Tricyclic BCL6 degradation agent and use thereof

Abstract

Disclosed in the present invention are a tricyclic BCL6 degradation agent and the use thereof. The tricyclic compound provided by the present invention or a pharmaceutically acceptable salt and solvate thereof has a novel structure and exhibits an excellent degradation effect on BCL6 protein. Therefore, the compound provided by the present invention or a composition containing the compound provided by the present invention exhibits a promising potential for development as drugs for treating diseases that can be treated or relieved by means of degrading the BCL6 protein, e.g., cancers such as Hodgkin lymphoma, B-cell non-Hodgkin lymphoma, T-cell non-Hodgkin lymphoma, NK / T cell non-Hodgkin lymphoma, or diffuse large B-cell lymphoma.

Description

Tricyclic bcl6 degraders and uses thereof TECHNICAL FIELD

[0001] The present application belongs to the field of pharmaceutical chemistry, and relates to synthesis of new compounds and medical uses thereof, in particular to tricyclic BCL6 degraders and uses thereof. BACKGROUND

[0002] B-cell lymphoma 6 (BCL6) is an important transcriptional repressor in humoral immune response. Human BCL6 gene is about 24 kb, encoding a BCL6 protein of about 95 kD. BCL6 protein is a member of the POZ / BTB / Zinc finger protein family, which is mainly composed of three parts: 1) the POZ / BTB domain at the amino terminal end, which is the main functional region for transcriptional repression. BCL6 needs to form a BTB dimer to exert transcriptional repression. Three main transcriptional co-repressors SMRT, BCOR or NCOR bind to the BTB binding site in a competitive manner to exert transcriptional repression together, and participate in the transcriptional regulation in the early stage of germinal center (GC) formation. 2) the middle region (three PEST domains, also known as RD2 domains), which mainly recruits auxiliary factors such as MTA3 or CTBP1 to stabilize proteins. 3) the zinc finger domain at the carboxy terminal end, which is composed of six identical zinc finger structures and mainly plays a role in binding DNA, which is a prerequisite for BCL6 to exert transcriptional repression.

[0003] BCL6 is a transcriptional repressor that is essential for GC development and maintenance. In the GC response, chromosomal translocation and point mutation of BCL6 lead to persistent high expression of BCL6 protein, which promotes malignant proliferation of B cells and further leads to the occurrence of B-cell lymphoma. Most non-Hodgkin lymphoma (NHL) is derived from GC, among which diffuse large B-cell lymphoma (DLBCL) is the most common subtype. BCL6 is recognized as an oncogenic driver of DLBCL. Many preclinical studies have also shown that blocking the interaction between BCL6-BTB domain and its transcriptional co-repressor can inhibit GC formation and DLBCL cell proliferation, which is an effective strategy for treating DLBCL, and has good safety without causing toxic side effects and inflammatory reactions driven by macrophages.

[0004] BCL6 is a highly potential cancer treatment target, including but not limited to Hodgkin's lymphoma, B-cell derived non-Hodgkin's lymphoma, T-cell derived non-Hodgkin's lymphoma, NK / T-cell derived non-Hodgkin's lymphoma or diffuse large B-cell lymphoma. The currently reported BCL6 small molecule inhibitors have some defects, such as poor target binding activity, insufficient in vitro and in vivo cell activity, unclear mechanism, poor drug-like properties, etc. The currently reported BCL6 small molecule degraders have a clear mechanism, which induce the aggregation of BCL6 protein in cells and specifically degrade BCL6. The unique mechanism makes the anti-proliferation effect of BI-3802 (BCL6 small molecule degrader) comparable to that of CRISPR-Cas9 knockout BCL6 gene. There are about 18 BCL6 drugs under research, covering small molecule inhibitors (8 types), PROTAC (5 types) and molecular glue (5 types), and only the PROTAC drug (BMS-986458) of BMS has entered phase I clinical study. Therefore, it is still urgent and necessary to develop a small molecule degrader with novel structure, good drug property and safety, and capable of rapidly and efficiently degrading BCL6. SUMMARY

[0005] The purpose of the present application is to overcome the deficiencies of the prior art and provide a tricyclic BCL6 degrader and uses thereof.

[0006] The above-mentioned purposes of the present application are achieved by the following technical solutions:

[0007] A compound having a chemical structure as shown in Formula I or a pharmaceutically acceptable salt or solvate thereof:

[0008] wherein:

[0009] X is -CH2-, -O-, -NH- or -S-;

[0010] n = 0, 1 or 2;

[0011] R1 is a mono- or polysubstituted heterocyclic group, each substituent of the heterocyclic group being independently selected from -H, -C1-C6 alkyl, -C1-C6 haloalkyl, -C1-C6 alkoxy, -C1-C6 alkyl-O-C1-C6 alkyl, -C1-C6 alkyl-OH, -C1-C6 alkyl-NH2, -OH, -NH2, -COOH, -F, -Cl, -Br and -I;

[0012] A is wherein:

[0013] R2, R3 are -L(CH2) k C=OR4 or Z is -CH= or -N=, wherein:

[0014] L is -CH2-, -O-, -NH- or -S-; k = 1, 2 or 3; R4 is -NR5R6, R5, R6 are independently selected from -H, -C1-C5 alkyl, -C2-C6 alkenyl, -C2-C6 alkynyl, N and the attached R5, R6 do not form a ring or form a 4-7 membered heterocyclic ring; h = 1, 2, 3 or 4.

[0015] Preferably, the chemical structure of the above compound is shown as formula II:

[0016] wherein:

[0017] R1 is a mono- or poly-substituted heterocyclic group, each substituent of said heterocyclic group is independently selected from -H, -C1-C6 alkyl, -C1-C6 haloalkyl, -C1-C6 alkoxy, -C1-C6 alkyl-O-C1-C6 alkyl, -C1-C6 alkyl-OH, -C1-C6 alkyl-NH2, -OH, -NH2, -COOH, -F, -Cl, -Br and -I;

[0018] A is wherein:

[0019] R2, R3 are -L(CH2) k C=OR4 or Z is -CH= or -N=, wherein:

[0020] L is -CH2-, -O-, -NH- or -S-; k = 1, 2 or 3; R4 is -NR5R6, R5, R6 are independently selected from -H, -C1-C5 alkyl, -C2-C6 alkenyl, -C2-C6 alkynyl, N and the attached R5, R6 do not form a ring or form a 4-7 membered heterocyclic ring;

[0021] h = 1, 2, 3 or 4.

