2-cyano-2-(9h-xanthene-9-ylidene)acetamides, processes for their preparation, pharmaceutical compositions and uses thereof

Abstract

The application discloses a 2-cyano-2-(9H-xanthen-9-ylidene)acetamide compound and a preparation method, a pharmaceutical composition and an application thereof. The compound structure is as shown in the formula (I), and the compound also comprises pharmaceutically acceptable salts thereof. The compound can effectively inhibit a target point ZNF207 and can be used for treating various tumors. In addition, the preparation method disclosed by the application is simple and easy to operate.

Description

TECHNICAL FIELD

[0001] The present application relates to a kind of 2-cyano-2-(9H-xanthene-9-ylidene)acetamides and its preparation method, pharmaceutical composition and application, especially to a kind of 2-cyano-2-(9H-xanthene-9-ylidene)acetamides with antitumor activity and its preparation method, pharmaceutical composition and application. BACKGROUND

[0002] Zinc finger proteins (ZFPs) are the largest class of DNA / RNA binding proteins in eukaryotes, named for the "finger-like" domains stabilized by zinc ions (Zn² + ) in their structures. Since first discovered in the transcription factor TFIIIA of Xenopus laevis in 1985, the zinc finger protein family has been proven to be widely involved in key biological processes such as gene expression regulation, cell differentiation, signal transduction, epigenetic modification, etc. The family has numerous members, and more than 800 zinc finger protein genes have been identified in the human genome, accounting for about 3% of all coding genes, showing high functional diversity.

[0003] Zinc finger protein 207 (ZNF207, also known as BuGZ) is an important member of the zinc finger family, which is a nuclear protein involved in mitotic regulation and transcriptional regulation and other cellular functions. In structure, ZNF207 mainly contains two functional domains: C2H2-type zinc finger domain (located at C-terminal) and GLEBS domain (located at N-terminal). Among them, the zinc finger domain usually has DNA or RNA binding ability, and may be involved in gene transcription regulation, although its specific target gene has not been completely clear. The GLEBS domain is a conserved protein-protein interaction sequence, which can specifically bind to mitotic checkpoint protein Bub3, thereby mediating the recruitment of Bub1 on kinetochore, which is a key step to ensure the correct separation of chromosomes in the spindle assembly checkpoint (SAC). ZNF207 also has liquid-liquid phase separation properties, which can form dynamic biomacromolecular condensates, stabilize Bub3 and promote its localization on kinetochore. The deletion of ZNF207 will destroy the formation of Bub3-Bub1 complex, leading to chromosome misalignment, mitotic elongation and apoptosis. In addition to the mitotic-related functions, ZNF207 also plays an important role in stem cell maintenance and tumorigenesis, and shows pro-cancer effect in various solid tumors such as human breast cancer, lung adenocarcinoma, hepatocellular carcinoma, esophageal cancer, etc. Among them, ZNF207 is significantly highly expressed in high-grade glioma such as glioblastoma (GBM), and its expression level is negatively correlated with the prognosis of patients. Functionally, ZNF207 can maintain the self-renewal ability of glioma stem-like cells (GSCs), enhance their anti-apoptotic properties and resistance to radiotherapy and chemotherapy. Knockout of ZNF207 can significantly reduce the expression of stemness markers such as SOX2 and Nestin in GSCs, inhibit neurosphere formation, and enhance the sensitivity to temozolomide (TMZ) and radiotherapy. Based on these findings, ZNF207 has been identified as a highly valuable molecular target for research.

[0004] The development of small molecule inhibitors targeting ZNF207 is expected to become a new strategy for the treatment of malignant tumors, which has important clinical application prospects and research value. SUMMARY

[0005] The first object of the present application is to provide a 2-cyano-2-(9 H xanthen-9-ylidene)acetamide compound, the second object is to provide a preparation method of the compound, the third object is to provide a pharmaceutical composition comprising the compound, and the fourth object is to provide the application of the compound and the pharmaceutical composition thereof.

[0006] Technical scheme: the 2-cyano-2-(9 HXanthene-9-ylidene)acetamides have the structure of Formula (I), which also includes isomers, prodrugs, stable isotopically substituted compounds, pharmaceutically acceptable salts, or mixtures thereof: wherein:

[0007] A ring is selected from ;

[0008] X is selected from ;

[0009] n is a positive integer from 0 to 2;

[0010] R 1 is selected from substituted C6~ C 10 aryl, substituted C5~ C9 heteroaryl, substituted C3~ C8 cycloalkyl, substituted C2~C6 heterocycloalkyl, substituted C4~ C 12 spirocyclyl, or substituted C4~ C 12 bridged cyclyl, wherein the substituents are selected from at least one hydrogen, halogen, cyano, phenyl, hydroxyl, amino, methylamino, dimethylamino, Boc-amino, acetylamino, sulfonamido, carboxyl, methoxycarbonyl, nitro, C1~ C3 alkyl, C1~ C3 haloalkyl, C1~ C3 alkoxy, C1~ C3 fluoroalkoxy, C2~ C6 heterocycloalkyl, benzyloxy, p-toluenesulfonate, cyclohexylmethoxy, tetrahydropyranylmethoxy, morpholinoethoxy, adamantylethanoate, phenoxy methyl, N- Boc-piperidinomethoxy, benzylamino, dimethylaminoethoxy, pyrimidin-2-ylmethoxy, phenethoxy, 2-fluorophenethoxy, pyridin-4-ylmethoxy, 4-(trifluoromethoxy)benzyloxy, 4-fluorobenzyloxy, 2-(pyridin-2-yl)ethoxy, 2-phenoxyethoxy, 2-bromophenethoxy, 4-(difluoromethoxy)benzyloxy, cyclopropylmethoxy, 2-(piperidin-1-yl)ethoxy, 2-(pyrrolidin-1-yl)ethoxy, (4-methoxyphenyl)-2-oxoethoxy, the heteroaryl, heterocycloalkyl groups comprising 1 to 3 heteroatoms selected from N, O, S.

[0011] Further,

[0012] A ring is selected from ;

[0013] X is selected from ;

[0014] n is a positive integer from 0 to 2; R 1 is selected from substituted phenyl ring, substituted naphthalene ring, substituted quinoline, substituted benz[ d ][1,3]dioxane, substituted cyclohexane, or substituted benzofuran, substituted benz[ doxazole, substituted quinoxaline, substituted 2,3-dihydrobenzo[ b ][1,4]dioxane, substituted 1 H indole, wherein the substituents are selected from at least one hydrogen, halogen, cyano, phenyl, hydroxy, amino, Boc-amino, sulfonamido, methoxycarbonyl, nitro, methyl, methoxy, trifluoromethoxy, difluoromethoxy, benzyloxy, p-toluenesulfonate, cyclohexylmethoxy, tetrahydropyranylmethoxy, morpholinoethoxy, adamantaneacetate, phenoxy methyl, N- Boc-piperidinomethoxy, benzylamino, dimethylaminoethoxy, pyrimidin-2-ylmethoxy, phenethoxy, 2-fluorophenethoxy, pyridin-4-ylmethoxy, 4- (trifluoromethoxy)benzyloxy, 4-fluorobenzyloxy, 2-(pyridin-2-yl)ethoxy, 2- phenoxyethoxy, 2-bromophenethoxy, 4-(difluoromethoxy)benzyloxy, cyclopropylmethoxy, 2-(piperidin-l-yl)ethoxy, 2-(pyrrolidin-l-yl)ethoxy, (4- methoxyphenyl)-2-oxoethoxy.

[0015] Further,

[0016] A ring is selected from ;

[0017] X is selected from ;

[0018] R 1 is selected from , substituted or unsubstituted C3~ C 10 cycloalkyl, 6-10 membered aromatic heterocycle containing 1-2 N atoms, substituted or unsubstituted benzo 5-6 membered heterocycle;

[0019] Ring B is selected from: substituted or unsubstituted C6~ C 10 aryl;

[0020] R 2 is selected from: hydrogen, nitro, cyano, halogen, trifluoromethoxy, difluoromethoxy, sulfonamido, methoxycarbonyl, methoxy, hydroxy, methyl, amino, Boc-protected amino, trifluoromethyl, phenyl;

[0021] Z is selected from: substituted or unsubstituted C6~ C 10 aryl, 6-10 membered aromatic heterocycle containing 1-2 N atoms, C3~ C 10 cycloalkyl, saturated heterocycle containing N atoms and / or O atoms, dimethylamino;

[0022] Y is selected from: O, none;

[0023] n is any one integer from 0, 1 or 2.

[0024] Further,

[0025] A ring is selected from ;

[0026] X is selected from ;

[0027] R 1 is selected from substituted or unsubstituted C3~ C 10 cycloalkyl, substituted or unsubstituted benzene 5-6 membered heterocycle;

[0028] Ring B is selected from: substituted or unsubstituted C6~ C 10 aryl;

[0029] R 2 is selected from: hydrogen, halogen, trifluoromethoxy, difluoromethoxy, sulfonamide group, methoxycarbonyl, methoxy, hydroxyl, methyl, amino, Boc protected amino, phenyl;

[0030] Z is selected from: substituted or unsubstituted C6~ C 10 aryl, 6-10 membered aromatic heterocycle containing 1-2 N atoms, C3~ C 10 cycloalkyl, saturated heterocycle containing N atom and / or O atom, dimethylamino;

[0031] Y is selected from: O, none;

[0032] n is any one integer from 0, 1 or 2.

[0033] Further,

[0034] Ring B is benzene ring;

[0035] R 2 is hydrogen, nitro, cyano, halogen, trifluoromethoxy, difluoromethoxy, sulfonamide group, methoxycarbonyl, methoxy, hydroxyl, methyl, amino, Boc protected amino, trifluoromethyl, phenyl;

[0036] Z is halogen-substituted benzene ring, fluorinated alkoxy-substituted benzene ring, methoxy-substituted benzene ring, pyridine ring, pyrimidine ring, cyclohexane, cyclopropane, tetrahydropyran, 1-tert-butoxycarbonylpiperidine, morpholine ring, piperidine ring, tetrahydropyrrole, adamantane;

[0037] substituted or unsubstituted C3 ~ C8 cycloalkyl is selected from: cyclohexane;

[0038] 6-10 membered aromatic heterocycle containing 1-2 N atoms is selected from: pyrimidine ring;

[0039] The substituted or unsubstituted benzo 5-6 membered heterocycle is selected from: naphthalene ring, quinoline ring, quinoxaline, 1,4-benzodioxane, indole ring, benzofuran ring, benzoxazole, bromo-substituted piperonyl ring.

[0040] Further,

[0041] R 1 selected from .

[0042] Specifically, the 2-cyano-2-(9 H -xanthen-9-ylidene)acetamide compound is selected from any one of the following compounds:

[0043] The present application designs a series of novel 2-cyano-2-(9 H -xanthen-9-ylidene)acetamide compounds, which can efficiently bind to ZNF207 and significantly inhibit its activity, and exhibit excellent therapeutic effect in anti-tumor experiments.

[0044] Preferably, the pharmaceutically acceptable salt is a salt of the compound with any one of the following acids or bases: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, carbonic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, malic acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid, mandelic acid or ferulic acid.

[0045] The 2-cyano-2-(9 H -xanthen-9-ylidene)acetamide compound of the present application has the following preparation method:

[0046] (1) Compound a is subjected to Knoevenagel condensation reaction with ethyl cyanoacetate to obtain compound b;

[0047] (2) Compound b is subjected to lithium hydroxide hydrolysis to obtain compound c;

[0048] (3) Compound c is subjected to acid amine condensation reaction with (L) to obtain compound d;

[0049] (4) Compound d is subjected to Boc protection group removal to obtain compound e;

[0050] (5) Compound e is subjected to condensation reaction with or The reaction yields the target compound; wherein compound e reacts with... Nucleophilic substitution reaction occurs; compound e reacts with... An acid-amine condensation reaction occurs; compound e reacts with... A urea-forming reaction occurs; compound e reacts with... A reductive ammoniation reaction occurs.

[0051] Where A, X, n, R 1 The definition is as described above.

[0052] Specifically, compound b is prepared by reacting compound a with ethyl cyanoacetate via a Knauvengel reaction. The catalyst used is selected from piperidine, pyridine, diethylamine, triethylamine, ammonium acetate, or Lewis acid, preferably triethylamine / titanium tetrachloride (TEA / TiCl4). The solvent used is selected from dichloromethane (DCM), tetrahydrofuran (THF), benzene, toluene, 1,4-dioxane, etc. N , N The solvent is dimethylformamide (DMF) or a mixture of both, preferably dichloromethane; the reaction temperature is selected from 25 to 120°C, preferably 70°C.

[0053] Compound c is prepared by ester hydrolysis of compound b. The strong base used is selected from lithium hydroxide (LiOH), sodium hydroxide (NaOH), potassium hydroxide (KOH), with lithium hydroxide being preferred. The solvent used is selected from methanol (MeOH), ethanol (EtOH), water (H2O), or a mixture of any two solvents, with a mixture of ethanol and water being preferred. The reaction temperature is selected from 25 to 85°C, with 25°C being preferred.

[0054] Compound d was prepared from compound c via an acid-amine condensation reaction, using a condensing agent selected from... N , N' -Dicyclohexylcarbodiimide (DCC), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) N , N' -Diisopropylcarbodiimide (DIC), O-(7-azabenzotriazol-1-yl)- hexafluorophosphate N , N , N' , N' Tetramethylureonium (HATU), benzotriazol-1-yl-oxy-tripyrrolylphosphonium hexafluorophosphate (HBTU), benzotriazol-1-yl-oxy-tripyrrolylphosphonium hexafluorophosphate (PyBOP), propylphosphonic anhydride (T3P), preferably HATU; the base used is selected from triethylamine (TEA). N , N- diisopropylethylamine (DIPEA), 4-dimethylaminopyridine, N - methylmorpholine, pyridine, potassium carbonate, sodium carbonate, sodium hydroxide, potassium hydroxide, preferably DIPEA; the solvent used is selected from N , N - dimethylformamide, dimethylsulfoxide (DMSO), acetonitrile (ACN), dichloromethane, chloroform (CHCl3), tetrahydrofuran, 1,4-dioxane, toluene, preferably N , N - dimethylformamide; the reaction temperature is selected from 25-120°C, preferably 25°C.

[0055] After removing the Boc protecting group from compound d, compound e is obtained. The reagent used for removing the protecting group is selected from trifluoroacetic acid (TFA), hydrochloric acid (HCl), trimethylsilyl (TMSI), preferably trifluoroacetic acid; the solvent used is selected from dichloromethane (DCM), ethyl acetate (EA), dioxane, acetonitrile (ACN), preferably dichloromethane.

[0056] Compound e is reacted with or by nucleophilic substitution or acid amine condensation or urea formation reaction or reductive amination reaction to obtain the target compound. For the acid amine condensation reaction: the condensing agent is preferably HATU, the base is preferably DIPEA, the solvent is preferably DMF, and the temperature is preferably 25°C. For the affinity substitution reaction, the base is preferably potassium carbonate, the solvent is preferably DMF, and the temperature is preferably 90°C. For the reductive amination reaction, the reducing agent used is selected from sodium borohydride (NaBH4), sodium triacetoxyborohydride (NaBH(OAc)3), sodium cyanoborohydride (NaBH3CN), preferably sodium cyanoborohydride; the solvent used is selected from methanol, ethanol, dioxane, tetrahydrofuran or a mixed solvent composed of any two thereof, preferably methanol. The reaction temperature is selected from 25-120°C, preferably 80°C.

[0057] The pharmaceutical composition described in the present application comprises the 2-cyano-2-(9 H- xanthen-9-ylidene)acetamide compound and a pharmaceutically acceptable carrier. Common pharmaceutical excipients such as flavors, sweeteners, liquid / solid fillers, diluents, etc. can be added to make common pharmaceutical preparations such as tablets, capsules, syrups, suspensions, injections, etc.

[0058] The 2-cyano-2-(9 H xanthen-9-ylidene)acetamide compound and the pharmaceutical composition thereof described in the present application are used for preparing a ZNF207 inhibitor drug, in particular as an antitumor drug, especially as a drug for treating breast cancer, colorectal cancer, hepatocellular carcinoma, brain glioma, etc.

[0059] The application provides application of the compound or the pharmaceutically acceptable salt thereof or the pharmaceutical composition in preparation of a ZNF207 inhibitor drug.

[0060] The application provides application of the compound or the pharmaceutically acceptable salt thereof in preparation of an anticancer drug, and the cancer is breast cancer, colorectal cancer, hepatocellular carcinoma or brain glioma.

[0061] Beneficial effects: compared with the prior art, the application has the following remarkable advantages:

[0062] The 2-cyano-2-(9 H -xanthen-9-yl)acetamides exhibit nanomolar (nM) binding affinities (K D ) to ZNF207 and show significant anti-tumor activity in cell models. The mechanism of action includes direct inhibition of tumor cell proliferation and effective elimination of tumor stem cells (CSCs) by targeting ZNF207, thereby overcoming tumor drug resistance and inhibiting disease progression. In addition, the synthesis route of the compound is simple and easy to scale up, and has potential clinical transformation value. DETAILED DESCRIPTION

[0063] The technical solutions of the application are further described below in combination with examples.

