An HDAC6 inhibitor and its preparation method and application

Abstract

The present invention discloses a histone deacetylase 6 inhibitor, its preparation method, and use. The histone deacetylase 6 inhibitor comprises a monocyclic, fused, or spirocyclic nucleus and a hydrazide as a zinc ion chelating group. It has the structure shown in the following general formula (I). The present invention also provides a preparation method for such a compound and its use in pharmaceuticals for preventing or treating diseases associated with abnormal HDAC6 activity or expression. The compounds of the present invention exhibit significant inhibitory effects on HDAC6. #imgabs0#

Description

Technical Field

[0001] The present invention relates to the field of chemical synthesis, in particular to the field of organic compound synthesis and pharmaceutical application technology, and in particular to a hydrazide-containing histone deacetylase 6 inhibitor, a preparation method thereof, and an application thereof. Background Art

[0002] Epigenetic regulation plays an important role in cell and molecular regulation, among which histone acetylation and deacetylation are ubiquitous post-translational modification processes. Histone acetyltransferases (HATs) and histone deacetylases (HDACs) catalyze and maintain the balance of acetylation levels of lysine residues in nuclear and cytoplasmic proteins, which is crucial for cell homeostasis (Annu. Rev. Biochem. 2007, 76 (1), 75-100). It is worth noting that the imbalance of HDACs destroys this balance and leads to the occurrence of various diseases. HDAC is an enzyme family with multiple members. Currently, there are 18 subtypes known, which are divided into the following four categories according to their species and homology with yeast: Class I includes HDAC1, HDAC2, HDAC3, and HDAC8; Class IIa includes HDAC4, HDAC5, HDAC7, and HDAC9; Class IIb includes HDAC6 and HDAC10; Class III includes SIRT1 to SIRT7; Class IV, which has some homology with both Class I and Class II HDACs but has a different species, includes HDAC11. Among them, Class I, II, and IV are classic Zn 2+ Class III belongs to the Sirtuin family, which is NAD + Dependent HDACs. Among them, HDAC6 is mainly located in the cytoplasm and may regulate the functions of many disease-related proteins through its deacetylase pathway (Epigenomics, 2015, 7(1), 103–118). HDAC6 plays an important role in cell motility and invasiveness and is an attractive target for the treatment of various inflammatory diseases and neurodegenerative diseases. The study of selective HDAC6 inhibitors is of great value (Science, 2020, 369, 1448; Cells, 2021, 10(1), 12)

[0003] At present, the known histone deacetylase inhibitors can be divided into four categories according to their structures: 1. Hydroxamic acid analogs, with representative compounds including Vorinostat (approved for marketing in 2006), Belinostat (approved for marketing in 2014), and Panobinostat (approved for marketing in 2015); 2. Benzamide analogs, with representative compounds including Tucidinostat (approved for marketing in 2014), Entinostat (clinical phase III), and Mocetinostat (clinical phase II); 3. Cyclic peptides, with representative compounds including FK228 (Romidepsin, Istodax TM ) (approved for marketing in 2009); 4. Aliphatic carboxylic acids, representative compounds include Valproicacid (clinical phase III) and so on.

[0004] Most of the aforementioned inhibitors are non-selective HDAC inhibitors, inhibiting a wide range of signaling pathways and therefore exhibiting significant side effects. Currently known HDAC6 selective inhibitors, such as Rocilinostat (Phase III clinical trial), Tubastatin A, and Nexturastat A, mostly use hydroxamic acid as the zinc chelating group. However, hydroxamic acid HDAC6 inhibitors not only have poor selectivity for Class I HDACs but also exhibit poor pharmacokinetic properties and toxic metabolites in vivo. These limitations limit the clinical application of hydroxamic acid selective HDAC6 inhibitors. Therefore, developing more active and selective HDAC6 inhibitors is an urgent need. Summary of the Invention

[0005] The first objective of the present invention is to provide a compound that is an HDAC6 inhibitor with a monocyclic, fused, or spirocyclic ring as the parent nucleus and a hydrazide as the zinc ion chelating group. Based on references, the present invention designed and synthesized this compound for selective HDAC6 inhibition. In vitro anti-inflammatory activity tests demonstrated that the compound exhibited good anti-inflammatory activity and excellent HDAC6 inhibition.

[0006] The second purpose of the present invention is to provide a method for preparing the compound, which is rationally designed and the synthesized compound is stable.

[0007] The third object of the present invention is to provide the use of the above compound.

[0008] To develop potential HDAC6 selective inhibitors, we thoroughly investigated the structural characteristics of current HDAC6 inhibitors and the unique properties of the HDAC6 protein, analyzing their shortcomings. We incorporated a hydrazide structure, widely used in class I HDACs inhibitors, into the structure of HDAC6 inhibitors. Based on the HDAC6 protein structure, we conducted molecular design and successfully validated a novel HDAC6 selective inhibitor.

[0009] Specifically, the present invention provides a compound containing a monocyclic, fused or spirocyclic ring as a parent core and hydrazide as a zinc ion chelating group.

[0010] 1. The present invention provides a compound of formula (I) or a pharmaceutically acceptable salt thereof, characterized in that its structure is as follows:

[0011]

[0012] in

[0013] Z is selected from a bond, (C1-C 12 )alkylene, (C2-C 12 )alkenylene, (C2-C 12 )alkynylene, heterocyclic or (C1-C 12 )alkylamide, wherein Z is preferably selected from a bond, a (C1-C4)alkyl, a 5-membered heterocycle or

[0014] Y is selected from H, (C1-C 12 )alkyl, (C2-C 12 )alkenyl, (C2-C 12 ) alkynyl, halogen, Among them, Y is preferably selected from H, (C3-C5) alkynyl, halogen,

[0015] R6 is selected from H, optionally substituted (C1-C 12 )alkyl, wherein R6 is preferably selected from H, optionally substituted (C1-C4)alkyl;

[0016] R is selected from a 5-14 membered aliphatic ring, aliphatic heterocyclic ring, aromatic ring, heteroaromatic ring, condensed aromatic hydrocarbon, benzene condensed heterocyclic ring or condensed heterocyclic ring,

[0017] R is optionally substituted by one or more substituents, preferably substituted by 1-3 substituents, wherein the substituents are selected from halogen, haloalkyl, alkyl, alkenyl, alkynyl, alkoxy, hydroxyl, phenolic hydroxyl, cyano or formate, preferably the substituents are selected from halogen, halo (C1-C 12 )alkyl, (C1-C12 )alkyl, (C2-C 12 )alkenyl, (C3-C5)alkynyl, (C1-C2)alkoxy, hydroxy, phenolic hydroxy, cyano or (C1-C2)formate;

[0018] R1 is selected from H, (C1-C 12 )alkyl, R1 is preferably selected from H, (C1-C2)alkyl;

[0019] W is selected from C or N.

[0020] 2. The compound according to item 1 or a pharmaceutically acceptable salt thereof, wherein R is

[0021]

[0022] Ring A is selected from a 5-6 membered aliphatic ring, aliphatic heterocyclic ring, aromatic ring or heteroaromatic ring,

[0023] Ring A is optionally substituted by one or more substituents selected from halogen, haloalkyl, alkyl, alkenyl, alkynyl, alkoxy, phenolic hydroxyl, cyano or formate, preferably selected from halogen, halo (C1-C 12 )alkyl, (C1-C 12 )alkyl, (C2-C 12 )alkenyl, (C3-C5)alkynyl, (C1-C2)alkoxy, phenolic hydroxyl, cyano or (C1-C2)formate.

[0024] 3. The compound according to item 2 or a pharmaceutically acceptable salt thereof, wherein ring A is selected from one or more of the following ring systems:

[0025] 4. The compound according to item 3 or a pharmaceutically acceptable salt thereof, wherein R is

[0026] Wherein, the B ring is selected from a 5-6 membered aliphatic ring, aliphatic heterocyclic ring, aromatic ring or heteroaromatic ring,

[0027] Ring B is optionally substituted by one or more substituents selected from halogen, haloalkyl, alkyl, alkenyl, alkynyl, alkoxy, phenolic hydroxyl, cyano or formate, preferably selected from halogen, halo (C1-C 12 )alkyl, (C1-C 12 )alkyl, (C2-C 12 )alkenyl, (C3-C5)alkynyl, (C1-C2)alkoxy, phenolic hydroxyl, cyano or (C1-C2)formate,

[0028] X is selected from CH2, O, S, (C=O),

[0029] 5. The compound according to item 4 or a pharmaceutically acceptable salt thereof, wherein Ring B is selected from one or more of the following ring systems:

[0030] 6. The compound according to item 1 or a pharmaceutically acceptable salt thereof, wherein R is

[0031] in,

[0032] Ring C is selected from H, 5-6 membered aromatic ring or aromatic heterocyclic ring,

[0033] The C ring is optionally substituted by one or more substituents selected from halogen, haloalkyl, alkyl, alkenyl, alkynyl, alkoxy, phenolic hydroxyl, cyano or formate, and the substituents are preferably selected from halogen, halo (C1-C 12 )alkyl, (C1-C 12 )alkyl, (C2-C 12 )alkenyl, (C3-C5)alkynyl, (C1-C2)alkoxy, phenolic hydroxyl, cyano or (C1-C2)formate.

[0034] 7. The compound according to item 6 or a pharmaceutically acceptable salt thereof, wherein the C ring is selected from one or more of the following ring systems: The C ring is preferably pyridine.

[0035] 8. The compound according to any one of items 1 to 7, or a pharmaceutically acceptable salt thereof, which is:

[0036] 4-((9H-carbazol-9-yl)methyl)-N'-ethylbenzohydrazide;

[0037] N'-ethyl-4-((1,2,3,4-tetrahydro-9H-carbazol-9-yl)methyl)benzohydrazide;

[0038] 4-((9H-pyrido[2,3-b]indol-9-yl)methyl)-N'-ethylbenzohydrazide;

[0039] 4-((9H-pyrido[3,4-b]indol-9-yl)methyl)-N'-ethylbenzohydrazide;

[0040] 4-((5H-pyrido[4,3-b]indol-5-yl)methyl)-N'-ethylbenzohydrazide;

[0041] 4-(Acridin-10(9H)-ylmethyl)-N'-ethylbenzohydrazide;

[0042] 4-((10H-Benzoxazin-10-yl)methyl)-N'-ethylbenzohydrazide;

[0043] 4-((10H-phenothiazin-10-yl)methyl)-N'-ethylbenzohydrazide;

[0044] 4-((10H-benzo[b]pyrido[2,3-e][1,4]thiazin-10-yl)methyl)-N'-ethylbenzohydrazide;

[0045] 4-((10H-benzo[b]pyrido[2,3-e][1,4]thiazin-10-yl)methyl)-N'-methylbenzohydrazide;

[0046] 4-((10,11-dihydro-5H-dibenzo[b,f]azepin-5-yl)methyl)-N'-ethylbenzohydrazide;

[0047] 4-((5H-dibenzo[b,f]azepin-5-yl)methyl)-N'-ethylbenzohydrazide;

[0048] 4-((11H-benzo[b]pyrido[4,3-f]heptidin-11yl)methyl)-N'-ethylbenzohydrazide;

[0049] N'-ethyl-4-((10-oxo-10,11-dihydro-5H-dibenzo[b,f]heptidin-5-yl)methyl)benzohydrazide;

[0050] 4-(((2,4-Dimethyl-5H-benzo[b]pyrimidin[4,5-f]azepin-5-yl]methyl)methyl)-N'-ethylbenzohydrazide;

[0051] N'-ethyl-4-((3-(trifluoromethyl)-5H-dibenzo[b,f]azepin-5-yl)methyl)benzohydrazide;

[0052] N'-ethyl-4-((10-hydroxy-10,11-dihydro-5H-dibenzo[b,f]azepin-5-yl)methyl)benzohydrazide;

[0053] N'-ethyl-4-((3-methyl-5H-dibenzo[b,f]azepin-5-yl)methyl)benzohydrazide;

[0054] N'-ethyl-4-((3-fluoro-5H-dibenzo[b,f]azepin-5-yl)methyl)benzohydrazide;

[0055] N-(4-(2-ethylhydrazine-1-carbonyl)benzyl)benzamide;

[0056] 1-Butyl-1-(4-(2-ethylhydrazine-1-carbonyl)benzyl)-3-phenylurea;

[0057] N-Butyl-N-(4-(2-ethylhydrazine-1-carbonyl)benzyl)benzamide;

[0058] 1-(4-(2-ethylhydrazine-1-carbonyl)phenyl)-3-(quinolin-8-yl)urea;

[0059] 1-(4-(2-ethylhydrazine-1-carbonyl)benzyl)-3-(quinolin-8-yl)urea;

[0060] N'-ethyl-4-((5-(thiophen-2-yl)-1H-tetrazol-1-yl)methyl)benzohydrazide;

[0061] N'-ethyl-2-(pyridin-3-yl)thiazole-4-carbohydrazide;

[0062] N'-ethyl-4-((quinolin-8-ylamino)methyl)benzohydrazide;

[0063] 4-((2,4-dioxy-3-phenethyl-3,4-dihydroquinazolin-1(2H)-yl)methyl)-N'-ethylbenzohydrazide;

[0064] N'-ethyl-4-((5-(4-methoxybenzoyl)-1H-indol-1-yl)methyl)benzohydrazide;

[0065] 4-((4',5'-dihydrospiro[piperidin-4,7'-thieno[2,3-c]pyran]-1-yl)methyl)-N'-ethylbenzohydrazide;

[0066] 2-(4-bromophenyl)-N'-ethyloxazole-4-carbohydrazide;

[0067] 8-(2-ethylhydrazinyl)-8-oxo-N-phenyloctanamide;

[0068] N-(4-(2-methylhydrazine-1-carbonyl)benzyl)benzamide; or

[0069] 8-(2-Methylhydrazinyl)-8-oxo-N-phenyloctanamide.

[0070] 9. A method for preparing the compound or a pharmaceutically acceptable salt thereof according to any one of items 1 to 8, comprising the following steps:

[0071]

[0072] 10. The preparation method according to item 9, wherein

[0073] The intermediate 1 can be obtained by the following method: using a polycyclic compound containing an aromatic secondary amine as a raw material, reacting it with methyl 4-(bromomethyl)benzoate under the action of NaH to obtain the intermediate 1.

[0074] 11. The preparation method according to item 9, wherein

[0075] The intermediate 1 can be obtained as follows: a compound containing an aromatic amino group is used as a raw material, reacted with triphosgene under alkaline conditions to generate an isocyanate intermediate, and then reacted with substituted or unsubstituted methyl 4-aminomethylbenzoate hydrochloride or methyl 4-aminobenzoate to obtain intermediate 1.

[0076] 12. The preparation method according to item 9, wherein

[0077] The intermediate 1 can be obtained by the following method: using an aryl formic acid compound as a raw material, condensing it with substituted or unsubstituted 4-aminomethylbenzoic acid methyl ester hydrochloride or 4-aminobenzoic acid methyl ester under the action of HATU to obtain the intermediate 1.

[0078] 13. A pharmaceutical composition comprising the compound according to any one of items 1 to 8 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable diluent or carrier.

[0079] 14. A compound according to any one of items 1 to 8, or a pharmaceutically acceptable salt thereof, for use as a medicament.

[0080] 15. Use of the compound according to any one of items 1 to 8 or a pharmaceutically acceptable salt thereof in the preparation of an HDAC6 inhibitor.

[0081] 16. Use of the compound according to any one of items 1 to 8 or a pharmaceutically acceptable salt thereof in the preparation of a treatment for a disease associated with abnormal expression of HDAC6 activity.

[0082] 17. The use according to item 16, wherein the disease associated with abnormal expression of HDAC6 activity includes tumors, neurodegenerative diseases, nervous system diseases, stroke, inflammation or autoimmune diseases.

[0083] 18. The method of claim 16, wherein the tumor comprises triple-negative breast cancer, lung cancer, melanoma, esophageal cancer, prostate cancer, breast cancer, cervical cancer, ovarian cancer, gastric cancer, pancreatic cancer, bladder cancer, colorectal cancer, brain tumor, glioma, anaplastic oligodendroglioma, adult glioblastoma, adult anaplastic astrocytoma, bone cancer, and soft tissue sarcoma.

[0084] 19. The method of claim 16, wherein the neurodegenerative disease comprises Parkinson's disease (PD), Alzheimer's disease (AD), cerebral ischemia (CI), brain injury (BI), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS), different types of spinocerebellar ataxia (SCA), epilepsy, Huntington's chorea, and Pick's disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0085] Figure 1 Example 13 Selective inhibition of HDAC6 in two cell lines

[0086] Figure 2 Figure 3. Changes of HDAC6 inhibitory activity in some examples with enzyme and compound incubation time.

