A HDAC11 subtype selective inhibitor and its preparation method and application

Abstract

The present invention provides a selective inhibitor of HDAC11 isoforms, its preparation method, and use. The inhibitors of the present invention have the following general structural formulas: A or B, as well as optical isomers and pharmaceutically acceptable salts thereof. These inhibitors have the potential to combat drug resistance, recurrence, and metastasis in hepatocellular carcinoma and can be used to prepare drugs for the prevention or treatment of diseases associated with abnormal HDAC11 expression or activity. #imgabs0#

Description

Technical Field

[0001] The present invention relates to the technical field of organic compound synthesis and pharmaceutical application, and in particular to an HDAC11 subtype selective inhibitor, a preparation method and application thereof. Background Art

[0002] Histone deacetylase (HDAC) is an important class of epigenetic modification-related proteins. There are 18 subtypes of human HDACs discovered so far, of which HDAC1-11 are zinc ion-dependent metalloproteinases. HDAC11 is the latest zinc ion-dependent HDAC discovered. It is distributed in both the cytoplasm and the nucleus and plays a variety of physiological and pathological functions. It is worth noting that several recent studies have shown that HDAC11 has a very strong de-long-chain fatty acid acylation activity (see: Cao J. et al., Proc Natl Acad Sci USA. 2019, 116, 5487-5492; Kutil Z. et al., ACS Chem. Biol. 2018, 13, 685-693).

[0003] High expression of HDAC11 is closely associated with the occurrence, development, and poor prognosis of various cancers, including hepatocellular carcinoma, myeloproliferative neoplasms, multiple myeloma, Hodgkin's lymphoma, non-small cell lung cancer, glioblastoma, pituitary tumors, prostate cancer, ovarian cancer, and acute lymphoblastic leukemia. Therefore, it is considered a potential target for cancer therapy (see: Liu S. et al., Biomed. Pharmacother, 2020, 131, 110607; Y. et al., FEBS J. https: / / doi.org / 10.1111 / febs.15895). For example, several recent studies have found that the expression level of histone deacetylase 11 (HDAC11) in hepatocellular carcinoma tissues / cells is significantly higher than that in normal liver tissues / cells, and that high HDAC11 expression is closely associated with sorafenib resistance and poor patient prognosis (see: Bi L. et al., Cancer Res. 2021, 81, 2015-2028; Wang W. et al., Front. Cell Dev. Biol. 2020, 8, 724; Freese K. et al., Cancers 2019, 11, 1587; Gong D. et al., Am. J. Transl. Res. 2019, 11, 983-990). Correspondingly, HDAC11 knockout / knockdown can effectively inhibit the growth, invasion and metastasis of hepatocellular carcinoma, reduce the stemness of cancer stem cells (CSCs) and sorafenib resistance (see: Bi L. et al., Cancer Res. 2021, 81, 2015-2028; Wang W. et al., Front. Cell Dev. Biol. 2020, 8, 724). In summary, HDAC11 is a potential target for cancer therapy. Since HDAC11 is crucial for the maintenance of CSC stemness (see: Bi L. et al., Cancer Res. 2021, 81, 2015-2028), and CSCs are closely related to tumor resistance, recurrence and metastasis (see: Lytel N, et al., Nat. Rev. Cancer 2018, 18, 669-680), HDAC11 subtype selective inhibitors are also expected to solve the difficult problems of cancer resistance, recurrence and metastasis in cancer treatment.

[0004] In addition, a large number of studies have shown that HDAC11 selective inhibitors are also expected to be used to treat autoimmune diseases such as inflammation, psoriasis, rheumatoid arthritis, rheumatoid arthritis, and systemic lupus erythematosus, as well as metabolic diseases such as obesity and diabetes (see: Liu S. et al., Biomed. Pharmacother, 2020, 131, 110607; Y. et al., FEBS J. https: / / doi.org / 10.1111 / febs.15895).

[0005] Currently, the research and development of HDAC11 subtype selective inhibitors is in its infancy both domestically and internationally, and only three compounds (FT895, SIS17, and garcinol) with strong HDAC11 selective inhibitory activity have been reported (see: Y.et al.,FEBS J.https: / / doi.org / 10.1111 / febs.15895), and its activity is still insufficiently studied.

[0006] Summary of the Invention

[0007] In view of the deficiencies of the prior art, the present invention provides an HDAC11 subtype selective inhibitor, and also provides a preparation method and application of the compound.

[0008] The technical solutions of the present invention are as follows:

[0009] 1. HDAC11 isoform-selective inhibitors

[0010] Inhibitors having the structure shown in the following general formula A or B, and optical isomers and pharmaceutically acceptable salts thereof:

[0011]

[0012] in,

[0013] R1 is an aromatic ring, an aromatic heterocycle, a substituted aromatic ring or a substituted aromatic heterocycle; wherein the substituent in the substituted aromatic ring or substituted aromatic heterocycle is selected from alkyl, aminoalkyl, halogen, haloalkyl, alkoxy, amino, amine, piperazinyl, alkylpiperazinyl, morpholinyl, alkylmorpholinyl, cyano, alkylamide or amide;

[0014] R2 is hydrogen, alkyl, alkylpiperazinyl, alkylmorpholinyl or aminoalkyl;

[0015] R3 is a hydroxyl group, an amino group or a primary amino group.

[0016] According to the present invention, preferably,

[0017] R1 is a benzene ring, a substituted benzene ring, a pyridine ring, a substituted pyridine ring, a pyrazine ring, a substituted pyrazine ring, a pyrimidine ring or a substituted pyrimidine ring; wherein the substituent of the substituted benzene ring, substituted pyridine ring, substituted pyrazine ring or substituted pyrimidine ring is selected from

[0018] R2 is

[0019] R3 is a hydroxyl group, an amino group, or a primary amino group substituted by an alkyl group having 1 to 16 carbon atoms.

[0020] Preferably, the structure of the inhibitor is one of the following:

[0021]

[0022]

[0023] 2. Preparation of HDAC11 subtype selective inhibitors

[0024] The preparation method of the inhibitor wherein R3 is a hydroxyl group in the general structural formula A of the inhibitor is selected from the following:

[0025] (1) Compound 1 reacts with p-toluenesulfonyl chloride to generate compound 2, compound 2 reacts with carbon tetrabromide to generate compound 3, and compound 3 reacts with Compound 4 is generated by Sonogashira coupling reaction, and compound 4 reacts with potassium hydroxylamine to obtain compounds A1-A13;

[0026] Alternatively, compound 4 is deprotected to generate compound 16, and compound 16 is reacted with The reaction generates compound 17, which reacts with potassium hydroxylamine to obtain compounds A51-A55.

[0027] The reaction formula is as follows:

[0028]

[0029] Wherein, in the reaction formula for preparing compounds A1-A13, the substituent R1 is the same as the substituent R1 in the corresponding compounds A1-A13; in the reaction formula for preparing compounds A51-A55, the substituents R1 and R2 are the same as the substituents R1 and R2 in the corresponding compounds A51-A55;

[0030] Reagents and conditions in the above reaction formula:

[0031] a. p-Toluenesulfonyl chloride, sodium hydride, tetrahydrofuran, reaction at room temperature;

[0032] b. Carbon tetrabromide, lithium diisopropylamide, tetrahydrofuran, reaction at -78°C and room temperature;

[0033] c. Cuprous iodide, triethylamine, bis(triphenylphosphine)palladium dichloride, 1,4-dioxane, reaction at 70°C;

[0034] d. Potassium hydroxylamine, methanol, room temperature reaction;

[0035] e. Tetrabutylammonium fluoride; tetrahydrofuran; reaction at 70°C;

[0036] f. Sodium hydride, tetrahydrofuran, room temperature reaction;

[0037] (2) Compound 1 reacts with di-tert-butyl dicarbonate to produce compound 5, compound 5 reacts with carbon tetrabromide to produce compound 6, compound 6 reacts with trimethylsilylacetylene via Sonogashira coupling to produce compound 7, compound 7 reacts to produce compound 8, compound 8 reacts with R1I via Sonogashira coupling to produce compound 9, compound 9 reacts with potassium hydroxylamine to produce compound 10, and compound 10 is deprotected to produce compounds A14-A20.