[0064] Example 1

[0065] N -(2-(1-benzylpiperidin-4-yl)ethyl)-2-cyano-2-(9 H -xanthen-9-yl)acetamide

[0066]

[0067] (1) Preparation of ethyl 2-cyano-2-(9 H -xanthen-9-yl)acetate (b)

[0068] Xanthone (10.19 mmol) was dissolved in dichloromethane (100 mL), TiCl4 (12.7 mmol) was slowly added dropwise, stirred at room temperature for 30 min, then 2-cyanoethyl acetate (20.38 mmol) and triethylamine (132 mmol) were added in sequence, heated to 70℃, and reacted for 8 h. After the reaction was completed by TLC monitoring, water (30 mL) was added, filtered, the filtrate was extracted with dichloromethane (50 mL) for 3 times, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography to obtain 2 g of ethyl 2-cyano-2-(9 H -xanthen-9-yl)acetate in the form of a yellow solid (yield 67.5%).

[0069] (2) Preparation of 2-cyano-2-(9 H - xanthen-9-yl)acetic acid (c)

[0070] Ethyl 2-cyano-2-(9 H - xanthen-9-yl)acetate (7.6 mmol) was dissolved in ethanol (100 mL), and a solution of LiOH H2O (30.3 mmol) in water (10 mL) was added. The reaction was allowed to proceed at room temperature for 4 h, and the reaction was monitored by TLC. After the reaction was completed, the solvent was evaporated under reduced pressure, water (60 mL) was added, and the pH was adjusted to 3-4 using hydrochloric acid. A solid was precipitated, which was filtered to obtain 2-cyano-2-(9 H - xanthen-9-yl)acetic acid 1.8 g as a yellow solid (yield 89.4%).

[0071] (3) Preparation of tert-butyl 4-(2-(2-cyano-2-(9 H - xanthen-9-yl)acetamido)ethyl)piperidine-1-carboxylate (d1)

[0072] 2-cyano-2-(9 H - xanthen-9-yl)acetic acid (4.7 mmol) was dissolved in DMF (N,N-dimethylformamide) (50 mL), and DIPEA (11.7 mmol), HATU (5.1 mmol), and tert-butyl 4-(2-aminoethyl)piperidine-1-carboxylate (4.7 mmol) were sequentially added. The reaction was allowed to proceed at room temperature for 8 h, and the reaction was monitored by TLC. After the reaction was completed, water (50 mL) was added, and extraction was performed three times with EA (100 mL). The organic layer was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography to obtain tert-butyl 4-(2-(2-cyano-2-(9 H - xanthen-9-yl)acetamido)ethyl)piperidine-1-carboxylate 1.7 g as a yellow solid (yield 76.5%).

[0073] (4) Preparation of 2-cyano- N - (2-(piperidin-4-yl)ethyl)-2-(9 H - xanthen-9-yl)acetamide (e1)

[0074] Tert-butyl 4-(2-(2-cyano-2-(9 H - xanthen-9-yl)acetamido)ethyl)piperidine-1-carboxylate (1.05 mmol) was dissolved in DCM (30 mL), and TFA (21 mmol) was added. The reaction was allowed to proceed at room temperature for 2 h, and the reaction was monitored by TLC. After the reaction was completed, the solvent was evaporated under reduced pressure, and a saturated aqueous sodium bicarbonate solution was added. A solid was precipitated, which was filtered to obtain 2-cyano- N - (2-(piperidin-4-yl)ethyl)-2-(9 HXanthen-9-yl)acetamide 353 mg as a white solid (yield 90.1%).

[0075] (5) N -(2-(1 -benzylpiperidin-4-yl)ethyl)-2-cyano-2-(9 H Preparation of (2-(piperidin-4-yl)ethyl)-2-cyano-2-(9

[0076] 2-cyano-2-(9 N -(2-(piperidin-4-yl)ethyl)-2-cyano-2-(9 H Xanthen-9-yl)acetamide was dissolved in DMF (30 mL), K2CO3 (3.16 mmol), benzyl bromide (1.16 mmol) were added successively, heated to 90 °C, and the reaction was allowed to proceed for 8 h. After the reaction was completed as monitored by TLC, water (30 mL) was added, and EA (60 mL) was extracted three times, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography (PE:EA = 1:1) to obtain N -(2-(1 -benzylpiperidin-4-yl)ethyl)-2-cyano-2-(9 H Xanthen-9-yl)acetamide 353 mg as a white solid (yield 90.1%). 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.68 (t, J = 5.5 Hz, 1H), 8.28 (dd, J = 8.1, 1.5 Hz, 1H), 7.72-7.60 (m, 2H), 7.59-7.54 (m, 1H), 7.54-7.48 (m, 1H), 7.48-7.38 (m, 2H), 7.35-7.20 (m, 6H), 3.41 (s, 2H), 3.11 (q, J = 6.6Hz, 2H), 2.73 (d, J = 10.4 Hz, 2H), 1.91-1.70 (m, 2H), 1.62-1.47 (m, 2H), 1.34-1.19 (m, 2H), 1.15- 0.97 (m, 3H). HRMS (ESI) m / z: [M + H] + calcd for C 30 H 30 N3O2, 464.2338; found, 464.2341.

[0077] Example 2

[0078] N -(2-(1-benzoylpiperidin-4-yl)ethyl)-2-cyano-2-(9 H -Xanthan-9-ylidene)acetamide

[0079]

[0080] The preparation method is the same as in Example 1 (1), (2), (3), and (4), except for step (5): the obtained 2-cyano- N -(2-(piperidin-4-yl)ethyl)-2-(9 H 0.8 mmol of 2-xanthan-9-yl)acetamide was dissolved in DMF (30 mL), followed by the addition of DIPEA (2.4 mmol), HATU (0.88 mmol), and benzoic acid (0.8 mmol). The reaction was carried out at room temperature for 8 h. After the reaction was completed by TLC monitoring, 30 mL of water was added, and the mixture was extracted three times with EA (60 mL). The extract was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography (PE:EA = 5:1) to obtain the final product. N -(2-(1-benzylpiperidin-4-yl)ethyl)-2-cyano-2-(9 H 300 mg of 3-xanthon-9-ylidene acetamide was obtained as a white solid (yield 78.5%). 1 H NMR (300 MHz, Chloroform- d ) δ 8.41 (dd, J =8.1, 1.5 Hz, 1H), 7.74 (dd, J = 8.1, 1.5 Hz, 1H), 7.60-7.48 (m, 2H), 7.45-7.30(m, 8H), 7.22-7.11 (m, 1H), 6.37 (t, J = 5.8 Hz, 1H), 4.73-4.49 (m, 1H), 3.82-3.56 (m, 1H), 3.31 (q, J = 6.6 Hz, 2H), 2.99-2.84 (m, 1H), 2.84-2.56 (m, 1H), 1.80-1.54 (m, 2H), 1.55-1.37 (m, 3H), 1.22-1.03 (m, 2H). HRMS (ESI) m / z: [M +Na] + calcd for C 30 H 27N3O3Na, 500.1950; found, 500.1951.

[0081] Example 3

[0082] 2-cyano- N -(2-(1-(quinoline-6-carbonyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0083]

[0084] The procedure of Reference Example 2 was followed with the exception that benzoic acid was replaced by quinoline-6-carboxylic acid to give 2-cyano- N -(2-(1-(quinoline-6-carbonyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (70.7% yield). 1 H NMR (300 MHz, Chloroform- d ) δ 8.93 (dd, J = 4.3, 1.7 Hz, 1H), 8.39 (dd, J = 8.1, 1.5 Hz, 1H), 8.18 (dd, J = 8.3, 1.7 Hz, 1H), 8.09 (d, J = 8.6 Hz, 1H), 7.85 (d, J = 1.8 Hz, 1H), 7.74 (dd, J = 8.1, 1.5 Hz, 1H), 7.65 (dd, J = 8.7, 1.9 Hz, 1H), 7.58-7.41 (m, 3H), 7.36-7.28 (m, 3H), 7.22-7.13 (m, 1H), 6.57 (t, J = 5.8 Hz, 1H), 4.82-4.49 (m, 1H), 3.86-3.51 (m, 1H), 3.32 (q, J= 6.6Hz, 2H), 3.04-2.86 (m, 1H), 2.81-2.60 (m, 1H), 1.82-1.69 (m, 1H), 1.67-1.56(m, 1H), 1.53-1.42 (m, 3H), 1.24-1.02 (m, 2H). HRMS (ESI) m / z: [M + Na] + calcdfor C 33 H 28 N4O3Na, 551.2059; found, 551.2061.

[0085] Example 4

[0086] N -(2-(1-(1 H -Indole-5-carbonyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -xanthen-9-ylidene)acetamide

[0087]

[0088] The preparation method refers to Example 2, except that benzoic acid is replaced by 1 H -indole-5-carboxylic acid to produce N -(2-(1-(1 H -Indole-5-carbonyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (yield 81.3%). 1 H NMR (300 MHz, DMSO- d 6 ) δ 11.27 (s, 1H), 8.78-8.63 (m,1H), 8.29 (d, J = 8.0 Hz, 1H), 7.70-7.62 (m, 2H), 7.61-7.55 (m, 2H), 7.54-7.48(m, 1H), 7.47-7.38 (m, 4H), 7.37-7.22 (m, 1H), 7.10 (d, J= 8.4 Hz, 1H), 6.49(s, 1H), 3.34 (s, 2H), 3.25-3.04 (m, 2H), 2.90-2.59 (m, 2H), 1.77-1.52 (m,2H), 1.39-1.26 (m, 3H), 1.17-0.92 (m, 2H). HRMS (ESI) m / z: [M + Na] + calcd forC 32 H 28 N4O3Na, 539.2059; found, 539.2062。

[0089] Example 5

[0090] 2-cyano- N -(2-(1-phenylsulfonylpiperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0091]

[0092] The preparation method refers to Examples 1, (1), (2), (3), (4) in the difference that step (5): the obtained 2-cyano- N -(2-(piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide (0.8 mmol) was dissolved in DCM (30 mL), DIPEA (2.4 mmol), phenylsulfonyl chloride (0.8 mmol) was added in turn, and the reaction was carried out at room temperature for 8 h. After the reaction was completed by TLC monitoring, water (30 mL) was added, and DCM (60 mL) was extracted 3 times, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography (PE:EA = 5:1) to obtain 2-cyano- N -(2-(1-phenylsulfonylpiperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide 350 mg, white solid (yield 85.3%). 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.60 (t, J = 5.2 Hz, 1H), 8.33-8.21 (m, 1H), 7.85-7.55 (m, 7H), 7.54-7.43 (m, 1H), 7.43 (d, J = 7.5 Hz, 2H),7.31 (d, J= 8.2 Hz, 1H), 7.17 (t, J = 7.7 Hz, 1H), 3.57 (d, J = 11.3 Hz, 2H), 3.07 (q, J = 6.4 Hz, 2H), 1.98 (t, J = 11.6 Hz, 2H), 1.70-1.52 (m, 2H), 1.23 (q, J = 7.0 Hz, 2H), 1.15-0.95 (m, 2H), 0.92-0.76 (m, 1H). HRMS (ESI) m / z: [M + Na] + calcd for C 29 H 27 N3O4NaS, 536.1620; found, 536.1624.

[0093] Example 6

[0094] 2-Cyano- N -(2-(1-(2,3-dihydrobenzo[ b [1,4]dioxane-6-sulfonyl)piperidin-4-yl)ethyl)-2-(9 H -Xanthan-9-ylidene)acetamide

[0095]

[0096] The preparation method is the same as in Example 5, except that benzenesulfonyl chloride is replaced with 2,3-dihydrobenzo[ b [1,4]dioxane-6-sulfonyl chloride, to prepare 2-cyano- N -(2-(1-(2,3-dihydrobenzo[ b [1,4]dioxane-6-sulfonyl)piperidin-4-yl)ethyl)-2-(9 H -Xanton-9-ylidene)acetamide, a white solid (yield 85.7%). 1 HNMR (300 MHz, DMSO- d 6 ) δ 8.60 (t, J = 5.6 Hz, 1H), 8.27 (d, J = 7.8 Hz, 1H), 7.73-7.57 (m, 2H), 7.55-7.44 (m, 2H), 7.41 (d, J= 7.6 Hz, 1H), 7.35 (d, J = 8.3 Hz,1H), 7.26-7.06 (m, 4H), 4.50-4.25 (m, 4H), 3.53 (d, J = 11.2 Hz, 2H), 3.09 (q, J = 6.4 Hz, 2H), 2.11-1.89 (m, 2H), 1.61 (d, J = 12.7 Hz, 2H), 1.30-1.20 (m, 2H),1.14-0.98 (m, 2H), 0.92-0.81 (m, 1H). HRMS (ESI) m / z: [M + Na] + calcd forC 31 H 29 N3O6NaS, 594.1675; found, 594.1665.

[0097] Example 7

[0098] 4-(2-(2-cyano-2-(9) H -Xanton-9-ylidene)acetamido)ethyl)- N -Phenylidene-1-carboxamide

[0099]

[0100] The preparation method is the same as in Example 1 (1), (2), (3), and (4), except for step (5): the obtained 2-cyano- N -(2-(piperidin-4-yl)ethyl)-2-(9 H 4-(2-(2-cyano-2-(9-yl)acetamide (0.7 mmol) was dissolved in DCM (40 mL). DIPEA (1.4 mmol) and triphosgene (0.2 mmol) were added sequentially at 0 °C. The reaction was allowed to proceed for 0.5 h. Then, a dichloromethane solution (3 mL) of aniline (0.7 mmol) was slowly added dropwise. The mixture was brought to room temperature and the reaction continued for another 8 h. After the reaction was completed by TLC monitoring, water (30 mL) was added, and the mixture was extracted three times with DCM (60 mL). The extract was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography (PE:EA = 2:1) to obtain 4-(2-(2-cyano-2-(9-yl)acetamide. H -Xanton-9-ylidene)acetamido)ethyl)- N 230 mg of 1-phenylpiperidine-1-carboxamide was obtained as a white solid (yield 66.7%). 1 H NMR (300 MHz, DMSO- d6 ) δ 8.71 (t, J = 5.5 Hz, 1H), 8.43 (s, 1H),8.36-8.24 (m, 1H), 7.74-7.63 (m, 2H), 7.63-7.56 (m, 1H), 7.55-7.39 (m, 5H),7.30 (t, J = 7.6 Hz, 1H), 7.21 (t, J = 7.8 Hz, 2H), 6.91 (t, J = 7.3 Hz, 1H), 4.08(d, J = 13.1 Hz, 2H), 3.16 (q, J = 6.3 Hz, 2H), 2.63 (t, J = 12.5 Hz, 2H), 1.61 (d, J = 12.6 Hz, 2H), 1.41-1.16 (m, 3H), 1.10-0.83 (m, 2H). HRMS (ESI) m / z: [M +Na] + calcd for C 30 H 28 N4O3Na, 515.2059; found,515.2062.

[0101] Example 8

[0102] 2-cyano- N -(2-(l-(4-nitrobenzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0103]

[0104] Preparation method according to Example 1, except that benzyl bromide is replaced by p-nitrobenzyl bromide, to give 2-cyano- N -(2-(l-(4-nitrobenzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (yield 88.4%). 1 H NMR (400 MHz, DMSO- d 6 ) δ 8.67 (t, J = 5.5 Hz, 1H), 8.33-8.25 (m,1H), 8.19 (d,J = 8.4 Hz, 2H), 7.71-7.63 (m, 2H), 7.62-7.54 (m, 3H), 7.51 (d, J = 8.2 Hz, 1H), 7.48-7.40 (m, 2H), 7.26 (t, J = 7.6 Hz, 1H), 3.55 (s, 2H), 3.12(q, J = 6.6 Hz, 2H), 2.73 (d, J = 10.8 Hz, 2H), 1.86 (t, J = 10.6 Hz, 2H), 1.56(d, J = 9.4 Hz, 2H), 1.29 (q, J = 6.5 Hz, 2H), 1.17-0.99 (m, 3H). HRMS (ESI) m / z:[M + H] + calcd for C 30 H 29 N4O4, 509.2189; found,509.2193.

[0105] Example 9

[0106] 2-cyano- N -(2-(1-(3-nitrobenzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0107]

[0108] The preparation method refers to Example 1, except that benzyl bromide is replaced by 1-(bromomethyl)-3-nitrobenzene to obtain 2-cyano- N -(2-(1-(3-nitrobenzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (yield 89.1%). 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.68 (t, J = 5.4 Hz, 1H), 8.29 (d, J =8.0 Hz, 1H), 8.12 (d, J = 8.3 Hz, 2H), 7.75 (d, J= 7.6 Hz, 1H), 7.63 (dq, J =15.9, 7.8 Hz, 4H), 7.51 (d, J = 8.2 Hz, 1H), 7.45 (d, J = 8.5 Hz, 2H), 7.27 (t, J =7.6 Hz, 1H), 3.56 (s, 2H), 3.13 (q, J = 6.4 Hz, 2H), 2.75 (d, J = 10.7 Hz, 2H),1.99-1.76 (m, 2H), 1.66-1.46 (m, 2H), 1.36-1.21 (m, 2H), 1.19-0.99 (m, 3H).HRMS (ESI) m / z: [M + H] + calcd for C 30 H 29 N4O4, 509.2189; found,509.2190。

[0109] Example 10

[0110] 2-cyano- N -(2-(1-(2-nitrobenzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0111]

[0112] The preparation method refers to Example 1, except that benzyl bromide is replaced by 1-(bromomethyl)-2-nitrobenzene to produce 2-cyano- N -(2-(1-(2-nitrobenzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (yield 83.1%). 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.68 (d, J = 5.5 Hz, 1H), 8.28 (d, J =8.0 Hz, 1H), 7.84 (d, J = 8.1 Hz, 1H), 7.72-7.54 (m, 5H), 7.55-7.37 (m, 4H),7.26 (t, J= 7.6 Hz, 1H), 3.65 (s, 2H), 3.10 (q, 2H), 2.60 (d, J = 10.9 Hz, 2H), 1.83 (t, J = 11.2 Hz, 2H), 1.61-1.42 (m, 2H), 1.34-1.16 (m, 2H), 1.12-0.81 (m,3H). HRMS (ESI) m / z: [M + H] + calcd for C 30 H 29 N4O4, 509.2189; found, 509.2192.