[0087] Figure 3 Some examples and test diagrams of HDAC6 enzyme dissociation ability

[0088] Figure 4 Example 13 and IL-1β level-relationship diagram DETAILED DESCRIPTION

[0089] Compounds of the present invention

[0090] A compound of formula (I) as shown below or a pharmaceutically acceptable salt thereof:

[0091]

[0092] in

[0093] Z is selected from a bond, (C1-C 12 )alkylene, (C2-C 12 )alkenylene, (C2-C 12 )alkynylene, heterocyclic or (C1-C 12 )alkylamide, wherein Z is preferably selected from a bond, a (C1-C4)alkyl, a 5-membered heterocycle or

[0094] Y is selected from H, (C1-C 12 )alkyl, (C2-C 12 )alkenyl, (C2-C 12 ) alkynyl, halogen, Among them, Y is preferably selected from H, (C3-C5) alkynyl, halogen,

[0095] R6 is selected from H, optionally substituted (C1-C 12 )alkyl, wherein R6 is preferably selected from H, optionally substituted (C1-C4)alkyl;

[0096] R is selected from a 5-14 membered aliphatic ring, aliphatic heterocyclic ring, aromatic ring, heteroaromatic ring, condensed aromatic hydrocarbon, benzene condensed heterocyclic ring or condensed heterocyclic ring,

[0097] R is optionally substituted by one or more substituents, preferably substituted by 1-3 substituents, selected from halogen, haloalkyl, alkyl, alkenyl, alkynyl, alkoxy, hydroxy, phenolic hydroxy, cyano or formate, preferably selected from halogen, halo(C1-C 12 )alkyl, (C1-C 12 )alkyl, (C2-C 12 )alkenyl, (C3-C5)alkynyl, (C1-C2)alkoxy, hydroxy, phenolic hydroxy, cyano or (C1-C2)formate;

[0098] R1 is selected from H, (C1-C 12 )alkyl, R1 is preferably selected from H, (C1-C2)alkyl;

[0099] W is selected from C or N.

[0100] In certain preferred embodiments of the present invention, certain groups in the compound shown in I are defined as follows (undefined groups are the same as those described in any scheme of this application).

[0101] In some embodiments, Z is a bond; in some embodiments, Z is a 5-membered heterocyclic ring containing 1 heteroatom selected from N, O, and S. In some embodiments, Z is a 5-membered heterocyclic ring containing 2 heteroatoms selected from N, O, and S. In some embodiments, Z is

[0102] In some embodiments, Z is a bond, and the compound of formula (I) or a pharmaceutically acceptable salt thereof has the following structure:

[0103]

[0104] In some embodiments, Z is a 5-membered ring, for example, a 5-membered heterocyclic ring containing one heteroatom, and the compound of formula (I) or a pharmaceutically acceptable salt thereof may have the following structure:

[0105]

[0106] In some embodiments, Z is a 5-membered ring, for example, a 5-membered heterocyclic ring containing two heteroatoms, and the compound of formula (I) or a pharmaceutically acceptable salt thereof may have the following structure:

[0107]

[0108] In some embodiments, Z is The compound of formula (I) or a pharmaceutically acceptable salt thereof has the following structure:

[0109]

[0110] In some embodiments, Y is H; in some embodiments, Y is selected from (C3-C5) alkynyl; in some embodiments, Y is selected from one of the halogens, such as F, Cl, Br, I; in some embodiments, Y is selected from

[0111] In some embodiments, Y is H, and the compound of formula (I) or a pharmaceutically acceptable salt thereof has the following structure:

[0112]

[0113] In some embodiments, when Y is H, Z is When the compound of formula (I) or a pharmaceutically acceptable salt thereof has the following structure:

[0114]

[0115] In some embodiments, Y is a halogen, for example, Y is Br, and the compound of formula (I) or a pharmaceutically acceptable salt thereof has the following structure:

[0116]

[0117] In some embodiments, when Y is a halogen, for example, Y is Br, and Z is a 5-membered ring, for example, a 5-membered heterocycle, the compound of formula (I) or a pharmaceutically acceptable salt thereof may have the following structure:

[0118]

[0119] In some embodiments, Y is The compound of formula (I) or a pharmaceutically acceptable salt thereof has the following structure:

[0120]

[0121] In some embodiments, when Y is When Z is a bond, the compound of formula (I) or a pharmaceutically acceptable salt thereof has the following structure:

[0122]

[0123] In some embodiments, Y is The compound of formula (I) or a pharmaceutically acceptable salt thereof has the following structure:

[0124]

[0125] In some embodiments, when Y is When Z is a bond, the compound of formula (I) or a pharmaceutically acceptable salt thereof has the following structure:

[0126]

[0127] In some embodiments, R6 is H; in some embodiments, R6 is selected from (C1-C4) alkyl, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl.

[0128] In some embodiments, R is selected from a 5-14 membered aliphatic ring, an aliphatic heterocyclic ring, an aromatic ring or a heteroaromatic ring, a fused aromatic hydrocarbon, a benzene fused heterocyclic ring, or a fused heterocyclic ring, optionally substituted with one or more substituents, wherein the heteroatoms are selected from N, O, and S. In some embodiments, R is selected from a 5-12 membered ring system, wherein the heteroatoms are selected from N, O, and S. In some embodiments, R is selected from a 5-10 membered ring system, wherein the heteroatoms are selected from N, O, and S. In some embodiments, R is selected from a 5-9 membered ring system, wherein the heteroatoms are selected from N, O, and S. In some embodiments, R is selected from a 5-8 membered ring system, wherein the heteroatoms are selected from N, O, and S. In some embodiments, R is selected from a 5-7 membered ring system, wherein the heteroatoms are selected from N, O, and S. In some embodiments, R is selected from a 5-6 membered ring system, wherein the heteroatoms are selected from N, O, and S.

[0129] In some embodiments, the ring system selected from R is 5-membered. In some embodiments, the ring system selected from R is 6-membered. In some embodiments, the ring system selected from R is 7-membered. In some embodiments, the ring system selected from R is 8-membered. In some embodiments, the ring system selected from R is 9-membered. In some embodiments, the ring system selected from R is 10-membered. In some embodiments, R 1 In some embodiments, the ring system selected from R is 11-membered. In some embodiments, the ring system selected from R is 12-membered. In some embodiments, the ring system selected from R is 13-membered. In some embodiments, the ring system selected from R is 14-membered. In some embodiments, the heteroatom is N. In some embodiments, the heteroatom is O. In some embodiments, the heteroatom is S. In some embodiments, the heteroatom is N and O. In some embodiments, the heteroatom is N and S. In some embodiments, the heteroatom is O and S.

[0130] In some embodiments, examples of R include, but are not limited to, oxirane, thiol, aziridine, oxetane, thietane, azetidine, tetrahydrofuran, tetrahydrothiophene, pyrrolidine, tetrahydropyran, tetrahydrothiopyran, piperidine, 1,4-dioxane, 1,4-oxathiinane, morpholine, 1,4-dithiane, piperazine, 1,4-azathiane, azepane, thiepane, azepane, 1,4-dioxane, 1,4-oxathiinane, 1,4-azaoxane, 1,4-dithiinane, 1,4-diazepane, tropane, quinuclidine, dihydropyran, dihydropyridine, tetrahydropyridine, and pyrrolidone, optionally substituted with one or more substituents.

[0131] In some embodiments, R is substituted with one substituent; in some embodiments, R is substituted with two substituents; in some embodiments, R is substituted with three substituents.

[0132] In some embodiments, the substituent is one or more halogens, such as F, Cl, Br, I; in some embodiments, the halogen atom is F; in some embodiments, the halogen atom is Cl; in some embodiments, the halogen atom is Br. In some embodiments, the halogen atom is I; in the following embodiments, the substituent is a haloalkyl group, wherein the halogen atom is selected from F, Cl, Br, I; in some embodiments, the substituent is (C1-C 12 )alkyl, (C2-C 12 )alkenyl, (C3-C5)alkynyl, (C1-C2)alkoxy, phenolic hydroxyl, cyano or (C1-C2)formate.

[0133] In some embodiments, R is a benzene ring, when Y is When the compound of formula (I) or a pharmaceutically acceptable salt thereof has the following structure:

[0134]

[0135] In some embodiments, R is a benzene ring, when Y is When the compound of formula (I) or a pharmaceutically acceptable salt thereof has the following structure:

[0136]

[0137] In some embodiments, R is a benzene ring, when Y is When the compound of formula (I) or a pharmaceutically acceptable salt thereof has the following structure:

[0138]

[0139] In some embodiments, R is quinoline, when Y is When the compound of formula (I) or a pharmaceutically acceptable salt thereof has the following structure:

[0140]

[0141] In some embodiments, R is quinoline, when Y is When the compound of formula (I) or a pharmaceutically acceptable salt thereof has the following structure:

[0142]

[0143] In some embodiments, R is quinoline, when Y is When the compound of formula (I) or a pharmaceutically acceptable salt thereof has the following structure:

[0144]

[0145] In some embodiments, R is a fused heterocyclic ring, which may contain one heteroatom. For example, R is carbazole, when Y is When the compound of formula (I) or a pharmaceutically acceptable salt thereof has the following structure:

[0146]

[0147] In some embodiments, R is a fused heterocyclic ring, which may contain one heteroatom. For example, R is When Y is When the compound of formula (I) or a pharmaceutically acceptable salt thereof has the following structure:

[0148]

[0149] In some embodiments, R is a fused heterocyclic ring, which may contain one heteroatom. For example, R is When Y is When the compound of formula (I) or a pharmaceutically acceptable salt thereof has the following structure:

[0150]

[0151] In some embodiments, R is a fused heterocyclic ring, which may contain one heteroatom. For example, R is When Y is When the compound of formula (I) or a pharmaceutically acceptable salt thereof has the following structure:

[0152]

[0153] In some embodiments, R is a fused heterocyclic ring, which may contain two heteroatoms. For example, R is When Y is When the compound of formula (I) or a pharmaceutically acceptable salt thereof has the following structure:

[0154]

[0155] In some embodiments, R is a fused heterocyclic ring, which may contain two heteroatoms. For example, R is When Y is When the compound of formula (I) or a pharmaceutically acceptable salt thereof has the following structure:

[0156]

[0157] In some embodiments, R is a fused heterocyclic ring, which may contain two heteroatoms. For example, R is When Y is When the compound of formula (I) or a pharmaceutically acceptable salt thereof has the following structure:

[0158]

[0159] In some embodiments, R is a fused heterocyclic ring, which may contain three heteroatoms. For example, R is When Y is When the compound of formula (I) or a pharmaceutically acceptable salt thereof has the following structure:

[0160]

[0161] In some embodiments, R is a fused heterocyclic ring, which may contain three heteroatoms. For example, R is When Y is When the compound of formula (I) or a pharmaceutically acceptable salt thereof has the following structure:

[0162]

[0163] In some embodiments, R is a fused heterocycle, optionally substituted by one or more substituents, for example, the substituent is halogen, and R can be When Y is When the compound of formula (I) or a pharmaceutically acceptable salt thereof has the following structure:

[0164]

[0165] In some embodiments, R is a fused heterocyclic ring, optionally substituted by one or more substituents, for example, the substituent is a haloalkyl group, and R can be When Y is When the compound of formula (I) or a pharmaceutically acceptable salt thereof has the following structure:

[0166]

[0167] In some embodiments, R is a fused heterocyclic ring, optionally substituted by one or more substituents, for example, if the substituent is hydroxyl, R can be When Y is When the compound of formula (I) or a pharmaceutically acceptable salt thereof has the following structure:

[0168]

[0169] In some embodiments, R1 is H; in some embodiments, R1 is (C1-C2) alkyl, such as methyl or ethyl.

[0170] In some embodiments, W is C, and the compound of formula (I) or a pharmaceutically acceptable salt thereof has the following structure:

[0171]

[0172] In some embodiments, W is N, and the compound of formula (I) or a pharmaceutically acceptable salt thereof has the following structure:

[0173]

[0174] In some embodiments, W is N, Z is a 5-membered ring, for example, a 5-membered heterocycle containing two heteroatoms, and the compound of formula (I) or a pharmaceutically acceptable salt thereof may have the following structure:

[0175]

[0176] In certain preferred embodiments of the present invention, certain groups in the compound shown in I are defined as follows (undefined groups are the same as those described in any embodiment of the present application),

[0177] In some embodiments, R is wherein Ring A is selected from a 5-6 membered aliphatic ring, aliphatic heterocyclic ring, aromatic ring or heteroaromatic ring, and Ring A is optionally substituted by one or more substituents selected from halogen, haloalkyl, alkyl, alkenyl, alkynyl, alkoxy, phenolic hydroxyl, cyano or formate, preferably selected from halogen, halo(C1-C 12 )alkyl, (C1-C 12 )alkyl, (C2-C 12 )alkenyl, (C3-C5)alkynyl, (C1-C2)alkoxy, phenolic hydroxyl, cyano or (C1-C2)formate.

[0178] In some embodiments, the substituent is one or more halogens, such as F, Cl, Br, I; in some embodiments, the halogen atom is F; in some embodiments, the halogen atom is Cl; in some embodiments, the halogen atom is Br. In some embodiments, the halogen atom is I; in the following embodiments, the substituent is a haloalkyl group, wherein the halogen atom is selected from F, Cl, Br, I; in some embodiments, the substituent is (C1-C 12 )alkyl, (C2-C 12 )alkenyl, (C3-C5)alkynyl, (C1-C2)alkoxy, phenolic hydroxyl, cyano or (C1-C2)formate.

[0179] In certain preferred embodiments of the present invention, certain groups in the compound shown in I are defined as follows (undefined groups are the same as those described in any embodiment of the present application),

[0180] In some embodiments, Ring A is selected from one or more of the following ring systems:

[0181] In some embodiments, when Ring A is When R is Y is Z is a bond, A is H,

[0182] The formula (I) has the following structure

[0183]

[0184] In some embodiments, when Ring A is When R is Y is Z is a bond, A is H,

[0185] The formula (I) has the following structure

[0186]

[0187] In some embodiments, when Ring A is When R is Y is Z is a bond, A is H,

[0188] The formula (I) has the following structure

[0189]

[0190] In certain preferred embodiments of the present invention, certain groups in the compound shown in I are defined as follows (undefined groups are the same as those described in any scheme of this application).

[0191] In some embodiments, R is The B ring is selected from a 5-6 membered aliphatic ring, an aliphatic heterocyclic ring, an aromatic ring or a heteroaromatic ring.

[0192] Ring B is optionally substituted by one or more substituents selected from halogen, haloalkyl, alkyl, alkenyl, alkynyl, alkoxy, phenolic hydroxyl, cyano or formate, preferably selected from halogen, halo (C1-C 12 )alkyl, (C1-C 12 )alkyl, (C2-C 12 )alkenyl, (C3-C5)alkynyl, (C1-C2)alkoxy, phenolic hydroxyl, cyano or (C1-C2)formate.

[0193] In some embodiments, the substituent is one or more halogens, such as F, Cl, Br, I; in some embodiments, the halogen atom is F; in some embodiments, the halogen atom is Cl; in some embodiments, the halogen atom is Br. In some embodiments, the halogen atom is I; in the following embodiments, the substituent is a haloalkyl group, wherein the halogen atom is selected from F, Cl, Br, I; in some embodiments, the substituent is (C1-C 12 )alkyl, (C2-C 12 )alkenyl, (C3-C5)alkynyl, (C1-C2)alkoxy, phenolic hydroxyl, cyano or (C1-C2)formate.

[0194] X is selected from CH2, O, S, (C=O),

[0195] In certain preferred embodiments of the present invention, certain groups in the compound shown in I are defined as follows (undefined groups are the same as those described in any scheme of this application).

[0196] In some embodiments, Ring B is selected from one or more of the following ring systems:

[0197] In some embodiments, when Ring B is When X is O and R is Y is Z is a bond, A is H,

[0198] The formula (I) has the following structure

[0199]

[0200] In some embodiments, when Ring B is When X is R is Y is Z is a bond, A is H,

[0201] The formula (I) has the following structure

[0202]

[0203] In some embodiments, when Ring B is When X is R is Y is Z is a bond, A is H,

[0204] The formula (I) has the following structure

[0205]

[0206] In certain preferred embodiments of the present invention, certain groups in the compound shown in I are defined as follows (undefined groups are the same as those described in any scheme of this application).

[0207] In some embodiments, R is wherein the C ring is selected from H, a 5-6 membered aromatic ring or an aromatic heterocyclic ring, and the C ring is optionally substituted by one or more substituents selected from halogen, haloalkyl, alkyl, alkenyl, alkynyl, alkoxy, phenolic hydroxyl, cyano or formate, preferably selected from halogen, halo (C1-C 12 )alkyl, (C1-C 12 )alkyl, (C2-C 12 )alkenyl, (C3-C5)alkynyl, (C1-C2)alkoxy, phenolic hydroxyl, cyano or (C1-C2)formate.

