Benzothiazepine compound, preparation method therefor, and use thereof

Abstract

Disclosed are a benzothiazepine compound, a preparation method therefor, and use thereof. Specifically disclosed is a compound represented by formula (I) or a pharmaceutically acceptable salt thereof. The compound or the salt thereof has one or more of the following advantages: selective inhibition of HDAC6, stable metabolism, low toxicity, inhibition of tumor cells, protection of nerve cells, and having a relatively good in vivo antidepressant effect, and is expected to be used as a novel therapeutic agent for the clinical treatment of cancer or diseases of the nervous system.

Description

Benzothiazepine compounds, preparation method and use thereof

[0001] This application claims priority to Chinese patent application 2024108196682 with a filing date of 2024 / 6 / 24. This application incorporates the entirety of the aforementioned Chinese patent application. TECHNICAL FIELD

[0002] The present application relates to a benzothiazepine compound, a preparation method and use thereof. BACKGROUND

[0003] Histone deacetylases (HDACs) and histone acetyltransferases (HATs) regulate gene expression by adjusting the level of histone acetylation in the nucleus. Currently, there are 18 subtypes of mammalian HDACs, which are divided into four categories: class I (HDAC1, HDAC2, HDAC3, HDAC8); class II is divided into two subfamilies IIa (HDAC4, HDAC5, HDAC7, HDAC9) and IIb (HDAC6, HDAC10); class III (Sirt1-Sirt7); class IV (HDAC11). The inhibitory effect of histone deacetylase inhibitors (HDACi) on tumor cell migration, invasion, metastasis and anti-tumor angiogenesis has been confirmed. Currently, there are five HDACi on the market as anti-tumor drugs, namely vorinostat, belinostat, panobinostat, romidepsin and chidamide, the former three are broad-spectrum inhibitors, and the latter two selectively act on class I subtypes, but also have certain side effects.

[0004] Currently, HDACs subtype selective inhibitors have become a research hotspot in this field, especially HDAC6 subtype inhibitors have attracted much attention. HDAC6 is involved in a wide range of disease areas, including tumors, nervous system diseases, inflammation, autoimmune response and bacterial infection, etc. Research has found that HDAC6 gene knockout mice survive well and develop normally, indicating that selective inhibition of HDAC6 may not cause serious toxic side effects.

[0005] As of March 2024, there are 3 HDAC6 inhibitors in clinical phase 2 research: Citarinostat (indication: multiple myeloma), CKD-506 (indication: autoimmune diseases), and Rocilinostat (indication: diabetic peripheral neuropathy, refractory plasma cell myeloma, relapsed multiple myeloma, breast cancer, peripheral nervous system diseases, relapsed chronic lymphocytic leukemia); 3 drugs are in clinical phase 1 research: CKD-510 (indication: peroneal muscle atrophy, rare diseases), TYA-11631 (indication: heart failure, dilated cardiomyopathy), CKD-504 (indication: Huntington's disease), and a large number of HDAC6 inhibitors are in preclinical development.

[0006] Patent 202010304883.0 discloses a class of benzazepine compounds, among which compound T-1 has an IC 50 = 37nM, which is weaker than SW-100 (IC 50 = 2.4nM) by nearly one order of magnitude. At the same time, the substituents on the benzene ring have a greater impact on activity, for example, compound T-5 has an IC 50 = 5.6nM, which is better than T-1. In addition, the heterocycle of benzazepine also has a significant impact on activity, for example, the activity of benzothiazepine T-16 is better than that of benzooxazepine compound T-15.

[0007] The above publications and patents suggest that the difference in heterocyclic structure containing nitrogen atoms and the difference in substituents on the benzene ring in the formula affect the HDAC6 inhibition activity of the compounds, but the degree of influence and the resulting pharmacological effects cannot be predicted and need to be verified by experiments. In view of the importance of HDAC6 inhibitors, there is an urgent need to develop HDAC6 inhibitors with novel structure, good metabolic properties, and high selectivity. SUMMARY

[0008] The technical problem to be solved by the present application is to overcome the defects of low selectivity or poor drugability (such as metabolic instability) of HDAC6 inhibitors in the prior art, and to provide a class of benzothiazepine compounds, their preparation methods and uses. The compounds of the present application have one or more of the following advantages: HDAC6 selective inhibition, metabolic stability, low toxicity, inhibition of tumor cells, protection of nerve cells, good in vivo anti-depression effect, and are expected to be used as new cancer or nervous system disease treatment agents for clinical use.

[0009] The present application provides a compound as shown in formula (I) or a pharmaceutically acceptable salt thereof:

[0010] wherein,

[0011] X1is

[0012] R 1a , R 1b , R 1c , and R 1d are independently H, deuterium, halogen, cyano, hydroxyl, or R 1 -L-; and when X1is R 1a , R 1b , R 1c , R 1d , R 2a , R 2b , R 2c , and R 2d are not simultaneously hydrogen;

[0013] -L- is independently a bond, -(Ci-C4alkyl)-, -0-, -C(=0)-, or -S(=0)2-;

[0014] R 1 is independently Ci-C6alkyl, C3-C7cycloalkyl, C6-C 10 aryl, (R a R b )N-, 3-7 membered heterocycloalkyl, or 5-10 membered heteroaryl, or, said Ci-C6alkyl, C3-C7cycloalkyl, C6-C 10 aryl, (R a R b )N-, 3-7 membered heterocycloalkyl, and 5-10 membered heteroaryl are each independently substituted with one or more substituents R c ; in said 3-7 membered heterocycloalkyl and 3-7 membered heterocycloalkyl within said 3-7 membered heterocycloalkyl substituted with one or more substituents R c , the heteroatom group is selected from one or more of N, O, S, S(=0), and S(=0)2, and the number of heteroatoms is 1-3; in said 5-10 membered heteroaryl and 5-10 membered heteroaryl within said 5-10 membered heteroaryl substituted with one or more substituents R c , the heteroatom group is selected from one or more of N, O, and S, and the number of heteroatoms is 1-4; R c is independently hydroxyl, halogen, cyano, or Ci-C4alkyl; when there is more than one substituent, they are the same or different;

[0015] R a and R b are each independently hydrogen or Ci-C4alkyl;

[0016] R 2a , R 2b , R 2cand R 2d independently hydrogen, deuterium, halogen, C1-C4alkyl or C1-C4alkyl substituted with one or more halogens; when halogen is multiple, same or different.

[0017] In certain preferred embodiments of the application, certain groups of the compounds of Formula (I) or a pharmaceutically acceptable salt thereof are defined as follows, and the groups not mentioned are as described in any of the embodiments of the application (simply referred to as "in an embodiment of the application"). The same abbreviation is used for other embodiments of the application, and will not be repeated.

[0018] In an embodiment of the application, R 1a , R 1b , R 1c , R 1d , R 2a , R 2b , R 2c and R 2d , the halogen is F, Cl, Br or I.

[0019] In an embodiment of the application, R 1 , the C1-C6alkyl is C1-C3alkyl, for example methyl.

[0020] In an embodiment of the application, X1is preferably

[0021] In an embodiment of the application, R 1a is H.

[0022] In an embodiment of the application, R 1b and R 1c are independently H, halogen or R 1 -L-; -L- is a bond or -O-, R 1 is C1-C6alkyl.

[0023] In an embodiment of the application, R 1b is H, F, Cl, Br, methyl or methoxy.

[0024] In an embodiment of the application, R 1c is H or halogen.

[0025] In an embodiment of the application, R 1c is H, F or Cl.

[0026] In an embodiment of the application, R 1d is H.

[0027] In an embodiment of the application, R 2a , R2b R 2c and R 2d are independently hydrogen or halogen.

[0028] In one embodiment of the application, R 2a , R 2b , R 2c and R 2d are independently hydrogen or F.

[0029] In one embodiment of the application, R 1b and R 1c are independently halogen or R 1 -L-.

[0030] In one embodiment of the application, R 1b and R 1c are independently halogen or R 1 -L-, R 1a and R 1d are H.

[0031] In one embodiment of the application,

[0032] wherein, in A and B, R 1b and R 1c are independently halogen, cyano, hydroxy or R 1 -L-;

[0033] -L- is independently a bond, -(C1-C4alkyl)-, -O-, -C(=O)- or -S(=O)2-;

[0034] R 1 is independently C1-C6alkyl, C3-C7cycloalkyl, C6-C 10 aryl, (R a R b )N-, 3-7 membered heterocycloalkyl or 5-10 membered heteroaryl, or, said C1-C6alkyl, C3-C7cycloalkyl, C6-C 10 aryl, (R a R b )N-, 3-7 membered heterocycloalkyl and 5-10 membered heteroaryl are each independently substituted with one or more substituents R c ; in said 3-7 membered heterocycloalkyl and 3-7 membered heterocycloalkyl within said 3-7 membered heterocycloalkyl substituted with one or more substituents R c , the heteroatom group is selected from one or more of N, O, S, S(=O) and S(=O)2, and the number of heteroatoms is 1-3; and said 5-10 membered heteroaryl and 5-10 membered heteroaryl substituted with one or more substituents R c ​In the 5-10 membered heteroaryl group of the substituted 5-10 membered heteroaryl group, the heteroatom group is selected from one or more of N, O and S, and the number of heteroatoms is 1-4; R c independently hydroxyl, halogen, cyano or C1-C4 alkyl; when there are multiple substituents, they are the same or different;

[0035] R a and R b are each independently hydrogen or C1-C4 alkyl;

[0036] Preferably, in A and B, R 1b and R 1c are independently halogen (e.g., F, Cl or Br) or R 1 -L-;

[0037] -L- is independently a bond or -O-;

[0038] R 1 is independently C1-C6 alkyl, preferably C1-C3 alkyl, e.g., methyl;

[0039] wherein, in C, D, E and F, R 1b and R 1c are as defined in any of the preceding aspects.

[0040] In one aspect of the application, the compound of formula (I) or a pharmaceutically acceptable salt thereof,

[0041] -L- is independently a bond or -O-;

[0042] R 1 is independently C1-C6 alkyl;

[0043] R 2a , R 2b , R 2c and R 2d are independently hydrogen or halogen;

[0044] when X1is , R 1a is hydrogen, R 1b and R 1c are independently halogen or R 1 -L-; R 1d is H, halogen or R 1 -L-;

[0045] when X1is , R 1a , R 1b , R 1c and R 1d are independently H, halogen or R 1 -L-.

[0046] In one embodiment of the present application, the compound of formula (I) has the following structure:

[0047] wherein R 2a and R 2d are independently hydrogen or halogen; the halogen is preferably F;

[0048] X1and R 1b are defined as any one of the following cases:

[0049] Case 1: X1is R 1b is halogen or R 1 -L-; -L- is -O-; R 1 is independently C1-C6 alkyl, preferably C1-C3 alkyl, such as methyl;

[0050] Case 2: X1is R 1b is H, halogen or R 1 -L-; -L- is -O-; R 1 is independently C1-C6 alkyl, preferably C1-C3 alkyl, such as methyl.

