4-Amino-7-azaindole compounds, preparation method and application thereof

Abstract

The present invention provides a 4-amino-7-azaindole compound, a preparation method and application thereof, and belongs to the technical field of compound synthesis. The 4-amino-7-azaindole compound or its stereoisomers and pharmaceutically acceptable salts have the structural formula: The compound can effectively inhibit the activity of ULKs kinase, especially the activity of ULK1 kinase and ULK2 kinase, and is expected to be a new inhibitor of ULK1 kinase and ULK2 kinase.

Description

Technical Field

[0001] The invention belongs to the technical field of compound synthesis, and specifically relates to 4-amino-7-azaindole compounds, preparation methods and applications thereof. Background Art

[0002] Autophagy is a cellular catabolic process that plays an important role in cell growth, differentiation, homeostasis and cell survival under conditions of nutrient deficiency. It is an important self-protection and defense mechanism of the body. Autophagy specifically refers to the double-layer membrane structure in the cell that encapsulates part of the cytoplasm, damaged proteins, and aging or damaged organelles such as mitochondria, Golgi bodies, and endoplasmic reticulum to form autophagosomes, which are delivered to lysosomes and fuse with them to form autophagolysosomes. The contents are hydrolyzed by proteolytic enzymes for digestion and degradation to meet the cell's own metabolism and renewal.

[0003] The process of cellular autophagy usually originates from various stress conditions, such as starvation, hypoxia, oxidative stress, protein aggregation, endoplasmic reticulum stress, etc. At present, the source of the membrane involved in the formation of phagosomes is still unclear, and it may be the endoplasmic reticulum, mitochondria, Golgi apparatus, and plasma membrane. After the phagosome is formed, it expands and separates damaged organelles (including the endoplasmic reticulum, ribosomes, and mitochondria, etc.) and misfolded proteins. As the phagosome expands, the cell membrane bends and eventually forms a double-layer membrane structure autophagosome. Finally, the autophagosome fuses with the lysosome to form an autophagosome, and the substances in the autophagosome are degraded by the acid hydrolases in the lysosome. In addition, the nutrients produced during the degradation process are released back to the cytoplasm and reused by the cell. Although autophagy enables cells to survive the stress conditions faced by different stages of cancer, excessive autophagy and autophagic cell death may kill cancer cells and limit their progression and metastasis.

[0004] ULK is a serine / threonine protein kinase, including 5 isoforms, namely ULK1-4 and STK36. ULK1 forms a protein complex with the autophagy proteins mATG13, FIP200 and ATG101 to regulate the initiation of autophagy. It has been shown that inactive mutants of ULK kinase can block the typical initiation of autophagy, indicating that small molecule inhibitors of ULK can block autophagy. In addition, when ULK1 is inhibited, ULK2 can compensate for the function of ULK1 in autophagy regulation, so when designing small molecule inhibitors, ULK1 and ULK2 should be targeted at the same time.

[0005] At present, inhibitors targeting ULK1 / 2 include SBI-0206965, ULK100, ULK101, MRT-67307, MRT-68921, ERAS-5, ENV-201 and DCC-3116, most of which are in the preclinical research stage, and only DCC-3116 has entered early clinical research. In summary, there is still a huge market demand for further development of new ULK1 / 2 inhibitors. Summary of the invention

[0006] Based on the deficiencies in the prior art, the present invention aims to provide a 4-amino-7-azaindole compound and a preparation method thereof, which can effectively inhibit the activity of ULKs kinases, especially the activity of ULK1 kinase and ULK2 kinase, and is expected to be a new inhibitor of ULK1 kinase and ULK2 kinase.

[0007] In order to achieve the above object, the present invention adopts the following technical solutions:

[0008] In one aspect, the present invention provides a 4-amino-7-azaindole compound or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, the structural formula of which is shown in the following formula (1):

[0009] (1)

[0011] in,

[0012] X is selected from -CH2- or a bond,

[0013] R 1 Selected from , , or ;

[0014] R 2 Selected from hydrogen;

[0015] or R 1 , R 2 The carbonyl group and nitrogen atom connected to it form or , The carbon of can be replaced by an -O- or -NH-;

[0016] R 3 , R 4 independently selected from hydrogen, fluorine or C 1-3 alkyl;

[0017] R 5 Selected from hydrogen;

[0018] or R 4, R 5 The connected methylene and nitrogen atoms form , or ;

[0019] R 6 is selected from halogen, trifluoromethyl or ethynyl;

[0020] R 7 and R 8 At least one is hydrogen and the other is selected from C 4-6 Heterocyclic group, C 5-7 Bridged heterocyclic group, C 6-8 fused heterocyclic group or cyano group, said C 4-6 Heterocyclic group, C 5-7 Bridged heterocyclic group, C 6-8 The fused heterocyclic group may be independently selected from C 1-3 Alkyl, cyclopropyl, fluorine, one or two or three substituents, containing up to three identical or different heteroatoms selected from N, O, S;

[0021] n is selected from 1, 2, and 3.

[0022] Preferably, the 4-amino-7-azaindole compound or its stereoisomers or pharmaceutically acceptable salts has a structural formula as shown in the following formula (2):

[0023] (2)

[0025] in,

[0026] R 1 Selected from , , or ;

[0027] R 2 Selected from hydrogen;

[0028] or R 1 , R 2 The carbonyl group and nitrogen atom connected to it , , , , , or ;

[0029] R 3 , R 4 independently selected from hydrogen, fluorine or C 1-3 alkyl;

[0030] R6 is selected from halogen, trifluoromethyl or ethynyl;

[0031] R 7 and R 8 At least one of them is hydrogen and the other one is selected from , , , , , , or cyano.

[0032] More preferably, the 4-amino-7-azaindole compound or its stereoisomers or pharmaceutically acceptable salts has a structural formula as shown in the following formula (3):

[0033] (3)

[0035] in,

[0036] R 4 Selected from hydrogen, fluorine or C 1-3 The alkyl group;

[0037] R 6 is selected from trifluoromethyl or ethynyl;

[0038] R 7 and R 8 At least one of them is hydrogen and the other one is selected from , , , , , or cyano.

[0039] As some preferred embodiments, the 4-amino-7-azaindole compound or its stereoisomers or pharmaceutically acceptable salts is one of the following compounds:

[0040]

[0041]

[0042]

[0043]

[0044] On the other hand, the present invention also provides a pharmaceutical composition, which comprises the above-mentioned 4-amino-7-azaindole compound or its stereoisomer, a pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier. The pharmaceutical carrier mentioned herein refers to a pharmaceutical carrier commonly used in the pharmaceutical field; the pharmaceutical composition can be prepared according to methods well known in the art. The 4-amino-7-azaindole compound or its stereoisomer, a pharmaceutically acceptable salt, and one or more pharmaceutically acceptable solid or liquid excipients and / or adjuvants described in the present invention can be combined to prepare any dosage form suitable for human or animal use.

