Multi-target difunctional chimera compound as well as preparation method and application thereof

Abstract

The invention provides a multi-target difunctional chimera compound as well as a preparation method and application thereof, and belongs to the technical field of biological medicines. On the basis of NEO2734 molecules, a dual-function multi-target PROTAC molecule is designed and synthesized, BET, CBP / p300 and transcription factors (such as tumor progression driving protein AR and AR variant (ARV) in advanced prostate cancer) interacting with the BET, the CBP / p300 and the proteins can be effectively degraded in a targeted manner, so that growth and survival of tumors are inhibited, and the tumor progression driving protein AR and AR variant (ARV) can be inhibited. Especially, under a low dosage, the compound can play a catalytic role and degrade BET and CBP / p300 proteins, BET or CBP / p300 interaction transcription factors which cannot be degraded by NEO2734 micromolecules, and the like, and has stronger in-vivo and in-vitro anti-tumor activity and smaller in-vivo toxic and side effects.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to a multi-target bifunctional chimera compound and a preparation method and application thereof. Background Art

[0002] Epigenetics refers to a heritable regulatory mode that can affect gene expression without involving changes in DNA sequence (genetic mutations and variations). It is a genetic or transmission mechanism that can be intervened and reversed. Studies have shown that the occurrence and development of tumors are related to abnormal gene expression and the activity of gene expression products, and epigenetic changes are one of the key factors leading to abnormal tumor gene expression and the activity of gene expression products. Epigenetic-related modification enzymes (DNA methyltransferases, histone methyltransferases, demethylases, acetyltransferases and deacetylases, etc.) change the structure of chromosomes or recruit specific effector proteins to regulate gene expression through methylation modification of genomic DNA molecules and methylation, demethylation, acetylation and deacetylation of chromatin histones.

[0003] The level of histone acetylation is a dynamic balance controlled by "writers" and "erasers" with opposite enzymatic activities, while the bromodomain and extra-terminal domain (BET) family proteins are an important class of "readers" of epigenetic marks. Cyclic AMP response element binding protein (CBP) and E1A binding protein (p300) with a molecular weight of 300 kilodaltons are very important histone and non-histone acetyltransferases. These epigenetic factors interact with transcription factors, especially tissue- or tumor-specific transcription factors, to form powerful super enhancers, thereby achieving sustainable expression of tissue- or tumor-specific genes.

[0004] Compared with normal cells, tumor cells have characteristics such as unrestricted proliferation, highly active metabolism, and escape from apoptosis, and the maintenance of these phenotypes requires the activation of tumor-specific super enhancers and the high expression of tumor driver genes, thereby promoting the malignant progression of tumors. Therefore, tumor super enhancers should be effective targets for specifically targeting tumors.

[0005] NEO2734 is a dual-target inhibitor of BET and CBP / p300, and its clinical anti-tumor activity is still being tested in various solid tumors and hematological tumors. Summary of the invention

[0006] The purpose of the present invention is to provide a multi-target bifunctional chimeric compound and its preparation method and application. The chimeric compound of the present invention can significantly downregulate BET bromodomain family proteins such as BRD4, CBP / p300 and related tumor-driving transcription factors and other proteins, and has significant anti-tumor activity.

[0007] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0008] The present invention provides a multi-target bifunctional chimeric compound, the structure of which includes a target recognition-binding part, a linking part and a ubiquitin ligase recognition-binding part;

[0009] The target recognition-binding portion includes the following structure:

[0010]

[0011] Preferably, the link part has the following structure:

[0012]

[0013] Wherein, X and Z are independently selected from -S-, -SO-, -SO 2 -、-CH 2 -, -(C=O)-, -NH-, -O-, alkynyl, or the following structure, or does not exist;

[0014]

[0015] Among them, R 1 and R 2 Independently selected from hydrogen or an alkane or cycloalkane of 1 to 6 carbons;

[0016] U, W, and Y are independently selected from saturated or unsaturated straight-chain hydrocarbon groups of 1 to 14 carbon atoms, oxa-chains, phenyl groups, heterocyclic groups, or any of the following structures, wherein the heterocyclic group is piperazinyl, pyrrolyl, pyrazolyl, furanyl, thienyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridinyl, pyrimidinyl, pyrazinyl, or pyridazinyl; or are absent;

[0017]

[0018] Among them, R 3 and R 4 Independently selected from hydrogen or alkanes or cycloalkanes with 1 to 6 carbon atoms; n and m are independently selected from integers of 0 to 10; when n and m are 0, it means that the group does not exist; D and E are independently selected from O, -NH- or -CONH-.

[0019] Preferably, the ubiquitin ligase recognition-binding portion includes but is not limited to compounds that can recognize and bind to the following ubiquitin ligases: Cereblon (CRBN), von Hippel-Lindau (VHL), Murine double minute 2 (MDM2), Inhibitor of apoptosis proteins (IAPs), Aryl hydrocarbon receptor (AhR), Fem-1 homolog B (FEM1B), Kelch-like ECH-associated protein 1 (KEAP1), Ringfinger protein 4 (RNF4), Lethal (3) malignant brain tumor-like protein 3 (L3MBTL3), DDB1-cullin 4-associated factor 1 (DCAF1), DCAF11, DCAF15 or DCAF16; the compounds include but are not limited to the following structures:

[0020]

[0021] The present invention also provides a multi-target bifunctional chimera compound, wherein the compound contains any of the following structures:

[0022]

[0023]

[0024]

[0025] The present invention also provides optical isomers of the above-mentioned multi-target bifunctional chimeric compound, or solvates thereof, or pharmaceutically acceptable salts thereof, or prodrugs thereof, or tautomers thereof, or mesoforms thereof, or racemates thereof, or enantiomers thereof, or diastereomers thereof, or mixtures thereof, or metabolites thereof, or metabolic precursors thereof, or isotope-substituted forms thereof.

