Benzazainydine bifunctional molecule derivatives and uses thereof

Abstract

The application belongs to the field of pharmaceutical chemistry, and particularly relates to benzindole bifunctional molecule derivatives and application thereof, the derivatives are benzindole bifunctional molecules or pharmaceutically acceptable salts thereof, and have a structure as shown in general formula (I). The derivatives are a kind of protein degradation targeting chimera (PROTACs) technology, and can induce degradation of BET proteins. The bifunctional molecule compounds are used in drugs for preventing and / or treating diseases related to BET proteins.

Description

Technical Field

[0001] This invention relates to the field of medicinal chemistry, specifically to a benzoindole bifunctional molecular derivative and its applications, particularly as a BET protein degradation inhibitor. Background Technology

[0002] Cancer is one of the leading causes of death worldwide. Overall, the five-year survival rate for cancer is currently low, and early detection and cure are extremely difficult. The rapid development of kinase-based anti-cancer drugs has passed its initial explosive growth phase. Due to limitations and characteristics of their targets, drug development presents many challenges, such as drug resistance and safety risks from multiple potential targets. Therefore, developing novel treatment methods has significant clinical value and research implications for cancer prevention, treatment, and therapy. Achieving cancer prevention and treatment is both challenging and urgent.

[0003] Bromodomains (BRDs) are conserved protein domains that specifically recognize acetylated lysine residues in histones. Currently, 61 bromodomains have been identified in mammals, encompassing 46 bromodomain proteins. Among these, BET (bromodomain and extra-terminal) proteins are the most representative and the most extensively studied class of proteins. BET proteins are important co-regulators of gene transcription. The BET protein family includes BRD2, BRD3, BRD4, and BRDT. BRD3 directly interacts with the transcription factor GATA1, causing GATA1 to induce the expression of known erythrocyte-specific genes, thereby inhibiting proliferation. BRD4 promotes the dissociation of the repressor HEXIM1, thereby activating p-TEFb and further phosphorylating and activating RNAPol II. Numerous studies have shown that the proto-oncogene MYC drives the development of various tumor diseases, and BET proteins can regulate the expression level of c-MYC protein in cancer cells. BRDT plays a crucial role in chromatin remodeling during spermatogenesis; therefore, BRDT plays an important role in maintaining sperm head chromatin structure. In summary, BET protein plays an important role in the physiology and pathology of the occurrence and development of human cancer.

[0004] BET proteins play a crucial role in various human diseases, including cancer, inflammation, HIV infection, and cardiovascular disease. Cancer is currently the most popular research area for BET proteins, with nearly 20 compounds currently in clinical trials. At present, there are relatively few structural types of small molecule inhibitors, with compounds in clinical trials mainly belonging to three classes: triazoles, quinolinones, and pyridone-pyrroles. These inhibitors have relatively simple structures, lack selectivity for the BET Brmodomain, and exhibit poor pharmacokinetic and in vitro properties. Furthermore, in clinical trials, high doses of these non-selective pan-BET inhibitors are poorly tolerated by patients, necessitating dose reductions at the expense of efficacy. Overall, pan-BET inhibitors have faced some setbacks in both early-stage research and clinical trials. Therefore, this presents a new opportunity for the research and development of selective BET inhibitors. In recent years, with the introduction of protein hydrolysis targeted chimeras (PROTACs) technology into the development of BET protein drugs, the new technology has brought huge advantages to the treatment methods, thus giving rise to a surge in drug research and development of protein degrading agents. In particular, ARV-110, as the first protein degrading agent to enter clinical research, has greatly encouraged the development of BET protein degrading agents. Summary of the Invention

[0005] This invention, through studying the crystal structure model of BET protein and summarizing the structure-activity relationship of BET, designed and synthesized benzoindole bifunctional molecular derivatives and their applications. Pharmacological results show that the BET protein-inhibiting compounds of this invention have good BET protein inhibitory activity, while the bifunctional molecular protein degraders can efficiently induce apoptosis and have good application prospects. Therefore, this invention develops a class of novel selective BET inhibitors and protein degraders with novel core structures to prevent and treat diseases related to this protein pathway. This invention provides a class of novel BET inhibitors and BET protein degraders with novel core structures. These inhibitors have good protein affinity and selectivity, and protein degradation based on these inhibitors exhibits excellent protein degradation induction efficiency and cell activity.

[0006] To address the problems in the existing technology, the technical solution adopted by this invention is as follows:

[0007] A bifunctional molecule having the following structure or a pharmaceutically acceptable salt thereof, having a structure as shown in general formula (II). That is, BET protein ligand-linker-E3 ubiquitination ligand, wherein the BET protein ligand is a benzoindolone compound that can bind BET protein, the E3 ubiquitination ligand is a compound that binds Cerebro or VHL protein, and the linker ALB is a structural unit that connects the two parts of BET protein ligand and E3 ubiquitination ligand.

[0008] In general formula (II) In some parts:

[0009] R 1 It represents hydrogen, methyl, ethyl, n-propyl, cyclopropyl, isopropyl, tert-butyl, n-butyl, haloalkyl, heterocycloalkyl, or cycloalkyl;

[0010] Ar represents aryl; aryl groups may be optionally substituted with the following groups: halogen, -CN, -CF3, OCF3, -OR 2 -SR 2 -NR 2 2. -COR 2 ;

[0011] A represents -Het-, -R 2 -、-COR 2 -、-CONR 2 Het-、-COR 2 Het-, -CONHHet-, -COOHet-, -NHR 2 -、-NH(CH2) 1-10 Het-, -NH(CH2) 1-10 OR 2 -、-NH(CH2) 1-10 NR 2 -、-NHCOOR 2 -、-NHCOOHet-、-NHCONHR 2 -、-NHCONHHet-、-OR 2 -、-O(CH2) 1-10 NR 2 -、-O(CH2) 1-10 Het- or -O(CH2) 1-10 OR 2 -;

[0012] R 2 Indicates alkyl, haloalkyl, heterocyclic alkyl, or cycloalkyl;

[0013] In general formula (II)

[0014] B represents O, S, NH, CONH, CH2, CH 1-10 CONH, NNHCOCH 1-10 Or NHHet;

[0015] X represents CO or CH2;

[0016] G represents H, alkyl, OH, or CH2Het;

[0017] L represents an aliphatic chain, an aromatic chain, a nonlinear chain, a chain containing 1-5 3-10 member saturated heterocycles or aromatic heterocycles, or an alkyl chain containing 1-5 heteroatoms N, S or O.

