Substituted quinoxaline and quinazolinone compounds and application thereof

Abstract

The invention discloses a substituted quinoxaline and quinazolinone compound and application thereof. The substituted quinoxaline and quinazolinone compound has the following structural formula (I) or pharmaceutically acceptable salt of the compound: # imgabs0. The compound disclosed by the invention has remarkable inhibitory activity on LSD1 and also shows remarkable anti-proliferative activity on liver cancer SMMC-7721; the compound has a novel structure, can be used as a novel and powerful LSD1 inhibitor, and lays a foundation for research and development of novel LSD1-targeted antitumor drugs.

Description

Technical Field

[0001] The present invention relates to the field of medicine, in particular to novel LSD1 inhibitor compounds, pharmaceutical compositions comprising the compounds, and substituted quinoxaline and quinazolinone compounds and their uses for reducing or inhibiting LSD1 activity in cells or subjects and preventing or treating cell proliferative disorders and / or LSD1-related disorders in subjects using the compounds and compositions. Background Art

[0002] Lysine specific demthylase 1 (LSD1), also known as KIAA0601, KDM1, AOF2 or BHC110, is a flavin adenine dinucleotide (FAD)-dependent monoamine oxidase that can specifically remove mono- and di-methyl groups from histone H3 lysine 4 / 9 (H3K4 / 9). LSD1 consists of three structural parts, including the SWIRM domain at the N-terminus, the central protruding Tower domain, and the amino oxidase like domain (AOD) domain at the C-terminus. Studies have confirmed that abnormal expression of LSD1 is closely related to tumor metastasis and proliferation, and is currently one of the important targets for targeted tumor therapy. In addition, LSD1 is also involved in the occurrence and development of a variety of diseases such as neurodegenerative diseases, cardiovascular diseases, and inflammatory responses. At present, several inhibitors have entered the clinical research stage.

[0003] LSD1 exerts different physiological functions by recruiting different proteins. When it binds to proteins such as CoREST, C-terminal-blinding protein 1 (CtBP1) and nucleosome remodeling and deacetylase (NuRD) to form a complex, LSD1 exerts a transcriptional repressive effect by catalyzing the demethylation of H3K4me2 and H3K4me1; when interacting with androgen receptor (AR) and estrogen receptor (ER), LSD1 acts as a transcriptional coactivator by demethylating H3K9me2 and H3K9me1 to promote gene transcription. In addition, the neuron-specific isoform of LSD1, LSD1n, achieves transcriptional activation of neuronal regulatory genes by catalyzing the demethylation of H4K20me2. LSD1 can also regulate the expression of non-histone substrates, including DNA methyltransferase 1 (DNMT1), tumor suppressor protein p53, myosin phosphatase targeting subunit 1 (MYPT1), signal transducers and activators of transcription 3 (STAT3), and E2F transcription factor 1 (E2F1). In summary, LSD1 participates in various cellular processes, including cell differentiation and proliferation, epithelial-mesenchymal transition (EMT), by regulating the expression of histone and non-histone genes; its dysregulation is closely related to tumor invasion and metastasis, including acute myelocytic leukemia (AML), colorectal cancer (cell resource center, CRC), small cell lung cancer (SCLC), prostatic cancer (PC), etc.

[0004] According to the different binding modes and mechanisms of action of inhibitors with LSD1, the existing LSD1 inhibitors can be divided into two types: irreversible LSD1 inhibitors and reversible LSD1 inhibitors. Among them, the irreversible LSD1 inhibitors ORY-1001, ORY-2001, INCB059872, IMG-7289, GSK2879552, TAK-418 and the reversible LSD1 inhibitors CC-90011 and SP-2577 have entered the clinical stage. In the Phase I clinical trial of GSK2879552 for the treatment of SCLC, GSK2879552 caused encephalopathy and death of patients. At present, all clinical trials on GSK2879552 have been terminated. Irreversible LSD1 inhibitors may covalently bind to non-target proteins containing FAD structures, thereby increasing the risk of unexpected toxic reactions. Reversible LSD1 inhibitors bind to LSD1 in the form of hydrogen bonds, van der Waals forces and hydrophobic forces. Relatively speaking, reversible inhibitors are more conducive to avoiding adverse reactions. Therefore, developing new, highly efficient, reversible LSD1 inhibitors is the research direction of LSD1 inhibitors. The present invention discloses a series of new LSD1 inhibitors based on quinoxaline and quinazolinone, laying a foundation for developing new drugs targeting LSD1. Summary of the invention

[0005] The present invention provides a substituted quinoxaline and quinazolinone compound and its application, which has excellent LSD1 inhibitory activity and exhibits strong anti-tumor activity, and includes a compound of formula (I) or a pharmaceutically acceptable salt, solvate, isomer, ester, acid, metabolite or prodrug thereof:

[0006] The technical solution of the present invention is as follows:

[0007] A compound having the following structural formula (I) or a pharmaceutically acceptable salt thereof:

[0008]

[0009] in:

[0010] R 1 and R 2 Independently selected from hydrogen, -OH, substituted or unsubstituted 5-10 membered heterocyclic group or -N(R), the heterocyclic group contains 1-3 heteroatoms of N, O or S, substituted or unsubstituted means that the corresponding functional group has or does not have a substituent, the substituent is C1-C6 alkyl, C1-C6 alkoxy, amino, C1-C6 alkylamino; in -N(R), each R" is independently selected from hydrogen, C1-C7 fatty primary amine, C1-C7 fatty secondary amine, C1-C7 fatty tertiary amine;

[0011] W 1 and W 2Independently selected from N, CH, CF, C-NH2, C-CH3;

[0012] R 3 It is selected from substituted or unsubstituted 5-10 membered aryl, 5-10 membered heteroaryl, C3-C6 cycloalkyl, C1-C6 alkyl, 5-10 membered heterocyclic group. Substituted or unsubstituted means that the corresponding functional group has or does not have a substituent, and the substituent is C1-C6 alkyl, C1-C6 alkoxy, halogen, halogenated C1-C6 alkyl, nitro, amino, 5-10 membered aryl.

[0013] Furthermore, R 1 and R 2 independently selected from substituted or unsubstituted 5-6 membered heterocyclic group or -N(R 4 )2, the heterocyclic group contains 1-2 N heteroatoms, and the substituent is C1-C6 alkyl, C1-C6 alkoxy, amino, C1-C6 alkylamino; -N(R 4 )2, each R 4 are independently selected from propylamine, N-methylpropylamine, N,N-dimethylpropylamine;

[0014] R 3 It is optionally substituted or unsubstituted 5-10 membered aryl, 5-10 membered heteroaryl, C3-C6 cycloalkyl, C1-C6 alkyl, 5-10 membered heterocyclyl, and the substituent is C1-C3 alkyl, C1-C3 alkoxy, halogen, halogenated C1-C3 alkyl, or amino.

[0015] Furthermore, the R 1 and R 2 Independently selected from hydrogen, methyl, methoxy, hydroxyl, substituted or unsubstituted groups:

[0016]

[0017] W 1 and W 2 Independently selected from N, CH, CF, C-CH3;

[0018] R 3 is optionally substituted or unsubstituted 5-10 membered aryl or 5-10 membered heteroaryl, and the substituent is C1-C3 alkyl, C1-C3 alkoxy, halogen, halogenated C1-C3 alkyl, or amino.

[0019] Furthermore, the R 1 and R 2 Independently selected from hydrogen, methyl, methoxy, hydroxyl, substituted or unsubstituted groups:

[0020]

[0021] W 1 and W 2 Independently selected from N, CH, CF, C-CH3;

[0022] R 3 is optionally substituted or unsubstituted as follows:

[0023]

[0024] The substituent is C1-C3 alkyl, C1-C3 alkoxy, halogen, halogenated C1-C3 alkyl, amino. Further, the structure of the compound is selected from:

[0025] (S)-4-(7-(3-aminopyrrolidin-1-yl)-3-(4-methoxyphenyl)quinoxalin-2-yl)benzonitrile (I-1)

[0026] (S)-4-(7-(3-aminopyrrolidin-1-yl)-3-(4-methylphenyl)quinoxalin-2-yl)benzonitrile (I-2)

[0027] (S)-4-(7-(3-aminopyrrolidin-1-yl)-3-(4-(trifluoromethyl)phenyl)quinoxalin-2-yl)benzonitrile (I-3) (S)-4-(7-(3-aminopyrrolidin-1-yl)-3-(4-(trifluoromethoxy)phenyl)quinoxalin-2-yl)benzonitrile (I-4) (S)-4-(7-(3-aminopyrrolidin-1-yl)-3-(4-fluorophenyl)quinoxalin-2-yl)benzonitrile (I-5)

[0028] (S)-4-(7-(3-aminopyrrolidin-1-yl)-3-(4-pyridyl)quinoxalin-2-yl)benzonitrile (I-6)

[0029] (S)-4-(7-(3-aminopiperidin-1-yl)-3-(4-methoxyphenyl)quinoxalin-2-yl)benzonitrile (I-7)

[0030] (S)-4-(7-(3-aminopiperidin-1-yl)-3-(4-methylphenyl)quinoxalin-2-yl)benzonitrile (I-8)

[0031] (S)-4-(7-(3-aminopiperidin-1-yl)-3-(4-trifluoromethylphenyl)quinoxalin-2-yl)benzonitrile (I-9) (S)-4-(7-(3-aminopiperidin-1-yl)-3-(4-trifluoromethoxyphenyl)quinoxalin-2-yl)benzonitrile (I-10)

[0032] (S)-4-(7-(3-aminopiperidin-1-yl)-3-(4-fluorophenyl)quinoxalin-2-yl)benzonitrile (I-11)

[0033] (S)-4-(7-(3-aminopiperidin-1-yl)-3-(4-pyridine)quinoxalin-2-yl)benzonitrile (I-12)

[0034] 4-(7-(3-aminopiperazin-1-yl)-3-(4-methoxyphenyl)quinoxalin-2-yl)benzonitrile (I-13)

[0035] 4-(7-(3-aminopiperazin-1-yl)-3-(4-methylphenyl)quinoxalin-2-yl)benzonitrile (I-14)

[0036] 4-(7-(3-aminopiperazin-1-yl)-3-(4-fluorophenyl)quinoxalin-2-yl)benzonitrile (I-15)

[0037] 4-(7-(3-aminopiperazin-1-yl)-3-(4-pyridyl)quinoxalin-2-yl)benzonitrile (I-16)

[0038] 4-(7-(Aminomethyl)-3-(4-methoxyphenyl)quinoxalin-2-yl)benzonitrile (I-17)

[0039] 4-(7-(Aminomethyl)-3-(4-methylphenyl)quinoxalin-2-yl)benzonitrile (I-18)

[0040] 4-(7-(Aminomethyl)-3-(4-(trifluoromethyl)phenyl)quinoxalin-2-yl)benzonitrile (I-19)

[0041] 4-(7-(Aminomethyl)-3-(4-(trifluoromethoxy)phenyl)quinoxalin-2-yl)benzonitrile (I-20)

[0042] 4-(7-(Aminomethyl)-3-(4-fluorophenyl)quinoxalin-2-yl)benzonitrile (I-21)

[0043] 4-(7-(Aminomethyl)-3-(4-pyridyl)quinoxalin-2-yl)benzonitrile (I-22)

[0044] 4-(6,7-Dihydroxy-3-(p-tolyl)quinoxalin-2-yl)benzonitrile (I-23)

[0045] (R)-4-(7-(3-aminopyrrolidin-1-yl)-3-(p-tolyl)quinoxalin-2-yl)benzonitrile (I-24)

[0046] The molecular structural formulas of the above compounds are as follows:

[0047]

[0048]

[0049] Furthermore, the pharmaceutically acceptable salt is an acid addition salt formed by the compound of general formula (I) and the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid or succinic acid, fumaric acid, salicylic acid, phenylacetic acid or mandelic acid.

[0050] The method for preparing the compound, R 2 For hydrogen, comprising the following steps:

[0051] (1) Under the action of SeO2, the reaction generates

[0052] (2) With amine intermediates Condensation

[0053] (3) The nitro group is reduced and the cyclization is carried out simultaneously to obtain

[0054] (4) and Response

[0055] (5) Hydrolysis under acidic conditions yields

[0056] Among them, R 5 Contains Boc, R 1 To R 5 The Boc in is replaced by H.

