WO2025242012A1 - Protein-targeting degradation chimeras and use thereof - Google Patents

Abstract

Disclosed are protein-targeting degradation chimeras and the use thereof. The protein-targeting degradation chimeras have a new chemical structure, exhibit a great degradation effect on the BCL6 protein, can be used for preparing BCL6 degrader drugs, and has the potential to be developed into a drug for treating diseases that can be treated or alleviated by means of the degradation of the BCL6 protein. The diseases that can be treated or alleviated by means of the degradation of the BCL6 protein comprise cancers such as Hodgkin lymphoma, B-cell-derived non-Hodgkin lymphoma, T-cell-derived non-Hodgkin lymphoma, NK/T-cell-derived non-Hodgkin lymphoma and diffuse large B-cell lymphoma.

Description

A class of protein-targeted degradation chimeras and their applications Technical Field

This invention belongs to the field of medicinal chemistry, specifically relating to a class of protein-targeted degradation chimeras and their uses.

Background Technology

B-cell lymphokine 6 (BCL6) plays a crucial role in humoral immune responses. It is a human gene of approximately 24 kb, encoding a protein of about 95 kDa. BCL6 is a member of the POZ/BTB/Zinc finger protein family and is primarily composed of three parts: 1) The N-terminal POZ/BTB domain: This is the main functional region responsible for transcriptional repression. When BCL6 exerts its transcriptional repression function, the BTB domain spontaneously forms a dimer. Three important transcriptional co-repressors, SMRT, BCOR, or NCOR, competitively bind to the BTB binding site, collectively exerting transcriptional repression. This process participates in the early transcriptional regulation of germinal centers (GCs). 2) The central region (also known as the RD2 domain) contains three PEST domains, whose main function is to recruit cofactors such as MTA3 or CTBP1, maintaining protein stability. 3) The C-terminal zinc finger domain consists of six identical zinc finger structures, primarily binding to DNA and is a prerequisite for BCL6's transcriptional repression function.

BCL6 is a transcriptional repressor that recruits co-repressors through its BTB domain. BCL6 protein is overexpressed in lymphoma, breast cancer, ovarian cancer, non-small cell lung cancer, and glioma, and is closely associated with poor patient prognosis. When BCL6 protein is highly expressed, the p53 tumor suppressor gene is almost absent. In GC responses, chromosomal translocations and point mutations of BCL6 lead to persistently high BCL6 protein expression, promoting malignant proliferation of B cells and resulting in B-cell lymphoma. Most non-Hodgkin lymphomas (NHL) originate from GC, with diffuse large B-cell lymphoma (DLBCL) being the most common subtype. BCL6 is considered a carcinogenic driver of DLBCL. Numerous preclinical studies have also shown that blocking the interaction between the BCL6-BTB domain and its transcriptional co-repressors can inhibit GC formation and NHL cell proliferation, representing an effective and safe treatment strategy for NHL without toxic side effects or macrophage-driven inflammatory responses.

BCL6 is a highly promising target for cancer therapy, including but not limited to Hodgkin's lymphoma, B-cell non-Hodgkin's lymphoma, T-cell non-Hodgkin's lymphoma, NK/T-cell non-Hodgkin's lymphoma, and diffuse large B-cell lymphoma. Currently, there are approximately 18 BCL6 drugs in development, categorized into small molecule inhibitors (8 classes), protein-targeted degradation chimeras (5 classes), and molecular gels (5 classes). Only BMS's PROTAC drug (BMS-986458) has entered Phase I clinical trials. Therefore, developing novel small molecule degraders that balance druggability and safety, and can rapidly and efficiently degrade BCL6 remains urgent and necessary.

Summary of the Invention

The purpose of this invention is to provide a class of BCL6 protein-targeted degradation chimeras and their uses.

The above-mentioned objective of this invention is achieved through the following technical solution:

A compound with a chemical structure as shown in Formula I, or a pharmaceutically acceptable salt or solvate thereof:

POI-L-E3L

Ⅰ

in:

L is the Linker connecting POI and E3L;

E3L is independently selected from:

in:

_R1 , _R2 , R3 _{, R4} , _R5 , R6, _{R7, R8, R9 , R10} , R11, _R12 , _R13 , _R14 , _{R15 , R16} , R17, _{R18 , R19} , _R20 , _{and R21} are independently selected from -H, halogen, _-C1 to _C3 alkyl, _-C1 to _C3 haloalkyl, and _-C1 to _C3 alkoxy.

R ₂₂ is independently selected from -H, _-C1 to _-C3 alkyl groups;

POIs are selected independently from:

in:

R ₂₃ is independently selected from -H and halogens;

_R24 and _R25 are independently selected from -H and _-C1 to _C3 alkyl groups;

L is selected independently from:

in:

n = 1, 2 or 3.

A compound or a pharmaceutically acceptable salt or solvate thereof showing one of the following chemical structures:

A pharmaceutical composition comprising the above-mentioned compound or a pharmaceutically acceptable salt or solvate thereof.

The above-mentioned compounds or their pharmaceutically acceptable salts or solvates are used in the preparation of BCL6 protein degrading agents.

The above-mentioned compounds or their pharmaceutically acceptable salts or solvates are used in the preparation of medicaments for treating diseases that are treated or alleviated by degrading BCL6 protein.

Preferably, the disease is cancer.

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

The above pharmaceutical composition is used for the preparation of a medicament for treating a disease that is treated or alleviated by degrading BCL6 protein.

Preferably, the disease is cancer.

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

A method of treating a disease, comprising administering to an individual suffering from the disease a therapeutically effective amount of any of the above-mentioned compounds or a pharmaceutically acceptable salt or solvate thereof, or administering a therapeutically effective amount of the above-mentioned pharmaceutical composition; wherein the disease is a disease that is treated or alleviated by degrading BCL6 protein.

Preferably, the disease is cancer.

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

Beneficial effects:

The compounds provided by this invention, or their pharmaceutically acceptable salts or solvates, are protein-targeting degradation chimeras with novel chemical structures. These compounds, or their pharmaceutically acceptable salts or solvates, exhibit excellent degradation activity against BCL6 protein and can be used to prepare BCL6 degrading agents. They have the potential to be developed into drugs for treating or alleviating diseases by degrading BCL6 protein. These diseases include cancers such as Hodgkin lymphoma, B-cell non-Hodgkin lymphoma, T-cell non-Hodgkin lymphoma, NK/T-cell non-Hodgkin lymphoma, and diffuse large B-cell lymphoma.

Detailed Implementation

The following describes the substantive content of the present invention in detail with reference to embodiments, but this does not limit the scope of protection of the present invention.

Synthesis Route 1:

Intermediate 11 was synthesized according to synthetic route 1.

Synthetic Route 1. Reagents and Conditions: (a) 1,2- _{Dibromoethane} , _K₂CO₃ , DMF, rt, 3h; (b) _BBr₃ , DCM, 0℃, 4h; (c) _Cs₂CO₃ , _CH₃CN , 60℃, 1h; (d) _{KNO₃ , H₂SO₄} , ₀ ℃, 4h; (e) Diazonylethyl acetate, DBU, EtOH, DMF, rt, 16h; (f ₎ NaOH, _H₂O , 115℃, 24h; (g) 2-Bromo-N-methylacetamide, _Cs₂CO₃ , DMF, rt, 3h; (h) Fe, _NH₄Cl , EtOH, _H₂O , 80℃, 3h; (i) DIPEA, DMSO, 100℃, 3h.

Intermediate 11 was synthesized according to synthetic route 1.

1-(2-Bromoethyl)-7-methoxyindoline-2,3-dione (2)

1-(2-bromoethyl)-7-methoxyindoline-2,3-dione(2)

1a (5.00 g, 28.22 mmol), 1,2-dibromoethane 1b (10.6 g, 56.45 mmol), and potassium carbonate (7.80 g, 56.45 mmol) were added to a round-bottom flask, followed by 35 mL of DMF solution. The mixture was stirred at room temperature for 3 h, and the reaction was monitored by TLC until complete. 350 mL of water was added and stirred for half an hour. The mixture was then filtered, washed with water, and dried to obtain a red solid intermediate 2 (6.80 g, yield: 84.8%). ^¹H 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). ESI-MS:m/z:[M+H] ⁺ 283.98.

1-(2-Bromoethyl)-7-hydroxyindoline-2,3-dione (3)

1-(2-bromoethyl)-7-hydroxyindoline-2,3-dione(3)

Intermediate 2 (0.30 g, 1.06 mmol) was added to a round-bottom flask containing 6 mL of dichloromethane solution. The flask was in an ice bath for 30 min, followed by the slow addition of boron tribromide (0.79 g, 3.17 mmol). After 1 h, the ice bath was removed, and the reaction was allowed to proceed at room temperature for 2 h. The reaction was monitored by TLC until complete. The reaction was then in an ice bath for half an hour, followed by the slow addition of anhydrous methanol. The addition was stopped when no white fumes were produced. The mixture was concentrated under low pressure, and the residue was purified by column chromatography (elution system: petroleum ether: ethyl acetate = 4:1, v/v) to give intermediate 3 as a red solid (0.16 g, yield: 56.1%). ¹ H NMR (300MHz, DMSO-d ₆ )δ10.48(s,1H),7.17(dd,J＝8.0,1.3Hz,1H),7.09(dd,J＝7.4,1.3Hz,1H),7.03–6.95(m,1H),4.21(t,J＝6.9Hz,2H),3.70(t,J＝6.9Hz,2H). ESI-MS:m/z:[M+H] ⁺ 269.97.

2,3-Dihydro-[1,4]oxazindo[2,3,4-hi]indole-5,6-dione (4)

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

Intermediate 3 (1.00 g, 3.70 mmol) was added to a round-bottom flask, followed by 30 mL of acetonitrile solution, and then cesium carbonate (2.41 g, 7.41 mmol). The mixture was heated at 60 °C for 1 h, and the reaction was monitored by TLC until completion. Heating was removed, and the reaction mixture was allowed to cool to room temperature. The mixture was filtered, washed with ethyl acetate, concentrated, and the residue was purified by column chromatography (elution system: petroleum ether: ethyl acetate = 4:1, v/v) to give intermediate 4 as a red solid (0.57 g, yield: 81.8%). ^¹H 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). ESI-MS: m/z: [M+H] ⁺ 190.04.

2,3-Dihydro-[1,4]oxazindo[2,3,4-hi]indole-5,6-dione (5)

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

Intermediate 4 (2.40 g, 12.69 mmol) was added to a round-bottom flask and stirred in an ice bath for 15 min. Then, 20 mL of concentrated sulfuric acid solution was slowly added, and the mixture was stirred in an ice bath for 0.5 h. Subsequently, potassium nitrate (1.28 g, 12.69 mmol) was slowly added, and the mixture was reacted at room temperature for 6 h. The reaction was monitored by TLC until it was complete. The reaction solution was slowly poured into a stirred ice-water bath, and ethyl acetate was added. The mixture was extracted three times, and the organic phases were combined and concentrated under reduced pressure. The residue was purified by column chromatography (elution system: petroleum ether: ethyl acetate = 4:1, v/v) to give intermediate 5 as an orange-red solid (1.80 g, yield: 60.6%). ^¹H 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] ⁺ 235.03.

6-Hydroxy-9-nitro-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-7-carboxylic acid ethyl ester (6)

Ethyl-6-hydroxy-9-nitro-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-7-carboxylate(6)

Intermediate 5 (1.10 g, 4.70 mmol) was added to a round-bottom flask, followed by a mixed solvent of EtOH:DMF = 20 mL: 4 mL. The mixture was purged with argon, and DBU (107 mg, 0.70 mmol) was quickly added, followed by argon purging. The solution rapidly thickened, changing from orange-red to brownish-yellow (approximately 15 minutes). Then, ethyl diazonate (1.07 g, 9.40 mmol) was quickly added, followed by argon purging. The mixture was stirred at room temperature for 16 h. After the reaction was completed by TLC monitoring, most of the ethanol was removed by concentration under reduced pressure. Dilute hydrochloric acid (16 mL, 1.2 M) was slowly added dropwise using a dropping funnel, followed by 20 mL of water. The mixture was stirred at room temperature for 2 h. Ethyl acetate was added, and the mixture was extracted twice. The organic phases were combined and concentrated under reduced pressure to obtain crude intermediate 6, an orange-yellow oil (1.40 g, yield: 93.0%). ESI-MS: m/z: [M+H] ⁺ 321.06.

6-Hydroxy-9-nitro-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-5-one (7)

6-hydroxy-9-nitro-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-5-one(7)

Intermediate 6 (1.40 g, 4.40 mmol) was added to a round-bottom flask, followed by sodium hydroxide solid (0.35 g, 8.80 mmol), and then 44 mL of water. The mixture was stirred at 115 °C for 24 h, and the reaction was monitored by TLC until completion. The pH was adjusted to 2–3 by slowly adding 1 M dilute hydrochloric acid solution. The mixture was filtered, washed with water, and the filter cake was dried to obtain crude intermediate 7, a yellow solid (0.78 g, yield: 71.9%). ESI-MS: m/z: [M+H] ⁺ 249.05.

N-Methyl-2-(9-nitro-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-6-yl)oxy)acetamide (8)

N-methyl-2-((9-nitro-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-6-yl)oxy)acetamide(8)

Intermediate 7 (1.00 g, 4.03 mmol) was added to a round-bottom flask, followed by 7a (0.74 g, 4.83 mmol) and cesium carbonate (2.63 g, 8.06 mmol), then DMF (15 mL). The mixture was stirred at room temperature for 4 h, and the reaction was monitored by TLC until it was complete. 50 mL of water was added, and the mixture was filtered, washed with water, and the filter cake was dried. Equal volumes of ethyl acetate and water were added to the filtrate for three hydration extractions. The organic layers were combined and concentrated under low pressure to remove the solvent. The residue was combined with the filter cake and purified by column chromatography (elution system: dichloromethane:methanol = 60:1, v/v) to obtain intermediate 8 as a white solid (0.48 g, yield: 37.3%). ¹ H NMR (300MHz, DMSO-d ₆ )δ8.27(d,J＝2.5Hz,1H),7.96(s,1H),7.75(d,J＝2.5Hz,1H),7.54(s,1H),4.6 2(s,2H),4.48(t,J=4.8Hz,2H),4.25(t,J=4.8Hz,2H),2.69(d,J=4.6Hz,3H). ESI-MS:m/z:[M+H] ⁺ 320.08.

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

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

Add 8 (0.40 g, 1.25 mmol) to a round-bottom flask containing anhydrous ethanol:water = 20 mL: 4 mL, add ammonium chloride (0.35 g, 6.26 mmol), and add reduced iron powder (0.34 g, 6.26 mmol) while stirring. Heat the mixture at 80 °C for 3 h, and monitor the reaction for completion by TLC. Remove heating and allow the reaction solution to cool to room temperature. Add 2 mL of ammonia-methanol solution to adjust the pH of the solution to be greater than 7. Filter with diatomaceous earth, wash with dichloromethane:methanol = 20:1 (1000 mL), collect the filtrate, and concentrate under low pressure to obtain a pale yellow solid 9 (0.24 g, yield: 67.0%). ESI-MS: m/z: [M+H] ⁺ 290.11.

2-((9-((2,5-dichloropyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-6-yl)oxy)-N-methylacetamide (11)

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(11)

Intermediate 9 (0.40 g, 1.38 mmol), starting material 10 (0.51 g, 2.77 mmol), and DIPEA (0.54 g, 4.15 mmol) were added to a round-bottom flask, followed by 8 mL of DMSO solution. The mixture was stirred at 100 °C for 3 h, and the reaction was monitored by TLC until complete. 35 mL of water was added and stirred for half an hour. The mixture was then filtered, washed with water, and dried to obtain a pale yellow solid, intermediate 11 (0.43 g, yield: 71.6%). ESI-MS: m/z: [M+H] ⁺ 436.04.

Synthesis Route 2:

Examples 1-3 were synthesized according to synthetic route 2.

Synthetic Route 2: Reagents and Conditions: (a) NaBH(OAc) ₃ , DCE, rt, 3h; (b) _H2 , Pd/C, EtOH, rt, 16h; (c) DIPEA, DMSO, 100℃, 3h; (d) TFA, DCM, rt, 1h; (e) DIPEA, NMP, 170℃, microwave, 3h.

4-((1-((benzyloxy)carbonyl)piperidin-4-yl)methyl)piperazine-1-carboxylic acid tert-butyl ester (21)

tert-butyl 4-((1-((benzyloxy)carbonyl)piperidin-4-yl)methyl)piperazine-1-carboxylate(21)

Synthetic Method 1: Intermediate 20a (151 mg, 0.81 mmol) and 20b (100 mg, 0.40 mmol) were added to a round-bottom flask containing 4 mL of anhydrous 1,2-dichloroethane solution. The mixture was stirred at room temperature for 15 minutes, then NaBH(OAc) ₃ (171 mg, 0.81 mmol) was added, and the mixture was stirred overnight at room temperature. The reaction was monitored by TLC until complete. The reaction solution was concentrated under low pressure and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (elution system: dichloromethane:methanol = 100:1, v/v) to give intermediate 21 as a white solid (201 mg, yield: 59.4%). ¹ H NMR (300MHz, DMSO-d ₆ )δ7.36(qd,J＝6.7,2.6Hz,5H),5.08(s,2H),3.32(dd,J＝12.5,7.6Hz,6H),2.28(t,J＝5.0Hz,4H),2.13(d ,J＝6.8Hz,2H),2.01(s,1H),1.70(d,J＝12.1Hz,3H),1.41(s,9H),1.28–1.16(m,1H).ESI-MS:m/z:[M+H] ^+418.26 .

4-(piperidin-4-ylmethyl)piperazine-1-carboxylic acid tert-butyl ester (22)

tert-butyl 4-(piperidin-4-ylmethyl)piperazine-1-carboxylate(22)

Synthetic Method 2: Intermediate 21 (201 mg, 0.48 mmol) was dissolved in a round-bottom flask containing 5 mL of anhydrous ethanol solution. Palladium on carbon (30 mg, 15% m/m) was added, and the mixture was purged with hydrogen. The mixture was stirred at room temperature for 16 hours, and the reaction was monitored by TLC until complete. The reaction solution was filtered through diatomaceous earth, washed with dichloromethane, and the filtrate was concentrated under low pressure. The residue was purified by column chromatography (elution system: dichloromethane:methanol = 100:1, v/v) to obtain intermediate 22, a white oily substance (108 mg, yield: 79.2%). ESI-MS: m/z: [M+H] ⁺ 283.23.