[0051] In one embodiment of the present application, the compound of formula (I) has the following structure:

[0052] wherein X1is X1is preferably

[0053] R 1b is halogen (e.g. F or Br) or R 1 -L-; -L- is independently -O-; R 1 is independently C1-C6 alkyl, preferably C1-C3 alkyl, such as methyl; R 1b is preferably halogen.

[0054] In one embodiment of the present application, the compound of formula (I) is any one of the following structures:

[0055] The present application also provides a method for preparing a compound of formula (I), characterized in that it comprises the following steps:

[0056] In a solvent, in the presence of a base, subjecting a compound of formula (II) to a hydroxylamine reaction as shown below with hydroxylamine to obtain the compound of formula (I):

[0057] wherein X1, R 1a , R 1b , R 1c , R 1d , R 2a , R 2b , R 2c and R 2d are as defined in any of the preceding embodiments;

[0058] R 3 is H or C1-C6 alkyl.

[0059] In an embodiment of the present application, the solvent is an alcohol solvent, such as methanol.

[0060] In an embodiment of the present application, the base is an alkali metal hydroxide, such as KOH.

[0061] In an embodiment of the present application, R 3 is hydrogen, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, sec-butyl or t-butyl.

[0062] In an embodiment of the present application, the mass / volume ratio of the compound of formula (II) to the solvent is 0.1 g / L to 20 g / L.

[0063] In an embodiment of the present application, the molar ratio of the compound of formula (II) to the hydroxylamine is 1:5 to 1:50, such as 1:10.

[0064] In an embodiment of the present application, the temperature of the hydroxylamine reaction is room temperature to 80°C; preferably 25-80°C.

[0065] In an embodiment of the present application, the method for preparing the compound of formula (I) further comprises a post-treatment; the post-treatment comprises one or more of the following steps: after the hydroxylamine reaction, adjusting the pH to 6-7, filtering to obtain the solid, recrystallization, and the like.

[0066] In an embodiment of the present application, the method for preparing the compound of formula (I) further comprises the following step: in the presence of a base, a substitution reaction is carried out between the compound of formula (III) and the compound of formula (IV) in a solvent to obtain the compound of formula (II).

[0067] wherein X1, R 1a , R 1b , R 1c , R 1d , R 2a , R 2bR 2c R 2d R 3 The definitions of X1, R 1a R 1b R 1c R 1d R 2a R 2b R 2c R 2d are as described in any of the preceding embodiments; and X is halogen, such as Br.

[0068] In an embodiment of the present application, in the substitution reaction, the solvent is a nitrile solvent, such as acetonitrile.

[0069] In an embodiment of the present application, in the substitution reaction, the base is an alkali metal carbonate, such as K2CO3.

[0070] The present application also provides a compound of formula (II) or a pharmaceutically acceptable salt thereof,

[0071] wherein the definitions of X1, R 1a R 1b R 1c R 1d R 2a R 2b R 2c R 2d are as described in any of the preceding embodiments.

[0072] R 3 is H or C1-C6 alkyl.

[0073] In an embodiment of the present application, the compound of formula (II) is any of the following structures:

[0074] The present application also provides a pharmaceutical composition comprising the compound of formula (I) or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers.

[0075] The present application also provides use of the compound of formula (I), a pharmaceutically acceptable salt thereof or the pharmaceutical composition in the preparation of an HDAC inhibitor or a medicament.

[0076] The HDAC inhibitor is preferably an HDAC6 inhibitor.

[0077] The medicament is preferably a medicament for preventing and / or treating cancer or a nervous system disease or a medicament for preventing and / or treating a disease or condition associated with HDAC.

[0078] The cancer is preferably pituitary tumor, ovarian cancer, colon cancer, breast cancer, liver cancer, pancreatic cancer, gallbladder cancer, gastrointestinal cancer, head and neck cancer, cervical cancer, prostate cancer, lung cancer, melanoma, germ cell tumor, gestational trophoblastic tumor, glioblastoma, myeloma, neuroblastoma-derived CNS tumor, monocyte leukemia, B-cell leukemia, T-cell leukemia, B-cell lymphoma, T-cell lymphoma, or mast cell-derived tumor; more preferably glioblastoma, myeloma, pituitary tumor, or lung cancer.

[0079] The nervous system disease is preferably stroke, depression, neurodegenerative disease, or peripheral neuropathy.

[0080] The stroke is preferably ischemic stroke or hemorrhagic stroke.

[0081] The depression is preferably major depressive disorder, persistent depressive disorder, bipolar disorder, seasonal affective disorder, or postpartum depression.

[0082] The neurodegenerative disease is preferably Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, spinocerebellar degeneration, Cushing's syndrome, or Rett syndrome.

[0083] The peripheral neuropathy is preferably Charcot-Marie-Tooth disease or giant axonal neuropathy.

[0084] In one aspect of the present application, the disease or disorder associated with HDAC is cancer or nervous system disease.

[0085] The cancer is preferably pituitary tumor, ovarian cancer, colon cancer, breast cancer, liver cancer, pancreatic cancer, gallbladder cancer, gastrointestinal cancer, head and neck cancer, cervical cancer, prostate cancer, lung cancer, melanoma, germ cell tumor, gestational trophoblastic tumor, glioblastoma, myeloma, neuroblastoma-derived CNS tumor, monocyte leukemia, B-cell leukemia, T-cell leukemia, B-cell lymphoma, T-cell lymphoma, or mast cell-derived tumor; more preferably glioblastoma, myeloma, pituitary tumor, or lung cancer.

[0086] The nervous system disease is preferably stroke, depression, neurodegenerative disease, or peripheral neuropathy.

[0087] The stroke is preferably ischemic stroke or hemorrhagic stroke.

[0088] The depression is preferably major depressive disorder, persistent depressive disorder, bipolar disorder, seasonal affective disorder, or postpartum depression.

[0089] The neurodegenerative disease is preferably Alzheimer's disease, Parkinson's disease, Huntington's chorea, amyotrophic lateral sclerosis, spinocerebellar degeneration, Cushing's syndrome, or Rett's syndrome.

[0090] The peripheral neuropathy is preferably Charcot-Marie-Tooth disease or giant axonal neuropathy.

[0091] The present application also provides use of a compound V having the following structure in the manufacture of a medicament for the prophylaxis and / or treatment of depression; the depression is preferably major depressive disorder, persistent depressive disorder, bipolar disorder, seasonal affective disorder, or postpartum depression.

[0092] Unless otherwise defined, each term used herein can be defined as follows:

[0093] When one of the variables is a bond, it indicates that the two groups to which it is attached are directly connected.

[0094] Herein, a substituent group used can be preceded by a single dash "-", indicating that the named substituent group is attached to the parent moiety through a single bond.

[0095] The term "halogen" means F, Cl, Br, I.

[0096] The term "alkyl" means a straight or branched chain alkyl group having the specified number of carbon atoms (e.g., C1-C6). Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, t-butyl, i-butyl, sec-butyl, n-pentyl, n-hexyl, and the like.

[0097] The term "aryl" means a cyclic group having the specified number of carbon atoms (e.g., C6-C 10 ) that is aromatic, which is monocyclic or polycyclic (e.g., 2 or 3). The aromatic ring includes, but is not limited to, benzene, naphthalene, and the like.

[0098] The term "heterocycloalkyl" means a saturated cyclic group having the specified number of ring atoms (e.g., 3-6 membered), the specified number of heteroatoms (e.g., 1 or 2), and the specified number of heteroatom species (1, 2, or 3 of N, O, S, S(=O), and S(=O)2). Examples of heterocycloalkyl groups include, but are not limited to, oxiranyl, aziridinyl, oxetanyl, azetidinyl, pyrrolinyl, tetrahydrofuranyl, tetrahydropyranyl, piperidinyl, morpholinyl, piperazinyl, and the like.

[0099] The term "heteroaromatic ring" refers to a cyclic group having a specified number of ring atoms (e.g., 5-10 membered), a specified number of heteroatoms (e.g., 1, 2, or 3), a specified type of heteroatom (one or more of N, O, and S), aromaticity, and being monocyclic or polycyclic. Heteroaromatic rings include, but are not limited to, pyridine rings and the like; the term "heteroaryl" refers to the structure resulting from the loss of one or more hydrogens from the definition of the term "heteroaromatic ring."

[0100] The term "pharmaceutically acceptable salt" means a salt of a compound of the present application that is found to possess the specific substituent groups discovered in the present application, which is prepared from a relatively non-toxic acid or base. Alkali addition salts can be prepared from the neutral form of the compounds of the present application by contacting the neutral form with a sufficient amount of the desired alkali to produce the salt in pure form. Acid addition salts can be prepared by contacting the neutral form of the compounds of the present application with a sufficient amount of the desired acid to produce the salt in pure form.

[0101] The term "pharmaceutically acceptable carrier" means all substances other than the active pharmaceutical ingredient that are included in a pharmaceutical preparation, which are generally classified into excipients and additives.

[0102] In the present application, the "inhibitors" can be used in vivo in mammalian organisms; they can also be used in vitro, mainly as experimental purposes, for example, as standard or control samples to provide a comparison, or in the form of kits according to the methods commonly used in the art.

[0103] The term "treatment" refers to therapeutic treatment. When used in reference to a specific disease or condition, treatment refers to: (1) relieving one or more of the signs or symptoms of the disease or condition, (2) interfering with (a) one or more of the biological cascades leading to or resulting in the condition or (b) one or more of the biological manifestations of the condition, (3) ameliorating one or more symptoms, effects, or side effects associated with the condition or one or more symptoms, effects, or side effects associated with treatment of the condition, or (4) slowing the development of the condition or one or more of the biological manifestations of the condition.

[0104] The term "prevention" refers to maintaining health in a subject with respect to a disease or condition referred to herein for a period of time. It will be appreciated that the period of time depends on the amount of pharmaceutical compound that has been administered and the individual factors of the subject discussed elsewhere in the specification. It will be appreciated that prevention can not be effective in all subjects treated with a compound according to the application. However, the term requires preferably that a statistically significant portion of the cohort or population of subjects is prevented from suffering from the disease or condition referred to herein or its attendant symptoms. Preferably, in this case, a group or population of subjects is envisioned that would have developed the disease or condition referred to herein in the absence of the preventive measures according to the application. Whether a portion is statistically significant can be readily determined by the skilled person using various well-known statistical evaluation tools discussed elsewhere in the specification.