[0057] Furthermore, in step (2), the nitro group is reduced and cyclization occurs simultaneously under the action of hydrogen / palladium on carbon or iron powder / HCl or hydrazine hydrate.

[0058] W 1 and W 2 Both CH, R 2 is hydrogen, the preparation method of compounds I-1 to I-22 and I-24 of the present invention is as follows:

[0059]

[0060] In this formula, R 5 is -CH2-NH-Boc,

[0061]

[0062]

[0063] The preparation method of the I-23 compound of the present invention is as follows:

[0064]

[0065] A pharmaceutical composition comprising the compound and a pharmaceutically acceptable salt.

[0066] Use of the compound or its pharmaceutically acceptable salt or the pharmaceutical composition in preparing a drug for preventing or treating clinical conditions related to LSD1.

[0067] The disease associated with LSD1 is selected from lung cancer, melanoma, liver cancer, kidney cancer, leukemia, prostate cancer, thyroid cancer, skin cancer, pancreatic cancer, ovarian cancer, testicular cancer, breast cancer, bladder cancer, gallbladder cancer, myelodysplastic syndrome, lymphoma, esophageal cancer, gastrointestinal cancer, astrocytoma, neuroblastoma, glioma, schwannoma, mesothelioma, non-insulin-dependent diabetes mellitus, autoimmune disease or psoriasis.

[0068] The compounds of the present invention can be prepared by the above-mentioned preparation method or a similar preparation method as above, and the corresponding raw materials can be selected according to the different substituents and the different positions of the substituents.

[0069] Beneficial effects of the present invention:

[0070] The present invention develops a novel LSD1 inhibitor, which provides a new structure and idea for the development of drugs targeting LSD1, and also provides a new option for the treatment of diseases related to LSD1. The results of pharmacological tests show that the compounds of the present invention have strong inhibitory activity against LSD1, and exhibit certain inhibitory activity against other kinases, and can be used to prevent or treat clinical diseases related to LSD1, which can be: leukemia, lymphoma (non-Hodgkin's lymphoma), Hodgkin's disease (also known as Hodgkin's lymphoma) and myeloma, for example, acute lymphocytic leukemia (ALL), acute granulocytic leukemia or acute myeloid leukemia (AML), acute promyelocytic leukemia (APL), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), chronic neutrophilic leukemia (CNL), acute undifferentiated cell leukemia (AUL), running developmental large cell lymphoma (ALCL), adult T cell ALL, with blue trilineage AML with myelodysplasia (AML / TMDS), mixed trilineage leukemia (MLL), myelodysplastic syndromes (MDSs), myelodysplasia (MPD), multiple myeloma (MM) and spinal cord sarcoma, lung cancer, melanoma, liver cancer, kidney cancer, leukemia, non-small cell lung cancer, prostate cancer, thyroid cancer, skin cancer, pancreatic cancer, ovarian cancer, testicular cancer, breast cancer, bladder cancer, gallbladder cancer, myelodysplastic syndrome, lymphoma, esophageal cancer, thyroid follicular cancer, gastrointestinal cancer, tumors of the central or peripheral nervous system (such as astrocytoma, neuroblastoma, glioma or schwannoma), mesothelioma, type II or non-insulin-dependent diabetes mellitus, autoimmune diseases, psoriasis, etc. DETAILED DESCRIPTION

[0071] The specific implementation modes of the present invention are described in detail below in combination with embodiments and specific situations.

[0072] The present invention can be specifically implemented by the following examples.

[0073] N-(4-Bromo-2-nitrophenyl)acetamide (1a)

[0074] 4-Bromo-2-nitroaniline (4.00 g, 18.43 mmol) was added to a 100 ml single-necked bottle, dissolved with glacial acetic acid (30 ml), and then acetic anhydride (7.0 ml, 73.73 mmol) was added. The mixture was heated under reflux at 100°C for 5 h. The raw material disappeared after TLC detection (ethyl acetate: petroleum ether = 1:10). Under stirring, water was added to the reaction solution until solids precipitated. The solids were filtered and purified by column chromatography using ethyl acetate: petroleum ether = 1:30 as the eluent to obtain yellow solid 1a (4.41 g). The yield was 92.4%, MS [M+H] + :258.95.

[0075] (S)-tert-Butyl (1-(4-acetylamino-3-nitrophenyl)pyrrolidin-3-yl)carbamate (2a)

[0076] Add 1a (4.00 g, 15.44 mmol), tris(dibenzylideneacetone)dipalladium (0.44 g, 0.48 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (1.80 g, 3.11 mmol), Cs2CO3 (12.56 g, 38.55 mmol) and (S)-3-tert-butyloxycarbonylaminopyrrolidine (3.44 g, 18.47 mmol) to a 120 ml pressure tube. Pump nitrogen three times, add toluene (15 ml) while passing nitrogen. Heat at 115°C for 4 h, and the raw material disappears when detected by TLC (ethyl acetate: petroleum ether = 1:3). The reaction solution was cooled to room temperature and filtered. The filter cake was washed with ethyl acetate several times. The filtrate was spin-dried and purified by column chromatography with ethyl acetate: petroleum ether = 1:9 as eluent to obtain a purple-red solid 2a (4.82 g). The yield was 85.7%. MS [M+H] + :365.16.

[0077] (S)-tert-Butyl (1-(4-acetylamino-3-nitrophenyl)piperidin-3-yl)carbamate (2b)

[0078] Using N-(4-bromo-2-nitrophenyl)acetamide (4.00 g, 15.44 mmol) and (S)-3-Boc-aminopiperidine (3.71 g, 18.53 mmol) as raw materials, the preparation method was the same as 2a to obtain purple solid 2b (4.68 g) with a yield of 80.2%, MS [M+H] + :379.18. tert-Butyl 4-(4-acetylamino-3-nitrophenyl)piperazine-1-carboxylate (2c)

[0079] Using N-(4-bromo-2-nitrophenyl)acetamide (4.00 g, 15.4 mmol) and 1-Boc-piperazine (3.45 g, 18.54 mmol) as raw materials, the preparation method was the same as 2a to obtain purple solid 2c (4.79 g) with a yield of 85.3%. MS [M+H] + :365.16.

[0080] (R)-tert-Butyl (1-(4-acetylamino-3-nitrophenyl)pyrrolidin-3-yl)carbamate (2d)

[0081] Using N-(4-bromo-2-nitrophenyl)acetamide (4.00 g, 15.4 mmol) and (R)-3-tert-butyloxycarbonylaminopyrrolidine (3.44 g, 18.47 mmol) as raw materials, the preparation method was the same as 2a to obtain purple-red solid 2d (4.82 g) with a yield of 85.7%, MS [M+H] + :365.16.

[0082] (S)-tert-Butyl (1-(4-amino-3-nitrophenyl)pyrrolidin-3-yl)carbamate (3a)

[0083] 2a (4.00 g, 10.98 mmol) was added to a 150 ml single-necked bottle, dissolved in ethanol (50 ml), and then KOH (2.46 g, 43.91 mmol) was added. The mixture was heated to reflux at 90°C for 0.5 h. The raw material disappeared after TLC detection (ethyl acetate: petroleum ether = 1:2). The reaction solution was cooled to room temperature, the solvent was evaporated under reduced pressure, and then an excess of saturated NaCl solution was added, and the mixture was extracted with EA. The organic layer was collected, dried over anhydrous Na2SO4, filtered, and the solvent was evaporated from the filtrate under reduced pressure. The mixture was purified by column chromatography using ethyl acetate: petroleum ether = 1:9 as the eluent to obtain a purple solid 3a (3.01 g). The yield was 85.3%, MS [M+H] + :323.15.

[0084] (S)-tert-Butyl (1-(4-amino-3-nitrophenyl)piperidin-3-yl)carbamate (3b)

[0085] Using 2b (4.00 g, 10.58 mmol) as raw material, the preparation method was the same as 3a to obtain purple solid 3b (3.38 g) with a yield of 95.2%, MS [M+H] + :337.17.

[0086] 4-(4-Amino-3-nitrophenyl)piperazine-1-carboxylic acid tert-butyl ester (3c)

[0087] Using 2c (4.00 g, 10.98 mmol) as raw material, the preparation method was the same as 3a to obtain purple solid 3c (3.40 g) with a yield of 96.2%. MS [M+H] + :323.15.

[0088] (R)-tert-Butyl (1-(4-amino-3-nitrophenyl)pyrrolidin-3-yl)carbamate (3d)

[0089] Using 2d (4.00 g, 10.98 mmol) as raw material, the preparation method was the same as 3a to obtain purple solid 3d (3.01 g) with a yield of 85.3%, MS [M+H] + :323.15.

[0090] (4-Amino-3-nitrophenyl)methylammonium hydrochloride (4a)

[0091] 4-Amino-3-nitrobenzene (9 g, 55.17 mmol) was added to a 250 ml single-necked bottle and dissolved in THF (30 ml). BH3-THF (60 ml, 0.59 mol) was slowly added under stirring and heated at 75°C for 10 min. After the reaction solution was cooled to room temperature, 2N HCl (10 ml) was slowly added dropwise under an ice bath and the mixture was spin-dried to obtain an orange oily solid 4a with a yield of 40.1%. MS [M+H] + :204.04. The product was directly used for the next step without further purification.

[0092] Tert-butyl (4-amino-3-nitrobenzyl)carbamate (5a)

[0093] Add crude product 4a to a 250ml single-necked bottle, dissolve it with CH2Cl2 (50ml), add triethylamine (19.8ml, 0.14mol), slowly add DMAP (1.11g, 9.10mmol) and BOC2O (32.0ml, 27.31mmol) in an ice bath, and react at room temperature for 0.5h after the addition is complete. TLC detection shows that the raw material disappears (ethyl acetate: petroleum ether = 1:6). Distill off the solvent CH2Cl2 under reduced pressure, then add water, extract with DCM, dry over anhydrous Na2SO4, filter, distill off the solvent from the filtrate under reduced pressure, and purify by column chromatography to obtain yellow solid 5a (2.5g), with a yield of 56.1%, MS [M+H] + :268.11.

[0094] 2-(4-Cyanophenyl)-2-oxoacetic acid (6a)

[0095] Add p-cyanoacetophenone (3.00 g, 20.69 mmol) to a 150 ml single-necked bottle, dissolve it with pyridine (50 ml), then add SeO2 (2.70 g, 24.33 mmol), heat to reflux at 110 ° C for 1 h, then reduce the temperature to 90 ° C and continue to heat to reflux for 3 h. TLC detection shows that the raw material disappears (ethyl acetate: petroleum ether = 1:3). After the reaction solution is cooled to room temperature, it is filtered, the filter cake is washed with EA several times, and the filtrate is evaporated under reduced pressure to remove the solvent. Under ice bath, adjust the pH of the reaction solution to 9 with NaOH solution, then extract with EA, collect the water layer, adjust the pH to 3 with 2N HCl, and evaporate water under reduced pressure to obtain a yellow-white solid 6a (3.13 g), with a yield of 86.4%, MS [M+H] + :176.02.

[0096] (S)-tert-Butyl (1-(4-(2-(4-cyanophenyl)-2-oxoacetamido)-3-nitrophenyl)pyrrolidin-3-yl)carbamate (7a)

[0097] In a 250 ml single-necked bottle, add 6a (2.00 g, 11.42 mmol), add anhydrous THF (30 ml) to dissolve it, add (COCl)2 (11.6 ml, 0.14 mol) and DMF (2.6 ml, 34.26 mmol) in an ice-water bath, after the addition, warm to room temperature and react for 0.5 h. TLC detection shows that the raw material disappears (methanol: dichloromethane = 1:10). At 20 ° C, the reaction solution is dried with an explosion-proof ball filled with desiccant. Anhydrous THF (30 ml) was added for redissolution, followed by addition of a solution of 3a (2.9 g, 9.14 mmol) and pyridine (4.6 ml, 57.09 mmol) dissolved in anhydrous THF (10 ml). The reaction was allowed to react at room temperature for 10 min. The acyl chloride disappeared after TLC detection (methanol: dichloromethane = 1:10). The reaction solution was spin-dried and water was added to precipitate a solid. The solid was filtered and purified by column chromatography to obtain a light yellow solid 7a (1.56 g). The yield was 35.1%. MS [M+H] + :480.17.

