substituents refer to one or more hydrogen atoms on a group being replaced by a substituent) of the substituted C1-C6 alkyl, substituted C1-C6 alkenyl, substituted C3-C20 cycloalkyl, and substituted 3-20 heterocyclic groups in R1 and R2 is preferably independently one or more, specifically one, two, or three; the number of substituents of the substituted C1-C6 alkyl, substituted C1-C6 alkenyl, substituted C3-C20 cycloalkyl, and substituted 3-20 heterocyclic groups is independently... Preferred groups include halogens, cyano groups, oxygen (i.e., two hydrogen atoms on the same carbon atom of the group are replaced by =O), C1-C6 alkyl groups, halogenated C1-C6 alkyl groups, C1-C6 alkoxy groups, C6-C10 aryl groups, C6-C10 aryloxy groups, C2-C10 ester groups, C2-C10 acyl groups, C2-C10 amide groups, carboxyl groups, nitro groups, hydroxyl groups, amino groups, amino groups substituted with one C1-C6 alkyl group, amino groups substituted with two C1-C6 alkyl groups, or mercapto groups substituted with C1-C6 alkyl groups.

[0054] In this invention, the halogen preferably includes fluorine, chlorine, or bromine; the C1-C6 alkyl preferably includes methyl, ethyl, propyl, isopropyl, isobutyl, sec-butyl, n-butyl, n-pentyl, or isopentyl; the halogenated C1-C6 alkyl preferably includes trifluoromethyl, 2-chloroethyl, or 3-chloropropyl; the C1-C6 alkoxy preferably includes methoxy, ethoxy, isopropoxy, or trifluoromethoxy; the C6-C10 aryl preferably includes phenyl; the C6-C10 aryloxy preferably includes phenoxy; the C2-C10 ester preferably includes alkyl-COO- or alkyl-OOC-; the alkyl-COO- preferably includes -CH2COO- or -CH2CH2COO-; the alkyl-OOC- preferably includes CH3COO- or CH3CH2COO-; The acyl group of C2 to C10 is preferably alkyl-CO-; the alkyl-CO- preferably includes CH3CO- or CH3CH2CO-; the amide group of C2 to C10 preferably includes alkylNHC(O)- or arylNHC(O)-; the alkylNHC(O)- preferably includes CH3NHCO-, (CH3)2NCO- or CH3CH2NHCO-; the arylNHC(O)- preferably includes PhNHCO- or PhCH3NCO-; the amino group with one C1 to C6 alkyl substituted preferably includes methylamino, ethylamino or isopropylamino; the amino group with two C1 to C6 alkyl substituted preferably includes dimethylamino or diethylamino; the mercapto group with C1 to C6 alkyl substituted (C1 to C6 alkyl-S-) preferably includes methylthio, ethylthio or isopropylthio.

[0055] In a specific embodiment of the present invention, in the R of Formula I, the substituted amino group preferably includes any one of the following structural groups;

[0056]

[0057] In this invention, the alkoxy group in R of Formula I preferably includes methoxy or ethoxy.

[0058] In this invention, in formula I, the substituted benzene ring is preferably an alkyl-substituted benzene ring; the alkyl-substituted benzene ring is preferably a C1-C10 alkyl-substituted benzene ring; the C1-C10 alkyl-substituted benzene ring is preferably a methyl-substituted benzene ring; the preferred structural formula of the methyl-substituted benzene ring is:

[0059]

[0060] In this invention, in formula I, the aromatic heterocycle in L is preferably a nitrogen-substituted aromatic heterocycle; the preferred structural formula of the nitrogen-substituted aromatic heterocycle is:

[0061]

[0062] In this invention, in formula I, the substituted aromatic heterocycle in L is preferably an alkyl-substituted aromatic heterocycle; the alkyl-substituted aromatic heterocycle is preferably a C1-C8 alkyl-substituted aromatic heterocycle, and the C1-C8 alkyl-substituted aromatic heterocycle is preferably a methyl-substituted aromatic heterocycle; the preferred structural formula of the methyl-substituted aromatic heterocycle is:

[0063]

[0064] In this invention, in formula I, the alkyl chain in L is preferably a C1-C30 alkyl chain; the C1-C30 alkyl chain is preferably a C1-C8 alkyl chain; the C1-C8 alkyl chain is preferably a hexane chain; the structural formula of the hexane chain is:

[0065]

[0066] In this invention, in Formula I, n is preferably 1, 2 or 3.

[0067] In this invention, the compound based on the 6-methylfurano[2,3-d]pyrimidin-4(3H)-one structure preferably comprises any one of the following structural compounds:

[0068]

[0069]

[0070] The present invention also provides a method for preparing the compound based on the 6-methylfurano[2,3-d]pyrimidine-4(3H)-one structure described above, comprising the following steps:

[0071] (1) 2-chloro-3-oxobutyric acid acetic acid was mixed with malononitrile to carry out a cyclization reaction to obtain a compound with the structure shown in Formula II;

[0072] (2) The compound with the structure shown in Formula II is mixed with formic acid and acetic anhydride to carry out a cyclization reaction to obtain the compound with the structure shown in Formula III;

[0073] (3) The compound with the structure shown in Formula III is mixed with a substituted alkane, a basic compound and an organic solvent to carry out a substitution reaction to obtain the compound with the structure shown in Formula IV;

[0074] (4) The compound with the structure shown in Formula IV is mixed with a substituted amine, a basic compound and an organic solvent to carry out a substitution reaction to obtain the compound with the structure shown in Formula V;

[0075] The substituted amine has the structural formula shown in Formula VIII:

[0076] RH type VIII;

[0077] (5) The compound with the structure shown in Formula V is mixed with an alkaline compound and a solvent and subjected to a hydrolysis reaction to obtain the compound with the structure shown in Formula VI;

[0078] (6) The compound with the structure shown in Formula VI is mixed with an amino-substituted carboxylic acid ester, a condensing agent and an organic solvent to carry out a condensation reaction to obtain the compound with the structure shown in Formula VII.

[0079] (7) The compound with the structure shown in Formula VII is mixed with hydroxylamine, water, a basic compound and an organic solvent to carry out a hydroxylamine hydrolysis reaction to obtain the compound based on the 6-methylfurano[2,3-d]pyrimidine-4(3H)-one structure;

[0080]

[0081] In this invention, the meanings of the substituent groups (R and L) and the degree of polymerization (n) in each structural formula are consistent and will not be repeated here.

[0082] This invention involves mixing 2-chloro-3-oxobutyric acid acetic acid with malononitrile (denoted as the first mixture) and performing a cyclization reaction (denoted as the first cyclization reaction) to obtain a compound with the structure shown in Formula II. In this invention, the molar ratio of 2-chloro-3-oxobutyric acid acetic acid to malononitrile is preferably 1–1.1:1–1.5, specifically 1:1, 1:1.2, 1:1.4, 1:1.5, 1.05:1, 1.05:1.2, 1.05:1.4, 1.05:1.5, 1.1:1, 1.1:1.1, 1.1:1.3, or 1.1:1.5.

[0083] In this invention, the temperature of the first cyclization reaction is preferably room temperature, and the reaction time is preferably 4.5 to 8 hours, specifically 4.5 hours, 5 hours, 6 hours, 6.5 hours, or 7 hours.

[0084] In this invention, the first cyclization reaction is preferably carried out in the presence of an alcohol solvent and sodium ethoxide; the alcohol solvent is preferably ethanol; the volume-to-mass ratio of the alcohol solvent to 2-chloro-3-oxobutyric acid is preferably (5-20) mL:1g, specifically 5 mL:1g, 6 mL:1g, 10 mL:1g, or 15 mL:1g; the molar ratio of sodium ethoxide to 2-chloro-3-oxobutyric acid is preferably 1-1.2:1-3, specifically 1:1, 1:2, 1:3, 1.1:3, 1.1:2, 1.1:1, 1:1.2, 1.2:1, 1.2:2, or 1.2:3.

[0085] In this invention, the first cyclization reaction preferably includes sequential concentration of the reaction solution, re-dissolution of the residue, and solid-liquid separation; the reagents used for re-dissolution preferably include petroleum ether and ethyl acetate; the volume ratio of petroleum ether to ethyl acetate is preferably 3:1; and the solid-liquid separation is preferably vacuum filtration.

[0086] In a specific embodiment of the present invention, the first mixing is preferably carried out by premixing ethyl 2-chloro-3-oxobutyrate, malononitrile, and an alcohol solvent at -5 to -20°C to obtain a premixed solution, and then adding sodium ethoxide solution dropwise to the premixed solution while stirring; the concentration of the sodium ethoxide solution is preferably 5% to 18%, specifically 5%, 8%, 10%, 15%, or 18%; the solvent of the sodium ethoxide solution preferably includes ethanol; the dropping rate of the dropwise addition while stirring is preferably 20 to 60 drops / minute, specifically 30 drops / minute or 50 drops / minute.

[0087] After obtaining the compound with the structure shown in Formula II, the present invention mixes the compound with formic acid and acetic anhydride to carry out a cyclization reaction (denoted as the second cyclization reaction) to obtain the compound with the structure shown in Formula III. In the present invention, the molar ratio of the compound with the structure shown in Formula II to formic acid is preferably 1:10 to 30, specifically 1:12, 1:15, 1:20, 1:25 or 1:28.

[0088] In this invention, the molar ratio of the compound with the structure shown in Formula II to acetic anhydride is preferably 1:5 to 10, specifically 1:6, 1:7, 1:8 or 1:9.

[0089] In this invention, the second cyclization reaction is preferably carried out under reflux conditions, and the holding time is preferably 36 to 60 hours, specifically 36 hours, 40 hours, 48 ​​hours, 52 hours or 56 hours.

[0090] In this invention, the second cyclization reaction preferably further includes sequentially concentrating the reaction solution, redissolving the residue, adjusting the pH value to 7, stirring, and solid-liquid separation; the redissolving reagent is preferably water; the pH adjusting reagent is preferably a saturated sodium carbonate aqueous solution; the stirring temperature is preferably room temperature, and the stirring time is preferably 2 hours; the solid-liquid separation is preferably vacuum filtration.

[0091] After obtaining the compound with the structure shown in Formula III, the present invention mixes the compound with a substituted alkane, a basic compound (denoted as the first basic compound), and an organic solvent (denoted as the first organic solvent) to carry out a substitution reaction (denoted as the first substitution reaction) to obtain the compound with the structure shown in Formula IV. In the present invention, the substituted alkane preferably includes dibromosubstituted alkane or bromochlorosubstituted alkane; the dibromosubstituted alkane preferably includes 1,2-dibromoethane or 1,3-dibromopropane; the bromochlorosubstituted alkane preferably includes 1-bromo-2-chloroethane or 1-bromo-3-chloropropane; the molar ratio of the compound with the structure shown in Formula III to the substituted alkane is preferably 1:3 to 10, specifically 1:4, 1:5, 1:6, 1:7, or 1:9.