[0105] On the basis of common general knowledge in the art, the above-mentioned preferred conditions can be combined arbitrarily, i.e. to obtain each preferred embodiment of the application.

[0106] The reagents and starting materials used in the present application are commercially available.

[0107] The positive progress effect of the present application is that pharmacological experiments show that the benzothiazepine compounds described in the present application have the following beneficial effects:

[0108] 1) The compounds of the present application show strong inhibitory activity against HDAC6 (IC 50 <10 nM), and some compounds have inhibitory activity comparable to or better than the positive control Rocilinostat (ACY-1215), SW-100. With respect to HDAC1, the compounds of the present application can selectively inhibit HDAC6 with a selectivity of > 100 times, which is significantly better than Rocilinostat and SW-100.

[0109] 2) The compounds of the present application all have certain protective effects against L-glutamic acid-induced damage to SH-SY5Y cells, suggesting that the compounds of the present application have neuroprotective effects and can be used for the treatment of stroke or neurodegeneration-related diseases.

[0110] 3) The compounds of the present application show good in vitro anti-tumor cell proliferation activity against a variety of tumor cells, and have high inhibitory activity against brain glioma cell line U-87MG, myeloma cell line RPMI8226, AtT-20 mouse pituitary tumor cells and human lung cancer cell line A549, and weak inhibitory activity against MRC-5 human normal embryonic lung fibroblasts, and have lower toxic side effects, revealing that the compounds of the present application can have lower toxic side effects when used as anti-tumor drugs.

[0111] 4), the compound of the present application is more stable in rat liver microsomes than the compound T-16* of the patent 202010304883.0, and has better PK properties.

[0112] 5) The hERG experiment results show that the inhibitory activity of the compound of the present application on the hERG potassium ion channel is greater than 10 μM, indicating that the potential cardiac toxicity of the compound of the present application is low.

[0113] 6) The mouse tail suspension experiment shows that the compound of the present application shows certain antidepressant activity, and the antidepressant activity is better than that of T-16*. BRIEF DESCRIPTION OF DRAWINGS

[0114] Figure 1 is the immobility time determination within 6 minutes of the mouse tail suspension experiment;

[0115] Figure 2 is the determination of the immobility time of forced swimming of rats. DETAILED DESCRIPTION

[0116] The present application will be further described below by way of examples, but the present application is not limited in the scope of the examples. The experimental methods in the following examples without specific conditions are selected according to conventional methods and conditions, or according to the instructions of the commodity.

[0117] The compound of the present application can be prepared by the following synthetic method:

[0118] The raw materials, compounds and reagents used in the above preparation method can be purchased through commercial channels.

[0119] Example 1: Preparation of compound T-1

[0120] The compound T-1 is prepared according to the synthetic method, and the synthetic route is as follows:

[0121] The specific synthesis operation is as follows:

[0122] 1) 3-bromopropionic acid (25 mmol, 1 eq) and K2CO3 (25 mmol, 1 eq) were dissolved in 10 mL of ice water, and the solution was added to a 10 mL aqueous solution of 3-fluorothiophenol (25 mmol, 1 eq) and NaOH (25 mmol, 1 eq), and the reaction was stirred under reflux. After the raw material was completely reacted by TLC monitoring, the reaction solution was cooled to room temperature, adjusted to pH ≈ 1 with 1N HCl, extracted with ethyl acetate, and the organic phase was washed with saturated NaCl solution, dried over anhydrous Na2SO4, and concentrated to obtain the intermediate 2a, which was directly subjected to the next step reaction.

[0123] 2) The above obtained intermediate 2a was mixed with 10 mL of concentrated sulfuric acid and stirred at room temperature overnight. Next day, the reaction was diluted with ice water and extracted with ethyl acetate. The organic layer was washed with water, saturated NaHC03 and saturated NaCl solution, dried over anhydrous Na2S04 and concentrated to get the intermediate 3a.

[0124] 3) Intermediate 3a (10 mmol, 1 eq) and hydroxylamine hydrochloride (20 mmol, 2 eq) were taken in 15 mL of pyridine and stirred at room temperature. The reaction was monitored by TLC and upon completion, the pyridine was removed under reduced pressure to get a light yellow oil. The oil was taken in water and stirred to get a solid which was filtered and dried to get the intermediate 4a.

[0125] 4) Intermediate 4a (10 mmol, 1 eq) was dissolved in 60 mL of dichloromethane and at 0 °C, DIBALH (1.0 M in Hexane, 60 mmol, 6 eq) was added drop wise to the above solution. The reaction mixture was allowed to stir at room temperature overnight. Next day, at 0 °C, NaF (120 mmol, 12 eq) was added and stirred and the reaction was quenched by slow addition of water. The aluminum salt came out as a gel from the reaction mixture which was further stirred at room temperature for about 3-5 h. The mixture was filtered (celite assisted filtration) and the filtrate was washed with saturated NaCl solution, dried over anhydrous Na2S04 and concentrated to get a light yellow oil which was purified by column chromatography (PE:EA = 100:1) to get the benzothiazepine intermediate 5a.

[0126] 5) Intermediate 5a (10 mmol, 1 eq) and K2C03 (15 mmol, 1.5 eq) were dissolved in 30 mL of acetonitrile and stirred at room temperature for 30 min. Then, p-bromomethyl benzoic acid methyl ester (10.5 mmol, 1.05 eq) was added and the reaction mixture was heated to reflux and stirred. The reaction was monitored by TLC and upon completion, the reaction mixture was cooled to room temperature and 30 mL of water was added to the reaction mixture which became turbid upon stirring. The reaction mixture was extracted with ethyl acetate and purified by column chromatography to get the intermediate 6a. 1 H NMR (400 MHz, Methanol-d4) δ 8.00 - 7.95 (m, 2H), 7.58 - 7.53 (m, 2H), 7.10 (dd, J = 8.8, 3.1 Hz, 1H), 6.97 (dd, J = 8.9, 5.2 Hz, 1H), 6.84 (ddd, J = 8.9, 7.9, 3.1 Hz, 1H), 4.45 (s, 2H), 3.90 (s, 3H), 3.22 - 3.14 (m, 2H), 2.93 - 2.85 (m, 2H), 1.96 - 1.87 (m, 2H); ESI-MS (m / z): 332.20 [M+H] + .

[0127] 6) Intermediate ester 6a (2 mmol, 1 eq) was added to 14 mL of freshly prepared hydroxylamine / KOH solution in methanol (20 mmol, 10 eq), and the reaction was stirred in an ice bath for about 1-2 hours. TLC monitoring showed that the reaction was complete, 1 N HCl was used to adjust the pH of the reaction solution to about 6-7, the reaction solution precipitated solids, which were filtered and recrystallized with methanol / water to obtain the target compound T-1. ESI-MS (m / z): 333.20 [M+H] +1 H NMR (400 MHz, DMSO-d6) δ 11.22 (s, 1H), 9.07 (s, 1H), 7.75 (d, J = 7.8 Hz, 2H), 7.41 (d, J = 7.8 Hz, 2H), 7.25 (d, J = 2.5 Hz, 1H), 7.06 (dd, J = 8.7, 2.7 Hz, 1H), 6.82 (d, J = 8.7 Hz, 1H), 4.43 (s, 2H), 3.39 (d, J = 44.2 Hz, 2H), 2.97 (t, J = 5.8 Hz, 2H), 1.85 (m, 2H); 13 C NMR (101 MHz, DMSO-d6) δ 164.57, 150.85, 142.58, 131.94, 130.98, 128.43, 127.96, 127.50, 127.38, 124.22, 120.35, 57.05, 52.22, 30.58, 28.81.

[0128] Example 2: Preparation of compound T-2

[0129] Compound T-2 was prepared according to the synthetic method of Example 1. ESI-MS (m / z): 349.10 [M+H]+; 1 H NMR (400 MHz, DMSO-d6) δ 11.22 (s, 1H), 9.07 (s, 1H), 7.75 (d, J = 7.8 Hz, 2H), 7.41 (d, J = 7.8 Hz, 2H), 7.25 (d, J = 2.5 Hz, 1H), 7.06 (dd, J = 8.7, 2.7 Hz, 1H), 6.82 (d, J = 8.7 Hz, 1H), 4.43 (s, 2H), 3.39 (d, J = 44.2 Hz, 2H), 2.97 (t, J = 5.8 Hz, 2H), 1.85 (m, 2H); 13 C NMR (101 MHz, DMSO-d6) δ 164.57, 150.85, 142.58, 131.94, 130.98, 128.43, 127.96, 127.50, 127.38, 124.22, 120.35, 57.05, 52.22, 30.58, 28.81.

[0130] Example 3: Preparation of compound T-3

[0131] Compound T-3 was prepared according to the synthetic procedure of Example 1. ESI-MS (m / z): 394.10 [M+H] + ; 1 H NMR (600 MHz, DMSO-d6) δ 11.15 (s, 1H), 9.02 (s, 1H), 7.71 (d, J = 8.1 Hz, 2H), 7.39 (d, J = 8.1 Hz, 2H), 7.36 (d, J = 2.5 Hz, 1H), 7.18 (dd, J = 8.7, 2.5 Hz, 1H), 6.76 (d, J = 8.8 Hz, 1H), 4.45 (s, 2H), 2.99 (t, J = 5.9 Hz, 2H), 2.53 (t, J = 7.0 Hz, 2H), 1.86 (m, 2H); 13 C NMR (151 MHz, DMSO-d6) δ 151.24, 142.56, 133.68, 131.96, 131.16, 130.25, 128.71, 127.95, 127.49, 120.78, 111.75, 56.93, 52.19, 30.61, 28.75.

[0132] Example 4: Preparation of compound T-4

[0133] Compound T-4 was prepared according to the synthetic procedure of Example 1. ESI-MS (m / z): 329.20 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) δ 11.16 (s, 1H), 8.99 (s, 1H), 7.74 - 7.64 (m, 2H), 7.43 (d, J = 8.1 Hz, 2H), 7.11 (d, J = 2.0 Hz, 1H), 6.92 - 6.78 (m, 2H), 4.41 (s, 2H), 3.20 - 3.09 (m, 2H), 2.89 - 2.80 (m, 2H), 2.15 (s, 3H), 1.84 (m, 2H); 13 C NMR (101 MHz, DMSO-d6) δ 164.59, 150.00, 143.26, 132.76, 131.81, 130.54, 129.82, 128.64, 128.10, 127.36, 119.61, 57.52, 53.08, 30.40, 29.67, 20.29.

