Heterocyclic compound as well as preparation method and application thereof

Abstract

The invention provides a heterocyclic compound as well as a preparation method and application thereof, and belongs to the technical field of biological medicines. The structure of the heterocyclic compound is shown as a formula I. The compound disclosed by the invention has good inhibitory activity on a PI3K target spot and an HDAC target spot at the same time, and can be used as a double-target spot inhibitor with an excellent effect. The compound provided by the invention can be used for preparing medicines for preventing and / or treating diseases related to PI3K and / or HDAC, and has a good application prospect. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and specifically relates to a heterocyclic compound and a preparation method and use thereof. Background Art

[0002] Malignant tumors are one of the most serious public health problems in the world. Although the emergence of new therapeutic drugs and treatment methods in recent years has significantly improved the quality of life of cancer patients, overall, the current treatment status of malignant tumors is far from satisfactory, and more efforts are urgently needed to find new anti-tumor drugs.

[0003] Chemotherapy has always played an important role in the treatment of tumors due to its wide adaptability and thorough elimination of tumor cells. 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. Tumor targeted drugs represented by kinase inhibitors have revolutionized the treatment of malignant tumors in the past 20 years due to their advantages such as rapid efficacy and mild toxic side effects. However, due to the rapid generation of drug-resistant mutations, these drugs can only control tumor growth in a relatively short period of time. 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 defects of traditional kinase inhibitors and bring a new revolution to the treatment of malignant tumors. However, these inhibitors also have shortcomings such as insufficient indications and slow efficacy. Due to the complementary advantages of kinase inhibitors and epigenetic targets, small molecule compounds that can act on kinase targets and epigenetic targets at the same time are expected to become a new type of highly effective anti-tumor drugs.

[0004] Abnormal activation or amplification of PI3K is closely related to the occurrence and development of hematological malignancies. Inhibitors targeting this protein can be used to treat hematological tumors. Currently, a large number of PI3K inhibitors have been reported, but the clinical indications of existing PI3K inhibitors are relatively limited, and their therapeutic effects are far from satisfactory due to disease recurrence and drug resistance.

[0005] Histone deacetylase (HDAC) is a type of protease that plays an important role in the structural modification of chromosomes and the regulation of gene expression. This protein catalyzes the deacetylation of histones to make DNA and histone octamers more tightly bound, thereby inhibiting the transcription of genes, especially tumor suppressor genes. The overexpression or abnormal activation of this protein is closely related to the occurrence and development of various tumors. Therefore, several small molecule inhibitors targeting this type of protein, such as Vorinostat, Romidepsin, Belinostat, etc., have been approved for the treatment of cutaneous T-cell lymphoma (CTCL), peripheral T-cell lymphoma (PTCL) and multiple myeloma (MM). In addition, there are currently a large number of HDAC inhibitors in different clinical stages for the treatment of various hematological tumors and solid tumors. However, the efficacy of current HDAC inhibitors in tumor patients is still unsatisfactory, and most patients can only achieve a few months of disease remission and prolonged survival through the treatment of this type of drug.

[0006] Both PI3K and HDAC single-target inhibitor drugs have certain limitations due to insufficient efficacy and rapid generation of drug-resistant mutations. Studies have found that simultaneous inhibition of PI3K and HDAC can synergistically inhibit tumor growth and provide a better therapeutic range than single inhibitors by improving efficacy and limiting drug resistance. Currently, multiple PI3K / HDAC dual-target inhibitors have been reported (DOI: 10.1021 / acs.jmedchem.9b00390), such as CUDC-907, which inhibits cancer cell proliferation through synergistic effects and has better efficacy than single-target inhibitors, but it still cannot meet clinical needs. There is an urgent need to develop more inhibitors that can simultaneously inhibit PI3K and HDAC. In view of this, this invention is specially proposed. Summary of the invention

[0007] The purpose of the present invention is to provide a heterocyclic compound and a preparation method and use thereof. The heterocyclic compound of the present invention can be used as a novel dual-target inhibitor, capable of simultaneously inhibiting kinase targets and epigenetic targets, and can be used to prepare highly effective anti-tumor drugs.

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

[0009]

[0010]

[0011] in,

[0012] R1 is LR a R b CONHR c ,

[0013] L is selected from none, C1-C6 alkylene, carbonyl;

[0014] R a Select from None, C1~C 10 Alkylene, 6- to 10-membered aryl which is unsubstituted or substituted by one or more R', 5- to 10-membered heteroaryl which is unsubstituted or substituted by one or more R';

[0015] R b Select from None, C2~C 10 alkenylene;

[0016] R c is selected from hydroxyl, 6-10 membered aryl which is unsubstituted or substituted by one or more R';

[0017] And L, R a , R b Not at the same time is nothing;

[0018] R2, R3, R4, R5, R6, R7, R8, and R9 are independently selected from hydrogen, or two substituents on the same carbon atom form =O, and at most one of R2, R3, R4, R5, R6, R7, R8, and R9 forms =O;

[0019] R 10 Selected from C1-C6 alkyl groups which are unsubstituted or substituted by one or more R';

[0020] R 11 is selected from a cyclic ring which is unsubstituted or substituted with one or more R';

[0021] R' is independently selected from hydroxy, halogen, cyano, amino, C1-C6 alkyl, C2-C6 alkenyl, C1-C6 alkoxy, and C2-C6 alkynyl.

[0022] Further, the compound is as shown in Formula II:

[0023]

[0024] in,

[0025] R1' is selected from C1-C6 alkyl;

[0026] R1 is LR a R b CONHR c ,

[0027] L is selected from none, C1-C6 alkylene, carbonyl;

[0028] R a Select from None, C1~C 10Alkylene, 6- to 10-membered aryl which is unsubstituted or substituted by one or more R', 5- to 10-membered heteroaryl which is unsubstituted or substituted by one or more R';

[0029] R b Select from None, C2~C 10 alkenylene;

[0030] R c is selected from hydroxyl, 6-10 membered aryl which is unsubstituted or substituted by one or more R';

[0031] And L, R a , R b Not at the same time is nothing;

[0032] R2, R3, R4, R5, R6, R7, R8, and R9 are independently selected from hydrogen, or two substituents on the same carbon atom form =O, and at most one of R2, R3, R4, R5, R6, R7, R8, and R9 forms =O;

[0033] R 10 Selected from C1-C6 alkyl groups which are unsubstituted or substituted by one or more R';

[0034] R' is independently selected from hydroxy, halogen, cyano, amino, C1-C6 alkyl, C2-C6 alkenyl, C1-C6 alkoxy, and C2-C6 alkynyl.

[0035] Further, R1' is selected from C1-C6 alkyl;

[0036] Selected from

[0037] R1 is LR a R b CONHR c ,

[0038] L is selected from none, C1-C6 alkylene, carbonyl;

[0039] R a Select from None, C1~C 10 Alkylene, 6- to 10-membered aryl which is unsubstituted or substituted by one or more R', 5- to 10-membered heteroaryl which is unsubstituted or substituted by one or more R';

[0040] R b Select from None, C2~C 10 alkenylene;

[0041] R c is selected from hydroxyl, 6-10 membered aryl which is unsubstituted or substituted by one or more R';

[0042] And L, R a , R b Not at the same time is nothing;

[0043] R 10 Selected from C1-C6 alkyl groups which are unsubstituted or substituted by one or more R';

[0044] R' is independently selected from amino, C1-C6 alkyl, and C2-C6 alkenyl.

[0045] Furthermore, the compound is as shown in Formula III, Formula IV, and Formula V:

[0046]

[0047] in,

[0048] R1' is selected from C1-C3 alkyl;

[0049] R 10 Selected from C1-C3 alkyl;

[0050] R1 is LR a R b CONHR c ,

[0051] L is selected from none, C1-C3 alkylene, carbonyl;

[0052] R a Selected from none, C1-C8 alkylene, phenyl;

[0053] R b Selected from none, C2-C3 alkenylene;

[0054] And L, R a , R b Not at the same time is nothing;

[0055] R c is selected from hydroxyl, phenyl which is unsubstituted or substituted by one or more R';

[0056] R' is each independently selected from amino;

[0057] Preferably, R1 is Furthermore, the compound is one of the following compounds:

[0058]

[0059]

[0060] The present invention also provides use of the above-mentioned compound, its salt, its stereoisomer or its solvate in the preparation of PI3K inhibitors and / or HDAC inhibitors.