4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-4-yl)piperidin-4-yl)methyl)piperazine-1-carboxylic acid tert-butyl ester (24a)

tert-butyl-4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)piperidin-4-yl)methyl)piperazine-1-carboxylate(24a)

Synthetic Method 3: Intermediate 23a (400 mg, 1.45 mmol) and 22 (534 mg, 1.88 mmol) were added to a round-bottom flask containing 6 mL of DMSO solution, followed by the addition of DIPEA (374 mg, 2.90 mmol). The mixture was stirred at 100 °C for 3 hours, and the reaction was monitored by TLC until complete. After the reaction cooled to room temperature, the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (elution system: dichloromethane:methanol = 100:1, v/v) to obtain intermediate 24a as a yellow solid (690 mg, yield: 88.3%). ¹ H NMR (300MHz, DMSO-d ₆ )δ11.13(s,1H),7.69(dd,J＝8.5,7.1Hz,1H),7.34(dd,J＝8.0,4.1Hz,2H),5.11(dd,J＝13.1,5.7Hz,1H),3.70(d,J＝11.7Hz,1H),2.88(s,2H),2. 56(s,6H),2.32(s,3H),2.21(d,J＝6.8Hz,1H),2.03(d,J＝8.9Hz,1H),1. 83(d,J＝13.2Hz,2H),1.41(s,9H),1.37–1.23(m,3H).ESI-MS:m/z:[M+H] ^+540.27 .

4-((1-(2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)piperidin-4-yl)methyl)piperazine-1-carboxylic acid tert-butyl ester (24b)

tert-butyl-4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methyl)piperazine-1-carboxylate(24b)

Following general synthesis method 3, using 23b (400 mg, 1.45 mmol) as the reactant, the target product, yellow solid 24b (690 mg, yield: 88.3%), was obtained. ¹ H NMR (300MHz, DMSO-d ₆ )δ7.68(dd,J＝8.5,7.1Hz,1H),7.33(dd,J＝7.8,5.4Hz,2H),5.09(dd,J＝12 .7,5.3Hz,1H),2.88(t,J＝12.4Hz,3H),2.62(s,1H),2.56(s,1H),2.43(s, 3H),2.23(d,J＝6.8Hz,2H),2.02(d,J＝11.4Hz,1H),1.81(t,J＝16.9Hz,3H) ,1.42–1.30(m,2H),1.25(s,9H),1.24(d,J＝4.4Hz,1H).ESI-MS:m/z:[M+H] ⁺ 540.27.

4-((1-(2-(2-(2,6-dioxopiperidin-3-yl)-6-fluoro-1,3-dioxoisoindoline-5-yl)piperidin-4-yl)methyl)piperazine-1-carboxylic acid tert-butyl ester (24c)

tert-butyl-4-((1-(2-(2,6-dioxopiperidin-3-yl)-6-fluoro-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methyl)piperazine-1-carboxylate(24c)

Following general synthetic method 3, using 23c (400 mg, 1.36 mmol) as the reactant, the target product, a yellow solid 24c (670 mg, yield: 91.3%), was obtained. ^¹H NMR (300 MHz, DMSO- _d6) )δ11.15(s,1H),7.73(d,J＝11.4Hz,1H),7.45(d,J＝7.4Hz,1H),5.12(dd,J＝12.9 ,5.3Hz,1H),3.61(d,J＝12.0Hz,2H),2.88(dd,J＝15.4,7.6Hz,3H),2.65–2.53(m ,2H),2.30(d,J＝6.2Hz,4H),2.19(d,J＝6.8Hz,2H),2.07–2.00(m,1H),1.83(d,J ＝13.3Hz,2H),1.41(d,J＝4.4Hz,10H),1.26(d,J＝9.7Hz,2H).ESI-MS:m/z:[M+H] ⁺ 558.26.

2-(2,6-dioxopiperidin-3-yl)-4-(4-(piperazin-1-ylmethyl)piperidin-1-yl)isoindoline-1,3-dione (25a)

2-(2,6-dioxopiperidin-3-yl)-4-(4-(piperazin-1-ylmethyl)piperidin-1-yl)isoindoline-1,3-dione(25a)

Synthetic Method 4: Intermediate 24a (300 mg, 0.56 mmol) was added to a round-bottom flask containing 5 mL of dichloromethane solution, followed by 1.5 mL of trifluoroacetic acid. The mixture was stirred at room temperature for 1 hour, and the reaction was monitored by TLC until complete. The solution was concentrated under reduced pressure to obtain intermediate 25a, a yellow oil (230 mg, yield: 94.1%). ESI-MS: m/z: [M+H] ⁺ 440.22.

2-(2,6-dioxopiperidin-3-yl)-5-(4-(piperazin-1-ylmethyl)piperidin-1-yl)isoindoline-1,3-dione (25b)

2-(2,6-dioxopiperidin-3-yl)-5-(4-(piperazin-1-ylmethyl)piperidin-1-yl)isoindoline-1,3-dione(25b)

Following general synthesis method 4, using 24b (300 mg, 0.56 mmol) as the reactant, the target product, a yellow oily substance 25b (230 mg, yield: 94.1%), was obtained. ESI-MS: m/z: [M+H] ⁺ 440.22.

2-(2,6-dioxopiperidin-3-yl)-5-fluoro-6-(4-(piperazin-1-ylmethyl)piperidin-1-yl)isoindoline-1,3-dione (25c)

2-(2,6-dioxopiperidin-3-yl)-5-fluoro-6-(4-(piperazin-1-ylmethyl)piperidin-1-yl)isoindoline-1,3-dione(25c)

Following general synthesis method 4, using 24c (300 mg, 0.54 mmol) as the reactant, the target product, a yellow oily substance 25c (210 mg, yield: 85.3%), was obtained. ESI-MS: m/z: [M+H] ⁺ 458.21.

Example 1

2-((9-((5-chloro-2-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-4-yl)piperidin-4-yl)methyl)piperazin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-6-yl)oxy)-N-methylacetamide (Example 1)

2-((9-((5-chloro-2-(4-((1-(2-(2,6-dioxopiperidin-3-yl))-1,3-dioxoisoindolin-4-yl)piperidin-4-yl)methyl)piperazin- 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 (Example 1)

Synthetic Method 5: Intermediate 25a (121 mg, 0.28 mmol) and 11 (60 mg, 0.14 mmol) were added to a round-bottom flask containing 5 mL of NMP solution, followed by the addition of DIPEA (71 mg, 0.55 mmol). The mixture was microwaved at 170 °C for 3 hours, and the reaction was monitored by TLC until complete. After cooling to room temperature, the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by preparative chromatography (elution system: dichloromethane:methanol = 20:1, v/v) to obtain the target product of Example 1, as a yellow solid (18 mg, yield: 15.6%). ^¹H NMR (300 MHz, DMSO- _d6) was performed. )δ11.12(s,1H),8.79(s,1H),8.03(d,J＝13.0Hz,2H),7.75–7.54(m,2H),7.32(d,J＝ 10.8Hz,3H),7.10(s,1H),5.09(dd,J＝13.6,5.2Hz,1H),4.60(s,2H),4.38(s,2H),4 .16(s,2H),3.66(s,6H),2.84(d,J＝12.9Hz,3H),2.66(d,J＝4.4Hz,6H),2.40(s,3H) ,2.22(s,2H),2.01(d,J＝9.9Hz,1H),1.83(d,J＝13.1Hz,3H),1.27(d,J＝31.9Hz,2H). ¹³ C NMR (75MHz, DMSO-d ₆ )δ173.38,170.60,167.93,167.61,159.58,155.68,155.15,147.59,142.95,136.25,134.43,134.15,124.42,120.38,118.91,116.78,1 14.88,113.51,112.48,109.28,102.71,68.34,64.20,53.42,51.49,49.23,44.34,32.53,31.47,30.99,25.93,22.57.HRMS(ESI):calcd for C ₄₁ H ₄₃ ClN ₁₀ O ₈ [M+H] ⁺ 839.2954, found 839.3023. Purity: 96.63% by HPLC (MeOH/H ₂ O = 80:20, t _R = 4.042 min).

Example 2

2-((9-((5-chloro-2-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)piperidin-4-yl)methyl)piperazin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-6-yl)oxy)-N-methylacetamide (Example 2)

2-((9-((5-chloro-2-(4-((1-(2-(2,6-dioxopiperidin-3-yl))-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methyl)piperazin- 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 (Example 2)

Following general synthesis method 5, using 25b (121 mg, 0.28 mmol) as the reactant, the target product, a yellow solid, was obtained (Example 2, 18 mg, yield: 15.6%). ^¹H NMR (300 MHz, DMSO- _d₆ ) δ 11.10 (s, 1H), 8.78 (s, 1H), 8.07 (s, 1H), 7.98 (d, J = 4.8 Hz, 1H), 7.71–7.58 (m, 2H), 7.41–7.30 (m, 2H), 7.24 (d, J = 8.6 Hz, 1H), 7.12 (s, 1H), 5.08 (dd, J = 12.7, 5.4 Hz, 1H), 4.61 (s, 2H), 4.40 (t, J = 4.7 Hz, 2H). H),4.19(d,J＝4.6Hz,2H),4.06(d,J＝12.9Hz,2H),3.66(s,4H),2.94(dt,J＝24.8,12.4Hz,3H),2.70– 2.55(m,4H),2.41(s,3H),2.19(s,2H),2.10–1.99(m,1H),1.84(d,J=13.2Hz,2H),1.31–1.12(m,3H). ¹³ C NMR (75MHz, DMSO-d ₆ )δ173.31,170.62,168.14,167.94,167.46,159.61,155.71,155.50,155.15 ,155.07,147.63,142.96,134.54,134.45,125.50,120.38,118.96,118.07, 117.84,113.69,112.47,109.30,108.23,102.73,68.45,64.22,53.40,49.2 4,47.76,44.37,40.74,32.89,31.50,30.13,25.92,22.70.HRMS(ESI):calcd for C ₄₁ H ₄₃ ClN ₁₀ O ₈ [M+H] ⁺ 839.2954, found 839.3026. Purity: 98.43% by HPLC (MeOH/H ₂ O=80:20, t _R =4.012min).

Example 3

2-((9-((5-chloro-2-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-6-fluoro-1,3-dioxoisoindoline-5-yl)piperidin-4-yl)methyl)piperazin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-6-yl)oxy)-N-methylacetamide (Example 3)

2-((9-((5-chloro-2-(4-((1-(2-(2,6-dioxopiperidin-3-yl))-6-fluoro-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methyl)piper azin-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 (Example 3)

Following general synthesis method 5, using 25c (126 mg, 0.28 mmol) as the reactant, the target product was obtained as a yellow solid (Example 3, 21 mg, yield: 17.8%). ^¹H NMR (300 MHz, DMSO- _d₆ ) δ 11.13 (s, 1H), 8.80 (s, 1H), 8.07 (s, 1H), 7.99 (d, J = 4.8 Hz, 1H), 7.72 (d, J = 11.4 Hz, 1H), 7.61 (d, J = 2.2 Hz, 1H), 7.45 (d, J = 7.4 Hz, 1H), 7.36 (d, J = 2.1 Hz, 1H), 7.13 (s, 1H), 5.11 (dd, J = 13.0, 5.3 Hz, 1H), 4.61 (s, 2.8 Hz, 1H). H),4.40(s,2H),4.18(s,2H),3.63(d,J＝14.9Hz,6H),2.91(t,J＝12.3Hz,3H),2.67(d,J＝4.6Hz,3H),2.61 (d,J＝14.8Hz,2H),2.42(s,3H),2.23(s,2H),2.03(d,J＝7.8Hz,1H),1.86(d,J＝12.6Hz,2H),1.25(s,4H). ¹³ C NMR (75MHz, DMSO-d ₆ )δ173.27,170.43,167.95,167.21,166.72,155.73,155.16,155.07,147.63,142.96,134.43,129.30,120.39,118.98,1 13.68,112.51,109.32,102.79,68.44,64.22,53.34,50.40,49.54,44.24,31.46,30.71,25.93,22.58.HRMS(ESI):calcd for C ₄₁ H ₄₂ ClFN ₁₀ O ₈ [M+H] ⁺ 857.2860, found 857.2930. Purity: 95.79% by HPLC (MeOH/ _H₂O = 80:20, _tR = 4.172 min).

Synthesis Route 3:

Example 4 was synthesized according to synthesis route 3.

Synthetic Route 3: Reagents and Conditions: (a) Pd(OAc) _₂ , Xantphos, _Cs₂CO₃ , _dioxane , 110℃, 10h; (b) LiOH, MeOH, _H₂O , 40℃, 10h; (c) EDCI, HOBt, DIPEA, DMF, 25℃, 10h; (d) TFA, DCM, rt, 1h; (e) DIPEA, NMP, 170℃, microwave, 3h.

4-((1-(3-fluoro-4-(methoxycarbonyl)phenyl)piperidin-4-yl)methyl)piperazine-1-carboxylic acid tert-butyl ester (26)

tert-butyl-4-((1-(3-fluoro-4-(methoxycarbonyl)phenyl)piperidin-4-yl)methyl)piperazine-1-carboxylate(26)

Synthetic Method 6: Intermediate 22a (360 mg, 1.54 mmol) and 22 (482 mg, 1.70 mmol) were added to a sealed tube containing 10 mL of anhydrous dioxane solution. Cesium carbonate (1.01 g, 3.09 mmol), palladium acetate (17 mg, 0.08 mmol), and XantPhos (89 mg, 0.15 mmol) were then added sequentially. The mixture was purged with argon and reacted at 110 °C for 16 hours. The reaction was monitored by TLC until complete. After cooling to room temperature, the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by preparative chromatography (elution system: dichloromethane:methanol = 100:1, v/v) to give intermediate 26 as a white solid (390 mg, yield: 58.0%). ESI-MS: m/z: [M+H] ⁺ 436.25.

4-(4-((4-(tert-Butoxycarbonyl)piperazin-1-yl)methyl)piperidin-1-yl)-2-fluorobenzoic acid (27)

4-(4-((4-(tert-butoxycarbonyl)piperazin-1-yl)methyl)piperidin-1-yl)-2-fluorobenzoic acid(27)

Synthetic Method 7: Intermediate 26 (200 mg, 0.46 mmol) was added to a round-bottom flask containing a methanol:water mixture of 8 mL:2 mL, followed by lithium hydroxide (55 mg, 2.30 mmol). The reaction was carried out at 40 °C for 10 hours, and the reaction was monitored by TLC until complete. After the reaction cooled to room temperature, dilute hydrochloric acid was added to adjust the pH of the solution to 5–6. The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (elution system: dichloromethane:methanol = 100:1, v/v) to obtain intermediate 27 as a white solid (180 mg, yield: 93.0%). ESI-MS: m/z: [M+H] ⁺ 421.24.

4-((1-(4-((2,6-dioxopiperidin-3-yl)carbamoyl)-3-fluorophenyl)piperidin-4-yl)methyl)piperazine-1-carboxylic acid tert-butyl ester (28)

tert-butyl-4-((1-(4-((2,6-dioxopiperidin-3-yl)carbamoyl)-3-fluorophenyl)piperidin-4-yl)methyl)piperazine-1-carboxylate(28)

Synthetic Method 8: Intermediate 27 (180 mg, 0.43 mmol) was added to a round-bottom flask containing 5 mL of LDM solution, followed by DIPEA (276 mg, 2.14 mmol), EDCI (123 mg, 0.64 mmol), and HOBt (87 mg, 0.64 mmol). The mixture was stirred at room temperature for half an hour, then 27a (83 mg, 0.51 mmol) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until complete. The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (elution system: dichloromethane:methanol = 50:1, v/v) to give intermediate 28 as a white solid (160 mg, yield: 70.5%). ESI-MS: m/z: [M+H] ⁺ 532.28.

N-(2,6-dioxopiperidin-3-yl)-2-fluoro-4-(4-(piperazin-1-ylmethyl)piperidin-1-yl)benzamide (29)

N-(2,6-dioxopiperidin-3-yl)-2-fluoro-4-(4-(piperazin-1-ylmethyl)piperidin-1-yl)benzamide(29)

Following general synthesis method 4, using 28 (200 mg, 0.38 mmol) as the reactant, the target product, a white oily substance 29 (130 mg, yield: 80.1%), was obtained. ESI-MS: m/z: [M+H] ⁺ 432.23.

Example 4

4-(4-((4-(5-chloro-4-((6-(2-(methylamino)-2-oxoethoxy)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-9-yl)amino)pyrimidin-2-yl)piperazin-1-yl)methyl)piperidin-1-yl)-N-(2,6-dioxopiridin-3-yl)-2-fluorobenzamide (Example 4)

4-(4-((4-(5-chloro-4-((6-(2-(methylamino)-2-oxoethoxy))-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-9-yl )amino)pyrimidin-2-yl)piperazin-1-yl)methyl)piperidin-1-yl)-N-(2,6-dioxopiperidin-3-yl)-2-fluorobenzamide (Example 4)

Following general synthesis method 5, using 29 (119 mg, 0.28 mmol) as the reactant, the target product was obtained as a yellow solid (Example 4, 25 mg, yield: 21.9%). ^¹H NMR (300 MHz, DMSO-d6 ₎ )δ10.87(s,1H),8.79(s,1H),8.07(s,1H),8.00(q,J＝4.1,3.7Hz,1H),7.92(t,J＝7.8Hz,1H),7.66(t,J＝8.9Hz,1H),7.59(d, J＝2.3Hz,1H),7.40(d,J＝2.2Hz,1H),7.12(s,1H),6.43(dd,J＝9.1,2.2Hz,1H),6.35(d,J＝15.1Hz,1H),4.74(dt,J＝12.4,6.4H z,1H),4.60(s,2H),4.51(d,J＝12.0Hz,2H),4.41(d,J＝4.4Hz,2H),4.21–4.16(m,2H),4.03(d,J＝12.9Hz,1H),3.17(s,2H),2. 97(t,J＝11.9Hz,2H),2.67(d,J＝4.6Hz,3H),2.15(d,J＝11.7Hz,2H),1.86(m,1H),1.76(m,2H),1.32(m,1H).HRMS(ESI):calcd for C ₄₀ H ₄₄ ClFN ₁₀ O ₇ [M+H] ⁺ 831.3067, found 831.3149. Purity: 96.19% by HPLC (MeOH/H ₂ O = 80:20, t _R = 4.022min).

Synthesis Route 4:

Examples 5 and 6 were synthesized according to synthesis route 4.

Synthetic Route 4: Reagents and Conditions: (a) NaBH(OAc) ₃ , DCE, rt, 12h; (b) TFA, DCM, rt, 3h; (c) DIPEA, DMSO, 100℃, 3h; (d) _H2 , Pd/C, EtOH, rt, 16h; (e) DIPEA, NMP, microwave, 170℃, 3h.

4-((4-(2-((tert-Butoxycarbonyl)amino)ethyl)piperidin-1-yl)methyl)piperidin-1-carboxylic acid benzyl ester (31a)

benzyl-4-((4-(2-((tert-butoxycarbonyl)amino)ethyl)piperidin-1-yl)methyl)piperidine-1-carboxylate(31a)

Following general synthetic method 1, using intermediate 30a (400 mg, 1.75 mmol) as the reactant, the target product, white solid 31a (610 mg, yield: 75.8%), was obtained. ESI-MS: m/z: [M+H] ⁺ 460.31.

4-((4-((tert-Butoxycarbonyl)amino)piperidin-1-yl)methyl)piperidin-1-carboxylic acid benzyl ester (31b)

benzyl-4-((4-((tert-butoxycarbonyl)amino)piperidin-1-yl)methyl)piperidine-1-carboxylate(31b)

Following general synthetic method 1, using intermediate 30b (400 mg, 2.00 mmol) as the reactant, the target product, white solid 31b (610 mg, yield: 75.8%), was obtained. ESI-MS: m/z: [M+H] ⁺ 432.28.