[0208] In some embodiments, the substituent is one or more halogens, such as F, Cl, Br, I; in some embodiments, the halogen atom is F; in some embodiments, the halogen atom is Cl; in some embodiments, the halogen atom is Br. In some embodiments, the halogen atom is I; in the following embodiments, the substituent is a haloalkyl group, wherein the halogen atom is selected from F, Cl, Br, I; in some embodiments, the substituent is (C1-C 12 )alkyl, (C2-C 12 )alkenyl, (C3-C5)alkynyl, (C1-C2)alkoxy, phenolic hydroxyl, cyano or (C1-C2)formate.

[0209] In some embodiments, the substituent is one or more halogens, such as F, Cl, Br, I; in some embodiments, the halogen atom is F; in some embodiments, the halogen atom is Cl; in some embodiments, the halogen atom is Br. In some embodiments, the halogen atom is I; in the following embodiments, the substituent is a haloalkyl group, wherein the halogen atom is selected from F, Cl, Br, I; in some embodiments, the substituent is (C1-C 12 )alkyl, (C2-C 12 )alkenyl, (C3-C5)alkynyl, (C1-C2)alkoxy, phenolic hydroxyl, cyano or (C1-C2)formate.

[0210] In certain preferred embodiments of the present invention, certain groups in the compound shown in I are defined as follows (undefined groups are the same as those described in any scheme of this application).

[0211] In some embodiments, Ring C is selected from the following ring systems:

[0212] In some embodiments, Ring C is pyridine.

[0213] In some embodiments, when ring C is pyridine, R is Y is

[0214] Z is a bond, A is H,

[0215] The formula (I) has the following structure

[0216]

[0217] In some embodiments, when ring C is pyridine, R is Y is

[0218] Z is a bond, A is H,

[0219] The formula (I) has the following structure

[0220]

[0221] In some embodiments, when ring C is pyridine, R is Y is

[0222] Z is a bond, A is H,

[0223] The formula (I) has the following structure

[0224]

[0225] In some preferred embodiments, the compound of formula (I) of the present invention is preferably the following compound:

[0226] 4-((9H-carbazol-9-yl)methyl)-N'-ethylbenzohydrazide;

[0227] N'-ethyl-4-((1,2,3,4-tetrahydro-9H-carbazol-9-yl)methyl)benzohydrazide;

[0228] 4-((9H-pyrido[2,3-b]indol-9-yl)methyl)-N'-ethylbenzohydrazide;

[0229] 4-((9H-pyrido[3,4-b]indol-9-yl)methyl)-N'-ethylbenzohydrazide;

[0230] 4-((5H-pyrido[4,3-b]indol-5-yl)methyl)-N'-ethylbenzohydrazide;

[0231] 4-(Acridin-10(9H)-ylmethyl)-N'-ethylbenzohydrazide;

[0232] 4-((10H-Benzoxazin-10-yl)methyl)-N'-ethylbenzohydrazide;

[0233] 4-((10H-phenothiazin-10-yl)methyl)-N'-ethylbenzohydrazide;

[0234] 4-((10H-benzo[b]pyrido[2,3-e][1,4]thiazin-10-yl)methyl)-N'-ethylbenzohydrazide;

[0235] 4-((10H-benzo[b]pyrido[2,3-e][1,4]thiazin-10-yl)methyl)-N'-methylbenzohydrazide;

[0236] 4-((10,11-dihydro-5H-dibenzo[b,f]azepin-5-yl)methyl)-N'-ethylbenzohydrazide;

[0237] 4-((5H-dibenzo[b,f]azepin-5-yl)methyl)-N'-ethylbenzohydrazide;

[0238] 4-((11H-benzo[b]pyrido[4,3-f]heptidin-11yl)methyl)-N'-ethylbenzohydrazide;

[0239] N'-ethyl-4-((10-oxo-10,11-dihydro-5H-dibenzo[b,f]heptidin-5-yl)methyl)benzohydrazide;

[0240] 4-(((2,4-Dimethyl-5H-benzo[b]pyrimidin[4,5-f]azepin-5-yl]methyl)methyl)-N'-ethylbenzohydrazide;

[0241] N'-ethyl-4-((3-(trifluoromethyl)-5H-dibenzo[b,f]azepin-5-yl)methyl)benzohydrazide;

[0242] N'-ethyl-4-((10-hydroxy-10,11-dihydro-5H-dibenzo[b,f]azepin-5-yl)methyl)benzohydrazide;

[0243] N'-ethyl-4-((3-methyl-5H-dibenzo[b,f]azepin-5-yl)methyl)benzohydrazide;

[0244] N'-ethyl-4-((3-fluoro-5H-dibenzo[b,f]azepin-5-yl)methyl)benzohydrazide;

[0245] N-(4-(2-ethylhydrazine-1-carbonyl)benzyl)benzamide;

[0246] 1-Butyl-1-(4-(2-ethylhydrazine-1-carbonyl)benzyl)-3-phenylurea;

[0247] N-Butyl-N-(4-(2-ethylhydrazine-1-carbonyl)benzyl)benzamide;

[0248] 1-(4-(2-ethylhydrazine-1-carbonyl)phenyl)-3-(quinolin-8-yl)urea;

[0249] 1-(4-(2-ethylhydrazine-1-carbonyl)benzyl)-3-(quinolin-8-yl)urea;

[0250] N'-ethyl-4-((5-(thiophen-2-yl)-1H-tetrazol-1-yl)methyl)benzohydrazide;

[0251] N'-ethyl-2-(pyridin-3-yl)thiazole-4-carbohydrazide;

[0252] N'-ethyl-4-((quinolin-8-ylamino)methyl)benzohydrazide;

[0253] 4-((2,4-dioxy-3-phenethyl-3,4-dihydroquinazolin-1(2H)-yl)methyl)-N'-ethylbenzohydrazide;

[0254] N'-ethyl-4-((5-(4-methoxybenzoyl)-1H-indol-1-yl)methyl)benzohydrazide;

[0255] 4-((4',5'-dihydrospiro[piperidin-4,7'-thieno[2,3-c]pyran]-1-yl)methyl)-N'-ethylbenzohydrazide;

[0256] 2-(4-bromophenyl)-N'-ethyloxazole-4-carbohydrazide;

[0257] 8-(2-ethylhydrazinyl)-8-oxo-N-phenyloctanamide;

[0258] N-(4-(2-methylhydrazine-1-carbonyl)benzyl)benzamide; or

[0259] 8-(2-Methylhydrazinyl)-8-oxo-N-phenyloctanamide.

[0260] The present invention also provides a method for preparing the compound of formula (I), comprising the following steps:

[0261]

[0262] In some embodiments, during reaction 1, intermediate 1 is hydrolyzed to methyl ester to obtain intermediate 2. The reaction conditions and operations may be conventional conditions and operations in the art. The preferred reaction conditions in the present invention are: KOH, MeOH, reflux, 2h.

[0263] In some embodiments, during reaction 2, intermediate 2 is condensed with tert-butyl 1-ethylhydrazinecarboxylate or tert-butyl 1-methylhydrazinecarboxylate under the action of HATU to obtain a key hydrazide intermediate 3. The reaction conditions and operations may be conventional conditions and operations in the art. The preferred reaction conditions in the present invention are: HATU, DIPEA, DMF, rt, 6h.

[0264] In some embodiments, during reaction 3, intermediate 3 is deprotected from the Boc protecting group to obtain compound (I). The reaction conditions and operations may be conventional conditions and operations in the art. The preferred reaction conditions in the present invention are: TFA, DCM, rt, 2h.

[0265] In some embodiments, intermediate 1 can be obtained by reacting a polycyclic compound containing an aromatic secondary amine with methyl 4-(bromomethyl)benzoate in the presence of NaH to obtain intermediate 1. The reaction conditions are: methyl 4-(bromomethyl)benzoate, NaH, DMF, 0°C - rt, 4h. One synthetic route is shown below:

[0266]

[0267] In some embodiments, intermediate 1 can be obtained by the following method: a compound containing an aromatic amino group is reacted with triphosgene under alkaline conditions to form an isocyanate intermediate. The reaction conditions are: (a) triphosgene, TEA, DCE, 0°C-70°C, 4h, and then reacted with substituted or unsubstituted methyl 4-aminomethylbenzoate hydrochloride or methyl 4-aminobenzoate to obtain intermediate 1. The reaction conditions are: substituted or unsubstituted methyl 4-aminomethylbenzoate hydrochloride, TEA, DCE, 0°C-80°C. One synthetic route is shown below:

[0268]

[0269] In some embodiments, intermediate 1 can be obtained by condensing an aryl formic acid compound with substituted or unsubstituted 4-aminomethylbenzoic acid methyl ester hydrochloride or 4-aminobenzoic acid methyl ester in the presence of HATU to obtain intermediate 1. The reaction conditions are: substituted or unsubstituted 4-aminomethylbenzoic acid methyl ester hydrochloride, HATU, DIPEA, DMF, rt, 6h. One synthetic route is shown below:

[0270]

[0271] In some embodiments, Intermediate 1 is commercially available.

[0272] The present invention also provides a compound of formula (I) as described above or a pharmaceutically acceptable salt thereof for use as a medicament.

[0273] The present invention provides a pharmaceutical composition comprising a compound of formula (I) as described above or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable diluent or carrier.

[0274] Pharmaceutical compositions can be formulated for specific routes of administration, such as oral administration, parenteral administration, and rectal administration. In addition, the pharmaceutical compositions of the present invention can be prepared in solid form (including, but not limited to, capsules, tablets, pills, granules, powders, or suppositories) or in liquid form (including, but not limited to, solutions, suspensions, or emulsions). Pharmaceutical compositions can undergo conventional pharmaceutical operations (e.g., sterilization) and / or can contain conventional inert diluents, lubricants, or buffers, as well as excipients, such as preservatives, stabilizers, wetting agents, emulsifiers, and buffers.

[0275] Typically, the pharmaceutical composition is a tablet or capsule containing the active ingredient and

[0276] a) diluents, such as lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine, etc.;

[0277] b) Lubricants, such as silicon dioxide, talc, stearic acid, its magnesium or calcium salts and / or polyethylene glycol; for tablets also

[0278] c) binders, such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and / or polyvinylpyrrolidone; if desired,

[0279] d) disintegrants, for example starch, agar, alginic acid or its sodium salt, or effervescent mixtures; and / or

[0280] e) absorbents, colorants, flavorings and sweeteners.

[0281] Tablets may be film coated or enteric coated according to methods known in the art.

[0282] Suitable compositions for oral administration include an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof in the form of tablets, lozenges, water or oil suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. Compositions for oral use are prepared according to any method known in the art for preparing pharmaceutical compositions, and in order to provide a refined and palatable formulation, the composition can contain one or more agents selected from sweeteners, flavorings, coloring agents, and preservatives. Tablets can contain active ingredients mixed with non-toxic, pharmaceutically acceptable excipients suitable for preparing tablets. These excipients are, for example, inert diluents (e.g., calcium carbonate, sodium carbonate, lactose, calcium phosphate, or sodium phosphate); granulating agents and disintegrants (e.g., corn starch, or alginic acid); adhesives (e.g., starch, gelatin, or gum arabic); and lubricants (e.g., magnesium stearate, stearic acid, or talc). Tablets are uncoated or coated by known technology to delay disintegration and absorption in the gastrointestinal tract, thereby providing a lasting effect over a long period of time. For example, a time-delay material such as glyceryl monostearate or glyceryl distearate can be used. Preparations for oral administration can be presented in hard gelatin capsules, wherein the active ingredient is mixed with an inert solid diluent (such as calcium carbonate, calcium phosphate or kaolin), or in soft gelatin capsules, wherein the active ingredient is mixed with water or an oil medium (such as peanut oil, liquid paraffin or olive oil).

[0283] Certain injectable compositions are isotonic aqueous solutions or suspensions, and suppositories are advantageously prepared from fat emulsions or suspensions. The compositions may be sterilized and / or contain excipients such as preservatives, stabilizers, wetting or emulsifying agents, solubility promoters, salts for regulating osmotic pressure, and / or buffers. Furthermore, they may contain other therapeutically valuable substances. The compositions are prepared according to conventional mixing, granulation, or coating methods and contain approximately 0.1-75% or approximately 1-50% of the active ingredient.

[0284]

[00145] This invention also provides anhydrous pharmaceutical compositions and dosage forms comprising the compounds of the invention as active ingredients, because water may facilitate the degradation of certain compounds.

[0285] Anhydrous pharmaceutical compositions and dosage forms of the present invention can be prepared using anhydrous or low-water content ingredients and low-water content or low humidity conditions. Anhydrous pharmaceutical compositions can be prepared and stored to maintain their anhydrous properties. Therefore, the anhydrous composition is packaged using known materials that prevent contact with water so that it can be included in a suitable formula kit. Examples of suitable packaging include, but are not limited to, airtight foil, plastics, unit dose containers (e.g., vials), blister packs, and strip packs.

[0286] The present invention further provides pharmaceutical compositions and dosage forms comprising one or more agents that reduce the rate of decomposition of the active ingredient of the compound of the present invention. Such agents (referred to herein as "stabilizers") include, but are not limited to, antioxidants (e.g., ascorbic acid), pH buffers, or salt buffers.

[0287] For an individual of about 50-70 kg, the pharmaceutical composition or combination product of the present invention can be a unit dose of about 1-1000 mg of active ingredient, or about 1-500 mg or about 1-250 mg or about 1-150 mg or about 0.5-100 mg, or about 1-50 mg of active ingredient. The therapeutically effective dose of the compound, pharmaceutical composition, or combination product thereof depends on the species, weight, age, and individual condition of the individual, the condition or disease being treated, or the severity thereof. A physician, clinician, or veterinarian of ordinary skill can readily determine the effective amount of each active ingredient required to prevent, treat, or inhibit the development of a condition or disease.

[0288] The present invention also provides use of the compound of formula (I) or a pharmaceutically acceptable salt thereof in the preparation of an HDAC6 inhibitor.

[0289] The present invention also provides use of the compound of formula (I) or a pharmaceutically acceptable salt thereof in the preparation of a drug for a disease associated with abnormal expression of HDAC6 activity.

[0290] In some embodiments, the disease associated with abnormal expression of HDAC6 activity includes tumors, neurodegenerative diseases, nervous system diseases, stroke, inflammation or autoimmune diseases.

[0291] In some embodiments, the tumor comprises: cancer, tumor growth, colon cancer, breast cancer, bone cancer, brain cancer and other cancers (e.g., osteosarcoma, neuroblastoma, colon adenocarcinoma), chronic myeloid leukemia (CML), acute myeloid leukemia (AML), acute promyelocytic leukemia (APL), cardiac cancer (sarcoma, myxoma, rhabdomyomas, fibromas, lipomas, and teratomas), lung cancer (e.g., bronchogenic carcinoma, alveolar carcinoma, bronchial adenoma, sarcoma, lymphoma, chondromatous hamartoma), shamartoma), mesothelioma), gastrointestinal cancer (e.g., esophageal cancer, gastric cancer, pancreatic cancer, small intestine cancer, large intestine cancer), genitourinary tract cancer (e.g., kidney cancer, bladder cancer and urethra cancer, prostate cancer, testicular cancer), liver cancer (e.g., hepatocellular carcinoma, bile duct cancer, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma), bone cancer (e.g., osteogenic sarcoma, fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma), malignant lymphoma, multiple myeloma, malignant giant cell tumor, chordoma, chondrosteoma, benign chordoma, chondroblastoma, condromixofibroma, osteoid osteoma), The present invention also includes the following types of cancer: osteoma), nervous system tumors (e.g., skull tumor, meningioma, brain tumor, spinal cord tumor), gynecological tumors (e.g., uterine tumor, cervical tumor, ovarian tumor, vulvar and vaginal tumor), hematological cancers (e.g., hematoma, Hodgkin's disease, non-Hodgkin's disease), skin cancers (e.g., malignant melanoma, basal cell carcinoma, malignant squamous cell tumor, Kaposi's sarcoma, dysplastic naevus, lipoma, hemangioma, dermatofibroma, keloid, psoriasis) and adrenal tumors (e.g., neuroblastoma).

[0292] In some embodiments, the neurodegenerative disease comprises Wilson's disease, spinocerebellar disorders, prion diseases, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis (ALS), amyloidosis, Alzheimer's disease, Alexander's disease, alcoholic liver disease, cystic fibrosis, Pick's disease, spinal muscular atrophy, and Lewy body dementia.

