A chimera molecule and its preparation method and use

Abstract

The present invention provides a chimeric molecule, a preparation method and use thereof, and relates to the pharmaceutical field. The chimeric molecule is a compound composed of a unit M, a unit N and a connecting unit L, or a salt thereof, or a stereoisomer thereof, or a solvate thereof, wherein both ends of the connecting unit L are connected to an atom on the unit M and the unit N, respectively. The compound of the present invention, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a solvate thereof can induce degradation of the BRD4 protein, thereby exerting a therapeutic effect on tumors or other diseases that are dependent on BRD4. Such compounds are expected to be used as one of the components of a pharmaceutical preparation for the treatment of tumors or other diseases that are dependent on BRD4.

Description

Technical Field

[0001] The present invention relates to the pharmaceutical field, and in particular to a chimera molecule and a preparation method and use thereof. Background Art

[0002] Malignant tumors are one of the most serious public health problems in the world. In 2012, there were approximately 14.1 million new cases of malignant tumors and approximately 8.2 million deaths worldwide. Chemotherapy has always played an important role in the treatment of tumors due to its wide adaptability and thorough elimination of tumor cells. Among them, traditional chemotherapy drugs represented by cytotoxic drugs have been the basis of tumor treatment in the past few decades, but have gradually faded out of the historical stage of tumor treatment due to their serious toxic side effects. Then, tumor targeted drugs represented by kinase inhibitors have revolutionized the treatment of malignant tumors in the past 10 years due to their advantages such as rapid efficacy and mild toxic side effects. However, these drugs can only control tumor growth in a relatively short period of time due to the rapid generation of drug-resistant mutations. Epigenetic target drugs that have emerged in recent years have the advantages of long-lasting efficacy and slow development of drug resistance, which make up for the shortcomings of traditional kinase inhibitors and bring a new revolution to the treatment of malignant tumors.

[0003] Bromodomain containing protein 4 (BRD4) is a member of the BET (bromodomain and extraterminal) family. This protein can recognize acetylated lysine sites on histones, and then recruit transcription factors to the vicinity of promoters and enhancers to initiate gene expression. BRD4 small molecule inhibitors inhibit the expression of tumor-related genes by inhibiting the connection between BRD4 and acetylated histones, thereby achieving the effect of treating tumors. Current studies have shown that BRD4 inhibitors exhibit broad activity and persistent inhibitory effects in lymphomas, acute leukemias, and various solid tumors. BRD4 inhibitors exert their anti-tumor effects in hematological malignancies such as acute myeloid leukemia, multiple myeloma, chronic lymphocytic leukemia, and B-cell acute lymphocytic leukemia by reducing the expression of the proto-oncogene c-MYC and causing cell cycle arrest.

[0004] Small molecule protein degradation inducers are a class of drugs that, after binding to target proteins, can induce ubiquitination of target proteins in a variety of ways and ultimately be degraded by proteasomes or lysosomes. There are many types of protein degradation inducing molecules, among which the most mature are the proteolysis-targeting chimeras (PROTACs) developed by CMCrews and JEBradner et al. Such molecules generally contain three structural units: ① target protein ligand molecule fragments, which are used to provide affinity between PROTAC molecules and the proteins to be degraded; ② E3 ubiquitin ligase ligands, which are used to provide affinity between PROTAC molecules and E3 ubiquitin ligases, so that specific E3 ubiquitin ligases can approach the proteins to be degraded under the action of PROTAC molecules, induce ubiquitination of target proteins and ultimately degrade them; ③ connection fragments, which are used to connect the target protein ligand and the E3 ubiquitin ligase ligand, to ensure that the PROTAC molecule can interact with the target protein and the E3 ubiquitin ligase at the same time.

[0005] At present, a large number of literatures have reported PROTAC molecules developed for a variety of disease-related proteins. Compared with ordinary small molecule inhibitors, PROTAC molecules have the following advantages: ① The drug works by degrading proteins rather than inhibiting protein function, so the drug effect is more durable and the activity is better; ② One molecule of compound can often induce the degradation of multi-molecule proteins, so the target protein can be fully degraded at substoichiometric drug concentrations, the effective dose of the drug is lower, and the safety may be better; ③ The drug molecule does not need to bind to the functional domain of the target protein to exert protein degradation, so PROTAC molecules can be developed into drugs for proteins that could not be drug targets before, such as cytoskeletal proteins or transcription factors; ④ The interaction between E3 ubiquitin ligase and target protein can make PROTAC molecules more selective than traditional small molecule drugs, so it can avoid the safety problems caused by the off-target effects of traditional drugs to a certain extent. These advantages of PROTAC molecules make it particularly suitable for development as anti-tumor drugs. Its feature of exerting drug efficacy by degrading rather than inhibiting the function of target proteins can be used to design drugs to overcome drug-resistant mutations that do not affect the affinity of drugs to target proteins. Many studies have shown that in terms of anti-cancer activity, degrading proteins is better than inhibiting proteins. Summary of the invention

[0006] The present invention provides a class of compounds, or stereoisomers thereof, or pharmaceutically acceptable salts thereof, or solvates thereof, which can induce degradation of BRD4 protein, thereby exerting good anti-tumor activity on a variety of BRD4-dependent tumors, and can therefore be used to treat related tumors. Since BRD4 is also related to the occurrence and development of diabetes, atherosclerosis, coronary artery disease, etc., the compounds provided by the present invention can also be used to treat these diseases. In addition, since the BRD4 protein degradation molecules combined with other drugs show good synergistic anti-tumor activity against a variety of tumors, the compounds provided by the present invention can also be used as one of the effective ingredients of a combination of drugs for the treatment of different tumors.

[0007] The present invention is achieved in that:

[0008] The present invention provides a compound represented by formula (I), or a salt thereof, or a stereoisomer thereof, or a solvate thereof:

[0009] MLN

[0010] (I)

[0011] Among them, L is a connecting unit, and both ends of L are connected to an atom on unit M and unit N respectively;

[0012] The unit M is shown in formula (a):

[0013]

[0014] in,

[0015] R1 is selected from substituted or unsubstituted -C 1-6 Alkyl, -C 3-6 Cycloalkyl, -C 5-6 cycloalkenyl, substituted or unsubstituted aryl or heteroaryl;

[0016] The substituents of the alkyl group in R1 are independently selected from -C 3-6 Cycloalkyl, -C 5-6 cycloalkenyl, substituted or unsubstituted aryl or heteroaryl;

[0017] R2 is selected from hydrogen or -C 1-6 alkyl;

[0018] R3 is a substituent on the benzene ring, independently selected from hydrogen, halogen, -C 1-3 Haloalkyl, -C 1-6 Alkyl, -C 3-6 Cycloalkyl, -OC 1-6 Alkyl, -OC 3-6 Cycloalkyl, -NHC 1-6 Alkyl, -NHC 3-6 Cycloalkyl, -C(O)NR9R 10、-CH2NR 11 R 12 、-NHC(O)R 13 ;

[0019] R4 is selected from hydrogen, -C 1-6 Alkyl, -C 3-6 Cycloalkyl;

[0020] R9 and R 10 are independently selected from hydrogen, -C 1-6 Alkyl, -C 3-6 Cycloalkyl, -C 4-7 Heterocycloalkyl, or R9 and R 10 It forms a saturated ring containing multiple heteroatoms with the N atom;

[0021] R 11 With R 12 are independently selected from hydrogen, -C 1-6 Alkyl, -C 3-6 Cycloalkyl, -C 4-7 Heterocycloalkyl, or R 11 and R 12 It forms a saturated ring containing multiple heteroatoms with the N atom;

[0022] R 13 independently selected from hydrogen, -C 1-6 Alkyl, -C 3-6 Cycloalkyl, -C 4-7 Heterocycloalkyl,

[0023] n1 is selected from 0, 1, 2, 3, 4;

[0024] Unit N is shown in formula (b):

[0025]

[0026] in,

[0027] R6 is a substituent on the benzene ring, independently selected from hydrogen, -C 1-6 Alkyl, -C 1-3 Haloalkyl, halogen, cyano, C 2-6 Alkynyl, C 2-6 Alkenyl, hydroxyl, amino, -OC 1-6 Alkyl, cyclopropyloxy;

[0028] n2 is selected from 0, 1, 2, 3;

[0029] X is selected from -CR 14 R 15 -;

[0030] R 14 With R 15are independently selected from hydrogen, halogen, hydroxyl, -C 1-6 Alkyl; or R 14 With R 15 Formation = O;

[0031] L is selected from -Ak2-Z-Ak1-Y-;

[0032] Y is selected from -O-, -NR 16 - or None;

[0033] Z is selected from -O-, -NR 17 or none;

[0034] R 16 With R 17 are independently selected from hydrogen, -C 1-6 alkyl;

[0035] Ak1 is selected from -C(O)(CH2) n3 -、-(CH2) n4 C(O)-、-(CH2) n5 -、-(CH2CH2O) n10 CH2CH2- or none;

[0036] Ak2 is selected from -C(O)(CH2) n6 -、-(CH2) n7 -、-(CH2CH2O) n8 -、-(CH2CH2O) n9 CH2CH2- or none;

[0037] n3, n4, n5, n6, and n7 are each independently selected from an integer between 0 and 15;

[0038] n8, n9, and n10 are each independently selected from an integer between 0 and 5;

[0039] When R1 is selected from C5 cycloalkyl, R2 is not hydrogen.

[0040] Further,

[0041] In unit M,

[0042] R2 is selected from hydrogen or methyl;

[0043] In unit N,

[0044] R6 are independently selected from H, F, Cl, Br, -CN, -C≡CH, -CH=CH2, -CF3, -CHF2, -CH2F, -OH, -NH2, methoxy, ethoxy, cyclopropyloxy;

[0045] X is selected from -CH2-, -C(O)-, -CF2-, -CHF-, -CH(OH)-, -CH(CH3)-.