[0092] In this invention, the first alkaline compound preferably includes cesium carbonate or potassium carbonate; the molar ratio of the compound with the structure shown in Formula III to the first alkaline compound is preferably 1:1.2 to 5, specifically 1:1.2, 1:1.5, 1:2, 1:3, 1:4 or 1:5.

[0093] In this invention, the first organic solvent preferably includes DMF; the mass-to-volume ratio of the compound with the structure shown in Formula III to the first organic solvent is preferably 1g:(5-30)mL, specifically 1g:7mL, 1g:10mL, 1g:15mL, 1g:18mL, 1g:21mL, 1g:25mL or 1g:28mL.

[0094] In this invention, the temperature of the first substitution reaction is preferably 20 to 50°C, specifically 20°C, 30°C, 40°C or 50°C, and the holding time is preferably 6 to 12 hours, specifically 8 hours or 10 hours.

[0095] In this invention, the process after the first substitution reaction preferably includes sequentially concentrating the reaction solution, redissolving the residue, washing the organic layer with water, washing with saturated brine, drying, solid-liquid separation, concentration, and purification; the reagent used for redissolving is preferably ethyl acetate; the number of water washings is preferably 3 or more; the number of saturated brine washings is preferably 1 or more; the drying is preferably drying with anhydrous sodium sulfate; the solid-liquid separation is preferably vacuum filtration; and the purification is preferably column chromatography purification.

[0096] After obtaining the compound with the structure shown in Formula IV, the present invention mixes the compound with a substituted amine, a basic compound (denoted as the second basic compound), and an organic solvent (denoted as the second organic solvent) to carry out a substitution reaction (denoted as the second substitution reaction) to obtain the compound with the structure shown in Formula V. In the present invention, the substituted amine preferably includes one or more of dimethylamine, diethylamine, morpholine, piperidine, pyrrolidine, N-methylpiperazine, N-ethylpiperazine, and isopropylamine; the molar ratio of the compound with the structure shown in Formula IV to the substituted amine is preferably 1:1 to 3, specifically 1:1.1, 1:1.3, 1:1.7, 1:2.1, or 1:2.7.

[0097] In this invention, the second basic compound preferably includes cesium carbonate or potassium carbonate; the molar ratio of the compound with the structure shown in Formula IV to the second basic compound is preferably 1:2 to 5, specifically 1:3 or 1:4.

[0098] In this invention, the second organic solvent preferably includes one or more of DMF, acetonitrile and tetrahydrofuran; the mass-volume ratio of the compound with the structure shown in Formula IV to the second organic solvent is preferably 1g:(10-50)mL, specifically 1g:20mL, 1g:30mL or 1g:40mL.

[0099] In this invention, the temperature of the second substitution reaction is preferably 20 to 80°C, specifically 20°C, 40°C, 60°C or 70°C, and the holding time is preferably 4 to 8 hours, specifically 5 hours or 6 hours.

[0100] In this invention, the second substitution reaction preferably further includes sequentially concentrating the reaction solution, redissolving the residue, washing with water, washing with saturated brine, drying, solid-liquid separation, and concentration; the reagent used for redissolving is preferably dichloromethane; the number of times the water is washed is preferably two or more; the number of times the saturated brine is washed is preferably one or more; the drying is preferably anhydrous sodium sulfate drying; and the solid-liquid separation is preferably vacuum filtration.

[0101] After obtaining the compound with the structure shown in Formula V, the present invention mixes the compound with the basic compound (denoted as the third basic compound) and a solvent (denoted as the third solvent) to carry out a hydrolysis reaction to obtain the compound with the structure shown in Formula VI. In the present invention, the third basic compound preferably includes one or more of sodium hydroxide and potassium hydroxide; the molar ratio of the compound with the structure shown in Formula V to the third basic compound is preferably 1:5 to 10, specifically 1:6, 1:7, 1:8 or 1:9.

[0102] In this invention, the third solvent preferably includes one or more of water, ethanol and methanol; the mass-volume ratio of the compound with the structure shown in Formula V to the third solvent is preferably 1g:(10-50)mL, specifically 1g:15mL, 1g:20mL, 1g:25mL, 1g:35mL or 1g:45mL.

[0103] In this invention, the temperature of the hydrolysis reaction is preferably 20-50°C, specifically 20°C, 30°C or 40°C, and the holding time is preferably 0.5-2h, specifically 1h or 1.5h.

[0104] In this invention, the hydrolysis reaction preferably includes sequentially adjusting the pH of the reaction solution to 5-6, allowing it to stand, and separating the solid and liquid components; the reagent used to adjust the pH of the reaction solution is preferably dilute hydrochloric acid with a mass concentration of 5%-20%; the standing time is preferably 1 hour, during which a white solid precipitates; and the solid-liquid separation is preferably performed by vacuum filtration.

[0105] After obtaining the compound with the structure shown in Formula VI, the present invention mixes the compound with the amino-substituted carboxylic acid ester, a condensing agent, and an organic solvent (denoted as the fourth organic solvent) to carry out a condensation reaction to obtain the compound with the structure shown in Formula VII. In the present invention, the amino-substituted carboxylic acid ester preferably includes one or more of methyl p-aminobenzoate, methyl p-aminomethylbenzoate, methyl 5-aminopyridine-2-carboxylate, methyl 7-aminoheptanoate, and methyl 5-aminopyridine-2-carboxylate; the molar ratio of the compound with the structure shown in Formula VI to the amino-substituted carboxylic acid ester is preferably 1:0.8 to 1.2, specifically 1:1.

[0106] In this invention, the condensing agent preferably includes one or more of HOBt, EDCI and HATU; the molar ratio of the compound with the structure shown in Formula VI to the condensing agent is preferably 1:1 to 3, specifically 1:1.5, 1:2 or 1:3.

[0107] In this invention, the fourth organic solvent preferably includes one or two of acetonitrile and tetrahydrofuran; the mass-volume ratio of the compound with the structure shown in Formula VI to the fourth organic solvent is preferably 1g:(5-50)mL, specifically 1g:10mL, 1g:15mL, 1g:25mL, 1g:30mL or 1g:40mL.

[0108] In this invention, the temperature of the condensation reaction is preferably room temperature, and the reaction time is preferably 6 to 10 hours, specifically 8 hours.

[0109] In this invention, the condensation reaction preferably further includes sequentially concentrating the reaction solution, redissolving the residue, washing the organic layer with water, washing with saturated brine, drying, solid-liquid separation, and purification; the reagents used for redissolving are preferably water and ethyl acetate; the volume ratio of water to ethyl acetate is preferably 3:2; the number of times the organic layer is washed with water is preferably 2 or more; the number of times the saturated brine is washed is preferably 1 or more; the drying is preferably drying with anhydrous sodium sulfate; the solid-liquid separation is preferably vacuum filtration; and the purification is preferably column chromatography purification.

[0110] After obtaining the compound with the structure shown in Formula VII, the present invention performs a hydroxylamine hydrolysis reaction by mixing the compound with hydroxylamine, water, a basic compound (denoted as the fourth basic compound), and an organic solvent (denoted as the fifth organic solvent) to obtain the compound based on the 6-methylfurano[2,3-d]pyrimidine-4(3H)-one structure. In the present invention, the hydroxylamine and water are preferably used as an aqueous solution of hydroxylamine; the mass ratio of hydroxylamine to water is preferably 1:1 to 10, specifically 1:1, 1:2, 1:4, 1:6, or 1:8.

[0111] In this invention, the molar ratio of the compound with the structure shown in Formula VII to hydroxylamine is preferably 1:20 to 100, specifically 1:40, 1:60 or 1:80.

[0112] In this invention, the fourth basic compound preferably includes one or more of sodium hydroxide and potassium hydroxide; the molar ratio of the compound with the structure shown in Formula VII to the fourth basic compound is preferably 1:5 to 20, specifically 1:6, 1:8, 1:10, 1:12, 1:15 or 1:18.

[0113] In this invention, the fifth organic solvent is preferably an alcohol; the alcohol includes one or more of methanol and ethanol; the mass-volume ratio of the compound with the structure shown in Formula VII to the fifth organic solvent is preferably 1g:(5-50)mL, specifically 1g:5mL, 1g:15mL, 1g:25mL, 1g:30mL, 1g:35mL or 1g:45mL.

[0114] In this invention, the temperature of the hydroxylamine hydrolysis reaction is preferably room temperature, and the reaction time is preferably 0.5 to 5 hours, specifically 0.5 hours, 1 hour, 3 hours or 4 hours.

[0115] In this invention, the reaction of hydroxylamine preferably further includes sequentially removing methanol from the reaction solution, adjusting the pH of the reaction solution to 7-8, allowing it to stand, and separating the solid and liquid components; the removal of methanol from the reaction solution is preferably carried out by vacuum distillation; the reagent used to adjust the pH of the reaction solution is preferably dilute hydrochloric acid with a mass concentration of 5%-10%; and the solid and liquid separation is preferably carried out by vacuum filtration.

[0116] The present invention also provides the use of compounds based on the 6-methylfurano[2,3-d]pyrimidine-4(3H)-one structure as described in the above-described scheme, or compounds based on the 6-methylfurano[2,3-d]pyrimidine-4(3H)-one structure obtained by the preparation method described in the above-described scheme, in the preparation of medicaments for treating diseases related to DNA methyltransferase 3A or histone deacetylase.

[0117] In this invention, the diseases associated with DNA methyltransferase 3A or histone deacetylase preferably include cancer.

[0118] To further illustrate the present invention, the following detailed description of the embodiments is provided in conjunction with the present invention, but these descriptions should not be construed as limiting the scope of protection of the present invention.

[0119] Example 1: Synthesis of N-[6-(hydroxycarbamoyl)pyridin-3-yl]-6-methyl-4-oxo-3-[3-(piperidin-1-yl)propyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (YQ-1A). The synthetic route is as follows:

[0120]

[0121] Ethyl 2-chloro-3-oxobutyrate (20 g, 121.32 mmol), malononitrile (8 g, 121.32 mmol), and 120 mL of ethanol were added to a three-necked flask at -10 °C. Then, 24 mL of 10% sodium ethoxide solution (133.45 mmol) was slowly added dropwise with stirring. After the addition was complete, the mixture was reacted at room temperature for 6 h. The reaction solution was concentrated, and 12 mL of petroleum ether and 4 mL of ethyl acetate were added to the residue. The mixture was stirred for 1 h and filtered to give ethyl 5-amino-4-cyano-2-methylfuran-3-carboxylic acid (II). 19.6 g of pale yellow solid was obtained, with a yield of 85.21%. mp: 78–79 °C. ESI-MS (m / z): 194.07 [M+Na] + .