[0018] The aryl group is a carbocyclic ring containing phenyl, naphthyl, acenaphthyl, or tetrahydronaphthyl, wherein the phenyl, naphthyl, acenaphthyl, or tetrahydronaphthyl group may be optionally substituted by 1, 2, or 3 substituents, each substituent being independently selected from hydrogen, alkyl, cyano, halogen, haloalkyl, hydroxyl, mercapto, alkoxy, alkylthio, alkoxyalkyl, aralkyl, phenyl-containing carbocyclic ring, naphthyl-containing carbocyclic ring, acenaphthyl-containing carbocyclic ring, tetrahydronaphthyl-containing carbocyclic ring, or Het.

[0019] The Het is selected from monocyclic heterocycles of piperidinyl, pyrrolyl, pyrazolyl, imidazoleyl, furanyl, morpholinyl, thiophenyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridinyl, pyrimidinyl, piperazinyl, substituted piperazinyl, pyrazinyl, or pyridazinyl; or selected from quinolinyl, quinoxolinyl, indolyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzothiazolyl, benzoxisothiazolyl, benzofuranyl, benzo[[unclear text - likely a typo, should be left as is]. A bicyclic heterocycle of thienyl, 2,3-dihydrobenzo[b][1,4]dioxane or benzo[d][1,3]dioxane; each monocyclic or bicyclic heterocycle may be optionally substituted with 1, 2 or 3 substituents, each substituent being independently selected from halogen, haloalkyl, hydroxyl, alkyl, alkoxy, C3-C8 aliphatic carbocyclic ring, tetrahydropyrrolithyl, morpholinyl, alkoxymorpholinyl, piperazine, piperidinyl or alkylaminopiperidinyl;

[0020] The alkyl group is a straight-chain or branched saturated hydrocarbon group having 1-6 carbon atoms; or a cyclic saturated hydrocarbon group having 3-6 carbon atoms; or a cyclic saturated hydrocarbon group having 3-6 carbon atoms connected to a straight-chain or branched saturated hydrocarbon group having 1-6 carbon atoms.

[0021] The alkoxy group is a straight-chain or branched saturated hydrocarbon group having 1-6 carbon atoms; or a cyclic saturated hydrocarbon group having 3-6 carbon atoms; or a cyclic saturated hydrocarbon group having 3-6 carbon atoms connected to a straight-chain or branched saturated hydrocarbon group having 1-6 carbon atoms; wherein each carbon atom may optionally be substituted with oxygen.

[0022] The alkylamino group is a straight-chain or branched saturated hydrocarbon group having 1-6 carbon atoms; or a cyclic saturated hydrocarbon group having 3-6 carbon atoms; or a cyclic saturated hydrocarbon group having 3-6 carbon atoms connected to a straight-chain or branched saturated hydrocarbon group having 1-6 carbon atoms; wherein each carbon atom may optionally be substituted with nitrogen.

[0023] The halogen is selected from fluorine, chlorine or bromine.

[0024] For the above general formula, the specific substances are as follows:

[0025] N-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)ethyl)-3-(6-(((2-methoxyphenyl)sulfonylamino)-1-methyl-2-oxo-1,2-dihydrobenzo[cd]indole-4-yl)propionamide (Ⅱ-1), N-(3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)propyl)-3-(6-((2-) -Methoxyphenyl)sulfonylamino)-1-methyl-2-oxo-1,2-dihydrobenzo[cd]indol-4-yl)propionamide (II-2), N-(4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)amino)butyl)-3-(6-(((2-methoxyphenyl)sulfonylamino)-1-methyl-2-oxo-1,2-dihydrobenzo[cd]indol-4-yl)propionamide (II-3), N-(5- ((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)pentyl)-3-(6-(((2-methoxyphenyl)sulfonylamino)-1-methyl-2-oxo-1,2-dihydrobenzo[cd]indole-4-yl)propionamide (II-4), N-(6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)hexyl)-3-(6-((2-methoxyphenyl) (2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)ethyl)piperazin-1-yl)-3-oxopropyl)-1-methyl-2-oxo-1,2-dihydrobenzo[cd]indole-6-yl)-2-methoxybenzenesulfonamide (II-6) or

[0026] N-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)amino)ethoxy)ethyl)-3-

[0027] (6-((2-methoxyphenyl)sulfonamide)-1-methyl-2-oxo-1,2-dihydrobenzo[cd]indol-4-yl)propionamide (Ⅱ-7).

[0028] The structural formula is shown in the table below:

[0029]

[0030]

[0031] A pharmaceutical composition comprising an effective dose of a compound of general formula (II) as described in any one of claims 1-2, or a mixture of one or more compounds thereof, or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable excipients.

[0032] As an improvement, the pharmaceutically acceptable salts include acid addition salts formed by compounds of general formula (I) with the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid, succinic acid, fumaric acid, salicylic acid, phenylacetic acid, or mandelic acid; or acidic salts formed with inorganic bases containing basic metal cations, alkaline earth metal cations, or ammonium cations.

[0033] The use of a bifunctional molecule or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition thereof, in the preparation of a medicament for the prevention or treatment of clinical conditions related to BET protein.

[0034] The diseases associated with BET protein mentioned above include rheumatoid arthritis, gouty arthritis, melanoma, liver cancer, kidney cancer, acute leukemia, multiple myeloma, lymphoma, non-small cell lung cancer, prostate cancer, thyroid cancer, skin cancer, pancreatic cancer, ovarian cancer, breast cancer, myelodysplastic syndrome, esophageal cancer, gastrointestinal cancer, or mesothelioma.