[0046] Further,

[0047] The unit M is shown in formula (c):

[0048]

[0049] in,

[0050] R1 is selected from substituted or unsubstituted -C 1-6 Alkyl, -C 3-6 Cycloalkyl, -C 5-6 cycloalkenyl, substituted or unsubstituted aryl or heteroaryl;

[0051] The substituents of the alkyl group in R1 are independently selected from -C 3-6 Cycloalkyl, -C 5-6 cycloalkenyl, substituted or unsubstituted aryl or heteroaryl;

[0052] R2 is selected from hydrogen or -C 1-6 alkyl;

[0053] R3 is a substituent on the benzene ring, independently selected from hydrogen, halogen, -C 1-3 Haloalkyl, -C 1-6 Alkyl, -C 3-6 Cycloalkyl, -OC 1-6 Alkyl, -OC 3-6 Cycloalkyl, -NHC 1-6 Alkyl, -NHC 3-6 Cycloalkyl;

[0054] R4 is selected from hydrogen, -C 1-6 Alkyl, -C 3-6 Cycloalkyl;

[0055] n1 is selected from 0, 1, 2;

[0056] A is selected from wherein A1 and A2 are independently selected from -C(O)NH-, -NHC(O)-, -C(O)-, -CH2-, -NH-, -O- or none, and Cy1 is selected from or none;

[0057] Wherein B1 and B2 are each independently selected from N or CH, m1 and m2 are each independently selected from 0, 1, 2;

[0058] Preferably,

[0059] The unit M is shown in formula (d):

[0060]

[0061] in,

[0062] R1 is selected from substituted or unsubstituted -C 1-6 Alkyl, -C 3-6 Cycloalkyl, -C 5-6 cycloalkenyl, substituted or unsubstituted aryl or heteroaryl;

[0063] The substituents of the alkyl group in R1 are independently selected from -C 3-6 Cycloalkyl, -C 5-6 cycloalkenyl, substituted or unsubstituted aryl or heteroaryl;

[0064] R2 is selected from hydrogen or -C 1-6 alkyl;

[0065] R3 is selected from hydrogen, halogen, -C 1-3 Haloalkyl, -C 1-6 Alkyl, -C 3-6 Cycloalkyl, -OC 1-6 Alkyl, -OC 3-6 Cycloalkyl;

[0066] R4 is selected from H, methyl, ethyl, propyl, isopropyl, cyclopropyl, cyclobutyl;

[0067] A is selected from wherein A1 and A2 are independently selected from -C(O)NH-, -NHC(O)-, -C(O)-, -CH2-, -NH-, -O- or none, and Cy1 is selected from or none;

[0068] Wherein B1 and B2 are each independently selected from N or CH, and m1 and m2 are each independently selected from 0, 1, or 2.

[0069] Further,

[0070] The unit M is shown in formula (e):

[0071]

[0072] in,

[0073] R1 is selected from -C 1-6 Alkyl, -C 3-6 Cycloalkyl or substituted methyl;

[0074] The substituent of the methyl group in R1 is selected from -C 3-6 Cycloalkyl, -C 5-6 cycloalkenyl, substituted or unsubstituted aryl or heteroaryl;

[0075] R2 is selected from hydrogen or -C 1-6alkyl;

[0076] R3 is selected from hydrogen, halogen, -C 1-3 Haloalkyl, -C 1-6 Alkyl, -C 3-6 Cycloalkyl, -OC 1-6 Alkyl, -OC 3-6 Cycloalkyl;

[0077] R4 is selected from H, methyl, ethyl, propyl, isopropyl, cyclopropyl, cyclobutyl;

[0078] A is selected from wherein A1 and A2 are independently selected from -C(O)NH-, -NHC(O)-, -C(O)-, -CH2-, -NH-, -O- or none, and Cy1 is selected from or none;

[0079] Wherein B1 and B2 are each independently selected from N or CH, and m1 and m2 are each independently selected from 0, 1, or 2.

[0080] Further,

[0081] R1 is selected from -C 1-6 Alkyl, -C 3-6 Cycloalkyl or substituted methyl;

[0082] The substituent of the methyl group in R1 is selected from -C 3-6 Cycloalkyl, -C 5-6 Cycloalkenyl, phenyl, thienyl, furanyl, pyrrolyl, pyridinyl, pyrimidinyl, imidazole;

[0083] R2 is selected from hydrogen or methyl;

[0084] R3 is selected from hydrogen, halogen, -OC 1-6 Alkyl, -OC 3-6 Cycloalkyl;

[0085] R4 is selected from H, methyl, ethyl, propyl, isopropyl, cyclopropyl, cyclobutyl;

[0086] A is selected from

[0087] Preferably,

[0088] R1 is selected from -C 1-3 Alkyl, -C 3-6 Cycloalkyl or substituted methyl;

[0089] The substituent of the methyl group in R1 is selected from -C 3-6 Cycloalkyl, -C 5-6Cycloalkenyl, phenyl, thienyl, furanyl, pyrrolyl, pyridinyl, pyrimidinyl, imidazole;

[0090] R2 is selected from methyl;

[0091] R3 is selected from hydrogen, -OC 1-6 alkyl;

[0092] R4 is selected from H, methyl, ethyl;

[0093] A is selected from

[0094] Further,

[0095] Unit N is shown in formula (f):

[0096]

[0097] in,

[0098] X is selected from -CR 14 R 15 -;

[0099] R 14 With R 15 are independently selected from hydrogen, halogen, hydroxyl, -C 1-6 Alkyl; or R 14 With R 15 Formation = O;

[0100] Preferably,

[0101] X is selected from -CH2-, -C(O)-.

[0102] Further,

[0103] L is selected from -Ak2-Z-Ak1-Y-;

[0104] Y is selected from -O-, -NH- or none;

[0105] Z is selected from -O-, -NH- or none;

[0106] Ak1 is selected from -C(O)(CH2) n3 -、-(CH2) n4 C(O)-、-(CH2) n5 -、-(CH2CH2O) n10 CH2CH2- or none;

[0107] Ak2 is selected from -C(O)(CH2) n6 -、-(CH2) n7 -、-(CH2CH2O) n8 -、-(CH2CH2O) n9CH2CH2- or none;

[0108] n3, n4, n5, n6, and n7 are each independently selected from an integer between 0 and 15;

[0109] n8, n9, and n10 are each independently selected from an integer between 0 and 5;

[0110] Preferably,

[0111] L is selected from n is selected from an integer between 0-10.

[0112] Furthermore, the compound is as shown in formula (II):

[0113]

[0114] in,

[0115] R1, R2, R3, R4, A, L and X are selected from the above.

[0116] Furthermore, the compound is represented by formula (III):

[0117]

[0118] in,

[0119] R1, A, L and X are selected from the group consisting of those mentioned above.

[0120] Furthermore, the compound is represented by formula (IV):

[0121]

[0122] in,

[0123] R1, L and X are selected from the aforementioned.

[0124] Furthermore, the compound is represented by formula (V):

[0125]

[0126] in,

[0127] L and X are selected from the aforementioned.

[0128] Furthermore, the compound is one of the following compounds:

[0129]

[0130]

[0131]

[0132] The present invention also provides the use of the aforementioned compound, or a salt thereof, or a stereoisomer thereof, or a solvate thereof in the preparation of a BRD4 protein degrading agent;

[0133] Preferably, the drug is a drug that has a down-regulating effect on BRD4 and / or its downstream protein c-Myc.

[0134] Furthermore, the drug is a drug for treating BRD4-dependent tumors, diabetes, atherosclerosis, and coronary artery disease.

[0135] Furthermore, the tumor is leukemia, lymphoma, liver cancer, breast cancer, colon cancer, rectal cancer, colorectal cancer, lung cancer, or prostate cancer.

[0136] The present invention also provides a pharmaceutical composition, which is a preparation prepared by using the aforementioned compound, or its salt, or its stereoisomer, or its solvate as an active ingredient and adding pharmaceutically acceptable excipients.

[0137] The compounds and derivatives provided in the present invention can be named according to the IUPAC (International Union of Pure and Applied Chemistry) or CAS (Chemical Abstracts Service, Columbus, OH) nomenclature system.

[0138] Definitions of terms used in the present invention: Unless otherwise stated, the initial definitions provided for groups or terms in this document apply to the groups or terms throughout the specification; for terms that are not specifically defined in this document, the meaning that a person skilled in the art can give them should be given based on the disclosure and context.

[0139] "Substitution" refers to the replacement of a hydrogen atom in a molecule by another different atom or molecule.

[0140] The structures of the compounds described in the present invention all refer to structures that can exist stably.

[0141] In the present invention, the minimum and maximum values ​​of the carbon atom content in the hydrocarbon group are indicated by prefixes, for example, the prefix (C a-b )alkyl refers to any alkyl group containing from "a" to "b" carbon atoms; thus, for example, C 1-6 Alkyl refers to a straight or branched chain alkyl group containing 1 to 6 carbon atoms; for example, C 3-6 Cycloalkyl refers to a cycloalkyl group consisting of 3, 4, 5 or 6 carbon atoms; 5-6 The cycloalkenyl group refers to a cycloalkenyl group consisting of 5 or 6 carbon atoms.

[0142] In the present invention, halogen is fluorine, chlorine, bromine or iodine.

[0143] In the present invention, the haloalkyl group is an alkyl group substituted with halogen.

[0144] In the present invention, heterocycloalkyl refers to a cycloalkyl group in which carbon atoms are replaced by one or more heteroatoms.

[0145] In the present invention, the heteroatom is selected from O, S, N and the like.

[0146] In the present invention, when an atom on the unit M and the unit N is connected to L, the hydrogen on the atom is removed and connected to L.

[0147] The beneficial effect of the present invention is that the compound of the present invention, or its stereoisomer, or its pharmaceutically acceptable salt, or its solvate can induce the degradation of BRD4 protein, thereby exerting a therapeutic effect on BRD4-dependent tumors or other diseases. Such compounds are expected to be used as one of the components of pharmaceutical preparations for the treatment of BRD4-dependent tumors or other diseases.

[0148] Obviously, according to the above contents of the present invention, in accordance with common technical knowledge and customary means in the art, without departing from the above basic technical ideas of the present invention, other various forms of modification, replacement or change may be made.