4-((4-(2-aminoethyl)piperidin-1-yl)methyl)piperidin-1-carboxylic acid benzyl ester (32a)

benzyl 4-((4-(2-aminoethyl)piperidin-1-yl)methyl)piperidine-1-carboxylate(32a)

Following general synthetic method 4, using intermediate 31a (400 mg, 0.87 mmol) as the reactant, the target product, a white oily substance 32a (290 mg, yield: 92.7%), was obtained. ESI-MS: m/z: [M+H] ⁺ 360.26.

4-((4-aminopiperidin-1-yl)methyl)piperidin-1-carboxylic acid benzyl ester (32b)

benzyl 4-((4-aminopiperidin-1-yl)methyl)piperidine-1-carboxylate(32b)

Following general synthetic method 4, using intermediate 31b (400 mg, 0.87 mmol) as the reactant, the target product, a white oily substance 32b (270 mg, yield: 93.6%), was obtained. ESI-MS: m/z: [M+H] ⁺ 332.23.

4-((4-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-4-yl)amino)ethyl)piperidin-1-yl)methyl)piperidin-1-carboxylic acid benzyl ester (33a)

Benzyl-4-((4-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)ethyl)piperidin-1-yl)methyl)piperidine-1-carboxylate(33a)

Following general synthesis method 3, using 23a (400 mg, 1.45 mmol) as the reactant, the target product, a yellow solid 33a (840 mg, yield: 94.2%), was obtained. ESI-MS: m/z: [M+H] ⁺ 616.31.

4-((4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-4-yl)amino)piperidin-1-yl)methyl)piperidin-1-carboxylic acid benzyl ester (33b)

Benzyl-4-((4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)piperidin-1-yl)methyl)piperidine-1-carboxylate(33b)

Following general synthesis method 3, using 23a (400 mg, 1.45 mmol) as the reactant, the target product, a yellow solid 33b (810 mg, yield: 95.2%), was obtained. ESI-MS: m/z: [M+H] ⁺ 588.27.

2-(2,6-dioxopiperidin-3-yl)-4-((2-(1-(piperidin-4-ylmethyl)piperidin-4-yl)ethyl)amino)isoindoline-1,3-dione (34a)

2-(2,6-dioxopiperidin-3-yl)-4-((2-(1-(piperidin-4-ylmethyl)piperidin-4-yl)ethyl)amino)isoindoline-1,3-dione(34a)

Following general synthesis method 2, using 33a (300 mg, 0.49 mmol) as the reactant, the target product, a yellow oil 34a (190 mg, yield: 81.0%), was obtained. ESI-MS: m/z: [M+H] ⁺ 482.27.

2-(2,6-dioxopiperidin-3-yl)-4-((1-(piperidin-4-ylmethyl)piperidin-4-yl)amino)isoindoline-1,3-dione (34b)

2-(2,6-dioxopiperidin-3-yl)-4-((1-(piperidin-4-ylmethyl)piperidin-4-yl)amino)isoindoline-1,3-dione(34b)

Following general synthesis method 2, using 33b (300 mg, 0.49 mmol) as the reactant, the target product, a yellow oily substance 34b (180 mg, yield: 76.7%), was obtained. ESI-MS: m/z: [M+H] ⁺ 454.24.

Example 5

2-((9-((5-chloro-2-(4-((4-(2-(2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-4-yl)amino)ethyl)piperidin-1-yl)methyl)piperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-6-yl)oxy)-N-methylacetamide (Example 5)

2-(((9-((5-chloro-2-(4-((4-(2-((2-(2,6-dioxopiperidin-3-yl))-1,3-dioxoisoindolin-4-yl)amino)ethyl)piperidin-1-yl)methyl)pi peridin-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 (Example 5)

Following general synthesis method 5, using 34a (132 mg, 0.28 mmol) as the reactant, the target product, a yellow solid, was obtained in Example 5 (18 mg, yield: 9.1%). ^¹H NMR (300 MHz, DMSO- _d₆ ) δ 11.17 (s, 1H), 8.77 (s, 1H), 8.03 (d, J = 11.6 Hz, 2H), 7.64–7.55 (m, 2H), 7.41 (d, J = 2.2 Hz, 1H), 7.13–7.09 (m, 2H), 7.05 (d, J = 7.1 Hz, 1H), 6.54 (s, 1H), 5.07 (dd, J = 12.8, 5.4 Hz, 1H), 4.61 (s, 2H), 4.50 (d, J = 11.6 Hz, 2H). 12.5Hz,2H),4.40(d,J＝5.4Hz,2H),4.20(d,J＝4.7Hz,2H),2.89(d,J＝14.4Hz,5H),2.68(d,J＝4.6Hz,5 H),2.57(s,4H),2.09–1.97(m,3H),1.73(s,2H),1.54(s,2H),1.26(d,J＝4.4Hz,8H).HRMS(ESI):calcd for C ₄₄ H ₄₉ ClN ₁₀ O ₈ [M+H] ⁺ 881.3423, found 881.3478. Purity: 97.53% by HPLC (MeOH/H ₂ O = 80:20, t _R = 3.998 min).

Example 6

2-((9-((5-chloro-2-(4-((4-((2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-4-yl)amino)piperidin-1-yl)methyl)piperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-6-yl)oxy)-N-methylacetamide (Example 6)

2-((9-((5-chloro-2-(4-((4-((2-(2,6-dioxopiperidin-3-yl))-1,3-dioxoisoindolin-4-yl)amino)piperidin-1-yl)methyl)piperi din-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 (Example 6)

Following general synthesis method 5, using 34b (125 mg, 0.28 mmol) as the reactant, the target product was obtained as a yellow solid (Example 6, 13 mg, yield: 11.1%). ^¹H NMR (300 MHz, DMSO-d6 ₎ )δ11.15(s,1H),9.00(s,1H),8.28(d,J＝10.7Hz,2H),7.89–7.81(m,2H),7.67(d,J＝2.2Hz,1H),7.44(d,J＝8.6Hz,1H),7.36(s ,1H),7.31(d,J＝7.0Hz,1H),6.52(d,J＝8.0Hz,1H),5.33(dd,J＝12.9,5.4Hz,1H),4.86(s,2H),4.76(d,J＝12.7Hz,2H),4.65(d ,J＝5.3Hz,2H),4.45(d,J＝4.8Hz,2H),3.12(t,J＝11.5Hz,3H),2.94(d,J＝4.6Hz,3H),2.88(s,1H),2.81(s,1H),2.41(s,3H),2 .23(dd,J＝17.8,9.7Hz,3H),2.01(d,J＝12.6Hz,2H),1.76(s,2H),1.59–1.47(m,5H),1.29(d,J＝12.3Hz,2H).HRMS(ESI):calcd for C ₄₂ H ₄₅ ClN ₁₀ O ₈ [M+H] ⁺ 853.3110, found 853.3174. Purity: 96.13% by HPLC (MeOH/H ₂ O=80:20, t _R =4.018min).

Synthesis Route 5:

Examples 7-9 were synthesized according to synthetic route 5.

Synthetic Route 5: Reagents and Conditions: (a) NaBH(OAc) ₃ , DCE, rt, 3h; (b) _H2 , Pd/C, EtOH, rt, 16h; (c) DIPEA, DMSO, 100℃, 3h; (d) TFA, DCM, rt, 1h; (e) DIPEA, DMSO, 100℃, 10h; (f) DIPEA, EDCI, HOBt, DMF, rt, 12h.

1-(5-chloro-4-((6-(2-(methylamino)-2-oxoethoxy)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-9-yl)amino)pyrimidin-2-yl)piperidine-4-carboxylic acid (35)

1-(5-chloro-4-((6-(2-(methylamino)-2-oxoethoxy)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-9-yl)amino)pyrimidin-2-yl)piperidine-4-carboxylic acid(35)

Intermediate 11 (300 mg, 0.69 mmol), starting material 12i (178 mg, 1.39 mmol), and DIPEA (151 mg, 1.17 mmol) were added to a round-bottom flask, followed by 8 mL of DMSO solution. The mixture was stirred at 100 °C for 10 h, and the reaction was monitored by TLC until complete. The pH of the solution was adjusted to 2–5 by slow dropwise addition of dilute hydrochloric acid, and then 50 mL of water was added and stirred for 10 minutes. The mixture was filtered, washed with water, and dried to obtain a yellow solid 35 (310 mg, yield: 85.2%). ESI-MS: m/z: [M+H] ⁺ 259.15.

4-(1-((benzyloxy)carbonyl)piperidin-4-yl)piperazine-1-carboxylic acid tert-butyl ester (21c)

tert-butyl 4-(1-((benzyloxy)carbonyl)piperidin-4-yl)piperazine-1-carboxylate(21c)

Following general synthetic method 1, using intermediate 20c (400 mg, 1.71 mmol) as the reactant, the target product, a white oily substance 21c (410 mg, yield: 59.2%), was obtained. ESI-MS: m/z: [M+H] ⁺ 404.25.

4-((1-((benzyloxy)carbonyl)azacyclobutane-3-yl)methyl)piperazine-1-carboxylic acid tert-butyl ester (21d)

tert-butyl 4-((1-((benzyloxy)carbonyl)azetidin-3-yl)methyl)piperazine-1-carboxylate(21d)

Following general synthetic method 1, using intermediate 20d (400 mg, 1.82 mmol) as the reactant, the target product, a white oily substance 21d (440 mg, yield: 61.9%), was obtained. ESI-MS: m/z: [M+H] ⁺ 389.23.

4-(piperidin-4-yl)piperazine-1-carboxylic acid tert-butyl ester (22c)

tert-butyl 4-(piperidin-4-yl)piperazine-1-carboxylate(22c)

Following general synthesis method 2, using 21c (300 mg, 0.74 mmol) as the reactant, the target product, a colorless oil 22c (170 mg, yield: 84.9%), was obtained. ESI-MS: m/z: [M+H] ⁺ 270.21.

4-(azacyclobutane-3-ylmethyl)piperazine-1-carboxylic acid tert-butyl ester (22d)

tert-butyl 4-(azetidin-3-ylmethyl)piperazine-1-carboxylate(22d)

Following general synthesis method 2, using 21d (300 mg, 0.77 mmol) as the reactant, the target product, a colorless oil 22d (180 mg, yield: 91.5%), was obtained. ESI-MS: m/z: [M+H] ⁺ 256.19.

4-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)piperidin-4-yl)piperazine-1-carboxylic acid tert-butyl ester (24c)

tert-butyl-4-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)piperazine-1-carboxylate(24c)

Following general synthesis method 3, using 23b (400 mg, 1.45 mmol) as the reactant, the target product, yellow solid 24c (590 mg, yield: 77.5%), was obtained. ¹ H NMR (300MHz, DMSO-d ₆ )δ7.67(d,J＝8.5Hz,1H),7.33(d,J＝2.2Hz,1H),7.25(dd,J＝8.7,2.3Hz,1H ),5.08(dd,J＝12.6,5.3Hz,1H),4.08(d,J＝13.2Hz,2H),2.92(dt,J＝25.2,9 .1Hz,3H),2.63(s,1H),2.56(d,J＝3.8Hz,2H),2.45(t,J＝4.8Hz,4H),2.03( d,J＝12.1Hz,1H),1.84(d,J＝12.4Hz,2H),1.40(s,11H).ESI-MS:m/z:[M+H] ⁺ 526.26.

4-((1-(2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)azacyclobutane-3-yl)methyl)piperazine-1-carboxylic acid tert-butyl ester (24d)

tert-butyl-4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)azetidin-3-yl)methyl)piperazine-1-carboxylate(24d)

Following general synthesis method 3, using 23b (400 mg, 1.45 mmol) as the reactant, the target product, a yellow solid 24d (610 mg, yield: 82.3%), was obtained. ESI-MS: m/z: [M+H] ⁺ 526.26.

2-(2,6-dioxopiperidin-3-yl)-5-(4-(piperazin-1-yl)piperidin-1-yl)isoindoline-1,3-dione (25c)

2-(2,6-dioxopiperidin-3-yl)-5-(4-(piperazin-1-yl)piperidin-1-yl)isoindoline-1,3-dione(25c)

Following general synthesis method 4, using 24c (300 mg, 0.57 mmol) as the reactant, the target product, a yellow oily substance 25c (205 mg, yield: 84.4%), was obtained. ESI-MS: m/z: [M+H] ⁺ 426.21.

2-(2,6-dioxopiperidin-3-yl)-5-(3-(piperazin-1-ylmethyl)azacyclobutane-1-yl)isoindoline-1,3-dione (25d)

2-(2,6-dioxopiperidin-3-yl)-5-(3-(piperazin-1-ylmethyl)azetidin-1-yl)isoindoline-1,3-dione(25d)

Following general synthesis method 4, using 24d (300 mg, 0.56 mmol) as the reactant, the target product, a yellow oily substance 25d (195 mg, yield: 85.8%), was obtained. ESI-MS: m/z: [M+H] ⁺ 412.19.

Example 7

2-((9-((5-chloro-2-(4-(4-((1-(2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)piperidin-4-yl)methyl)piperazine-1-carbonyl)piperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-6-yl)oxy)-N-methylacetamide (Example 7)

2-((9-((5-chloro-2-(4-(4-((1-(2-(2,6-dioxopiperidin-3-yl))-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)methyl)piperazine-1-carbon yl)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 (Example 7)

Synthetic Method 9: Intermediate 35 (60 mg, 0.11 mmol) was added to a round-bottom flask containing 5 mL of LDMF solution, followed by DIPEA (71 mg, 0.55 mmol), EDCI (32 mg, 0.16 mmol), and HOBt (23 mg, 0.16 mmol). The mixture was stirred at room temperature for half an hour, then 25b (100 mg, 0.23 mmol) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until complete. The mixture was extracted three times with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by preparative chromatography (elution system: dichloromethane:methanol = 15:1, v/v) to obtain the target product of Example 7, a yellow solid (16 mg, yield: 14.8%). ^¹H NMR (300 MHz, DMSO- _d6) was performed. )δ11.10(s,1H),8.78(s,1H),8.07(s,1H),7.99(d,J＝4.8Hz,1H),7.66(s,1H),7.41(s,1H),7.33(s,2H), 7.25(d,J＝8.7Hz,1H),7.12(s,1H),5.09(d,J＝7.3Hz,1H),4.60(s,2H),4.54–4.46(m,2H),4.40(s,2H),4 .19(s,2H),4.05(s,2H),3.57(s,2H),3.46(s,2H),3.01–2.91(m,7H),2.70–2.59(m,5H),2.18(s,2H),2. 07–2.00(m,2H),1.82(d,J＝12.7Hz,4H),1.67(d,J＝12.8Hz,3H),1.50(d,J＝12.3Hz,2H).HRMS(ESI):calcd for C ₄₇ H ₅₂ ClN ₁₁ O ₉ [M+H] ⁺ 950.3638,found 950.3706. Purity: 94.88% by HPLC (MeOH/H ₂ O = 80:20, t _R = 3.903 min).

Example 8

2-((9-((5-chloro-2-(4-(4-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)piperidin-4-yl)piperazin-1-carbonyl)piperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-6-yl)oxy)-N-methylacetamide (Example 8)

2-((9-((5-chloro-2-(4-(4-(1-(2-(2,6-dioxopiperidin-3-yl))-1,3-dioxoisoindolin-5-yl)piperidin-4-yl)piperazine-1-carbonyl)p iperidin-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 (Example 8)

Following general synthesis method 9, using 35 (60 mg, 0.11 mmol) as the reactant, the target product was obtained as a yellow solid (Example 8, 13 mg, yield: 12.2%). ^¹H NMR (300 MHz, DMSO-d6 ₎ )δ11.11(s,1H),8.78(s,1H),8.07(s,1H),7.98(d,J＝5.0Hz,1H),7.68(d,J＝8.5Hz,1H),7.59(d,J＝2.2Hz,1H),7.40(d,J＝2.2Hz,1H),7.35(d ,J＝2.2Hz,1H),7.27(dd,J＝8.7,2.2Hz,1H),7.12(s,1H),5.09(dd,J＝12.8,5.3Hz,1H),4.60(s,2H),4.50(d,J＝12.7Hz,2H),4.40(t,J＝4.7Hz ,2H),4.19(t,J＝4.7Hz,2H),4.10(d,J＝12.7Hz,2H),3.55(s,2H),3.44(s,2H),3.03–2.89(m,6H),2.67(d,J＝4.6Hz,3H),2.63(s,1H),2.57(s ,2H),2.45(s,2H),2.03(d,J＝11.9Hz,1H),1.85(d,J＝12.1Hz,2H),1.66(d,J＝12.5Hz,2H),1.59–1.38(m,5H),1.26(s,1H).HRMS(ESI):calcd for C ₄₆ H ₅₀ ClN ₁₁ O ₉ [M+H] ⁺ 936.3481,found 936.3531. Purity: 96.43% by HPLC (MeOH/H ₂ O = 80:20, t _R = 3.993 min).

Example 9

2-((9-((5-chloro-2-(4-(4-(((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)azacyclobutane-3-yl)methyl)piperazine-1-carbonyl)piperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-6-yl)oxy)-N-methylacetamide (Example 9)

2-((9-((5-chloro-2-(4-(4-((1-(2-(2,6-dioxopiperidin-3-yl))-1,3-dioxoisoindolin-5-yl)azetidin-3-yl)methyl)piperazine-1-carbony l)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 (Example 9)

Following general synthesis method 9, using 35 (60 mg, 0.11 mmol) as the reactant, the target product was obtained as a yellow solid (Example 9, 10 mg, yield: 9.6%). ^¹H NMR (300 MHz, DMSO-d6 ₎ )δ11.06(s,1H),8.77(s,1H),8.05(s,1H),7.96(d,J＝4.8Hz,1H),7.56(dd,J＝5.3,3.1Hz,2H),7.38(d,J＝2.2Hz,1H),7.09(s,1H),7.03 (s,1H),6.90(dd,J＝8.6,2.1Hz,1H),5.03(dd,J＝12.7,5.3Hz,1H),4.64(s,1H),4.58(s,2H),4.48(d,J＝12.7Hz,2H),4.37(d,J＝5.4Hz, 2H),4.17(d,J＝5.4Hz,2H),3.59(s,2H),3.48(d,J＝13.8Hz,4H),3.22(s,2H),2.94(t,J＝12.8Hz,4H),2.64(d,J＝4.6Hz,3H),2.59(s,1H ),2.33(d,J＝22.3Hz,5H),1.96(dt,J＝13.4,6.6Hz,2H),1.65(d,J＝12.4Hz,2H),1.47(d,J＝10.8Hz,2H),1.23(s,1H).HRMS(ESI):calcd for C ₄₅ H ₄₈ ClN ₁₁ O ₉ [M+H] ⁺ 922.3325, found 922.3402. Purity: 95.49% by HPLC (MeOH/ _H₂O = 80:20, _tR = 4.001 min).

Synthesis Route 6:

Examples 10-11 were synthesized according to synthetic route 6.