[0035] BRD4 protein binding ability test

[0036] AlphaScreen method

[0037] 1) Experimental materials and equipment: BRD4 protein; PHERAstar FS plate reader (BMG Labtech, Germany); HEPES, NaCl, BSA, CHAPS; 384 low-volume white plate (Corning, USA); pipette tips (Axygen, USA); DMSO (Sigma, USA)

[0038] 2) Experimental Principle: The obtained compounds were all tested using the Amplified Luminous Proximity Homogeneous Assay (AlphaScreen) to detect the activity of BRD4 protein. Donor beads and acceptor beads were each coupled to two different antibody epitopes targeting the aggrecan. When the substrate aggrecan structure is intact, the paired beads are brought closer together, generating a light signal. In the presence of BRD4 protein and small molecule compounds, the intact structure of the aggrecan is disrupted, reducing the light signal intensity. This allows for the detection of the binding affinity between small molecules and BRD4 protein. This method is rapid, simple, efficient, sensitive, and reliable. Compared with traditional protease activity detection techniques (e.g., FRET-based methods), AlphaScreen has the advantage of utilizing larger molecular weight substrates, making it a particularly suitable method for testing BRD4 inhibitor activity.

[0039] 3) Experimental steps and methods: Amplified luminscent proximity homogeneous assay (AlphaScreen) method, the specific method is as follows (taking BRD4 as an example): at room temperature with a pH of 7.4, prepare a mixed buffer solution of 50mM HEPES, 100mM NaCl, 0.1% BSA and 0.05% CHAPS in each well. The ligands were serially diluted from 150 μM at a 1:2 ratio to obtain 24 concentration gradients, and 4 μl of HIS-labeled BRD4 250 nM was added to each well. After culturing the plate for 30 minutes, 4 μl of biotinylated peptide (H4K5KAc8KAc12KAc16ac) was added. After culturing for another 30 minutes, 4 μl of the donor strain coated with 25 μg / mL streptavidin and 4 μl of the acceptor strain coated with 25 μg / mL nickel chelate were added under low light. After culturing for 60 minutes under dark conditions, the light intensity was read using a PHERAstar FS plate reader (B MG Labtech, Germany). The excitation / emission wavelengths were 680 / 570 nm, respectively.

[0040]

[0041] a IC 50 Two repeated tests; b is a positive control.

[0042] Experimental studies were conducted on the binding ability of the above compounds to the BRD4 protein, and the in vitro antitumor activity of the target compounds was determined.

[0043] The inhibitory effects on leukemia cell line MV4-11, Molm-13 tumor cell line, and MDA-MB-231 triple-negative breast cancer cell line were determined using the MTT assay.

[0044] The MTT assay utilizes the presence of NADP-associated dehydrogenases in the mitochondria of living cells, which reduce exogenous MTT to insoluble blue-purple crystals (Formazan), which then deposit within the cells. Dead cells lack this ability. The purple crystals in the cells are then dissolved using dimethyl sulfoxide (DMSO) or a triple solution (10% SDS-5% isobutanol-0.01 mol / L HCl), and the OD value is measured at 570 nm using an enzyme-linked immunosorbent assay (ELISA) scanner to indirectly reflect the number of viable cells.

[0045] Specific method: The tumor cells to be tested, which are in the logarithmic growth phase, are seeded into a 96-well culture plate at a certain cell amount. After culturing for 24 hours, the screened sample is added. The cells are then cultured for another 48 hours at 37°C and 5% CO2. MTT is added and the cells are cultured for another 4 hours. The cells are then dissolved and crystallized with DMSO and detected using an enzyme-linked immunosorbent assay (ELISA) reader.

[0046] The in vitro antitumor activity of the target compound against nodular leukemia cells MV4-11 is as follows:

[0047]

[0048] Note: a The cell inhibition rate at a concentration of 1 μM was measured twice independently. b Not tested.

[0049] Western blot analysis of the target compound:

[0050] For Western blot analysis, use 2×10 6 Compounds were processed at specified concentrations for different time periods using cells / well. Cells were collected and lysed in RIPA buffer containing protease inhibitors. 20 μg of lysate was run in each lane of a PAGE-SDS assay and blotted onto a PVDF membrane. Internal controls were detected using a horseradish peroxidase-conjugated anti-GAPDH antibody. Antibodies used for immunoblotting included BRD4 (BRD4(E2A7X) Rabbit mAb #13440) and c-Myc (Myc-Tag(71D10) Rabbit mAb #2278) purchased from Cell Signaling Technology, and GAPDH (GAPDH(D16H11)) from Santa Cruz Biotechnologies. Rabbit mAb#5174).

[0051] Bioactivity test results as follows Figure 1 As shown in the figure, the information indicates that the compounds provided by this invention have the effect of inhibiting or degrading BET protein. The compounds of this invention can be used to treat various solid organ cancers, including melanoma, liver cancer, kidney cancer, lung cancer, prostate cancer, thyroid cancer, skin cancer, pancreatic cancer, ovarian cancer, breast cancer, testicular cancer, bone cancer, brain cancer, esophageal cancer, gastrointestinal cancer, soft tissue tumors, leukemia, lymphoma, etc., which can be cancers and inflammation mediated by BET protein, or cancers independent of the above mechanisms. Therefore, this invention proposes that the compounds of this invention can be used in the preparation of anticancer drugs.

[0052] In vitro BET protein activity assays show that the compounds provided by this invention possess significant BET protein binding energy or highly efficient protein degradation capabilities due to their small molecule structure. Since BET protein plays a crucial role in tumor cell growth and proliferation, and this is supported by in vitro protein inhibitory activity experiments, the compounds provided by this invention can be used in drugs for the prevention or treatment of diseases related to BET protein inhibitors or degraders, particularly in oncology drugs.

[0053] Beneficial effects:

[0054] This invention, through studying the crystal structure model of BET protein and summarizing the structure-activity relationship of BET, designs and synthesizes benzoindole bifunctional molecular derivatives, along with their preparation methods and applications. The resulting compounds that inhibit BET protein exhibit good BET protein inhibitory activity. Simultaneously, the bifunctional molecular protein degraders can efficiently induce apoptosis. Based on this novel selective BET inhibitor with a structural core, a class of protein degraders has been developed, possessing excellent ability to induce BET protein degradation, thus preventing and treating diseases related to this protein pathway, and showing promising application prospects. Attached Figure Description

[0055] Figure 1 The dependence of BRD4 and c-Myc protein levels on the concentration gradient of compound II-3 after 18 hours of treatment of MV4-11 cell line. Detailed Implementation

[0056] The structure of the compound was determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). NMR shifts (S) were measured in 10⁻¹⁰ increments. -6 The unit (ppm) is given. NMR determination was performed using a Bruker AVANCE-300 or Bruker AVANCE-400 NMR spectrometer. The solvents used were neodimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD). The internal standard was tetramethylsilane (TMS).

