Histone deacetylase inhibitor and application thereof

Abstract

The invention discloses a compound as shown in a formula I and pharmaceutically acceptable salt, isomer, raceme, precursor or solvate thereof. The compound disclosed by the invention can be used for effectively inhibiting histone deacetylase (HDAC), so that the compound can be used as a high-efficiency and low-toxicity medicine for treating diseases related to abnormal regulation of the histone deacetylase. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the field of medicine. Specifically, the present invention relates to a benzylamine compound represented by formula (I) or a pharmaceutically acceptable salt, stereoisomer or prodrug thereof, as well as a pharmaceutical composition containing the compound and the use of the compound or the pharmaceutical composition in preparing a drug. Background Art

[0002] Histone deacetylases (HDACs) are a group of enzymes that regulate a series of biological effects at the cellular chromatin level, including chromatin reorganization, transcriptional activation or inhibition, cell cycle, cell differentiation and apoptosis, by inducing histone deacetylation. They are especially related to the regulation of gene transcription expression after cell activation. Chromatin histone acetylation and deacetylation are one of the key links in regulating gene expression, and abnormal gene expression is the molecular biological basis for the occurrence of tumors and some genetic and metabolic diseases. The degree of histone acetylation is coordinated and controlled by histone acetylases (HATs) and histone deacetylases. The occurrence of tumors is closely related to the imbalance of acetylation and deacetylation of lysine residues at the N-terminus of nucleosome core histones. In vivo, its dynamic balance is maintained by HATs and HDACs. In the transcriptionally active region of chromatin genes, the acetylation activity of its core histones increases; on the contrary, in the transcriptionally inactive region of genes, its acetylation degree is low.

[0003] In humans, there are 18 HDACs, which are divided into two families based on their catalytic mechanism. Eleven of these HDACs are zinc-dependent metalloenzymes, called HDAC 1-11, which use water as a nucleophile to hydrolyze amide bonds. The remaining seven sirtuins 1-7 use NAD+ as a cofactor and transfer the acyl group to the C2 position of the ribose. Human histone deacetylases are further subdivided into four categories based on their sequence homology and cellular localization: Class I = HDAC1, HDAC2, HDAC3, and HDAC8; Class IIa = HDAC4, HDAC5, HDAC7, and HDAC 9; Class IIb = HDAC6 and HDAC10; Class III = sirtuins 1-7; Class IV = HDAC11. Among them, I, II, and IV are classic families and are Zn2+-dependent HDACs. The vast majority of HDAC inhibitors currently under clinical study can inhibit multiple subtypes of HDACs, which often belong to the Zn2+-dependent HDAC family.

[0004] Histone deacetylase inhibitors (HDAC inhibitor, HDACIs) are a class of compounds that inhibit the activity of HDACs. Experiments have shown that HDAC inhibitors increase the acetylation level of chromatin histones, thereby activating the expression of specific genes such as p21, p53, etc., which in turn leads to terminal differentiation of cells or apoptosis of cancer cells. Therefore, HDAC has become one of the hottest targets in the field of tumor chemotherapy drug research and development.

[0005] Since the therapeutic effects of the currently available histone deacetylase inhibitors are uncertain and their toxic side effects are significant, there is an urgent need in the art for a new low-toxic and highly effective histone deacetylase inhibitor to overcome the existing drug tolerance and improve the clinical effect. Summary of the invention

[0006] The object of the present invention is to provide a histone deacetylase inhibitor and a pharmaceutical composition comprising the histone deacetylase inhibitor. The histone deacetylase inhibitor of the present invention should not only have significant histone deacetylase inhibitory activity, but also have low toxicity, so as to have good safety.

[0007] In a first aspect, the present invention provides a compound of formula I, or a pharmaceutically acceptable salt, isomer, racemate, precursor or solvate thereof:

[0008]

[0009] In the formula,

[0010] R 1 is selected from the group consisting of: hydrogen, substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C3-C6 cycloalkenyl, substituted or unsubstituted C3-C8 lactone, substituted or unsubstituted C1-C10 amide, substituted or unsubstituted C1-C10 amide, substituted or unsubstituted C5-C10 aryl, substituted or unsubstituted C3-C8 heterocyclyl, substituted or unsubstituted C5-C10 aromatic heterocyclyl having 1 to 3 heteroatoms independently selected from N, O or S;

[0011] R 2 Selected from: hydrogen, halogen, cyano,

[0012] Ring A is selected from: a substituted or unsubstituted C5-8 aryl group, a substituted or unsubstituted C5-C10 heteroaryl group having 1 to 3 heteroatoms independently selected from N, O or S, a substituted or unsubstituted C5-C10 heterocyclyl group having 1 to 3 heteroatoms independently selected from N, O or S, a substituted or unsubstituted C5-8 aryl group, or a C5-8 heteroaryl group having 1 or 2 heteroatoms independently selected from N, O or S and a substituted or unsubstituted C5-8 heterocyclyl group having 1 or 2 heteroatoms independently selected from N, O or S.

[0013] In a specific embodiment, R 1 Selected from: hydrogen, substituted or unsubstituted C1-C6 alkyl;

[0014] R 2 Selected from:

[0015] Ring A is selected from: substituted or unsubstituted C5-8 aryl, substituted or unsubstituted C5-8 aryl and substituted or unsubstituted C5-8 heterocyclic group having 1 or 2 heteroatoms independently selected from N, O or S.

[0016] In a preferred embodiment, the "substituted or unsubstituted C5-8 aryl or C5-8 heteroaryl having 1 or 2 heteroatoms independently selected from N, O or S and substituted or unsubstituted C5-8 heterocyclyl having 1 or 2 heteroatoms independently selected from N, O or S" is a substituted or unsubstituted indolyl or isoindolyl.

[0017] In a preferred embodiment, the "substituted or unsubstituted C5-8 aryl or C5-8 heteroaryl having 1 or 2 heteroatoms independently selected from N, O or S and substituted or unsubstituted C5-8 heterocyclyl having 1 or 2 heteroatoms independently selected from N, O or S" is a substituted or unsubstituted indolyl or isoindolyl.

[0018] In a specific embodiment, R 1 Selected from: hydrogen;

[0019] R 2 Selected from:

[0020] Ring A is selected from: substituted or unsubstituted phenyl,

[0021] X is selected from: N, O, S, CH;

[0022] R 3 Selected from: hydrogen, substituted or unsubstituted C1-3 alkyl, substituted or unsubstituted C1-3 alkoxy;

[0023] R 4Selected from: hydrogen, substituted or unsubstituted C5-8 aryl, substituted or unsubstituted C5-8 cycloalkyl;

[0024] R 5 Selected from: hydrogen, substituted or unsubstituted C1-3 alkyl, substituted or unsubstituted benzyl;

[0025] R 6 Selected from: hydrogen, substituted or unsubstituted C1-3 alkyl.

[0026] In a specific embodiment, R 1 Selected from: hydrogen;

[0027] R 2 Selected from:

[0028] Ring A is selected from:

[0029] X is selected from: CH;

[0030] R 3 Selected from: hydrogen, trifluoromethoxy;

[0031] R 4 is selected from: hydrogen, substituted or unsubstituted phenyl;

[0032] R 5 Selected from: hydrogen, substituted or unsubstituted methyl, substituted or unsubstituted benzyl;

[0033] R 6 Selected from: hydrogen, substituted or unsubstituted methyl.

[0034] In a preferred embodiment, the compound represented by formula I does not include the following compounds:

[0035]

[0036] In a specific embodiment, the compound is selected from the following group of compounds, or pharmaceutically acceptable salts, isomers, racemates, precursors or solvates thereof,

[0037]

[0038]

[0039] In a specific embodiment, the compound is selected from the group consisting of:

[0040]

[0041]

[0042] Preferably, the compound is selected from the group consisting of:

[0043]

[0044] In a second aspect, the present invention provides a pharmaceutical composition comprising the compound described in the first aspect, or a pharmaceutically acceptable salt, isomer, racemate, precursor or solvate thereof, and a pharmaceutically acceptable excipient.

[0045] In a preferred embodiment, the pharmaceutical composition is used as a therapeutic drug for diseases associated with abnormal regulation of histone deacetylase or for the treatment of diseases associated with abnormal regulation of histone deacetylase.

[0046] The disease associated with abnormal regulation of histone deacetylase is cancer, neurodegenerative disease, AIDS, Alzheimer's disease, malaria or diabetes.

[0047] In a preferred embodiment, the cancer is selected from the group consisting of lymphoma, non-small cell lung cancer, small cell lung cancer, gastric cancer, pancreatic cancer, breast cancer, prostate cancer, liver cancer, skin cancer, leukemia, and cervical cancer.

[0048] In a third aspect, the present invention provides use of the compound according to the first aspect, or a pharmaceutically acceptable salt, isomer, racemate, precursor or solvate thereof in the preparation of an HDAC inhibitor.

[0049] In a preferred embodiment, the HDAC is HDAC1 and / or HDAC6.

[0050] In a specific embodiment, the HDAC inhibitor is a drug for treating diseases associated with abnormal regulation of histone deacetylase.

[0051] In a specific embodiment, the disease associated with abnormal regulation of histone deacetylase is cancer, neurodegenerative disease, AIDS, Alzheimer's disease, malaria or diabetes.

[0052] In a preferred embodiment, the cancer is selected from the group consisting of lymphoma, non-small cell lung cancer, small cell lung cancer, gastric cancer, pancreatic cancer, breast cancer, prostate cancer, liver cancer, skin cancer, leukemia, and cervical cancer.

[0053] In a fourth aspect, the present invention provides a method for inhibiting HDAC, comprising the step of contacting an effective amount of the compound according to the first aspect, or a pharmaceutically acceptable salt, isomer, racemate, precursor or solvate thereof, or the pharmaceutical composition according to claim 7 with HDAC, thereby inhibiting the activity of HDAC.

[0054] In a preferred embodiment, the HDAC is HDAC1 and / or HDAC6.

[0055] In a preferred embodiment, the method of inhibiting HDAC is performed in vivo, or in vitro.

[0056] In a preferred embodiment, the method of inhibiting HDAC is performed in vitro and for non-therapeutic purposes.