Synthetic Route 6: Reagents and Conditions: (a) ( _PPh3 ) ₃ RhCl, EtOH, 80℃, 18h; (b) KI, pyridine, 115℃, 12h; (c) TFA, DCM, rt, 3h; (d) AcOH, NaBH(OAc) ₃ , DCE, rt, 3h; (e) TFA, DCM, rt, 3h; (f) EDCI, HOBt, DIPEA, DMF, rt, 12h.

2-(tert-butyl)5,6-dimethylisoindoline-2,5,6-tricarboxylic acid ester (37)

2-(tert-butyl)5,6-dimethyl isoindoline-2,5,6-tricarboxylate(37)

Intermediates 36a (3.40 g, 17.59 mmol) and 36b (10.00 g, 70.37 mmol) were added to a round-bottom flask containing 50 mL of anhydrous ethanol solution, followed by the addition of ( _PPh3 ) _3RhCl (163 mg, 0.18 mmol). The mixture was purged with argon and refluxed at 85 °C for 18 hours. The reaction was monitored by TLC until complete. The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (elution system: petroleum ether: ethyl acetate = 20:1, v/v) to give the target product 37 as a white solid (1.00 g, yield: 17.0%). ¹ H NMR (300MHz, DMSO-d ₆ ) δ7.72 (d, J = 9.7 Hz, 2H), 4.67 (d, J = 4.7 Hz, 4H), 3.84 (s, 6H), 1.49 (s, 9H). ESI-MS: m/z: [M+H] ⁺ 336.14.

6-(2,6-dioxopiperidin-3-yl)-5,7-dioxo-3,5,6,7-tetrahydropyrrolo[3,4-f]isoindole-2(1H)-carboxylic acid tert-butyl ester (38)

tert-butyl-6-(2,6-dioxopiperidin-3-yl)-5,7-dioxo-3,5,6,7-tetrahydropyrrolo[3,4-f]isoindole-2(1H)-carboxylate(38)

Intermediate 37 (820 mg, 2.45 mmol) and 27a (523 mg, 3.18 mmol) were added to a round-bottom flask containing 5 mL of pyridine solution, followed by the addition of potassium iodide (1.22 g, 7.34 mmol). The mixture was refluxed overnight at 115 °C, and the reaction was monitored by TLC until complete. The reaction solution was concentrated under reduced pressure and extracted with ethyl acetate and 1 M citric acid aqueous solution. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (elution system: dichloromethane:methanol = 100:1, v/v) to give the target product 38 as a yellow solid (460 mg, yield: 47.1%). ¹ H NMR (300MHz, DMSO-d ₆ )δ11.15(s,1H),7.91(d,J＝1.7Hz,2H),5.17(dd,J＝12.8,5.3Hz,1H),4.74(d,J＝7.1Hz,4H),2.92(ddd ,J＝17.0,13.8,5.4Hz,1H),2.60(t,J＝16.2Hz,2H),2.15–2.03(m,1H),1.49(s,9H).ESI-MS:m/z:[M+H] ^+400.14 .

2-(2,6-dioxopiperidin-3-yl)-6,7-dihydropyrrolo[3,4-f]isoindole-1,3(2H,5H)-dione (39a)

2-(2,6-dioxopiperidin-3-yl)-6,7-dihydropyrrolo[3,4-f]isoindole-1,3(2H,5H)-dione(39a)

Following general synthesis method 4, using 38 (400 mg, 1.00 mmol) as the reactant, the target product, a yellow oil 39a (275 mg, yield: 91.8%), was obtained. ESI-MS: m/z: [M+H] ⁺ 300.09.

4-((6-(2,6-dioxopiperidin-3-yl)-5,7-dioxo-3,5,6,7-tetrahydropyrrolo[3,4-f]isoindol-2(1H)-yl)methyl)piperidin-1-carboxylic acid tert-butyl ester (40)

tert-butyl-4-((6-(2,6-dioxopiperidin-3-yl)-5,7-dioxo-3,5,6,7-tetrahydropyrrolo[3,4-f]isoindol-2(1H)-yl)methyl)piperidine-1-carboxylate(40)

Following general synthesis method 1, using intermediate 39a (450 mg, 1.50 mmol) as the starting material, the target product, white solid 40 (345 mg, yield: 46.2%), was obtained. ¹ H NMR (300MHz, DMSO-d ₆ )δ11.13(s,1H),7.81–7.77(m,2H),5.20–5.10(m,1H),3.96(d,J＝13.6Hz,6H),3.85(d,J＝13.2Hz,4H),2.59(d,J＝ 5.4Hz,2H),2.41(t,J＝4.0Hz,1H),2.14–2.01(m,1H),1.78(t,J＝6.5Hz,5H),1.41–1.40(m,9H).ESI-MS:m/z:[M+H] ^+497.23 .

2-(2,6-dioxopiperidin-3-yl)-6-(piperidin-4-ylmethyl)-6,7-dihydropyrrolo[3,4-f]isoindole-1,3(2H,5H)-dione (41)

2-(2,6-dioxopiperidin-3-yl)-6-(piperidin-4-ylmethyl)-6,7-dihydropyrrolo[3,4-f]isoindole-1,3(2H,5H)-dione(41)

Following general synthetic method 4, using intermediate 40 (190 mg, 0.38 mmol) as the reactant, the target product, a white oil 41 (115 mg, yield: 75.8%), was obtained. ESI-MS: m/z: [M+H] ⁺ 397.18.

Example 10

2-((9-((5-chloro-2-(4-(6-(2,6-dioxopiperidin-3-yl)-5,7-dioxo-1,2,3,5,6,7-hexahydropyrrolo[3,4-f]isoindole-2-carbonyl)piperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-6-yl)oxy)-N-methylacetamide (Example 10)

2-((9-((5-chloro-2-(4-(6-(2,6-dioxopiperidin-3-yl)-5,7-dioxo-1,2,3,5,6,7-hexahydropyrrolo[3,4-f]isoindole-2-carbonyl)pip eridin-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 (Example 10)

Following general synthesis method 9, using 35 (60 mg, 0.11 mmol) as the reactant, the target product was obtained as a yellow solid (Example 10, 9 mg, yield: 9.8%). ^¹H NMR (300 MHz, DMSO- _d₆ ) δ 11.16 (s, 1H), 8.80 (s, 1H), 8.08 (s, 1H), 7.97 (d, J = 5.2 Hz, 1H), 7.94 (s, 1H), 7.89 (s, 1H), 7.61 (d, J = 2.0 Hz, 1H), 7.41 (d, J = 2.2 Hz, 1H), 7.10 (s, 1H), 5.18 (dd, J = 12.9, 4.3 Hz, 3H), 4.79 (s,2H),4.57(d,J＝11.8Hz,4H),4.39(t,J＝4.8Hz,2H),4.17(t,J＝4.8Hz,2H),3.04–2.86(m,4 H),2.63(d,J＝4.8Hz,5H),2.15–2.06(m,1H),1.84(d,J＝12.5Hz,2H),1.56(d,J＝12.3Hz,2H). ¹³ C NMR (75MHz, DMSO-d ₆ )δ173.35,173.27,170.34,167.91,167.41,159.53,155.76,155.23,155 .06,147.61,144.91,144.23,142.95,134.54,131.44,131.39,120.37,1 19.18,118.94,118.85,113.66,112.36,109.31,102.52,68.42,64.20,5 2.44,52.25,49.57,44.05,31.45,27.71,25.85,22.53.HRMS(ESI):calcd for C ₃₉ H _{36ClN 9} O ₉ [M+Na] ⁺ 832.2325, found 832.2228. Purity: 96.41% by HPLC (MeOH/H ₂ O＝80:20, t _R ＝4.301min).

Example 11

2-((9-((5-chloro-2-(4-(4-((6-(2,6-dioxopiperidin-3-yl)-5,7-dioxo-3,5,6,7-tetrahydropyrrolo[3,4-f]isoindol-2(1H)-yl)methyl)piperidin-1-carbonyl)piperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-6-yl)oxy)-N-methylacetamide (Example 11)

2-((9-((5-chloro-2-(4-(4-((6-(2,6-dioxopiperidin-3-yl)-5,7-dioxo-3,5,6,7-tetrahydropyrrolo[3,4-f]isoindol-2(1H)-yl)methyl)piperidine- 1-carbonyl)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 (Example 11)

Following general synthesis method 9, using 35 (60 mg, 0.11 mmol) as the reactant, the target product was obtained as a yellow solid (Example 11, 12 mg, yield: 11.7%). ^¹H NMR (300 MHz, DMSO- _d₆ ) δ 11.16 (s, 1H), 8.79 (s, 1H), 8.07 (d, J = 1.1 Hz, 1H), 8.00 (d, J = 5.1 Hz, 1H), 7.79 (d, J = 17.3 Hz, 2H), 7.59 (d, J = 2.2 Hz, 1H), 7.41 (d, J = 2.1 Hz, 1H), 7.12 (s, 1H), 5.16 (dd, J = 12.7, 5.3 Hz, 1H), 4.61 (s, 2H), 4.51 (d, J = 12.0 Hz). ,2H),4.40(d,J＝4.8Hz,2H),4.22–4.17(m,2H),4.00(s,4H),2.95(q,J＝15.7,14.1Hz,6H),2.70–2.66(m,4H) ,2.59(s,5H),2.13–2.05(m,1H),1.91–1.73(m,3H),1.65(s,2H),1.51(s,3H),1.26(s,1H).HRMS(ESI):calcd for C ₄₇ H ₅₂ ClN ₁₁ O ₉ [M+H] ⁺ 907.3216, found 907.3258. Purity: 97.32% by HPLC (MeOH/ _H₂O = 80:20, _tR = 4.108 min).

Synthesis Route 7:

Examples 12-16 were synthesized according to synthetic route 7.

Synthetic Route 7: Reagents and Conditions: (a) DIPEA, DMSO, 100℃, 3h; (b) TFA, DCM, rt, 1h; (c) NaBH(OAc) ₃ , DCE, rt, 5h; (d) TFA, DCM, rt, 1h; (e) DIPEA, EDCI, HOBt, DMF, rt, 12h.

6-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)-2,6-diazaspiro[3.3]heptane-2-carboxylic acid tert-butyl ester (43a)

tert-butyl-6-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-2,6-diazaspiro[3.3]heptane-2-carboxylate(43a)

Following general synthesis method 3, using 23b (400 mg, 1.45 mmol) as the reactant, the target product, a yellow solid 43a (614 mg, yield: 93.3%), was obtained. ¹ H NMR (300MHz, DMSO-d ₆ )δ10.96(s,1H),7.52(d,J＝8.3Hz,1H),6.66(d,J＝2.1Hz,1H),6.52(dd,J＝8.3,2.2Hz,1H),4.93(dd,J＝12.8,5.3Hz,1H),4.07–4.01( m,4H),3.98–3.88(m,4H),2.85–2.67(m,1H),2.48(s,1H),2.42(s,1H),1.88(d,J＝11.6Hz,1H),1.28–1.22(m,9H).ESI-MS:m/z:[M+H] ^+455.18 .

2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)-2,7-diazaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester (43b)

tert-butyl-2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-2,7-diazaspiro[3.5]nonane-7-carboxylate(43b)

Following general synthesis method 3, using 23b (400 mg, 1.45 mmol) as the reactant, the target product, a yellow solid 43b (643 mg, yield: 92.0%), was obtained. ESI-MS: m/z: [M+H] ⁺ 483.22.

7-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)-2,7-diazaspiro[4.4]nonane-2-carboxylic acid tert-butyl ester (43c)

tert-butyl-7-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-2,7-diazaspiro[4.4]nonane-2-carboxylate(43c)

Following general synthesis method 3, using 23b (400 mg, 1.45 mmol) as the reactant, the target product, a yellow solid 43c (610 mg, yield: 87.3%), was obtained. ESI-MS: m/z: [M+H] ⁺ 483.22.

2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)-2,8-diazaspiro[4.5]decane-8-carboxylic acid tert-butyl ester (43d)

tert-butyl-2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-2,8-diazaspiro[4.5]decane-8-carboxylate(43d)

Following general synthesis method 3, using 23b (400 mg, 1.45 mmol) as the reactant, the target product, a yellow solid 43d (594 mg, yield: 82.6%), was obtained. ESI-MS: m/z: [M+H] ⁺ 497.23.

9-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)-3,9-diazaspiro[5.5]undecane-3-carboxylic acid tert-butyl ester (43e)

tert-butyl-9-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-3,9-diazaspiro[5.5]undecane-3-carboxylate(43e)

Following general synthesis method 3, using 23b (400 mg, 1.45 mmol) as the reactant, the target product, a yellow solid 43e (605 mg, yield: 81.8%), was obtained. ESI-MS: m/z: [M+H] ⁺ 511.25.

2-(2,6-dioxopiperidin-3-yl)-5-(2,6-diazaspiro[3.3]heptane-2-yl)isoindoline-1,3-dione (44a)

2-(2,6-dioxopiperidin-3-yl)-5-(2,6-diazaspiro[3.3]heptan-2-yl)isoindoline-1,3-dione(44a)

Following general synthesis method 4, using 43a (300 mg, 0.59 mmol) as the reactant, the target product, a yellow oil 44a (187 mg, yield: 79.9%), was obtained. ESI-MS: m/z: [M+H] ⁺ 355.13.

2-(2,6-dioxopiperidin-3-yl)-5-(2,7-diazaspiro[3.5]nonane-2-yl)isoindoline-1,3-dione (44b)

2-(2,6-dioxopiperidin-3-yl)-5-(2,7-diazaspiro[3.5]nonan-2-yl)isoindoline-1,3-dione(44b)

Following general synthesis method 4, using 43b (300 mg, 0.62 mmol) as the reactant, the target product, a yellow oil 44b (171 mg, yield: 71.9%), was obtained. ESI-MS: m/z: [M+H] ⁺ 383.16.

2-(2,6-dioxopiperidin-3-yl)-5-(2,7-diazaspiro[4.4]nonane-2-yl)isoindoline-1,3-dione (44c)

2-(2,6-dioxopiperidin-3-yl)-5-(2,7-diazaspiro[4.4]nonan-2-yl)isoindoline-1,3-dione(44c)

Following general synthesis method 4, using 43c (300 mg, 0.62 mmol) as the reactant, the target product, a yellow oil 44c (183 mg, yield: 77.0%), was obtained. ESI-MS: m/z: [M+H] ⁺ 383.16.

2-(2,6-dioxopiperidin-3-yl)-5-(2,8-diazaspiro[4.5]decane-2-yl)isoindoline-1,3-dione (44d)

2-(2,6-dioxopiperidin-3-yl)-5-(2,8-diazaspiro[4.5]decan-2-yl)isoindoline-1,3-dione(44d)

Following general synthesis method 4, using 43d (300 mg, 0.60 mmol) as the reactant, the target product, a yellow oil 44d (212 mg, yield: 88.5%), was obtained. ESI-MS: m/z: [M+H] ⁺ 397.18.

2-(2,6-dioxopiperidin-3-yl)-5-(3,9-diazaspiro[5.5]undecane-3-yl)isoindoline-1,3-dione (44e)

2-(2,6-dioxopiperidin-3-yl)-5-(3,9-diazaspiro[5.5]undecan-3-yl)isoindoline-1,3-dione(44e)

Following general synthesis method 4, using 43e (300 mg, 0.59 mmol) as the reactant, the target product, a yellow oily substance 44e (215 mg, yield: 89.2%), was obtained. ESI-MS: m/z: [M+H] ⁺ 411.20.

4-((6-(2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)-2,6-diazaspiro[3.3]heptane-2-yl)methyl)piperidin-1-carboxylic acid tert-butyl ester (45a)

tert-butyl-4-((6-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-2,6-diazaspiro[3.3]heptan-2-yl)methyl)piperidine-1-carboxylate(45a)

Following general synthetic method 1, using intermediate 44a (300 mg, 0.85 mmol) as the reactant, the target product, a yellow solid 45a (248 mg, yield: 53.1%), was obtained. ESI-MS: m/z: [M+H] ⁺ 552.27.

4-((2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)-2,7-diazaspiro[3.5]nonane-7-yl)methyl)piperidin-1-carboxylic acid tert-butyl ester (45b)

tert-butyl-4-((2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-2,7-diazaspiro[3.5]nonan-7-yl)methyl)piperidine-1-carboxylate(45b)

Following general synthetic method 1, using intermediate 44b (300 mg, 0.78 mmol) as the reactant, the target product, a yellow solid 45b (263 mg, yield: 57.8%), was obtained. ESI-MS: m/z: [M+H] ⁺ 580.30.

4-((7-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)-2,7-diazaspiro[4.4]nonane-2-yl)methyl)piperidin-1-carboxylic acid tert-butyl ester (45c)

tert-butyl-4-((7-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-2,7-diazaspiro[4.4]nonan-2-yl)methyl)piperidine-1-carboxylate(45c)

Following general synthetic method 1, using intermediate 44c (300 mg, 0.78 mmol) as the reactant, the target product, a yellow solid 45c (211 mg, yield: 46.4%), was obtained. ESI-MS: m/z: [M+H] ⁺ 580.30.

4-((2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)-2,8-diazaspiro[4.5]decane-8-yl)methyl)piperidin-1-carboxylic acid tert-butyl ester (45d)

tert-butyl-4-((2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-2,8-diazaspiro[4.5]decan-8-yl)methyl)piperidine-1-carboxylate(45d)

Following general synthesis method 1, using intermediate 44d (300 mg, 0.76 mmol) as the reactant, the target product, a yellow solid 45d (224 mg, yield: 49.9%), was obtained. ^¹H NMR (300 MHz, DMSO- _d⁶ ) δ 11.20 (s, 1H), 7.75 (d, J = 8.4 Hz, 1H), 7.01 (d, J = 2.1 Hz, 1H), 6.91 (dd, J = 8.6, 2.2 Hz, 1H), 5.17 (dd, J = 12.5, 5.3 Hz, 1H), 3.97 (s, 2H), 3.57 (s, 3H), 3.31–3.25 (m, 1H), 2.98 (d, 2.5 mmol, 0.76 ... J＝12.4Hz,1H),2.71(s,2H),2.66(s,2H),2.35(s,2H),2.19(d,J＝6.7Hz,5H),1.95–1.8 7(m,1H),1.71(d,J＝16.3Hz,7H),1.50(d,J＝2.0Hz,3H),1.48(s,9H).ESI-MS:m/z:[M+H] ^+594.32 .

4-((9-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)-3,9-diazaspiro[5.5]undecane-3-yl)methyl)piperidin-1-carboxylic acid tert-butyl ester (45e)

tert-butyl-4-((9-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)-3,9-diazaspiro[5.5]undecan-3-yl)methyl)piperidine-1-carboxylate(45e)

Following general synthetic method 1, using intermediate 44e (300 mg, 0.73 mmol) as the reactant, the target product, a yellow solid 45e (232 mg, yield: 52.2%), was obtained. ESI-MS: m/z: [M+H] ⁺ 608.34.