[0057] MS measurements were performed using a Finnigan LCQAd(ESI) mass spectrometer (manufacturer: Thermo, model: Finnigan LCQadvantage MAX).

[0058] The preparation of highly water-soluble compounds was carried out using Biotage rapid purification preparative liquid chromatography with Flash Isoleraone, and the column used was a bonded phase series rapid separation column (SW-5222-040-SP) from Changzhou Santai Company.

[0059] Thin-layer chromatography (TLC) uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates. The silica gel plates used in TLC have a size of 0.15mm to 0.2mm, and the size used for TLC separation and purification of products is 0.4mm to 0.5mm. Column chromatography generally uses Yantai Huanghai 200-300 mesh silica gel as the carrier.

[0060] The known starting materials of this invention can be synthesized using or according to methods known in the art, or can be purchased from companies such as ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, AccelaChemBio Inc, Shanghai Bid Pharmaceutical Technology Co., Ltd., Anaiji Chemical, and Darui Chemicals.

[0061] Unless otherwise specified in the examples, the reactions can be carried out under an argon or nitrogen atmosphere.

[0062] Argon or nitrogen atmosphere refers to a reaction flask connected to an argon or nitrogen gas balloon with a volume of approximately 1L.

[0063] A hydrogen atmosphere refers to a reaction vessel connected to a hydrogen balloon with a volume of approximately 1L.

[0064] The hydrogenation reaction is usually carried out under vacuum, filled with hydrogen gas, and repeated 3 times.

[0065] Unless otherwise specified in the examples, "solution" refers to an aqueous solution.

[0066] Unless otherwise specified in the examples, the reaction temperature is room temperature, which is 20℃~30℃.

[0067] The reaction process in the examples was monitored using thin-layer chromatography (TLC). The developing solvent used in the reaction, the eluent system for column chromatography used to purify the compounds, the developing solvent system for TLC, and the rapid purification preparative liquid chromatography were all described. 18C-bonded phase separation column systems include: A: dichloromethane / methanol system, B: n-hexane / ethyl acetate system, C: petroleum ether / ethyl acetate system, D: ethyl acetate / dichloromethane system, E: ethyl acetate / dichloromethane / n-hexane, F: methanol / water system, G: methanol / water / methylamine aqueous solution system, H: acetonitrile / water system. The volume ratio of the solvent is adjusted according to the polarity of the compound, and small amounts of basic or acidic reagents such as triethylamine and acetic acid can also be added for adjustment.

[0068] Example 1

[0069] 3-(6-((2-methoxyphenyl)sulfonamide)-1-methyl-2-oxo-1,2-dihydrobenzo[cd]indol-4-yl)propionic acid (I-1g)

[0070]

[0071] Step 1: 5-Bromo-1H,3H-benzo[de]isochrome-1,3-dione (I-1a)

[0072] 1,8-Naphthalenedicarboxylic anhydride (10.0 g, 50.5 mmol) and silver sulfate (7.9 g, 25.25 mmol, 0.50 equiv) were added to a concentrated sulfuric acid solution (200 mL). After stirring at room temperature for 30 minutes, liquid bromine (3.2 mL, 63.0 mmol, 1.26 equiv) was added, and the mixture was heated to 60 °C for 8–10 hours, then cooled to 20 °C. The reaction was checked for completeness by TLC. The silver bromide byproduct solid was filtered off, yielding a clear orange solution. This orange solution was added dropwise to an ice-water mixture (1 L), stirred at room temperature for 20 minutes, and filtered to obtain a grayish-white solid. The filter cake was washed with water (50 mL), then with cold ethanol (100 mL × 2), and dried in a vacuum drying oven at 60 °C to obtain a white solid (I-1a) (12.54 g, 45.5 mmol), yield: 90%. MS m / s(ESI)[M+H] + :301.13.

[0073] Step 2: 4-Bromobenzo[cd]indol-2(1H)-one (I-1b)

[0074] 5-Bromo-1H,3H-benzo[de]isocyanone-1,3-dione (I-1a) (27.59 g, 100 mmol) and hydroxylamine hydrochloride (6.9 g, 100 mmol) were added to a pyridine (200 mL) solution and refluxed at 90 °C for 2 hours. After cooling to 80 °C, p-toluenesulfonyl chloride (38.14 g, 200 mmol, 2 equiv) was added, and the mixture was refluxed at 90 °C for another 2 hours. The mixture was then cooled to room temperature, and the reaction was checked for completeness by TLC. The reaction mixture was poured into 0.50 L of water and stirred. The mixture was filtered to obtain a pale yellow filter cake, which was washed successively with 0.5 L of NaHCO3 aqueous solution and 0.5 L of water. The cake was then filtered and dried to remove excess water for the next reaction. All intermediates were dissolved in 85 mL of ethanol and 100 mL of water and stirred at room temperature. Then, 325 mL of 1.4 mol / L NaOH aqueous solution was slowly added, and the mixture was refluxed at 90 °C with stirring for 3 hours, while simultaneously distilling off the ethanol. The reaction was checked by TLC until complete. The mixture was cooled to 75°C, and 60 mL of concentrated HCl was slowly added dropwise. After the addition was complete, the mixture was cooled to room temperature, filtered, and dried to give a white solid (I-1b) (7.17 g, 29.14 mmol), yield: 29%. MS m / s (ESI) [M+H + : 247.9.

[0075] Step 3: 4-Bromo-6-nitrobenzo[cd]indole-2(1H)-one (I-1c)

[0076] 4-Bromobenzo[cd]indol-2(1H)-one (I-1b) (7 g, 28.5 mmol) was dissolved in AcOH (15 mL), and 69% HNO3 (2 mL, 42.75 mmol, 1.5 equiv) was added. The mixture was stirred at 50-65 °C for 4 hours. The reaction was checked for completeness by TLC. The mixture was filtered, the solvent was removed by vacuum distillation, and the solution was separated by silica gel column chromatography (petroleum ether: dichloromethane = 1:5) to give a yellow solid (I-1c) (6.9 g, 23.65 mmol), yield: 83%. MS m / s (ESI) [M+H] + :291.0.