[0061] Furthermore, the PI3K inhibitor is a PI3Kδ inhibitor, and the HDAC inhibitor is an HDAC6 inhibitor.

[0062] The present invention also provides the use of the above-mentioned compound, its salt, its stereoisomer or its solvate in the preparation of a drug for preventing and / or treating diseases related to PI3K and / or HDAC.

[0063] Furthermore, the disease associated with PI3K and / or HDAC is cancer, and the cancer is preferably lymphoma.

[0064] The present invention also provides a pharmaceutical composition, which is a preparation prepared by taking the above-mentioned compound, its salt, its stereoisomer or its solvate as an active ingredient and adding pharmaceutically acceptable excipients or auxiliary ingredients.

[0065] 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.

[0066] 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.

[0067] "Substitution" refers to the replacement of a hydrogen atom in a molecule by another different atom or molecule.

[0068] The minimum and maximum carbon atom content in a hydrocarbon group is indicated by a prefix, for example, the prefix C a ~C b Alkyl refers to any alkyl group containing from "a" to "b" carbon atoms. Thus, for example, "C1-C6 alkyl" refers to an alkyl group containing 1, 2, 3, 4, 5, or 6 carbon atoms. For example, C 1-6 Alkyl refers to straight or branched chain alkyl groups containing 1, 2, 3, 4, 5 or 6 carbon atoms, and so on.

[0069] "Alkyl" refers to a saturated hydrocarbon chain having a specified number of carbon atoms. For example, C1-C6 alkyl refers to an alkyl group having 1 to 6 carbon atoms, i.e., 1, 2, 3, 4, 5 or 6 carbon atoms. Alkyl groups can be straight or branched. Representative branched alkyl groups have one, two or three branches. Alkyl groups include methyl, ethyl, propyl (n-propyl and isopropyl), butyl (n-butyl, isobutyl and tert-butyl), pentyl (n-pentyl, isopentyl and neopentyl) and hexyl, etc.

[0070] "Alkenyl" refers to an aliphatic hydrocarbon group having at least one carbon-carbon double bond. The alkenyl group may be straight chain or branched.

[0071] "Alkynyl" refers to an aliphatic hydrocarbon group having at least one carbon-carbon triple bond. The alkynyl group may be straight chain or branched.

[0072] "Aryl" refers to an all-carbon monocyclic group with a conjugated π electron system, such as phenyl. The aryl group does not contain heteroatoms, such as nitrogen, oxygen, or sulfur, and the point of attachment to the parent must be on a carbon atom on the ring with a conjugated π electron system.

[0073] "Heteroaryl" or "heteroaromatic ring" refers to a heteroaromatic group containing one to multiple heteroatoms. The heteroatoms referred to herein include, but are not limited to, oxygen, sulfur, and nitrogen. For example, furanyl, thienyl, pyridyl, pyrazolyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, imidazolyl, tetrazolyl, etc.

[0074] 5-10 membered heteroaryl refers to heteroaromatic groups containing 5, 6, 7, 8, 9 or 10 ring atoms, and so on.

[0075] "Joint ring" refers to a polycyclic ring in which two rings share two adjacent ring atoms.

[0076] "Halogen" refers to fluorine, chlorine, bromine or iodine.

[0077] The present invention provides a heterocyclic compound, which has good inhibitory activity against both PI3K and HDAC targets, and can be used as an excellent dual-target inhibitor, thereby effectively inhibiting the proliferation of tumor cells. This dual-target inhibitor can complement the advantages of the two targets, synergistically exert anti-tumor effects, have a wider range of indications and better efficacy, and reduce the occurrence of drug resistance.

[0078] 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.

[0079] The above contents of the present invention are further described in detail below 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 realized based on the above contents of the present invention belong to the scope of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0080] Figure 1 This is the H NMR spectrum of compound Z1.

[0081] Figure 2 This is the carbon NMR spectrum of compound Z1.

[0082] Figure 3 This is the H NMR spectrum of compound Z2.

[0083] Figure 4 This is the carbon NMR spectrum of compound Z2.

[0084] Figure 5 This is the H NMR spectrum of compound Z3.

[0085] Figure 6 This is the carbon NMR spectrum of compound Z3.

[0086] Figure 7 This is the H NMR spectrum of compound Z4.

[0087] Figure 8 This is the carbon NMR spectrum of compound Z4.

[0088] Fig. 9 This is the H NMR spectrum of compound Z5.

[0089] Fig.10 This is the carbon NMR spectrum of compound Z5.

[0090] Fig.11 This is the H NMR spectrum of compound Z6.

[0091] Fig.12 This is the carbon NMR spectrum of compound Z6.

[0092] Fig.13 This is the H NMR spectrum of compound Z8.

[0093] Fig.14 This is the carbon NMR spectrum of compound Z8.

[0094] Fig.15 This is the H NMR spectrum of compound Z9.

[0095] Fig.16 This is the carbon NMR spectrum of compound Z9.

[0096] Fig.17 This is the H NMR spectrum of compound Z10.

[0097] Fig.18 This is the carbon NMR spectrum of compound Z10. DETAILED DESCRIPTION

[0098] 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.

[0099] The "room temperature" or "normal temperature" referred to in the present invention is 25±10°C.

[0100] Example 1

[0101] (E)-3-(4-(4-(2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-oxopiperazin-1-yl)methylphenyl)-N-hydroxyacrylamide (Compound Z1)

[0102] Step 1: Preparation of tert-butyl 4-((2-chloro-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-oxopiperazine-1-carboxylate (Intermediate 1)

[0103]

[0104] Boc piperazinone (4.15 g, 16.6 mmol) was added to THF (10 ml), and the mixture was dissolved by stirring at ℃. Then t-BuOK (3.5 g, 31.13 mmol) was added, and the mixture was stirred at 0℃ for 10 min. Then, the raw material 4-(2-chloro-8-(chloromethyl)-9-methyl-9H-purin-6-yl)morpholine (5 g, 16.6 mmol) was added, and the mixture was stirred at 0℃ for 20 min. Then, the mixture was stirred at room temperature for 1 h. The reaction progress was monitored by TLC. After the raw material was completely converted, water was added, the product was precipitated, and the product was filtered. The filter cake was taken and dried to obtain 4.9 g of the product with a yield of 63.45%.

[0105] Step 2: Preparation of tert-butyl 4-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-oxopiperazine-1-carboxylate (Intermediate 2)

[0106]

[0107] Intermediate 1 (4.3 g, 9.24 mmol), raw material 2-ethyl-1H-benzo[d]imidazole (1.35 g, 9.24 mmol), Pd2(dba)3 (0.96 g, 0.92 mmol), Xpos (0.88 g, 1.85 mmol), CsCO3 (6 g, 18.48 mmol) were added to 1,4-dioxane (50 ml), and the mixture was ventilated with nitrogen for 3 times under stirring. The reaction was stirred at 110°C for 3 h under nitrogen protection. The reaction progress was monitored by TLC. The raw material was completely converted, and the mixture was extracted with DCM, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 4.5 g of the product with a yield of 85%.

[0108] Step 3: Preparation of 1-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)piperazin-2-one hydrochloride (Intermediate 3)

[0109]

[0110] To the intermediate 2 (4.5 g, 7.82 mmol), add dioxane hydrochloride solution (50 ml), and react at room temperature with stirring for 2 h. The reaction progress was monitored by TLC until Boc was completely removed. DCM was added to the reaction solution, and ultrasonication was performed. The liquid was poured out and washed with DCM twice. MeOH was then added to dissolve the solid part, and the product was concentrated under reduced pressure to obtain 3.12 g of product with a yield of 78%.

[0111] Step 4: Preparation of 1-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)piperazin-2-one (Intermediate 4)

[0112]

[0113] Add saturated potassium carbonate solution (30 ml) to intermediate 3 (3.12 g, 6.1 mmol), shake and react, then extract with DCM, wash with saturated brine, dry over anhydrous sodium sulfate, and concentrate under reduced pressure. Dissolve in DCM, add silica gel to mix, load onto column, and flush out 2.5 g of product with DCM:MeOH from 100:1 to 10:1, with a yield of 85%.