[0113] Example 11

[0114] 2-Cyano-N-(2-(1-(3-cyanobenzyl)piperidin-4-yl)ethyl)-2-(9 H -Xanthan-9-ylidene)acetamide

[0115]

[0116] The preparation method is the same as in Example 1, except that benzyl bromide is replaced with 3-(bromomethyl)benzonitrile to obtain 2-cyano- N -(2-(1-(3-cyanobenzyl)piperidin-4-yl)ethyl)-2-(9 H -Xanton-9-ylidene)acetamide, a white solid (yield 91.2%). 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.68 (t, J = 5.5 Hz, 1H), 8.28 (d, J =7.9 Hz, 1H), 7.80-7.69 (m, 2H), 7.68-7.61 (m, 3H), 7.58 (d, J = 8.1 Hz, 1H),7.55-7.49 (m, 2H), 7.48-7.37 (m, 2H), 7.26 (t, J = 7.6 Hz, 1H), 3.48 (s, 2H), 3.12 (q, J = 6.5 Hz, 2H), 2.71 (d, J= 10.8 Hz, 2H), 1.94-1.72 (m, 2H), 1.67-1.44 (m, 2H), 1.37-1.19 (m, 2H), 1.17-0.95 (m, 3H). HRMS (ESI) m / z: [M + Na] + calcd for C 31 H 28 N4O2Na, 511.2110; found,511.2112.

[0117] Example 12

[0118] 2-cyano- N -(2-(1-(4-fluorobenzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0119]

[0120] Preparation method according to Example 1, except that benzylbromide is replaced by 1-(bromomethyl)-4-fluorobenzene, to give 2-cyano- N -(2-(1-(4-fluorobenzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (yield 83.4%). 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.68 (t, J = 5.5 Hz, 1H), 8.35-8.21 (m,1H), 7.73-7.58 (m, 2H), 7.62-7.53 (m, 1H), 7.51 (d, J = 8.2 Hz, 1H), 7.48-7.37(m, 2H), 7.36-7.20 (m, 3H), 7.13 (t, J = 8.8 Hz, 2H), 3.39 (s, 2H), 3.12 (q, J =6.5 Hz, 2H), 2.72 (d, J = 10.8 Hz, 2H), 1.79 (t, J = 10.5 Hz, 2H), 1.62-1.44 (m,2H), 1.35-1.22 (m, 2H), 1.14-0.99 (m, 3H). HRMS (ESI) m / z: [M + H]+ C 30 H 29 N3O2F, 482.2244; found,482.2248.

[0121] Example 13

[0122] 2-cyano- N -(2-(1-(3-fluorobenzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0123]

[0124] The procedure of Reference Example 1 was followed except that 1-(bromomethyl)-3- fluorobenzene was used in place of benzyl bromide to give 2-cyano- N -(2-(1-(3-fluorobenzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (82.9% yield). 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.68 (t, J = 5.5 Hz, 1H), 8.29 (dd, J =8.1, 1.5 Hz, 1H), 7.66 (tdd, J = 8.1, 6.9, 1.6 Hz, 3H), 7.63-7.53 (m, 1H), 7.51(dd, J = 8.3, 1.2 Hz, 1H), 7.45 (ddd, J = 8.1, 2.7, 1.1 Hz, 2H), 7.37 (ddd, J = 14.1, 5.4, 3.6 Hz, 2H), 7.32-7.20 (m, 1H), 7.19-7.00 (m, 3H), 3.45 (s, 2H),3.12 (q, J = 6.6 Hz, 2H), 2.74 (d, J = 10.5 Hz, 2H), 1.83 (s, 2H), 1.55 (d, J = 9.2 Hz, 2H), 1.28 (q, J= 6.6 Hz, 2H), 1.18-0.94 (m, 3H). HRMS (ESI) m / z: [M +H] + calcd for C 30 H 29 N3O2F, 482.2244; found, 482.2247.

[0125] Example 14

[0126] N -(2-(1-(4-chlorobenzyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -Xanthan-9-ylidene)acetamide

[0127]

[0128] The preparation method is the same as in Example 1, except that benzyl bromide is replaced with 1-(bromomethyl)-4-chlorobenzene to obtain... N -(2-(1-(4-chlorobenzyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -Xanton-9-ylidene)acetamide, a white solid (yield 82.4%). 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.68 (t, J = 5.5 Hz, 1H), 8.35-8.21 (m,1H), 7.73-7.58 (m, 2H), 7.62-7.53 (m, 1H), 7.51 (d, J = 8.2 Hz, 1H), 7.48-7.37(m, 2H), 7.36-7.20 (m, 3H), 7.13 (t, J = 8.8 Hz, 2H), 3.39 (s, 2H), 3.12 (q, J =6.5 Hz, 2H), 2.72 (d, J = 10.8 Hz, 2H), 1.79 (t, J = 10.5 Hz, 2H), 1.62-1.44 (m,2H), 1.35-1.22 (m, 2H), 1.14-0.99 (m, 3H). HRMS (ESI) m / z: [M + H] + calcd forC 30 H 29N3O2Cl, 498.1948; found,498.1950.

[0129] Example 15

[0130] N -(2-(l-(3-chlorobenzyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -xanthen-9-ylidene)acetamide

[0131]

[0132] The procedure of Reference Example 1 was followed except that 1-(bromomethyl)-3- chlorobenzene was used instead of benzyl bromide to give N -(2-(l-(3-chlorobenzyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (yield 82.7%). 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.56 (t, J = 5.5 Hz, 1H), 8.23-8.13 (m, 1H), 7.64-7.53 (m, 2H), 7.52-7.45 (m, 1H), 7.41 (d, J = 8.2 Hz, 1H), 7.38-7.27 (m, 2H), 7.27-7.20 (m, 3H), 7.20-7.06 (m, 2H), 3.34 (s, 2H), 3.02 (q, J =6.6 Hz, 2H), 2.63 (d, J = 10.7 Hz, 2H), 1.84-1.57 (m, 2H), 1.56-1.35 (m, 2H),1.30-1.10 (m, 2H), 1.07-0.85 (m, 3H). HRMS (ESI) m / z: [M + H] + calcd forC 30 H 29 N3O2Cl, 498.1948; found,498.1951.

[0133] Example 16

[0134] N -(2-(l-(3-chlorobenzyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -xanthen-9-ylidene)acetamide

[0135]

[0136] Preparation method according to example 1, except that benzyl bromide was replaced by 1-(bromomethyl)-2-chlorobenzene to give N -(2-(1-(2-chlorobenzyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (yield 82.8%). 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.69 (t, J = 5.4 Hz, 1H), 8.29 (d, J =8.0 Hz, 1H), 7.75-7.55 (m, 3H), 7.55-7.37 (m, 5H), 7.37-7.20 (m, 3H), 3.50(s, 2H), 3.12 (d, J = 6.4 Hz, 2H), 2.76 (d, J = 10.7 Hz, 2H), 1.92 (t, J = 10.9 Hz,2H), 1.71-1.45 (m, 2H), 1.29 (q, J = 6.4 Hz, 2H), 1.20-0.98 (m, 3H). HRMS(ESI) m / z: [M + H] + calcd for C 30 H 29 N3O2Cl, 498.1948; found,498.1950.

[0137] Example 17

[0138] N -(2-(1-(4-bromobenzyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -xanthen-9-ylidene)acetamide

[0139]

[0140] Preparation method according to example 1, except that benzyl bromide was replaced by 1-(bromomethyl)-4-bromobenzene to give N -(2-(1-(4-bromobenzyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 HXanthen-9-yl)acetamide as a white solid (yield 79.8%). 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.67 (t, J = 5.5 Hz, 1H), 8.28 (d, J =8.0 Hz, 1H), 7.65 (q, J = 8.0 Hz, 2H), 7.57 (d, J = 7.6 Hz, 1H), 7.54-7.36 (m,5H), 7.32-7.13 (m, 3H), 3.38 (s, 2H), 3.12 (q, J = 6.5 Hz, 2H), 2.71 (d, J = 10.8Hz, 2H), 1.80 (t, J = 10.7 Hz, 2H), 1.64-1.44 (m, 2H), 1.27 (q, J = 6.6 Hz, 2H),1.15-0.91 (m, 3H). HRMS (ESI) m / z: [M + H] + calcd for C 30 H 29 N3O2Br, 542.1443;found,542.1446.

[0141] Example 18

[0142] 2-cyano- N -(2-(1-(4-(difluoromethoxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-yl)acetamide

[0143]

[0144] The preparation method refers to Example 1, except that benzyl bromide is replaced by 1-(bromomethyl)-4-(difluoromethoxy)benzene to obtain 2-cyano- N -(2-(1-(4-(difluoromethoxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-yl)acetamide as a white solid (yield 75.4%). 1 H NMR (300 MHz, DMSO-d6) δ 8.67 (t, J= 5.5Hz, 1H), 8.38-8.20 (m, 1H), 7.78-7.61 (m, 2H), 7.60-7.54 (m, 1H), 7.54-7.48(m, 1H), 7.48-7.39 (m, 2H), 7.38-7.29 (m, 2H), 7.29-7.23 (m, 1H), 7.22-6.92(m, 3H), 3.41 (s, 2H), 3.12 (q, J = 6.5 Hz, 2H), 2.73 (d, J = 10.7 Hz, 2H), 1.79(t, J = 11.5 Hz, 2H), 1.54 (d, J = 8.4 Hz, 2H), 1.28 (q, J = 6.8 Hz, 2H), 1.18-0.92(m, 3H). HRMS (ESI) m / z: [M + H] + calcd for C 31 H 30 N3O3F2, 530.2255; found, 530.2259.

[0145] Example 19

[0146] 2-Cyano- N -(2-(1-(4-(trifluoromethoxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -Xanton-9-ylidene)acetamide

[0147]

[0148] The preparation method is the same as in Example 1, except that benzyl bromide is replaced with 1-(bromomethyl)-4-(trifluoromethoxy)benzene to obtain 2-cyano- N -(2-(1-(4-(trifluoromethoxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H 1,3-(xanton-9-ylidene)acetamide, is a white solid (yield 76.4%). 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.67 (t, J = 5.6 Hz, 1H), 8.29 (dd, J= 8.1, 1.5 Hz, 1H), 7.72-7.60 (m, 2H), 7.60-7.54 (m, 1H), 7.51(d, J = 8.1 Hz, 1H), 7.48-7.36 (m, 4H), 7.35-7.19 (m, 3H), 3.44 (s, 2H), 3.12(q, J = 6.6 Hz, 2H), 2.72 (d, J = 10.8 Hz, 2H), 1.80 (t, J = 10.9 Hz, 2H), 1.54 (d, J = 8.4 Hz, 2H), 1.28 (q, J = 6.5 Hz, 2H), 1.17-0.93 (m, 3H). HRMS (ESI) m / z: [M+ H] + calcd for C 31 H 29 N3O3F3, 548.2161; found,548.2165.

[0149] Example 20

[0150] 2-cyano- N -(2-(1-(4-sulfamoylbenzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0151]

[0152] The preparation method refers to Example 1, except that benzyl bromide is replaced by 4-(bromomethyl)benzenesulfonamide to obtain 2-cyano- N -(2-(1-(4-sulfamoylbenzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (yield 50.1%). 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.67 (t, J = 5.6 Hz, 1H), 8.29 (d, J = 8.0 Hz, 1H), 7.78 (d, J = 7.9 Hz, 2H), 7.65 (q, J= 7.5 Hz, 2H), 7.58 (d, J =7.7 Hz, 1H), 7.54-7.38 (m, 5H), 7.37-7.18 (m, 3H), 3.49 (s, 2H), 3.12 (q, J =6.5 Hz, 2H), 2.73 (d, J = 10.6 Hz, 2H), 1.95-1.72 (m, 2H), 1.55 (d, J = 8.8 Hz,2H), 1.36-1.24 (m, 2H), 1.17-0.98 (m, 3H). HRMS (ESI) m / z: [M + H] + calcd forC 30 H 31 N4O4S, 543.2066; found, 543.2065.

[0153] Example 21

[0154] 3-((4-(2-(2-cyano-2-(9H-xanton-9-ylidene)acetamido)ethyl)piperidin-1-yl)methyl)benzoate

[0155]

[0156] The preparation method is the same as in Example 1, except that benzyl bromide is replaced with methyl 3-(bromomethyl)benzoate to obtain 3-((4-(2-(2-cyano-2-(9 H methyl benzoate (-xanton-9-ylidene)acetamido)ethyl)piperidin-1-yl)methyl)benzoate, as a white solid (yield 54.2%). 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.73 (t, J = 5.6 Hz, 1H),8.34-8.23 (m, 1H), 7.96-7.78 (m, 2H), 7.72-7.61 (m, 2H), 7.60-7.54 (m, 2H),7.54-7.36 (m, 4H), 7.25 (t, J = 7.9 Hz, 1H), 3.86 (s, 3H), 3.48 (s, 2H), 3.11(q, J= 6.6 Hz, 2H), 2.73 (s, 2H), 1.97-1.70 (m, 2H), 1.68-1.46 (m, 2H), 1.40-1.20 (m, 2H), 1.16-0.93 (m, 3H). HRMS (ESI) m / z: [M + H] + calcd for C 32 H 32 N3O4,522.2393; found,522.2396。

[0157] Example 22

[0158] 2-cyano- N -(2-(1-(4-methoxybenzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0159]

[0160] The preparation method refers to Example 1, except that benzyl bromide is replaced by 1-(bromomethyl)-4-methoxybenzene to obtain 2-cyano- N -(2-(1-(4-methoxybenzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (yield 63.1%). 1 H NMR (300 MHz, DMSO-d6) δ 8.68 (t, J = 5.5 Hz, 1H), 8.28(d, J = 8.0 Hz, 1H), 7.65 (q, J = 8.1, 6.8 Hz, 2H), 7.57 (d, J = 7.7 Hz, 1H), 7.51(d, J = 8.2 Hz, 1H), 7.50-7.37 (m, 2H), 7.25 (t, J = 7.6 Hz, 1H), 7.17 (d, J = 8.1Hz, 2H), 6.87 (d, J = 8.1 Hz, 2H), 3.73 (s, 3H), 3.35 (s, 2H), 3.11 (q, J = 6.5Hz, 2H), 2.71 (d, J= 10.7 Hz, 2H), 1.90-1.65 (m, 2H), 1.63-1.42 (m, 2H), 1.36-1.17 (m, 2H), 1.14-0.91 (m, 3H). HRMS (ESI) m / z: [M + H] + calcd for C 31 H 32 N3O3,494.2444; found,494.2448。

[0161] Example 23

[0162] 2-cyano- N -(2-(1-(4-hydroxybenzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0163]

[0164] Preparation method refers to Example 1, (1), (2), (3), (4), the difference is that step (5): the obtained 2-cyano- N -(2-(piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide (0.8 mmol) is dissolved in DMF (30 mL), and then p-hydroxybenzaldehyde (0.8 mmol) and sodium cyanoborohydride (2 mmol) are added in sequence, and the reaction is carried out at 80°C for 8h. After the reaction is completed by TLC monitoring, the solvent is evaporated under reduced pressure, saturated sodium bicarbonate aqueous solution (30 mL) is added, and DCM (60 mL) is extracted for 3 times, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography (DCM:MeOH = 25:1) to obtain 2-cyano- N -(2-(1-(4-hydroxybenzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide 263 mg, white solid (yield 68.6%). 1 HNMR (300 MHz, DMSO- d 6 ) δ 9.73 (s, 1H), 8.71 (t, J = 5.6 Hz, 1H), 8.27 (d, J = 8.0Hz, 1H), 7.74-7.47 (m, 4H), 7.47-7.37 (m, 2H), 7.36-7.07 (m, 3H), 6.79 (d, J= 7.9 Hz, 2H), 4.37-3.62 (m, 1H), 3.57-3.25 (m, 3H), 3.24-2.80 (m, 4H), 1.82-1.52 (m, 2H), 1.50-0.99 (m, 5H). HRMS (ESI) m / z: [M + H] + calcd for C 30 H 30 N3O3,480.2287; found,480.2290。

[0165] Example 24

[0166] 2-cyano- N -(2-(1-(2-hydroxybenzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0167]

[0168] The preparation method refers to Example 23, except that p-hydroxybenzaldehyde is replaced by 2-hydroxybenzaldehyde to prepare 2-cyano- N -(2-(1-(2-hydroxybenzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (yield 63.1%). 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.69 (t, J = 5.6 Hz, 1H), 8.29 (dd, J = 8.1, 1.5 Hz, 1H), 7.73-7.61 (m, 2H), 7.61-7.56 (m, 1H), 7.55-7.38 (m, 3H),7.32-7.22 (m, 1H), 7.15-7.01 (m, 2H), 6.79-6.66 (m, 2H), 3.62 (s, 2H), 3.13(q, J = 6.5 Hz, 2H), 2.93-2.75 (m, 2H), 1.94 (t, J = 10.8 Hz, 2H), 1.61 (d, J = 9.6Hz, 2H), 1.30 (q, J= 6.5 Hz, 2H), 1.18-0.94 (m, 3H). [M + H] + calcd for C 30 H 30 N3O3, 480.2287; found,480.2291.

