A type of indole ketone FLT3 protein degrading agent, its preparation method and medical use

Abstract

The present invention discloses a type of indole ketone FLT3 protein degrader, a preparation method thereof and a medical use thereof. The present invention relates to the field of pharmaceutical chemistry; the present invention discloses an indole ketone FLT3 protein degrader effective in vivo, which has a good therapeutic effect on acute myeloid leukemia, a tumor inhibition rate at the same dose is significantly better than that of FLT3 inhibitors, and has a good proliferation inhibition activity on acute myeloid leukemia cells resistant to FLT3 inhibitors, and has a good industrialization prospect.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical chemistry, and in particular to an indole ketone FLT3 protein degrading agent or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition containing these compounds, and their preparation methods and use in combating acute myeloid leukemia. Background Art

[0002] Abnormal activation of FMS-like receptor tyrosine kinase 3 (FLT3) is closely related to the occurrence, development and poor prognosis of acute myeloid leukemia (AML), and has become one of the important targets for targeted therapy of AML. Currently, a large number of FLT3 inhibitors have entered clinical research on AML and achieved good therapeutic effects, but the drug resistance problem after short-term use has seriously affected the efficacy of FLT3 inhibitors. Clinical studies have shown that ① secondary point mutations of FLT3, such as D835V / Y / F / H, F691I / L, Y842C / H, A848P and A627P, can lead to resistance to FLT3 inhibitors in patients; ② increased expression of FLT3 and increased concentration of FLT3 ligands in the bone marrow microenvironment promote the combination of the two, reactivating the FLT3 signaling pathway and leading to resistance; ③ CYP3A4 in bone marrow stromal cells rapidly metabolizes free FLT3 inhibitors, reducing drug concentrations and leading to resistance; ④ bypass activation of other signaling pathways is also an important factor in drug resistance. Therefore, resistance to FLT3 inhibitors in AML patients is the result of the combined action of multiple factors. Traditional reversible FLT3 inhibitors are difficult to achieve ideal therapeutic effects. There is an urgent need to develop an anti-AML drug with a new mechanism of action to fundamentally solve the problem of drug resistance in the treatment of AML with existing drugs.

[0003] Proteolytic targeting chimera (PROTAC) technology is a new treatment strategy that uses chemical small molecules to induce ubiquitination of target proteins and then degrade them through the proteasome. It is an emerging direction in the field of drug research and development in recent years. Compared with traditional small molecule inhibitors that inhibit the function of target proteins through the "occupancy-driven" mode of action to treat diseases, PROTAC is an "event-driven" transient mode of action. It does not affect the function of proteins, but mediates the degradation of pathogenic target proteins. As long as PROTAC mediates the formation of a ternary complex and labels the target protein with ubiquitination, it can theoretically be recycled and reused. Therefore, the catalyst amount can play a role, avoiding the drug resistance caused by long-term and high-dose administration leading to increased target protein expression, target protein mutations, and bypass signal activation. Therefore, the development of FLT3-PROTACs that target the degradation of FLT3 protein is expected to solve the drug resistance caused by FLT3 mutations or high expression from the source.

[0004] In 2018, the Crews team reported a FLT3 protein degrader based on Quizartinib [J AmChem Soc, 2018, 140, 16428-16432], and in 2021, the Yang team reported a FLT3 protein degrader based on Dovitinib [J Med Chem, 2021, 64, 16497-16511]. These two PROTACs molecules can effectively degrade FLT3 protein in MV4-11 cells, and significantly enhance the anti-proliferative activity of AML cells carrying FLT3-ITD mutations. Unfortunately, these two FLT3 degraders have no relevant in vivo efficacy data, and no relevant anti-resistance studies have been conducted. Practice has proved that the development of FLT3 degraders is feasible, but the problem of drugability has always been a problem restricting the development of PROTACs. At present, no in vivo effective FLT3 degraders have entered the clinical research stage, and the existing FLT3 degraders have serious deficiencies in in vivo efficacy and drugability. Therefore, how to design and synthesize an effective FLT3 degrader in vivo and overcome the drug resistance of traditional FLT3 inhibitors remains a key scientific issue that urgently needs to be solved in the field of AML treatment.

[0005]

[0006] Based on the previously discovered FLT3 inhibitor candidate compound PX-A13, the present invention uses thalidomide as the ligand of the E3 ligase CRBN, and designs and synthesizes a series of PROTACs that target the degradation of FLT3 protein by optimizing the type, length, and connection mode of the Linker. Through systematic biological activity and drugability evaluation, a new type of FLT3 protein degrader with effective in vivo and significant anti-AML effect was successfully discovered. Summary of the invention

[0007] In view of the above problems, the present invention provides an indole one FLT3 protein degrading agent represented by general formula (I) or a pharmaceutically acceptable salt thereof.

[0008] The technical solution of the present invention is: the present invention discloses an indole ketone FLT3 protein degrading agent or a pharmaceutically acceptable salt, polymorph or solvate thereof, characterized in that the chemical structural formula of the derivative is as shown in formula (I):

[0009]

[0010] in:

[0011] R 1 One or more selected from hydrogen atom, halogen, alkyl, alkoxy, haloalkyl or amino;

[0012] R 2 is selected from a hydrogen atom, a methyl group or a halomethyl group;

[0013] R 3 is selected from a hydrogen atom or a halogen;

[0014] R 4 Selected from a hydrogen atom or forming a carbonyl group with an adjacent carbon;

[0015] X is selected from -NHCO-, -CONH- or -NH-;

[0016] m is 0-6;

[0017] Linker is selected from

[0018] n is 1-12;

[0019] Preferred typical compounds of the present invention are as follows, but not limited to:

[0020]

[0021]

[0022]

[0023]

[0024] One object of the present invention is to provide a preparation method; the specific preparation steps are as follows:

[0025] In the process of preparing compound III by reacting compound II with a flexible fatty chain or polyethylene glycol linker, the required acid binding agent is selected from triethylamine, N,N-diisopropylethylamine (DIPEA), 4-dimethylaminopyridine (DMAP), pyridine, sodium acetate, sodium carbonate or potassium carbonate, preferably DIPEA; the reaction solvent is selected from N,N-dimethylformamide (DMF), N,N-dimethylacetamide (DMA) or dimethyl sulfoxide (DMSO), preferably DMA; the reaction temperature is 70°C to 100°C, preferably 80°C.

[0026] The process of preparing compound I by reacting compound III with compound IV, wherein the condensing agent used is selected from 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU), hexafluorophosphate benzotriazol-1-yl-oxytripyrrolidinophosphine (PyBOP), 1-hydroxybenzotriazole (HOBT) / 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI), dicyclohexylcarbodiimide (DCC) or N,N' -carbonyldiimidazole (CDI), preferably PyBOP; the acid-binding agent is selected from triethylamine, N,N-diisopropylethylamine (DIPEA), 4-dimethylaminopyridine (DMAP), pyridine, sodium acetate, sodium carbonate or potassium carbonate, preferably DIPEA; the reaction solvent is selected from N,N-dimethylformamide (DMF), N,N-dimethylacetamide (DMA) or dimethyl sulfoxide (DMSO), preferably DMF; the reaction temperature is 10°C to 50°C, preferably 20°C to 30°C.

[0027] The process of preparing compound V by reacting compound IV with a rigid heterocyclic linker is divided into two steps. The first step is to react an amine with a carbonyl group under acidic conditions to form a Schiff base, and the acid used is selected from acetic acid (AcOH), formic acid (HCOOH), p-toluenesulfonic acid (PTSA), preferably AcOH; the reducing agent used in the second step is selected from sodium triacetoxyborohydride (STAB), sodium cyanoborohydride, sodium borohydride, preferably STAB; the reaction solvent is selected from methanol, dichloromethane (DCM), tetrahydrofuran (THF), dimethyl sulfoxide (DMSO) or a mixed solvent of any two to three of them, preferably DMSO; the reaction temperature is 10°C to 50°C, preferably 20°C to 30°C; the acid used to remove the Boc protecting group is selected from hydrochloric acid, trifluoroacetic acid, sulfuric acid, p-toluenesulfonic acid, preferably hydrochloric acid; the reaction solvent is selected from ethyl acetate, methanol, dichloromethane, acetone, dioxane or a mixed solvent of any of them, preferably ethyl acetate; the reaction temperature is 10°C to 40°C, preferably 20°C.

[0028] In the process of preparing compound I by reacting compound V with compound VI, the required acid binding agent is selected from triethylamine, N,N-diisopropylethylamine (DIPEA), 4-dimethylaminopyridine (DMAP), pyridine, sodium acetate, sodium carbonate or potassium carbonate, preferably DIPEA; the reaction solvent is selected from N,N-dimethylformamide (DMF), N,N-dimethylacetamide (DMA) or dimethyl sulfoxide (DMSO), preferably DMSO; the reaction temperature is 70°C to 100°C, preferably 80°C.

[0029] The process of preparing compound I·A from compound I by salt formation, wherein reactant A is hydrogen chloride, sulfuric acid, phosphoric acid, carbonic acid, oxalic acid, citric acid, succinic acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, preferably hydrogen chloride; and the solvent is methanol, ethanol, dichloromethane, acetone, ethyl acetate, toluene, tetrahydrofuran, or a mixed solvent of any of them, preferably ethyl acetate.

[0030] The second object of the present invention is to provide a pharmaceutical composition, comprising the indolinone FLT3 protein degrader, a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier, additive, adjuvant or excipient.

[0031] Another object of the present invention is to provide the use of the indolinone FLT3 protein degrader or pharmaceutically acceptable salt, and pharmaceutical composition thereof in the preparation of FLT3 protein degrader; the compound or pharmaceutically acceptable salt thereof, and pharmaceutical composition thereof of the present invention have strong degradation activity on FLT3, and thus can be used to prepare drugs for treating related tumor diseases caused by abnormal FLT3 expression.

[0032] Another object of the present invention is to provide the use of the indolinone FLT3 protein degrader or pharmaceutically acceptable salt thereof, and the pharmaceutical composition thereof in the preparation of a drug for treating acute myeloid leukemia. In vitro anti-tumor activity experiments show that the compounds of the present invention can significantly inhibit the proliferation of acute myeloid leukemia cells; whole animal experiments show that the compounds of the present invention have a good therapeutic effect on acute myeloid leukemia; therefore, the compounds of the present invention or their pharmaceutically acceptable salts, and the pharmaceutical composition thereof can be used as a single therapeutic agent, or in combination with other anti-tumor drugs, for the treatment of acute myeloid leukemia.

[0033] Another object of the present invention is to provide the use of the indolinone FLT3 protein degrader or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition thereof in the preparation of a drug for treating acute myeloid leukemia resistant to FLT3 inhibitors; in vitro antitumor activity experiments show that the compounds of the present invention have a significant antiproliferative effect on acute myeloid leukemia cells resistant to FLT3 inhibitors; therefore, the compounds of the present invention or a pharmaceutically acceptable salt thereof, and a pharmaceutical composition thereof can be used as a single therapeutic agent, or in combination with other antitumor drugs, for the treatment of acute myeloid leukemia resistant to FLT3 inhibitors.

[0034] The beneficial effects of the present invention are as follows: the present invention discloses an orally effective indole one FLT3 protein degrader, which has a good therapeutic effect on acute myeloid leukemia, and the tumor inhibition rate at the same dose is significantly better than that of FLT3 inhibitors, and has a good proliferation inhibition activity on acute myeloid leukemia cells resistant to FLT3 inhibitors, can overcome the resistance of FLT3 inhibitors, and has a good industrialization prospect. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 It is a preparation flow chart of the present invention;

[0036] Figure 2 It is a comparison diagram of the degradation effect of compounds A1-A29 in Example 32 of the present invention on FLT3 protein in MV-4-11 and MOLM-13 cells; wherein, (a) is a schematic diagram of the degradation effect of compounds A1-A29 on FLT3 protein in MV-4-11; (b) is a schematic diagram of the degradation effect of compounds A1-A29 on FLT3 protein in MOLM-13 cells. DETAILED DESCRIPTION

[0037] In order to more clearly illustrate the technical solution of the present invention, the technical solution of the present invention is further described in detail below:

[0038] Example 1

[0039] Synthesis of (Z)-2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)oxy)-N-(2-(3-(4-(2-((5-((5-fluoro-2-oxoindole-3-ylidene)methyl)-4-methyl-1H-pyrrol-3-yl)amino)-2-oxoethyl)piperazin-1-yl)-3-oxopropoxy)ethyl)acetamide (A01):

[0040]

[0041] Synthesis of intermediate II-1: tert-Butyl 2-hydroxyacetate (0.51 g, 3.85 mmol) and triphenylphosphine (1.35 g, 5.15 mmol) were dissolved in 10 mL of anhydrous THF. After the reaction system was cooled to 0°C, a solution of DTBAD (1.15 g, 5.15 mmol) in anhydrous THF was added, and then a solution of compound Ⅰ-1 (1.1 g, 3.85 mmol) in anhydrous THF was added. The reaction system was stirred at 0°C for 1 h, then returned to room temperature and reacted overnight. The reaction was monitored by LC-MS to be complete. After the solvent was removed by vortexing, it was dissolved in 10 mL of dichloromethane, and then 13 mL of formic acid was added. The system was heated to 40°C for 36 h. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 1:1). The reaction was completed, concentrated under reduced pressure, and then separated by column chromatography to obtain a white solid powder (700 mg, yield: 60%).

[0042]

[0043] Synthesis of intermediate III-1: Compound II-1 (0.2 g, 0.63 mmol) and linker (0.63 mmol) were dissolved in 5 mL of N,N-dimethylformamide, and HATU (0.23 g, 0.98 mmol) and DIPEA (0.3 mL, 1.26 mmol) were added. The reaction system was reacted at room temperature for 2 h. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 1:1) to complete. The reaction was poured into water, extracted with ethyl acetate (30 mL × 3), the organic phases were combined, washed with water (30 mL × 2), washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo. Then 5 mL of dichloromethane was added thereto, and 4 ml of formic acid was added. The reaction system was then heated to 40 ° C and acid-hydrolyzed at this temperature until the reaction was complete. After the reaction was completed, the reaction was poured into water, extracted with ethyl acetate (30 mL × 2), the organic phases were combined, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product, which was directly used for the next step reaction.

[0044]

[0045] Synthesis of intermediate V: Compound IV (0.26 g, 0.78 mmol) and 1-tert-butyloxycarbonylpiperazine (0.79 g, 3.9 mmol) were dissolved in N,N-dimethylformamide (10 mL), and the temperature was raised to 50 °C for reaction for 12 h. The reaction was completed after LC-MS monitoring. 30 mL of water was added to the reaction system, and a solid precipitated, namely the product, which was filtered and dried to obtain a brown-yellow solid V (0.37 g, yield: 90%). 1HNMR(500MHz,DMSO-d6)δ13.45(s,1H),10.89(s,1H),9.22(s,1H),7.77(dd,J=9.4,2.6Hz,1H),7.74(s,1H),7.64(d,J=3.0Hz,1H) ,6.96–6.91(m,1H),6.84(dd,J=8.4,4.5Hz,1H),3.39(t,J=4.9Hz,4H),3.17(s,2H),2.50–2.41(m,4H),2.28(s,3H),1.41(s,9H).

[0046]

[0047] Synthesis of intermediate VI: Compound V (0.35 g, 0.72 mmol) was dissolved in a small amount of a mixture of dichloromethane and methanol, and then a hydrochloric acid ethyl acetate solution was added, and the mixture was sealed and reacted at room temperature. The reaction was monitored by TLC, and the solvent was removed by vacuum concentration, and then stirred with ethyl acetate (10 mL) for 0.5 h, and the ethyl acetate was removed by vacuum concentration. Finally, a large amount of ethyl acetate was used for slurrying, and the filter cake was washed with a large amount of ethyl acetate, and then washed with a small amount of n-hexane, and the filter cake was dried by oil pump. A brown-yellow compound VI (0.26 g, yield: 85%) was obtained. 1 H NMR (500MHz, DMSO-d6) δ13.54(s,1H),10.96(s,1H),9.27(s,1H),7.77(dd,J=9.3,2.6Hz,1H),7.73(s,1H),7.62(d,J=3.0Hz,1H), 6.94(ddd,J=9.5,8.4,2.6Hz,1H),6.82(dd,J=8.4,4.5Hz,1H),3.43(t,J=4.8Hz,6H),3.21(s,2H),2.50–2.41(m,4H),2.23(s,3H).