[0114] Step 5: Preparation of (E)-methyl 3-(4-(4-(2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-oxopiperazin-1-yl)methylphenyl)acrylate (Intermediate 5)

[0115]

[0116] The intermediate 4 (500 mg, 1.05 mmol) was dissolved in acetonitrile solution (10 ml), and methyl 4-bromomethylcinnamate (268 mg, 1.05 mmol) and potassium carbonate (435 mg, 3.15 mmol) were added. The mixture was heated and stirred at 85°C for 2 h. The reaction progress was monitored by TLC. After the reaction was complete, the reaction solution was dried by spin drying, extracted with DCM, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 654 mg of the product with a yield of 86%.

[0117] Step 6: Preparation of (E)-3-(4-(4-(2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-oxopiperazin-1-yl)methylphenyl)acrylic acid (Intermediate 6)

[0118]

[0119] Dissolve the intermediate 5 (650 mg, 1 mmol) in a mixed solution of THF (8 ml), MeOH (8 ml), and H2O (2 ml), then add LiOH·H2O (210 mg, 5 mmol), stir at 70°C, and react for 1 h. Monitor the reaction progress by TLC. After the reaction is complete, spin dry the solvent in the reaction solution, remove THF and MeOH, add a small amount of water, and then add HCl to adjust the acidity under stirring until a precipitate is precipitated. The pH test paper shows weak acidity. Wash the solid part with ethyl acetate (EA) 3 times, dissolve the solid part with DCM and MeOH, and concentrate under reduced pressure to obtain 495 mg of the product with a yield of 78%.

[0120] Step 7: Preparation of (E)-3-(4-((4-(2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-oxopiperazin-1-yl)methyl)phenyl)-N-(((tetrahydro-2H-pyran-2-yl)oxy)acrylamide (Intermediate 7)

[0121]

[0122] The intermediate 6 (200 mg, 0.31 mmol) was dissolved in DMF (5 ml), and HATU (178 g, 0.47 mmol) and DIEA (60 mg, 0.47 mmol) were added, followed by the raw material O-(tetrahydro-2H-pyran-2-yl)hydroxylamine (36 mg, 0.31 mmol). The mixture was stirred at room temperature for 30 min. After the reaction was complete, a small amount of water was added, and the mixture was extracted twice with EA, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 315 mg of a crude product. After plate purification, 180 mg of the product was obtained with a yield of 79%.

[0123] Step 8: Preparation of compound Z1

[0124]

[0125] To the intermediate 7 (62 mg, 0.084 mmol), 6 mol / L HCl aqueous solution (4 ml) was added, the mixture was stirred at room temperature, and the reaction was carried out for 30 min. The reaction progress was monitored by TLC. After the reaction was complete, sodium bicarbonate solution was used for alkali adjustment. When a white solid was precipitated and pH paper detected that it was alkaline, the reaction solution was filtered, and the filter cake was dissolved in DCM and MeOH, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 38.7 mg of the product with a yield of 70%.

[0126] Characterization data: 1H NMR (600MHz, DMSO-d6) δ8.27–8.18(m,1H),7.94–7.84(m,1H),7.78–7.55(m,6H),7.46(d,J=15.8Hz,1H),6.62(d,J=15.8Hz,1H),5.11(s,2 H), 4.91–4.62 (m, 2H), 4.48 (s, 2H), 4.04 (s, 2H), 3.78 (d, J = 9.2Hz, 9H), 3.67 (d, J = 2.1Hz, 2H), 3.52 (d, J = 7.5Hz, 4H), 1.48 (t, J = 7.4Hz, 3H).

[0127] HRMS (ESI+) m / z theoretical value: C 34 H 38 N 10 O4[M+H]+651.3150, actual value is: 651.3160.

[0128] Example 2

[0129] (E)-N-(2-aminophenyl)-3-(4-(4-(2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-oxopiperazin-1-yl)phenyl)acrylamide (Compound Z2)

[0130] Step 1: Preparation of (E)-tert-butyl (2-(3-(4-(4-(2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-oxopiperazin-1-yl)methyl)phenyl)acrylamido)phenyl)carbamate (Intermediate 8)

[0131]

[0132] The intermediate 6 (0.24 mmol, 150 mg) was dissolved in DMF, and HATU (0.36 mmol, 136.8 mg) and DIEA (0.36 mmol, 46 mg) were added, and finally the raw material amine (0.24 mmol, 49.5 mg) was added. The mixture was stirred at room temperature and reacted for 30 min. The reaction progress was monitored by TLC. After the reaction was complete, a small amount of water was added to the reaction solution, and the mixture was extracted with EA, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 185 mg of the product with a yield of 93.4%.

[0133] Step 2: Preparation of compound Z2

[0134]

[0135] HCl dioxane (10 ml) was added to raw material A (150 mg, 0.18 mmol), and the mixture was stirred at room temperature for 2 h. The reaction progress was monitored by TLC. After the reaction was completed, the solid part was washed with DCM for three times, and then MeOH was added to dissolve the solid part. The product was concentrated under reduced pressure to obtain 112 mg of the product with a yield of 82%.

[0136] Characterization data: 1 H NMR (600MHz, DMSO-d6) δ11.02 (s, 1H), 8.23 ​​(dq, J = 6.9, 4.2Hz, 1H), 7.90 (dd ,J=6.1,3.1Hz,1H),7.83–7.59(m,8H),7.54(d,J=7.9Hz,1H),7.42(t,J=7.7 Hz,1H),7.31(t,J=7.6Hz,1H),7.12(d,J=15.8Hz,1H),5.41–4.70(m,7H),4. 49 (s, 6H), 4.33–3.83 (m, 10H), 3.51 (d, J = 7.4Hz, 2H), 1.47 (t, J = 7.4Hz, 3H).

[0137] HRMS (ESI+) m / z theoretical value: C 40 H 43 N 11 O3[M+H]+726.3623, actual value is: 726.3571.

[0138] Example 3

[0139] 6-(4-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-oxopiperazin-1-yl)-N-hydroxy-6-oxohexanamide (Compound Z3)

[0140] Step 1: Preparation of methyl 6-(4-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-oxopiperazin-1-yl)-6-oxohexanoate (Intermediate 9)

[0141]

[0142] The intermediate 4 (200 mg, 0.42 mmol) was dissolved in DMF (5 ml), and HATU (240 mg, 0.63 mmol) and DIEA (81 mg, 0.63 mmol) were added, followed by the raw material 6-methoxy-6-oxohexanoic acid (67 mg, 0.42 mmol). The mixture was stirred at room temperature for 30 min. After the reaction was complete, a small amount of water was added, and the mixture was extracted twice with EA, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 332 mg of a crude product. After purification by coating on a plate, 186 mg of the product was obtained, with a yield of 72%.

[0143] Step 2: Preparation of 6-(4-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-oxopiperazin-1-yl)-6-oxohexanoic acid (Intermediate 10)

[0144]

[0145] Dissolve NaOH (60 mg, 1.5 mmol) in water (2 ml) and methanol (4 ml), add intermediate 9 (186 mg, 0.3 mmol), stir at 70°C, react for 2 h, monitor the reaction progress by TLC, after the reaction is complete, adjust the pH to 5 with 1.5-2.5 N hydrochloric acid, spin dry the reaction solution and directly proceed to the next step.

[0146] Step 3: Preparation of 6-(4-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-oxopiperazin-1-yl)-6-oxo-N-((tetrahydro-2H-pyran-2-yl)oxy)hexanamide (Intermediate 11)

[0147]

[0148] The intermediate 10 (180 mg, 0.3 mmol) was dissolved in THF (4 ml), and HATU (170 mg, 0.45 mmol) and DIEA (58 mg, 0.45 mmol) were added and dissolved by ultrasonication. Then, the raw material O-(tetrahydro-2H-pyran-2-yl)hydroxylamine (35 mg, 0.3 mmol) was added and the reaction was stirred at room temperature for 30 min. The reaction progress was monitored by TLC. After the reaction was complete, a small amount of water was added, and the mixture was extracted twice with EA, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 192 mg of a crude product. The final product was purified by climbing a plate to obtain 115 mg with a yield of 55%.