[0026] The present invention also provides a method for preparing the above-mentioned multi-target bifunctional chimera compound, comprising the following steps:

[0027]

[0028] The specific reaction steps are as follows: m1 and m2 undergo substitution reaction to generate m3, m3 undergoes reduction to obtain intermediate m4, m4 and m5 undergo condensation to generate m6; m7 undergoes substitution to obtain m8, m8 undergoes coupling to generate m9; m6 and m9 undergo coupling to obtain m10, m10 undergoes deprotection, and then undergoes amide condensation with an acid to obtain the compound of the present invention.

[0029]

[0030] The specific reaction steps are as follows: m1 and m12 undergo substitution reaction to generate m13, m13 is reduced to obtain intermediate m14, m14 and m15 are condensed to generate m16; m16 and m9 are coupled to obtain m17, m17 is deprotected, and then undergoes amide condensation with an acid to obtain the compound of the present invention.

[0031] The present invention also provides the use of the multi-target bifunctional chimera compound or the multi-target bifunctional chimera compound prepared by the above preparation method in the preparation of anti-tumor drugs.

[0032] Preferably, the tumor is a solid tumor or a hematological tumor.

[0033] The present invention also provides the use of the above-mentioned multi-target bifunctional chimeric compound, or the multi-target bifunctional chimeric compound prepared by the above-mentioned preparation method in the preparation of a drug for alleviating or treating cancer, characterized in that the cancer includes but is not limited to acute monocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia mixed lineage leukemia, NUT midline carcinoma, multiple myeloma, glioma, lung cancer, neuroblastoma, Burkitt's lymphoma, cervical cancer, esophageal cancer, nasopharyngeal cancer, ovarian cancer, pancreatic cancer, colorectal cancer, prostate cancer, kidney cancer, bladder cancer, urothelial carcinoma, adrenal cancer, breast cancer or any one of sarcomas originating from bone and soft tissue.

[0034] The present invention also provides an anti-tumor drug, which contains the multi-target bifunctional chimera compound or the multi-target bifunctional chimera compound prepared by the above preparation method.

[0035] Beneficial effects of the present invention:

[0036] Based on the NEO2734 molecule, the present invention designs and synthesizes a dual-functional multi-target PROTAC molecule, which can effectively target and degrade BET, CBP / p300 and transcription factors that interact with these proteins (such as AR and AR variants (ARV) proteins, which are tumor progression drivers in advanced prostate cancer), thereby inhibiting tumor growth and survival.

[0037] Compared with the original occupancy-driven small molecule inhibitor NEO2734 targeting BET and CBP / p300 proteins, the compounds of the present invention have the following advantages:

[0038] 1) Catalyzes and degrades BET and CBP / p300 proteins at low doses;

[0039] 2) It can degrade transcription factors (interacting with BET and / or CBP / p300 proteins) that NEO2734 cannot target;

[0040] 3) Stronger anti-tumor activity in vitro and in vivo; BRIEF DESCRIPTION OF THE DRAWINGS

[0041] Figure 1 The degradation activity of compounds 19D-F (Examples 24-26) and C-005 (Example 5) of the present invention on BRD4, CBP / P300, AR and AR variants;

[0042] Figure 2 The inhibitory activity of compounds 19D-F (Examples 24-26) of the present invention on prostate cancer colony formation;

[0043] Figure 3 and Figure 4 The killing effect of the compounds 19D-F (Examples 24-26) and C-005 (Example 5) of the present invention on prostate cancer cells;

[0044] Figure 5 The inhibitory activity of compounds 19D-F (Examples 24-26) of the present invention on tumor growth and their effects on mouse body weight in a SCID mouse subcutaneous transplant tumor model. DETAILED DESCRIPTION

[0045] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0046] Example 1

[0047] The synthetic route of the compound of the present invention is as follows:

[0048]

[0049] The structural formula of the synthesized compound is as follows:

[0050]

[0051]

[0052]

[0053] Among them, the specific preparation methods corresponding to each compound are as follows:

[0054] Step 1: Synthesis of 5-bromo-1,3-dimethylpyridin-2(1H)-one:

[0055] 5-Bromo-3-methylpyridin-2(1H)-one (10 g, 53 mmol) was dissolved in DMF (50 mL), iodomethane (6.62 mL, 106 mmol) and potassium carbonate (22 g, 160 mmol) were added, and the mixture was reacted at room temperature for 12 hours. The reaction solution was poured into water, extracted, and the organic phases were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, and column chromatography was performed to obtain 9.24 g of the product (yield 86%). LC / MS (ESI+) calculated for C 7 H 9 BrNO([M+H] + )m / z 202.0, found 202.2.

[0056] Step 2: Synthesis of 1,3-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaboropolysaccharide-2-yl)pyridin-2(1H)-one:

[0057] 5-Bromo-1,3-dimethylpyridin-2(1H)-one (3.6 g, 18 mmol) was dissolved in 1,4-dioxane (25 mL), and 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborane) (2.3 g, 27 mmol), potassium acetate (5.2 g, 54 mmol), palladium chloride (0.65 g, 0.9 mmol) were added, and nitrogen was charged and heated to 90 degrees for 12 hours. The reaction solution was poured into water, extracted, and the organic phases were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, and column chromatography to obtain 2.48 g of the product (yield 56%), LC / MS (ESI+) calcd for C 13 H 21 BNO 3 ([M+H] + )m / z 250.2, found 250.3.