[0149] The following is a further detailed description of the above contents of the present invention through specific implementation methods in the form of embodiments. However, this should not be understood as the scope of the above subject matter of the present invention being limited to the following examples. All technologies implemented based on the above contents of the present invention belong to the scope of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0150] Figure 1 The changes in the protein levels of BRD2 / 3 / 4, PLK1, and c-Myc after the VCaP cells were treated with Examples 3 and 9 for 24 hours.

[0151] Figure 2 The changes in the levels of BRD4 and c-Myc proteins after treating LNCaP, 22Rv1, VCaP and DU145 cells with Example 9 (1 μM) for different time periods.

[0152] Figure 3 The changes in BRD4 and c-Myc protein levels after 22Rv1, VCaP and DU145 cells were treated with different concentrations of Example 9 for 24 hours. DETAILED DESCRIPTION

[0153] The raw materials and equipment used in the specific embodiments of the present invention are all known products and are obtained by purchasing commercially available products.

[0154] Example 1, 4-((R)-5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-N-(1-(5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)pentyl)piperidin-4-yl)-3-methoxybenzamide

[0155]

[0156] Step 1: Synthesis of (R)-4-((5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-3-methoxy-N-(piperidin-4-yl)benzamide (1a)

[0157] (R)-7-chloro-5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin (3.40 g, 10 mmol) was dissolved in 1,4-dioxane (40 mL), and tert-butyl 4-(4-amino-3-methoxybenzamide)piperidine-1-carboxylate (3.50 g, 10 mmol), Pd2(dba)3 (915 mg, 1 mmol), Xphos (953 mg, 2 mmol) and Cs2CO3 (6.50 g, 20 mmol) were added to the solution. The mixture was degassed with nitrogen and stirred at 110°C for 12 h until the starting material was completely converted. The mixture was then extracted with EA / H2O, and the organic phase was purified by flash column chromatography (PE / EA=2 / 1, v / v) to give tert-butyl (R)-4-(4-((5-(cyclohexylmethyl))-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-3-methoxybenzamidopiperidine-1-carboxylate (3.08 g, yield: 46.3%). The intermediate was then dissolved in DCM / TFA (V / V=2 / 1, 30 mL) and the mixture was stirred at room temperature for 6 hours. After the reaction was completed, the mixture was concentrated under reduced pressure to give a light yellow solid product 1a (2.19 g, 3.91 mmol, yield 84%). ESI-MS [M+H]+ (M / z): 560.4.

[0158] Step 2: Synthesis of target product

[0159] Intermediate 1a (128 mg, 0.19 mmol) and 4-((5-bromopentyl)oxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (0.284 mmol) were dissolved in DMF (3 mL), and NaHCO3 (31 mg, 0.387 mmol) was added to the above solution. The above mixture was stirred at 65 ° C until the starting material was completely converted. The reaction mixture was quenched with 20 mL of water and extracted with 3×15 mL of EA. The combined organic layers were washed with 30 mL of saturated NaHCO3 and dried over MgSO4. After filtration and concentration under reduced pressure, the product was purified by flash column chromatography (PE / EA=3 / 1, v / v) to give a white solid (target product). (68.4 mg, 0.076 mmol, yield 40%) 1 H NMR (400MHz, CDCl3) δ8.47(d,J=8.4Hz,1H),8.30(s,1H),7.80(s,1H),7.68(dd,J=8.5,7.3Hz,1H),7.45(dd,J=4.5,2.8Hz,2H ),7.30(dd,J=8.5,1.8Hz,1H),7.21(d,J=8.5Hz,1H),6.34(d,J=8.1Hz,1H),4.97(dd,J=7.8,2.8Hz,1H),4.88(dd,J=7.9,3.9 Hz,1H),4.31(dd,J=13.7,6.4Hz,1H),4.19(m,2H),3.98(s,3H),3.02(s,2H),2.87–2.72(m,8H),2.60–2.51(m,2H),2.36–2.2 6(m,2H),2.15–2.04(m,3H),1.89–1.59(m,14H),1.39–1.11(m,6H),1.03(d,J=16.5Hz,2H),0.86(t,J=7.4Hz,3H).HRMS(ESI) + :calcd for C 48 H 60 N 11 O7[M+H] + :902.4672,found:902.4670

[0160] Example 2, 4-((R)-5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-N-(1-(6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)hexyl)piperidin-4-yl)-3-methoxybenzamide

[0161]

[0162] Intermediate 1a (128 mg, 0.19 mmol) and 4-((6-bromohexyl)oxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (0.284 mmol) were dissolved in DMF (3 mL), and NaHCO3 (31 mg, 0.387 mmol) was added to the above solution. The above mixture was stirred at 65 ° C until the starting material was completely converted. The reaction mixture was quenched with 20 mL of water and extracted with 3×15 mL of EA. The combined organic layers were washed with 30 mL of saturated NaHCO3 and dried over MgSO4. After filtration and concentration under reduced pressure, the product was purified by flash column chromatography (PE / EA=3 / 1, v / v) to give a white solid (target product). (57.7 mg, 0.063 mmol, yield 33%) 1 H NMR (400MHz, CDCl3) δ8.48(d,J=8.4Hz,1H),8.30(s,1H),7.80(s,1H),7.68(dd,J=8.5,7.3Hz,1H),7.49–7.43(m,2H),7.30(dd,J=8.5,1 .9Hz,1H),7.22(d,J=8.4Hz,1H),6.31(d,J=8.1Hz,1H),4.97(dd,J=11.8,5.3Hz,1H),4.88(dd,J=7.9,4.0Hz,1H),4.31(dd,J=13.7,6.3 Hz,1H),4.23–4.15(m,3H),3.99(s,3H),3.16(d,J=11.5Hz,2H),2.91–2.73(m,8H),2.61(q,J=7.8Hz,2H),2.43(q,J=11.4Hz,2H),2.20– 2.10(m,2H),1.94–1.66(m,14H),1.57(m,3H),1.44(m,2H),1.22(d,J=29.1Hz,3H),1.09–0.96(m,2H),0.87(t,J=7.4Hz,3H).HRMS(ESI) + :calcd for C 49 H 62 N 11 O7[M+H] + :916.4828,found:916.4832.

[0163] Example 3, 4-((R)-5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-N-(1-(7-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)heptyl)piperidin-4-yl)-3-methoxybenzamide

[0164]

[0165] Intermediate 1a (128 mg, 0.19 mmol) and 4-((7-bromoheptyl)oxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (0.284 mmol) were dissolved in DMF (3 mL), and NaHCO3 (31 mg, 0.387 mmol) was added to the above solution. The above mixture was stirred at 65 ° C until the starting material was completely converted. The reaction mixture was quenched with 20 mL of water and extracted with 3×15 mL of EA. The combined organic layers were washed with 30 mL of saturated NaHCO3 and dried over MgSO4. After filtration and concentration under reduced pressure, the product was purified by flash column chromatography (PE / EA=3 / 1, v / v) to give a white solid (target product). (72.2 mg, 0.078 mmol, yield 41%)1H NMR (400MHz, CDCl3) δ8.47(d,J=8.4Hz,1H),8.29(s,1H),7.78(s,1H),7.67(dd,J=8.5,7.3Hz,1H),7.47–7.42(m,2H),7.29(d,J=1.9Hz ,1H),7.21(d,J=8.4Hz,1H),6.18(d,J=8.0Hz,1H),4.99–4.86(m,2H),4.31(dd,J=13.7,6.3Hz,1H),4.26–4.14(m,2H),3.99(s,3H),3. 02(d,J=10.7Hz,2H),2.92–2.65(m,8H),2.43(q,J=7.4Hz,2H),2.23(t,J=11.8Hz,2H),2.12–2.02(m,3H),1.92–1.63(m,12H),1.55(m, 4H),1.43(q,J=6.3,5.5Hz,2H),1.35(q,J=7.6,6.1Hz,2H),1.27–1.14(m,4H),1.01(t,J=13.1Hz,2H),0.87(t,J=7.4Hz,3H).HRMS(ESI) + :calcd for C 50 H 64 N 11O7[M+H] + :930.4985,found:930.4986.

[0166] Example 4, ((R)-5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-N-(1-(8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)octyl)piperidin-4-yl)-3-methoxybenzamide

[0167]

[0168] Intermediate 1a (128 mg, 0.19 mmol) and 4-((8-bromooctyl)oxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (0.284 mmol) were dissolved in DMF (3 mL), and NaHCO3 (31 mg, 0.387 mmol) was added to the above solution. The above mixture was stirred at 65 ° C until the starting material was completely converted. The reaction mixture was quenched with 20 mL of water and extracted with 3×15 mL of EA. The combined organic layers were washed with 30 mL of saturated NaHCO3 and dried over MgSO4. After filtration and concentration under reduced pressure, the product was purified by flash column chromatography (PE / EA=3 / 1, v / v) to give a white solid (target product). (84.2 mg, 0.089 mmol, yield 47%) 1 H NMR (400MHz, CDCl3) δ8.47(d,J=8.4Hz,1H),8.30(s,1H),7.81(s,1H),7.66(s,1H),7.44(dd,J=4.7,2.8Hz,2H),7.26(d,J=1.9Hz, 1H),7.21(d,J=8.5Hz,1H),6.16(d,J=7.9Hz,1H),4.98–4.91(m,1H),4.88(dd,J=7.9,4.0Hz,1H),4.31(dd,J=13.7,6.3Hz,1H),4. 17(m,2H),4.00–3.95(m,3H),2.99(s,2H),2.88–2.68(m,8H),2.38(t,J=7.8Hz,2H),2.20–2.10(m,3H),2.07–2.01(m,2H),1.87(m ,6H),1.69(dd,J=25.0,8.5Hz,6H),1.51(q,J=7.3Hz,4H),1.39–1.18(m,10H),1.06–0.97(m,2H),0.86(t,J=7.4Hz,3H).HRMS(ESI)+ :calcd for C 51 H 66 N 11 O7[M+H] + :944.5141,found:944.5145.