[0057] In a fifth aspect, the present invention provides a method for treating diseases associated with abnormal regulation of histone deacetylase, comprising the step of administering a therapeutically effective amount of the compound described in the first aspect, or a pharmaceutically acceptable salt, isomer, racemate, precursor or solvate thereof, or the pharmaceutical composition described in the second aspect to a subject in need thereof.

[0058] In a preferred embodiment, the disease associated with abnormal regulation of histone deacetylase is cancer, neurodegenerative disease, AIDS, Alzheimer's disease, malaria or diabetes.

[0059] In a preferred embodiment, the cancer is selected from the group consisting of lymphoma, non-small cell lung cancer, small cell lung cancer, gastric cancer, pancreatic cancer, breast cancer, prostate cancer, liver cancer, skin cancer, leukemia, and cervical cancer.

[0060] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. BRIEF DESCRIPTION OF THE DRAWINGS

[0061] Figure 1 The effects of panobinostat, compound 8, and compound 13 on zebrafish heart rate are shown;

[0062] Figure 2 The effects of panobinostat, compound 8, and compound 13 on the body length of zebrafish are shown;

[0063] Figure 3 The effects of panobinostat, compound 8, and compound 13 on the yolk sac percentage of zebrafish are shown;

[0064] Figure 4 The effects of panobinostat, compound 8, and compound 13 on the pericardial percentage of zebrafish are shown; and

[0065] Figure 5 The side of zebrafish observed under bright field at different doses of panobinostat, compound 8 and compound 13 are shown. DETAILED DESCRIPTION

[0066] After extensive and in-depth research, the inventors found that one of the main causes of panobinostat cardiotoxicity is that it is off-target to S1PR2 and S1PR3. Further, the inventors designed and synthesized a series of HDAC inhibitors that have not been reported in the literature, and carried out structural characterization, kinase activity tests, S1PR2 and S1PR3 agonist activity tests, and obtained a batch of compounds with HDAC inhibitory activity. These compounds can inhibit the activity of HDAC1 and HDAC6, while not acting on S1PR2 and S1PR3. Among them, the IC50 values ​​of the inhibitory activity of compounds 8 and 13 on HDAC1 and HDAC6 reached the nM level, and the agonist activity on S1PR2 and S1PR3 was very weak, and almost no toxicity was produced. The present invention was completed on this basis.

[0067] Definition of terms

[0068] Relevant terms such as "alkyl", "aryl", "heteroaryl", "halogen", "acyl" and the like used herein are not significantly different from the general meanings of the terms in the art.

[0069] For example, the term "alkyl" refers to a straight chain or a branched chain, and C1-n alkyl refers to a saturated aliphatic hydrocarbon group of 1-n carbon atoms, including straight chains and branched chains. For example, "C1-12 alkyl" means that the group is an alkyl group, and the number of carbon atoms on the carbon chain of the alkyl group is between 1 and 12. It should be noted that when there is no special restriction on the number of carbon atoms, it only refers to the number of carbon atoms of the alkyl part indicated therein, and does not include the number of carbon atoms on the substituent of the alkyl group.

[0070] Those skilled in the art should know the meanings of the following terms or abbreviations.

[0071] The term "pharmaceutically acceptable salt" refers to salts that are suitable for contact with mammalian tissues, especially human tissues, within the scope of reasonable medical judgment without excessive toxicity, irritation, allergic response, etc. and commensurate with a reasonable benefit / risk ratio. For example, medically acceptable salts of amines, carboxylic acids and other types of compounds are well known in the art.

[0072] The term "isomers" refers to two or more compounds that have the same molecular composition but different structures and properties.

[0073] The term "racemate" refers to an equimolar mixture of an optically active chiral molecule and its enantiomer, which is composed of equal amounts of molecules with opposite optical rotation directions and the same optical rotation ability. The optical activities of the mixture are offset by the interaction between these molecules, so the mixture is optically inactive.

[0074] The term "solvate" refers to a mixture of a compound and a solvent. For example, a crystal is a solvate.

[0075] Histone deacetylase inhibitors

[0076] The term "histone deacetylase inhibitor" used herein refers to a substance that has an inhibitory effect on histone deacetylase, especially a small molecule compound that has an inhibitory effect on histone deacetylase.

[0077] Histone deacetylase inhibitors generally include three parts: zinc ion binding region, linker region and surface recognition region. The direct interaction between the inhibitor and zinc ion is necessary to produce inhibitory activity. HDAC inhibitors mainly include the following four categories: (1) short-chain fatty acids, such as butyric acid, phenylbutyric acid and their salts; (2) hydroxamic acids, which are the most widely studied class of HDAC inhibitors, such as suberoylanilide hydroxamic acid (SAHA) and trichostatin A (TSA); (3) cyclotetrapeptides, which are the most complex class of inhibitors. In the molecules of cycloteptide inhibitors, the amino acid macrocycle serves as the hydrophobic surface recognition region, the alkyl chain serves as the connecting region, and the end of the alkyl chain is connected to a zinc ion binding group, such as trapoxin, HC-toxon, Apicidin, FK228 and Largazole; (4) benzamide compounds, which have lower activity than general hydroxamic acids and cyclopeptides, but have higher selectivity for class I HDAC, such as MS-275, CI-994, etc. As a new generation of targeted anti-tumor drugs, histone deacetylase inhibitors have the advantage of low cytotoxicity compared with traditional anti-tumor drugs.

[0078] Compounds of the present invention

[0079] The present invention provides a series of HDAC inhibitors with novel structures. These HDACy inhibitors can not only inhibit the activities of HDAC1 and HDAC6, but also do not act on S1PR2 and S1PR3, thus having extremely low toxicity.

[0080] In a specific embodiment, the histone deacetylase inhibitor of the present invention is a compound represented by Formula I, or a pharmaceutically acceptable salt, isomer, racemate, precursor or solvate thereof:

[0081]

[0082] In the formula,

[0083] R 1 , R 2 and Ring A is as described above.

[0084] In a preferred embodiment, the compound of the present invention is a compound selected from the group consisting of

[0085]

[0086] More preferably, the compound of the present invention is a compound selected from the group consisting of:

[0087]

[0088] Most preferably, the compound of the present invention is a compound selected from the group consisting of:

[0089]

[0090]

[0091] Pharmaceutical composition

[0092] On the basis of the compound with HDAC inhibitory activity of the present invention, the present invention also provides a drug combination. The pharmaceutical composition comprises the compound of the present invention, or its pharmaceutically acceptable salt, isomer, racemate, precursor or solvate, and a pharmaceutically acceptable excipient. Based on the teachings of the present invention and the common knowledge in the art, those skilled in the art can know that the pharmaceutical composition of the present invention is used as a therapeutic drug for diseases related to abnormal regulation of histone deacetylase or for the treatment of diseases related to abnormal regulation of histone deacetylase.

[0093] use

[0094] Based on the teachings of the present invention, those skilled in the art will appreciate that the compounds of the present invention, or their pharmaceutically acceptable salts, isomers, racemates, precursors or solvates can be used to prepare HDAC inhibitors. The HDAC inhibitors can be drugs for treating diseases associated with abnormal regulation of histone deacetylase, and can also be used in other fields, such as scientific research. Therefore, the HDAC inhibitors of the present invention can have medical uses or non-medical uses.

[0095] In a specific embodiment, the disease associated with abnormal regulation of histone deacetylase is cancer, neurodegenerative disease, AIDS, Alzheimer's disease, malaria or diabetes. In a preferred embodiment, the cancer is selected from: lymphoma, non-small cell lung cancer, small cell lung cancer, gastric cancer, pancreatic cancer, breast cancer, prostate cancer, liver cancer, skin cancer, leukemia, cervical cancer.

[0096] How to use

[0097] Based on the HDAC inhibitor of the present invention, those skilled in the art will be able to know various methods of using the HDAC inhibitor. For example, in the field of scientific research, an effective amount of the compound of the present invention, or a pharmaceutically acceptable salt, isomer, racemate, precursor or solvate thereof, or a pharmaceutical composition can be contacted with HDAC in vitro or in vivo to inhibit HDAC activity. This method of inhibiting HDAC can be performed in vitro and is non-therapeutic.

[0098] The compounds of the present invention, or pharmaceutically acceptable salts, isomers, racemates, precursors or solvates thereof, can also be used in therapeutic methods. In a specific embodiment, a therapeutically effective amount of the compounds of the present invention, or pharmaceutically acceptable salts, isomers, racemates, precursors or solvates thereof, or a pharmaceutical composition can be administered to a subject in need thereof, thereby treating diseases associated with abnormal regulation of histone deacetylase.

[0099] Beneficial technical effects or advantages of the present invention:

[0100] 1. The HDAC inhibitor of the present invention is a compound with novel structure;

[0101] 2. The kinase activity of the HDAC inhibitor of the present invention is maintained at the same level as that of the lead compound panobinostat;

[0102] 3. The toxicity of the HDAC compounds of the present invention is significantly reduced;

[0103] 4. The compounds of the present invention lay a new material foundation for the development of highly effective, low-toxic therapeutic drugs for diseases related to abnormal histone deacetylase regulation, thus having excellent application potential and social value.

[0104] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples that do not specify specific conditions are usually based on conventional conditions or the conditions recommended by the manufacturer. The reagents used in the examples are all commercially available reagents. Unless otherwise stated, percentages and parts are calculated by weight.

[0105] Example

[0106] Materials and Methods

[0107] The synthesis of the compound of the present invention is as follows:

[0108] Example 1

[0109] The synthesis of the compound of the present invention is as follows:

[0110] Synthesis of 4-(((2-(2-methyl-1H-indol-3-yl)ethyl)amino)methyl)benzaldehyde (Compound 1-1b) (Step a)

[0111]

[0112] At 0°C, 2-methyltryptamine (5.0 g, 28.69 mmol) and potassium carbonate (7.9 g, 57.39 mmol) were suspended in 20 mL of DMF, and then 4-chloromethylbenzaldehyde (1.5 g, 9.56 mmol) was added thereto, stirred for 2 h, and the reaction progress was monitored by thin layer chromatography. After the reaction was completed, water was added to the reaction solution to quench the reaction, and then ethyl acetate was used for extraction. The organic phase was washed with water and saturated sodium chloride in sequence, and then dried over anhydrous sodium sulfate, and concentrated under reduced pressure to remove the solvent to obtain a colorless oily liquid. The crude product was then purified by column chromatography (DCM / MeOH: 30 / 1) to obtain 1.4 g of a light yellow solid with a yield of 50%.