[0293] In some embodiments, the autoimmune disease or condition: diabetes, arthritis (including rheumatoid arthritis

[0294] Rheumatoid arthritis, juvenile rheumatoid arthritis, osteoarthritis, psoriatic arthritis, multiple sclerosis, myasthenia gravis, systemic lupus erythematosus, autoimmune thyroiditis, dermatitis (including atopic dermatitis and eczematous dermatitis), psoriasis, Sjögren's syndrome (including keratoconjunctivitis sicca secondary to Sjögren's syndrome), alopecia areata, allergic reactions to arthropod bites, Crohn's disease disease), gastric ulcer, iritis, conjunctivitis, keratoconjunctivitis, ulcerative colitis, asthma, allergic asthma, cutaneous lupus erythematosus, scleroderma, vaginitis, proctitis, drug reactions, leprosy, lupus erythematosus, autoimmune uveitis, allergic encephalomyelitis, acute necrotizing hemorrhagic encephalopathy, progressive bilateral idiopathic hearing loss, aplastic anemia, anemia, idiopathic thrombocytopenia, polychondritis, Wegener's granulomatosis, chronic active hepatitis, Stevens-Johnson syndrome, idiopathic sprues, lichen planus, Graves' ophthalmopathy, sarcoidosis, primary biliary cirrhosis, posterior uveitis, and interstitial pulmonary fibrosis.

[0295] The diseases associated with abnormal expression of HDAC6 activity further include one or more of the following diseases: rheumatoid spondylitis, post-ischemic reperfusion injury, enteritis, chronic inflammatory lung disease, eczema, asthma, acute respiratory distress syndrome, infectious arthritis, chronic progressive arthritis, osteoarthritis, post-traumatic arthritis, gouty arthritis, Reiter syndrome, acute synovitis, acute spondylitis, glomerulonephritis, hemolytic anemia, aplastic anemia, neutropenia, graft-versus-host disease (GVHD), transplant rejection, chronic thyroiditis, Grave's disease, biliary primary cirrhosis, contact dermatitis, sunburn, chronic renal failure, Guillain-Barre syndrome syndrome), uveitis, otitis media, periodontal disease, pulmonary fibrosis, bronchitis, sinusitis, pneumoconiosis, failing lung syndrome, emphysema, pulmonary fibrosis, silicosis, or chronic inflammatory lung disease.

[0296] In some embodiments, a method for treating or preventing a disease associated with abnormal expression of HDAC6 activity is provided, comprising administering a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof to a subject in need thereof.

[0297] In some embodiments, the method comprises administering a compound of Formula (I), or a pharmaceutically acceptable salt thereof, in combination with one, two, three, or four other therapeutic agents.

[0298] As used herein, if the term "combination" is used to describe administration in combination, it will be understood that this can mean simultaneous administration, separate administration, or sequential administration. In one aspect of the invention, "administered in combination" refers to simultaneous administration. In another aspect of the invention, "administered in combination" refers to separate administration. In another aspect of the invention, "administered in combination" refers to sequential administration. When administered sequentially or separately, a delay in administering the second component should not, for example, result in a loss of the benefit of the effect produced by the combination.

[0299] Definition of terms

[0300] The prefix "C x -C y " means that the subsequent group has x (e.g. 1) to y (e.g. 12) carbon atoms, one or more of which may be replaced by one or more heteroatoms or heteroatom groups in certain groups. For example, "(C1-C 12)"alkyl" means that the alkyl group has 1 to 12 carbon atoms. Similarly, the term "xy-membered" ring, where x and y are numerical ranges, such as "5- to 6-membered heterocycle", refers to a ring containing xy atoms (e.g., 5-6), up to 80% of which can be heteroatoms such as N, O, S, and the remaining atoms are carbon.

[0301] "Alkyl" refers to any group derived from a straight or branched saturated hydrocarbon, which can be arbitrarily substituted with one, two or three substituents. Unless otherwise specified, the term "alkyl" is intended to include saturated, unsaturated and partially unsaturated aliphatic groups. When specifically representing an unsaturated group, the term "alkenyl" or "alkynyl" is used. When only a saturated group is represented, the term "alkyl" is used. Preferably, alkyl includes but is not limited to methyl, ethyl, propyl such as propyl-1-yl, propyl-2-yl (isopropyl), butyl such as butyl-1-yl (n-butyl), butyl-2-yl (sec-butyl), 2-methyl-propyl-1-yl (isobutyl), 2-methyl-propyl-2-yl (tert-butyl), pentyl, hexyl, octyl, decyl, etc. Unless otherwise specified, alkyl has 1 to 12 carbon atoms, for example, 1 to 8 carbon atoms, for example, 1 to 6 carbon atoms, for example, 1 to 4 carbon atoms, for example, 1 to 2 carbon atoms.

[0302] "Alkenyl" refers to an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon double bond, "(C2-C 12 The term "(C2-C6)alkenyl" refers to a straight or branched alkenyl group containing 2 to 12 carbon atoms, including but not limited to vinyl, 1-propenyl, 2-propenyl, 1-, 2- or 3-butenyl, etc., preferably "(C2-C6)alkenyl", more preferably "(C2-C4)alkenyl". The alkenyl group may be substituted or unsubstituted.

[0303] "Alkynyl" refers to an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon triple bond, "(C2-C 12 The term "(C2-C8)alkynyl" refers to a straight or branched chain alkynyl group containing 2 to 12 carbon atoms, including but not limited to ethynyl, 1-propynyl, 2-propynyl, 1-, 2- or 3-butynyl, preferably "(C2-C8)alkynyl", more preferably "(C3-C6)alkynyl". The alkynyl group may be substituted or unsubstituted.

[0304] "Ylene" refers to a divalent group, such as alkylene refers to a divalent alkyl group, alkenylene refers to a divalent alkenyl group, alkynylene refers to a divalent alkynyl group, cycloalkylene refers to a divalent cycloalkyl group, heterocycloalkylene refers to a divalent heterocycloalkyl group, arylene refers to a divalent aryl group, and heteroarylene refers to a divalent heteroaryl group. The alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups are as defined above, and the alkylene group may be optionally substituted or unsubstituted.

[0305] "Alkylene" is an alkyl group positioned between and used to connect two other chemical groups, and alkyl is defined as above. The term "alkylene" is intended to include saturated, unsaturated, and partially unsaturated aliphatic groups. When unsaturated groups are specifically intended, the terms "alkenylene" or "alkynylene" are used. When only saturated groups are intended, the term "alkylene" is used. The term "alkylene" includes descriptors of ranges of carbon atoms, such as (C1-C 12 ) alkylene, the number of carbon atoms refers to the length of the linear chain connecting the two chemical groups between which the alkylene is located. As shown below, any carbon atom on the alkylene may be optionally substituted, and the substituent may contain additional carbon atoms, such as, but not limited to,

[0306] —CH2CH2 CH2CH2—, —CH2CH=CHCH2—, —CH2≡CCH2—,

[0307] —CH2CH2CH(CH2CH2CH3)CH2—.

[0308] "Haloalkyl" refers to an alkyl group in which one or more hydrogen atoms are each replaced by a halogen. Halogens include fluorine, chlorine, bromine, or iodine. Examples include, but are not limited to, -CH2Cl, CH2F, -CH2Br, CHF2, CF3, -CFClBr, CH2CH2F, -CH2CH2Cl, CH2CHF2, CH2CF3, -CH2CCl3, CH2CH2CH2F, CH2CH2CHF2, CH2CH2CF3, and the like, as well as alkyl groups such as perfluoroalkyl groups in which all hydrogen atoms are replaced by fluorine atoms.

[0309] "Alkoxy" refers to a portion of the formula -O-alkyl, wherein the alkyl portion is as defined above. Non-limiting examples include methoxy, ethoxy, propoxy, butoxy, and the like. The alkoxy group may be optionally substituted or unsubstituted. For example, (C1-C2)alkoxy refers to an alkyl portion having 1-2 carbon atoms attached to an oxygen.

[0310] "Heterocycle" refers to a single saturated or partially unsaturated non-aromatic ring or non-aromatic polycyclic ring system having at least one heteroatom (i.e., at least one ring heteroatom selected from oxygen, nitrogen, and sulfur) in the ring. Unless otherwise indicated, a heterocyclic radical has 5 to 14 ring atoms, such as 3 to 10 ring atoms, such as 5 to 10 ring atoms, or such as 5 to 6 ring atoms. Thus, the term includes a single saturated or partially unsaturated ring (e.g., 3, 4, 5, 6, or 7-membered ring) having about 1 to 6 ring carbon atoms and about 1 to 3 ring heteroatoms selected from oxygen, nitrogen, and sulfur in the ring. When valence requirements permit, the rings of multiple fused rings (e.g., bicyclic heterocyclic radicals) systems can be interconnected by fusion, spirocycles, and bridge bonds. Heterocycles include, but are not limited to, groups derived from oxiranyl, thiolyl, aziridine, imidazolidine, oxetanyl, thietanyl, azetidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperidinyl, pyrazolidine, pyrrolidone dihydropyridine, tetrahydropyridine, 1,4-dioxanyl, 1,4-oxathianyl, morpholinyl, 1,4-dithianyl , piperazinyl, 1,4-azathiolanyl, azepanyl, thiepanyl, azepanyl, 1,4-dioxepanyl, 1,4-oxathiepanyl, 1,4-azathiepanyl, 1,4-dithiepanyl, 1,4-azathiepanyl, 1,4-diazepanyl, tropanyl, tetrahydro-2H-thiopyran 1,1-dioxide, quinuclidinyl, N-bromopyrrolidine, N-chloropiperidine. Heterocyclic rings include spirocycles such as aza or oxo-spiroheptane. Heterocyclic groups also include partially unsaturated ring systems having one or more double bonds, including fused ring systems having one aromatic ring and one non-aromatic ring, but not a completely aromatic ring system. Examples include dihydroquinolines such as 3,4-dihydroquinoline, dihydroisoquinolines such as 1,2-dihydroisoquinoline, dihydroimidazoles, tetrahydroimidazoles, etc., indolines, isoindolines, isoindolinones (e.g., isoindolin-1-one), isatin, dihydrophthalazines, quinolinones, spiro[cyclopropane-1,1′-isoindoline]-3′-ones, etc. Other examples of heterocycles include, for example, 3,8-diazabicyclo[3.2.1]octyl, 2,5-diazabicyclo[2.2.1]heptyl, 3,6-diazabicyclo[3.1.1]heptyl, 3-oxa-7,9-diazabicyclo[3.3.1]nonyl, and hexahydropyrazino[2,1-c][1,4]oxazinyl.

[0311] "Multiple" independently refers to 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0312] "Optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs or does not occur. For example, "optionally substituted with one or more substituents" means that the substituents may but need not be present, and that the description includes instances where the heterocyclic group is substituted with substituents and instances where the heterocyclic group is not substituted with substituents.

[0313] "Substituted" means that one or more hydrogen atoms, preferably up to five, and more preferably one to three, hydrogen atoms in a group are independently replaced by a corresponding number of substituents. It goes without saying that substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) which substitutions are possible or impossible without undue effort. For example, an amino or hydroxyl group with free hydrogen may be unstable when combined with a carbon atom with an unsaturated (e.g., olefinic) bond.

[0314] It will be appreciated by those skilled in the art that salts of compounds of formula (I) can be prepared, including pharmaceutically acceptable salts. These salts can be prepared in situ during the final separation and purification of the compound, or by independently reacting the purified compound in its free acid or free base form with a suitable base or acid.

[0315] Pharmaceutically acceptable acid addition salts can be formed with inorganic and organic acids, for example, acetate, aspartate, benzoate, benzenesulfonate, bromide / hydrobromide, bicarbonate / carbonate, bisulfate / sulfate, camphorsulfonate, chloride / hydrochloride, citrate, edisylate, fumarate, glucoheptonate, gluconate, glucuronate, hippurate, hydroiodide / iodide, isethionate, lactate, lactobionate, lauryl sulfate, malate, maleate, malonate, mandelate, methanesulfonate, methylsulfate, naphthoate, naphthylate, nicotinate, nitrate, stearate, oleate, oxalate, palmitate, pamoate, phosphate / hydrogenphosphate / dihydrogenphosphate, polygalacturonate, propionate, stearate, succinate, sulfosalicylate, tartrate, toluenesulfonate, and trifluoroacetate.

[0316] Inorganic acids which can form salts include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like.

[0317] Organic acids which can form salts include, for example, acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, sulfosalicylic acid, etc. Pharmaceutically acceptable base addition salts can be formed with inorganic or organic bases.

[0318] Inorganic bases from which salts can be formed include, for example, ammonium salts and metals from Groups I to XII of the periodic table. In certain embodiments, the salts are derived from sodium, potassium, ammonium, calcium, magnesium, iron, silver, zinc, and copper; particularly suitable salts include ammonium, potassium, sodium, calcium, and magnesium salts.

[0319] Organic bases from which salts can be formed include, for example, primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, basic ion exchange resins, and the like. Certain organic amines include isopropylamine, diethanolamine, diethylamine, lysine, meglumine, piperazine, and tromethamine.

[0320] The pharmaceutically acceptable salts of the present invention can be synthesized from alkaline or acidic moieties by conventional chemical methods. Typically, these salts can be prepared by reacting the free acid forms of these compounds with a chemical amount of a suitable base (a hydroxide, carbonate, bicarbonate, etc. of Na, Ca, Mg or K), or by reacting the free base forms of these compounds with a chemical amount of a suitable acid. These reactions are typically carried out in water or in an organic solvent, or in a mixture of the two. Typically, when appropriate, a non-aqueous medium such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile is used. Lists of other suitable salts can be found in "Remington's Pharmaceutical Sciences", 20th edition, Mack Publishing Company, Easton, Pa., (1985); and in "Handbook of Pharmaceutical Salts: Properties, Selection, and Use" by Stahl and Wermuth (Wiley-VCH, Weinheim, Germany, 2002).

[0321] Solvates of compounds of formula (I) can also be prepared, including pharmaceutically acceptable solvates. "Solvate" refers to a complex of variable chemical amounts formed by a solute and a solvent. Such solvents for the purposes of the present invention do not affect the biological activity of the solute. Examples of suitable solvents include, but are not limited to, water, MeOH, EtOH, and AcOH. Solvates in which water is a solvent molecule generally refer to hydrates. Hydrates include components comprising a stoichiometric amount of water, as well as components comprising a variable amount of water.

[0322] As used herein, the term "pharmaceutically acceptable" means a compound that is suitable for pharmaceutical use. Salts and solvates (e.g., hydrates and hydrates of salts) of the compounds of the invention that are suitable for pharmaceutical use are those in which the counterion or associated solvent is pharmaceutically acceptable. However, salts and solvates having non-pharmaceutically acceptable counterions or associated solvents are also within the scope of the invention, for example, for use as intermediates in the preparation of other compounds of the invention and pharmaceutically acceptable salts and solvates thereof.

[0323] The compound of formula (I) (including its salt and solvate) can exist in crystalline form, non-crystalline form or its mixture. The compound or its salt or solvate can also show polymorphism, i.e. the ability to appear in different crystalline forms. These different crystalline forms are generally known as "polymorphs". Polymorphs have the same chemical composition, but the stacking, geometric arrangement and other descriptive characteristics of the crystalline solid state are different. Therefore, polymorphs can have different physical properties, such as shape, density, hardness, deformability, stability and solubility properties. Polymorphs usually show different melting points, IR spectra and X-ray powder diffraction patterns, all of which can be used for identification. It will be appreciated by those skilled in the art that, for example, different polymorphs may be produced by changing or adjusting the conditions used in the crystallization / recrystallization of the compound of formula (I).

[0324] The present invention also includes different isomers of the compound of formula (I). "Isomer" refers to compounds with the same composition and molecular weight, but different physical and / or chemical properties. The difference in structure can be in the structure (geometric isomers) or in the ability to rotate plane polarized light (stereoisomers). With regard to stereoisomers, the compound of formula (I) can have one or more asymmetric carbon atoms and can appear as a racemate, a racemic mixture, and as a single enantiomer or diastereomer. All such isomeric forms are included within the scope of the present invention, including mixtures thereof. If the compound contains a double bond, the substituent can be an E or Z configuration. If the compound contains a disubstituted cycloalkyl, the substituent of the cycloalkyl can have a cis- or trans-configuration. It is also desirable to include all tautomeric forms.

[0325] Any asymmetric atom (e.g., carbon, etc.) of the compound of formula (I) can exist in racemic or enantiomeric enrichment, such as (R)-, (S)-, or (R,S)-configuration. In certain embodiments, each asymmetric atom is present in at least 50% enantiomeric excess, at least 60% enantiomeric excess, at least 70% enantiomeric excess, at least 80% enantiomeric excess, at least 90% enantiomeric excess, at least 95% enantiomeric excess, or at least 99% enantiomeric excess in the (R)- or (S)-configuration. Where possible, substituents on atoms with unsaturated double bonds are present in cis-(Z)- or trans-(E)-form.

[0326] Thus, as used herein, the compounds of formula (I) can be in the form of one of the possible isomers, rotamers, atropisomers, tautomers or mixtures thereof, for example as substantially pure geometric isomers (cis or trans), diastereomers, optical isomers (enantiomers), racemates or mixtures thereof.

[0327] Any resulting mixtures of isomers can be separated on the basis of the physicochemical differences of the constituents into the pure or substantially pure geometric or optical isomers, diastereomers, racemates, for example, by chromatography and / or fractional crystallization.