[0022] Preferably, R1 in the above chemical structure is shown as formula III:

[0023] wherein:

[0024] Y is -NH-, -O-, -S- or -C- substituted by R7 and R8, R7 and R8 are independently selected from -H, -OH and halogen;

[0025] R 1a , R 1b , R 1c , R 1d , R 1e , R 1f , R 1g , R 1hare each independently selected from the group consisting of -H, -Ci-C6alkyl, -Ci-C6haloalkyl, -Ci-C6alkoxy, -Ci-C6alkyl-O-Ci-C6alkyl, -Ci-C6alkyl-OH, -Ci-C6alkyl-NH2, -OH, -NH2, -COOH, -F, -CI, -Br, and -I.

[0026] More preferably, the chemical structure of the compound is selected from the group consisting of:

[0027] Use of any of the above compounds or pharmaceutically acceptable salts, solvates thereof for the manufacture of a BCL6 protein degrader.

[0028] Use of any of the above compounds or pharmaceutically acceptable salts, solvates thereof for the manufacture of a medicament for treating a disease that is treated or ameliorated by degrading BCL6 protein.

[0029] Preferably, the disease is cancer.

[0030] More preferably, the cancer is Hodgkin lymphoma, B-cell derived non-Hodgkin lymphoma, T-cell derived non-Hodgkin lymphoma, NK / T-cell derived non-Hodgkin lymphoma, or diffuse large B-cell lymphoma.

[0031] A method of treating a disease, administering to an individual having the disease a therapeutically effective amount of any of the above compounds or pharmaceutically acceptable salts, solvates thereof; wherein the disease is a disease that is treated or ameliorated by degrading BCL6 protein.

[0032] Preferably, the disease is cancer.

[0033] More preferably, the cancer is Hodgkin lymphoma, B-cell derived non-Hodgkin lymphoma, T-cell derived non-Hodgkin lymphoma, NK / T-cell derived non-Hodgkin lymphoma, or diffuse large B-cell lymphoma.

[0034] A pharmaceutical composition comprising any of the above compounds or pharmaceutically acceptable salts, solvates thereof.

[0035] Use of the above pharmaceutical composition for the manufacture of a medicament for treating a disease that is treated or ameliorated by degrading BCL6 protein.

[0036] Preferably, the disease is cancer.

[0037] More preferably, the cancer is Hodgkin lymphoma, B-cell derived non-Hodgkin lymphoma, T-cell derived non-Hodgkin lymphoma, NK / T-cell derived non-Hodgkin lymphoma, or diffuse large B-cell lymphoma.

[0038] A method of treating a disease, administering to an individual having the disease a therapeutically effective amount of a pharmaceutical composition comprising any of the above compounds or a pharmaceutically acceptable salt, solvate thereof; wherein the disease is a disease treated or ameliorated by degrading BCL6 protein.

[0039] Preferably, the disease is cancer.

[0040] More preferably, the cancer is Hodgkin lymphoma, B-cell derived non-Hodgkin lymphoma, T-cell derived non-Hodgkin lymphoma, NK / T-cell derived non-Hodgkin lymphoma or diffuse large B-cell lymphoma. Beneficial effects:

[0041] The tricyclic compound or a pharmaceutically acceptable salt, solvate thereof provided by the present application is novel in structure and has a relatively excellent degrading effect on BCL6 protein. Therefore, the compound provided by the present application or the composition containing the compound provided by the present application has the prospect of being developed into a drug for treating those diseases treated or ameliorated by degrading BCL6 protein, such as cancer, e.g. Hodgkin lymphoma, B-cell derived non-Hodgkin lymphoma, T-cell derived non-Hodgkin lymphoma, NK / T-cell derived non-Hodgkin lymphoma or diffuse large B-cell lymphoma. DETAILED DESCRIPTION

[0042] The following will specifically introduce the substantial content of the present application in combination with the compounds, but the protection scope of the present application is not limited thereto.

[0043] Synthetic route 1:

[0044] Compounds 1-9 are synthesized according to synthetic route 1.

[0045] Synthetic route 1. Reagents and conditions: (a) 1,2-dibromoethane, K2CO3, DMF, r.t., 3h; (b) BBr3, DCM, 0°C, 4h; (c) Cs2CO3, CH3CN, 60°C, 1h; (d) KNO3, H2SO4, 0°C, 4h; (e) (diazomethyl)trimethylsilane, Et3N, EtOH, r.t., 12h; (f) 2-bromo-N-methylacetamide, Cs2CO3, DMF, r.t., 3h; (g) Fe, NH4Cl, EtOH, H2O, 80°C, 3h; (h) DIPEA, DMSO, 100°C, 3h; (i) DIPEA, DMSO, 100°C, 10h.

[0046] 1-(2-bromoethyl)-7-methoxyindoline-2,3-dione (SM-1)

[0047] 1-(2-bromoethyl)-7-methoxyindoline-2,3-dione(SM-1)

[0048] General synthesis method 1: A-1a (5.00 g, 28.22 mmol), 1,2-dibromoethane A-1b (10.6 g, 56.45 mmol), potassium carbonate (7.80 g, 56.45 mmol) were added to a round bottom flask, 35 mL of DMF solution was added, stirred at room temperature for 3 h, TLC monitoring reaction was completed. 350 mL of water was added and stirred for half an hour, suction filtration, water washing, drying to get red solid SM-1 (6.80 g, yield: 84.8%). 1 H NMR (300 MHz, DMSO-d6) δ 7.45 (d, J = 8.1 Hz, 1H), 7.16 (dt, J = 15.4, 7.4 Hz, 2H), 4.20 (t, J = 6.9 Hz, 2H), 3.91 (s, 3H), 3.66 (t, J = 6.8 Hz, 2H).

[0049] 1-(2-bromoethyl)-7-hydroxyindoline-2,3-dione(SM-2)

[0050] 1-(2-bromoethyl)-7-hydroxyindoline-2,3-dione(SM-2)

[0051] General synthesis method 2: SM-1 (0.30 g, 1.06 mmol) was added to a round bottom flask containing 6 mL of dichloromethane solution, ice bath for 30 min, then slowly drop boron tribromide (0.79 g, 3.17 mmol), after 1 h, remove the ice bath, react at room temperature for 2 h, TLC monitoring reaction was completed. The reaction was ice bathed for half an hour, and anhydrous methanol was slowly added. When there was no white smoke produced, the drop was stopped, and the residue was concentrated under low pressure and purified by column chromatography (elution system: petroleum ether: ethyl acetate = 4:1, v / v) to obtain intermediate SM-2 as a red solid (0.16 g, yield: 56.1%). 1 H NMR (300 MHz, DMSO-d6) δ 10.48 (s, 1H), 7.17 (dd, J = 8.0, 1.3 Hz, 1H), 7.09 (dd, J = 7.4, 1.3 Hz, 1H), 7.03 - 6.95 (m, 1H), 4.21 (t, J = 6.9 Hz, 2H), 3.70 (t, J = 6.9 Hz, 2H).

