HDACs inhibitor and preparation method and application thereof

Abstract

The present application belongs to the field of HDACs inhibitors, and relates to a HDACs inhibitor and a preparation method and application thereof. The structure of the inhibitor is shown in formula (1), wherein R1 is a different fatty or aromatic linking group, and R2 is a hydroxyl group or a phenyl group substituted with an adjacent amino group. The compound of the present application can effectively inhibit the activity of histone deacetylase (HDACs).

Description

Technical Field

[0001] This invention belongs to the field of HDACs inhibitors, and specifically relates to an HDACs inhibitor, its preparation method, and its application. Background Technology

[0002] The information disclosed in this background section is intended only to enhance understanding of the overall background of the invention and is not necessarily to be construed as an admission or in any way implying that such information constitutes prior art known to those skilled in the art.

[0003] Studies have revealed that HDACs induce various cellular processes through deacetylation of histone or non-histone substrates, thereby influencing the occurrence or progression of a variety of diseases, especially cancer. Tumors are closely associated with the overexpression of HDACs, and the development of drugs with HDAC inhibitory activity holds broad application prospects.

[0004] Currently, HDAC inhibitors are believed to consist of three main parts: the enzyme surface binding region (Cap group), the linker, and the zinc binding group (ZBG). The Cap group can interact with certain amino acid residues (usually aromatic groups) on the target enzyme surface. The linker is the bridge connecting the Cap and ZBG. On the one hand, it improves the selectivity of the enzyme by occupying the hydrophobic cavity of the enzyme; on the other hand, it affects the degree of binding between the zinc ion chelating group and zinc ions, thus affecting the inhibitor's activity on the enzyme. The ZBG is the key pharmacophore of histone deacetylase inhibitors. Common groups with strong chelating effects include hydroxamic acid and o-phenylenediamine.

[0005] Several histone deacetylase inhibitors are currently marketed both domestically and internationally. Of the four FDA-approved drugs, vorinostat (SAHA), panobinostat (LBH589), and belinostat (PXD101) all contain an isohydroxamic acid structure; romidepsin (FK228) is a cyclic peptide histone deacetylase inhibitor. Chidamide (CS055) has been approved for marketing in China for the treatment of relapsed or refractory peripheral T-cell lymphoma.

[0006] While HDAC inhibitors have achieved significant breakthroughs in the field of anti-tumor therapy, approved HDAC inhibitors suffer from problems such as significant clinical side effects and poor efficacy in treating solid tumors. Therefore, developing novel HDAC inhibitors with low toxicity and high efficacy is of great value for cancer treatment. Summary of the Invention

[0007] To address the aforementioned problems, this invention provides a novel HDACs inhibitor, its preparation method, and its application.

[0008] To achieve the above objectives, the present invention adopts the following technical solution:

[0009] In a first aspect, the present invention provides an HDACs inhibitor, said inhibitor being a compound represented by formula (1), a pharmaceutically acceptable salt, stereoisomer, isotope label, solvate, polycrystalline or prodrug thereof.

[0010]

[0011] In this context, R1 represents different aliphatic or aromatic linking groups, and R2 represents a phenyl group substituted with a hydroxyl or ortho-amino group.

[0012] In some embodiments, R1 is selected from the following groups:

[0013]

[0014] More specifically, the present invention provides compounds represented by formula (2) or formula (3), which are pharmaceutically acceptable salts, stereoisomers, isotope-labeled compounds, solvates, polycrystalline or prodrugs.

[0015]

[0016] Where R is

[0017]

[0018] In some embodiments, the inhibitor is selected from the following compounds:

[0019] .

[0020] Furthermore, the compound is one of J1, J4, J8, J16, or J17.

[0021] Furthermore, the compounds are J8 and J17.

[0022] In a second aspect, the present invention provides a method for preparing the above-mentioned HDACs inhibitor, the synthetic route of which is as follows:

[0023]

[0024] Wherein, 5a-b refers to 5a or 5b, 6a-b refers to 6a or 6b, 7a-j refers to 7a, 7b, 7c, 7d, 7e, 7f, 7g, 7h, 7i or 7j, and J1-10 refers to J1, J2, J3, J4, J5, J6, J7, J8, J9 or J10; the naming rules for other compounds in the synthetic route are the same as above, and will not be repeated here.

[0025]

[0026] In some embodiments, condition a involves adding LDA, Pd(dba)2, p(t-Bu)3, and toluene, and reacting overnight at 35-40°C.

[0027] In some embodiments, condition b is the addition of NaOH, MeOH / H2O (v / v) = 1:1, and reaction at 65-70°C for 3 hours.

[0028] In some implementations, condition c involves the addition of SOCl2, DIPEA, DCM, and DMF, followed by overnight reaction at room temperature.

[0029] In some embodiments, condition d is the addition of NH2OH, KOH, and MeOH, followed by a reaction at room temperature for 1 hour.

[0030] In some embodiments, condition e is the addition of KOH and MeOH, and the reaction at 75-80°C for 2-3 hours;

[0031] In some embodiments, the f condition is the addition of EDCI, DMAP, o-phenylenediamine, and THF, followed by a reaction at room temperature for 2-3 hours;

[0032] In some embodiments, the g condition is the addition of SnCl2 and ethanol, and the reaction at 60-70°C for 3.5-4 hours;

[0033] In some implementations, the h condition is the addition of SOCl2 and MeOH, and the reaction is carried out at room temperature for 5-6 hours.

[0034] The present invention also provides the application of the above-described HDACs inhibitors or the HDACs inhibitors prepared by the above-described methods in the preparation of antitumor drugs.

[0035] Beneficial effects of the present invention

[0036] This invention synthesized 30 novel HDAC inhibitors, and the structures of all compounds were subjected to [further testing / analysis]. 1 H NMR, 13 Confirmation by C10 NMR and ESI-MS. In vitro evaluation of the enzyme-inhibiting activity of HDACs showed that these compounds exhibited varying degrees of inhibition against the enzyme, with eight compounds showing IC50 values ​​of [missing information]. 50With values ​​below 100 nM, four compounds outperformed the marketed clinical drugs PXD101 and SAHA. Of the isohydroxamic acid compounds, except for eight with relatively poor activity, the remaining compounds showed good IC50 values. 50 The values ​​were all below 1000 nM. However, when selecting isohydroxamic acid compounds with better activity, the activity decreased sharply after replacing isohydroxamic acid ZBG with o-phenylenediamine. This may be because isohydroxamic acid has a better metal chelating ability than o-phenylenediamine, and therefore can chelate well with zinc ions in the HDAC pocket, thus enhancing the activity. J8, with a 2,5-substituted thiazole group as the linker and isohydroxamic acid as ZBG, exhibited the strongest inhibitory activity (IC50). 50 =0.76 nM). When the thiazole in the linker group changes from 2,5-substituted to 2,4-substituted (J7), the inhibitory activity decreases sharply (IC50). 50 =5733.5nM). Replacing S atoms with O atoms (J9) also leads to a decrease in activity (IC). 50 =10836nM). Replacing N atoms with C atoms (J17) exhibits the second highest inhibitory activity after J8, IC 50 The value is 1.7 nM. J18(IC) 50 The difference between J15 (3501 nM) and J17 lies in the substitution sites on thiophene, with 2,5-substitution being superior to 2,4-substitution. For compounds with benzene or pyridine as linkers, para-substitution is more active than meta-substitution. J15 (IC 50 =68.8nM) and J21(IC 50 Linkers with a molecular weight greater than 25000 nM exhibit vastly different isomers with varying activities, which may be related to the structure of adjacent hydrophobic protein channels. Adding a methylene group (J10) between the Cap and the benzene ring leads to a sharp decrease in activity (IC50). 50 =9791.5 nM). When the linker portion consists of fatty chains of varying lengths (J11-J14), compounds with four-carbon fatty chain substitution exhibit the best activity. Evaluation of antitumor cell proliferation activity also showed that the five compounds with strong enzyme inhibition activity all had higher antiproliferative activity than the marketed clinical drugs SAHA and PXD101. Compounds J8 and J17 showed superior antiproliferative activity compared to the lead compound JT86, while compound J8 showed an IC50 of 9791.5 nM against five types of solid tumors. 50 The values ​​are all less than 100 nM, therefore, this compound deserves further investigation to address the issue of poor efficacy of approved HDAC inhibitors in the treatment of solid tumors. Detailed Implementation

[0037] It should be noted that the following detailed descriptions are exemplary and intended to provide further illustration of the invention. Unless otherwise specified, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0038] The present invention will be further described in detail below with reference to specific embodiments. It should be noted that the specific embodiments are explanations of the present invention and not limitations thereof.