[0113] 1 H NMR (500MHz, Methanol-d 4 )δ7.89(s,1H),7.59(d,J=8.1Hz,2H),7.46(d,J=7.6Hz,1H),7.41(d,J=7.6Hz,1H),7.35(d,J=8.1Hz,2H),7.25 (d,J=7.2Hz,1H),7.22(d,J=7.2Hz,1H),3.92(s,2H),3.07(t,J=6.7Hz,2H),2.94(d,J=6.7Hz,2H),2.37(s,3H).

[0114] Synthesis of 2-((4-(((2-(2-methyl-1H-indol-3-yl)ethyl)amino)methyl)benzyl)amino)ethanol (Compound 1) (Step b)

[0115]

[0116] 4-(((2-(2-methyl-1H-indol-3-yl)ethyl)amino)methyl)benzaldehyde (100 mg, 0.34 mmol) was dissolved in 5 mL of methanol, 1 drop of acetic acid was added, and then ethanolamine (42 mg, 0.68 mmol) was added. After stirring at room temperature for 3 minutes, sodium cyanoborohydride (22 mg, 0.34 mmol) was added. TLC was used to monitor the progress of the reaction. After the reaction was complete, the reaction system was distilled under reduced pressure, and water and dichloromethane were added for extraction. The mixture was washed with saturated brine, the organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by PTLC to obtain 20 mg of a white solid with a yield of 17%.

[0117] 1H NMR (600 MHz, DMSO-d 6 )δ10.73(s,1H),7.37(dd,J=7.7,1.2Hz,1H),7.34-7.29(m,4H),7.22-7.20(m,1H),6.98-6.94(m,1H),6.91-6.88(m,1H),4.61(s,2H), 3.83(s,2H),3.75(s,2H),3.49(t,J=5.7Hz,2H),3.17(s,1H),2.86-2.84(m,2H),2.79-2.74(m,2H),2.60(t,J=5.8Hz,2H),2.30(s,3H). 13 C NMR (151 MHz, DMSO-d 6 )δ135.17,132.00,128.30(2C),128.19(4C),119.92,118.07,117.31,110.38,107.44, 59.67,52.11,51.81,50.56,48.94,48.60,23.34,11.27.LC-MS(ESI):m / z:338.20(M+H) + .HRMS(ESI)(m / z):[M+H] + calcdfor C 21 H 28 N 3 O 338.2232; found:338.2233.

[0118] Synthesis of 4-(((2-(2-methyl-1H-indol-3-yl)ethyl)amino)methyl)phenol (Compound 2-2b) (Step a)

[0119]

[0120] At 0°C, 2-methyltryptamine (500 mg, 2.87 mmol) and potassium carbonate (793 mg, 5.74 mmol) were suspended in 20 mL of DMF, and then 4-chloromethylphenol (140 mg, 0.96 mmol) was added thereto, stirred for 2 h, and the reaction progress was monitored by thin layer chromatography. After the reaction was completed, water was added to the reaction solution to quench the reaction, and then ethyl acetate was used for extraction. The organic phase was washed with water and saturated sodium chloride in sequence, and then dried over anhydrous sodium sulfate, and concentrated under reduced pressure to remove the solvent to obtain a colorless oily liquid. The crude product was purified by column chromatography to obtain 100 mg of a light yellow solid with a yield of 37%.

[0121] Synthesis of 3-(4-(((2-(2-methyl-1H-indol-3-yl)ethyl)amino)methyl)phenoxy)propane-1,2-diol (Compound 2) (Step b)

[0122]

[0123] At 0°C, 4-(((2-(2-methyl-1H-indol-3-yl)ethyl)amino)methyl)phenol (100 mg, 0.36 mmol) and cesium carbonate (138 mg, 0.71 mmol) were suspended in 20 mL of DMF, and then 3-chloropropane-1,2-diol (118 mg, 1.07 mmol) was added thereto, stirred for 2 h, and the reaction progress was monitored by thin layer chromatography. After the reaction was completed, water was added to the reaction solution to quench the reaction, and then extracted with dichloromethane. The organic phase was washed with water and saturated sodium chloride in sequence, and then dried over anhydrous sodium sulfate, and concentrated under reduced pressure to remove the solvent to obtain a colorless oily liquid. The crude product was purified by column chromatography to obtain 100 mg of a light yellow solid with a yield of 79%.

[0124] 1 H NMR (600 MHz, DMSO-d 6 )δ10.82(s,1H),7.40(t,J=9.1Hz,3H),7.23(d,J=8.0Hz,1H),7.01-6.90(m,4H),4.97(d,J=5.1Hz,1H),4.69(t,J=5.7Hz,1H) ,4.01-3.96(m,3H),3.87-3.83(m,1H),3.80-3.76(m,1H),3.44(t,J=5.1Hz,2H),3.17(s,1H),2.98-2.87(m,4H),2.32(s,3H). 13 C NMR (151 MHz, DMSO-d 6 )δ158.82,135.20,132.43,130.91(2C),127.91,120.14,118.26,117.20 ,114.43(2C),110.51,69.92,69.67,62.69,50.11,48.61,47.39,11.22.

[0125] Synthesis of 1-(4-(((2-(2-methyl-1H-indol-3-yl)ethyl)amino)methyl)benzyl)pyrrolidine-3-carboxylic acid (Compound 3)

[0126]

[0127] Referring to the synthesis method of compound 1, 84 mg of white solid was obtained with a yield of 63.2%.

[0128] 1 H NMR (600 MHz, DMSO-d 6 )δ10.85(s,1H),7.49(d,J=7.8Hz,2H),7.42(d,J=7.8Hz,1H),7.33(d,J=7.8Hz,2H),7.23(d,J=8.0Hz,1H),6.99-6.96(m,1H),6.93-6.89(m,1 H),4.07(s,2H),3.80(s,2H),3.63-3.54(m,4H),3.03-2.99(m,2H),2. 68(t,J=8.8Hz,2H),2.62-2.58(m,2H),2.32(s,3H),1.97-1.92(m,2H). 13 C NMR (151 MHz, DMSO-d 6 )δ172.04,135.20,132.46,129.56,128.67(2C),127.87,120.13,118.23,117 .22,110.49,105.71,58.70,56.14,53.26,49.97,47.27,27.11,21.13,11.17.

[0129] Synthesis of (S)-1-(4-(((2-(2-methyl-1H-indol-3-yl)ethyl)amino)methyl)benzyl)pyrrolidine-3-carboxylic acid (Compound 4)

[0130]

[0131] The synthetic method of reference compound 1 was used to obtain 90 mg of a white solid with a yield of 67.7%.

[0132] 1 H NMR (600 MHz, DMSO-d 6)δ10.70(s,1H),7.35(d,J=7.8Hz,1H),7.27(d,J=7.8Hz,2H),7.23-7.18(m,3H),6.9 7-6.92(m,1H),6.91-6.85(m,1H),3.76(s,2H),3.56-3.48(m,2H),3.17(s,1H),2.90 -2.83(m,1H),2.81(t,J=7.6Hz,2H),2.73-2.68(m,2H),2.67(t,J=8.8Hz,1H),2.56( dd,J=9.2,6.5Hz,1H),2.48(d,J=7.7Hz,1H),2.42(q,J=7.8Hz,1H),2.29(s,3H),1.96 -1.90(m,2H). 13 C NMR (151 MHz, DMSO-d 6 )δ176.23,138.22,137.54,135.16,131.86,128.33(2C),128.28,127.99(2C),119.84,118. 00,117.32,110.33,107.87,58.99,56.48,53.34,52.14,49.22,45.58,27.27,23.86,11.25.

[0133] Synthesis of 1-(4-(((2-(2-methyl-1H-indol-3-yl)ethyl)amino)methyl)benzyl)azetidine-3-carboxylic acid (Compound 5)

[0134]

[0135] The synthetic method of reference compound 1 was used to obtain 92 mg of a white solid with a yield of 71.1%.

[0136] 1 H NMR (600 MHz, Methanol-d 4 )δ7.51(d,J=8.0Hz,2H),7.49(d,J=8.5Hz,2H),7.42-7.41(m,1H),7.25(dt,J=8.1,0.9Hz,1H),7.06-7.00(m,1H),6.98-6.95(m,1H),4.21(s ,2H),4.19(s,2H),4.12-4.08(m,1H),4.01(t,J=9.4Hz,2H),3.94(dd,J=9.7,7.5Hz,2H),3.22-3.15(m,2H),3.11-3.06(m,2H),2.39(s,3H).13 C NMR (151MHz, Methanol-d 4 )δ177.76,137.23,134.61,134.45,133.88,131.56(2C),131.52(2C),131.07,130.96,129 .30,125.10,121.80,119.87,117.98,111.61,105.97,60.01,58.22,36.40,22.44,11.30.

[0137] Synthesis of 2-(2-methyl-6-(trifluoromethoxy)-1H-indol-3-yl)ethyl-1-amine (Compound 6-6b) (Step a)

[0138]

[0139] 3-Trifluoromethoxyphenylhydrazine hydrochloride (1000mg, 4.37mmol) was placed in a 100mL single-necked flask, and a mixed solution of ethanol and water (28mL, 14:1) was added, and stirred and refluxed at 120°C for 30 minutes. Then 5-chloro-2-pentanone (527mg, 4.37mmol) was dissolved in 3mL of ethanol and added, and stirred at 120°C for about 16h. The reaction progress was monitored by thin layer chromatography. After the reaction was completed, pH=10 sodium hydroxide solution was added to adjust the reaction solution to alkalinity, and the reaction solution was extracted with EA 3 times. The organic phase was washed once with water and saturated brine, and the aqueous phase was stripped with EA, and the organic phases were combined. Purification by silica gel column chromatography (DCM: MeOH=15:1) gave 701mg of brown oily liquid.