[0058] Step 3: Synthesis of tert-butyl (2-(2-((5-bromo-2-nitrophenyl)amino)ethoxy)ethyl)carbamate:

[0059] 4-Bromo-2-fluoro-1-nitrobenzene (4.4 g, 20 mmol) was dissolved in DMF (10 mL), tert-butyl (2-(2-aminoethoxy)ethyl) carbamate (4 g, 22 mmol) and triethylamine (6.6 mL, 66 mmol) were added, and the mixture was reacted at room temperature for 12 hours. The reaction solution was poured into water, extracted, and the organic phases were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, and column chromatography was performed to obtain 7.16 g of the product (yield 88%). LC / MS (ESI+) calculated for C 15 H 23 Bn 3 O 5 ([M+H] + )m / z 404.1, found 404.2.

[0060] Step 4: Synthesis of tert-butyl (2-((5-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-nitrophenyl)amino)ethoxy)ethyl)aminobenzoate:

[0061] Tert-butyl (2-(2-((5-bromo-2-nitrophenyl)amino)ethoxy)ethyl)carbamate (2g, 5mmol) was dissolved in 1,4-dioxane (10mL), 1,3-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaboropolylan-2-yl)pyridin-2(1H)-one (1.84g, 7.5mmol), cesium carbonate (3.4g, 10mmol), tetrakis(triphenylphosphine)palladium (0.56g, 0.5mmol) were added, nitrogen was filled for protection, and the mixture was heated to 85 degrees for 12 hours. The reaction solution was poured into water, extracted, the organic phases were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, and column chromatography to obtain 1.85 grams (84% yield) of the product, LC / MS (ESI+) calculated for C 22 H 31 N 4 O 6 ([M+H] + )m / z 447.2, found 447.5.

[0062] Step 5: Synthesis of tert-butyl (2-((2-amino-5-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)phenyl)amino)ethoxy)amino)ethyl)amino)carbamate:

[0063] Tert-butyl (2-((5-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-nitrophenyl)amino)ethoxy)ethyl)aminobenzoate (1.85 g, 4 mmol) was dissolved in methanol (5 mL), palladium carbon (44 mg, 0.4 mmol) was added, hydrogen was reduced, and the reaction was carried out at room temperature for 4 hours. The palladium carbon was removed by filtration through diatomaceous earth, and 1.5 g of the product (yield 83%) was obtained by column chromatography. LC / MS (ESI+) calculated for C 22 H 33 N 4 O 4 ([M+H] + )m / z 417.2, found 417.5.

[0064] Step 6: Synthesis of tert-butyl (2-(2-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazol-1-yl)ethoxy)ethyl)carbamate:

[0065] Dissolve tert-butyl (2-((2-amino-5-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)phenyl)amino)ethoxy)amino)ethyl)amino)carbamate (1.5 g, 3.6 mmol) in methanol (5 mL), add tetrahydro-2H-pyran-4-carbaldehyde (400 μL, 3.8 mmol), acetic acid (1 mL, 18 mmol), and react at room temperature for 6 hours. Add saturated sodium bicarbonate solution to quench, extract, combine the organic phases, wash with saturated brine, dry over anhydrous magnesium sulfate, and obtain 1.32 g of the product by column chromatography (yield 72%), LC / MS (ESI+) calcd for C 28 H 39 N 4 O 5 ([M+H] + )m / z 511.3, found 511.6.

[0066] Step 7: Synthesis of 5-(1-(2-(2-aminoethoxy)ethyl)-2-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazol-6-yl)-1,3-dimethylpyridin-2(1H)-one:

[0067] Tert-butyl (2-(2-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazol-1-yl)ethoxy)ethyl)carbamate (1.32 g, 2.5 mmol) was dissolved in methanol (5 mL), and hydrochloric acid (4 M / L in 1,4-Dioxane, 6.45 mL, 25.8 mmol) was added and reacted at room temperature for 4 hours. The product was dried under reduced pressure to obtain 0.82 g (yield 77%). LC / MS (ESI+) calculated for C 23 H 31 N 4 O 3 ([M+H] + )m / z 411.2, found411.5.

[0068]

[0069] Step 1: Synthesis of 5-bromo-2-nitro-N-(2-(trifluoromethoxy)ethyl)aniline:

[0070] 4-Bromo-2-fluoro-1-nitrobenzene (6 g, 27 mmol) was dissolved in DMF (30 mL), 2-(trifluoromethoxy)ethane-1-amine (5 g, 30 mmol) and N,N-diisopropylethylamine (15.8 mL, 89 mmol) were added, and the mixture was reacted at room temperature for 12 hours. The reaction solution was poured into water, extracted, and the organic phases were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, and column chromatography was performed to obtain 8.7 g of the product (yield 98%). LC / MS (ESI+) calculated for C 9 H 9 F 3 N 2 O 3 ([M+H] + )m / z 329.0, found 329.4.

[0071] Step 2: Synthesis of 1,3-dimethyl-5-(4-nitro-3-)((2-(trifluoromethoxy)ethyl)amino)phenyl)pyridin-2(1H)-one:

[0072] 5-Bromo-2-nitro-N-(2-(trifluoromethoxy)ethyl)aniline (6 g, 18 mmol) was dissolved in 1,4-dioxane (30 mL), and 1,3-dimethyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaboropolylan-2-yl)pyridin-2(1H)-one (6.8 g, 27 mmol), cesium carbonate (12 g, 36 mmol), tetrakis(triphenylphosphine)palladium (2.1 g, 1.8 mmol) were added, nitrogen was filled, and the mixture was heated to 85 degrees for 12 hours. The reaction solution was poured into water, extracted, and the organic phases were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, and column chromatography to obtain 5.5 g of the product (yield 81%), LC / MS (ESI+) calcd for C 16 H 17 F 3 N 3 O 4 ([M+H] + )m / z 372.1, found 372.3.