[0039] Example 1

[0040] Synthesis of target compounds J1-J21

[0041] 4-Naphthalene-tetrahydropyran-4-carboxylic acid methyl ester (3)

[0042] Compound 2 (2.30 g, 15.94 mmol) was dissolved in 100 mL of toluene in a double-necked flask under nitrogen protection. LDA (2.02 g, 18.83 mmol) was slowly added dropwise, and the mixture was reacted at room temperature for 10 min. Then, compound 1 (3.00 g, 14.49 mmol) and bis(di-benzylacetone)palladium (0.42 g, 0.72 mmol) were dissolved in 50 mL of toluene and slowly added to the reaction flask. Tri-tert-butylphosphine (0.29 g, 1.45 mmol) was then slowly added, and the mixture was reacted overnight at 35 °C. After the reaction was complete, the solvent was removed under reduced pressure, the reaction was quenched with saturated ammonium chloride solution, and the mixture was extracted three times with DCM. The organic phase was extracted twice with saturated sodium chloride solution. The mixture was dried over an appropriate amount of anhydrous magnesium sulfate, filtered, and the solvent was removed under reduced pressure. The product was purified by column chromatography to obtain a white solid product 3 (2.07 g, yield 52.9%).

[0043] 4-Naphthalene-tetrahydropyran-4-carboxylic acid (4)

[0044] Product 3 (2.07 g, 7.67 mmol) was added to 50 mL of methanol:water (v:v) = 1:1 in a round-bottom flask to form a suspension. NaOH (0.92 g, 23.01 mmol) was added, and the mixture was heated to reflux at 65 °C for 3 h. After the reaction was complete, methanol was removed under reduced pressure, the pH was adjusted to 1 with 2 M HCl, and the mixture was extracted three times with EA. The organic phase was extracted twice with saturated sodium chloride solution. The mixture was dried with an appropriate amount of anhydrous magnesium sulfate, filtered, and the solvent was removed under reduced pressure to give a white solid product 4 (1.92 g, 97.7%). 1H NMR (600MHz, DMSO-d6) δ12.67(s,1H),7.96-7.93(m,1H),7.90(ddd,J=11.8,6.5,3.4Hz,3H),7.58(dd,J=8.8,2.0Hz,1H),7.51(tt,J= 7.0,5.1Hz,2H),3.86(dt,J=11.8,3.8Hz,2H),3.52(td,J=11.3,2.1Hz,2H),2.49(s,2H),1.96(ddd,J=13.2,11.0,4.2Hz,2H).ESI-MS m / z: 255.1[MH] - .

[0045] (E)-4-aminocinnamate ethyl ester (6a)

[0046] Compound 5a (1.00 g, 4.52 mmol) and stannous chloride (4.28 g, 22.6 mmol) were added to 60 mL of ethanol and placed in a round-bottom flask. The mixture was heated to reflux at 60 °C for 3.5 h. After the reaction was complete, about half of the solvent was removed by vacuum distillation. The mixture was then poured into ice water, and the pH was adjusted to 7 with saturated NaHCO3 solution. The mixture was extracted three times with EA, and the organic phase was extracted twice with saturated sodium chloride solution. The mixture was dried over an appropriate amount of anhydrous magnesium sulfate, filtered, and the solvent was removed by vacuum distillation. Column chromatography purification yielded a yellow solid product 6a (0.69 g, yield 80.3%).

[0047] (E)-3-aminocinnamate ethyl ester (6b)

[0048] The synthesis method of intermediate 6b is the same as that of product 6a. It is a yellow solid with a yield of 93.9%.

[0049] 8-Aminooctanoic acid methyl ester (10a)

[0050] Thionyl chloride (0.93 g, 7.85 mmol) was added dropwise to 10 mL of methanol and placed in an ice bath. After reacting for 10 min, intermediate 9a (0.50 g, 3.14 mmol) was added and reacted at room temperature for 5 h. The solvent was removed by vacuum evaporation to obtain a white solid crude product 10a.

[0051] Methyl 7-aminoheptanoate (10b)

[0052] The synthesis method of intermediate 10b is the same as that of product 10a. It is a white solid.

[0053] Methyl 6-aminohexanoate (10c)

[0054] The synthesis method of intermediate 10c is the same as that of product 10a. It is a white solid.

[0055] Methyl 5-aminovalerate (10d)

[0056] The synthesis method of intermediate 10d is the same as that of product 10a. It is a white solid.

[0057] (E)-N-(4-(3-(hydroxyamino)-3-oxopropyl-1-en-1-yl)phenyl)-4-(naphth-2-yl)tetrahydro-2H-pyran-4-carboxamide (J1)

[0058] Product 4 (0.20 g, 0.78 mmol) was placed in a two-necked flask, and 5 mL of thionyl chloride was added. Under nitrogen protection, the mixture was heated to reflux at 80 °C for 4 h. The thionyl chloride was removed by vacuum distillation. The residue was washed twice with redistilled DCM, and the solvent was removed by vacuum distillation. Redistilled DCM was added to prepare solution a. Separately, product 6a (0.18 g, 0.94 mmol) was dissolved in redistilled DCM and placed in a two-necked flask to prepare solution b. Solution a was added dropwise to solution b, followed by DIPEA (0.30 g, 2.34 mmol) and 0.5 mL of DMF. The mixture was reacted overnight at room temperature under nitrogen protection. After the reaction was completed, the product was extracted twice each with 2 M HCl, saturated NaHCO3 solution, and saturated sodium chloride solution. The product was dried over an appropriate amount of anhydrous magnesium sulfate, filtered, and the solvent was removed by vacuum distillation to obtain crude product 7a.

[0059] Preparation of NH₂OK solution: Dissolve 14.18 g KOH in 35 mL MeOH under ice bath to prepare solution a. Dissolve 11.68 g hydroxylamine hydrochloride in 60 mL MeOH to prepare solution b. Add solution a dropwise to solution b, stir for 1 h, and filter to remove the precipitate to obtain the NH₂OK solution. Dissolve the crude product 7a in 12 mL NH₂OK solution and react at room temperature for 30 min. After the reaction is complete, remove the solvent under reduced pressure, add an appropriate amount of water, adjust the pH to 1 with 2M HCl, extract three times with EA, extract the organic phase twice with saturated sodium chloride solution, add an appropriate amount of anhydrous magnesium sulfate to dry, filter, remove the solvent under reduced pressure, and purify by HPLC to obtain the yellow solid product J1 (0.042 g, 12.8%). Mp: 109-111℃. 1H NMR (600MHz, DMSO-d6) δ10.62(s,1H),9.42(s,1H),8.99(s,1H),7.99-7.93(m,2H),7.93-7. 85(m,2H),7.62(d,J=8.4Hz,2H),7.57(dd,J=8.7,2.0Hz,1H),7.51(dqd,J=8.3,6.9,1.5Hz, 2H),7.45(d,J=8.4Hz,2H),7.35(d,J=15.8Hz,1H),6.34(d,J=15.8Hz,1H),3.85(dt,J=11.7 ,3.8Hz,2H),3.66-3.55(m,2H),2.69(d,J=13.6Hz,2H),2.11(ddd,J=14.2,10.6,4.0Hz,2H). 13 C NMR(151MHz,DMSO-d6)δ173.17,163.76,141.64,140.83,138.63,133.76,132.72,130.97,128.95,12 8.87,128.60,128.13,127.06,126.87,125.34,125.01,121.45,118.53,65.43,50.17,35.04.ESI-MS m / z: 415.4[MH] - .