[0328] Any resulting racemates of the final products or intermediates can be resolved into the optical enantiomers by known methods (e.g., by separation of their diastereomeric salts), which are obtained with optically active acids or bases and release the optically active acidic or basic compounds. In particular, basic moieties can be used to resolve the compounds of the invention into their optical enantiomers, for example, by fractional crystallization of salts formed with optically active acids (e.g., tartaric acid, dibenzoyltartaric acid, diacetyltartaric acid, di-O,O'-p-toluoyltartaric acid, mandelic acid, malic acid, or camphor-10-sulfonic acid). Racemic products can also be resolved by chiral chromatography, such as high pressure liquid chromatography (HPLC) using a chiral adsorbent.

[0329] The present invention includes the unlabeled form and isotope-labeled form of the compound of formula (I). Isotope-labeled compounds have the structure described by the chemical formula given herein, except that one or more atoms are replaced by atoms with a selected atomic weight or mass number. Examples of isotopes that can be incorporated into the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, such as 2H, 3H, 11C, 13C, 14C, 15N, 18F, 31P, 32P, 35S, 36Cl, 125I, respectively. The present invention includes various isotope-labeled compounds as defined herein, such as those in which radioactive isotopes (such as 3H and 14C) occur or those in which non-radioactive isotopes (such as 2H and 13C) occur. These isotope-labeled compounds can be used for metabolic studies (e.g., using 14C), reaction kinetic studies (e.g., using 2H or 3H), detection or imaging techniques, such as positron emission tomography (PET) or single photon emission computed tomography (SPECT), including drug substrate tissue distribution analysis, or for radiotherapy of patients. In particular, 18F or labeled compounds may be particularly desirable for PET or SPECT studies. Isotope-labeled compounds of formula (I) can generally be prepared by conventional techniques known to those skilled in the art or by methods analogous to those described in the accompanying Examples and Preparations, using appropriate isotope-labeled reagents instead of the unlabeled reagents previously used.

[0330] In addition, substitution with heavier isotopes, particularly deuterium (i.e. 2H or D) may bring certain therapeutic advantages caused by stronger metabolic stability, such as increased in vivo half-life or reduced dosage requirements or improvement of therapeutic index. It will be understood that deuterium is considered as a substituent of a compound of formula (I) in this article. The concentration of this heavier isotope, particularly deuterium, may be determined by the isotopic enrichment factor. As used herein, the term "isotopic enrichment factor" refers to the ratio between the isotopic abundance and the natural abundance of a particular isotope. If a substituent in a compound of the invention is labeled as deuterium, then for each deuterium atom labeled, the compound has an isotopic enrichment factor of at least 3500 (52.5% deuterium incorporation at each deuterium atom labeled), at least 4000 (60% deuterium incorporation), at least 4500 (67.5% deuterium incorporation), at least 5000 (75% deuterium incorporation), at least 5500 (82.5% deuterium incorporation), at least 6000 (90% deuterium incorporation), at least 6333.3 (95% deuterium incorporation), at least 6466.7 (97% deuterium incorporation), at least 6600 (99% deuterium incorporation), or at least 6633.3 (99.5% deuterium incorporation).

[0331] Those skilled in the art will be able to identify whether a stereocenter is present in a compound of formula (I). Therefore, the present invention includes possible stereoisomers and includes both racemic compounds and single enantiomers. When the desired compound is a single enantiomer, it can be obtained by stereospecific synthesis or by splitting the final product or any convenient intermediate. The splitting of the final product, intermediate or starting material can be achieved by any suitable method known in the art. See, for example, "Stereochemistry of Organic Compounds" by E.L. Eliel, S.H. Wilen and L.N. Mander (Wiley-interscience, 1994).

[0332] The present invention is further illustrated below by means of specific preparation examples.

[0333] The names and structures of the embodiments are shown in the following table

[0334]

[0335]

[0336]

[0337] Example

[0338] Example 1

[0339] 4-((9H-carbazol-9-yl)methyl)-N'-ethylbenzohydrazide

[0340]

[0341] Synthesis route as Synthesis route I

[0342] The specific synthesis steps are as follows:

[0343] a. Synthesis of Compound 1 [Methyl 4-((9H-carbazol-9-yl)methyl)benzoate]

[0344] At room temperature, carbazole (500 mg, 3 mmol) was placed in a 100 mL eggplant-shaped flask and dissolved in anhydrous DMF. NaH (144 mg, 6 mmol) was added under ice-cooling and allowed to react for 1 hour. Methyl 4-bromomethylbenzoate (1.03 g, 4.5 mmol) was added under ice-cooling and allowed to react at room temperature for 4 hours. TLC confirmed the reaction was complete, and 30 mL of water was added. The mixture was extracted with ethyl acetate (30 mL x 3). The organic phases were combined and dried over anhydrous NaSO. Column chromatography afforded compound 1 (650 mg, 69% yield) as a white solid, which was used directly in the next step.

[0345] 1 H NMR(400MHz, DMSO-d6)δ8.18-8.11(m,2H),7.83(d,J=8.2Hz,2H),7.60-7.51(m,2H), 7.44-7.35(m,2H),7.27-7.14(m,4H),5.73(s,2H),3.76(s,3H).LC-MS(Waterse2695 HPLC and Waters ACQUITY QDa Mass Detector)m / z:315.94[M+H] + .

[0346] b. Synthesis of Compound 2 [4-((9H-carbazol-9-yl)methyl)benzoic acid]

[0347] At room temperature, methyl 4-((9H-carbazol-9-yl)methyl)benzoate (500 mg, 1.58 mmol) was dissolved in 20 mL of methanol and 10 mL of 3N KOH was added. The mixture was refluxed for 2 hours. TLC confirmed the complete reaction. The organic solvent was evaporated and the pH was adjusted to acidic with 1 M HCl. A solid precipitated, which was filtered and dried to afford compound 2 (450 mg, crude) as a white solid. This was used directly in the next step without further purification.

[0348] LC-MS m / z:300.82[MH] - .

[0349] c. Synthesis of Compound 3 [tert-butyl 2-(4-(((9H-carbazol-9-yl)methyl)benzoyl)-1-ethylhydrazine-1-carboxylate]

[0350] At room temperature, 4-((9H-carbazol-9-yl)methyl)benzoic acid (300 mg, 1 mmol) was placed in a 100 mL eggplant-shaped flask and dissolved in 30 mL of dichloromethane. 1-Ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC·HCl) (230 mg, 1.2 mmol), 1-hydroxybenzotriazole (HOBt) (162 mg, 1.2 mmol), and triethylamine (121 mg, 1.2 mmol) were added under ice-cooling and allowed to react for 0.5 hour. 1-Ethylhydrazine-1-carboxylic acid tert-butyl ester (192 mg, 1.2 mmol) was added and allowed to react overnight at room temperature. TLC confirmed the complete reaction of the starting material. The reaction mixture was then washed with water (20 mL x 3) and the organic phase was dried over anhydrous Na2SO4. Column chromatography afforded compound 3 (90 mg, 65% yield) as a white solid, which was used directly in the next step.

[0351] 1H NMR (400MHz, DMSO-d6) δ10.39(s,1H),8.16(d,J=7.8Hz,2H),7.69(d,J=11.7Hz,2H),7.66-7.54(m,3H),7. 46-7.35(m,2H),7.25-7.10(m,4H),5.71(s,2H),3.44-3.34(m,2H),1.38(s,9H),1.06-0.94(m,3H).LC-MS m / z:443.96[M+H] + .

[0352] d. Example 1 [4-((9H-carbazol-9-yl)methyl)-N'-ethylbenzohydrazide] Synthesis

[0353] At room temperature, tert-butyl 2-(4-(((9H-carbazol-9-yl)methyl)benzoyl)-1-ethylhydrazine-1-carboxylate (250 mg, 0.56 mmol) was placed in a 50 mL eggplant-shaped flask and dissolved in 15 mL of dichloromethane. Then, 5 mL of trifluoroacetic acid was added and allowed to react for 2 hours. TLC confirmed the complete reaction. After evaporating the solvent, the pH was adjusted to alkaline with 1 N NaOH, followed by extraction with dichloromethane (20 mL x 3). The organic phases were combined and dried over anhydrous NaSO. Column chromatography afforded the compound of Example 1 (150 mg, 78% yield) as a white solid.

[0354] 1 H NMR(400MHz,DMSO-d6)δ9.90(s,1H),8.19-8.11(m,2H),7.71-7.65(m,2H),7.58(d,J=8.2Hz,2H),7.44- 7.35(m,2H),7.23-7.13(m,4H),5.68(s,2H),5.02(s,1H),2.73(q,J=7.1Hz,2H),0.96(t,J=7.1Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ165.58,141.63,140.64,132.88,127.91,127.14,126.43,122.80,120.93,119.67,110.03,45.97,45.85,13.59.LC-MS m / z:343.91[M+H] + .

[0355] Example 2

[0356] N'-ethyl-4-((1,2,3,4-tetrahydro-9H-carbazol-9-yl)methyl)benzohydrazide

[0357]

[0358] Synthesis route as Synthesis route I

[0359] The specific synthesis steps are as follows:

[0360] The carbazole in Example 1 was replaced by 2,3,4,9-tetrahydro-1H-carbazole, and the rest was the same as in Example 1. The total yield was 37%.

[0361] 1 H NMR(400MHz, DMSO-d6)δ9.92(s,1H),7.74-7.66(m,2H),7.36(dd,J=7.4,1.5Hz,1H),7.31-7.27(m,1H),7.07-6.90( m,4H),5.33(s,2H),5.03(s,1H),2.75(q,J=7.2Hz,2H),2.68-2.53(m,4H),1.88-1.64(m,4H),0.97(t,J=7.1Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ165.59,142.50,136.68,135.99,132.65,127.91,127.52,126.77, 121.02,119.11,118.02,109.79,109.44,45.99,45.75,23.26,22.11,21.22,13.60.LC-MS m / z:347.91[M+H] + .

[0362] Example 3

[0363] 4-((9H-pyrido[2,3-b]indol-9-yl)methyl)-N'-ethylbenzohydrazide

[0364]

[0365] Synthesis route as Synthesis route I

[0366] The specific synthesis steps are as follows:

[0367] The carbazole in Example 1 was replaced by 9H-pyrido[2,3-b]indole, and the rest was the same as in Example 1. The total yield was 32%.

[0368] 1H NMR(400MHz, DMSO-d6)δ9.89(s,1H),8.55(dd,J=7.7,1.6Hz,1H),8.47(dd,J=4.9,1.6Hz,1H),8.20(d,J=7.7Hz,1H),7.71-7.65(m,2H ),7.57(d,J=8.2Hz,1H),7.49-7.41(m,1H),7.30-7.21(m,4H),5.73(s,2H),5.02(s,1H),2.73(q,J=7.2Hz,2H),0.95(t,J=7.2Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ165.58,151.47,146.74,141.50,139.47,132.89,129.39,127.89,1 27.46,127.42,121.98,120.63,120.50,116.19,115.69,110.60,45.96,44.37,13.59.LC-MS m / z:345.86[M+H] + .

[0369] Example 4

[0370] 4-((9H-pyrido[3,4-b]indol-9-yl)methyl)-N'-ethylbenzohydrazide

[0371]

[0372] Synthesis route as Synthesis route I

[0373] The specific synthesis steps are as follows:

[0374] The carbazole in Example 1 was replaced by 9H-pyrido[3,4-b]indole, and the rest was the same as in Example 1. The total yield was 35%.

[0375] 1 H NMR(400MHz, DMSO-d6)δ9.92(s,1H),9.37(d,J=1.0Hz,1H),8.46(d,J=5.8Hz,1H),8.32-8.23(m,1H),7.74-7.60(m,4H) ,7.51-7.44(m,1H),7.33-7.26(m,1H),7.20(d,J=8.1Hz,2H),5.72(s,2H),2.73(q,J=7.2Hz,2H),0.95(t,J=7.2Hz,3H). 13C NMR(101MHz,DMSO-d6)δ165.51,145.30,144.81,143.15,140.84,140.57,133.06,128.01,1 27.53,127.27,121.49,121.35,121.25,119.62,110.78,105.64,45.99,45.95,13.58.LC-MS m / z:345.89[M+H] + .

[0376] Example 5

[0377] 4-((5H-pyrido[4,3-b]indol-5-yl)methyl)-N'-ethylbenzohydrazide

[0378]

[0379] Synthesis route as Synthesis route I

[0380] The specific synthesis steps are as follows:

[0381] The carbazole in Example 1 was replaced by 5H-pyrido[4,3-b]indole, and the rest was the same as in Example 1. The total yield was 33%.

[0382] 1 H NMR (400MHz, DMSO-d6) δ9.92 (s, 1H), 9.04 (d, J = 1.1Hz, 1H), 8.40-8.36 (m, 1H), 8.33-8.23 (m, 1H), 8.19-8.11 (m, 1H) ,7.75-7.64(m,3H),7.63-7.52(m,1H),7.34-7.18(m,3H),5.80(s,2H),2.72(q,J=7.1Hz,2H),0.95(t,J=7.1Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ165.53,141.40,141.17,139.30,136.67,133.33,133.06,129.14,128.1 5,128.00,127.33,127.28,122.68,121.09,120.44,115.31,110.97,46.15,45.95,13.58.LC-MS m / z:345.88[M+H] + .

[0383] Example 6

[0384] 4-(Acridin-10(9H)-ylmethyl)-N'-ethylbenzohydrazide

[0385]

[0386] Synthesis route as Synthesis route I

[0387] The specific synthesis steps are as follows:

[0388] The carbazole in Example 1 was replaced by acridine, and the rest was the same as in Example 1. The total yield was 41%.

[0389] 1 H NMR (400MHz, DMSO-d6) δ9.86 (s, 1H), 9.07 (s, 1H), 8.21-8.10 (m, 2H), 8.02 (d, J = 8.4Hz, 1H), 7.87-7.79 (m, 1H) ,7.72-7.57(m,4H),7.55-7.44(m,3H),4.98(s,1H),4.71(s,2H),2.72(q,J=7.1Hz,2H),0.95(t,J=7.2Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ165.81,148.26,147.37,145.48,139.51,137.08,131.36,131.08,130.46,1 29.87,129.56,128.93,127.79,127.53,126.86,126.61,126.51,126.28,46.00,37.01,13.59.LC-MS m / z:357.95[M+H] + .

[0390] Example 7

[0391] 4-((10H-Benzoxazin-10-yl)methyl)-N'-ethylbenzohydrazide

[0392]

[0393] Synthesis route as Synthesis route I

[0394] The specific synthesis steps are as follows:

[0395] The carbazole in Example 1 was replaced by 10H-benzoxazine, and the rest was the same as in Example 1. The total yield was 32%.

[0396] 1H NMR (400MHz, DMSO-d6) δ9.95 (s, 1H), 7.79-7.72 (m, 2H), 7.31 (d, J = 8.2Hz, 2H), 6.73-6.60 (m, 6H), 6 .42(dd,J=7.8,1.5Hz,2H),5.06(s,1H),4.89(s,2H),2.76(q,J=7.1Hz,2H),0.98(t,J=7.2Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ165.65,144.63,140.54,133.64,132.62,128.05,126.70,124.59,121.81,115.67,112.93,47.37,46.00,13.61.LC-MS m / z:359.93[M+H] + .

[0397] Example 8

[0398] 4-((10H-phenothiazin-10-yl)methyl)-N'-ethylbenzohydrazide

[0399]

[0400] Synthesis route as Synthesis route I

[0401] The specific synthesis steps are as follows:

[0402] The carbazole in Example 1 was replaced by phenothiazine, and the rest was the same as in Example 1. The yield was 42%.

[0403] 1 H NMR (400MHz, DMSO-d6) δ9.94(s,1H),7.74(d,J=8.2Hz,2H),7.35(d,J=8.1Hz,2H),7.12(dd,J=7.5,1.5Hz,2H),7. 06-6.99(m,2H),6.92-6.83(m,2H),6.78-6.69(m,2H),5.14(s,2H),2.75(q,J=7.2Hz,2H),0.98(t,J=7.1Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ165.59,144.68,141.01,132.50,128.08,127.89,127.41,127.22,123.27,123.19,116.39,51.29,46.01,13.62.LC-MS m / z:375.91[M+H] + .

[0404] Example 9

[0405] 4-((10H-Benzo[b]pyrido[2,3-e][1,4]thiazin-10-yl)methyl)-N'-ethylbenzohydrazide

[0406]

[0407] Synthesis route as Synthesis route I

[0408] The specific synthesis steps are as follows:

[0409] The carbazole in Example 1 was replaced by 10H-benzo[b]pyrido[2,3-e][1,4]thiazine, and the rest was the same as in Example 1. The yield was 46%.