[0169] Example 25

[0170] N -(2-(1-(3-aminobenzyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -xanthen-9-ylidene)acetamide

[0171]

[0172] The procedure of Reference Example 1 was followed except that benzyl bromide was replaced by 4-aminobenzyl bromide to give N -(2-(1-(3-aminobenzyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (40.2% yield). 1 H NMR (300 MHz, Chloroform- d ) δ 8.53-8.27 (m, 1H), 7.72 (d, J = 8.0Hz, 1H), 7.59-7.42 (m, 2H), 7.33 (q, J = 6.9 Hz, 3H), 7.12 (dt, J = 21.5, 7.7 Hz,2H), 6.67 (d, J = 5.0 Hz, 2H), 6.57 (dd, J = 8.0, 2.4 Hz, 1H), 5.83 (d, J = 5.9Hz, 1H), 3.63 (s, 2H), 3.39 (s, 2H), 3.32 (q, J = 7.0 Hz, 2H), 2.86 (d, J = 11.2Hz, 2H), 1.88 (t, J = 11.1 Hz, 2H), 1.60 (d, J = 11.1 Hz, 2H), 1.40 (q, J= 6.9 Hz,2H), 1.31-1.14 (m, 3H). HRMS (ESI) m / z: [M + H] + calcd for C 30 H 31 N4O2, 479.2447; found, 479.2441.

[0173] Example 26

[0174] tert-butyl(4-((4-(2-(2-cyano-2-(9H-xanton-9-ylidene)acetamido)ethyl)piperidin-1-yl)methyl)phenyl)carbamate

[0175]

[0176] The preparation method is the same as in Example 1, except that benzyl bromide is replaced with tert-butyl(4-(bromomethyl)phenyl)carbamate to obtain tert-butyl(4-((4-(2-(2-cyano-2-(9 H -Xanton-9-ylidene)acetamido)ethyl)piperidin-1-yl)methyl)phenyl)carbamate, as a white solid (yield 55.3%). 1 H NMR (400 MHz, DMSO- d 6 ) δ 9.29 (s, 1H), 8.67 (t, J = 5.6 Hz, 1H), 8.28 (dd, J = 8.1, 1.5 Hz, 1H), 7.70-7.62(m, 2H), 7.62-7.54 (m, 1H), 7.51 (dd, J = 8.4, 1.2 Hz, 1H), 7.47-7.42 (m, 2H), 7.42-7.35 (m, 2H), 7.29-7.21 (m, 1H), 7.13 (d, J = 8.3 Hz, 2H), 3.35 (s, 2H), 3.11 (q, J = 6.6 Hz, 2H), 2.71 (d, J = 10.7 Hz, 2H), 1.82-1.68 (m, 2H), 1.56-1.49(m, 2H), 1.49 (s, 9H), 1.27 (q, J= 6.6 Hz, 2H), 1.11-0.97 (m, 3H). HRMS (ESI)m / z: [M + H] + calcd for C 35 H 39 N4O4, 579.2971; found,579.2974.

[0177] Example 27

[0178] tert-butyl (3-((4-(2-(2-cyano-2-(9H-xanthen-9-ylidene)acetamido)ethyl)piperidin-1- yl)methyl)phenyl)carbamate H - xanthen-9-ylidene)acetamido)ethyl)piperidin-1-yl)methyl)phenyl)carbamate as a white solid (yield 54.3%).

[0179]

[0180] Example 1 with the exception that benzyl bromide was replaced by tert-butyl (3- (bromomethyl)phenyl)carbamate to give tert-butyl (3-((4-(2-(2-cyano-2-(9H-xanthen-9- ylidene)acetamido)ethyl)piperidin-1-yl)methyl)phenyl)carbamate as a white solid (yield 54.3%). 1 H NMR (400 MHz, DMSO- d 6 ) δ 9.31 (s, 1H), 8.68 (t, J = 5.5 Hz, 1H), 8.32-8.24 (m, 1H), 7.72-7.63 (m, 2H),7.62-7.55 (m, 1H), 7.54-7.49 (m, 1H), 7.49-7.40 (m, 3H), 7.34-7.22 (m, 2H),7.17 (t, J = 7.8 Hz, 1H), 6.87 (d, J = 7.5 Hz, 1H), 3.36 (s, 2H), 3.12 (q, J = 6.7Hz, 2H), 2.74 (d, J = 11.1 Hz, 2H), 1.80 (s, 2H), 1.60-1.51 (m, 2H), 1.47 (s,9H), 1.33-1.24 (m, 2H), 1.15-0.96 (m, 3H). HRMS (ESI)m / z: [M + H] + calcd for C 35 H39 N4O4, 579.2971; found,579.2962.

[0181] Example 28

[0182] 2-cyano-N-(2-(l-(3,4-dimethoxybenzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0183]

[0184] The preparation method refers to Example 1, except that benzyl bromide is replaced by 4-(bromomethyl)-l,2-dimethoxybenzene to produce 2-cyano- N -(2-(l-(3,4-dimethoxybenzyl)piperidin-4-yl)ethyl)-2-(9H-xanthen-9-ylidene)acetamide as a white solid (yield 58.3%). 1 H NMR (300 MHz, DMSO-d6) δ 8.69 (t, J = 5.3 Hz,1H), 8.28 (d, J = 8.0 Hz, 1H), 7.60 (dd, J = 24.6, 7.7 Hz, 3H), 7.49 (d, J = 8.3Hz, 1H), 7.43 (d, J = 8.7 Hz, 2H), 7.24 (t, J = 7.5 Hz, 1H), 6.97-6.83 (m, 2H),6.78 (d, J = 8.2 Hz, 1H), 3.73 (s, 6H), 3.39 (s, 2H), 3.11 (q, J = 6.4 Hz, 2H),2.77 (d, J = 10.9 Hz, 2H), 1.84 (s, 2H), 1.63-1.41 (m, 2H), 1.35-1.23 (m, 2H),1.17-0.98 (m, 3H). HRMS (ESI) m / z: [M + H] + calcd for C 32 H 34 N3O4, 524.2549;found,524.2553.

[0185] Example 29

[0186] N -(2-(l-(3,5-bis(trifluoromethyl)benzyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -xanthen-9-ylidene)acetamide

[0187]

[0188] Preparation Method Reference Example 1, except that benzyl bromide was replaced by l-(bromomethyl)-3,5-bis(trifluoromethyl)benzene to produce N -(2-(l-(3,5-bis(trifluoromethyl)benzyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (yield 66.7%). 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.69 (t, J = 5.5 Hz, 1H), 8.29 (d, J = 8.0 Hz, 1H), 7.98 (s, 3H), 7.66 (d, J = 7.9 Hz, 2H), 7.57 (d, J = 7.8 Hz, 1H), 7.51 (d, J = 8.2 Hz, 1H), 7.44 (d, J = 7.9 Hz, 2H), 7.27 (t, J =7.7 Hz, 1H), 3.63 (s, 2H), 3.13 (q, J = 6.5 Hz, 2H), 2.74 (d, J = 10.8 Hz, 2H),1.88 (t, J = 10.5 Hz, 2H), 1.69-1.46 (m, 2H), 1.37-1.21 (m, 2H), 1.19-0.98 (m,3H)。HRMS (ESI) m / z: [M + H] + calcd for C 32 H 28 N3O2F6, 600.2086; found,600.2089.

[0189] Example 30

[0190] N- (2-( 1 -(3-chloro-5-methoxybenzyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H - xanthen-9-ylidene)acetamide

[0191]

[0192] Preparation Method Reference Example 1, except that benzyl bromide was replaced with 1-(bromomethyl)-3-chloro-5-methoxybenzene to produce N - (2-( 1 -(3-chloro-5-methoxybenzyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H - xanthen-9-ylidene)acetamide as a white solid (yield 68.3%). 1 H NMR (300 MHz, DMSO-d6) δ 8.67 (d, J = 5.7 Hz,1H), 8.34-8.22 (m, 1H), 7.73-7.58 (m, 2H), 7.58 (d, J = 8.5 Hz, 1H), 7.51 (d, J =8.2 Hz, 1H), 7.44 (dd, J = 12.1, 5.8 Hz, 2H), 7.26 (t, J = 7.6 Hz, 1H), 6.90 (dd, J = 5.1, 2.7 Hz, 2H), 6.81 (s, 1H), 3.76 (s, 3H), 3.39 (s, 2H), 3.12 (q, J = 6.5Hz, 2H), 2.72 (d, J = 10.6 Hz, 2H), 1.79 (t, J = 10.9 Hz, 2H), 1.65-1.45 (m, 2H),1.36-1.19 (m, 2H), 1.18-0.94 (m, 3H). HRMS (ESI) m / z: [M + H] + calcd forC 31 H 31 N3O3Cl, 528.2054; found,528.2059.

[0193] Example 31

[0194] 2-cyano- N- (2-(1-(3,5-dimethoxybenzyl)piperidin-4-yl)ethyl)-2-(9 H - xanthen-9-ylidene)acetamide

[0195]

[0196] Preparation Method Reference Example 1, except that benzyl bromide was replaced with 1-(bromomethyl)-3,5-dimethoxybenzene to produce 2-cyano- N - (2-(1-(3,5-dimethoxybenzyl)piperidin-4-yl)ethyl)-2-(9 H - xanthen-9-ylidene)acetamide as a white solid (yield 51.6%). 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.68 (t, J = 5.5 Hz,1H), 8.33-8.24 (m, 1H), 7.72-7.58 (m, 2H), 7.58 (d, J = 7.2 Hz, 1H), 7.51 (d, J =8.2 Hz, 1H), 7.49-7.37 (m, 2H), 7.26 (t, J = 7.6 Hz, 1H), 6.44 (d, J = 2.3 Hz,2H), 6.36 (t, J = 2.4 Hz, 1H), 3.72 (s, 6H), 3.34 (s, 2H), 3.12 (q, J = 6.6 Hz,2H), 2.74 (d, J = 10.8 Hz, 2H), 1.79 (t, J = 10.6 Hz, 2H), 1.63-1.46 (m, 2H),1.28 (q, J = 6.4 Hz, 2H), 1.18 - 0.97 (m, 3H). HRMS (ESI) m / z: [M + H] + calcd forC 32 H 34 N3O4, 524.2549; found,524.2554.

[0197] Example 32

[0198] 2-cyano-N - (2-(l-(4-hydroxy-3-methoxybenzyl)piperidin-4-yl)ethyl)-2-(9 H - xanthen-9-ylidene)acetamide

[0199]

[0200] Preparation Method Reference Example 23, except that p-hydroxybenzaldehyde was replaced with 4-hydroxy-3-methoxybenzaldehyde to produce 2-cyano- N - (2-(l-(4-hydroxy-3-methoxybenzyl)piperidin-4-yl)ethyl)-2-(9 H - xanthen-9-ylidene)acetamide as a white solid (yield 55.2%). 1 H NMR (300 MHz, Chloroform- d ) δ 8.41 (dd, J = 8.1, 1.5 Hz, 1H), 7.71 (dd, J = 8.1, 1.5 Hz, 1H), 7.60-7.42 (m, 2H), 7.39-7.26 (m, 4H), 7.21-7.10 (m, 1H), 6.93 (s, 1H), 6.82 (d, J = 8.0 Hz, 1H), 6.74(dd, J = 8.0, 1.8 Hz, 1H), 5.93 (s, 1H), 3.87 (s, 3H), 3.46 (s, 2H), 3.32 (q, J = 6.8 Hz, 2H), 2.91 (d, J = 11.2 Hz, 2H), 1.95 (t, J = 11.3 Hz, 2H), 1.63 (d, J =11.7 Hz, 2H), 1.42 (q, J = 6.9 Hz, 2H), 1.36-1.21 (m, 3H). HRMS (ESI) m / z: [M +H] + calcd for C 31 H 32 N3O4, 510.2393; found,510.2396.

[0201] Example 33

[0202] 2-cyano- N-(2-(l-(pyrimidin-2-ylmethyl)piperidin-4-yl)ethyl)-2-(9 H - xanthen-9-ylidene)acetamide

[0203]

[0204] Preparation Method Reference Example 1, except that benzyl bromide was replaced by 2-(bromomethyl)pyrimidine to produce 2-cyano- N -(2-(l-(pyrimidin-2-ylmethyl)piperidin-4-yl)ethyl)-2-(9 H - xanthen-9-ylidene)acetamide as a white solid (yield 71.3%). 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.77 (d, J = 4.9 Hz, 2H), 8.66 (t, J =5.3 Hz, 1H), 8.28 (d, J = 8.0 Hz, 1H), 7.65 (d, J = 8.0 Hz, 2H), 7.57 (d, J = 7.6Hz, 1H), 7.51 (d, J = 8.2 Hz, 1H), 7.44 (d, J = 8.4 Hz, 2H), 7.39 (t, J = 4.6 Hz,1H), 7.25 (t, J = 7.7 Hz, 1H), 3.65 (s, 2H), 3.11 (q, J = 6.5 Hz, 2H), 2.82 (d, J =11.0 Hz, 2H), 1.98 (t, J = 10.6 Hz, 2H), 1.64-1.42 (m, 2H), 1.27 (q, J = 6.6 Hz,2H), 1.15-0.89 (m, 3H). HRMS (ESI) m / z: [M + H] + calcd for C 28 H 28 N5O2, 466.2243;found,466.2246.

[0205] Example 34

[0206] N -2-(adamantan-l-yl)-N-(2-(l-(2-(adamantan-l-yl)acetyl)piperidin-4-yl)ethyl)-2- cyanopropanamide H -2-(adamantan-l-yl)-N-(2-(l-(2-(adamantan-l-yl)acetyl)piperidin-4-yl)ethyl)-2- cyanopropanamide

[0207]

[0208] Reference to the preparation method of Example 2, except that benzoic acid was replaced by 2-(adamantan-l-yl)acetic acid to give N -2-(adamantan-l-yl)-N-(2-(l-(2-(adamantan-l-yl)acetyl)piperidin-4-yl)ethyl)-2- cyanopropanamide H -2-(adamantan-l-yl)-N-(2-(l-(2-(adamantan-l-yl)acetyl)piperidin-4-yl)ethyl)-2- cyanopropanamide as a white solid (yield 71.4%). 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.69 (t, J = 5.5 Hz, 1H), 8.29 (d, J = 8.0 Hz, 1H), 7.66 (q, J = 7.9 Hz, 2H), 7.59 (d, J = 7.7 Hz, 1H), 7.56-7.38 (m, 3H), 7.28 (t, J = 7.6 Hz, 1H), 4.41 (d, J = 12.9 Hz, 1H), 3.92 (d, J = 13.5 Hz, 1H), 3.13 (q, J = 6.1 Hz, 2H), 2.83 (t, J = 12.5 Hz, 1H), 2.41-2.25 (m, 1H), 2.07 (q, J = 13.5 Hz, 2H), 1.90 (s, 3H), 1.68-1.54 (m, 14H), 1.35-1.20 (m, 3H), 1.00-0.73 (m, 2H). HRMS (ESI) m / z: [M + Na] + calcd for C 35 H 39 N3O3Na, 572.2889; found, 572.2891.

[0209] Example 35

[0210] 2-cyano-N-(2-(1-(cyclohexylmethyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0211]

[0212] Preparation Method Reference Example 1, except that benzyl bromide was replaced with (bromomethyl)cyclohexane to produce 2-cyano- N -(2-(1-(cyclohexylmethyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (65.3% yield). 1 H NMR (300 MHz, Chloroform- d ) δ 8.41 (d, J = 8.0 Hz, 1H), 7.72 (d, J = 8.0 Hz, 1H), 7.52 (q, J = 8.5 Hz, 2H), 7.40-7.28 (m, 3H), 7.17 (t, J = 7.6Hz, 1H), 5.91 (t, J = 5.4 Hz, 1H), 3.33 (q, J = 6.8 Hz, 2H), 2.91 (d, J = 11.2Hz, 2H), 2.16 (d, J = 6.9 Hz, 2H), 1.88 (t, J = 11.5 Hz, 2H), 1.80-1.70 (m, 3H),1.70-1.57 (m, 4H), 1.42 (q, J = 7.1 Hz, 3H), 1.36-1.23 (m, 3H), 1.22-1.04 (m,3H), 0.87 (q, J = 11.2, 10.7 Hz, 2H). HRMS (ESI) m / z: [M + H] + calcd forC 30 H 36 N3O2, 470.2808; found,470.2810.

[0213] Example 36

[0214] 2-cyano- N-2-(9 H -2-(9

[0215]

[0216] Reference to the preparation method of Example 1, except that benzyl bromide was replaced by 2-(bromomethyl)naphthalene to give 2-cyano- N -2-(9 H -2-(9 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.67 (t, J = 5.5 Hz, 1H), 8.35-8.21 (m,1H), 7.96-7.83 (m, 3H), 7.77 (s, 1H), 7.71-7.62 (m, 2H), 7.62-7.54 (m, 1H),7.54-7.38 (m, 6H), 7.32-7.18 (m, 1H), 3.58 (s, 2H), 3.12 (q, J = 6.6 Hz, 2H),2.79 (d, J = 10.9 Hz, 2H), 1.99-1.74 (m, 2H), 1.56 (d, J = 9.1 Hz, 2H), 1.29 (q, J = 6.6 Hz, 2H), 1.18-0.96 (m, 3H). HRMS (ESI) m / z: [M + H] + calcd for C 34 H 32 N3O2,514.2495; found,514.2499.