[0038] The reaction formula is as follows:

[0039]

[0040] Wherein, the substituent R1 in the reaction formula is the same as the substituent R1 in the corresponding compounds A14-A20;

[0041] Reagents and conditions in the above reaction formula:

[0042] a. Di-tert-butyl dicarbonate, triethylamine, dichloromethane, reaction at room temperature;

[0043] b. Carbon tetrabromide, lithium diisopropylamide, tetrahydrofuran, reaction at -78°C and room temperature;

[0044] c. Cuprous iodide, triethylamine, bis(triphenylphosphine)palladium dichloride, 1,4-dioxane, reaction at 70°C;

[0045] d. Tetrabutylammonium fluoride; tetrahydrofuran; reaction at 70°C;

[0046] e. R1I, cuprous iodide, triethylamine, bis(triphenylphosphine)palladium dichloride, 1,4-dioxane, reaction at 70°C;

[0047] f. Potassium hydroxylamine, methanol, room temperature reaction;

[0048] g. Trifluoroacetic acid, dichloromethane, reaction at room temperature;

[0049] (3) Compound 1 reacts with R2I to produce compound 11, compound 11 reacts with elemental iodine to produce compound 12, compound 12 reacts with trimethylsilylacetylene via Sonogashira coupling to produce compound 13, compound 13 reacts to produce compound 14, compound 14 reacts with R1I via Sonogashira coupling to produce compound 15, and compound 15 reacts with potassium hydroxylamine to produce compounds A21-A50;

[0050] The reaction formula is as follows:

[0051]

[0052] Wherein, the substituents R1 and R2 in the reaction formula are the same as the substituents R1 and R2 in the corresponding compounds A21-A50;

[0053] Reagents and conditions in the above reaction formula:

[0054] a. R2I, sodium hydride, tetrahydrofuran, room temperature reaction;

[0055] b. Iodine, n-butyl lithium, tetrahydrofuran, -78 ° C and room temperature reaction;

[0056] c. Cuprous iodide, triethylamine, bis(triphenylphosphine)palladium dichloride, 1,4-dioxane, reaction at 70°C;

[0057] d. Tetrabutylammonium fluoride; tetrahydrofuran; reaction at 70°C;

[0058] e. R1I, cuprous iodide, triethylamine, bis(triphenylphosphine)palladium dichloride, 1,4-dioxane, reaction at 70°C;

[0059] f. Potassium hydroxylamine, methanol, room temperature reaction;

[0060] The preparation method of the inhibitor wherein R3 in the inhibitor structural formula A is an amino group or a primary amine group is selected from one of the following:

[0061] (iv) Compound 8 reacts with R1I via Sonogashira coupling to generate compound 18, compound 18 is hydrolyzed to generate compound 19, compound 19 is condensed with hydrazine hydrate to generate compound 20, and compound 20 is deprotected to generate compound A56;

[0062] Alternatively, compound 20 undergoes reductive amination with a fatty aldehyde to generate compound 21, and compound 21 is deprotected to generate compound A57;

[0063] The reaction formula is as follows:

[0064]

[0065] The substituent R1 in the reaction formula is the same as the substituent R1 in compound A56 and compound A57, which is phenyl; and in compound 21 and compound A57, n is 2.

[0066] Reagents and conditions in the above reaction formula:

[0067] a. R1I, cuprous iodide, triethylamine, bis(triphenylphosphine)palladium dichloride, 1,4-dioxane, 70°C;

[0068] b. Sodium hydroxide, methanol, reaction at 50°C;

[0069] c. Hydrazine hydrate, O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroboric acid, triethylamine, dimethyl sulfoxide, reaction at room temperature;

[0070] d. Trifluoroacetic acid, dichloromethane, reaction at room temperature;

[0071] e. Fatty aldehyde, sodium cyanoborohydride, methanol, glacial acetic acid, reaction at room temperature;

[0072] (5) Compound 15 is hydrolyzed to generate compound 22, and compound 22 is condensed with hydrazine hydrate to obtain compound A58;

[0073] Alternatively, compound A58 undergoes reductive amination reaction with aliphatic aldehydes to produce compounds A59-A61.

[0074] The reaction formula is as follows:

[0075]

[0076] Wherein, the substituents R1 and R2 in the reaction formula are the same as the substituents R1 and R2 in compounds A58-A61; in compound A59, n is 2; in compound A60, n is 15; in compound A61, n is 2;

[0077] Reagents and conditions in the above reaction formula:

[0078] a. Sodium hydroxide, methanol, 50℃ reaction;

[0079] b. Hydrazine hydrate, O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroboric acid, triethylamine, dimethyl sulfoxide, reaction at room temperature;

[0080] c. Fatty aldehyde, sodium cyanoborohydride, methanol, glacial acetic acid, reaction at room temperature;

[0081] The preparation method of the inhibitor wherein R3 is a hydroxyl group in the general structural formula B is selected from the following:

[0082] (VI) Compound 23 reacts with p-toluenesulfonyl chloride to generate compound 24, which reacts with carbon tetrabromide to generate compound 25, which reacts with Compound 26 is generated by Sonogashira coupling reaction, and compound 26 reacts with potassium hydroxylamine to obtain compounds B1-B3.

[0083] The reaction formula is as follows:

[0084]

[0085] Wherein, the substituent R1 in the reaction formula is the same as the substituent R1 in the corresponding compounds B1-B3;

[0086] Reagents and conditions in the above reaction formula:

[0087] a. p-Toluenesulfonyl chloride, sodium hydride, tetrahydrofuran, reaction at room temperature;

[0088] b. Carbon tetrabromide, lithium diisopropylamide, tetrahydrofuran, reaction at -78°C and room temperature;

[0089] c. Cuprous iodide, triethylamine, bis(triphenylphosphine)palladium dichloride, 1,4-dioxane, reaction at 70°C;

[0090] d. Potassium hydroxylamine, methanol, room temperature reaction;

[0091] (VII) Compound 23 reacts with di-tert-butyl dicarbonate to produce compound 27, compound 27 reacts with carbon tetrabromide to produce compound 28, compound 28 reacts with trimethylsilylacetylene via Sonogashira coupling to produce compound 29, compound 29 reacts to produce compound 30, compound 30 reacts with R1I via Sonogashira coupling to produce compound 31, compound 31 reacts with potassium hydroxylamine to produce compound 32, and compound 32 is deprotected to produce compounds B4-B6.

[0092] The reaction formula is as follows:

[0093]

[0094] Wherein, the substituent R1 in the reaction formula is the same as the substituent R1 in the corresponding compounds B4-B6;

[0095] Reagents and conditions in the above reaction formula:

[0096] a. Di-tert-butyl dicarbonate, triethylamine, dichloromethane, reaction at room temperature;

[0097] b. Carbon tetrabromide, lithium diisopropylamide, tetrahydrofuran, reaction at -78°C and room temperature;

[0098] c. Cuprous iodide, triethylamine, bis(triphenylphosphine)palladium dichloride, 1,4-dioxane, reaction at 70°C;

[0099] d. Tetrabutylammonium fluoride; tetrahydrofuran; reaction at 70°C;

[0100] e. R1I, cuprous iodide, triethylamine, bis(triphenylphosphine)palladium dichloride, 1,4-dioxane, reaction at 70°C;

[0101] f. Potassium hydroxylamine, methanol, room temperature reaction;

[0102] g. Trifluoroacetic acid, dichloromethane, reaction at room temperature;

[0103] (VIII) Compound 23 reacts with R2I to generate compound 33, compound 33 reacts with elemental iodine to generate compound 34, compound 34 reacts with trimethylsilylacetylene via Sonogashira coupling to generate compound 35, compound 35 reacts to generate compound 36, compound 36 reacts with R1I via Sonogashira coupling to generate compound 37, and compound 37 reacts with potassium hydroxylamine to obtain compound B7.