2-(2,6-dioxopiperidin-3-yl)-5-(6-(piperidin-4-ylmethyl)-2,6-diazaspiro[3.3]heptane-2-yl)isoindoline-1,3-dione (46a)

2-(2,6-dioxopiperidin-3-yl)-5-(6-(piperidin-4-ylmethyl)-2,6-diazaspiro[3.3]heptan-2-yl)isoindoline-1,3-dione(46a)

Following general synthesis method 4, using 45a (300 mg, 0.54 mmol) as the reactant, the target product, a yellow oil 46a (213 mg, yield: 86.7%), was obtained. ESI-MS: m/z: [M+H] ⁺ 452.22.

2-(2,6-dioxopiperidin-3-yl)-5-(7-(piperidin-4-ylmethyl)-2,7-diazaspiro[3.5]nonane-2-yl)isoindoline-1,3-dione (46b)

2-(2,6-dioxopiperidin-3-yl)-5-(7-(piperidin-4-ylmethyl)-2,7-diazaspiro[3.5]nonan-2-yl)isoindoline-1,3-dione(46b)

Following general synthesis method 4, using 45b (300 mg, 0.52 mmol) as the reactant, the target product, a yellow oil 46b (233 mg, yield: 93.9%), was obtained. ESI-MS: m/z: [M+H] ⁺ 480.25.

2-(2,6-dioxopiperidin-3-yl)-5-(7-(piperidin-4-ylmethyl)-2,7-diazaspiro[4.4]nonane-2-yl)isoindoline-1,3-dione (46c)

2-(2,6-dioxopiperidin-3-yl)-5-(7-(piperidin-4-ylmethyl)-2,7-diazaspiro[4.4]nonan-2-yl)isoindoline-1,3-dione(46c)

Following general synthesis method 4, using 45c (300 mg, 0.54 mmol) as the reactant, the target product, a yellow oil 46c (221 mg, yield: 89.0%), was obtained. ESI-MS: m/z: [M+H] ⁺ 480.25.

2-(2,6-dioxopiperidin-3-yl)-5-(8-(piperidin-4-ylmethyl)-2,8-diazaspiro[4.5]decane-2-yl)isoindoline-1,3-dione (46d)

2-(2,6-dioxopiperidin-3-yl)-5-(8-(piperidin-4-ylmethyl)-2,8-diazaspiro[4.5]decan-2-yl)isoindoline-1,3-dione(46d)

Following general synthesis method 4, using 45d (300 mg, 0.50 mmol) as the reactant, the target product, a yellow oily substance 46d (217 mg, yield: 87.0%), was obtained. ESI-MS: m/z: [M+H] ⁺ 494.27.

2-(2,6-dioxopiperidin-3-yl)-5-(9-(piperidin-4-ylmethyl)-3,9-diazaspiro[5.5]undecane-3-yl)isoindoline-1,3-dione (46e)

2-(2,6-dioxopiperidin-3-yl)-5-(9-(piperidin-4-ylmethyl)-3,9-diazaspiro[5.5]undecan-3-yl)isoindoline-1,3-dione(46e)

Following general synthetic method 4, using intermediate 45e (300 mg, 0.73 mmol) as the reactant, the target product, a yellow solid 46e (232 mg, yield: 52.2%), was obtained. ESI-MS: m/z: [M+H] ⁺ 508.28.

Example 12

2-((9-((5-chloro-2-(4-(4-((6-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)-2,6-diazaspiro[3.3]heptane-2-yl)methyl)piperidin-1-carbonyl)piperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-6-yl)oxy)-N-methylacetamide (Example 12)

2-((9-((5-chloro-2-(4-(4-((6-(2-(2,6-dioxopiperidin-3-yl))-1,3-dioxoisoindolin-5-yl)-2,6-diazaspiro[3.3]heptan-2-yl)methyl)piperidine- 1-carbonyl)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 (Example 12)

Following general synthesis method 9, using 46a (60 mg, 0.11 mmol) as the reactant, the target product was obtained as a yellow solid (Example 12, 13 mg, yield: 11.9%). ^¹H NMR (300 MHz, DMSO- _d6) was performed. )δ11.10(s,1H),8.78(s,1H),8.07(s,1H),7.98(q,J＝4.6,3.8Hz,1H),7.66(d,J＝8.3Hz,1H),7.58(d,J＝2.2Hz,1H),7.40(d,J＝2.2Hz,1H ),7.12(s,1H),6.80(d,J＝2.0Hz,1H),6.66(dd,J＝8.4,2.0Hz,1H),5.07(dd,J＝12.8,5.4Hz,1H),4.60(s,2H),4.50(d,J＝12.4Hz,2H),4. 39(d,J＝5.3Hz,2H),4.34(s,1H),4.22–4.16(m,2H),4.12(s,4H),4.01(d,J＝12.7Hz,1H),3.30(s,5H),3.08–2.88(m,5H),2.67(d,J＝4.6 Hz,3H),2.62(s,1H),2.56(s,1H),2.28(s,2H),2.08–1.96(m,1H),1.80–1.59(m,4H),1.59–1.36(m,3H),1.26(s,2H).HRMS(ESI):calcd for C ₄₈ H ₅₂ ClN ₁₁ O ₉ [M+H] ⁺ 962.3638, found 962.3704. Purity: 95.22% by HPLC (MeOH/H ₂ O = 80:20, t _R = 3.901 min).

Example 13

2-((9-((5-chloro-2-(4-(4-((2-(2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)-2,7-diazaspiro[3.5]nonane-7-yl)methyl)piperidin-1-carbonyl)piperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-6-yl)oxy)-N-methylacetamide (Example 13)

2-((9-((5-chloro-2-(4-(4-((2-(2-(2,6-dioxopiperidin-3-yl))-1,3-dioxoisoindolin-5-yl)-2,7-diazaspiro[3.5]nonan-7-yl)methyl)piperidine-1 -carbonyl)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 (Example 13)

Following general synthetic method 9, using 46b (60 mg, 0.11 mmol) as the reactant, the target product was obtained as a yellow solid (Example 13, 8 mg, yield: 7.1%). ^¹H NMR (300 MHz, DMSO-d6 ₎ )δ11.08(s,1H),8.77(s,1H),8.05(s,1H),7.97(d,J＝4.8Hz,1H),7.63(d,J＝8.2Hz,1H),7.56(d,J＝2.2Hz,1H),7.38(d,J＝2.2Hz,1H),7 .10(s,1H),6.77(d,J＝2.0Hz,1H),6.64(dd,J＝8.4,2.1Hz,1H),5.05(dd,J＝12.7,5.4Hz,1H),4.58(s,2H),4.48(d,J＝11.7Hz,2H),4.38( s,2H),4.17(d,J＝4.8Hz,2H),4.01(d,J＝12.8Hz,1H),3.74(s,4H),3.05–2.80(m,6H),2.65(d,J＝4.6Hz,3H),2.60(s,1H),2.54(s,3H),2 .29(d,J＝11.7Hz,3H),2.11(s,2H),2.02–1.96(m,1H),1.75(s,6H),1.63(d,J＝14.0Hz,3H),1.47(s,2H),1.23(s,1H).HRMS(ESI):calcd for C ₅₀ H ₅₆ ClN ₁₁ O ₉ [M+H] ⁺ 990.3951,found 990.4034. Purity: 96.64% by HPLC (MeOH/H ₂ O = 80:20, t _R = 3.817 min).

Example 14

2-((9-((5-chloro-2-(4-(4-((7-(2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)-2,7-diazaspiro[4.4]nonane-2-yl)methyl)piperidin-1-carbonyl)piperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-6-yl)oxy)-N-methylacetamide (Example 14)

2-((9-((5-chloro-2-(4-(4-((7-(2-(2,6-dioxopiperidin-3-yl))-1,3-dioxoisoindolin-5-yl)-2,7-diazaspiro[4.4]nonan-2-yl)methyl)piperidine-1 -carbonyl)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 (Example 14)

Following general synthetic method 9, using 46c (60 mg, 0.11 mmol) as the reactant, the target product was obtained as a yellow solid (Example 14, 14 mg, yield: 12.5%). ^¹H NMR (300 MHz, DMSO- _d6) was performed. )δ10.87(s,1H),8.56(s,1H),7.83(s,1H),7.76(d,J＝4.9Hz,1H),7.43(d,J＝8.4Hz,1H),7.35(d,J＝2.2Hz,1H),7.17(d,J＝2.2Hz, 1H),6.88(s,1H),6.68(s,1H),6.58(d,J＝8.7Hz,1H),4.84(dd,J＝12.5,5.4Hz,1H),4.36(s,2H),4.27(d,J＝12.2Hz,2H),4.19–4. 15(m,2H),4.12(s,1H),3.98–3.92(m,2H),3.79(d,J＝12.4Hz,1H),3.23(d,J＝6.7Hz,3H),3.17(s,2H),2.85–2.60(m,6H),2.43(d ,J＝4.6Hz,5H),2.33(s,5H),2.06(s,1H),1.86–1.74(m,3H),1.62–1.38(m,7H),1.31–1.20(m,2H),1.02(s,1H).HRMS(ESI):calcd for C ₅₀ H ₅₆ ClN ₁₁ O ₉ [M+H] ⁺ 990.3951,found 990.4016. Purity: 95.82% by HPLC (MeOH/ _H₂O = 80:20, _tR = 3.842 min).

Example 15

2-((9-((5-chloro-2-(4-(4-((2-(2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)-2,8-diazaspiro[4.5]decane-8-yl)methyl)piperidin-1-carbonyl)piperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-6-yl)oxy)-N-methylacetamide (Example 15)

2-((9-((5-chloro-2-(4-(4-((2-(2-(2,6-dioxopiperidin-3-yl))-1,3-dioxoisoindolin-5-yl)-2,8-diazaspiro[4.5]decan-8-yl)methyl)piperidine-1 -carbonyl)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 (Example 15)

Following general synthetic method 9, using 46d (60 mg, 0.11 mmol) as the reactant, the target product was obtained as a yellow solid (Example 15, 11 mg, yield: 9.6%). ^¹H NMR (300 MHz, DMSO- _d6) was performed. )δ11.10(s,1H),8.79(s,1H),8.10–8.05(m,1H),8.00(d,J＝4.8Hz,1H),7.66 (d,J＝8.5Hz,1H),7.59(d,J＝2.2Hz,1H),7.40(d,J＝2.2Hz,1H),7.12(s,1H),6 .94(d,J＝2.1Hz,1H),6.83(dd,J＝8.7,2.1Hz,1H),5.08(dd,J＝12.6,5.4Hz,1H ),4.60(s,2H),4.51(d,J＝12.5Hz,2H),4.41(d,J＝4.5Hz,2H),4.37(s,1H),4. 19(t,J＝4.7Hz,2H),4.03(d,J＝12.7Hz,1H),3.49(s,3H),3.40(s,3H),3.31(s ,1H),3.10–2.85(m,6H),2.67(d,J＝4.6Hz,3H),2.62(s,1H),2.57(s,3H),2.4 0–2.12(m,2H),2.06–1.98(m,1H),1.94–1.87(m,2H),1.79(d,J＝13.8Hz,3H), 1.65(d,J＝12.0Hz,6H),1.51(d,J＝6.6Hz,2H),1.26(s,1H).HRMS(ESI):calcd for C ₅₁ H ₅₈ ClN ₁₁ O ₉ [M+H] ⁺ 1004.4108, found 1004.4176. Purity: 95.02% by HPLC (MeOH/H ₂ O = 80:20, t _R = 3.813 min).

Example 16

2-((9-((5-chloro-2-(4-(4-((9-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)-3,9-diazaspiro[5.5]undecane-3-yl)methyl)piperidin-1-carbonyl)piperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-6-yl)oxy)-N-methylacetamide (Example 16)

2-((9-((5-chloro-2-(4-(4-((9-(2-(2,6-dioxopiperidin-3-yl))-1,3-dioxoisoindolin-5-yl)-3,9-diazaspiro[5.5]undecan-3-yl)methyl)piperidine- 1-carbonyl)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 (Example 16)

Following general synthetic method 9, using 46e (60 mg, 0.11 mmol) as the reactant, the target product was obtained as a yellow solid (Example 16, 17 mg, yield: 14.7%). ^¹H NMR (300 MHz, DMSO- _d6) was used. )δ11.12(s,1H),8.80(s,1H),8.07(s,1H),8.00(q,J＝4.2Hz,1H),7.67(d,J＝8.5Hz,1H),7.59(d,J＝2.2Hz,1H),7.40(d,J＝2.0Hz,1H),7 .32(d,J＝2.1Hz,1H),7.24(d,J＝8.9Hz,1H),7.12(s,1H),5.09(dd,J＝12.8,5.4Hz,1H),4.60(s,2H),4.51(d,J＝12.4Hz,2H),4.40(t,J＝4 .3Hz,2H),4.36(s,1H),4.18(t,J＝4.2Hz,2H),4.02(d,J＝12.3Hz,1H),3.47(s,6H),2.96(s,2H),2.67(d,J＝4.6Hz,3H),2.63(s,1H),2. 57(s,2H),2.36(s,4H),2.17(s,2H),2.06–1.98(m,2H),1.77(d,J＝14.7Hz,4H),1.65(d,J＝13.8Hz,2H),1.52(s,11H).HRMS(ESI):calcd for C ₅₂ H ₆₀ ClN ₁₁ O ₉ [M+H] ⁺ 1018.4264, found 1018.4340. Purity: 95.29% by HPLC (MeOH/H ₂ O = 80:20, t _R = 3.702 min).

Synthesis Route 8:

Examples 17-18 were synthesized according to synthetic route 8.

Synthetic Route 8: Reagents and Conditions: (a _{) Cs₂CO₃} , Xantphos, _Pd₂ (dba) _₃ , dioxane, 100℃ for 4h; ( _{b) Cs₂CO₃, PdCl₂(dppf)CH₂Cl₂, dioxane, 100℃ for 4h; (c) H₂ , Pd / C ,} THF, 12h; (d) TFA, DCM, 3h; (e) NaBH(OAc) _₃ , DCE, 6h; (f) TFA, DCM, 3h; (g) EDCI, HOBt, DIPEA, DMF, 12h.

7-(4-bromo-2-fluorophenyl)-2,7-diazaspiro[4.4]nonane-2-carboxylic acid tert-butyl ester (48a)

tert-butyl 7-(4-bromo-2-fluorophenyl)-2,7-diazaspiro[4.4]nonane-2-carboxylate(48a)

Synthetic Method 10: Intermediate 47 (500 mg, 1.66 mmol) and 47a (376 mg, 1.66 mmol) were added to a sealed tube containing 10 mL of anhydrous dioxane solution. Cesium carbonate (1.08 g, 3.32 mmol), _Pd₂ (dba) _₃ (106 mg, 0.12 mmol), and XantPhos (144 mg, 0.25 mmol) were then added sequentially. The mixture was purged with argon and reacted at 100 °C for 4 hours. The reaction was monitored by TLC until complete. After cooling to room temperature, the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (elution system: dichloromethane:methanol = 100:1, v/v) to obtain intermediate 48a as a white solid (560 mg, yield: 84.4%). ESI-MS: m/z: [M+H] ⁺ 399.10.

2-(4-bromo-2-fluorophenyl)-2,8-diazaspiro[4.5]decane-8-carboxylic acid tert-butyl ester (48b)

tert-butyl 2-(4-bromo-2-fluorophenyl)-2,8-diazaspiro[4.5]decane-8-carboxylate(48b)

Following general synthetic method 10, using 47b (500 mg, 1.66 mmol) as the reactant, the target product, white solid 48b (490 mg, yield: 71.3%), was obtained. ESI-MS: m/z: [M+H] ⁺ 413.12.

7-(4-(2,6-bis(benzyloxy)pyridin-3-yl)-2-fluorophenyl)-2,7-diazaspiro[4.4]nonane-2-carboxylic acid tert-butyl ester (50a)

tert-butyl-7-(4-(2,6-bis(benzyloxy)pyridin-3-yl)-2-fluorophenyl)-2,7-diazaspiro[4.4]nonane-2-carboxylate(50a)

Synthetic Method 11: Intermediates 48a (560 mg, 1.40 mmol) and 49 (761 mg, 1.82 mmol) were added to a sealed tube containing a 6 mL:2 mL dioxane:water mixture. Cesium carbonate (1.08 g, 3.32 mmol) and _PdCl₂ (dppf) _CH₂Cl₂ (110 mg, 0.14 mmol) were then added sequentially. The _mixture was purged with argon and reacted at 100 °C for 4 hours. The reaction was monitored by TLC until complete. After cooling to room temperature, the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (elution system: petroleum ether:ethyl acetate = 5:1, v/v) to give intermediate 50a as a white solid (750 mg, yield: 87.7%). ^¹H NMR (300 MHz, DMSO- _d₆) was performed. )δ7.77(d,J＝8.2Hz,1H),7.50(d,J＝2.0Hz,1H),7.47(q,J＝1.9Hz,2H),7.45(t,J＝1.9Hz,2H),7.42–7.40(m ,2H),7.38(d,J＝1.7Hz,2H),7.37(d,J＝1.3Hz,1H),7.35–7.32(m,1H),7.28(dd,J＝8.4,2.1Hz,1H),6.76(dd ,J＝9.9,8.5Hz,1H),6.57(d,J＝8.1Hz,1H),5.43(d,J＝12.6Hz,4H),3.49(td,J＝7.0,2.6Hz,2H),3.41(s,1H) ,3.35(d,J＝6.3Hz,3H),3.28(d,J＝4.4Hz,2H),1.98–1.85(m,4H),1.45(d,J＝2.7Hz,9H).ESI-MS:m/z:[M+H] ^+610.30 .

2-(4-(2,6-bis(benzyloxy)pyridin-3-yl)-2-fluorophenyl)-2,8-diazaspiro[4.5]decane-8-carboxylic acid tert-butyl ester (50b)

tert-butyl-2-(4-(2,6-bis(benzyloxy)pyridin-3-yl)-2-fluorophenyl)-2,8-diazaspiro[4.5]decane-8-carboxylate(50b)

Following general synthetic method 11, using 48b (540 mg, 1.31 mmol) as the reactant, the target product, a white solid 50b (710 mg, yield: 87.1%), was obtained. ESI-MS: m/z: [M+H] ⁺ 624.32.

7-(4-(2,6-dioxopiperidin-3-yl)-2-fluorophenyl)-2,7-diazaspiro[4.4]nonane-2-carboxylic acid tert-butyl ester (51a)

tert-butyl-7-(4-(2,6-dioxopiperidin-3-yl)-2-fluorophenyl)-2,7-diazaspiro[4.4]nonane-2-carboxylate(51a)

Synthetic Method 12: Intermediate 50a (750 mg, 1.23 mmol) was added to a round-bottom flask containing 10 mL of tetrahydrofuran solution. Palladium on carbon (150 mg, 20% m/m) was added, and the mixture was purged with hydrogen. The mixture was stirred at room temperature for 16 hours, and the reaction was monitored by TLC until complete. The reaction solution was filtered through diatomaceous earth, washed with dichloromethane, and the filtrate was concentrated under low pressure. The residue was purified by column chromatography (elution system: dichloromethane:methanol = 100:1, v/v) to give intermediate 51a as a white oil (451 mg, yield: 85.1%). ESI-MS: m/z: [M+H] ⁺ 432.22.