[0134] Example 5: Preparation of compound T-5

[0135] Compound T-5 was prepared according to the synthetic procedure of Example 1. ESI-MS (m / z): 345.20 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) δ 7.92 - 7.86 (m, 2H), 7.50 (d, J = 8.2 Hz, 2H), 6.95 (d, J = 8.8 Hz, 1H), 6.90 (d, J = 3.0 Hz, 1H), 6.71 (dd, J = 8.8, 3.0 Hz, 1H), 4.39 (s, 2H), 3.67 (s, 3H), 3.04 (t, J = 5.4 Hz, 2H), 2.89 - 2.78 (m, 2H), 1.90 - 1.78 (m, 2H); 13 C NMR (101 MHz, DMSO-d6) δ 167.77, 154.33, 146.03, 145.10, 130.33, 130.09, 129.76, 128.36, 121.47, 117.34, 113.79, 58.10, 55.68, 53.49, 30.36, 29.69.

[0136] Example 6: Preparation of compound T-6

[0137] Compound T-6 was prepared according to the synthetic procedure of Example 1. ESI-MS (m / z): 333.20 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) δ 9.94 (d, J = 3.5 Hz, 1H), 7.85 - 7.68 (m, 2H), 7.53 (dt, J = 8.6, 1.0 Hz, 2H), 7.19 (dd, J = 8.1, 5.0 Hz, 1H), 7.01 (td, J = 8.2, 2.3 Hz, 1H), 6.46 (dd, J = 8.0, 2.2 Hz, 1H), 4.43 (t, J = 0.9 Hz, 2H), 3.46 (dt, J = 11.6, 5.9 Hz, 2H), 2.98 (ddd, J = 7.5, 6.6, 1.1 Hz, 2H), 2.06 - 1.97 (m, 2H); 13 C NMR (125 MHz, DMSO-d6) δ 167.79, 162.72, 1503.17, 140.56, 132.47, 130.83, 128.73, 127.94, 124.85, 109.76, 104.86, 56.82, 50.23, 36.75, 25.98.

[0138] Example 7: Preparation of compound T-7

[0139] Compound T-7 was prepared according to the synthetic procedure of Example 1. ESI-MS (m / z): 349.40 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) δ 9.94 (d, J = 3.5 Hz, 1H), 8.66 (d, J = 3.5 Hz, 1H), 7.85 - 7.79 (m, 2H), 7.43 (dt, J = 8.6, 1.0 Hz, 2H), 7.15 - 7.07 (m, 2H), 6.87 (d, J = 2.1 Hz, 1H), 4.61 (t, J = 1.0 Hz, 2H), 3.50 - 3.42 (m, 2H), 2.98 (t, J = 6.9 Hz, 2H), 2.01 (ddtd, J = 10.8, 7.1, 6.4, 5.6 Hz, 2H); 13 C NMR (125 MHz, DMSO-d6) δ 165.27, 150.29, 140.56, 134.62, 131.35, 130.59, 128.73, 127.94, 127.05, 122.84, 116.39, 56.82, 51.23, 33.04, 27.58.

[0140] Example 8: Preparation of compound T-8

[0141] Compound T-8 was prepared according to the synthetic procedure of Example 1. ESI-MS (m / z): 351.20 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) 1 H NMR (500 MHz, Chloroform-d) δ 9.97 (d, J = 3.4 Hz, 1H), 9.30 (d, J = 3.7 Hz, 1H), 7.71 - 7.64 (m, 2H), 7.51 (ddt, J = 9.4, 4.7, 1.0 Hz, 1H), 6.92 - 6.89 (m, 1H), 6.89 - 6.80 (m, 2H), 4.68 (d, J = 1.0 Hz, 2H), 3.50 - 3.41 (m, 2H), 3.00 (td, J = 7.0, 5.8 Hz, 2H), 2.02 (dtdd, J = 7.1, 6.4, 5.7, 4.8 Hz, 2H); 13C NMR (125 MHz, DMSO-d6) δ 165.36, 165.34, 160.18, 156.10, 146.72, 132.14, 131.26, 129.05, 128.76, 123.80, 117.81, 115.80, 114.73, 52.28, 51.23, 32.99, 27.58.

[0142] Example 9: Preparation of compound T-9

[0143] Compound T-9 was prepared according to the synthetic method of Example 1. ESI-MS (m / z): 368.20 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) δ 9.60 (d, J = 3.7 Hz, 1H), 9.29 (d, J = 3.5 Hz, 1H), 7.62 (dd, J = 8.0, 5.0 Hz, 1H), 7.35 (m, 1H), 6.92 - 6.89 (m, 1H), 6.89 - 6.80 (m, 2H), 4.65 (d, J = 1.0 Hz, 2H), 3.50 - 3.41 (m, 2H), 3.04 - 2.96 (m, 2H), 2.02 (m, 2H); 13 C NMR (125 MHz, DMSO-d6) δ 165.36, 165.34, 160.18, 156.10, 146.72, 132.14, 131.26, 129.05, 128.76, 123.80, 117.81, 115.80, 114.73, 52.28, 51.23, 32.99, 27.58.

[0144] Example 10: Preparation of compound T-10

[0145] T-10 was prepared based on the synthetic method of Example 1, and the synthetic route is as follows:

[0146] The specific synthesis operation is as follows:

[0147] 1) Refer to the synthetic method in Example 1 to prepare intermediate 6a.

[0148] 2) Intermediate ester 6a (5 mmol, 1 eq) was dissolved in 30 mL of chloroform, cooled to 0 °C, and then M-CPBA (5 mmol, 1 eq) was added and the reaction was stirred for about 30 minutes. TLC monitoring of the reaction was complete, and saturated sulfurous acid solution (about 10 mL) was slowly added to the above reaction solution and stirred for about 30 minutes, extracted with ethyl acetate, and purified by column chromatography (PE:EA = 10:1) to obtain the sulfoxide intermediate 7a. ESI-MS (m / z): 348.10 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) δ 7.93 - 7.86 (m, 2H), 7.54 - 7.47 (m, 2H), 7.33 (dd, J = 8.4, 4.4 Hz, 1H), 7.27 - 7.21 (m, 2H), 4.53 (d, J = 14.7 Hz, 1H), 4.32 (d, J = 14.7 Hz, 1H), 3.82 (s, 3H), 3.46 (ddd, J = 12.4, 8.6, 3.7 Hz, 1H), 3.03 (ddd, J = 12.3, 8.1, 4.0 Hz, 1H), 2.69 - 2.56 (m, 2H), 2.06 (m, 1H), 1.67 (m, 1H).

[0149] 3) Intermediate ester 7a (2 mmol, 1 eq) was added to 14 mL of freshly prepared hydroxylamine / KOH methanol solution (20 mmol, 10 eq), and the reaction was stirred in an ice bath for about 1-2 hours. TLC monitoring of the reaction was complete, and the reaction was adjusted to pH ≈ 6-7 using 1N HCl, and the reaction was precipitated as a solid, which was filtered and recrystallized with methanol / water to obtain the target compound T-10. ESI-MS (m / z): 349.15 [M+H] + ; 1 H NMR (600 MHz, DMSO-d6) δ 11.19 (s, 1H), 9.04 - 8.98 (m, 1H), 7.69 (d, J = 7.9 Hz, 2H), 7.42 (d, J = 8.0 Hz, 2H), 7.34 (dd, J = 8.7, 4.4 Hz, 1H), 7.25 (ddd, J = 19.2, 8.2, 3.0 Hz, 2H), 4.49 (d, J = 14.5 Hz, 1H), 4.28 (d, J = 14.6 Hz, 1H), 3.46 (ddd, J = 12.3, 8.5, 3.7 Hz, 1H), 3.04 (ddd, J = 12.3, 8.1, 4.0 Hz, 1H), 2.63 (ddd, J = 12.1, 8.1, 3.8 Hz, 1H), 2.57 (ddd, J = 12.5, 7.1, 3.2 Hz, 1H), 2.06 (m, 1H), 1.72 - 1.63 (m, 1H); 13C NMR (151 MHz, DMSO-d6) δ 164.52, 159.79, 143.07, 142.00, 141.26, 132.11, 128.49, 127.43, 122.83, 118.12, 111.65, 57.39, 52.78, 52.52, 25.38.

[0150] Example 11: Preparation of compound T-11

[0151] Compound T-11 was prepared according to the synthetic procedure of Example 10. ESI-MS (m / z): 365.10 [M+H] + ; 1 H NMR (600 MHz, DMSO-d6) δ 11.20 (s, 1H), 9.03 (s, 1H), 7.72 - 7.67 (m, 2H), 7.46 - 7.37 (m, 4H), 7.27 - 7.23 (m, 1H), 4.53 (s, 1H), 4.33 (d, J = 14.8 Hz, 1H), 3.41 (m, 1H), 3.16 - 3.08 (m, 1H), 2.79 - 2.66 (m, 2H), 2.08 (m, 1H), 1.76 (m, 1H); 13 C NMR (151 MHz, DMSO-d6) δ 170.82, 164.48, 146.26, 141.82, 140.01, 132.13, 131.22, 128.35, 127.48, 125.10, 122.30, 57.05, 52.91, 51.73, 25.29.

[0152] Example 12: Preparation of compound T-12

[0153] Compound T-12 was prepared according to the synthetic procedure of Example 10. ESI-MS (m / z): 410.90 [M+H] + ; 1H NMR (400 MHz, DMSO-d6) δ 7.63 (d, J = 7.9 Hz, 2H), 7.52 (d, J = 8.8 Hz, 2H), 7.38 (d, J = 7.8 Hz, 2H), 7.16 (d, J = 8.3 Hz, 1H), 4.49 (d, J = 14.7 Hz, 1H), 4.27 (d, J = 14.8 Hz, 1H), 3.38 (ddd, J = 12.4, 8.2, 3.8 Hz, 1H), 3.09 (ddd, J = 12.2, 7.6, 3.8 Hz, 1H), 2.77 - 2.59 (m, 2H), 2.14 - 1.99 (m, 1H), 1.80 - 1.64 (m, 1H); 13 C NMR (101 MHz, DMSO-d6) δ 164.53, 146.70, 141.81, 140.04, 134.07, 132.11, 128.33, 128.00, 127.45, 122.60, 115.93, 57.39, 52.87, 51.61, 25.66.

[0154] Example 13: Preparation of compound T-13

[0155] Compound T-13 was prepared according to the synthetic procedure of Example 10. ESI-MS (m / z): 445.40 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) δ 9.94 (d, J = 3.5 Hz, 1H), 8.66 (d, J = 3.5 Hz, 1H), 7.85 - 7.79 (m, 2H), 7.59 (d, J = 2.1 Hz, 1H), 7.43 (m, 2H), 7.37 - 7.31 (m, 1H), 6.89 (d, J = 8.0 Hz, 1H), 4.58 (t, J = 1.0 Hz, 2H), 3.55 (m, 2H), 2.97 - 2.89 (m, 2H), 2.15 - 2.03 (m, 2H); 13 C NMR (101 MHz, DMSO-d6) δ 165.27, 144.26, 140.56, 133.79, 131.46, 131.35, 129.90, 128.73, 128.04, 127.94, 117.76, 57.36, 54.82, 50.85, 21.19, 20.15.

