A dual-targeting small molecule drug and use thereof

Abstract

The application discloses a compound shown in formula I, or a pharmaceutically acceptable salt, isomer, metabolite, prodrug, solvate or hydrate thereof, a pharmaceutical composition and use. The deuterated isoquinoline-amino pyrimidine compound shown in formula I provided by the application has obvious effects on blocking VISTA and PD-1 / PD-L1 signal pathways, can effectively treat and relieve diseases such as cancers, and is simple in preparation as a small molecule inhibitor and convenient for industrial production.

Description

Technical Field

[0001] This invention relates to the field of chemical and pharmaceutical technology, specifically to compounds that block the VISTA and PD-1 / PD-L1 signaling pathways, their preparation methods, and uses. Background Technology

[0002] Malignant tumors are among the most complex and difficult-to-treat diseases in the world today, seriously threatening human health and life. Tumor treatment primarily includes surgery, radiotherapy, chemotherapy, and targeted therapy. Tumor immunotherapy refers to a treatment method that stimulates the body's immune system to enhance anti-tumor immune responses, thereby inhibiting and killing tumor cells. With the integrated development and cross-fertilization of oncology, immunology, and molecular biology, and in-depth research into the pathogenesis of tumors, immunotherapy has achieved multifaceted results, bringing new hope to tumor treatment.

[0003] Immune checkpoint inhibitors are currently a popular type of immunotherapy drug in tumor immunotherapy. Tumor cells achieve immune escape by upregulating the expression of immune checkpoint receptors and inhibiting the activity of immune T cells. Immune checkpoint inhibitors work by inhibiting immune checkpoint pathways, relieving the inhibition of immune T cells, activating the body's immune killing of tumor cells, and thus achieving the effect of tumor treatment. Currently identified immune checkpoints include CTLA-4 (cytotoxic T lymphocyte-associated antigen-4), PD-1 (Programmed cell death 1), and TIM3 (T cell membrane 3) (see Drew M. Pardoll, Nature Review Cancer, 2012, 12, 252).

[0004] Programmed death receptor 1 (PD-1) is a type I transmembrane protein composed of 288 amino acids. It consists of an immunoglobulin (Ig) superfamily domain, a stalk of approximately 20 amino acids, a transmembrane domain, and an intracellular domain of approximately 95 amino acid residues, which contains the immunoreceptor tyrosine motif inhibitory motif (ITIM) and the immunoreceptor tyrosine switch motif (ITSM). PD-1 is primarily expressed on T cells, B cells, natural killer T cells, activated monocytes, and dendritic cells. PD-1 has two natural ligands: PD-L1 and PD-L2, which can interact with PD-1 to transmit inhibitory signals and regulate the balance between T cell activation, tolerance, and immunopathology. However, tumor cells can also express PD-L1, which interacts with PD-1 on T cells, thereby suppressing T cell immune activity and causing immune escape. Therefore, inhibiting the interaction between PD-1 and PD-L1 can activate the body's anti-tumor immune response, thereby achieving the effect of killing tumor cells. Several monoclonal antibody drugs targeting the PD-1 / PD-L1 signaling pathway have been approved for marketing and use in the treatment of cancer patients. In addition, small molecule inhibitors of PD-1 / PD-L1 are under active development, with the most advanced reaching Phase II clinical trials. However, as clinical treatment of cancer patients progresses, studies have found that inhibition of the PD-1 / PD-L1 pathway can promote increased expression of other immune checkpoints, such as VISTA and TIM3, leading to drug tolerance.

[0005] The immunoglobulin variable domain (VISTA), a T-cell activation inhibitor, is an immune checkpoint primarily expressed in hematopoietic tissues. It is also highly expressed in bone marrow cells, neuronal cells, and neutrophils. Unlike other immune checkpoints that are induced to express after immune response activation, VISTA is stably expressed during immune cell homeostasis. Multiple studies have shown that VISTA also has an inhibitory effect on the immune system. Therefore, inhibiting the VISTA signaling pathway can also restore the body's anti-tumor immune activity.

[0006] To better leverage the body's anti-tumor immune response, research on dual-target small molecule inhibitors is an effective approach. Currently, there are no commercially available small molecule inhibitors that simultaneously target the PD1 / PD-L1 and VISTA signaling pathways. Therefore, developing novel dual-target small molecule inhibitors with good anti-tumor activity is of great significance. Summary of the Invention

[0007] Purpose of the invention: In view of the current situation that there are no dual-target inhibitors of the VISTA and PD-1 / PD-L1 signaling pathways on the market, this invention provides a small molecule compound that simultaneously targets the VISTA and PD-1 / PD-L1 signaling pathways, as well as its preparation method and uses.