2-(4-(2,6-dioxopiperidin-3-yl)-2-fluorophenyl)-2,8-diazaspiro[4.5]decane-8-carboxylic acid tert-butyl ester (51b)

tert-butyl-2-(4-(2,6-dioxopiperidin-3-yl)-2-fluorophenyl)-2,8-diazaspiro[4.5]decane-8-carboxylate(51b)

Following general synthetic method 12, using 50b (680 mg, 1.09 mmol) as the reactant, the target product, a white oily substance 51b (462 mg, yield: 95.1%), was obtained. ESI-MS: m/z: [M+H] ⁺ 446.24.

3-(3-fluoro-4-(2,7-diazaspiro[4.4]nonane-2-yl)phenyl)piperidine-2,6-dione (52a)

3-(3-fluoro-4-(2,7-diazaspiro[4.4]nonan-2-yl)phenyl)piperidine-2,6-dione(52a)

Following general synthesis method 4, using 51a (440 mg, 1.02 mmol) as the reactant, the target product, a white oily substance 52a (315 mg, yield: 93.2%), was obtained. ESI-MS: m/z: [M+H] ⁺ 332.17.

3-(3-fluoro-4-(2,8-diazaspiro[4.5]decane-2-yl)phenyl)piperidin-2,6-dione (52b)

3-(3-fluoro-4-(2,8-diazaspiro[4.5]decan-2-yl)phenyl)piperidine-2,6-dione(52b)

Following general synthesis method 4, using 51b (430 mg, 0.96 mmol) as the reactant, the target product, a white oily substance 52b (322 mg, yield: 96.6%), was obtained. ESI-MS: m/z: [M+H] ⁺ 346.18.

4-((7-(4-(2,6-dioxopiperidin-3-yl)-2-fluorophenyl)-2,7-diazaspiro[4.4]nonane-2-yl)methyl)piperidin-1-carboxylic acid tert-butyl ester (53a)

tert-butyl-4-((7-(4-(2,6-dioxopiperidin-3-yl)-2-fluorophenyl)-2,7-diazaspiro[4.4]nonan-2-yl)methyl)piperidine-1-carboxylate(53a)

Following general synthetic method 1, using intermediate 52a (300 mg, 0.90 mmol) as the reactant, the target product, white solid 53a (302 mg, yield: 63.1%), was obtained. ESI-MS: m/z: [M+H] ⁺ 529.31.

4-((2-(4-(2,6-dioxopiperidin-3-yl)-2-fluorophenyl)-2,8-diazaspiro[4.5]decane-8-yl)methyl)piperidin-1-carboxylic acid tert-butyl ester (53b)

tert-butyl-4-((2-(4-(2,6-dioxopiperidin-3-yl)-2-fluorophenyl)-2,8-diazaspiro[4.5]decan-8-yl)methyl)piperidine-1-carboxylate(53b)

Following general synthetic method 1, using intermediate 52b (300 mg, 0.87 mmol) as the reactant, the target product, white solid 53b (290 mg, yield: 61.5%), was obtained. ESI-MS: m/z: [M+H] ⁺ 543.33.

3-(3-fluoro-4-(7-(piperidin-4-ylmethyl)-2,7-diazaspiro[4.4]nonane-2-yl)phenyl)piperidin-2,6-dione (54a)

3-(3-fluoro-4-(7-(piperidin-4-ylmethyl)-2,7-diazaspiro[4.4]nonan-2-yl)phenyl)piperidine-2,6-dione(54a)

Following general synthesis method 4, using 53a (290 mg, 0.55 mmol) as the reactant, the target product, a white oily substance 54a (210 mg, yield: 89.3%), was obtained. ESI-MS: m/z: [M+H] ⁺ 429.26.

3-(3-fluoro-4-(8-(piperidin-4-ylmethyl)-2,8-diazaspiro[4.5]decane-2-yl)phenyl)piperidin-2,6-dione (54b)

3-(3-fluoro-4-(8-(piperidin-4-ylmethyl)-2,8-diazaspiro[4.5]decan-2-yl)phenyl)piperidine-2,6-dione(54b)

Following general synthesis method 4, using 53b (270 mg, 0.50 mmol) as the reactant, the target product, a white oily substance 54b (192 mg, yield: 87.2%), was obtained. ESI-MS: m/z: [M+H] ⁺ 443.27.

Example 17

2-((9-((5-chloro-2-(4-(4-(4-((7-(4-(2,6-dioxopiperidin-3-yl)-2-fluorophenyl)-2,7-diazaspiro[4.4]nonane-2-yl)methyl)piperidin-1-carbonyl)piperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-6-yl)oxy)-N-methylacetamide (Example 17)

2-((9-((5-chloro-2-(4-(4-((7-(4-(2,6-dioxopiperidin-3-yl))-2-fluorophenyl)-2,7-diazaspiro[4.4]nonan-2-yl)methyl)piperidine-1-carb onyl)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 (Example 17)

Following general synthetic method 9, using 54a (60 mg, 0.11 mmol) as the reactant, the target product, a white solid 17 (10 mg, yield: 9.4%), was obtained. ^¹H NMR (300 MHz, DMSO- _d6) )δ10.80(s,1H),8.79(s,1H),8.06(s,1H),7.99(d,J＝5.1Hz,1H),7.58( d,J＝2.2Hz,1H),7.40(d,J＝2.2Hz,1H),7.12(s,1H),7.02–6.83(m,2H),6 .65(t,J＝9.0Hz,1H),4.60(s,2H),4.50(d,J＝12.6Hz,2H),4.40(t,J＝4.6 Hz,2H),4.35(s,1H),4.18(t,J＝4.5Hz,2H),4.03(d,J＝13.0Hz,1H),3.74 (dd,J＝11.4,4.8Hz,1H),3.39(s,3H),3.33(s,4H),3.25(d,J＝8.3Hz,1H),3.00(dt,J＝23.8,13.0Hz,5H),2.67(d,J＝4.7Hz,4H),2.61(d,J＝5.5Hz ,3H),2.34–2.11(m,2H),2.03–1.89(m,3H),1.85–1.73(m,4H),1.64(d,J＝9.6Hz,2H),1.56–1.43(m,2H),1.26(d,J＝3.6Hz,2H).HRMS(ESI):calcd for C ₄₈ H ₅₆ ClFN ₁₀ O ₇ [M+H] ⁺ 939.4006, found 939.4068. Purity: 95.55% by HPLC (MeOH/H ₂ O = 80:20, t _R = 3.782 min).

Example 18

2-((9-((5-chloro-2-(4-(4-((2-(4-(2-(2-(2,6-dioxopiperidin-3-yl)-2-fluorophenyl)-2,8-diazaspiro[4.5]decane-8-yl)methyl)piperidin-1-carbonyl)piperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-6-yl)oxy)-N-methylacetamide (Example 18)

2-((9-((5-chloro-2-(4-(4-((2-(4-(2,6-dioxopiperidin-3-yl))-2-fluorophenyl)-2,8-diazaspiro[4.5]decan-8-yl)methyl)piperidine-1-carb onyl)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 (Example 18)

Following general synthetic method 9, using 54b (60 mg, 0.11 mmol) as the reactant, the target product was obtained as a white solid (Example 18, 7 mg, yield: 6.5%). ^¹H NMR (300 MHz, DMSO- _d6) was used. )δ10.81(s,1H),8.79(s,1H),8.06(s,1H),7.99(d,J＝5.3Hz,1H),7.58(d,J ＝2.2Hz,1H),7.40(d,J＝2.2Hz,1H),7.12(s,1H),6.94(dd,J＝15.0,2.0Hz,1 H),6.86(d,J＝8.5Hz,1H),6.66(t,J＝9.1Hz,1H),4.60(s,2H),4.50(d,J＝12 .2Hz,2H),4.42–4.38(m,2H),4.36(s,1H),4.21–4.16(m,2H),4.02(d,J＝13 .0Hz,1H),3.74(dd,J＝11.4,4.8Hz,1H),3.46(s,4H),3.16(s,2H),2.98(q, J＝11.7Hz,4H),2.67(d,J＝4.6Hz,4H),2.60(d,J＝4.7Hz,1H),2.35(d,J＝20. 4Hz,3H),2.17(d,J＝12.2Hz,3H),2.04–1.97(m,1H),1.82–1.72(m,5H),1.6 5(d,J＝14.0Hz,3H),1.53(d,J＝19.7Hz,6H),1.25(s,1H).HRMS(ESI):calcd for C ₄₉ H ₅₈ ClFN ₁₀ O ₇ [M+H] ⁺ 953.4163, found 953.4231.

Synthesis Route 9:

Example 19 was synthesized according to synthetic route 9.

Synthetic Route 9: Reagents and Conditions: (a) _K₂CO₃ , DMF, 60℃, 4h; (b _{) H₂} , Pd/C, EtOH, rt, 6h; (c) _NaHCO₃ , DMF, 80℃, 16h; (d) TFA, DCM, rt, 3h; (e) NaBH(OAc) _₃ , DCE, rt, 5h; (f) TFA, DCM, rt, 2h; (g) DIPEA, EDCI, HOBt, rt, 12h.

2-(2-fluoro-4-nitrophenyl)-2,8-diazaspiro[4.5]decane-8-carboxylic acid tert-butyl ester (56)

tert-butyl 2-(2-fluoro-4-nitrophenyl)-2,8-diazaspiro[4.5]decane-8-carboxylate(56)

Synthetic Method 13: 55 (400 mg, 2.51 mmol), 47b (604 mg, 2.51 mmol), and potassium carbonate (695 mg, 5.03 mmol) were added to a round-bottom flask, followed by 10 mL of DMF solution. The mixture was stirred at 60 °C for 4 h, and the reaction was monitored by TLC until complete. After cooling to room temperature, the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (elution system: petroleum ether: ethyl acetate = 4:1, v/v) to give intermediate 56 as a white solid (874 mg, yield: 91.6%). ESI-MS: m/z: [M+H] ⁺ 380.19.

2-(4-amino-2-fluorophenyl)-2,8-diazaspiro[4.5]decane-8-carboxylic acid tert-butyl ester (57)

tert-butyl 2-(4-amino-2-fluorophenyl)-2,8-diazaspiro[4.5]decane-8-carboxylate(57)

Following general synthesis method 2, using intermediate 56 (800 mg, 2.11 mmol) as the reactant, the target product, a white oily substance 57 (719 mg, yield: 97.6%), was obtained. ESI-MS: m/z: [M+H] ⁺ 350.22.

2-(4-((2,6-dioxopiperidin-3-yl)amino)-2-fluorophenyl)-2,8-diazaspiro[4.5]decane-8-carboxylic acid tert-butyl ester (58)

tert-butyl-2-(4-((2,6-dioxopiperidin-3-yl)amino)-2-fluorophenyl)-2,8-diazaspiro[4.5]decane-8-carboxylate(58)

Synthetic Method 14: 57 (800 mg, 2.29 mmol), 57a (1.10 g, 5.72 mmol), and sodium bicarbonate (1.92 g, 22.90 mmol) were added to a round-bottom flask, followed by 15 mL of DMF solution. The mixture was stirred at 85 °C for 16 h, and the reaction was monitored by TLC until complete. After cooling to room temperature, the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (elution system: dichloromethane:methanol = 50:1, v/v) to give intermediate 58 as a white solid (632 mg, yield: 59.9%). ESI-MS: m/z: [M+H] ⁺ 461.25.

3-((3-fluoro-4-(2,8-diazaspiro[4.5]decane-2-yl)phenyl)amino)piperidine-2,6-dione (59)

3-((3-fluoro-4-(2,8-diazaspiro[4.5]decan-2-yl)phenyl)amino)piperidine-2,6-dione(59)

Following general synthesis method 4, using 58 (600 mg, 1.30 mmol) as the reactant, the target product, a white oily substance 59 (431 mg, yield: 91.8%), was obtained. ESI-MS: m/z: [M+H] ⁺ 361.20.

4-((2-(4-((2,6-dioxopiperidin-3-yl)amino)-2-fluorophenyl)-2,8-diazaspiro[4.5]decane-8-yl)methyl)piperidin-1-carboxylic acid tert-butyl ester (60)

tert-butyl-4-((2-(4-((2,6-dioxopiperidin-3-yl)amino)-2-fluorophenyl)-2,8-diazaspiro[4.5]decan-8-yl)methyl)piperidine-1-carboxylate(60)

Following general synthetic method 1, using intermediate 59 (400 mg, 1.11 mmol) as the reactant, the target product, white solid 60 (301 mg, yield: 48.6%), was obtained. ESI-MS: m/z: [M+H] ⁺ 558.34.

3-((3-fluoro-4-(8-(piperidin-4-ylmethyl)-2,8-diazaspiro[4.5]decane-2-yl)phenyl)amino)piperidin-2,6-dione (61)

3-((3-fluoro-4-(8-(piperidin-4-ylmethyl)-2,8-diazaspiro[4.5]decan-2-yl)phenyl)amino)piperidine-2,6-dione(61)

Following general synthesis method 4, using 60 (300 mg, 0.54 mmol) as the reactant, the target product, a white oily substance 61 (208 mg, yield: 84.5%), was obtained. ESI-MS: m/z: [M+H] ⁺ 458.28.

Example 19

2-((9-((5-chloro-2-(4-(4-((2-(4-((4-((2,6-dioxopiperidin-3-yl)amino)-2-fluorophenyl)-2,8-diazaspiro[4.5]decane-8-yl)methyl)piperidin-1-carbonyl)piperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-6-yl)oxy)-N-methylacetamide (Example 19)

2-((9-((5-chloro-2-(4-(4-((2-(4-((2,6-dioxopiperidin-3-yl)amino))-2-fluorophenyl)-2,8-diazaspiro[4.5]decan-8-yl)methyl)piperidine-1- carbonyl)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 (Example 19)

Following general synthetic method 9, using 61 (60 mg, 0.11 mmol) as the reactant, the target product, a brown solid 19 (9 mg, yield: 8.2%), was obtained. ^¹H NMR (300 MHz, DMSO- _d6) )δ10.79(s,1H),8.79(s,1H),8.07(s,1H),7.99(d,J＝4.4Hz,1H),7.58(d,J＝2.3Hz,1H),7.40(d,J＝2.3Hz,1H),7.12(s,1H),6.66–6.48(m,2H) ,6.41(d,J＝8.7Hz,1H),5.58(d,J＝7.6Hz,1H),4.60(s,2H),4.50(d,J＝12.2Hz,2H),4.39(d,J＝4.9Hz,2H),4.36(s,1H),4.28–4.21(m,1H),4.1 9(d,J＝4.8Hz,2H),4.03(d,J＝12.3Hz,1H),3.19(t,J＝6.0Hz,2H),2.98(t,J＝16.9Hz,8H),2.76(d,J＝12.2Hz,1H),2.67(d,J＝4.6Hz,4H),2.60( d,J＝4.6Hz,1H),2.34(s,3H),2.11(dd,J＝8.8,4.7Hz,3H),1.73(s,4H),1.57(s,4H),1.35(s,2H),1.31(s,1H),1.27(s,4H).HRMS(ESI):calcd for C ₄₉ H ₅₉ ClFN ₁₁ O ₇ [M+H] ⁺ 968.4271,found 968.4332. Purity: 95.87% by HPLC (MeOH/H ₂ O = 80:20, t _R = 3.697 min).

Synthesis Route 10:

Example 20 was synthesized according to synthesis route 10.

Synthetic Route 10: Reagents and Conditions: (a) _K₂CO₃ , DMF, 60℃, 4h; (b _{) H₂} , Pd/C, EtOH, rt, 6h; (c) _NaHCO₃ , DMF, 80℃, 16h; (d) TFA, DCM, rt, 3h; (e) DIPEA, EDCI, HOBt, rt, 12h.

4-((1-(2-fluoro-4-nitrophenyl)piperidin-4-yl)methyl)piperazine-1-carboxylic acid tert-butyl ester (56b)

tert-butyl 4-((1-(2-fluoro-4-nitrophenyl)piperidin-4-yl)methyl)piperazine-1-carboxylate(56b)

Following general synthetic method 13, using 55 (500 mg, 3.14 mmol) as the reactant, the target product, white solid 56b (1.13 g, yield: 85.1%), was obtained. ESI-MS: m/z: [M+H] ⁺ 423.23.

4-((1-(4-amino-2-fluorophenyl)piperidin-4-yl)methyl)piperazine-1-carboxylic acid tert-butyl ester (57b)

tert-butyl 4-((1-(4-amino-2-fluorophenyl)piperidin-4-yl)methyl)piperazine-1-carboxylate(57b)

Following general synthesis method 2, using 56b (700 mg, 1.66 mmol) as the reactant, the target product, a white oily substance 57b (626 mg, yield: 96.3%), was obtained. ESI-MS: m/z: [M+H] ⁺ 393.26.

4-((1-(4-((2,6-dioxopiperidin-3-yl)amino)-2-fluorophenyl)piperidin-4-yl)methyl)piperazine-1-carboxylic acid tert-butyl ester (58b)

tert-butyl-4-((1-(4-((2,6-dioxopiperidin-3-yl)amino)-2-fluorophenyl)piperidin-4-yl)methyl)piperazine-1-carboxylate(58b)

Following general synthetic method 14, using 57b (700 mg, 1.27 mmol) as the reactant, the target product, dark purple solid 58b (301 mg, yield: 46.9%), was obtained. ESI-MS: m/z: [M+H] ⁺ 504.29.

3-((3-fluoro-4-(4-(piperazin-1-ylmethyl)piperidin-1-yl)phenyl)amino)piperidin-2,6-dione (59b)

3-((3-fluoro-4-(4-(piperazin-1-ylmethyl)piperidin-1-yl)phenyl)amino)piperidine-2,6-dione(59b)

Following general synthesis method 4, using 58b (283 mg, 0.56 mmol) as the reactant, the target product, a dark purple oil 59b (199 mg, yield: 87.8%), was obtained. ESI-MS: m/z: [M+H] ⁺ 404.24.