[0156] Example 14: Preparation of compound T-14

[0157] Compound T-14 was prepared according to the synthetic procedure of Example 10. ESI-MS (m / z): 361.20 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) δ 7.66 (d, J = 8.2 Hz, 2H), 7.42 (d, J = 8.0 Hz, 2H), 7.24 (d, J = 8.7 Hz, 1H), 7.02 (d, J = 3.0 Hz, 1H), 6.96 (dd, J = 8.6, 3.0 Hz, 1H), 4.44 (d, J = 14.5 Hz, 1H), 4.24 (s, 1H), 3.45 (ddd, J = 12.5, 9.1, 3.6 Hz, 1H), 2.97 (ddd, J = 12.3, 8.4, 4.0 Hz, 1H), 2.58 - 2.50 (m, 2H), 2.03 (m, 1H), 1.61 (m, 1H); 13 C NMR (101 MHz, DMSO-d6) δ 165.36, 157.33, 142.27, 140.32, 139.19, 132.04, 128.47, 127.31, 122.18, 116.98, 109.18, 57.41, 55.90, 53.00, 52.86, 25.40.

[0158] Example 15: Preparation of compound T-15

[0159] Compound T-15 was prepared according to the synthetic procedure of Example 10. ESI-MS (m / z): 349.15 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) δ 9.94 (d, J = 3.5 Hz, 1H), 8.66 (d, J = 3.5 Hz, 1H), 7.85 - 7.78 (m, 3H), 7.43 (m, 2H), 7.09 (ddd, J = 9.0, 8.1, 2.3 Hz, 1H), 6.73 (dd, J = 8.0, 2.3 Hz, 1H), 4.62 (t, J = 0.9 Hz, 2H), 3.57 (m, 2H), 2.92 (m, 2H), 2.15 - 2.03 (m, 2H); 13 C NMR (101 MHz, DMSO-d6) δ 165.27, 165.16, 148.86, 140.56, 131.35, 131.28, 129.32, 128.73, 127.94, 110.71, 106.58, 57.30, 54.82, 50.80, 20.16.

[0160] Example 16: Preparation of compound T-16

[0161] Compound T-16 was prepared according to the synthetic procedure of Example 10. ESI-MS (m / z): 331.40 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) δ 9.94 (d, J = 3.5 Hz, 1H), 8.66 (d, J = 3.5 Hz, 1H), 7.85 - 7.79 (m, 2H), 7.66 (dd, J = 7.6, 1.5 Hz, 1H), 7.43 (m, 2H), 7.30 - 7.19 (m, 2H), 6.94 (dd, J = 6.9, 1.5 Hz, 1H), 4.58 (t, J = 1.0 Hz, 2H), 3.55 (ddd, J = 7.0, 6.2, 2.6 Hz, 2H), 2.95 - 2.87 (m, 2H), 2.08 (m, 2H); 13 C NMR (101 MHz, DMSO-d6) δ 165.27, 146.59, 140.56, 132.94, 131.89, 131.35, 128.73, 127.99, 127.94, 124.46, 118.51, 57.36, 54.87, 50.85, 20.15.

[0162] Example 17: Preparation of compound T-17

[0163] Compound T-17 was prepared according to the synthetic procedure of Example 10. ESI-MS (m / z): 367.20 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) δ 9.97 (d, J = 3.4 Hz, 1H), 9.30 (d, J = 3.7 Hz, 1H), 7.71 - 7.64 (m, 2H), 7.51 (m, 1H), 7.25 (dd, J = 8.1, 2.2 Hz, 1H), 7.05 (m, 1H), 7.01 (dd, J = 8.2, 5.1 Hz, 1H), 4.67 (d, J = 1.1 Hz, 2H), 3.57 (dd, J = 7.0, 6.2 Hz, 2H), 2.99 - 2.90 (m, 2H), 2.14 - 2.03 (m, 2H); 13 C NMR (101 MHz, DMSO-d6) δ 165.33, 160.18, 157.27, 143.84, 133.27, 132.14, 131.26, 128.76, 123.80, 120.75, 118.72, 115.31, 114.43, 54.78, 52.46, 50.80, 20.18.

[0164] Example 18: Preparation of compound T-18

[0165] T-18 was prepared based on the synthetic method of Example 1, and the synthetic route is as follows:

[0166] The specific synthesis operation is as follows:

[0167] 1) Refer to the synthetic method in Example 1 to prepare intermediate 6a.

[0168] 2) Intermediate ester 6a (5 mmol, 1 eq) was dissolved in 30 mL of chloroform, cooled to 0°C, then M-CPBA (15 mmol, 3 eq) was added and stirred for about 30 minutes. TLC monitoring of the complete reaction of raw materials, saturated sulfurous acid solution (about 10 mL) was slowly added to the above reaction liquid and stirred for about 30 minutes, extracted with ethyl acetate, purified by column chromatography (PE:EA=10:1) to obtain the sulfoxide intermediate 8a. ESI-MS (m / z): 365.10 [M+H] + ; 1 H NMR (600 MHz, DMSO-d6) δ 7.93-7.87 (m, 2H), 7.62-7.56 (m, 3H), 7.41 (ddd, J=9.0, 7.9, 3.2 Hz, 1H), 7.25 (dd, J=9.0, 4.5 Hz, 1H), 4.58 (s, 2H), 3.83 (s, 3H), 3.46 (dd, J=7.1, 4.8 Hz, 2H), 3.23-3.12 (m, 2H), 1.93 (m, 2H).

[0169] 3) Intermediate ester 8a (2 mmol, 1 eq) was added to 14 mL of freshly prepared hydroxylamine / KOH methanol solution (20 mmol, 10 eq), and the reaction liquid was stirred in an ice bath for about 1-2 hours. TLC monitoring of the complete reaction of raw materials, 1N HCl was used to adjust the pH of the reaction liquid to about 6-7, and the reaction liquid precipitated solid, which was filtered and recrystallized with methanol / water to obtain the target compound T-18. ESI-MS (m / z): 365.10 [M+H] + ; 1H NMR (400 MHz, DMSO-d6) δ 11.16 (s, 1H), 9.02 (s, 1H), 7.69 (d, J = 8.0 Hz, 2H), 7.57 (dd, J = 8.4, 3.1 Hz, 1H), 7.51 (d, J = 8.0 Hz, 2H), 7.40 (m, 1H), 7.25 (dd, J = 9.0, 4.5 Hz, 1H), 4.53 (s, 2H), 3.51 - 3.41 (m, 2H), 3.16 (t, J = 5.2 Hz, 2H), 1.93 (m, 2H); 13 C NMR (101 MHz, DMSO-d6) δ 164.45, 156.86, 145.95, 142.19, 135.17, 132.07, 128.46, 127.37, 123.08, 121.55, 114.20, 57.59, 53.81, 53.42, 24.80.

[0170] Example 19: Preparation of compound T-19

[0171] Compound T-19 was prepared according to the synthetic procedure of Example 18. ESI-MS (m / z): 381.40 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) δ 9.94 (d, J = 3.5 Hz, 1H), 8.66 (d, J = 3.5 Hz, 1H), 7.85 - 7.79 (m, 2H), 7.67 (d, J = 2.3 Hz, 1H), 7.43 (m, 2H), 7.33 (dd, J = 8.7, 2.3 Hz, 1H), 7.06 (d, J = 8.7 Hz, 1H), 4.63 (t, J = 1.0 Hz, 2H), 3.51 - 3.43 (m, 2H), 3.35 (m, 2H), 2.06 (m, 2H); 13 C NMR (101 MHz, DMSO-d6) δ 165.27, 144.96, 140.59, 133.56, 131.35, 128.73, 128.69, 128.63, 127.94, 127.52, 120.66, 56.40, 55.29, 50.69, 20.69.

[0172] Example 20: Preparation of compound T-20

[0173] Compound T-20 was prepared according to the synthetic procedure of Example 18. ESI-MS (m / z): 426.10 [M+H] + ; 1H NMR (400 MHz, DMSO-d6) δ 11.19 (s, 1H), 9.10 (s, 1H), 7.86 (d, J = 2.5 Hz, 1H), 7.75 - 7.67 (m, 2H), 7.63 (dd, J = 8.8, 2.5 Hz, 1H), 7.47 (d, J = 8.1 Hz, 2H), 7.10 (d, J = 9.0 Hz, 1H), 4.59 (s, 2H), 3.48 (d, J = 12.1 Hz, 2H), 3.37 - 3.22 (m, 2H), 1.96 (m, 2H); 13 C NMR (101 MHz, DMSO-d6) δ 164.34, 148.54, 141.80, 136.80, 134.66, 132.13, 129.92, 128.23, 127.45, 122.67, 112.23, 57.11, 53.64, 52.91, 24.64.

[0174] Example 21: Preparation of compound T-21

[0175] Compound T-21 was prepared according to the synthetic procedure of Example 18. ESI-MS (m / z): 361.20 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) δ 9.94 (d, J = 3.5 Hz, 1H), 8.66 (d, J = 3.5 Hz, 1H), 7.85 - 7.80 (m, 2H), 7.61 - 7.57 (m, 1H), 7.43 (dt, J = 8.5, 1.0 Hz, 2H), 7.26 - 7.20 (m, 1H), 7.02 (d, J = 8.4 Hz, 1H), 4.63 (t, J = 1.0 Hz, 2H), 3.51 - 3.43 (m, 2H), 3.35 (m, 2H), 2.06 (m, 2H); 13 C NMR (101 MHz, DMSO-d6) δ 165.27, 144.17, 140.59, 132.70, 131.93, 131.35, 130.18, 128.73, 128.41, 127.94, 117.67, 56.40, 55.30, 50.69, 20.80, 20.68.

[0176] Example 22: Preparation of compound T-22

[0177] Compound T-21 was prepared according to the synthetic procedure of Example 18. ESI-MS (m / z): 377.20 [M+H] + ; 1H NMR (400 MHz, DMSO-d6) δ 11.16 (s, 1H), 9.04 - 8.95 (m, 1H), 7.71 - 7.66 (m, 2H), 7.54 (d, J = 8.0 Hz, 2H), 7.33 (d, J = 3.0 Hz, 1H), 7.21 (d, J = 8.9 Hz, 1H), 7.13 (dd, J = 8.9, 3.1 Hz, 1H), 4.47 (s, 2H), 3.76 (s, 3H), 3.39 (dd, J = 7.6, 4.2 Hz, 2H), 3.05 (d, J = 6.1 Hz, 2H), 1.91 (m, 2H); 13 C NMR (101 MHz, DMSO-d6) δ 166.79, 165.27, 143.23, 140.41, 131.35, 130.38, 128.70, 127.94, 124.68, 109.90, 106.37, 59.82, 57.69, 52.64, 21.45.