[0149] Step 4: Preparation of compound Z3

[0150]

[0151] The intermediate 11 (115 mg, 0.16 mmol) was dissolved in methanol (4 ml), and 6N hydrochloric acid aqueous solution (0.2 ml) was added. The reaction was stirred at room temperature for 30 min and the reaction progress was monitored by TLC. After the reaction was complete, the pH was adjusted to 8 with sodium bicarbonate solution. The reaction liquid was spin-dried, filtered, and purified by climbing the plate to obtain 65 mg of the final product with a yield of 64%.

[0152] Characterization data: 1 H NMR(600MHz,DMSO-d6)δ10.37(s,1H),8.68(s,1H),8.10–7.94(m,1H),7.70 –7.59(m,1H),7.26(tt,J=7.3,5.6Hz,2H),4.88(s,2H),4.17(d,J=59.5Hz,6 H),3.79–3.67(m,8H),3.47(s,3H),3.26(q,J=7.4Hz,2H),2.34(dt,J=14.9, 6.9Hz, 2H), 1.96 (t, J = 7.0Hz, 2H), 1.55–1.44 (m, 4H), 1.33 (t, J = 7.4Hz, 3H).

[0153] HRMS (ESI+) m / z theoretical value: C 30 H 38 N 10 O5[M+H]+619.3099, actual value: 619.3083.

[0154] Example 4

[0155] 8-(4-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-oxopiperazin-1-yl)-N-hydroxy-8-oxooctylamide (Compound Z4)

[0156] Step 1: Preparation of methyl 6-(4-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-oxopiperazin-1-yl)-6-oxooctanoate (Intermediate 12)

[0157]

[0158] The intermediate 4 (200 mg, 0.42 mmol) was dissolved in DMF (5 ml), and HATU (240 mg, 0.63 mmol) and DIEA (81 mg, 0.63 mmol) were added, followed by the raw material 6-methoxy-6-oxooctanoic acid (80 mg, 0.42 mmol). The mixture was stirred at room temperature for 30 min. After the reaction was complete, a small amount of water was added, and the mixture was extracted twice with EA, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 340 mg of a crude product. After plate purification, 173 mg of the product was obtained with a yield of 64%.

[0159] Step 2: Preparation of 6-(4-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-oxopiperazin-1-yl)-6-oxooctanoic acid (Intermediate 13)

[0160]

[0161] NaOH (54 mg, 1.35 mmol) was dissolved in water (2 ml) and methanol (4 ml), and then intermediate 12 (173 mg, 0.27 mmol) was added. The mixture was stirred at 70°C for 2 h. The reaction progress was monitored by TLC. After the reaction was complete, the pH was adjusted to 5 with 1.5-2.5 N hydrochloric acid. The reaction solution was spin-dried and directly used for the next step.

[0162] Step 3: Preparation of 6-(4-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-oxopiperazin-1-yl)-6-oxo-N-((tetrahydro-2H-pyran-2-yl)oxy)octanamide (Intermediate 14)

[0163]

[0164] The intermediate 13 (170 mg, 0.27 mmol) was dissolved in THF (4 ml), and HATU (154 mg, 0.41 mmol) and DIEA (104 mg, 0.81 mmol) were added and dissolved by ultrasonication. Then, the raw material O-(tetrahydro-2H-pyran-2-yl)hydroxylamine (32 mg, 0.27 mmol) was added and the reaction was stirred at room temperature for 30 min. The reaction progress was monitored by TLC. After the reaction was complete, a small amount of water was added, and the mixture was extracted twice with EA, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 165 mg of a crude product. The final product was purified by climbing a plate to obtain 85 mg with a yield of 43%.

[0165] Step 4: Preparation of compound Z4

[0166]

[0167] The intermediate 14 (85 mg, 0.12 mmol) was dissolved in methanol (4 ml), and 6N hydrochloric acid aqueous solution (0.2 ml) was added. The reaction was stirred at room temperature for 30 min and the reaction progress was monitored by TLC. After the reaction was complete, the pH was adjusted to 8 with sodium bicarbonate solution. The reaction liquid was spin-dried, filtered, and purified by climbing the plate to obtain 36 mg of the final product with a yield of 46%.

[0168] Characterization data: 1 H NMR(600MHz,DMSO-d6)δ10.34(s,1H),8.66(s,1H),8.01(d,J=7.2Hz,1H),7.64 (d,J=7.1Hz,1H),7.26(q,J=6.5Hz,2H),4.88(s,2H),4.49–3.98(m,6H),3.84–3 .66(m,8H),3.51–3.39(m,3H),3.21(dd,J=45.2,6.1Hz,2H),2.33(dt,J=15.2,7 .3Hz,2H),1.93(t,J=7.6Hz,2H),1.48(q,J=8.5,7.8Hz,3H),1.39–1.18(m,8H).

[0169] HRMS (ESI+) m / z theoretical value: C 32 H 43 N 10 O5[M+H]+647.3412, actual value: 647.3400.

[0170] Example 5

[0171] 10-(4-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-oxopiperazin-1-yl)-N-hydroxy-10-oxodecanamide (Compound Z5)

[0172] Step 1: Preparation of methyl 6-(4-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-oxopiperazin-1-yl)-6-oxodecanoate (Intermediate 15)

[0173]

[0174] The intermediate 4 (200 mg, 0.42 mmol) was dissolved in DMF (5 ml), and HATU (240 mg, 0.63 mmol) and DIEA (81 mg, 0.63 mmol) were added, and then the raw material 6-methoxy-6-oxodecanoic acid (91 mg, 0.42 mmol) was added. The mixture was stirred at room temperature for 30 min. After the reaction was complete, a small amount of water was added, and the mixture was extracted twice with EA, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 280 mg of a crude product. After plate purification, 130 mg of the product was obtained, with a yield of 46%.

[0175] Step 2: Preparation of 6-(4-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-oxopiperazin-1-yl)-6-oxodecanoic acid (Intermediate 16)

[0176]

[0177] NaOH (40 mg, 0.97 mmol) was dissolved in water (2 ml) and methanol (4 ml), and then intermediate 15 (130 mg, 0.2 mmol) was added. The mixture was stirred at 70°C for 2 h. The reaction progress was monitored by TLC. After the reaction was complete, the pH was adjusted to 5 with 1.5-2.5 N hydrochloric acid. The reaction solution was spin-dried and directly used for the next step.

[0178] Step 3: Preparation of 6-(4-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-oxopiperazin-1-yl)-6-oxo-N-((tetrahydro-2H-pyran-2-yl)oxy)decanamide (Intermediate 17)

[0179]

[0180] The intermediate 16 (127 mg, 0.19 mmol) was dissolved in THF (4 ml), and HATU (110 mg, 0.29 mmol) and DIEA (74 mg, 0.58 mmol) were added and dissolved by ultrasonication. Then, raw material B (23 mg, 0.19 mmol) was added and the reaction was stirred at room temperature for 30 min. The reaction progress was monitored by TLC. After the reaction was complete, a small amount of water was added, and the mixture was extracted twice with EA, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by climbing a plate to obtain 122 mg of the final product with a yield of 84%.

[0181] Step 4: Preparation of compound Z5

[0182]

[0183] The intermediate 17 (122 mg, 0.16 mmol) was dissolved in methanol (8 ml), and a 6N hydrochloric acid aqueous solution (2 ml) was added. The mixture was stirred at room temperature for 30 min and the reaction progress was monitored by TLC. After the reaction was complete, the pH was adjusted to 8 with sodium bicarbonate solution. The reaction liquid was dried and filtered to obtain 102 mg of the product with a yield of 94%.

[0184] Characterization data: 1 H NMR(600MHz,DMSO-d6)δ10.36(d,J=6.0Hz,1H),8.66(s,1H),8.13–7.86(m,1H), 7.78–7.53(m,1H),7.36–7.16(m,2H),4.88(s,2H),4.57–3.96(m,6H),3.79–3.69 (m,8H),3.46(t,J=5.1Hz,1H),3.26(t,J=7.4Hz,2H),2.38–2.26(m,2H),1.93(t, J=7.3Hz,2H),1.50–1.43(m,4H),1.33(t,J=7.4Hz,3H),1.24(d,J=11.8Hz,10H).