[0060] (E)-N-(3-(3-(hydroxyamino)-3-oxopropyl-1-en-1-yl)phenyl)-4-(naphth-2-yl)tetrahydro-2H-pyran-4-carboxamide (J2)

[0061] The synthesis method for J2 is the same as that for J1. It is a white solid with a yield of 3.1%. Mp: 134-136℃. 1H NMR(600MHz,DMSO-d6)δ10.72(s,1H),9.38(s,1H),9.04(s,1H),8.00-7.93(m,2H),7.91(d,J =8.7Hz,1H),7.89-7.83(m,2H),7.58(dd,J=8.7,2.0Hz,1H),7.55-7.47(m,3H),7.36(d,J=15. 8Hz,1H),7.29(t,J=7.9Hz,1H),7.20(d,J=7.6Hz,1H),6.40(d,J=15.8Hz,1H),3.86(dt,J=11. 8,3.9Hz,2H),3.68-3.55(m,2H),2.70(d,J=13.7Hz,2H),2.11(ddd,J=14.2,10.6,4.0Hz,2H). 13 C NMR(151MHz,DMSO-d6)δ173.20,163.33,141.72,140.22,138.90,135.93,133.76,132.72,129.85,128.95,12 8.88,128.14,127.07,126.86,125.32,125.02,124.53,122.57,120.12,119.28,65.44,50.09,35.06.ESI-MS m / z:415.4[MH] - .

[0062] N-(3-(hydroxycarbamoyl)phenyl)-4-(naphth-2-yl)tetrahydro-2H-pyran-4-carboxamide (J3)

[0063] The synthesis method for J3 is the same as that for J1. It is a brownish-yellow solid with a yield of 47.9%. Mp: 122-124℃. 1 H NMR(600MHz,DMSO-d6)δ11.13(s,1H),9.45(s,1H),8.99(s,1H),8.01-7.85(m ,5H),7.77(d,J=7.4Hz,1H),7.58(dd,J=8.7,1.9Hz,1H),7.56-7.45(m,2H),7. 38(d,J=7.7Hz,1H),7.31(t,J=7.9Hz,1H),3.85(dt,J=11.7,3.9Hz,2H),3.62( t,J=11.0Hz,2H),2.70(d,J=13.6Hz,2H),2.10(ddd,J=14.3,10.7,4.1Hz,2H). 13C NMR(151MHz,DMSO-d6)δ172.29,163.97,140.82,139.01,133.16,132.87,131.83,128.34,128.04,12 7.99,127.25,126.18,125.98,124.47,124.15,123.03,121.59,119.63,64.56,49.16,34.15.ESI-MS m / z: 389.4[MH] - .

[0064] N-Hydroxy-6-(4-(naphth-2-yl)tetrahydro-2H-pyran-4-carboxylamine)nicotinamide (J4)

[0065] The synthesis method for J4 is the same as that for J1. It is a brownish-yellow solid with a yield of 25.8%. Mp: 115-116℃. 1 H NMR(600MHz,DMSO-d6)δ11.22(s,1H),9.98(s,1H),9.14(d,J=77.4Hz,1H),8.64-8.53(m ,1H),8.15-8.06(m,2H),8.01(d,J=1.9Hz,1H),7.95(d,J=8.0Hz,1H),7.92-7.84(m,2H) ,7.57(dd,J=8.7,2.0Hz,1H),7.52(dddd,J=16.5,8.1,6.8,1.4Hz,2H),3.82(dt,J=11.9 ,4.1Hz,2H),3.67-3.58(m,2H),2.80-2.69(m,2H),2.14(ddd,J=14.0,10.1,3.9Hz,2H). 13 CNMR(151MHz,DMSO-d6)δ173.28,162.10,153.64,146.54,139.87,136.75,132.85,131.90,128.1 0,128.04,127.27,126.23,126.10,125.11,124.21,124.13,113.49,64.43,49.41,33.77.ESI-MS m / z: 390.4[MH] - .

[0066] N-Hydroxy-2-(4-(Naphth-2-yl)tetrahydro-2H-pyran-4-carboxylamine)isonicotinamide (J5)

[0067] The synthesis method for J5 is the same as that for J1. It is a yellow solid with a yield of 13.1%. Mp: 95-96℃. 1H NMR (600MHz, DMSO-d6) δ11.39(s,1H),9.90(s,1H),9.32(s,1H),8.35(d,J=1.5Hz,1H),8.31(d,J= 5.1Hz,1H),8.01(d,J=2.0Hz,1H),7.97-7.93(m,1H),7.93-7.86(m,2H),7.59(dd,J=8.7,2.0Hz,1 H),7.52(dddd,J=16.5,8.2,6.9,1.5Hz,2H),7.33(dd,J=5.1,1.5Hz,1H),3.83(dt,J=11.8,4.0Hz ,2H),3.69-3.60(m,2H),2.76(ddd,J=12.1,4.7,2.5Hz,2H),2.14(ddd,J=14.0,10.2,3.9Hz,2H). 13 C NMR(151MHz,DMSO-d6)δ173.92,162.98,153.30,148.97,142.97,140.93,133.74,132.77,128.9 4,128.90,128.14,127.08,126.94,125.92,125.12,117.64,113.25,65.33,50.18,34.71.ESI-MS m / z: 390.4[MH] - .

[0068] N-hydroxy-6-(4-(naphth-2-yl)tetrahydro-2H-pyran-4-carboxamine)pyrazine-3-carboxamide (J6)

[0069] The synthesis method for J6 is the same as that for J1. It is a yellow solid with a yield of 6.5%. Mp: 202-203℃. 1 H NMR (600MHz, DMSO-d6) δ11.61(s,1H),10.77(s,1H),9.19(s,1H),8.41(d,J=9.3Hz ,1H),8.10(d,J=9.3Hz,1H),8.04-8.02(m,1H),7.97(d,J=8.0Hz,1H),7.94-7.87( m,2H),7.58(dd,J=8.7,2.0Hz,1H),7.56-7.49(m,2H),3.85(dt,J=12.0,3.9Hz,2H ),3.69-3.61(m,2H),2.78(d,J=13.8Hz,2H),2.16(ddd,J=14.1,10.3,4.0Hz,2H). 13C NMR(151MHz,DMSO-d6)δ174.93,160.78,157.53,151.23,140.53,133.73,132.82,129.02,12 8.92,128.16,127.95,127.13,127.01,126.05,125.05,121.00,65.29,50.50,34.71.ESI-MS m / z: 391.4[MH] - .

[0070] N-Hydroxy-2-(4-(naphthyl-2-yl)tetrahydro-2H-pyran-4-carboxamide)thiazole-4-carboxamide (J7)

[0071] J7 was synthesized using the same method as J1. It is a white solid with a yield of 39.9%. Mp: 132-134℃. 1 H NMR(600MHz,DMSO-d6)δ12.03(s,1H),10.62(s,1H),9.01(s,1H),8.00-7.93(m,2H),7.93-7.85(m,2H),7.69(s,1H),7.52(dtd, J=9.8,6.8,5.5Hz,3H),3.84(dt,J=11.9,4.0Hz,2H),3.61-3.52(m,2H),2.81-2.71(m,2H),2.15(ddd,J=14.2,10.2,3.9Hz,2H). 13 C NMR(151MHz,DMSO-d6)δ174.05,160.16,159.16,143.88,140.30,133.67,132.80,129.0 3,128.92,128.14,127.16,127.06,126.01,124.95,117.52,65.26,49.63,34.42.ESI-MS m / z: 396.4[MH] - .