[0410] 1 H NMR (400MHz, DMSO-d6) δ9.92(s,1H),7.90(dd,J=4.9,1.7Hz,1H),7.71(d,J=8.0Hz,2H),7.45(dd,J=7.5,1.7Hz,1H),7.29(d,J=8.0Hz,2H),7.07(dd,J =7.5,1.6Hz,1H),7.01-6.94(m,1H),6.89-6.80(m,2H),6.65(d,J=8.2Hz,1 H), 5.32 (s, 2H), 5.09 (s, 1H), 2.75 (q, J = 7.2Hz, 2H), 0.98 (t, J = 7.2Hz, 3H). 13 C NMR(101MHz,DMSO-d6)δ165.71,154.11,145.77,142.26,141.64,135.17,132.19,128.29,1 27.73,127.26,126.92,123.80,120.31,119.16,116.65,116.16,48.31,46.01,13.62.LC-MS m / z:377.01[M+H] + .

[0411] Example 10

[0412] 4-((10H-Benzo[b]pyrido[2,3-e][1,4]thiazin-10-yl)methyl)-N'-methylbenzohydrazide

[0413]

[0414] Synthesis route as Synthesis route I

[0415] The specific synthesis steps are as follows:

[0416] In Example 9, tert-butyl 1-ethylhydrazine-1-carboxylate was replaced by tert-butyl 1-methylhydrazine-1-carboxylate, and the rest was the same as in Example 9. The total yield was 44%.

[0417] 1 H NMR (400MHz, DMSO-d6) δ9.94(s,1H),7.94-7.85(m,1H),7.70(d,J=8.1Hz,2H),7.46(dd,J=7.5,1.6Hz,1H),7.29(d,J=8.1Hz,2H),7. 07(dd,J=7.6,1.5Hz,1H),7.02-6.94(m,1H),6.91-6.81(m,2H),6.64(dd,J=8.3,1.2Hz,1H),5.32(s,2H),5.06(s,1H),2.47(s,3H). 13 C NMR(101MHz,DMSO-d6)δ165.51,154.11,145.78,142.26,141.65,135.19,132.20,128.3 0,127.68,127.27,126.92,123.81,120.30,119.17,116.66,116.16,48.29,39.09.LC-MS m / z:362.95[M+H] + .

[0418] Example 11

[0419] 4-((10,11-Dihydro-5H-dibenzo[b,f]azepin-5-yl)methyl)-N'-ethylbenzohydrazide

[0420]

[0421] Synthesis route as Synthesis route I

[0422] The specific synthesis steps are as follows:

[0423] The carbazole in Example 1 was replaced by 10,11-dihydro-5H-dibenzo[b,f]azepine, and the rest was the same as in Example 1. The total yield was 46%.

[0424] 1H NMR (400MHz, DMSO-d6) δ9.89(s,1H),7.65(d,J=8.0Hz,2H),7.45(d,J=7.9Hz,2H),7.13(d,J=8.0Hz,2H),7.0 9-6.97(m,4H),6.83(t,J=7.3Hz,2H),5.05(s,1H),4.93(s,2H),2.73(q,J=7.2Hz,2H),0.96(t,J=7.2Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ165.64,148.55,142.55,134.02,132.27,130.22,1 28.53,127.53,126.81,123.08,120.52,54.47,46.00,32.20,13.60.LC-MS m / z:372.03[M+H] + .

[0425] Example 12

[0426] 4-((5H-dibenzo[b,f]azepin-5-yl)methyl)-N'-ethylbenzohydrazide

[0427]

[0428] Synthesis route as Synthesis route I

[0429] The specific synthesis steps are as follows:

[0430] The carbazole in Example 1 was replaced by 5H-dibenzo[b,f]azepine, and the rest was the same as in Example 1. The total yield was 45%.

[0431] 1 H NMR(400MHz,DMSO-d6)δ9.87(s,1H),7.63(d,J=8.3Hz,2H),7.45(d,J=8.0Hz,2H),7.24-7.03(m,6H),6 .99-6.87(m,2H),6.81(s,2H),5.11(s,1H),4.96(s,2H),2.73(q,J=7.1Hz,2H),0.96(t,J=7.1Hz,3H). 13C NMR (101MHz, DMSO-d6) δ165.63,150.89,142.19,133.81,132.69,132.28,129. 46,129.41,128.23,127.49,123.93,121.07,54.06,46.00,38.77,13.58.LC-MS m / z:369.96[M+H] + .

[0432] Example 13

[0433] 4-((11H-Benzo[b]pyrido[4,3-f]heptidin-11yl)methyl)-N'-ethylbenzohydrazide

[0434]

[0435] Synthesis route as Synthesis route I

[0436] The specific synthesis steps are as follows:

[0437] The carbazole in Example 1 was replaced by 11H-benzo[b]pyrido[4,3-f]azepine, and the rest was the same as in Example 1. The total yield was 45%.

[0438] 1 H NMR (400MHz, DMSO-d6) δ9.89(s,1H),8.31(s,1H),8.07(d,J=4.9Hz,1H),7.65(d,J=8.1Hz,2H),7.47(d,J=8.1Hz,2H),7.29-7.21 (m,1H),7.19-7.08(m,2H),7.08-6.92(m,3H),6.77(d,J=11.4Hz,1H),5.05(s,2H),2.73(q,J=7.1Hz,2H),0.96(t,J=7.1Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ165.59,151.20,146.23,144.85,142.82,141.57,140.90,137.20,132.9 6,132.51,130.33,130.29,128.31,127.60,124.30,123.05,121.45,53.79,45.99,13.58.LC-MS m / z:370.94[M+H] + .

[0439] Example 14

[0440] N'-ethyl-4-((10-oxo-10,11-dihydro-5H-dibenzo[b,f]heptidin-5-yl)methyl)benzohydrazide

[0441]

[0442] Synthesis route as Synthesis route I

[0443] The specific synthesis steps are as follows:

[0444] The carbazole in Example 1 was replaced by 5,11-dihydro-10H-dibenzo[b,f]azepine-10-one, and the rest was the same as in Example 1. The total yield was 39%.

[0445] 1 H NMR (400MHz, DMSO-d6) δ9.95 (s, 1H), 7.97-7.89 (m, 1H), 7.76-7.62 (m, 2H), 7.54-7.44 (m, 3H), 7.44-7.34 (m, 2H), 7.28 (dd, J= 7.4,1.7Hz,1H),7.20-7.05(m,2H),6.97(t,J=7.6Hz,1H),5.24(s,2H),4.01(s,2H),2.74(q,J=7.2Hz,2H),1.03-0.91(m,3H). 13 CNMR(101MHz,DMSO-d6)δ190.62,165.54,149.97,147.69,141.62,134.57,132.50,130.68,130.54, 128.98,128.57,127.74,126.29,126.10,122.80,120.99,119.45,54.91,49.32,45.97,13.52.LC-MS m / z:385.88[M+H] + .

[0446] Example 15

[0447] 4-(((2,4-Dimethyl-5H-benzo[b]pyrimidin[4,5-f]azepin-5-yl]methyl)methyl)-N'-ethylbenzohydrazide

[0448]

[0449] Synthesis route as Synthesis route I

[0450] The specific synthesis steps are as follows:

[0451] The carbazole in Example 1 was replaced by 2,4-dimethyl-5H-benzo[b]pyrimidin[4,5-f]azepine, and the rest was the same as in Example 1. The total yield was 41%.

[0452] 1 H NMR (400MHz, DMSO-d6) δ9.87(s,1H),7.63(d,J=8.1Hz,2H),7.44(d,J=8.1Hz,2H),7.24-7.16(m,1H),7.14-7.07(m,2H),7.02-6.95(m,1 H), 6.92 (d, J = 11.4Hz, 1H), 6.71 (d, J = 11.4Hz, 1H), 5.05 (s, 2H), 2.72 (q, J = 7.1Hz, 2H), 2.35 (s, 3H), 2.30 (s, 3H), 0.96 (t, J = 7.1Hz, 3H). 13 C NMR (101MHz, DMSO-d6) δ167.39,165.83,165.63,165.05,147.92,142.19,134.11,133.06,132.25,130. 03,129.77,128.30,127.46,127.32,124.85,122.29,118.79,52.43,45.98,25.62,22.37,13.60.LC-MS m / z:399.97[M+H] + .

[0453] Example 16

[0454] N'-ethyl-4-((3-(trifluoromethyl)-5H-dibenzo[b,f]azepin-5-yl)methyl)benzohydrazide

[0455]

[0456] Synthesis route as Synthesis route I

[0457] The specific synthesis steps are as follows:

[0458] The carbazole in Example 1 was replaced by 3-(trifluoromethyl)-5H-dibenzo[b,f]azepine, and the rest was the same as in Example 1. The total yield was 37%.

[0459] 1H NMR (400MHz, DMSO-d6) δ9.89 (s, 1H), 7.63 (d, J = 8.2Hz, 2H), 7.50-7.38 (m, 3H), 7.31-7.09 (m, 5H), 7 .00-6.92(m,2H),6.87(d,J=11.4Hz,1H),5.05(s,2H),2.72(q,J=7.2Hz,2H),0.96(t,J=7.1Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ165.64,151.01,150.56,142.29,139.07,134.03,132.60,132.25,131.80,130.94,1 29.35,129.26,129.20,128.19,127.47,124.61,123.86,121.70,121.09,54.05,45.99,21.35,13.59.LC-MS m / z:437.96[M+H] + .

[0460] Example 17

[0461] N'-ethyl-4-((10-hydroxy-10,11-dihydro-5H-dibenzo[b,f]azepin-5-yl)methyl)benzohydrazide

[0462]

[0463] Synthesis route as Synthesis route I

[0464] The specific synthesis steps are as follows:

[0465] The carbazole in Example 1 was replaced by 10,11-dihydro-5H-dibenzo[b,f]azepine-10-ol, and the rest was the same as in Example 1. The total yield was 37%.

[0466] 1 H NMR(400MHz,Chloroform-d)δ7.59(d,J=8.0Hz,2H),7.47-7.39(m,3H),7.22(d,J=7.4Hz,1H),7.16-7.03(m,4H),7.00-6.92(m,2H),5.20(dd,J=7 .6,3.5Hz,1H),5.08-4.96(m,2H),3.60(dd,J=13.6,3.5Hz,1H),3.24(dd,J=13.6,7.6Hz,1H),2.93(q,J=7.2Hz,2H),1.11(t,J=7.2Hz,3H).LC-MS m / z:388.14[M+H]+ .

[0467] Example 18

[0468] N'-ethyl-4-((3-methyl-5H-dibenzo[b,f]azepin-5-yl)methyl)benzohydrazide

[0469]

[0470] Synthesis route as Synthesis route I

[0471] The specific synthesis steps are as follows:

[0472] The carbazole in Example 1 was replaced by 3-methyl-5H-dibenzo[b,f]azepine, and the rest was the same as in Example 1. The total yield was 42%.

[0473] 1 H NMR(400MHz, DMSO-d6)δ9.86(s,1H),7.62(d,J=8.2Hz,2H),7.45(d,J=8.0Hz,2H),7.19-7.12(m,1H),7.11-7.02(m,2H) ,6.98-6.87(m,3H),6.78-6.71(m,3H),4.96(d,J=9.4Hz,2H),2.72(q,J=7.1Hz,2H),2.18(s,3H),0.96(t,J=7.1Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ165.64,151.01,150.56,142.29,139.07,134.03,132.60,132.25,131.80,130.94,1 29.35,129.26,129.20,128.19,127.47,124.61,123.86,121.70,121.09,54.04,45.99,21.35,13.59.LC-MS m / z:384.04[M+H] + .

[0474] Example 19

[0475] N'-ethyl-4-((3-fluoro-5H-dibenzo[b,f]azepin-5-yl)methyl)benzohydrazide

[0476]

[0477] Synthesis route as Synthesis route I

[0478] The specific synthesis steps are as follows:

[0479] The carbazole in Example 1 was replaced by 3-fluoro-5H-dibenzo[b,f]azepine, and the rest was the same as in Example 1. The total yield was 41%.

[0480] 1 H NMR(400MHz, DMSO-d6)δ9.89(s,1H),7.64(d,J=8.1Hz,2H),7.45(d,J=8.0Hz,2H),7.22-7.16(m,1H),7.13-7.04( m,3H),7.03-6.91(m,2H),6.80-6.73(m,3H),4.96(d,J=13.3Hz,2H),2.73(q,J=7.2Hz,2H),0.96(t,J=7.1Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ165.62,162.68,152.50,150.16,141.85,133.73,132.37,132.10,131.74,130.77,1 30.67,130.18,129.51,129.41,128.22,127.57,124.35,121.32,110.86,108.70,54.05,45.99,13.56.LC-MS m / z:387.88[M+H] + .

[0481] Example 20

[0482] N-(4-(2-Ethylhydrazine-1-carbonyl)benzyl)benzamide

[0483]

[0484] Synthesis route as Synthesis route III

[0485] The specific synthesis steps are as follows:

[0486] a. Synthesis of Compound 1 [Methyl 4-(Benzamidomethyl)benzoate]

[0487] Benzoic acid (500 mg, 4 mmol) was placed in a 100 mL eggplant-shaped flask at room temperature and dissolved in 30 mL of dichloromethane. 2-(7-Azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) (1.8 g, 4.8 mmol) and DIPEA (620 mg, 4.8 mmol) were added under ice and allowed to react for 0.5 hour. 4-Aminomethylbenzoic acid hydrochloride (984 mg, 4.8 mmol) was added and allowed to react overnight at room temperature. TLC confirmed the complete reaction of the starting material. The reaction mixture was then washed with water (20 mL x 3) and the organic phase was dried over anhydrous Na2SO4. Column chromatography afforded compound 1 (700 mg, 65% yield) as a white solid, which was used directly in the next step.

[0488] b. Synthesis of Compound 2 [4-(Benzamidomethyl)benzoic acid]

[0489] In step b of Example 1, 4-((9H-carbazol-9-yl)methyl)benzoic acid methyl ester was replaced with 4-(benzamidomethyl)benzoic acid methyl ester, and the rest was the same as in step b of Example 1.

[0490] c. Synthesis of Compound 3 [tert-butyl 2-(4-(benzamidomethyl)benzoyl)-1-ethylhydrazine-1-carboxylate]

[0491] In step c of Example 1, 4-((9H-carbazol-9-yl)methyl)benzoic acid was replaced with 4-(benzamidomethyl)benzoic acid, and the rest was the same as in step c of Example 1.

[0492] d. Synthesis of Example 20 [N-(4-(2-ethylhydrazine-1-carbonyl)benzyl)benzamide]

[0493] The tert-butyl 2-(4-(((9H-carbazol-9-yl)methyl)benzoyl)-1-ethylhydrazine-1-carboxylate in step d of Example 1 was replaced with tert-butyl 2-(4-(benzamidomethyl)benzoyl)-1-ethylhydrazine-1-carboxylate, and the rest was the same as step d of Example 1.

[0494] 1 H NMR (400MHz, DMSO-d6): δ10.00(s,1H),9.13-9.11(m,1H),7.93-7.90(m,2H),7.81-7.89(m,2H),7. 56-7.47(m,3H),7.42-7.39(m,2H),5.07(s,1H),4.54(s,2H),2.83-2.80(m,2H),1.06-1.02(m,3H). 13C NMR (100MHz, DMSO-d6): δ166.8,165.7,143.5,134.7,132.2,131.8,128.8,127.7,127.6,127.5,46.0,42.9,13.6.LC-MS m / z:298.25[M+H] + .

[0495] Example 21

[0496] 1-Butyl-1-(4-(2-ethylhydrazine-1-carbonyl)benzyl)-3-phenylurea

[0497]

[0498] Synthesis route as Synthesis route II

[0499] The specific synthesis steps are as follows:

[0500] a. Synthesis of Compound 1 [Methyl 4-((1-butyl-3-phenylureido)methyl)benzoate]

[0501] At room temperature, triphosgene (800 mg, 2.15 mmol) was placed in a 200 mL eggplant flask and dissolved in 50 mL of DCE. TEA (325 mg, 3.2 mmol) was then added dropwise to the mixture under ice. Aniline (500 mg, 5.3 mmol) dissolved in 5 mL of DCE was then added dropwise to the mixture. The mixture was refluxed for 4 hours. The reaction mixture was then added dropwise to methyl 4-((butylamino)methyl)benzoate (1.18 g, 5.3 mmol) dissolved in 5 mL of DCE and allowed to react at 85°C overnight. After TLC analysis, the DCE was evaporated, the mixture was dissolved in DCM, and the resulting mixture was washed with 1 M HCl (20 mL x 3). The organic phase was dried over anhydrous Na2SO4. Column chromatography afforded compound 1 (1.28 g, 71% yield) as a white solid, which was used directly in the next step.

[0502] 1 H NMR(400MHz,DMSO-d6)δ8.34(s,1H),7.95-7.84(m,2H),7.47-7.40(m,2H),7.39-7.34(m,2H),7.23-7.16(m,2H),6.96-6.86 (m,1H),4.62(s,2H),3.80(s,3H),3.33-3.24(m,2H),1.50-1.39(m,2H),1.22(q,J=14.6,7.4Hz,2H),0.82(t,J=7.3Hz,3H).