[0140] LC-MS / ESI[M+H] + 259.10.

[0141] Synthesis of (E)-3-(4-(((2-(2-methyl-6-(trifluoromethoxy)-1H-indol-3-yl)ethyl)amino)methyl)phenyl)-N-(tetrahydro-2H-pyran-2-yl)oxy)acrylamide (Compound 6-6c) (Step b)

[0142]

[0143] Take 2-(2-methyl-6-(trifluoromethoxy)-1H-indol-3-yl)ethyl-1-amine (701mg, 2.71mmol) in a 100mL single-necked flask, add 10mL DCM and 3mL methanol to dissolve it, add (E)-3-(4-formyl-2-methylphenyl)-N-((tetrahydro-2H-pyran-2-yl)oxy)acrylamide (623mg, 2.26mmol) and stir at room temperature for 30min, then add sodium triacetyl borohydride (156mg, 2.49mmol), continue stirring, and monitor the reaction progress by thin layer chromatography. After the reaction is complete, the reaction solution is spin-dried, the reaction solution is extracted with DCM 3 times, the organic phase is washed once with water and saturated brine, the aqueous phase is back-extracted with DCM, and the organic phases are combined. Purification by silica gel column chromatography (DCM: MeOH = 15: 1) to obtain 314mg of a yellow oily liquid.

[0144] LC-MS / ESI[M+H] + 518.20.

[0145] Synthesis of (E)-N-hydroxy-3-(4-(((2-(2-methyl-6-(trifluoromethoxy)-1H-indol-3-yl)ethyl)amino)methyl)phenyl)acrylamide (Compound 6) (Step c)

[0146]

[0147] (E)-3-(4-(((2-(2-methyl-6-(trifluoromethoxy)-1H-indol-3-yl)ethyl)amino)methyl)phenyl)-N-(tetrahydro-2H-pyran-2-yl)oxy)acrylamide (314 mg, 0.61 mmol) was placed in a 100 mL single-necked flask, 5 mL of DCM was added to dissolve it, and dioxane hydrochloride (1.51 mL, 6.01 mmol) was added. The mixture was stirred at room temperature to precipitate a solid. The reaction progress was monitored by thin layer chromatography. After the reaction was complete, the solid was filtered, washed with DCM, and filtered again to collect the solid. 120 mg of a yellow solid was obtained, with a yield of 45.6%.

[0148] 1 H NMR (500 MHz, DMSO-d 6)δ11.26(s,1H),9.72(s,2H),7.64(d,J=8.0Hz,2H),7.59(d,J=7.7Hz,2H), 7.56(d,J=8.5Hz,1H),7.47(d,J=5.9Hz,1H),7.41(d,J=29.1Hz,1H),7.28- 7.19(m,1H),6.91(dd,J=8.5,2.1Hz,1H),6.58(d,J=15.9Hz,1H),4.16(t,J =6.0Hz,2H),3.12(dd,J=10.0,6.4Hz,2H),3.01-2.95(m,2H),2.34(s,3H). 13 C NMR (126 MHz, DMSO-d 6 )δ172.06,152.49,147.02,144.84,144.36,144.22,142.84,140.15(2C),137.11(2C),136.41,129.98 (q,J=252.0Hz),129.56,127.69,121.49,115.22,113.03,58.93,56.41,29.88,20.78.LC-MS / ESI[M+H] + 434.10.

[0149] Synthesis of 3-((4-(((2-(2-methyl-1H-indol-3-yl)ethyl)amino)methyl)benzyl)amino)propionic acid (Compound 7)

[0150]

[0151] The synthetic method of reference compound 1 was used to obtain 11 mg of a white solid with a yield of 8.9%.

[0152] 1 H NMR (600 MHz, Methanol-d 4)δ7.53(s,2H),7.49(d,J=8.6Hz,1H),7.46(t,J=4.0Hz,1H),7.41(d,J=7.8Hz,1H),7 .34(dd,J=8.1,4.1Hz,1H),7.25(dd,J=8.1,5.1Hz,1H),7.03-7.00(m,J=7.1,3.5Hz, 1H),6.98-6.94(m,1H),5.49(s,1H),4.19(d,J=8.3Hz,1H),4.16-4.08(m,2H),3.35( s,1H),3.15-3.12(m,2H),3.07(t,J=6.4Hz,4H),2.46(t,J=6.4Hz,3H),2.38(s,3H). 13 C NMR (151MHz, Methanol-d 4 )δ178.87,134.36,133.83,131.57(2C),131.42,131.36(2C),131.21,129.34,12 1.73,119.81,118.04,111.57,38.04,34.29,32.99,23.69,23.23,22.98,11.31.

[0153] Synthesis of (E)-3-(4-formylphenyl)-N-((tetrahydro-2H-pyran-2-yl)oxy)acrylamide (Compound 8-8b) (Step a)

[0154]

[0155] Take (E)-3-(4-formylphenyl) methyl acrylate (500mg, 2.84mmol) in a 100mL single-necked flask, add 10mL DMF to dissolve it, then add HATU (1295mg, 3.40mmol), DIPEA (1.03mL, 5.96mmol) and stir evenly at room temperature, then add O-(tetrahydro-2H-pyran-2-yl) hydroxylamine (333mg, 2.83mmol), continue stirring, and the reaction is complete in about 30 minutes. The reaction solution is extracted with ethyl acetate 3 times, the organic phase is washed once with water and saturated brine, the aqueous phase is back-extracted with ethyl acetate, and the organic phases are combined. Purification by silica gel column chromatography (PE:EA=5:1) gives 472mg of a white solid with a yield of 60.6%.

[0156] 1 H NMR (500 MHz, DMSO-d 6)δ11.36(s,1H),10.02(s,1H),7.97-7.92(m,2H),7.80(d,J=7.9Hz,2H),7.57(d,J=15.8Hz,1H),6.66(d,J=1 5.9Hz,1H),4.93(s,1H),3.98(d,J=10.5Hz,1H),3.57-3.51(m,1H),1.70(s,3H),1.54(s,3H).LC-MS / ESI[MH] - 274.15.

[0157] Synthesis of tert-butyl (2-(2-methyl-1H-indol-3-yl)ethyl)carbamate (Compound 8-8c) (Step b)

[0158]

[0159] Take 1H-2-methyl-3-aminoethyl indole (500mg, 2.87mmol) in a 100mL single-necked flask, add 5mL methanol to dissolve it, add di-tert-butyl dicarbonate (637mg, 2.87mmol) and triethylamine (0.45mL, 3.15mmol), stir at room temperature, and monitor the reaction process by thin layer chromatography. After the reaction is complete, extract the reaction solution with ethyl acetate 3 times, wash the organic phase with water and saturated brine once each, extract the aqueous phase with ethyl acetate, and combine the organic phases. Purify by silica gel column chromatography (DCM: MeOH = 100: 1) to obtain 817mg of yellow oily liquid, with a yield of 96.9%.

[0160] 1 H NMR(500MHz,Chloroform-d)δ7.94(s,1H),7.49(d,J=7.7Hz,1H),7.14-7.11(m,1H),7.09-7.06 (m,1H),3.37(q,J=6.7Hz,2H),2.89(t,J=6.7Hz,2H),2.37(s,3H),1.44(s,9H).LC-MS / ESI[M+H] + 275.15.

[0161] Synthesis of tert-butyl (2-(1-phenyl-2-methyl-1H-indol-3-yl)ethyl)carbamate (Compound 8-8d) (Step c)

[0162]

[0163] Take tert-butyl (2-(2-methyl-1H-indol-3-yl) ethyl) carbamate (817mg, 2.98mmol) in a 100mL single-necked flask, add 12mL DMF to dissolve it, add sodium hydride (79mg, 3.27mmol) and stir at room temperature for 10min, then add benzyl bromide (0.4mL, 3.27mmol), continue stirring, and monitor the reaction progress by thin layer chromatography. After the reaction is complete, add water to quench, extract the reaction solution with ethyl acetate 3 times, wash the organic phase with water and saturated brine once each, extract the aqueous phase with ethyl acetate, and combine the organic phases. Purify by silica gel column chromatography (DCM: MeOH = 100: 1) to obtain 810mg of a yellow oily liquid with a yield of 74.5%.

[0164] LC-MS / ESI[M+H] + 365.20. 1 H NMR (400 MHz, DMSO-d 6 )δ7.50-7.45(m,1H),7.33-7.30(m,1H),7.29-7.25(m,2H),7.23-7.20(m,1H),7.03-6.98(m,2H),6.98-6.96 (m,2H),6.84(t,J=5.7Hz,1H),5.38(s,2H),3.12-3.02(m,2H),2.82-2.78(m,2H),2.28(s,3H),1.36(s,9H).

[0165] Synthesis of 2-(1-phenyl-2-methyl-1H-indol-3-yl)ethyl-1-amine (Compound 8-8f) (Step d)

[0166]

[0167] Take tert-butyl (2- (1-phenyl-2-methyl-1H-indol-3-yl) ethyl) carbamate (810mg, 2.22mmol) in a 100mL single-necked flask, add 8mL of dioxane to dissolve it, add 2mL of 12M hydrochloric acid, reflux at 110°C, and monitor the reaction progress by thin layer chromatography. After the reaction is complete, the reaction solution is spin-dried and purified by silica gel column chromatography (DCM: MeOH = 15: 1) to obtain 590mg of brown solid, with a yield of 100.4%.

[0168] LC-MS / ESI[M+H] + 265.20.