[0052] 2,3-dihydro-[1,4]oxazino[2,3,4-hi]indole-5,6-dione(SM-3)

[0053] 2,3-dihydro-[1,4]oxazino[2,3,4-hi]indole-5,6-dione(SM-3)

[0054] Synthetic Route 3: SM-2 (1.00 g, 3.70 mmol) was added to a round bottom flask, 30 mL of acetonitrile solution was added, followed by cesium carbonate (2.41 g, 7.41 mmol), heated at 60 °C for 1 h, TLC monitored the end of the reaction. Remove the heating, let the reaction solution cool to room temperature, suction filtration, washed with ethyl acetate, concentrated, the residue was purified by column chromatography (eluent system: petroleum ether: ethyl acetate = 4: 1, v / v) to give intermediate SM-3 as a red solid (0.57 g, yield: 81.8%). 1 H NMR (300 MHz, DMSO-d6) δ 7.24 - 7.11 (m, 2H), 6.99 (t, J = 7.8 Hz, 1H), 4.32 (t, J = 4.7 Hz, 2H), 3.79 (t, J = 4.9 Hz, 2H).

[0055] 2,3-dihydro-[1,4]oxazino[2,3,4-hi]indole-5,6-dione(SM-4)

[0056] 2,3-dihydro-[1,4]oxazino[2,3,4-hi]indole-5,6-dione(SM-4)

[0057] According to synthetic route 1, SM-3 (2.40 g, 12.69 mmol) was added to a round bottom flask, stirred in ice bath for 15 min, then slowly added 20 mL of concentrated sulfuric acid solution, stirred in ice bath for 0.5 h, then slowly added potassium nitrate (1.28 g, 12.69 mmol), reacted at room temperature for 6 h, TLC monitored the end of the reaction. The reaction solution was slowly poured into stirring ice water, ethyl acetate was added, extracted three times, the organic phase was combined, concentrated under reduced pressure, the residue was purified by column chromatography (eluent system: petroleum ether: ethyl acetate = 4: 1, v / v) to give intermediate SM-4 as an orange yellow solid (1.80 g, yield: 60.6%). 1 H NMR (300 MHz, DMSO-d6) δ 8.06 (d, J = 2.0 Hz, 1H), 7.98 (d, J = 2.0 Hz, 1H), 4.41 (t, J = 4.7 Hz, 2H), 3.86 (t, J = 4.6 Hz, 2H).

[0058] 6-hydroxy-9-nitro-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-5-one (SM-5)

[0059] 6-hydroxy-9-nitro-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-5-one(SM-5)

[0060] According to synthetic route 1, SM-4 (0.33 g, 1.41 mmol) was added to a round bottom flask, triethylamine (0.28 g, 2.82 mmol) was added, followed by anhydrous ethanol (10 mL), then TMS-CHN2(0.70 mL, 1.40 mmol, 2M in hexane) was added dropwise slowly under stirring, argon was replaced, stirred at room temperature overnight, TLC monitored the end of the reaction. Concentrated under reduced pressure, the residue was purified by column chromatography (elution system dichloromethane:methanol = 80:1, v / v) to obtain intermediate SM-5 as a yellow solid (0.07 g, yield: 19.4%). 1 H NMR (300 MHz, DMSO-d6) δ 8.06 (d, J = 2.0 Hz, 1H), 7.98 (d, J = 2.0 Hz, 1H), 4.41 (t, J = 4.7 Hz, 2H), 3.86 (t, J = 4.6 Hz, 2H). ESI-MS: m / z: [M+H] + 249.05.

[0061] N-methyl-2-((9-nitro-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-6-yl)oxy)acetamide(SM-6a)

[0062] N-methyl-2-((9-nitro-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-6-yl)oxy)acetamide(SM-6a)

[0063] According to synthetic route 4, SM-5 (1.00 g, 4.03 mmol) was added to a round bottom flask, followed by SM-5a (0.74 g, 4.83 mmol), and cesium carbonate (2.63 g, 8.06 mmol), then DMF (15 mL) was added, stirred at room temperature for 1 h, TLC monitored the end of the reaction. An equal volume of ethyl acetate and water was added to extract 3 times, the organic layer was combined and concentrated under reduced pressure to remove the solvent, the residue was purified by column chromatography (elution system dichloromethane:methanol = 60:1, v / v) to obtain intermediate SM-6a as a yellow solid (0.84 g, yield: 65.5%). 1H NMR (300MHz, DMSO-d6) δ8.27(d,J=2.5Hz,1H),7.96(s,1H),7.75(d,J=2.5Hz,1H),7.54( s, 1H), 4.62 (s, 2H), 4.48 (t, J = 4.8Hz, 2H), 4.25 (t, J = 4.8Hz, 2H), 2.69 (d, J = 4.6Hz, 3H).

[0064] N,N-Dimethyl-2-((9-nitro-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-6-yl)oxy)acetamide (SM-6b)

[0065] N,N-dimethyl-2-((9-nitro-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-6-yl)oxy)acetamide(SM-6b)

[0066] Following general synthesis method 4, using SM-5 (1.00 g, 4.03 mmol) as the reactant, the target product, a yellow solid (0.84 g, yield: 65.5%), was obtained. 1 H NMR(300MHz,DMSO-d6)δ8.23(d,J=2.5Hz,1H),7.73(d,J=2.5Hz,1H),7.45(s,1H),4 .97(s,2H),4.49(t,J=4.7Hz,2H),4.24(t,J=4.8Hz,2H),3.05(s,3H),2.90(s,3H).

[0067] 2-(9-amino-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-6-yl)oxy)-N-methylacetamide (SM-7a)

[0068] 2-((9-amino-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-6-yl)oxy)-N-methylacetamide(SM-7a)

[0069] Synthetic General Procedure 5: SM-6a (0.40 g, 1.25 mmol) was taken in a round bottom flask containing absolute ethanol: water = 20 mL: 4 mL, ammonium chloride (0.35 g, 6.26 mmol) was added, under stirring iron powder (0.34 g, 6.26 mmol) was added, the reaction was heated at 80 °C for 3 h, TLC was monitored for completion of reaction. The heating was removed and the reaction was allowed to cool to room temperature, 2 mL of ammonium hydroxide solution was added to adjust the pH of the solution to more than 7, filtered through celite, washed with dichloromethane: methanol = 20: 1 (600 mL), the filtrate was collected and concentrated under reduced pressure, extracted with ethyl acetate and concentrated under reduced pressure to get SM-7a (0.24 g, yield: 67.0%) as a light yellow solid. ESI-MS: m / z: [M+H] + 290.11.