[0503] b. Synthesis of Compound 2 [4-((1-butyl-3-phenylureido)methyl)benzoic acid]

[0504] In step b of Example 1, methyl 4-((9H-carbazol-9-yl)methyl)benzoate was replaced with methyl 4-((1-butyl-3-phenylureido)methyl)benzoate, and the rest was the same as in step b of Example 1.

[0505] 1 H NMR(400MHz, DMSO-d6)δ9.48(s,1H),8.84(dd,J=4.2,1.7Hz,1H),8.47(dd,J=6.5,2.5Hz,1H),8.34-8.28(m,1H) ,8.00(t,J=5.9Hz,1H),7.92-7.85(m,2H),7.56(dd,J=8.3,4.2Hz,1H),7.51-7.36(m,4H),4.40(d,J=5.8Hz,2H).

[0506] c. Synthesis of compound 3 [tert-butyl 2-(4-((1-butyl-3-phenylureido)methyl)benzoyl)-1-ethylhydrazine-1-carboxylate].

[0507] The 4-((9H-carbazol-9-yl)methyl)benzoic acid in step c of Example 1 was replaced with 4-((1-butyl-3-phenylureido)methyl)benzoic acid, and the rest was the same as in step c of Example 1.

[0508] 1 H NMR (400MHz, DMSO-d6) δ10.44(s,1H),8.34(s,1H),7.76(d,J=9.0Hz,2H),7.46-7.40(m,2H),7.33(d,J=8.2Hz,2H),7.23-7.15(m,2H),6.94-6.87 (m,1H),4.60(s,2H),3.41(q,J=7.0Hz,2H),3.27(d,J=7.7Hz,2H),1.51- 1.34(m,6H),1.31-1.19(m,7H),1.09-0.98(m,3H),0.82(t,J=7.3Hz,3H).

[0509] d. Synthesis of Example 21 [1-butyl-1-(4-(2-ethylhydrazine-1-carbonyl)benzyl)-3-phenylurea]

[0510] The 2-(4-(((9H-carbazol-9-yl)methyl)benzoyl)-1-ethylhydrazine-1-carboxylic acid tert-butyl ester in step d of Example 1 was replaced with 2-(4-(benzamidomethyl)benzoyl)-1-ethylhydrazine-1-carboxylic acid tert-butyl ester, and the rest was the same as in step d of Example 1.

[0511] 1 H NMR(400MHz,DMSO-d6)δ9.96(s,1H),8.35(s,1H),7.80-7.74(m,2H),7.47- 7.39(m,2H),7.29(d,J=8.1Hz,2H),7.25-7.14(m,2H),6.94-6.85(m,1H),5 .08(s,1H),4.59(s,2H),3.27(t,J=7.5Hz,2H),2.82-2.71(m,2H),1.51-1. 38(m,2H),1.30-1.15(m,2H),0.99(t,J=7.2Hz,3H),0.82(t,J=7.3Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ165.68,155.78,142.96,141.01,132.41,128.76,127.74 ,127.48,122.36,120.59,49.53,46.62,46.03,30.49,19.99,14.34,13.63.LC-MS m / z:368.93[M+H] + .

[0512] Example 22

[0513] N-Butyl-N-(4-(2-ethylhydrazine-1-carbonyl)benzyl)benzamide

[0514]

[0515] Synthesis route as Synthesis route III

[0516] The specific synthesis steps are as follows:

[0517] In Example 20, 4-aminomethylbenzoic acid hydrochloride was replaced by methyl 4-((butylamino)methyl)benzoate hydrochloride, and the rest was the same as in Example 20. The total yield was 46%.

[0518] 1H NMR (400MHz, DMSO-d6) δ10.02(s,1H),7.79(d,J=8.2Hz,2H),7.39(dd,J=17.3,9.7Hz,7H),4.46(s,1H),3 .23-2.97(m,2H),2.78(q,J=7.2Hz,2H),1.52-1.20(m,4H),1.00(t,J=7.2Hz,3H),0.87(t,J=7.4Hz,3H). 13 C NMR(101MHz,Chloroform-d)δ172.36,166.95,136.41,129.65,128.61,128.18, 127.52,126.56,48.47,47.40,46.64,30.43,29.18,19.74,13.63,13.10.LC-MS m / z:335.87[M+H] + .

[0519] Example 23

[0520] 1-(4-(2-ethylhydrazine-1-carbonyl)phenyl)-3-(quinolin-8-yl)urea

[0521]

[0522] Synthesis route as Synthesis route II

[0523] The specific synthesis steps are as follows:

[0524] In Example 21, aniline and methyl 4-((butylamino)methyl)benzoate were replaced by quinolin-8-amine and methyl 4-aminobenzoate, respectively, and the rest was the same as in Example 21. The total yield was 34%.

[0525] 1 H NMR (400MHz, DMSO-d6) δ10.09(s,1H),9.87(s,1H),9.74(s,1H),8.90(dd,J=4.2,1.7Hz,1H),8.56-8.50(m,1H),8.37( dd,J=8.3,1.7Hz,1H),7.82-7.73(m,2H),7.65-7.51(m,5H),5.08(s,1H),2.77(q,J=7.1Hz,2H),1.00(t,J=7.1Hz,3H). 13C NMR(101MHz,DMSO-d6)δ165.56,152.74,148.91,143.16,138.27,137.20,136.18,12 8.62,128.46,127.76,126.83,122.63,120.60,117.64,115.07,46.12,13.65.LC-MS m / z:350.03[M+H] + .

[0526] Example 24

[0527] 1-(4-(2-ethylhydrazine-1-carbonyl)benzyl)-3-(quinolin-8-yl)urea

[0528]

[0529] Synthesis route as Synthesis route II

[0530] The specific synthesis steps are as follows:

[0531] The methyl 4-aminobenzoate in Example 23 was replaced by methyl 4-aminomethylbenzoate hydrochloride, and the rest was the same as in Example 23. The total yield was 42%.

[0532] 1 H NMR (400MHz, DMSO-d6) δ9.96 (s, 1H), 9.46 (s, 1H), 8.84 (dd, J = 4.2, 1.7Hz, 1H), 8.4 8(dd,J=6.9,2.2Hz,1H),8.31(dd,J=8.3,1.7Hz,1H),7.95(t,J=5.9Hz,1H),7.82- 7.72(m,2H),7.56(dd,J=8.2,4.1Hz,1H),7.50-7.42(m,2H),7.37(d,J=8.0Hz,2H) ,5.03(s,1H),4.38(d,J=5.8Hz,2H),2.77(q,J=7.2Hz,2H),0.99(t,J=7.1Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ165.72,155.72,148.63,144.13,138.12,137.06,137.01,132.2 7,128.43,127.75,127.69,127.43,122.42,119.66,114.51,46.03,43.00,13.64.LC-MS m / z:363.97[M+H] + .

[0533] Example 25

[0534] N'-ethyl-4-((5-(thiophen-2-yl)-1H-tetrazol-1-yl)methyl)benzohydrazide

[0535]

[0536] Synthesis route as Synthesis route I

[0537] The specific synthesis steps are as follows:

[0538] The carbazole in Example 1 was replaced by 5-(thiophen-2-yl)-1H-tetrazole, and the rest was the same as in Example 1. The total yield was 35%.

[0539] 1 H NMR (400MHz, DMSO-d6) δ10.03 (s, 1H), 7.88-7.68 (m, 4H), 7.43 (d, J = 8.1Hz, 2H), 7.26 -7.14(m,1H),6.01(s,2H),5.08(s,1H),2.76(q,J=7.2Hz,2H),0.98(t,J=7.2Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ165.36,161.17,137.46,134.03,129.80,129.03,128.82,128.60,128.18,56.19,56.14,45.97,13.61.LC-MS m / z:328.93[M+H] + .

[0540] Example 26

[0541] N'-ethyl-2-(pyridin-3-yl)thiazole-4-carbohydrazide

[0542]

[0543] The specific synthesis steps are as follows:

[0544] a. Synthesis of Compound 1 [2-(pyridin-3-yl)thiazole-4-carboxylic acid]

[0545] In step b of Example 1, methyl 4-((9H-carbazol-9-yl)methyl)benzoate was replaced with methyl 2-(pyridin-3-yl)thiazole-4-carboxylate, and the rest was the same as in step c of Example 1.

[0546] b. Synthesis of Compound 2 [tert-butyl 1-ethyl-2-(2-(pyridin-3-yl)thiazole-4-carbonyl)hydrazine-1-carboxylate]

[0547] In step c of Example 1, 4-((9H-carbazol-9-yl)methyl)benzoic acid was replaced with compound 1. The rest was the same as in step c of Example 1. The yield was 56%.

[0548] 1 H NMR (400MHz, DMSO-d6) δ10.61 (s, 1H), 9.32-9.22 (m, 1H), 8.68 (dd, J = 4.8, 1.6Hz, 1H), 8.47 (s, 1H) ,8.44-8.38(m,1H),7.59-7.52(m,1H),3.45(q,J=7.2Hz,2H),1.36(s,9H),1.13(t,J=7.1Hz,3H).

[0549] c. Synthesis of Compound 3 [N'-ethyl-2-(pyridin-3-yl)thiazole-4-carbohydrazide]

[0550] In step d of Example 1, tert-butyl 2-(4-(((9H-carbazol-9-yl)methyl)benzoyl)-1-ethylhydrazine-1-carboxylate was replaced with compound 2. The rest was the same as in step d of Example 1. The yield was 78%. 1 H NMR (400MHz, DMSO-d6) δ10.06 (s, 1H), 9.26-9.19 (m, 1H), 8.64 (dd, J = 4.8, 1.6Hz, 1H), 8.39 -8.32(m,2H),7.54-7.47(m,1H),5.10(s,1H),2.83(q,J=7.2Hz,2H),1.00(t,J=7.2Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ164.78,159.81,151.78,150.29,147.84,134.45,128.98,125.12,124.67,46.03,13.58.LC-MS m / z:248.92[M+H] + .

[0551] Example 27

[0552] N'-Ethyl-4-((quinolin-8-ylamino)methyl)benzohydrazide

[0553]

[0554] Synthesis route as Synthesis route I

[0555] The specific synthesis steps are as follows:

[0556] The carbazole in Example 1 was replaced by quinolin-8-amine, and the rest was the same as in Example 1. The total yield was 35%.

[0557] 1 H NMR(400MHz, DMSO-d6)δ9.93(s,1H),8.73(dd,J=4.0,1.8Hz,1H),8.21-8.12(m,1H),7.78-7.70(m,2H),7.52-7.38(m,3H),7.29-7.15 (m,2H),7.01(d,J=8.1Hz,1H),6.47(d,J=7.7Hz,1H),5.16(s,1H),4.56(d,J=6.3Hz,2H),2.76(q,J=7.3Hz,2H),0.98(t,J=7.3Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ165.79,147.53,144.67,144.02,138.12,136.52,132.30,1 28.85,128.16,127.72,127.37,122.29,114.00,105.42,46.39,46.03,13.62.LC-MS m / z:320.96[M+H] + .

[0558] Example 28

[0559] 4-((2,4-dioxy-3-phenethyl-3,4-dihydroquinazolin-1(2H)-yl)methyl)-N'-ethylbenzohydrazide

[0560]

[0561] Synthesis route as Synthesis route I

[0562] The specific synthesis steps are as follows:

[0563] The carbazole in Example 1 was replaced by N'-ethylbenzoylhydrazide-3-phenylethylquinazoline-2,4(1H,3H)-dione, and the rest was the same as in Example 1. The total yield was 31%.

[0564] 1 H NMR(400MHz, DMSO-d6)δ9.97(s,1H),8.04(dd,J=7.8,1.6Hz,1H),7.77-7.70(m,2H),7.64-7.57(m,1H),7.31-7.14( m,9H),5.36(s,2H),5.04(s,1H),4.29-4.11(m,2H),3.02-2.87(m,2H),2.76(q,J=7.2Hz,2H),0.98(t,J=7.1Hz,3H). 13C NMR(101MHz,DMSO-d6)δ165.51,161.48,151.20,140.14,140.00,139.06,135.77,132.77,129.30,128.98,1 28.57,127.95,126.92,123.53,115.68,115.40,46.60,45.99,43.03,33.66,13.62.LC-MSm / z:443.14[M+H] + .

[0565] Example 29

[0566] N'-ethyl-4-((5-(4-methoxybenzoyl)-1H-indol-1-yl)methyl)benzohydrazide

[0567]

[0568] Synthesis route as Synthesis route I

[0569] The specific synthesis steps are as follows:

[0570] The carbazole in Example 1 was replaced by N'-ethylbenzoylhydrazide-(1H-indol-5-yl)(4-methoxyphenyl)methanone, and the rest was the same as in Example 1. The total yield was 41%.

[0571] 1 H NMR(400MHz, DMSO-d6)δ9.96(s,1H),7.95(d,J=1.8Hz,1H),7.77-7.67(m,4H),7.64(dd,J=3.2,1.0Hz,1H),7.58-7.49(m,2H),7.25(d,J=8.1 Hz,2H),7.07-7.00(m,2H),6.67-6.63(m,1H),5.52(s,2H),5.12(s,1H),3.81(d,J=1.0Hz,3H),2.75(q,J=7.1Hz,2H),0.97(t,J=7.2Hz,3H). 13 C NMR (101MHz, DMSO-d6) δ195.22,165.53,162.84,141.60,138.30,133.01,132.73,132.47,131.58,131.23,129. 75,128.11,127.95,127.47,124.60,123.49,114.38,114.18,110.66,103.62,55.99,49.50,45.99,13.60.LC-MS m / z:427.93[M+H] +.

[0572] Example 30

[0573] 4-((4',5'-dihydrospiro[piperidin-4,7'-thienyl[2,3-c]pyran]-1-yl)methyl)-N'-ethylbenzohydrazide

[0574]

[0575] Synthesis route as Synthesis route I

[0576] The specific synthesis steps are as follows:

[0577] The carbazole in Example 1 was replaced by N'-ethylbenzoylhydrazide-4',5'-dihydrospiro[piperidin-4,7'-thieno[2,3-c]pyran], and the rest was the same as in Example 1. The total yield was 34%.

[0578] 1 H NMR (400MHz, DMSO-d6) δ10.07(s,1H),7.83(d,J=7.7Hz,2H),7.66(s,2H),7.37(d,J=5.0Hz,1H),6.80(d,J=5.0Hz,1H),4.35(s,1H),3.82(t,J=5. 5Hz,2H),3.12(d,J=4.7Hz,2H),2.82-2.73(m,2H),2.59(t,J=5.4Hz,2H) ,2.46(s,2H),2.25(d,J=25.2Hz,1H),2.06(s,2H),0.99(t,J=7.1Hz,3H). 13 C NMR(101MHz,DMSO-d6)δ171.45,165.65,137.18,129.80,128.99,127.87,12 6.84,65.56,55.46,48.71,47.35,46.01,30.53,19.63,19.19,13.58.LC-MS m / z:368.93[M+H] + .

[0579] Example 31

[0580] 2-(4-Bromophenyl)-N'-ethyloxazole-4-carbohydrazide

[0581]

[0582] The specific synthesis steps are as follows:

[0583] In Example 26, methyl 2-(pyridin-3-yl)thiazole-4-carboxylate was replaced by methyl 2-(4-bromophenyl)oxazole-4-carboxylate, and the rest was the same as in Example 26. The total yield was 36%.

[0584] 1 H NMR (400MHz, DMSO-d6) δ9.84(s,1H),8.68(s,1H),7.92-7.87(m,2H),7.78-7.69(m,2H),5.04(s,1H),2.78(q,J=7.2Hz,2H),0.98(t,J=7.2Hz,3H). 13 CNMR(101MHz,DMSO-d6)δ160.37,159.19,142.64,136.89,132.86,128.72,125.96,125.30,45.94,13.53.LC-MSm / z:386.03[M+H] + .

[0585] Example 32

[0586] 8-(2-Ethylhydrazinyl)-8-oxo-N-phenyloctanamide

[0587]

[0588] The specific synthesis steps are as follows:

[0589] In Example 26, methyl 2-(pyridin-3-yl)thiazole-4-carboxylate was replaced by methyl 8-oxo-8-(phenylamino)octanoate, and the rest of the reaction was the same as in Example 26. The total yield was 46%.

[0590] 1 H NMR(400MHz,DMSO-d6)δ9.82(s,1H),9.19(s,1H),7.59-7.49(m,2H),7.28-7.18(m,2H),7.02-6.93(m,1H),4.76(s,1H),2. 68-2.55(m,2H),2.24(t,J=7.4Hz,2H),1.97(t,J=7.4Hz,2H),1.59-1.38(m,4H),1.31-1.16(m,4H),0.91(t,J=7.2Hz,3H). 13 C NMR (101MHz, DMSO-d6) δ171.76,171.47,139.89,129.18,123.44,119.53,45.93,36.89,33.99,28.93,25.66,25.56,13.57.LC-MS m / z:292.02[M+H]+ .

