Dihydroquinazolinone-containing n-hydroxy amide compounds, methods of making and using the same

Abstract

The application discloses a dihydroquinazolinone-containing N-hydroxy amide compound and a preparation method and application thereof, relates to the technical field of medicinal chemistry, and utilizes the splicing principle to design and synthesize dihydroquinazolinone-containing N-hydroxy amide compounds with novel structures, and the compounds are tested in terms of HDAC6 protein affinity and anti-proliferation activity of cervical cancer cell Siha, and the results show that the compounds E1-E5 have better affinity to the HDAC6 protein, the IC 50 values are 4.1-85.4 nM, wherein the affinity of the compound E4 to the HDAC6 protein is higher than that of a positive control SAHA; and the compounds E1-E5 can better inhibit the proliferation of the cervical cancer cell Siha, the IC 50 values are 2.4-9.3 muM, wherein the activity of the compound E4 is the strongest.

Description

Technical Field

[0001] This invention relates to the field of medicinal chemistry, specifically to an N-hydroxyamide compound containing dihydroquinazolinone, its preparation method, and its application. Background Technology

[0002] Histone acetylation and deacetylation mainly occur at the α-amino terminus of lysine residues, catalyzed by both histone acetyltransferases and histone deacetylases (HDACs). Abnormally high expression of HDACs can lead to various diseases, such as cancer, inflammation, autoimmune diseases, and other neurological disorders.

[0003] Eighteen HDACs have been identified in the human genome, which can be divided into four classes: Class I (HDACs 1, 2, 3, and 8), Class II (IIa, 4, 5, 7, and 9), Class IIb (HDACs 6 and 10), and Class IV (HDAC11). HDAC6 is the most potent member of the HDAC family, containing 1216 amino acid residues and possessing a nuclear export signaling domain, two catalytic domains, a fourteen-peptide structure, and a zinc finger ubiquitin-binding domain unique to HDAC6. HDAC6 is highly expressed in various cancer cells, such as breast cancer, liver cancer, cervical cancer, and melanoma. Therefore, HDAC6 protein has attracted attention as a target for anticancer drugs.

[0004] Zn 2+ HDAC6-dependent inhibitors consist of a surface recognition cap, a zinc ion-binding group, and a linker group. The activity of HDAC6 inhibitors can be enhanced by altering or modifying the structure of the surface recognition cap and linker group. Hydroxyxamic acid is a common zinc-containing compound used in HDAC6 inhibitors. 2+ Chelating groups, such as vorinostatin (SAHA), have a good therapeutic effect on cutaneous T-cell lymphoma; trichostatin A, as an inhibitor of HDAC1, HDAC4 and HDAC6, has a strong inhibitory effect on breast cancer cells; and isoxazole-3-isohydroxyoxime ester (SS-208), an HDAC6 inhibitor, has a good inhibitory effect on melanoma.

[0005] Although there are many HDAC6 inhibitors under development, marketed HDAC inhibitors generally suffer from poor biological activity and limited efficacy against solid tumors. Therefore, the development of novel HDAC6 inhibitors holds great promise for future applications. Summary of the Invention

[0006] The technical problem to be solved by the present invention is to provide an N-hydroxyamide compound containing dihydroquinazolinone and a method for preparing the same. This compound can be used not only to prepare histone deacetylase 6 inhibitors, but also to prepare drugs for the prevention and / or treatment of diseases related to abnormal expression of histone deacetylase 6 activity.

[0007] The technical problem to be solved by this invention is achieved by the following technical solution:

[0008] One objective of this invention is to provide an N-hydroxyamide compound containing dihydroquinazolinone, the structural formula of which is as follows:

[0009]

[0010] In Formula I, R is at least one of mono- or poly-substituted halogens, alkyl groups, and alkoxy groups.

[0011] In some specific implementations, R is 4-F, 4-CH3, 4-CH3O, 3,4-CH3O, or 3,5-CH3O.

[0012] A second objective of this invention is to provide a method for preparing the aforementioned N-hydroxyamide compound containing dihydroquinazolinone, comprising the following steps:

[0013] (1) 2-Aminobenzamide undergoes a substitution reaction with methyl 5-bromopentanoate to give intermediate A;

[0014] (2) Intermediate A undergoes a cyclization reaction with substituted benzaldehyde to give intermediate B;

[0015] (3) Intermediate B undergoes a hydrolysis reaction to obtain intermediate C;

[0016] (4) Intermediate C undergoes an amide condensation reaction with O-benzyloxyhydroxylamine hydrochloride to obtain intermediate D;

[0017] (5) Intermediate D undergoes a hydrogenation reaction to obtain compound I.