[0098] (S)-tert-Butyl (1-(3-(4-cyanophenyl)-2-oxo-1,2-dihydroquinoxalin-6-yl)pyrrolidin-3-yl)carbamate (8a)

[0099] 7a (1.50 g, 3.13 mmol) was added to a 250 ml single-necked bottle, dissolved in MeOH (150 ml), and then 10% Pd / C (150 mg) was added. After the addition, H2 was introduced to exhaust the air three times, and the reaction was carried out at room temperature for 2 hours. TLC detected the disappearance of the raw material (ethyl acetate: petroleum ether = 1:1). The reaction solution was filtered, and the solvent was evaporated from the filtrate under reduced pressure. The product was purified by column chromatography using methanol: dichloromethane = 1:100 as the eluent to obtain a light yellow solid 8a (459 mg). The yield was 34.2%, MS [M+H] + :432.19.

[0100] (S)-tert-Butyl (1-(3-(4-cyanophenyl)-2-(4-methoxyphenyl)quinoxalin-6-yl)pyrrolidin-3-yl)carbamate (9a)

[0101] 8a (100 mg, 0.23 mmol), PyBroP (216 mg, 0.46 mmol), K2CO3 (96 mg, 0.69 mmol) were added to a 120 ml pressure tube, and the mixture was evacuated with N2 for 3 times. Triethylamine (97 μl, 2.08 mmol) and 1,4-dioxane (6 ml) were added under nitrogen. The mixture was heated at 100 °C for 3 h. TLC showed that 8a was completely reacted (ethyl acetate: petroleum ether = 3:1). 4-methoxyphenylboronic acid (71 mg, 0.47 mmol), PdCl2(dppf) (8 mg, 0.01 mmol) and H2O (150 μl) were then added. The mixture was heated at 100 °C for 1 h. TLC showed that the raw material disappeared (ethyl acetate: petroleum ether = 1:3). The reaction solution was cooled to room temperature, and then diluted with dichloromethane, extracted with water, and the organic layer was collected and dried over anhydrous Na2SO4, filtered, and the filtrate was evaporated under reduced pressure to remove the solvent. The solution was purified by column chromatography using ethyl acetate: petroleum ether = 1:6 as the eluent to obtain a yellow solid 9a (109 mg). The yield was 90.0%, MS [M+H] + :522.23.

[0102] (S)-tert-Butyl (1-(3-(4-cyanophenyl)-2-(4-methylphenyl)quinoxalin-6-yl)pyrrolidin-3-yl)carbamate (9b)

[0103] 8a (100 mg, 0.23 mmol), PyBroP (216 mg, 0.46 mmol), K2CO3 (96 mg, 0.69 mmol) were added to a 120 ml pressure tube, and the mixture was evacuated with N2 for 3 times. Triethylamine (97 μl, 2.08 mmol) and 1,4-dioxane (6 ml) were added under nitrogen. The mixture was heated at 100 °C for 3 h, and the reaction of 8b was complete by TLC (ethyl acetate: petroleum ether = 3:1). Subsequently, 4-methylphenylboronic acid (64 mg, 0.46 mmol), PdCl2(dppf) (8 mg, 0.01 mmol) and H2O (150 μl) were added. The mixture was heated at 100 °C for 1 h, and the reaction was complete by TLC (ethyl acetate: petroleum ether = 1:3). After the reaction solution was cooled to room temperature, dichloromethane was added to dilute it, and water was added for extraction. The organic layer was collected, dried over anhydrous Na2SO4, and filtered. The filtrate was evaporated under reduced pressure to remove the solvent, and purified by column chromatography using ethyl acetate: petroleum ether = 1:6 as the eluent to obtain a yellow solid 9b (103 mg). The yield was 88.2%, MS [M+H] + :506.24

[0104] (S)-tert-Butyl (1-(3-(4-cyanophenyl)-2-(4-(trifluoromethyl)phenyl)quinoxalin-6-yl)pyrrolidin-3-yl)carbamate (9c)

[0105] 8a (100 mg, 0.23 mmol), PyBroP (216 mg, 0.46 mmol), K2CO3 (96 mg, 0.69 mmol) were added to a 120 ml pressure tube, and the mixture was evacuated with N2 for 3 times. Triethylamine (97 μl, 2.08 mmol) and 1,4-dioxane (6 ml) were added under nitrogen. The mixture was heated at 100 °C for 3 h. TLC indicated that 8c was completely reacted (ethyl acetate: petroleum ether = 3:1). 4-trifluoromethylphenylboronic acid (88 mg, 0.463 mmol), PdCl2(dppf) (8 mg, 0.01 mmol) and H2O (150 μl) were then added. The mixture was heated at 100 °C for 1 h. TLC indicated that the reaction was complete (ethyl acetate: petroleum ether = 1:3). The reaction solution was cooled to room temperature, diluted with dichloromethane, extracted with water, and the organic layer was collected and dried over anhydrous Na2SO4, filtered, and the filtrate was evaporated under reduced pressure to remove the solvent. The solution was purified by column chromatography using ethyl acetate: petroleum ether = 1:6 as the eluent to obtain a yellow solid 9c (124.12 mg) with a yield of 95.0%, MS [M+H] + :560.21.

[0106] (S)-tert-Butyl (1-(3-(4-cyanophenyl)-2-(4-(trifluoromethoxy)phenyl)quinoxalin-6-yl)pyrrolidin-3-yl)carbamate (9d)

[0107] 8a (100 mg, 0.23 mmol), PyBroP (216 mg, 0.46 mmol), K2CO3 (96 mg, 0.69 mmol) were added to a 120 ml pressure tube, and the mixture was evacuated with N2 for 3 times. Triethylamine (97 μl, 2.08 mmol) and 1,4-dioxane (6 ml) were added under nitrogen. The mixture was heated at 100 °C for 3 h. TLC showed that the reaction of 8d was complete (ethyl acetate: petroleum ether = 3:1). Subsequently, 4-trifluoromethoxyphenylboronic acid (95 mg, 0.46 mmol), PdCl2(dppf) (8 mg, 0.01 mmol) and H2O (150 μl) were added. The mixture was heated at 100 °C for 1 h. TLC showed that the reaction was complete (ethyl acetate: petroleum ether = 1:3). The reaction solution was cooled to room temperature, and then diluted with dichloromethane, extracted with water, and the organic layer was collected and dried over anhydrous Na2SO4, filtered, and the filtrate was evaporated under reduced pressure to remove the solvent. The solution was purified by column chromatography using ethyl acetate: petroleum ether = 1:6 as the eluent to obtain a yellow solid 9d (99.73 mg), with a yield of 74.5%, MS [M+H] + :576.20.

[0108] (S)-tert-Butyl (1-(3-(4-cyanophenyl)-2-(4-fluorophenyl)quinoxalin-6-yl)pyrrolidin-3-yl)carbamate (9e)

[0109] 8a (100 mg, 0.23 mmol), PyBroP (216 mg, 0.46 mmol), K2CO3 (96 mg, 0.69 mmol) were added to a 120 ml pressure tube, and the mixture was evacuated with N2 for 3 times. Triethylamine (97 μl, 2.08 mmol) and 1,4-dioxane (6 mL) were added under nitrogen. The mixture was heated at 100 °C for 3 h, and the reaction of 8e was complete by TLC (ethyl acetate: petroleum ether = 3:1). 4-Fluorophenylboronic acid (66 mg, 0.47 mmol), PdCl2(dppf) (8 mg, 0.01 mmol) and H2O (150 μl) were then added. The mixture was heated at 100 °C for 1 h, and the reaction was complete by TLC (ethyl acetate: petroleum ether = 1:3). The reaction solution was cooled to room temperature, diluted with dichloromethane, extracted with water, and the organic layer was collected and dried over anhydrous Na2SO4, filtered, and the filtrate was evaporated under reduced pressure to remove the solvent. The solution was purified by column chromatography using ethyl acetate: petroleum ether = 1:6 as the eluent to obtain a yellow solid 9e (94.48 mg) with a yield of 80%. MS [M+H] + :510.21.

[0110] (S)-tert-Butyl (1-(3-(4-cyanophenyl)-2-(pyridin-4-yl)quinoxalin-6-yl)pyrrolidin-3-yl)carbamate (9f)

[0111] 8a (100 mg, 0.23 mmol), PyBroP (216 mg, 0.46 mmol), K2CO3 (96 mg, 0.69 mmol) were added to a 120 ml pressure tube and evacuated with N2 for 3 times. Triethylamine (97 μl, 2.08 mmol) and 1,4-dioxane (6 ml) were added under nitrogen. The reaction was heated at 100 °C for 3 h. TLC detected that 8f was completely reacted (ethyl acetate: petroleum ether = 3:1). Subsequently, 4-pyridineboronic acid (57 mg, 0.46 mmol), PdCl2 (dppf) (8 mg, 0.01 mmol) and H2O (150 μl) were added. The reaction was heated at 100 °C for 1 h. TLC detected that the reaction was complete (ethyl acetate: petroleum ether = 1:1). The reaction solution was cooled to room temperature, diluted with dichloromethane, extracted with water, and the organic layer was collected and dried over anhydrous Na2SO4, filtered, and the filtrate was evaporated under reduced pressure to remove the solvent. The solution was purified by column chromatography using ethyl acetate: petroleum ether = 1:3 as the eluent to obtain a yellow solid 9f (103 mg), with a yield of 90.2%, MS [M+H] + :493.22.

[0112] Example 1: (S)-4-(7-(3-aminopyrrolidin-1-yl)-3-(4-methoxyphenyl)quinoxalin-2-yl)benzonitrile (I-1)

[0113] 9a (100 mg, 0.19 mmol) was added to a 25 ml single-necked bottle, dissolved with anhydrous dichloromethane (4.8 ml, 0.075 mmol), trifluoroacetic acid (1.2 ml, 0.016 mmol) was added dropwise, and the reaction was carried out at room temperature for 0.5 h. The raw material disappeared after TLC detection (ethyl acetate: petroleum ether = 1:2). Under ice bath, the pH was adjusted to 8 with saturated NaHCO3, water was added, and the mixture was extracted with dichloromethane. The organic layer was collected, dried over anhydrous Na2SO4, filtered, and the solvent was evaporated from the filtrate under reduced pressure. The filtrate was purified by column chromatography with methanol: dichloromethane = 1:40 as the eluent to obtain yellow solid I-1 (42.1 mg), with a yield of 52.1%, and a melting point of 185.1°C-188.0°C. 1 H NMR (500MHz, DMSO-d6): δ7.91(d,J=9.2Hz,1H,ArH),7.83(d,J=8.3Hz,2H,ArH),7.63(d,J=8.3Hz,2H,ArH),7.39–7.31(m,3H,ArH),6.89(d,J=8.8Hz,2H, ArH),6.82(d,J=2.6Hz,1H,ArH),3.76(s,3H,-CH3),3.67–3.44(m,4H,-CH2- ×2), 3.12 (dd, J=9.9, 4.5Hz, 1H, -CH-), 1.98 (dt, J=159.7, 6.1Hz, 2H, -CH2-); 13 C NMR (126MHz, Chloroform-d): δ159.87,151.13,148.55,147.63,144.71,142.99,135.66,131.98,131.24,131.1 0,130.50,129.81,120.33,118.77,113.88,111.91,104.25,56.16,55.32,51.30,46.43,34.47; HRMS(ESI)calcd for C 26 H 23 N5O[M+H] + :422.1902.Found:422.1947.

[0114] Example 2: (S)-4-(7-(3-aminopyrrolidin-1-yl)-3-(4-methylphenyl)quinoxalin-2-yl)benzonitrile (I-2)

[0115] Using 9b (100 mg, 0.20 mmol) as raw material, the preparation method was the same as Ⅰ-1 to obtain yellow solid 1-2 (44.4 mg) with a yield of 55.4% and a melting point of 189.5℃-192.2℃. 1H NMR (500MHz, DMSO-d6): δ7.92(dd,J=9.2,1.5Hz,1H,ArH),7.86–7.79(m,2H,ArH),7.64–7.57(m,2H,ArH),7.3 8(dt,J=9.2,2.3Hz,1H,ArH),7.29(dd,J=7.9,1.5Hz,2H,ArH),7.14(d,J=7.7Hz,2H,ArH),6.83(d,J=2.5Hz,1H ArH),3.66(q,J=5.4Hz,1H,-CH-),3.58(td,J=12.8,11.6,6.2Hz,2H,-CH2-),3.47–3.43(m,1H,-CH-),3.10( dd,J=9.8,4.5Hz,1H,-CH-),2.31(s,3H,-CH3),2.14(dd,J=12.5,6.2Hz,1H,-CH-),1.83–1.77(m,1H,-CH-); 13 C NMR (126MHz, DMSO-d6): δ151.38,149.06,147.30,144.84,143.03,138.06,136.39,135.04,132.40,131.00,1 30.04,129.84,129.18,121.25,119.13,111.34,103.32,56.58,51.39,46.72,34.61,21.27; HRMS(ESI)calcd for C 26 H 23 N5[M+H] + :406.2045.Found:406.2045.