[0169] Example 5, 4-((R)-5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-N-(1-(9-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)nonyl)piperidin-4-yl)-3-methoxybenzamide

[0170]

[0171] Intermediate 1a (128 mg, 0.19 mmol) and 4-((9-bromononyl)oxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (0.284 mmol) were dissolved in DMF (3 mL), and NaHCO3 (31 mg, 0.387 mmol) was added to the above solution. The above mixture was stirred at 65 ° C until the starting material was completely converted. The reaction mixture was quenched with 20 mL of water and extracted with 3×15 mL of EA. The combined organic layers were washed with 30 mL of saturated NaHCO3 and dried over MgSO4. After filtration and concentration under reduced pressure, the product was purified by flash column chromatography (PE / EA=3 / 1, v / v) to give a white solid (target product). (67.2 mg, 0.070 mmol, yield 37%) 1H NMR (400MHz, CDCl3) δ8.47(dd,J=8.4,2.4Hz,1H),8.30(s,1H),7.79(d,J=2.4Hz,1H),7.70–7.62(m,1H),7.44(dd,J=6.4,2.2 Hz,2H),7.26(d,J=1.9Hz,1H),7.20(d,J=8.5Hz,1H),6.10(d,J=10.2Hz,1H),4.98–4.85(m,2H),4.35–4.27(m,1H),4.23–4.1 2(m,2H),3.98(d,J=2.8Hz,3H),3.03(s,2H),2.91–2.63(m,8H),2.38(dd,J=20.2,11.7Hz,2H),2.22–2.11(m,3H),2.09–2.01 (m,2H),1.92–1.64(m,12H),1.58–1.49(m,4H),1.36–1.16(m,12H),1.08–0.96(m,2H),0.86(m,J=7.4,1.7Hz,3H).HRMS(ESI) + :calcdfor C 52 H 68 N 11 O7[M+H] + :958.5298,found:958.5303.

[0172] Example 6, 4-((R)-5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-N-(1-(10-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)decyl)piperidin-4-yl)-3-methoxybenzamide

[0173]

[0174] Intermediate 1a (128 mg, 0.19 mmol) and 4-((10-bromodecanyl)oxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (0.284 mmol) were dissolved in DMF (3 mL), and NaHCO3 (31 mg, 0.387 mmol) was added to the above solution. The above mixture was stirred at 65 ° C until the starting material was completely converted. The reaction mixture was quenched with 20 mL of water and extracted with 3×15 mL of EA. The combined organic layers were washed with 30 mL of saturated NaHCO3 and dried over MgSO4. After filtration and concentration under reduced pressure, the product was purified by flash column chromatography (PE / EA=3 / 1, v / v) to give a white solid (target product). (70 mg, 0.072 mmol, yield 38%) 1 H NMR (400MHz, CDCl3) δ8.48(d,J=8.4Hz,1H),8.29(s,1H),7.78(s,1H),7.66(dd,J=8.5,7.3Hz,1H),7.47–7.41(m,2H),7.29(d ,J=1.9Hz,1H),7.20(d,J=8.4Hz,1H),6.16(d,J=8.0Hz,1H),5.00–4.83(m,2H),4.32(dd,J=13.7,6.3Hz,1H),4.25–4.13(m,2H ),3.99(s,3H),3.11(d,J=11.4Hz,2H),2.98–2.62(m,8H),2.46(d,J=4.8Hz,2H),2.27(t,J=10.0Hz,2H),2.16–2.00(m,5H),1 .97–1.66(m,10H),1.53(dd,J=14.4,7.2Hz,4H),1.46–1.20(m,14H),1.03(d,J=14.4Hz,2H),0.87(t,J=7.4Hz,3H).HRMS(ESI) + :calcd for C 53 H 70 N 11 O7[M+H] + :972.5454,found:972.5453.

[0175] Example 7, 4-((R)-5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-N-(1-(11-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoisoindole-4-yl)undecyl)piperidin-4-yl)-3-methoxybenzamide

[0176]

[0177] Intermediate 1a (128 mg, 0.19 mmol) and 4-((11-bromondecyl)oxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (0.284 mmol) were dissolved in DMF (3 mL), and NaHCO3 (31 mg, 0.387 mmol) was added to the above solution. The above mixture was stirred at 65 ° C until the starting material was completely converted. The reaction mixture was quenched with 20 mL of water and extracted with 3×15 mL of EA. The combined organic layers were washed with 30 mL of saturated NaHCO3 and dried over MgSO4. After filtration and concentration under reduced pressure, the product was purified by flash column chromatography (PE / EA=3 / 1, v / v) to give a white solid (target product). (66 mg, 0.067 mmol, yield 35%) 1 H NMR (400MHz, CDCl3) δ8.47(dd,J=8.4,1.9Hz,1H),8.30(s,1H),7.80(d,J=1.6Hz,1H),7.66(dd,J=8.5,7.2Hz,1H),7.43(m,2H ),7.26(d,J=1.9Hz,1H),7.21(d,J=8.5Hz,1H),6.11(d,J=9.1Hz,1H),4.99–4.92(m,1H),4.88(dd,J=7.8,3.9Hz,1H),4.31(d d,J=13.7,6.2Hz,1H),4.19–4.14(m,2H),3.98(d,J=2.0Hz,3H),2.99(s,2H),2.92–2.73(m,8H),2.43–2.37(m,2H),2.24–2.0 1(m,8H),1.92–1.64(m,15H),1.55–1.49(m,4H),1.29–1.12(m,10H),1.03(d,J=13.3Hz,2H),0.86(t,J=7.4Hz,3H).HRMS(ESI) + :calcd for C 54 H 72 N 11 O7[M+H] + :986.5611,found:986.5615.

[0178] Example 8, 4-((R)-5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-N-(1-(4-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)acetamide)butyl)piperidin-4-yl)-3-methoxybenzamide

[0179]

[0180] Step 1: Synthesis of (R)-N-(1-(4-aminobutyl)piperidin-4-yl)-4-((5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-3-methoxybenzamide (8a)

[0181] Compound 1a (128 mg, 0.19 mmol) and tert-butyl (4-bromobutyl) carbamate (71 mg, 0.284 mmol) were dissolved in DMF (3 mL). NaHCO3 (31 mg, 0.387 mmol) was added to the above mixture and stirred at 70°C for 3 h. The reaction mixture was quenched with 20 ml of water and extracted with 3×15 ml of EA. The combined organic layers were washed with 15 ml of saturated NaHCO3 and dried over MgSO4. After filtration and concentration under reduced pressure, the product tert-butyl (R)-(4-((5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-3-methoxybenzamide piperidin-1-yl)butyl)carbamate was obtained, which was purified by flash column chromatography (PE / EA=4 / 1, v / v) to a white solid (109 mg, 0.15 mmol, 79% yield). It was then dissolved in DCM / TFA (V / V=2 / 1, 9 mL), and the mixture was stirred at room temperature for 3 hours. After the reaction was completed, the product was concentrated under reduced pressure to obtain a yellow solid (R)-N-(1-(4-aminobutyl)piperidin-4-yl)-4-((5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-3-methoxybenzamide (8a) (yield: 100%). ESI-MS [M+H]+ (M / z): 631.4.

[0182] Step 2: Synthesis of target product

[0183] Compound 8a (128 mg, 0.15 mmol) and 2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)oxyacetic acid (75 mg, 0.23 mmol) were dissolved in DCM (10 mL). EDCI (86 mg, 0.45 mmol), HOBT (81 mg, 0.60 mmol) and DIEA (58 mg, 0.45 mmol) were added to the above solution. The reaction mixture was stirred at room temperature for 12 hours. After concentration under reduced pressure, the product was purified by flash column chromatography (PE / EA=5 / 1, v / v) to a white solid (target product) (56 mg, 0.06 mmol, yield 39%). 1 H NMR (400MHz, CDCl3) δ8.46(d,J=8.4Hz,1H),8.29(s,1H),7.82–7.69(m,3H),7.55(d,J=7.3Hz,1H),7.45(d,J=1.9Hz,1H),7.3 1(dd,J=8.5,1.9Hz,1H),7.23(d,J=8.4Hz,1H),6.34(d,J=7.9Hz,1H),5.00–4.83(m,2H),4.71(d,J=14.2Hz,1H),4.66–4.58(m ,1H),4.31(dd,J=13.7,6.3Hz,1H),3.98(s,4H),3.52–3.43(m,1H),3.31(dd,J=13.3,5.7Hz,2H),3.12–2.97(m,3H),2.90–2. 69(m,8H),2.44(d,J=7.2Hz,2H),2.18(m,3H),2.05(s,2H),1.95–1.53(m,15H),1.02(m,2H),0.86(t,J=7.4Hz,3H).HRMS(ESI) + :calcd for C 49 H 61 N 12 O8[M+H] + :945.4730,found:945.4731.

[0184] Example 9, 4-((R)-5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-N-(1-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)acetyl)piperidin-4-yl)-3-methoxybenzamide

[0185]

[0186] Compound 1a (337 mg, 0.50 mmol) and 2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxyacetic acid (249 mg, 0.75 mmol) were dissolved in DCM (10 mL). EDCI (286 mg, 1.5 mmol), HOBT (270 mg, 2 mmol) and DIEA (193 mg, 1.5 mmol) were added to the above solution. The reaction mixture was stirred at room temperature for 12 hours. After concentration under reduced pressure, the product (target product) was purified by flash column chromatography (PE / EA=5 / 1, v / v). (174 mg, 0.20 mmol, yield 40%). 1 H NMR(400MHz, CDCl3)δ9.24(s,1H),8.45(dd,J=8.4,3.3Hz,1H),8.30(s,1H),7.80(d,J=10.9Hz,1H),7.66(m,1H),7.51–7 .41(m,2H),7.40–7.29(m,2H),6.69(dd,J=12.9,7.6Hz,1H),4.96(d,J=3.3Hz,1H),4.88(m,1H),4.49(d,J=12.5Hz,1H),4 .29(dd,J=14.3,6.1Hz,1H),4.24–4.01(m,2H),4.00–3.87(m,3H),3.21(q,J=11.9,11.4Hz,1H),2.97–2.68(m,8H),2.22– 2.08(m,4H),1.92(m,2H),1.74(m,6H),1.44–1.15(m,6H),1.01(p,J=11.4,10.7Hz,2H),0.85(t,J=7.4Hz,3H).HRMS(ESI) + :calcd for C 45 H 51 N 11 NaO8[M+Na] + :896.3814,found:896.3818.