[0073] Step 3: Synthesis of 5-(4-amino-3-((2-(trifluoromethoxy)ethyl)amino)phenyl)-1,3-dimethylpyridin-2(1H)-one:

[0074] 1,3-Dimethyl-5-(4-nitro-3-)((2-(trifluoromethoxy)ethyl)amino)phenyl)pyridin-2(1H)-one (5 g, 13 mmol) was dissolved in methanol (15 mL), palladium carbon (139 mg, 1.3 mmol) was added, hydrogen was reduced, and the reaction was carried out at room temperature for 4 hours. The palladium carbon was removed by filtration through diatomaceous earth, and 3.6 g of the product (yield 78%) was obtained by column chromatography. LC / MS (ESI+) calculated for C 16 H 19 F 3 N 3 O 2 ([M+H] + )m / z 342.1424, found 342.1415.

[0075] Step 4: Synthesis of tert-butyl 4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidine-1-carboxylate:

[0076] 5-(4-amino-3-((2-(trifluoromethoxy)ethyl)amino)phenyl)-1,3-dimethylpyridin-2(1H)-one (3 g, 8.7 mmol) was dissolved in methanol (10 mL), 4-formylpiperidine-1-carboxylic acid tert-butyl ester (1.9 g, 9 mmol) and acetic acid (2.5 mL, 44 mmol) were added, and the mixture was reacted at room temperature for 10 hours. The reaction solution was poured into water, extracted, and the organic phases were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, and column chromatography was performed to obtain 3.3 g of the product (yield 70%). LC / MS (ESI+) calcd for C 27 H 34 F 3 N 4 O 4 ([M+H] + )m / z 535.2527, found 535.2511.

[0077] Step 5: Synthesis of 1,3-dimethyl-5-(2-(piperidin-4-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-6-yl)pyridin-2(1H)-one:

[0078] Dissolve tert-butyl 4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidine-1-carboxylate (3 g, 5.6 mmol) in methanol (5 mL), add hydrochloric acid (4 M / L in 1,4-Dioxane, 5.6 mL, 56 mmol), and react at room temperature for 4 hours. The product was dried under reduced pressure to give 1.8 g (yield 74%). LC / MS (ESI+) calculated for C 22 H 26 F 3 N 4 O 2 ([M+H] + )m / z 435.2, found 435.4.

[0079] Example 1: 4-((4-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)-4-oxobutyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0080]

[0081] 1,3-Dimethyl-5-(2-(piperidin-4-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-6-yl)pyridin-2(1H)-one (50 mg, 0.1 mmol), 4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)butyric acid (38 mg, 0.1 mmol), HATU (48 mg, 0.12 mmol) and DIPEA (40 μL, 2.2 mmol) were added to 3 mL of DMF in sequence and reacted at room temperature for 12 hours. The reaction solution was poured into water, extracted, the organic phases were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, and column chromatography to obtain 56 mg of the product (yield 65%), LC / MS (ESI+) calculated for C 37 H 37 F 3 N 7 O 7 ([M+H] + )m / z 748.3, found 748.7. 1H NMR (400MHz, DMSO-D6) δ11.12(s,1H),7.98(d,J=2.6Hz,1H),7.83–7.76(m,2H),7.66–7.55(m,2H),7.40(dd,J=8.4 ,1.6Hz,1H),7.18–7.12(m,2H),7.08(d,J=7.0Hz,1H),5.08(dd,J=12.9,5.4Hz,1H),4.72(d,J=5.4Hz,2H),4.52(d, J=12.7Hz,1H),4.46(t,J=4.9Hz,2H),4.34–4.18(m,2H),4.14–4.01(m,1H),3.54(s,3H),3.26(d,J=12.3Hz,1H),2 .95–2.83(m,2H),2.65–2.53(m,2H),2.11(s,3H),2.08–2.00(m,1H),1.94(d,J=6.9Hz,2H),1.76(d,J=11.2Hz,1H).

[0082] Example 2: 4-((6-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)-6-oxooxy)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0083]

[0084] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 39 H 41 F 3 N 7 O 7 ([M+H]+)m / z 776.3014, found 776.3065.

[0085] Example 3: 4-((8-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)-8-oxyoctyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0086]

[0087] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 41 H 45 F 3 N 7 O 7 ([M+H] + )m / z 804.3, found 804.8.

[0088] Example 4: 4-((10-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)-10-oxodecanyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0089]

[0090] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 43 H 49 F 3 N 7 O 7 ([M+H] + )m / z 832.3640, found 832.3609.

[0091] Example 5: 4-((12-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)-12-oxododecanyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0092]

[0093] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 45 H 53 F 3 N 7 O 7 ([M+H] + )m / z 860.4, found 860.9.

[0094] Example 6: 4-((2-(2-(2-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)-2-oxoethoxy)ethoxy)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0095]

[0096] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 47 H 57 F 3 N 7 O 7 ([M+H] + )m / z 888.4, found 888.9.

[0097] Example 7: 4-((2-(2-(2-(2-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)-2-oxoethoxy)ethoxy)ethyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0098]

[0099] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C41 H 45 F 3 N 7 O 9 ([M+H] + )m / z 836.3225, found 836.3297.

[0100] Example 8: N-(2-(2-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazol-1-yl)ethoxy)ethyl)-2-((2-(2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)acetamide)acetamide

[0101]

[0102] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 43 H 49 F 3 N 7 O 10 ([M+H] + )m / z 880.3, found 880.9.

[0103] Example 9: N-(2-(2-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazol-1-yl)ethoxy)ethyl)-4-((2-(2-(2-6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)butanamide

[0104]

[0105] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 38 H 42 N 7 O 8 ([M+H] + )m / z 724.3, found 724.8.