[0018] The synthesis route is as follows:

[0019]

[0020] In a further technical solution, the substitution reaction is carried out under the action of an acid-binding agent. Preferably, the acid-binding agent is at least one selected from triethylamine, pyridine, N,N-diisopropylethylamine, sodium carbonate, and potassium carbonate.

[0021] In a further technical solution, the molar ratio of 2-aminobenzamide to methyl 5-bromopentanoate and the acid-binding agent is 1:(1-1.1):(2-3).

[0022] In a further technical solution, the ring-closing reaction is carried out in the presence of a catalyst. Preferably, the catalyst is at least one selected from p-toluenesulfonic acid and benzenesulfonic acid.

[0023] In a further technical solution, the molar ratio of intermediate A to substituted benzaldehyde is 1:(1-1.1).

[0024] In a further technical solution, the hydrolysis reaction is carried out under alkaline conditions, wherein the alkalinity is provided by at least one of sodium carbonate, potassium carbonate, sodium hydroxide, and potassium hydroxide.

[0025] In a further technical solution, the amide condensation reaction is carried out in the presence of a condensing agent. Preferably, the condensing agent is at least one selected from 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCl), carbonyl diimidazole (CDI), O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethylurea hexafluorophosphate (HATU), benzotriazole-N,N,N′,N′-tetramethylurea hexafluorophosphate (HBTU), and 1-n-propylphosphoric anhydride (T3P).

[0026] In a further technical solution, the molar ratio of intermediate C to O-benzyloxyhydroxylamine hydrochloride is 1:(1-1.1).

[0027] In a further technical solution, the hydrogenation reaction is carried out under the action of a catalyst. Preferably, the catalyst is at least one selected from palladium on carbon, platinum on carbon, and Raney nickel, and the hydrogen pressure is 30-50 Psi.

[0028] A third objective of this invention is to provide the application of the aforementioned dihydroquinazolinone-containing N-hydroxyamide compounds in the preparation of histone deacetylase 6 inhibitors.

[0029] A fourth objective of this invention is to provide the use of the aforementioned dihydroquinazolinone-containing N-hydroxyamide compounds in the preparation of medicaments for the prevention and / or treatment of diseases associated with abnormal expression of histone deacetylase 6 activity.

[0030] The diseases associated with abnormal expression of histone deacetylase 6 activity include malignant tumors, inflammation, neurodegenerative diseases, renal fibrosis, and autoimmune diseases.

[0031] The beneficial effects of this invention are as follows: This invention utilizes the principle of combination to design and synthesize novel N-hydroxyamide compounds containing dihydroquinazolinone. These compounds were then tested for their affinity for HDAC6 protein and their antiproliferative activity against cervical cancer cells (Siha). The results showed that compounds E1-E5 exhibited good affinity for HDAC6 protein, with an IC50 concentration of [missing value]. 50The values ​​ranged from 4.1 to 85.4 nM, with compound E4 showing a higher affinity for HDAC6 protein than the positive control SAHA; and compounds E1-E5 all effectively inhibited the proliferation of cervical cancer cells Siha, with IC50 values ​​of 4.1-85.4 nM. 50 The values ​​ranged from 2.4 to 9.3 μM, with compound E4 exhibiting the strongest activity. Attached Figure Description

[0032] Figure 1 For intermediate A 1 H NMR spectrum;

[0033] Figure 2 For intermediate B1 1 H NMR spectrum;

[0034] Figure 3 For intermediate C1 1 H NMR spectrum;

[0035] Figure 4 For intermediate D1 1 H NMR spectrum;

[0036] Figure 5 For compound E1 1 H NMR spectrum;

[0037] Figure 6 For compound E1 13 C NMR spectrum. Detailed Implementation

[0038] To make the technical means, creative features, objectives and effects of this invention easier to understand, the invention will be further described below with reference to specific embodiments and illustrations.