[0169] Synthesis of (E)-3-(4-(((2-(1-phenyl-2-methyl-1H-indol-3-yl)ethyl)amino)methyl)phenyl)-N-((tetrahydro-2H-pyran-2-yl)oxy)acrylamide (Compound 8-8e) (Step f)

[0170]

[0171] Take 2-(1-phenyl-2-methyl-1H-indol-3-yl)ethyl-1-amine (409mg, 1.55mmol) in a 100mL single-necked flask, add 10mL dichloromethane and 3mL methanol to dissolve it, add (E)-3-(4-formylphenyl)-N-((tetrahydro-2H-pyran-2-yl)oxy)acrylamide (426mg, 1.55mmol) and stir at room temperature for 30min, then add sodium cyanoborohydride (94mg, 1.48mmol), continue stirring, and monitor the reaction process by thin layer chromatography. After the reaction is complete, the reaction solution is spin-dried, the reaction solution is extracted with dichloromethane 3 times, the organic phase is washed with water and saturated brine once each, the dichloromethane back-extracts the aqueous phase, and the organic phase is combined. Purification by silica gel column chromatography (DCM: MeOH = 50: 1) gives 199mg of brown solid, with a yield of 24.5%.

[0172] LC-MS / ESI[M+H] + 524.30.

[0173] Synthesis of (E)-3-(4-(((2-(1-phenyl-2-methyl-1H-indol-3-yl)ethyl)amino)methyl)phenyl)-N-hydroxyacrylamide (Compound 8) (Step g)

[0174]

[0175] (E)-3-(4-(((2-(1-phenyl-2-methyl-1H-indol-3-yl)ethyl)amino)methyl)phenyl)-N-((tetrahydro-2H-pyran-2-yl)oxy)acrylamide (199 mg, 0.38 mmol) was placed in a 100 mL single-necked flask, 5 mL of dichloromethane was added to dissolve it, 0.85 mL of dioxane hydrochloride was added, and the mixture was stirred at room temperature to precipitate a solid. The reaction progress was monitored by thin layer chromatography. After the reaction was complete, the mixture was filtered, the solid was washed with dichloromethane, and the solid was filtered again to collect the solid. 63 mg of a yellow solid was obtained, with a yield of 37.7%.

[0176] 1 H NMR (500MHz, Methanol-d 4)δ7.49(d,J=7.1Hz,1H),7.46(s,1H),7.43(s,1H),7.42(s,1H),7.23-7.17(m,5H),7.04(t,J=6.9Hz,1H),6.98(t,J=7.4Hz,1 H), 6.90 (s, 1H), 6.89 (s, 1H), 6.44 (d, J = 15.8Hz, 1H), 5.32 (s, 2H), 2.98 (t, J = 7.3Hz, 2H), 2.83 (t, J = 7.3Hz, 2H), 2.25 (s, 3H). 13 C NMR (126MHz, Methanol-d 4 )δ166.20,142.13,140.01,139.90,138.11,135.43,134.56,129.88(2C),129.66(2C),129.24,128.74(2C),128.14 ,127.06(2C),121.92,120.03,119.02,118.82,110.15,109.82,53.84,50.25,47.17,25.18,10.27.LC-MS / ESI[M+H] + 440.20.HRMS(ESI)m / z:calcd for C 28 H 29 N 3 O 2 [M+H] + :439.2260; found:440.2343.

[0177] The synthetic route of target compounds 9-11 is similar to that of compound 8

[0178] Synthesis of (E)-3-(4-(((2-(1,2-dimethyl-1H-indol-3-yl)ethyl)amino)methyl)phenyl)-N-hydroxyacrylamide (Compound 9)

[0179]

[0180] 177 mg of green solid, yield 93.4%.

[0181] 1 H NMR (500MHz, Methanol-d 4)δ7.65(s,1H),7.63(s,1H),7.59(d,J=15.8Hz,1H),7.55(s,1H),7.53(s,1H),7.48(d,J=7.8Hz,1H),7.29(d,J=8.1Hz,1H),7.11(t,J=7.1 Hz,1H),7.01(t,J=7.0Hz,1H),6.56(d,J=15.8Hz,1H),4.25(s,2H),3.66(s,3H),3.20(d,J=5.5Hz,2H),3.18(d,J=5.4Hz,2H),2.40(s,3H). 13 C NMR (126MHz, Methanol-d 4 )δ165.80,141.11,138.29,137.25,135.57,134.16,131.64(2C),129.55,128.45,121.96,120 .10,119.06,118.21(2C),109.90,105.73,68.12,51.78,29.75,22.44,10.19.LC-MS / ESI[M+H] + 364.20.HRMS(ESI)m / z:calcd for C 22 H 25 N 3 O 2 [M+H] + :363.1947; found:364.2023.

[0182] Synthesis of (E)-3-(4-(((2-([1,1'-biphenyl]-4-yl)ethyl)amino)methyl)phenyl)-N-hydroxyacrylamide (Compound 10)

[0183]

[0184] 311 mg of white solid, yield 99.0%.

[0185] 1 H NMR (500 MHz, DMSO-d 6 )δ10.90(s,1H),9.61(s,2H),7.67-7.60(m,8H),7.51-7.42(m,3H),7.39-7.31(m,3H), 6.56(d,J=15.8Hz,1H),4.20(t,J=5.2Hz,2H),3.16(d,J=3.7Hz,2H),3.11-3.04(m,2H). 13C NMR (126 MHz, DMSO-d 6 )δ162.53,139.81,138.68,137.52,136.50,135.40,133.11,130.66(2C),129.23(2C),128.94 (2C),127.62,127.39,126.94(2C),126.55(2C),120.07,49.54,47.42,31.02.LC-MS / ESI[M+H] + 373.20.HRMS(ESI)m / z:calcd for C 24 H 24 N 2 O 2 [M+H] + :372.1838; found:373.1915.

[0186] Synthesis of (E)-N-hydroxy-3-(4-((phenylethylamino)methyl)phenyl)acrylamide (Compound 11)

[0187]

[0188] 361 mg of green solid, yield 92.3%.

[0189] 1 H NMR (500 MHz, DMSO-d 6 )δ9.64(s,2H),7.61(s,4H),7.46(d,J=15.8Hz,1H),7.32(t,J=7.6Hz,2H),7.25(t,J=7.3H z,3H),6.56(d,J=15.9Hz,1H),4.17(t,J=5.8Hz,2H),3.16-3.07(m,2H),3.06-3.01(m,2H). 13 C NMR (126 MHz, DMSO-d 6 )δ162.47,137.45,137.30,135.39,133.12,130.65(2C),128.67(2C),128. 59(2C),127.61(2C),126.76,120.08,49.49,47.50,31.39.LC-MS / ESI[M+H] + 297.20.HRMS(ESI)m / z:calcd for C 18 H 20 N 2 O 2 [M+H] +:296.1525; found:297.1599.

[0190] Synthesis of 2-(4-(cyclopent-1-en-1-yl)phenyl)ethyl-1-amine (Compound 12-12b) (Step a)

[0191]

[0192] 4-Bromophenethylamine (500 mg, 2.50 mmol), cyclopenten-1-ylboronic acid (336 mg, 2.50 mmol), Pd(dppf)Cl 2 (183mg, 0.25mmol) and cesium carbonate (1628mg, 5.00mmol) were placed in a 50mL Schlenk tube, and then 20mL of a mixture of DMF and water (DMF:H2O=4:1) was added, and the mixture was heated to 102°C and refluxed with stirring for about 3h under nitrogen protection. After the reaction was completed, diatomaceous earth was filtered and the filtrate was collected. The reaction solution was extracted with ethyl acetate 3 times, the organic phase was washed once with water and saturated brine, the aqueous phase was stripped with ethyl acetate, and the organic phases were combined. Purification by silica gel column chromatography (PE:EA=15:1) gave 199mg of a white solid with a yield of 42.5%.

[0193] Synthesis of 2-(4-cyclopentylphenyl)ethyl-1-amine (Compound 12-12c) (Step b)

[0194]

[0195] 2-(4-(cyclopent-1-ene-1-yl)phenyl)ethyl-1-amine (199mg, 1.06mmol) and palladium carbon (40mg, 0.37mmol) were placed in a 100mL single-necked flask, and a mixed solution of methanol and ethanol (10mL) was added. The mixture was reacted at room temperature for about 5 hours under the pressure of 8 nitrogen balloons. After the reaction was completed, diatomaceous earth was filtered and the filtrate was collected. The reaction solution was extracted with ethyl acetate 3 times, and the organic phase was washed once with water and saturated brine. The aqueous phase was back-extracted with ethyl acetate, and the organic phases were combined. Purification by silica gel column chromatography (DCM:MeOH=30:1) gave 200mg of brown solid with a yield of 99.4%.

[0196] Synthesis of (E)-3-(4-(((4-cyclopentylphenethyl)amino)methyl)phenyl)-N-((tetrahydro-2H-pyran-2-yl)oxy)acrylamide (Compound 12-12d) (Step c)

[0197]

[0198] Take 2-(4-cyclopentylphenyl)ethyl-1-amine (200mg, 1.05mmol) in a 100mL single-necked flask, add 10mL dichloromethane and 3mL methanol to dissolve it, add (E)-3-(4-formylphenyl)-N-((tetrahydro-2H-pyran-2-yl)oxy)acrylamide (291mg, 1.05mmol) and stir at room temperature for 30min, then add sodium cyanoborohydride (66mg, 1.05mmol), continue stirring, and monitor the reaction progress by thin layer chromatography. After the reaction is complete, the reaction solution is spin-dried, the reaction solution is extracted with dichloromethane 3 times, the organic phase is washed with water and saturated brine once each, the aqueous phase is stripped with dichloromethane, and the organic phase is combined. Purification by silica gel column chromatography (DCM: MeOH = 30: 1) gives 291mg of brown oil, with a yield of 61.5%.

[0199] Synthesis of (E)-3-(4-(((4-cyclopentylphenyl)amino)methyl)phenyl)-N-hydroxyacrylamide (Compound 12) (Step d)

[0200]

[0201] (E)-3-(4-(((4-cyclopentylphenethyl)amino)methyl)phenyl)-N-((tetrahydro-2H-pyran-2-yl)oxy)acrylamide (291 mg, 0.65 mmol) was placed in a 100 mL single-necked flask, 5 mL of dichloromethane was added to dissolve it, 1.46 mL of dioxane hydrochloride was added, and the mixture was stirred at room temperature to precipitate a solid. The reaction progress was monitored by thin layer chromatography. After the reaction was complete, the mixture was filtered, the solid was washed with DCM, and the solid was filtered again to collect the solid. 190 mg of brown solid was obtained, with a yield of 80.1%.