[0178] Example 23: Preparation of compound T-23

[0179] Compound T-23 was prepared according to the synthetic procedure of Example 18. ESI-MS (m / z): 365.40 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) δ 9.94 (d, J = 3.5 Hz, 1H), 8.66 (d, J = 3.5 Hz, 1H), 7.86 - 7.79 (m, 3H), 7.43 (dt, J = 8.6, 1.0 Hz, 2H), 7.03 (ddd, J = 9.7, 7.9, 2.2 Hz, 1H), 6.84 (dd, J = 8.0, 2.3 Hz, 1H), 4.65 (t, J = 1.0 Hz, 2H), 3.49 (m, 2H), 3.33 (m, 2H), 2.06 (m, 2H); 13 C NMR (101 MHz, DMSO-d6) δ 166.79, 165.27, 143.23, 140.41, 131.35, 130.38, 128.70, 127.94, 124.68, 109.90, 106.37, 59.82, 57.69, 52.64, 21.45.

[0180] Example 24: Preparation of compound T-24

[0181] Compound T-24 was prepared according to the synthetic procedure of Example 18. ESI-MS (m / z): 383.40 [M+H] + ;1 H NMR (400 MHz, DMSO-d6) δ 9.97 (d, J = 3.4 Hz, 1H), 9.30 (d, J = 3.7 Hz, 1H), 7.71 - 7.64 (m, 2H), 7.51 (m, 1H), 7.45 (dd, J = 8.0, 2.0 Hz, 1H), 7.13 - 7.04 (m, 2H), 4.67 (d, J = 1.0 Hz, 2H), 3.49 (m, 2H), 3.41 - 3.31 (m, 2H), 2.06 (m, 2H); 13 C NMR (101 MHz, DMSO-d6) δ 165.33, 160.18, 156.89, 143.45, 132.14, 131.24, 128.70, 128.75, 123.80, 120.83, 120.14, 118.35, 114.43, 55.19, 52.13, 50.64, 20.70.

[0182] Example 25: Preparation of compound T-25

[0183] Compound T-25 was prepared according to the synthetic procedure of Example 18. ESI-MS (m / z): 347.40 [M+H] + ; 1 H NMR (400 MHz, DMSO-d6) δ 9.97 (d, J = 3.4 Hz, 1H), 9.30 (d, J = 3.7 Hz, 1H), 7.71 - 7.64 (m, 2H), 7.51 (m, 1H), 7.45 (dd, J = 8.0, 2.0 Hz, 1H), 7.13 - 7.04 (m, 2H), 4.67 (d, J = 1.0 Hz, 2H), 3.49 (m, 2H), 3.41 - 3.31 (m, 2H), 2.06 (m, 2H); 13 C NMR (101 MHz, DMSO-d6) δ 165.27, 146.99, 139.79, 133.56, 132.35, 129.83, 128.75, 128.40, 127.94, 123.82, 118.05, 59.40, 53.20, 49.97, 21.48.

[0184] Example 26: Inhibition activity of compounds against HDAC

[0185] The established experimental platform was used to test the conditions for detecting the inhibition effect of the compounds against HDAC6 and HDAC1 target sites, and SW-100 and Ricolinostat (ACY-1215) were used as positive control compounds.

[0186] Experimental materials:

[0187] The main experimental apparatus and reagents for in vitro HDAC6 and HDAC1 enzyme inhibition activity are as follows:

[0188] Experimental apparatus for in vitro HDAC enzyme activity test:

[0189] Table 1

[0190] Experimental reagents for in vitro HDAC enzyme activity test:

[0191] Table 2

[0192] Experimental method

[0193] Compound preparation:

[0194] Dissolve the sample in DMSO solution, prepare the appropriate stock concentration of the sample DMSO solution to be tested, and store the solution in the dark for standby.

[0195] HDAC1 / 6 enzymatic reaction process

[0196] 1) Preparation of reaction solution.

[0197] 2) Preparation of test compounds: The concentration of the test compound is set to 1 nM and 10 nM (HDAC1 test concentration is 100 nM and 500 nM), respectively, and each test concentration is tested in duplicate. Select appropriate final concentrations of SW-100 and Ricolinostat (ACY-1215), and dilute the solution to 8 concentration gradients with appropriate dilution multiples. The IC 50 value of the positive drug in a single well is detected as a control.

[0198] 3) Detection using BPS fluorescence-based HDAC activity assay in 384-well Source plates. The 10 mL HDAC reaction mixture used in the experiment is composed of HDAC analysis buffer, 100 mg BSA, different concentrations of test compounds, appropriate concentrations of HDACs, and 20 mM of fluorescent substrate.

[0199] 4) Incubate the reaction mixture at 37°C for 60 minutes, then add the developing solution containing trypsin and TSA to stop the reaction.

[0200] 5) After 20 minutes, use the enzyme marker to detect fluorescence at 360 nM and 460 nM.

[0201] Data analysis:

[0202] According to the read fluorescence signal value, calculate the percentage inhibition rate, and the calculation formula is as follows:

[0203] Wherein: Mean(Max) is the mean of the slope values of each Max well (with DMSO and enzyme); Mean(Min) is the mean of the slope values of each Min well (enzyme-free well); Sample Signal is the slope value of the compound well.

[0204] Fitting dose-effect curve: taking the log value of the compound concentration as the X axis, and the corresponding percentage inhibition rate as the Y axis, the dose-effect curve was fitted by using the log(inhibitor) vs. response-Variable slope of the analysis software GraphPad Prism 5, so as to obtain the IC 50 value of each compound for inhibiting enzyme activity.

[0205] The experimental results are shown in the following table:

[0206] Table 3 Inhibition rate of compounds at different concentrations on HDAC1 and HDAC6

[0207] Table 4 IC of compounds on HDAC1 and HDAC6 50

[0208] From the above table, it can be seen that the compounds of the present application show strong inhibitory activity (IC 50 <10nM) on HDAC6, and the inhibitory activity of some compounds is equivalent to or better than that of the positive control Rocilinostat (ACY-1215), SW-100. In addition, the inhibitory activity IC 50 of the compounds of the present application on HDAC1 is all >1000nM; relative to HDAC1, the compounds of the present application can selectively inhibit HDAC6, with a selectivity of >100 times, which is significantly better than that of Rocilinostat and SW-100.

[0209] Example 27: Protective activity of compounds on L-glutamic acid-induced damaged SH-SY5Y cells

[0210] The glutamic acid-induced neuronal damage and the possible protective effect of the test compounds were observed on the SH-SY5Y cell model, and compared with the positive test product.

[0211] Sample preparation: according to the recommended dissolution method, a mother liquor with a concentration of 1mM was prepared with physiological saline or DMSO, and then filtered and sterilized through a 0.22μm filter, and then diluted with the corresponding solvent in a certain proportion to obtain each dose of test product (i.e. the working solution concentration is 1, 10 and 100μM).

[0212] Negative control: sterile PBS, source: Wuhan Boster Bio-technical Co., Ltd.; batch number: S17B21; specification: 500 mL / bottle

[0213] Experimental instruments:

[0214] Table 5

[0215] Experimental reagents:

[0216] Table 6

[0217] Experimental method:

[0218] 1) SH-SY5Y neuron cell culture: Place the culture bottle containing cells in a cell culture incubator at 37°C, 5% CO2. Replace the fresh culture medium every 2-3 days, and pass the cells in time according to the cell density.

[0219] When passing, use 0.25% trypsin-EDTA solution to treat the cells for about 1 minute, and after the cells fall off from the bottom of the culture bottle, add culture medium to stop the action of the enzyme, then transfer the cell suspension to preheated fresh culture medium to stop the digestion. Next, collect the cells by centrifugation, and discard the supernatant. Resuspend the cell pellets in fresh culture medium, and make sure that the cells are fully dispersed by gently blowing or shaking to avoid the formation of cell clumps. Then, 5*10 4 Inoculate the cells into the 96-well plate, and let them stand for 2-4 hours to adhere.

[0220] 2) Glutamate treatment: First, determine the L-glutamate concentration according to the SH-SY5Y cell experiment.

[0221] The cultured cells are divided into normal culture group and model group. Remove the original culture medium, and add serum-free DMEM culture medium. Except for the normal group, the model group is treated with different concentrations of L-glutamate (10 / 25 / 50 / 100 / 200 / 500 μM) for 24 hours, and then subjected to MTT analysis to determine the cell survival rate. The L-glutamate concentration value at about 50% cell activity is taken as the glutamate concentration value for subsequent experiments.

[0222] Through the inhibition experiment of SH-SY5Y cells by different concentration gradients of glutamate, the glutamate concentration for subsequent modeling is determined to be 25 mM.

[0223] The cultured cells are divided into normal culture group, model group, and test drug group. Remove the original culture medium, and add serum-free DMEM culture medium. Except for the normal group and the model group, each drug group is first given pre-treatment with different concentrations of test substances for 0.5 hours, and then incubated with appropriate concentration of glutamate for 24 hours. After 24 hours, MTT analysis is performed to determine the cell survival rate.

[0224] 3) MTT method for measuring cell survival rate: specifically, 5 mg / ml MTT solution was added to each well to make the final concentration 0.5 mg / mL, and incubated in the incubator for 4 h, then the culture solution was discarded, 200 μL DMSO was added to each well, and the optical density OD value (measurement wavelength 570 nm) was read on the enzyme marker. The results were calculated according to the formula:

[0225] All data are expressed as mean ± standard error.

[0226] The selected compounds in the patent were tested for SH-SY5Y cell protection activity, and SW-100 and ACY-1215 were selected as controls, and the experimental results are as follows:

[0227] Table 7

[0228] The experimental results show that the compounds of the present application have certain protective effect on SH-SY5Y cells against L-glutamic acid-induced damage. In particular, compounds T-9, T-11, T-16, T-18, T-19, T-20, T-22 and T-24 have good in vitro protective activity, some of which are better than or equivalent to SW-100. It is suggested that the compounds of the present application have neuroprotective effect and can be used for the treatment of neurodegenerative related diseases.