[0217] Example 37

[0218] 2-cyano-N-(2-(1-(quinolin-8-ylmethyl)piperidin-4-yl)ethyl)-2-(9 H -2-(9

[0219]

[0220] The preparation method refers to Example 1, except that benzyl bromide is replaced by 8-(bromomethyl)quinoline to prepare 2-cyano-N-(2-(l-(quinolin-8-ylmethyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-yl)acetamide as a white solid (yield 83.2%). 1 H NMR (300 MHz, DMSO- d 6 ) δ 9.06-8.83 (m, 1H), 8.69 (t, J = 5.6Hz, 1H), 8.40 (d, J = 8.3 Hz, 1H), 8.32-8.22 (m, 1H), 8.07-7.75 (m, 2H), 7.73-7.62 (m, 3H), 7.62-7.48 (m, 3H), 7.47-7.38 (m, 2H), 7.31-7.20 (m, 1H), 3.34(s, 2H), 3.13 (q, J = 6.5 Hz, 2H), 3.06-2.76 (m, 2H), 2.33-1.88 (m, 2H), 1.61(d, J = 11.2 Hz, 2H), 1.46-0.96 (m, 5H). HRMS (ESI) m / z: [M + H] + calcd forC 33 H 31 N4O2, 515.2447; found,515.2449.

[0221] Example 38

[0222] 2-cyano- N -(2-(l-(quinolin-8-ylmethyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-yl)acetamide

[0223]

[0224] The preparation method refers to Example 1, except that benzyl bromide is replaced by 6-(bromomethyl)quinoxaline to prepare 2-cyano- N -(2-(l-(quinolin-8-ylmethyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-yl)acetamide as a white solid (yield 71.3%). 1H NMR (300 MHz, DMSO- d 6 ) δ 8.99–8.86 (m, 2H), 8.69 (t, J =5.5 Hz, 1H), 8.28 (dd, J = 8.1, 1.5 Hz, 1H), 8.06 (d, J = 8.6 Hz, 1H), 7.96 (d, J =1.8 Hz, 1H), 7.82 (dd, J = 8.6, 1.9 Hz, 1H), 7.72-7.58 (m, 2H), 7.62-7.53 (m,1H), 7.51 (dd, J = 8.4, 1.2 Hz, 1H), 7.43 (td, J = 7.8, 7.4, 1.2 Hz, 2H), 7.26(ddd, J = 8.3, 7.2, 1.3 Hz, 1H), 3.68 (s, 2H), 3.13 (q, J = 6.6 Hz, 2H), 2.80(d, J = 10.7 Hz, 2H), 1.90 (t, J = 10.1 Hz, 2H), 1.56 (d, J = 9.6 Hz, 2H), 1.29 (q, J = 6.6 Hz, 2H), 1.19-0.98 (m, 3H). HRMS (ESI) m / z: [M + H] + calcd forC 32 H 30 N5O2, 516.2400; found, 516.2402.

[0225] Example 39

[0226] N -(2-(1-(benzo[d]oxazol-6-ylmethyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -Xanthan-9-ylidene)acetamide

[0227]

[0228] The preparation method is the same as in Example 1, except that benzyl bromide is replaced with 6-(bromomethyl)benzo[d ]oxazole, prepared N -(2-(1-(benzo[d]oxazol-6-ylmethyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (73.4% yield). 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.77-8.62 (m, 2H), 8.28(dd, J = 8.1, 1.5 Hz, 1H), 7.73 (d, J = 8.1 Hz, 1H), 7.70-7.60 (m, 3H), 7.60-7.54(m, 1H), 7.51 (dd, J = 8.4, 1.2 Hz, 1H), 7.47-7.38 (m, 2H), 7.33 (dd, J = 8.2,1.4 Hz, 1H), 7.30-7.18 (m, 1H), 3.56 (s, 2H), 3.12 (q, J = 6.6 Hz, 2H), 2.76(d, J = 10.8 Hz, 2H), 1.83 (t, J = 10.6 Hz, 2H), 1.54 (d, J = 9.2 Hz, 2H), 1.28(q, J = 6.5 Hz, 2H), 1.17-0.95 (m, 3H). HRMS (ESI) m / z: [M + H] + calcd forC 31 H 29 N4O3, 505.2240; found,505.2243.

[0229] Example 40

[0230] N -(2-(1-((6-bromobenzo[ d ][1,3]dioxolan-5-yl)methyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -xanthen-9-ylidene)acetamide

[0231]

[0232] The preparation method refers to Example 1, except that benzyl bromide is replaced by 5-bromo-6-(bromomethyl)benzo[ d ][1,3]dioxolane to produce N -(2-(1-((6-bromo d ][1,3]dioxol-5-yl)methyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (yield 63.4%). 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.68 (t, J = 5.5 Hz, 1H), 8.29 (dd, J = 8.1, 1.5 Hz, 1H), 7.72-7.62 (m,2H), 7.62-7.56 (m, 1H), 7.54-7.49 (m, 1H), 7.48-7.39 (m, 2H), 7.32-7.23 (m,1H), 7.16 (s, 1H), 6.99 (s, 1H), 6.06 (s, 2H), 3.39 (s, 2H), 3.13 (q, J = 6.6Hz, 2H), 2.76 (d, J = 10.8 Hz, 2H), 2.04-1.79 (m, 2H), 1.64-1.43 (m, 2H), 1.37-1.21 (m, 2H), 1.18-0.88 (m, 3H). HRMS (ESI) m / z: [M + H] + calcd for C 31 H 29 N3O4Br, 586.1341; found,586.1346.

[0233] Example 41

[0234] N -(2-(1-(benzofuran-2-ylmethyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -xanthen-9-ylidene)acetamide

[0235]

[0236] The preparation method refers to Example 1, except that benzyl bromide is replaced by 2-(bromomethyl)benzofuran to produce N- (2-( 1 -(benzofuran-2-ylmethyl)piperidin-4-yl)ethyl)-2-(9 H - xanthen-9-yl)acetamide as a white solid (yield 52.8%). 1 H NMR (300 MHz, DMSO-d6) δ 8.36 (t, J = 5.5 Hz, 1H), 8.05-7.88 (m, 1H), 7.41-7.31 (m, 2H), 7.31-7.24 (m, 2H), 7.24-7.17 (m, 2H), 7.16-7.04 (m, 2H), 7.02-6.85 (m, 3H), 6.44 (s, 1H), 3.31 (s, 2H), 2.80 (q, J =6.5 Hz, 2H), 2.51 (d, J = 11.0 Hz, 2H), 1.60 (t, J = 10.6 Hz, 2H), 1.24 (d, J =10.9 Hz, 2H), 0.96 (q, J = 6.7 Hz, 2H), 0.86-0.61 (m, 3H). HRMS (ESI) m / z: [M+ H] + calcd for C 32 H 30 N3O3, 504.2287; found, 504.2292.

[0237] Example 42

[0238] 2-cyano- N - (2-( 1 -(benzofuran-2-ylmethyl)piperidin-4-yl)ethyl)-2-(9 H - xanthen-9-yl)acetamide

[0239]

[0240] The preparation method refers to Example 1, except that benzyl bromide is replaced by 2-bromo-l-(4-methoxyphenyl)ethanone to prepare 2-cyano- N - (2-( 1 -(benzofuran-2-ylmethyl)piperidin-4-yl)ethyl)-2-(9 H - xanthen-9-yl)acetamide as a white solid (yield 52.8%). 1H NMR (300 MHz, DMSO-d6) δ 8.69 (t, J =5.5 Hz, 1H), 8.35-8.21 (m, 1H), 7.99 (d, J = 8.9 Hz, 2H), 7.72-7.62 (m, 2H), 7.62-7.56 (m, 1H), 7.55-7.39 (m, 3H), 7.34-7.22 (m, 1H), 7.03 (d, J = 8.9 Hz,2H), 3.84 (s, 3H), 3.67 (s, 2H), 3.12 (q, J = 6.6 Hz, 2H), 2.82 (d, J = 10.6 Hz,2H), 2.08-1.83 (m, 2H), 1.61-1.43 (m, 2H), 1.27 (q, J = 7.2, 6.7 Hz, 2H), 1.15-0.94 (m, 3H). HRMS (ESI) m / z: [M + Na] + calcd for C 32 H 31 N3O4Na, 544.2212; found, 544.2209.

[0241] Example 43

[0242] N -(2-(1-([1,1'-biphenyl]-4-ylmethyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -Xanthan-9-ylidene)acetamide

[0243]

[0244] The preparation method is the same as in Example 1, except that benzyl bromide is replaced with 4-(bromomethyl)-1,1'-biphenyl to obtain... N -(2-(1-([1,1'-biphenyl]-4-ylmethyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -Xanton-9-ylidene)acetamide, a white solid (yield 55.6%). 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.68 (t, J= 5.5 Hz, 1H), 8.29 (dd, J = 8.1, 1.5 Hz, 1H), 7.75-7.55 (m, 7H), 7.54-7.42 (m, 5H), 7.42-7.31(m, 3H), 7.31-7.18 (m, 1H), 3.62-3.47 (m, 2H), 3.13 (q, J = 6.6 Hz, 2H), 2.80(d, J = 10.8 Hz, 2H), 2.03-1.73 (m, 2H), 1.57 (d, J = 9.2 Hz, 2H), 1.29 (q, J =6.8, 6.4 Hz, 2H), 1.19-0.94 (m, 3H). HRMS (ESI) m / z: [M + H] + calcd forC 36 H 34 N3O2, 540.2651; found, 540.2656.

[0245] Example 44

[0246] N -(2-(1-(4-(benzyloxy)benzyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -Xanthan-9-ylidene)acetamide

[0247]

[0248] The preparation method is the same as in Example 23, except that p-hydroxybenzaldehyde is replaced with 4-(benzyloxy)benzaldehyde to obtain... N -(2-(1-(4-(benzyloxy)benzyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -Xanton-9-ylidene)acetamide, a white solid (yield 78.6%). 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.68 (t, J = 5.5 Hz, 1H), 8.29 (dd, J = 8.1, 1.5 Hz, 1H), 7.72-7.60 (m, 2H), 7.60-7.55 (m, 1H), 7.51(dd, J= 8.4, 1.2 Hz, 1H), 7.48-7.41 (m, 4H), 7.41-7.39 (m, 1H), 7.38-7.30 (m,2H), 7.29-7.22 (m, 1H), 7.21-7.09 (m, 2H), 6.97 (d, J = 8.1 Hz, 2H), 5.09 (s,2H), 3.54-3.36 (m, 2H), 3.12 (q, J = 6.7 Hz, 2H), 2.90-2.61 (m, 2H), 2.07-1.69(m, 2H), 1.64-1.47 (m, 2H), 1.35-1.17 (m, 2H), 1.15-0.95 (m, 3H). HRMS (ESI)m / z: [M + H] + calcd for C 37 H 36 N3O3, 570.2757; found, 570.2761.

[0249] Example 45

[0250] N -(2-(1-(3-(benzyloxy)benzyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -Xanthan-9-ylidene)acetamide

[0251]

[0252] The preparation method is the same as in Example 23, except that p-hydroxybenzaldehyde is replaced with 3-(benzyloxy)benzaldehyde to obtain... N -(2-(1-(3-(benzyloxy)benzyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -Xanton-9-ylidene)acetamide, a white solid (yield 74.2%). 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.67 (t, J = 5.5 Hz, 1H), 8.29 (dd, J = 8.1, 1.5 Hz, 1H), 7.73-7.58 (m, 2H), 7.62-7.53 (m, 1H), 7.51 (dd, J = 8.4, 1.2 Hz, 1H), 7.50-7.27 (m, 7H), 7.24 (dt,J = 10.0, 7.9 Hz, 2H), 6.99-6.80 (m, 3H), 5.09 (s, 2H), 3.39 (s, 2H), 3.12 (q, J = 6.6 Hz, 2H), 2.72 (d, J =10.7 Hz, 2H), 1.91-1.66 (m, 2H), 1.53 (d, J = 8.3 Hz, 2H), 1.28 (q, J = 6.5 Hz,2H), 1.15-0.93 (m, 3H). HRMS (ESI) m / z: [M + H] + calcd for C 37 H 36 N3O3, 570.2757;found,570.2759。

[0253] Example 46

[0254] N -(2-(1-(2-(benzyloxy)benzyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -xanthen-9-ylidene)acetamide

[0255]

[0256] Preparation method according to example 23, except that 2-(benzyloxy)benzaldehyde is used instead of p-hydroxybenzaldehyde, affords N -(2-(1-(2-(benzyloxy)benzyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (77.6% yield). 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.67 (t, J = 5.6 Hz, 1H),8.29 (dd, J = 8.1, 1.5 Hz, 1H), 7.73-7.60 (m, 2H), 7.61-7.44 (m, 4H), 7.48-7.35(m, 4H), 7.32 (t, J = 7.1 Hz, 2H), 7.23 (q, J = 7.4 Hz, 2H), 7.06 (d, J= 8.2 Hz, 1H), 6.93 (t, J = 7.4 Hz, 1H), 5.13 (s, 2H), 3.74-3.42 (m, 2H), 3.12 (q, J = 6.5Hz, 2H), 2.98-2.62 (m, 2H), 2.09-1.69 (m, 2H), 1.56 (d, J = 9.5 Hz, 2H), 1.28(q, J = 6.6 Hz, 2H), 1.18-0.94 (m, 3H). HRMS (ESI) m / z: [M + H] + calcd forC 37 H 36 N3O3, 570.2757; found, 570.2760.

[0257] Example 47

[0258] 2-Cyano- N -(2-(1-(4-(phenoxymethyl)benzyl)piperidin-4-yl)ethyl)-2-(9 H -Xanthan-9-ylidene)acetamide

[0259]

[0260] The preparation method is the same as in Example 23, except that p-hydroxybenzaldehyde is replaced with 4-(phenoxymethyl)benzaldehyde to obtain... N -(2-(1-(2-(benzyloxy)benzyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -Xanton-9-ylidene)acetamide, a white solid (yield 69.3%). 1 H NMR (600 MHz, Chloroform- d ) δ 8.39 (dd, J =8.1, 1.5 Hz, 1H), 7.68 (dd, J = 8.1, 1.5 Hz, 1H), 7.53-7.46 (m, 2H), 7.46-7.38(m, 4H), 7.35-7.27 (m, 5H), 7.19-7.11 (m, 1H), 7.02-6.91 (m, 3H), 6.12 (s,1H), 5.05 (s, 2H), 3.75 (s, 2H), 3.31 (q, J= 6.7 Hz, 2H), 3.08 (d, J = 11.5 Hz,2H), 2.28 (t, J = 11.0 Hz, 2H), 1.73 (d, J = 14.2 Hz, 2H), 1.48-1.43 (m, 4H),1.39-1.29 (m, 1H). HRMS (ESI) m / z: [M + H] + calcd for C 37 H 36 N3O3, 570.2751;found,570.2753.

[0261] Example 48

[0262] 2-cyano- N -(2-(1-(4-((4-fluorobenzyl)oxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0263]

[0264] Preparation method according to example 23, except that 4-((4- fluorobenzyl)oxy)benzaldehyde is used instead of p-hydroxybenzaldehyde to give 2-cyano- N -(2-(1-(4-((4-fluorobenzyl)oxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (yield 70.5%). 1 H NMR (300 MHz, Chloroform- d ) δ 8.40(d, J = 8.0 Hz, 1H), 7.71 (d, J = 8.0 Hz, 1H), 7.60-7.44 (m, 2H), 7.44-7.28 (m,5H), 7.26-7.20 (m, 2H), 7.20-7.00 (m, 3H), 6.91 (d, J = 8.1 Hz, 2H), 5.89 (t, J =5.8 Hz, 1H), 5.01 (s, 2H), 3.47 (s, 2H), 3.32 (q, J = 6.8 Hz, 2H), 2.89 (d, J= 11.3 Hz, 2H), 2.06-1.88 (m, 2H), 1.63 (d, J = 10.9 Hz, 2H), 1.41 (q, J = 6.7 Hz,2H), 1.36-1.17 (m, 3H). HRMS (ESI) m / z: [M + H] + calcd for C 37 H 35 FN3O3,588.2657; found,588.2656.

[0265] Example 49

[0266] 2-cyano- N -(2-(1-(4-((4-(trifluoromethoxy)benzyl)oxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0267]

[0268] Preparation method according to example 23, except that 4-((4- (trifluoromethoxy)benzyl)oxy)benzaldehyde is used instead of p-hydroxybenzaldehyde, to produce 2-cyano- N -(2-(1-(4-((4-(trifluoromethoxy)benzyl)oxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (76.5% yield). 1 H NMR (300MHz, Chloroform- d ) δ 8.41 (d, J = 8.1 Hz, 1H), 7.72 (d, J = 8.0 Hz, 1H), 7.59-7.41(m, 4H), 7.40-7.28 (m, 3H), 7.26-7.19 (m, 4H), 7.15 (t, J = 7.6 Hz, 1H), 7.04-6.82 (m, 2H), 5.83 (t, J = 5.8 Hz, 1H), 5.04 (s, 2H), 3.46 (s, 2H), 3.32 (q, J =6.8 Hz, 2H), 2.87 (d, J = 11.1 Hz, 2H), 1.92 (t,J = 10.9 Hz, 2H), 1.62 (d, J =11.0 Hz, 2H), 1.41 (q, J = 6.7 Hz, 2H), 1.34-1.19 (m, 3H). HRMS (ESI) m / z: [M +H] + calcd for C 38 H 36 N3O4F2, 654.2574; found,654.2573.