[0072] N-Hydroxy-2-(4-(naphthyl-2-yl)tetrahydro-2H-pyran-4-carboxamine)thiazole-5-carboxamide (J8)

[0073] J8 was synthesized using the same method as J1. It is a white solid with a yield of 30.0%. Mp: 219-220℃. 1H NMR (600MHz, DMSO-d6) δ12.25(s,1H),11.08(s,1H),9.09(s,1H),7.96(d,J=1.9Hz,1H),7.95(d,J=7.9Hz,1H),7.89(dd,J=15.3,8.3Hz,2H), 7.85(s,1H),7.57-7.45(m,3H),3.83(dt,J=11.9,4.0Hz,1H),3.60-3.51(m,2H),2.75(d,J=13.7Hz,2H),2.15(ddd,J=14.1,10.4,4.0Hz,2H). 13 C NMR(151MHz,DMSO-d6)δ173.66,161.16,159.59,139.81,139.01,133.30,132.45,128.72 ,128.58,127.82,126.85,126.76,125.70,124.58,64.90,49.33,34.03,tautomerisation broadened signals 1C not seen.ESI-MS m / z: 396.4[MH] - .

[0074] N-Hydroxy-2-(4-(Naphth-2-yl)tetrahydro-2H-pyran-4-carboxylamine)oxazole-5-carboxamide (J9)

[0075] J9 was synthesized using the same method as J1. It is a brownish-yellow solid with a yield of 17.5%. Mp: 109-110℃. 1 H NMR(600MHz,DMSO-d6)δ11.08(s,2H),9.17(s,1H),7.98-7.88(m,4H),7.57-7.47(m,4H),3.83(dt, J=11.9,3.9Hz,2H),3.64-3.57(m,2H),2.64(d,J=13.7Hz,2H),2.09(ddd,J=14.4,10.5,3.9Hz,2H). 13 C NMR(151MHz,DMSO-d6)δ140.73,140.47,133.71,132.82,129.00,128.90,128.77 ,128.16,127.13,127.02,125.86,125.00,65.25,50.22,34.70,tautomerisation broadened signals 3C not seen.ESI-MS m / z: 380.3[MH]- .

[0076] N-(4-(hydroxycarbamoyl)benzyl)-4-(naphth-2-yl)tetrahydro-2H-pyran-4-carboxamide (J10)

[0077] The synthesis method for J10 is the same as that for J1. It is a white solid with a yield of 27.5%. Mp: 192-193℃. 1 H NMR (600MHz, DMSO-d6) δ11.08(s,1H),8.98(s,1H),8.28(t,J=6.0Hz,1H),7.93-7.84(m,4H),7.57-7.46(m,5H),7.07(d,J=8.0Hz,2 H),4.27(d,J=5.9Hz,2H),3.79(dt,J=11.7,3.9Hz,2H),3.57-3.51(m,2H),2.61-2.55(m,2H),2.01(ddd,J=14.0,10.4,4.0Hz,2H). 13 C NMR(151MHz,DMSO-d6)δ173.16,164.01,143.11,141.51,132.89,131.81,131.05,127.99,127.8 4,127.30,126.67,126.64,126.16,125.94,124.35,124.32,64.64,48.05,42.16,34.10.ESI-MS m / z: 403.4[MH] - .

[0078] N-(8-(hydroxyamino)-8-oxooctyl)-4-(naphth-2-yl)tetrahydro-2H-pyran-4-carboxamide (J11)

[0079] J11 was synthesized using the same method as J1. It is a white solid with a yield of 19.4%. Mp: 116-118℃. 1H NMR(600MHz,DMSO-d6)δ10.30(s,1H),8.64(s,1H),7.90(dd,J=7.8,1.5Hz,1H),7.89-7.80(m,3H ),7.62(t,J=5.8Hz,1H),7.54-7.44(m,3H),3.79(dt,J=11.6,3.8Hz,2H),3.52(td,J=11.1,2.0Hz ,2H),3.18(d,J=5.1Hz,2H),3.01(q,J=6.5Hz,2H),2.59-2.52(m,2H),1.95(ddd,J=14.2,10.7,4 .0Hz,2H),1.88(t,J=7.4Hz,2H),1.38(p,J=7.4Hz,2H),1.30(p,J=7.1Hz,2H),1.15-0.98(m,4H). 13 C NMR(151MHz,DMSO-d6)δ173.51,169.98,142.75,133.71,132.59,128.75,128.57,128.06,126.87,12 6.61,125.08,124.90,65.50,48.86,39.69,35.02,33.05,29.70,29.30,29.19,26.94,25.81.ESI-MS m / z: 411.5[MH] - .

[0080] N-(7-(hydroxyamino)-7-oxohepyl)-4-(naphth-2-yl)tetrahydro-2H-pyran-4-carboxamide (J12)

[0081] J12 was synthesized using the same method as J1. It is a white solid with a yield of 77.7%. Mp: 141-142℃. 1H NMR (600MHz, DMSO-d6) δ10.29(s,1H),8.63(s,1H),7.90(dd,J=7.8,1.6Hz,1H),7.86(dd,J=8.3,2.2Hz,2H) ,7.84(d,J=1.9Hz,1H),7.62(t,J=5.7Hz,1H),7.53-7.46(m,3H),3.79(dt,J=11.7,3.8Hz,2H),3.52(td,J= 11.2, 2.0Hz, 2H), 3.01 (q, J=6.6Hz, 2H), 2.58-2.51 (m, 2H), 1.94 (ddd, J=14.2, 10.7, 4.1Hz, 2H), 1.85 (t, J= 7.4Hz,2H),1.35(q,J=7.4Hz,2H),1.29(q,J=7.1Hz,2H),1.11(q,J=7.4,7.0Hz,2H),1.07(q,J=7.5Hz,2H). 13 C NMR(151MHz,DMSO-d6)δ172.61,169.03,141.86,132.82,131.69,127.87,127.68,127.17,126.00 ,125.74,124.21,124.01,64.61,47.97,38.80,34.16,32.15,28.74,28.19,25.91,24.95.ESI-MS m / z: 397.5[MH] - .

[0082] N-(6-(hydroxyamino)-6-oxohexyl)-4-(naphth-2-yl)tetrahydro-2H-pyran-4-carboxamide (J13)

[0083] J13 was synthesized using the same method as J1. It is a white solid with a yield of 42.4%. Mp: 76-77℃. 1H NMR(600MHz,DMSO-d6)δ10.29(s,1H),8.64(s,1H),7.91(dd,J=7.8,1.6Hz,1H),7.89-7.81 (m,3H),7.63(t,J=5.7Hz,1H),7.54-7.45(m,3H),3.79(dt,J=11.6,3.8Hz,2H),3.52(td,J= 11.2,2.0Hz,2H),3.00(q,J=6.6Hz,2H),2.58-2.51(m,2H),1.95(ddd,J=14.2,10.7,4.0Hz, 2H),1.84(t,J=7.5Hz,2H),1.39(p,J=7.5Hz,2H),1.31(t,J=7.5Hz,2H),1.15-1.00(m,2H). 13 C NMR(151MHz,DMSO-d6)δ172.63,168.97,141.85,132.82,131.69,127.88,127.69,127.18,126 .00,125.74,124.24,124.02,64.61,47.97,38.77,34.17,32.10,28.60,25.85,24.69.ESI-MS m / z: 383.4[MH] - .

[0084] N-(5-(hydroxyamino)-5-oxopentyl)-4-(naphth-2-yl)tetrahydro-2H-pyran-4-carboxamide (J14)

[0085] J14 was synthesized using the same method as J1. It is a yellow solid with a yield of 56.1%. Mp: 69-70℃. 1 H NMR(600MHz,DMSO-d6)δ10.28(s,1H),8.63(s,1H),7.91(dd,J=7.6,1.8Hz,1H),7.89- 7.82(m,3H),7.67(t,J=5.7Hz,1H),7.54-7.45(m,3H),3.78(dt,J=11.6,3.7Hz,2H),3 .52(td,J=11.3,2.0Hz,2H),3.00(q,J=6.5Hz,2H),2.57-2.51(m,2H),1.94(ddd,J=14 .2,10.8,4.2Hz,2H),1.88(t,J=7.1Hz,2H),1.38(p,J=7.3Hz,2H),1.35-1.27(m,2H). 13C NMR(151MHz,DMSO-d6)δ173.52,169.81,142.75,133.71,132.56,128.78,128.60,128.07, 126.90,126.63,125.12,124.91,65.49,48.86,39.56,35.10,32.65,29.41,23.33.ESI-MS m / z: 369.4[MH] - .