[0116] Example 3: (S)-4-(7-(3-aminopyrrolidin-1-yl)-3-(4-(trifluoromethyl)phenyl)quinoxalin-2-yl)benzonitrile (I-3)

[0117] Using 9c (100 mg, 0.16 mmol) as raw material, the preparation method was the same as Ⅰ-1 to obtain yellow solid I-3 (50 mg) with a yield of 60.4% and a melting point of 236.2℃-239.0℃. 1H NMR (500MHz, DMSO-d6): δ7.96(d,J=9.2Hz,1H,ArH),7.86(d,J=8.1Hz,2H,ArH),7.71(d,J=8.2Hz,2H,ArH),7.64(t,J=7.9Hz,4H,ArH),7.42(dd,J =9.3,2.7Hz,1H,ArH),6.86(d,J=2.6Hz,1H,ArH),3.68–3.47(m,4H,-CH2-×2),3.13(dd,J=9.9,4.4Hz,1H,-CH-),1.98(d,J=147.2Hz,2H,-CH2-); 13 C NMR (126MHz, DMSO-d6): δ151.47,149.48,145.68,144.21,143.42,143.37,135.03,132.55,131.13,130.92,130.0 4,125.74,125.47,125.44,123.58,121.63,119.07,111.66,103.18,56.53,51.35,46.75,34.51; HRMS(ESI)calcd for C 26 H 20 F3N5[M+H] + :460.1749.Found:460.1724.

[0118] Example 4: (S)-4-(7-(3-aminopyrrolidin-1-yl)-3-(4-(trifluoromethoxy)phenyl)quinoxalin-2-yl)benzonitrile (I-4)

[0119] Using 9d (90 mg, 0.16 mmol) as raw material, the preparation method was the same as Ⅰ-1 to obtain yellow solid 1-4 (52.97 mg) with a yield of 71.2% and a melting point of 127.1℃-129.4℃. 1 H NMR (500MHz, DMSO-d6): δ7.94(d,J=9.3Hz,1H,ArH),7.85(d,J=8.1Hz,2H,ArH),7.64(d,J=8.0Hz,2H,ArH),7.53(d,J=8.7Hz,2H,ArH),7.41(dd,J=9.3,2.7 Hz,1H,ArH),7.34(d,J=8.3Hz,2H,ArH),6.85(d,J=2.6Hz,1H,ArH),3.69–3.4 6(m,4H,-CH2-×2),3.13(dd,J=10.0,4.5Hz,1H-CH-),2.14–1.83(m,2H-CH2-);13 CNMR (126MHz, DMSO-d6): δ151.39,149.31,148.62,145.92,144.36,143.31,138.54,134.99,132.48,132.09,1 31.10,129.94,121.51,121.01,119.49,119.09,111.57,103.25,56.45,51.34,46.73,34.44; HR-MS(ESI)calcd for C 26 H 20 F3N5O[M+H] + :476.1698.Found:476.1649.

[0120] Example 5: (S)-4-(7-(3-aminopyrrolidin-1-yl)-3-(4-fluorophenyl)quinoxalin-2-yl)benzonitrile (I-5)

[0121] Using 9e (90 mg, 0.18 mmol) as raw material, the preparation method was the same as Ⅰ-1 to obtain yellow solid 1-5 (36.30 mg) with a yield of 50.2% and a melting point of 92.1℃-95.2℃. 1 H NMR (500MHz, DMSO-d6): δ7.93(d,J=9.2Hz,1H,ArH),7.84(d,J=8.0Hz,2H,ArH),7.62(d,J=8.0Hz,2H, ArH),7.42(ddd,J=21.2,8.9,4.2Hz,3H,ArH),7.18(t,J=8.7Hz,2H,ArH),6.84(d,J=2.6Hz,1H,ArH), 3.67(p,J=5.4Hz,1H,-CH-),3.59(td,J=12.4,11.4,6.3Hz,2H,-CH2-),3.48–3.43(m,1H,-CH-),3.11 (dd,J=9.9,4.5Hz,1H,-CH-),2.14(dd,J=12.5,6.2Hz,1H,-CH-),1.80(dt,J=12.5,6.3Hz,1H,-CH-); 13C NMR (126MHz, DMSO-d6): δ163.44,161.49,151.37,149.19,146.35,144.53,143.20,135.74,135.71,134.97,132.45,132.3 1,132.24,131.07,129.87,121.38,119.11,115.63,115.46,111.45,103.27,56.51,51.36,46.73,34.53; HRMS(ESI)calcd forC 25 H 20 FN5[M+H] + :410.1702.Found:410.1749.

[0122] Example 6: (S)-4-(7-(3-aminopyrrolidin-1-yl)-3-(4-pyridine)quinoxalin-2-yl)benzonitrile (I-6)

[0123] Using 9f (100 mg, 0.20 mmol) as raw material, the preparation method is the same as Ⅰ-1 to obtain yellow solid I-6 (36 mg) with a yield of 45.2% and a melting point of 160.0℃-162.4℃. 1 H NMR (500MHz, DMSO-d6): δ8.55(d,J=5.0Hz,2H,ArH), 7.98(d,J=9.2Hz,1H,ArH), 7.87(d,J=7.9Hz,2H,ArH), 7.66(d,J=7.9Hz,2H,ArH), 7.39(d,J=5.1Hz,3H,ArH),6.88(d,J=2.6Hz,1H,ArH),3.72–3.49(m,4H,-CH2-×2),3.21–3.16(m,1H,-CH-),2.18–1.87(m,2H,-CH2-); 13 C NMR (126MHz, DMSO-d6): δ151.54,149.95,149.58,146.67,144.74,143.97,143.54,135.06,132.57,1 31.12,130.16,124.59,121.75,119.06,111.78,103.28,56.01,51.19,46.69,33.95; HRMS(ESI)calcd for C 24 H 20 N6[M+H] + :393.1828.Found:393.1837.

[0124] (S)-tert-Butyl (1-(4-(2-(4-cyanophenyl)-2-oxoacetamido)-3-nitrophenyl)piperidin-3-yl)carbamate (10a)

[0125] Using 6a (2 g, 11.42 mmol) and 3b (3.07 g, 9.14 mmol) as raw materials, the preparation method was the same as 7a to obtain yellow solid 10a (2.04 g) with a yield of 45.2%. MS [M+H] + :494.19.

[0126] (S)-tert-Butyl (1-(3-(4-cyanophenyl)-2-oxo-1,2-dihydroquinoxalin-6-yl)piperidin-3-yl)carbamate (11a)

[0127] Using 10a (2 g, 4.05 mmol) as the starting material, the preparation method was the same as 8a to obtain yellow solid 11a (634 mg) with a yield of 35.2%. MS [M+H] + :446.20.

[0128] (S)-tert-Butyl (1-(3-(4-cyanophenyl)-2-(4-methoxyphenyl)quinoxalin-6-yl)piperidin-3-yl)carbamate (12a)

[0129] Using 11a (100 mg, 0.22 mmol) and 4-methoxyphenylboronic acid (68 mg, 0.45 mmol) as raw materials, the preparation method was the same as 9a to obtain yellow solid 12a (102 mg) with a yield of 85.2%, MS [M+H] + :536.25.

[0130] (S)-tert-Butyl (1-(3-(4-cyanophenyl)-2-(4-methylphenyl)quinoxalin-6-yl)piperidin-3-yl)carbamate (12b)

[0131] Using 11a (100 mg, 0.22 mmol) and 4-methylphenylboronic acid (61 mg, 0.45 mmol) as raw materials, the preparation method was the same as 9a to obtain yellow solid 12b (100 mg) with a yield of 85.6%. MS [M+H] + :520.25.

[0132] (S)-tert-Butyl (1-(3-(4-cyanophenyl)-2-(4-(trifluoromethyl)phenyl)quinoxalin-6-yl)piperidin-3-yl)carbamate (12c)

[0133] Using 11a (100 mg, 0.22 mmol) and 4-trifluoromethylphenylboronic acid (85 mg, 0.45 mmol) as raw materials, the preparation method was the same as 9a to obtain yellow solid 12c (113 mg) with a yield of 87.6%. MS [M+H] + :574.23.

[0134] (S)-tert-Butyl (1-(3-(4-cyanophenyl)-2-(4-(trifluoromethoxy)phenyl)quinoxalin-6-yl)piperidin-3-yl)carbamate (12d)

[0135] Using 11a (100 mg, 0.22 mmol) and 4-trifluoromethoxyphenylboronic acid (93 mg, 0.45 mmol) as raw materials, the preparation method was the same as 9a to obtain yellow solid 12d (119 mg) with a yield of 90.2%. MS [M+H] + :590.22.

[0136] (S)-tert-Butyl (1-(3-(4-cyanophenyl)-2-(4-fluorophenyl)quinoxalin-6-yl)piperidin-3-yl)carbamate (12e)

[0137] Using 11a (100 mg, 0.22 mmol) and 4-fluorophenylboronic acid (63 mg, 0.45 mmol) as raw materials, the preparation method was the same as 9a to obtain yellow solid 12e (100 mg) with a yield of 85.4%. MS [M+H] + :524.23.

[0138] (S)-tert-Butyl (1-(3-(4-cyanophenyl)-2-(pyridin-4-yl)quinoxalin-6-yl)piperidin-3-yl)carbamate (12f)

[0139] Using 11a (100 mg, 0.22 mmol) and pyridine-4-boronic acid (55 mg, 0.45 mmol) as raw materials, the preparation method was the same as 9a to obtain yellow solid 12f (101 mg), yield 89.2% MS [M+H] + :507.23.

[0140] Example 7: (S)-4-(7-(3-aminopiperidin-1-yl)-3-(4-methoxyphenyl)quinoxalin-2-yl)benzonitrile (I-7)

[0141] Using 12a (100 mg, 0.19 mmol) as raw material, the preparation method is the same as I-1 to obtain yellow solid I-7 (48.93 mg) with a yield of 60.2% and a melting point of 132.6°C-135.4°C. 1H NMR (500MHz, DMSO-d6): δ7.92(d,J=9.3Hz,1H,ArH),7.85(d,J=8.0Hz,2H,ArH),7.73(dd,J=9.4,2.8Hz,1 H,ArH),7.63(d,J=8.0Hz,2H,ArH),7.34(d,J=8.4Hz,2H,ArH),7.26(d,J=2.7Hz,1H,ArH),6.91(d,J=8.4 Hz,2H,ArH),3.95–3.84(m,2H,-CH2-),3.77(s,3H,-OCH3),2.96–2.82(m,2H,-CH2-),2.71(dd,J=12.5,9 .5Hz,1H,-CH-),1.89(s,2H,-CH2),1.78–1.54(m,2H,-CH2-),1.27(ddd,J=23.8,10.9,5.0Hz,2H,-NH2-); 13 C NMR (126MHz, DMSO-d6): δ159.93,151.84,151.49,148.89,144.72,142.62,136.10,132.48,131.57,131.20,130. 97,129.60,123.64,119.13,114.10,111.45,108.18,55.64,54.67,48.32,47.11,31.57,23.15; HRMS(ESI)calcd for C 27 H 25 N5O[M+H] + :436.2137.Found:437.2149.