[0106] Example 10: N-(2-(2-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazol-1-yl)ethoxy)ethyl)-6-((2-(2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)hexanamide

[0107]

[0108] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 40 H 46 N 7 O 8 ([M+H] + )m / z 752.3402, found 752.3485.

[0109] Example 11: N-(2-(2-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazol-1-yl)ethoxy)ethyl)-8-((2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)octanamide

[0110]

[0111] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 42 H 50 N 7 O 8 ([M+H] + )m / z 780.4, found 780.9.

[0112] Example 12: N-(2-(2-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazol-1-yl)ethoxy)ethyl)-10-((2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)decanamide

[0113]

[0114] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 44 H 54N 7 O 8 ([M+H] + )m / z 808.4028, found 808.4010.

[0115] Example 13: N-(2-(2-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazol-1-yl)ethoxy)ethyl)-12-((2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)dodecanoamide

[0116]

[0117] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 46 H 58 N 7 O 8 ([M+H] + )m / z 836.4341, found 836.4323.

[0118] Example 14: N-(2-(2-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazol-1-yl)ethoxy)ethyl)-12-((2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)amino)dodecanoamide

[0119]

[0120] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 48 H 62 N 7 O 8 ([M+H] + )m / z 864.5, found 864.4.

[0121] Example 15: N-(2-(2-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazol-1-yl)ethoxy)ethyl)-2-(2-((2-(2-(2-6-dioxopiperidin-3-yl)-1,3-dioxoindolin-4-yl)amino)ethoxy)ethoxy)acetamide

[0122]

[0123] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 42 H 50 N 7 O 10 ([M+H] + )m / z 812.3614, found 812.3685.

[0124] Example 16: N-(2-(2-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazol-1-yl)ethoxy)ethyl)-2-(2-(2-(2-((2-(2-(2-(2-(2-)dioxypiperidin-3-yl)-1,3-dihydroisoindolin-4-yl)amino)ethoxy)ethoxy)ethoxy)acetamide

[0125]

[0126] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 44 H 54 N 7 O 11 ([M+H] + )m / z 856.4, found 856.9.

[0127] Example 17: 5-((2-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)-2-oxoethyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0128]

[0129] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 37 H 37 F 3 N 7 O 7 ([M+H] +)m / z 748.3, found 748.7. 1H NMR (400MHz, DMSO-D6) δ11.04(s,1H),7.95(d,J=2.6Hz,1H),7.77(t,J=3.8Hz,2H),7.55 (dd,J=8.4,3.0Hz,2H),7.38(d,J=8.1Hz,1H),7.13–6.97(m,3H),5.01(dd,J=12.9,5.4H z,1H),4.70(d,J=5.6Hz,2H),4.53–4.38(m,3H),4.25–4.02(m,3H),3.50(s,3H),3.20(m ,3H),2.90–2.77(m,2H),2.60–2.49(m,2H),2.07(s,3H),2.00–1.92(m,2H),1.89(s,2H).

[0130] Example 18: 5-((4-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)-4-oxobutyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0131]

[0132] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 39 H 41 F 3 N 7 O 7 ([M+H] + )m / z 776.3014, found 776.3065. 1 HNMR (400 MHz, DMSO-D 6)δ11.07(s,1H),7.98(d,J=2.7Hz,1H),7.87–7.73(m,2H),7.58(dd,J=8.4,1.5Hz,2H),7.40(dd,J=8.7,2.6Hz,1H),7.21(t,J=5. 4Hz,1H),6.98(d,J=2.1Hz,1H),6.88(dd,J=8.4,2.1Hz,1H),5.03(dd,J=12.9,5.4Hz,1H),4.71(d,J=5.7Hz,2H),4.54(d,J=13.0H z,1H),4.45(t,J=4.9Hz,2H),4.02(d,J=13.3Hz,1H),3.54(s,3H),3.26–3.16(m,3H),2.87(ddd,J=17.4,14.1,5.5Hz,1H),2.78–2 .68(m,1H),2.56(dd,J=15.0,2.8Hz,2H),2.11(s,3H),2.04–1.94(m,2H),1.85(dt,J=14.5,7.5Hz,6H),1.69(m,1H),1.46(m,1H).

[0133] Example 19: 5-((6-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)-6-oxooxy)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0134]

[0135] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 41 H 45 F 3 N 7 O 7 ([M+H] + )m / z 804.3, found 804.8. 1 H NMR (400 MHz, DMSO-D 6)δ11.07(s,1H),7.97(d,J=2.6Hz,1H),7.83–7.74(m,2H),7.56(dd,J=8.4,3.3Hz,2H),7.38(dd,J=8.4,1.7Hz,1H),7.14(t,J=5.3Hz,1 H),6.95(d,J=2.1Hz,1H),6.85(dd,J=8.3,2.1Hz,1H),5.03(dd,J=12.9,5.4Hz,1H),4.70(t,J=5.1Hz,2H),4.51(d,J=13.0Hz,1H),4.44 (t,J=4.9Hz,2H),4.07–3.95(m,1H),3.53(s,3H),3.17(q,J=7.3Hz,3H),2.87(ddd,J=17.9,14.1,5.4Hz,1H),2.71(t,J=12.6Hz,1H),2 .56(dd,J=19.0,5.3Hz,2H),2.38(q,J=6.9Hz,2H),2.11(s,3H),2.03–1.74(m,5H),1.60(dp,J=14.5,9.2Hz,5H),1.42(q,J=7.0Hz,2H).

[0136] Example 20: N-(2-(2-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazol-1-yl)ethoxy)ethyl)-6-((2-(2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)amino)hexanamide

[0137]

[0138] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 42 H 50 N 7 O 8 ([M+H] + )m / z 780.3715, found 780.3706.