[0086] N-((1r,4r)-4-(hydroxycarbamate)cyclohexyl)-4-(naphth-2-yl)tetrahydro-2H-pyran-4-carboxamide (J15)

[0087] J15 was synthesized using the same method as J1. It is a white solid with a yield of 67.5%. Mp: 216-217℃. 1 H NMR (600MHz, DMSO-d6) δ10.30(s,1H),8.61(s,1H),7.91(dd,J=7.7,1.6Hz,1H),7.86(dd,J=8.2,3.4Hz ,2H),7.83(d,J=1.9Hz,1H),7.55-7.45(m,3H),7.37(d,J=8.1Hz,1H),3.80(dt,J=11.7,3.5Hz,2H),3. 57(ddd,J=11.7,7.9,3.9Hz,1H),3.51(td,J=11.5,2.0Hz,2H),2.57-2.51(m,2H),1.93(ddd,J=14.2,1 1.0,4.1Hz,2H),1.88-1.79(m,1H),1.66-1.56(m,4H),1.45-1.33(m,2H),1.13(qd,J=12.8,3.7Hz,2H). 13 C NMR(151MHz,DMSO-d6)δ171.84,171.77,141.99,132.81,131.66,127.88,127.65,127.17, 125.98,125.72,124.19,123.95,64.62,48.02,47.55,40.38,34.26,31.06,28.07.ESI-MS m / z: 395.4[MH] - .

[0088] N-Hydroxy-5-(4-(naphth-2-yl)tetrahydro-2H-pyran-4-carboxamine)-1H-pyrazole-3-carboxamide (J16)

[0089] The synthesis method for J16 is the same as that for J1. It is a brownish-yellow solid with a yield of 5.5%. Mp: 214-215℃. 1 H NMR (600MHz, DMSO-d6) δ13.09(s,1H),11.17(s,1H),10.05(s,1H),9.11(s,1H),7.99-7.91(m,2H),7.88(t,J=8.4Hz,2H),7.55(dd,J=8 .7,1.9Hz,1H),7.54-7.46(m,2H),7.00(s,1H),3.82(dt,J=11.7,3.8Hz,2H),3.63-3.54(m,2H),2.78-2.63(m,2H),2.13-2.00(m,2H). 13 CNMR(151MHz,DMSO-d6)δ171.58,140.73,132.87,131.87,128.03,127.99,127.78 ,127.32,126.25,126.05,124.69,124.26,64.62,48.73,33.98,tautomerisation broadenedsignals 3C not seen.ESI-MS m / z: 379.4[MH] - .

[0090] N-(5-(hydroxycarbamoyl)thiophen-2-yl)-4-(naphthyl-2-yl)tetrahydro-2H-pyran-4-carboxamide (J17)

[0091] The synthesis method for J17 is the same as that for J1. It is a brownish-yellow solid with a yield of 2.0%. Mp: 179-181℃. 1 H NMR(600MHz,DMSO-d6)δ10.89(s,2H),8.93(s,1H),8.00-7.93(m,2H),7.93-7.85(m,2H),7.57-7.47(m,3H),7.33(s,1H),6.67 (d,J=4.2Hz,1H),3.85(dt,J=11.9,3.8Hz,2H),3.58-3.49(m,2H),2.65(d,J=13.6Hz,2H),2.13(ddd,J=14.3,10.6,4.0Hz,2H). 13C NMR(151MHz,DMSO-d6)δ171.87,141.10,133.72,132.77,129.11,128.88,128.15 ,127.16,127.00,125.35,124.89,112.71,65.34,49.48,34.84,tautomerisation broadened signals 4Cnot seen.ESI-MS m / z: 395.4[MH] - .

[0092] N-(5-(hydroxycarbamoyl)thiophen-3-yl)-4-(naphthyl-2-yl)tetrahydro-2H-pyran-4-carboxamide (J18)

[0093] J18 was synthesized using the same method as J1. It is a white solid with a yield of 58.4%. Mp: 121-123℃. 1 H NMR (600MHz, DMSO-d6) δ11.14(s,1H),9.81(s,1H),9.03(s,1H),7.99-7.92(m,2H),7.92-7.84(m,2H),7.66(d,J=15.3Hz,2H),7 .57-7.46(m,3H),3.84(dt,J=11.7,3.9Hz,2H),3.63-3.53(m,2H),2.65(d,J=13.6Hz,2H),2.08(ddd,J=14.1,10.5,4.0Hz,2H). 13 C NMR(151MHz,DMSO-d6)δ171.62,159.56,140.90,136.66,134.84,132.91,131.89,128.16,12 8.06,127.34,126.30,126.09,124.41,124.19,122.42,114.24,64.62,48.86,34.16.ESI-MS m / z: 395.4[MH] - .

[0094] N-Hydroxy-5-(4-(naphth-2-yl)tetrahydro-2H-pyran-4-carboxylamine)pyridineamide (J19)

[0095] J19 was synthesized using the same method as J1. It is a white solid with a yield of 7.6%. Mp: 198-199℃. 1H NMR (600MHz, DMSO-d6) δ11.25(s,1H),9.73(s,1H),9.03(d,J=67.7Hz,1H),8.78(d,J=2.4Hz,1H),8.17(dd,J=8.6,2.5Hz,1H),8.00-7.94(m,2H),7 .94-7.86(m,3H),7.59-7.45(m,3H),3.85(dt,J=11.8,3.8Hz,2H),3.66- 3.58(m,2H),2.68(d,J=13.7Hz,2H),2.14(ddd,J=14.4,10.6,4.1Hz,2H). 13 C NMR(151MHz,DMSO-d6)δ173.94,161.95,145.73,141.24,141.10,138.55,133.76,132.77,129.0 9,128.90,128.68,128.15,127.12,126.96,125.44,124.95,122.80,65.37,50.30,34.99.ESI-MS m / z: 390.4[MH] - .

[0096] N-Hydroxy-5-(4-(naphth-2-yl)tetrahydro-2H-pyran-4-carboxylamine)nicotinamide (J20)

[0097] The synthesis method for J20 is the same as that for J1. It is a brownish-yellow solid with a yield of 22.2%. Mp: 92-93℃. 1 H NMR (600MHz, DMSO-d6) δ11.34(s,1H),9.65(s,1H),9.17(s,1H),8.88(d,J=2.4Hz,1H),8. 56(d,J=1.9Hz,1H),8.33(t,J=2.2Hz,1H),7.97(dd,J=9.9,1.9Hz,2H),7.92(d,J=8.7Hz,1 H),7.90-7.87(m,1H),7.57(dd,J=8.7,2.0Hz,1H),7.55-7.49(m,2H),3.86(dt,J=11.9,3 .9Hz,2H),3.67-3.59(m,2H),2.69(d,J=13.7Hz,2H),2.13(ddd,J=14.1,10.5,4.0Hz,2H). 13C NMR(151MHz,DMSO-d6)δ172.98,162.32,143.87,142.32,140.50,135.52,132.92,131.92,128.3 8,128.24,128.08,127.33,126.30,126.21,126.13,124.57,124.13,64.55,49.29,34.15.ESI-MS m / z: 390.4[MH] - .