[0045] The 4-amino-7-azaindole compounds or their stereoisomers, pharmaceutically acceptable salts or pharmaceutical compositions containing them described in the present invention can be administered in unit dosage forms, and the administration route can be enteral or parenteral, such as oral, intravenous, intramuscular, subcutaneous, nasal, oral mucosa, eyes, lungs and respiratory tract, skin, vagina, rectum, etc.

[0046] The dosage form can be a liquid dosage form, a solid dosage form or a semisolid dosage form. Liquid dosage forms can be solutions (including true solutions and colloidal solutions), emulsions (including o / w types, w / o types and multiple emulsions), suspensions, injections (including water injections, powder injections and infusions), eye drops, nasal drops, lotions and liniments, etc.; solid dosage forms can be tablets (including ordinary tablets, enteric-coated tablets, lozenges, dispersible tablets, chewable tablets, effervescent tablets, orally disintegrating tablets), capsules (including hard capsules, soft capsules, enteric-coated capsules), granules, powders, micropills, dropping pills, suppositories, films, patches, aerosols (powders), sprays, etc.; semisolid dosage forms can be ointments, gels, pastes, etc.

[0047] The 4-amino-7-azaindole compounds or their stereoisomers and pharmaceutically acceptable salts described in the present invention can be made into common preparations, sustained-release preparations, controlled-release preparations, targeted preparations and various microparticle drug delivery systems.

[0048] The 4-amino-7-azaindole compound or its stereoisomers and pharmaceutically acceptable salts in the pharmaceutical composition can be used alone or mixed with other drugs; the other drugs include chemotherapeutic drugs;

[0049] The chemotherapy drug is selected from: one or more of trametinib, selumetinib, cobimetinib, bimetinib, uritinib, SCH772984, LY3214996, lavotinib, VX-11e, LY3009120, nabolafenib, RAF709, dabrafenib, vemurafenib, sotolacib and adagracib;

[0050] In one aspect, the present invention also provides the use of the above-mentioned 4-amino-7-azaindole compound or its stereoisomer, pharmaceutically acceptable salt or the pharmaceutical composition in the preparation of a drug for treating diseases caused by abnormal activation of ULK kinase;

[0051] The disease is an autoimmune disease, cardiovascular disease, metabolic disease, liver disease, neurodegenerative disease, infectious disease or cancer; the cancer is metastatic cancer; the metastatic cancer is gastrointestinal stromal tumor, esophageal cancer, gastric cancer, melanoma, glioma, glioblastoma, ovarian cancer, bladder cancer, pancreatic cancer, prostate cancer, lung cancer, breast cancer, kidney cancer, liver cancer, osteosarcoma, multiple myeloma, cervical cancer, bone metastasis, papillary thyroid cancer, non-small cell lung cancer or colorectal cancer. Detailed description of the invention:

[0053] Various aspects and features of the present invention are further described below.

[0054] All documents cited in the present invention are incorporated herein by reference in their entirety, and if the meanings expressed in these documents are inconsistent with the present invention, the description of the present invention shall prevail. In addition, the various terms and phrases used in the present invention have the general meanings known to those skilled in the art. Even so, the present invention still hopes to provide a more detailed description and explanation of these terms and phrases herein. If the terms and phrases mentioned are inconsistent with the known meanings, the meanings described in the present invention shall prevail. Below are the definitions of various terms used in the present invention, which apply to the terms used in the entire specification of this application, unless otherwise specified in specific circumstances.

[0055] The following provides definitions of various groups in the compounds of the present invention, which are used uniformly in the specification and claims unless otherwise defined.

[0056] As mentioned in the present invention, the term "alkyl" refers to an alkyl group having a specified number of carbon atoms, which can be a straight chain or branched chain alkyl group, for example, the "C 1-3 "alkyl" includes methyl, ethyl, propyl and isopropyl; for example, when referring to "C 4-6 When the term "heterocyclic group" is used, it means a non-aromatic heterocyclic ring having 4, 5 or 6 carbon atoms, including a saturated heterocyclic ring and a non-aromatic unsaturated heterocyclic ring with a double bond. For example, when a "bridged (fused) heterocyclic group" is mentioned, it also includes a saturated bridged (fused) heterocyclic ring and a non-aromatic unsaturated bridged (fused) heterocyclic ring with a double bond.

[0057] As mentioned in the present invention, the term "isomer" means that the compounds of the present invention may exist in specific geometric or stereoisomer forms. In the present invention, isomers mainly refer to R 3 , R 4 For different substituents, there will be (R )-and( S )-enantiomer, racemate, R 4 , R 5 When it forms a five-membered ring or a six-membered ring with the connected methylene and nitrogen atom, ( R )-and( S )-enantiomers, racemates, all of these isomers and mixtures thereof are included within the scope of the present invention.

[0058] Compared with the prior art, the present invention has the following beneficial effects:

[0059] The present invention provides a 4-amino-7-azaindole compound and a preparation method thereof. The compound can effectively inhibit the activity of ULKs kinases, especially has good inhibition on the activities of ULK1 kinase and ULK2 kinase. DETAILED DESCRIPTION

[0060] The following examples further illustrate the present invention but are not intended to limit the present invention in any way.

[0061] Embodiment 1: N -((1-(5-bromo-2-(4-cyanophenyl)-7-azaindol-4-yl)piperidin-3-yl)methyl)cyclobutylcarboxamide (B001)

[0062]

[0063] Step 1: Synthesis of intermediate 1-2

[0064] Dissolve cyclobutylcarboxylic acid (300.3 mg, 3.0 mmol) in 15 mL acetonitrile, add EDCI (345.0 mg, 1.8 mmol), HOBT (285.0 mg, 2.1 mmol) and DIPEA (582.0 mg, 4.5 mmol) in turn, stir at room temperature for 30 minutes, add 1-1 (321.5 mg, 1.5 mmol), and stir at room temperature overnight. When the reaction is complete, add 20 mL of water to quench the reaction, extract with dichloromethane (30 mL × 3), combine the organic phases, dry over anhydrous sodium sulfate, filter, and spin dry. The crude product was purified by column chromatography (PE:EA = 20:1~10:1) to obtain 355.6 mg (1-2) of solid. ESI-MS: m / z = 297 [M+H] + .

[0065] Step 2: Synthesis of intermediate 1-3

[0066] Add 5 mL of hydrochloric acid-ethyl acetate solution (2M) to a reaction bottle containing 1-2 (355.6 mg, 1.2 mmol) and react at room temperature for 3 hours. When the reaction is complete, spin dry the solvent to obtain 409.1 mg of a white solid (1-3 hydrochloride), which is used directly in the next step. ESI-MS: m / z = 197 [M+H] + .