[0269] Example 50

[0270] 2-cyano- N -(2-(1-(4-((4-(di- fluoromethoxy)benzyl)oxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0271]

[0272] Preparation method according to example 23, except that p-hydroxybenzaldehyde is replaced by 4-((4-(di-fluoromethoxy)benzyl)oxy)benzaldehyde, to produce 2-cyano- N -(2-(1-(4-((4-(di- fluoromethoxy)benzyl)oxy)benzyl)piperidin-4-yl)ethyl)-2-(9H-xanthen-9-ylidene)acetamide as a white solid (yield 78.6%). 1 H NMR (400MHz, Chloroform- d ) δ 8.43 (dd, J = 8.1, 1.5 Hz, 1H), 7.73 (dd, J = 8.1, 1.5 Hz,1H), 7.59-7.53 (m, 1H), 7.53-7.48 (m, 1H), 7.48-7.42 (m, 2H), 7.40-7.31 (m,3H), 7.28-7.23 (m, 2H), 7.21-7.12 (m, 3H), 6.94 (d, J = 8.6 Hz, 2H), 6.53 (t, J =73.9 Hz, 1H), 5.92 (t, J= 5.8 Hz, 1H), 5.05 (s, 2H), 3.50 (s, 2H), 3.41-3.27(m, 2H), 2.92 (d, J = 11.4 Hz, 2H), 1.97 (t, J = 11.2 Hz, 2H), 1.71-1.60 (m,2H), 1.43 (q, J = 7.0 Hz, 2H), 1.36-1.17 (m, 3H). HRMS (ESI) m / z: [M + H] + calcd for C 38 H 36 N3O4F2, 636.2674; found, 636.2676.

[0273] Example 51

[0274] N -(2-(1-(3-(benzylamino)benzyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -Xanton-9-ylidene)acetamide

[0275]

[0276] The preparation method is the same as in Example 23, except that p-hydroxybenzaldehyde is replaced with 3-(benzylamino)benzaldehyde to obtain... N -(2-(1-(3-(benzylamino)benzyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -Xanton-9-ylidene)acetamide, a white solid (yield 58.1%). 1 H NMR (300 MHz, Chloroform- d ) δ 8.42 (dd, J = 8.1,1.5 Hz, 1H), 7.80-7.66 (m, 1H), 7.63-7.46 (m, 2H), 7.44-7.27 (m, 8H), 7.22-7.10 (m, 2H), 6.76 (t, J = 2.0 Hz, 1H), 6.71-6.56 (m, 2H), 6.19 (t, J = 5.8 Hz,1H), 4.36 (s, 2H), 3.70 (s, 2H), 3.33 (q, J = 6.7 Hz, 2H), 3.09 (d,J = 11.6 Hz,2H), 2.34-2.11 (m, 2H), 1.79-1.61 (m, 2H), 1.62-1.54 (m, 1H), 1.46 (q, J = 7.0Hz, 2H), 1.38-1.21 (m, 2H). HRMS (ESI) m / z: [M + H] + calcd for C 37 H 37 N4O2,569.2917; found,569.2919。

[0277] Example 52

[0278] 2-cyano- N -(2-(1-(4-(pyridin-4-ylmethoxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0279]

[0280] Preparation method according to Example 23, except that 4-(pyridin-4- ylmethoxy)benzaldehyde is used instead of p-hydroxybenzaldehyde to produce 2-cyano- N -(2-(1-(4-(pyridin-4-ylmethoxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (yield 61.7%). 1 H NMR (300 MHz, Chloroform- d ) δ 8.67-8.53 (m, 2H), 8.40 (d, J = 7.9 Hz, 1H), 7.80-7.66 (m, 1H), 7.62-7.43 (m,2H), 7.41-7.29 (m, 5H), 7.28-7.21 (m, 2H), 7.15 (t, J = 7.6 Hz, 1H), 6.97-6.83(m, 2H), 6.19-6.07 (m, 1H), 5.08 (s, 2H), 3.51 (s, 2H), 3.32 (q, J = 6.8 Hz,2H), 2.97-2.89 (m, 2H), 2.07-1.96 (m, 2H), 1.64 (d, J= 11.6 Hz, 2H), 1.42 (q, J = 7.5, 7.1 Hz, 2H), 1.35-1.18 (m, 3H). HRMS (ESI) m / z: [M + H] + calcd forC 36 H 35 N4O3, 570.2704; found,571.2707.

[0281] Example 53

[0282] 2-cyano- N -(2-(l-(4-(pyrimidin-2-ylmethoxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0283]

[0284] Preparation Method refers to Example 23, except that p-hydroxybenzaldehyde is replaced by 4-(pyrimidin-2-ylmethoxy)benzaldehyde, 2-cyano- N -(2-(l-(4-(pyrimidin-2-ylmethoxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide is prepared as a white solid (yield 64.6%). 1 H NMR (400 MHz, Chloroform- d ) δ 8.80 (d, J = 4.9 Hz, 2H), 8.42 (dd, J = 8.1, 1.5 Hz, 1H), 7.73 (dd, J = 8.0, 1.5Hz, 1H), 7.61-7.45 (m, 2H), 7.41-7.30 (m, 3H), 7.28-7.21 (m, 3H), 7.20-7.13(m, 1H), 7.03-6.93 (m, 2H), 5.96 (t, J = 5.7 Hz, 1H), 5.31 (s, 2H), 3.51 (s,2H), 3.40-3.26 (m, 2H), 3.00-2.87 (m, 2H), 2.04-1.94 (m, 2H), 1.73-1.57 (m,2H), 1.43 (q, J= 6.9 Hz, 2H), 1.38-1.23 (m, 3H). HRMS (ESI) m / z: [M + H] + calcdfor C 35 H 34 N5O3, 572.2656; found,571.2656.

[0285] Example 54

[0286] 2-cyano- N -(2-(1-(4-(cyclohexylmethoxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0287]

[0288] Preparation method according to example 23, except that 4-(cyclohexylmethoxy)benzaldehyde is used instead of p-hydroxybenzaldehyde to produce 2-cyano- N -(2-(1-(4-(cyclohexylmethoxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (yield 70.5%). 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.67 (t, J = 5.6 Hz, 1H), 8.28 (d, J = 8.0 Hz, 1H), 7.77-7.60 (m, 2H), 7.57 (d, J = 7.6 Hz,1H), 7.51 (d, J = 8.2 Hz, 1H), 7.48-7.35 (m, 2H), 7.25 (t, J = 7.6 Hz, 1H), 7.15(d, J = 8.1 Hz, 2H), 6.84 (d, J = 8.0 Hz, 2H), 3.73 (d, J = 6.1 Hz, 2H), 3.32 (s,2H), 3.11 (q, J = 6.5 Hz, 2H), 2.71 (d, J = 10.8 Hz, 2H), 1.86-1.65 (m, 8H), 1.53(d, J= 8.4 Hz, 2H), 1.33-1.17 (m, 5H), 1.12-0.92 (m, 5H). HRMS (ESI) m / z: [M +H] + calcd for C 37 H 42 N3O3, 576.3221; found,576.3227.

[0289] Example 55

[0290] 2-cyano- N -(2-(1-(4-(cyclopropylmethoxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0291]

[0292] The preparation method refers to Example 23, except that p-hydroxybenzaldehyde is replaced by 4-(cyclopropylmethoxy)benzaldehyde to prepare 2-cyano- N -(2-(1-(4-(cyclopropylmethoxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (yield 71.1%). 1 H NMR (300 MHz, Chloroform-d) δ 8.40 (d, J = 8.0 Hz, 1H), 7.71 (d, J = 7.9 Hz, 1H), 7.50 (dt, J = 14.5, 7.8 Hz, 2H), 7.32 (q, J = 6.9 Hz, 3H), 7.22 (d, J = 8.2 Hz, 2H), 7.14 (t, J = 7.7 Hz, 1H), 6.85 (d, J = 8.1 Hz, 2H), 5.94 (d, J = 6.0 Hz, 1H), 3.79 (d, J = 6.9 Hz, 2H), 3.51 (s, 2H), 3.31 (q, J = 6.7 Hz, 2H), 2.92 (d, J = 11.2 Hz, 2H), 1.99 (t, J= 11.5 Hz,2H), 1.64 (d, J = 12.3 Hz, 2H), 1.41 (q, J = 6.8 Hz, 2H), 1.38-1.12 (m, 4H), 0.64(q, J = 5.6 Hz, 2H), 0.43-0.25 (m, 2H). HRMS (ESI) m / z: [M + H] + calcd forC 34 H 36 N3O3, 534.2757; found,534.2759。

[0293] Example 56

[0294] 2-cyano- N -(2-(1-(4-((tetrahydro-2 H -pyran-4-yl)methoxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0295]

[0296] The preparation method refers to Example 23, except that p-hydroxybenzaldehyde is replaced by 4-((tetrahydro-2 H -pyran-4-yl)methoxy)benzaldehyde to produce 2-cyano- N -(2-(1-(4-((tetrahydro-2 H -pyran-4-yl)methoxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (yield 72.1%). 1 H NMR (300MHz, DMSO- d 6 ) δ 8.66 (t, J = 5.6 Hz, 1H), 8.28 (d, J = 8.0 Hz, 1H), 7.65 (q, J = 8.4Hz, 2H), 7.57 (d, J = 7.6 Hz, 1H), 7.51 (d, J = 8.2 Hz, 1H), 7.47-7.36 (m, 2H),7.25 (t, J = 7.6 Hz, 1H), 7.17 (d, J= 8.1 Hz, 2H), 6.87 (d, J = 8.0 Hz, 2H), 3.94-3.72 (m, 4H), 3.42-3.32 (m, 4H), 3.11 (q, J = 6.6 Hz, 2H), 2.73 (d, J = 10.3 Hz,2H), 2.09-1.90 (m, 1H), 1.89-1.72 (m, 2H), 1.72-1.61 (m, 2H), 1.60-1.46 (m,2H), 1.36-1.22 (m, 4H), 1.14-0.97 (m, 3H). HRMS (ESI) m / z: [M + H] + calcd forC 36 H 40 N3O4, 578.3013; found,576.3015。

[0297] Example 57

[0298] tert-Butyl 4-((4-((4-(2-(2-cyano-2-(9 H -xanthen-9-ylidene)acetamido)ethyl)piperidin-1-yl)methyl)phenoxy)methyl)piperidine-1-carboxylate

[0299]

[0300] Preparation Method refers to Example 23, except that p-hydroxybenzaldehyde is replaced by tert-butyl 4-((4-formylphenoxy)methyl)piperidine-1-carboxylate, to prepare tert-butyl 4-((4-((4-(2-(2-cyano-2-(9 H -xanthen-9-ylidene)acetamido)ethyl)piperidin-1-yl)methyl)phenoxy)methyl)piperidine-1-carboxylate, as a white solid (yield 54.3%). 1 H NMR (300 MHz, Chloroform- d ) δ 8.47-8.35 (m, 1H), 7.79-7.66 (m, 1H),7.60-7.42 (m, 2H), 7.39-7.26 (m, 3H), 7.26-7.09 (m, 3H), 6.90-6.75 (m, 2H),6.01-5.71 (m, 1H), 4.35-3.98 (m, 2H), 3.78 (d, J= 6.3 Hz, 2H), 3.46 (s, 2H),3.32 (q, J = 6.8 Hz, 2H), 2.88 (d, J = 11.2 Hz, 2H), 2.74 (t, J = 12.9 Hz, 2H),1.92 (t, J = 10.3 Hz, 3H), 1.81 (d, J = 13.1 Hz, 2H), 1.62 (d, J = 11.2 Hz, 2H),1.46 (s, 9H), 1.43-1.35 (m, 1H), 1.35-1.12 (m, 6H). HRMS (ESI) m / z: [M + H] + calcd for C 41 H 49 N4O5, 677.3697; found,677.3698.

[0301] Example 58

[0302] 4-((4-(2-(2-cyano-2-(9 H -xanthen-9-ylidene)acetamido)ethyl)piperidin-1-yl)methyl)phenyl 2-(adamantan-1- yl)acetate

[0303]

[0304] Preparation method according to example 23, except that 4-formylphenyl 2-(adamantan-1- yl)acetate is used instead of 4-hydroxybenzaldehyde to give 4-((4-(2-(2-cyano-2-(9 H -xanthen-9-ylidene)acetamido)ethyl)piperidin-1-yl)methyl)phenyl 2-(adamantan-1- yl)acetate as a white solid (59.1% yield). 1 H NMR (600MHz, Chloroform- d ) δ 8.45-8.37 (m, 1H), 7.76-7.70 (m, 1H), 7.58-7.51 (m, 1H),7.51-7.45 (m, 1H), 7.38-7.29 (m, 5H), 7.15 (t, J = 7.6 Hz, 1H), 7.03 (d, J = 8.2Hz, 2H), 5.84 (t, J= 5.7 Hz, 1H), 3.51 (s, 2H), 3.32 (q, J = 6.6 Hz, 2H), 2.94-2.83 (m, 2H), 2.29 (s, 2H), 2.03-1.99 (m, 3H), 1.99-1.89 (m, 2H), 1.76-1.71(m, 9H), 1.70-1.65 (m, 3H), 1.62 (d, J = 12.4 Hz, 2H), 1.42 (q, J = 7.1 Hz, 2H),1.37-1.27 (m, 2H), 1.24-1.17 (m, 1H). HRMS (ESI) m / z: [M + H] + calcd for C 42 H 46 N3O4, 656.3483; found,677.3487.

[0305] Example 59

[0306] 2-cyano- N -(2-(1-(4-(2-(4-methoxyphenyl)-2-oxoethoxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0307]

[0308] The procedure of Reference Example 23 was followed except that 4-(2-(4- methoxyphenyl)-2-oxoethoxy)benzaldehyde was used instead of p-hydroxybenzaldehyde to give 2-cyano- N -(2-(1-(4-(2-(4-methoxyphenyl)-2-oxoethoxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (64.5% yield). 1 HNMR (300 MHz, Chloroform- d ) δ 8.40 (d, J = 8.0 Hz, 1H), 7.98 (d, J = 8.5 Hz, 2H),7.71 (d, J= 8.0 Hz, 1H), 7.60-7.41 (m, 2H), 7.39-7.27 (m, 3H), 7.26-7.09 (m,3H), 7.02-6.78 (m, 4H), 5.96 (t, J = 5.5 Hz, 1H), 5.19 (s, 2H), 3.88 (s, 3H),3.45 (s, 2H), 3.31 (q, J = 6.8 Hz, 2H), 2.86 (d, J = 11.0 Hz, 2H), 1.92 (t, J =11.1 Hz, 2H), 1.61 (d, J = 11.4 Hz, 2H), 1.40 (q, J = 6.7 Hz, 2H), 1.34-1.21 (m,3H). HRMS (ESI) m / z: [M + H] + calcd for C 39 H 38 N3O5, 628.2806; found,628.2810.

[0309] Example 60

[0310] 2-cyano- N -(2-(1-(4-phenethoxybenzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0311]

[0312] Preparation method according to example 23, except that p-hydroxybenzaldehyde is replaced by 4-phenethoxybenzaldehyde, to produce 2-cyano- N -(2-(1-(4-phenethoxybenzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (yield 81.6%). 1 H NMR (400 MHz, Chloroform- d ) δ 8.42 (dd, J = 8.1,1.5 Hz, 1H), 7.72 (dd, J= 8.1, 1.5 Hz, 1H), 7.61-7.45 (m, 2H), 7.39-7.30 (m,6H), 7.30-7.22 (m, 4H), 7.22-7.12 (m, 1H), 6.92-6.79 (m, 2H), 6.04 (t, J = 5.9Hz, 1H), 4.18 (t, J = 7.1 Hz, 2H), 3.59 (s, 2H), 3.33 (q, J = 6.8 Hz, 2H), 3.11(t, J = 7.1 Hz, 2H), 2.98 (d, J = 11.3 Hz, 2H), 2.18-1.93 (m, 2H), 1.74-1.60 (m,2H), 1.52-1.36 (m, 4H), 1.32-1.22 (m, 1H). HRMS (ESI) m / z: [M + H] + calcd for C 38 H 38 N3O3, 584.2908; found,584.2909.

[0313] Example 61

[0314] 2-cyano- N -(2-(1-(4-(2-fluorophenethoxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0315]

[0316] The preparation method refers to Example 23, except that p-hydroxybenzaldehyde is replaced by 4-(2-fluorophenethoxy)benzaldehyde to prepare 2-cyano- N -(2-(1-((4-(2-fluorophenethoxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide, a white solid (yield 73.2%). 1 H NMR (400 MHz, Chloroform- d ) δ 8.42(dd, J = 8.1, 1.5 Hz, 1H), 7.73 (dd, J= 8.1, 1.5 Hz, 1H), 7.60-.46 (m, 2H),7.40-7.29 (m, 4H), 7.28-7.20 (m, 3H), 7.20-7.14 (m, 1H), 7.14-7.02 (m, 2H),6.92-6.84 (m, 2H), 5.95 (t, J = 5.8 Hz, 1H), 4.19 (t, J = 7.0 Hz, 2H), 3.56 (s,2H), 3.40-3.28 (m, 2H), 3.15 (t, J = 7.0 Hz, 2H), 2.96 (d, J = 11.3 Hz, 2H),2.12-1.94 (m, 2H), 1.74-1.59 (m, 2H), 1.44 (q, J = 6.9 Hz, 2H), 1.40-1.31 (m,2H), 1.32-1.23 (m, 1H). HRMS (ESI) m / z: [M + H] + calcd for C 38 H 37 FN3O3, 602.2813; found, 602.2815.