[0098] N-((1,4s)-4-(hydroxycarbamate)cyclohexyl)-4-(naphth-2-yl)tetrahydro-2H-pyran-4-carboxamide (J21)

[0099] The synthesis method for J21 is the same as that for J1. It is a brownish-yellow solid with a yield of 52.1%. Mp: 96-98℃. 1 H NMR(600MHz,DMSO-d6)δ10.30(s,1H),8.54(s,1H),7.93-7.88(m,1H),7.88-7.82(m,3H),7.55-7.51(m ,1H),7.49(td,J=7.5,1.5Hz,2H),7.31(d,J=7.1Hz,1H),3.80(dt,J=11.8,3.6Hz,2H),3.70(tq,J=7.5 ,3.8Hz,1H),3.51(td,J=11.3,1.9Hz,2H),2.63-2.56(m,2H),2.09(h,J=4.1,3.3Hz,1H),1.93(ddd,J= 14.6,11.2,4.1Hz,2H),1.76-1.66(m,2H),1.58(dtd,J=13.1,8.5,7.7,3.4Hz,2H),1.44-1.30(m,4H). 13 C NMR(125MHz,Chloroform-d)δ173.72,172.68,141.52,134.14,133.59,127.70,127.32,126. 99,126.74,126.31,126.26,125.91,66.02,49.63,48.64,40.21,33.36,30.47,25.94.ESI-MS m / z: 395.4[MH] - .

[0100] Synthesis of target compounds J22-J30

[0101] (E)-N-(4-(3-(2-aminophenyl)amino)-3-oxopropyl-1-en-1-yl)phenyl)-4-(naphth-2-yl)tetrahydro-2H-pyran-4-carboxamide (J22)

[0102] The crude product 7a was dissolved in 5 mL of methanol, and potassium hydroxide (0.13 g, 2.34 mmol) was added. The mixture was heated under reflux at 75 °C for 3 h. After the reaction was completed, methanol was removed by vacuum distillation, 10 mL of water was added, and the pH was adjusted to 1 with 2 M HCl. The mixture was extracted three times with EA, and the organic phase was extracted twice with saturated sodium chloride solution. The mixture was dried with an appropriate amount of anhydrous magnesium sulfate, filtered, and the solvent was removed by vacuum distillation to obtain the crude product 8a.

[0103] The crude product 8a was dissolved in 8 mL of THF, and EDCI (0.18 g, 0.94 mmol), DMAP (0.08 g, 0.62 mmol), and o-phenylenediamine (0.10 g, 0.94 mmol) were added sequentially. The reaction was carried out at room temperature for 2 h. After the reaction was completed, the product was extracted three times with water, and the organic phase was extracted twice with saturated sodium chloride solution. The product was dried over an appropriate amount of anhydrous magnesium sulfate, filtered, and the solvent was removed under reduced pressure. The product was purified by HPLC to obtain a yellow solid J22 (0.008 g, 4.7%). Mp: 209-211℃. 1 H NMR(600MHz,DMSO-d6)δ9.47(s,1H),9.35(s,1H),8.00-7.94(m,2H),7.94-7.86(m,2H),7.66(d,J=8.4Hz,2H),7 .57(dd,J=8.7,1.9Hz,1H),7.52(dtd,J=12.9,6.3,3.5Hz,4H),7.45(d,J=15.7Hz,1H),7.33(d,J=7.9Hz,1H),6. 93-6.86(m,1H),6.80(d,J=15.7Hz,1H),6.74(dd,J=8.0,1.4Hz,1H),6.56(td,J=7.5,1.4Hz,1H),4.94(s,2H),3 .86(dt,J=11.9,3.7Hz,2H),3.61(t,J=10.7Hz,2H),2.70(d,J=13.5Hz,2H),2.11(ddd,J=14.5,10.7,4.0Hz,2H). 13C NMR(151MHz,DMSO-d6)δ172.35,163.61,141.50,140.79,140.17,139.05,132.90,131.88,130.12,128.14,128.05,127.98,12 7.31,126.25,126.06,125.63,124.59,124.50,124.18,123.59,120.93,120.62,116.22,115.96,64.60,49.32,34.19.ESI-MS m / z: 490.5[MH] - .

[0104] N-(2-Aminophenyl)-6-(4-(naphth-2-yl)tetrahydro-2H-pyran-4-carboxylamine)nicotinamide (J23)

[0105] The synthesis method for J23 is the same as that for J22. It is a brownish-yellow solid with a yield of 5.2%. Mp: 122-124℃. 1 H NMR (600MHz, DMSO-d6) δ10.13(s,1H),9.73(s,1H),8.83(s,1H),8.33(d,J=8.8Hz,1H),8.17(d,J=8.9Hz ,1H),8.03(s,1H),7.96(d,J=8.0Hz,1H),7.90(dd,J=16.7,8.3Hz,2H),7.62-7.57(m,1H),7.53(dt,J=17 .2,7.2Hz,2H),7.12(d,J=7.8Hz,1H),6.96(t,J=7.7Hz,1H),6.76(d,J=8.0Hz,1H),6.57(t,J=7.6Hz,1H) ,4.93(s,2H),3.83(d,J=10.4Hz,2H),3.64(t,J=11.0Hz,2H),2.78(d,J=13.7Hz,2H),2.19-2.09(m,2H). 13 C NMR(151MHz,DMSO-d6)δ173.65,163.58,154.10,147.92,143.58,140.20,138.01,133.16,132.21,128.41,128.37,127.6 1,127.14,126.94,126.57,126.43,126.16,125.45,124.56,123.02,116.37,116.23,113.52,64.77,49.75,34.09.ESI-MS m / z: 465.5[MH] - .

[0106] N-(2-aminophenyl)-2-(4-(naphthyl-2-yl)tetrahydro-2H-pyran-4-carboxamide)thiazole-5-carboxamide (J24)

[0107] J24 was synthesized using the same method as J22. It is a white solid with a yield of 7.7%. Mp: 196-197℃. 1 H NMR (600MHz, DMSO-d6) δ12.17(s,1H),9.59(s,1H),8.21(s,1H),7.97(dd,J=7.6,4.9H z,2H),7.94-7.86(m,2H),7.59-7.47(m,3H),7.11(dd,J=7.8,1.5Hz,1H),6.99-6.93(m ,1H),6.76(dd,J=8.0,1.4Hz,1H),6.57(td,J=7.5,1.4Hz,1H),4.91(s,2H),3.85(dt, J=12.0,3.9Hz,2H),3.62-3.52(m,2H),2.76(d,J=13.6Hz,2H),2.16(t,J=11.3Hz,2H). 13 C NMR(151MHz,DMSO-d6)δ159.58,143.19,132.82,131.96,128.21,128.09,127.33,126.76,126.64,12 6.36,126.25,125.18,124.12,122.47,116.16,115.99,64.44,48.89,40.06,33.60,tautomerization broadened signals 4C not seen.ESI-MS m / z: 471.5[MH] - .

[0108] N-(8-(2-aminophenyl)amino)-8-oxooctyl)-4-(naphth-2-yl)tetrahydro-2H-pyran-4-carboxamide (J25)

[0109] J25 was synthesized using the same method as J22. It is a white solid with a yield of 15.4%. Mp: 135-136℃. 1H NMR(600MHz, DMSO-d6)δ9.07(d,J=4.3Hz,1H),7.90(d,J=7.4Hz,1H),7.89-7.82(m,3H),7.69-7.60(m,1H),7.56-7 .44(m,3H),7.20-7.10(m,1H),6.94-6.85(m,1H),6.70(d,J=7.7Hz,1H),6.53(q,J=7.5,6.2Hz,1H),4.80(d,J=4.2H z,2H),3.87-3.74(m,2H),3.51(dd,J=12.9,9.2Hz,2H),3.01(q,J=6.2Hz,2H),2.53(s,2H),2.24(q,J=7.3,6.5Hz, 2H),2.01-1.87(m,2H),1.56-1.42(m,2H),1.30(p,J=6.5Hz,2H),1.15(d,J=6.0Hz,4H),1.06(q,J=7.3,6.9Hz,2H). 13 C NMR(151MHz,DMSO-d6)δ172.48,170.99,141.80,141.69,132.72,131.59,127.79,127.59,127.10,125.92,125.66,125.51,12 5.08,124.11,123.89,123.46,116.03,115.75,64.54,47.85,38.69,35.58,34.03,28.75,28.43,28.32,25.99,25.04.ESI-MS m / z: 486.6[MH] - .