[0067] Step 3: Synthesis of intermediate 1-5

[0068] 1-4 (1.07 g, 7.0 mmol) was added to 20 mL of anhydrous THF. o C. NaH (60%) was slowly added w ,336.0 mg, 8.4 mmol), stirred for 20 minutes, SEM-Cl (1.40 g, 8.4 mmol) was added dropwise, and the reaction was allowed to react at room temperature for 3 hours. After the reaction was complete, 40 ml of water was added to quench the reaction, extracted with ethyl acetate (30 mL × 5), washed with saturated brine (50 ml × 2), dried over anhydrous sodium sulfate, filtered, and dried. The crude product was purified by column chromatography (PE:EA = 100:0~10:1) to obtain 1.43 g of solid (1-5). ESI-MS: m / z = 283 [M+H] + .

[0069] Step 4: Synthesis of Intermediate 1-6

[0070] 1-5 (1.43 g, 5.0 mmol) was dissolved in 25 mL of anhydrous tetrahydrofuran. o 2.5 mL of n-butyl lithium in n-hexane solution (2.5 M) was added dropwise at 4 °C, and triisopropyl borate (1.10 g, 6.0 mmol) was added after stirring for 30 minutes. The mixture was reacted at room temperature for 3 hours. o 10 mL of saturated ammonium chloride solution was added at 40 °C to quench the reaction. After returning to room temperature, the mixture was extracted with dichloromethane (50 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and dried by spin drying. The crude product was purified by column chromatography (PE:EA = 10:1~4:1) to obtain 621.9 mg of solid (1-6). ESI-MS: m / z = 327 [M+H] + .

[0071] Step 5: Synthesis of Intermediate 1-7

[0072] 1-6 (440 mg, 1.35 mmol), 4-iodobenzonitrile (370 mg, 1.61 mmol) and potassium carbonate (372 mg, 2.70 mmol) were dissolved in 35 mL of a mixed solvent of dioxane / water (v / v = 4 / 1), Pd(PPh3)4 (156 mg, 0.135 mmol) was added, nitrogen was replaced, and the mixture was heated at 70 °C. o C for 2 hours. After the reaction is complete, add 30 mL of water to dilute the reaction, extract with dichloromethane (50 mL × 3), combine the organic phases, dry over anhydrous sodium sulfate, filter, and spin dry. The crude product is purified by column chromatography (PE:EA = 20:1~10:1) to obtain 419.5 mg of solid (1-7). ESI-MS: m / z = 384 [M+H] + .

[0073] Step 6: Synthesis of Intermediate 1-8

[0074] Dissolve 1-7 (400 mg, 1.04 mmol) in 5 mL of dioxane, add 1-3 (305.8 mg, 1.56 mmol), Pd2(dba)3 (95.2 mg, 0.104 mmol), BINAP (129.6 mg, 0.208 mmol) and sodium tert-butoxide (249.6 mg, 2.6 mmol), replace nitrogen, and heat at 120 °C. o C for 17 hours. When the reaction is complete, add 10 mL of water to dilute the reaction, extract with dichloromethane (20 mL × 3), combine the organic phases, dry over anhydrous sodium sulfate, filter, and spin dry. The crude product is purified by column chromatography (PE:EA = 10:1~2:1~0:1) to obtain 133.3 mg of solid (1-8). ESI-MS: m / z = 544 [M+H] + .

[0075] Step 7: Synthesis of Intermediate 1-9

[0076] Dissolve 1-8 (133.3 mg, 0.25 mmol) in 1 mL of dichloromethane, add NBS (51.7 mg, 0.28 mmol), and react at room temperature for 2 hours. When the reaction is complete, add 5 mL of water to dilute the reaction, extract with dichloromethane (10 mL × 3), combine the organic phases, dry over anhydrous sodium sulfate, filter, and spin dry. The crude product was purified by preparative thin layer chromatography (PE:EA = 2:1) to obtain 40.0 mg of solid (1-9). ESI-MS: m / z = 622 [M+H] + .

[0077] Step 8: Synthesis of B001

[0078] Dissolve 1-9 (40.0 mg, 0.064 mmol) in 0.5 mL of dichloromethane, add 2.0 mL of trifluoroacetic acid, and react at room temperature for 8 hours. After the reaction is complete, add saturated sodium bicarbonate aqueous solution to adjust the reaction solution to weak alkalinity, extract with ethyl acetate (10 ml × 6), combine the organic phases, wash with saturated brine (20 ml × 2), dry with anhydrous sodium sulfate, filter, and spin dry. The crude product is purified by preparative thin layer chromatography (PE:EA = 1:1) to obtain 11.3 mg of solid (B001).

[0079] 1 H NMR (400 MHz, DMSO- d 6) δ 12.35 (s, 1H), 8.15 (s, 1H), 8.12 (d, J =8.4 Hz, 2H), 7.88 (d, J = 8.4 Hz, 2H), 7.73 (t, J = 6.0 Hz, 1H), 7.33 (d, J = 1.6Hz, 1H), 3.65 (d, J = 10.8 Hz, 2H), 3.59 (d, J = 11.6 Hz, 2H), 3.10 (t, J = 12.0Hz, 2H), 3.06-2.85 (m, 4H), 2.14-1.99 (m, 2H), 1.97-1.87 (m, 2H), 1.84-1.72(m, 4H), 1.70-1.57 (m, 2H), 1.21-1.11 (m, 1H).

[0080] ESI-MS: m / z = 492 [M+H] + .

[0081] Embodiment 2: N -((1-(2-(4-cyanophenyl)-5-trifluoromethyl-7-azaindol-4-yl)azetidin-3-yl)methyl)cyclobutylcarboxamide (B002)

[0082]

[0083] Referring to step 1 of Example 1, intermediate 2-2 can be obtained by replacing 1-1 with intermediate 2-1. ESI-MS: m / z = 269[M+H] + .

[0084] Referring to step 2 of Example 1, intermediate 2-2 was used instead of 1-2 to obtain intermediate 2-3. ESI-MS: m / z = 169[M+H] + .

[0085] Step 1: Synthesis of intermediate 2-5

[0086] 2-4 (1.62 g, 10 mmol), silver trifluoroacetate (2.21 g, 10 mmol) and iodine (2.53 g, 10 mmol) were dissolved in 40 mL of dichloroethane at 90 °C. o After reacting for 7 hours at 40 °C, the mixture was cooled to room temperature, iodine (2.53 g, 10 mmol) was added, and the temperature was raised to 90 °C. o C and continue to react for 16 hours. After the reaction is complete, cool to room temperature, filter, add 50 mL of water to dilute the filtrate, extract with dichloromethane (60 mL × 3), combine the organic phases, dry over anhydrous sodium sulfate, filter, and spin dry. The crude product is purified by column chromatography (PE:EA = 50:1-20:1-10:1) to obtain 1.90 g of a light red solid (2-5). ESI-MS: m / z=289 [M+H] + .