[0039] Example 1: Synthesis of Intermediate A

[0040]

[0041] 2-Aminobenzamide (2.72 g, 20.0 mmol), methyl 5-bromopentanoate (4.30 g, 22 mmol), and triethylamine (6.06 g, 60 mmol) were added to 80 mL of toluene and heated to 95 °C for 6 h. The reaction mixture was cooled to room temperature, washed twice with water, and concentrated under reduced pressure to remove the solvent, yielding intermediate A, a pale yellow solid in 92% yield. 1H NMR (600MHz, DMSO-d6) δ8.11(s,1H),7.80(s,1H),7.59(dd,J=7.9,1.5Hz,1H),7.25(ddd,J=8.5,7.2,1.5Hz,1H),7.11(s ,1H),6.65(d,J=8.0Hz,1H),6.53-6.48(m,1H),3.58(s,3H),3.09(t,J=6.7Hz,2H),2.38-2.32(m,2H),1.65-1.54(m,4H).

[0042] Example 2: Synthesis of intermediates B1-B5

[0043] Synthesis of intermediate B1:

[0044] Intermediate A (1.00 g, 4.0 mmol), 4-fluorobenzaldehyde (0.546 g, 4.4 mmol), p-toluenesulfonic acid (0.069 g, 0.4 mmol), and 40 mL of anhydrous ethanol were reacted at 78 °C for 8 h under nitrogen protection. The reaction solution was cooled to room temperature, and the ethanol was removed by concentration under reduced pressure. The residue was dissolved in dichloromethane, and then washed successively with water, saturated sodium carbonate aqueous solution, and water. The dichloromethane was removed by concentration under reduced pressure to give 1.07 g of intermediate B1. It was an off-white solid with a yield of 75.0%. 1 H NMR (600MHz, DMSO-d6) δ8.86(d,J=4.0Hz,1H),7.87(d,J=7.6Hz,1H),7.57(dd,J =8.2,7.3Hz,1H),7.52(dd,J=8.3,5.7Hz,2H),7.35(t,J=8.3Hz,2H),7.04(d,J= 8.4Hz,1H),6.93(t,J=7.4Hz,1H),5.94(d,J=3.9Hz,1H),3.73(dd,J=14.4,6.2H z,1H),3.54(s,3H),3.38-3.28(m,1H),2.50(t,J=7.0Hz,2H),1.81-1.67(m,4H).

[0045]

[0046] Synthesis of intermediate B2:

[0047] The synthesis method of intermediate B2 is the same as that of intermediate B1, except that 4-fluorobenzaldehyde is replaced with 4-methylbenzaldehyde.

[0048]

[0049] Synthesis of intermediate B3:

[0050] The synthesis method of intermediate B3 is the same as that of intermediate B1, except that 4-fluorobenzaldehyde is replaced with 4-methoxybenzaldehyde.

[0051]

[0052] Synthesis of intermediate B4:

[0053] The synthesis method of intermediate B4 is the same as that of intermediate B1, except that 4-fluorobenzaldehyde is replaced with 3,4-dimethoxybenzaldehyde.

[0054]

[0055] Synthesis of intermediate B5:

[0056] The synthesis method of intermediate B5 is the same as that of intermediate B1, except that 4-fluorobenzaldehyde is replaced with 3,5-dimethoxybenzaldehyde.

[0057]

[0058] Example 3: Synthesis of intermediates C1-C5

[0059] Synthesis of intermediate C1:

[0060] Intermediate B1 (1.07 g, 3.0 mmol) and anhydrous sodium carbonate (0.954 g, 9.0 mmol) were added to 40 mL of a mixed solution of distilled water and methanol (1:1 volume ratio). The mixture was heated to 60 °C for 6 h under nitrogen protection. The reaction solution was cooled to room temperature and extracted with 20 mL of methyl tert-butyl ether to remove some impurities. The aqueous layer was adjusted to pH 4-5 with dilute hydrochloric acid, and then extracted twice with 40 mL of ethyl acetate. The combined ethyl acetate layers were washed with water and concentrated under reduced pressure to remove the ethyl acetate, yielding 0.823 g of intermediate C1. The solid was off-white, with a yield of 80.2%. 1 H NMR (600MHz, DMSO-d6) δ12.03(s,1H),8.66(d,J=3.5Hz,1H),7.67(d,J=7.6Hz,1H ),7.36(dd,J=8.2,7.3Hz,1H),7.32(dd,J=8.4,5.6Hz,2H),7.13(dd,J=12.6,5.0 Hz,2H),6.84(d,J=8.4Hz,1H),6.72(t,J=7.4Hz,1H),5.80-5.66(m,1H),3.59-3. 51(m,1H),3.12(dt,J=14.4,7.1Hz,1H),2.22(t,J=7.1Hz,2H),1.65-1.45(m,4H).