[0110] N-(7-(2-aminophenyl)amino)-7-oxohepyl)-4-(naphth-2-yl)tetrahydro-2H-pyran-4-carboxamide (J26)

[0111] J26 was synthesized using the same method as J22. It is a white solid with a yield of 53.2%. Mp: 10²-10⁴℃. 1H NMR(600MHz, DMSO-d6)δ9.06(s,1H),7.91(d,J=7.9Hz,1H),7.88-7.81(m,3H),7.65(t,J=5.8Hz,1H),7.54-7.43(m,3H),7.13 (dd,J=7.9,1.5Hz,1H),6.88(td,J=7.6,1.5Hz,1H),6.71(dd,J=8.0,1.4Hz,1H),6.53(td,J=7.6,1.4Hz,1H),4.80(s,2H),3.7 9(dt,J=11.6,3.8Hz,2H),3.52(t,J=11.0Hz,2H),3.02(q,J=6.6Hz,2H),2.55(d,J=13.4Hz,2H),2.22(t,J=7.5Hz,2H),1.94( ddd,J=14.2,10.8,4.0Hz,2H),1.44(p,J=7.5Hz,2H),1.32(p,J=7.1Hz,2H),1.20(p,J=7.4Hz,2H),1.11(q,J=7.7,7.3Hz,2H). 13 C NMR(151MHz,DMSO-d6)δ172.79,171.26,142.10,142.00,133.01,131.88,128.09,127.91,127.39,126.23,125.96,125.81 ,125.39,124.42,124.18,123.74,116.33,116.05,64.84,48.15,38.99,35.87,34.34,28.99,28.51,26.21,25.38.ESI-MS m / z: 472.6[MH] - .

[0112] N-(6-(2-aminophenyl)amino)-6-oxohexyl)-4-(naphth-2-yl)tetrahydro-2H-pyran-4-carboxamide (J27)

[0113] J27 was synthesized using the same method as J22. It is a white solid with a yield of 31.8%. Mp: 98-100℃. 1H NMR(600MHz,DMSO-d6)δ9.05(s,1H),7.91(d,J=7.9Hz,1H),7.88-7.81(m,3H),7.68(t,J=5.8Hz,1H),7.54-7.44(m,3 H),7.13(dd,J=7.8,1.6Hz,1H),6.88(td,J=7.6,1.5Hz,1H),6.70(dd,J=8.0,1.5Hz,1H),6.53(td,J=7.6,1.5Hz,1H), 4.80(s,2H),3.78(dt,J=11.9,3.6Hz,2H),3.57-3.47(m,2H),3.03(q,J=6.6Hz,2H),2.54(d,J=13.4Hz,2H),2.21(t,J =7.5Hz,2H),1.94(ddd,J=14.3,10.7,4.0Hz,2H),1.50(p,J=7.5Hz,2H),1.36(p,J=7.2Hz,2H),1.17(p,J=7.7Hz,2H). 13 C NMR(151MHz,DMSO-d6)δ172.67,171.07,141.95,141.86,132.88,131.74,127.97,127.78,127.26,126.10,125.83,125 .67,125.27,124.31,124.07,123.58,116.18,115.89,64.70,48.61,48.02,35.67,34.23,28.79,26.03,24.95.ESI-MS m / z: 458.5[MH] - .

[0114] N-(5-(2-aminophenyl)amino)-5-oxopentyl)-4-(naphth-2-yl)tetrahydro-2H-pyran-4-carboxamide (J28)

[0115] J28 was synthesized using the same method as J22. It is a white solid with a yield of 15.8%. Mp: 194-195℃. 1H NMR(600MHz,DMSO-d6)δ9.07(s,1H),7.92(d,J=7.7Hz,1H),7.87(d,J=10.5Hz,3H),7.75(d,J=5.9Hz,1H),7 .51(dq,J=14.1,8.0,6.8Hz,3H),7.14(d,J=7.9Hz,1H),6.90(t,J=7.6Hz,1H),6.72(d,J=8.0Hz,1H),6.54(t ,J=7.6Hz,1H),4.83(s,2H),3.81(dd,J=11.6,4.3Hz,2H),3.55(t,J=11.1Hz,2H),3.07(q,J=6.6Hz,2H),2. 56(d,J=13.4Hz,2H),2.27(t,J=7.3Hz,2H),2.01-1.89(m,2H),1.48(q,J=7.5Hz,2H),1.40(p,J=7.1Hz,2H). 13 CNMR(151MHz,DMSO-d6)δ172.68,170.99,141.95,141.82,132.86,131.72,127.96,127.77,127.24,126.08,125.81 ,125.64,125.22,124.29,124.04,123.53,116.13,115.85,64.68,48.03,38.76,35.31,34.26,28.65,22.70.ESI-MS m / z: 445.5[MH] - .

[0116] N-((1r,4r)-4-((2-aminophenyl)carbamoyl)cyclohexyl)-4-(naphth-2-yl)tetrahydro-2H-pyran-4-carboxamide (J29)

[0117] J29 was synthesized using the same method as J22. It is a white solid with a yield of 52.6%. Mp: 252-253℃. 1H NMR(600MHz, DMSO-d6)δ9.02(s,1H),7.92(d,J=7.8Hz,1H),7.90-7.82(m,3H),7.55-7.46(m,3H),7.44(d,J=8.1Hz,1H),7 .11(dd,J=7.9,1.5Hz,1H),6.87(td,J=7.6,1.5Hz,1H),6.70(dd,J=8.1,1.4Hz,1H),6.55-6.48(m,1H),4.76(s,2H),3.81 (dt,J=11.7,3.6Hz,2H),3.68-3.57(m,1H),3.52(t,J=11.2Hz,2H),2.56(d,J=13.3Hz,2H),2.24(tt,J=12.1,3.5Hz,1H), 1.98-1.88(m,2H),1.82(d,J=12.3Hz,2H),1.68(dd,J=13.0,4.1Hz,2H),1.44(dt,J=13.2,10.4Hz,2H),1.27-1.14(m,2H). 13 C NMR(151MHz,DMSO-d6)δ173.83,171.88,142.13,141.92,132.88,131.73,127.98,127.76,127.27,126.10,125.83, 125.70,125.32,124.29,123.98,123.47,116.19,115.91,64.73,48.08,47.68,43.46,34.35,31.20,28.41.ESI-MS m / z: 470.6[MH] - .

[0118] N-(2-Aminophenyl)-5-(4-(naphthyl-2-yl)tetrahydro-2H-pyran-4-carboxamine)-1H-pyrazole-3-carboxamide (J30)

[0119] The synthesis method for J30 is the same as that for J22. It is a white solid with a yield of 4.4%. Mp: 231-232℃. 1H NMR(600MHz,DMSO-d6)δ13.22(s,1H),10.15(s,1H),9.62(s,1H),8.04-7.92(m,2H),7 .92-7.85(m,2H),7.58(dt,J=8.7,2.2Hz,1H),7.56-7.47(m,2H),7.17(s,1H),6.95(t ,J=7.7Hz,1H),6.76(d,J=7.8Hz,1H),6.58(t,J=7.6Hz,1H),4.90(s,2H),3.85(d,J=1 1.4Hz, 2H), 3.59 (t, J = 11.2Hz, 2H), 2.75 (d, J = 13.6Hz, 2H), 2.07 (q, J = 9.2, 6.3Hz, 2H). 13 C NMR (151MHz, DMSO-d6) δ171.56,142.73,140.75,132.87,131.86,128.02,127.98,127.30,126.42,126. 35,126.23,126.03,124.67,124.23,116.34,116.27,64.62,48.77,34.00,tautomerizationbroadened signals 5C not seen.ESI-MS m / z: 454.5[MH] - .