[0122] At room temperature, ethyl 5-amino-4-cyano-2-methylfuran-3-carboxylate (II) (5.3 g, 27.29 mmol) and formic acid (18 mL, 500 mmol) were added to a three-necked flask. Acetic anhydride (15 mL, 168 mmol) was then slowly added dropwise to the reaction solution with stirring. After the addition was complete, the mixture was refluxed for 48 h. The reaction solution was concentrated, and the residue was mixed with water (30 mL). The pH was adjusted to 7 with a saturated sodium carbonate aqueous solution, resulting in a large amount of black solid. The mixture was stirred at room temperature for 2 h and filtered to obtain ethyl 6-methyl-4-oxo-1,4-dihydrofuran[2,3-d]pyrimidine-5-carboxylate (III), 4.6 g of crude black product, yield 75.41%; mp: 129–134 °C. 1H NMR (400MHz, DMSO-d6) δ12.58(s,1H),8.11(s,1H),4.27(q,J=7.1Hz,2H),2.58(s,3H),1.30(t,J=7.1Hz,3H).

[0123] At room temperature, ethyl 6-methyl-4-oxo-1,4-dihydrofuran[2,3-d]pyrimidine-5-carboxylate (III) (1.3 g, 5.90 mmol), 1,2-dibromoethane (6.62 g, 35.40 mmol), cesium carbonate (2.88 g, 8.85 mmol), and DMF (15 mL) were added to a three-necked flask and reacted at 25 °C for 8 h. The reaction solution was concentrated, ethyl acetate (20 mL) was added, the ethyl acetate layer was washed with water (20 mL × 3), washed once with saturated brine (25 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give 4.2 g of a viscous black solid. The solid was purified by column chromatography to give ethyl 3-(2-bromoethyl)-6-methyl-4-oxo-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxylate (IV), 1.1 g of white solid, yield 55%; mp: 45–46 °C.

[0124] At room temperature, ethyl 3-(2-bromoethyl)-6-methyl-4-oxo-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxylate (IV) (0.3 g, 0.91 mmol), piperidine (0.08 mL, 1.00 mmol), potassium carbonate (0.37 g, 2.73 mmol), and acetonitrile (10 mL) were added to a three-necked flask and reacted at 25 °C for 5 h. The reaction solution was concentrated, dissolved in dichloromethane (15 mL), washed twice with water (10 mL × 2), washed once with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give ethyl 6-methyl-4-oxo-3-[2-(piperidin-1-yl)ethyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxylate (V), 0.27 g white solid, yield 85.71%; mp: 97–100 °C.

[0125] At room temperature, ethyl 6-methyl-4-oxo-3-[2-(piperidin-1-yl)ethyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxylic acid (V) (0.2 g, 0.62 mmol) and 1M NaOH aqueous solution (5 mL) were added to a three-necked flask. The reaction was carried out at 50 °C for 1 h. 10% dilute hydrochloric acid was slowly added dropwise to the reaction solution until the pH was 5-6. After standing for 1 h, a white solid precipitated. The solid was filtered to give 0.14 g of 6-methyl-4-oxo-3-[2-(piperidin-1-yl)ethyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxylic acid (VI), yield 82.35%; mp: 119-123 °C.

[0126] At room temperature, 6-methyl-4-oxo-3-[2-(piperidin-1-yl)ethyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxylic acid (VI) (0.3 g, 0.94 mmol), methyl 5-aminopyridine-2-carboxylate (0.14 g, 0.94 mmol), EDCI (0.27 g, 1.41 mmol), HOBt (0.19 g, 1.41 mmol), and acetonitrile (10 mL) were added to a round-bottom flask. Then, TEA (0.29 mL, 2.07 mmol) was added dropwise at 0 °C. After the addition was complete, the mixture was transferred to room temperature and reacted for 8 h. The reaction solution was concentrated, and 15 mL of water and EA10 were added. mL, separate the EA layer, wash twice with water (15 mL × 2), wash once with saturated saline (15 mL), dry with anhydrous sodium sulfate, filter and concentrate the filtrate to obtain 0.38 g of oily crude product, purified by column chromatography to obtain methyl 5-{6-methyl-4-oxo-3-[3-(piperidin-1-yl)propyl]-3,4-dihydrofuran[2,3-d]pyrimidin-5-carboxamide}pyridine-2-carboxylate (VII), 0.15 g of white solid, yield 35.71%.

[0127] At room temperature, methyl 5-{6-methyl-4-oxo-3-[3-(piperidin-1-yl)propyl]-3,4-dihydrofuran[2,3-d]pyrimidin-5-carboxamide}pyridinecarboxylate (VII) (0.15 g, 0.33 mmol) and methanol (5 mL) were added to a single-necked flask. A 50% aqueous solution of hydroxylamine (2 mL) was slowly added dropwise at 0 °C, followed by a slow addition of 1 M NaOH aqueous solution (2 mL). After the addition was complete, the flask was transferred to room temperature and reacted for 0.5 h. The reaction mixture was then evaporated under reduced pressure. The methanol in the solution was added, and the pH of the reaction solution was adjusted to 7-8 with 10% dilute hydrochloric acid. After standing, the precipitated white solid was filtered to obtain N-[6-(hydroxycarbamoyl)pyridin-3-yl]-6-methyl-4-oxo-3-[3-(piperidin-1-yl)propyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (YQ-1A), 0.04 g, yield 28.57%; mp: 181-184℃; ESI-MS (m / z): 455.2 [M+H] + .

[0128] 1H NMR (400MHz, DMSO-d6) δ12.70(s,1H),11.32(s,1H),9.03(s,1H),8.92(d,J=2.4Hz,1H),8.62(s,1H),8.20(dd,J=8.6,2.5Hz,1H), 8.02(d,J=8.5Hz,1H),4.18(t,J=6.6Hz,2H),2.79(s,3H),2.29(dd,J=17.1,10.8Hz,6H),1.93(t,J=6.5Hz,2H),1.48-1.28(m,6H).

[0129] 13 C NMR(101MHz,DMSO-d6)δ162.95,161.58,160.70,160.33,159.92,150.42,145.51,139.44, 138.08,127.15,123.15,112.29,103.87,55.73,54.24,46.49,25.94,25.12,24.54,14.19.

[0130] Example 2: Synthesis of N-[6-(hydroxycarbamoyl)pyridin-3-yl]-6-methyl-4-oxo-3-[3-(pyrrolidin-1-yl)propyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (YQ-2A)

[0131] The preparation method in this embodiment is the same as in Example 1, except that piperidine is replaced with pyrrolidine to obtain the target compound YQ-2A; 0.06 g of white solid, yield 34.29%; mp: 190–192 °C; ESI-MS (m / z): 441.2 [M+H] + .

[0132] 1 H NMR (400MHz, DMSO-d6) δ12.68(s,1H),11.33(s,1H),9.04(s,1H),8.93(d,J=2.4Hz,1H),8.62(s,1H),8.22(dd,J=8.5,2.5Hz,1H) ,8.03(d,J=8.5Hz,1H),4.20(t,J=6.9Hz,2H),2.80(s,3H),2.48-2.36(m,6H),1.94(p,J=7.0Hz,2H),1.64(q,J=4.0,3.3Hz,4H).

[0133] 13C NMR (101MHz, DMSO-d6) δ162.95,161.59,160.71,160.29,159.95,150.35,145.53,139. 47,138.07,127.19,123.17,112.30,103.87,53.75,52.73,46.37,27.42,23.57,14.20.

[0134] Example 3: Synthesis of N-[6-(hydroxycarbamoyl)pyridin-3-yl]-6-methyl-4-oxo-3-[2-(pyrrolidone-1-yl)ethyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (FQX-2A)

[0135] The preparation method in this embodiment is the same as in Example 1, except that piperidine is replaced with pyrrolidine and 1,3-dibromopropane is replaced with 1,2-dibromoethane, yielding the target compound FQX-2A; 0.06 g of white solid, yield 34.29%; mp: 228–230 °C; ESI-MS (m / z): 425.0 [MH] - .

[0136] 1 H NMR (400MHz, DMSO-d6) δ12.61(s,1H),11.30(s,1H),8.92(s,1H),9.00-8.80(m,1H),8.56(s,1H),8.20(dd,J=8.8,2.3Hz,1H ), 8.01 (d, J = 8.6Hz, 1H), 4.23 (d, J = 6.9Hz, 2H), 2.80 (d, J = 4.7Hz, 5H), 2.54 (s, 2H), 1.86 (s, 2H), 1.67 (q, J = 4.3, 2.6Hz, 4H).

[0137] 13 C NMR(101MHz,DMSO-d6)δ161.80,160.43,159.55,158.94,158.88,149.23,144.44,1 38.34,136.91,126.06,122.01,111.19,102.78,52.98,52.95,44.99,22.61,13.11.

[0138] Example 43 Synthesis of 3-[3-(diethylamino)propyl]-N-[6-(hydroxycarbamoyl)pyridin-3-yl]-6-methyl-4-oxo-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (YQ-7A)

[0139] The preparation method in this embodiment is the same as in Example 1, except that piperidine is replaced with diethylamine to obtain the target compound YQ-7A; 0.05 g of white solid, yield 30.69%; mp: 215-217℃; ESI-MS (m / z): 443.2 [M+H] + .

[0140] 1 H NMR (400MHz, DMSO-d6) δ12.64(s,1H),11.32(s,1H),9.04(s,1H),8.92(d,J=2.4Hz,1H),8.66(s,1H),8.22(dd,J=8.6,2. 5Hz, 1H), 8.02 (d, J = 8.5Hz, 1H), 4.20 (t, J = 7.0Hz, 2H), 2.90-2.63 (m, 9H), 2.00 (d, J = 20.0Hz, 2H), 1.03 (d, J = 8.8Hz, 6H).

[0141] 13 C NMR (101MHz, DMSO-d6) δ162.92,161.58,160.68,160.26,160.01,150.21,145.54,139. 47,138.05,127.15,123.16,112.33,104.02,49.14,46.46,45.77,40.68,14.21,10.64.