[0104] The reaction formula is as follows:

[0105]

[0106] Wherein, the substituents R1 and R2 in the reaction formula are the same as the substituents R1 and R2 in the corresponding compound B7;

[0107] Reagents and conditions in the above reaction formula:

[0108] a. R2I, sodium hydride, tetrahydrofuran, room temperature reaction;

[0109] b. Iodine, n-butyl lithium, tetrahydrofuran, -78 ° C and room temperature reaction;

[0110] c. Cuprous iodide, triethylamine, bis(triphenylphosphine)palladium dichloride, 1,4-dioxane, reaction at 70°C;

[0111] d. Tetrabutylammonium fluoride; tetrahydrofuran; reaction at 70°C;

[0112] e. R1I, cuprous iodide, triethylamine, bis(triphenylphosphine)palladium dichloride, 1,4-dioxane, reaction at 70°C;

[0113] f. Potassium hydroxylamine, methanol, room temperature reaction;

[0114] 3. Application of HDAC11 isoform-selective inhibitors

[0115] Use of an HDAC11 subtype selective inhibitor in the preparation of a drug for preventing or treating diseases associated with abnormal HDAC11 expression or activity.

[0116] The disease associated with abnormal HDAC11 expression or activity is cancer, autoimmune disease or metabolic disease.

[0117] The cancer is liver cancer, myeloproliferative neoplasms, multiple myeloma, Hodgkin's lymphoma, non-small cell lung cancer, glioblastoma, pituitary tumor, prostate cancer, ovarian cancer or acute lymphoblastic leukemia.

[0118] The autoimmune disease is inflammation, psoriasis, rheumatoid arthritis, rheumatoid arthritis or systemic lupus erythematosus.

[0119] The metabolic disease is obesity or diabetes.

[0120] A pharmaceutical composition for preventing or treating cancer, autoimmune disease or metabolic disease comprises the HDAC11 subtype selective inhibitor of the present invention or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers or excipients.

[0121] The beneficial effects of the present invention are as follows:

[0122] The HDAC11 subtype-selective inhibitor of the present invention possesses a novel structure and, compared to existing inhibitors, has potential advantages in combating drug resistance, recurrence, and metastasis in hepatocellular carcinoma. The HDAC11 subtype-selective inhibitor of the present invention exhibits superior inhibitory activity and subtype selectivity for HDAC11, exhibits moderate antiproliferative activity against tumor cells, and exhibits low toxicity against normal cells. It can inhibit hepatocellular carcinoma cell colony formation and tumor stem cell spheroidization, and effectively inhibits hepatocellular carcinoma cell migration. BRIEF DESCRIPTION OF THE DRAWINGS

[0123] Figure 1 Inhibitory activity of compound A1 and sorafenib on Huh7 cell colony formation in Experimental Example 3;

[0124] Figure 2 Inhibitory activity of compound A1 and sorafenib on Huh7 tumor stem cell spheroid formation in Experimental Example 3;

[0125] Figure 3 . Inhibitory activity of compound A1 and sorafenib in Experimental Example 4 on Huh7 cell migration. DETAILED DESCRIPTION

[0126] The present invention will be further described below with reference to the embodiments, but is not limited thereto.

[0127] Meanwhile, the reagents used in the examples can be purchased commercially unless otherwise specified; the methods and equipment used can be based on existing technologies unless otherwise specified.

[0128] Example 1. Preparation of compounds A1-A13 and B1-B3, taking compound A1 as an example.

[0129] Synthesis route:

[0130]

[0131] The specific synthesis method and steps are as follows:

[0132] Compound 2: NaH (150 mg, 60 wt%) was added to a flask, and then 5 mL of THF was added to the flask at 0 ° C. Compound 1 was dissolved in 10 mL of THF, and then compound 1 (1 g, 5.7 mmol) was added to the stirred solution of NaH via a syringe. After 30 minutes, TsCl (p-toluenesulfonyl chloride, 1.19 g, 6.2 mmol) was added to the solution. The resulting mixture was stirred at 20 ° C for 6 hours. After the reaction was completed, 100 mL of ice water was added thereto, and the precipitated solid was filtered. After drying, EtOAc / PE (1 / 5, 20 mL) was further slurried to obtain a white solid 2 (1.76 g, yield 94%). The NMR data of the product are as follows: 1 H NMR (400MHz, DMSO-d6) δ8.23(d,J=8.3Hz,1H),8.01(d,J=3.7Hz,1H),7.90(dd,J=7.6,5.4Hz,3H ),7.48(t,J=8.0Hz,1H),7.39(d,J=8.2Hz,2H),7.29(d,J=3.6Hz,1H),3.89(s,3H),2.32(s,3H). ESI-MS,m / z=330.3[M+H] + .

[0133] Compound 3: LDA (lithium diisopropylamide, 1.2 mL, 2M THF solution) was added to a THF solution (5 mL) of compound 2 (660 mg, 2 mmol) at -78°C. After 30 minutes, a THF solution (5 mL) of CBr4 (797 mg, 2.4 mmol) was added dropwise. After reacting at this temperature for 30 minutes, the mixture was moved to room temperature and reacted for 30 minutes. After completion of the reaction, the mixture was extracted with ethyl acetate (50 mL) and HCl (0.5 M, 40 mL). The organic layer was washed with brine and dried over MgSO4. The residue was then concentrated to obtain a residue, which was subjected to column chromatography. Petroleum ether / ethyl acetate (volume ratio: 100 / 10) was used to obtain a white solid 3 (130 mg, yield 16%). The NMR data of the product are as follows: 1 H NMR (400MHz, DMSO-d6) δ8.46(d,J=8.5Hz,1H),7.93(d,J=7.7Hz,1H),7.81-7. 76(m,2H),7.52(t,J=8.1Hz,1H),7.47-7.40(m,3H),3.88(s,3H),2.34(s,3H).

[0134] Compound 4a: Compound 3 (340 mg, 0.8 mmol), CuI (32 mg, 0.2 eq), and Pd(PPh3)2Cl2 (bis(triphenylphosphine)palladium dichloride, 130 mg, 0.2 eq) were dissolved in 1,4-dioxane (4 mL). After replacing the argon atmosphere, Et3N (1.5 mL) and ethynylbenzene (102 mg, 1 mmol) were added. The resulting mixture was stirred at 70°C for 20 hours. The reaction solution was concentrated and purified on a silica gel column (PE / EA = 5 / 1) to obtain a brown oil 4a (280 mg, 82% yield). The NMR data of the product are as follows: 1 H NMR (400MHz, DMSO-d6) δ8.46(d,J=8.5Hz,1H),7.93(d,J=7.6Hz,2H),7.79(d,J=8.4Hz,2H),7.77- 7.58(m,1H),7.52(t,J=8.1Hz,2H),7.45(s,2H),7.42(d,J=8.2Hz,2H),3.88(s,3H),2.34(s,3H). ESI-MS,m / z=428.3[MH] - .

[0135] Target compound A1: Weigh hydroxylamine hydrochloride (5.0 g, 72 mmol) and dissolve it in 12 mL of anhydrous methanol. Stir in an ice bath. Weigh KOH (6.06 g, 108 mmol) and add it to 20 mL of anhydrous methanol. Stir until dissolved. Add the KOH solution dropwise to the hydroxylamine hydrochloride solution in an ice bath and continue stirring for 1 hour. Filter to obtain a potassium hydroxylamine solution. Dissolve compound 4a (180 mg, 0.42 mmol) in potassium hydroxylamine solution (6 mL) and stir at room temperature for 10 hours. After the reaction is complete, spin dry the liquid, add water to dissolve it evenly, add 1N hydrochloric acid dropwise to adjust the pH to neutral, precipitate the solid, and filter to obtain the crude product. Purify by reverse phase column chromatography (MeCN / H2O=50% / 50%, HAc conditions) to obtain a light yellow solid A1 (90 mg, 50% yield). The NMR data of the product are as follows: 1 H NMR(400MHz,DMSO-d6)δ11.92(s,1H),11.02(s,1H),8.99(s,1H),7.64-7.56(m,2 H),7.52-7.43(m,4H),7.36(d,J=7.2Hz,1H),7.21(t,J=7.7Hz,1H),7.13(s,1H). ESI-MS,m / z=277.1[M+H] + .