[0185] Example 6

[0186] (E)-3-(4-(4-(2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)piperazin-1-yl)methylphenyl)-N-hydroxyacrylamide (Compound Z6)

[0187] Step 1: Preparation of tert-butyl 4-((2-chloro-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-piperazine-1-carboxylate (Intermediate 18)

[0188] Dissolve Boc piperazine (1.233 g, 6.619 mmol) in acetonitrile, potassium carbonate (2.740 g, 19.85 mmol), stir for 5 min, then add raw material 4-(2-chloro-8-(chloromethyl)-9-methyl-9H-purine-6-yl)morpholine (2 g, 6.619 mmol), stir and react for 4 hours at room temperature. Add water after sampling, extract with EA and then plate. After the raw material is basically reacted, add water and solid is found to precipitate, filter the reaction solution and keep the filter cake. Dissolve the filter cake with a mixed solvent of DCM and MeOH and dry it with anhydrous sodium sulfate, spin dry to obtain 1.29 g of product.

[0189] Step 2: Preparation of tert-butyl 4-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-piperazine-1-carboxylate (Intermediate 19)

[0190]

[0191] The intermediate 18 (1.500 g, 3.319 mmol) was dissolved in dioxane, and then the raw materials 2-ethyl-1H-benzo[d]imidazole (485 mg, 3.319 mmol), Pd2(dba)3 (343 mg, 0.332 mmol), XPhos (316 mg, 0.664 mmol), and Cs2CO3 (2.163 g, 6.638 mmol) were added in sequence. The mixture was refluxed at 110°C for 5 hours under nitrogen protection until the raw materials were completely converted. The reaction solution was extracted with EA water, and the upper organic solution was plated. It was observed that the raw materials were basically converted. EA, water extraction and column chromatography were performed to obtain 1.293 g of product with a yield of 69%.

[0192] Step 3: Preparation of 1-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)piperazine hydrochloride (Intermediate 20)

[0193]

[0194] Add intermediate 19 (2.708 g, 4.82 mmol) to a mixed solvent of HCl and 1,4-dioxane, and stir at room temperature for 20 minutes. The developing solvent is DCM:MeOH=5:1. The raw materials are basically reacted, and there is only one product point. Dichloromethane and methanol are added to separate the solid. After the reaction liquid is dried by spin drying, dichloromethane is added again to precipitate the solid. After removing the dichloromethane layer, spin drying is performed again to obtain 1.859 g of solid product, with a yield of 77%.

[0195] Step 4: Preparation of (E)-methyl 3-(4-(4-(2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-oxopiperazin-1-yl)methylphenyl)acrylate (Intermediate 21)

[0196]

[0197] The intermediate 20 (200 mg, 0.402 mmol) was dissolved in acetonitrile (5 ml), and then methyl 4-bromomethylcinnamate (102.45 mg, 0.402 mmol) and potassium carbonate (166.50 mg, 1.205 mmol) were added, and the mixture was stirred and reacted at 85°C under reflux. After sampling, DCM was added to the plate, and the developing solvent was DCM: MeOH = 10:1. The raw materials were basically reacted. The solvent was spin-dried, and the mixture was extracted with water and EA for 3 times. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and then spin-dried. After scraping, 100 mg of the product was obtained, and the yield was: 40%.

[0198] Step 5: Preparation of (E)-3-(4-(4-(2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-oxopiperazin-1-yl)methylphenyl)acrylic acid (Intermediate 22)

[0199]

[0200] Dissolve intermediate 21 (100 mg, 0.157 mmol) and LiOH monohydrate (19.77 mg, 0.472 mmol) in a mixed solvent (THF: MeOH: H2O = 4: 4: 1), and stir at 70 ° C for 2 hours. After sampling, add EA, and the developing solvent is DCM: MeOH = 10: 1. The raw materials are basically reacted. Spin dry acetonitrile and methanol until only water is left. Add hydrochloric acid to acidify under stirring. A white precipitate is precipitated and easily adheres to the wall of the bottle. Separate the solid product, add dichloromethane and methanol to dissolve it, spin it to half dry, and wash it with ethyl acetate. Filter by suction, and dry the filter cake to obtain a solid product, 80 mg, with a yield of 82%.

[0201] Step 6: Preparation of (E)-3-(4-((4-(2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-piperazin-1-yl)methyl)phenyl)-N-(((tetrahydro-2H-pyran-2-yl)oxy)acrylamide (Intermediate 23)

[0202]

[0203] The raw material O-(tetrahydro-2H-pyran-2-yl)hydroxylamine (18.84 mg, 0.16 mmol) was dissolved in DMF, and intermediate 22 (100 mg, 0.16 mmol), HATU (91.74 mg, 0.242 mmol), and DIEA (31.18 mg, 0.242 mmol) were added, and the reaction was carried out at room temperature for 3 hours. Samples were taken, and water and EA were added for extraction. The developing solvent was DCM: MeOH = 10:1. The raw materials were basically reacted. Water and EA were added for extraction. DMF should be removed as much as possible. The organic phases were combined, washed with saturated brine, dried with anhydrous sodium sulfate, and dried by rotation. After scraping, 46 mg of the product was obtained, and the yield was: 40%.

[0204] Step 7: Preparation of compound Z6

[0205]

[0206] The intermediate 23 (40 mg, 0.055 mmol) was dissolved in a 6 mol / L hydrochloric acid aqueous solution, reacted for half an hour, and then monitored by spotting. After sampling, DCM was added for extraction, and the plate was spotted. The developing solvent was DCM: MeOH = 5:1. The raw materials were basically reacted. Sodium hydroxide aqueous solution was added to adjust the base, and solids precipitated. The reaction solution was filtered, the filter cake was retained, and the solid was spin-dried to obtain 29.4 mg, with a yield of 83%.

[0207] Characterization structure: 1 H NMR (400MHz, DMSO-d6) δ8.02(d,J=6.9Hz,1H),7.63(d,J=6.6Hz,1H),7.46(dd,J=28.2,11.1Hz,3H),7.35–7.12(m,4H),6.49(d,J=15.5 Hz, 1H), 4.24 (s, 4H), 3.78 (dd, J = 13.3, 8.7Hz, 10H), 3.46 (s, 2H), 3.26 (q, J = 7.4Hz, 2H), 2.44 (d, J = 41.3Hz, 8H), 1.33 (t, J = 7.3Hz, 3H).

[0208] HRMS (ESI+) m / z theoretical value: C 34 H 40 N 10 O3[M+H]+637.3357, actual value is: 637.3346.

[0209] Example 7

[0210] 6-(4-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)piperazin-1-yl)-N-hydroxy-6-oxohexanamide (Compound Z8)

[0211] Step 1: Preparation of methyl 6-(4-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-piperazin-1-yl)-6-oxohexanoate (Intermediate 24)

[0212]

[0213] The raw material 6-methoxy-6-oxohexanoic acid (62.66 mg, 0.391 mmol) was dissolved in DMF, and HATU (223.14 mg, 0.587 mmol) and DIEA (75.84 mg, 0.587 mmol) were added in turn. After stirring for 5 minutes, the intermediate 20 (200 mg, 0.391 mmol) was added and reacted for about two hours. After sampling, EA and water were added for extraction, and the organic layer was spot-treated. The developing agent was DCM: MeOH = 10:1. The raw material was basically reacted. An appropriate amount of water and EA were added to the reaction solution for extraction, and the organic phase was collected three times. The spot plate was used to monitor the product content in the water and organic phase. Dry with anhydrous sodium sulfate, spin dry, and scrape to obtain 100 mg of the product, with a yield of 42%.

[0214] Step 2: Preparation of 6-(4-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-piperazin-1-yl)-6-oxohexanoic acid (Intermediate 25)

[0215]

[0216] Dissolve intermediate 24 (100 mg, 0.165 mmol) and LiOH monohydrate (20.29 mg, 0.495 mmol) in a mixed solvent (THF: MeOH: H2O = 4: 4: 1), and stir at 70 ° C for 2 hours. After sampling, add EA, and the developing solvent is DCM: MeOH = 10: 1. The raw materials are basically reacted. Spin dry acetonitrile and methanol until only water is left. Add hydrochloric acid to acidify under stirring. A white precipitate is precipitated and easily adheres to the wall of the bottle. Separate the solid product, add dichloromethane and methanol to dissolve it, spin it to half dry, and wash it with ethyl acetate. Filter by suction, and dry the filter cake to obtain a solid product, 70 mg, with a yield of 70%.