[0077] Step 4: 4-Bromo-1-methyl-6-nitrobenzo[cd]indol-2(1H)-one (I-1d)

[0078] 4-Bromo-6-nitrobenzo[cd]indol-2(1H)-one (I-1c) (6.5 g, 22.26 mmol) was dissolved in anhydrous DMF (100 mL), and 60% NaH (1.34 g, 33.35 mmol, 1.5 equiv) was added. The mixture was stirred at 0 °C for 30 min. Methyl iodomethane (2.06 mL, 33.35 mmol, 1.5 equiv) was added dropwise to the reaction mixture, and the mixture was stirred at room temperature for 3 h. The reaction was monitored by TLC until complete. The reaction mixture was poured into ice water and extracted with EA. The organic layer was washed four times with H2O and saturated brine, dried over anhydrous sodium sulfate, and allowed to stand. The mixture was filtered, the solvent was removed by vacuum distillation, and the product was separated by silica gel column chromatography to give 5.52 g (Id) of a yellow solid, with a yield of 81%. MS m / s (ESI) [M+H]+: 306.9.

[0079] Step 5: (E)-3-(1-Methyl-6-nitro-2-oxo-1,2-dihydrobenzo[cd]indol-4-yl)methyl acrylate (I-1e)

[0080] Weigh 500 mg (1.63 mmol, 1 equiv) of 4-bromo-1-methyl-6-nitrobenzo[cd]indol-2(1H)-one I-1d, 18 mg (0.08 mmol, 0.05 equiv) of Pd(AcO)2, 49 mg (0.16 mmol, 0.1 equiv) of o-triphenylphosphine, and 675 mg (4.88 mmol, 3 equiv) of K2CO3, and dissolve in 40 mL of anhydrous 1,4-dioxane. Add 293 mL (4.88 mmol, 2 equiv) of methyl acrylate to the reaction mixture. Exchange the reaction system with N2 and then heat to 80 °C for 7 h. When TLC analysis showed complete conversion of the starting material, extract the mixture with AcOEt and water. Wash the organic layer with H2O and brine, dry (MgSO4), concentrate, and purify by silica gel column chromatography to give 381 mg of a yellow solid in 75% yield. [M+H] + 313.2. 1 H NMR (300MHz, DMSO-d6) δ8.96 (s, 1H), 8.70-8.56 (m, 2H), 7.98 (d, J = 16.1Hz, 1 H),7.30(d,J=8.1Hz,1H),6.98(d,J=16.1Hz,1H),3.78(s,3H),3.40(s,3H).

[0081] Step 6: Methyl 3-(6-((2-methoxyphenyl)sulfonamide)-1-methyl-2-oxo-1,2-dihydrobenzo[cd]indol-4-yl)propionate (Ⅰ-2)

[0082] Weigh 380 mg (1.21 mmol, 1 equiv) of I-1e and dissolve it in 20 mL of ethanol. Add 100 mg of Raney-Nickel catalyst (containing approximately 50% water). Purify the reaction mixture three times with argon and twice with hydrogen. Stir for 5 h under a hydrogen atmosphere until complete reaction is detected by LTC. Filter the reaction mixture through diatomaceous earth and concentrate under reduced pressure to obtain a brown crude product for the next step. Dissolve the brown crude product in 20 mL of anhydrous DCM and add 301 mg (1.46 mmol, 1.2 equiv) of 2-methoxybenzenesulfonyl chloride and 0.5 mL of pyridine. Stir overnight at room temperature. Detect complete reaction by TLC and concentrate under reduced pressure. Purify by column chromatography (30% EA / PE) to obtain 287 mg of a yellow solid, with a yield of 53%. 1 H NMR(300MHz,Chloroform-d)δ7.99(s,1H),7.90(s,1H),7.68(dd,J=7.7,1.7Hz,1H),7.51(dd,J=15.9,1.8Hz,1H),7.16(s,1H),7.10(d,J=7 .8Hz,2H),6.95(t,J=7.7Hz,1H),6.63(d,J=7.5Hz,1H),4.13(s,3H),3.68(s,3H),3.34(s,3H),3.19(t,J=7.6Hz,2H),2.71(t,J=7.6Hz,2H).

[0083] Step 7: 3-(6-((2-methoxyphenyl)sulfonamide)-1-methyl-2-oxo-1,2-dihydrobenzo[cd]indol-4-yl)propionic acid (I-1g)

[0084] Weigh 287 mg of I-2 and dissolve it in 20 mL of methanol in a 100 mL round-bottom flask. Add 5 mL of 2 mol / L NaOH solution and stir at room temperature for 3 h. The reaction is confirmed to be complete by TCL plate testing. Remove the organic solvent by rotary evaporation under reduced pressure. Extract with EA and water, and combine the aqueous phases. Neutralize the reaction to acidity with dilute hydrochloric acid, and extract again with EA and water. Combine the organic phases, rotary evaporate under reduced pressure, and dry to obtain 255 mg of yellow solid, with a yield of 92%.

[0085] Example 2

[0086] N-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)ethyl)-3-(6-((2-methoxyphenyl)sulfonylamino)-1-methyl-2-oxo-1,2-dihydrobenzo[cd]indole-4-yl)propionamide (II-1)

[0087]

[0088] Step 1: 2-(2,6-dioxopiperidin-3-yl)-4-fluoroisoindole-1,3-dione (II-1a)

[0089] Weigh 2 g (12.04 mmol, 1 equiv) of 4-fluoroisobenzofuran-1,3-dione into a round-bottom flask, dissolve in 20 mL of AcOH, and add 2.18 g (13.24 mmol, 1.1 equiv) of 3-aminopiperidine-2,6-dione hydrochloride and AcOK (2.36 g, 24.08 mmol, 2 equiv). Stir and heat at 60 °C for 12 hours. The reaction was monitored by TLC until complete. Extraction was performed using EA, the organic layer was washed with saturated brine, dried over Mg2SO4, and the solvent was removed by vacuum distillation. Purification was performed by silica gel column chromatography to give a white solid (Ⅱ-1a) (2.13 g, 7.71 mmol), yield: 64%. MS m / s (ESI) [M+H] + :277.1.