[0087] Step 2: Synthesis of intermediate 2-6

[0088] 2-5 (1.90 g, 6.60 mmol), 4-ethynylbenzonitrile (0.92 g, 7.26 mmol), cuprous iodide (62.7 mg, 0.330 mmol), bis(triphenylphosphine)palladium dichloride (0.23 g, 0.330 mmol) and diisopropylamine (2.00 g, 19.8 mmol) were dissolved in 25 mL DMF. o C for 5 hours. After the reaction is complete, cool to room temperature, add 50 mL of water to dilute the reaction solution, extract with ethyl acetate (50 mL× 3), combine the organic phases, dry over anhydrous sodium sulfate, filter, and spin dry. The crude product is purified by column chromatography (PE:EA = 10:1~5:1~3:1) to obtain 2.04 g of white solid (2-6). ESI-MS: m / z =288 [M+H] + .

[0089] Step 3: Synthesis of intermediate 2-7

[0090] Sodium hydride (0.569 g, 14.2 mmol, 60%) w ) was dissolved in 25 mL of anhydrous NMP, 0 o 2-6 (2.04 g, 7.10 mmol) was added at 80 °C and then the temperature was raised to 80o C for 2.5 hours. After the reaction is complete, cool to room temperature, add 50 mL of water to dilute the reaction solution, extract with ethyl acetate (50 mL× 3), combine the organic phases, dry over anhydrous sodium sulfate, filter, and spin dry. The crude product is purified by column chromatography (PE:EA = 10:1~5:1~4:1) to obtain 1.41 g of white solid (2-7). ESI-MS: m / z = 288 [M+H] + .

[0091] Step 4: Synthesis of intermediate 2-8

[0092] Dissolve 2-7 (1.41 g, 4.90 mmol) and m-chloroperbenzoic acid (0.932 g, 5.40 mmol) in 25 mL of tetrahydrofuran. o After the reaction was complete, the reaction solution was dried by spin drying, slurried with a DCM:PE = 1:2 mixed solvent, filtered and vacuum dried, and then dissolved in 30 mL of phosphorus oxychloride and 85 o C for 4 hours. After the reaction is complete, cool to room temperature, slowly add the reaction solution dropwise to ice water to quench, extract with ethyl acetate (70 mL × 3), combine the organic phases, dry over anhydrous sodium sulfate, filter, and spin dry. The crude product is purified by column chromatography (PE:EA = 10:1~6:1) to obtain 800.0 mg (2-8) of white solid. ESI-MS: m / z = 322 [M+H] + .

[0093] Referring to step 3 of Example 1, intermediate 2-8 was used to replace 1-4 to obtain intermediate 2-9. ESI-MS: m / z = 452[M+H] + .

[0094] Step 5: Synthesis of Intermediate 2-10

[0095] 2-9 (226.0 mg, 0.5 mmol), 2-3 (168.0 mg, 1 mmol), cesium carbonate (326.0 mg, 1 mmol), palladium acetate (12.0 mg, 0.05 mmol) and XantPhos (28.9 mg, 0.05 mmol) were dissolved in 10 mL of dioxane, and the atmosphere was replaced with nitrogen at 110 °C. oC for 16 hours. After the reaction is complete, cool to room temperature, add 30 mL of water to dilute the reaction solution, extract with ethyl acetate (50 mL × 3), combine the organic phases, dry over anhydrous sodium sulfate, filter, and spin dry. The crude product is purified by column chromatography (PE:EA = 5:1~3:1~2:1) to obtain 30.0 mg of light blue solid (2-10). ESI-MS: m / z = 584 [M+H] + .

[0096] Referring to step 8 of Example 1, intermediate 2-10 was used instead of 1-8 to obtain B002, 10.0 mg.

[0097] 1 H NMR (400 MHz, DMSO- d 6) δ 12.38 (s, 1H), 8.14 (s, 1H), 8.09 (d, J =8.4 Hz, 2H), 7.92 (t, J = 6.0 Hz, 1H), 7.89 (d, J = 8.0 Hz, 2H), 7.35 (s, 1H), 4.61 (t, J = 8.8 Hz, 2H), 4.27 (t, J = 7.2 Hz, 2H), 3.05-2.92 (m, 1H), 2.90-2.78(m, 1H), 2.15-2.04 (m, 2H), 2.03-1.93 (m, 2H), 1.91-1.79 (m, 1H), 1.79-1.67(m, 1H).

[0098] ESI-MS: m / z = 454 [M+H] + .

[0099] Embodiment 3: N -(3-((2-(4-cyanophenyl)-5-trifluoromethyl-7-azaindol-4-yl)amino)propyl)cyclobutylcarboxamide (B003)

[0100]

[0101] Referring to step 1 of Example 1, intermediate 3-1 was used instead of 1-1 to obtain intermediate 3-2. ESI-MS: m / z = 257[M+H] + .

[0102] Referring to step 2 of Example 1, intermediate 3-2 was used to replace 1-2 to obtain intermediate 3-3. ESI-MS: m / z = 157[M+H] + .

[0103] Referring to step 5 of Example 2, intermediate 3-3 was used instead of 2-3 to obtain intermediate 3-4. ESI-MS: m / z = 572[M+H] + .

[0104] Referring to step 8 of Example 1, intermediate 3-4 was used instead of 1-8 to obtain B003, 80.0 mg.

[0105] 1 H NMR (400 MHz, DMSO- d 6) δ 8.22 (s, 1H), 8.14 (d, J = 8.4 Hz, 2H),7.93 (d, J = 8.4 Hz, 2H), 7.81 (t, J = 6.0 Hz, 2H), 7.46 (s, 1H), 7.04 (s, 1H), 3.75 (q, J = 6.4 Hz, 2H), 3.17 (q, J = 6.4 Hz, 2H), 2.98 (p, J = 8.4 Hz, 1H), 2.17-2.04 (m, 2H), 2.03-1.91 (m, 2H), 1.91-1.66 (m, 4H).

[0106] ESI-MS: m / z = 442 [M+H] + .

[0107] Embodiment 4: N -(3-((2-(4-cyanophenyl)-5-ethynyl-7-azaindol-4-yl)amino)propyl)cyclobutylcarboxamide (B004)

[0108]

[0109] Referring to step 2 of Example 2, intermediate 4-1 was used instead of 2-5 to obtain intermediate 4-2. ESI-MS: m / z = 298[M+H] + .

[0110] Referring to step 3 of Example 2, intermediate 4-2 was used instead of 2-6 to obtain intermediate 4-3. ESI-MS: m / z = 298[M+H] +.

[0111] Referring to step 2 of Example 2, intermediate 4-3 is used instead of 2-5, trimethylsilyl acetylene is used instead of 4-ethynyl benzonitrile, 90 o C for 17 hours to obtain intermediate 4-4. ESI-MS: m / z = 316 [M+H] + .

[0112] Referring to step 4 of Example 2, intermediate 4-4 was used instead of 2-7 to obtain intermediate 4-5. ESI-MS: m / z = 350[M+H] + .