[0317] Example 62

[0318] 2-cyano- N -(2-(1-(4-(2-bromophenethoxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0319]

[0320] The preparation method refers to Example 23, except that p-hydroxybenzaldehyde is replaced by 4-(2-bromophenethoxy)benzaldehyde to prepare 2-cyano- N -(2-(1-((4-(2-bromophenethoxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide, which is a white solid (yield 70.5%). 1 H NMR (400 MHz, Chloroform- d ) δ 8.43(dd, J = 8.1, 1.5 Hz, 1H), 7.73 (dd, J= 8.1, 1.5 Hz, 1H), 7.60-7.47 (m, 3H),7.39-7.29 (m, 4H), 7.29-7.26 (m, 1H), 7.26-7.20 (m, 2H), 7.20-7.09 (m, 2H),6.91-6.85 (m, 2H), 5.89 (t, J = 5.7 Hz, 1H), 4.20 (t, J = 7.0 Hz, 2H), 3.50 (s,2H), 3.38-3.30 (m, 2H), 3.26 (t, J = 7.0 Hz, 2H), 2.96-2.87 (m, 2H), 1.96 (t, J =11.9 Hz, 2H), 1.70-1.60 (m, 2H), 1.43 (q, J = 7.0 Hz, 2H), 1.37-1.22 (m, 3H).HRMS (ESI) m / z: [M + H] + calcd for C 38 H 37 N3O3Br, 662.2018; found,662.2015.

[0321] Example 63

[0322] 2-cyano- N -(2-(1-(4-(2-(pyridin-2-yl)ethoxy)benzyl]piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0323]

[0324] The preparation method refers to Example 23, except that p-hydroxybenzaldehyde is replaced by 4-(2-(pyridin-2-yl)ethoxy)benzaldehyde to obtain 2-cyano- N -(2-(1-(4-(2-(pyridin-2-yl)ethoxy)benzyl]piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (yield 66.7%). 1 H NMR (300 MHz, Chloroform- d ) δ 8.53 (d, J = 4.9 Hz, 1H), 8.40 (d, J= 8.1 Hz, 1H), 7.71 (d, J =8.0 Hz, 1H), 7.67-7.58 (m, 1H), 7.57-7.42 (m, 2H), 7.40-7.29 (m, 3H), 7.29-7.26 (m, 1H), 7.24-7.18 (m, 2H), 7.18-7.08 (m, 2H), 6.86 (d, J = 8.1 Hz, 2H), 6.05 (t, J = 5.7 Hz, 1H), 4.35 (t, J = 6.6 Hz, 2H), 3.51 (s, 2H), 3.39-3.16 (m,4H), 2.92 (d, J = 11.6 Hz, 2H), 1.99 (t, J = 10.8 Hz, 2H), 1.63 (d, J = 12.1 Hz,2H), 1.47-1.38 (m, 2H), 1.35-1.27 (m, 3H). HRMS (ESI) m / z: [M + H] + calcd forC 37 H 37 N4O3, 585.2860; found, 585.2862.

[0325] Example 64

[0326] 2-Cyano- N -(2-(1-(4-(2-morpholinoethoxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -Xanthan-9-ylidene)acetamide

[0327]

[0328] The preparation method is the same as in Example 23, except that p-hydroxybenzaldehyde is replaced with 4-(2-morpholinoethoxy)benzaldehyde to obtain 2-cyano- N -(2-(1-(4-(2-morpholinoethoxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -Xanton-9-ylidene)acetamide, a white solid (yield 69.4%). 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.67 (t, J=5.4 Hz, 1H), 8.28 (d, J = 8.0 Hz, 1H), 7.80-7.60 (m, 2H), 7.57 (d, J = 7.7 Hz, 1H), 7.51 (d, J = 8.2 Hz, 1H), 7.48-7.38 (m, 2H), 7.30-7.11 (m, 3H), 6.90 (d, J =8.0 Hz, 2H), 4.06 (t, J = 5.8 Hz, 2H), 3.57 (t, J = 4.6 Hz, 5H), 3.48-3.35 (m,2H), 3.11 (q, J = 6.5 Hz, 2H), 2.95-2.73 (m, 2H), 2.68 (t, J = 5.7 Hz, 2H), 2.48-2.37 (m, 3H), 2.12-1.71 (m, 1H), 1.67-1.46 (m, 2H), 1.37-1.19 (m, 3H), 1.19-0.93 (m, 3H). HRMS (ESI) m / z: [M + H] + calcd for C 36 H 41 N4O4, 593.3122; found, 593.3122.

[0329] Example 65

[0330] 2-Cyano- N -(2-(1-(4-(2-dimethylaminoethoxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -Xanthan-9-ylidene)acetamide

[0331]

[0332] The preparation method is the same as in Example 23, except that p-hydroxybenzaldehyde is replaced with 4-(2-morpholinoethoxy)benzaldehyde to obtain 2-cyano- N -(2-(1-(4-(2-dimethylaminoethoxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -Xanton-9-ylidene)acetamide, a white solid (yield 53.2%). 1 H NMR (300 MHz, Chloroform- d )δ 8.41 (dd, J = 8.1, 1.5 Hz, 1H), 7.72 (dd, J = 8.0, 1.5 Hz, 1H), 7.61-7.42 (m,2H), 7.39-7.26 (m, 3H), 7.25-7.09 (m, 3H), 6.92-6.80 (m, 2H), 5.89 (t, J = 6.0Hz, 1H), 4.05 (t, J = 5.7 Hz, 2H), 3.45 (s, 2H), 3.32 (q, J = 6.8 Hz, 2H), 2.87(d, J = 11.3 Hz, 2H), 2.73 (t, J = 5.7 Hz, 2H), 2.33 (s, 6H), 1.91 (t, J = 11.1 Hz,2H), 1.61 (d, J = 11.4 Hz, 2H), 1.41 (q, J = 6.9 Hz, 2H), 1.33-1.17 (m, 3H).HRMS (ESI) m / z: [M + H] + calcd for C 34 H 39 N4O3, 551.3017; found,551.3018。

[0333] Example 66

[0334] 2-cyano- N -(2-(1-(4-(2-piperidin-1-yl-ethoxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0335]

[0336] The preparation method refers to Example 23, except that p-hydroxybenzaldehyde is replaced by 4-(2-(piperidin-1-yl)ethoxy)benzaldehyde to produce 2-cyano- N -(2-(1-(4-(2-piperidin-1-yl-ethoxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (yield 55.4%). 1 H NMR (300 MHz, DMSO-d 6 ) δ 8.67 (t, J = 5.4 Hz, 1H), 8.28 (d, J = 8.0 Hz, 1H), 7.64 (q, J = 9.1, 8.7 Hz, 2H),7.56 (d, J = 7.7 Hz, 1H), 7.50 (d, J = 8.2 Hz, 1H), 7.47-7.38 (m, 2H), 7.31-7.10(m, 3H), 6.88 (d, J = 8.1 Hz, 2H), 4.05 (t, J = 5.8 Hz, 2H), 3.41 (s, 2H), 3.11(q, J = 6.4 Hz, 2H), 2.88-2.63 (m, 4H), 2.49-2.40 (m, 3H), 1.84 (t, J = 10.6 Hz,2H), 1.63-1.43 (m, 7H), 1.44-1.34 (m, 2H), 1.32-1.19 (m, 2H), 1.18-0.98 (m,3H). HRMS (ESI) m / z: [M + H] + calcd for C 37 H 43 N4O3, 591.3335; found,591.3339。

[0337] Example 67

[0338] 2-cyano- N -(2-(1- (4-(2- pyrrolidin-1-yl ethoxy) benzyl) piperidin-4-yl} ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0339]

[0340] The preparation method refers to Example 23, except that p-hydroxybenzaldehyde is replaced by 4-(2-(pyrrolidin-1-yl)ethoxy)benzaldehyde to prepare 2-cyano-N-(2-(1- (4-(2-pyrrolidin-1-yl ethoxy) benzyl) piperidin-4-yl} ethyl)-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (yield 59.8%). 1H NMR (300 MHz, DMSO- d 6 ) δ 8.68 (t, J = 5.5 Hz, 1H), 8.27 (d, J = 8.0 Hz, 1H), 7.73-7.59 (m, 2H), 7.56(d, J = 7.7 Hz, 1H), 7.50 (d, J = 8.2 Hz, 1H), 7.47-7.36 (m, 2H), 7.31-7.11 (m,3H), 6.89 (d, J = 8.1 Hz, 2H), 4.08 (t, J = 5.7 Hz, 2H), 3.41 (s, 2H), 3.18-3.04(m, 2H), 2.90 (t, J = 5.7 Hz, 2H), 2.76 (d, J = 11.1 Hz, 2H), 2.70-2.58 (m, 4H),1.95-1.77 (m, 2H), 1.77-1.65 (m, 4H), 1.62-1.46 (m, 2H), 1.33-1.19 (m, 2H),1.17-0.98 (m, 3H). HRMS (ESI) m / z: [M + H] + calcd for C 36 H 41 N4O3, 577.3179;found,577.3184。

[0341] Example 68

[0342] 2-cyano- N -(2-(l-(4-(2-phenoxyethoxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0343]

[0344] Preparation method according to Example 23, except that 4-(2-phenoxyethoxy)benzaldehyde is used instead of p-hydroxybenzaldehyde to give 2-cyano- N -(2-(l-(4-(2-phenoxyethoxy)benzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (54.7% yield).1 H NMR (400 MHz, Chloroform- d ) δ 8.43(dd, J = 8.1, 1.5 Hz, 1H), 7.74 (dd, J = 8.0, 1.5 Hz, 1H), 7.60-7.45 (m, 2H),7.40-7.29 (m, 5H), 7.27-7.22 (m, 2H), 7.21-7.13 (m, 1H), 7.03-6.88 (m, 5H),5.90 (t, J = 5.8 Hz, 1H), 4.34 (s, 4H), 3.47 (s, 2H), 3.39-3.27 (m, 2H), 2.88(d, J = 11.1 Hz, 2H), 1.92 (t, J = 11.0 Hz, 2H), 1.71-1.59 (m, 2H), 1.48-1.38 (m,2H), 1.32-1.20 (m, 3H). HRMS (ESI) m / z: [M + H] + calcd for C 38 H 38 N3O4, 600.2857;found,600.2856。

[0345] Example 69

[0346] 2-cyano- N -(2-(4-(4-(cyclohexylmethoxy)benzyl)piperazin-1-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0347]

[0348] The preparation method of Example 54, except that tert-butyl 4-(2- aminoethyl)piperazine-1-carboxylate is used to replace tert-butyl 4-(2- aminoethyl)piperidine-1-carboxylate in step (3), tert-butyl 4-(2-(2-cyano-2-(9 H -xanthen-9-ylidene)acetamido)ethyl)piperazine-1-carboxylate (d2) is used to replace tert-butyl 4-(2-(2-cyano-2-(9 H -xanthen-9-ylidene)acetamido)ethyl)piperidine-1-carboxylate in step (4), and 2-cyano- N-2-(9 H - xanthen-9-yl)acetamide to 2-cyano- N -2-(9 H - xanthen-9-yl)acetamide (e2) to 2-cyano- N -2-(9 H - xanthen-9-yl)acetamide as a white solid (yield 68.7%). 1 H NMR (300 MHz, Chloroform- d ) δ 8.44 (d, J = 8.0 Hz, 1H), 7.72 (d, J = 8.0 Hz, 1H), 7.55 (t, J = 7.8Hz, 1H), 7.46 (t, J = 7.7 Hz, 1H), 7.41-7.29 (m, 3H), 7.23-7.09 (m, 3H), 6.90-6.77 (m, 2H), 6.43 (t, J = 4.9 Hz, 1H), 3.74 (d, J = 6.1 Hz, 2H), 3.41 (s, 2H),3.36 (q, J = 5.5 Hz, 2H), 2.41 (t, J = 5.8 Hz, 3H), 2.33-2.25 (m, 5H), 1.94-1.82(m, 2H), 1.81-1.65 (m, 4H), 1.48-1.39 (m, 2H), 1.38-1.15 (m, 3H), 1.15-0.95(m, 2H). HRMS (ESI) m / z: [M + H] + calcd for C 36 H 41 N4O3, 577.3179; found,577.3184.

[0349] Example 70

[0350] 2-cyano- N -2-(9 H - xanthen-9-yl)acetamide

[0351]

[0352] The preparation method was referred to Example 54, except that tert-butyl 4-((2- cyano-2-(9 H -xanthen-9-yl)acetylamino)methyl)piperidine-1-carboxylate (d3) was used instead of tert-butyl 4-((2-cyano-2-(9 H -xanthen-9-yl)acetylamino)methyl)piperidine-1-carboxylate (d3) was used instead of tert-butyl 4-((2-cyano-2-(9 N -xanthen-9-yl)acetylamino)methyl)piperidine-1-carboxylate (d3) was used instead of tert-butyl 4-((2-cyano-2-(9 H -xanthen-9-yl)acetylamino)methyl)piperidine-1-carboxylate (d3) was used instead of tert-butyl 4-((2-cyano-2-(9 N -xanthen-9-yl)acetylamino)methyl)piperidine-1-carboxylate (d3) was used instead of tert-butyl 4-((2-cyano-2-(9 H -xanthen-9-yl)acetylamino)methyl)piperidine-1-carboxylate (d3) was used instead of tert-butyl 4-((2-cyano-2-(9 N -xanthen-9-yl)acetylamino)methyl)piperidine-1-carboxylate (d3) was used instead of tert-butyl 4-((2-cyano-2-(9 H -xanthen-9-yl)acetylamino)methyl)piperidine-1-carboxylate (d3) was used instead of tert-butyl 4-((2-cyano-2-(9 1 H NMR (400 MHz, Chloroform- d ) δ 8.41 (dd, J = 8.0, 1.5 Hz, 1H), 7.74 (dd, J = 8.1, 1.5 Hz, 1H),7.59-7.46 (m, 2H), 7.39-7.30 (m, 3H), 7.23 (d, J = 8.2 Hz, 2H), 7.21-7.15 (m,1H), 6.90-6.82 (m, 2H), 6.14 (t, J = 6.1 Hz, 1H), 3.75 (d, J = 6.3 Hz, 2H), 3.51(s, 2H), 3.18 (t, J = 6.3 Hz, 2H), 2.91 (d, J = 11.3 Hz, 2H), 1.99 (t, J= 11.6 Hz,2H), 1.93-1.84 (m, 2H), 1.84-1.68 (m, 4H), 1.58-1.50 (m, 2H), 1.39-1.20 (m,5H), 1.13-0.99 (m, 2H), 0.93-0.84 (m, 1H). HRMS (ESI) m / z: [M + H] + calcd forC 36 H 40 N3O3,562.3070; found, 562.3068.

[0353] Example 71

[0354] 2-Cyano- N -((4-(4-(cyclohexylmethoxy)benzyl)morpholin-2-yl)methyl)-2-(9 H -Xanthan-9-ylidene)acetamide

[0355]

[0356] The preparation method is the same as in Example 54, except that tert-butyl 4-(2-aminoethyl)piperidine-1-carboxylate in step (3) is replaced with tert-butyl 2-(aminomethyl)morpholine-4-carboxylate, and 4-(2-(2-cyano-2-(9)) in step (4) is replaced with tert-butyl 2-(aminomethyl)morpholine-4-carboxylate. H -Xanton-9-ylidene)acetamido)ethyl)piperidin-1-carboxylic acid tert-butyl ester is replaced with tert-butyl2-((2-cyano-2-(9 H -Xanton-9-ylidene)acetyl)amino)methyl)morpholine-4-carboxylate (d4), the 2-cyano- N -(2-(piperidin-4-yl)ethyl)-2-(9 H -Xanton-9-ylidene acetamide replaced with 2-cyano- N -(morpholin-2-yl)methyl)-2-(9 H -Xanton-9-ylidene)acetamide (e4), to prepare 2-cyano- N -((4-(4-(cyclohexylmethoxy)benzyl)morpholin-2-yl)methyl)-2-(9 H -Xanton-9-ylidene)acetamide, a white solid (yield 50.2%). 1 H NMR (300 MHz, Chloroform- d ) δ 8.43 (dd, J = 8.1, 1.5 Hz, 1H), 7.71 (dd, J= 8.1, 1.5 Hz, 1H),7.61-7.44 (m, 2H), 7.43-7.29 (m, 3H), 7.24-7.07 (m, 3H), 6.92-6.77 (m, 2H),6.29 (t, J = 5.6 Hz, 1H), 3.83-3.67 (m, 3H), 3.67-3.47 (m, 3H), 3.39 (s, 2H), 3.25-3.05 (m, 1H), 2.77-2.51 (m, 2H), 2.16-1.98 (m, 1H), 1.95-1.82 (m, 3H),1.81-1.63 (m, 4H), 1.34-1.21 (m, 3H), 1.14-0.99 (m, 2H). HRMS (ESI) m / z: [M +H] + calcd for C 35 H 38 N3O4, 564.2862; found, 564.2858.