[0187] Example 10, 4-((R)-5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-N-(1-(4-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)butyryl)piperidin-4-yl)-3-methoxybenzamide

[0188]

[0189] Step 1: Synthesis of (R)-N-(1-(4-bromobutyryl)piperidin-4-yl)-4-((5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-3-methoxybenzamide (10a)

[0190] Intermediate 1a (336 mg, 0.5 mmol) and 4-bromobutyric acid (0.75 mmol) were dissolved in DCM (10 mL). EDCI (286 mg, 1.5 mmol), HOBT (270 mg, 2 mmol) and DIEA (193 mg, 1.5 mmol) were added to the above solution. The reaction mixture was stirred at room temperature for 12 hours until the raw material was completely converted. After concentration under reduced pressure, it was purified by flash column chromatography (PE / EA=5 / 1, v / v) to give a white solid 10a (217 mg, 0.306 mmol, yield 61%). ESI-MS [M+H] + (M / z): 708.4.

[0191] Step 2: Synthesis of target product

[0192] Compound 10a (0.16mmol) and 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindoline-1,3-dione (65mg, 0.24mmol) were dissolved in N,N-dimethylformamide (3mL) solution. And NaHCO3 (26.5mg, 0.32mmol) and a catalytic amount of potassium iodide were added to the above solution. The mixture was stirred at 70°C for 6 hours. Then quenched with 15mL of water and extracted with 2×15mL EA. The combined organic phase was washed with 30mL of saturated NaHCO3 and dried with MgSO4. After filtration and concentration under reduced pressure, it was purified by flash column chromatography (PE / EA=4 / 1, v / v) to give a white solid product (target product). (53.3mg, 0.058mmol, yield 36%) 1H NMR (400MHz, CDCl3) δ8.47(dd,J=8.5,3.7Hz,1H),8.29(d,J=1.8Hz,1H),7.80(s,1H),7.71–7.64(m,1H),7.46( dd,J=6.3,2.9Hz,2H),7.30(s,1H),7.25(s,1H),6.48–6.25(m,1H),4.90(m,2H),4.59(d,J=13.2Hz,1H),4.30( dd,J=12.8,6.8Hz,4H),3.97(d,J=7.3Hz,3H),3.18(s,1H),2.77(s,8H),2.20(d,J=7.2Hz,2H),2.14–1.99(m,4 H),1.86–1.70(m,10H),1.48–1.38(m,2H),1.26(m,4H),1.00(d,J=9.3Hz,2H),0.86(t,J=7.4Hz,3H).HRMS(ESI) + :calcd for C 47 H 55 N 11 NaO8[M+Na] + :924.4127,found:924.4127.

[0193] Example 11, 4-((R)-5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-N-(1-(5-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)pentanoyl)piperidin-4-yl)-3-methoxybenzamide

[0194]

[0195] Step 1: Synthesis of (R)-N-(1-(5-bromovaleryl)piperidin-4-yl)-4-((5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-3-methoxybenzamide (11a)

[0196] Intermediate 1a (336 mg, 0.5 mmol) and 5-bromovaleric acid (0.75 mmol) were dissolved in DCM (10 mL). EDCI (286 mg, 1.5 mmol), HOBT (270 mg, 2 mmol) and DIEA (193 mg, 1.5 mmol) were added to the above solution. The reaction mixture was stirred at room temperature for 12 hours until the raw material was completely converted. After concentration under reduced pressure, it was purified by flash column chromatography (PE / EA=5 / 1, v / v) to give a white solid 11a (231 mg, 0.320 mmol, yield 64%). ESI-MS [M+H] + (M / z): 722.4.

[0197] Step 2: Synthesis of target product

[0198] Compound 11a (0.16mmol) and 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindoline-1,3-dione (65mg, 0.24mmol) were dissolved in N,N-dimethylformamide (3mL) solution. NaHCO3 (26.5mg, 0.32mmol) and a catalytic amount of potassium iodide were added to the above solution. The mixture was stirred at 70°C for 6 hours. It was then quenched with 15mL of water and extracted with 2×15mL of EA. The combined organic phase was washed with 30mL of saturated NaHCO3 and dried with MgSO4. After filtration and concentration under reduced pressure, it was purified by flash column chromatography (PE / EA=4 / 1, v / v) to give a white solid product (target product). (61mg, 0.067mmol, yield 42%). 1 H NMR(400MHz, CDCl3)δ8.46(dd,J=8.5,3.2Hz,1H),8.29(d,J=2.9Hz,1H),7.84–7.79(m,1H),7.68(m,1H),7.48–7 .39(m,2H),7.32(m,1H),7.21(d,J=8.5Hz,1H),6.48(dd,J=12.6,7.8Hz,1H),4.88(m,2H),4.57(d,J=13.5Hz,1H) ,4.33–4.14(m,4H),3.98(d,J=16.7Hz,3H),3.19(d,J=12.8Hz,1H),2.91–2.71(m,8H),2.60–2.46(m,2H),2.09–1 .92(m,10H),1.77–1.67(m,4H),1.42(m,2H),1.28–1.15(m,6H),1.09–0.96(m,2H),0.92–0.84(m,3H).HRMS(ESI) + :calcd for C 48 H57 N 11 NaO8[M+Na] + :938.4284,found:938.4288.

[0199] Example 12, 4-((R)-5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-N-(1-(6-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)hexyl)piperidin-4-yl)-3-methoxybenzamide

[0200]

[0201] Step 1: Synthesis of (R)-N-(1-(6-bromohexanoyl)piperidin-4-yl)-4-((5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-3-methoxybenzamide (12a)

[0202] Intermediate 1a (336 mg, 0.5 mmol) and 6-bromohexanoic acid (0.75 mmol) were dissolved in DCM (10 mL). EDCI (286 mg, 1.5 mmol), HOBT (270 mg, 2 mmol) and DIEA (193 mg, 1.5 mmol) were added to the above solution. The reaction mixture was stirred at room temperature for 12 hours until the raw material was completely converted. After concentration under reduced pressure, it was purified by flash column chromatography (PE / EA=5 / 1, v / v) to give a white solid 12a (198 mg, 0.269 mmol, yield 54%). ESI-MS [M+H] + (M / z): 736.4.

[0203] Step 2: Synthesis of target product

[0204] Compound 12a (0.16mmol) and 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindoline-1,3-dione (65mg, 0.24mmol) were dissolved in N,N-dimethylformamide (3mL) solution. NaHCO3 (26.5mg, 0.32mmol) and a catalytic amount of potassium iodide were added to the above solution. The mixture was stirred at 70°C for 6 hours. It was then quenched with 15mL of water and extracted with 2×15mL of EA. The combined organic phase was washed with 30mL of saturated NaHCO3 and dried with MgSO4. After filtration and concentration under reduced pressure, it was purified by flash column chromatography (PE / EA=4 / 1, v / v) to give a white solid product (target product). (64mg, 0.069mmol, yield 43%).1 H NMR(400MHz, CDCl3)δ9.29(s,1H),8.51–8.40(m,1H),8.30(s,1H),7.81(s,1H),7.74–7.64(m,1H),7.49–7.41(m,2H),7.33 (d,J=8.6Hz,1H),7.21(d,J=8.5Hz,1H),6.52(m,1H),4.89(m,2H),4.63(d,J=13.2Hz,1H),4.35–4.27(m,1H),4.18(q,J=5.9 Hz,2H),3.97(d,J=3.5Hz,3H),3.20(s,1H),2.78(d,J=15.3Hz,9H),2.40(q,J=6.2,5.1Hz,2H),2.18–2.01(m,4H),1.88(m,4 H),1.67–1.55(m,4H),1.45(q,J=11.7Hz,2H),1.33–1.12(m,10H),0.99(q,J=10.7Hz,2H),0.86(t,J=7.4Hz,3H).HRMS(ESI) + :calcd for C 49 H 59 N 11 NaO8[M+Na] + :952.4440,found:952.4446.

[0205] Example 13, 4-((R)-5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-N-(1-(7-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindole-4-yl)heptanoyl)piperidin-4-yl)-3-methoxybenzamide

[0206]

[0207] Step 1: Synthesis of (R)-N-(1-(7-bromoheptanoyl)piperidin-4-yl)-4-((5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-3-methoxybenzamide (13a)

[0208] Intermediate 1a (336 mg, 0.5 mmol) and 7-bromoheptanoic acid (0.75 mmol) were dissolved in DCM (10 mL). EDCI (286 mg, 1.5 mmol), HOBT (270 mg, 2 mmol) and DIEA (193 mg, 1.5 mmol) were added to the above solution. The reaction mixture was stirred at room temperature for 12 hours until the raw material was completely converted. After concentration under reduced pressure, it was purified by flash column chromatography (PE / EA=5 / 1, v / v) to give a white solid 13a. ESI-MS [M+H] + (M / z): 631.4.

[0209] Step 2: Synthesis of target product

[0210] Compound 13a (0.16mmol) and 2-(2,6-dioxopiperidin-3-yl)-4-hydroxyisoindoline-1,3-dione (65mg, 0.24mmol) were dissolved in N,N-dimethylformamide (3mL) solution. NaHCO3 (26.5mg, 0.32mmol) and a catalytic amount of potassium iodide were added to the above solution. The mixture was stirred at 70°C for 6 hours. It was then quenched with 15mL of water and extracted with 2×15mL of EA. The combined organic phase was washed with 30mL of saturated NaHCO3 and dried with MgSO4. After filtration and concentration under reduced pressure, it was purified by flash column chromatography (PE / EA=4 / 1, v / v) to give a white solid product (target product). (54mg, 0.057mmol, yield 36%). 1 H NMR (400MHz, CDCl3) δ8.47(d,J=8.5Hz,1H),8.30(s,1H),7.81(s,1H),7.66(dd,J=8.5,7.3Hz,1H),7.45(dd, J=11.3,5.9Hz,2H),7.31(dd,J=8.6,1.9Hz,1H),7.21(d,J=8.5Hz,1H),6.37(d,J=7.7Hz,1H),5.01–4.78(m,2 H),4.68–4.57(m,1H),4.30(dd,J=13.7,6.4Hz,1H),4.24–4.15(m,2H),3.97(d,J=1.8Hz,3H),3.27–3.16(m, 1H),2.94–2.65(m,8H),2.40–2.29(m,2H),2.11–1.20(m,27H),1.01(m,2H),0.86(t,J=7.4Hz,3H).HRMS(ESI) + :calcd for C 50 H 61 N 11 NaO8[M+Na]+ :966.4597,found:966.4601.