Example 20

2-((9-((5-chloro-2-(4-(4-((1-(4-(((2,6-dioxopiperidin-3-yl)amino)-2-fluorophenyl)piperidin-4-yl)methyl)piperazin-1-carbonyl)piperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-6-yl)oxy)-N-methylacetamide (Example 20)

2-((9-((5-chloro-2-(4-(4-((1-(4-((2,6-dioxopiperidin-3-yl)amino))-2-fluorophenyl)piperidin-4-yl)methyl)piperazine-1-carbonyl )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 (Example 20)

Following general synthesis method 9, using 35 (60 mg, 0.11 mmol) as the reactant, the target product, a brown solid, was obtained (Example 20, 11 mg, yield: 10.2%). ^¹H NMR (300 MHz, DMSO- _d6) was used. )δ10.80(s,1H),8.79(s,1H),8.06(s,1H),7.99(q,J＝3.7Hz,1H),7.58(d,J＝2.2 Hz,1H),7.40(d,J＝2.2Hz,1H),7.12(s,1H),6.84(t,J＝9.3Hz,1H),6.51(dd,J＝1 5.0,2.4Hz,1H),6.42(d,J＝8.9Hz,1H),5.81(d,J＝7.6Hz,1H),4.60(s,2H),4.50 (d,J＝12.6Hz,2H),4.40(t,J＝4.7Hz,2H),4.28(dd,J＝10.4,6.2Hz,1H),4.19(d,J ＝4.8Hz,2H),3.56(s,2H),3.46(s,2H),3.12(d,J＝10.8Hz,2H),2.96(t,J＝12.1H z,3H),2.67(d,J＝4.6Hz,3H),2.60(d,J＝4.7Hz,1H),2.55(s,4H),2.38(s,2H),2. 30(s,2H),2.20(d,J＝6.8Hz,2H),2.14–2.05(m,1H),1.78(d,J＝12.1Hz,2H),1.6 6(d,J＝12.6Hz,2H),1.51(t,J＝12.5Hz,2H),1.36–1.20(m,3H).HRMS(ESI):calcd for C ₄₅ H ₅₃ ClFN ₁₁ O ₇ [M+H] ⁺ 914.3802, found 914.3814.

Synthesis Route 11:

Example 21 was synthesized according to synthetic route 11.

Synthetic Route 11: Reagents and Conditions: (a) _Cs₂CO₃ , _Xantphos , Pd(OAc) _₂ , dioxane, 110℃, 10h; (b) LiOH, MeOH, _H₂O , rt, 12h; (c) EDCI, HOBt, TEA, DMF, 12h; (d) TFA, DCM, 3h; (e) NaBH(OAc) _₃ , DCE, 6h; (f) TFA, DCM, 3h; (g) EDCI, HOBt, DIPEA, DMF, 12h.

2-(3-fluoro-4-(methoxycarbonyl)phenyl)-2,8-diazaspiro[4.5]decane-8-carboxylic acid tert-butyl ester (63)

tert-butyl-2-(3-fluoro-4-(methoxycarbonyl)phenyl)-2,8-diazaspiro[4.5]decane-8-carboxylate(63)

Following general synthesis method 6, intermediate 62 (360 mg, 1.54 mmol) and 47b (482 mg, 1.70 mmol) were added to a sealed tube containing 10 mL of anhydrous dioxane solution. Cesium carbonate (1.01 g, 3.09 mmol), palladium acetate (17 mg, 0.08 mmol), and XantPhos (89 mg, 0.15 mmol) were then added sequentially. The mixture was purged with argon and reacted at 110 °C for 16 hours. The reaction was monitored by TLC until complete. After cooling to room temperature, the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by preparative chromatography (elution system: dichloromethane:methanol = 100:1, v/v) to obtain intermediate 63 as a white solid (390 mg, yield: 58.0%). ESI-MS: m/z: [M+H] ⁺ 393.21.

4-(8-(tert-Butoxycarbonyl)-2,8-diazaspiro[4.5]decane-2-yl)-2-fluorobenzoic acid (64)

4-(8-(tert-butoxycarbonyl)-2,8-diazaspiro[4.5]decan-2-yl)-2-fluorobenzoic acid(64)

Following general synthesis method 7, intermediate 63 (200 mg, 0.46 mmol) was added to a round-bottom flask containing a methanol:water mixture of 8 mL:2 mL, followed by lithium hydroxide (55 mg, 2.30 mmol). The reaction was carried out at 40 °C for 10 hours, and TLC was used to monitor the completeness of the reaction. After the reaction cooled to room temperature, dilute hydrochloric acid was added to adjust the pH of the solution to 5–6. The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (elution system: dichloromethane:methanol = 100:1, v/v) to obtain intermediate 64 as a white solid (180 mg, yield: 93.0%). ESI-MS: m/z: [M+H] ⁺ 379.20.

2-(4-((2,6-dioxopiperidin-3-yl)carbamoyl)-3-fluorophenyl)-2,8-diazaspiro[4.5]decane-8-carboxylic acid tert-butyl ester (65)

tert-butyl-2-(4-((2,6-dioxopiperidin-3-yl)carbamoyl)-3-fluorophenyl)-2,8-diazaspiro[4.5]decane-8-carboxylate(65)

Following general synthesis method 8, intermediate 64 (180 mg, 0.43 mmol) was added to a round-bottom flask containing 5 mL of LDM solution, followed by DIPEA (276 mg, 2.14 mmol), EDCI (123 mg, 0.64 mmol), and HOBt (87 mg, 0.64 mmol). The mixture was stirred at room temperature for half an hour, then 27a (83 mg, 0.51 mmol) was added, and the reaction was allowed to proceed overnight at room temperature. The reaction was monitored by TLC until complete. Extraction was performed with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (elution system: dichloromethane:methanol = 50:1, v/v) to give intermediate 65 as a white solid (160 mg, yield: 70.5%). ESI-MS: m/z: [M+H] ⁺ 489.24

N-(2,6-dioxopiperidin-3-yl)-2-fluoro-4-(2,8-diazaspiro[4.5]decane-2-yl)benzamide (66)

N-(2,6-dioxopiperidin-3-yl)-2-fluoro-4-(2,8-diazaspiro[4.5]decan-2-yl)benzamide(66)

Following general synthesis method 4, using 65 (320 mg, 0.65 mmol) as the reactant, the target product, a white oil 66 (237 mg, yield: 93.2%), was obtained. ESI-MS: m/z: [M+H] ⁺ 389.19.

4-((2-(4-((2,6-dioxopiperidin-3-yl)carbamoyl)-3-fluorophenyl)-2,8-diazaspiro[4.5]decane-8-yl)methyl)piperidin-1-carboxylic acid tert-butyl ester (67)

tert-butyl-4-((2-(4-((2,6-dioxopiperidin-3-yl)carbamoyl)-3-fluorophenyl)-2,8-diazaspiro[4.5]decan-8-yl)methyl)piperidine-1-carboxylate(67)

Following general synthetic method 1, using intermediate 66 (230 mg, 0.59 mmol) as the reactant, the target product, white solid 67 (195 mg, yield: 56.2%), was obtained. ESI-MS: m/z: [M+H] ⁺ 586.33.

N-(2,6-dioxopiperidin-3-yl)-2-fluoro-4-(8-(piperidin-4-ylmethyl)-2,8-diazaspiro[4.5]decane-2-yl)benzamide (68)

N-(2,6-dioxopiperidin-3-yl)-2-fluoro-4-(8-(piperidin-4-ylmethyl)-2,8-diazaspiro[4.5]decan-2-yl)benzamide(68)

Following general synthesis method 4, using 67 (190 mg, 0.32 mmol) as the reactant, the target product, a white oily substance 68 (143 mg, yield: 90.8%), was obtained. ESI-MS: m/z: [M+H] ⁺ 486.28.

Example 21

4-(8-((1-(1-(5-chloro-4-((6-(2-(methylamino)-2-oxoethoxy)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-9-yl)amino)pyrimidin-2-yl)piperidin-4-carbonyl)piperidin-4-yl)methyl)-2,8-diazaspiro[4.5]decane-2-yl)-N-(2,6-dioxopiperidin-3-yl)-2-fluorobenzamide (Example 21)

4-(8-((1-(1-(5-chloro-4-((6-(2-(methylamino)-2-oxoethoxy))-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-9-yl)amino)pyrimidin-2 -yl)piperidine-4-carbonyl)piperidin-4-yl)methyl)-2,8-diazaspiro[4.5]decan-2-yl)-N-(2,6-dioxopiperidin-3-yl)-2-fluorobenzamide (Example 21)

Following general synthesis method 9, using 35 (60 mg, 0.11 mmol) as the reactant, the target product, a brown solid, was obtained (Example 21, 17 mg, yield: 15.0%). ^¹H NMR (300 MHz, DMSO- _d6) )δ10.87(s,1H),8.79(s,1H),8.07(s,1H),8.00(q,J＝4.1,3.7Hz,1H),7.92(t, J＝7.8Hz,1H),7.66(t,J＝8.9Hz,1H),7.59(d,J＝2.3Hz,1H),7.40(d,J＝2.2Hz,1H ),7.12(s,1H),6.43(dd,J＝9.1,2.2Hz,1H),6.35(d,J＝15.1Hz,1H),4.74(dt,J＝ 12.4,6.4Hz,1H),4.60(s,2H),4.51(d,J＝12.0Hz,2H),4.41(d,J＝4.4Hz,2H),4. 33(s,1H),4.21–4.16(m,2H),4.03(d,J＝12.9Hz,1H),3.68–3.39(m,3H),3.17(s ,2H),2.99(dd,J＝24.0,11.5Hz,4H),2.83–2.72(m,1H),2.67(d,J＝4.6Hz,3H),2 .42–2.29(m,2H),2.15(d,J＝11.7Hz,2H),2.08–1.97(m,2H),1.86(s,3H),1.76( s,2H),1.68–1.49(m,8H),1.32(s,1H),1.26(d,J＝7.7Hz,4H).HRMS(ESI):calcd for C ₅₀ H ₅₉ ClFN ₁₁ O ₈ [M+H] ⁺ 996.4221, found 996.4289. Purity: 96.27% by HPLC (MeOH/H ₂ O = 80:20, t _R = 3.891 min).

Synthesis Route 12:

Example 22 was synthesized according to synthetic route 12.

Synthetic Route 12: Reagents and Conditions: (a) DIPEA, DMSO, 100℃, 10h; (b) Dys-Martin oxidant, DMF, 100℃, 2h; (c) NaBH(OAc) ₃ , DCE, 6h.

2-((9-((5-chloro-2-(4-(hydroxymethyl)piperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-6-yl)oxy)-N-methylacetamide (12b)

2-((9-((5-chloro-2-(4-(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(12b)

Intermediate 11 (300 mg, 0.69 mmol), starting material 11c (158 mg, 1.38 mmol), and DIPEA (151 mg, 1.17 mmol) were added to a round-bottom flask. 6 mL of DMSO solution was added, and the mixture was stirred overnight at 100 °C. TLC monitoring showed the reaction was complete. 40 mL of water was slowly added and stirred for ten minutes. The mixture was filtered, washed with water, and dried to obtain a yellow solid 12b (212 mg, yield: 59.9%). ESI-MS: m/z: [M+H] ⁺ 515.17.

2-((9-((5-chloro-2-(4-formylpiperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-6-yl)oxy)-N-methylacetamide (13b)

2-((9-((5-chloro-2-(4-formylpiperidin-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(13b)

Intermediate 12b (200 mg, 0.39 mmol) and Dysmart oxidant (496 mg, 1.17 mmol) were added to a round-bottom flask, followed by 5 mL of anhydrous DMF solution. The mixture was stirred at room temperature for 2 h, and the reaction was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give crude yellow solid 13b (141 mg, yield: 70.8%). ESI-MS: m/z: [M+H] ⁺ 513.16.

Example 22

2-((9-((5-chloro-2-(4-((4-((7-(4-(4-(2,6-dioxopiperidin-3-yl)-2-fluorophenyl)-2,7-diazaspiro[4.4]nonane-2-yl)methyl)piperidin-1-yl)methyl)piperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-6-yl)oxy)-N-methylacetamide (Example 22)

2-((9-((5-chloro-2-(4-((4-((7-(4-(2,6-dioxopiperidin-3-yl))-2-fluorophenyl)-2,7-diazaspiro[4.4]nonan-2-yl)methyl)piperidin-1-yl)m ethyl)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 (Example 22)

Intermediate 13b (80 mg, 0.16 mmol) and 54a (100 mg, 0.23 mmol) were added to a round-bottom flask containing 5 mL of anhydrous DMF solution. The mixture was stirred at room temperature for 15 minutes, then NaBH(OAc) _₃ (99 mg, 0.47 mmol) was added, and the mixture was stirred overnight at room temperature. The reaction was monitored by TLC until complete. The mixture was extracted three times with ethyl acetate, and the organic phases were combined. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by preparative chromatography (mobile phase: dichloromethane:methanol = 15:1, v/v) to give the target compound of Example 22 as a brown solid (10 mg, yield: 6.93%). ^¹H NMR (300 MHz, DMSO- _d₆) was performed. )δ9.21(s,1H),8.03(s,1H),7.79(s,1H),7.18(s,1H),7.07(d,J＝7.5Hz ,1H),6.77(d,J＝8.8Hz,1H),6.61(d,J＝2.9Hz,2H),6.59(d,J＝7.5Hz,1H) ,4.77(s,2H),4.41(dt,J＝12.5,7.1Hz,2H),4.39(s,2H),4.05(s,2H),3. 82(s,1H),3.71(dt,J＝9.5,7.1Hz,1H),3.66–3.51(m,3H),3.32(d,J＝9.5 Hz,1H),3.17–3.10(m,2H),2.81(s,3H),2.63–2.54(m,2H),2.59–2.46(m ,4H),2.33–2.19(m,2H),2.22–2.13(m,2H),2.00(d,J＝9.5Hz,1H),1.97– 1.86(m,2H),1.91–1.77(m,2H),1.82–1.69(m,2H),1.72–1.55(m,2H),1. 42–1.32(m,4H),1.39–1.30(m,2H),1.33–1.20(m,2H).HRMS(ESI):calcd for C ₄₈ H ₅₈ ClFN ₁₀ O ₆ [M+H] ⁺ 925.4213, found 925.4217. Purity: 96.02% by HPLC (MeOH/H ₂ O = 80:20, t _R = 3.797min).

Synthesis Route 13:

Example 23 was synthesized according to synthetic route 13.

Synthetic Route 13: Reagents and Conditions: (a _{) Na₂CO₃} , Pd(PPh) _₃Cl₂ , _dioxane , 110℃, 4h; (b) Pd/C, EtOH, rt, 16h; (c) LiOH, MeOH, _H₂O , rt, 16h; (d) BuLi, THF, DMF, -78℃, 6h; (e) _K₂CO₃ , DMF, rt, 5h; ( _f ) _NaBH₃CN , DIPEA, DCE; (g) TFA, DCM, 3h; (h) Dimethylchlorosilane, ACN, 6h; (i) Pd/C, _H₂ , rt, 12h; _Boc₂O , EtOH, THF, rt, 3h; ( _j ) _Tf₂O , TEA, DCM, rt, 3h; (k) _K₂CO₃ ,DMF,70℃,4h; (l)TFA,DCM,3h;(m)DIPEA,NMP,170℃,3h.

4-(2-fluoro-4-(methoxycarbonyl)phenyl)-3,6-dihydropyridine-1(2H)-carboxylic acid tert-butyl ester (70)

tert-butyl-4-(2-fluoro-4-(methoxycarbonyl)phenyl)-3,6-dihydropyridine-1(2H)-carboxylate(70)

Intermediates 69a (5.00 g, 21.46 mmol) and 69b (8.62 g, 27.89 mmol) were added to a sealed tube containing a solution of dioxane:water = 60:20 mL. Sodium carbonate (4.29 g, 42.92 mmol) and Pd(PPh) _3Cl2 (1.51 g, 2.15 mmol) were _then added sequentially. The mixture was purged with argon and reacted at 100 °C for 4 hours. The reaction was monitored by TLC until complete. After cooling to room temperature, the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (elution system: petroleum ether:ethyl acetate = 5:1, v/v) to give intermediate 70 as a white solid (4.6 g, yield: 63.9%). ESI-MS: m/z: [M+H] ⁺ 336.15.

4-(2-fluoro-4-(methoxycarbonyl)phenyl)piperidine-1-carboxylic acid tert-butyl ester (71)

tert-butyl 4-(2-fluoro-4-(methoxycarbonyl)phenyl)piperidine-1-carboxylate(71)

Following general synthesis method 2, using 70 (4.50 g, 13.42 mmol) as the reactant, the target product, a colorless oil 71 (4.32 g, yield: 95.4%), was obtained. ESI-MS: m/z: [M+H] ⁺ 338.17.

4-(1-(tert-Butoxycarbonyl)piperidin-4-yl)-3-fluorobenzoic acid (72)

4-(1-(tert-butoxycarbonyl)piperidin-4-yl)-3-fluorobenzoic acid(72)

Following general synthesis method 7, using 71 (4.40 g, 13.04 mmol) as the reactant, the target product, white solid 72 (4.06 g, yield: 96.3%), was obtained. ESI-MS: m/z: [M+H] ⁺ 324.15.

4-(4-fluoro-3-hydroxy-1-oxo-1,3-dihydroisobenzofuran-5-yl)piperidine-1-carboxylic acid tert-butyl ester (73)

tert-butyl-4-(4-fluoro-3-hydroxy-1-oxo-1,3-dihydroisobenzofuran-5-yl)piperidine-1-carboxylate(73)

Intermediate 72 (5.00 g, 15.46 mmol) was added to a double-necked flask containing a mixture of anhydrous THF and anhydrous DMF (40 mL:20 mL). The mixture was purged with argon and stirred at -78 °C for 30 min. Then, BuLi (23.19 mL, 23.19 mmol, 1 M in THF) was slowly added, and the reaction was carried out at room temperature for 6 h. The reaction was monitored by TLC until complete. After the reaction returned to room temperature, saturated ammonium chloride was slowly added to quench the reaction. The mixture was then extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Intermediate 73 was obtained as a white solid (2.40 g, yield: 44.2%) without further purification. ESI-MS: m/z: [M+H] ⁺ 352.15.

4-(2-fluoro-3-formyl-4-(methoxycarbonyl)phenyl)piperidine-1-carboxylic acid tert-butyl ester (74)

tert-butyl 4-(2-fluoro-3-formyl-4-(methoxycarbonyl)phenyl)piperidine-1-carboxylate(74)

Intermediate 73 (4.00 g, 11.38 mmol) was added to a round-bottom flask containing 40 mL of DMF solution, followed by potassium carbonate (3.15 g, 22.77 mmol). The reaction was carried out at room temperature for 4 hours, and TLC was used to monitor the completeness of the reaction. The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (elution system: petroleum ether: ethyl acetate = 5:1, v/v) to give intermediate 74 as a white solid (1.93 g, yield: 46.4%). ESI-MS: m/z: [M+H] ⁺ 366.16.

4-(2-fluoro-3-formyl-4-(methoxycarbonyl)phenyl)piperidine-1-carboxylic acid tert-butyl ester (75)

tert-butyl 4-(2-fluoro-3-formyl-4-(methoxycarbonyl)phenyl)piperidine-1-carboxylate(75)

Intermediate 74 (5.00 g, 13.68 mmol) was added to a round-bottom flask containing 60 mL of LCE solution, followed by DIPEA (3.54 g, 27.37 mmol) and 74a (2.10 g, 16.42 mmol). The mixture was stirred at room temperature for half an hour, then _NaBH3CN (1.72 g, 27.37 mmol) was added, and the reaction was carried out at room temperature for 6 hours. The reaction was monitored by TLC until complete. The mixture was concentrated under reduced pressure and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (elution system: dichloromethane:methanol = 100:1, v/v) to give intermediate 75 as a white solid (2.17 g, yield: 35.6%). ESI-MS: m/z: [M+H] ⁺ 446.20.