[0113] Referring to step 3 of Example 1, intermediate 4-5 was used to replace 1-4 to obtain intermediate 4-6. ESI-MS: m / z = 480[M+H] + .

[0114] Referring to step 6 of Example 1, intermediate 4-6 is used to replace 1-7, and intermediate 3-3 is used to replace 1-3 to obtain intermediate 4-7. ESI-MS: m / z = 600 [M+H] + .

[0115] Referring to step 8 of Example 1, intermediate 4-7 was used instead of 1-8 to obtain B004, 10.0 mg.

[0116] 1 H NMR (400 MHz, DMSO- d 6) δ 12.20 (s, 1H), 8.09 (d, J = 8.0 Hz, 2H),7.96 (s, 1H), 7.88 (d, J = 8.0 Hz, 2H), 7.78 (t, J = 6.0 Hz, 1H), 7.31 (s, 1H), 6.39 (t, J = 6.4 Hz, 1H), 4.38 (s, 1H), 3.69 (q, J = 6.8 Hz, 2H), 3.17 (q, J = 6.5Hz, 2H), 2.98 (p, J = 8.5 Hz, 1H), 2.17-2.05 (m, 2H), 2.01-1.92 (m, 2H), 1.90-1.65 (m, 4H).

[0117] ESI-MS: m / z = 398 [M+H] +.

[0118] Example 5: 4-(4-((3-(piperidin-2-one-1-yl)propyl)amino)-5-(trifluoromethyl)-7-azaindol-2-yl)benzonitrile (B005)

[0119]

[0120] Step 1: Synthesis of intermediate 5-2

[0121] Dissolve 5-1 (8.0 g, 36.8 mmol) in 80 mL of acetonitrile, add cesium carbonate (24.9 g, 73.6 mmol) and o C for 10 minutes, 1,3-dibromopropane (14.9 g, 73.6 mmol) was added dropwise, and 25 o C for 17 hours. After the reaction is complete, filter and spin dry the filtrate. The crude product is purified by column chromatography (PE:EA = 100:0~20:1) to obtain 8.9 g (5-2) of colorless liquid. ESI-MS: m / z = 338 [M+H] + .

[0122] Step 2: Synthesis of intermediate 5-3

[0123] 2-Piperidone (2.1 g, 20.5 mmol) was added to 25 mL DMF and stirred at 0. o NaH (902 mg, 22.6 mmol) was slowly added in portions at 0. o C for 15 minutes, add 5-2 (8.3 g, 24.6 mmol), 50 o C for 17 hours. After the reaction is complete, add 50 mL of water to the reaction system, extract with ethyl acetate (50 mL × 5), combine the organic phases, wash with water (50 mL × 4), wash with saturated brine (60 mL × 2), dry with anhydrous sodium sulfate, filter, and spin dry. The crude product was purified by column chromatography (PE:EA = 100:0~0:100) to obtain 2.8 g (5-3) of light yellow liquid with a yield of 38%. ESI-MS: m / z =357 [M+H] + .

[0124] Step 3: Synthesis of intermediate 5-4

[0125] To the reaction bottle containing 5-3 (2.8 g, 7.86 mmol), 30 mL of hydrochloric acid-ethyl acetate solution (2M) was added. oC for 3 hours. When the reaction is complete, the solvent is directly dried to obtain 3.0 g of white solid (5-4 hydrochloride). ESI-MS: m / z = 157 [M+H] + .

[0126] Referring to step 5 of Example 2, intermediate 5-5 can be obtained by replacing 2-3 with intermediate 5-4. ESI-MS: m / z = 572[M+H] + .

[0127] Referring to step 8 of Example 1, B005 can be obtained by replacing 1-8 with intermediate 5-5. ESI-MS: m / z = 442 [M+H] + .

[0128] Example 6: 4-(4-((3-(pyridin-2-one-1-yl)propyl)amino)-5-(trifluoromethyl)-7-azaindol-2-yl)benzonitrile (B006)

[0129]

[0130] Referring to steps 2 and 3 of Example 5, 2-pyridone was used instead of 2-piperidone to obtain the hydrochloride salt of intermediate 6-2. ESI-MS: m / z = 153 [M+H] + .

[0131] Referring to step 5 of Example 2, intermediate 6-2 was used instead of 2-3 to obtain intermediate 6-3. ESI-MS: m / z = 568[M+H] + .

[0132] Referring to step 8 of Example 1, B006 can be obtained by replacing 1-8 with intermediate 6-3. ESI-MS: m / z = 438 [M+H] + .

[0133] Example 7: 4-(4-((3-(tetrahydropyrimidin-2-one-1-yl)propyl)amino)-5-(trifluoromethyl)-7-azaindol-2-yl)benzonitrile (B007)

[0134]

[0135] Step 1: Synthesis of intermediate 7-3

[0136] Place intermediate 7-1 (1.3 g, 10.0 mmol) and dimethyl carbonate (0.9 g, 10.0 mmol) in a single-necked bottle, and add 7-2 (70.0 mg, 0.5 mmol) dropwise. oC and reflux for 1 hour. After the reaction is complete, 1.6 g of a colorless oily crude product (7-3) is obtained. ESI-MS: m / z = 158 [M+H] + .

[0137] Referring to step 5 of Example 2, intermediate 7-3 was used instead of 2-3 to obtain intermediate 7-4. ESI-MS: m / z = 573[M+H] + .

[0138] Referring to step 8 of Example 1, B007 can be obtained by replacing 1-8 with intermediate 7-4. ESI-MS: m / z = 443 [M+H] + .

[0139] Example 8: 4-(4-((3-(1,4-oxoazepan-5-one-4-yl)propyl)amino)-5-(trifluoromethyl)-7-azaindol-2-yl)benzonitrile (B008)

[0140]

[0141] Referring to steps 2 and 3 of Example 5, the hydrochloride of intermediate 8-3 can be obtained by replacing 2-piperidone with 8-1. ESI-MS: m / z = 173 [M+H] + .

[0142] Referring to step 5 of Example 2, intermediate 8-4 can be obtained by replacing 2-3 with intermediate 8-3. ESI-MS: m / z = 588[M+H] + .

[0143] Referring to step 8 of Example 1, B008 can be obtained by replacing 1-8 with intermediate 8-4. ESI-MS: m / z = 458 [M+H] + .

[0144] Embodiment 9: N -(3-((2-(4-cyanophenyl)-5-(trifluoromethyl)-7-azaindol-4-yl)amino)-2-methylpropyl)cyclobutylcarboxamide (B009)

[0145]

[0146] Referring to steps 1 and 2 of Example 1, the hydrochloride of intermediate 9-3 can be obtained by replacing 1-1 with intermediate 9-1. ESI-MS: m / z = 171 [M+H] + .

[0147] Referring to step 5 of Example 2, intermediate 9-4 can be obtained by replacing 2-3 with intermediate 9-3. ESI-MS: m / z = 586[M+H] + .