[0048]

[0049] Synthesis of compound A01: Compound III-1 (110 mg, 0.25 mmol), compound VI (103 mg, 0.25 mmol) and PyBOP (195 mg, 0.375 mmol) were dissolved in 5 mL of N,N-dimethylformamide, and DIPEA (180 μL, 1.0 mmol) was added under stirring. The system was reacted at room temperature for 2 h. The reaction was completed under LC-MS monitoring. 15 mL of water was added to the reaction system to precipitate solids, which were filtered and dried to obtain a crude product. The crude product was separated by column chromatography (DCM:MeOH=100:1~30:1) to obtain pure brown-yellow solid powder A01 (47 mg, yield: 23%). 1 H NMR (500MHz, DMSO-d6) δ13.45(s,1H),11.13(s,1H),10.89(s,1H),9.24(s,1H),8.01(t,J=5.7Hz,1H),7.81(t,J=7.9Hz,1H),7.77 (dd,J=9.4,2.6Hz,1H),7.73(s,1H),7.64(d,J=3.1Hz,1H),7.50(d,J=7.3Hz,1H),7.41(d,J=8.4Hz,1H),6.93(t,J=9.1Hz,1H),6. 84(dd,J=8.5,4.5Hz,1H),5.12(dd,J=12.8,5.4Hz,1H),4.80(s,2H),3.64(t,J=6.6Hz,2H),3.56–3.47(m,4H),3.46(t,J=5.7Hz,2 H),3.31(t,J=15.0Hz,2H),3.17(s,2H),2.94–2.86(m,1H),2.65–2.61(m,1H),2.60–2.52(m,6H),2.28(s,3H),2.09–2.03(m,2H). 13 C NMR (126MHz, DMSO-d6) δ173.23,170.34,169.86,169.23,167.36,167.19,165.93,159. 58,157.71,155.46,140.06,137.41,135.17,133.49,125.74,125.71,125.65,120.85,1 19.52,117.24,116.52,116.52,115.73,115.29,111.00,110.41,69.05,68.02,67.01,6 3.91,62.93,52.89,52.87,49.28,38.83,33.19,31.42,22.47,9.11.HR-MS(ESI):calcd for C40 H 42 F8O 10 [M+H] + :813.3002; found:813.3009.

[0050] Example 2

[0051] Synthesis of (Z)-2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)oxy)-N-(2-(3-(4-(2-((5-((5-fluoro-2-oxoindole-3-ylidene)methyl)-4-methyl-1H-pyrrol-3-yl)amino)-2-oxoethyl)piperazin-1-yl)-3-oxopropoxy)ethyl)acetamide (A02)

[0052]

[0053] Synthesis of intermediate III-2: Compound II-1 (0.2 g, 0.63 mmol) and linker (0.63 mmol) were dissolved in 5 mL of N,N-dimethylformamide, and HATU (0.23 g, 0.98 mmol) and DIPEA (0.3 mL, 1.26 mmol) were added. The reaction system was reacted at room temperature for 2 h. The reaction was monitored by TLC (petroleum ether: ethyl acetate = 1:1) until completion. The reaction was poured into water, extracted with ethyl acetate (30 mL × 3), the organic phases were combined, washed with water (30 mL × 2), washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo. Then 5 mL of dichloromethane was added thereto, and 4 ml of formic acid was added. The reaction system was then heated to 40 ° C and acidolyzed at this temperature until the reaction was complete. After the reaction was completed, the reaction was poured into water, extracted with ethyl acetate (30 mL × 2), the organic phases were combined, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product, which was directly used for the next step reaction.

[0054]

[0055] Synthesis of compound A02: Compound III-2 (120 mg, 0.25 mmol), compound VI (103 mg, 0.25 mmol) and PyBOP (195 mg, 0.375 mmol) were dissolved in 5 mL of N,N-dimethylformamide, and DIPEA (180 μL, 1.0 mmol) was added under stirring. The system was reacted at room temperature for 2 h. The reaction was completed under LC-MS monitoring. 15 mL of water was added to the reaction system to precipitate solids, which were filtered and dried to obtain a crude product. The crude product was separated by column chromatography (DCM:MeOH=100:1~30:1) to obtain pure brown-yellow solid powder A02 (56 mg, yield: 26%). 11H NMR (500 MHz, DMSO-d6) δ 13.45 (s, 1H), 11.12 (s, 1H), 10.88 (s, 1H), 9.24 (s, 1H), 8.00 (t, J = 5.6 Hz, 1H), 7.81 (t, J = 7.9 Hz, 1H), 7.76 (dd, J = 9.4, 2.6 Hz, 1H), 7.73 (s, 1H), 7.63 (d, J = 3.1 Hz, 1H), 7.50 (d, J = 7.3 Hz, 1H), 7.41 (d, J = 8.5 Hz, 1H), 6.93 (td, J = 9.4, 9.0, 2.6 Hz, 1H), 6.84 (dd, J = 8.4, 4.5 Hz, 1H), 5.12 (dd, J = 12.7, 5.4 Hz, 1H), 4.80 (s, 2H), 3.63 (t, J = 6.6 Hz, 2H), 3.48 (s, 8H), 3.47 (d, J = 5.7 Hz, 2H), 3.32 (d, J = 5.6 Hz, 2H), 3.17 (s, 2H), 2.94–2.86 (m, 1H), 2.62 (s, 1H), 2.60–2.52 (m, 6H), 2.28 (s, 3H), 2.09–1.99 (m, 2H). 13 13C NMR (126 MHz, DMSO-d6) δ 173.22, 170.32, 169.86, 169.22, 167.67, 167.35, 167.18, 165.90, 157.72, 155.42, 137.39, 135.17, 133.50, 127.58, 127.51, 125.70, 125.66, 124.28, 121.22, 120.82, 119.52, 117.24, 116.51, 115.70, 112.81, 110.47, 106.50, 106.30, 70.07 (2C), 69.29, 67.99, 67.24, 61.13, 53.40, 52.93, 49.29 (2C), 45.52, 41.52, 33.22, 31.42, 22.47, 9.10. HR-MS (ESI): calcd for C 42 H 46 FN8O 11 [M + H]+: 857.3265; found: 857.3265.

[0056] Example 3

[0057] Synthesis of (Z)-2-(4-(3-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)amino)ethoxy)propionyl)piperazin-1-yl)-N-(5-(5-fluoro-2-oxoindole-3-ylidene)methyl)-4-methyl-1H-pyrrol-3-yl)acetamide (A03)

[0058]

[0059] Synthesis of intermediate II-2: I-2 (200 mg, 0.72 mmol) and polyethylene glycol linker (0.66 mmol) were dissolved in DMA 5 mL, and DIPEA (300 μL, 1.72 mmol) was added. The reaction system was heated to 80 ° C. and reacted under this condition for 4 h. After the reaction was completed, the temperature was lowered to room temperature, and the reaction solution was poured into water (20 mL), extracted with ethyl acetate (30 mL × 2), the organic phases were combined, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to remove the solvent. Then 5 mL of dichloromethane was added, and 4 mL of formic acid was added. The reaction system was heated to 40 ° C. and acid hydrolyzed at this temperature until the reaction was complete. After the reaction was completed, the reaction was poured into water, extracted with ethyl acetate (30 mL × 2), the organic phases were combined, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product, which was directly used for the next step reaction.

[0060]

[0061] Synthesis of compound A03: Compound II-2 (110 mg, 0.25 mmol), compound VI (103 mg, 0.25 mmol) and PyBOP (195 mg, 0.375 mmol) were dissolved in 5 mL of N,N-dimethylformamide, and DIPEA (180 μL, 1.0 mmol) was added under stirring. The system was reacted at room temperature for 2 h. The reaction was completed under LC-MS monitoring. 15 mL of water was added to the reaction system to precipitate solids, which were filtered and dried to obtain a crude product. The crude product was separated by column chromatography (DCM:MeOH=100:1~30:1) to obtain pure brown-yellow solid powder A03 (63 mg, yield: 31%). 1H NMR (500MHz, DMSO-d6) δ13.45(s,1H),11.12(s,1H),10.88(s,1H),9.24(s,1H),7.76(dd,J=9.4,2.6Hz,1H),7.73(s,1H),7.64( d,J=3.0Hz,1H),7.58(t,J=7.9Hz,1H),7.15(d,J=8.4Hz,1H),7.04(d,J=7.1Hz,1H),6.93(t,J=7.7Hz,1H),6.84(dd,J=8.5,4.4H z,1H),6.61(t,J=5.9Hz,1H),5.06(dd,J=12.7,5.4Hz,1H),3.64–3.62(m,4H),3.57–3.56(m,2H),3.53–3.51(m,6H),3.48–3.45( m,2H),3.17(s,2H),2.94–2.86(m,1H),,2.61(s,1H),2.55(t,J=18.5Hz,4H),2.48–2.45(m,2H),2.28(s,3H),2.06–1.99(m,2H). 13 C NMR(126MHz,DMSO-d6)δ173.26,170.54,169.87,169.40,169.20,167.74,167.64,157.7 2,146.87,136.67,135.17,132.55,127.59,125.70,125.66,124.29,121.20,119.51,11 7.91,115.73,113.01,112.82,111.13,109.72,107.24,70.17(2C),69.36,67.31,61.13 ,60.22,53.39,52.93,49.03(2C),45.49,33.23,31.46,22.61,9.10.HR-MS(ESI):calcd for C 40 H 44 FN8O9[M+H] + :799.3210; found:799.3195.

[0062] Example 4

[0063] Synthesis of (Z)-2-(4-(3-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)amino)ethoxy)propionyl)piperazin-1-yl)-N-(5-(5-fluoro-2-oxoindole-3-ylidene)methyl)-4-methyl-1H-pyrrol-3-yl)acetamide (A04)

[0064]

[0065] Synthesis of intermediate II-3: I-2 (200 mg, 0.72 mmol) and polyethylene glycol linker (0.66 mmol) were dissolved in DMA 5 mL, and DIPEA (300 μL, 1.72 mmol) was added. The reaction system was heated to 80 ° C. and reacted under this condition for 4 h. After the reaction was completed, the temperature was lowered to room temperature. The reaction solution was poured into water (20 mL), extracted with ethyl acetate (30 mL × 2), the organic phases were combined, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to remove the solvent. Then 5 mL of dichloromethane was added, and 4 mL of formic acid was added. The reaction system was heated to 40 ° C. and acid hydrolyzed at this temperature until the reaction was complete. After the reaction was completed, the reaction was poured into water, extracted with ethyl acetate (30 mL × 2), the organic phases were combined, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product, which was directly used for the next step reaction.

[0066]

[0067] Synthesis of compound A04: Compound II-3 (75 mg, 0.19 mmol), compound VI (81 mg, 0.19 mmol) and PyBOP (150 mg, 0.29 mmol) were dissolved in 5 mL of N,N-dimethylformamide, and DIPEA (140 μL, 0.77 mmol) was added under stirring. The system was reacted at room temperature for 2 h. The reaction was completed under LC-MS monitoring. 15 mL of water was added to the reaction system to precipitate solids, which were filtered and dried to obtain a crude product. The crude product was separated by column chromatography (DCM:MeOH=100:1~30:1) to obtain pure brown-yellow solid powder A04 (44 mg, yield: 31%). 11H NMR (500 MHz, DMSO-d6) δ 13.45 (s, 1H), 11.11 (s, 1H), 10.89 (s, 1H), 9.23 (s, 1H), 7.76 (dd, J = 9.4, 2.6 Hz, 1H), 7.73 (s, 1H), 7.64 (d, J = 3.1 Hz, 1H), 7.59 (t, J = 8.0 Hz, 1H), 7.16 (d, J = 8.6 Hz, 1H), 7.05 (d, J = 7.0 Hz, 1H), 6.93 (td, J = 9.4, 9.0, 2.6 Hz, 1H), 6.84 (dd, J = 8.4, 4.5 Hz, 1H), 6.59 (t, J = 5.8 Hz, 1H), 5.06 (dd, J = 12.7, 5.4 Hz, 1H), 3.69 (t, J = 6.6 Hz, 2H), 3.61 (t, J = 5.4 Hz, 2H), 3.55–3.50 (m, 4H), 3.47 (t, J = 5.5 Hz, 2H), 3.16 (s, 2H), 2.94–2.86 (m, 1H), 2.61 (s, 1H), 2.60–2.53 (m, 4H), 2.50–2.46 (m, 2H), 2.28 (s, 3H), 2.06–1.99 (m, 2H). 13 13C NMR (126 MHz, DMSO-d6) δ 173.25, 170.51, 169.87, 169.40, 169.11, 167.73, 167.68, 157.72, 146.87, 136.70, 135.17, 132.57, 127.59, 125.70, 125.66, 124.28, 121.20, 119.51, 117.89, 115.73, 113.01, 111.15, 110.40, 109.73, 106.51, 69.12, 67.17, 61.11, 60.22, 53.41, 52.92, 49.04 (2C), 42.15, 33.20, 31.45, 22.63, 9.09. HR-MS (ESI): calcd for C 38 H 40 FN8O8 [M + H] + : 755.2948; found: 755.2951.

[0068] Example 5

[0069] Synthesis of (Z)-2-(4-(3-(2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)amino)ethoxy)ethoxy)propionyl)piperazin-1-yl)-N-(5-(5-fluoro-2-oxoindole-3-ylidene)methyl)-4-methyl-1H-pyrrol-3-yl)acetamide (A05)

[0070]

[0071] Synthesis of intermediate II-4: I-2 (200 mg, 0.72 mmol) and polyethylene glycol linker (0.66 mmol) were dissolved in DMA 5 mL, and DIPEA (300 μL, 1.72 mmol) was added. The reaction system was heated to 80 ° C. and reacted under this condition for 4 h. After the reaction was completed, the temperature was lowered to room temperature. The reaction solution was poured into water (20 mL), extracted with ethyl acetate (30 mL × 2), the organic phases were combined, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to remove the solvent. Then 5 mL of dichloromethane was added, and 4 mL of formic acid was added. The reaction system was heated to 40 ° C. and acid hydrolyzed at this temperature until the reaction was complete. After the reaction was completed, the reaction was poured into water, extracted with ethyl acetate (30 mL × 2), the organic phases were combined, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product, which was directly used for the next step reaction.

[0072]

[0073] Synthesis of compound A05: Compound II-4 (80 mg, 0.17 mmol), compound VI (70 mg, 0.17 mmol) and PyBOP (135 mg, 0.26 mmol) were dissolved in 5 mL of N,N-dimethylformamide, and DIPEA (120 μL, 0.68 mmol) was added under stirring. The system was reacted at room temperature for 2 h. The reaction was completed under LC-MS monitoring. 15 mL of water was added to the reaction system to precipitate solids, which were filtered and dried to obtain a crude product. The crude product was separated by column chromatography (DCM:MeOH=100:1~30:1) to obtain pure brown-yellow solid powder A05 (55 mg, yield: 38%). 1H NMR(500MHz,DMSO-d6)δ13.46(s,1H),11.11(s,1H),10.90(s,1H),9.29(s,1H),7.76(dd,J=9.3,2.6Hz,1H),7.73(s,1H),7 .65(d,J=3.1Hz,1H),7.58(t,J=7.8Hz,1H),7.14(d,J=8.5Hz,1H),7.04(d,J=7.0Hz,1H),6.94(td,J=9.1,2.6Hz,1H),6.84 (dd,J=8.5,4.5Hz,1H),6.61(t,J=5.8Hz,1H),5.06(dd,J=12.8,5.4Hz,1H),3.63(dd,J=6.5,3.2Hz,4H),3.61–3.39(m,16H ),3.31–3.04(m,1H),2.93–2.85(m,1H),2.61(t,J=3.3Hz,1H),2.56(dt,J=9.7,5.3Hz,4H),2.29(s,2H),2.10–1.99(m,2H). 13 CNMR(126MHz,DMSO-d6)δ173.27,170.80,170.54,169.86,169.39,169.27,167.75,159.5 8,157.72,146.86,136.67,135.18,132.55,127.48,125.68,124.16,121.15,119.48,117. 90,115.76,112.85,111.13,110.49,109.71,106.32,70.29,70.25(3C),70.13,69.34,67. 21,60.22,52.84,52.82,49.03(2C),42.16,33.21,31.45,21.23,9.13.HR-MS(ESI):calcd forC 42 H 48 F8O 10 [M+H] + :843.3471; found:843.3478.