[0070] 2-((9-((2,5-dichloropyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-6-yl)oxy)-N-methylacetamide(SM-9a)

[0071] 2-((9-((2,5-dichloropyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-6-yl)oxy)-N-methylacetamide(SM-9a)

[0072] Synthetic General Procedure 5: SM-6a (0.40 g, 1.25 mmol) was taken in a round bottom flask containing absolute ethanol: water = 20 mL: 4 mL, ammonium chloride (0.35 g, 6.26 mmol) was added, under stirring iron powder (0.34 g, 6.26 mmol) was added, the reaction was heated at 80 °C for 3 h, TLC was monitored for completion of reaction. The heating was removed and the reaction was allowed to cool to room temperature, 2 mL of ammonium hydroxide solution was added to adjust the pH of the solution to more than 7, filtered through celite, washed with dichloromethane: methanol = 20: 1 (600 mL), the filtrate was collected and concentrated under reduced pressure, extracted with ethyl acetate and concentrated under reduced pressure to get SM-7a (0.24 g, yield: 67.0%) as a light yellow solid. ESI-MS: m / z: [M+H] + 304.12.

[0073] 2-((9-((2,5-dichloropyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-6-yl)oxy)-N-methylacetamide(SM-9a)

[0074] 2-((9-((2,5-dichloropyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-6-yl)oxy)-N-methylacetamide(SM-9a)

[0075] Synthetic General Procedure 6: SM-7a (0.40 g, 1.38 mmol), SM-8 (0.51 g, 2.77 mmol) were added to a round bottom flask containing 20 mL of DMSO, DIPEA (0.54 g, 4.15 mmol) was added, heated at 100 °C for 3 h, TLC monitored the completion of reaction. After the reaction was cooled to room temperature, 80 mL of water was added and stirred for 30 min, suction filtered, washed with water, oven dried to get yellow solid SM-9a (0.43 g, yield: 71.6 %). ESI-MS: m / z: [M+H] + 436.04.

[0076] 2-((9-((2,5-dichloropyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-6-yl)oxy)-N,N-dimethylacetamide(SM-9b)

[0077] 2-((9-((2,5-dichloropyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-6-yl)oxy)-N,N-dimethylacetamide(SM-9b)

[0078] Following synthetic general procedure 6, SM-7b (0.40 g, 1.38 mmol), SM-8 (0.51 g, 2.77 mmol) as starting materials to get the target product SM-9b (0.43 g, yield: 76.0 %) as yellow solid. ESI-MS: m / z: [M+H] + 450.06.

[0079] Example 1

[0080] 2-(9-(5-chloro-2-(3R,5S)-3,5-dimethylpiperidin-1-yl)pyrimidin-4-amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-6-yl)oxy)-N-methylacetamide (1)

[0081] 2-(9-(5-chloro-2-(3R,5S)-3,5-dimethylpiperidin-1-yl)pyrimidin-4-amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-6-yl)oxy)-N-methylacetamide (1)

[0082] General procedure 7: SM-9a (0.05 mg, 0.12 mmol), SM-10a (0.03 g, 0.23 mmol) were added to a round bottom flask containing 4 mL of DMSO, DIPEA (0.05 g, 0.34 mmol) was added, heated at 100 °C for 10 h, TLC monitored the reaction was completed. After the reaction was cooled to room temperature, an equal volume of ethyl acetate and water were added and extracted 3 times, the organic layer was combined and concentrated under low pressure to remove the solvent, the residue was column chromatography (elution system dichloromethane:methanol = 60:1, v / v) to obtain the product brown solid 1 (0.02 g, yield: 35.7%). 1 H NMR (300 MHz, DMSO-d6) δ 8.74 (s, 1H), 8.04 (s, 1H), 7.94 (s, 1H), 7.52 (d, J = 2.2 Hz, 1H), 7.47 (d, J = 2.2 Hz, 1H), 7.15 (s, 1H), 4.57 (s, 2H), 4.51 (d, J = 12.9 Hz, 2H), 4.40 (t, J = 4.7 Hz, 2H), 4.19 (t, J = 4.8 Hz, 2H), 2.68 (d, J = 4.7 Hz, 3H), 2.31 (t, J = 12.1 Hz, 2H), 1.79 (d, J = 12.7 Hz, 1H), 1.55 (ddt, 2H), 0.89 (d, J = 6.6 Hz, 6H), 0.81 (q, J = 12.0 Hz, 1H). HRMS (ESI): calcd for C 25 H 29 ClN6O4[M+H] + 513.00, found 513.2040 purity: 94.99% by HPLC (MeOH / H2O = 80:20, t R = 5.932 min).

[0083] Example 2

[0084] 2-(9-(5-chloro-2-(2S,6R)-2,6-dimethylmorpholin-4-yl)pyrimidin-4-ylamino)-5- oxo-2,3-dihydro-5H-[l,4]oxazepino[2,3,4-ij]quinolin-6-yloxy)-N- methylacetamide (2)

[0085] 2-((9-((5-chloro-2-((2S,6R)-2,6-dimethylmorpholino)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-6-yl)oxy)-N-methylacetamide(2)

[0086] Following general synthesis 7, SM-9a (0.05 mg, 0.12 mmol) as the starting material, the target product 2 was obtained as a brown solid (18 mg, yield: 30.5%). 1 H NMR (300 MHz, DMSO-d6) δ 8.89 (s, 1H), 8.09 (s, 1H), 7.99 (s, 1H), 7.57 (d, J = 2.3 Hz, 1H), 7.40 (d, J = 2.1 Hz, 1H), 7.19 (s, 1H), 4.58 (s, 2H), 4.40 (s, 2H), 4.34 (d, J = 12.4 Hz, 2H), 4.19 (t, J = 4.8 Hz, 2H), 3.60 - 3.52 (m, 2H), 2.68 (d, J = 4.6 Hz, 3H), 2.48 (s, 2H), 1.13 (d, J = 6.1 Hz, 6H). HRMS (ESI): calcd for C 24 H 27 ClN6O5[M+H] + 515.1731, found 515.1804.