[0211] Example 14, 4-((R)-5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolo[4,3-f]pterin-7-yl)amino)-N-(1-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)glycyl)piperidin-4-yl)-3-methoxybenzamide

[0212]

[0213] Intermediate 1a (135 mg, 0.20 mmol) and (2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)glycine (0.30 mmol) were dissolved in DCM (10 mL) solution. EDCI (114 mg, 0.60 mmol), HOBT (108 mg, 0.80 mmol) and DIEA (77 mg, 0.60 mmol) were added to the above solution. The reaction mixture was stirred at room temperature for 12 hours. After concentration under reduced pressure, it was purified by flash column chromatography (PE / EA=4 / 1, v / v) to give a white solid (target product). (52 mg, 0.061 mmol, yield 30%). 1 H NMR (400MHz, CDCl3) δ9.31(d,J=16.7Hz,1H),8.66(d,J=9.3Hz,1H),8.48(d,J=8.4Hz,1H),8.29(s,1H),7.78(s,1H),7.66(t,J=7.8Hz, 2H), 7.44 (dd, J=13.6, 2.5Hz, 2H), 7.35 (dd, J=8.6, 1.9Hz, 1H), 6.55 (d, J=7.3Hz, 1H), 5.09 (dd, J=13.3, 5.1Hz, 1H), 4.87 (dd, J=7.9, 3. 9Hz,1H),4.39(d,J=5.3Hz,2H),3.95(s,3H),3.14(d,J=7.5Hz,1H),2.88–2.62(m,7H),2.40(t,J=7.3Hz,2H),2.31(t,J=7.2Hz,2H),2. 12(d,J=12.6Hz,2H),1.93(m,2H),1.63(d,J=47.3Hz,6H),1.36–1.21(m,9H),1.00(q,J=10.4Hz,2H),0.85(t,J=7.4Hz,3H).HRMS(ESI) + :calcd for C 45 H54 N 12 NaO6[M+Na] + :881.4181,found:881.4229.

[0214] Example 15, 4-((R)-5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolo[4,3-f]pterin-7-yl)amino)-N-(1-(5-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)piperidin-4-yl)-3-methoxybenzamide

[0215]

[0216] Intermediate 1a (135 mg, 0.20 mmol) and 5-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)pentanoic acid (0.30 mmol) were dissolved in DCM (10 mL) solution. EDCI (114 mg, 0.60 mmol), HOBT (108 mg, 0.80 mmol) and DIEA (77 mg, 0.60 mmol) were added to the above solution. The reaction mixture was stirred at room temperature for 12 hours. After concentration under reduced pressure, it was purified by flash column chromatography (PE / EA=4 / 1, v / v) to give a white solid (target product). (63 mg, 0.07 mmol, yield 35%). 1 H NMR (400MHz, CDCl3) δ8.52(dd,J=22.2,8.1Hz,2H),8.29(s,1H),7.78(s,1H),7.45(s,1H),7.39–7.28(m,2H),7.22(d,J=7.5Hz,1 H),6.76(d,J=8.0Hz,1H),6.31–6.15(m,1H),5.18(d,J=5.1Hz,1H),4.88(dd,J=8.0,3.9Hz,1H),4.61(d,J=13.5Hz,1H),4.37–4.2 5(m,2H),4.17(d,J=15.3Hz,2H),3.98(s,3H),3.86(d,J=13.6Hz,1H),3.72(q,J=7.0Hz,1H),3.27–3.13(m,3H),2.87–2.77(m,5H) ,2.43–2.29(m,3H),2.24–2.10(m,2H),1.71(d,J=12.4Hz,12H),1.39–1.13(m,8H),1.00(t,J=14.6Hz,2H),0.86(t,J=7.4Hz,3H).

[0217] Example 16, 4-((R)-5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-N-(1-(6-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)hexyl)piperidin-4-yl)-3-methoxybenzamide

[0218]

[0219] Intermediate 1a (135 mg, 0.20 mmol) and 6-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)hexanoic acid (0.30 mmol) were dissolved in DCM (10 mL) solution. EDCI (114 mg, 0.60 mmol), HOBT (108 mg, 0.80 mmol) and DIEA (77 mg, 0.60 mmol) were added to the above solution. The reaction mixture was stirred at room temperature for 12 hours. After concentration under reduced pressure, it was purified by flash column chromatography (PE / EA=4 / 1, v / v) to give a white solid (target product). (42 mg, 0.046 mmol, yield 23%). 1 H NMR (400MHz, CDCl3) δ8.47(d,J=8.4Hz,1H),8.29(s,1H),7.81(s,1H),7.47(s,1H),7.37(d,J=8.5Hz,1H),7.31(d,J=7.7H z,1H),7.17(d,J=7.5Hz,1H),6.75(d,J=8.1Hz,1H),6.63(m,1H),5.18–5.08(m,1H),4.87(dd,J=7.9,3.9Hz,1H),4.58(d, J=13.2Hz,1H),4.32–4.25(m,2H),4.22–4.14(m,2H),3.94(d,J=2.7Hz,3H),3.83(d,J=13.7Hz,1H),3.27–3.12(m,3H),2. 75(s,5H),2.32–1.99(m,9H),1.77–1.61(m,10H),1.46–1.15(m,10H),1.03–0.93(m,2H),0.85(t,J=7.4Hz,3H).HRMS(ESI) + :calcd for C 49 H 62 N 12 NaO6[M+Na] + :937.4807,found:937.4811.

[0220] Example 17, 4-((R)-5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-N-(1-(8-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindole-4-yl)amino)-8-oxooctyl)piperidin-4-yl)-3-methoxybenzamide

[0221]

[0222] Intermediate compound 1a (135 mg, 0.20 mmol) and 8-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)amino)-8-oxooctanoic acid (124 mg, 0.3 mmol) were dissolved in DCM (10 mL) solution. EDCI (114 mg, 0.60 mmol), HOBT (108 mg, 0.80 mmol) and DIEA (77 mg, 0.60 mmol) were added to the above solution. The reaction mixture was stirred at room temperature for 12 hours. After concentration under reduced pressure, the product (target product) (75 mg, 0.078 mmol, yield 39%) was purified by flash column chromatography (PE / EA=4 / 1, v / v). HRMS (ESI) + :calcd for C 51 H 64 N 12 NaO7[M+Na] + :979.4913,found:979.4911.

[0223] Example 18, 4-((R)-5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-N-(1-(2-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)ethoxy)ethyl)piperidin-4-yl)-3-methoxybenzamide

[0224]

[0225] Intermediate compound 1a (128 mg, 0.19 mmol) and 4-(2-(2-bromoethoxy)ethoxy)ethoxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (0.284 mmol) were dissolved in DMF (3 mL), and NaHCO3 (31 mg, 0.387 mmol) was added to the above solution. The above mixture was stirred at 65 ° C until the starting material was completely converted. The reaction mixture was quenched with 20 mL of water and extracted with 3×15 mL of EA. The combined organic layers were washed with 30 mL of saturated NaHCO3 and dried over MgSO4. After filtration and concentration under reduced pressure, the product was purified by flash column chromatography (PE / EA=3 / 1, v / v) to give a white solid (target product) (71.9 mg, 0.076 mmol, yield 40%). HRMS (ESI) + :calcd for C 49 H 62 N 11 O9[M+H] + :948.4726,found:948.4719.

[0226] Example 19, 4-((R)-5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-N-(1-(2-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)ethoxy)ethyl)piperidin-4-yl)-3-methoxybenzamide

[0227]

[0228] Intermediate compound 1a (128 mg, 0.19 mmol) and 3-(4-((2-(2-(2-bromoethoxy)ethoxy)ethyl)amino)-1-oxoisoquinolin-2-yl)piperidine-2,6-dione (0.284 mmol) were dissolved in DMF (3 mL), and NaHCO3 (31 mg, 0.387 mmol) was added to the above solution. The above mixture was stirred at 65°C until the starting material was completely converted. The reaction mixture was quenched with 20 mL of water and extracted with 3×15 mL of EA. The combined organic layers were washed with 30 mL of saturated NaHCO3 and dried over MgSO4. After filtration and concentration under reduced pressure, the product was purified by flash column chromatography (PE / EA=3 / 1, v / v) to give a white solid (target product) (63.7 mg, 0.068 mmol, yield 36%). HRMS (ESI) + :calcd for C 49 H 65 N 12 O7[M+H]+ :933.5094,found:933.5099.

[0229] Example 20, 4-((R)-5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-N-(1-(3-(2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)ethoxy)propanoyl)piperidin-4-yl)-3-methoxybenzamide

[0230]

[0231] Intermediate compound 1a (135 mg, 0.20 mmol) and 3-(2-(2-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoquinolin-4-yl)oxy)ethoxy)propanoic acid (130 mg, 0.3 mmol) were dissolved in DCM (10 mL) solution. EDCI (114 mg, 0.60 mmol), HOBT (108 mg, 0.80 mmol) and DIEA (77 mg, 0.60 mmol) were added to the above solution. The reaction mixture was stirred at room temperature for 12 hours. After concentration under reduced pressure, it was purified by flash column chromatography (PE / EA=4 / 1, v / v) to obtain a white solid product (target product) (83.8 mg, 0.086 mmol, yield 43%), HRMS (ESI) + :calcd for C 50 H 62 N 11 O 10 [M+H] + :976.4676,found:976.4686.