[0229] Example 28: Test of in vitro anti-proliferative activity of compounds on tumor cells and human normal cells

[0230] The anti-proliferative activity of some compounds of the present application on brain glioma cell line U-87MG, myeloma cell line RPMI 8226, AtT-20 mouse pituitary tumor cells, human lung cancer cell line A549 and MRC-5 human normal embryonic lung fibroblast cells (cell source: ATCC) was determined under single concentration (10 μM) conditions, and Rocilinostat (ACY-1215) was selected as a control. The specific results are shown in the table (unit: Inh% in 10 μM):

[0231] Test method: take tumor cells in good growth state in logarithmic growth phase, inoculate 4×10 3 cells per well into a 96-well plate, set 8 replicate wells for each group, and place in a cell culture incubator for culture. After the cells adhere, transfect the corresponding plasmid, and continue to culture for 48 h. Discard the drug-containing culture medium, add 10 uL of freshly prepared CCK8 detection solution (brand: Engreen) to each well, and continue to culture in the incubator for 4 h. Then, measure the OD value at 450 nm using an enzyme marker. The experiment was repeated 3 times, and the average value of the experimental results was taken as the final experimental result, and the inhibition rate (%) was calculated.

[0232] Table 8

[0233] A: 100 > ~ > 90; B: 80 < ~ < 90; C: 70 < ~ < 80; D: 50 < ~ < 70 E: < 50

[0234] From the above table, it can be seen that, compared with the positive control ACY-1215, the compounds of the present application all showed good in vitro anti-tumor cell proliferation activity, and had high inhibitory activity on brain glioma cell line U-87MG, myeloma cell line RPMI8226 and AtT-20 mouse pituitary tumor cells. The anti-tumor cell proliferation activity of some compounds was better than that of the positive control drug.

[0235] At the same time, the compounds of the present application had weaker inhibitory activity on MRC-5 human normal embryonic lung fibroblasts than Rocilinostat, and had lower toxic side effects, which revealed that the compounds of the present application might have lower toxic side effects when used as anti-tumor drugs.

[0236] Example 29: Liver microsomal metabolic stability test

[0237] The rat and human liver microsomal stability of the compounds of the present application was determined, and the experimental operation was as follows:

[0238] 1. Experimental preparation

[0239] 1) Preheat 100 mM PB solution (0.1 M potassium phosphate buffer, 3 mM MgCl2, pH 7.4) at 37°C.

[0240] 2) Prepare liver microsomal solution (0.5 mg / mL):

[0241] Human liver microsomes 85 μL (20 mg / mL stock in 2963.1 μL PB)

[0242] Mouse liver microsomes 102 μL (16.4 mg / mL stock in 2946.1 μL PB)

[0243] 3) Prepare 20x NADPH (reduced coenzyme II) solution (20 mM NADPH):

[0244] NADPH, 3.0 mg, PB 180 μL

[0245] 4) Prepare 20x test compound and control compound solution (20 μM):

[0246] Compound (100 μM ethanol solution): 200 μL

[0247] PB: 800 μL

[0248] 2. Experimental procedure

[0249] Add SD rat or human liver microsomal solution to a 96-well plate, 180 μL per well. Add 10 μL of 20x NADPH solution to the liver microsomal solution and mix. Pre-warm the 20x test compound containing plate at 37°C for 10 min. Add 10 μL of 20x test compound solution to the 96-well plate and start the incubation. At 0, 5, 10, 20, 30, 60 min, take 20 μL of the above solution and add 100 μL of tolbutamide in acetonitrile to stop the reaction.

[0250] Centrifuge the stop plate at 5000xg (5000 times gravity acceleration) at 4°C for 15 min. Take 10 μL of supernatant to a 96-well plate pre-loaded with 190 μL of double distilled water, mix and analyze the sample using LC-MS / MS.

[0251] Test compound HPLC detection conditions:

[0252] Table 9

[0253] The mobile phase gradient is shown in the table below:

[0254] Table 10

[0255] The slope is measured by plotting the natural logarithm of the percentage of the remaining amount of the compound against time, and T is calculated according to the following formula: 1 / 2 and CL int where V / M is equal to 1 / protein concentration.

[0256] Table 11 Reaction half-life T of test compounds after incubation in rat and human liver microsomes 1 / 2 (min):

[0257] T-16* is the compound T-16 of patent 202010304883.0, the structure of which is as follows

[0258] As can be seen from the above table, the compound of the present application is more stable in metabolism in rat and human liver microsomes than the compound T-16 of patent 202010304883.0, and has better pharmacokinetic characteristics.

[0259] Example 30: hERG potassium channel affinity experiment

[0260] 1. Cell preparation

[0261] CHO-hERG cells were cultured in 175cm 2 When the cell density reached 60-80%, the culture solution was removed, and the cells were washed once with 7 mL PBS, and then 3 mL Detachin was added for digestion.

[0262] After complete digestion, 7 mL neutralizing solution was added, and then centrifuged, the supernatant was removed, and 5 mL culture solution was added for resuspension to ensure that the cell density was 2-5 x 10 6 / mL.

[0263] 2. Electrophysiological recording process

[0264] The single-cell high-impedance sealing and whole-cell mode forming process were all automatically completed by the Qpatch instrument. After obtaining the whole-cell recording mode, the cell was clamped at -80 mV. Before a 5-second +20 mV depolarization stimulus was given, a 50-millisecond -50 mV pre-voltage was first given, and then repolarized to -50 mV for 5 seconds, and then returned to -80 mV. This voltage stimulus was applied every 15 seconds, and recording was performed for 2 minutes. Then, extracellular recording was performed for 2 minutes, and then the drug administration process began. The compound concentration started from the lowest test concentration, and each test concentration was given for 2 minutes. After all the concentrations were continuously administered, the positive control compound 10 μM Cisapride was administered. At least 3 cells were tested for each concentration (n≥3).

[0265] 3. Compound preparation

[0266] The compound stock solution was diluted with extracellular solution, 2 μL of the compound stock solution was added to 998 μL of extracellular solution, and then 5-fold serial dilutions were performed in extracellular solution containing 0.2% DMSO to obtain the final concentration required for testing. Experimental data were analyzed by XLFit software.

[0267] The in vitro hERG potassium channel affinity activity of some compounds of the present application was preliminarily investigated, and the potential cardiac toxic side effects of the compounds were evaluated. The experimental results are as follows:

[0268] Table 12

[0269] The hERG experiment results showed that the half-inhibition concentration of the tested compounds of the present application on the hERG potassium ion channel was greater than 10 μM, indicating that the potential cardiac toxicity of the compounds of the present application was low.

[0270] Example 31: Mouse tail suspension experiment

[0271] 1. Animal grouping and drug administration amount

[0272] ICR male mice (SPF level), body weight about 16 grams. Animals were evenly grouped according to body weight, 10 in each sample group. ACY-1215 group and SW-100 group were set up, and the animals were transferred to the animal room for unified feeding after grouping.

[0273] 2. Test method

[0274] Each test group was injected intraperitoneally with the corresponding drug, with a drug volume of 0.1 ml / 10 g of body weight and a drug dose of 5 mg / kg. The mouse tail suspension test was performed 30 min after drug administration, with a duration of 6 min. The data were expressed as mean ± standard deviation (X ± SD, n = 10), and the results are shown in Figure 1, in which * represents P < 0.05 compared with the blank control group, and ** represents P < 0.01 compared with the blank control group. T-16* is the compound T-16 of patent 202010304883.0, and its structure is as follows

[0275] The experimental results show that: after the mice were given the samples for 30 min, the 5 mg / kg ACY1215 group and the SW-100 group did not reduce the immobility time of the mice, and there was no significant difference (P > 0.05); compared with the blank 1 group, the 5 mg / kg patent compounds T-1 and T-20 could reduce the immobility time of the mice by 55% and 48%, respectively, and there was a significant difference (P < 0.01); the 5 mg / kg patent 202010304883.0 compound T-16* could reduce the immobility time of the mice by 37%, and there was a significant difference (P < 0.05). It is suggested that the compounds of the present application show certain antidepressant activity, and the antidepressant activity is better than that of the patent 202010304883.0 compound T-16.

[0276] Example 32: Forced swimming test of rats

[0277] 1. Animal grouping and drug dosage

[0278] Wistar male rats (SPF level), body weight about 180 grams. Animals were evenly grouped according to body weight, and each sample group had one blank control group and five dose groups. A group of vortioxetine 20 mg / kg was set up, and the animals were transferred to the animal room for unified feeding after grouping.

[0279] 2. Test method

[0280] Each group of rats was given the corresponding drug by intraperitoneal administration, with a drug volume of 1 ml / 100 g of body weight and a drug dose of 20 mg / kg, continuously for one week.

[0281] One day before the last administration, the animals were fasted for 4 h, and then the rats were placed in a The rats were made to swim in a glass cylinder with water depth of 15 cm (water temperature of 25℃) for 15 min. One rat was put in each cylinder, and each rat was put in a clean water cylinder. After swimming, the rats were dried by a hot hair dryer.

[0282] The rat swimming test was performed 1 h after oral administration on the next day, and the swimming duration was 6 min. The time of swimming stop without movement within 6 min was recorded.

[0283] The data were expressed as mean ± standard deviation (X ± SD, n = 10), and the data difference was statistically analyzed by one-way analysis of variance (ANOVA), and the difference between groups was judged by P < 0.05. The experimental results are shown in FIG. 2, in which * represents P < 0.05 compared with the blank control group, and ** represents P < 0.01 compared with the blank control group.

[0284] T-16* is a compound T-16 of patent 202010304883.0, and the structure is as follows

[0285] The experimental results show that in the forced swimming test of rats, fluparoxantin and the compound T-20 of the present application can significantly reduce the swimming immobility time of rats at a dose of 20 mg / kg (fluparoxantin and T20 reduce the immobility time by 49% and 60%, respectively) (P < 0.01), and the compound T-1 of the present application can significantly reduce the swimming immobility time (by 35%) at the same dose (P < 0.05), showing better antidepressant activity. The antidepressant activity of the compounds T-1 and T-20 of the present application is better than that of the compound T-16 of patent 202010304883.0 (which reduces the immobility time by 17%) at the same dose, which is presumably related to more stable metabolism.

[0286] Although the specific embodiments of the present application are described above, those skilled in the art should understand that these are only illustrative, and various changes or modifications can be made to these embodiments without departing from the principles and essence of the present application. Therefore, the protection scope of the present application is defined by the appended claims.