[0142] Example 5: Synthesis of N-[4-(hydroxycarbamoyl)phenyl]-6-methyl-4-oxo-3-[3-(piperidin-1-yl)propyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (YQ-1B)

[0143] The preparation method in this embodiment is the same as in Example 1, yielding 6-methyl-4-oxo-3-[2-(piperidin-1-yl)ethyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxylic acid, which is then condensed with methyl 4-aminobenzoate to obtain methyl 4-{6-methyl-4-oxo-3-[3-(piperidin-1-yl)propyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamidobenzoate, followed by hydroxylamine hydrolysis to obtain the target compound YQ-1B. The specific steps are as follows:

[0144] At room temperature, 6-methyl-4-oxo-3-[2-(piperidin-1-yl)ethyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxylic acid (0.3 g, 0.94 mmol), methyl 4-aminobenzoate (0.14 g, 0.94 mmol), EDCI (0.27 g, 1.41 mmol), HOBt (0.19 g, 1.41 mmol), and ACN (10 mL) were added to a round-bottom flask. TEA (0.29 mL, 2.07 mmol) was added dropwise at 0 °C. After the addition was complete, the mixture was transferred to room temperature and reacted for 8 h. The reaction solution was concentrated, and 15 mL of water and 10 mL of EA were added. The EA layer was separated, washed twice with water (15 mL × 2), washed once with saturated brine (15 mL), dried over anhydrous sodium sulfate, and the filtrate was concentrated by filtration to obtain 0.32 g of oil. The oil was purified by PTLC to give methyl 4-{6-methyl-4-oxo-3-[3-(piperidin-1-yl)propyl]-3,4-dihydrofuran[2,3-d]pyrimidin-5-carboxamidobenzoate, 0.25 g of white solid, with a yield of 59.52%.

[0145] At room temperature, methyl 4-{6-methyl-4-oxo-3-[3-(piperidin-1-yl)propyl]-3,4-dihydrofuran[2,3-d]pyrimidin-5-carboxamide}benzoate (0.25 g, 0.55 mmol) and methanol (5 mL) were added to a round-bottom flask. A 50% hydroxylamine aqueous solution (2 mL) was slowly added dropwise at 0 °C, followed by a 1M NaOH aqueous solution (2 mL). After the addition was complete, the mixture was transferred to room temperature and reacted for 0.5 h. The methanol in the reaction solution was evaporated under reduced pressure, and the pH of the reaction solution was adjusted to 7–8 with 10% dilute hydrochloric acid. After standing for 3 h, a white solid precipitated. The solid was filtered to obtain 0.18 g of the target compound YQ-1B, with a yield of 72.29%; mp: 184–188 °C; ESI-MS (m / z): 454.1 [M+H] + .

[0146] 1 H NMR(400MHz,DMSO-d6)δ12.53(s,1H),11.15(s,1H),8.97(s,1H),8.60(s,1H),7.88-7.68(m,4H),4.18 (t,J=6.7Hz,2H),2.79(s,3H),2.29(dd,J=16.5,10.1Hz,6H),1.93(t,J=6.6Hz,2H),1.45-1.28(m,6H).

[0147] 13C NMR(101MHz,DMSO-d6)δ164.26,162.92,160.33,159.54,150.31,141.79,128.51,128 .22,118.95,112.67,103.90,82.77,55.72,54.25,46.43,25.94,25.13,24.55,14.19.

[0148] Example 6: Synthesis of N-[4-(hydroxycarbamoyl)phenyl]-6-methyl-4-oxo-3-[3-(pyrrolidone-1-yl)propyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (YQ-2B)

[0149] The preparation method in this embodiment is the same as in Example 5, except that piperidine is replaced with pyrrolidine to obtain the target compound YQ-2B; 0.12 g of white solid, yield 47.26%; mp: 187–190 °C; ESI-MS (m / z): 440.2 [M+H] + .

[0150] 1 H NMR(400MHz,DMSO-d6)δ12.63-12.27(m,1H),11.17(s,1H),9.00(s,1H),8.61(s,1H),7.84-7.69(m,4H), 4.20(t,J=6.8Hz,2H),2.98-2.81(m,6H),2.79(d,J=1.9Hz,3H),2.14-2.00(m,2H),1.77(d,J=6.0Hz,4H).

[0151] 13 C NMR (101MHz, DMSO-d6) δ164.17,162.91,160.28,159.55,150.24,141.60,129.19,128. 42,118.96,118.77,112.68,103.87,53.75,52.75,46.33,29.45,27.47,23.58,14.19.

[0152] Example 73 Synthesis of 3-[3-(dimethylamino)propyl]-N-[4-(hydroxycarbamoyl)phenyl]-6-methyl-4-oxo-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (YQ-9B)

[0153] The preparation method in this embodiment is the same as in Example 5, except that piperidine is replaced with dimethylamine to obtain the target compound YQ-9B; 0.09 g of white solid, yield 43.96%; mp: 159–162 °C; ESI-MS (m / z): 414.1 [M+H] + .

[0154] 1 H NMR(400MHz,DMSO-d6)δ12.41(s,1H),10.04(s,1H),8.57(s,1H),7.86-7.60(m,4H),4.14 (t,J=7.1Hz,2H),2.77(s,3H),2.26(t,J=6.7Hz,2H),2.13(s,6H),1.89(p,J=6.8Hz,2H).

[0155] 13 C NMR(101MHz,DMSO-d6)δ164.09,162.84,160.21,160.18,159.51,150.10,141.2 9,129.04,128.25,118.89,112.70,103.93,56.15,46.02,45.43,26.47,14.18.

[0156] Example 83: Synthesis of 3-[3-(diethylamino)propyl]-N-[4-(hydroxycarbamoyl)phenyl]-6-methyl-4-oxo-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (YQ-7B)

[0157] The preparation method in this embodiment is the same as in Example 5, except that piperidine is replaced with diethylamine to obtain the target compound YQ-7B; 0.12 g of white solid, yield 46.30%; mp: 151-152℃; ESI-MS (m / z): 442.1 [M+H] + .

[0158] 1 H NMR(400MHz,DMSO-d6)δ12.49(s,1H),11.16(s,1H),9.02(s,1H),8.59(s,1H),7.82-7.74(m,4H) ,4.15(t,J=7.0Hz,2H),2.78(s,3H),2.48-2.40(m,6H),1.92-1.87(m,2H),0.92(t,J=7.1Hz,6H).

[0159] Example 93: Synthesis of 4-(diethylamino)butyl]-N-[4-(hydroxycarbamoyl)phenyl]-6-methyl-4-oxo-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (WZ-7B)

[0160] The preparation method in this embodiment is the same as in Example 5, except that piperidine is replaced with diethylamine and 1,3-dibromopropane is replaced with 1,4-dibromobutane, yielding the target compound WZ-7B; 0.06 g of white solid, yield 34.26%; mp: 134–136 °C; ESI-MS (m / z): 456.2 [M+H] + .

[0161] 1 H NMR (400MHz, DMSO-d6) δ12.46(s,1H),11.11(s,1H),9.04(s,1H),8.63(s,1H),7.78(q,J=8.7Hz,4H),4.13(t,J=7.2H z, 2H), 2.78 (s, 3H), 2.41 (dq, J = 13.5, 7.1Hz, 6H), 1.75 (p, J = 7.2Hz, 2H), 1.43 (p, J = 7.2Hz, 2H), 0.93 (t, J = 7.1Hz, 6H).

[0162] 13 C NMR (101MHz, DMSO-d6) δ164.24,162.82,160.27,160.13,159.66,149.87,141.75,128. 48,128.22,118.95,112.67,104.01,52.24,47.40,46.75,27.30,24.25,14.20,12.23.

[0163] Example 10 Synthesis of N-[4-(hydroxycarbamoyl)phenyl]-6-methyl-4-oxo-3-[4-(piperidin-1-yl)butyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (WZ-1B)

[0164] The preparation method in this embodiment is the same as in Example 5, except that 1,3-dibromopropane is replaced with 1,4-dibromobutane to obtain the target compound WZ-1B; 0.07 g of white solid, yield 39.33%; mp: 177–179 °C; ESI-MS (m / z): 468.2 [M+H] + .

[0165] 1H NMR(400MHz,DMSO-d6)δ12.40(s,1H),11.19(s,1H),8.99(s,1H),8.68(s,1H),7.88-7.50(m,4H), 4.14(d,J=8.1Hz,2H),3.20-2.87(m,6H),2.77(s,3H),1.75(dd,J=12.1,6.2Hz,8H),1.51(s,2H).

[0166] 13 C NMR(101MHz,DMSO-d6)δ164.23,162.81,160.22,159.67,149.94,141.71,131.14,128.51, 128.22,118.93,112.67,104.03,55.83,52.54,46.77,26.38,23.03,22.12,20.86,14.19.

[0167] Example 11 Synthesis of N-[4-(hydroxycarbamoyl)phenyl]-6-methyl-4-oxo-3-[2-(pyrrolidone-1-yl)ethyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (FQX-1B)

[0168] The preparation method in this embodiment is the same as in Example 1, yielding 6-methyl-4-oxo-3-[2-(pyrrolidone-1-yl)ethyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxylic acid, which is then condensed with 4-amino-N-(benzyloxy)benzamide to obtain N-{4-[(benzyloxy)carbamoyl]phenyl}-6-methyl-4-oxo-3-(2-(pyrrolidone-1-yl)ethyl)-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide, followed by hydroxylamine hydrolysis to obtain the target compound FQX-1B. The specific steps are as follows:

[0169] At room temperature, 0.26 g (0.91 mmol) of 6-methyl-4-oxo-3-[2-(pyrrolidone-1-yl)ethyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxylic acid, 4-amino-N-(benzyloxy)benzamide, 0.25 g (0.91 mmol), HATU (0.52 g (1.37 mmol), and acetonitrile (10 mL) were added to a three-necked flask. Then, DIPEA (0.47 mL (2.73 mmol) was slowly added dropwise at 0 °C. After the addition was complete, the mixture was transferred to a heated magnetic stirrer and reacted at 50 °C. 10 h; concentrate the reaction solution, add 15 mL of ethyl acetate, wash the ethyl acetate layer with water (15 mL × 2), wash once with saturated brine (15 mL), dry with anhydrous sodium sulfate, filter, concentrate under reduced pressure to obtain 0.25 g of crude product, purified by column chromatography to obtain N-{4-[(benzyloxy)carbamoyl]phenyl}-6-methyl-4-oxo-3-(2-(pyrrolidine-1-yl)ethyl)-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide; white solid 0.12 g, yield 38.71%; mp: 189~193℃.

[0170] At room temperature, N-{4-[(benzyloxy)carbamoyl]phenyl}-6-methyl-4-oxo-3-(2-(pyrrolidone-1-yl)ethyl)-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (0.1 g, 0.29 mmol), palladium on carbon (0.01 g, 0.029 mmol), and 6 mL of a mixture of methanol and ethyl acetate (v / v, 1:1) were added to a 50 mL round-bottom flask, and the mixture was purged with hydrogen for 4 h. The palladium on carbon was removed by filtration, and the filtrate was concentrated under reduced pressure to give the target compound FQX-1B: 0.05 g white solid, yield 62.50%; mp: 209–211 °C; ESI-MS (m / z): 426.2 [M+H] + .