[0136] The preparation methods of compounds A2-A13 and B1-B3 are similar to that of compound A1.

[0137] Example 2. Preparation of compounds A14-A20, B4-B6, taking compound A17 as an example.

[0138] Synthesis route:

[0139]

[0140] The specific synthesis method and steps are as follows:

[0141] Compound 5: Compound 1 (1 g, 5.71 mmol) was dissolved in 20 mL of dichloromethane (dichloromethane), and triethylamine (1 mL) and di-tert-butyl dicarbonate (1.49 g, 6.85 mmol) were added. The mixture was stirred at room temperature for 5 h. After the reaction, column chromatography was performed using petroleum ether / ethyl acetate (volume ratio 100 / 1) to obtain 5 (1.4 g, 89% yield) as a clear oil. The NMR data of the product are as follows: 1 H NMR (400MHz, DMSO-d6) δ8.36 (d, J = 8.2 Hz, 1H), 7.94-7.83 (m, 2H), 7.45 (t, J = 8.0 Hz, 1H), 7.20 (d, J = 3.7 Hz, 1H), 3.92 (s, 3H), 1.64 (s, 9H). ESI-MS,m / z=310.1[M+Cl] - .

[0142] Compound 6: Compound 5 (0.24 g, 0.87 mmol) was dissolved in 5 mL of ultra-dry THF. LDA (0.65 mL, 2 M THF solution) was added to the solution of compound 5 at -78°C. After reacting at this temperature for 30 minutes, a THF solution of CBr4 (0.36 g, 1.1 mmol) (5 mL) was added dropwise. After reacting at this temperature for 30 minutes, the mixture was moved to room temperature and continued to react for 30 minutes. After completion of the reaction, the mixture was extracted with ethyl acetate and HCl (0.5 M). The organic layer was washed with brine and dried over MgSO4. The residue was then concentrated to obtain a residue. The residue was chromatographed on a petroleum ether / ethyl acetate (volume ratio 100 / 10) column to obtain a light yellow oil 6 (0.1 g, yield 33%). The NMR data of the product are as follows: 1 H NMR (400MHz, DMSO-d6) δ8.27(d,J=8.4Hz,1H),7.90(d,J=7.6Hz,1H),7.45(t,J=8.0Hz,1H),7.39(s,1H),3.92(s,3H),1.67(s,9H).

[0143] Compound 7: Compound 6 (100 mg, 0.28 mmol), CuI (11 mg, 0.05 mmol), and Pd(PPh3)2Cl2 (35 mg, 0.05 mmol) were dissolved in 1,4-dioxane (4 mL). After replacing the argon atmosphere, Et3N (2 mL) and trimethylsilylacetylene (41 mg, 0.42 mmol) were added. The resulting mixture was stirred at 70°C for 20 hours. The reaction solution was concentrated and purified by column chromatography on a silica gel column (PE / EA = 5 / 1) to obtain 7 (70 mg, 67% yield) as a brown oil. The NMR data of the product are as follows: 1 H NMR (400MHz, DMSO-d6) δ8.35(d,J=8.4Hz,1H),7.91(d,J=7.6Hz,1H),7.53-7.48(m,2H),3.91(s,3H),1.64(s,9H),0.26(s,9H).

[0144] Compound 8: Compound 7 (170 mg, 0.45 mmol) was dissolved in THF (5 mL), and TBAF (tetrabutylammonium fluoride, 0.18 g, 0.69 mmol) was added. The mixture was stirred at 70°C for 4 hours. After completion of the reaction, column chromatography was performed using petroleum ether / ethyl acetate (volume ratio 100 / 1) to obtain solid 8 (92 mg, 68% yield). The NMR data of the product are as follows: 1 H NMR (400MHz, DMSO-d6) δ8.41(d,J=8.4Hz,1H),7.99(d,J=7.6Hz,1H),7.62-7.55(m,2H),4.86(s,1H),3.98(s,3H),1.71(s,9H).

[0145] Compound 9a: 1-(4-iodophenyl)-N,N-dimethylmethanamine (73 mg, 0.28 mmol), CuI (11 mg, 0.05 mmol), and Pd(PPh3)2Cl2 (35 mg, 0.05 mmol) were dissolved in 1,4-dioxane (4 mL). After purging the atmosphere with argon, Et3N (2 mL) and compound 8 (125 mg, 0.42 mmol) were added. The resulting mixture was stirred at 70°C for 20 hours. The reaction mixture was concentrated and purified by silica gel column chromatography (PE / EA = 5 / 1) to obtain 9a as a brown oil (67 mg, 55% yield). ESI-MS, m / z = 433.4 [M+H] + .

[0146] Compound 10a: Weigh hydroxylamine hydrochloride (5.0 g, 72 mmol) and dissolve it in 12 mL of anhydrous methanol, stirring in an ice bath. Weigh KOH (6.06 g, 108 mmol) and add it to 20 mL of anhydrous methanol and stir until dissolved. Add the KOH solution dropwise to the hydroxylamine hydrochloride solution under ice bath conditions and continue stirring for 1 hour. Filter to obtain a potassium hydroxylamine solution. Dissolve compound 9a (130 mg, 0.30 mmol) in potassium hydroxylamine solution (6 mL) and stir at room temperature for 10 hours. After the reaction is completed, spin dry the liquid, add water to dissolve it evenly, add 1N hydrochloric acid dropwise to adjust the pH until the solid precipitates completely, and filter to obtain the crude product. Purify by reverse phase column chromatography (MeCN / H2O=50% / 50%, HAc conditions) to obtain a light yellow solid 10a (87 mg, yield 67%). The NMR data of the product are as follows: 1 H NMR (400MHz, DMSO-d6) δ11.02(s,1H),8.97(s,1H),7.76-7.23(m,8H),3.39(s,2H),2.13(s,6H),1.63(s,9H).

[0147] Target compound A17: To a CH2Cl2 solution (5 mL) of compound 10a (434 mg, 1.0 mmol) was added 2 mL of TFA (trifluoroacetic acid) and stirred at room temperature for 3 hours. After completion of the reaction, the reaction solution was washed three times with saturated Na2CO3 solution and then concentrated to obtain a residue. The residue was purified by column chromatography with petroleum ether / ethyl acetate (volume ratio 100 / 10) to obtain A17 as a pale yellow solid (217 mg, 65% yield). The NMR data of the product are as follows: 1 H NMR (400MHz, DMSO-d6) δ11.92(s,1H),11.02(s,1H),8.97(s,1H),7.64-7.15(m,8H),3.37(s,2H),2.15(s,6H). ESI-MS,m / z=334.3[M+H] + .

[0148] The preparation methods of compounds A14-A16, A18-A20, and B4-B6 are similar to that of compound A17.

[0149] Example 3. Preparation of compounds A21-A50 and B7, taking compound A21 as an example.

[0150] Synthesis route:

[0151]

[0152] The specific synthesis method and steps are as follows:

[0153] Compound 11a: Compound 1 (1.0 g, 5.7 mmol) was dissolved in THF (5 mL). NaH (150 mg, 60 wt%) was slowly added under ice-cooling. After stirring at 0°C for 30 minutes, CH₃I (1.2 g, 8.4 mmol) was added dropwise. The mixture was allowed to react at room temperature for 5 hours. After completion, the reaction was quenched with NH₃Cl, extracted with ethyl acetate, and the organic phases were combined, dried over anhydrous magnesium sulfate, and concentrated to afford 11a as a colorless, transparent oil (0.82 g, 77% yield). The NMR data of the product are as follows: 1 H NMR (400MHz, DMSO-d6) δ7.77(dd,J=7.7,3.7Hz,2H),7.53(d,J=2.9Hz,1H),7.27(t,J=7.8Hz,1H),6.93(d,J=2.5Hz,1H),3.90(s,3H),3.85(s,3H). ESI-MS,m / z=378.6[2M+H] + .