[0090] Step 2: (2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)ethyl)tert-butyl carbamate (II-1b)

[0091] Weigh 2-(2,6-dioxadiazine-3-yl)-4-fluoroisoindole-1,3-dione (II-1a) into a thick-walled, pressure-resistant tube (300 mg, 1.09 mmol, 1 equiv), and dissolve (2-aminoethyl)carbamate tert-butyl ester (226 mg, 1.41 mmol, 1.3 equiv) in 15 mL of anhydrous 1,4-dioxane. Heat and stir at 100 °C for 7 hours. The reaction was checked by TLC until complete. The solvent was removed by vacuum distillation, and the solution was purified by silica gel column chromatography to give a pale yellow solid (II-1b) (199 mg, 0.48 mmol), yield: 41%. MS m / s (ESI) [M+H] + :417.2.

[0092] Step 3: 4-((2-aminoethyl)amino)-2-(2,6-dioxopiperidin-3-yl)isodihydroindole-1,3-dione (II-1c)

[0093] Weigh 190 mg (0.45 mmol, 1 equiv) of tert-butyl (2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)amino)ethyl)carbamate (II-1b) into a round-bottom flask and dissolve in 1 mL of TFA. Stir at room temperature for 1 hour. The reaction was confirmed to be complete by TLC. The trifluoroacetic acid in the solution was concentrated under reduced pressure using an oil pump to obtain a pale yellow crude product (II-1c) (135 mg), which was reserved for the next reaction. Yield: 94%. MS m / s (ESI) [M+H]+ 317.1.

[0094] Step 4: N-(3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)ethyl)-3-(6-((2-methoxyphenyl)sulfonylamino)-1-methyl-2-oxo-1,2-dihydrobenzo[cd]indole-4-yl)propionamide (II-1);

[0095] Weigh 1 g (60 mg, 0.136 mmol, 1 equiv) of 3-(6-((2-methoxyphenyl)sulfonamide)-1-methyl-2-oxo-1,2-dihydrobenzo[cd]indol-4-yl)propionic acid I-1 (60 mg, 0.136 mmol, 1 equiv), HATU (76 mg, 0.20 mmol, 1.5 equiv), and DIPEA (52 mg, 0.40 mmol, 3 equiv) into a round-bottom flask and dissolve in 20 mL of anhydrous DMF. Stir at 0 °C for 20 minutes, then add 4-((2-aminoethyl)amino)-2-(2,6-dioxopiperidin-3-yl)isodihydroindol-1,3-dione (Ⅱ-1c) (56 mg, 0.18 mmol, 1.3 equiv) and stir at room temperature for half an hour. The reaction was completed by TLC, extracted with EA, the organic layer was washed with saturated brine, dried over Mg2SO4, the solvent was removed by vacuum distillation, and purified by silica gel column chromatography to give a pale yellow solid (Ⅱ-1) (74 mg, 0.10 mmol), yield: 74%. MS m / s (ESI) [M+H + : 739.2. 1 H NMR (300MHz, DMSO-d6) δ8.82(s,1H),8.06(s,1H),7.84(s,1H),7.65(s,2H),7.49(t,J=7.9Hz,1H),7.40(t,J=7.7Hz,1H),7.05(d,J=7.6Hz,3 H),6.91(t,J=7.7Hz,1H),6.78(d,J=8.5Hz,1H),6.55(d,J=7.5Hz,1H),6.22(s,2H),4.97(s,1H),4.09(s,3H),3.45(s,2H),3.28(s,3H),3.31 -3.11(m,6H),2.83(dt,J=22.3,10.3Hz,4H),2.58(d,J=7.7Hz,2H).

[0096] Example 3

[0097] N-(3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)propyl)-3-(6-((2-methoxyphenyl)sulfonylamino)-1-methyl-2-oxo-1,2-dihydrobenzo[cd]indole-4-yl)propionamide (II-2)

[0098]

[0099] Step 1: (3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)propyl)tert-butyl carbamate (II-2a)

[0100] The preparation method is similar to step two of Example 2: the preparation of tert-butyl carbamate (II-1b), using 2-(2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)ethyl)carbamate (II-1a) as the raw material, with an input amount of 300 mg (1.09 mmol), and finally obtaining a light yellow solid (II-2a) (182 mg, 0.42 mmol), with a yield of 39%.

[0101] Step 2: 4-((3-aminopropyl)amino)-2-(2,6-dioxopiperidin-3-yl)isodihydroindole-1,3-dione (II-2b).

[0102] Step 3: N-(3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)propyl)-3-(6-((2-methoxyphenyl)sulfonylamino)-1-methyl-2-oxo-1,2-dihydrobenzo[cd]indole-4-yl)propionamide (II-2)

[0103] The preparation method is similar to steps three and four of Example 2. Using (3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)propyl)tert-butyl carbamate (II-2a) as the raw material, with an input amount of 180 mg (0.42 mmol), the preparation was carried out in two steps, ultimately yielding a light yellow solid (II-2) (66 mg, 0.088 mmol), with a yield of 65%. MS m / s (ESI) [M+H] + : 753.2. 1H NMR (300MHz, DMSO-d6) δ8.59(s,1H),8.05(s,1H),7.89(s,1H),7.72(s,2H),7.49(t,J=7.5Hz, 1H),7.43-7.38(m,1H),7.16-6.99(m,3H),6.91(t,J=7.6Hz,1H),6.70(d,J=8.5Hz,1H),6.60( d,J=7.6Hz,1H),5.86(s,2H),4.95(s,1H),4.13(s,3H),3.31(s,3H),3.34-3.31(m,2H),3.18( d,J=7.0Hz,2H),3.08(s,2H),2.89-2.72(m,4H),2.59(t,J=7.2Hz,2H),2.13(d,J=8.5Hz,2H).

[0104] Example 4

[0105] N-(4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)butyl)-3-(6-((2-methoxyphenyl)sulfonylamino)-1-methyl-2-oxo-1,2-dihydrobenzo[cd]indole-4-yl)propionamide(II-3)

[0106]

[0107] Step 1: (3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)butyl)tert-butyl carbamate (II-3a)

[0108] The preparation method is similar to step two of Example 2: the preparation of tert-butyl carbamate (II-1b), using 2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindo-4-yl)amino)ethyl)carbamate (II-1a) as the raw material, with an input amount of 300 mg (1.09 mmol), and finally obtaining a light yellow solid (II-3a) (248 mg, 0.57 mmol), with a yield of 53%.