[0074] Example 6

[0075] Synthesis of (Z)-N-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)-9-(4-(2-((5-((5-fluoro-2-oxoindole-3-ylidene)methyl)-4-methyl-1H-pyrrol-3-yl)amino)-2-oxoethyl)piperazin-1-yl)nonanamide (A06)

[0076]

[0077] Synthesis of intermediate II-5: Take compound I-3 (250 mg, 0.92 mmol) and 9-bromononanoic acid (200 mg, 0.84 mmol) and dissolve them in 5 mL of acetonitrile, then add TCFH (356 mg, 1.27 mmol) and NMI (203 μL, 2.5 mmol) and stir the reaction at room temperature for about 2 h. The reaction is completed after LC-MS monitoring. The reaction solution is poured into 20 mL of water, extracted with ethyl acetate (30 mL×2), the organic phases are combined, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and vacuum removed to remove the solvent, and the product is simply purified to obtain a crude compound II-5, which is directly used for the next step reaction.

[0078]

[0079] Synthesis of compound A06: Take compound II-5 (113 mg, 0.23 mmol), compound VI (80 mg, 0.19 mmol), and sodium bicarbonate (50 mg, 0.8 mmol) and dissolve them in 5 mL of N,N-dimethylformamide. Heat the reaction system to 90°C and react under this condition for 4 hours. The reaction is completed after LC-MS monitoring. Cool the reaction system to room temperature, add 20 mL of water, precipitate the solid crude product, and separate by column chromatography (DCM:MeOH=100:1~30:1) to obtain pure brown-yellow solid powder A06 (33 mg, yield: 22%). 1 H NMR (500MHz, DMSO-d6) δ13.45(s,1H),11.13(s,1H),10.91(s,1H),10.67(s,1H),9.24(s,1H),8.28(s,1H),7.93(d,J=10 .1Hz,1H),7.87(d,J=8.2Hz,1H),7.76(dd,J=9.4,2.6Hz,1H),7.73(s,1H),7.65(d,J=3.1Hz,1H),6.94(td,J=9.1,2.6Hz ,1H),6.84(dd,J=8.5,4.5Hz,1H),5.13(dd,J=12.9,5.4Hz,1H),3.20(d,J=5.0Hz,2H)3.06(q,J=7.2Hz,4H),2.94–2.84( m,2H),2.62–2.54(m,4H),2.41(t,J=7.4Hz,4H),2.28(s,3H),2.07–1.99(m,2H),1.65–1.59(m,2H),1.38–1.26(m,10H). 13C NMR(126MHz,DMSO-d6)δ173.22,172.86,170.38,169.85,168.13,167.51,167.24,159.5 7,157.71,145.66,135.18,133.23,127.56,127.48,125.69,125.14,124.27,123.93,116 .69,115.76,113.27,110.49,110.42,106.51,106.31,60.22,49.42,49.06,45.97(4C), 36.98,31.42,29.51,29.13,28.99,25.26,22.53,21.24,14.56,9.08.HR-MS(ESI):calcd for C 42 H 48 FN8O7[M+H] + :795.3625; found:795.3603.

[0080] Example 7

[0081] Synthesis of (Z)-2-(4-(9-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)oxy)nonylpiperazin-1-yl)-N-(5-((5-fluoro-2-oxoindole-3-ylidene)methyl)-4-methyl-1H-pyrrol-3-yl)acetamide (A07)

[0082]

[0083] Synthesis of intermediate II-6: Compound I-1 (200 mg, 0.73 mmol), 1,9-dibromononane (250 mg, 0.88 mmol) and DIPEA (390 μL, 2.2 mmol) were dissolved in 5 mL of N,N-dimethylformamide, the reaction system was heated to 85 °C for 4 h, and the reaction was completed after monitoring by LC-MS. The reaction system was cooled to room temperature and then poured into water. The mixture was extracted with ethyl acetate, the organic phases were combined, washed with water, washed with brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed in vacuo. The product was simply purified to obtain a crude compound II-6, which was directly used for the next step.

[0084]

[0085] Synthesis of compound A07: Take compound II-6 (115 mg, 0.24 mmol), compound VI (84 mg, 0.20 mmol), and sodium bicarbonate (50 mg, 0.8 mmol) and dissolve them in 5 mL of N,N-dimethylformamide. Heat the reaction system to 90°C and react under this condition for 4 hours. The reaction is completed after LC-MS monitoring. Cool the reaction system to room temperature, add 20 mL of water, precipitate the solid crude product, and separate by column chromatography (DCM:MeOH=100:1~30:1) to obtain pure brown-yellow solid powder A07 (36 mg, yield: 23%). 1 H NMR(500MHz,DMSO-d6)δ13.45(s,1H)11.13(s,1H),10.90(s,1H),9.24(s,1H),7.81(t,J=7.9Hz,1H),7.76(d d,J=9.5,2.6Hz,1H),7.73(s,1H),7.65(d,J=3.1Hz,1H),7.51(d,J=8.6Hz,1H),7.44(d,J=7.7Hz,1H),6.98– 6.89(m,1H),6.85(dd,J=8.6,4.4Hz,1H),5.09(dd,J=12.9,5.5Hz,1H),4.20(t,J=6.4Hz,2H),3.19(s,2H),2 .94–2.86(m,2H),2.77–2.49(m,10H),2.28(s,3H),2.10–2.01(m,2H),1.52–1.43(m,4H),1.39–1.25(m,10H). 13 C NMR(126MHz,DMSO-d6)δ173.25,170.42,169.86,167.58,167.31,165.78,159.57,157.71, 156.48,137.50,135.19,133.71,127.56,125.69,125.64,124.27,120.24,119.37,116.66, 115.75,115.59,113.02,110.43,106.51,69.26,67.10,64.01,57.51,49.20(2C),31.43(2C ),29.33,29.19,29.08,28.88,27.00,25.74,22.53,22.48,14.44,9.09.HR-MS(ESI):calcd for C 42 H 49 FN7O7[M+H] + :782.3672; found:782.3670.

[0086] Example 8

[0087] Synthesis of (Z)-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)amino)ethyl)-5-(4-(2-((5-((5-fluoro-2-oxoindole-3-ylidene)methyl)-4-methyl-1H-pyrrol-3-yl)amino)-2-oxoethyl)piperazin-1-yl)-5-oxopentanamide (A08)

[0088]

[0089] Synthesis of intermediate II-7: Compound I-2 (500 mg, 1.81 mmol), N-Boc ethylenediamine (287 μL, 1.81 mmol) and DIPEA (650 μL, 3.62 mmol) were dissolved in 10 mL of DMA, and then the reaction system was heated to 80 ° C. The reaction was allowed to proceed for 2 h under this condition. The reaction was completed by monitoring the system with TLC (dichloromethane: methanol = 10: 1). The system was cooled to room temperature and then poured into water. The mixture was extracted with ethyl acetate (100 mL × 2). The organic phases were combined, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed in vacuo. After simple purification, the obtained product was dissolved in a small amount of ethyl acetate and 15 mL of ethyl acetate was added. HCl-EA, then seal and react at room temperature. After the reaction is completed under TLC monitoring, remove the solvent in vacuo, add 15 mL of ethyl acetate, and stir for 1 hour. Remove the solvent in vacuo again, then add a large amount of ethyl acetate, stir for 0.5 hour, filter, rinse the filter cake with a large amount of ethyl acetate and n-hexane, and dry it with an oil pump to obtain a yellow solid II-7.

[0090]

[0091] Synthesis of intermediate III-3: First, dissolve the linker (1.28 mmol) of alkane diacid and HBTU (180 mg, 0.47 mmol) in 5 mL of N,N-dimethylformamide, add DIPEA (615 μL, 3.44 mmol) and stir at room temperature for 5 min, then add II-7 (150 mg, 0.43 mmol), react at room temperature for 2 h, LC-MS monitored the completion of the reaction, and it can be directly used for the next step after simple treatment.

[0092]

[0093] Synthesis of compound A08: Compound III-3 (103 mg, 0.24 mmol), compound VI (101 mg, 0.24 mmol) and PyBOP (190 mg, 0.36 mmol) were dissolved in 5 mL of N,N-dimethylformamide, and DIPEA (170 μL, 0.96 mmol) was added under stirring. The system was reacted at room temperature for 2 h. The reaction was completed under LC-MS monitoring. 15 mL of water was added to the reaction system to precipitate solids, which were filtered and dried to obtain a crude product. The crude product was separated by column chromatography (DCM:MeOH=100:1~30:1) to obtain pure brown-yellow solid powder A08 (46 mg, yield: 24%). 1 H NMR (500MHz, DMSO-d6) δ13.45(s,1H),11.11(s,1H),10.89(s,1H),9.25(s,1H),8.05(t,J=5.7Hz,1H),7.76(dd,J=9.3,2.6Hz,1H),7. 74(s,1H),7.65(d,J=3.0Hz,1H),7.59(t,J=7.9Hz,1H),7.18(d,J=8.6Hz,1H),7.03(d,J=7.2Hz,1H),6.98–6.90(m,1H),6.84(dd,J=8. 6,4.5Hz,1H),6.74(t,J=6.1Hz,1H),5.06(dd,J=12.8,5.5Hz,1H),3.52(s,2H),3.47(s,2H),3.44–3.36(m,4H),3.26(t,J=6.1Hz,2H), 3.18(d,J=4.6Hz,2H),2.94–2.86(m,1H),2.64–2.60(m,1H),2.59–2.52(m,4H),2.28(s,3H),2.17–1.96(m,4H),1.73(q,J=7.4Hz,2H). 13C NMR (126MHz, DMSO-d6) δ173.27,172.89,170.71,170.56,169.87,169.17,167.76,167.64 ,159.58,157.72,146.81,136.66,135.17,132.67,125.72,125.66,124.28,121.21,119. 52,117.63,115.73,112.82,111.02,110.47,109.70,106.32,61.12,53.42,52.98,49.00 ,45.37,42.00,41.51,38.45,35.07,32.13,31.46,22.64,21.37,9.11.HR-MS(ESI):calcd for C 39 H 41 FN9O8[M+H] + :796.3213; found:796.3207.

[0094] Example 9

[0095] Synthesis of (Z)-N-(2-((2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)amino)ethyl)-6-(4-(2-((5-((5-fluoro-2-oxoisoindol-3-ylidene)methyl)-4-methyl-1H-pyrrol-3-yl)amino)-2-oxoethyl)piperazin-1-yl)-6-oxohexanamide (A09)

[0096]

[0097] Synthesis of intermediate III-4: First, dissolve the linker (1.28 mmol) of alkane diacid and HBTU (180 mg, 0.47 mmol) in 5 mL of N,N-dimethylformamide, add DIPEA (615 μL, 3.44 mmol) and stir at room temperature for 5 min, then add II-7 (150 mg, 0.43 mmol), react at room temperature for 2 h, LC-MS monitors the completion of the reaction, and the next step can be directly used for simple treatment.

[0098]

[0099] Synthesis of compound A09: Compound III-4 (93 mg, 0.21 mmol), compound VI (92 mg, 0.21 mmol) and PyBOP (164 mg, 0.32 mmol) were dissolved in 5 mL of N,N-dimethylformamide, and DIPEA (150 μL, 0.84 mmol) was added under stirring. The system was reacted at room temperature for 2 h. The reaction was completed under LC-MS monitoring. 15 mL of water was added to the reaction system to precipitate solids, which were filtered and dried to obtain a crude product. The crude product was separated by column chromatography (DCM:MeOH=100:1~30:1) to obtain pure brown-yellow solid powder A09 (51 mg, yield: 30%). 1 H NMR (500MHz, DMSO-d6) δ13.45(s,1H),11.11(s,1H),10.89(s,1H),9.24(s,1H),8.05(t,J=5.7Hz,1H),7.76(dd,J=9.4,2.6Hz,1H),7.73(s,1H),7. 64(d,J=3.0Hz,1H),7.59(dd,J=8.6,7.1Hz,1H),7.18(d,J=8.6Hz,1H),7.03(d,J=7.0Hz,1H),6.93(td,J=9.1,2.5Hz,1H), 6.84(dd,J=8.4,4.5Hz, 1H),6.74(t,J=6.2Hz,1H),5.06(dd,J=12.7,5.4Hz,1H),3.54–3.47(m,4 H),3.38(d,J=6.3Hz,2H),3.25(q,J=6.2Hz,2H),3.17(s,2H),2.94–2.86( m,1H),2.63–2.58(m,1H),2.54(d,J=5.0Hz,2H),2.49(s,2H),2.30(d,J= 7.3Hz, 2H), 2.28 (s, 3H), 2.07–1.99 (m, 2H), 1.48 (dq, J = 21.3, 7.7Hz, 4H). 13CNMR(126MHz,DMSO-d6)δ173.27,173.08,170.92,170.56,169.87,169.17,167.76,167.68, 159.57,157.72,146.82,137.47,136.65,135.17,132.67,127.51,125.70,124.28,121.21, 119.51,117.61,115.73,112.82,110.99,110.47,109.70,61.15,53.47,53.01,49.07,49.0 0,45.45,41.95,38.47,35.66,32.49,31.45,25.42,24.90,22.64,9.10.HR-MS(ESI):calcd for C 41 H 45 FN9O8[M+H] + :810.3370; found:810.3364.

[0100] Example 10

[0101] Synthesis of (Z)-N-(2-((2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)amino)ethyl)-7-(4-(2-((5-((5-fluoro-2-oxoindole-3-ylidene)methyl)-4-methyl-1H-pyrrol-3-yl)amino)-2-oxoethyl)piperazin-1-yl)-7-oxoheptylamide (A10)

[0102]

[0103] Synthesis of intermediate III-5: First, dissolve the linker (1.28 mmol) of alkane diacid and HBTU (180 mg, 0.47 mmol) in 5 mL of N,N-dimethylformamide, add DIPEA (615 μL, 3.44 mmol) and stir at room temperature for 5 min, then add II-7 (150 mg, 0.43 mmol), react at room temperature for 2 h, LC-MS monitored the completion of the reaction, and it can be directly used for the next step after simple treatment.

[0104]

[0105] Synthesis of compound A10: Compound III-5 (82 mg, 0.18 mmol), compound VI (76 mg, 0.18 mmol) and PyBOP (140 mg, 0.27 mmol) were dissolved in 5 mL of N,N-dimethylformamide, and DIPEA (127 μL, 0.72 mmol) was added under stirring. The system was reacted at room temperature for 2 h. The reaction was completed under LC-MS monitoring. 15 mL of water was added to the reaction system to precipitate solids, which were filtered and dried to obtain a crude product. The crude product was separated by column chromatography (DCM:MeOH=100:1~30:1) to obtain pure brown-yellow solid powder A10 (47 mg, yield: 32%). 1 H NMR (500MHz, DMSO-d6) δ13.46(s,1H),11.11(s,1H),10.89(s,1H),9.27(s,1H),8.03(t,J=5.7Hz,1H),7.76(dd,J=9.5,2.6Hz,1H),7.73(s,1H ),7.65(d,J=3.0Hz,1H),7.59(t,J=7.9Hz,1H),7.18(d,J=8.5Hz,1H),7.03(d,J=6.1Hz,1H),6.93(t,J=9.2Hz,1H),6.84(dt,J=7.9,3.3Hz,1H ),6.73(t,J=6.1Hz,1H),5.07(dd,J=13.3,5.6Hz,1H),3.51(d,J=12.8Hz,4H),3.46–3.38(m,2H),3.25(d,J=6.6Hz,2H),3.18(s,2H),2.94–2. 85(m,1H),2.64–2.60(m,1H),2.59–2.52(m,4H),2.28(s,3H),2.26(d, J=7.6Hz,2H),2.11–1.98(m,4H),1.56–1.42(m,4H),1.25–1.21(m,2H). 13C NMR(126MHz,DMSO-d6)δ173.27,173.17,171.04,170.55,169.87,169.17,167.79,167.76,15 9.58,157.72,146.83,136.65,135.17,132.67,127.58,125.70,124.26,121.20,119.50,117 .63,115.75,113.02,111.00,110.48,109.70,106.51,61.09,55.38,53.44,52.99,49.07,49 .00,41.95,38.44,35.79,32.58,31.45,28.89,25.56,25.06,22.64,9.11.HR-MS(ESI):calcd forC 42 H 47 FN9O8[M+H] + :824.3527; found:824.3522.