[0202] 1 H NMR (500 MHz, DMSO-d 6 )δ9.72(s,2H),7.63(d,J=8.2Hz,2H),7.59(d,J=8.0Hz,1H),7.48(d,J=12.1Hz,1H),7 .41(d,J=22.7Hz,1H),7.29(s,1H),7.18(d,J=7.9Hz,2H),7.12(d,J=7.9Hz,2H),6.58( d,J=15.9Hz,1H),4.15(t,J=5.9Hz,2H),3.09-3.02(m,2H),3.02-2.95(m,2H),2.94-2. 88(m,1H),2.00-1.92(m,2H),1.78-1.67(m,2H),1.66-1.57(m,2H),1.53-1.42(m,2H). 13 C NMR (126 MHz, DMSO-d6 )δ162.53,144.39,137.48,135.39,134.61,133.16,130.73(2C),128.48(2C),127.61(2C) ,127.25(2C),120.11,48.61,47.61,44.99,34.27(2C),31.01,25.04(2C).LC-MS / ESI[M+H] + 365.20.HRMS(ESI)m / z:calcd forC 23 H 28 N 2 O 2 [M+H] + :364.2151; found:365.2228.

[0203] Synthesis of 2-(7-bromo-2-methyl-1H-indol-3-yl)ethyl-1-amine (Compound 13-13b) (Step a)

[0204]

[0205] 2-Bromophenylhydrazine hydrochloride (1000mg, 4.47mmol) was placed in a 100mL single-necked flask, and a mixed solution of ethanol and water (28mL, 14:1) was added, and stirred and refluxed at 120°C for 30 minutes. Then 5-chloro-2-pentanone (539mg, 4.47mmol) was dissolved in 3mL of ethanol and added, and stirred at 120°C for about 16h. The reaction progress was monitored by thin layer chromatography. After the reaction was completed, pH=10 sodium hydroxide solution was added to adjust the reaction solution to alkalinity, and the reaction solution was extracted with EA 3 times. The organic phase was washed once with water and saturated brine, and the aqueous phase was stripped with EA, and the organic phases were combined. Purification by silica gel column chromatography (DCM: MeOH = 15:1) gave 1081mg of brown liquid with a yield of 96.9%.

[0206] LC-MS / ESI[M+H] + 253.05. 1 H NMR (500MHz, Methanol-d 4 )δ7.43-7.40(m,1H),7.17-7.15(m,1H),6.88(t,J=7.7Hz,1H),2.91-2.88(m,2H),2.87-2.84(m,2H),2.41(s,3H).

[0207] Synthesis of tert-butyl (2-(7-bromo-2-methyl-1H-indol-3-yl)ethyl)carbamate (Compound 13-13c) (Step b)

[0208]

[0209] Take 2-(7-bromo-2-methyl-1H-indol-3-yl)ethyl-1-amine (1081mg, 4.27mmol) in a 100mL single-necked flask, add 5mL DCM to dissolve it, add di-tert-butyl dicarbonate (932mg, 4.27mmol) and triethylamine (0.60mL, 4.7mmol), stir at room temperature, and monitor the reaction progress by thin layer chromatography. After the reaction is complete, extract the reaction solution with EA 3 times, wash the organic phase with water and saturated brine once each, extract the aqueous phase with EA, and combine the organic phases. Purify by silica gel column chromatography (DCM: MeOH = 100: 1) to obtain 474mg of brown solid, with a yield of 32.7%.

[0210] 1 H NMR (500MHz, Methanol-d 4 )δ7.45(d,J=7.9Hz,1H),7.17(d,J=7.6Hz,1H),6.88(t,J=7.7Hz,1H),3.23(t, J=7.2Hz,2H),2.85(t,J=7.3Hz,2H),2.42(s,3H),1.43(s,9H).LC-MS / ESI[M+H] + 297.05.

[0211] Synthesis of tert-butyl (2-(2-methyl-7-phenyl-1H-indol-3-yl)ethyl)carbamate (Compound 13-13d) (Step c)

[0212]

[0213] Tert-butyl (2-(7-bromo-2-methyl-1H-indol-3-yl)ethyl)carbamate (474 ​​mg, 1.40 mmol), phenylboronic acid (221 mg, 1.81 mmol), Pd(dppf)Cl 2 (82 mg, 0.11 mmol) and sodium carbonate (296 mg, 2.80 mmol) were placed in a 25 mL Schlenk tube, and then 15 mL of a mixture of DMF and water (DMF:H 2 O=4:1), heated to 102℃ and refluxed with stirring for about 3h under nitrogen protection. After the reaction is completed, filter with diatomaceous earth and collect the filtrate. Extract the reaction solution 3 times with EA, wash the organic phase with water and saturated brine once each, extract the aqueous phase with EA, and combine the organic phases. Purify by silica gel column chromatography (PE:EA=15:1) to obtain 411mg of white solid, with a yield of 83.9%. LC-MS / ESI[M+H] + 295.20.

[0214] Synthesis of 2-(2-methyl-7-phenyl-1H-indol-3-yl)ethyl-1-amine (Compound 13-13e) (Step d)

[0215]

[0216] Take tert-butyl (2-(2-methyl-7-phenyl-1H-indol-3-yl) ethyl) carbamate (411 mg, 1.27 mmol) in a 100 mL single-necked flask, add 4 mL of dioxane to dissolve it, add 2.93 mL of 4M hydrochloric acid dioxane and stir at room temperature, and monitor the reaction progress by thin layer chromatography. After the reaction is complete, the reaction solution is dried and purified by silica gel column chromatography (DCM: MeOH = 5: 1) to obtain 309 mg of white solid, crude product. LC-MS / ESI [M+H] + 250.20.

[0217] Synthesis of (E)-3-(4-(((2-(2-methyl-7-phenyl-1H-indol-3-yl)ethyl)amino)methyl)phenyl)-N-((tetrahydro-2H-pyran-2-yl)oxy)acrylamide (Compound 13-13f) (Step e)

[0218]

[0219] Take 2-(2-methyl-7-phenyl-1H-indol-3-yl)ethyl-1-amine (309mg, 1.23mmol) in a 100mL single-mouth flask, add 10mL DCM and 3mL methanol to dissolve it, add (E)-3-(4-formylphenyl)-N-((tetrahydro-2H-pyran-2-yl)oxy)acrylamide (408mg, 1.48mmol) and stir at room temperature for 30min, then add sodium cyanoborohydride (77mg, 1.23mmol), continue stirring, and monitor the reaction process by thin layer chromatography. After the reaction is complete, the reaction solution is spin-dried, and the reaction solution is extracted with DCM 3 times. The organic phase is washed once with water and saturated brine, and the aqueous phase is stripped with DCM, and the organic phase is combined. Purification by silica gel column chromatography (DCM: MeOH = 15: 1) gives 357mg of brown oil, with a yield of 56.7%.

[0220] Synthesis of (E)-N-hydroxy-3-(4-(((2-(2-methyl-7-phenyl-1H-indol-3-yl)ethyl)amino)methyl)phenyl)acrylamide (Compound 13) (Step f)

[0221]

[0222] (E)-3-(4-(((2-(2-methyl-7-phenyl-1H-indol-3-yl)ethyl)amino)methyl)phenyl)-N-((tetrahydro-2H-pyran-2-yl)oxy)acrylamide (357 mg, 0.70 mmol) was placed in a 100 mL single-necked flask, 5 mL of DCM was added to dissolve it, and dioxane hydrochloride (1.58 mL) was added. The mixture was stirred at room temperature to precipitate a solid. The reaction progress was monitored by thin layer chromatography. After the reaction was complete, the mixture was filtered, the solid was washed with DCM, and the solid was filtered again to collect the solid. 181 mg of a yellow solid was obtained, with a yield of 60.7%.

[0223] 1 H NMR (500 MHz, DMSO-d 6 )δ10.66(s,1H),9.72(s,2H),7.66(d,J=8.4Hz,2H),7.63-7.58(m,4H),7.53-7.48(m,4H),7.47-7.44(m,1H),7.42-7.38(m,1H) ,7.08-6.99(m,2H),6.58(d,J=15.8Hz,1H),4.20(t,J=5.9Hz,2H),3.14(dd,J=10.4,6.3Hz,2H),3.04-3.00(m,3H),2.34(s,3H). 13 C NMR (126 MHz, DMSO-d 6 )δ162.52,138.98,137.50,135.35,133.92,133.35,132.50,130.66(2C),128.88(2C),128.73,128.32(2C),127 .61(2C),127.08,124.61,120.65,120.07,119.07,116.70,105.98,49.39,46.89,20.57,11.15.LC-MS / ESI[M+H] + 426.20.HRMS(ESI):calcd for C 27 H 27 N 3 O 2 [M+H] + :(m / z):425.2103; found:426.2186.

[0224] Synthesis of (E)-3-(4-formyl-2-methylphenyl)acrylic acid (Compound 14-14b) (Step a)

[0225]

[0226] 4-Bromo-3-methylbenzaldehyde (500mg, 2.51mmol), acrylic acid (905mg, 12.56mmol), bisacetonitrile palladium dichloride (65mg, 0.25mmol), tri(o-methylphenyl)phosphine (152mg, 1.31mmol) and N,N-diisopropylethylamine (3.244g, 25.10mmol) were placed in a 50mL Schlenk tube, and then 10mL of ultra-dry n-butanol was added. The mixture was heated to 95°C and refluxed for about 3h under nitrogen protection. After the reaction was completed, the mixture was filtered with diatomaceous earth and the filtrate was collected. The reaction solution was extracted with EA 3 times, the organic phase was washed once with water and saturated brine, the aqueous phase was stripped with EA, the organic phases were combined, and purified by silica gel column chromatography (DCM: MeOH = 25: 1) to obtain 570mg of a yellow solid.

[0227] LC-MS / ESI[MH] - 189.10.