[0087] Example 3

[0088] (S)-2-((9-((5-chloro-2-(3-methylpiperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-6-yl)oxy)-N-methylacetamide(3)

[0089] (S)-2-((9-((5-chloro-2-(3-methylpiperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-6-yl)oxy)-N-methylacetamide(3)

[0090] Following the general procedure 7, SM-9a (0.05 mg, 0.12 mmol) was used as the starting material to obtain the target product 3 as a brown solid (23 mg, yield: 40.2%). 1 H NMR (300 MHz, DMSO-d6) d 8.75 (s, 1H), 8.01 (d, J = 11.5 Hz, 2H), 7.56 (s, 1H), 7.43 (s, 1H), 7.12 (s, 1H), 4.59 (s, 2H), 4.40 (s, 4H), 4.19 (s, 2H), 3.48 (s, 1H), 2.92 - 2.78 (m, 1H), 2.76 - 2.62 (m, 3H), 1.65 (d, J = 9.0 Hz, 1H), 1.26 (t, J = 26.5 Hz, 4H), 0.89 (d, J = 6.4 Hz, 3H). HRMS (ESI): calcd for C 24 H 27 ClN6O4[M+H] + 499.1782, found 499.1848.

[0091] Example 4

[0092] 2-((9-((5-chloro-2-(4-fluoropiperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-6-yl)oxy)-N-methylacetamide(4)

[0093] 2-((9-((5-chloro-2-(4-fluoropiperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-6-yl)oxy)-N-methylacetamide(4)

[0094] Following the general procedure 7, SM-9a (0.05 mg, 0.12 mmol) was used as the starting material to obtain the target product 3 as a brown solid (23 mg, yield: 40.2%). 1H NMR (300 MHz, DMSO-d6) δ 8.97 (s, 1H), 8.22 (s, 1H), 8.16 (t, J = 6.0 Hz, 1H), 7.73 (d, J = 2.2 Hz, 1H), 7.49 (d, J = 2.2 Hz, 1H), 7.24 (s, 1H), 5.05 (d, J = 48.6 Hz, 1H), 4.75 (s, 2H), 4.54 (t, J = 4.7 Hz, 2H), 4.32 (t, J = 4.6 Hz, 2H), 3.96 (s, 2H), 3.80 (s, 2H), 2.81 (d, J = 4.6 Hz, 3H), 2.06 (s, 2H), 1.84 (s, 2H). HRMS (ESI): calcd for C 23 H 24 ClFN6O4[M+H] + 503.1532, found 503.1603.

[0095] Example 5

[0096] 2-((9-((5-chloro-2-(4-chloropiperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-6-yl)oxy)-N-methylacetamide(5)

[0097] 2-((9-((5-chloro-2-(4-chloropiperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-6-yl)oxy)-N-methylacetamide(5)

[0098] Following General Synthesis 7, SM-9a (0.05 mg, 0.12 mmol) was used as the starting material to give the target product 5 as a brown solid (19 mg, yield: 31.9%). 1H NMR (300 MHz, DMSO-d6) δ 8.82 (s, 1H), 8.06 (s, 1H), 8.00 (d, J = 4.8 Hz, 1H), 7.56 (d, J = 2.2 Hz, 1H), 7.34 (d, J = 2.2 Hz, 1H), 7.09 (s, 1H), 4.59 (s, 2H), 4.46 (s, 1H), 4.38 (t, J = 4.8 Hz, 2H), 4.17 (d, J = 5.5 Hz, 2H), 4.00 (d, J = 13.3 Hz, 2H), 3.50 (d, J = 10.3 Hz, 2H), 2.66 (d, J = 4.6 Hz, 3H), 2.07 (d, J = 11.2 Hz, 2H), 1.77 - 1.67 (m, 2H). HRMS (ESI): calcd for C 23 H 24 Cl2N6O4[M+H] + 519.1236, found 519.1289.

[0099] Example 6

[0100] 2-((9-((5-chloro-2-(4,4-difluoropiperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-6-yl)oxy)-N-methylacetamide(6)

[0101] 2-((9-((5-chloro-2-(4,4-difluoropiperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-6-yl)oxy)-N-methylacetamide(6)

[0102] Following General Synthesis 7, SM-9a (0.05 mg, 0.12 mmol) as the starting material, the target product 6 was obtained as a brown solid (14 mg, yield: 23.4%). 1H NMR (300 MHz, DMSO-d6) δ 8.89 (s, 1H), 8.11 (s, 1H), 8.00 (d, J = 5.0 Hz, 1H), 7.59 (d, J = 2.2 Hz, 1H), 7.31 (d, J = 2.1 Hz, 1H), 7.11 (s, 1H), 4.61 (s, 2H), 4.39 (t, J = 5.3 Hz, 2H), 4.19 (t, J = 4.7 Hz, 2H), 3.79 (d, J = 5.9 Hz, 4H), 2.67 (d, J = 4.6 Hz, 3H), 2.02 (d, J = 15.3 Hz, 4H). HRMS (ESI): calcd for C 23 H 23 ClF2N6O4[M+H] + 521.1437, found 521.1509.

[0103] Example 7

[0104] (R)-2-((9-((5-chloro-2-(3-methylpiperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-6-yl)oxy)-N-methylacetamide(7)

[0105] (R)-2-((9-((5-chloro-2-(3-methylpiperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-6-yl)oxy)-N-methylacetamide(7)

[0106] Following General Synthesis 7, SM-9a (0.05 mg, 0.12 mmol) as the starting material, the target product 7 was obtained as a brown solid (15 mg, yield: 26.2%). 1H NMR (300 MHz, DMSO-d6) δ 8.75 (s, 1H), 8.04 (s, 1H), 7.98 (d, J = 5.3 Hz, 1H), 7.56 (d, J = 2.1 Hz, 1H), 7.44 (d, J = 2.0 Hz, 1H), 7.12 (s, 1H), 4.59 (s, 2H), 4.39 (d, J = 5.9 Hz, 4H), 4.19 (d, J = 5.0 Hz, 2H), 2.84 (t, J = 12.2 Hz, 1H), 2.68 (d, J = 4.6 Hz, 3H), 2.57 (s, 1H), 1.26 (d, J = 7.5 Hz, 4H), 0.90 (d, J = 6.5 Hz, 3H). HRMS (ESI): calcd for C 24 H 27 ClN6O4[M+H] + 499.1782, found 499.1845.