[0232] Example 21, 4-((R)-5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-N-(1-(3-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoxidolin-4-yl)amino)ethoxy)propanoyl)piperidin-4-yl)-3-methoxybenzamide

[0233]

[0234] Intermediate compound 1a (135 mg, 0.20 mmol) and 3-(2-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoquinolin-4-yl)amino)ethoxy)propionic acid (125 mg, 0.3 mmol) were dissolved in DCM (10 mL) solution. EDCI (114 mg, 0.60 mmol), HOBT (108 mg, 0.80 mmol) and DIEA (77 mg, 0.60 mmol) were added to the above solution. The reaction mixture was stirred at room temperature for 12 hours. After concentration under reduced pressure, the product (target product) (67 mg, 0.07 mmol, yield 35%) was purified by flash column chromatography (PE / EA=4 / 1, v / v). HRMS (ESI) + :calcd for C 50 H 65 N 12 O8[M+H] + :961.5034,found:961.5043.

[0235] Example 22, N-(4-((R)-5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-3-methoxyphenyl)-1-(7-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)heptyl)piperidine-4-carboxamide

[0236]

[0237] The intermediate compound (R)-N-(4-((5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-3-methoxyphenyl)piperidine-4-carboxamide (128 mg, 0.19 mmol) and 4-((7-bromoheptyl)oxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (127 mg, 0.284) were dissolved in DMF (3 mL), and NaHCO3 (31 mg, 0.387 mmol) was added to the above solution. The above mixture was stirred at 65°C until the starting material was completely converted. The reaction mixture was quenched with 20 mL of water and extracted with 3×15 mL of EA. The combined organic layers were washed with 30 mL of saturated NaHCO3 and dried over MgSO4. After filtration and concentration under reduced pressure, the product was purified by flash column chromatography (PE / EA=3 / 1, v / v) to give a white solid (target product) (72.3 mg, 0.078 mmol, yield 41%). HRMS (ESI) + :calcd for C 50 H64 N 11 O7[M+H] + :930.4985,found:930.4992.

[0238] Example 23, N-(4-((R)-5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-3-methoxyphenyl)-4-(7-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)heptyl)piperazine-1-carboxamide

[0239]

[0240] The intermediate compound (R)-N-(4-((5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-3-methoxyphenyl)piperazine-1-carboxamide (106 mg, 0.19 mmol) and 4-((7-bromoheptyl)oxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (127, 0.284) were dissolved in DMF (3 mL), and NaHCO3 (31 mg, 0.387 mmol) was added to the above solution. The above mixture was stirred at 65°C until the starting material was completely converted. The reaction mixture was quenched with 20 mL of water and extracted with 3×15 mL of EA. The combined organic layers were washed with 30 mL of saturated NaHCO3 and dried over MgSO4. After filtration and concentration under reduced pressure, the product was purified by flash column chromatography (PE / EA=3 / 1, v / v) to give a white solid (target product) (67 mg, 0.072 mmol, yield 38%). HRMS (ESI) + :calcd for C 49 H 63 N 12 O7[M+H] + :931.4937,found:931.4939.

[0241] Example 24, N-(4-((R)-5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-3-methoxyphenyl)-8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)octamide

[0242]

[0243] The intermediate compound (R)-N1-(5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolo[4,3-f]pterin-7-yl)-2-methoxybenzene-1,4-diamine (90 mg, 0.20 mmol) and 8-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)octanoic acid (124 mg, 0.3 mmol) were dissolved in a DCM (10 mL) solution. EDCI (114 mg, 0.60 mmol), HOBT (108 mg, 0.80 mmol) and DIEA (77 mg, 0.60 mmol) were added to the above solution. The reaction mixture was stirred at room temperature for 12 hours. After concentration under reduced pressure, the product was purified by flash column chromatography (PE / EA=4 / 1, v / v) to obtain a white solid product (target product) (74 mg, 0.088 mmol, yield 44%). HRMS (ESI) + :calcdfor C 45 H 55 N 10 O7[M+H] + :847.4250,found:847.4260.

[0244] Example 25, 4-((R)-5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-N-(7-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)heptyl)-3-methoxybenzamide

[0245]

[0246] The intermediate compound (R)-4-((5-(cyclohexylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-3-methoxybenzoic acid (95 mg, 0.20 mmol) and 4-((7-aminoheptyl)oxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (116 mg, 0.3 mmol) were dissolved in a DCM (10 mL) solution. EDCI (114 mg, 0.60 mmol), HOBT (108 mg, 0.80 mmol) and DIEA (77 mg, 0.60 mmol) were added to the above solution. The reaction mixture was stirred at room temperature for 12 hours. After concentration under reduced pressure, the product was purified by flash column chromatography (PE / EA=4 / 1, v / v) to obtain a white solid product (target product) (54 mg, 0.064 mmol, yield 32%). HRMS (ESI) +:calcdfor C 45 H 55 N 10 O7[M+H] + :847.4250,found:847.4256.

[0247] Example 26, 4-((R)-5-cyclopentyl-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-N-(1-(7-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)heptyl)piperidin-4-yl)-3-methoxybenzamide

[0248]

[0249] The intermediate compound (R)-4-((5-(cyclopentylmethyl)-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolo[4,3-f]pterin-7-yl)amino)-3-methoxy-N-(piperidin-4-yl)benzamide (104 mg, 0.19 mmol) and 4-((7-bromoheptyl)oxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (127, 0.284) were dissolved in DMF (3 mL) and NaHCO3 (31 mg, 0.387 mmol) was added to the above solution. The above mixture was stirred at 65°C until the starting material was completely converted. The reaction mixture was quenched with 20 mL of water and extracted with 3×15 mL of EA. The combined organic layers were washed with 30 mL of saturated NaHCO3 and dried over MgSO4. After filtration and concentration under reduced pressure, the product was purified by flash column chromatography (PE / EA=3 / 1, v / v) to give a white solid (target product) (73 mg, 0.079 mmol, yield 42%). HRMS (ESI) + :calcd for C 48 H 60 N 11 O7[M+H] + :902.4672,found:902.4677.

[0250] Example 27, N-(1-(7-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisooctanol-4-oxy)heptyl)piperidin-4-yl)-4-((R)-4-ethyl-5-isopropyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-3-methoxybenzamide

[0251]

[0252] The intermediate compound (R)-4-((4-ethyl-5-isopropyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-3-methoxy-N-(piperidin-4-yl)benzamide (96 mg, 0.19 mmol) and 4-((7-bromoheptyl)oxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (127, 0.284) were dissolved in DMF (3 mL), and NaHCO3 (31 mg, 0.387 mmol) was added to the above solution. The above mixture was stirred at 65°C until the starting material was completely converted. The reaction mixture was quenched with 20 mL of water and extracted with 3×15 mL of EA. The combined organic layers were washed with 30 mL of saturated NaHCO3 and dried over MgSO4. After filtration and concentration under reduced pressure, the product was purified by flash column chromatography (PE / EA=3 / 1, v / v) to give a white solid (target product) (59 mg, 0.068 mmol, yield 36%). HRMS (ESI) + :calcd for C 46 H 58 N 11 O7[M+H] + :876.4515,found:876.4519.

[0253] Example 28, 4-((R)-5-benzyl-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-N-(1-(7-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-4-yl)oxy)heptyl)piperidin-4-yl)-3-methoxybenzamide

[0254]

[0255] The intermediate compound (R)-4-((5-benzyl-4-ethyl-1-methyl-4,5-dihydro-[1,2,4]triazolo[4,3-f]pterin-7-yl)amino)-3-methoxy-N-(piperidin-4-yl)benzamide (106 mg, 0.19 mmol) and 4-((7-bromoheptyl)oxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (127, 0.284) were dissolved in DMF (3 mL), and NaHCO3 (31 mg, 0.387 mmol) was added to the above solution. The above mixture was stirred at 65°C until the starting material was completely converted. The reaction mixture was quenched with 20 mL of water and extracted with 3×15 mL of EA. The combined organic layers were washed with 30 mL of saturated NaHCO3 and dried over MgSO4. After filtration and concentration under reduced pressure, the product was purified by flash column chromatography (PE / EA=3 / 1, v / v) to give a white solid (target product) (68 mg, 0.074 mmol, yield 39%). HRMS (ESI) + :calcd for C 50 H 58 N 11 O7[M+H] + :924.4515found:924.4523.

[0256] Example 29, N-(1-(7-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoquinolin-4-yl)oxy)heptyl)piperidin-4-yl)-4-((R)-4-ethyl-1-methyl-5-(thien-2-ylmethyl)-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-3-methoxybenzamide

[0257]

[0258] The intermediate compound (R)-4-((4-ethyl-1-methyl-5-(thien-2-ylmethyl)-4,5-dihydro-[1,2,4]triazolo[4,3-f]pterin-7-yl)amino)-3-methoxy-N-(piperidin-4-yl)benzamide (106 mg, 0.19 mmol) and 4-((7-bromoheptyl)oxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (127, 0.284) were dissolved in DMF (3 mL), and NaHCO3 (31 mg, 0.387 mmol) was added to the above solution. The above mixture was stirred at 65°C until the starting material was completely converted. The reaction mixture was quenched with 20 mL of water and extracted with 3×15 mL of EA. The combined organic layers were washed with 30 mL of saturated NaHCO3 and dried over MgSO4. After filtration and concentration under reduced pressure, the product was purified by flash column chromatography (PE / EA=3 / 1, v / v) to give a white solid (target product) (58 mg, 0.062 mmol, yield 33%), HRMS (ESI): + :calcd for C 48 H 56 N 11 O7S[M+H] + :930.4079,found:930.4080.

[0259] Example 30, N-(1-(7-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoquinolin-4-yl)oxy)heptyl)piperidin-4-yl)-4-((R)-4-ethyl-1-methyl-5-(pyridin-2-ylmethyl)-4,5-dihydro-[1,2,4]triazolyl[4,3-f]pterin-7-yl)amino)-3-methoxybenzamide

[0260]

[0261] The intermediate compound (R)-4-((4-ethyl-1-methyl-5-(pyridin-2-ylmethyl)-4,5-dihydro-[1,2,4]triazolo[4,3-f]pterin-7-yl)amino)-3-methoxy-N-(piperidin-4-yl)benzamide (106 mg, 0.19 mmol) and 4-((7-bromoheptyl)oxy)-2-(2,6-dioxopiperidin-3-yl)isoindoline-1,3-dione (127, 0.284) were dissolved in DMF (3 mL) and NaHCO3 (31 mg, 0.387 mmol) was added to the above solution. The above mixture was stirred at 65°C until the starting material was completely converted. The reaction mixture was quenched with 20 mL of water and extracted with 3×15 mL of EA. The combined organic layers were washed with 30 mL of saturated NaHCO3 and dried over MgSO4. After filtration and concentration under reduced pressure, the product was purified by flash column chromatography (PE / EA=3 / 1, v / v) to give a white solid (target product) (70 mg, 0.076 mmol, yield 40%). HRMS (ESI) + :calcd for C 49 H 57 N 12 O7[M+H] + :925.4468,found:925.4480.