[0109] Step 2: 4-((3-aminobutyl)amino)-2-(2,6-dioxopiperidin-3-yl)isodihydroindole-1,3-dione (II-3b)

[0110] Step 3: N-(3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)butyl)-3-(6-((2-methoxyphenyl)sulfonylamino)-1-methyl-2-oxo-1,2-dihydrobenzo[cd]indole-4-yl)propionamide (II-3)

[0111] The preparation method is similar to steps three and four of Example 2. Using (3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)butyl)carbamate tert-butyl ester (II-3a) as the raw material, with an input amount of 240 mg (0.56 mmol), the preparation was carried out in two steps, ultimately yielding a light yellow solid (II-3) (69 mg, 0.092 mmol), with a yield of 68%. MS m / s (ESI) [M+H] + : 767.2. 1 H NMR(300MHz,DMSO-d6)δ8.46(s,1H),8.06(s,1H),7.88(s,1H),7.69-7.55(m ,2H),7.48(q,J=9.0,8.6Hz,2H),7.18-6.99(m,3H),6.91(t,J=7.6Hz,1H),6 .79(d,J=8.5Hz,1H),6.57(d,J=7.5Hz,1H),5.92(s,2H),4.93(s,1H),4.12( s,3H),3.27(s,3H),3.26-3.20(m,2H),3.08(s,2H),2.94-2.67(m,4H),2.13 -2.09(m,2H).

[0112] Example 5

[0113] N-(5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)pentyl)-3-(6-((2-methoxyphenyl)sulfonylamino)-1-methyl-2-oxo-1,2-dihydrobenzo[cd]indole-4-yl)propionamide(II-4)

[0114]

[0115] Step 1: (3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)amino)pentyl)tert-butyl carbamate (II-4a)

[0116] The preparation method is similar to step two of Example 2: the preparation of tert-butyl carbamate (II-1b), using 2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindo-4-yl)amino)ethyl)carbamate (II-1a) as the raw material, with an input amount of 300 mg (1.09 mmol), and finally obtaining a light yellow solid (II-4a) (229 mg, 0.50 mmol), with a yield of 46%.

[0117] Step 2: 4-((3-aminopentyl)amino)-2-(2,6-dioxopiperidin-3-yl)isodihydroindole-1,3-dione (II-4b).

[0118] Step 3: N-(3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)pentyl)-3-(6-((2-methoxyphenyl)sulfonylamino)-1-methyl-2-oxo-1,2-dihydrobenzo[cd]indole-4-yl)propionamide (II-4)

[0119] The preparation method is similar to steps three and four of Example 2. Using (3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)pentyl)carbamate tert-butyl ester (II-4a) as the raw material, with an input amount of 220 mg (0.48 mmol), the preparation was carried out in two steps, ultimately yielding a light yellow solid (II-4) (67 mg, 0.085 mmol), with a yield of 63%. MS m / s (ESI) [M+H] + : 781.3. 1 H NMR (300MHz, DMSO-d6) δ8.36(s,1H),8.05(s,1H),7.89(s,1H),7.66(dd,J=7.8,1.7Hz,2H),7.54-7.41(m,3H),7.14-7.02(m,3H),6.92(t, J=7.6Hz,1H),6.81(d,J=8.5Hz,1H),6.59(d,J=7.5Hz,1H),6.13(s,1H),5.67(s,1H),4.91(d,J=5.4Hz,1H),4.12(s,3H),3.31(s,3H),3.20 -3.11(m,6H),2.87-2.76(m,2H),2.55(t,J=7.3Hz,2H),1.53(t,J=7.3Hz,2H),1.44-1.37(m,2H),1.33-1.16(m,4H).

[0120] Example 6

[0121] N-(6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)hexyl)-3-(6-((2-methoxyphenyl)sulfonylamino)-1-methyl-2-oxo-1,2-dihydrobenzo[cd]indole-4-yl)propionamide (II-5)

[0122]

[0123] Step 1: (3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)amino)hexyl)tert-butyl carbamate (II-5a)

[0124] The preparation method is similar to step two of Example 2: the preparation of tert-butyl carbamate (II-1b), using 2-(2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)ethyl)carbamate (II-1a) as the raw material, with an input amount of 300 mg (1.09 mmol), and finally obtaining a light yellow solid (II-5a) (240 mg, 0.51 mmol), with a yield of 47%.

[0125] Step 2: 4-((3-aminohexyl)amino)-2-(2,6-dioxopiperidin-3-yl)isodihydroindole-1,3-dione (II-5b).

[0126] Step 3: N-(3-((2-(2,6-dioxopiperidin-3-yl)-1,3-d-dioxoisoindole-4-yl)amino)hexyl)-3-(6-((2-methoxyphenyl)sulfonylamino)-1-methyl-2-oxo-1,2-dihydrobenzo[cd]indole-4-yl)propionamide (II-5)

[0127] The preparation method is similar to steps three and four of Example 2. Using (3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)hexyl)tert-butyl carbamate (II-5a) as the raw material, with an input amount of 240 mg (0.50 mmol), the preparation was carried out in two steps, ultimately yielding a light yellow solid (II-5) (63 mg, 0.081 mmol). The yield of the final step was 59%. MS m / s (ESI) [M+H] + : 795.3. 1H NMR (300MHz, DMSO-d6) δ8.36(s,1H),8.05(s,1H),7.89(s,1H),7.66(dd,J=7.8,1.7Hz,2H),7.54-7.41(m,3H),7.14-7.02(m,3H),6.92(t, J=7.6Hz,1H),6.81(d,J=8.5Hz,1H),6.59(d,J=7.5Hz,1H),6.13(s,1H),5.67(s,1H),4.91(d,J=5.4Hz,1H),4.12(s,3H),3.31(s,3H),3.20 -3.11(m,6H),2.87-2.76(m,2H),2.55(t,J=7.3Hz,2H),1.53(t,J=7.3Hz,2H),1.44-1.37(m,2H),1.33-1.16(m,4H).