[0107] Example 8

[0108] 2-((9-((5-chloro-2-(3-(hydroxymethyl)piperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-6-yl)oxy)-N-methylacetamide(8)

[0109] 2-((9-((5-chloro-2-(3-(hydroxymethyl)piperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-6-yl)oxy)-N-methylacetamide(8)

[0110] Following General Synthesis 7, SM-9a (0.05 mg, 0.12 mmol) as the starting material, the target product 8 was obtained as a brown solid (22 mg, yield: 37.3%). 1H NMR(300MHz,Chloroform-d)δ8.13(s,1H),7.80(s,1H),7.15(m,2H),6.69(m,2H),6 .25(d,J=12.3Hz,1H),5.01(m,2H),4.76–4.67(d,2H),4.51(m,2H),3.64(m,J=12.5 ,1H),3.59(m,2H),3.32(m,J=14.1Hz,1H),3.07(m,J=7.0Hz,1H),2.83(d,3H),1.82 (m,1H),1.55(m,3H),1.48(m,J=13.2,1H),1.05(m,J=11.7Hz,1H).HRMS(ESI):calcd for C 24 H 27 ClN6O5[M+H] + 515.1731, found 515.1795.

[0111] Example 9

[0112] 2-((9-((5-chloro-2-((3R,5S)-3,5-dimethylpiperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-6-yl)oxy)-N,N-dimethylacetamide(9)

[0113] 2-((9-((5-chloro-2-((3R,5S)-3,5-dimethylpiperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-6-yl)oxy)-N,N-dimethylacetamide(9)

[0114] Following general synthesis method 7, using SM-9b (0.05 g, 0.11 mmol) as the reactant, the target product, brown solid 9 (25 mg, yield: 42.7%), was obtained. 1H NMR (300 MHz, Chloroform-d) δ 7.99 (s, 1H), 7.49 (d, J = 2.1 Hz, 1H), 7.30 (d, J = 1.8 Hz, 1H), 7.10 (s, 1H), 4.85 (s, 2H), 4.62 (d, J = 12.8 Hz, 2H), 4.39 (d, J = 5.6 Hz, 2H), 4.32 (d, J = 5.0 Hz, 2H), 3.14 (s, 3H), 2.96 (s, 3H), 2.35 (t, J = 12.1 Hz, 2H), 1.85 (d, J = 13.0 Hz, 1H), 1.69 - 1.64 (m, 2H), 1.35 (d, J = 11.7 Hz, 1H), 0.96 (d, J = 6.5 Hz, 6H). HRMS (ESI): calcd for C 26 H 31 ClN6O4[M+H] + 527.2095, found 527.2170.

[0115] Synthesis Route 2:

[0116] Compound 10 was synthesized according to Synthesis Route 2.

[0117] Synthesis Route 2. Reagents and conditions: (a) 1,2-dibromoethane, K2CO3, DMF, r.t., 3h; (b) Pd / C, H2, MeOH, r.t., 4h; (c) CDI, DMF, r.t., 2h; (d) BBr3, DCM, 0°C, 4h; (e) Cs2CO3, CH3CN, 60°C, 1h; (f) BnNH2, LiHMDS, BrettPhos Pd G3, N2, THF, 80°C, 2h; (g) Pd / C, H2, MeOH, r.t., 4h; (h) DIPEA, DMSO, 20°C, 3h; (i) DIPEA, DMSO, 100°C, 10h; (f) KI, Cs2CO3, DMSO, 80°C, 12h.

[0118] 4-bromo-N-(2-bromoethyl)-2-methoxy-6-nitroaniline (SM-11)

[0119] 4-bromo-N-(2-bromoethyl)-2-methoxy-6-nitroaniline (SM-11)

[0120] Following General Synthesis Procedure 1, using B-1 (5.00 g, 20.24 mmol), 1,2-dibromoethane A-1b (4.90 g, 26.31 mmol) as starting materials, the target product, yellow solid SM-11 (4.2 g, yield: 59.0%) was obtained. 1 H NMR (300 MHz, Chloroform-d) δ 6.85 (d, J = 2.0 Hz, 1H), 6.76 (d, J = 2.0 Hz, 1H), 3.94 (s, 3H), 3.60 - 3.49 (m, 4H).

[0121] 4-bromo-N 1 -(2- bromoethyl)-6-methoxybenzene-1,2-diamine (SM-12)

[0122] 4-bromo-N 1 -(2- bromoethyl)-6-methoxybenzene-1,2-diamine (SM-12)

[0123] Following Synthetic Route 2, SM-11 (4.20 g, 11.86 mmol) was added to a round bottom flask containing 30 mL of methanol solution, Pd / C (10%, 0.30 g) was added, hydrogen was replaced, stirred at room temperature for 4 h, TLC monitored the end of the reaction. Silica gel was filtered, washed with methanol, the organic phase was combined and concentrated under reduced pressure to obtain the intermediate SM-12 as a light yellow solid (3.62 g, yield: 94.2%). ESI-MS: m / z: [M+H] + 322.95.

[0124] 5-bromo-1-(2-bromoethyl)-7-methoxy-1,3-dihydro-2H-benzo[d]imidazol-2-one (SM-13)

[0125] 5-bromo-1-(2-bromoethyl)-7-methoxy-1,3-dihydro-2H-benzo[d]imidazol-2-one (SM-13)

[0126] Following Synthetic Route 2, SM-12 (3.00 g, 9.26 mmol) was added to a round bottom flask containing 30 mL of DMF solution, CDI (2.61 g, 18.52 mmol) was added, stirred at room temperature for 2 h, TLC monitored the end of the reaction. An equal volume of ethyl acetate and water was added and extracted 3 times, the organic layer was combined and concentrated under reduced pressure to remove the solvent, the residue was purified by column chromatography (elution system: dichloromethane:methanol = 60:1, v / v) to obtain the intermediate SM-13 as a yellow solid (1.81 g, yield: 55.8%).1 HNMR (300 MHz, Chloroform-d) δ 7.05 (d, J = 1.5 Hz, 1H), 6.79 (d, J = 1.5 Hz, 1H), 3.94 (s, 3H), 3.60 - 3.53 (m, 2H), 3.46 - 3.39 (m, 2H).

[0127] 5-bromo-1-(2-bromoethyl)-7-hydroxy-1,3-dihydro-2H-benzo[d]imidazol-2-one (SM-14)

[0128] 5-bromo-1-(2-bromoethyl)-7-hydroxy-1,3-dihydro-2H-benzo[d]imidazol-2-one (SM-14)

[0129] Following the general synthesis procedure 2, SM-13 (1.50 g, 4.29 mmol) was used as a starting material to obtain the target product SM-14 (1.02 g, yield: 70.8%) as a yellow solid. 1 HNMR (300 MHz, Chloroform-d) δ 7.05 (d, J = 1.5 Hz, 1H), 6.79 (d, J = 1.5 Hz, 1H), 3.94 (s, 3H), 3.60 - 3.53 (m, 2H), 3.46 - 3.39 (m, 2H).