[0106] Embodiment 11

[0107] Synthesis of (Z)-N-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)amino)ethyl)-8-(4-(2-((5-((5-fluoro-2-oxoindole-3-ylidene)methyl)-4-methyl-1H-pyrrol-3-yl)amino)-2-oxoethyl)piperazin-1-yl)-8-oxooctylamide (A11)

[0108]

[0109] Synthesis of intermediate III-6: First, dissolve the linker (1.28 mmol) of alkane diacid and HBTU (180 mg, 0.47 mmol) in 5 mL of N,N-dimethylformamide, add DIPEA (615 μL, 3.44 mmol) and stir at room temperature for 5 min, then add II-7 (150 mg, 0.43 mmol), react at room temperature for 2 h, and monitor the completion of the reaction by LC-MS. After simple treatment, it can be directly used for the next step.

[0110]

[0111] Synthesis of compound A11: Compound III-6 (138 mg, 0.29 mmol), compound VI (121 mg, 0.29 mmol) and PyBOP (225 mg, 0.44 mmol) were dissolved in 5 mL of N,N-dimethylformamide, and DIPEA (205 μL, 1.16 mmol) was added under stirring. The system was reacted at room temperature for 2 h. The reaction was completed under LC-MS monitoring. 15 mL of water was added to the reaction system to precipitate solids, which were filtered and dried to obtain a crude product. The crude product was separated by column chromatography (DCM:MeOH=100:1~30:1) to obtain pure brown-yellow solid powder A11 (64 mg, yield: 26%). 1 H NMR (500MHz, DMSO-d6) δ13.45(s,1H),11.11(s,1H),10.89(s,1H),9.24(s,1H),8.02(t,J=5.7Hz,1H),7.76(dd,J=9.4,2.6Hz,1H),7.73(s,1H),7. 64(d,J=3.0Hz,1H),7.59(t,J=8.7Hz,1H),7.18(d,J=8.7Hz,1H),7.03(d, J=7.0Hz,1H),6.96–6.90(m,1H),6.84(dd,J=8.4,4.5Hz,1H),6.72(t,J=6 .2Hz,1H),5.06(dd,J=12.7,5.5Hz,1H),3.51(dt,J=10.2,4.6Hz,4H),3.3 8(d,J=6.4Hz,2H),3.25(q,J=6.1Hz,2H),3.18(s,2H),2.94–2.86(m,1H), 2.65–2.58(m,1H),2.54(d,J=5.0Hz,4H),2.49(s,2H),2.28(s,3H),2.26( d,J=7.5Hz,2H),2.07–2.03(m,2H),1.56–1.38(m,4H),1.31–1.20(m,4H). 13C NMR(126MHz,DMSO-d6)δ173.28,173.20,171.06,170.56,169.87,169.17,167.76,167.69,159 .57,157.71,146.83,136.65,135.16,132.67,127.57,125.71,124.29,121.22,119.52,117.62 ,115.72,112.81,111.01,110.39,109.69,106.51,61.15,53.49,53.01,48.99,45.47,41.92,4 1.51,38.44,35.84,32.67,31.44,29.03,28.99,25.58,25.16,22.64,9.10.HR-MS(ESI):calcd for C 43 H 49 FN9O8[M+H] + :838.3682; found:838.3654.

[0112] Example 12

[0113] Synthesis of (Z)-N-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)amino)ethyl)-9-(4-(2-((5-((5-fluoro-2-oxoisoindol-3-ylidene)methyl)-4-methyl-1H-pyrrol-3-yl)amino)-2-oxoethyl)piperazin-1-yl)-9-oxononamide (A12)

[0114]

[0115] Synthesis of intermediate III-7: First, dissolve the linker (1.28 mmol) of alkane diacid and HBTU (180 mg, 0.47 mmol) in 5 mL of N,N-dimethylformamide, add DIPEA (615 μL, 3.44 mmol) and stir at room temperature for 5 min, then add II-7 (150 mg, 0.43 mmol), react at room temperature for 2 h, and the reaction is completed under LC-MS monitoring. After simple treatment, it can be directly used for the next step.

[0116]

[0117] Synthesis of compound A12: Compound III-7 (118 mg, 0.24 mmol), compound VI (100 mg, 0.24 mmol) and PyBOP (135 mg, 0.26 mmol) were dissolved in 5 mL of N,N-dimethylformamide, and DIPEA (120 μL, 0.68 mmol) was added under stirring. The system was reacted at room temperature for 2 h. The reaction was completed under LC-MS monitoring. 15 mL of water was added to the reaction system to precipitate the solid, which was filtered and dried to obtain the crude product. The crude product was separated by column chromatography (DCM:MeOH=100:1~30:1) to obtain pure brown-yellow solid powder A12 (45 mg, yield: 22%). 1 H NMR (500MHz, DMSO-d6) δ13.50(s,1H),11.11(s,1H),10.96(s,1H),10.34(s,1H),8.08(t,J=5.7Hz,1H),7.77(dd,J=9.3,2.6Hz,1H),7. 73(s,1H),7.71(d,J=3.1Hz,1H),7.59(t,J=7.8Hz,1H),7.18(d,J=8.5Hz,1H),7.03(d,J=6.9Hz,1H),6.95(td,J=9.0,2.5Hz,1H),6.86 (dd,J=8.4,4.5Hz,1H),6.72(t,J=6.2Hz,1H),5.06(dd,J=12.8,5.4Hz,1H),3.60–3.51(m,4H),3.34–3.20(m,4H),3.11(s,2H),2.93–2 .85(m,1H),2.61(s,1H),2.58–2.51(m,4H),2.50(s,2H),2.34(s,3H),2.10–1.97(m,4H),1.56–1.40(m,4H),1.23(q,J=7.1,6.6Hz,6H). 13C NMR (126MHz, DMSO-d6) δ173.28,173.24,171.42,170.54,169.83,169.14,167.77,162.35,159.5 7,157.71,146.83,136.67,135.30,132.64,125.67,125.63,123.70,120.90,119.29,117.68,11 6.21,113.00,111.01,110.61,109.65,106.41,56.78,52.15,51.96,49.06,48.98,42.10,41.96 ,38.43,35.82,32.36,31.47,29.10,29.06,29.01,25.63,24.96,22.66,9.49.HR-MS(ESI):calcd forC 44 H 51 FN9O8[M+H] + :852.3840; found:852.3819.

[0118] Example 13

[0119] Synthesis of ((Z)-N-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)amino)ethyl)-10-(4-(2-((5-((5-fluoro-2-oxoindole-3-ylidene)methyl)-4-methyl-1H-pyrrol-3-yl)amino)-2-oxoethyl)piperazin-1-yl)-10-oxodecanoic acid amide (A13)

[0120]

[0121] Synthesis of intermediate III-8: First, dissolve the linker (1.28 mmol) of alkane diacid and HBTU (180 mg, 0.47 mmol) in 5 mL of N,N-dimethylformamide, add DIPEA (615 μL, 3.44 mmol) and stir at room temperature for 5 min, then add II-7 (150 mg, 0.43 mmol), react at room temperature for 2 h, and monitor the completion of the reaction by LC-MS. After simple treatment, it can be directly used for the next step.

[0122]

[0123] Synthesis of compound A13: Compound III-8 (116 mg, 0.23 mmol), compound VI (96 mg, 0.23 mmol) and PyBOP (182 mg, 0.35 mmol) were dissolved in 5 mL of N,N-dimethylformamide, and DIPEA (163 μL, 0.92 mmol) was added under stirring. The system was reacted at room temperature for 2 h. The reaction was completed under LC-MS monitoring. 15 mL of water was added to the reaction system to precipitate solids, which were filtered and dried to obtain a crude product. The crude product was separated by column chromatography (DCM:MeOH=100:1~30:1) to obtain pure brown-yellow solid powder A13 (54 mg, yield: 27%). 1 H NMR (500MHz, DMSO-d6) δ13.45(s,1H),11.11(s,1H),10.89(s,1H),9.25(s,1H),8.02(t,J=5.6Hz,1H),7.76(dd,J=9.3,2.6Hz,1H),7.73(s,1H), 7.64(d,J=3.1Hz,1H),7.58(t,J=8.6Hz,1H),,7.17(d,J=8.6Hz,1H),7.0 3(d,J=7.0Hz,1H),6.93(td,J=9.5,9.0,2.6Hz,1H),6.84(dd,J=8.5,4.5 Hz,1H),6.72(t,J=6.1Hz,1H),5.06(dd,J=12.7,5.4Hz,1H),3.51(q,J=6.0,5.0Hz,4H),3.37(d,J=6.3Hz,2H),3.24(q,J=6.1Hz,2H),3.18(s,2H) ,2.94–2.86(m,1H),2.61(d,J=3.3Hz,1H),2.58–2.51(m,4H),2.50–2.45 (m,2H),2.28(s,3H),2.06–1.98(m,4H),1.50–1.44(m,4H),1.22(s,8H). 13C NMR(126MHz,DMSO-d6)δ173.26,173.21,171.07,170.80,170.53,169.87,169.16,167.76,159.58 ,157.72,146.83,136.63,135.17,132.66,127.58,125.66,124.27,121.19,119.50,117.61,115.7 4,112.81,110.99,110.47,109.68,106.51,61.15,60.22,53.48,53.02,49.07,48.99,41.94,38.4 5,35.87,32.72,31.45,29.24,29.21,29.09,25.68,25.29,22.65,21.23,9.10.HR-MS(ESI):calcd for C 45 H 53 FN9O8[M+H] + :866.3996; found:866.3989.

[0124] Embodiment 14

[0125] Synthesis of (Z)-2-(4-(6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)amino)hexyl)piperazin-1-yl)-N-(5-((5-fluoro-2-oxoindole-3-ylidene)methyl)-4-methyl-1H-pyrrol-3-yl)acetamide (A14)

[0126]

[0127] Synthesis of intermediate II-8: Take I-2 (500 mg, 1.81 mmol), aminoalkane acid linker (1.81 mmol) and DIPEA (650 μL, 3.62 mmol) and dissolve them in 10 mL of DMA, then heat the reaction system to 80°C and react for 2 hours under this condition. The reaction is completed by monitoring by TLC, and the system is cooled to room temperature and poured into water. Extract with ethyl acetate (100 mL×2), combine the organic phases, wash with water, wash with saturated brine, dry with anhydrous sodium sulfate, filter, remove the solvent in vacuo, and directly use it for the next reaction after simple treatment.

[0128]

[0129] Synthesis of compound A14: Compound II-8 (110 mg, 0.26 mmol), compound VI (110 mg, 0.26 mmol) and PyBOP (203 mg, 0.39 mmol) were dissolved in 5 mL of N,N-dimethylformamide, and DIPEA (190 μL, 1.04 mmol) was added under stirring. The system was reacted at room temperature for 2 h. The reaction was completed under LC-MS monitoring. 15 mL of water was added to the reaction system to precipitate the solid, which was filtered and dried to obtain the crude product. The crude product was separated by column chromatography (DCM:MeOH=100:1~30:1) to obtain pure brown-yellow solid powder A14 (75 mg, yield: 38%). 1 H NMR (500MHz, DMSO-d6) δ13.46(s,1H),11.11(s,1H),10.89(s,1H),9.27(s,1H),7.76(dd,J=9.4,2.6Hz,1H),7.74(s,1H),7.65(d,J=3.0 Hz,1H),7.59(t,J=8.0Hz,1H),7.10(d,J=8.6Hz,1H),7.02(d,J=7.1Hz,1H),6.94(t,J=9.2Hz,1H),6.84(dd,J=8.7,4.5Hz,1H),6.54(t, J=6.0Hz,1H),5.06(dd,J=12.8,5.5Hz,1H),3.52(s,4H),3.30(d,J=7.6Hz,2H),3.20(s,2H),2.93–2.84(m,1H),2.61(s,1H),2.59–2.52 (m,4H),2.33(t,J=7.6Hz,2H),2.28(s,3H),2.10–1.99(m,2H),1.60(t,J=7.5Hz,2H),1.54(q,J=8.1,7.6Hz,2H),1.37(p,J=7.4Hz,2H). 13C NMR (126MHz, DMSO-d6) δ173.28,171.00,170.57,169.87,169.41,168.43,167.77,159. 58,157.72,146.89,136.75,135.18,132.66,125.70,125.66,124.26,121.19,119.50, 117.65,115.75,113.02,110.84,110.48,109.46,106.31,61.06,55.38,53.45,53.00, 49.01(2C),42.24,32.61,31.45,29.03,26.57,25.02,22.63,9.11.HR-MS(ESI):calcd for C 39 H 42 FN8O7[M+H] + :753.3155; found:753.3144.

[0130] Embodiment 15

[0131] Synthesis of (Z)-2-(4-(7-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)amino)heptanoyl)piperazin-1-yl)-N-(5-((5-fluoro-2-oxoindole-3-ylidene)methyl)-4-methyl-1H-pyrrol-3-yl)acetamide (A15)

[0132]

[0133] Synthesis of intermediate II-9: Take I-2 (500 mg, 1.81 mmol), aminoalkane acid linker (1.81 mmol) and DIPEA (650 μL, 3.62 mmol) and dissolve them in 10 mL of DMA, then heat the reaction system to 80°C and react for 2 hours under this condition. The reaction is completed by monitoring by TLC, and the system is cooled to room temperature and poured into water. Extract with ethyl acetate (100 mL×2), combine the organic phases, wash with water, wash with saturated brine, dry with anhydrous sodium sulfate, filter, remove the solvent in vacuo, and directly use it for the next reaction after simple treatment.

[0134]

[0135] Synthesis of compound A15: Compound II-9 (85 mg, 0.21 mmol), compound VI (90 mg, 0.21 mmol) and PyBOP (165 mg, 0.32 mmol) were dissolved in 5 mL of N,N-dimethylformamide, and DIPEA (150 μL, 0.84 mmol) was added under stirring. The system was reacted at room temperature for 2 h. The reaction was completed under LC-MS monitoring. 15 mL of water was added to the reaction system to precipitate the solid, which was filtered and dried to obtain the crude product. The crude product was separated by column chromatography (DCM:MeOH=100:1~30:1) to obtain pure brown-yellow solid powder A15 (45 mg, yield: 28%). 1 H NMR (500MHz, DMSO-d6) δ13.46(s,1H),11.11(s,1H),10.89(s,1H),9.27(s,1H),7.76(dd,J=9.3,2.6Hz,1H),7.73(s,1H),7.64(d,J=3.0Hz ,1H),7.58(t,J=7.8Hz,1H),7.09(d,J=8.6Hz,1H),7.02(d,J=7.1Hz,1H),6.93(td,J=9.1,2.6Hz,1H),6.84(dd,J=8.5,4.5Hz,1H),6.54(t, J=5.9Hz,1H),5.06(dd,J=12.8,5.4Hz,1H),3.57–3.44(m,4H),3.29(t,J=6.8Hz,2H),3.20(s,2H),2.92–2.85(m,1H),2.61(t,J=3.3Hz,1H) ,2.59–2.51(m,4H),2.31(t,J=7.5Hz,2H),2.28(s,3H),2.12–1.97(m,2H),1.57(q,J=7.2Hz,2H),1.50(q,J=7.3Hz,2H),1.40–1.30(m,4H). 13C NMR (126MHz, DMSO-d6) δ173.28,171.05,170.56,169.96,169.86,169.41,167.76,159.5 8,157.71,146.89,136.74,135.17,132.66,127.57,125.70,124.24,121.18,119.50,11 7.64,115.75,112.83,110.84,110.47,109.48,106.31,61.03,60.96,53.43,53.42,52. 98,49.01(2C),32.61,31.45,29.06,28.97,26.63,25.22,22.63,9.11.HRMS(ESI):calcd for C 40 H 44 FN8O7[M+H] + :767.3311; found:767.3304.

[0136] Example 16

[0137] Synthesis of (Z)-2-(4-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)amino)octyl)piperazin-1-yl)-N-(5-((5-fluoro-2-oxoindole-3-ylidene)methyl)-4-methyl-1H-pyrrol-3-yl)acetamide (A16)

[0138]

[0139] Synthesis of intermediate II-10: Take I-2 (500 mg, 1.81 mmol), aminoalkane acid linker (1.81 mmol) and DIPEA (650 μL, 3.62 mmol) and dissolve them in 10 mL of DMA, then heat the reaction system to 80°C and react for 2 hours under this condition. The reaction is completed by monitoring by TLC, and the system is cooled to room temperature and poured into water. Extract with ethyl acetate (100 mL×2), combine the organic phases, wash with water, wash with saturated brine, dry with anhydrous sodium sulfate, filter, remove the solvent in vacuo, and directly use it for the next reaction after simple treatment.