[0154] Compound 12a: Compound 11a (0.27 g, 0.85 mmol) was dissolved in 10 mL of ultra-dry THF. n-BuLi (0.4 mL, 2.5 M in hexane) was added to the solution at -78°C. After reacting at this temperature for 30 minutes, a solution of iodine (0.24 g, 0.95 mmol) in 5 mL of THF was added. The reaction was continued at this temperature for 30 minutes, then the mixture was brought to room temperature and allowed to react for another 2 hours. After completion, the reaction was quenched with saturated ammonium chloride solution, extracted with ethyl acetate, and the organic layer was washed with brine, dried over MgSO4, and concentrated to afford crude compound 12a, which was used in the next reaction.

[0155] Compound 13a: Crude compound 12a (160 mg), CuI (20 mg, 0.11 mmol), and Pd(PPh3)2Cl2 (77 mg, 0.11 mmol) were dissolved in 1,4-dioxane (4 mL). After replacing the argon atmosphere, Et3N (2 mL) and trimethylsilylacetylene (62 mg, 0.63 mmol) were added. The resulting mixture was stirred at 70°C for 20 hours. The reaction solution was concentrated and purified on a silica gel column (PE / EA = 10 / 1) to obtain 13a (100 mg) as a brown oil. The NMR data of the product are as follows: 1 H NMR (400MHz, DMSO-d6) δ7.84(s,1H),7.66(d,J=8.2Hz,1H),7.43(d,J=7.3Hz,1H),7.24(t,J=7.8Hz,1H),3.78(s,3H),3.76(s,3H),0.15(s,9H). ESI-MS,m / z=286.1[M+H] + .

[0156] Compound 14a: Compound 13a (131 mg, 0.46 mmol) was dissolved in THF (5 mL), and TBAF (0.18 g, 0.69 mmol) was added. The mixture was stirred at 70°C for 4 hours. After completion of the reaction, column chromatography was performed using petroleum ether / ethyl acetate (volume ratio 100 / 1) to obtain solid 14a (69 mg, 70% yield). The NMR data of the product are as follows: 1 H NMR (400MHz, DMSO-d6) δ7.88(s,1H),7.74(d,J=8.2Hz,1H),7.50(d,J=7.3Hz,1H),7.31(t,J=7.8Hz,1H),3.97(s,1H),3.85(s,3H),3.84(s,3H).

[0157] Compound 15a: Iodobenzene (108 mg, 0.53 mmol), CuI (20 mg, 0.11 mmol), and Pd(PPh3)2Cl2 (77 mg, 0.11 mmol) were dissolved in 1,4-dioxane (4 mL). After argon was replaced, Et3N (2 mL) and compound 14a (134 mg, 0.63 mmol) were added. The resulting mixture was stirred at 70°C for 20 hours. The reaction solution was concentrated and purified on a silica gel column (PE / EA = 10 / 1) to obtain a brown oil 15a (101 mg, 66% yield). The NMR data of the product are as follows: 1 H NMR (400MHz, DMSO-d6) δ7.85-7.80 (m, 2H), 7.68 (dd, J = 6.1Hz, 2.7Hz, 2H), 7.5 2-7.47(m,3H),7.38(t,J=7.8Hz,1H),7.33(s,1H),3.94(s,3H),3.93(s,3H).

[0158] Target compound A21: Weigh hydroxylamine hydrochloride (5.00 g, 72 mmol) and dissolve it in 12 mL of anhydrous methanol. Stir in an ice bath. Weigh KOH (6.06 g, 108 mmol) and add it to 20 mL of anhydrous methanol. Stir until dissolved. Add the KOH solution dropwise to the hydroxylamine hydrochloride solution in an ice bath and continue stirring for 1 hour. Filter to obtain a potassium hydroxylamine solution. Dissolve compound 15a (90 mg, 0.31 mmol) in potassium hydroxylamine solution (6 mL) and stir at room temperature for 10 hours. After the reaction is complete, spin dry the liquid, add water to dissolve it evenly, and add 1N hydrochloric acid dropwise to adjust the pH to neutral to precipitate a solid. Filter to obtain the crude product. Purify by reverse phase column chromatography (MeCN / H2O = 50% / 50%, HAc conditions) to obtain a light yellow solid A21 (60 mg, 67% yield). The NMR data of the product are as follows: 1H NMR (400MHz, DMSO-d6) δ11.08(s,1H),9.04(s,1H),7.70-7.64(m,3H),7.51-7. 46(m,3H),7.41(d,J=7.2Hz,1H),7.33-7.27(m,1H),7.19(s,1H),3.91(s,3H). ESI-MS,m / z=291.5[M+H] + .

[0159] The preparation methods of compounds A22-A50 and B7 are similar to that of compound A21.

[0160] Example 4. Preparation of Compounds A51-A55, taking Compound A51 as an example.

[0161] Synthesis route:

[0162]

[0163] The specific synthesis method and steps are as follows:

[0164] Compound 16a: Compound 4a (0.2 g, 0.46 mmol) was dissolved in THF (5 mL), and TBAF (0.18 g, 0.69 mmol) was added. The mixture was stirred at 70°C for 4 hours. After completion of the reaction, column chromatography was performed using petroleum ether / ethyl acetate (volume ratio 100 / 1) to afford 16a as a white solid (90 mg, 70% yield). The NMR data of the product are as follows: 1 H NMR (400MHz, DMSO-d6) δ 12.06 (s, 1H), 7.75 (d, J = 7.3Hz, 1H), 7.55-7.42 (m, 3H), 7.50-7.44 (m, 3H), 7.34-7.22 (m, 2H), 3.84 (s, 3H).

[0165] Compound 17a: Compound 16a (0.55 g, 2.0 mmol) was dissolved in THF (10 mL). NaH (55 mg, 60 wt%) was slowly added under ice-cooling. After stirring at 0°C for 30 minutes, a solution of 2-(dimethylamino)ethyl 4-methylbenzenesulfonate (0.61 g, 2.5 mmol) in 8 mL of THF was added dropwise. The mixture was allowed to react at room temperature for 8 hours. After completion, the reaction was quenched with NH4Cl and extracted with ethyl acetate. The combined organic phases were dried over anhydrous magnesium sulfate, concentrated, and purified on a silica gel column (PE / EA = 10 / 1) to obtain an oily product, 17a (0.52 g, 75%), which was then used for later use. ESI-MS, m / z = 347.3 [M+H] + .

[0166] Target compound A51: Weigh hydroxylamine hydrochloride (5.00 g, 72 mmol) and dissolve it in 12 mL of anhydrous methanol. Stir in an ice bath. Weigh KOH (6.06 g, 108 mmol) and add it to 20 mL of anhydrous methanol. Stir until dissolved. Add the KOH solution dropwise to the hydroxylamine hydrochloride solution in an ice bath and continue stirring for 1 hour. Filter to obtain a potassium hydroxylamine solution. Dissolve compound 17a (104 mg, 0.30 mmol) in potassium hydroxylamine solution (6 mL) and stir at room temperature for 10 hours. After the reaction is complete, spin dry the liquid, add water to dissolve it evenly, and add 1N hydrochloric acid dropwise to adjust the pH until the solid precipitates completely. Filter to obtain the crude product. Purify by reverse phase column chromatography (MeCN / H2O = 50% / 50%, HAc conditions) to obtain a light yellow solid A51 (64 mg, 61% yield). The NMR data of the product are as follows: 1 H NMR (400MHz, DMSO-d6) δ11.02(s,1H),8.98(s,1H),7.78(d,J=7.4Hz,1H),7.54-7.47(m,3H ),7.51-7.44(m,3H),7.30-7.24(m,2H),4.57-4.50(m,2H),2.75-2.69(m,2H),2.90(s,6H). ESI-MS,m / z=348.6[M+H] + .

[0167] The preparation methods of compounds A52-A55 are similar to that of compound A51.

[0168] Example 5. Preparation of Compound A56.