[0142] Example 8: (S)-4-(7-(3-aminopiperidin-1-yl)-3-(4-methylphenyl)quinoxalin-2-yl)benzonitrile (I-8)

[0143] Using 12b (100 mg, 0.19 mmol) as raw material, the preparation method is the same as I-1 to obtain yellow solid I-8 (53.53 mg) with a yield of 66.3% and a melting point of 209.5°C-210.2°C. 1H NMR (500MHz, DMSO-d6): δ7.93 (d, J=9.3Hz, 1H, ArH), 7.84 (d, J=8.1Hz, 2H, ArH), 7.74 (dd, J=9. 4,2.8Hz,1H,ArH),7.63(d,J=8.1Hz,2H,ArH),7.33–7.25(m,3H,ArH),7.16(d,J=7.9Hz,2H,Ar H),3.99–3.86(m,2H,-CH2-),2.95–2.79(m,2H,-CH2-),2.69(dd,J=12.5,9.6Hz,1H,-CH-),2. 32(s,3H,-CH3),1.98–1.82(m,2H,-CH2-),1.79–1.54(m,2H,-CH2-),1.40–1.11(m,2H,-NH2); 13 C NMR (126MHz, DMSO-d6): δ152.03,151.49,148.85,144.65,142.87,138.32,136.21,135.91,132.43,131.00,130. 07,129.60,129.21,123.50,119.10,111.42,107.57,56.69,48.24,47.55,33.66,23.66,21.28; HRMS(ESI)calcd for C 27 H 25 N5[M+H] + :420.2188.Found:421.2158.

[0144] Example 9: (S)-4-(7-(3-aminopiperidin-1-yl)-3-(4-trifluoromethylphenyl)quinoxalin-2-yl)benzonitrile (I-9)

[0145] Using 12c (100 mg, 0.17 mmol) as raw material, the preparation method is the same as I-1 to obtain yellow solid I-9 (46 mg) with a yield of 55.4% and a melting point of 200.2℃-202.6℃. 1H NMR (500MHz, DMSO-d6): δ7.96 (d, J=9.4Hz, 1H, ArH), 7.87 (d, J=8.1Hz, 2H, ArH), 7.78 (dd, J=9.5, 2.8Hz,1H,ArH),7.73(d,J=8.2Hz,2H,ArH),7.64(dd,J=8.3,3.6Hz,4H,ArH),7.30(d,J=2.7Hz,1H ,ArH),4.00–3.91(m,2H,-CH2-),2.97–2.79(m,2H,CH2-),2.71(dd,J=12.4,9.6Hz,1H,-CH-),1.9 0(d,J=13.5Hz,2H,-CH2-),1.66(ddd,J=101.9,9.7,5.0Hz,2H,-CH2-),1.34–1.21(m,2H,-NH2-); 13 C NMR (126MHz, DMSO-d6): δ152.36,151.57,147.23,144.04,143.33,143.23,135.80,132.57,131.14,130.97,129.81,1 25.72,125.52,125.48,123.62,123.55,119.06,111.74,107.17,56.55,48.14,47.57,33.70,23.66; HRMS(ESI)calcd for C 27 H 22 F3N5[M+H] + :474.1906.Found:474.1925.

[0146] Example 10: (S)-4-(7-(3-aminopiperidin-1-yl)-3-(4-trifluoromethoxyphenyl)quinoxalin-2-yl)benzonitrile (I-10)

[0147] Using 12d (100 mg, 0.17 mmol) as raw material, the preparation method is the same as I-1 to obtain yellow solid I-10 (52 mg) with a yield of 62.4% and a melting point of 149.2°C-152.9°C. 1H NMR (500MHz, DMSO-d6): δ7.95 (d, J=9.4Hz, 1H, ArH), 7.86 (d, J=8.1Hz, 2H, ArH), 7.77 (dd, J=9. 4,2.8Hz,1H,ArH),7.64(d,J=8.1Hz,2H,ArH),7.54(d,J=8.6Hz,2H,ArH),7.36(d,J=8.2Hz,2H, ArH),7.29(d,J=2.7Hz,1H,ArH),3.99–3.89(m,2H,-CH2-),2.89(ddd,J=51.0,9.5,3.3Hz,2H,- CH2-),2.72(dd,J=12.5,9.6Hz,1H,-CH-),1.92–1.75(m,2H,-CH2-),1.58–1.29(m,2H,-CH2-); 13 C NMR (126MHz, DMSO-d6): δ152.18,151.55,147.64,144.14,143.11,138.33,135.90,132.51,132.16,131.10,129. 75,123.69,121.52,121.05,119.48,119.07,111.67,107.68,55.21,48.21,47.26,32.28,23.31; HRMS(ESI)calcd for C 27 H 22 F3N5O[M+H] + :490.1885.Found:490.1837.

[0148] Example 11: (S)-4-(7-(3-aminopiperidin-1-yl)-3-(4-fluorophenyl)quinoxalin-2-yl)benzonitrile (I-11)

[0149] Using 12e (100 mg, 0.19 mmol) as raw material, the preparation method is the same as I-1 to obtain yellow solid I-11 (75 mg) with a yield of 93.2% and a melting point of 187.7°C-190.2°C. 1H NMR (500MHz, DMSO-d6): δ7.95 (d, J=9.4Hz, 1H, ArH), 7.85 (d, J=8.1Hz, 2H, ArH), 7.76 (dd,J=9.4,2.8Hz,1H,ArH),7.63(d,J=8.1Hz,2H,ArH),7.45(dd,J=8.6,5.6Hz,2H,A rH),7.29(d,J=2.7Hz,1H,ArH),7.20(t,J=8.9Hz,2H,ArH),3.96–3.88(m,2H,-CH2-) ,2.99–2.87(m,2H,-CH2-),2.79(dd,J=12.5,9.5Hz,1H,-CH-),1.89(s,4H,-CH2-×2); 13 C NMR (126MHz, DMSO-d6): δ163.56,161.60,152.14,151.50,147.89,144.36,143.07,135.81,135.58,135.55,132.48,132.37,1 32.30,131.07,129.64,123.55,119.09,115.69,115.51,111.53,107.45,56.70,48.21,47.58,33.73,23.66; HRMS(ESI)calcd for C 26 H 22 FN5[M+H] + :424.1937.Found:424.1928.

[0150] Example 12: (S)-4-(7-(3-aminopiperidin-1-yl)-3-(4-pyridine)quinoxalin-2-yl)benzonitrile (I-12)

[0151] Using 12f (100 mg, 0.20 mmol) as raw material, the preparation method is the same as I-1 to obtain yellow solid I-12 (51.70 mg) with a yield of 64.5% and a melting point of 162.4°C-165.4°C. 1H NMR (500MHz, DMSO-d6): δ8.58–8.53(m,2H,ArH),7.98(d,J=9.4Hz,1H,ArH),7.88(d,J =8.1Hz,2H,ArH),7.80(dd,J=9.5,2.8Hz,1H,ArH),7.66(d,J=8.1Hz,2H,ArH),7.42–7. 37(m,2H,ArH),7.30(d,J=2.7Hz,1H,ArH),4.04–3.93(m,2H,-CH2-),3.00–2.94(m,1H ,-CH-),2.84–2.71(m,2H,-CH2-),2.01–1.88(m,2H,-CH2-),1.80–1.55(m,2H,-CH2-); 13 C NMR (126MHz, DMSO-d6): δ152.47,151.56,149.99,146.55,146.17,143.80,143.50,135.75,132.57,131.11, 129.89,124.60,123.61,119.04,111.84,107.01,60.22,56.56,48.09,47.61,33.77,23.68; HRMS(ESI)calcd for C 25 H 22 N6[M+H] + :407.1905.Found:407.1959.

[0152] (S)-tert-Butyl (1-(4-(2-(4-cyanophenyl)-2-oxoacetamido)-3-nitrophenyl)piperazin-3-yl)carbamate (13a)

[0153] Using 6a (2 g, 11.42 mmol) and 3c (2.94 g, 9.13 mmol) as raw materials, the preparation method was the same as 7a to obtain yellow solid 13a (2.08 g) with a yield of 48.4%, MS [M+H] + :480.17.

[0154] Tert-butyl (1-(3-(4-cyanophenyl)-2-oxo-1,2-dihydroquinoxalin-6-yl)piperazin-3-yl)carbamate (14a)

[0155] Using 13a (2 g, 4.17 mmol) as starting material, the preparation method was the same as 8a to obtain yellow solid 14a (633 mg) with a yield of 35.2%. MS [M+H] + :432.19.

[0156] Tert-butyl (1-(3-(4-cyanophenyl)-2-(4-methoxyphenyl)quinoxalin-6-yl)piperazin-3-yl)carbamate (15a)

[0157] Using 14a (100 mg, 0.23 mmol) and 4-methoxyphenylboronic acid (70 mg, 0.46 mmol) as raw materials, the preparation method was the same as 9a to obtain yellow solid 15a (102 mg) with a yield of 84.2%, MS [M+H] + :522.23.

[0158] Tert-butyl (1-(3-(4-cyanophenyl)-2-(4-methylphenyl)quinoxalin-6-yl)piperazin-3-yl)carbamate (15b)

[0159] Using 14a (100 mg, 0.23 mmol) and 4-methylphenylboronic acid (63 mg, 0.46 mmol) as raw materials, the preparation method was the same as 9a to obtain yellow solid 15b (93 mg) with a yield of 79.3%. MS [M+H] + :506.24.

[0160] Tert-butyl (1-(3-(4-cyanophenyl)-2-(4-fluorophenyl)quinoxalin-6-yl)piperazin-3-yl)carbamate (15c)

[0161] Using 14a (100 mg, 0.23 mmol) and 4-fluorophenylboronic acid (65 mg, 0.46 mmol) as raw materials, the preparation method was the same as 9a to obtain yellow solid 15c (86 mg) with a yield of 72.5%, MS [M+H] + :510.21.

[0162] Tert-butyl (1-(3-(4-cyanophenyl)-2-(pyridin-4-yl)quinoxalin-6-yl)piperazin-3-yl)carbamate (15d)

[0163] Using 14a (100 mg, 0.23 mmol) and pyridine-4-boronic acid (57 mg, 0.46 mmol) as raw materials, the preparation method was the same as 9a to obtain yellow solid 15d (97 mg) with a yield of 85.2%, MS [M+H] + :493.22.

[0164] Example 13: 4-(7-(3-aminopiperazin-1-yl)-3-(4-methoxyphenyl)quinoxalin-2-yl)benzonitrile (I-13)

[0165] Using 15a (100 mg, 0.19 mmol) as raw material, the preparation method is the same as I-1 to obtain yellow solid I-13 (50 mg) with a yield of 62.3% and a melting point of 155.5°C-158.3°C.1 H NMR (500MHz, DMSO-d6): δ7.95(d,J=9.3Hz,1H,ArH),7.85(d,J=8.1Hz,2H,ArH),7.77(dd,J=9.3,2.8Hz,1H,ArH),7.64(d,J=8.2Hz,2H,ArH),7.35( d,J=8.7Hz,2H,ArH),7.29(d,J=2.7Hz,1H,ArH),6.91(d,J=8.7Hz,2H,ArH),3.77(s,3H,-OCH3),2.97(t,J=5.1Hz,4H,-CH2-×2),1.23(s,1H,-NH-); 13 C NMR (126MHz, DMSO-d6): δ159.96,152.44,151.51,149.12,144.70,142.45,136.33,132.48,131.60,13 1.17,130.98,129.48,123.47,119.13,114.10,111.46,108.35,55.64,48.29,45.21; HRMS(ESI)calcd for C 26 H 23 N5O[M+H] + :422.1981.Found:422.1946.

[0166] Example 14: 4-(7-(3-aminopiperazin-1-yl)-3-(4-methylphenyl)quinoxalin-2-yl)benzonitrile (I-14)

[0167] Using 15b (100 mg, 0.20 mmol) as raw material, the preparation method is the same as I-1 to obtain yellow solid I-14 (46.85 mg) with a yield of 58.4% and a melting point of 230.1°C-232.4°C. 1 H NMR (500MHz, DMSO-d6): δ7.96(d,J=9.3Hz,1H,ArH),7.84(d,J=8.0Hz,2H,ArH),7.78(dd,J=9.4,2.8Hz,1H,ArH),7.63(d,J=8.0Hz,2H,ArH),7 .34–7.27(m,3H,ArH),7.16(d,J=7.9Hz,2H,ArH),3.39(s,4H,-CH2-×2),2.95(t,J=3.8Hz,4H,-CH2-×2),2.32(s,3H,-CH3),1.23(s,1H,-NH-); 13C NMR (126MHz, DMSO-d6): δ152.53,151.60,149.38,144.57,142.59,138.44,136.31,136.14,132.45,131 .01,130.11,129.55,129.23,123.48,119.09,111.48,108.28,48.22,45.18,21.29; HRMS(ESI)calcdfor C 26 H 23 N5[M+H] + :406.2152.Found:406.2110.