[0140]

[0141] Synthesis of compound A16: Compound II-10 (80 mg, 0.19 mmol), compound VI (82 mg, 0.19 mmol) and PyBOP (150 mg, 0.29 mmol) were dissolved in 5 mL of N,N-dimethylformamide, and DIPEA (140 μL, 0.76 mmol) was added under stirring. The system was reacted at room temperature for 2 h. The reaction was completed under LC-MS monitoring. 15 mL of water was added to the reaction system to precipitate the solid, which was filtered and dried to obtain the crude product. The crude product was separated by column chromatography (DCM:MeOH=100:1~30:1) to obtain pure brown-yellow solid powder A16 (52 mg, yield: 35%). 1 H NMR (500MHz, DMSO-d6) δ13.46(s,1H),11.11(s,1H),10.89(s,1H),9.25(s,1H),7.76(dd,J=9.3,2.6Hz,1H),7.73(s,1H),7.64(d,J=3.1H z,1H),7.59(t,J=8.0Hz,1H),7.09(d,J=8.6Hz,1H),7.02(d,J=7.0Hz,1H),6.93(td,J=9.1,2.6Hz,1H),6.84(dd,J=8.5,4.5Hz,1H),6.53( t,J=5.9Hz,1H),5.06(dd,J=12.8,5.4Hz,1H),3.51(s,4H),3.29(q,J=6.7Hz,2H),3.18(s,2H),2.94–2.86(m,1H),2.65–2.58(m,1H),2.5 9–2.51(m,4H),2.31(d,J=7.4Hz,2H),2.28(s,3H),2.09–1.96(m,2H),1.61–1.54(m,2H),1.52–1.46(m,2H),1.31(q,J=13.2,10.8Hz,6H). 13C NMR(126MHz,DMSO-d6)δ173.27,171.07,170.56,169.99,169.87,169.42,167.76,159.58, 157.71,146.89,136.73,135.17,132.65,127.58,125.70,124.27,121.19,119.50,117.64 ,115.73,113.01,110.83,110.47,109.47,106.51,61.13,60.22,55.38,53.47,53.02,49. 01(2C),32.68,31.45,29.21,29.13,29.06,26.72,25.24,22.62,9.10.HR-MS(ESI):calcd for C 41 H 46 FN8O7[M+H] + :781.3468; found:781.3465. Example 17

[0142] Synthesis of (Z)-2-(4-(11-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)amino)undecyl)piperazin-1-yl)-N-(5-((5-fluoro-2-oxoindole-3-ylidene)methyl)-4-methyl-1H-pyrrol-3-yl)acetamide (A17)

[0143]

[0144] Synthesis of intermediate II-11: Take I-2 (500 mg, 1.81 mmol), aminoalkane acid linker (1.81 mmol) and DIPEA (650 μL, 3.62 mmol) and dissolve them in 10 mL of DMA, then heat the reaction system to 80°C and react for 2 hours under this condition. The reaction is completed by monitoring by TLC, and the system is cooled to room temperature and poured into water. Extract with ethyl acetate (100 mL×2), combine the organic phases, wash with water, wash with saturated brine, dry with anhydrous sodium sulfate, filter, remove the solvent in vacuo, and directly use it for the next reaction after simple treatment.

[0145]

[0146] Synthesis of compound A17: Compound II-11 (71 mg, 0.16 mmol), compound VI (66 mg, 0.16 mmol) and PyBOP (125 mg, 0.23 mmol) were dissolved in 5 mL of N,N-dimethylformamide, and DIPEA (110 μL, 0.62 mmol) was added under stirring. The system was reacted at room temperature for 2 h. The reaction was completed under LC-MS monitoring. 15 mL of water was added to the reaction system to precipitate the solid, which was filtered and dried to obtain the crude product. The crude product was separated by column chromatography (DCM:MeOH=100:1~30:1) to obtain pure brown-yellow solid powder A17 (55 mg, yield: 41%). 1 H NMR (500MHz, DMSO-d6) δ13.46(s,1H),11.11(s,1H),10.89(s,1H),9.27(s,1H),7.76(dd,J=9.4,2.6Hz,1H),7.73(s,1H),7.65(d,J=3. 1Hz,1H),7.58(t,J=7.8Hz,1H),7.08(d,J=8.6Hz,1H),7.02(d,J=7.1Hz,1H),6.93(t,J=9.2Hz,1H),6.84(dd,J=8.6,4.5Hz,1H),6.52( t,J=5.9Hz,1H),5.06(dd,J=12.8,5.4Hz,1H),3.52(s,4H),3.29(t,J=6.8Hz,2H),3.20(s,2H),2.94–2.86(m,1H),2.61(s,1H),2.59–2 .52(m,4H),2.29(d,J=10.0Hz,5H),2.10–1.99(m,2H),1.57(t,J=7.2Hz,2H),1.50–1.44(m,2H),1.35–1.31(m,2H),1.30–1.20(m,10H). 13C NMR (126MHz, DMSO-d6) δ173.27,171.09,170.80,170.55,169.87,169.41,167.76,159.58,157. 72,146.89,136.72,135.18,132.65,125.70,125.67,124.25,121.16,119.48,117.63,115.76, 110.83,110.47,110.41,109.46,106.51,60.22(2C),53.43,52.99,49.01(2C),42.30,32.70,3 1.45,29.44,29.39,29.34,29.26,29.22,29.13,26.79,25.30,21.23,9.11.HR-MS(ESI):calcd for C 44 H 52 FN8O7[M+H] + :823.3938; found:823.3909.

[0147] Embodiment 18

[0148] Synthesis of (Z)-2-(4-(12-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)amino)dodecyl)piperazin-1-yl)-N-(5-((5-fluoro-2-oxoindole-3-ylidene)methyl)-4-methyl-1H-pyrrol-3-yl)acetamide (A18)

[0149]

[0150] Synthesis of intermediate II-12: Take I-2 (500 mg, 1.81 mmol), aminoalkane acid linker (1.81 mmol) and DIPEA (650 μL, 3.62 mmol) and dissolve them in 10 mL of DMA, then heat the reaction system to 80°C and react for 2 hours under this condition. The reaction is completed by monitoring by TLC, and the system is cooled to room temperature and poured into water. Extract with ethyl acetate (100 mL×2), combine the organic phases, wash with water, wash with saturated brine, dry with anhydrous sodium sulfate, filter, remove the solvent in vacuo, and directly use it for the next reaction after simple treatment.

[0151]

[0152] Synthesis of compound A18: Compound II-12 (150 mg, 0.32 mmol), compound VI (133 mg, 0.32 mmol) and PyBOP (250 mg, 0.48 mmol) were dissolved in 5 mL of N,N-dimethylformamide, and DIPEA (230 μL, 1.28 mmol) was added under stirring. The system was reacted at room temperature for 2 h. The reaction was completed under LC-MS monitoring. 15 mL of water was added to the reaction system to precipitate the solid, which was filtered and dried to obtain the crude product. The crude product was separated by column chromatography (DCM:MeOH=100:1~30:1) to obtain pure brown-yellow solid powder A18 (88 mg, yield: 33%). 1 H NMR (500MHz, DMSO-d6) δ13.45(s,1H),11.10(s,1H),10.88(s,1H),9.24(s,1H),7.76(dd,J=9.4,2.6Hz,1H),7.73(s,1H),7.64(d,J=3.0Hz,1H ),7.58(t,J=8.6Hz,1H),7.09(d,J=8.6Hz,1H),7.02(d,J=7.0Hz,1H),6.93(td,J=9.1,2.6Hz,1H),6.84(dd,J=8.5,4.5Hz,1H),6.52(t,J=5.9 Hz,1H),5.05(dd,J=12.7,5.5Hz,1H),3.51(s,4H),3.31–3.27(m,2H),3.18(s,2H),2.92–2.85(m,1H),2.61(d,J=3.3Hz,1H),2.59–2.52(m,4H) ),2.30(d,J=7.2Hz,2H),2.28(s,3H),2.10–1.99(m,2H),1.57(t,J=7.2Hz,2H),1.48(t,J=7.4Hz,2H),1.36–1.32(m,4H),1.28–1.22(m,10H). 13C NMR(126MHz,DMSO-d6)δ173.26,171.08,170.79,170.55,169.87,169.41,167.76,159.58,157.72 ,146.89,136.72,135.18,132.65,127.58,125.69,124.26,121.16,119.48,117.62,115.75,113. 01,112.82,110.82,109.46,106.51,61.08,60.22,53.45,53.00,49.01(2C),42.30,32.71,31.45 ,29.46,29.43,29.36,29.26,29.23,29.14,26.78,25.31,22.63,21.23,9.10.HR-MS(ESI):calcd for C 45 H 54 FN8O7[M+H] + :837.4093; found:837.4074.

[0153] Embodiment 19

[0154] Synthesis of (Z)-2-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)piperidin-4-yl)methyl)piperazin-1-yl)-N-(5-((5-fluoro-2-oxoindole-3-ylidene)methyl)-4-methyl-1H-pyrrol-3-yl)acetamide (A19)

[0155]

[0156] Synthesis of intermediate II-13: Compound I-2 (500 mg, 1.8 mmol), piperidin-4-ylmethanol (255 μL, 2.2 mmol) and DIPEA (1.6 mL, 9 mmol) were dissolved in 5 mL of dimethyl sulfoxide, the reaction system was heated to 80 ° C, and the reaction was carried out under this condition for 4 hours. The reaction was completed under LC-MS monitoring, the reaction system was cooled to room temperature, and then poured into water, extracted with ethyl acetate, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed in vacuo. After simple treatment, it can be directly used for the next reaction.

[0157]

[0158] Synthesis of intermediate III-9: Dissolve II-13 (540 mg, 1.45 mmol) in 15 mL of dichloromethane, then slowly add Dess-Martin reagent (1.24 g, 2.9 mmol), react at room temperature for 2 h, and after the reaction is completed by LC-MS monitoring, add excess saturated aqueous sodium thiosulfate and saturated aqueous sodium bicarbonate to the reaction system, stir for another 10 min, extract with dichloromethane, wash with water, wash with saturated brine, dry with anhydrous sodium sulfate, filter, remove the solvent in vacuo, and directly use it for the next reaction after simple treatment.

[0159]

[0160] Synthesis of intermediate VII: Compound VI (0.1 g, 0.24 mmol) was dissolved in 10 mL of water, and saturated aqueous sodium bicarbonate solution was added under stirring until the system became alkaline. Then 30 mL of ethyl acetate was added to the system and stirred for about 0.5 h. The layers were separated using a separatory funnel and extracted with ethyl acetate (30 mL × 2). The organic phases were combined, washed with water and brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound VII (0.06 g, yield: 65%). 1 H NMR (500MHz, DMSO-d6) δ13.45(s,1H),10.90(s,1H),9.24(s,1H),7.77(dd,J=9.3,2.6Hz,1H),7.74(s,1H),7.65(d,J=3.2Hz,1H),6.94 (td,J=9.1,2.6Hz,1H),6.84(dd,J=8.4,4.5Hz,1H),3.11(s,2H),2.82(t,J=4.7Hz,4H),2.50–2.41(m,4H),2.28(s,3H),1.24(brs,1H).

[0161]

[0162] Synthesis of compound A19: Compound III-9 (60 mg, 0.16 mmol), compound VII (75 mg, 0.195 mmol) and acetic acid (1 mg, 0.016 mmol) were dissolved in 5 mL of N,N-dimethylformamide, stirred at room temperature for 10 min, and then sodium triacetoxyborohydride (42 mg, 0.195 mmol) was slowly added, and the mixture was reacted at room temperature. The reaction was completed under LC-MS monitoring. The reaction solution was poured into water, extracted with ethyl acetate, and separated by column chromatography (DCM:MeOH=100:1~30:1) to obtain pure brown-yellow solid powder A19 (44 mg, yield: 37%). 1H NMR (500MHz, DMSO-d6) δ13.45(s,1H),11.09(s,1H),10.88(s,1H),9.19(s,1H),7.76(dd,J=9.4,2.6Hz,1H),7.73(s, 1H),7.69–7.65(m,2H),7.32(dd,J=7.8,5.4Hz,2H),6.93(td,J=9.1,2.6Hz,1H),6.84(dd,J=8.4,4.5Hz,1H),5.09(d d,J=12.7,5.5Hz,1H),3.69(d,J=11.6Hz,2H),3.14(s,2H),2.92–2.83(m,3H),2.67–2.53(m,6H),2.49–2.37(m,3H), 2.28(s,3H),2.23(s,2H),2.03(ddd,J=12.3,6.8,4.1Hz,1H),1.82(d,J=12.5Hz,2H),1.73(s,1H),1.41–1.22(m,3H). 13 C NMR(126MHz,DMSO-d6)δ173.25,170.48,169.87,167.56,166.74,159.59,157.72,150 .64,136.17,135.19,134.16,127.50,125.62,124.35,120.60,119.13,116.74,115.7 9,114.78,113.02,112.83,110.48,106.51,106.30,64.35,61.43,53.79,53.45,51.4 8,51.46,51.43,49.25(2C),32.70,31.44,30.94(2C),22.56,8.98.HR-MS(ESI):calcd for C 39 H 42 FN8O6[M+H] + :737.3206; found:737.3196. Example 20

[0163] Synthesis of (Z)-2-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl)piperidin-4-yl)methyl)piperazin-1-yl)-N-(5-((5-fluoro-2-oxoindole-3-ylidene)methyl)-4-methyl-1H-pyrrol-3-yl)acetamide (A20)

[0164]

[0165] Synthesis of intermediate II-14: Compound I-4 (500 mg, 1.8 mmol), piperidin-4-ylmethanol (255 μL, 2.2 mmol) and DIPEA (1.6 mL, 9 mmol) were dissolved in 5 mL of dimethyl sulfoxide, the reaction system was heated to 80 ° C, and the reaction was carried out under this condition for 4 hours. The reaction was completed by LC-MS monitoring, the reaction system was cooled to room temperature, and then poured into water, extracted with ethyl acetate, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, the solvent was removed in vacuo, and separated by column chromatography to obtain compound II-14 (340 mg, yield: 81%). 1 H NMR (500MHz, CDCl3) δ8.46(s,1H),7.68(d,J=8.5Hz,1H),7.29(d,J=2.4Hz,1H),7.06(dd ,J=8.6,2.4Hz,1H),4.95(dd,J=12.1,5.4Hz,1H),3.99(dt,J=13.3,3.3Hz,2H),3.55(d, J=6.3Hz,2H),3.00(ddd,J=15.3,12.0,2.7Hz,2H),2.91–2.77(m,3H),2.14(ddd,J=10.4 ,4.8,2.6Hz,1H),1.93–1.77(m,3H),1.37(td,J=12.4,4.0Hz,2H),1.27(t,J=7.1Hz,1H).

[0166]

[0167] Synthesis of intermediate III-10: II-14 (210 mg, 0.57 mmol) was dissolved in 10 mL of dichloromethane, and then DMP (480 mg, 1.13 mmol) was slowly added. The reaction was allowed to react for 2 h at room temperature. After the reaction was completed by LC-MS monitoring, an excess of saturated aqueous sodium thiosulfate and saturated aqueous sodium bicarbonate were added to the reaction system, and the mixture was stirred for 10 min. The mixture was extracted with dichloromethane, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed in vacuo. The mixture was then separated by column chromatography to obtain the product III-10 (190 mg, yield: 90%). 1H NMR (500MHz, CDCl3) δ9.74(d,J=0.9Hz,1H),8.09(s,1H),7.72(d,J=8.5Hz,1H),7.31(d,J=2.3Hz,1H),7.09(dd,J=8.6,2.4Hz,1H),4.96(dd,J =12.4,5.4Hz,1H),3.88(dt,J=13.4,4.3Hz,2H),3.20(ddd,J=13.4,10. 5,3.1Hz,2H),2.95–2.72(m,4H),2.18–2.07(m,3H),1.85–1.77(m,2H).