[0169] Synthesis route:

[0170]

[0171] The specific synthesis method and steps are as follows:

[0172] Compound 18a: Iodobenzene (0.11 g, 0.53 mmol), CuI (20 mg, 0.11 mmol), and Pd(PPh3)2Cl2 (77 mg, 0.11 mmol) were dissolved in 1,4-dioxane (4 mL). After replacing the argon atmosphere, Et3N (2 mL) and compound 8 (189 mg, 0.63 mmol) were added. The resulting mixture was stirred at 70°C for 20 hours. The reaction solution was concentrated and purified on a silica gel column (PE / EA = 10 / 1) to obtain a brown oil 18a (139 mg, 70% yield). The NMR data of the product are as follows: 1H NMR (400MHz, DMSO-d6) δ8.39(d,J=8.4Hz,1H),7.95(d,J=7.5Hz,1H),7.66-7. 60(m,3H),7.54(d,J=8.0Hz,1H),7.51-7.45(m,3H),3.94(s,3H),1.65(s,9H).

[0173] Compound 19a: Compound 18a (0.42 g, 1.12 mmol) was dissolved in methanol, 2.5 N NaOH (5 mL) was added, and the mixture was heated to 50°C for 12 hours. After the reaction, the methanol was evaporated, and the pH was adjusted with 1 N HCl until the solid precipitated completely. Filtering afforded a yellow solid, 19a (0.24 g, 60% yield). The NMR data of the product are as follows: 1 H NMR (400MHz, DMSO-d6) δ 12.70 (s, 1H), 7.76 (d, J = 7.4Hz, 1H), 7.51-7.49 (m, 3H), 7.51-7.44 (m, 3H), 7.30-7.23 (m, 2H), 1.64 (s, 9H).

[0174] Compound 20a: Compound 19a (0.23 g, 0.65 mmol) was dissolved in DMSO (5 mL). TBTU (O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroborate, 0.25 g, 0.78 mmol) and TEA (triethylamine, 180 μL) were added under ice-cooling. After 30 minutes, 80 wt% hydrazine hydrate was added and the mixture was allowed to react at room temperature for 3 hours. After completion of the reaction, water was added to precipitate the solid, which was filtered to obtain a yellow solid 20a (0.11 g, 45% yield). The NMR data of the product are as follows: 1 H NMR (400MHz, DMSO-d6) δ 9.63 (s, 1H), 7.76 (d, J = 7.4Hz, 1H), 7.58-7.48 (m, 3H), 7.52-7.43 (m, 3H), 7.32-7.23 (m, 2H), 4.42 (s, 2H), 1.65 (s, 9H).

[0175] Target compound A56: To a CH2Cl2 solution (5 mL) of compound 20a (375 mg, 1.0 mmol) was added 2 mL of TFA and stirred at room temperature for 3 hours. After completion of the reaction, the reaction solution was washed three times with saturated Na2CO3 solution and then concentrated to obtain a residue. The residue was purified by column chromatography with petroleum ether / ethyl acetate (volume ratio 100 / 10) to obtain A56 as a pale yellow solid (220 mg, 80% yield). The NMR data of the product are as follows: 1H NMR (400MHz, DMSO-d6) δ11.93(s,1H),9.60(s,1H),7.61-7.23(m,7H),7.21-7.20(m,2H),4.75(s,2H). ESI-MS,m / z=276.2[M+H] + .

[0176] Example 6. Preparation of Compound A57.

[0177] Synthesis route:

[0178]

[0179] The specific synthesis method and steps are as follows:

[0180] Compound 21a: Compound 20a (108.9 mg, 0.29 mmol) was dissolved in 15 mL of methanol. One drop of glacial acetic acid was added. After 5 minutes, propionaldehyde (17 mg, 0.29 mmol) was added. One hour later, NaCNBH3 (55 mg, 0.87 mmol) was added. The mixture was stirred at room temperature for 2 hours. Purification by reverse-phase column chromatography (CH3OH / H2O = 90% / 10%, HAc) afforded 21a as a pale yellow solid (66 mg, 55% yield). The NMR data of the product are as follows: 1 H NMR(400MHz, DMSO-d6)δ9.50(s,1H),7.60(dd,J=6.6,3.0Hz,2H),7.52-7.45(m,4H),7.36(d,J=7.3Hz,1H),7.22(t,J= 7.7Hz,1H),7.07-7.01(m,1H),5.08(s,1H),2.77(t,J=7.2Hz,2H),1.36-1.45(m,2H),1.62(s,9H),0.85-0.90(m,3H).

[0181] Target compound A57: To a CH2Cl2 solution (5 mL) of compound 21a (417 mg, 1.0 mmol) was added 2 mL of TFA and stirred at room temperature for 3 hours. After completion of the reaction, the reaction solution was washed three times with saturated Na2CO3 solution and then concentrated to obtain a residue. The residue was purified by column chromatography with petroleum ether / ethyl acetate (100 / 10, by volume) to afford A57 as a pale yellow solid (238 mg, 75% yield). The NMR data of the product are as follows: 1H NMR (400MHz, DMSO-d6) δ11.90(s,1H),9.51(s,1H),7.75(d,J=7.4Hz,1H),7.59(s,3H),7.52-7.43( m,3H),7.32-7.23(m,2H),5.05(s,1H),2.76(t,J=7.2Hz,2H),1.43-1.51(m,2H),0.85-0.91(m,3H). ESI-MS,m / z=318.42[M+H] + .

[0182] Example 7. Preparation of Compound A58.

[0183] Synthesis route:

[0184]

[0185] The specific synthesis method and steps are as follows:

[0186] Compound 22a: Compound 15a (0.32 g, 1.12 mmol) was dissolved in 20 mL of methanol, and 2.5 N NaOH (5 mL) was added. The mixture was heated to 50°C and reacted for 12 hours. After the reaction, the methanol was evaporated, and the pH was adjusted with 1 N HCl until the solid precipitated completely. Filtering afforded a yellow solid 22a (0.21 g, 68% yield). The NMR data of the product are as follows: 1 H NMR (400MHz, DMSO-d6) δ12.66(s,1H),7.83(s,1H),7.65-7.60(m,3H),7.41-7.25(m,4H),7.15-7.12(m,1H),3.81(s,3H).

[0187] Target compound A58: Compound 22a (0.18 g, 0.65 mmol) was dissolved in DMSO (5 mL). TBTU (0.25 g, 0.78 mmol) and TEA (180 μL) were added under ice-cooling. After 30 minutes, 80 wt% hydrazine hydrate was added and the mixture was allowed to react at room temperature for 3 hours. After completion of the reaction, water was added to precipitate the solid, which was filtered to obtain A58 as a yellow solid (90 mg, 48% yield). The NMR data of the product are as follows: 1 HNMR (400MHz, DMSO-d6) δ9.49(s,1H),7.82(s,1H),7.65-7.60(m,3H),7.40-7.26(m,4H),7.16-7.13(m,1H),4.49(s,2H),3.85(s,3H). ESI-MS,m / z=290.3[M+H] + .

[0188] Example 8. Preparation of Compounds A59-A61, taking Compound A59 as an example.

[0189] Synthesis route:

[0190]

[0191] The specific synthesis method and steps are as follows:

[0192] Target compound A59: Compound A58 (83.8 mg, 0.29 mmol) was dissolved in 10 mL of methanol. One drop of glacial acetic acid was added. After 5 minutes, propionaldehyde (17 mg, 0.29 mmol) was added. After 1 hour, NaCNBH3 (55 mg, 0.87 mmol) was added. The mixture was stirred at room temperature for 2 hours. Purification by reverse-phase column chromatography (CH3OH / H2O = 90% / 10%, HAc) afforded A59 as a pale yellow solid (53 mg, 55% yield). The NMR data of the product are as follows: 1 H NMR (400MHz, DMSO-d6) δ9.51(s,1H),7.82(s,1H),7.65-7.60(m,3H),7.40-7.26(m,4H),7.16-7. 13(m,1H),5.06(s,1H),3.85(s,3H),2.78(t,J=7.2Hz,2H),1.38-1.53(m,2H),0.83-0.93(m,3H). ESI-MS,m / z=332.4[M+H] + .

[0193] The preparation method of compounds A60-A61 is similar to that of compound A59.