[0139] Example 21: (4R)-1-((S)-2-(5-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)-5-oxopentanamide)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

[0140]

[0141] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 49 H 60 F 3 N 8 O 7 S([M+H] + )m / z 961.4, found 961.1.

[0142] Example 22: (4R)-1-((S)-2-(6-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)-6-oxohexanaphthamide)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

[0143]

[0144] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 50 H 62 F 3 N 8 O 7 S([M+H] + )m / z 975.4, found 975.1.

[0145] Example 23: (4R)-1-((S)-2-(7-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)-7-oxothiazolamide)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

[0146]

[0147] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 51 H 64 F 3 N 8 O 7 S([M+H] + )m / z 989.5, found 989.1.

[0148] Example 24: (4R)-1-((S)-2-(8-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)-8-oxooctylamide)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

[0149]

[0150] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 52 H 66 F 3 N 8 O 7 S([M+H] + )m / z 1003.4722, found 1003.4671.

[0151] Example 25: (4R)-1-((S)-2-(9-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)-9-oxonamidamide)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

[0152]

[0153] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 53 H 68 F 3 N 8 O 7 S([M+H] + )m / z 1017.4878, found 1017.4832.

[0154] Example 26: (4R)-1-((S)-2-(10-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)-10-oxobutanamide)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

[0155]

[0156] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 54 H 70 F 3 N 8 O 7 S([M+H] + )m / z 1031.5035, found 1031.4982. 1 HNMR(400MHz,CHLOROFORM-D)δ8.65(d,J=2.9Hz,1H),7.74(dd,J=8.3,2.9Hz,1H),7.58–7.4 6(m,2H),7.43–7.30(m,7H),6.29(dd,J=16.8,8.9Hz,1H),4.73(dd,J=14.2,7.7Hz,2H),4.56 (p,J=7.9Hz,6H),4.43–4.28(m,3H),4.18–3.99(m,3H),3.65(s,5H),3.16(d,J=16.8Hz,2H) ,2.69(m,1H),2.50(s,3H),2.36(m,2H),2.26–2.18(m,5H),1.97–1.77(m,10H),1.62(s,4H).

[0157] Example 27: N1-(2-(2-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazol-1-yl)ethoxy)ethyl)-N5-((2S)-1-((4R)-4-hydroxy-2-((4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)glutaramide

[0158]

[0159] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C50 H 65 N 8 O 8 S([M+H] + )m / z 937.5, found 937.7.

[0160] Example 28: N1-(2-(2-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazol-1-yl)ethoxy)ethyl)-N6-((2S)-1-((4R)-4-hydroxy-2-((4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)hexanamide

[0161]

[0162] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 51 H 67 N 8 O 8 S([M+H] + )m / z 951.5, found 951.7.

[0163] Example 29: N1-(2-(2-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazol-1-yl)ethoxy)ethyl)-N7-((2S)-1-((4R)-4-hydroxy-2-((4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)heptanediamide

[0164]

[0165] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 52 H 69 N 8 O 8 S([M+H] + )m / z 965.5, found 965.6.

[0166] Example 30: N1-(2-(2-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazol-1-yl)ethoxy)ethyl)-N8-((2S)-1-((4R)-4-hydroxy-2-((4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)octanamide

[0167]

[0168] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 53 H 71 N 8 O 8 S([M+H] + )m / z 979.5, found 979.7.

[0169] Example 31: N1-(2-(2-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazol-1-yl)ethoxy)ethyl)-N9-((2S)-1-((4R)-4-hydroxy-2-((4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)nonanamide

[0170]

[0171] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 54 H 73 N 8 O 8 S([M+H] + )m / z 993.5, found 993.2.

[0172] Example 32: N1-(2-(2-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-2-(tetrahydro-2H-pyran-4-yl)-1H-benzo[d]imidazol-1-yl)ethoxy)ethyl)-N10-((2S)-1-((4R)-4-hydroxy-2-((4-methylthiazol-5-yl)benzyl)carbamoyl)pyrrolidin-1-yl)-3,3-dimethyl-1-oxobutan-2-yl)decanamide

[0173]

[0174] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 55 H 75 N 8 O 8 S([M+H] + )m / z 1007.5, found 1007.3.

[0175] Example 33: (4R)-1-((S)-2-(12-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)-12-oxododecanamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

[0176]

[0177] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 56 H 74 F 3 N 8 O 7 S([M+H] + )m / z 1059.5, found 1059.6.

[0178] Example 34: (4R)-1-((S)-2-(11-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)-11-oxoundecylamino)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

[0179]

[0180] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 55 H 72 F 3 N 8 O 7 S([M+H] + )m / z 1045.5, found 1045.2.

[0181] Example 35: (4R)-1-((S)-2-(10-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)-10-oxodecanamido)-3,3-dimethylbutanoyl)-4-hydroxy-N-(S)-1-(4-(4-methylthiazol-5-yl)phenyl)ethyl)pyrrolidine-2-carboxamide

[0182]

[0183] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 55 H 72 F 3 N 8 O 7 S([M+H] + )m / z 1045.5, found 1045.2.

[0184] Example 36: (4R)-1-((S)-2-(10-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)decanamide)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

[0185]

[0186] Step 1: Synthesis of tert-butyl 10-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)decanoate:

[0187] 1,3-Dimethyl-5-(2-(piperidin-4-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-6-yl)pyridin-2(1H)-one (80 mg, 0.18 mmol) was dissolved in acetonitrile (5 mL), 10-bromodiol acid tert-butyl ester (85 mg, 0.27 mmol) and potassium carbonate (128 mg, 0.9 mmol) were added, and the mixture was heated to 50 degrees for 12 hours. The reaction solution was poured into water, extracted, and the organic phases were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, and column chromatography to obtain 61 mg of the product (yield 50%), LC / MS (ESI+) calculated for C 36 H 52 F 3 N4 O 4 ([M+H] + )m / z 661.4, found 661.1.