[0357] Example 72

[0358] 2-Cyano- N -(2-(6-(4-(cyclohexylmethoxy)benzyl)-3,6-diazabicyclo[3.1.1]heptane-3-yl)ethyl)-2-(9 H -Xanthan-9-ylidene)acetamide

[0359]

[0360] The preparation method is the same as in Example 54, except that tert-butyl 4-(2-aminoethyl)piperidine-1-carboxylate in step (3) is replaced with tert-butyl 3-(2-aminoethyl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxylate, and 4-(2-(2-cyano-2-(9) H -Xanton-9-ylidene)acetamido)ethyl)piperidine-1-carboxylic acid tert-butyl ester is replaced with tert-butyl3-(2-(2-cyano-2-(9-) H -Xanton-9-ylidene)acetamido)ethyl)-3,6-diazabicyclo[3.1.1]heptane-6-carboxylate (d5), the 2-cyano- N -(2-(piperidin-4-yl)ethyl)-2-(9 H -Xanton-9-ylidene)acetamide replaced with N -(2-(3,6-diazabicyclo[3.1.1]heptane-3-yl)ethyl)-2-cyano-2-(9H -((4-(4-(cyclohexylmethoxy)benzyl)morpholin-2-yl)methyl)-2-(9 N -((4-(4-(cyclohexylmethoxy)benzyl)morpholin-2-yl)methyl)-2-(9 H -((4-(4-(cyclohexylmethoxy)benzyl)morpholin-2-yl)methyl)-2-(9 1 H NMR (300 MHz, Chloroform- d ) δ 8.42(d, J = 8.1 Hz, 1H), 7.74 (d, J = 8.0 Hz, 1H), 7.53 (t, J = 7.9 Hz, 1H), 7.44 (t, J = 7.7 Hz, 1H), 7.40-7.21 (m, 5H), 7.16 (t, J = 7.6 Hz, 1H), 6.83 (d, J = 8.1Hz, 2H), 3.72 (d, J = 6.1 Hz, 2H), 3.64-3.45 (m, 3H), 3.45-3.27 (m, 3H), 3.12-2.71 (m, 4H), 2.69- 2.29 (m, 3H), 1.92-1.81 (m, 2H), 1.80-1.59 (m, 5H), 1.40-1.16 (m, 3H), 1.13-0.94 (m, 2H). HRMS (ESI) m / z: [M + H] + calcd for C 37 H 41 N4O3,589.3179; found,589.3180.

[0361] Example 73

[0362] 2-((4-(4-(cyclohexylmethoxy)benzyl)morpholin-2-yl)methyl)-2-(9 H -((4-(4-(cyclohexylmethoxy)benzyl)morpholin-2-yl)methyl)-2-(9

[0363]

[0364] The preparation method refers to example 54, except that tert-butyl 3-(2- (piperidin-4-yl)ethyl)-2-(9 H - xanthen-9-yl)acetamido)ethyl)piperidine-1-carboxylate is replaced by tert-butyl 3-(2- (piperidin-4-yl)ethyl)-2-(9 H - xanthen-9-yl)acetamido)ethyl)piperidine-1-carboxylate is replaced by tert-butyl 3-(2- (piperidin-4-yl)ethyl)-2-(9 N - xanthen-9-yl)acetamido)ethyl)piperidine-1-carboxylate is replaced by tert-butyl 3-(2- (piperidin-4-yl)ethyl)-2-(9 H - xanthen-9-yl)acetamido)ethyl)piperidine-1-carboxylate is replaced by tert-butyl 3-(2- (piperidin-4-yl)ethyl)-2-(9 N - xanthen-9-yl)acetamido)ethyl)piperidine-1-carboxylate is replaced by tert-butyl 3-(2- (piperidin-4-yl)ethyl)-2-(9 H - xanthen-9-yl)acetamido)ethyl)piperidine-1-carboxylate is replaced by tert-butyl 3-(2- (piperidin-4-yl)ethyl)-2-(9 H - xanthen-9-yl)acetamido)ethyl)piperidine-1-carboxylate is replaced by tert-butyl 3-(2- (piperidin-4-yl)ethyl)-2-(9 1 H NMR (600 MHz, Chloroform- d ) δ 8.46-8.31 (m, 1H), 7.69-7.58 (m, 1H),7.53-7.45 (m, 2H), 7.43-7.34 (m, 2H), 7.35-7.28 (m, 2H), 7.28-7.23 (m, 2H),7.15 (t, J = 7.8 Hz, 1H), 6.88 (d, J = 8.2 Hz, 2H), 4.13 (s, 1H), 4.06 (s, 1H),3.98 (s, 1H), 3.94-3.87 (m, 1H), 3.81 (s, 1H), 3.72 (d, J = 6.3 Hz, 2H), 3.43(s, 1H), 3.34-3.14 (m, 2H), 2.06 (s, 1H), 1.86-1.82 (m, 2H), 1.81-1.74 (m,3H), 1.72-1.61 (m, 3H), 1.34-1.24 (m, 3H), 1.08-1.00 (m, 2H). HRMS (ESI) m / z:[M + H] + calcd for C 35 H38 N3O3, 548.2913; found,548.2914.

[0365] Example 74

[0366] 2-cyano- N -(2-(8-(4-(cyclohexylmethoxy)benzyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0367]

[0368] The preparation method refers to Example 54, except that tert-butyl 3-(2- aminoethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate is used to replace tert-butyl 4-(2-aminoethyl)piperidine-1-carboxylate in step (3), tert-butyl 3-(2-(2-cyano-2-(9 H -xanthen-9-ylidene)acetamido)ethyl)piperidine-1-carboxylate is used to replace tert-butyl 4-(2-(2-cyano-2-(9 H -xanthen-9-ylidene)acetamido)ethyl)-3,8-diazabicyclo[3.2.1]octane-8-carboxylate (d7) is used to replace 2-cyano- N -(2-(piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide in step (5), and 2-cyano- N -(2-(3,8-diazabicyclo[3.2.1]octan-3-yl)ethyl)-2-cyano-2-(9 H -xanthen-9-ylidene)acetamide (e7) is obtained, which is a white solid (the yield is 40.1%). N -(2-(8-(4-(cyclohexylmethoxy)benzyl)-3,8-diazabicyclo[3.2.1]octan-3-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide, which is a white solid (the yield is 40.1%). 1 H NMR (300MHz, Chloroform- d ) δ 8.45 (d, J = 8.1 Hz, 1H), 7.74 (d, J = 8.1 Hz, 1H), 7.55 (t, J =7.8 Hz, 1H), 7.47 (t, J= 7.8 Hz, 1H), 7.42-7.29 (m, 3H), 7.29-7.21 (m, 2H),7.16 (t, J = 7.6 Hz, 1H), 6.89-6.78 (m, 2H), 6.45 (d, J = 4.8 Hz, 1H), 3.73 (d, J = 6.2 Hz, 2H), 3.40 (s, 2H), 3.33 (d, J = 5.6 Hz, 2H), 3.01 (s, 2H), 2.39 (t, J =5.9 Hz, 2H), 2.26 (s, 4H), 1.92-1.64 (m, 9H), 1.45-1.34 (m, 2H), 1.29-1.23(m, 2H), 1.12-0.93 (m, 2H). HRMS (ESI) m / z: [M + H] + calcd for C 38 H 43 N4O3,603.3335; found,603.3339.

[0369] Example 75

[0370] 2-cyano- N -(2-(4-(4-(cyclohexylmethoxy)benzyl)-1,4-diazepan-1-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0371]

[0372] The preparation method is referred to Example 54, except that tert-butyl 4-(2- aminoethyl)piperidine-1-carboxylate in step (3) is replaced by tert-butyl 4-(2- aminoethyl)-1,4-diazepane-1-carboxylate, tert-butyl 4-(2-(2-cyano-2-(9 H -xanthen-9-ylidene)acetamido)ethyl)piperidine-1-carboxylate in step (4) is replaced by tert-butyl 4-(2-(2-cyano-2-(9 H -xanthen-9-ylidene)acetamido)ethyl)-1,4-diazepane-1-carboxylate (d8), 2-cyano- N -(2-(piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide in step (5) is replaced by N-(2-(1,4-diazacycloheptan-1-yl)ethyl)-2-cyano-2-(9 H -Xanton-9-ylidene)acetamide (e8), to prepare 2-cyano- N -(2-(4-(4-(cyclohexylmethoxy)benzyl)-1,4-diazacyclohept-1-yl)ethyl)-2-(9 H -Xanton-9-ylidene)acetamide, a white solid (yield 38.2%). 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.72 (t, J = 5.5 Hz, 1H), 8.30 (d, J = 8.0 Hz, 1H), 7.75-7.56 (m, 3H), 7.52 (d, J = 8.2 Hz, 1H), 7.45 (d, J =8.4 Hz, 2H), 7.37 (d, J = 8.1 Hz, 2H), 7.28 (t, J = 7.5 Hz, 1H), 6.98 (d, J = 8.1Hz, 2H), 4.12 (s, 2H), 3.77 (d, J = 6.0 Hz, 2H), 3.28-3.19 (m, 3H), 3.12-2.92(m, 3H), 2.89-2.79 (m, 2H), 2.77-2.68 (m, 2H), 2.66-2.56 (m, 2H), 1.84-1.73(m, 4H), 1.74-1.58 (m, 4H), 1.32-1.11 (m, 3H), 1.12-0.89 (m, 2H). HRMS (ESI)m / z: [M + H] + calcd for C 37 H 43 N4O3, 591.3335; found, 591.3339.

[0373] Example 76

[0374] 2-Cyano- N -(2-(1-(3-hydroxybenzyl)piperidin-4-yl)ethyl)-2-(9 H -Xanthan-9-ylidene)acetamide

[0375]

[0376] The preparation method refers to example 23, except that p-hydroxybenzaldehyde is replaced by 3-hydroxybenzaldehyde to prepare 2-cyano- N -(2-(1-(3-hydroxybenzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide as a white solid (yield 44.1%). 1 H NMR (300 MHz, DMSO- d 6 ) δ 9.37 (s, 1H), 8.69 (t, J = 5.5 Hz,1H), 8.29 (dd, J = 8.1, 1.5 Hz, 1H), 7.75-7.60 (m, 2H), 7.60-7.54 (m, 1H),7.54-7.48 (m, 1H), 7.48-7.38 (m, 2H), 7.32-7.21 (m, 1H), 7.12 (t, J = 7.9 Hz,1H), 6.93-6.52 (m, 3H), 3.36 (s, 2H), 3.12 (q, J = 6.6 Hz, 2H), 2.96-2.68 (m,2H), 2.13-1.68 (m, 2H), 1.68-1.47 (m, 2H), 1.37-1.23 (m, 2H), 1.22-0.97 (m,3H). HRMS (ESI) m / z: [M + H] + calcd for C 30 H 30 N3O3, 480.2287; found,480.2290.

[0377] Example 77

[0378] 2-cyano- N -(2-(1-(2-methylbenzyl)piperidin-4-yl)ethyl)-2-(9 H -xanthen-9-ylidene)acetamide

[0379]

[0380] The preparation method refers to example 1, except that benzyl bromide is replaced by 1-(bromomethyl)-2-methylbenzene to prepare 2-cyano- N-(2-(1-(2-methylbenzyl)piperidin-4-yl)ethyl)-2-(9H-xanthon-9-ylidene)acetamide, a white solid (yield 80.5%). 1 H NMR (300 MHz, DMSO- d 6 ) δ 8.68 (t, J = 5.5 Hz, 1H), 8.34-8.25(m, 1H), 7.72-7.61 (m, 2H), 7.61-7.56 (m, 1H), 7.54-7.49 (m, 1H), 7.48-7.39(m, 2H), 7.31-7.23 (m, 1H), 7.23-7.17 (m, 1H), 7.16-7.07 (m, 3H), 3.35 (s,2H), 3.12 (q, J = 6.6 Hz, 2H), 2.72 (d, J = 10.6 Hz, 2H), 2.29 (s, 3H), 1.83 (t, J = 10.4 Hz, 2H), 1.54 (d, J = 9.8 Hz, 2H), 1.27 (q, J = 6.7 Hz, 2H), 1.17-0.89 (m,3H). HRMS (ESI) m / z: [M + H] + calcd for C 31 H 32 N3O2, 478.2495; found, 478.2499.

[0381] Example 78

[0382] N -(2-(1-(4-aminobenzyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -Xanthan-9-ylidene)acetamide

[0383]

[0384] The preparation method is the same as in Example 1, except that benzyl bromide is replaced with 4-(bromomethyl)aniline to obtain... N -(2-(1-(4-aminobenzyl)piperidin-4-yl)ethyl)-2-cyano-2-(9 H -Xanton-9-ylidene)acetamide, a white solid (yield 40.7%). 1H NMR (400 MHz, DMSO- d 6 ) δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ δ 8.71 (t, J = 5.6 Hz, 1H), 8.34-8.24 (m,1H), 7.71-7.61 (m, 2H), 7.61-7.54 (m, 1H), 7.51 (d, J = 8.3 Hz, 1H), 7.47-7.38(m, 2H), 7.25 (t, J = 7.6 Hz, 1H), 7.08 (d, J = 7.9 Hz, 2H), 6.56 (d, J = 8.1 Hz,2H), 5.52-4.93 (m, 2H), 3.92-3.66 (m, 2H), 3.11 (q, J = 6.5 Hz, 2H), 3.07-2.93(m, 2H), 2.43-2.17 (m, 2H), 1.65 (d, J = 12.9 Hz, 2H), 1.41-1.24 (m, 4H),1.16-1.06 (m, 1H). HRMS (ESI) m / z: [M + H] + C 30 H 31 N4O2, 479.2442;found,479.2444。

[0385] Example 79: Study on in vitro anti-proliferative activity of compounds of the application

[0386] Cell Counting Kit-8 cell counting reagent (CCK-8) method for determining cell viability of tumor cells

[0387] The screening experiment of ZNF207 inhibitory activity of the compound is as follows: human brain glioma cells (LN229, U251MG and HS683) (purchased from the Chinese Academy of Sciences Cell Bank) are inoculated in a 96-well plate, about 100 μL per well, and 3 repeats are made for the same sample. The culture plate is placed in an incubator for a period of time (37℃, 5% CO2), different concentrations of compounds are added to each well of the culture plate, and the culture plate is placed in an incubator for a period of time. 10 μL of CCK-8 solution is added to each well, and the culture plate is placed in an incubator for 1-4 h. The absorbance (OD) at 450 nm is measured by an enzyme marker.

[0388] Cell viability (%) = [A (drug) - A (blank)] / [A (0 drug) - A (blank)] x 100.

[0389] A (drug): OD value of the well with cells, CCK-8 solution and drug solution.

[0390] A (0 drug): OD value of the well with cells, CCK-8 solution but without drug solution.

[0391] A (blank): OD value of the well without cells.

[0392] The obtained cell viability data were fitted by Graphpad Prism software to obtain the half maximal inhibitory concentration (IC 50 ).

[0393] Half maximal inhibitory concentration (IC 50 ): The smaller the value, the better the effect of the compound on anti-cell proliferation and cytotoxicity. The results are shown in Table 1.

[0394] Table 1. Anti-proliferative activity of compounds on cells (μM)

[0395]

[0396] Example 80

[0397] Verification of target binding of representative compounds to ZNF207

[0398] Kinetic determination was performed at 25 °C using a biomolecular interaction analysis system (Biacore S200). The running buffer consisted of 10 mM HEPES (pH 7.4), 150 mM NaCl, 3 mM EDTA, 0.005% (v / v) Tween 20, and supplemented with 5% (v / v) dimethyl sulfoxide (DMSO). The immobilization condition was optimized by pH screening, and 10 mM acetate buffer at pH 4.0, 4.5, 5.0 and 5.5 were tested. The ZNF207 protein was covalently immobilized on the surface of a CM5 sensor chip using standard amine coupling method, and the immobilization amount was about 3000 response units (RU). The test compound was gradient diluted and flowed through the chip surface at a flow rate of 30 μL / min, with a binding time of 60 seconds and a dissociation time of 80 seconds. The running buffer was used as a blank control, and the solvent correction method was used to eliminate the reference error. Biacore S200 Evaluation software was used for kinetic analysis to calculate the equilibrium dissociation constant (K D ). The smaller the K D value, the higher the affinity of the compound to the ZNF207 protein. The specific data are listed in Table 2.

[0399] Table 2 Kd of representative compounds to ZNF207 D Values

[0400]

[0401] The results show that the compounds of the present application have a higher affinity to ZNF207, indicating that the test compounds have the ability to bind to ZNF207.

[0402] As mentioned above, although the present application has been indicated and described with reference to particular preferred embodiments, it is by no means intended to be limited thereto. Various changes in form and details can be made thereto without departing from the spirit and scope of the application as defined in the following claims.

Claims

1. A compound having the structure shown in formula (I) or a pharmaceutically acceptable salt thereof: in: Ring A is selected from , , , , , , ; X is selected from n is any integer between 1 and 2; R 1 Selected from , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , .

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, The pharmaceutically acceptable salt is a salt formed by the compound with any of the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, carbonic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, malic acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid, mandelic acid, or ferulic acid.

3. A compound or a pharmaceutically acceptable salt thereof, characterized in that, The compound is selected from any of the following compounds: 。 4. A method for preparing the compound of claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, Includes the following steps: (1) Compound a was reacted with ethyl cyanoacetate via a Knoevenagel condensation reaction to prepare compound b; (2) Compound b was hydrolyzed with lithium hydroxide to obtain compound c; (3) Compound C and Compound d was prepared by acid-amine condensation reaction; (4) Compound d is deprotected by the Boc protecting group to obtain compound e; (5) Compound e and The reaction yields the target compound; Where A, X, n, R 1 The definition is as described in claim 1.

5. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises any one of the compounds of claims 1 to 3 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

6. The use of any compound of claims 1 to 3 or a pharmaceutically acceptable salt thereof in the preparation of an anticancer drug, wherein the cancer is breast cancer, colorectal cancer, hepatocellular carcinoma or glioma.

Images