3-(4-fluoro-1-oxo-5-(piperidin-4-yl)isoindoline-2-yl)piperidin-2,6-dione (76)

3-(4-fluoro-1-oxo-5-(piperidin-4-yl)isoindolin-2-yl)piperidine-2,6-dione(76)

Following general synthesis method 4, using 75 (3.00 g, 6.73 mmol) as the reactant, the target product, a colorless oil 76 (2.19 g, yield: 94.16%), was obtained. ESI-MS: m/z: [M+H] ⁺ 336.15.

4-((1s,3s)-3-(benzyloxy)cyclobutoxy)piperidine-1-carboxylic acid benzyl ester (78)

benzyl 4-((1s,3s)-3-(benzyloxy)cyclobutoxy)piperidine-1-carboxylate(78)

Intermediate 77 (17.67 g, 75.74 mmol) was added to a round-bottom flask containing 100 mL of anhydrous acetonitrile solution and incubated on ice for 30 minutes. Then, dimethylchlorosilane (9.56 g, 100.99 mmol) and 77 (9.00 g, 50.50 mmol) were added sequentially, and the reaction was carried out at room temperature for 6 hours. The reaction was monitored by TLC until complete. The mixture was concentrated under reduced pressure and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (elution system: dichloromethane:methanol = 100:1, v/v) to give intermediate 78 as a white solid (8.20 g, yield: 41.1%). ESI-MS: m/z: [M+H] ⁺ 396.21.

4-((1s,3s)-3-hydroxycyclobutoxy)piperidine-1-carboxylic acid tert-butyl ester (79)

tert-butyl 4-((1s,3s)-3-hydroxycyclobutoxy)piperidine-1-carboxylate(79)

Intermediate 78 (8.00 g, 20.23 mmol) was added to a round-bottom flask containing 100 mL of methanol solution. Palladium on carbon (1.60 g, 20% m/m) was added, and the mixture was purged with hydrogen. The reaction was carried out overnight at room temperature, and TLC was used to monitor the completeness of the reaction. The mixture was filtered through diatomaceous earth, washed with methanol, and concentrated under reduced pressure to obtain a colorless oil. A 40 mL:40 mL mixture of ethanol and tetrahydrofuran was added, along with _Boc₂O (6.62 g, 30.34 mmol). The reaction was carried out for 4 h at room temperature, and TLC was used to monitor the completeness of the reaction. The mixture was extracted with ethyl acetate, and the organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (elution system: dichloromethane:methanol = 40:1, v/v) to obtain intermediate 79, a colorless oil (5.01 g, yield: 91.28%). ESI-MS: m/z: [M+H] ⁺ 272.18.

4-((1s,3s)-3-(((trifluoromethyl)sulfonyl)oxy)cyclobutoxy)piperidine-1-carboxylic acid tert-butyl ester (80)

tert-butyl-4-((1s,3s)-3-(((trifluoromethyl)sulfonyl)oxy)cyclobutoxy)piperidine-1-carboxylate(80)

Intermediate 79 (5.00 g, 18.43 mmol) was added to a round-bottom flask containing 50 mL of anhydrous DCM solution, followed by the addition of TEA (3.72 g, 36.86 mmol) and _Tf₂O (7.80 g, 27.64 mmol). The reaction was carried out at room temperature for 3 h, and the reaction was monitored by TLC until complete. The mixture was concentrated under reduced pressure and extracted with ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (elution system: dichloromethane:methanol = 50:1, v/v) to give intermediate 80 as a colorless oil (6.28 g, yield: 84.5%). ESI-MS: m/z: [M+H] ⁺ 404.13.

4-((1r, 3r)-3-(4-(2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxoisoindoline-5-yl)piperidin-1-yl)cyclobutoxy)piperidin-1-carboxylic acid tert-butyl ester (81)

tert-butyl-4-((1r,3r)-3-(4-(2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxoisoindolin-5-yl)piperidin-1-yl)cyclobutoxy)piperidine-1-carboxylate(81)

Intermediate 76 (1.00 g, 2.90 mmol) was added to a round-bottom flask containing 20 mL of anhydrous DMF solution, followed by the addition of potassium carbonate (0.80 g, 5.79 mmol) and 80 (1.28 g, 3.18 mmol). The reaction was carried out at 70 °C for 4 h, and the reaction was monitored by TLC until complete. Extraction was performed using ethyl acetate. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (elution system: dichloromethane:methanol = 50:1, v/v) to obtain intermediate 81 as a colorless oil (1.29 g, yield: 74.4%). ESI-MS: m/z: [M+H] ⁺ 599.32.

3-(4-fluoro-1-oxo-5-(1-((1r,3r)-3-(piperidin-4-yloxy)cyclobutyl)piperidin-4-yl)isoindoline-2-yl)piperidin-2,6-dione (82)

3-(4-fluoro-1-oxo-5-(1-((1r,3r)-3-(piperidin-4-yloxy)cyclobutyl)piperidin-4-yl)isoindolin-2-yl)piperidine-2,6-dione(82)

Following general synthesis method 4, using 81 (1.10 g, 1.84 mmol) as the reactant, the target product, a colorless oil 82 (0.85 g, yield: 92.8%), was obtained. ESI-MS: m/z: [M+H] ⁺ 499.26.

Example 23

2-((9-((5-chloro-2-(4-((1r,3r)-3-(4-(2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxoisoindoline-5-yl)piperidin-1-yl)cyclobutoxy)piperidin-1-yl)pyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-6-yl)oxy)-N-methylacetamide (Example 23)

2-((9-((5-chloro-2-(4-((1r,3r))-3-(4-(2-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-oxoisoindolin-5-yl)piperidin-1-yl)cyclobutoxy) 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 (Example 23)

Following general synthesis method 5, using 82 (171 mg, 0.34 mmol) as the reactant, the target product was obtained as a yellow solid (Example 23, 85 mg, yield: 41.3%). ^¹H NMR (300 MHz, DMSO- _d6) was performed. )δ8.61(s,1H),8.12(s,1H),7.83(s,1H),7.58(d,J＝7.5Hz,1H),6.62–6.62(m,2H),5.11(dt,J＝8.6,7.0Hz,2H),4.91(s,2 H),4.81(d,J＝19.4Hz,1H),4.50(m,J＝19.4Hz,1H),4.31(t,J＝4.6Hz,2H),4.22(m,J＝12.5,7.1,3.8Hz,2H),4.07(t,J＝4.7 Hz,2H),3.62(d,J＝12.6,7.1,3.9Hz,2H),3.32–3.21(m,3H),2.97(m,J＝14.3,7.1Hz,1H),2.92–2.81(m,2H),2.80(s,3H), 2.62–2.51(m,2H),2.51–2.36(m,2H),2.19–2.01(m,7H),1.91–1.72(m,4H),1.57(m,J＝13.7,7.1Hz,2H).HRMS(ESI):calcd for C ₄₅ H ₄₉ ClFN ₉ O ₈ [M+H] ⁺ 898.3377, found 898.3366. Purity: 96.33% by HPLC (MeOH/H ₂ O=80:20, t _R =4.147min).

Synthesis Route 14:

Example 24 was synthesized according to synthetic route 14.

Synthetic Route 14: Reagents and Conditions: (a) N,N-dimethyl-bromoacetamide, _Cs₂CO₃ , _DMF , rt, 3h; (b) Fe, _NH₄Cl , EtOH, _H₂O , 80℃, 3h; (c) DIPEA, DMSO, 100℃, 3h; (d) DIPEA, DMSO, 100℃, 10h; (e) DIPEA, EDCI, HOBt, rt, 12h.

N,N-Dimethyl-2-((9-nitro-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-6-yl)oxy)acetamide (8b)

N,N-dimethyl-2-((9-nitro-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-6-yl)oxy)acetamide(8b)

Intermediate 7 (1.00 g, 4.03 mmol) was added to a round-bottom flask, followed by 7b (0.83 g, 4.83 mmol) and cesium carbonate (2.63 g, 8.06 mmol), then DMF (15 mL). The mixture was stirred at room temperature for 4 h, and the reaction was monitored by TLC until it was complete. 50 mL of water was added, and the mixture was filtered, washed with water, and the filter cake was dried. Equal volumes of ethyl acetate and water were added to the filtrate for three hydration extractions. The organic layers were combined and concentrated under low pressure to remove the solvent. The residue was combined with the filter cake and purified by column chromatography (elution system: dichloromethane:methanol = 60:1, v/v) to obtain intermediate 8b as a yellow solid (0.53 g, yield: 37.3%). ¹ H NMR (300MHz, DMSO-d ₆ )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). ESI-MS:m/z:[M+H] ⁺ 334.10.

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

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

8b (0.40 g, 1.20 mmol) was added to a round-bottom flask containing anhydrous ethanol:water = 20 mL: 4 mL. Ammonium chloride (0.35 g, 6.26 mmol) was added, followed by reduced iron powder (0.34 g, 6.26 mmol) with stirring. The mixture was heated to 80 °C for 3 h, and the reaction was monitored by TLC until completion. Heating was removed, and the reaction solution was allowed to cool to room temperature. 2 mL of ammonia-methanol solution was added to adjust the pH of the solution to be greater than 7. The mixture was filtered through diatomaceous earth and washed with dichloromethane:methanol = 20:1 (1000 mL). The filtrate was collected and concentrated under low pressure to obtain a pale yellow solid 9b (0.23 g, yield: 63.5%). ESI-MS: m/z: [M+H] ⁺ 304.12.

2-((9-((2,5-dichloropyrimidin-4-yl)amino)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-6-yl)oxy)-N,N-dimethylacetamide (11b)

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(11b)

Intermediates 9b (0.40 g, 1.38 mmol), 10 (0.51 g, 2.77 mmol), and DIPEA (0.54 g, 4.02 mmol) were added to a round-bottom flask, followed by 8 mL of DMSO solution. The mixture was stirred at 100 °C for 3 h, and the reaction was monitored by TLC until complete. 35 mL of water was added, and the mixture was stirred for half an hour. The mixture was then filtered, washed with water, and dried to obtain a pale yellow solid, intermediate 11b (0.43 g, yield: 76.0%). ESI-MS: m/z: [M+H] ⁺ 450.06.

1-(5-chloro-4-((6-(2-(dimethylamino)-2-oxoethoxy)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinoline-9-yl)amino)pyrimidin-2-yl)piperidine-4-carboxylic acid (35b)

1-(5-chloro-4-((6-(2-(dimethylamino)-2-oxoethoxy)-5-oxo-2,3-dihydro-5H-[1,4]oxazino[2,3,4-ij]quinolin-9-yl)amino)pyrimidin-2-yl)piperidine-4-carboxylic acid(35b)

Intermediate 11b (300 mg, 0.69 mmol), starting material 12i (178 mg, 1.39 mmol), and DIPEA (151 mg, 1.17 mmol) were added to a round-bottom flask. 8 mL of DMSO solution was added, and the mixture was stirred at 100 °C for 10 h. TLC monitoring showed the reaction was complete. The pH of the solution was adjusted to 2–5 by slow dropwise addition of dilute hydrochloric acid, followed by the addition of 50 mL of water and stirring for ten minutes. The mixture was then filtered, washed with water, and dried to obtain a yellow solid 35b (310 mg, yield: 85.2%). ESI-MS: m/z: [M+H] ⁺ 543.17.

Example 24

2-((9-((5-chloro-2-(4-(4-((2-(2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindoline-5-yl)-2,8-diazaspiro[4.5]decane-8-yl)methyl)piperidin-1-carbonyl)piperidin-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 (Example 24)

2-((9-((5-chloro-2-(4-(4-((2-(2-(2,6-dioxopiperidin-3-yl))-1,3-dioxoisoindolin-5-yl)-2,8-diazaspiro[4.5]decan-8-yl)methyl)piperidine-1-c arbonyl)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,N-dimethylacetamide (Example 24)

Following general synthesis method 13, using 35b (60 mg, 0.10 mmol) as the reactant, the target product was obtained as a yellow solid (Example 24, 13 mg, yield: 12.2%). ^¹H NMR (300 MHz, DMSO- _d6) was used. )δ11.10(s,1H),8.79(s,1H),8.10–8.05(m,1H),8.00(d,J＝4.8Hz,1H),7.66 (d,J＝8.5Hz,1H),7.59(d,J＝2.2Hz,1H),7.40(d,J＝2.2Hz,1H),7.12(s,1H),6 .94(d,J＝2.1Hz,1H),6.83(dd,J＝8.7,2.1Hz,1H),5.08(dd,J＝12.6,5.4Hz,1H ),4.60(s,2H),4.51(d,J＝12.5Hz,2H),4.41(d,J＝4.5Hz,2H),4.37(s,1H),4. 19(t,J＝4.7Hz,2H),4.03(d,J＝12.7Hz,1H),3.49(s,3H),3.40(s,3H),3.31(s ,1H),3.10–2.85(m,6H),2.67(d,J＝4.6Hz,3H),2.62(s,1H),2.57(s,6H),2.4 0–2.12(m,2H),2.06–1.98(m,1H),1.94–1.87(m,2H),1.79(d,J＝13.8Hz,3H), 1.65(d,J＝12.0Hz,6H),1.51(d,J＝6.6Hz,2H),1.26(s,1H).HRMS(ESI):calcd for C ₅₂ H ₆₀ ClN ₁₁ O ₉ [M+H] ⁺ 1018.4264, found 1018.4270. Purity: 95.49% by HPLC (MeOH/H ₂ O = 80:20, t _R = 4.203 min).

Pharmacological activity evaluation

BCL6 protein degradation assay: OCI-LY1 cells (Zhejiang Meisen Cell Technology Co., Ltd.) were seeded in 6-well plates, and different concentrations of the compound were added. Twelve hours after drug administration, cells were collected by centrifugation, mixed with medium-efficiency RIPA lysis buffer (Beyotime Biotechnology), and centrifuged again. The supernatant was collected, and protein concentration was determined by the BCA method. Protein samples were mixed with protein loading buffer (Beyotime Biotechnology) and heated dry at 100°C for 10 min for sample preparation. The samples were then added to a 12% polyacrylamide gel SDS-PAGE. Electrophoresis was performed at 60V until the marker left the stacking gel, followed by continued electrophoresis at 120V. The membrane was transferred to a wet transfer buffer containing 10% methanol for 90 min. The transferred PVDF membrane was then cut into desired bands, blocked with milk for 2 hours, and anti-BCL6 antibody (abcam) and β-Actin antibody (Proteintech) diluted with milk were added to the corresponding bands, respectively, and incubated overnight at 4°C. The next day, the primary antibody was washed away with TBST, and the membrane was incubated with secondary antibody at room temperature for 45 min. The secondary antibody solution was then washed away again with TBST, and the membrane was scanned using an Odyssey Infrared Imaging System (LI-COR, Lincoln, Nebraska, USA). DC ₅₀ refers to the concentration of the BCL6 degrading agent required to achieve 50% degradation of BCL6 protein. The calculation process is as follows: Starting from 1000 nM, the degrading agent was serially diluted 5-fold to nine concentrations, and the degradation of BCL6 protein at each concentration was detected by Western blotting. Grayscale analysis was performed using ImageJ software to calculate the remaining amount of BCL6. The DC ₅₀ value was obtained by fitting the logarithm (Log(C)) of the remaining protein amount and concentration using Graphpad 8.0 software.

Table 1. Degradation of BCL6 in OCI-LY1 cells by the compounds.

Cell antiproliferative activity assay: Cells in logarithmic growth phase were seeded at a density of 5000 cells per well in 96-well plates (Coring, 3799) with 100 μL of RPMI 1640 medium (Adamas) containing 20% FBS and incubated overnight at 37°C in a 5% _CO2 incubator. The next day, 100 μL of different concentrations of drug solution prepared in the medium were added, with three replicates for each concentration (denoted as RLU _test ), and control and blank wells were also included. The control wells contained cells, culture medium, and the same concentration of drug solution (denoted as RLU _control ), while the blank wells contained culture medium (denoted as RLU _blank ). Nine days later, 100 μL of suspension cells were aspirated from each well and transferred to a 96-well black plate (Shanghai Wohong Biotechnology Co., Ltd., WHB-96-02). 100 μL of the CellTiter-Lumi luminescent cell viability assay kit (Beyotime Biotechnology) was added to each well, and the cells were incubated on a shaker in the dark for 10 min before detection. For adherent cells, 100 μL of culture medium was aspirated from each well and discarded. 100 μL of the assay kit was added to each well of the adherent cells, and the cells were lysed on a shaker for 2 min. The solution was then aspirated from each well and transferred to a 96-well black plate, incubated on a shaker in the dark for 8 min before detection. Chemiluminescence values were detected using the chemiluminescence module on a Thermo Scientific Varioskan Flash. The percentage of cell viability was calculated using the following formula: Cell viability (%) = (RLU _test - RLU _blank ) / (RLU _control - RLU _blank ) × 100%. The _IC50 value was calculated using Graphpad Prism 8.0 software.

Table 2. Antiproliferative activity of compounds against OCI-LY1 cells

Claims (10)

A compound with a chemical structure as shown in Formula I, or a pharmaceutically acceptable salt or solvate thereof:
POI-L-E3L
Ⅰ in: L is the Linker connecting POI and E3L; E3L is independently selected from:
in: _R1 , _R2 , _R3 , _R4 , _R5 , R6, _R7 , _R8 , R9, _R10 , _R11 , _R12 , _R13 , _R14 , _R15 , _R16 , _R17 , _R18 , _R19 , _R20 , _{and R21} are independently selected from -H, _halogen , _-C1 to _C3 alkyl, _-C1 to _C3 haloalkyl, and _-C1 to _C3 alkoxy. R ₂₂ is independently selected from -H, _-C1 to _-C3 alkyl groups; POIs are selected independently from:
in: R ₂₃ is independently selected from -H and halogens; _R24 and _R25 are independently selected from -H and _-C1 to _C3 alkyl groups; L is selected independently from:
in: n = 1, 2 or 3. A compound or a pharmaceutically acceptable salt or solvate thereof showing one of the following chemical structures:


A pharmaceutical composition characterized by comprising the compound of claim 1 or 2 or a pharmaceutically acceptable salt or solvate thereof. Use of the compound of claim 1 or 2 or a pharmaceutically acceptable salt or solvate thereof in the preparation of a BCL6 protein degrader. The use of the compound of claim 1 or 2 or a pharmaceutically acceptable salt or solvate thereof for the preparation of a medicament for treating a disease that is treated or alleviated by degradation of BCL6 protein. The use according to claim 5, wherein the disease is cancer. According to the use described in claim 6, the cancer is characterized in that: the cancer is Hodgkin lymphoma, B-cell non-Hodgkin lymphoma, T-cell non-Hodgkin lymphoma, NK/T-cell non-Hodgkin lymphoma, or diffuse large B-cell lymphoma. The use of the pharmaceutical composition of claim 3 for preparing a medicament for treating a disease, wherein the disease is a disease that is treated or alleviated by degrading BCL6 protein. The use according to claim 8, wherein the disease is cancer. According to the use described in claim 9, the cancer is characterized in that: the cancer is Hodgkin lymphoma, B-cell non-Hodgkin lymphoma, T-cell non-Hodgkin lymphoma, NK/T-cell non-Hodgkin lymphoma, or diffuse large B-cell lymphoma.