[0168] Example 15: 4-(7-(3-aminopiperazin-1-yl)-3-(4-fluorophenyl)quinoxalin-2-yl)benzonitrile (I-15)

[0169] Using 15c (100 mg, 0.20 mmol) as raw material, the preparation method is the same as I-1 to obtain yellow solid I-15 (64 mg) with a yield of 80.1% and a melting point of 121.9°C-124.5°C. 1 H NMR (500MHz, DMSO-d6): δ7.97(d,J=9.4Hz,1H,ArH),7.86(d,J=8.1Hz,2H,ArH),7.80(dd,J=9.4,2.8Hz,1H,ArH),7.64(d,J=8.0Hz,2H,ArH),7 .46(dd,J=8.6,5.5Hz,2H,ArH),7.29(d,J=2.7Hz,1H,ArH),7.21(t,J=8.7Hz,2H,ArH),3.39(s,4H,-CH2-×2),2.93(t,J=5.0Hz,4H,-CH2-×2); 13 C NMR (126MHz, DMSO-d6) δ161.64,152.83,151.55,148.33,144.29,142.81,136.16,135.51,132.49,132.41 ,132.34,131.08,129.54,123.53,119.08,115.70,115.53,111.57,108.01,48.64,45.53; HRMS(ESI)calcd for C 25 H 20 FN5[M+H] + :410.1702.Found:410.1705.

[0170] Example 16: 4-(7-(3-aminopiperazin-1-yl)-3-(4-pyridine)quinoxalin-2-yl)benzonitrile (I-16)

[0171] Using 15d (100 mg, 0.20 mmol) as raw material, the preparation method is the same as I-1 to obtain yellow solid I-16 (48 mg) with a yield of 60.7% and a melting point of 131.7°C-132.2°C. 1 H NMR (500MHz, Methanol-d4): δ8.53–8.49(m,2H,-CH=),8.01(d,J=9.5Hz,1H,-CH=),7.79(dd,J=9.4,2.8Hz,1H,-CH=),7.76–7.73(m,2H,Ar H),7.66(d,J=8.3Hz,2H,ArH),7.50–7.47(m,2H,ArH),7.34(d,J=2.7Hz,1H,ArH),3.50–3.46(m,4H,-CH2-×2),3.08–3.03(m,4H,-CH2-×2); 13 C NMR (126MHz, DMSO-d6): δ153.21,151.59,150.00,146.65,146.50,143.72,143.24,136.09,13 2.58,131.13,129.76,124.62,123.61,119.03,111.88,107.61,48.58,45.59; HRMS(ESI)calcd for C 24 H 20 N6[M+H] + :393.1828.Found:393.1802.

[0172] Tert-butyl (4-(2-(4-cyanophenyl)-2-oxoacetylamino)-3-nitrobenzyl)carbamate (16a)

[0173] Using 6a (2 g, 11.42 mmol) and 5a (2.44 g, 9.14 mmol) as raw materials, the preparation method was the same as 7a to obtain yellow solid 16a (1.41 g) with a yield of 36.4%, MS [M+H] + :425.13.

[0174] Tert-butyl (3-(4-cyanophenyl)-2-oxo-1,2-dihydroquinoxalin-6-yl)methyl)carbamate (17a)

[0175] Using 16a (1.4 g, 3.30 mmol) as starting material, the preparation method was the same as 8a to obtain yellow solid 17a (437 mg) with a yield of 35.2%, MS [M+H] + :377.14.

[0176] Tert-butyl (3-(4-cyanophenyl)-2-(4-methoxyphenyl)quinoxalin-6-yl)methyl)carbamate (18a)

[0177] Using 17a (100 mg, 0.27 mmol) and 4-methoxyphenylboronic acid (81 mg, 0.53 mmol) as raw materials, the preparation method was the same as 9a to obtain yellow solid 18a (111 mg) with a yield of 89.5%, MS [M+H] + :467.19.

[0178] Tert-butyl (3-(4-cyanophenyl)-2-(4-methylphenyl)quinoxalin-6-yl)methyl)carbamate (18b)

[0179] Using 17a (100 mg, 0.27 mmol) and 4-methylphenylboronic acid (72 mg, 0.53 mmol) as raw materials, the preparation method was the same as 9a to obtain yellow solid 18b (103 mg) with a yield of 86.4%. MS [M+H] + :451.20.

[0180] Tert-butyl (3-(4-cyanophenyl)-2-(4-(trifluoromethyl)phenyl)quinoxalin-6-yl)methyl)carbamate (18c)

[0181] Using 17a (100 mg, 0.27 mmol) and 4-trifluoromethylphenylboronic acid (101 mg, 0.53 mmol) as raw materials, the preparation method was the same as 9a to obtain yellow solid 18c (115 mg) with a yield of 85.7%, MS [M+H] + :505.17.

[0182] Tert-butyl (3-(4-cyanophenyl)-2-(4-(trifluoromethoxy)phenyl)quinoxalin-6-yl)methyl)carbamate (18d)

[0183] Using 17a (100 mg, 0.267 mmol) and 4-trifluoromethoxyphenylboronic acid (109 mg, 0.53 mmol) as raw materials, the preparation method was the same as 9a to obtain yellow solid 18d (131 mg) with a yield of 94.8%, MS [M+H] + :521.16.

[0184] Tert-butyl (3-(4-cyanophenyl)-2-(4-fluorophenyl)quinoxalin-6-yl)methyl)carbamate (18e)

[0185] Using 17a (100 mg, 0.27 mmol) and 4-fluorophenylboronic acid (74 mg, 0.53 mmol) as raw materials, the preparation method was the same as 9a to obtain yellow solid 18e (103 mg) with a yield of 85.2%. MS [M+H] + :455.17.

[0186] Tert-butyl (3-(4-cyanophenyl)-2-(4-pyridyl)quinoxalin-6-yl)methyl)carbamate (18f)

[0187] Using 17a (100 mg, 0.27 mmol) and pyridine-4-boronic acid (65 mg, 0.53 mmol) as raw materials, the preparation method was the same as 9a to obtain yellow solid 18f (100 mg) with a yield of 86.1%. MS [M+H] + :438.18.

[0188] Example 17: 4-(7-(Aminomethyl)-3-(4-methoxyphenyl)quinoxalin-2-yl)benzonitrile (I-17)

[0189] Using 18a (100 mg, 0.21 mmol) as raw material, the preparation method is the same as I-1 to obtain yellow solid I-17 (58 mg) with a yield of 73.4% and a melting point of 76.3°C-78.9°C. 1 H NMR (500MHz, DMSO-d6): δ8.12–8.04(m,2H,ArH),7.88(d,J=8.3Hz,3H,ArH),7.68(d,J=8.3Hz,2H,ArH),7 .42(d,J=8.7Hz,2H,ArH),6.94(d,J=8.7Hz,2H,ArH),4.00(s,2H,-CH2-),3.78(s,3H,-OCH3),1.23(s,2H 2H,-NH2); 13 C NMR (126MHz, DMSO-d6): δ160.34,152.25,151.68,146.36,144.36,140.62,140.37,132.55,131.82 ,131.60,131.02,130.83,128.73,126.23,119.08,114.17,111.68,55.68,45.49; HRMS(ESI)calcd for C 23 H 18 N4O[M+H] + :367.1180.Found:367.1148.

[0190] Example 18: 4-(7-(Aminomethyl)-3-(4-methylphenyl)quinoxalin-2-yl)benzonitrile (I-18)

[0191] Using 18b (100 mg, 0.22 mmol) as raw material, the preparation method is the same as I-1 to obtain yellow solid I-18 (51 mg) with a yield of 65.4% and a melting point of 165.1°C-166.5°C. 1 H NMR (500MHz, DMSO-d6): δ8.11–8.07(m,2H,ArH),7.90–7.85(m,3H,ArH),7.67(d,J=8.2Hz,2H,ArH),7.37(d,J=7 .9Hz,2H,ArH),7.19(d,J=7.9Hz,2H,ArH),4.00(s,2H,-CH2-),2.33(s,3H,-CH3),2.18(d,J=26.9Hz,2H,-NH2); 13 CNMR (126MHz, DMSO-d6): δ152.53,151.72,144.23,140.78,140.33,139.01,135.86,132.51,13 1.63,131.06,130.24,129.29,128.76,126.04,119.05,111.69,45.71,21.32; HRMS(ESI)calcd for C 23 H 18 N4[M+H] + :351.1610.Found:351.1621.

[0192] Example 19: 4-(7-(Aminomethyl)-3-(4-(trifluoromethyl)phenyl)quinoxalin-2-yl)benzonitrile (I-19)

[0193] Using 18c (100 mg, 0.20 mmol) as raw material, the preparation method is the same as I-1 to obtain yellow solid I-19 (42 mg) with a yield of 52.1% and a melting point of 164.2°C-165.4°C. 1 H NMR (500MHz, DMSO-d6): δ8.15–8.12(m,2H,ArH),7.93(dd,J=8.7,1.9Hz,1H,ArH),7.88(d,J=8.3Hz,2 H,ArH),7.77(d,J=8.2Hz,2H,ArH),7.70(t,J=8.4Hz,4H,ArH),4.02(s,2H,-CH2),2.19(s,2H,-NH2); 13C NMR(126MHz,DMSO-d6)δ151.71,151.25,148.24,143.57,142.87,141.16,140.20,132.62,131 .90,131.23,131.18,128.87,125.97,125.59,125.56,119.00,111.98,45.81; HRMS(ESI)calcd for C 23 H 15 F3N4[M+H] + :405.1327.Found:405.1362.

[0194] Example 20: 4-(7-(Aminomethyl)-3-(4-(trifluoromethoxy)phenyl)quinoxalin-2-yl)benzonitrile (I-20)

[0195] Using 18d (100 mg, 0.19 mmol) as raw material, the preparation method is the same as I-1 to obtain yellow solid I-20 (52.83 mg) with a yield of 65.4% and a melting point of 160.1℃-163.4℃. 1 H NMR (500MHz, DMSO-d6): δ8.12(d,J=8.8Hz,2H,ArH),7.92(dd,J=8.6,1.9Hz,1H,ArH),7.90–7.87(m,2H,ArH),7.70–7 .67(m,2H,ArH),7.62–7.59(m,2H,ArH),7.40(d,J=8.3Hz,2H,ArH),4.02(s,2H,-CH2-),2.37(d,J=2.3Hz,2H,-NH2); 13 C NMR (126MHz, DMSO-d6): δ151.71,151.36,149.16,147.69,143.75,141.04,140.21,137.99,132.56,132 .39,131.82,131.19,128.82,126.05,121.51,121.09,119.47,119.02,111.89,45.70; HRMS(ESI)calcd for C 23 H 15 F3N4O[M+H] + :421.1297.Found:421.1276.

[0196] Example 21: 4-(7-(Aminomethyl)-3-(4-fluorophenyl)quinoxalin-2-yl)benzonitrile (I-21)

[0197] Using 18e (100 mg, 0.22 mmol) as raw material, the preparation method is the same as I-1 to obtain yellow solid I-21 (35.21 mg) with a yield of 45.2% and a melting point of 77.2°C-78.9°C. 1 H NMR (500MHz, DMSO-d6): δ8.19–8.14(m,2H,ArH),7.95(dd,J=16.5,8.7Hz,3H,ArH),7.73(d,J=8 .0Hz,2H,ArH),7.58(dd,J=8.4,5.4Hz,2H,ArH),7.29(t,J=8.7Hz,2H,ArH),4.07(s,2H,-CH2-); 13 C NMR (126MHz, DMSO-d6): δ163.89,161.93,151.72,151.67,143.94,140.94,140.23,135.21,132.63 ,132.55,131.73,131.15,128.78,126.02,119.03,115.80,115.63,111.79,45.73; HRMS(ESI)calcd for C 22 H 15 FN4[M+H] + :355.1380.Found:355.1325.