[0188] Step 2: Synthesis of 10-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)decanoic acid:

[0189] Tert-butyl 10-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)decanoate (50 mg) was dissolved in dichloromethane (5 mL) and trifluoroacetic acid was added. The reaction was carried out at room temperature for 4 hours. The mixture was concentrated under reduced pressure to obtain 27 mg of the product (yield 61%). LC / MS (ESI+) calculated for C 32 H 44 F 3 N 4 O 4 ([M+H] + )m / z 605.3, found 605.7.

[0190] Step 3: (4R)-1-((S)-2-(10-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)decanamide)-3,3-dimethylbutanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide

[0191] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 54 H 72 F 3 N 8 O 6 S([M+H] + )m / z 1017.5, found 1017.8.

[0192] Example 37: 4-((3-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)-3-oxopropyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0193]

[0194] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 38 H 39 F 3 N 7 O 7 ([M+H] + )m / z 762.3, found 762.6.

[0195] Example 38: 4-((5-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)-5-oxopentyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0196]

[0197] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 40 H 43 F 3 N 7 O7([M+H] + )m / z 790.3, found 790.8.

[0198] Example 39: 4-((7-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)-7-oxoheptyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0199]

[0200] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 42 H 47 F 3 N 7 O 7 ([M+H] + )m / z 818.3, found 818.8.

[0201] Example 40: 4-((9-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)-9-oxo)amino)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione

[0202]

[0203] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 44 H 51 F 3 N 7 O 7 ([M+H] + )m / z 846.4, found 846.9.

[0204] Example 41: 5-(3-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)-3-oxopropyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione

[0205]

[0206] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 39 H 44 N 7 O 8 ([M+H] + )m / z 762.3, found 762.5.

[0207] Example 42: 5-(5-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)-5-oxopentyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione

[0208]

[0209] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 41 H 48 N 7 O8 ([M+H] + )m / z 790.3, found 790.3.

[0210] Example 43: 5-(7-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)-7-oxoheptyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione

[0211]

[0212] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 43 H 52 N 7 O 8 ([M+H] + )m / z 818.3, found 818.3.

[0213] Example 44: 5-(8-(4-(6-(1,5-dimethyl-6-oxo-1,6-dihydropyridin-3-yl)-1-(2-(trifluoromethoxy)ethyl)-1H-benzo[d]imidazol-2-yl)piperidin-1-yl)-8-oxooctyl)amino)-2-(2,6-dioxopiperidin-3-yl)isoindole-1,3-dione

[0214]

[0215] The target compound was obtained by a preparation method similar to that of Example 1. LC / MS (ESI+) calculation for C 45 H 56 N 7 O 8 ([M+H] + )m / z 832.4, found 832.4.

[0216] Experimental example

[0217] The performance of the compounds synthesized in the examples was tested, and the experimental method was as follows:

[0218] Experimental Example 1: Degradation activity of the compounds of the present invention on BRD4, CBP / p300, AR and AR variant (ARV)

[0219] Experimental methods:

[0220] After the prostate cancer cell line 22Rv1 was treated with the control or the indicated concentrations and durations of small molecule compounds, the cells were collected and lysed in RIPA lysis buffer (Thermo Scientific) with the addition of protease inhibitors (MCE, China). The total protein concentration was determined using a BCA protein assay kit (Beyotime, China). The total protein was transferred to a NC membrane (Millipore) after being processed through a 10% SDS-PAGE gel. After blocking with 5% skim milk for 1 hour at room temperature, the primary antibody was incubated overnight at 4°C. The next day, the NC membrane was incubated in a specific HRP-conjugated secondary antibody for one hour at room temperature. All NC membranes were visualized using an ECL substrate kit (Millipore) and images were acquired using a Bio Spectrum 600 Imaging System (UVP). The antibodies used included anti-p300 (#86377, Cell Signaling Technology), CBP (#7389, Cell Signaling Technology), BRD4 (#13440, Cell Signaling Technology), AR (#5153, Cell Signaling Technology), HRP-conjugated goat anti-rabbit IgG (H+L) (AS014, ABclonal), and HRP-conjugated goat anti-mouse IgG (H+L) (AS003, ABclonal).

[0221] Experimental Example 2: Inhibitory activity of the compounds of the present invention on prostate cancer colony formation

[0222] Experimental methods:

[0223] Prostate cancer cell lines (as shown) were cultured at 1×10 3 The cells were inoculated in 6-well plates at a density of 100 cells and cultured for 14 days. The cell colonies were fixed with 4% polyformaldehyde (Beyotime, China) for 30 minutes at room temperature and stained with 0.1% crystal violet (SigmaAldrich) for 30 minutes at room temperature. The cell colonies with more than 50 cells were counted.

[0224] Experimental Example 3: Inhibitory activity of the compounds of the present invention on prostate cancer cells

[0225] Experimental methods:

[0226] Cell viability was detected using CCK-8 assay (Promega, USA) according to the instructions. 3Cell lines (as shown) were plated in 96-well plates, with 100 μL of culture medium per well, and treated with the indicated concentration of drugs for the indicated time. CCK-8 solution (10 μL) was added to each well and incubated at 37°C for 1 hour. The absorbance at 450 nm was detected using an automatic microplate reader (Synergy4; BioTek., Winooski, VT, USA). Background fluorescence was subtracted, and the signal was normalized to DMSO-treated cells. The dose-response curves were fitted and statistically analyzed using the nonlinear regression function in GraphPad Prism 8.0 software.