[0061]

[0062] Synthesis of intermediate C2:

[0063] The synthesis method of intermediate C2 is the same as that of intermediate C1, except that intermediate B1 is replaced by intermediate B2.

[0064]

[0065] Synthesis of intermediate C3:

[0066] The synthesis method of intermediate C3 is the same as that of intermediate C1, except that intermediate B1 is replaced by intermediate B3.

[0067]

[0068] Synthesis of intermediate C4:

[0069] The synthesis method of intermediate C4 is the same as that of intermediate C1, except that intermediate B1 is replaced by intermediate B4.

[0070]

[0071] Synthesis of intermediate C5:

[0072] The synthesis method of intermediate C5 is the same as that of intermediate C1, except that intermediate B1 is replaced by intermediate B5.

[0073]

[0074] Example 4: Synthesis of intermediates D1-D5:

[0075] Synthesis of intermediate D1:

[0076] Intermediate C1 (0.684 g, 2 mmol), O-benzyloxyhydroxylamine hydrochloride (0.271 g, 1.1 mmol), EDCl (0.475 g, 2.4 mmol), HOBt (0.019 g, 0.14 mmol), and triethylamine (0.73 g, 7.2 mmol) were added to 20 mL of dichloromethane and reacted at room temperature for 10 h. The reaction solution was washed successively with water, saturated sodium carbonate aqueous solution, and 1 mol / L dilute hydrochloric acid. The solvent was removed by concentration under reduced pressure. The residue was purified by column chromatography (eluent: dichloromethane / methanol) to give intermediate D1, an off-white solid in 78.9% yield. 1H NMR (600MHz, DMSO-d6) δ10.93(s,1H),8.66(d,J=3.8Hz,1H),7.66(dd,J=7.7,1.6Hz,1H ),7.41-7.32(m,6H),7.31(dd,J=8.7,5.5Hz,2H),7.14(dd,J=12.4,5.4Hz,2H),6.83(d ,J=8.4Hz,1H),6.72(t,J=7.4Hz,1H),5.72(d,J=4.0Hz,1H),4.75(s,2H),3.53(dd,J=1 4.0, 8.4Hz, 1H), 3.10 (dd, J = 15.3, 7.1Hz, 1H), 1.96 (t, J = 6.6Hz, 2H), 1.59-1.44 (m, 4H).

[0077]

[0078] Synthesis of intermediate D2:

[0079] The synthesis method of intermediate D2 is the same as that of intermediate D1, except that intermediate C1 is replaced by intermediate C2.

[0080]

[0081] Synthesis of intermediate D3:

[0082] The synthesis method of intermediate D3 is the same as that of intermediate D1, except that intermediate C1 is replaced by intermediate C3.

[0083]

[0084] Synthesis of intermediate D4:

[0085] The synthesis method of intermediate D4 is the same as that of intermediate D1, except that intermediate C1 is replaced by intermediate C4.

[0086]

[0087] Synthesis of intermediate D5:

[0088] The synthesis method of intermediate D5 is the same as that of intermediate D1, except that intermediate C1 is replaced by intermediate C5.

[0089]

[0090] Example 5: Synthesis of compounds E1-E5

[0091] Synthesis of compound E1:

[0092] Intermediate D1 (0.447 g, 1 mmol) and palladium on carbon (30 mg) were added to a hydrogenation flask, followed by the addition of 10 mL of methanol. The mixture was purged three times with nitrogen and three times with hydrogen. The reaction was then carried out at 30 °C for 8 h under a hydrogen pressure of 30 Psi. Palladium on carbon was removed by filtration, and methanol was removed by concentration under reduced pressure. The residue was purified by column chromatography (eluent: dichloromethane / methanol) to give compound E1. 0.24 g of a yellowish-brown solid was obtained, with a yield of 67.2%. 1 H NMR (600MHz, DMSO-d6) δ10.36(s,1H),8.67(d,J=3.7Hz,2H),7.66(dd,J=7.6,1.4H z,1H),7.40-7.34(m,1H),7.31(dd,J=8.5,5.5Hz,2H),7.14(t,J=8.8Hz,2H),6.83 (d,J=8.4Hz,1H),6.72(t,J=7.4Hz,1H),5.72(d,J=3.8Hz,1H),3.55(dd,J=14.2,8 .5Hz,1H),3.11(dd,J=14.6,6.5Hz,1H),1.96(t,J=6.9Hz,2H),1.60-1.43(m,4H). 13 C NMR (151MHz, DSMO-d6) δ1168.88,162.54,161.80 (J=244.3Hz),146.19,138.13,138.11,133.87,128.2 1(J=8.4Hz),127.88,116.90,116.34,115.29(J=21.5Hz),112.70,70.03,48.11,32.01,26.52,22.52. [M+H] + 358.16.

[0093]

[0094] Synthesis of compound E2:

[0095] The synthesis method for compound E2 is the same as that for compound E1, except that intermediate D1 is replaced by intermediate D2. It is a yellowish-brown solid with a yield of 71.2%. 1H NMR (600MHz, DMSO-d6) δ10.37(s,1H),8.69(s,1H),8.61(s,1H),7.65(d,J=7.1Hz,1 H),7.35(t,J=7.5Hz,1H),7.15(d,J=7.9Hz,2H),7.10(d,J=7.9Hz,2H),6.80(d,J=8. 3Hz,1H),6.70(t,J=7.3Hz,1H),5.65(d,J=2.9Hz,1H),3.52(dd,J=13.8,8.2Hz,1H), 3.08(dd,J=14.4,6.5Hz,1H),2.23(s,3H),1.96(t,J=6.5Hz,2H),1.55-1.47(m,4H). 13 C NMR (151MHz, DSMO-d6) δ168.78,162.62,146.36,138.80,37.34,133.74,128.97,1 27.81,126.07,116.66,116.38,112.47,70.53,47.98,31.98,26.46,22.54,20.64. [M+H] + 354.18.

[0096]

[0097] Synthesis of compound E3:

[0098] The synthesis method for compound E3 is the same as that for compound E1, except that intermediate D1 is replaced by intermediate D3. It is a yellowish-brown solid with a yield of 74.6%. 1 H NMR (600MHz, DMSO-d6) δ10.37(s,1H),8.69(s,1H),8.61(s,1H),7.63(d,J=7. 2Hz,1H),7.43(d,J=7.6Hz,2H),7.34(t,J=7.4Hz,1H),6.93(d,J=7.9Hz,2H), 6.85(d,J=8.3Hz,1H),6.65-6.57(m,1H),5.64(d,J=3.2Hz,1H),3.62(s,3H), 3.56-3.51m,1H),3.15-3.09(m,1H),1.96(t,J=6.4Hz,2H),1.59-1.51(m,4H). 13C NMR (151MHz, DSMO-d6) δ168.82,162.64,157.41,148.45,133.87,128.61,127.80,1 26.07,118.32,116.87,116.38,115.31,70.53,56.36,47.98,31.98,26.59,22.57. [M+H] + 370.18.

[0099]

[0100] Synthesis of compound E4:

[0101] The synthesis method for compound E4 is the same as that for compound E1, except that intermediate D1 is replaced by intermediate D4. It is a yellowish-brown solid with a yield of 74.6%. 1 H NMR (600MHz, DMSO-d6) δ10.36(s,1H),8.69(s,1H),8.56(s,1H),7.66(d,J=6.2Hz,1H),7.36(s,1H),6.93(s,1H),6.88-6.77(m,2H),6. 76-6.64(m,2H),5.63(s,1H),3.69(s,3H),3.57(s,3H),3.54-3.47(m,1H),3.10-3.04(m,1H),1.96(t,J=6.5Hz,2H),1.61-1.45(m,4H). 13 C NMR (151MHz, DSMO-d6) δ168.84,162.66,148.60,146.55,133.87,133.75,127.78,118.25, 116.68,116.42,112.41,111.42,110.21,70.61,55.48,55.39,47.87,32.01,26.38,22.58. [M+H] + : 400.19.