[0128] Example 7

[0129] N-(4-(3-(4-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)ethyl)piperazin-1-yl)-3-oxopropyl)-1-methyl-2-oxo-1,2-dihydrobenzo[cd]indole-6-yl)-2-methoxybenzenesulfonamide (II-6)

[0130]

[0131] Step 1: 4-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)amino)ethyl)piperazine-1-carbamate tert-butyl ester (II-6a)

[0132] The preparation method is similar to step two of Example 2: the preparation of tert-butyl carbamate (II-1b), using 2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)ethyl)carbamate (II-1a) as the raw material, with an input amount of 300 mg (1.09 mmol), and finally obtaining a light yellow solid (II-6a) (80 mg, 0.165 mmol), with a yield of 15%.

[0133] Step 2: 2-(2,6-dioxopiperidin-3-yl)-4-((2-(piperazin-1-yl)ethyl)amino)isoindole-1,3-dione (II-6b).

[0134] Step 3: N-(4-(3-(4-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)ethyl)piperazin-1-yl)-3-oxopropyl)-1-methyl-2-oxo-1,2-dihydrobenzo[cd]indole-6-yl)-2-methoxybenzenesulfonamide (II-6)

[0135] The preparation method is similar to steps three and four of Example 2. Using 4-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)ethyl)piperazine-1-carbamate tert-butyl ester (II-6a) as the starting material, it was prepared in two steps, ultimately yielding a light yellow solid (II-6) (49 mg, 0.062 mmol). The yield of the final step was 45%. MS m / s (ESI) [M+H] + : 808.3. 1 H NMR(300MHz,Chloroform-d)δ8.36(s,1H),7.97(s,1H),7.83(s,1H),7.60(dd,J=7.7,1.8Hz ,1H),7.42(p,J=3.9Hz,3H),7.01(dd,J=7.9,5.2Hz,3H),6.89-6.75(m,2H),6.63(s,1H),6. 54(d,J=7.5Hz,1H),4.91-4.80(m,1H),4.04(s,3H),3.69-3.53(m,3H),3.50-3.38(m,3H),3 .26(s,5H),3.14(dt,J=7.0,3.5Hz,2H),2.77(s,2H),2.68-2.59(m,4H),2.49-2.32(m,4H).

[0136] Example 8

[0137] N-(2-(2-((2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)ethoxy)ethyl)-3-(6-(((2-methoxyphenyl)sulfonylamino)-1-methyl-2-oxo-1,2-dihydrobenzo[cd]indole-4-yl)propionamide (II-7)

[0138]

[0139] Step 1: 2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-4-yl)amino)ethoxy)tert-butyl ethyl formate (II-7a)

[0140] The preparation method is similar to step two of Example 2: the preparation of tert-butyl carbamate (II-1b), using 2-(2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)ethyl)carbamate (II-1a) as the raw material, with an input amount of 300 mg (1.09 mmol), and finally obtaining a light yellow solid (II-7a) (105 mg, 0.229 mmol), yield: 21%.

[0141] Step 2: 4-((2-(2-aminoethoxy)ethyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione (II-7b).

[0142] Step 3: N-(2-(2-((2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)ethoxy)ethyl)-3-(6-(((2-methoxyphenyl)sulfonylamino)-1-methyl-2-oxo-1,2-dihydrobenzo[cd]indole-4-yl)propionamide (II-7)

[0143] The preparation method is similar to steps three and four of Example 2. Using 4-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)amino)ethyl)piperazine-1-carbamate tert-butyl ester (II-7a) as the starting material, it was prepared in two steps, ultimately yielding a light yellow solid (II-7) (56 mg, 0.073 mmol). The yield of the final step was 53%. MS m / s (ESI) [M+H] + : 783.2. 1H NMR(400MHz,Chloroform-d)δ9.07(s,1H),8.10(s,0H),7.97(d,J=3.5Hz,2H),7.84(d,J=3.6Hz,1H),7.70(dd,J=8.0,3.4Hz, 1H),7.46(td,J=8.1,7.5,4.6Hz,2H),7.20-7.14(m,1H),7.10-6.99(m,2H),6.90(td,J=7.7,3.5Hz,1H),6.77(dd,J=8.6,3.7 Hz,1H),6.56(dd,J=7.7,3.6Hz,1H),6.47(s,1H),6.29(s,1H),4.93-4.87(m,1H),4.05(s,3H),3.65-3.52(m,4H),3.51-3.36 (m,3H),3.30(s,3H)3.15(dd,J=7.5,3.7Hz,2H),2.83-2.63(m,3H),2.58-2.51(m,2H),2.12-2.02(m,2H),1.95-1.85(s,2H).

[0144] The above description is merely a preferred embodiment of the present invention, and the scope of protection of the present invention is not limited thereto. Any simple changes or equivalent substitutions of the technical solutions that can be obviously obtained by those skilled in the art within the scope of the technology disclosed in the present invention shall fall within the scope of protection of the present invention.

Claims

1. A benzindole bifunctional molecule derivative characterized in that: the derivative is N-(3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)propyl)-3-(6-((2-methoxyphenyl)sulfonylamino)-1-methyl-2-oxo-1,2-dihydrobenzo[cd]indol-4-yl)propanamide, N-(4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)butyl)-3-(6-((2-methoxyphenyl)sulfonylamino)-1-methyl-2-oxo-1,2-dihydrobenzo[cd]indol-4-yl)propanamide, or N-(5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)pentyl)-3-(6-((2-methoxyphenyl)sulfonylamino)-1-methyl-2-oxo-1,2-dihydrobenzo[cd]indol-4-yl)propanamide.

2. Use of the derivative of the bifunctional molecule of the benzindole class according to claim 1 for the preparation of a medicament for the prevention or treatment of a clinical condition associated with BET proteins. The disease associated with BET proteins is rheumatoid arthritis, gouty arthritis, melanoma, liver cancer, kidney cancer, acute leukemia, multiple myeloma, lymphoma, non-small cell lung cancer, prostate cancer, thyroid cancer, skin cancer, pancreatic cancer, ovarian cancer, breast cancer, myelodysplastic syndrome, esophageal cancer, gastrointestinal cancer or mesothelioma.

3. Use according to claim 2, characterized in that, ​

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