[0148] Referring to step 8 of Example 1, B009 can be obtained by replacing 1-8 with intermediate 9-4.

[0149] 1 H NMR (400 MHz, DMSO- d 6) δ 12.40 (s, 1H), 8.13 (s, 1H), 8.12 (d, J =8.4 Hz, 2H), 8.11 (s, 1H), 7.90 (d, J = 8.0 Hz, 2H), 7.81 (t, J = 6.0 Hz, 1H),7.31 (s, 1H), 6.54 (d, J = 6.4 Hz, 1H), 3.59 (dt, J = 14.0, 6.4 Hz, 1H), 3.48(dt, J = 14.4, 6.8 Hz, 1H), 3.19-3.09 (m, 1H), 3.09-2.95 (m, 2H), 2.19-2.06 (m,2H), 2.06-1.92 (m, 3H), 1.92-1.78 (m, 1H), 1.77-1.66 (m, 1H), 0.91 (d, J = 6.4 Hz, 3H).

[0150] ESI-MS: m / z = 456 [M+H] + .

[0151] Embodiment 10: N -(3-((2-(4-cyanophenyl)-5-(trifluoromethyl)-7-azaindol-4-yl)amino)-2,2-dimethylpropyl)cyclobutylcarboxamide (B010)

[0152]

[0153] Step 1: Synthesis of intermediate 10-2

[0154] 10-1 (1.5 g, 14.7 mmol) was dissolved in 15.0 mL of dichloromethane. oC, add triethylamine (2.97 g, 29.4 mmol), stir for 10 min, add Boc2O (3.2 g, 14.7 mmol) dissolved in 5 mL of dichloromethane, and o C for 4 hours. After the reaction is complete, filter and spin dry the filtrate. The crude product is purified by column chromatography (EA:MeOH = 100:0~10:1) to obtain 940.0 mg (10-2) of white solid. ESI-MS: m / z = 203 [M+H] + .

[0155] Referring to Example 9, B010 can be obtained by replacing 9-1 with intermediate 10-2. ESI-MS: m / z = 470 [M+H] + .

[0156] Embodiment 11: N -(3-((2-(4-cyanobenzyl)-5-(trifluoromethyl)-7-azaindol-4-yl)amino)propyl)cyclobutylcarboxamide (B011)

[0157]

[0158] Step 1: Synthesis of Intermediate 11-1

[0159] Dissolve 1-4 (15.2 g, 100.0 mmol) in 300.0 mL of anhydrous tetrahydrofuran, and add sodium hydride (5.6 g, 140.0 mmol, 60%) in portions under an ice-water bath. w ), after stirring for 20 minutes, triisopropylsilyl chloride (30.9 g, 160.0 mmol) was slowly added, and the mixture was returned to room temperature and stirred overnight. After the reaction was complete, the mixture was spin-dried. The crude product was purified by column chromatography (PE:EA = 100:0~10:1) to obtain 28.0 g of solid (11-1). ESI-MS: m / z = 309 [M+H] + .

[0160] Step 2: Synthesis of Intermediate 11-2

[0161] 11-1 (28.0 g, 90.9 mmol) was dissolved in 250.0 mL of anhydrous tetrahydrofuran and heated at -80 o140.0 mL of sec-butyl lithium (181.8 mmol, 1.3 M in n-hexane) was slowly added dropwise at 40 °C. After stirring for 2 hours, iodine (34.6 g, 136.4 mmol) was dissolved in 100.0 mL of tetrahydrofuran and added dropwise to the reaction solution. The reaction was continued for 2 hours. After the reaction was complete, 50.0 mL of saturated ammonium chloride solution was added to quench the reaction. After returning to room temperature, the mixture was extracted with ethyl acetate (200 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and dried. The crude product was purified by column chromatography (PE:EA = 100:0~10:1) to obtain 20.0 g of solid (11-2). ESI-MS: m / z =435 [M+H] + .

[0162] Step 3: Synthesis of Intermediate 11-3

[0163] Dissolve 11-2 (20.0 g, 46.0 mmol) in 100.0 mL of anhydrous tetrahydrofuran, add 46.0 mL of tetrabutylammonium fluoride in tetrahydrofuran solution (1 M) dropwise under an ice-water bath, and stir for 1 hour. When the reaction is complete, add 100 mL of water to quench the reaction, extract with ethyl acetate (100 mL×3), combine the organic phases, dry over anhydrous sodium sulfate, filter, and spin dry. The crude product is purified by column chromatography (PE:EA = 10:0~2:1) to obtain 10.0 g of solid (11-3). ESI-MS: m / z = 279 [M+H] + .

[0164] Referring to step 3 of Example 1, intermediate 11-3 was used to replace 1-4 to obtain intermediate 11-4. ESI-MS: m / z =409 [M+H] + .

[0165] Step 4: Synthesis of Intermediate 11-5

[0166] 11-4 (2.5 g, 6.1 mmol), methyl 2,2-difluoro-2-(fluorosulfonyl)acetate (3.5 g, 18.4 mmol), and cuprous iodide (1.2 g, 6.1 mmol) were dissolved in 6 mL DMF. o C for 2 hours. When the reaction is complete, add 30 mL of water to quench the reaction, extract with ethyl acetate and saturated water (50 mL×3), combine the organic phases, dry over anhydrous sodium sulfate, filter, and spin dry. The crude product is purified by column chromatography (PE:EA=100:1~10:1) to obtain 1.3 g of solid (11-5). ESI-MS: m / z =351 [M+H] + .

[0167] Referring to step 4 of Example 1, intermediate 11-5 was used to replace 1-5 to obtain intermediate 11-6. ESI-MS: m / z =395 [M+H] + .

[0168] Step 5: Synthesis of Intermediate 11-7

[0169] Dissolve 11-6 (158.0 mg, 0.40 mmol), p-cyanobenzyl bromide (80.0 mg, 0.40 mmol), Pd(PPh3)4 (46.0 mg, 0.04 mmol) and sodium carbonate (84.0 mg, 0.80 mmol) in 1 mL of dioxane, replace nitrogen atmosphere, and heat at 85 °C. o C for 6 hours. When the reaction is complete, add 10 mL of saturated brine to dilute the reaction solution, extract with ethyl acetate (10 mL × 3), combine the organic phases, dry over anhydrous sodium sulfate, filter, and spin dry. The crude product is purified by column chromatography (PE:EA = 50:1~10:1) to obtain 65.0 mg of the product solid (11-7). ESI-MS: m / z = 466 [M+H] + .

[0170] Step 6: Synthesis of Intermediate 11-8

[0171] 11-7 (60.0 mg, 0.13 mmol) was dissolved in 0.5 mL 1,3-propylenediamine and heated in a microwave oven at 120 o C for 1 hour. When the reaction is complete, add 10 mL of saturated brine to dilute the reaction solution, extract with ethyl acetate (10 mL × 3), combine the organic phases, dry over anhydrous sodium sulfate, filter, and spin dry. The crude product is purified by column chromatography (DCM:MeOH = 20:1) to obtain 40.0 mg of solid (11-8). ESI-MS: m / z = 504 [M+H] + .