[0591] Example 33

[0592] N-(4-(2-Methylhydrazine-1-carbonyl)benzyl)benzamide

[0593]

[0594] Synthesis route as Synthesis route III

[0595] The specific synthesis steps are as follows:

[0596] In Example 20, tert-butyl 1-ethylhydrazine-1-carboxylate was replaced with tert-butyl 1-methylhydrazine-1-carboxylate, and the rest was the same as in Example 20. The total yield was 41%.

[0597] 1 H NMR (400MHz, DMSO-d6) δ12.01(s,1H),9.19(t,J=6.0Hz,1H),7.94-7.82(m,4H),7.55-7.39(m,5H),4.52(d,J=6.0Hz,2H),2.83(s,3H). 13 C NMR(101MHz,DMSO-d6)δ166.86,165.75,145.71,134.64,131.92,129.11,128.90,128.50,127.89,127.83,39.70.LC-MS m / z:283.92[M+H] + .

[0598] Example 34

[0599] 8-(2-Methylhydrazinyl)-8-oxo-N-phenyloctanamide

[0600]

[0601] The specific synthesis steps are as follows:

[0602] In Example 32, tert-butyl 1-ethylhydrazine-1-carboxylate was replaced with tert-butyl 1-methylhydrazine-1-carboxylate, and the rest was the same as in Example 20. The total yield was 41%.

[0603] 1H NMR(400MHz,DMSO-d6)δ9.87(s,1H),9.21(s,1H),7.56-7.50(m,2H),7.27-7.16(m,2H),7.07-7.01(m,1H) ,4.58(s,1H),2.47(s,3H),2.41-2.32(m,2H),1.97(t,J=7.4Hz,2H),1.59-1.38(m,4H),1.31-1.16(m,4H). 13 C NMR(101MHz,DMSO-d6)δ172.56,171.74,139.68,128.34,123.64,120.65,36.78,34.07,28.61,25.58,25.49,13.42.LC-MS m / z:278.06[M+H] + .

[0604] Test Example 1 In vitro inhibitory activity of HDAC1, 2, 3, 6

[0605] 50 μL of drug-containing HDAC buffer was mixed with 10 μL of enzyme solution and pre-incubated for 5 minutes. After adding 40 μL of substrate, the reaction was carried out at 37°C for 30 minutes. Then, 100 μL of trypsin stop solution was added to terminate the reaction. The reaction was continued at 37°C for 20 minutes, and the fluorescence intensity was measured at 390 nm / 460 nm.

[0606]

[0607] Finally, the inhibition rate (%) of the compound and its corresponding concentration were fitted with an S curve to calculate the IC50 value.

[0608] The inhibitory activity results are shown in Table 1 below. The experimental results show that most of the above compounds have nanomolar inhibitory IC against HDAC6. 50 The activity against HDAC1, 2 and 3 is generally lower than that against HDAC6. Among them, Examples 1, 9, 12, 13, 17 and 27 have a selectivity of 10-30 times against class I HDAC, and their selectivity and activity are significantly better than the positive control drug Tubastatin A.

[0609] Table 1: Examples and IC values ​​of Tubastatin A for HDAC1, 2, 3, and 6 50 value

[0610]

[0611]

[0612] Experimental Example 2: Selectivity for HDAC6 Cell Levels

[0613] MV4-11 and JA74.1 cells were seeded in 6-well cell culture plates (1×106 cells / well). After 12 hours, the cells were treated with gradient concentrations of the compounds for 24 hours. After 24 hours, the cells were collected, washed twice with PBS, and total protein was extracted with cold RIPA lysis buffer (50mM Tris base, 150mM NaCl, 5mM EDTA, 0.1% (v / v) SDS, 0.5% (v / v) sodium deoxycholate, and 1% (v / v) Triton-x-100). The suspension was centrifuged at 12,000 rpm for 15 minutes at 4°C, and the protein concentration at different compound doses was detected using a bicinchoninic acid (BCA) protein assay. 80 μL of the supernatant was mixed with 20 μL of β-mercaptoethanol and NuPAGE lithium dodecyl sulfate (LDS) sample buffer (5X). The mixture was heated at 100°C for 10 minutes to obtain the loading sample. Equal amounts of protein were separated by 12% SDS-polyacrylamide gel electrophoresis at 120 V and transferred to polyvinylidene difluoride membranes at 250 mA at 4°C. After blocking with a solution containing 0.05% Tween-20 (TBST) and 5% skim milk for 2 hours at room temperature, the membranes were incubated with primary antibodies overnight at 4°C in TBS buffer containing 0.05% Tween-20 (TBST), followed by incubation with horseradish (HRP)-conjugated secondary antibodies at a dilution of 1:5000.

[0614] The experimental results are as follows Figure 1 As shown in the table, Example 13 still has high selectivity at 2500 nM compared to the positive compound SAHA in MV4-11 and J774A.1 cells. Therefore, Example 13 is a potential HDAC6 selective inhibitor.

[0615] Experimental Example 3 Time dependence of HDAC6 inhibition

[0616] 50 μL of drug-containing HDAC buffer was mixed with 10 μL of enzyme solution and pre-incubated for 0 min, 15 min, 30 min, and 1 h. Afterwards, 40 μL of substrate was added and the reaction was incubated at 37°C for 30 min. The reaction was terminated by adding 100 μL of trypsin stop solution and incubated at 37°C for 20 min. Fluorescence intensity was measured at 390 nm / 460 nm. The deacetylation activity (%) was calculated based on the fluorescence intensity, and curve fitting was performed using the "log (inhibitor) vs. normalized response-variable slope (four parameters)" function in GraphPad Prism software.

[0617] The experimental results are as follows Figure 2 As shown, by comparing the IC values ​​of Examples 3, 13 and 27 and the positive control Tubastatin A at 0 min, 15 min, 30 min and 1 h 50 We found that the inhibitory activity of Examples 3, 13, and 27, which use hydrazide as a chelating group, against HDAC6 gradually increased with prolonged incubation time with the HDAC6 enzyme, indicating that they bind to HDAC6 in a slow binding manner. However, the inhibitory activity of the hydroxamic acid Tubastatin A against HDAC6 did not change significantly with prolonged incubation time with the HDAC6 enzyme, indicating that its binding mode with HDAC6 is fast binding. This concludes that HDAC6 inhibitors using hydrazide and hydroxamic acid as chelating groups have different binding modes with HDAC6.

[0618] Test Example 4 Rapid Dilution Experiment

[0619] 50 μg / mL HDAC6 was incubated with 1 μM and 10 nM of Examples 3, 13, and 27, and 2 μM and 20 nM Tubastatin A, respectively, in HDAC buffer for 1 h. 1 μL of this mixture was diluted to 100 μL with 1 μM and 10 nM compound solutions, respectively. This solution also contained 50 μM HDAC substrate and 2 milliunits of Lys-C protease solution. Fluorescence intensity was then measured at 360 nm (ex.) / 460 nm (em.) every 2 minutes for one hour.

[0620] The experimental results are as follows Figure 3 As shown, the complexes formed by Examples 3, 13, and 27 with HDAC6 are more stable and maintain the same inhibitory activity after 100-fold dilution. However, the complex formed by the hydroxamic acid Tubastatin A with HDAC6 is unstable after 100-fold dilution and fails to maintain its inhibitory activity against HDAC6. The conclusion is that Examples 3, 13, and 27 dissociate slowly from HDAC6, indicating that they are long-acting HDAC6 inhibitors, while the hydroxamic acid HDAC6 inhibitor Tubastatin A dissociates rapidly from HDAC6, indicating that it does not bind stably to HDAC6.

[0621] Test Example 5 Anti-inflammatory activity test

[0622] J774A.1 cells were seeded into 96-well plates (1×105 cells / well) and cultured in growth medium for 24 h. The cells were primed with E. coli 0111:B4 LPS (Sigma-Aldrich) (final concentration: 1 μg / mL) for 4.5 h. Next, test compounds (0.1, 0.3, 1.0, 3.0, and 10.0 μM) were added over 30 min. ATP (5 mM) was added simultaneously with the addition of the compounds to induce NLRP3 inflammasome activation. After 30 min, the supernatant was collected and IL-1β levels were measured using a mouse IL-1β ELISA kit (DuoSet ELISA, R&D Systems) according to the manufacturer's instructions.

[0623] The test results are as follows Figure 4 As shown in Figure 2, in J774A.1 cells, Example 13 was able to significantly reduce the level of IL-1β induced by LPS / ATP in a concentration-dependent manner, with an IC 50 The results showed that the concentration of 1 μg / mL of PEG-1β-catenin was 2.61±0.24 μM, demonstrating its inhibitory activity against NLRP3.

[0624] The foregoing merely illustrates the principles of the present invention. It should be understood that the scope of the present invention is not intended to be limited to the exemplary aspects described herein, but rather includes all currently known and future developed equivalents. In addition, it should be noted that various improvements and modifications may be made without departing from the technical principles of the present invention, and such improvements and modifications should also be considered within the scope of the present invention.

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof, characterized in that Its structure is as follows: in Z is a key; Where Y is When: R1 and R6 are each independently hydrogen; Y is selected from (C3-C5) alkynyl, When: R6 is selected from H, optionally substituted (C1-C4) alkyl, R1 is selected from H, methyl; Wherein, R is selected from or benzene ring, R is optionally substituted by 1-3 substituents selected from halogen, halo (C1-C 12 )alkyl, (C1-C 12 )alkyl, (C2-C 12 )alkenyl, (C3-C5)alkynyl, (C1-C2)alkoxy, hydroxy, phenolic hydroxy, cyano or (C1-C2)formate; W is selected from C or N, Ring A is selected from a 5-6 membered aliphatic ring, aliphatic heterocyclic ring, aromatic ring or heteroaromatic ring, Ring B is selected from a 5-6 membered aliphatic ring, aliphatic heterocyclic ring, aromatic ring or heteroaromatic ring, Ring C is selected from H, 5-6 membered aromatic ring or aromatic heterocyclic ring, X is selected from CH2, O, S, (C=O), 2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Ring A is optionally substituted by one or more substituents, wherein the substituents are selected from halogen, halo (C1-C 12 )alkyl, (C1-C 12 )alkyl, (C2-C 12 )alkenyl, (C3-C5)alkynyl, (C1-C2)alkoxy, phenolic hydroxyl, cyano or (C1-C2)formate.

3. The compound according to claim 2 or a pharmaceutically acceptable salt thereof, wherein Ring A is selected from one or more of the following ring systems:

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein Ring B is optionally substituted with one or more substituents, wherein the substituents are selected from halogen, halo (C1-C 12 )alkyl, (C1-C 12 )alkyl, (C2-C 12 )alkenyl, (C3-C5)alkynyl, (C1-C2)alkoxy, phenolic hydroxyl, cyano or (C1-C2)formate.

5. The compound according to claim 4 or a pharmaceutically acceptable salt thereof, wherein Ring B is selected from one or more of the following ring systems:

6. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein The C ring is optionally substituted by one or more substituents, wherein the substituents are selected from halogen, halo (C1-C 12 )alkyl, (C1-C 12 )alkyl, (C2-C 12 )alkenyl, (C3-C5)alkynyl, (C1-C2)alkoxy, phenolic hydroxyl, cyano or (C1-C2)formate.

7. The compound according to claim 6 or a pharmaceutically acceptable salt thereof, wherein Ring C is selected from one or more of the following ring systems:

8. The compound according to claim 7 or a pharmaceutically acceptable salt thereof, wherein Ring C is pyridine.

9. A compound or a pharmaceutically acceptable salt thereof, which is: 4-((9H-carbazol-9-yl)methyl)-N'-ethylbenzohydrazide; N'-ethyl-4-((1,2,3,4-tetrahydro-9H-carbazol-9-yl)methyl)benzohydrazide; 4-((9H-pyrido[2,3-b]indol-9-yl)methyl)-N'-ethylbenzohydrazide; 4-((9H-pyrido[3,4-b]indol-9-yl)methyl)-N'-ethylbenzohydrazide; 4-((5H-pyrido[4,3-b]indol-5-yl)methyl)-N'-ethylbenzohydrazide; 4-(Acridin-10(9H)-ylmethyl)-N'-ethylbenzohydrazide; 4-((10H-Benzoxazin-10-yl)methyl)-N'-ethylbenzohydrazide; 4-((10H-phenothiazin-10-yl)methyl)-N'-ethylbenzohydrazide; 4-((10H-benzo[b]pyrido[2,3-e][1,4]thiazin-10-yl)methyl)-N'-ethylbenzohydrazide; 4-((10H-benzo[b]pyrido[2,3-e][1,4]thiazin-10-yl)methyl)-N'-methylbenzohydrazide; 4-((10,11-dihydro-5H-dibenzo[b,f]azepin-5-yl)methyl)-N'-ethylbenzohydrazide; 4-((5H-dibenzo[b,f]azepin-5-yl)methyl)-N'-ethylbenzohydrazide; 4-((11H-benzo[b]pyrido[4,3-f]heptidin-11yl)methyl)-N'-ethylbenzohydrazide; N'-ethyl-4-((10-oxo-10,11-dihydro-5H-dibenzo[b,f]heptidin-5-yl)methyl)benzohydrazide; 4-(((2,4-Dimethyl-5H-benzo[b]pyrimidin[4,5-f]azepin-5-yl]methyl)methyl)-N'-ethylbenzohydrazide; N'-ethyl-4-((3-(trifluoromethyl)-5H-dibenzo[b,f]azepin-5-yl)methyl)benzohydrazide; N'-ethyl-4-((10-hydroxy-10,11-dihydro-5H-dibenzo[b,f]azepin-5-yl)methyl)benzohydrazide; N'-ethyl-4-((3-methyl-5H-dibenzo[b,f]azepin-5-yl)methyl)benzohydrazide; N'-ethyl-4-((3-fluoro-5H-dibenzo[b,f]azepin-5-yl)methyl)benzohydrazide; N'-ethyl-4-((5-(thiophen-2-yl)-1H-tetrazol-1-yl)methyl)benzohydrazide; N'-ethyl-4-((quinolin-8-ylamino)methyl)benzohydrazide; 4-((2,4-dioxy-3-phenethyl-3,4-dihydroquinazolin-1(2H)-yl)methyl)-N'-ethylbenzohydrazide; 4-((4',5'-dihydrospiro[piperidin-4,7'-thieno[2,3-c]pyran]-1-yl)methyl)-N'-ethylbenzohydrazide; or N-(4-(2-methylhydrazine-1-carbonyl)benzyl)benzamide.

10. A method for preparing the compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 8, comprising the following steps:

11. The preparation method according to claim 10, wherein The intermediate 1 can be obtained by the following method: using a polycyclic compound containing an aromatic secondary amine as a raw material, reacting it with methyl 4-(bromomethyl)benzoate under the action of NaH to obtain the intermediate 1; or The intermediate 1 can be obtained by the following method: using a compound containing an aromatic amino group as a raw material, reacting it with triphosgene under alkaline conditions to generate an isocyanate intermediate, and then reacting it with substituted or unsubstituted 4-aminomethylbenzoic acid methyl ester hydrochloride or 4-aminobenzoic acid methyl ester to obtain the intermediate 1; or The intermediate 1 can be obtained by the following method: using an aryl formic acid compound as a raw material, condensing it with substituted or unsubstituted 4-aminomethylbenzoic acid methyl ester hydrochloride or 4-aminobenzoic acid methyl ester under the action of HATU to obtain the intermediate 1.

12. A pharmaceutical composition comprising the compound according to any one of claims 1 to 9 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable diluent or carrier.

13. Use of the compound according to any one of claims 1 to 9 or a pharmaceutically acceptable salt thereof in the preparation of an HDAC6 inhibitor or in the preparation of a medicament for a disease associated with abnormal expression of HDAC6 activity. 14 . The use according to claim 13 , wherein the disease associated with abnormal expression of HDAC6 activity comprises tumors, neurodegenerative diseases, non-neurodegenerative nervous system diseases, inflammation or non-inflammatory autoimmune diseases.

15. The use according to claim 14, wherein The tumors include triple-negative breast cancer, lung cancer, melanoma, esophageal cancer, prostate cancer, breast cancer, cervical cancer, ovarian cancer, gastric cancer, pancreatic cancer, bladder cancer, colorectal cancer, brain tumors, bone cancer, and soft tissue sarcoma; or the neurodegenerative diseases include Parkinson's disease (PD), Alzheimer's disease (AD), cerebral ischemia (CI), brain injury (BI), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS), different types of spinocerebellar ataxia (SCA), epilepsy, and Pick's disease.

16. The use according to claim 15, wherein The brain tumor comprises glioma; the glioma comprises anaplastic oligodendroglioma, adult glioblastoma erythematosus and adult anaplastic astrocytoma.

Images