[0171] 1 H NMR(400MHz,DMSO-d6)δ12.43(s,1H),11.16(s,1H),8.97(s,1H),8.54(s,1H),7.84-7.7 3(m,4H),4.24(t,J=6.1Hz,2H),2.84-2.79(m,5H),2.55(d,J=17.7Hz,4H),1.69(s,4H).

[0172] 13C NMR(101MHz,DMSO-d6)δ163.21,161.77,159.20,158.99,158.58,149.13,140.6 6,127.44,127.16,117.89,111.58,102.83,52.96,39.45,30.62,22.59,13.13.

[0173] Example 1: Synthesis of 2N-[4-(hydroxycarbamoyl)phenyl]-6-methyl-4-oxo-3-[2-(piperidin-1-yl)ethyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (FQX-2B)

[0174] The preparation method in this embodiment is the same as in Example 11, except that pyrrolidine is replaced with piperidine to obtain the target compound FQX-2B; 0.04 g of white solid, yield 44.09%; mp: 183-185℃; ESI-MS (m / z): 440.1 [M+H] + .

[0175] 1 H NMR(400MHz,DMSO-d6)δ12.44(s,1H),11.16(s,1H),8.96(s,1H),8.49(s,1H),7 .86-7.71(m,4H),4.23(s,2H),2.78(s,3H),2.43(s,6H),1.42(d,J=32.9Hz,6H).

[0176] 13 C NMR(101MHz,DMSO-d6)δ164.31,162.85,160.29,160.07,159.59,150.34,141.76,1 28.52,128.22,118.96,118.76,112.67,103.77,56.64,54.36,26.02,24.24,14.20.

[0177] Example 13 Synthesis of N-[4-(hydroxycarbamoyl)benzyl]-6-methyl-4-oxo-3-[2-(pyrrolidone-1-yl)ethyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (FQX-1C)

[0178] The preparation method in this embodiment is the same as in Example 1, yielding 6-methyl-4-oxo-3-[2-(pyrrolidone-1-yl)ethyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxylic acid, which is then condensed with methyl 4-(aminomethyl)benzoate to obtain methyl 4-({6-methyl-4-oxo-3-[2-(pyrrolidone-1-yl)ethyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide}methyl)benzoate, followed by hydroxylamine hydrolysis to obtain the target compound FQX-1C. The specific steps are as follows:

[0179] At room temperature, 6-methyl-4-oxo-3-[2-(pyrrolidone-1-yl)ethyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxylic acid (0.3 g, 0.94 mmol), methyl 4-(aminomethyl)benzoate (0.16 g, 0.94 mmol), EDCI (0.27 g, 1.41 mmol), HOBt (0.19 g, 1.41 mmol), and ACN (10 mL) were added to a round-bottom flask. TEA (0.29 mL, 2.07 mmol) was added dropwise at 0 °C. After the addition was complete, the mixture was transferred to room temperature and reacted for 8 h. The reaction solution was concentrated, and 15 mL of water was added. 10 mL was used to separate the EA layer, which was washed twice with water (15 mL × 2), once with 15 mL of saturated saline solution (15 mL), dried over anhydrous sodium sulfate, and the filtrate was concentrated by filtration to obtain 0.43 g of oily crude product. The product was purified by column chromatography to obtain methyl 4-({6-methyl-4-oxo-3-[2-(pyrrolidone-1-yl)ethyl]-3,4-dihydrofuran[2,3-d]pyrimidin-5-carboxamide}methyl)benzoate, 0.35 g of white solid, with a yield of 85.37%.

[0180] At room temperature, methyl 4-({6-methyl-4-oxo-3-[2-(pyrrolidone-1-yl)ethyl]-3,4-dihydrofuran[2,3-d]pyrimidin-5-carboxamide}methyl)benzoate (0.35 g, 0.80 mmol) and methanol (5 mL) were added to a round-bottom flask. A 50% hydroxylamine aqueous solution (2 mL) was slowly added dropwise at 0 °C, followed by a 1M NaOH aqueous solution (2 mL). After the addition was complete, the mixture was transferred to room temperature and allowed to react for 0.5 h. The methanol in the reaction solution was evaporated under reduced pressure at 30 °C. The pH of the reaction solution was adjusted to 7–8 with 10% dilute hydrochloric acid. After standing for 3 h, a large amount of white solid precipitated. The solid was filtered to obtain the target compound FQX-1C, 0.30 g, with a yield of 85.71%; mp: 206–209 °C; ESI-MS (m / z): 440.2 [M+H]. + .

[0181] 1H NMR(400MHz,DMSO-d6)δ11.16(s,1H),10.54(t,J=5.8Hz,1H),9.00(s,1H),8.47(s,1H),7.75-7.68(m,2H),7 .43-7.37(m,2H),4.58(d,J=5.8Hz,2H),4.14(t,J=6.1Hz,2H),2.72(d,J=11.3Hz,5H),1.65(p,J=3.1Hz,4H).

[0182] Example 14 Synthesis of N-[4-(hydroxycarbamoyl)benzyl]-6-methyl-4-oxo-3-[2-(piperidin-1-yl)ethyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (FQX-2C)

[0183] The preparation method in this embodiment is the same as in Example 13, except that pyrrolidine is replaced with piperidine to obtain the target compound FQX-2C; 0.26 g of white solid, yield 89.35%; mp: 180–182 °C; ESI-MS (m / z): 454.2 [M+H] + .

[0184] 1 H NMR(400MHz,DMSO-d6)δ11.17(s,1H),10.54(s,1H),8.99(s,1H),8.43(s,1H),7.74-7.68(m,2H),7.43-7.3 7(m,2H),4.57(d,J=5.7Hz,2H),4.14(s,2H),2.74(s,3H),2.57(s,2H),2.38(s,4H),1.39(d,J=26.3Hz,6H).

[0185] Example 15 Synthesis of N-[4-(hydroxycarbamoyl)benzyl]-3-[2-(isopropylamino)ethyl]-6-methyl-4-oxo-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (FQX-3C)

[0186] The preparation method in this embodiment is the same as in Example 13, except that pyrrolidine is replaced with isopropylamine to obtain the target compound FQX-3C; 0.16 g of white solid, yield 65.23%; mp: 222-224℃; ESI-MS (m / z): 426.0 [MH] - .

[0187] 1H NMR (400MHz, DMSO-d6) δ10.58(s,1H),8.45(s,1H),7.75-7.66(m,2H),7.38(d,J=8.1Hz,2H),4.56(d,J=5.8 Hz, 2H), 4.05 (t, J = 6.0Hz, 2H), 2.78 (t, J = 6.0Hz, 2H), 2.73 (s, 3H), 2.68-2.64 (m, 1H), 0.90 (d, J = 6.2Hz, 6H).

[0188] Example 1: Synthesis of 6N-[4-(hydroxycarbamoyl)benzyl]-6-methyl-3-[2-(4-methylpiperazin-1-yl)ethyl]-4-oxo-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (FQX-5C)

[0189] The preparation method in this embodiment is the same as in Example 13, except that pyrrolidine is replaced with N-methylpiperazine to obtain the target compound FQX-5C; 0.27 g of white solid, yield 80.97%; mp: 155-157℃; ESI-MS (m / z): 469.2 [M+H] + .

[0190] 1 H NMR (400MHz, DMSO-d6) δ11.17(s,1H),10.55(t,J=5.8Hz,1H),9.01(s,1H),8.43(s,1H),7.72(d,J=8.1Hz,2H),7.39(d,J=8.1 Hz, 2H), 4.57 (d, J = 5.7Hz, 2H), 4.13 (t, J = 5.9Hz, 2H), 2.73 (s, 3H), 2.56 (t, J = 6.0Hz, 2H), 2.33 (d, J = 73.3Hz, 8H), 2.12 (s, 3H).

[0191] 13 C NMR(101MHz,DMSO-d6)δ162.25,161.08,158.85,157.56,149.06,141.73,130.87, 126.57,126.45,111.97,102.93,54.99,54.13,51.88,45.07,42.91,41.29,12.92.

[0192] Example 173 Synthesis of 3-[2-(diethylamino)ethyl]-N-[4-(hydroxycarbamoyl)benzyl]-6-methyl-4-oxo-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (FQX-7C)

[0193] The preparation method in this embodiment is the same as in Example 13, except that pyrrolidine is replaced with diethylamine to obtain the target compound FQX-7C; 0.19 g of white solid, yield 72.69%; mp: 205-207℃; ESI-MS (m / z): 442.1 [M+H] + .

[0194] 1 H NMR (400MHz, DMSO-d6) δ11.15(s,1H),10.58(t,J=5.8Hz,1H),9.01(s,1H),8.41(s,1H),7.71(d,J=8.0Hz,2H),7.39(d,J=8.0Hz,2 H), 4.57 (d, J = 5.7Hz, 2H), 4.07 (t, J = 5.8Hz, 2H), 2.73 (s, 3H), 2.63 (t, J = 5.8Hz, 2H), 2.44 (q, J = 7.1Hz, 4H), 0.80 (t, J = 7.0Hz, 6H).

[0195] 13 C NMR(101MHz,DMSO-d6)δ164.49,162.99,161.80,159.62,158.23,150.26,142.78,1 31.99,127.58,127.52,112.08,103.89,50.76,47.08,45.57,42.38,14.00,12.46.

[0196] Example 18 Synthesis of N-[4-(hydroxycarbamoyl)benzyl]-6-methyl-3-[3-(4-methylpiperazin-1-yl)propyl]-4-oxo-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (YQ-5C)

[0197] The preparation method in this embodiment is the same as in Example 13, except that pyrrolidine is replaced with N-methylpiperazine and 1,2-dibromoethane is replaced with 1,3-dibromopropane, yielding the target compound YQ-5C; 0.19 g of white solid, yield 69.63%; mp: 189–190 °C; ESI-MS (m / z): 483.3 [M+H] + .

[0198] 1H NMR (400MHz, DMSO-d6) δ11.16(s,1H),10.60(t,J=5.8Hz,1H),9.02(s,1H),8.53(s,1H),7.75-7.67(m,2H),7.39(d, J=8.1Hz,2H),4.57(d,J=5.8Hz,2H),4.08(t,J=6.7Hz,2H),2.73(s,3H),2.49-1.96(m,13H),1.85(p,J=6.6Hz,2H).

[0199] 13 C NMR(101MHz,DMSO-d6)δ164.44,162.92,161.82,159.90,158.11,149.96,142.83,131.95, 127.58,127.53,112.18,104.23,54.99,54.84,52.51,46.23,45.87,42.35,25.00,13.98.