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof: in, X1 is R 1a R 1b R 1c and R 1d Independently H, deuterium, halogen, cyano, hydroxyl or R 1 -L-; and when X1 is At that time, R 1a R 1b R 1c R 1d R 2a R 2b R 2c and R 2d They are not both hydrogen; -L- can be a linking bond, -(C1-C4 alkyl)-, -O-, -C(=O)- or -S(=O)2- independently; R 1 Independently C1-C6 alkyl, C3-C7 cycloalkyl, C6-C 10 Aryl, (R a R b N-, 3-7-membered heterocyclic alkyl or 5-10-membered heteroaryl, or, the aforementioned C1-C6 alkyl, C3-C7 cycloalkyl, C6-C 10 Aryl, (R a R b N-, 3-7-membered heterocyclic alkyl groups and 5-10-membered heteroaryl groups are each independently substituented by one or more R groups. c Substitution; the 3-7 membered heterocyclic alkyl group and R substituents of one or more groups. c In the substituted 3-7 membered heterocyclic alkyl group, the heteroatomic group is selected from one or more of N, O, S, S(=O), and S(=O)2, and the number of heteroatoms is 1-3; the 5-10 membered heteroaryl group and the one or more substituents R c In the substituted 5-10 heteroaryl group, the heterogroup is selected from one or more of N, O, and S, and the number of heteroatoms is 1-4; R c It can be independently hydroxyl, halogen, cyano, or C1-C4 alkyl; when there are multiple substituents, they can be the same or different; R a and R b Each is independently hydrogen or C1-C4 alkyl; R 2a R 2b R 2c and R 2d It is independently hydrogen, deuterium, halogen, C1-C4 alkyl, or C1-C4 alkyl substituted with one or more halogens; when there are multiple halogens, they may be the same or different.

2. The compound of formula (I) as claimed in claim 1, characterized in that, It meets one or more of the following conditions: (1)R 1a R 1b R 1c R 1d R 2a R 2b R 2c and R 2d In this context, the halogen is F, Cl, Br, or I; (2)R 1 In this context, the C1-C6 alkyl group is a C1-C3 alkyl group, such as methyl; (3)X1 is Preferred (4)R 1a For H; (5)R 1b and R 1c Independent of H, halogen or R 1 -L-; -L- is a hyphen or -O-, R 1 It is a C1-C6 alkyl group; preferably, R 1c It is H or halogen; (6)R 1d For H; (7)R 2a R 2b R 2c and R 2d It can be hydrogen or halogen independently.

3. The compound of formula (I) as described in claim 2, or a pharmaceutically acceptable salt thereof, characterized in that, It meets one or more of the following conditions: (1)R 1b and R 1c Independently halogen or R 1 -L; preferably, R 1b and R 1c Independently halogen or R 1 -L-,R 1a and R 1d For H; (2)R 1b It can be H, F, Cl, Br, methyl, or methoxy; (3)R 1c It can be H, F, or Cl; (4)R 2a R 2b R 2c and R 2d It can be hydrogen or F independently.

4. The compound of formula (I) as claimed in claim 1, or a pharmaceutically acceptable salt thereof, characterized in that, for In A and B, R 1b and R 1c Independently halogen, cyano, hydroxyl or R 1 -L-; -L- can be a linking bond, -(C1-C4 alkyl)-, -O-, -C(=O)- or -S(=O)2- independently; R 1 Independently C1-C6 alkyl, C3-C7 cycloalkyl, C6-C 10 Aryl, (R a R b N-, 3-7-membered heterocyclic alkyl or 5-10-membered heteroaryl, or, the aforementioned C1-C6 alkyl, C3-C7 cycloalkyl, C6-C 10 Aryl, (R a R b N-, 3-7-membered heterocyclic alkyl groups and 5-10-membered heteroaryl groups are each independently substituented by one or more R groups. c Substitution; the 3-7 membered heterocyclic alkyl group and R substituents of one or more groups. c In the substituted 3-7 membered heterocyclic alkyl group, the heteroatomic group is selected from one or more of N, O, S, S(=O), and S(=O)2, and the number of heteroatoms is 1-3; the 5-10 membered heteroaryl group and the one or more substituents R c In the substituted 5-10 heteroaryl group, the heterogroup is selected from one or more of N, O, and S, and the number of heteroatoms is 1-4; R c It can be independently hydroxyl, halogen, cyano, or C1-C4 alkyl; when there are multiple substituents, they can be the same or different; R a and R b Each is independently hydrogen or C1-C4 alkyl; Preferably, in A and B, R 1b and R 1c Independently halogen or R 1 -L-; -L- is used independently as a connector key or -O-; R 1 Independently, it is a C1-C6 alkyl group, preferably a C1-C3 alkyl group, such as methyl; Among C, D, E, and F, R 1b and R 1c The definition is as described in at least one of claims 1-3.

5. The compound of formula (I) as claimed in claim 1, characterized in that, -L- is used independently as a connector key or -O-; R 1 Independently, it is a C1-C6 alkyl group; R 2a R 2b R 2c and R 2d Independently hydrogen or halogen; When X1 is At that time, R 1a For hydrogen, R 1b and R 1c Independently halogen or R 1 -L-;R 1d H, halogen or R 1 -L-; When X1 is At that time, R 1a R 1b R 1c and R 1d Independent of H, halogen or R 1 -L-.

6. The compound of formula (I) as claimed in claim 1, characterized in that, The structure of the compound shown in formula (I) is as follows: Among them, R 2a and R 2d It is independently hydrogen or halogen; the halogen is preferably F; X1 and R 1b The definition is any of the following: Case 1: X1 is R 1b Halogen or R 1 -L-; -L- is -O-; R 1 Independently, it is a C1-C6 alkyl group, preferably a C1-C3 alkyl group, such as methyl; Case 2: X1 is R 1b H, halogen or R 1 -L-; -L- is -O-; R 1 Independently, it is a C1-C6 alkyl group, preferably a C1-C3 alkyl group, such as methyl; Preferably, the compound structure shown in formula (I) is as follows: Where X1 is X1 is preferred R 1b Halogen or R 1 -L-; -L- stands independently as -O-; R 1 Independently, it is a C1-C6 alkyl group, preferably a C1-C3 alkyl group, such as methyl; R 1b Halogens are preferred, such as F or Br.

7. The compound of formula (I) as claimed in claim 1, characterized in that, The compound represented by formula (I) has any of the following structures:

8. A method for preparing a compound as shown in formula (I), characterized in that, It includes the following steps: In a solvent, in the presence of a base, the compound of formula (II) is reacted with a hydroxylamine as shown below to give the compound of formula (I): Among them, X1, R 1a R 1b R 1c R 1d R 2a R 2b R 2c and R 2d The definition is as described in at least one of claims 1-7; R 3 It is H or C1-C6 alkyl; Preferably, the method for preparing the compound as shown in formula (I) satisfies one or more of the following conditions: (1) The solvent is an alcohol solvent, such as methanol; (2) The alkali is an alkali metal hydroxide, such as KOH; (3)R 3 It can be hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl; (4) The mass-to-volume ratio of the compound as shown in formula (II) to the solvent is 0.1 g / L to 20 g / L; (5) The molar ratio of the compound as shown in formula (II) to the hydroxylamine is 1:5 to 1:50, for example 1:10; (6) The temperature of the hydroxylation reaction is room temperature to 80°C; preferably 25-80°C; (7) The preparation method of the compound shown in formula (I) further includes post-treatment; the post-treatment includes one or more of the following steps: after the hydroxylation reaction is completed, adjust the pH to 6-7, filter to take the solid, recrystallize, and the product is ready; (8) The preparation method of the compound shown in formula (I) further includes the following step: in a solvent and in the presence of a base, the compound shown in formula (III) and the compound shown in formula (IV) are subjected to the following substitution reaction to obtain the compound shown in formula (II). Among them, X1, R 1a R 1b R 1c R 1d R 2a R 2b R 2c R 2d and R 3 The definition is as described in at least one of claims 1-7; X is a halogen, such as Br; Preferably, in the substitution reaction, the solvent is a nitrile solvent; Preferably, in the substitution reaction, the base is an alkali metal carbonate, such as K2CO3.

9. A compound as shown in formula (II) or a pharmaceutically acceptable salt thereof, in, X1, R 1a R 1b R 1c R 1d R 2a R 2b R 2c and R 2d The definition is as described in at least one of claims 1-7; R 3 It is H or C1-C6 alkyl.

10. The compound of formula (II) as claimed in claim 9, characterized in that, The compound represented by formula (II) has any of the following structures:

11. A pharmaceutical composition comprising a compound of formula (I) as described in at least one of claims 1-7, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers.

12. The use of a compound of formula (I) as described in at least one of claims 1-7, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described in claim 11 in the preparation of an HDAC inhibitor or a medicament; The HDAC inhibitor is preferably an HDAC6 inhibitor; The drug is preferably a drug for the prevention and / or treatment of cancer or nervous system diseases, or a drug for the prevention and / or treatment of diseases or conditions related to HDAC; the cancer is preferably pituitary adenoma, ovarian cancer, colon cancer, breast cancer, liver cancer, pancreatic cancer, gallbladder cancer, gastrointestinal cancer, head and neck cancer, cervical cancer, prostate cancer, lung cancer, melanoma, germ cell tumors, gestational trophoblastic tumors, glioblastoma, myeloma, neuroblastoma, CNS tumors, monocytic leukemia, B-cell leukemia, T-cell leukemia, B-cell lymphoma, T-cell lymphoma, or mast cell tumor; More preferably, it is glioblastoma, myeloma, pituitary adenoma, or lung cancer; the neurological disease is preferably stroke, depression, neurodegenerative disease, or peripheral neuropathy; the stroke is preferably ischemic stroke or hemorrhagic stroke; the depression is preferably major depressive disorder, persistent depressive disorder, bipolar disorder, seasonal affective disorder, or postpartum depression; the neurodegenerative disease is preferably Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, spinocerebellar degeneration, Cushing's syndrome, or Ritter syndrome; the peripheral neuropathy is preferably peroneal muscular atrophy or giant axonal neuropathy; The diseases or conditions associated with HDAC are preferably cancer or neurological diseases; The cancers are preferably pituitary adenoma, ovarian cancer, colon cancer, breast cancer, liver cancer, pancreatic cancer, gallbladder cancer, gastrointestinal cancer, head and neck cancer, cervical cancer, prostate cancer, lung cancer, melanoma, germ cell tumors, gestational trophoblastic tumors, glioblastoma, myeloma, neuroblastoma-derived CNS tumors, monocytic leukemia, B-cell leukemia, T-cell leukemia, B-cell lymphoma, T-cell lymphoma, or mast cell-derived tumors; more preferably glioblastoma, myeloma, pituitary adenoma, or lung cancer. The neurological disease is preferably stroke, depression, neurodegenerative disease or peripheral neuropathy; The stroke is preferably ischemic stroke or hemorrhagic stroke; The depression is preferably major depressive disorder, persistent depressive disorder, bipolar disorder, seasonal affective disorder, or postpartum depression; The neurodegenerative diseases mentioned are preferably Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, spinocerebellar degeneration, Cushing's syndrome, or Ritter syndrome; The peripheral neuropathy is preferably peroneal muscular atrophy or giant axial neuropathy.

13. The use of a compound V having the following structure in the preparation of a medicament; said medicament is a medicament for the prevention and / or treatment of depression; said depression is preferably major depressive disorder, persistent depressive disorder, bipolar disorder, seasonal affective disorder, or postpartum depression;

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