[0228] Synthesis of (E)-3-(4-formyl-2-methylphenyl)-N-((tetrahydro-2H-pyran-2-yl)oxy)acrylamide (Compound 14-14c) (Step b)

[0229]

[0230] (E)-3-(4-formyl-2-methylphenyl)acrylic acid (570 mg, 3.00 mmol) was placed in a 100 mL single-necked flask, 10 mL of DMF was added to dissolve it, HATU (1.141 g, 3.00 mmol) and DIPEA (775 mg, 6.00 mmol) were added, stirred evenly at room temperature, and O-(tetrahydro-2H-pyran-2-yl)hydroxylamine (351 mg, 3.00 mmol) was added, and stirring was continued. The reaction was completed in about 30 minutes. The reaction solution was extracted with EA 3 times, the organic phase was washed once with water and saturated brine, the aqueous phase was stripped with EA, the organic phases were combined, and purified by silica gel column chromatography (PE:EA=2:1) ​​to obtain 336 mg of yellow oily liquid, with a yield of 56.9%.

[0231] 1 H NMR(500MHz,Chloroform-d)δ10.00(s,1H),8.05-7.94(m,2H),7.80-7.71(m,1H),7.68-7.61(m,1H),7.57-7.41 (m,1H),4.04-3.94(m,1H),3.75-3.63(m,2H),2.52(s,3H),1.90-1.83(m,3H),1.67-1.61(m,3H).LC-MS / ESI[MH] - 288.10.

[0232] Synthesis of (E)-3-(2-methyl-4-(((2-(2-methyl-1H-indol-3-yl)ethyl)amino)methyl)phenyl)-N-((tetrahydro-2H-pyran-2-yl)oxy)acrylamide (Compound 14-14d) (Step c)

[0233]

[0234] Take 2-methylindole-3-ethylamine (100mg, 0.57mmol) in a 100mL single-necked flask, add 10mL DCM and 3mL methanol to dissolve it, add (E)-3-(4-formyl-2-methylphenyl)-N-((tetrahydro-2H-pyran-2-yl)oxy)acrylamide (138mg, 0.48mmol) and stir at room temperature for 30min, then add sodium triacetoxyborohydride (304mg, 1.43mmol), continue stirring, and monitor the reaction process by thin layer chromatography. After the reaction is complete, the reaction solution is spin-dried, and the reaction solution is extracted with DCM 3 times. The organic phase is washed once with water and saturated brine, and the aqueous phase is stripped with DCM, and the organic phase is combined. Purification by silica gel column chromatography (DCM: MeOH = 10: 1) obtains 204mg of a yellow oily liquid.

[0235] LC-MS / ESI[MH] - 448.20.

[0236] Synthesis of (E)-N-hydroxy-3-(2-methyl-4-(((2-(2-methyl-1H-indol-3-yl)ethyl)amino)methyl)phenyl)acrylamide (Compound 14) (Step d)

[0237]

[0238] (E)-3-(2-methyl-4-(((2-(2-methyl-1H-indol-3-yl)ethyl)amino)methyl)phenyl)-N-((tetrahydro-2H-pyran-2-yl)oxy)acrylamide (342 mg, 0.76 mmol) was placed in a 100 mL single-necked flask, 5 mL of DCM was added to dissolve it, 1.91 mL of dioxane hydrochloride (4M) was added, and the mixture was stirred at room temperature to precipitate a solid. The reaction progress was monitored by thin layer chromatography. After the reaction was complete, the mixture was filtered, the solid was washed with DCM, and the solid was filtered again to collect the solid. 160 mg of a yellow solid was obtained, with a yield of 57.4%.

[0239] 1 H NMR (500 MHz, DMSO-d 6)δ10.89(s,1H),9.56(s,2H),7.64(d,J=15.7Hz,1H),7.56(d,J=8.2Hz,1H),7.48-7.41(m,3H),7.24(d,J=8.0Hz,1H),7.01-6.95(m,1 H),6.95-6.90(m,1H),6.46(d,J=15.7Hz,1H),4.14(t,J=5.9Hz,2H),3.11-3.05(m,2H),3.01-2.95(m,2H),2.38(s,3H),2.33(s,3H). 13 C NMR (126 MHz, DMSO-d 6 )δ171.98,146.28,144.71,144.39,143.69,142.42,142.12,141.76,137.45,135.61,130.80,129 .69,127.78,126.71,120.03,114.71,75.84,58.78,56.32,29.99,28.93,20.66.LC-MS / ESI[M+H] + 364.20.

[0240] The synthetic route of target compounds 15-16 is similar to that of compound 8

[0241] Synthesis of (E)-N-hydroxy-3-(4-(((2-(7-phenyl-1H-indol-3-yl)ethyl)amino)methyl)phenyl)acrylamide (Compound 15)

[0242]

[0243] 60 mg of brown solid, yield 39.7%.

[0244] 1 H NMR (500 MHz, DMSO-d 6 )δ10.77(s,1H),7.62(d,J=7.5Hz,2H),7.57-7.45(m,7H),7.42-7.36(m,2H),7.15(s,1H),7.09(d,J=8. 2Hz,2H),6.57(d,J=15.9Hz,1H),3.95(s,2H),3.03(d,J=7.3Hz,2H),2.96(d,J=7.4Hz,2H),1.89(s,2H). 13 C NMR (126 MHz, DMSO-d 6)δ163.22,139.35,138.25,134.67,133.87,129.99(2C),129.44(2C),128.71(2C),128.59,127.93(2C),127 .66,125.75,124.31,121.76,119.80,119.53,118.30,112.29,51.64,48.84,24.19,22.07.LC-MS / ESI[M+H] + 412.20.

[0245] Synthesis of (E)-3-(4-(((2-(7-cyclopentyl-2-methyl-1H-indol-3-yl)ethyl)amino)methyl)phenyl)-N-hydroxyacrylamide (Compound 16)

[0246]

[0247] 94 mg of green solid, yield 99.1%.

[0248] 1 H NMR (500 MHz, DMSO-d 6 )δ10.70(s,1H),9.64(s,2H),7.65-7.59(m,4H),7.46(d,J=15.8Hz,1H),7. 29-7.23(m,1H),6.88-6.85(m,2H),6.57(d,J=15.9Hz,1H),4.18(t,J=5.9Hz ,2H),3.43-3.33(m,1H),3.10-3.06(m,2H),3.00-2.94(m,2H),2.34(s,3H), 2.08-2.02(m,2H),1.84-1.74(m,2H),1.72-1.65(m,2H),1.62-1.56(m,2H). 13 C NMR (126 MHz, DMSO-d 6 )δ162.51,137.50,135.34,134.17,133.32,132.41,130.63(2C),128.16,127.60(2C),120.06,118 .68,116.38,114.78,105.55,49.37,46.94,32.82(2C),24.93(2C),20.63,11.16.LC-MS / ESI[M+H] + 418.25.

[0249] Example 2. Biological activity test

[0250] Experimental procedures

[0251] (1) Transfer 50 nL of compound into a 384-well plate, with each well containing 2 replicates. Centrifuge the 384-well plate at 1000 RPM.

[0252] (2) Add 2.5uL HDAC to each test well and incubate at 25°C for 15 minutes.

[0253] (3) Add 2.5 μL of substrate to each reaction well and incubate at 37°C for 40 min.

[0254] (4) Add 5uL of developer mixture to each test well and incubate for 10 minutes.

[0255] (5) Use BMG to read the FI signal (em: 340; ex: 450 nm).

[0256] (6) “***” indicates IC 50 <100nM, "**" indicates 1μM>IC 50 >100nM, “*” indicates IC 50 >1μM.

[0257]

[0258]

[0259] The inventors used Panobinostat, an HDAC inhibitor that was withdrawn from the market by the FDA due to its high toxicity and side effects, as a lead compound. The biological experiments involved in the present invention found that the potential cause of Panobinostat's cardiotoxicity was off-target to S1PR2 and S1PR3. Based on this, through structural modification, the aim is to find HDAC inhibitors with activity equivalent to or better than the lead compound and low toxicity. The test results show that the IC values ​​of Compound 6, Compound 8, Compound 9, Compound 10 and Compound 13 for the inhibitory activity of HDAC1 and HDAC6 are 50 At nM level.

[0260] Example 3. S1PR biological activity test

[0261] Experimental Principle

[0262] Tango detection technology monitors ligand-dependent GPCR activity through the interaction between β-arrestin and GPCR. Tango cell S1PR2 and S1PR3 cell lines were constructed. Tango analysis detects the recruitment of β-arrestin, a reaction that occurs after G protein activation and G protein-coupled receptor kinase-guided phosphorylation of intracellular residues of GPCRs. The TangoTM system converts the β-arrestin recruitment signal into the expression of intracellular reporter genes. Various reporter genes have been used for in vitro and in vivo studies, including luciferase and β-lactamase. Luciferase has a shorter half-life than β-lactamase, providing some advantages, making it a common choice for screening tests. Therefore, the constructed Tango cells selected luciferase as the reporter gene and used Promega's Bright Glo TM The luciferase expression level was monitored by Luciferase Assay System.

[0263] Experimental Procedure

[0264] (1) Plating: 96-well white plate 8*12 plating density: 1.5*105 cells / mL plating volume per well: 70 μL.

[0265] (2) Cultivation: Culture in a CO2 incubator at 37°C overnight (16-20 h). The cell density is about 90% on the next day.

[0266] (3) Administration: 10 concentration gradients, 4 or 5-fold dilution, no compound added to the last well as a control, 2 or 3 replicates. Dissolve the compound in DMSO to ensure that the final DMSO concentration in each well is consistent (one thousandth). Add the compound of the corresponding concentration to a 96-well white plate, 30 μL per well. Add the compound to the wall without touching the cells at the bottom.