[0217] Step 3: Preparation of 6-(4-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-piperazin-1-yl)-6-oxo-N-((tetrahydro-2H-pyran-2-yl)oxy)hexanamide (Intermediate 26)

[0218]

[0219] The raw material O-(tetrahydro-2H-pyran-2-yl)hydroxylamine (13.59 mg, 0.116 mmol) was dissolved in DMF, and HATU (66.18 mg, 0.174 mmol) and DIEA (22.49 mg, 0.174 mmol) were added in turn. After stirring for 5 minutes, the intermediate 25 (70 mg, 0.116 mmol) was added and reacted for about two hours. After sampling, EA and water were added for extraction, and the organic layer was spot-treated. The developing agent was DCM:MeOH=10:1. The raw material was basically reacted. An appropriate amount of water and EA were added to the reaction solution for extraction, and the organic phase was collected three times. The spot plate was used to monitor the product content in the water and organic phase. Dry with anhydrous sodium sulfate, spin dry and scrape the product to obtain 46 mg, with a yield of 57%.

[0220] Step 4: Preparation of compound Z8

[0221]

[0222] The intermediate 26 (40 mg, 0.067 mmol) was dissolved in a 6 mol / L hydrochloric acid aqueous solution, reacted for half an hour, and then monitored by spotting. After sampling, DCM was added for extraction, and the plate was spotted. The developing solvent was DCM: MeOH = 5:1. The raw materials were basically reacted. Sodium hydroxide aqueous solution was added to adjust the base, and solids precipitated. The reaction solution was filtered, the filter cake was retained, and the solid was spin-dried to obtain 30 mg, with a yield of 75%.

[0223] Characterization structure: 1 H NMR(600MHz,DMSO-d6)δ10.40(d,J=18.3Hz,1H),8.70(s,1H),8.02(d,J=7.6Hz,1H), 7.84–7.58(m,1H),7.26(q,J=6.8,6.2Hz,2H),4.53–4.00(m,3H),3.95–3.67(m,7H), 3.46(s,8H),3.27(q,J=7.4Hz,2H),2.44(dt,J=28.2,5.0Hz,3H),2.29(t,J=7.1Hz,2 H), 1.96 (t, J = 6.9Hz, 2H), 1.47 (ddt, J = 30.3, 15.6, 7.4Hz, 4H), 1.34 (t, J = 7.4Hz, 3H).

[0224] Example 8

[0225] 8-(4-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)piperazin-1-yl)-N-hydroxy-8-oxooctylamide (Compound Z9)

[0226] Step 1: Preparation of methyl 6-(4-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-piperazin-1-yl)-6-oxooctanoate (Intermediate 27)

[0227]

[0228] The raw material 6-methoxy-6-oxooctanoic acid (132.40 mg, 0.704 mmol) was dissolved in DMF, and then HATU (401.2 mg, 1.06 mmol) and DIEA (135.2 mg, 1.06 mmol) were added in sequence. After stirring for 5 minutes, the intermediate 20 (350 mg, 0.704 mmol) was added and reacted for about two hours. After sampling, EA and water were added for extraction, and the organic layer was spot-treated. The developing agent was DCM: MeOH = 30:1. The raw material was basically reacted. An appropriate amount of water and EA were added to the reaction solution for extraction, and the organic phase was collected three times. The spot plate was used to monitor the product content in the water and organic phase. Dry with anhydrous sodium sulfate, spin dry, and scrape to obtain 215 mg of the product, with a yield of 48%.

[0229] Step 2: Preparation of 6-(4-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-piperazin-1-yl)-6-oxooctanoic acid (Intermediate 28)

[0230]

[0231] Intermediate 27 (215 mg, 0.398 mmol) and NaOH (79.53 mg, 1.988 mmol) were dissolved in a mixed solvent (MeOH:H2O=2:1) ​​and stirred at 70°C for 2 hours. EA was added after sampling, and the developing solvent was DCM:MeOH=10:1. The raw materials were basically reacted. Acetonitrile and methanol were dried by spin drying until only water was left. Hydrochloric acid was added under stirring to adjust the pH to 5-6. The reaction liquid was dried by spin drying and used directly in the next step. The amount of other raw materials in the next step was calculated according to the theoretical yield.

[0232] Step 3: Preparation of 6-(4-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-piperazin-1-yl)-6-oxo-N-((tetrahydro-2H-pyran-2-yl)oxy)octanamide (Intermediate 29)

[0233]

[0234] The raw material O-(tetrahydro-2H-pyran-2-yl)hydroxylamine (46.63 mg, 0.398 mmol) was dissolved in DMF, and HATU (227.00 mg, 0.597 mmol) and DIEA (154.31 mg, 0.597 mmol) were added in turn. After stirring for 5 min, intermediate 28 (245.70 mg, 0.398 mmol) was added and reacted for about two hours. After sampling, EA and water were added for extraction, and the organic layer was spot-treated. The developing agent was DCM:MeOH=10:1. The raw material was basically reacted. An appropriate amount of water and EA were added to the reaction solution for extraction, and the organic phase was collected three times. The spot plate was used to monitor the product content in the water and organic phase. Dry with anhydrous sodium sulfate, spin dry and scrape the product to obtain 130 mg, with a yield of 57%.

[0235] Step 4: Preparation of compound Z9

[0236]

[0237] Intermediate 29 (65 mg, 0.067 mmol) was dissolved in 6 mol / L hydrochloric acid aqueous solution, reacted for half an hour, and then monitored by spot plate. After sampling, DCM was added for extraction, and the developing solvent was DCM: MeOH = 5:1. The raw materials were basically reacted. Sodium hydroxide aqueous solution was added to adjust the base, and solids precipitated. The reaction solution was filtered, the filter cake was retained, and the solid was spin-dried to obtain 30 mg, with a yield of 63%.

[0238] Characterization structure: 1H NMR(600MHz,DMSO-d6)δ10.37(s,1H),8.69(s,1H),8.19–7.90(m,1H),7.73–7.55(m,1H) ),7.26(dt,J=11.9,7.4Hz,2H),4.52–4.05(m,3H),3.90–3.71(m,7H),3.44(d,J=12.8Hz ,8H),3.27(q,J=7.4Hz,2H),2.44(dt,J=26.7,4.9Hz,3H),2.27(t,J=7.5Hz,2H),1.94( t,J=7.4Hz,2H),1.46(tt,J=12.7,6.3Hz,4H),1.34(t,J=7.4Hz,3H),1.30–1.18(m,4H).

[0239] Example 9

[0240] 10-(4-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)piperazin-1-yl)-N-hydroxy-10-oxodecanamide (Compound Z10)

[0241] Step 1: Preparation of methyl 6-(4-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-piperazin-1-yl)-6-oxodecanoate (Intermediate 30)

[0242]

[0243] The raw material 6-methoxy-6-oxodecanoic acid (84.51 mg, 0.391 mmol) was dissolved in DMF, and then HATU (223.14 mg, 0.587 mmol) and DIEA (75.84 mg, 0.587 mmol) were added in sequence. After stirring for 5 minutes, the intermediate 20 (200 mg, 0.391 mmol) was added and reacted for about two hours. After sampling, EA and water were added for extraction, and the organic layer was spot-treated. The developing agent was DCM: MeOH = 30:1. The raw material was basically reacted. An appropriate amount of water and EA were added to the reaction solution for extraction, and the organic phase was collected three times. The spot plate was used to monitor the product content in the water and organic phase. Dry with anhydrous sodium sulfate, spin dry, and scrape to obtain 115 mg of the product, with a yield of 40%.

[0244] Step 2: Preparation of 6-(4-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-piperazin-1-yl)-6-oxodecanoic acid (Intermediate 31)

[0245]

[0246] Intermediate 30 (115 mg, 0.174 mmol) and NaOH (34.8 mg, 0.87 mmol) were dissolved in a mixed solvent (MeOH:H2O=2:1) ​​and stirred at 70°C for 2 hours. EA was added after sampling, and the developing solvent was DCM:MeOH=10:1. The raw materials were basically reacted. Acetonitrile and methanol were dried by spin drying until only water was left. Hydrochloric acid was added under stirring to adjust the pH to 5-6. The reaction liquid was dried by spin drying and used directly in the next step. The amount of other raw materials in the next step was calculated according to the theoretical yield.