[0172] Referring to step 1 of Example 1, intermediate 11-8 was used instead of 1-1 to obtain intermediate 11-9. ESI-MS: m / z =586 [M+H] + .

[0173] Referring to step 8 of Example 1, intermediate 11-9 was used instead of 1-8 to obtain B011, 5.0 mg.

[0174] 1 H NMR (400 MHz, DMSO- d 6) δ 11.63 (s, 1H), 7.94 (s, 1H), 7.74 (d, J=8.4 Hz, 2H), 7.70 (t, J = 6.0 Hz, 1H), 7.45 (d, J = 8.4 Hz, 2H), 6.44 (s, 1H), 6.18 (t, J = 6.0 Hz, 1H), 4.07 (s, 2H), 3.52 (q, J = 6.4 Hz, 2H), 3.07 (q, J = 6.4Hz, 2H), 2.93 (p, J = 8.0 Hz, 1H), 2.13-2.01 (m, 2H), 2.00-1.90 (m, 2H), 1.87-1.74 (m, 1H), 1.72-1.61 (m, 3H).

[0175] ESI-MS: m / z = 456 [M+H] + .

[0176] Embodiment 12: N -(3-((2-(3-(4-methylpiperazin-1-yl)phenyl)-5-(trifluoromethyl)-7-azaindol-4-yl)amino)propyl)cyclobutylcarboxamide (B012)

[0177]

[0178] Step 1: Synthesis of intermediate 12-1

[0179] 1,3-Dibromobenzene (1.18 g, 5.0 mmol), N-methylpiperazine (0.50 g, 5.0 mmol), Pd2(dba)3 (0.23 g, 0.25 mmol), XantPhos (0.29 g, 0.5 mmol) and potassium tert-butoxide (1.73 g, 15.0 mmol) were dissolved in toluene and the atmosphere was replaced with nitrogen. o C for 8 hours. After the reaction is complete, spin dry. The crude product is purified by column chromatography (PE:EA = 20 / 1) to obtain 0.84 g of the intermediate (12-1). ESI-MS: m / z = 255 [M+H] + .

[0180] Referring to steps 5 and 6 of Example 11, intermediate 12-1 was used to replace p-cyanobenzyl bromide to obtain intermediate 12-3. ESI-MS: m / z = 563 [M+H] + .

[0181] Referring to step 1 of Example 1, intermediate 12-3 was used instead of 1-1 to obtain intermediate 12-4. ESI-MS: m / z = 645 [M+H] + .

[0182] Referring to step 8 of Example 1, intermediate 12-4 was used instead of 1-8 to obtain B012, 5.0 mg.

[0183] 1 H NMR (400 MHz, DMSO- d 6) δ 12.08 (s, 1H), 8.02 (s, 1H), 7.75 (t, J =6.0 Hz, 1H), 7.44 (s, 1H), 7.31 (d, J = 7.6 Hz, 1H), 7.22 (t, J = 8.0 Hz, 1H),7.05 (s, 1H), 6.83 (dd, J = 8.0, 2.4 Hz, 1H), 6.25 (t, J = 6.4 Hz, 1H), 3.65 (q, J = 6.4 Hz, 2H), 3.22 (t, J = 4.8 Hz, 4H), 3.12 (q, J = 6.4 Hz, 2H), 2.94 (p, J = 8.0Hz, 1H), 2.51 (t, J = 4.8 Hz, 4H), 2.24 (s, 3H), 2.13-2.01 (m, 2H), 1.98-1.88 (m, 2H), 1.86-1.78 (m, 1H), 1.76-1.66 (m, 3H).

[0184] ESI-MS: m / z = 515 [M+H] + .

[0185] Example 13: S )- N -(2-Fluoro-3-((2-(3-(4-methylpiperazin-1-yl)phenyl)-5-(trifluoromethyl)-7-azaindol-4-yl)amino)propyl)cyclobutylcarboxamide (B013)

[0186]

[0187] Step 1: Synthesis of intermediate 13-2

[0188] Dissolve 13-1 (4.0 g, 19.70 mmol) in 28.0 mL of ammonia methanol solution (7 M) and react overnight at room temperature. After the reaction is complete, spin dry to obtain 3.5 g of white solid (13-2). ESI-MS: m / z = 221 [M+H] + .

[0189] Referring to step 6 of Example 11, intermediate 13-2 was used to replace 1,3-propylenediamine to obtain intermediate 13-3. ESI-MS: m / z = 709 [M+H] + .

[0190] Step 2: Synthesis of Intermediate 13-4

[0191] Dissolve 13-3 (1.4 g, 2.0 mmol) in 20 mL of ethanol, add hydrazine hydrate (0.76 g, 6.0 mmol) dropwise, and heat to 70 o C, react for 6 hours. After the reaction is complete, filter and spin dry the filtrate to obtain 0.87 g of crude product (13-4), which is directly used in the next step. ESI-MS: m / z = 579 [M+H] + .

[0192] Referring to step 1 of Example 1, intermediate 13-4 was used instead of 1-1 to obtain intermediate 13-5. ESI-MS: m / z = 661 [M+H] + .

[0193] Step 3: Synthesis of Intermediate 13-6

[0194] 13-5 (0.66 g, 1.0 mmol) was dissolved in 5 mL of dichloromethane, pyridine (162.5 μL, 2.0 mmol) was added, and DAST (265.0 μL, 2.0 mmol) was added dropwise under ice-water bath. o C for 7 days. The raw material was not completely reacted and was dried by spin drying. The crude product was purified by column chromatography (PE:EA = 2:1) to obtain 65.3 mg of the intermediate (13-6). ESI-MS: m / z = 663 [M+H] + .

[0195] Referring to step 8 of Example 1, B013 can be obtained by replacing 1-8 with intermediate 13-6. ESI-MS: m / z = 533 [M+H] + .

[0196] Embodiment 14: N -(3-((2-(3-((3 S ,5R )-4-ethyl-3,5-dimethylpiperazin-1-yl)phenyl)-5-(trifluoromethyl)-7-azaindol-4-yl)amino)propyl)cyclobutylcarboxamide (B014)

[0197]

[0198] Step 1: Synthesis of intermediate 14-2

[0199] 14-1 (500.0 mg, 2.33 mmol) was dissolved in 10 mL of tetrahydrofuran, and NaH (280.0 mg, 7.0 mmol, 60% w ), stirred at room temperature for 30 minutes, added iodoethane (730.0 mg, 4.67 mmol), 80 o C, the reaction was continued for 7 hours, ethyl iodide (730.0 mg, 4.67 mmol) was added, and the mixture was continued at 80 o C for 16 hours. After the reaction is complete, the reaction system is slowly dripped into 20 ml of water to quench, extracted with ethyl acetate (25 mL × 4), the organic phases are combined, washed with saturated brine (40 mL × 2), dried over anhydrous sodium sulfate, filtered, and spin-dried. The crude product is purified by column chromatography (PE:EA = 100:0~0:1) to obtain 473.0 mg (14-2) of yellow liquid. ESI-MS: m / z = 243 [M+H] + .