[0008] Technical solution: To achieve the above objectives, this invention discloses a compound represented by Formula I, and its pharmaceutically acceptable salt:

[0009]

[0010] Wherein, X1 is independently CH2, O, NH or S;

[0011] R1 is independently an aldehyde, hydroxyl, substituted or unsubstituted amino group, or an amino acid;

[0012] R2 is independently hydrogen, deuterium, halogen, cyano, morpholino, tetrahydropyrano, substituted or unsubstituted hydroxyl, substituted or unsubstituted amino, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkoxy, substituted or unsubstituted cycloalkyl, substituted or unsubstituted pyrrolidone, substituted or unsubstituted piperidinyl, substituted or unsubstituted piperazine.

[0013] R3 and R4 are each hydrogen, deuterium, halogen, cyano, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted alkoxy.

[0014] m can be 0, 1, 2, or 3;

[0015] n can be 0, 1, 2, 3, 4, or 5.

[0016] Furthermore, the present invention discloses a compound represented by Formula I, wherein in each R1, the substituent in the substituted amino group is one or more of the following groups: C 1-4 Alkyl, C 1-4 amide group, C 1-4 Ester group, C 14 Carboxyl group, C 1-4 hydroxyl group; wherein the C 1-4 Alkyl, C 1-4 amide group, C 1-4 Ester group, C 1-4 Carboxyl group, C 1-4 The hydroxyl group may optionally be substituted with one or more of the following substituents: hydroxyl, carboxyl, cyano, amino, C 3-6 cycloalkyl, C 6-10 Aryl, C 6-10 Heterocyclic group, C 2-4 alkenyl, C 2-4Alkyne group; when there are multiple substituents, the substituents may be the same or different.

[0017] Furthermore, the present invention discloses a compound represented by Formula I, wherein in each R2, the substituents in the substituted hydroxyl, substituted amino, substituted alkyl, substituted alkenyl, substituted alkynyl, substituted alkoxy, substituted C3-C6 cycloalkyl, substituted pyrrolidone, substituted piperidinyl, and substituted piperazine groups can be one or more of the following groups: halogen, cyano, hydroxyl, trifluoromethyl, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Carboxyl group, C 1-4 ester group or C 1-4 amide group; wherein the C 1-4 Alkyl, C 1-4 amide group, C 1-4 Ester group, C 1-4 Carboxyl group, C 1-4 The hydroxyl group may optionally be substituted with one or more of the following substituents: hydroxyl, carboxyl, cyano, amino, C 3-6 cycloalkyl, C 6-10 Aryl, C 6-10 Heterocyclic group, C 2-4 alkenyl, C 2-4 Alkyne group; when there are multiple substituents, the substituents may be the same or different.

[0018] The compound shown in Formula I of this application is specifically selected from the following compounds:

[0019]

[0020]

[0021] Furthermore, the specific compounds mentioned above also include their pharmaceutically acceptable salts, racemates, optical isomers, or solvent compounds.

[0022] The synthetic route for preparing the above-mentioned compounds, when X is O, NH, S, R1 is a substituted or unsubstituted amino group, and m is 1, is as follows:

[0023]

[0024] Where R2, R3, R4, and n are consistent with the definitions in Equation I, and the synthesis steps are as follows:

[0025] (1) Compound II and Compound III undergo a condensation cyclization reaction to give Compound IV;

[0026] (2) Compound IV reacts with pinacol diboronic acid ester to give compound V;

[0027] (3) Compound VI was given by a nucleophilic substitution reaction to give compound VII;

[0028] (4) Compound V and compound VII undergo a Suzuki coupling reaction to give compound VIII;

[0029] (5) Compound VIII was reduced to obtain compound IX.

[0030] (6) Compound IX is oxidized to obtain compound X.

[0031] (7) Compound X was subjected to a reducing amination reaction to obtain compound XI.

[0032] A pharmaceutical composition comprising a therapeutically effective amount of one or more compounds having the structure of general formula I or a pharmaceutically acceptable salt, racemate, optical isomer or solvent compound thereof as an active ingredient and a pharmaceutically acceptable carrier.

[0033] The pharmaceutical composition is a capsule, powder, tablet, granule, pill, injection, syrup, oral liquid, inhaler, ointment, suppository or patch.

[0034] Use of compounds with the structure described in Formula I in the preparation of immune checkpoint inhibitors, inhibitors with VISTA and PD-1 / PD-L1 signaling pathway inhibitory activities, antitumor drugs, and anti-infective drugs.

[0035] Use of the pharmaceutical composition in the preparation of immune checkpoint inhibitors, inhibitors with VISTA and PD-1 / PD-L1 signaling pathway inhibitory activities, antitumor drugs, and anti-infective drugs.

[0036] Beneficial effects: The compounds of this invention have novel structures, can be administered orally, and have significant effects on blocking the VISTA and PD-1 / PD-L1 signaling pathways. They can effectively treat and alleviate diseases such as cancer, and are simple to prepare as small molecule inhibitors, which is convenient for industrial production. Detailed Implementation

[0037] The technical solution of the present invention will be clearly and completely described below with reference to specific embodiments. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention. In addition, it is worth noting that the raw materials involved in the present invention are all commercially available products unless otherwise specified.