[0198] Example 22: 4-(7-(Aminomethyl)-3-(4-pyridyl)quinoxalin-2-yl)benzonitrile (I-22)

[0199] Using 18f (100 mg, 0.23 mmol) as raw material, the preparation method is the same as I-1 to obtain yellow solid I-22 (31.77 mg) with a yield of 41.2% and a melting point of 170.9°C-172.4°C. 1 H NMR (500MHz, DMSO-d6): δ8.61(d,J=6.0Hz,2H,-CH=),8.22–8.12(m,2H,-CH=),7.95(dd,J=8.7,1.7Hz,1H,A rH),7.90(d,J=8.1Hz,2H,ArH),7.70(d,J=8.1Hz,2H,ArH),7.47(d,J=6.1Hz,2H,ArH),4.04(s,2H,-CH2-); 13C NMR (126MHz, DMSO-d6): δ151.70,150.46,150.08,146.28,143.30,141.28,140.18,132 .62,131.98,131.23,128.95,126.07,124.71,118.98,112.08,45.70; HRMS(ESI)calcd forC 21 H 15 N5[M+H] + :338.1406.Found:338.1439.

[0200] 1,2-Dimethoxy-4,5-dinitrobenzene (19a)

[0201] Add 10 ml of 65% concentrated nitric acid to a 50 ml single-necked bottle, place it in an ice bath, and slowly drop o-phenyl dimethyl ether (1 ml, 7.84 mmol) into it while stirring vigorously. 浓硝酸 :V 邻苯二甲醚 =10:1. After the addition, the temperature was raised to 80°C and heated in an oil bath for 1 h. The starting material disappeared after TLC detection (ethyl acetate: petroleum ether = 1:5). The reaction solution was cooled to room temperature and placed in an ice bath. A yellow precipitate was precipitated. The filter cake was washed with distilled water several times until it was neutral. The filter cake was purified by column chromatography using ethyl acetate: petroleum ether = 1:20 as the eluent to obtain a yellow solid 19a (1.60 g). The yield was 96.8%, MS [M+H] + :229.03.

[0202] 4,5-Dimethoxybenzene-1,2-diamine (20a)

[0203] 19a (1.5 g, 6.57 mmol) was added to a 150 ml single-necked bottle, dissolved in methanol (50 ml), and then 10% Pd / C (150 mg) was added. After the addition, H2 was introduced to exhaust the air three times. The reaction was carried out at room temperature for 1 hour. TLC detected the disappearance of the raw material (ethyl acetate: petroleum ether = 1:3). The reaction solution was filtered, and the filtrate was evaporated to remove the solvent under reduced pressure. The product did not need further purification and was directly used for the next step. MS [M+H] + :169.08.

[0204] 4-Ethylbenzonitrile (21a)

[0205] In a 120 ml pressure tube, 4-iodobenzonitrile (2.0 g, 8.73 mmol), tetrakis(triphenylphosphine)palladium (0.5 g, 0.43 mmol), and CuI (170 mg, 0.89 mmol) were added. The mixture was evacuated with N2 for 3 times. Et3N (18 ml) was added. The mixture was placed in an ice bath and trimethylethynylsilane (1.5 ml, 10.4 mmol) was slowly added dropwise. After the addition was completed, N2 was evacuated for 3 times. The mixture was heated to room temperature for 1 h. The raw material disappeared after TLC detection (ethyl acetate: petroleum ether = 1: 20), filtered, washed the filter cake with ether several times, the filtrate was dried, the crude product was redissolved with methanol (50 ml), K2CO3 (500 mg, 3.62 mmol) was added, reacted at room temperature for 1 h, TLC detection (ethyl acetate: petroleum ether = 1:12), water was added to the reaction solution until no precipitation occurred, filtered, the filter cake was removed, and the filter cake was purified by column chromatography, ethyl acetate: petroleum ether = 1:45 as eluent, to obtain a white solid 21a (601 mg), yield 54.2%, MS [M + H] + :128.03. 4-(p-Tolueneethynyl)benzonitrile (22a)

[0206] 21a (600 mg, 4.72 mmol), p-iodotoluene (857 mg, 3.93 mmol), palladium acetate (9 mg, 0.0401 mmol), CuI (7 mg, 0.0367 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (23 mg, 0.0397 mmol) and Cs2CO3 (2.60 g, 7.98 mmol) were added to a 150 ml single-necked bottle, followed by DMF (50 ml), and the reaction was heated in an oil bath at 60°C for 0.5 h. The raw material disappeared after TLC detection (ethyl acetate: petroleum ether = 1:20). Excess water was added to the reaction solution until no more precipitation occurred, and the filter cake was purified by column chromatography with ethyl acetate: petroleum ether = 1:180 as the eluent to obtain 22a (935 mg) as a white solid with a yield of 91.2%, MS [M+H] + :218.08.

[0207] 4-(2-Oxo-2-(p-methylphenyl)acetyl)benzonitrile (23a)

[0208] 22a (900 mg, 4.15 mmol) was added to a 50 ml single-necked bottle, dissolved with DMSO (20 ml), and then I2 (210 mg, 0.829 mmol) was added. The reaction was heated in an oil bath at 130°C for 3 days. The starting material disappeared after TLC detection (ethyl acetate: petroleum ether = 1:15). Excess water was added to the reaction solution until no more precipitation occurred. The filter cake was purified by column chromatography using ethyl acetate: petroleum ether = 1:60 as the eluent to obtain a white solid 23a (776 mg). The yield was 75.2%, MS [M+H] +:250.07.

[0209] 4-(6,7-Dimethoxy-3-(p-tolyl)quinoxalin-2-yl)benzonitrile (24a)

[0210] 23a (700 mg, 2.81 mmol) and crude product 20a were added to a 150 ml single-necked bottle, dissolved with methanol (50 ml), evacuated with N2 for 3 times, reacted at room temperature for 1 hour, and the raw material disappeared after TLC detection (ethyl acetate: petroleum ether = 1:3). Excess water was added to the reaction solution until no more precipitation occurred, filtered, and the filter cake was purified by column chromatography, using ethyl acetate: petroleum ether = 1:9 as the eluent, to obtain a light yellow solid 24a (752 mg), with a yield of 70.2%, MS [M+H] + :382.14.

[0211] Example 23: 4-(6,7-dihydroxy-3-(p-tolyl)quinoxalin-2-yl)benzonitrile (I-23)

[0212] 24a (700 mg, 1.84 mmol) was added to a 50 ml single-necked bottle, dissolved in toluene (20 ml), and then AlCl3 (979 mg, 7.34 mmol) was added. The reaction was heated in an oil bath at 110°C for 6 h. The raw material disappeared after TLC detection (ethyl acetate: petroleum ether = 1:2). The reaction solution was poured into ice water, the pH was adjusted to 1 with 2N HCl, extracted with EA, the organic layer was collected, dried over anhydrous Na2SO4, filtered, the filtrate was evaporated under reduced pressure to remove the solvent, and purified by column chromatography to obtain a brown solid (227 mg), with a yield of 35.0%, and a melting point of 155.8°C-157.2°C. 1 HNMR (500MHz, DMSO-d6): δ10.61(s,2H,-OH),7.81(d,J=7.9Hz,2H,ArH),7.59(d,J=7.9 Hz,2H,ArH),7.29(d,J=8.7Hz,4H,ArH),7.15(d,J=7.8Hz,2H,ArH),2.31(s,3H,-CH3); 13 CNMR (126MHz, DMSO-d6): δ152.21,151.78,149.58,147.90,144.86,138.38,138.28,137.80,13 6.50,132.39,131.00,130.13,129.21,119.18,111.01,109.61,109.51,21.29; HRMS(ESI)calcd for C 22 H 15 N3O2[M+H] + :354.1243.Found:354.1270.

[0213] (R)-tert-Butyl (1-(4-(2-(4-cyanophenyl)-2-oxoacetamido)-3-nitrophenyl)pyrrolidin-3-yl)carbamate (25a)

[0214] Using 6a (2 g, 11.42 mmol) and 3d (2.9 g, 9.14 mmol) as raw materials, the preparation method was the same as 7a to obtain yellow solid 25a (1.02 g) with a yield of 35.1%, MS [M+H] + :480.17.

[0215] (R)-tert-Butyl (1-(3-(4-cyanophenyl)-2-oxo-1,2-dihydroquinoxalin-6-yl)pyrrolidin-3-yl)carbamate (26a)

[0216] Using 25a (1.50 g, 3.13 mmol) as starting material, the preparation method was the same as 8a to obtain yellow solid 26a (459 mg) with a yield of 34.2%, MS [M+H] + :432.19.

[0217] (R)-tert-Butyl (1-(3-(4-cyanophenyl)-2-(p-tolyl)quinoxalin-6-yl)pyrrolidin-3-yl)carbamate (27a)

[0218] Using 26a (100 mg, 0.22 mmol) and 4-methylphenylboronic acid (68 mg, 0.45 mmol) as raw materials, the preparation method was the same as 9a to obtain yellow solid 27a (109 mg) with a yield of 90.0%, MS [M+H] + :522.23.

[0219] Example 24: (R)-4-(7-(3-aminopyrrolidin-1-yl)-3-(p-tolyl)quinoxalin-2-yl)benzonitrile (I-24)

[0220] Using 27a (100 mg, 0.19 mmol) as raw material, the preparation method is the same as I-1 to obtain yellow solid I-24 (42 mg) with a yield of 52% and a melting point of 191.8°C-194.2°C. 1H NMR (500MHz, DMSO-d6): δ7.92 (d, J=9.3Hz, 1H, ArH), 7.83 (d, J=8.4Hz, 2H, ArH), 7.62 (d,J=8.3Hz,2H,ArH),7.38(dd,J=9.3,2.7Hz,1H,ArH),7.29(d,J=8.1Hz,2H,ArH),7. 15(d,J=7.9Hz,2H,ArH),6.83(d,J=2.6Hz,1H,ArH),3.68–3.44(m,4H,-CH2-×2),3.1 3–3.10(m,1H,-CH-),2.31(s,3H,-CH3),2.15–1.80(m,2H,-CH2-),1.23(s,2H,-NH2); 13 C NMR (126MHz, DMSO-d6): δ151.39,149.05,147.32,144.84,143.03,138.07,136.39,135.05,132.41,131.00,1 30.04,129.85,129.18,121.25,119.12,111.34,103.35,56.49,51.36,46.71,34.52,21.27; HRMS(ESI)calcd for C 21 H 15 N5[M+H] + :406.2032.Found:406.2047.

[0221] For the structure determination of the above examples, the melting point was determined using a b-type melting point tube, the medium was methyl silicone oil, and the thermometer was not calibrated; 1 HNMR was measured using a JEOL FX90Q Fourier transform nuclear magnetic resonance instrument and a BRUKER nuclear magnetic resonance instrument (TMS internal standard); MS was measured using a Nicolet 2000 Fourier transform mass spectrometer and a MAT-212 mass spectrometer.

[0222] Pharmacological activity test

[0223] Activity test

[0224] 1. Determination of the inhibitory activity of LSD1

[0225] The compound was prepared into a DMSO solution with a stock concentration of 20 mmol / L and stored in the dark for future use.

[0226] (1) First prepare 1× buffer;

[0227] (2) Compound concentration preparation: Compound test IC 50The final concentration was 10 μmol / L, 3-fold dilution, 8 concentrations, and each concentration was tested in a single well. The inhibition rate test concentration was 1 μmol / L, and the test was repeated. The positive control compound ORY-1001 test final concentration was 100 nmol / L, 3-fold dilution, 10 concentrations, and each concentration was tested in repeated wells. Diluted to the corresponding 1000-fold final concentration solution in the 384-well Source plate, and then transferred 10nl to the 384-well reaction plate for testing using Echo550. 10 nmol / L of 100% DMSO was transferred to the Min and Max wells.

[0228] (3) Prepare 2× enzyme solution using 1× reaction solution.

[0229] (4) Use 1× reaction solution to prepare 2× substrate mixed solution.

[0230] (5) Add 5 μl of 2× enzyme solution to each well; add 5 μl of 1× reaction solution to the Min well, centrifuge at 1000 rpm for 1 min, and incubate at room temperature for 15 min.

[0231] (6) Add 5 μl of 2× substrate mixture solution to each well of the reaction plate to start the reaction, centrifuge at 1000 rpm for 1 min, and incubate at room temperature for 60 min.