[0247] Step 3: Preparation of 6-(4-((2-(2-ethyl-1H-benzo[d]imidazol-1-yl)-9-methyl-6-morpholino-9H-purin-8-yl)methyl)-3-piperazin-1-yl)-6-oxo-N-((tetrahydro-2H-pyran-2-yl)oxy)decanamide (Intermediate 32)

[0248]

[0249] The raw material O-(tetrahydro-2H-pyran-2-yl)hydroxylamine (20.38 mg, 0.174 mmol) was dissolved in DMF, and HATU (99.24 mg, 0.261 mmol) and DIEA (33.73 mg, 0.261 mmol) were added in turn. After stirring for 5 minutes, the intermediate 31 (115 mg, 0.174 mmol) was added and reacted for about two hours. After sampling, EA and water were added for extraction, and the organic layer was spot-treated. The developing agent was DCM:MeOH=10:1. The raw material was basically reacted. An appropriate amount of water and EA were added to the reaction solution for extraction, and the organic phase was collected three times. The spot plate was used to monitor the product content in the water and organic phase. Dry with anhydrous sodium sulfate, spin dry and scrape the product to obtain 50 mg, with a yield of 35%.

[0250] Step 3: Preparation of compound Z10

[0251]

[0252] Intermediate 32 (50 mg, 0.067 mmol) was dissolved in 6 mol / L hydrochloric acid aqueous solution, reacted for half an hour, and then monitored by spotting. After sampling, DCM was added for extraction, and the developing solvent was DCM: MeOH = 5:1. The raw materials were basically reacted. Sodium hydroxide aqueous solution was added to adjust the base, and solids precipitated. The reaction solution was filtered, the filter cake was retained, and the solid was spin-dried to obtain 25 mg, with a yield of 50%.

[0253] Characterization structure: 1H NMR(600MHz,DMSO-d6)δ10.32(s,1H),8.65(s,1H),8.15–7.97(m,1H),7.69–7.56(m,1H),7. 26(h,J=7.7,7.0Hz,2H),4.44–4.10(m,3H),3.87–3.76(m,7H),3.44(dd,J=11.0,5.8Hz,8H) ,3.27(d,J=7.4Hz,2H),2.44(dt,J=26.5,5.0Hz,3H),2.27(q,J=8.9,8.2Hz,2H),1.93(t,J= 7.4Hz, 2H), 1.47 (q, J = 12.2, 9.7Hz, 4H), 1.34 (t, J = 7.4Hz, 3H), 1.24 (dd, J = 9.6, 4.9Hz, 8H).

[0254] The beneficial effects of the present invention are demonstrated below through specific experimental examples.

[0255] Experimental Example 1: Affinity test of compound and target protein

[0256] The inhibition rate and IC of the compounds on PI3K family protein PI3Kδ and HDAC family protein HDAC6 50 The determination was provided by Wise Chemical Research Co., Ltd. The positive control compound PI103 and the positive control compound SAHA were used as controls.

[0257] 1.1 The inhibition rate of the compound on PI3Kδ was detected by ADP-Glo ​​Luminescent Assay. The specific experimental method is as follows:

[0258] 1.1.1 Compound preparation

[0259] 1) Compound dilution

[0260] The initial concentration of the compound tested on the kinase is 1 μM, and it needs to be prepared to a 100-fold concentration, i.e. 100 μM.

[0261] Add 99 μL of 100% DMSO to the second well of the 384-well Echo plate, and then add 1 μL of 10 mM compound solution to prepare a 100 μM compound solution. Add 30 μL of 100% DMSO to the other wells. Take 15 μL of compound from the second well and add it to the third well, and then make 3-fold dilutions down, for a total of 8 concentrations. Dilution instrument: automatic microporous pipette (PrecisionPRC384U). Transfer 40 μL of 100% DMSO to two empty wells as Max well and Min well.

[0262] 2) Transfer compounds to the reaction plate

[0263] Using an Echo 650, 50 nl of compound was transferred into a 384-well assay plate.

[0264] 1.1.2 Preparation of 1x kinase buffer

[0265] PI3Kδ 1x Kinase Buffer

[0266] 50 mM HEPES, pH 7.5

[0267] 3mM MgCl2

[0268] 1mM EGTA

[0269] 100mM NaCl

[0270] 0.03% CHAPS

[0271] 2mM DTT

[0272] 1.1.3 Kinase reaction and termination

[0273] 1) Add kinase to 1x kinase buffer to form a 2x kinase solution;

[0274] 2) Transfer 2.5 μL of the above 2x kinase solution to the reaction wells of the 384-well plate, and add 1x kinase buffer to the negative control wells. Incubate at room temperature for 10 minutes;

[0275] 3) Add substrate and ATP and other reagents to 1x kinase buffer to form 2x substrate solution;

[0276] 4) Transfer 2.5 μL of 2x substrate solution to the reaction wells of a 384-well plate;

[0277] 5) Incubate at room temperature for 60 minutes;

[0278] 6) Transfer 5 μL ADP-Glo ​​reagent 1 to the 384-well plate to terminate the reaction;

[0279] 7) react at room temperature for 120 minutes;

[0280] 8) Transfer 10 μL ADP-Glo ​​reagent 2 to a 384-well plate;

[0281] 9) Leave at room temperature for 30 minutes.

[0282] 1.1.4 Data Reading

[0283] The RLU values ​​were read on Envision2104 Multilable Reader.

[0284] 1.1.5 Data Calculation

[0285] 1) Copy the RLU reading

[0286] 2) Convert the above data into inhibition percentage using the formula

[0287] Percent inhibition=(max-sample RLU) / (max-min)*100

[0288] "min" is the reading of the control well without enzyme reaction; "max" is the reading of the control well with DMSO added

[0289] 3) Import the data into MS Excel and use XLFit excel add-in version 5.4.0.8 to perform curve fitting. Fitting formula: Y = Bottom + (Top-Bottom) / (1 + (IC50 / X)^HillSlope)

[0290] 1.2 The inhibition rate of the compound on HDAC6 was detected by Fluroscence assay. The specific experimental method is as follows:

[0291] 1.2.1 Prepare 1x buffer

[0292] Prepare 1x assay buffer (modified Tris buffer).

[0293] 1.2.2 Compound serial dilution:

[0294] Compounds were transferred to assay plates by Echo and diluted three-fold in 100% DMSO.

[0295] 1.2.3 Prepare enzyme solution in 1x assay buffer.

[0296] 1.2.4 Prepare the matrix solution:

[0297] Make substrate solution by adding trypsin and Ac-peptide substrate in 1x assay buffer.

[0298] 1.2.5 Transfer 15 μL of enzyme solution to the assay plate, or 15 μL of 1x assay buffer for low control, and incubate at room temperature for 15 minutes.

[0299] 1.2.6 Add 10 μL of substrate solution to each well to start the reaction.

[0300] 1.2.7 Kinetically read the plate on Envision with an excitation wavelength of 355 nm and an emission wavelength of 460 nm.

[0301] 1.2.8 Curve Fitting

[0302] Fit the data in Excel using equation (1) to obtain inhibition values

[0303] Equation (1): Inh% = (maximum signal) / (maximum minimum) * 100

[0304] The data were fitted in XL Fit using equation (2) to obtain IC50 values

[0305] Equation (2): Y = bottom + (top - bottom) / (1 + (IC50 / X) * hillside)

[0306] Y is the percentage of inhibition and X is the compound concentration.

[0307] The results of the inhibitory effects of the compounds of the present invention on PI3Kδ and HDAC6 activities are shown in Table 1 below. 50 Value ≤ 5nM; letter B means 5nM ≤ IC 50 Value ≤20nM; letter C means 20nM≤IC 50 Value ≤ 100nM; letter C means 100nM ≤ IC 50 Value ≤1000nM.

[0308] Table 1 Statistical table of the inhibitory effects of the compounds in the examples on PI3Kδ and HDAC6 activity

[0309]

[0310]

[0311] As can be seen from Table 1, the inhibitory activity of the compounds of the present invention is comparable to or better than the positive control compound PI103 (a known PI3Kδ single-target inhibitor). The inhibitory activity of most compounds of the present invention is comparable to the positive control compound SAHA (a known HDAC6 single-target inhibitor) for HDAC6.