[0120] Example 2

[0121] In vitro HDACs inhibitory activity assay of the compound represented by general formula (1)

[0122] Using SAHA and PXD101 as positive controls, and HeLa cell nuclear extract (mainly containing HDAC1 and HDAC2) as the enzyme source, preliminary screening of HDACs enzyme inhibitory activity was performed on all target compounds. The HDAC fluorescence analysis method (two-step method) is rapid, convenient, and simple to operate for detecting HDAC activity. In the first step, the HDAC fluorescent substrate (containing an acetylated lysine side chain -Boc-Lys(acetyl)-AMC) is deacetylated with HDACs enzymes (HeLa cell nuclear extract, mainly containing HDAC1 and HDAC2), activating the substrate and releasing Boc-Lys-AMC. In the second step, Boc-Lys-AMC is hydrolyzed with trypsin to release the fluorescent group AMC, and the fluorescence intensity is measured at the excitation / emission wavelengths (390 nm / 460 nm).

[0123] 1) Add 50 μL of different concentrations of the test compound solution to a 96-well fluorescent plate, with three replicates for each concentration. Add 50 μL of HDAC buffer to both the standard and blank groups. Then add 10 μL of enzyme solution to the experimental and standard groups, and add 10 μL of HDAC buffer to the blank group. After adding all the solutions, place the 96-well fluorescent plate in a 37°C shaker for 5 min.

[0124] 2) After incubation, add 40 μL of the fluorescent substrate Boc-Lys(acetyl)-AMC solution to each well and incubate for 30 min under the same conditions.

[0125] 3) After incubation, add 100 μL of trypsin stop solution to each well and incubate for 20 min under the same conditions. After incubation, measure the fluorescence intensity at wavelengths of 390 nm / 460 nm.

[0126] 4) Calculate the inhibition rate according to the following formula, and fit the inhibition rate of the compound and its corresponding concentration to an S-curve using GraphpadPrism.9 to calculate the IC50. 50 Values. The results are shown in Table 1.

[0127]

[0128] The experimental results show that the enzyme-inhibiting activities of different compounds vary greatly. Among them, compounds J1, J4, J8, J16, and J17 have better enzyme-inhibiting activities, and J8 and J17 have better enzyme-inhibiting activities than the lead compound JT86.

[0129] Example 3

[0130] Experiments on the anti-tumor cell proliferation of some compounds shown in general formula (1)

[0131] Using SAHA and PXD101 as positive controls, the CCK-8 assay was employed to evaluate the antiproliferative activity of some compounds of this invention against cervical cancer cell lines HeLa, colon cancer cell line HCT-116, human non-small cell lung cancer cell line A549, and breast cancer cell lines MDA-MB-231 and MDA-MB-468. Compared with the MTT assay, the CCK-8 assay is less toxic to cells, does not affect cell morphology, and produces a water-soluble product that does not require the addition of DMSO, making it safer and more convenient. Furthermore, the reproducibility of this method is superior to that of the MTT assay. Therefore, the CCK-8 assay was primarily used in this invention to study the antiproliferative activity of the target compounds against tumor cells.

[0132] Cells that have reached the logarithmic growth phase were digested, collected, centrifuged, and then a certain volume of cell culture medium was added and the mixture was pipetted to prepare a cell suspension. The cell suspension was then used for cell counting. All cells used were adherent cells, with 4000-6000 cells per well. Three groups were established: a control group, a standard group, and an experimental group.

[0133] Control group: 100 μL of cell-free complete culture medium was incubated in a 37°C, 5% CO2 cell culture incubator for 24 h. Then, 100 μL of complete culture medium was added, and the cell culture was incubated for another 48 h. The culture medium was then aspirated, and 200 μL of complete culture medium containing 10% CCK-8 was added. The cell culture was then incubated for another 0.5–4 h. The absorbance was measured at 450 nm.

[0134] Standard group: 100 μL of cell suspension was incubated in a 37℃, 5% CO2 cell culture incubator for 24 h, then 100 μL of complete culture medium was added, and the mixture was incubated for another 48 h. The culture medium was then removed, and 200 μL of complete culture medium containing 10% CCK-8 was added, followed by incubation for another 0.5–4 h. The absorbance was measured at 450 nm.

[0135] Experimental group: 100 μL of cell suspension was incubated in a 37℃, 5% CO2 cell culture incubator for 24 h. Then, 100 μL of complete culture medium containing different concentrations of drugs was added, and the mixture was incubated for another 48 h. The culture medium was then removed, and 200 μL of complete culture medium containing 10% CCK-8 was added. The mixture was then incubated for another 0.5–4 h. The absorbance was measured at 450 nm.

[0136] To eliminate edge effects, add 200 μL of PBS buffer to the edge wells.

[0137]

[0138] The inhibition rate (%) of the compound and its corresponding concentration were fitted with an S-curve using Graphpad Prism.9 to calculate the IC50. 50 The values ​​are shown in Table 2.

[0139] Based on a preliminary evaluation of the in vitro HDACs' inhibitory activity, J1, J4, J8, J16, and J17 were selected for anti-tumor cell proliferation experiments in this embodiment. The five compounds with strong inhibitory activity all exhibited higher anti-proliferative activity than the marketed clinical drugs SAHA and PXD101. Compounds J8 and J17 showed superior anti-proliferative activity compared to the lead compound JT86, while compound J8 showed IC50 activity against five types of solid tumors. 50 The values ​​are all less than 100 nM.

[0140]

[0141] Table 1. Evaluation results of the in vitro HDAC inhibitory activity of the compounds.

[0142]

[0143]

[0144]

[0145] a All compounds were tested at least two times, the SD values ​​are<20% of mean.

[0146] Table 2. Evaluation results of the antitumor cell proliferation activity of some compounds.

[0147]

[0148] b IC 50 (nM)are the mean of three independent assays,expressed as mean±standard deviation(SD).

[0149] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. An HDACs inhibitor, characterized in that, The inhibitor is selected from the following compounds: 。 2. A method for preparing an HDACs inhibitor as described in claim 1, characterized in that, The synthetic route for compound J1 is as follows: 0.78 mmol of 4-naphthalene-tetrahydropyran-4-carboxylic acid was placed in a two-necked flask, and 5 mL of thionyl chloride was added. Under nitrogen protection, the mixture was heated to reflux at 80 °C for 4 h. The thionyl chloride was removed by vacuum distillation, and the residue was washed twice with redistilled DCM. The solvent was removed by vacuum distillation, and redistilled DCM was added to prepare solution a. Separately, 0.94 mmol of the product (E)-4-aminocinnamate ethyl ester was dissolved in redistilled DCM and placed in a two-necked flask to prepare solution b. Solution a was added dropwise to solution b, followed by the addition of 2.34 mmol of DIPEA and 0.5 mL of DMF. Under nitrogen protection, the mixture was reacted overnight at room temperature. After the reaction was completed, the mixture was extracted twice each with 2M HCl, saturated NaHCO3 solution, and saturated sodium chloride solution. The extract was dried with an appropriate amount of anhydrous magnesium sulfate, filtered, and the solvent was removed by vacuum distillation to obtain crude product 7a. The structural formula of 7a is as follows: ，R= ； Preparation of NH2OK solution: Dissolve 14.18 g KOH in 35 mL MeOH under ice bath to prepare solution a, and dissolve 11.68 g hydroxylamine hydrochloride in 60 mL MeOH to prepare solution b. Add solution a dropwise to solution b, stir for 1 h, and filter to remove the precipitate to obtain NH2OK solution; Dissolve crude product 7a in 12 mL NH2OK solution and react at room temperature for 30 min; After the reaction is complete, remove the solvent under reduced pressure, add an appropriate amount of water, adjust the pH to 1 with 2M HCl, extract three times with EA, extract the organic phase twice with saturated sodium chloride solution, add an appropriate amount of anhydrous magnesium sulfate to dry, filter, remove the solvent under reduced pressure, and purify by HPLC to obtain yellow solid product J1.

3. The use of the HDACs inhibitor according to claim 1 or the HDACs inhibitor prepared by the method according to claim 2 in the preparation of antitumor drugs.

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