[0200] Example 193: Synthesis of 3-[3-(diethylamino)propyl]-N-[4-(hydroxycarbamoyl)benzyl]-6-methyl-4-oxo-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (YQ-7C)

[0201] The preparation method in this embodiment is the same as in Example 13, except that pyrrolidine is replaced with diethylamine and 1,2-dibromoethane is replaced with 1,3-dibromopropane, yielding the target compound YQ-7C; 0.14 g of white solid, yield 56.77%; mp: 129–132 °C; ESI-MS (m / z): 456.3 [M+H] + .

[0202] 1 H NMR (400MHz, DMSO-d6) δ10.93(s,1H),10.58(t,J=5.8Hz,1H),9.08(s,1H),8.53(s,1H),7.78-7.63(m,2H),7.40(d,J=8.3Hz,2 H), 4.57 (d, J = 5.7Hz, 2H), 4.06 (t, J = 7.1Hz, 2H), 2.73 (s, 3H), 2.47-2.33 (m, 6H), 1.82 (p, J = 6.9Hz, 2H), 0.89 (t, J = 7.1Hz, 6H).

[0203] 13C NMR (101MHz, DMSO-d6) δ164.42,162.89,161.80,159.71,158.22,149.98,142.77,132. 01,127.59,127.51,112.12,104.15,49.68,46.31,46.17,42.37,26.11,13.99,11.80.

[0204] Example 20 Synthesis of N-[4-(hydroxycarbamoyl)benzyl]-3-(2-hydroxyethyl)-6-methyl-4-oxo-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (YQ-8C)

[0205] The preparation method in this embodiment is the same as in Example 13, except that 6-methyl-4-oxo-3-[2-(pyrrolidone-1-yl)ethyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxylic acid is replaced with 3-(2-hydroxyethyl)-6-methyl-4-oxo-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxylic acid, yielding the target compound YQ-8C; 0.06 g white solid, yield 25.63%; mp: 208–212 °C; ESI-MS (m / z): 387.1 [M+H] + .

[0206] 1 H NMR (400MHz, DMSO-d6) δ11.18(s,1H),10.56(t,J=5.8Hz,1H),9.00(s,1H),8.42(s,1H),7.72(d,J=8.0Hz,2H),7.40(d ,J=8.0Hz,2H),4.96(t,J=5.6Hz,1H),4.57(d,J=5.8Hz,2H),4.11(t,J=5.2Hz,2H),3.65(q,J=5.4Hz,2H),2.74(s,3H).

[0207] 13 C NMR(101MHz,DMSO-d6)δ164.57,163.01,161.78,159.73,158.22,150.30,14 2.81,132.01,127.65,127.57,112.11,104.16,58.40,49.76,42.39,14.00.

[0208] Example 2. Synthesis of 13-(2-(dimethylamino)ethyl)-N-(4-(hydroxycarbamoyl)benzyl)-6-methyl-4-oxo-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (FQX-9C)

[0209] The preparation method in this embodiment is the same as in Example 13, except that pyrrolidine is replaced with dimethylamine to obtain the target compound FQX-9C; 0.14 g of white solid, yield 51.63%; mp: 210-212℃; ESI-MS (m / z): 414.1 [M+H] + .

[0210] 1 H NMR (400MHz, DMSO-d6) δ11.17(s,1H),10.54(t,J=5.9Hz,1H),9.00(s,1H),8.45(s,1H),7.71(d,J=8.0Hz,2H),7. 40(d,J=8.0Hz,2H), 4.57(d,J=5.7Hz,2H), 4.12(t,J=6.0Hz,2H), 2.73(s,3H), 2.53(d,J=5.7Hz,2H), 2.17(s,6H).

[0211] 13 C NMR(101MHz,DMSO-d6)δ164.53,162.85,161.80,159.57,158.28,150.04,142.8 4,131.97,127.63,127.54,112.11,104.10,57.54,45.69,44.62,42.36,14.01.

[0212] Example 22 Synthesis of N-[4-(hydroxycarbamoyl)benzyl]-6-methyl-4-oxo-3-[3-(pyrrolidone-1-yl)propyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (YQ-1C)

[0213] The preparation method in this embodiment is the same as in Example 13, except that 1,2-dibromoethane is replaced with 1,3-dibromopropane to obtain the target compound YQ-1C; 0.21 g of white solid, yield 79.88%; mp: 128–131 °C; ESI-MS (m / z): 454.1 [M+H] + .

[0214] 1H NMR (400MHz, DMSO-d6) δ11.11(s,1H),10.59(t,J=5.8Hz,1H),9.06(s,1H),8.53(s,1H),7.72(d,J=7.9Hz,2H),7.39(d,J=8.0Hz,2H),4. 57(d,J=5.8Hz,2H),4.09(t,J=7.0Hz,2H),2.73(s,3H),2.38(dt,J=22.8,6.3Hz,6H),1.85(p,J=6.9Hz,2H),1.61(q,J=3.8,3.3Hz,4H).

[0215] 13 C NMR (101MHz, DMSO-d6) δ164.41,162.91,161.81,159.76,158.20,150.00,142.75,132. 02,127.60,127.49,112.10,104.10,53.73,52.76,46.17,42.36,27.52,23.55,14.00.

[0216] Example 23 Synthesis of N-[4-(hydroxycarbamoyl)benzyl]-6-methyl-4-oxo-3-[3-(piperidin-1-yl)propyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (YQ-2C)

[0217] The preparation method in this embodiment is the same as in Example 13, except that pyrrolidine is replaced with piperidine and 1,2-dibromoethane is replaced with 1,3-dibromopropane, yielding the target compound YQ-2C; 0.20 g of white solid, yield 74.98%; mp: 130–133 °C; ESI-MS (m / z): 468.2 [M+H] + .

[0218] 1 H NMR (400MHz, DMSO-d6) δ11.14(s,1H),10.61(t,J=5.8Hz,1H),9.02(s,1H),8.53(s,1H),7.71(d,J=8.0Hz,2H),7.39(d,J=8.0Hz,2H),4.5 7(d,J=5.8Hz,2H),4.07(t,J=6.7Hz,2H),2.73(s,3H),2.24(dd,J=16.1,9.7Hz,6H),1.84(p,J=6.7Hz,2H),1.33(tt,J=13.5,5.7Hz,6H).

[0219] 13C NMR(101MHz,DMSO-d6)δ164.43,162.94,161.83,159.85,158.15,150.07,142.80,132.00, 127.56,127.50,112.13,104.14,55.82,54.24,46.34,42.35,25.95,25.11,24.55,14.00.

[0220] Example 24 Synthesis of N-[4-(hydroxycarbamoyl)benzyl]-6-methyl-3-(3-morpholinopropyl)-4-oxo-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (YQ-4C)

[0221] The preparation method in this embodiment is the same as in Example 13, except that pyrrolidine is replaced with morpholine and 1,2-dibromoethane is replaced with 1,3-dibromopropane, yielding the target compound YQ-4C; 0.16 g of white solid, yield 51.79%; mp: 127–129 °C; ESI-MS (m / z): 470.2 [M+H] + .

[0222] 1 H NMR (400MHz, DMSO-d6) δ11.16(s,1H),10.59(t,J=5.8Hz,1H),9.02(d,J=22.8Hz,1H),8.55(s,1H),7.80-7.64(m,2H),7.39(d,J=8.1Hz,2 H), 4.57 (d, J = 5.8Hz, 2H), 4.09 (t, J = 6.8Hz, 2H), 3.43 (t, J = 4.6Hz, 4H), 2.73 (s, 3H), 2.28 (dt, J = 19.6, 5.6Hz, 6H), 1.86 (p, J = 6.7Hz, 2H).

[0223] 13 C NMR (101MHz, DMSO-d6) δ164.49,162.94,161.80,159.86,158.21,150.04,142.83,131. 96,127.61,127.52,112.13,104.17,66.55,55.48,53.49,46.19,42.36,24.79,14.00.

[0224] Example 253 Synthesis of 3-(4-ethylpiperazin-1-yl)propyl]-N-[4-(hydroxycarbamoyl)benzyl]-6-methyl-4-oxo-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (YQ-6C)

[0225] The preparation method in this embodiment is the same as in Example 13, except that pyrrolidine is replaced with N-ethylpiperazine and 1,2-dibromoethane is replaced with 1,3-dibromopropane, yielding the target compound YQ-6C; 0.17 g of white solid, yield 54.38%; mp: 108–110 °C; ESI-MS (m / z): 497.3 [M+H] + .

[0226] 1 H NMR (400MHz, DMSO-d6) δ11.16(s,1H),10.61(t,J=5.8Hz,1H),9.00(s,1H),8.52(s,1H),7.85-7.66(m,2H),7.49-7.31(m,2H) ,4.57(d,J=5.8Hz,2H),4.08(t,J=6.7Hz,2H),2.73(s,3H),2.43-1.98(m,12H),1.85(p,J=6.6Hz,2H),0.92(t,J=7.2Hz,3H).

[0227] 13 C NMR(101MHz,DMSO-d6)δ164.45,162.94,161.83,159.94,158.10,149.97,142.86,131.94,127 .57,127.51,112.19,104.24,55.20,52.95,52.61,52.03,46.33,42.34,24.96,13.97,12.23.

[0228] Example 26 Synthesis of N-[7-(hydroxyamino)-7-oxohepyl]-6-methyl-4-oxo-3-[3-(piperidin-1-yl)propyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (YQ-1D)

[0229] The preparation method in this embodiment is the same as in Example 1, yielding 6-methyl-4-oxo-3-[3-(piperidin-1-yl)propyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxylic acid. This is then condensed with methyl 7-aminoheptanoate hydrochloride to obtain methyl 7-{6-methyl-4-oxo-3-[3-(piperidin-1-yl)propyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide}heptanoate. Finally, hydroxylamine hydrolysis is performed to obtain the target compound YQ-1D. The specific steps are as follows:

[0230] At room temperature, 6-methyl-4-oxo-3-[3-(piperidin-1-yl)propyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxylic acid (0.3 g, 0.86 mmol), methyl 7-aminoheptanoate hydrochloride (0.17 g, 0.86 mmol), EDCI (0.25 g, 1.29 mmol), HOBt (0.17 g, 1.29 mmol), and ACN (10 mL) were added to a round-bottom flask. TEA (0.39 mL, 2.75 mmol) was added dropwise at 0 °C. After the addition was complete, the mixture was transferred to room temperature and reacted for 8 h. The reaction solution was concentrated, and 15 mL of water was added. 10 mL was used to separate the EA layer, which was washed twice with water (15 mL × 2), once with saturated saline (15 mL), dried over anhydrous sodium sulfate, and the filtrate was concentrated by filtration to obtain 0.56 g of oily crude product. The product was purified by column chromatography to obtain methyl 7-{6-methyl-4-oxo-3-[3-(piperidin-1-yl)propyl]-3,4-dihydrofuran[2,3-d]pyrimidin-5-carboxamide}heptanoate, a pale yellow solid, 0.36 g, with a yield of 91.14%.