[0262] The beneficial effects of the present invention are demonstrated below through specific test examples.

[0263] Experimental Example 1, Examples 1-30, Inhibitory Activity on Tumor Cell Proliferation

[0264] Experimental method: Tumor cells in the logarithmic growth phase were collected and resuspended in the corresponding cell culture medium to 1-9×10 5 / mL, add the cell suspension to the 96-well plate at a volume of 100μL per well, and fill the side wells with 200μL sterile saline. Incubate the adherent cells in a 37°C incubator for 24 to 48 hours to allow the cells to adhere, and the suspended cells can be added with drugs immediately. Prepare the specified compound in the deep-well plate dosing tank with the corresponding culture medium to prepare a drug-containing culture medium with a specified concentration of the compound, and add it to the 96-well plate at a volume of 100μL per well. Continue incubating in a 37°C incubator for 4 days, add 20μL of 5% MTT solution to each well under light-proof conditions, and continue incubating in the incubator for 2 to 4 hours. Afterwards, for suspended cells, add 50μL of 20% SDS solution to each well and place it in the incubator overnight; for adherent cells, carefully discard the liquid in the well, and then add 150μL of dimethyl sulfoxide to each well; use an enzyme reader to measure the absorbance value at a wavelength of 570nM, use GraphPad 8.0 software to draw a cell growth curve, and calculate the IC 50 value.

[0265] The inhibitory activity of all the compounds in the examples on MV4-11 cell line (IC 50 ) are as follows, where " / " means not tested, and the test results are shown in Table 1. (where "+" means IC 50 >10μM; “++” means 10μM>IC 50 >1μM; “+++” indicates IC 50 <1 μM).

[0266] Table 1. Proliferation inhibition activity of compounds of Examples 1-30 on MV4-11 cells

[0267]

[0268]

[0269] The proliferation inhibition activity (IC 50 ) are as follows, where “ / ” means not tested, and the test results are shown in Table 2.

[0270] Table 2. Proliferation inhibition activity of the compounds of Example 3 and Example 9 on various cells

[0271]

[0272] Experimental Example 2: Effects of Examples 3 and 9 on the expression levels of some proteins

[0273] Test method:

[0274] ① Cell culture and drug treatment: Tumor cells were inoculated at an appropriate density in a 25 mm 2 The cells were cultured in a culture dish in a 37°C incubator for 24 to 48 hours to allow the cells to adhere to the wall, after which the upper culture medium was discarded and the cells were treated with fresh culture medium containing the specified concentration of the compound for the corresponding time.

[0275] ②Total protein extraction: discard the old culture medium, digest the cells with trypsin, collect the cell precipitate by centrifugation, wash the cells twice with PBS buffer, add an appropriate amount of RIPA lysis buffer (containing cocktail 1:1000) according to the amount of cell precipitate, place on ice, lyse the cells for 40 minutes, then ultrasonically disrupt the cells on ice (3 seconds / time, 1 second interval, 3 times in total), then lyse on ice for 30 minutes, 12000rpm, 4℃, centrifuge for 15 minutes, and transfer the supernatant to a new EP tube.

[0276] ③BCA quantification: Use the BCA protein quantification kit to quantify the total protein, and dilute the sample with RIPA to make the total protein concentration of each sample the same, then add 5× protein loading buffer, place in a metal bath at 100°C for ten minutes to denature the protein sample, and store the prepared sample at -20°C.

[0277] ④SDS-PAGE gel electrophoresis and immunoblotting analysis: First, prepare SDS-PAGE gel and electrophoresis buffer of different concentrations (7.5% to 12.5%) according to the molecular weight of the target protein, clamp the gel plate in the electrophoresis tank, check for leakage with ultrapure water, pour out the ultrapure water after confirmation, and add electrophoresis buffer. Add protein pre-stained marker and protein sample in the order of loading, adjust the voltage to 80V, and start electrophoresis. Wait for about 20 minutes, after the sample runs through the concentrated gel and enters the separation gel, adjust the voltage to 120V, and continue electrophoresis until the sample runs to the bottom of the gel plate. After the electrophoresis is finished, pour the prepared transfer buffer into the disk, take out the gel plate, gently scrape off the upper concentrated gel with a scraper, and make a transfer "sandwich" of NC membrane, filter paper and gel in the order of (red side) sponge → filter paper → NC membrane → gel → filter paper → sponge (black side), and quickly insert it into the transfer clip (carefully remove the bubbles between the layers with a roller), and transfer the membrane at 100V for 1 to 2.5 hours. After the transfer is completed, take out the NC membrane and gently rinse it in TBS / T buffer to remove methanol. Then put the NC membrane into the milk blocking solution and block it at room temperature on a shaker for 1 hour. After the blocking is completed, gently rinse it in TBS / T buffer, cut the membrane according to the molecular weight, and incubate it in the primary antibody prepared in advance with the primary antibody diluent at 4° overnight. Take out the band and wash the band with TBS / T buffer for 10 minutes / time, for a total of three washes. Then, incubate with the corresponding HRP secondary antibody at room temperature for one hour, and then wash the strips with TBS / T buffer for 10 minutes each time, for a total of three times. Finally, add developer to the strips and expose them on an exposure machine.

[0278] The effects of Example 3 and Example 9 on BRD2 / 3 / 4, PLK1, and c-Myc protein levels are shown in Figure 1 Example 9 Effects of different durations on BRD4 and c-Myc protein levels in various prostate cancer cells Figure 2 The effects of different concentrations of Example 9 on BRD4 and c-Myc protein levels in various prostate cancer cells after 24 hours are shown in Figure 3As can be seen from the figure, Examples 3 and 9 have a selective down-regulation effect on BRD4 and its downstream protein c-Myc, but have no significant effect on BRD2 / 3 and PLK1, indicating that the embodiments of the present invention have a high selectivity for down-regulation of BRD4 protein. In view of the fact that BRD2 / 3 / 4 and PLK1 are not only related to the occurrence of tumors, but also participate in the regulation of various physiological activities of normal cells, small molecules that have a selective degradation effect on BRD4 should have better safety when used in disease treatment than protein degradation molecules that can simultaneously degrade BRD4 and PLK1.

[0279] In summary, the compounds of the present invention, or their stereoisomers, or their pharmaceutically acceptable salts, or their solvates can induce the degradation of BRD4 protein, thereby exerting a therapeutic effect on BRD4-dependent tumors or other diseases. Such compounds are expected to be used as one of the ingredients of pharmaceutical preparations for the treatment of BRD4-dependent tumors or other diseases.

Claims

1. A compound or a salt thereof, characterized in that The compound is shown in formula (III): in, R1 is selected from substituted methyl; The substituent of the methyl group in R1 is selected from phenyl; A is selected from X is selected from -CH2-, -C(O)-; L is selected from -Ak2-Z-Ak1-Y-; Y is selected from -O-, -NR 16 - or None; Z is selected from -O-, -NR 17 or none; R 16 With R 17 are independently selected from hydrogen, -C 1-6 alkyl; Ak1 is selected from -C(O)(CH2) n3 -、-(CH2) n4 C(O)-、-(CH2) n5 -、-(CH2CH2O) n10 CH2CH2- or none; Ak2 is selected from -C(O)(CH2) n6 -、-(CH2) n7 -、-(CH2CH2O) n8 -、-(CH2CH2O) n9 CH2CH2- or none; n3, n4, n5, n6, and n7 are each independently selected from an integer between 0 and 15; n8, n9, and n10 are each independently selected from integers between 0 and 5.

2. The compound or salt thereof according to claim 1, characterized in that: L is selected from -Ak2-Z-Ak1-Y-; Y is selected from -O-, -NH- or none; Z is selected from -O-, -NH- or none; Ak1 is selected from -C(O)(CH2) n3 -、-(CH2) n4 C(O)-、-(CH2) n5 -、-(CH2CH2O) n10 CH2CH2- or none; Ak2 is selected from -C(O)(CH2) n6 -、-(CH2) n7 -、-(CH2CH2O) n8 -、-(CH2CH2O) n9 CH2CH2- or none; n3, n4, n5, n6, and n7 are each independently selected from an integer between 0 and 15; n8, n9, and n10 are each independently selected from integers between 0 and 5.

3. The compound or salt thereof according to claim 2, characterized in that: L is selected from n is selected from an integer between 0-10.

4. The compound or salt thereof according to any one of claims 1 to 3, characterized in that: The compound is shown in formula (IV): in, R1, L and X are selected from any one of claims 1 to 3.

5. The compound or salt thereof according to any one of claims 1 to 3, characterized in that: The compound is shown in formula (V): in, L and X are selected from any one of claims 1 to 3.

6. The compound or salt thereof according to any one of claims 1 to 3, characterized in that: The compound is one of the following compounds:

7. Use of the compound or salt thereof according to any one of claims 1 to 6 in the preparation of BRD4 protein degrading agents.

8. The use according to claim 7, wherein the drug is a drug that has a down-regulating effect on BRD4 and / or its downstream protein c-Myc.

9. The use according to claim 7, characterized in that: The drug is a drug for treating BRD4-dependent tumors, diabetes, atherosclerosis, and coronary artery disease.

10. The use according to claim 9, characterized in that: The tumor is leukemia, lymphoma, liver cancer, breast cancer, colon cancer, rectal cancer, colorectal cancer, lung cancer, and prostate cancer.

11. A pharmaceutical composition, characterized in that: The invention relates to a preparation prepared by taking the compound or salt thereof as the active ingredient and adding pharmaceutically acceptable auxiliary materials.

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