[0200] Referring to step 2 of Example 1, the hydrochloride of intermediate 14-3 can be obtained by replacing 1-2 with intermediate 14-2. ESI-MS: m / z = 143 [M+H] + .

[0201] Referring to Example 12, intermediate 14-3 was used to replace N-methylpiperazine to obtain B014. ESI-MS: m / z = 557[M+H] + .

[0202] Embodiment 15: N -(3-((2-(3-(8-ethyl-3,8-diazabicyclo[3.2.1]oct-3-yl)phenyl)-5-(trifluoromethyl)-7-azaindol-4-yl)amino)propyl)cyclobutylcarboxamide (B015)

[0203]

[0204] Referring to Example 14, B015 can be obtained by replacing 14-1 with intermediate 15-1. ESI-MS: m / z = 555 [M+H] + .

[0205] Example 16: R )-N-(3-((2-(3-(hexahydropyrrolo[1,2-a]pyrazine-2( 1H )-(4-(trifluoromethyl)-7-azaindol-4-yl)amino)propyl)cyclobutylcarboxamide (B016)

[0206]

[0207] Referring to Example 12, intermediate 16-1 was used to replace N-methylpiperazine to obtain B016. ESI-MS: m / z = 541[M+H] + .

[0208] Embodiment 17: N -(3-((2-(3-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)phenyl)-5-(trifluoromethyl)-7-azaindol-4-yl)amino)propyl)cyclobutylcarboxamide (B017)

[0209]

[0210] Referring to step 5 of Example 11, intermediate 17-1 was used instead of 11-6, and 1,3-dibromobenzene was used instead of p-cyanobenzyl bromide to obtain intermediate 17-2. ESI-MS: m / z = 252 [M+H] + .

[0211] Referring to Example 12, B017 can be obtained by replacing 12-1 with intermediate 17-2. ESI-MS: m / z = 512 [M+H] + .

[0212] Embodiment 18: N -(3-((2-(3-(1-methylpiperidin-4-yl)phenyl)-5-(trifluoromethyl)-7-azaindol-4-yl)amino)propyl)cyclobutylcarboxamide (B018)

[0213]

[0214] B017 (51.0 mg, 0.1 mmol) was dissolved in 0.5 mL methanol, 10%w palladium carbon (10.0 mg) was added, hydrogen was replaced, and the reaction was carried out at 45°C for 2 hours. After the reaction was complete, it was filtered with diatomaceous earth, washed with methanol three times, the organic phases were combined, dried by spin drying, and the crude product was purified by column chromatography to obtain B018.

[0215] ESI-MS: m / z =514 [M+H] + .

[0216] Effect experiment: In vitro ULKs kinase activity inhibition experiment

[0217] Experimental purpose: The ADP-GLO method was used to detect the in vitro inhibitory activity of the test compounds (see Table 1) on the ULK1 and ULK2 kinase activities.

[0218] Experimental methods:

[0219] Compounds were dissolved in 100% DMSO to prepare 10 mM stock solutions. Kinases, substrates, ATP, and compounds were diluted in kinase reaction buffer (50 mM HEPES, 10 mM MgCl2, 0.01% Brij97, 1 mM EGTA, 2 mM DTT, 0.1 mg / mL BSA). 1 μL of compound was incubated with 1.5 μL ULK1 enzyme or 3 μL ULK2 enzyme at room temperature for 10 min, 10 μM ATP and 100 μg / mL MBP were added to prepare a 50 μL total reaction system, centrifuged at 1000 rpm for 10 s, and incubated at room temperature for 60 min. Then 5 μL of the reaction solution was taken, 5 μL ADP-Glo™ reagent was added, centrifuged at 1000 rpm for 10 s, incubated at room temperature for 40 min, and 10 μL kinase detection reagent was added. The reaction was centrifuged at 1000 rpm for 10 s, incubated at room temperature for 30 min, and luminescence was detected.

[0220] Data Analysis:

[0221] The log value of the concentration was used as the X-axis, and the percentage inhibition rate was used as the Y-axis. The [Inhibitor] vs. normalized response -- Variable slope of the analysis software GraphPad Prism9 was used to fit the dose-effect curve to obtain the IC value of each compound on the enzyme activity. 50 The test results are shown in Table 1 below.

[0222] Table 1

[0223]

[0224] Note: NT means the data has not been tested.

[0225] According to the test results in Table 1 above, it can be seen that the compounds prepared by the present invention can effectively inhibit the activity of ULKs kinases, especially the activities of ULK1 kinase and ULK2 kinase.

[0226] Finally, it should be pointed out that the above content is only used to illustrate the technical solution of the present invention and does not limit the protection scope of the present invention. For ordinary technicians in this field, simple modifications or equivalent substitutions do not violate the essence and scope of the technical solution of the present invention.

Claims

1. A 4-amino-7-azaindole compound or a pharmaceutically acceptable salt thereof, characterized in that: Its structural formula is shown in the following formula (3): （3） in, R 4 Selected from hydrogen or C 1-3 The alkyl group; R 6 is selected from trifluoromethyl or ethynyl; R 7 and R 8 At least one of them is hydrogen and the other one is selected from , , , or cyano.

2. The 4-amino-7-azaindole compound or pharmaceutically acceptable salt according to claim 1, characterized in that: The 4-amino-7-azaindole compound or pharmaceutically acceptable salt is one of the following compounds: 。 3. A pharmaceutical composition, characterized in that: The pharmaceutical composition comprises the 4-amino-7-azaindole compound or pharmaceutically acceptable salt according to claim 1 or 2, and a pharmaceutically acceptable carrier.

4. Use of the 4-amino-7-azaindole compound or the pharmaceutically acceptable salt according to claim 1 or 2 or the pharmaceutical composition according to claim 3 in the preparation of a drug for treating diseases caused by abnormal activation of ULK kinase.

5. The use according to claim 4, characterized in that: The disease is an autoimmune disease, cardiovascular disease, metabolic disease, liver disease, neurodegenerative disease, infectious disease or cancer; and the cancer is metastatic cancer.

6. The use according to claim 5, characterized in that: The metastatic cancer is a gastrointestinal stromal tumor, esophageal cancer, gastric cancer, melanoma, glioma, glioblastoma, ovarian cancer, bladder cancer, pancreatic cancer, prostate cancer, lung cancer, breast cancer, kidney cancer, liver cancer, osteosarcoma, multiple myeloma, cervical cancer, bone metastasis, papillary thyroid cancer, non-small cell lung cancer or colorectal cancer.

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