[0194] Experimental Example 1. Evaluation of in vitro HDAC inhibitory activity and subtype selectivity of target compounds

[0195] The present invention uses fluorescence analysis to determine the inhibitory activities of the target compound on HDAC11, HDAC1 and HDAC6, and uses the approved HDAC inhibitor SAHA as a positive control.

[0196] The experimental results (Table 1) show that most of the target compounds of the present invention exhibited inhibition rates exceeding 50% against HDAC11 at a concentration of 0.5 μM, but inhibition rates against HDAC1 and HDAC6 were less than 20%, indicating strong HDAC11 isoform-selective inhibitory activity. SAHA at 0.5 μM exhibited significant inhibition against HDAC1 and HDAC6 (inhibition rates exceeding 50%), but had a weak inhibitory effect on HDAC11 (inhibition rate less than 10%).

[0197] Table 1. In vitro HDAC11, HDAC1 and HDAC6 inhibition test results of the compounds

[0198]

[0199]

[0200] The target compounds A1 and B1 were selected as representatives, and their half-maximal inhibitory concentrations (IC50) on HDAC1, HDAC4, HDAC6, HDAC8 and HDAC11 were measured. 50 The results in Table 2 show that the IC values of A1 and B1 for HDAC11 50 The IC values of HDACs were 0.031 μM and 0.17 μM, respectively, which are much lower than the IC values of other HDAC isoforms. 50 , further confirming that A1 and B1 have good HDAC11 isoform selective inhibitory activity.

[0201] Table 2. IC values of compounds for HDAC1, HDAC4, HDAC6, HDAC8, and HDAC11 in vitro 50 Test results

[0202]

[0203] Experimental Example 2. In vitro anti-proliferation test of target compounds on tumor cells and normal cell toxicity test

[0204] The target compound A1 was selected as a representative and its in vitro antiproliferative activity against human hepatocellular carcinoma cells (Huh7 and PLC / PRF / 5), human erythroleukemia cells (HEL), and mouse myeloma cells P3x63Ag8.653 was evaluated, as well as its cytotoxicity against normal human hepatocytes HL-7702 and human umbilical vein endothelial cells HUVEC. The results in Table 3 show that compound A1 exhibited some antiproliferative activity against several tumor cell lines and had low cytotoxicity against normal cells. It is worth noting that compound A1's activity in inhibiting the proliferation of HCC cells Huh7 and PLC / PRF / 5 was comparable to that of sorafenib, a first-line clinical hepatocellular carcinoma treatment, and exhibited lower cytotoxicity than sorafenib against normal human hepatocytes HL-7702 and human umbilical vein endothelial cells HUVEC.

[0205] Table 3. In vitro anti-proliferation test of the compounds on tumor cells and normal cell toxicity results

[0206]

[0207] Experimental Example 3. In vitro inhibition of hepatocellular carcinoma cell colony formation and tumor stem cell sphere formation by target compounds

[0208] The target compound A1 was selected as a representative to evaluate its inhibitory activity on hepatocellular carcinoma cell colony formation and tumor stem cell spheroid formation in vitro. The results of the colony formation experiment showed that at a concentration of 2 μM, compared with the Ctrl group (DMSO treatment), both compound A1 and sorafenib could effectively inhibit the colony formation of hepatocellular carcinoma cell Huh7, and A1 had a stronger ability to inhibit Huh7 cell colony formation than sorafenib ( Figure 1 ), which may be related to the inhibitory effect of A1 on HCC cancer stem cells. The stem cell sphere formation experiment further confirmed that at a concentration of 2μM, compared with the Ctrl group (DMSO treatment), both compound A1 and sorafenib could effectively inhibit the sphere formation of Huh7 cancer stem cells (primary and secondary sphere formation), and the activity of A1 in inhibiting the sphere formation of Huh7 cancer stem cells was significantly better than that of sorafenib ( Figure 2 Note: The primary sphere formation refers to the sphere formation experiment of Huh7 cells, and the secondary sphere formation refers to the sphere formation experiment of tumor stem cells obtained by digesting tumor stem cell spheres formed by the primary sphere formation of Huh7 cells.

[0209] Experimental Example 4. Inhibition of Hepatocellular Carcinoma Cell Migration by Target Compounds in Vitro (Scratch Test)

[0210] The target compound A1 was selected as a representative to evaluate its inhibitory activity on the migration of hepatocellular carcinoma cells in vitro. Figure 3 ) showed that after treating cells with 2 μM concentration of compounds for 12 h, compared with the Ctrl group (DMSO treatment), both compound A1 and sorafenib could effectively inhibit the migration of Huh7 cells, and A1 had stronger anti-Huh7 cell migration activity than sorafenib.

[0211] The above results suggest that HDAC11 subtype-selective inhibitors have good therapeutic prospects for cancers such as hepatocellular carcinoma, and also have potential advantages in anti-drug resistance, anti-recurrence and anti-metastasis.

Claims

1. An HDAC11 subtype selective inhibitor, characterized in that The inhibitor is an inhibitor having the structure shown in the following general formula A or B, and a pharmaceutically acceptable salt thereof: Wherein, R1 is a benzene ring, a substituted benzene ring, a pyridine ring, or a substituted pyridine ring; wherein the substituents of the substituted benzene ring or the substituted pyridine ring are selected from: R2 is R3 is a hydroxyl group, an amino group, or a primary amino group substituted by an alkyl group having 3 or 16 carbon atoms.

2. The HDAC11 subtype selective inhibitor according to claim 1, characterized in that The inhibitor is selected from one of the following compounds or a pharmaceutically acceptable salt thereof:

3. The method for preparing the HDAC11 subtype selective inhibitor according to claim 2, wherein the method is selected from the following: (1) Compound 1 reacts with p-toluenesulfonyl chloride to generate compound 2, which reacts with carbon tetrabromide to generate compound 3, which reacts with Compound 4 is generated by Sonogashira coupling reaction, and compound 4 reacts with potassium hydroxylamine to obtain compounds A1-A13; Alternatively, compound 4 is deprotected to generate compound 16, and compound 16 is reacted with The reaction generates compound 17, which reacts with potassium hydroxylamine to obtain compounds A51-A55; The reaction formula is as follows: in, In the reaction formulas for preparing compounds A1-A13, the substituent R1 is the same as the substituent R1 in the corresponding compounds A1-A13; in the reaction formulas for preparing compounds A51-A55, the substituents R1 and R2 are the same as the substituents R1 and R2 in the corresponding compounds A51-A55; Reagents and conditions in the above reaction formula: a. p-Toluenesulfonyl chloride, sodium hydride, tetrahydrofuran, reaction at room temperature; b. Carbon tetrabromide, lithium diisopropylamide, tetrahydrofuran, reaction at -78°C and room temperature; c. Cuprous iodide, triethylamine, bis(triphenylphosphine)palladium dichloride, 1,4-dioxane, reaction at 70°C; d. Potassium hydroxylamine, methanol, room temperature reaction; e. Tetrabutylammonium fluoride; tetrahydrofuran; reaction at 70°C; f. Sodium hydride, tetrahydrofuran, room temperature reaction; (ii) Compound 1 reacts with di-tert-butyl dicarbonate to produce compound 5, compound 5 reacts with carbon tetrabromide to produce compound 6, compound 6 reacts with trimethylsilylacetylene via Sonogashira coupling to produce compound 7, compound 7 reacts to produce compound 8, compound 8 reacts with R1I via Sonogashira coupling to produce compound 9, compound 9 reacts with potassium hydroxylamine to produce compound 10, and compound 10 is deprotected to produce compounds A14-A20; The reaction formula is as follows: Wherein, the substituent R1 in the reaction formula is the same as the substituent R1 in the corresponding compounds A14-A20; Reagents and conditions in the above reaction formula: a. Di-tert-butyl dicarbonate, triethylamine, dichloromethane, reaction at room temperature; b. Carbon tetrabromide, lithium diisopropylamide, tetrahydrofuran, reaction at -78°C and room temperature; c. Cuprous iodide, triethylamine, bis(triphenylphosphine)palladium dichloride, 1,4-dioxane, reaction at 70°C; d. Tetrabutylammonium fluoride; tetrahydrofuran; reaction at 70°C; e. R1I, cuprous iodide, triethylamine, bis(triphenylphosphine)palladium dichloride, 1,4-dioxane, reaction at 70°C; f. Potassium hydroxylamine, methanol, room temperature reaction; g. Trifluoroacetic acid, dichloromethane, reaction at room temperature; (3) Compound 1 reacts with R2I to produce compound 11, compound 11 reacts with elemental iodine to produce compound 12, compound 12 reacts with trimethylsilylacetylene via Sonogashira coupling to produce compound 13, compound 13 reacts to produce compound 14, compound 14 reacts with R1I via Sonogashira coupling to produce compound 15, and compound 15 reacts with potassium hydroxylamine to produce compounds A21-A50; The reaction formula is as follows: Wherein, the substituents R1 and R2 in the reaction formula are the same as the substituents R1 and R2 in the corresponding compounds A21-A50; Reagents and conditions in the above reaction formula: a. R2I, sodium hydride, tetrahydrofuran, room temperature reaction; b. Iodine, n-butyl lithium, tetrahydrofuran, -78 ° C and room temperature reaction; c. Cuprous iodide, triethylamine, bis(triphenylphosphine)palladium dichloride, 1,4-dioxane, reaction at 70°C; d. Tetrabutylammonium fluoride; tetrahydrofuran; reaction at 70°C; e. R1I, cuprous iodide, triethylamine, bis(triphenylphosphine)palladium dichloride, 1,4-dioxane, reaction at 70°C; f. Potassium hydroxylamine, methanol, room temperature reaction; (iv) Compound 8 reacts with R1I via Sonogashira coupling to generate compound 18, compound 18 is hydrolyzed to generate compound 19, compound 19 is condensed with hydrazine hydrate to generate compound 20, and compound 20 is deprotected to generate compound A56; Alternatively, compound 20 undergoes reductive amination with a fatty aldehyde to generate compound 21, and compound 21 is deprotected to generate compound A57; The reaction formula is as follows: Wherein, the substituent R1 in the reaction formula is the same as the substituent R1 in compound A56 and compound A57, which is phenyl; in compound 21 and compound A57, n is 2; Reagents and conditions in the above reaction formula: a. R1I, cuprous iodide, triethylamine, bis(triphenylphosphine)palladium dichloride, 1,4-dioxane, 70°C; b. Sodium hydroxide, methanol, reaction at 50°C; c. Hydrazine hydrate, O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroboric acid, triethylamine, dimethyl sulfoxide, reaction at room temperature; d. Trifluoroacetic acid, dichloromethane, reaction at room temperature; e. Fatty aldehyde, sodium cyanoborohydride, methanol, glacial acetic acid, reaction at room temperature; (5) Compound 15 is hydrolyzed to generate compound 22, and compound 22 is condensed with hydrazine hydrate to obtain compound A58; Alternatively, compound A58 undergoes reductive amination with a fatty aldehyde to produce compounds A59-A61; The reaction formula is as follows: Wherein, the substituents R1 and R2 in the reaction formula are the same as the substituents R1 and R2 in compounds A58-A61; in compound A59, n is 2; in compound A60, n is 15; in compound A61, n is 2; Reagents and conditions in the above reaction formula: a. Sodium hydroxide, methanol, 50℃ reaction; b. Hydrazine hydrate, O-benzotriazole-N,N,N',N'-tetramethyluronium tetrafluoroboric acid, triethylamine, dimethyl sulfoxide, reaction at room temperature; c. Fatty aldehyde, sodium cyanoborohydride, methanol, glacial acetic acid, reaction at room temperature; (VI) Compound 23 reacts with p-toluenesulfonyl chloride to generate compound 24, which reacts with carbon tetrabromide to generate compound 25, which reacts with Compound 26 was generated by Sonogashira coupling reaction, and compound 26 was reacted with potassium hydroxylamine to obtain compounds B1-B3; The reaction formula is as follows: Wherein, the substituent R1 in the reaction formula is the same as the substituent R1 in the corresponding compounds B1-B3; Reagents and conditions in the above reaction formula: a. p-Toluenesulfonyl chloride, sodium hydride, tetrahydrofuran, reaction at room temperature; b. Carbon tetrabromide, lithium diisopropylamide, tetrahydrofuran, reaction at -78°C and room temperature; c. Cuprous iodide, triethylamine, bis(triphenylphosphine)palladium dichloride, 1,4-dioxane, reaction at 70°C; d. Potassium hydroxylamine, methanol, room temperature reaction; (VII) Compound 23 reacts with di-tert-butyl dicarbonate to produce compound 27, compound 27 reacts with carbon tetrabromide to produce compound 28, compound 28 reacts with trimethylsilylacetylene via Sonogashira coupling to produce compound 29, compound 29 reacts to produce compound 30, compound 30 reacts with R1I via Sonogashira coupling to produce compound 31, compound 31 reacts with potassium hydroxylamine to produce compound 32, and compound 32 is deprotected to produce compounds B4-B6; The reaction formula is as follows: Wherein, the substituent R1 in the reaction formula is the same as the substituent R1 in the corresponding compounds B4-B6; Reagents and conditions in the above reaction formula: a. Di-tert-butyl dicarbonate, triethylamine, dichloromethane, reaction at room temperature; b. Carbon tetrabromide, lithium diisopropylamide, tetrahydrofuran, reaction at -78°C and room temperature; c. Cuprous iodide, triethylamine, bis(triphenylphosphine)palladium dichloride, 1,4-dioxane, reaction at 70°C; d. Tetrabutylammonium fluoride; tetrahydrofuran; reaction at 70°C; e. R1I, cuprous iodide, triethylamine, bis(triphenylphosphine)palladium dichloride, 1,4-dioxane, reaction at 70°C; f. Potassium hydroxylamine, methanol, room temperature reaction; g. Trifluoroacetic acid, dichloromethane, reaction at room temperature; (8) Compound 23 reacts with R2I to generate compound 33, compound 33 reacts with elemental iodine to generate compound 34, compound 34 reacts with trimethylsilylacetylene via Sonogashira coupling to generate compound 35, compound 35 reacts to generate compound 36, compound 36 reacts with R1I via Sonogashira coupling to generate compound 37, and compound 37 reacts with potassium hydroxylamine to obtain compound B7; The reaction formula is as follows: Wherein, the substituents R1 and R2 in the reaction formula are the same as the substituents R1 and R2 in the corresponding compound B7; Reagents and conditions in the above reaction formula: a. R2I, sodium hydride, tetrahydrofuran, room temperature reaction; b. Iodine, n-butyl lithium, tetrahydrofuran, -78 ° C and room temperature reaction; c. Cuprous iodide, triethylamine, bis(triphenylphosphine)palladium dichloride, 1,4-dioxane, reaction at 70°C; d. Tetrabutylammonium fluoride; tetrahydrofuran; reaction at 70°C; e. R1I, cuprous iodide, triethylamine, bis(triphenylphosphine)palladium dichloride, 1,4-dioxane, reaction at 70°C; f. Potassium hydroxylamine, methanol, reaction at room temperature.

4. Use of the HDAC11 isoform selective inhibitor according to any one of claims 1 to 2 in the preparation of a medicament for preventing or treating a disease associated with abnormal HDAC11 expression or activity.

5. The use according to claim 4, characterized in that The disease associated with abnormal HDAC11 expression or activity is cancer, autoimmune disease or metabolic disease.

6. The use according to claim 5, wherein the cancer is liver cancer, myeloproliferative neoplasms, multiple myeloma, Hodgkin's lymphoma, non-small cell lung cancer, glioblastoma, pituitary tumor, prostate cancer, ovarian cancer or acute lymphoblastic leukemia.

7. The use according to claim 5, wherein the autoimmune disease is inflammation, psoriasis, rheumatoid arthritis, rheumatoid arthritis or systemic lupus erythematosus.

8. The use according to claim 5, wherein the metabolic disease is obesity or diabetes.

9. A pharmaceutical composition for preventing or treating cancer, autoimmune disease or metabolic disease, comprising the HDAC11 subtype selective inhibitor according to any one of claims 1 to 2 or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers or excipients.

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