[0312] More importantly, most of the compounds of the present invention showed inhibitory effects on PI3Kδ and HDAC6 at nanomolar concentrations. Among them, the preferred compounds Z1, Z5, and Z9 have an IC of 50 The value is ≤5nM, and the IC 50 The value is between 5 nM and 20 nM. This indicates that the compound of the present invention can be used to prepare a dual-target inhibitor that simultaneously inhibits PI3Kδ and HDAC6.

[0313] Experimental Example 2: Cell proliferation inhibitory activity test of compounds

[0314] The half inhibitory concentration (IC50) of the compounds on various tumor cells was investigated by MTS method and Cell Titer-Glo cell viability assay. 50 ).

[0315] The tumor cells used in the experiment are: Jeko-1 and CA-46 cells.

[0316] Experimental method: Collect cells in the logarithmic growth phase in a centrifuge tube, place the centrifuge tube in a centrifuge, set to 1200r / min, centrifuge for 3min. Resuspend the cells with fresh culture medium, count the resuspended cells, and dilute to a cell concentration of 8×10 4 ~1×10 5 / mL cell suspension, inoculate cells into 96-well plates according to 100μL cell suspension per well (i.e. 8000-10000 cells per well), and place the inoculated 96-well plates in an incubator for incubation. Dilute the drug gradient to an appropriate concentration (generally three times dilution), treat the inoculated cells with 100μL per well, set up 3 replicates for each drug concentration, and treat the cells with fresh culture medium containing 0.1% DMSO as a negative control group. Place the 96-well plate in the incubator and continue to culture for the required time (24h, 48h, 72h, 96h, 120h), then take it out, add 20μL of MTS solution to each well, incubate at 37℃ for 2-4 hours (2h is most suitable), and use an enzyme marker to detect the absorbance value at a wavelength of 492nm after the test solution and the color developer generate a colored solution. The absorbance value (A) is proportional to the number of living cells. The inhibitory rate of the drug on tumor cells was calculated according to the following formula: cell growth inhibition rate = (control group A495nm - experimental group A495nm) / control group A495nm × 100%. The relationship between cell growth inhibition rate and drug concentration was calculated using Graphpad Prism software, and the drug growth inhibition curve was fitted and the half inhibitory concentration (IC 50 )value.

[0317] The proliferation inhibition activities of the compounds of the present invention on non-Hodgkin's lymphoma cell lines CA-46 and JEKO-1 are shown in Table 2. Wherein, the letter A represents IC 50 Value ≤50nM; letter B means 50nM≤IC 50 Value ≤ 500nM; letter C indicates 500μM ≤ IC 50 Value ≤ 5 μM; letter D represents IC 50 Values ​​> 5 μM.

[0318] Table 2. Statistical table of the inhibitory activity of the compounds of the present invention on tumor cell proliferation

[0319]

[0320] According to the results in Table 2, the compounds of the present invention can effectively inhibit the proliferation of lymphoma cell lines, and the IC 50 The value can reach nanomolar level.

[0321] In summary, the present invention provides a heterocyclic compound, which has good inhibitory activity against both PI3K and HDAC targets, and can be used as a dual-target inhibitor with excellent effects. The compound of the present invention can be used to prepare drugs for preventing and / or treating diseases related to PI3K and / or HDAC, and has good application prospects.

Claims

1. The compound represented by formula I, its salt, its stereoisomer or its solvate: in, R1 is LR a R b CONHR c , L is selected from none, C1-C6 alkylene, carbonyl; R a Select from None, C1~C 10 Alkylene, 6- to 10-membered aryl which is unsubstituted or substituted by one or more R', 5- to 10-membered heteroaryl which is unsubstituted or substituted by one or more R'; R b Select from None, C2~C 10 alkenylene; R c is selected from hydroxyl, 6-10 membered aryl which is unsubstituted or substituted by one or more R'; And L, R a , R b Not at the same time is nothing; R2, R3, R4, R5, R6, R7, R8, and R9 are independently selected from hydrogen, or two substituents on the same carbon atom form =O, and at most one of R2, R3, R4, R5, R6, R7, R8, and R9 forms =O; R 10 Selected from C1-C6 alkyl groups which are unsubstituted or substituted by one or more R'; R 11 is selected from a cyclic ring which is unsubstituted or substituted with one or more R'; R' is independently selected from hydroxy, halogen, cyano, amino, C1-C6 alkyl, C2-C6 alkenyl, C1-C6 alkoxy, and C2-C6 alkynyl.

2. The compound, salt thereof, stereoisomer thereof or solvate thereof according to claim 1, characterized in that: The compound is shown in Formula II: in, R1' is selected from C1-C6 alkyl; R1 is LR a R b CONHR c , L is selected from none, C1-C6 alkylene, carbonyl; R a Select from None, C1~C 10 Alkylene, 6- to 10-membered aryl which is unsubstituted or substituted by one or more R', 5- to 10-membered heteroaryl which is unsubstituted or substituted by one or more R'; R b Select from None, C2~C 10 alkenylene; R c is selected from hydroxyl, 6-10 membered aryl which is unsubstituted or substituted by one or more R'; And L, R a , R b Not at the same time is nothing; R2, R3, R4, R5, R6, R7, R8, and R9 are independently selected from hydrogen, or two substituents on the same carbon atom form =O, and at most one of R2, R3, R4, R5, R6, R7, R8, and R9 forms =O; R 10 Selected from C1-C6 alkyl groups which are unsubstituted or substituted by one or more R'; R' is independently selected from hydroxy, halogen, cyano, amino, C1-C6 alkyl, C2-C6 alkenyl, C1-C6 alkoxy, and C2-C6 alkynyl.

3. The compound, salt thereof, stereoisomer thereof or solvate thereof according to claim 2, characterized in that: R1' is selected from C1-C6 alkyl; Selected from R1 is LR a R b CONHR c , L is selected from none, C1-C6 alkylene, carbonyl; R a Select from None, C1~C 10 Alkylene, 6- to 10-membered aryl which is unsubstituted or substituted by one or more R', 5- to 10-membered heteroaryl which is unsubstituted or substituted by one or more R'; R b Select from None, C2~C 10 alkenylene; R c is selected from hydroxyl, 6-10 membered aryl which is unsubstituted or substituted by one or more R'; And L, R a , R b Not at the same time is nothing; R 10 Selected from C1-C6 alkyl groups which are unsubstituted or substituted by one or more R'; R' is independently selected from amino, C1-C6 alkyl, and C2-C6 alkenyl.

4. The compound, salt thereof, stereoisomer thereof or solvate thereof according to claim 3, characterized in that: The compound is shown in Formula III, Formula IV, and Formula V: in, R1' is selected from C1-C3 alkyl; R 10 Selected from C1-C3 alkyl; R1 is LR a R b CONHR c , L is selected from none, C1-C3 alkylene, carbonyl; R a Selected from none, C1-C8 alkylene, phenyl; R b Selected from none, C2-C3 alkenylene; And L, R a , R b Not at the same time is nothing; R c is selected from hydroxyl, phenyl which is unsubstituted or substituted by one or more R'; R' is each independently selected from amino groups; Preferably, R1 is 5. The compound, salt, stereoisomer or solvate thereof according to any one of claims 1 to 4, characterized in that: The compound is one of the following compounds:

6. Use of the compound according to any one of claims 1 to 5, its salt, its stereoisomer or its solvate in the preparation of a PI3K inhibitor and / or a HDAC inhibitor.

7. The use according to claim 6, characterized in that: The PI3K inhibitor is a PI3Kδ inhibitor, and the HDAC inhibitor is an HDAC6 inhibitor.

8. Use of the compound according to any one of claims 1 to 5, its salt, its stereoisomer or its solvate in the preparation of a medicament for preventing and / or treating diseases associated with PI3K and / or HDAC.

9. The use according to claim 8, characterized in that: The disease associated with PI3K and / or HDAC is cancer, and the cancer is preferably lymphoma.

10. A pharmaceutical composition, characterized in that: The invention relates to a preparation prepared by taking the compound, salt, stereoisomer or solvate thereof as any one of claims 1 to 5 as active ingredient and adding pharmaceutically acceptable auxiliary materials or supplementary ingredients.

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