[0227] Experimental Example 4: Antitumor activity of the compounds of the present invention in a SCID mouse subcutaneous transplant tumor model.

[0228] 22Rv1 cells (3×10 6 ) was mixed with matrigel (50 μl 1×PBS + 50 μl matrigel (BD Biosciences)) and injected subcutaneously into 6-week-old SCID male mice. 3 After the size of the mice was measured, the animals were randomly divided into 5 treatment groups (n=6 / group), including control (10% DMSO and 90% corn oil), NEO2734 (10 mg / kg), 19D (10 mg / kg), 19E (10 mg / kg) or 19F (10 mg / kg). The treatment method was intraperitoneal injection every day for 21 consecutive days. At the end of the experiment, the mice were killed and the tumors were removed.

[0229] The results are as follows Figures 1 to 4 As shown:

[0230] according to Figure 1 It can be seen that the compounds 19F (Example 26) and C005 (Example 5) of the present invention can effectively degrade BRD4, CBP / p300, AR and AR variant (ARV).

[0231] according to Figure 2 It can be seen that compounds 19E (Example 25) and 19F (Example 26) can effectively inhibit the colony formation of prostate cancer cells 22Rv1 and DU145 at 0.03 μmol, and their activity is better than that of the control compound NEO2734.

[0232] according to Figure 3 and Figure 4 It can be seen that the inhibitory activity of the compounds of the present invention on prostate cancer cells 22Rv1 and DU145 is at the nanomolar level, and its activity is significantly better than that of the control compound NEO2734.

[0233] according to Figure 5It can be seen that at the same dosage, compared with the control compound NEO2734, the compound 19D-F of the present invention has stronger growth inhibitory activity on mouse transplanted tumors, and the compound of the present invention has no obvious effect on the body weight of mice, which proves that the toxicity is low.

[0234] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A multi-target bifunctional chimera compound, characterized in that: The structure of the bifunctional chimeric compound includes a target recognition-binding portion, a linker portion, and a ubiquitin ligase recognition-binding portion; The target recognition-binding portion includes the following structure:

2. The multi-target bifunctional chimera compound according to claim 1, characterized in that: The link part has the following structure: Wherein, X, Z are independently selected from -S-, -SO-, -SO2-, -CH2-, -(C=O)-, -NH-, -O-, alkynyl, or the following structures, or do not exist; Wherein, R1 and R2 are independently selected from hydrogen or an alkane or cycloalkane of 1 to 6 carbons; U, W, and Y are independently selected from saturated or unsaturated straight-chain hydrocarbon groups of 1 to 14 carbon atoms, oxa-chains, phenyl groups, heterocyclic groups, or any of the following structures, wherein the heterocyclic group is piperazinyl, pyrrolyl, pyrazolyl, furanyl, thienyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyridinyl, pyrimidinyl, pyrazinyl, or pyridazinyl; or are absent; Wherein, R3 and R4 are independently selected from hydrogen or alkanes or cycloalkanes with 1 to 6 carbon atoms; n and m are independently selected from integers of 0 to 10; when n, m is 0, it means that the group does not exist; D and E are independently selected from O, -NH- or -CONH-.

3. The multi-target bifunctional chimera compound according to claim 1, characterized in that: The ubiquitin ligase recognition-binding portion includes but is not limited to compounds that can recognize and bind to the following ubiquitin ligases: Cereblon, von Hippel-Lindau, Murine double minute 2, Inhibitor of apoptosis proteins, Aryl hydrocarbon receptor, Fem-1 homolog B, Kelch-like ECH-associated protein 1, Ringfinger protein 4, Lethal 3 malignant brain tumor-like protein 3, DDB1-cullin 4-associated factor 1, DCAF11, DCAF15 or DCAF16; the compounds include but are not limited to the following structures:

4. A multi-target bifunctional chimera compound, characterized in that: The compound contains any of the following structures:

5. An optical isomer of the multi-target bifunctional chimeric compound according to any one of claims 1 to 4, or a solvate thereof, or a pharmaceutically acceptable salt thereof, or a prodrug thereof, or a tautomer thereof, or a mesoform thereof, or a racemate thereof, or an enantiomer thereof, or a diastereomer thereof, or a mixture thereof, or a metabolite thereof, or a metabolic precursor thereof, or an isotope-substituted form thereof.

6. Use of the multi-target bifunctional chimeric compound according to any one of claims 1 to 4, or the multi-target bifunctional chimeric compound prepared by the preparation method according to claim 5, in the preparation of anti-tumor drugs.

7. The use according to claim 6, characterized in that: The tumor is a solid tumor or a blood tumor.

8. Use of the multi-target bifunctional chimeric compound according to any one of claims 1 to 4, or the multi-target bifunctional chimeric compound prepared by the preparation method according to claim 5, in the preparation of a drug for alleviating or treating cancer, characterized in that: The cancer includes, but is not limited to, acute monocytic leukemia, acute myeloid leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia, mixed lineage leukemia, NUT midline carcinoma, multiple myeloma, glioma, lung cancer, neuroblastoma, Burkitt's lymphoma, cervical cancer, esophageal cancer, nasopharyngeal cancer, ovarian cancer, pancreatic cancer, colorectal cancer, prostate cancer, kidney cancer, bladder cancer, urothelial carcinoma, adrenal cancer, breast cancer, or any one of sarcomas originating from bone and soft tissue.

9. An anti-tumor drug, characterized in that: The drug contains the multi-target bifunctional chimeric compound according to any one of claims 1 to 4, or the multi-target bifunctional chimeric compound prepared by the preparation method according to claim 5.

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