[0102]

[0103] Synthesis of compound E5:

[0104] The synthesis method for compound E5 is the same as that for compound E1, except that intermediate D1 is replaced by intermediate D5. It is a yellowish-brown solid with a yield of 79.3%. 1H NMR(600MHz,DMSO-d6)δ10.35(s,1H),8.68(s,1H),8.62(s,1H),7.64(d,J=7 .4Hz,1H),7.37(t,J=7.5Hz,1H),6.84(d,J=8.3Hz,1H),6.71(t,J=7.4Hz,1H ),6.40(s,3H),5.61(d,J=3.3Hz,1H),3.66(s,6H),3.56(dd,J=13.7,8.0Hz, 1H), 3.10 (dd, J=14.5, 6.8Hz, 1H), 1.96 (t, J=6.4Hz, 2H), 1.61-1.46 (m, 4H). 13 C NMR (151MHz, DSMO-d6) δ168.87,162.58,160.44,146.42,144.16,133.80,127.81, 116.75,116.40,112.37,104.49,98.98,70.57,55.11,48.18,32.01,26.61,22.54. [M+H] + : 400.19.

[0105]

[0106] Example 6: Assay of the inhibitory activity of compounds E1-E5 against HDAC6 protein

[0107] The inhibitory activity of compounds E1-E5 against HDAC6 protein was detected using the HDAC6 Fluorogenic Assay Kit (BPS Bioscience, USA). The specific steps are as follows:

[0108] 1) Prepare a 0.1% BSA (Solepro) solution;

[0109] 2) Dilute the DMSO solution of Trichostatin A from 200 μM to 20 μM using the HDAC buffer solution provided in the kit;

[0110] 3) Dilute the Fluorogenic HDAC substrate in the kit from 5 mM to 200 μM using HDAC buffer solution;

[0111] 4) Prepare a 7 ng / μL solution of the HDAC6 recombinant human protein from the kit using HDAC buffer solution;

[0112] 5) Add 5 μL of Fluorogenic HDAC substrate (200 μM), 5 μL of BSA solution (0.1%), and 30 μL of HDAC detection buffer to a black 96-well plate;

[0113] 6) Add 5 μL of aqueous solution containing 10% DMSO to the control group and blank group, add 5 μL of test sample of different concentrations to the test group, and add DMSO solution of Trichostatin A (20 μM) to the positive control group.

[0114] 7) Add 5 μL of HDAC detection buffer to the blank control group;

[0115] 8) Except for the blank group, add HDAC6 human recombinant enzyme (7ng / μL) to each group and then incubate at 37℃ for 30 min;

[0116] 9) Add 50 μL of HDAC Developer solution from the kit to each well, and then incubate at room temperature for 15 min;

[0117] 10) Measure absorbance in a fluorescence detector that excites light in the wavelength range of 350-380 nm and detects the emitted light in the wavelength range of 440-460 nm;

[0118] Table 1. In vitro inhibitory activity of compounds E1-E5 against HDAC6.

[0119] E1 E2 E3 E4 E5 SAHA <![CDATA[IC 50 (nM)]]> 85.4±7.1 35.5±5.2 26.1±4.4 4.1±0.6 21.1±1.6 13.1±0.8

[0120] SAHA (Vorinostat) served as the positive control.

[0121] As shown in Table 1, compounds E1-E5 yield IC 50 The values ​​range from 4.1 to 85.4 nM, indicating that they all have good suppression effects on HDAC6, with compound E4 showing the highest IC50 value. 50 The value was 4.1 ± 0.6 nM, which was lower than the positive control SAHA's value of 13.1 ± 0.8 nM.

[0122] Example 7: CCK8 assay to determine the cytotoxicity of compounds E1-E5 against cervical cancer cells Siha.

[0123] Siha cells were cultured in RPMI-R1640 medium (Beyotime) containing 10% fetal bovine serum and 1% penicillin-streptomycin solution in a cell culture incubator at 37°C with 5% CO2 air. Siha cells in the logarithmic growth phase were collected, and approximately 10 μL of each cell was added to each well of a 96-well plate. 4Cells were incubated in an incubator for 12 hours, then different concentrations of compounds E1-E5 were added, and incubation continued for 48 hours. Then, 10 μL of CCK8 solution (10 mg / mL, Adamas Life) was added, and incubation continued for 2 hours. The absorbance of each well was measured at 450 nm using a microplate reader, and the IC50 was calculated using Prism 8.0 software. 50 value.