[0232] (7) Add 10 μl of detection solution to each well, centrifuge at 1000 rpm for 1 min, and incubate at room temperature for 60 min.

[0233] (8) Use EnVision to read the signal Intensity (665nm) / Intensity (615nm).

[0234] 2. Data Analysis

[0235] Calculation formula: Inhibition percentage (%) = (maximum value - signal value) / (maximum value - minimum value) × 100

[0236] Fitting the dose-effect curve: With the logarithm of the concentration as the X-axis and the percentage inhibition rate as the Y-axis, the log(inhibitor) vs.response-Variable slope formula of the analysis software GraphPad Prism 5 was used to fit the dose-effect curve, thereby obtaining the IC value of each compound on the enzyme activity. 50 value.

[0237] 3. MTS assay to detect cell activity

[0238] TS is a new MTT analogue, the full name is 3-(4,5-dimethylthiazol-2-yl)-5(3-carbocymethoxyphenyl)-2-(4-sulfopheny)-2H-tetrazolium, a yellow dye. Succinate dehydrogenase in the mitochondria of living cells can metabolize and reduce MTS to generate soluble formazan compounds, and the content of formazan can be measured at 490nm using an enzyme marker. Under normal circumstances, the amount of formazan generated is proportional to the number of living cells, so the number of living cells can be inferred based on the optical density OD value.

[0239] 4. Experimental Methods

[0240] (1) Inoculation of cells: Prepare a single cell suspension in a culture medium (DMEM or RMPI1640) containing 10% fetal bovine serum, and inoculate 3,000 to 15,000 cells per well into a 96-well plate. The volume of each well is 100 μl. The cells are inoculated and cultured 12 to 24 hours in advance. (2) Add the solution of the compound to be tested: Dissolve the compound in DMSO, and screen the compound at a concentration of 40 μmol / L. The final volume of each well is 200 μl. Three replicate wells are set for each treatment. The compound is screened at a concentration of 40, 8, 1.6, 0.32, 0.064, 0.0128, 0.00256, and 0.000512 μmol / L. The specific screening concentrations of each compound are detailed in the rescreening data. The final volume of each well is 200 μl. Three replicate wells are set for each treatment.

[0241] (3) Color development: After culturing at 37°C for 48 hours, discard the culture medium in the wells for adherent cells and add 20 μl of MTS solution and 100 μl of culture medium to each well; discard 100 μl of culture supernatant for suspended cells and add 20 μl of MTS solution to each well; set up 3 blank replicate wells (a mixture of 20 μl MTS solution and 100 μl culture medium) and continue incubating for 2 to 4 hours to allow the reaction to proceed fully before measuring the absorbance value.

[0242] (4) Colorimetry: Select 492 nm wavelength and use a multifunctional microplate reader (MULTISKAN FC) to read the light absorption value of each well and record the results. After data processing, draw a cell growth curve with concentration as the horizontal axis and cell survival rate as the vertical axis. Calculate the IC of the compound using the two-point method (Reed and Muench method) 50 value.

[0243] (5) Positive control compounds: Two positive control compounds, cisplatin (DDP) and paclitaxel (Taxol), were set in each experiment. The cell growth curve was drawn with the concentration as the horizontal axis and the cell survival rate as the vertical axis. The IC of the compound was calculated using the two-point method (Reed and Muench method). 50 value.

[0244] 5. Target compound activity test results

[0245] Based on the designed synthetic route, we synthesized target compounds Ⅰ-1 to Ⅰ-24, and used ORY-1001() as a positive control compound to test the inhibition rate of target compounds on LSD1. For compounds with better activity, their IC values ​​on LSD1 were measured. 50 In addition, the anti-proliferative activity of the target compound on tumor cells was also tested, and the test results are as follows:

[0246] Table 1 Activity test results of series I target compounds against LSD1

[0247]

[0248]

[0249] a Inhibition rate at a compound concentration of 1 μmol / L; b A: inhibition rate>80%, B: 79%>inhibition rate>50%, C: 49%>inhibition rate; c The highest concentration of the compound was 10 μmol / L; d D:IC 50 <1μmol / L, E: 5μmol / L>IC 50 >1μmol / L, F:IC 50 >5μmol / L; ND not tested

[0250] Table 2 Activity test results of target compounds I-8 and I-18 on tumor cells

[0251]

[0252] d D:IC 50 <1μmol / L, E: 5μmol / L>IC 50 >1μmol / L, F:IC 50 >5 μmol / L;

[0253] Studies have shown that this class of compounds has significant inhibitory activity against LSD1. The inhibitory activity of the optimal compound Ⅰ-8 against LSD1 is 0.204μmol / L, and it also exhibits significant anti-proliferative activity against liver cancer SMMC-7721. This class of compounds has a novel structure and strong inhibitory activity against LSD1. It is a new and potent LSD1 inhibitor. The discovery of this class of compounds has laid the foundation for the development of new anti-tumor drugs targeting LSD1.

Claims

1. A compound having the following structural formula (I) or a pharmaceutically acceptable salt thereof: in: R 1 and R 2 Independently selected from hydrogen, -OH, substituted or unsubstituted 5-10 membered heterocyclic group or -N(R), the heterocyclic group contains 1-3 heteroatoms of N, O or S, substituted or unsubstituted means that the corresponding functional group has or does not have a substituent, the substituent is C1-C6 alkyl, C1-C6 alkoxy, amino, C1-C6 alkylamino; in -N(R), each R is independently selected from hydrogen, C1-C7 fatty primary amine, C1-C7 fatty secondary amine, C1-C7 fatty tertiary amine; W 1 and W 2 Independently selected from N, CH, CF, C-NH2, C-CH3; R 3 Selected from substituted or unsubstituted 5-10 membered aryl, 5-10 membered heteroaryl, C3-C6 cycloalkyl, C1-C6 alkyl, 5-10 membered heterocyclic group, substituted or unsubstituted means that the corresponding functional group has or does not have a substituent, and the substituent is C1-C6 alkyl, C1-C6 alkoxy, halogen, halogenated C1-C6 alkyl, nitro, amino, 5-10 membered aryl.

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: R 1 and R 2 independently selected from substituted or unsubstituted 5-6 membered heterocyclic group or -N(R 4 )2, the heterocyclic group contains 1-2 N heteroatoms, and the substituent is C1-C6 alkyl, C1-C6 alkoxy, amino, C1-C6 alkylamino; -N(R 4 )2, each R 4 are independently selected from propylamine, N-methylpropylamine, N,N-dimethylpropylamine; R 3 Selected from substituted or unsubstituted 5-10 membered aryl, 5-10 membered heteroaryl, C3-C6 cycloalkyl, C1-C6 alkyl, 5-10 membered heterocyclic group, the substituent is C1-C3 alkyl, C1-C3 alkoxy, halogen, halogenated C1-C3 alkyl, amino.

3. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The R 1 and R 2 Independently selected from hydrogen, methyl, methoxy, hydroxyl, substituted or unsubstituted groups: W 1 and W 2 Independently selected from N, CH, CF, C-CH3; R 3 is optionally substituted or unsubstituted 5-10 membered aryl or 5-10 membered heteroaryl, and the substituent is C1-C3 alkyl, C1-C3 alkoxy, halogen, halogenated C1-C3 alkyl, or amino.

4. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The R 1 and R 2 Independently selected from hydrogen, methyl, methoxy, hydroxyl, substituted or unsubstituted groups: W 1 and W 2 Independently selected from N, CH, CF, C-CH3; R 3 is optionally substituted or unsubstituted as follows: The substituents are C1-C3 alkyl, C1-C3 alkoxy, halogen, halogenated C1-C3 alkyl, and amino.

5. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The compound is selected from: (S)-4-(7-(3-aminopyrrolidin-1-yl)-3-(4-methoxyphenyl)quinoxalin-2-yl)benzonitrile (I-1) (S)-4-(7-(3-aminopyrrolidin-1-yl)-3-(4-methylphenyl)quinoxalin-2-yl)benzonitrile (I-2) (S)-4-(7-(3-aminopyrrolidin-1-yl)-3-(4-(trifluoromethyl)phenyl)quinoxalin-2-yl)benzonitrile (I-3) (S)-4-(7-(3-aminopyrrolidin-1-yl)-3-(4-(trifluoromethoxy)phenyl)quinoxalin-2-yl)benzonitrile (I-4) (S)-4-(7-(3-aminopyrrolidin-1-yl)-3-(4-fluorophenyl)quinoxalin-2-yl)benzonitrile (I-5) (S)-4-(7-(3-aminopyrrolidin-1-yl)-3-(4-pyridyl)quinoxalin-2-yl)benzonitrile (I-6) (S)-4-(7-(3-aminopiperidin-1-yl)-3-(4-methoxyphenyl)quinoxalin-2-yl)benzonitrile (I-7) (S)-4-(7-(3-aminopiperidin-1-yl)-3-(4-methylphenyl)quinoxalin-2-yl)benzonitrile (I-8) (S)-4-(7-(3-aminopiperidin-1-yl)-3-(4-trifluoromethylphenyl)quinoxalin-2-yl)benzonitrile (I-9) (S)-4-(7-(3-aminopiperidin-1-yl)-3-(4-trifluoromethoxyphenyl)quinoxalin-2-yl)benzonitrile (I-10) (S)-4-(7-(3-aminopiperidin-1-yl)-3-(4-fluorophenyl)quinoxalin-2-yl)benzonitrile (I-11) (S)-4-(7-(3-aminopiperidin-1-yl)-3-(4-pyridine)quinoxalin-2-yl)benzonitrile (I-12) 4-(7-(3-aminopiperazin-1-yl)-3-(4-methoxyphenyl)quinoxalin-2-yl)benzonitrile (I-13) 4-(7-(3-aminopiperazin-1-yl)-3-(4-methylphenyl)quinoxalin-2-yl)benzonitrile (I-14) 4-(7-(3-aminopiperazin-1-yl)-3-(4-fluorophenyl)quinoxalin-2-yl)benzonitrile (I-15) 4-(7-(3-aminopiperazin-1-yl)-3-(4-pyridyl)quinoxalin-2-yl)benzonitrile (I-16) 4-(7-(Aminomethyl)-3-(4-methoxyphenyl)quinoxalin-2-yl)benzonitrile (I-17) 4-(7-(Aminomethyl)-3-(4-methylphenyl)quinoxalin-2-yl)benzonitrile (I-18) 4-(7-(Aminomethyl)-3-(4-(trifluoromethyl)phenyl)quinoxalin-2-yl)benzonitrile (I-19) 4-(7-(Aminomethyl)-3-(4-(trifluoromethoxy)phenyl)quinoxalin-2-yl)benzonitrile (I-20) 4-(7-(Aminomethyl)-3-(4-fluorophenyl)quinoxalin-2-yl)benzonitrile (I-21) 4-(7-(Aminomethyl)-3-(4-pyridyl)quinoxalin-2-yl)benzonitrile (I-22) 4-(6,7-Dihydroxy-3-(p-tolyl)quinoxalin-2-yl)benzonitrile (I-23) (R)-4-(7-(3-aminopyrrolidin-1-yl)-3-(p-tolyl)quinoxalin-2-yl)benzonitrile (I-24) 6. The compound according to any one of claims 1 to 5 or a pharmaceutically acceptable salt thereof, wherein the pharmaceutically acceptable salt is an acid addition salt formed by the compound of formula (I) and the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid or succinic acid, fumaric acid, salicylic acid, phenylacetic acid or mandelic acid.

7. A method for preparing the compound according to any one of claims 1 to 6, characterized in that: R 2 is hydrogen, R 1 For other functional groups except -OH, comprising the following steps: (1) Under the action of SeO2, the reaction generates (2) Amine intermediates Condensation (3) The nitro group is reduced and the cyclization is carried out simultaneously to obtain (4) and Response (5) Hydrolysis under acidic conditions yields Among them, R 5 Contains Boc, R 1 To R 5 The Boc in is replaced by H.

8. The method according to claim 7, characterized in that In step (2), the nitro group is reduced and cyclization occurs simultaneously under the action of hydrogen / palladium carbon or iron powder / HCl or hydrazine hydrate.

9. A pharmaceutical composition comprising the compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof.

10. Use of the compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 9 in the preparation of a medicament for preventing or treating a clinical disorder associated with LSD1.

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