[0130] 7-bromo-3,4-dihydro-5-oxa-1,2a-diazaacenaphthylen-2(1H)-one (SM-15)

[0131] 7-bromo-3,4-dihydro-5-oxa-1,2a-diazaacenaphthylen-2(1H)-one (SM-15)

[0132] Following the general synthesis procedure 3, SM-14 (1.50 g, 4.29 mmol) was used as a starting material to obtain the target product SM-15 (0.51 g, yield: 67.6%) as a yellow solid. 1 HNMR (300 MHz, Chloroform-d) δ 7.05 (d, J = 1.5 Hz, 1H), 6.79 (d, J = 1.5 Hz, 1H), 3.94 (s, 3H), 3.60 - 3.53 (m, 2H), 3.46 - 3.39 (m, 2H).

[0133] 7-bromo-3,4-dihydro-5-oxa-1,2a-diazaacenaphthylen-2(1H)-one (SM-15)

[0134] 7-(benzylamino)-3,4-dihydro-5-oxa-1,2a-diazaacenaphthylen-2(1H)-one (SM-16)

[0135] According to synthetic route 2, SM-15 (0.50 g, 1.96 mmol) was added to a sealed tube containing 10 mL of DMF solution, LiHMDS (1 M in THF, 4.70 mL, 4.70 mmol) was added dropwise slowly at room temperature, argon replacement, then BrettPhos (0.04 g, 0.08 mmol) BrettPhos Pd G3 (0.04 g, 0.04 mmol) were added in turn, argon replacement, 80 °C stirring for 2 h, TLC monitoring reaction end. The reaction mixture was extracted with equal volume of ethyl acetate and water for 3 times, the organic phase was combined and concentrated under reduced pressure to remove the solvent, and the residue was purified by column chromatography (eluent system: dichloromethane:methanol = 100:1, v / v) to obtain intermediate SM-16 as a yellow solid (0.36 g, yield: 65.5%). 1 H NMR (300 MHz, Chloroform-d) δ 7.41 - 7.29 (m, 5H), 6.74 (d, J = 1.5 Hz, 1H), 5.58 (d, J = 1.7 Hz, 1H), 4.32 (d, 2H), 4.20 (t, J = 4.9 Hz, 2H), 3.35 (t, J = 4.9 Hz, 2H).

[0136] 7-amino-3,4-dihydro-5-oxa-1,2a-diazaacenaphthylen-2(1H)-one (SM-17)

[0137] 7-amino-3,4-dihydro-5-oxa-1,2a-diazaacenaphthylen-2(1H)-one (SM-17)

[0138] According to synthetic route 2, SM-16 (0.80 g, 2.84 mmol) was added to a round bottom flask containing 10 mL of methanol solution, Pd / C (10%, 0.12 g) was added, hydrogen replacement, stirring at room temperature for 4 h, TLC monitoring reaction end. Silica gel filtration, methanol washing, the organic phase was combined and concentrated under reduced pressure, and the residue was purified by column chromatography (eluent system: dichloromethane:methanol = 70:1, v / v) to obtain intermediate SM-17 as a yellow solid (0.49 g, yield: 90.1%). ESI-MS: m / z: [M+H] + 192.10.

[0139] 7-((2,5-dichloropyrimidin-4-yl)amino)-3,4-dihydro-5-oxa-1,2a-diazaacenaphthylen-2(1H)-one (SM-18)

[0140] 7-((2,5-dichloropyrimidin-4-yl)amino)-3,4-dihydro-5-oxa-1,2a-diazaacenaphthylen-2(1H)-one (SM-18)

[0141] Following General Synthesis 6, SM-17 (1.00 g, 5.23 mmol) was used as the starting material to obtain the target product SM-18 (1.26 g, yield: 71.2%) as a brown solid. ESI-MS: m / z: [M+H] + 338.05.

[0142] 7-((5-chloro-2-((3R,5S)-3,5-dimethylpiperidin-1-yl)pyrimidin-4-yl)amino)-3,4-dihydro-5-oxa-1,2a-diazaacenaphthylen-2(1H)-one (SM-19a)

[0143] 7-((5-chloro-2-((3R,5S)-3,5-dimethylpiperidin-1-yl)pyrimidin-4-yl)amino)-3,4-dihydro-5-oxa-1,2a-diazaacenaphthylen-2(1H)-one (SM-19a)

[0144] Following General Synthesis 7, SM-18 (0.20 g, 0.59 mmol) was used as the starting material to obtain the target product SM-19a (0.13 g, yield: 53.0%) as a brown solid. 1 H NMR (300 MHz, Chloroform-d) δ 8.17 (s, 1H), 6.81 (d, J = 1.5 Hz, 1H), 6.54 (d, J = 1.5 Hz, 1H), 4.39 (m, J = 6.0 Hz, 4H), 4.19 (t, J = 5.0 Hz, 2H), 3.67 (m, J = 12.4, 7.0 Hz, 2H), 1.81 - 1.68 (m, J = 6.9 Hz, 2H), 1.66 (m, 1H), 0.98 (m, 7H).

[0145] Example 10

[0146] 7-((5-chloro-2-((3R,5S)-3,5-dimethylpiperidin-1-yl)pyrimidin-4-yl)amino)-1-(3-hydroxy-3-methylbutyl)-3,4-dihydro-5-oxa-1,2a-diazaacenaphthylen-2(1H)-one (10)

[0147] 7-((5-chloro-2-((3R,5S)-3,5-dimethylpiperidin-1-yl)pyrimidin-4-yl)amino)-1-(3-hydroxy-3-methylbutyl)-3,4-dihydro-5-oxa-1,2a-diazaacenaphthylen-2(1H)-one (10)

[0148] According to synthetic route 2, SM-19a (0.10 g, 0.24 mmol), SM-19b (0.12 g, 0.48 mmol) were added to a round bottom flask containing 5 mL of DMSO, potassium iodide (0.02 g, 0.12 mmol) and cesium carbonate (0.16 g, 0.48 mmol) were added, heated at 80 °C for 12 h, TLC monitored the end of the reaction. After the reaction was cooled to room temperature, an equal volume of ethyl acetate and water were added and extracted 3 times, the organic layer was combined and concentrated under low pressure to remove the solvent, the residue was column chromatography (elution system dichloromethane:methanol = 40:1, v / v) to obtain the product brown solid 10 (0.03 g, yield: 24.8%). 1 H NMR (300 MHz, Chloroform-d) δ 8.19 (s, 1H), 6.81 (d, J = 2.0 Hz, 1H), 6.54 (d, J = 2.0 Hz, 1H), 4.28 (m, 4H), 4.15 (t, J = 5.0 Hz, 2H), 3.68 (m, 4H), 1.86 (m, J = 13.4, 2H), 1.74 (m, J = 13.7, 2H), 1.66 - 1.57 (m, 1H), 1.29 (s, 6H), 0.98 - 0.86 (m, 7H). HRMS (ESI): calcd for C 25 H 29 ClN6O4[M+H] + 501.23, found 501.2324 purity: 96.91% by HPLC (MeOH / H2O = 80:20, t R = 4.671 min).