[0267] (4) Luciferase detection and analysis: After 20 hours of drug exposure, samples were collected and cells were removed from the incubator. TM Luciferase Assay System was used for luciferase analysis: 10 μL of reagent substrate was added to each well and the well was gently shaken for 15 min at room temperature in the dark. The well was placed on a microplate reader for reading, and the fluorescence relative light unit (RLU) was measured. The fluorescence (Luminescence) value was recorded, and the data was processed by Graphpad prism software to calculate the half effective concentration (EC) of the compound for S1PR2 and S1PR3. 50 )

[0268]

[0269]

[0270] The inventors used panobinostat, an HDAC inhibitor that was withdrawn from the market by the FDA due to its high toxicity and side effects, as a lead compound. The biological experiments involved in the present invention found that the potential cause of panobinostat's cardiotoxicity was off-target to S1PR2 and S1PR3. Based on this, through structural modification, the inventors aimed to find HDAC inhibitors with activities equivalent to or better than the lead compound, and lower agonist activity to S1PR2 and S1PR3 than the lead compound. The test results showed that the agonist activity of all compounds on S1PR2 and S1PR3 was lower than that of the lead compound, among which the IC values ​​of compound 6, compound 8, compound 9, compound 10 and compound 13 for the inhibitory activity on HDAC1 and HDAC6 were 50 At the nM level, this means that the safety of this compound is higher than that of the lead compound.

[0271] Example 4. Toxicity evaluation

[0272] 4.1 Zebrafish toxicity assessment

[0273] The drug concentrations were set to 0, 1.5, 3.5, 7.5, 15, 30, and 60 μM for toxicity evaluation. The zebrafish treated with the drug were loaded and fixed in the zebrafish chip, and the zebrafish heart rate was processed and recorded, and data was collected to measure the body length, pericardial ratio, and yolk sac of the zebrafish.

[0274] 4.2 Experimental Results

[0275] The experimental results are as follows Figure 1-5 As shown, panobinostat significantly decreased heart rate at 1.5 μM, and showed an increase in yolk sac and pericardium percentage at 7.5 μM. Compound 8 significantly decreased heart rate at 7.5 μM, increased yolk sac percentage at 15 μM, and no significant pericardium percentage; Compound 13 significantly decreased heart rate at 30 μM, and no significant pericardium percentage and yolk sac percentage. (Note: Compared with the control group, ns: no significant difference, *: p < 0.05, **: p < 0.01, ***: p < 0.001, ****: p < 0.0001;)

[0276] By testing the effect of compounds on zebrafish heart rate, it is possible to evaluate whether the compounds will cause cardiotoxicity. Figure 1As shown, panobinostat significantly reduced heart rate at 1.5 μM, and showed a clear trend of heart rate reduction as the dosage increased; while compound 8 had no significant effect on reducing heart rate at a dose of 3.5 μM, and only produced a heart rate reduction effect at a dose of 7.5 μM; similarly, compound 13 did not produce a heart rate reduction effect at a dose of 15 μM, and only produced a heart rate reduction effect at a dose of 30 μM. This result proves that compound 8 and compound 13 are much safer than panobinostat in terms of the effect on heart rate. (Note: Compared with the control group, ns: no significant difference, *: p<0.05, **: p<0.01, ***: p<0.001, ****: p<0.0001;)

[0277] By testing the effect of compounds on zebrafish body length, it is possible to evaluate whether the compounds affect zebrafish growth. Figure 2 As shown, panobinostat and compound 8 had no effect on the body length of zebrafish, while compound 13 at 7.5 μM could increase the body length of zebrafish, indicating that compound 13 could promote the growth of zebrafish. This result proves that compound 8 and compound 13 are as safe as panobinostat in affecting the body length of zebrafish. (Note: Compared with the control group, ns: no significant difference, *: p<0.05, **: p<0.01, ***: p<0.001, ****: p<0.0001;)

[0278] By testing the effect of compounds on the yolk sac ratio of zebrafish, it is possible to evaluate whether the compounds are toxic to the growth and development of zebrafish. Figure 3 As shown, panobinostat can affect the proportion of zebrafish yolk sac at 7.5μM and increase it, indicating that panobinostat has growth and development toxicity at 7.5μM, while compound 8 can affect the proportion of zebrafish yolk sac at 15μM, and compound 13 shows no effect at 60μM. The above results prove that compound 8 and compound 13 are safer than panobinostat in affecting the growth and development of zebrafish. (Note: Compared with the control group, ns: no significant difference, *: p<0.05, **: p<0.01, ***: p<0.001, ****: p<0.0001)

[0279] By testing the effect of compounds on the zebrafish pericardium ratio, the zebrafish cardiotoxicity of the compounds can be evaluated. Figure 4As shown, panobinostat can increase the proportion of zebrafish pericardium at 7.5μM, indicating that panobinostat has cardiotoxicity at 7.5μM, while compound 8 and compound 13 showed no effect at 30μM and 60μM, respectively. The above results prove that compound 8 and compound 13 are much higher than panobinostat in zebrafish cardiac safety. (Note: Compared with the control group, ns: no significant difference, *: p<0.05, **: p<0.01, ***: p<0.001, ****: p<0.0001)

[0280] 4.3 Experimental conclusion:

[0281] Compounds 8 and 13 maintained their original activity against HDAC, while reducing their activity against S1PR2 and S1PR3, thereby reducing potential toxicity. The above experimental results show that the toxicity of compound 8 is significantly reduced, and compound 13 is essentially non-toxic.

[0282] discuss:

[0283] After extensive and in-depth research, the inventors used Panobinostat, an HDAC inhibitor that was delisted by the FDA due to its high toxicity and side effects, as a lead compound. Based on the biological experiments involved in the present invention, it was found that the potential cause of Panobinostat's cardiotoxicity was off-target to S1PR2 and S1PR3. Through structural modification, a series of HDAC inhibitor small molecule compounds that have not been reported in the literature were designed and synthesized. The obtained compounds were tested for S1PR2 and S1PR3 activity. The test results showed that all compounds had lower agonist activity to S1PR2 and S1PR3 than the lead compound. At the same time, their kinase activity was tested, and a group of compounds that could inhibit HDAC1 and HDAC6 and had weak activity to S1PR2 and S1PR3 were obtained. The zebrafish toxicity evaluation of the HDAC1 / 6 inhibitor with excellent activity was carried out, and it was found that it had low toxicity and had a certain development potential compared with Panobinostat.

[0284] The above results indicate that as HDAC inhibitors, compounds 8 and 13, especially compound 13, are comparable to panobinostat at the kinase level and are less toxic than panobinostat.

[0285] The above work lays the foundation for the treatment of HDAC-mediated diseases.

[0286] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A compound of formula I, or a pharmaceutically acceptable salt, isomer, racemate, precursor or solvate thereof: In the formula, R 1 is selected from the group consisting of: hydrogen, substituted or unsubstituted C1-C10 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C3-C6 cycloalkenyl, substituted or unsubstituted C3-C8 lactone, substituted or unsubstituted C1-C10 amide, substituted or unsubstituted C1-C10 amide, substituted or unsubstituted C5-C10 aryl, substituted or unsubstituted C3-C8 heterocyclyl, substituted or unsubstituted C5-C10 aromatic heterocyclyl having 1 to 3 heteroatoms independently selected from N, O or S; R 2 Selected from: hydrogen, halogen, cyano, Ring A is selected from: a substituted or unsubstituted C5-8 aryl group, a substituted or unsubstituted C5-C10 heteroaryl group having 1 to 3 heteroatoms independently selected from N, O or S, a substituted or unsubstituted C5-C10 heterocyclyl group having 1 to 3 heteroatoms independently selected from N, O or S, a substituted or unsubstituted C5-8 aryl group, or a C5-8 heteroaryl group having 1 or 2 heteroatoms independently selected from N, O or S and a substituted or unsubstituted C5-8 heterocyclyl group having 1 or 2 heteroatoms independently selected from N, O or S.

2. The compound according to claim 1, or a pharmaceutically acceptable salt, isomer, racemate, precursor or solvate thereof, characterized in that: In the formula, R 1 Selected from: hydrogen, substituted or unsubstituted C1-C6 alkyl; R 2 Selected from: Ring A is selected from: substituted or unsubstituted C5-8 aryl, substituted or unsubstituted C5-8 aryl and substituted or unsubstituted C5-8 heterocyclic group having 1 or 2 heteroatoms independently selected from N, O or S.

3. The compound according to claim 2, or a pharmaceutically acceptable salt, isomer, racemate, precursor or solvate thereof, characterized in that: In the formula, R 1 Selected from: hydrogen; R 2 Selected from: Ring A is selected from: substituted or unsubstituted phenyl, X is selected from: N, O, S, CH; R 3 Selected from: hydrogen, substituted or unsubstituted C1-3 alkyl, substituted or unsubstituted C1-3 alkoxy; R 4 Selected from: hydrogen, substituted or unsubstituted C5-8 aryl, substituted or unsubstituted C5-8 cycloalkyl; R5 is selected from: hydrogen, substituted or unsubstituted C1-3 alkyl, substituted or unsubstituted benzyl; R 6 Selected from: hydrogen, substituted or unsubstituted C1-3 alkyl.

4. The compound according to claim 3, or a pharmaceutically acceptable salt, isomer, racemate, precursor or solvate thereof, characterized in that: In the formula, R 1 Selected from: hydrogen; R 2 Selected from: Ring A is selected from: X is selected from: CH; R 3 Selected from: hydrogen, trifluoromethoxy; R 4 is selected from: hydrogen, substituted or unsubstituted phenyl; R5 is selected from the group consisting of: hydrogen, substituted or unsubstituted methyl, substituted or unsubstituted benzyl; R 6 Selected from: hydrogen, substituted or unsubstituted methyl.

5. A compound selected from the group consisting of, or a pharmaceutically acceptable salt, isomer, racemate, precursor or solvate thereof, 6. The compound according to claim 5, or a pharmaceutically acceptable salt, isomer, racemate, precursor or solvate thereof, characterized in that: The compound is selected from the group consisting of: Preferably, the compound is selected from the group consisting of:

7. A pharmaceutical composition comprising the compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt, isomer, racemate, precursor or solvate thereof, and a pharmaceutically acceptable excipient.

8. Use of the compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt, isomer, racemate, precursor or solvate thereof in the preparation of an HDAC inhibitor.

9. The use according to claim 8, characterized in that The HDAC inhibitor is a drug for treating diseases related to abnormal regulation of histone deacetylase.

10. The use according to claim 9, characterized in that The disease associated with abnormal regulation of histone deacetylase is cancer, neurodegenerative disease, AIDS, Alzheimer's disease, malaria or diabetes.

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