[0231] At room temperature, methyl 7-{6-methyl-4-oxo-3-[3-(piperidin-1-yl)propyl]-3,4-dihydrofuran[2,3-d]pyrimidin-5-carboxamide}heptanoate (0.36 g, 0.78 mmol) and methanol (5 mL) were added to a round-bottom flask. A 50% hydroxylamine aqueous solution (3 mL) was slowly added dropwise at 0 °C, followed by a 1M NaOH aqueous solution (2 mL). After the addition was complete, the mixture was transferred to room temperature and allowed to react for 0.5 h. The methanol in the reaction solution was evaporated under reduced pressure, and the pH of the reaction solution was adjusted to 7–8 with 10% dilute hydrochloric acid. After standing for 3 h, a white solid precipitated. The solid was filtered to obtain a filter cake of 0.32 g, with a yield of 88.89%. mp: 94–96 °C; ESI-MS (m / z): 462.2 [M+H] + .

[0232] 1 H NMR (400MHz, DMSO-d6) δ10.30(s,1H),10.10(t,J=5.4Hz,1H),8.71-8.60(m,1H),8.51(s,1H),4.09(t,J=6.7Hz,2H),3.26(dd,J=7.1,5.5H z, 2H), 2.71 (s, 3H), 2.25 (dd, J = 15.5, 9.0Hz, 6H), 1.90 (dt, J = 30.1, 7.0Hz, 4H), 1.50 (h, J = 7.6Hz, 4H), 1.31 (dq, J = 29.6, 6.6, 6.2Hz, 10H).

[0233] 13C NMR(101MHz,DMSO-d6)δ169.55,162.88,161.50,159.80,157.63,149.98,112.44,104.14,5 5.80,54.23,46.31,38.96,32.69,29.32,28.72,26.63,25.94,25.52,25.12,24.55,13.93.

[0234] Example 27 Synthesis of N-[7-(hydroxyamino)-7-oxohepyl]-6-methyl-4-oxo-3-[2-(piperidin-1-yl)ethyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (FQX-1D)

[0235] The preparation method in this embodiment is the same as in Example 26, except that 1,3-dibromopropane is replaced with 1,2-dibromoethane to obtain the target compound FQX-1D; 0.30 g of white solid, yield 79.58%; mp: 88-89℃; ESI-MS (m / z): 448.2 [M+H] + .

[0236] 1 H NMR (400MHz, DMSO-d6) δ10.31(s,1H),10.04(t,J=5.4Hz,1H),8.63(s,1H),8.41(s,1H),4.14(t,J=6.0Hz,2H),3.29 -3.24(m,2H),2.71(s,3H),2.54(t,J=6.0Hz,2H),2.37(d,J=5.9Hz,4H),1.94(t,J=7.4Hz,2H),1.52-1.23(m,14H).

[0237] 13 C NMR(101MHz,DMSO-d6)δ169.56,162.83,161.47,159.52,157.71,150.05,112.43,103.98,56.80,54.44,44.21,39.00,32.68,29.33, 28.72,26.64,26.11,25.52,24.34,13.93.),2.54(t,J=6.0Hz,2H),2.37(d,J=5.9Hz,4H),1.94(t,J=7.4Hz,2H),1.52-1.23(m,14H).

[0238] Example 283 Synthesis of 3-[3-(diethylamino)propyl]-N-[7-(hydroxyamino)-7-oxohepyl]-6-methyl-4-oxo-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (YQ-7D)

[0239] The preparation method in this embodiment is the same as in Example 26, except that piperidine is replaced with ethylenediamine to obtain the target compound YQ-7D; 0.20 g of white solid, yield 65.81%; mp: 115–117 °C; ESI-MS (m / z): 450.3 [M+H] + .

[0240] 1 H NMR (400MHz, DMSO-d6) δ10.31(s,1H),10.08(t,J=5.3Hz,1H),8.66(s,1H),8.51(s,1H),4.07(t,J=7.0Hz,2H),3.27(td,J=6.9,5.3Hz,2H),2 .71(s,3H),2.47-2.34(m,6H),1.94(t,J=7.4Hz,2H),1.83(p,J=6.9Hz,2H),1.50(h,J=7.5Hz,4H),1.38-1.23(m,4H),0.90(t,J=7.1Hz,6H).

[0241] 13 C NMR(101MHz,DMSO-d6)δ169.54,162.84,161.47,159.67,157.70,149.89,112.43,104.1 5,49.69,46.32,46.16,38.98,32.69,29.32,28.73,26.64,26.09,25.51,13.92,11.82.

[0242] Example 293 Synthesis of 3-[3-(diethylamino)propyl]-N-[7-(hydroxyamino)-7-oxohepyl]-6-methyl-4-oxo-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (YQ-9D)

[0243] The preparation method in this embodiment is the same as in Example 26, except that piperidine is replaced with ethylmethylamine to obtain the target compound YQ-9D; 0.21 g of white solid, yield 69.34%; mp: 155-156℃; ESI-MS (m / z): 422.2 [M+H] + .

[0244] 1H NMR (400MHz, DMSO-d6) δ10.32(s,1H),10.06(t,J=5.3Hz,1H),8.67(s,1H),8.51(s,1H),4.07(t,J=7.1Hz,2H),3.26(s,2H),2.71(s,3H), 2.24(t,J=6.7Hz,2H),2.12(s,6H),1.94(t,J=7.4Hz,2H),1.84(p,J=6.9Hz,2H),1.50(h,J=7.6Hz,4H),1.30(dq,J=24.8,8.9,8.1Hz,4H).

[0245] 13 C NMR(101MHz,DMSO-d6)δ169.51,162.84,161.47,159.67,157.73,149.82,112.44,10 4.18,56.16,45.86,45.38,38.98,32.67,29.33,28.72,26.64,26.49,25.52,13.93.

[0246] Example 303 Synthesis of 3-[3-(diethylamino)propyl]-N-[7-(hydroxyamino)-7-oxohepyl]-6-methyl-4-oxo-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (WZ-1D)

[0247] The preparation method in this embodiment is the same as in Example 26, except that 1,3-dibromopropane is replaced with 1,4-dibromobutane to obtain the target compound WZ-1D; 0.15 g of white solid, yield 56.39%; mp: 108-109 °C; ESI-MS (m / z): 476.2 [M+H] + .

[0248] 1 H NMR (400MHz, DMSO-d6) δ10.31(s,1H),10.06(t,J=5.4Hz,1H),8.71(s,1H),8.55(s,1H),4.05(t,J=7.2Hz,2H),3.29-3. 21(m,2H),2.71(s,3H),2.25(dt,J=18.6,6.2Hz,6H),1.94(t,J=7.4Hz,2H),1.69(p,J=7.3Hz,2H),1.54-1.24(m,16H).

[0249] 13C NMR(101MHz,DMSO-d6)δ169.54,162.81,161.45,159.60,157.77,149.60,112.44,104.24,58.4 0,54.46,47.23,38.97,32.69,29.34,28.72,27.22,26.62,26.02,25.52,24.60,23.76,13.93.

[0250] Example 31 Synthesis of N-[7-(hydroxyamino)-7-oxohepyl]-6-methyl-4-oxo-3-[2-(pyrrolidone-1-yl)ethyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxamide (FQX-2D)

[0251] The preparation method in this embodiment is the same as in Example 26, except that piperidine is replaced with pyrrolidine and 1,3-dibromopropane is replaced with 1,2-dibromoethane to obtain the target compound FQX-2D. Using 6-methyl-4-oxo-3-[2-(pyrrolidine-1-yl)ethyl]-3,4-dihydrofuran[2,3-d]pyrimidine-5-carboxylic acid as the starting material, the target compound FQX-2D is obtained through a two-step reaction. 0.33 g of white solid was obtained, with a yield of 94.10%. mp: 183–185 °C; ESI-MS (m / z): 434.2 [M+H]. + .

[0252] 1 H NMR (400MHz, DMSO-d6) δ10.32(s,1H),10.03(t,J=5.4Hz,1H),8.65(s,1H),8.46(s,1H),4.15(t,J=6.1Hz,2H),3.27(q,J=6.8Hz ,2H),2.72(d,J=7.1Hz,5H),2.51(s,4H),1.94(t,J=7.4Hz,2H),1.67(h,J=3.1Hz,4H),1.49(p,J=7.4Hz,4H),1.38-1.24(m,4H).

[0253] 13 C NMR(101MHz,DMSO-d6)δ169.56,162.81,161.43,159.50,157.76,149.89,112.41,10 4.10,54.15,54.03,45.86,39.00,32.69,29.32,28.72,26.64,25.52,23.69,13.93.

[0254] Test Example: Inhibitory activity of compounds based on the 6-methylfurano[2,3-d]pyrimidine-4(3H)-one structure against human non-small cell lung cancer cells H460 and H460 DNMT3A-HDAC6 double knockout cells.

[0255] H460 DNMT3AHDAC6 double knockout cells were constructed using CRISPR-Cas9 technology. RPMI 1640 medium was used as the basal medium, and a cell culture medium containing 10% fetal bovine serum was prepared. Cells were cultured in a 37°C incubator containing 5% CO2. The culture medium was changed daily. When the cells reached 80%–90% confluence, the original culture medium was discarded, the cells were washed once with PBS, digested with 0.25% trypsin, and passaged according to experimental requirements.

[0256] Weigh a portion of the compound powder prepared in Examples 1 to 31 using an analytical balance. Calculate the required volume of DMSO based on the molecular weight and the mass of the compound, and prepare a stock solution of the same concentration (100 mM / L). Use a vortex mixer to shake and fully dissolve and mix the solution. Dispense the solution into 200 μL centrifuge tubes, avoid repeated freeze-thaw cycles, and store at -40°C. Prepare the working solution fresh each time.

[0257] Cells in good growth condition during the logarithmic growth phase were digested with 0.25% trypsin, then dispersed into a single-cell suspension by pipetting with culture medium. After counting, the cells were seeded at an appropriate density (H460 Vector, H460DNMT3AHDAC6-KO2500 cells / well) into 96-well plates at 100 μL / well and cultured in a 37°C incubator with 5% CO2.

[0258] Drug screening: After the cells were incubated overnight, cells in good growth condition during the logarithmic growth phase were taken, dispersed into single-cell suspensions, and plated at a density of 2500 cells / well (H460 Vector, H460 DNMT3AHDAC6-KO). After 18 hours of plated cells, 31 test compounds at a concentration of 10 μM were added to the two types of cells, with six replicates per compound, and incubated for 96 hours.