[0149] Pharmacological activity evaluation:

[0150] OCI-LY1 cells (Zhejiang Meisen Cell Technology Co., Ltd.) were inoculated in a 6-well plate, and then different concentrations of compounds were added. After 12 hours of administration, the cells were centrifuged, mixed and lysed with medium-strength RIPA lysis buffer (Bi Yun Tian Biotechnology), the supernatant was aspirated after centrifugation, and the protein concentration was determined by BCA method. The protein sample was mixed with protein loading buffer (Bi Yun Tian Biotechnology) and heated at 100°C for 10 min to prepare the sample, then the sample was added to 12% polyacrylamide gel SDS-PAGE, electrophoresis at 60V constant voltage until the marker left the concentrated gel, then continue electrophoresis at 120V constant voltage, transfer the membrane in wet transfer buffer containing 10% methanol for 90 min, then cut the required band from the transferred PVDF membrane, block with milk for 2 hours, add anti-BCL6 antibody (abcam) and β-Actin (Proteintech) antibody diluted with milk in the corresponding band, incubate at 4°C overnight. The next day, wash the primary antibody with TBST, add the secondary antibody and incubate at room temperature for 45 min. Continue to wash the secondary antibody solution with TBST, and scan the membrane under the Odyssey Infrared Imaging System (LI-COR, Lincoln, Nebraska, USA). DC 50 The concentration required for the BCL6 degradation agent to degrade 50% of the BCL6 protein is referred to as the degradation concentration (DC). The calculation process is as follows: the degradation agent is diluted at 9 concentrations with 5-fold gradient from 1000 nM, and the degradation of BCL6 protein at each concentration is detected by WB method. Image J software is used for gray scale analysis to calculate the remaining amount of BCL6. The protein remaining amount and the concentration logarithm (Log(C)) are fitted by Graphpad 8.0 software to obtain the DC 50 value.

[0151] Table 1 Degradation of BCL6 in OCI-LY1 cells by compounds

Claims

1. A compound with a chemical structure as shown in Formula I, or a pharmaceutically acceptable salt or solvate thereof: wherein: X is -CH2-, -O-, -NH- or -S-; n = 0, 1 or 2; R1 is a mono- or poly-substituted heterocyclyl, each substituent of said heterocyclyl being independently selected from the group consisting of -H, -Ci-C6alkyl, -Ci-C6haloalkyl, -Ci-C6alkoxy, -Ci-C6alkyl-O-Ci-C6alkyl, -Ci-C6alkyl-OH, -Ci-C6alkyl-NH2, -OH, -NH2, -COOH, -F, -Cl, -Br and -I; A is wherein: R2, R3 are -L(CH2) k C=OR4or Z is -CH= or -N=, wherein: L is -CH2-, -O-, -NH- or -S-; k = 1, 2 or 3; R4 is -NR5R6, R5, R6 are independently selected from the group consisting of -H, -Ci-C5alkyl, -C2-C6alkenyl, -C2-C6alkynyl, N and the attached R5, R6 do not form a ring or form a 4-7 membered heterocyclic ring; h = 1, 2, 3 or 4.

2. The compound of claim 1, or a pharmaceutically acceptable salt, solvate thereof, wherein, The chemical structure is shown as Formula II: wherein: R1 is a mono- or poly-substituted heterocyclyl, each substituent of said heterocyclyl being independently selected from the group consisting of -H, -Ci-C6alkyl, -Ci-C6haloalkyl, -Ci-C6alkoxy, -Ci-C6alkyl-O-Ci-C6alkyl, -Ci-C6alkyl-OH, -Ci-C6alkyl-NH2, -OH, -NH2, -COOH, -F, -Cl, -Br and -I; A is wherein: R2, R3 are -L(CH2) k C=OR4or Z is -CH= or -N=, wherein: L is -CH2-, -O-, -NH- or -S-; k = 1, 2 or 3; R4 is -NR5R6, R5, R6 are independently selected from the group consisting of -H, -Ci-C5alkyl, -C2-C6alkenyl, -C2-C6alkynyl, N and the attached R5, R6 do not form a ring or form a 4-7 membered heterocyclic ring; h = 1, 2, 3 or 4.

3. The compound or pharmaceutically acceptable salt, solvate thereof according to claim 1 or 2, characterized in that, R1is as shown in Formula III: wherein: Y is -NH-, -O-, -S- or -C- substituted with R7 and R8, R7 and R8 are independently selected from the group consisting of -H, -OH and halogen; R 1a , R 1b , R 1c , R 1d , R 1e , R 1f , R 1g , R 1h are each independently selected from the group consisting of -H, -Ci-C6alkyl, -Ci-C6haloalkyl, -Ci-C6alkoxy, -Ci-C6alkyl-O-Ci-C6alkyl, -Ci-C6alkyl-OH, -Ci-C6alkyl-NH2, -OH, -NH2, -COOH, -F, -Cl, -Br, and -I.

4. The compound of claim 3, or a pharmaceutically acceptable salt, solvate thereof, wherein, the chemical structure of the compound is selected from:

5. Use of a compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt, solvate thereof, for the manufacture of a BCL6 protein degrader.

6. Use of a compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt, solvate thereof, for the manufacture of a medicament for the treatment of a disease which is treated or alleviated by the degradation of BCL6 protein.

7. Use according to claim 6, characterized in that: The disease is cancer.

8. Use according to claim 7, characterized in that: The cancer is Hodgkin lymphoma, B-cell derived non-Hodgkin lymphoma, T-cell derived non-Hodgkin lymphoma, NK / T-cell derived non-Hodgkin lymphoma or diffuse large B-cell lymphoma.

9. A pharmaceutical composition, characterized by: containing a compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt, solvate thereof.

10. Use of a pharmaceutical composition according to claim 9 for the manufacture of a medicament for the treatment of a disease which is treated or alleviated by the degradation of BCL6 protein.

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