[0124] Table 2. In vitro inhibitory activity of compounds E1-E5 against cervical cancer cells Siha.

[0125] E1 E2 E3 E4 E5 <![CDATA[IC 50 (μM)]]> 4.5±0.2 9.3±0.5 6.1±0.5 2.4±0.2 8.1±0.4

[0126] Table 2 shows that compounds E1-E5 have a strong inhibitory effect on cervical cancer cells Siha, IC50. 50 The activity ranged from 2.4 to 9.3 μM, with compound E4 exhibiting the strongest activity (IC50). 50 The value is 2.4 ± 0.2 μM.

[0127] The foregoing has shown and described the basic principles, main features, and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The embodiments and descriptions in the specification are merely illustrative of the principles of the invention. Various changes and modifications can be made to the invention without departing from its spirit and scope, and all such changes and modifications fall within the scope of the present invention as claimed. The scope of protection of this invention is defined by the appended claims and their equivalents.

Claims

1. An N-hydroxyamide compound containing dihydroquinazolinone, the structure of which is shown in Formula I: ； In Formula I, R is 4-F, 4-CH3, 4-CH3O, 3,4-dimethoxy, or 3,5-dimethoxy.

2. The method for preparing the N-hydroxyamide compound containing dihydroquinazolinone according to claim 1, characterized in that, Includes the following steps: (1) 2-Aminobenzamide undergoes a substitution reaction with methyl 5-bromopentanoate to give intermediate A; (2) Intermediate A undergoes a cyclization reaction with substituted benzaldehyde to give intermediate B; (3) Intermediate B undergoes a hydrolysis reaction to obtain intermediate C; (4) Intermediate C undergoes an amide condensation reaction with O-benzylhydroxylamine hydrochloride to give intermediate D; (5) Intermediate D undergoes a hydrogenation reaction to obtain compound I; The synthesis route is as follows: 。 3. The preparation method according to claim 2, characterized in that: The substitution reaction is carried out under the action of an acid-binding agent; the acid-binding agent is at least one selected from triethylamine, pyridine, N,N-diisopropylethylamine, sodium carbonate, and potassium carbonate.

4. The preparation method according to claim 3, characterized in that: The molar ratio of 2-aminobenzamide to methyl 5-bromopentanoate and the acid-binding agent is 1: (1-1.1): (2-3).

5. The preparation method according to claim 2, characterized in that: The ring-closing reaction is carried out in the presence of a catalyst, which is at least one of p-toluenesulfonic acid and benzenesulfonic acid.

6. The preparation method according to claim 2, characterized in that: The molar ratio of intermediate A to substituted benzaldehyde is 1:(1-1.1).

7. The preparation method according to claim 2, characterized in that: The hydrolysis reaction is carried out under alkaline conditions, provided by at least one of sodium carbonate, potassium carbonate, sodium hydroxide, and potassium hydroxide.

8. The preparation method according to claim 2, characterized in that: The amide condensation reaction is carried out in the presence of a condensing agent; the condensing agent is at least one selected from 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, carbonyl diimidazole, O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethylurea hexafluorophosphate, benzotriazol-N,N,N′,N′-tetramethylurea hexafluorophosphate, and 1-n-propylphosphoric anhydride.

9. The preparation method according to claim 2, characterized in that: The molar ratio of intermediate C to O-benzylhydroxylamine hydrochloride is 1:(1-1.1).

10. The preparation method according to claim 2, characterized in that: The hydrogenation reaction is carried out in the presence of a catalyst; the catalyst is at least one of palladium on carbon, platinum on carbon, and Raney nickel, and the hydrogen pressure is 30-50 Psi.

11. The use of the N-hydroxyamide compound containing dihydroquinazolinone as described in claim 1 in the preparation of histone deacetylase 6 inhibitors.

12. The use of the N-hydroxyamide compound containing dihydroquinazolinone as described in claim 1 in the preparation of medicaments for the prevention and / or treatment of diseases associated with abnormal expression of histone deacetylase 6 activity.

13. The application according to claim 12, characterized in that: The disease associated with abnormal expression of histone deacetylase 6 activity is cervical cancer.

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