[0259] Drug re-screening: After overnight cell culture, five different concentration gradients of the drug were prepared according to experimental requirements. Each group included six replicates, and each plate also included a control group. During the experiment, the drug was diluted to concentration gradients of 100 μM, 10 μM, 1 μM, 0.1 μM, and 0.01 μM and added, with a continuous treatment time of 96 hours. After drug addition, the cells were cultured for the corresponding time in a 5% CO2 incubator at 37°C, and the cell status was observed using an inverted microscope.

[0260] After the drug treatment period, 10 μL of 2.5 mg / mL MTT solution was added to each well, and the cells were incubated at 37°C for another 4 hours. Then, the supernatant was removed, and 100 μL of dimethyl sulfoxide was added to each well, followed by shaking at an appropriate speed for 5 minutes to dissolve the purple crystals. The 96-well plate was then placed in a microplate reader, and the optical density at 492 nm was measured to calculate the effect of the drug on cell proliferation.

[0261] Calculate the cell proliferation rate, inhibition rate, and drug resistance index for each group.

[0262] Cell viability % = (OD value of drug-treated group / OD value of blank group) × 100%;

[0263] Inhibition rate % = (1 - OD value of the treated group / OD value of the blank group) × 100%;

[0264] GraphPadPrism 8.0 software was used to plot the drug's effective concentration and its inhibition rate on cell growth. Then, SPSS 20.0 was used to calculate the half-maximal inhibitory concentration (IC50) using the regression-Probit method. 50 )value.

[0265] The inhibition rates of the compounds of this invention against H460 Vector and H460 DNMT3A-HDAC6 double knockout cells are shown in Table 1.

[0266] Table 1. Inhibition rate (%) of the compounds of this invention

[0267]

[0268]

[0269] Furthermore, FQX-1B, FQX-3C, YQ-1C, FQX-1D, FQX-2B, YQ-1B, and YQ-2A were diluted to concentration gradients of 100 μM, 10 μM, 1 μM, 0.1 μM, and 0.01 μM, respectively, for inhibition rate testing, and IC50 was calculated. 50 The values ​​are shown in Table 2.

[0270] Table 2 IC of the compounds of the present invention 50 Value (μM)

[0271]

[0272] As shown in Tables 1 and 2, compounds FQX-1B, FQX-3C, YQ-1C, FQX-1D, and FQX-2D significantly inhibited the H460 Vector cell line of human non-small cell lung cancer cells more than H460-DNMT3A / HDAC6 double knockout cells. Compounds FQX-1B, FQX-2B, YQ-1B, YQ-2A, YQ-7A, and FQX-1D all showed strong inhibition rates against both H460 Vector and H460-DNMT3A / HDAC6 double knockout cells. Compound FQX-1D exhibited stronger selective inhibitory effects against DNMT3A and HDAC6.

[0273] To determine the interactions of the compounds of this invention with DNMT3A and HDAC6, cell heat transfer assay (CETSA) was performed on compound FQX-1D. The results are as follows: Figure 1 As shown. According to Figure 1 It can be seen that the thermal stability of DNMT3A and HDAC6 was significantly enhanced after treatment with compound FQX-1D compared with the control group, indicating that there is a direct interaction between compound FQX-1D and intracellular DNMT3A and HDAC6.

[0274] The effect of compound FQX-1D on the overall methylation status of genomic DNA in H460 vector cells was assessed using methylation dot blot analysis. The results are as follows: Figure 2 As shown. According to Figure 2 It can be seen that cells treated with the DNMT inhibitor 5-Aza or the compound FQX-1D exhibit a significant concentration-dependent reduction in global methylation.

[0275] Acetylated α-tubulin (Ac-α-tubulin) is a major cytoplasmic substrate of HDAC6 and has a significant impact on tumorigenesis. It is commonly used as an indicator of HDAC6 deacetylase activity. The expression level of Ac-α-tubulin in H460 cells was detected, and the results are as follows: Figure 3 As shown. According to Figure 3 As can be seen, consistent with the HDAC6 inhibitor WT-161, compound FQX-1D significantly increased Ac-α-tubulin levels in a dose-dependent manner by inhibiting HDAC6 activity, with better effects than WT-161 as a positive control at high doses.

[0276] Enzyme activity of compound FQX-1D was detected, and the results are as follows: Figure 4 As shown. According to Figure 4 It can be seen that compound FQX-1D exhibits concentration-dependent inhibitory effects on both DNMT and HDAC6 enzymes, with an IC50 value of [missing value]. 50 The value is close to the IC50 of the positive drug. 50The results above indicate that compound FQX-1D has a strong binding affinity for DNMT3A and HDAC6, effectively inhibiting their respective activities.

[0277] The effects of compound FQX-1D on the proliferation, migration, and self-renewal abilities of H460 carrier cells were tested and compared with DNMT3A-HDAC6 double knockout (Double KO) cells. Results showed that the proliferation and migration abilities of Double KO cells were partially impaired compared to carrier cells, indicating the roles of DNMT3A and HDAC6 in tumorigenesis. Colony formation and spheroid formation assays showed that compound FQX-1D significantly inhibited the formation of colonies and the number of tumor spheroids by carrier cells in a concentration-dependent manner, but had no significant inhibitory effect on Double KO cells. Similarly, transpore migration assays confirmed that compound FQX-1D inhibited the migration ability of carrier cells in a concentration-dependent manner, while having little inhibitory effect on Double KO cells.

[0278] In a subcutaneous xenograft model established using H460 vector and DNMT3A / HDAC6 double knockout cells, the targeting activity of compound FQX-1D in vivo was investigated. Results showed that, consistent with in vitro experiments, knockout of DNMT3A and HDAC6 in mice resulted in a reduction of tumor volume by more than 60% compared to the Vector group. Treatment with compound FQX-1D (20 mg / kg) induced tumor regression in mice with H460 vector cell xenografts, while also exhibiting minimal inhibitory effect on the growth of double-KO cell-derived tumors. The endpoint tumor weight inhibition rates were 67.83% and 27.48%, respectively. These results demonstrate that compound FQX-1D can inhibit the proliferation of human non-small cell lung cancer cells in vivo by specifically inhibiting DNMT3A and HDAC6.

[0279] As can be seen from the above examples, the compound provided by the present invention exhibits selectivity for DNMT3A and HDAC6, and has strong inhibitory effects on both DNMT3A and HDAC6. Its inhibitory activity on human non-small cell lung cancer cells H460 is much higher than that on H460 DNMT3A-HDAC6 double knockout cells, and it has application value as a drug therapeutic agent for treating diseases and disorders related to DNMT3A and HDAC6.

[0280] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. Other embodiments can be obtained based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A compound based on the structure of 6-methylfurano[2,3-d]pyrimidine-4(3H)-one, characterized in that, It has the structure shown in YQ-7A, FQX-1B, FQX-2B, YQ-7B, YQ-1B, YQ-2A, FQX-2A, or FQX-1D: ； ； 。 2. A compound based on the structure of 6-methylfurano[2,3-d]pyrimidine-4(3H)-one, characterized in that, It has the structure shown in WZ-1B: 。 3. The method for preparing the compound based on the 6-methylfurano[2,3-d]pyrimidine-4(3H)-one structure according to claim 1, characterized in that, Includes the following steps: (1) Ethyl 2-chloro-3-oxobutyrate was mixed with malononitrile and subjected to a cyclization reaction to obtain a compound with the structure shown in Formula II; (2) The compound with the structure shown in Formula II is mixed with formic acid and acetic anhydride to carry out a cyclization reaction to obtain the compound with the structure shown in Formula III; (3) The compound with the structure shown in Formula III is mixed with a substituted alkane, a first basic compound and a first organic solvent to carry out a substitution reaction to obtain the compound with the structure shown in Formula IV; (4) The compound with the structure shown in Formula IV is mixed with a substituted amine, a second basic compound, and a second organic solvent to carry out a substitution reaction to obtain the compound with the structure shown in Formula V; The substituted amine has the structural formula shown in Formula VIII: RH Formula VIII; (5) The compound with the structure shown in Formula V is mixed with a third basic compound and a third solvent and subjected to a hydrolysis reaction to obtain the compound with the structure shown in Formula VI; (6) The compound with the structure shown in Formula VI is mixed with an amino-substituted carboxylic acid ester, a condensing agent and a fourth organic solvent to carry out a condensation reaction to obtain the compound with the structure shown in Formula VII; (7) The compound with the structure shown in Formula VII is mixed with hydroxylamine, water, a fourth basic compound and a fifth organic solvent to carry out a hydroxylamine reaction to obtain the compound based on the 6-methylfurano[2,3-d]pyrimidine-4(3H)-one structure; ； The substituted alkane is a dibromo-substituted alkane or a bromochloro-substituted alkane; The dibromosubstituted alkane is 1,2-dibromoethane or 1,3-dibromopropane; The bromochlorosubstituted alkane is 1-bromo-2-chloroethane or 1-bromo-3-chloropropane; The amino-substituted carboxylic acid ester is methyl p-aminobenzoate, methyl 5-aminopyridine-2-carboxylate, or methyl 7-aminoheptanoate. The R, n, and L mentioned above all correspond to the corresponding structures in claim 1.

4. The preparation method according to claim 3, characterized in that, The molar ratio of the compound with the structure shown in Formula II to formic acid is 1:10~30; The molar ratio of the compound with the structure shown in Formula II to acetic anhydride is 1:5~10.

5. The preparation method according to claim 3, characterized in that, The molar ratio of the compound with the structure shown in Formula III to the substituted alkane is 1:3 to 10; The molar ratio of the compound with the structure shown in Formula III to the first basic compound is 1:1.2~5; The mass-to-volume ratio of the compound with the structure shown in Formula III to the first organic solvent is 1 g: (5~30) mL.

6. The preparation method according to claim 3, characterized in that, The molar ratio of the compound with the structure shown in Formula VI to the amino-substituted carboxylic acid ester is 1:0.8~1.2; The molar ratio of the compound with the structure shown in Formula VI to the condensing agent is 1:1~2; The mass-to-volume ratio of the compound with the structure shown in Formula VI to the fourth organic solvent is 1 g: (5~50) mL.

7. The preparation method according to claim 3, characterized in that, The mass ratio of hydroxylamine to water is 1:1 to 10; The molar ratio of the compound with the structure shown in Formula VII to hydroxylamine is 1:20~100; The molar ratio of the compound with the structure shown in Formula VII to the fourth basic compound is 1:5~20; The mass-to-volume ratio of the compound with the structure shown in Formula VII to the fifth organic solvent is 1 g: (5~50) mL.

8. The use of the compound based on the 6-methylfurano[2,3-d]pyrimidine-4(3H)-one structure as described in claim 1 or 2, or the compound based on the 6-methylfurano[2,3-d]pyrimidine-4(3H)-one structure obtained by the preparation method described in any one of claims 3 to 7, in the preparation of a medicament for treating diseases related to DNA methyltransferase 3A or histone deacetylase.

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