[0168]

[0169] Synthesis of compound A20: Compound III-10 (60 mg, 0.16 mmol), compound VII (75 mg, 0.195 mmol) and acetic acid (1 mg, 0.016 mmol) were dissolved in 5 mL of N,N-dimethylformamide, stirred at room temperature for 10 min, then sodium triacetoxyborohydride (42 mg, 0.195 mmol) was slowly added, and the mixture was reacted at room temperature. The reaction was completed after LC-MS monitoring. The reaction solution was poured into water, extracted with ethyl acetate, and separated by column chromatography (DCM:MeOH=100:1~30:1) to obtain pure brown-yellow solid powder A20 (36 mg, yield: 30%). 1 H NMR(500MHz,DMSO-d6)δ13.45(s,1H),11.08(s,1H),10.88(s,1H),9.18(s,1H),7.76(dd,J=9.3,2.6Hz,1H),7.73(s,1H) ,7.68–7.63(m,2H),7.31(d,J=2.3Hz,1H),7.23(dd,J=8.8,2.3Hz,1H),6.93(td,J=9.1,2.6Hz,1H),6.84(dd,J=8.5,4.5H z,1H),5.07(dd,J=12.8,5.4Hz,1H),4.04(d,J=12.9Hz,2H),3.14(s,2H),2.96(t,J=12.5Hz,2H),2.91–2.84(m,1H),2.67 –2.54(m,6H),2.45(s,3H),2.28(s,3H),2.21–2.15(m,2H),2.04–2.00(m,1H),1.80(d,J=13.2Hz,3H),1.27–1.12(m,3H). 13CNMR(126MHz,DMSO-d6)δ173.24,170.55,169.87,168.10,167.69,167.42,157.72,15 5.48,135.19,134.51,127.58,125.66,125.45,124.31,120.61,119.14,118.04,117.8 2,115.79,113.02,110.48,108.18,106.51,106.30,64.13,61.44,53.75,53.43,49.2 2(2C),47.72,32.99,31.46,30.06(2C),22.67,22.52,14.42,8.97.HR-MS(ESI):calcd forC 39 H 42 FN8O6[M+H] + :737.3206; found:737.3193.

[0170] Embodiment 21

[0171] Synthesis of (Z)-2-(4-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl)piperidin-4-yl)piperazin-1-yl)-N-(5-((5-fluoro-2-oxoindole-3-ylidene)methyl)-4-methyl-1H-pyrrol-3-yl)acetamide (A21)

[0172]

[0173] Synthesis of intermediate VIII-1: Compound VII (100 mg, 0.26 mmol), tert-butyl4-oxopiperidine-1-carboxylate (80 mg, 0.40 mmol) and AcOH (1 μL, 0.03 mmol) were dissolved in 4 mL of dimethyl sulfoxide, and then stirred at 25°C for 10 min. Sodium triacetoxyborohydride (80 mg, 0.40 mmol) was added, and the mixture was reacted at 25°C for 2 h. The reaction was completed after LC-MS monitoring. 20 mL of water was poured into the reaction system, and the mixture was extracted with ethyl acetate (30 mL×2). The organic phases were combined, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and vacuum removed to remove the solvent, and the crude product was slurried with (ethyl acetate: n-hexane = 5:1) to obtain product VIII-1-Boc (126 mg, yield: 88%). 1H NMR (500MHz, DMSO-d6) δ13.45(s,1H),10.89(s,1H),9.23(d,J=37.4Hz,1H),7.77(dd,J=9.3,2.6Hz,1H) ,7.73(s,1H),7.62(dd,J=24.5,3.1Hz,1H),6.94(td,J=9.1,2.5Hz,1H),6.84(dd,J=8.4,4.5Hz,1H),4.0 2–3.88 (m, 2H), 3.61 (t, J = 6.3 Hz, 2H), 3.12 (s, 2H), 2.92 (d, J = 12.1 Hz, 1H), 2.77–2.54 (m, 8H), 2.27 (s, 3H), 1.75 (d, J = 12.4 Hz, 2H), 1.40 (s, 9H), 1.31–1.23 (m, 2H). VIII-1 was obtained by removing Boc from VIII-1-Boc using TFA and was directly used for the next reaction.

[0174]

[0175] Synthesis of compound A21: Compound VIII-1 (126 mg, 0.22 mmol), compound Ⅰ-4 (130 mg, 0.44 mmol) and DIPEA (240 μL, 1.32 mmol) were dissolved in 3 mL of dimethyl sulfoxide, heated to 80°C and reacted under this condition for 4 h. The reaction was completed under LC-MS monitoring. The reaction solution was poured into water, extracted with ethyl acetate, and separated by column chromatography (DCM:MeOH=100:1~30:1) to obtain pure brown-yellow solid powder A21 (40 mg, yield: 25%). 1 H NMR(500MHz,DMSO-d6)δ13.45(s,1H),11.10(s,1H),10.90(s,1H),9.19(s,1H),7.77(dd,J=9.3,2.6Hz,1H),7. 73(s,1H),7.71–7.60(m,2H),7.33(d,J=2.3Hz,1H),7.25(dd,J=8.7,2.3Hz,1H),6.94(td,J=9.1,2.6Hz,1H),6. 84(dd,J=8.5,4.6Hz,1H),5.08(dd,J=12.8,5.4Hz,1H),4.08(d,J=12.7Hz,2H),3.11(s,2H),3.04–2.81(m,4H), 2.67–2.51(m,10H),2.27(s,3H),2.04–1.98(m,1H),1.87(d,J=12.2Hz,2H),1.46(qd,J=12.2,11.8,3.6Hz,2H).13 C NMR(126MHz,DMSO-d6)δ173.28,170.58,169.85,168.07,167.71,167.42,157.71,1 55.21,135.17,134.49,127.57,125.69,125.46,124.30,120.76,119.25,118.11,11 8.05,115.74,112.83,110.48,108.25,106.52,106.31,61.47,60.98,60.23,53.74, 51.19,49.21(2C),47.05,31.45,27.73(2C),22.66,14.56,9.02.HR-MS(ESI):calcd for C 38 H 39 FN8O6[M+H] + :723.3049; found:723.3039.

[0176] Embodiment 22

[0177] Synthesis of (Z)-2-(4-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl)azetidin-3-yl)piperazin-1-yl)-N-(5-((5-fluoro-2-oxoindole-3-ylidene)methyl)-4-methyl-1H-pyrrol-3-yl)acetamide (A22)

[0178]

[0179] Synthesis of intermediate VIII-2: Compound VII (100 mg, 0.26 mmol), tert-butyl3-oxoazetidine-1-carboxylate (70 mg, 0.40 mmol) and AcOH (1 μL, 0.03 mmol) were dissolved in 4 mL of dimethyl sulfoxide, and then stirred at 25°C for 10 min. Sodium triacetoxyborohydride (80 mg, 0.40 mmol) was added, and the mixture was reacted at 25°C for 2 h. The reaction was completed after LC-MS monitoring. 20 mL of water was poured into the reaction system, and the mixture was extracted with ethyl acetate (30 mL×2). The organic phases were combined, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and vacuum removed to remove the solvent, and the crude product was slurried with (ethyl acetate: n-hexane = 5:1) to obtain product VIII-2-Boc (80 mg, yield: 57%). 1H NMR(500MHz,DMSO-d6)δ13.45(s,1H),10.89(s,1H),9.29–9.14(m,1H),7.76(dd,J=9.4,2.6Hz,1H),7 .73(d,J=2.1Hz,1H),7.64(dd,J=9.6,3.2Hz,1H),6.97–6.88(m,1H),6.84(dd,J=8.5,4.5Hz,1H),3.8 4(s, 2H), 3.66(s, 2H), 3.14(s, 2H), 3.06(dt, J=7.0,3.8Hz, 2H), 2.70(td, J=10.8,8.9,5.8Hz, 2H), 2.56(s, 4H), 2.41–2.34(m, 1H), 2.27(s, 3H), 1.38(s, 9H). VIII-2 was obtained by removing Boc from VIII-2-Boc using TFA and was directly used for the next step reaction.

[0180]

[0181] Synthesis of compound A22: Compound VIII-2 (82 mg, 0.15 mmol), compound Ⅰ-4 (82 mg, 0.30 mmol) and DIPEA (160 μL, 0.90 mmol) were dissolved in 3 mL of dimethyl sulfoxide, heated to 80°C and reacted under this condition for 4 h. The reaction was completed under LC-MS monitoring. The reaction solution was poured into water, extracted with ethyl acetate, and separated by column chromatography (DCM:MeOH=100:1~30:1) to obtain pure brown-yellow solid powder A22 (31 mg, yield: 29%). 1 H NMR(500MHz,DMSO-d6)δ13.45(s,1H),11.10(s,1H),10.90(s,1H),9.21(s,1H),7.77(dd,J=9.4,2.7Hz,1 H),7.74(s,1H),7.71–7.62(m,2H),6.94(td,J=9.0,2.6Hz,1H),6.86–6.81(m,1H),6.82–6.78(m,1H),6. 65(dd,J=8.5,2.1Hz,1H),5.07(dd,J=12.8,5.4Hz,1H),4.11(t,J=7.8Hz,2H),3.86(dd,J=8.7,4.9Hz,2H ),3.16(s,2H),2.91–2.84(m,1H),2.77–2.51(m,8H),2.48–2.40(m,2H),2.28(s,3H),2.12–1.97(m,2H). 13C NMR(126MHz,DMSO-d6)δ173.28,170.59,169.86,167.93,167.62,159.57,157.7 1,155.38,135.17,134.29,127.57,125.62,125.31,124.31,120.82,119.30,11 7.32,115.75,114.64,112.83,110.48,106.52,106.32,104.94,61.36,60.23,5 5.55,54.60,52.97,49.59,49.17,31.45,22.67,14.56,9.04.HR-MS(ESI):calcd for C 36 H 35 FN8O6[M+H] + :695.2736; found:695.2726.

[0182] Embodiment 23

[0183] Synthesis of (Z)-2-(4-(7-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl)-7-azaspiro[3.5]non-2-yl)piperazin-1-yl)-N-(5-(5-fluoro-2-oxoindole-3-ylidene)methyl)-4-methyl-1H-pyrrol-3-yl)acetamide (A23)

[0184]

[0185] Synthesis of intermediate VIII-3: Compound VII (100 mg, 0.26 mmol), tert-butyl2-oxo-7-azaspiro[3.5]nonane-7-carboxylate (96 mg, 0.40 mmol) and AcOH (1 μL, 0.03 mmol) were dissolved in 4 mL of dimethyl sulfoxide, and then stirred at 25°C for 10 min. Sodium triacetoxyborohydride (80 mg, 0.40 mmol) was added, and the mixture was reacted at 25°C for 2 h. The reaction was completed after LC-MS monitoring. 20 mL of water was poured into the reaction system, and the mixture was extracted with ethyl acetate (30 mL×2). The organic phases were combined, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and vacuum removed to remove the solvent, and the crude product was slurried with (ethyl acetate: n-hexane = 5:1) to obtain product VIII-3-Boc (75 mg, yield: 47%). 1H NMR (500MHz, DMSO-d6) δ13.45(s,1H),10.91(s,1H),9.31(s,1H),7.77(dd,J=9.4,2.6Hz,1H),7.73(s, 1H), 7.64 (d, J=3.2Hz, 1H), 6.94 (td, J=9.4, 8.9, 2.6Hz, 1H), 6.85 (dd, J=8.4, 4.5Hz, 1H), 3.29 (q, J=5. 7Hz, 5H), 3.20(d, J=6.8Hz, 3H), 3.09–2.92(m, 2H), 2.81(s, 2H), 2.75–2.62(m, 2H), 2.28(s, 3H), 2.19–2.00(m, 4H), 1.91(s, 2H), 1.53–1.45(m, 4H), 1.39(s, 9H). VIII-3 was obtained by removing Boc from VIII-3-Boc using TFA and was directly used for the next reaction.

[0186]

[0187] Synthesis of compound A23: Compound VIII-3 (143 mg, 0.23 mmol), compound Ⅰ-4 (130 mg, 0.46 mmol) and DIPEA (250 μL, 1.38 mmol) were dissolved in 3 mL of dimethyl sulfoxide, heated to 100 ° C and reacted under this condition for 4 hours. The reaction was completed under LC-MS monitoring. The reaction solution was poured into water, extracted with ethyl acetate, and separated by column chromatography (DCM: MeOH = 100: 1 ~ 30: 1) to obtain pure brown-yellow solid powder A23 (21 mg, yield: 12%). 1 H NMR(500MHz,DMSO-d6)δ13.45(s,1H),11.10(s,1H),10.90(s,1H),9.20(s,1H),7.77(d,J=9.4Hz ,1H),7.74(s,1H),7.65(d,J=10.2Hz,2H),7.32(s,1H),7.24(d,J=8.8Hz,1H),6.94(t,J=9.0Hz, 1H),6.84(dd,J=8.6,4.7Hz,1H),5.07(dd,J=12.7,5.4Hz,1H),3.47(d,J=6.4Hz,4H),3.14(s,2H ),2.89(t,J=13.5Hz,1H),2.67–2.51(m,8H),2.27(s,3H),2.04–1.95(m,4H),1.70–1.45(m,8H). 13C NMR(126MHz,DMSO-d6)δ173.29,170.60,169.86,168.09,167.69,167.41,157.71,155.36 ,135.17,134.50,127.57,125.70,125.46,124.31,120.65,119.20,118.08,117.86,115. 76,113.03,110.48,108.23,106.52,106.33,61.43,60.24,54.89,53.03,49.78,49.19(2 C),45.08,44.86,36.68,31.61(2C),31.45,29.52,22.66,14.45,9.00.HR-MS(ESI):calcd for C 41 H 43 FN8O6[M+H] + :763.3362; found:763.3351.

[0188] Embodiment 24

[0189] Synthesis of (Z)-2-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl)azetidin-3-yl)methyl)piperazin-1-yl)-N-(5-((5-fluoro-2-oxoindole-3-ylidene)methyl)-4-methyl-1H-pyrrol-3-yl)acetamide (A24)

[0190]

[0191] Synthesis of intermediate VIII-4: Compound VII (100 mg, 0.26 mmol), tert-butyl3-formylazetidine-1-carboxylate (75 mg, 0.40 mmol) and AcOH (1 μL, 0.03 mmol) were dissolved in 4 mL of dimethyl sulfoxide, and then stirred at 25°C for 10 min. Sodium triacetoxyborohydride (80 mg, 0.40 mmol) was added, and the mixture was reacted at 25°C for 2 h. The reaction was completed after LC-MS monitoring. 20 mL of water was poured into the reaction system, and the mixture was extracted with ethyl acetate (30 mL×2). The organic phases were combined, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and vacuum removed to remove the solvent, and the crude product was slurried with (ethyl acetate: n-hexane = 5:1) to obtain the product VIII-4-Boc (95 mg, yield: 66%). 1H NMR(500MHz,DMSO-d6)δ13.45(s,1H),10.91(s,1H),9.27(s,1H),7.77(dd,J=9.3,2.6Hz,1H),7.7 4(s,1H),7.65(d,J=3.1Hz,1H),6.94(td,J=9.5,9.1,2.6Hz,1H),6.85(dd,J=8.4,4.5Hz,1H),3.9 5–3.79 (m, 4H), 3.65–3.51 (m, 4H), 3.24 (dt, J=19.8, 6.6 Hz, 2H), 2.69–2.55 (m, 4H), 2.28 (s, 2H), 1.92 (s, 1H), 1.38 (s, 9H), 1.25 (q, J=4.1, 2.8 Hz, 2H). VIII-4 was obtained by removing Boc from VIII-4-Boc using TFA and was directly used for the next reaction.

[0192]

[0193] Synthesis of compound A23: Compound VIII-4 (147 mg, 0.25 mmol), compound Ⅰ-4 (140 mg, 0.50 mmol) and DIPEA (310 μL, 1.75 mmol) were dissolved in 4 mL of dimethyl sulfoxide, heated to 80°C and reacted under this condition for 4 h. The reaction was completed under LC-MS monitoring. The reaction solution was poured into water, extracted with ethyl acetate, and separated by column chromatography (DCM:MeOH=100:1~30:1) to obtain pure brown-yellow solid powder A24 (28 mg, yield: 16%). 1 H NMR(500MHz,DMSO-d6)δ13.45(s,1H),11.09(s,1H),10.90(s,1H),9.20(s,1H),7.77(dd,J=9.4,2.6Hz,1H ),7.74(s,1H),7.70–7.61(m,2H),6.93(td,J=9.1,2.7Hz,1H),6.84(dd,J=8.4,4.4Hz,1H),6.77(d,J=2.1 Hz,1H),6.64(dd,J=8.4,2.1Hz,1H),5.06(dd,J=12.7,5.5Hz,1H),4.13(t,J=8.1Hz,2H),3.73–3.66(m,2H ),3.14(s,2H),3.09–2.94(m,2H),2.94–2.81(m,2H),2.70–2.52(m,10H),2.28(s,3H),2.04–2.00(m,1H). 13C NMR(126MHz,DMSO-d6)δ173.28,170.59,169.86,167.95,167.64,159.57,157.71,1 55.62,135.17,134.27,127.57,125.69,125.26,124.30,120.73,119.23,117.11,1 15.77,114.49,113.02,112.83,110.48,106.52,106.32,62.19,61.40,60.23,56.1 4,55.39,53.33,49.16,31.45,27.49,22.68,21.24,14.56,9.02.HR-MS(ESI):calcd for C 37 H 37 FN8O6[M+H] + :709.2893; found:709.2894.

[0194] Embodiment 25

[0195] Synthesis of (Z)-2-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-6-fluoro-1,3-dioxoisoindol-5-yl)piperidin-4-yl)methyl)piperazin-1-yl)-N-(5-((5-fluoro-2-oxoindole-3-ylidene)methyl)-4-methyl-1H-pyrrol-3-yl)acetamide (A25)

[0196]

[0197] Synthesis of intermediate II-15: Compound I-5 (180 mg, 0.62 mmol), piperidin-4-ylmethanol (85 mg, 0.74 mmol) and DIPEA (500 μL, 3.10 mmol) were dissolved in 5 mL of dimethyl sulfoxide, the reaction system was heated to 80 ° C, and reacted under this condition for 4 hours. The reaction was completed under LC-MS monitoring. The reaction system was cooled to room temperature and then poured into water, extracted with ethyl acetate, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain II-15 (233 mg, yield: 96%).

[0198]

[0199] Synthesis of intermediate III-11: II-15 (233 mg, 0.60 mmol) was dissolved in 10 mL of dichloromethane, and then DMP (507 mg, 1.20 mmol) was slowly added. The reaction was allowed to react for 4 h at room temperature. After the reaction was completed by LC-MS monitoring, an excess of saturated aqueous sodium thiosulfate and saturated aqueous sodium bicarbonate were added to the reaction system, and the mixture was stirred for another 10 min. The mixture was extracted with dichloromethane, washed with water, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the solvent was removed in vacuo. The mixture was then separated by column chromatography (DCM: MeOH = 100: 1 to 20: 1) to obtain product III-11 (106 mg, yield: 46%). HR-MS (ESI): calculated for C 19 H 18 FN3O5[M+H] + :388.12; found:388.13.

[0200]

[0201] Synthesis of compound A25: Compound III-11 (106 mg, 2.73 mmol), compound VII (87 mg, 2.27 mmol) and acetic acid (13 μL, 2.27 mmol) were dissolved in 5 mL of N,N-dimethylformamide, stirred at room temperature for 20 min, and then sodium triacetoxyborohydride (58 mg, 2.73 mmol) was slowly added, and the mixture was reacted at room temperature. The reaction was completed after LC-MS monitoring. The reaction solution was poured into water, extracted with ethyl acetate, and separated by column chromatography (DCM:MeOH=100:1~30:1) to obtain pure orange-yellow solid powder A25 (45 mg, yield: 26%). 11H NMR (500 MHz, DMSO-d6) δ 13.44 (s, 1H), 11.12 (s, 1H), 10.89 (s, 1H), 9.19 (s, 1H), 7.77 (dd, J = 9.3, 2.6 Hz, 1H), 7.74–7.69 (m, 2H), 7.66 (d, J = 3.1 Hz, 1H), 7.44 (d, J = 7.4 Hz, 1H), 6.94 (ddd, J = 9.6, 8.4, 2.6 Hz, 1H), 6.84 (dd, J = 8.5, 4.5 Hz, 1H), 5.11 (dd, J = 12.9, 5.4 Hz, 1H), 3.61 (d, J = 11.2 Hz, 2H), 3.13 (s, 2H), 2.94–2.85 (m, 3H), 2.62–2.51 (m, 6H), 2.48 (d, J = 4.4 Hz, 3H), 2.28 (s, 3H), 2.22 (d, J = 7.1 Hz, 2H), 2.07–2.01 (m, 1H), 1.83 (d, J = 12.4 Hz, 3H), 1.32–1.21 (m, 3H). 13 13C NMR (126 MHz, DMSO-d6) δ 173.23, 170.39, 169.87, 167.69, 167.18, 166.71, 166.69, 159.59, 158.72, 157.73, 156.70, 146.38, 146.31, 135.19, 129.27, 129.26, 127.58, 127.50, 125.68, 125.62, 124.32, 123.29, 123.21, 120.57, 119.12, 115.79, 115.76, 114.14, 113.02, 112.83, 112.44, 112.24, 110.48, 110.41, 106.52, 106.31, 64.22, 61.46, 53.79, 53.43, 50.40, 49.51, 32.75, 31.43, 30.70, 22.56, 8.97. HR-MS (ESI): calcd for C 39 H 40 F2N8O6 [M + H] + : 755.30; found: 755.31.

[0202] Example 26

[0203] Synthesis of (Z)-2-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-6-fluoro-1,3-dioxoisoindol-5-yl)piperidin-4-yl)methyl)piperazin-1-yl)-N-(5-((5-fluoro-2-oxoindole-3-ylidene)methyl)-2,4-dimethyl-1H-pyrrol-3-yl)acetamide (A26)

[0204]

[0205] Synthesis of intermediate Ⅹ: Compound Ⅸ (320 mg, 0.90 mmol) and 1-tert-butyloxycarbonylpiperazine (860 mg, 3.9 mmol) were dissolved in N,N-dimethylformamide (10 mL), and the temperature was raised to 50 °C for reaction for 12 h. The reaction was completed under LC-MS monitoring. 60 mL of water was added to the reaction system, and a solid precipitated, namely the product, which was filtered and dried to obtain a brown-yellow solid Ⅹ (140 mg, yield: 31%).

[0206]

[0207] Synthesis of intermediate XI: Compound X (140 mg, 0.28 mmol) was dissolved in a small amount of a mixture of dichloromethane and methanol, and then a hydrochloric acid ethyl acetate solution was added, and the mixture was sealed and reacted at room temperature. The reaction was monitored by TLC, and the solvent was removed by vacuum concentration, and then stirred with ethyl acetate (10 mL) for 0.5 h, and the ethyl acetate was removed by vacuum concentration. Finally, a large amount of ethyl acetate was used for slurrying, and the filter cake was washed with a large amount of ethyl acetate, and then washed with a small amount of n-hexane, and the filter cake was pulled dry by an oil pump. A brown-yellow compound XI (74 mg, yield: 66%) was obtained.

[0208]

[0209] Synthesis of compound A26: Compound III-10 (60 mg, 0.22 mmol), compound XI (74 mg, 0.19 mmol) and acetic acid (11 μL, 0.19 mmol) were dissolved in 5 mL of N,N-dimethylformamide, stirred at room temperature for 20 min, and then sodium triacetoxyborohydride (50 mg, 0.195 0.22 mmol) was slowly added, and then reacted at room temperature. The reaction was completed under LC-MS monitoring. The reaction solution was poured into water, extracted with ethyl acetate, and separated by column chromatography (DCM:MeOH=100:1~30:1) to obtain pure brown-yellow solid powder A26 (40 mg, yield: 29%). 1H NMR(500MHz,DMSO-d6)δ13.61(s,1H),11.08(s,1H),10.82(s,1H),9.01(d,J=31.9Hz,1H),7.73–7.6 1(m,3H),7.32(s,1H),7.23(s,1H),6.93–6.87(m,1H),6.84(dd,J=8.4,4.6Hz,1H),5.07(dd,J=12.8, 5.4Hz,1H),4.05(d,J=12.5Hz,2H),3.15(s,2H),2.97(t,J=12.2Hz,2H),2.87(d,J=16.8Hz,1H),2.6 9–2.52(m,7H),2.19(d,J=14.8Hz,7H),2.04–1.98(m,1H),1.81(d,J=13.1Hz,2H),1.51–1.00(m,7H). 13 C NMR(126MHz,DMSO-d6)δ173.27,170.58,169.97,168.11,167.44,157.71,155.46,134.71 ,134.52,132.72,131.14,129.98,128.04,127.94,127.86,125.48,124.94,122.06,118.0 7,117.85,113.38,112.31,112.12,110.34,110.27,108.22,106.02,105.81,53.28,49.21 ,47.68,32.01,31.62,31.46,29.99,29.90,29.48,22.67,12.31,9.65.HR-MS(ESI):calcd for C 40 H 43 FN8O6[M+H] + :751.33; found:751.34.

[0210] Embodiment 27

[0211] Synthesis of (Z)-2-(2,6-dioxopiperidin-3-yl)-5-(4-((4-(5-fluoro-2-oxoindole-3-ylidene)methyl)-2,4-dimethyl-1H-pyrrole-3-carbonyl)piperazin-1-yl)methyl)piperidin-1-yl)isoindole-1,3-dione (A27)

[0212]

[0213] Synthesis of intermediate ⅫⅠ-1: Compound Ⅻ (1.65 g, 5.63 mmol) was dissolved in 8 mL N,N-dimethylformamide, DIPEA (1.5 mL, 8.45 mmol) and HATU (2.14 g, 5.63 mmol) were added thereto, and the system was stirred at room temperature for 20 min, and then a DMF solution of N-Boc piperazine (1.05 g, 5.63 mmol) was added to the system, and then the system was reacted at room temperature for 12 h. After the reaction was completed under LC-MS monitoring, 40 mL of water was added to the reaction system to precipitate a solid, which was filtered and dried to obtain a yellow powder XII-1-Boc (1.57 g, yield: 67%). Compound XIII-1-Boc (1.50 g) was dissolved in a mixed system of a small amount of dichloromethane and methanol, and then a hydrochloric acid ethyl acetate solution was added, and the mixture was sealed and reacted at room temperature. After the reaction was completed under TLC monitoring, the solvent was removed by vacuum concentration, and then ethyl acetate (10 mL) was used for stirring for 0.5 h, and ethyl acetate was removed by vacuum concentration. Finally, a large amount of ethyl acetate was used for slurrying, and the filter cake was washed with a large amount of ethyl acetate, and then washed with a small amount of n-hexane, and then pulled dry with an oil pump. The brown-yellow compound XIII-1 (0.80 g, yield: 68%) was obtained. 1 H NMR (500MHz, DMSO-d6) δ13.69(s,1H),10.95(s,1H),7.78(dd,J=9.4,2.6Hz,1H),7.73(s,1H),6. 95(td,J=9.5,9.0,2.6Hz,1H),6.86(dd,J=8.4,4.5Hz,1H),3.69(s,4H),2.32(d,J=16.5Hz,6H).

[0214]

[0215] Synthesis of compound A27: Compound III-10 (950 mg, 2.61 mmol), compound XIII-1 (800 mg, 2.17 mmol) and acetic acid (125 μL, 2.17 mmol) were dissolved in 5 mL of N,N-dimethylformamide, and 5 mL of triethylamine was added to the system. The mixture was stirred at room temperature for 30 min, and then sodium triacetoxyborohydride (550 mg, 2.61 mmol) was slowly added. The mixture was reacted at room temperature for 12 h. The reaction was completed after LC-MS monitoring. The reaction solution was poured into water, extracted with ethyl acetate, and separated by column chromatography (DCM:MeOH=100:1~30:1) to obtain pure brown-yellow solid powder A27 (400 mg, yield: 26%). 1H NMR(500MHz,DMSO-d6)δ13.64(s,1H),11.10(s,1H),10.90(s,1H),7.76(dd, J=9.3,2.6Hz,1H),7.71(s,1H),7.65(d,J=8.5Hz,1H),7.31(d,J=2.3Hz,1H), 7.23(dd,J=8.9,2.4Hz,1H),6.92(dd,J=9.2,2.5Hz,1H),6.85(dd,J=8.5,4.5 Hz,1H),5.07(dd,J=12.8,5.5Hz,1H),2.55–1.74(m,25H),1.36–1.04(m,5H). 13 C NMR (126MHz, DMSO-d6) δ173.29,170.82,170.59,170.02,168.10,167.43,159.63,157.77,155.41,135.06,134.50,127.62,126.51,125.47,125 .25,118.10,113.00,110.53,108.27,106.51,106.31,60.23,49.21,47. 61,31.46,29.98,22.66,21.24,14.56,13.00,10.58.HR-MS(ESI):calcd for C 39 H 40 FN7O6[M+H] + :722.30; found:722.31.

[0216] Embodiment 28

[0217] Synthesis of (Z)-N-(2-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl)piperidin-4-yl)methyl)piperazin-1-yl)ethyl)-5-((5-fluoro-2-oxoindole-3-ylidene)methyl)-2,4-dimethyl-1H-pyrrole-3-carboxamide (A28)

[0218]

[0219] Synthesis of intermediate ⅫⅠ-2: Compound Ⅻ (300 mg, 1.00 mmol) was dissolved in 6 mL of N,N-dimethylformamide, and DIPEA (261 μL, 1.50 mmol) and HATU (380 mg, 1.00 mmol) were added thereto. After stirring at room temperature for 20 min, a DMF solution of 4-N-(2-aminoethyl)-1-N-BOC-piperazine (230 mg, 1.00 mmol) was added to the system. The system was then reacted at room temperature for 12 h. The reaction was completed after LC-MS monitoring. 40 mL of water was added to the reaction system to precipitate a solid, which was filtered and dried to obtain a yellow solid. Powdered ⅫⅠ-2-Boc (125 mg, yield: 25%) and compound ⅩIII-2-Boc (120 mg) were dissolved in a small amount of a mixed system of dichloromethane and methanol, and then hydrochloric acid ethyl acetate solution was added, sealed, and reacted at room temperature. The reaction was monitored by TLC, and the solvent was removed by vacuum concentration, and then stirred with ethyl acetate (10 mL) for 0.5 h, and the ethyl acetate was removed by vacuum concentration. Finally, a large amount of ethyl acetate was used for slurrying, and suction filtration was performed. The filter cake was washed with a large amount of ethyl acetate, and then washed with a small amount of n-hexane, and pulled dry by oil pump. The brown-yellow compound ⅪII-2 (65 mg, yield: 64%) was obtained. 1 H NMR(500MHz,DMSO-d6)δ13.75(s,1H),10.96(s,1H),7.85–7.64(m,2H),6.94(td, J=9.0,2.5Hz,1H),6.86(dd,J=8.5,4.4Hz,1H),4.02–3.24(m,16H),2.48(s,3H).

[0220]

[0221] Synthesis of compound A28: Compound III-10 (70 mg, 0.18 mmol), compound XIII-2 (65 mg, 0.15 mmol) and acetic acid (20 μL, 0.15 mmol) were dissolved in 5 mL of N,N-dimethylformamide, and 3 mL of triethylamine was added to the system. The mixture was stirred at room temperature for 30 min, and then sodium triacetoxyborohydride (50 mg, 0.18 mmol) was slowly added. The mixture was reacted at room temperature for 12 h. The reaction was completed after LC-MS monitoring. The reaction solution was poured into water, extracted with ethyl acetate, and separated by column chromatography (DCM:MeOH=100:1~30:1) to obtain pure brown-yellow solid powder A28 (35 mg, yield: 30%). 1H NMR (500MHz, DMSO-d6) δ13.69(s,1H),11.09(s,1H),10.90(s,1H),7.82–7.72(m,2H),7.65(d,J=8.5Hz,1H),7. 46(d,J=5.8Hz,1H),7.31(d,J=2.3Hz,1H),7.23(dd,J=8.7,2.3Hz,1H),6.96–6.90(m,1H),6.85(dd,J=8.4,4.5 Hz,1H),5.07(dd,J=12.8,5.4Hz,1H),4.04(d,J=12.6Hz,2H),3.00–2.87(m,3H),2.62–2.51(m,6H),2.50(s,3H ),2.45(d,J=7.4Hz,9H),2.14(s,2H),2.04–1.98(m,1H),1.79(d,J=12.8Hz,3H),1.14(q,J=13.4,12.0Hz,2H). 13 CNMR(126MHz,DMSO-d6)δ173.28,170.59,170.04,168.11,167.44,164.93,159.63,157.77,155. 47,137.09,134.98,134.51,130.64,127.68,127.60,126.28,125.46,125.35,121.21,118.02,1 17.80,115.09,115.07,112.93,112.73,110.51,110.44,108.17,106.50,106.29,64.21,57.22, 53.70,53.10,49.20,47.70,36.54,32.90,31.46,30.07,22.67,13.81,11.10.HR-MS(ESI):calcd for C 41 H 45 FN8O6[M+H] + :765.34; found:765.35.

[0222] Embodiment 29

[0223] Synthesis of (Z)-N-(3-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl)piperidin-4-yl)methyl)piperazin-1-yl)propyl)-5-((5-fluoro-2-oxoindole-3-ylidene)methyl)-2,4-dimethyl-1H-pyrrole-3-carboxamide (A29)

[0224]

[0225] Synthesis of intermediate ⅫⅠ-3: Compound Ⅻ (300 mg, 1.00 mmol) was dissolved in 6 mL of N,N-dimethylformamide, and DIPEA (261 μL, 1.50 mmol) and HATU (380 mg, 1.00 mmol) were added thereto. After stirring at room temperature for 20 min, a DMF solution of 4-N-(2-aminoethyl)-1-N-BOC-piperazine (230 mg, 1.00 mmol) was added to the system. The system was then reacted at room temperature for 12 h. The reaction was completed after LC-MS monitoring. 40 mL of water was added to the reaction system to precipitate a solid, which was filtered and dried to obtain a yellow solid. Powdered ⅫⅠ-3-Boc (200 mg, yield: 37%) compound ⅩIII-3-Boc (200 mg) was dissolved in a small amount of a mixed system of dichloromethane and methanol, and then a hydrochloric acid ethyl acetate solution was added, sealed, and reacted at room temperature. The reaction was monitored by TLC, and the solvent was removed by vacuum concentration, and then stirred with ethyl acetate (10 mL) for 0.5 h, and the ethyl acetate was removed by vacuum concentration. Finally, a large amount of ethyl acetate was used for slurrying, and suction filtration was performed. The filter cake was washed with a large amount of ethyl acetate, and then washed with a small amount of n-hexane, and pulled dry by oil pump. The brown-yellow compound ⅪII-3 (120 mg, yield: 74%) was obtained. 1 H NMR (500MHz, DMSO-d6) δ13.72(s,1H),10.95(s,1H),7.78(dd,J=9.4,2.6Hz,1H),7.73(s,1H),6.96–6.91(m,1H),6.86(dd,J=8.4,4.5Hz,1H),3.70 (d,J=12.9Hz,1H),3.53(s,1H),3.48–3.41(m,2H),3.25–3.06(m,3H),2.9 8(d,J=9.0Hz,1H),2.45(d,J=9.7Hz,6H),2.03–1.91(m,3H),1.42(s,4H).

[0226]

[0227] Synthesis of compound A29: Compound III-10 (130 mg, 0.18 mmol), compound XIII-3 (120 mg, 0.15 mmol) and acetic acid (17 μL, 0.15 mmol) were dissolved in 5 mL of N,N-dimethylformamide, and 3 mL of triethylamine was added to the system. The mixture was stirred at room temperature for 30 min, and then sodium triacetoxyborohydride (74 mg, 0.18 mmol) was slowly added. The mixture was reacted at room temperature for 12 h. The reaction was completed after LC-MS monitoring. The reaction solution was poured into water, extracted with ethyl acetate, and separated by column chromatography (DCM: MeOH = 100: 1 ~ 30: 1) to obtain pure brown-yellow solid powder A29 (130 mg, yield: 57%). 1 H NMR (500MHz, DMSO-d6) δ13.68(s,1H),11.09(s,1H),10.90(s,1H),7.77(dd,J=9.4,2.6Hz,1H),7.72(s,1H),7.65( d,J=8.4Hz,1H),7.30(s,1H),7.22(d,J=8.7Hz,1H),6.93(td,J=9.0,2.6Hz,1H),6.85(dd,J=8.4,4.5Hz,1H),5.07 (dd,J=12.8,5.4Hz,1H),4.04(s,2H),3.25(q,J=6.6Hz,2H),2.98–2.85(m,3H),2.62–2.51(m,4H),2.50(s,2H),2. 43(d,J=8.8Hz,13H),2.12(s,2H),2.04–1.98(m,1H),1.77(d,J=13.1Hz,3H),1.67(s,2H),1.13(d,J=13.5Hz,2H). 13 C NMR (126MHz, DMSO-d6) δ173.29,170.60,170.04,168.11,167.44,165.19,159.63,157. 77,155.45,136.83,134.97,134.50,130.67,127.67,127.60,126.26,125.46,125.37, 121.47,118.03,117.81,115.01,112.93,112.73,110.52,110.45,108.19,106.50,106 .29,49.19,47.68,37.54,32.87,31.45,30.02,22.66,13.79,11.04.HR-MS(ESI):calcd forC 41 H 45 FN8O6[M+H]+ :778.36; found:778.36.

[0228] Embodiment 30

[0229] Synthesis of (Z)-2-(4-((1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl)piperidin-4-yl)methyl)piperazin-1-yl)-N-(5-((5-fluoro-2-oxoindole-3-ylidene)methyl)-4-methyl-1H-pyrrol-3-yl)acetamide hydrochloride (A20.HCl)

[0230]

[0231] Synthesis of Compound A20 hydrochloride: Compound A20 (0.05 g, 0.07 mmol) was dissolved in a small amount of a mixture of dichloromethane and methanol, and then a hydrochloric acid ethyl acetate solution was added, sealed, reacted at room temperature, and the reaction was completed by TLC monitoring, vacuum concentration was performed to remove the solvent, and then ethyl acetate (10 mL) was used to stir for 0.5 h, and ethyl acetate was removed by vacuum concentration, and finally a large amount of ethyl acetate was used for slurrying, and suction filtration was performed. The filter cake was washed with a large amount of ethyl acetate, and then washed with a small amount of n-hexane, and pulled dry by oil pump. Red-brown compound A20.HCl (0.04 g, yield: 73%) was obtained. 1 H NMR(500MHz,DMSO-d6)δ13.54(s,1H),11.08(s,1H),10.94(s,1H),10.34(s,1H),7.77(dd,J=9.3,2.6Hz,1H),7.75–7.69 (m,2H),7.67(d,J=8.5Hz,1H),7.36(d,J=2.3Hz,1H),7.28(dd,J=8.7,2.3Hz,1H),6.95(td,J=9.1,2.6Hz,1H),6.86(dd,J =8.4,4.5Hz,1H),5.07(dd,J=12.8,5.4Hz,1H),4.31(s,2H),3.83–3.74(m,6H),3.45(s,2H),3.13(s,2H),3.00(t,J=12. 0Hz,2H),2.93–2.86(m,1H),2.62–2.51(m,4H),2.35(s,3H),2.18(s,1H),2.05–1.97(m,3H),1.29(q,J=10.5,8.9Hz,2H).

[0232] Embodiment 31

[0233] Proliferation inhibitory activity of compounds on human AML cell lines

[0234] Experimental method: Human acute myeloid leukemia MV-4-11 and MOLM-13 are FLT3 mutant cell lines. The CCK-8 method was used to determine the in vitro antiproliferative activity of the compound against MV-4-11 and MOLM-13: dilute the test compound with a 2-fold gradient in the culture medium to twice the final concentration, and take 200 μL to a 2mL EP tube for later use. Take an appropriate amount of cells in the logarithmic growth phase and resuspend them in the culture medium, add an equal volume to the culture medium containing the test compound, turn it upside down 10 times to mix, and add it to a 96-well plate in sequence, 100 μL per well. After culturing in an incubator at 37°C and 5% CO2 for 48 hours, add 10 μL of CCK-8 to each well and continue incubating for 2 hours. The OD450 absorbance value was read with an enzyme reader, and the experiment was repeated three times. Graphpad Prism 5 software was used to analyze and process the data to obtain the IC 50 .

[0235] The results showed that alkyl linkers were superior to polyethylene glycol (PEG) and alkylamide linkers. Compounds with 8 or 9-atom-long linker chains had stronger antiproliferative activity against human AML cells. Compounds with heterocyclic rigid chains had significantly improved activity. Among them, compound A20 had the strongest antiproliferative activity against MV4-11 and MOLM-13 cells, with an IC 50 The values ​​were 39.9 nM and 169.9 nM respectively.

[0236] Table 1. Proliferation inhibition activity of Example compounds A1-A24 on human AML cells

[0237]

[0238]

[0239]

[0240] Table 2. Proliferation inhibition activity of Example compounds A25-A30 on human AML cells

[0241]

[0242]

[0243] Embodiment 32

[0244] Effects of compounds on the degradation of FLT3 protein in MV-4-11 and MOLM-13 cells

[0245] Experimental method: MV-4-11 and MOLM-13 cells in the logarithmic growth phase were treated with solvent control DMSO and 100 nM of the compound, respectively. After culturing in an incubator at 37°C and 5% CO2 for 24 h, the cells were lysed with RIPA lysis buffer to collect the protein and ultrasonicated. The expression of cellular FLT3 protein was detected by Western Blot.

[0246] The results showed that the compounds could significantly degrade FLT3 protein in MV-4-11 and MOLM-13 cells, among which A10, A19, A20, A26 and A28 showed strong FLT3 degradation activity ( Figure 2 ).

[0247] Embodiment 33

[0248] Antitumor effects of compounds A20, A26, and A28 in MV-4-11 nude mouse subcutaneous transplant tumor model

[0249] Experimental method: MV-4-11 cells were expanded and cultured in vitro. An appropriate amount of cells in the logarithmic growth phase were resuspended in a serum-free IMDM medium and Matrigel (1:1) suspension and prepared into 5×10 6 / 100μL cell suspension, inoculate 100μL cell suspension into the subcutaneous part of the left axilla of male Balb / c nude mice with a syringe; wait until the tumor volume grows to 100-200mm 3 When the experiment was completed, the animals with moderate tumor size were randomly divided into groups of 6 in each group; blank solvent control group, A20 (5 mg / kg / d) group, A26 (5 mg / kg / d) group, A28 (5 mg / kg / d) group; the drug was administered orally once a day for 12 days; during the administration period, the body weight and tumor diameter of the nude mice were measured every day; after the experiment, the mice were killed by cervical dislocation and the tumors were weighed. The calculation formula of tumor volume (TV) is: TV = 1 / 2 × a × b 2 , a represents the long diameter of the tumor; b represents the short diameter of the tumor.

[0250] The results showed that in the MV-4-11 nude mouse subcutaneous transplanted tumor model, A20, A26, and A28 were able to effectively inhibit tumor growth after continuous administration for 12 days, and had no effect on the weight of the mice. The tumor inhibition rates of the A20, A26, and A28 administration groups were 98.57%, 82.73%, and 65.07%, respectively.

[0251] Table 3. Antitumor effect of A20 in MV-4-11 nude mouse subcutaneous transplant tumor model

[0252]

[0253] *, p<0.05; **, p<0.01; ***, p<0.001 (compared with solvent control).

[0254] Embodiment 34

[0255] Antitumor effect of compound A20 in MV-4-11 nude mouse subcutaneous transplant tumor model

[0256] Experimental method: MV-4-11 cells were expanded and cultured in vitro. An appropriate amount of cells in the logarithmic growth phase were resuspended in a serum-free IMDM medium and Matrigel (1:1) suspension and prepared into 5×10 6 / 100μL cell suspension, inoculate 100μL cell suspension into the subcutaneous part of the left axilla of male Balb / c nude mice with a syringe; wait until the tumor volume grows to 100-200mm 3 When the tumor size was moderate, the animals were randomly divided into groups, with 6 animals in each group; blank solvent control group, A20 (1.25 mg / kg / d) group, A20 (2.5 mg / kg / d) group, A20 (5 mg / kg / d) group, A20 (10 mg / kg / d) group, PX-A13 (1.6 mg / kg / d) group; the drug was administered by gavage once a day for 12 days. During the administration period, the body weight and tumor diameter of nude mice were measured every day; after the experiment, the mice were killed by cervical dislocation and the tumors were weighed; the calculation formula of tumor volume (TV) is: TV = 1 / 2 × a × b 2 , a represents the long diameter of the tumor; b represents the short diameter of the tumor.

[0257] The results showed that in the MV-4-11 nude mouse subcutaneous transplant tumor model, A20 was able to inhibit tumor growth in a dose-dependent manner after continuous administration for 12 days, and had no effect on the weight of mice; the tumor inhibition rate of the low-dose group of 1.25 mg / kg / d was 51.43%, and the tumor inhibition rate at a dose of 2.5 mg / kg / d reached 84.34%, which was significantly better than the tumor inhibition rate of compound PX-A13 at the same molar dose (1.6 mg / kg / d) (39.76%); a dose of 5 mg / kg / d could cause some tumors to disappear, and a dose of 10 mg / kg / d could cause all tumors to disappear in 8 days.

[0258] Table 4. Antitumor effect of A20 in MV-4-11 nude mouse subcutaneous transplant tumor model

[0259]

[0260] *, p<0.05; **, p<0.01; ***, p<0.001 (compared with solvent control).

[0261] Embodiment 35

[0262] Proliferation inhibitory activity of A20 on MOLM-13 cells resistant to Sunitinib or Quizartinib

[0263] Experimental methods: AML cell lines resistant to FLT3 inhibitors were constructed by continuous drug induction for more than three months, among which MOLM-13-Suni-R and MOLM-13-Quiz-R were cell lines resistant to FLT3 inhibitors Sunitinib or Quizartinib; the in vitro antiproliferative activity of compound A20 against MOLM-13-Suni-R and MOLM-13-Quiz-R was determined by CCK-8 method: the test compound was diluted 2-fold with culture medium to twice the final concentration, and 200 μL was taken to a 2mL EP tube for later use; an appropriate amount of cells in the logarithmic growth phase were resuspended in culture medium, and an equal volume was added to the culture medium containing the test compound, inverted 10 times to mix, and added to 96-well plates in sequence, 100 μL per well; after culturing in an incubator at 37°C and 5% CO2 for 48 hours, 10 μL CCK-8 was added to each well and incubated for another 2 hours; the OD450 absorbance value was read by microplate reader, and the experiment was repeated three times; Graphpad Prism5 software analyzes and processes the data to obtain IC 50 .

[0264] The results showed that MOLM-13 cells developed resistance to Sunitinib or Quizartinib after drug induction, and the FLT3 inhibitor PX-A13 also showed weak antiproliferative activity against these two resistant cell lines; however, compound A20 still showed strong antiproliferative activity against Sunitinib or Quizartinib resistant cells, with IC 50 The values ​​were 147 nM and 483 nM, respectively (Table 5).

[0265] Table 5. Proliferation inhibitory activity of A20 on MOLM-13 cells resistant to Sunitinib or Quizartinib

[0266]

[0267]

[0268] The above are only preferred embodiments of the present invention, and the protection scope of the present invention is not limited to the above embodiments. All technical solutions under the concept of the present invention belong to the protection scope of the present invention. It should be pointed out that for ordinary technicians in this technical field, some improvements and modifications without departing from the principle of the present invention should be regarded as the protection scope of the present invention.

Claims

1. An indole ketone FLT3 protein degrader or a pharmaceutically acceptable salt thereof, the structure of which is shown below: 。 2. A pharmaceutical composition comprising the indolinone FLT3 protein degrader as described in claim 1 or a pharmaceutically acceptable salt thereof.

3. Use of the indolinone FLT3 protein degrading agent or a pharmaceutically acceptable salt thereof as claimed in claim 1 in the preparation of a FLT3 protein degrading agent.

4. Use of the indolinone FLT3 protein degrader or a pharmaceutically acceptable salt thereof as claimed in claim 1, or the pharmaceutical composition as claimed in claim 2, in the preparation of a medicament for treating and / or preventing diseases related to abnormal FLT3 expression.

5. Use of the indolinone FLT3 protein degrader or a pharmaceutically acceptable salt thereof as claimed in claim 1, or the pharmaceutical composition as claimed in claim 2 in the preparation of a medicament for treating acute myeloid leukemia.

6. Use of the indolinone FLT3 protein degrader or a pharmaceutically acceptable salt thereof as claimed in claim 1, or the pharmaceutical composition as claimed in claim 2, in the preparation of a drug for treating acute myeloid leukemia resistant to FLT3 inhibitors.

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