[0038] Example 1: Preparation of Compound 1

[0039]

[0040] Synthesis route:

[0041]

[0042] Synthesis of compound I-2

[0043] Starting material I-1 (5 g, 26.59 mmol) and methyl 3-formaldehyde benzoate (4.54 g, 27.66 mmol) were dissolved in anhydrous ethanol (40 mL) and refluxed in an oil bath at 80 °C for 3 h. The reaction solution was then cooled to room temperature, the solvent was concentrated to dryness, and then dissolved in anhydrous dichloromethane (40 mL). 6.04 g of 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) was added, and the reaction was carried out at room temperature for 1 h under nitrogen protection. The reaction was confirmed by TLC to be complete. The mixture was extracted with ethyl acetate, washed with saturated sodium thiosulfate solution and saturated sodium bicarbonate solution, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to prepare slurry. The slurry was purified by column chromatography (petroleum ether:ethyl acetate = 20:1) to give compound 2 (7.2 g, 82%).

[0044] Synthesis of compound I-3

[0045] Compound I-2 (2 g, 6.02 mmol), pinacol diborate (2.29 g, 9.03 mmol), 1,1′-bis(diphenylphosphine)ferrocene palladium dichloride (220 mg, 0.301 mmol), and potassium acetate (1.77 g, 18.06 mmol) were dissolved in 1,4-dioxane (20 mL), protected with a nitrogen balloon, and stirred in an oil bath at 80 °C for 12 h. TLC analysis showed that the reaction proceeds were complete. The mixture was filtered through diatomaceous earth, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (petroleum ether:ethyl acetate = 20:1) to give compound 3 (1.5 g, 71%).

[0046] Synthesis of compound I-4

[0047] m-Bromophenol (2 g, 11.56 mmol) was dissolved in acetonitrile (20 mL), and 1-bromo-3-chloropropane (2.18 g, 13.87 mmol) and potassium carbonate (2.4 g, 17.34 mmol) were added. The mixture was stirred in an oil bath at 55 °C for 18 h. The reaction proceeded completely as monitored by TLC. The mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to prepare slurry. The slurry was purified by column chromatography (petroleum ether) to give compound 4 (2.5 g, 87%).

[0048] Synthesis of compound I-5

[0049] Compound I-4 (1 g, 4.01 mmol), compound 3 (1.82 g, 4.81 mmol), 1,1′-bis(diphenylphosphine)ferrocene palladium dichloride (147 mg, 0.2 mmol), and potassium carbonate (1.11 g, 8.02 mmol) were dissolved in 1,4-dioxane (15 mL) and water (3 mL) under nitrogen balloon protection and stirred in an oil bath at 80 °C for 12 h. TLC analysis showed that the reaction proceeds were complete. The mixture was filtered through diatomaceous earth, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (petroleum ether:ethyl acetate = 20:1) to give compound 3 (1.3 g, 77%).

[0050] Synthesis of compound I-6

[0051] Compound I-5 (800 mg, 1.90 mmol) was dissolved in tetrahydrofuran (10 mL), and lithium aluminum hydride (216 mg, 5.69 mmol) was added in portions under ice bath conditions. The reaction was carried out at 0 °C for 1 h. The reaction was monitored by TLC until the starting material was completely reacted. The reaction was quenched dropwise with methanol, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to prepare slurry. The slurry was purified by column chromatography (petroleum ether:ethyl acetate = 2:1) to give compound 6 (670 mg, 90%).

[0052] Synthesis of compound I-7

[0053] Compound I-6 (600 mg, 1.52 mmol) was dissolved in 10 mL of dichloromethane. Sodium bicarbonate (256 mg, 3.05 mmol) and Desmartin reagent (969 mg, 2.29 mmol) were added under ice bath conditions. The mixture was then moved to room temperature and reacted for 1 h. The reaction was monitored by TLC until it was complete. The reaction was quenched by adding saturated sodium thiosulfate solution and saturated sodium bicarbonate solution. The mixture was extracted with dichloromethane, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to form sand. The sand was purified by column chromatography (petroleum ether:ethyl acetate = 4:1) to give compound 7 (580 mg, 97%).

[0054] Synthesis of compound I-8

[0055] Compound I-7 (100 mg, 0.255 mmol) was dissolved in 3 mL of N,N-dimethylformamide, and morpholine (44 mg, 0.51 mmol), potassium carbonate (71 mg, 0.51 mmol), and potassium iodide (8 mg, 0.051 mmol) were added sequentially. The mixture was stirred in an oil bath at 80 °C for 12 h. The reaction was monitored by TLC until it was complete. The mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to prepare slurry. The slurry was purified by column chromatography (petroleum ether:ethyl acetate = 1:1) to give compound 7 (86 mg, 76%).

[0056] Synthesis of Compound 1

[0057] Compound I-7 (60 mg) and ethanolamine (17 mg) were dissolved in methanol (2 mL) and dichloromethane (2 mL), 1 drop of glacial acetic acid was added, and the mixture was stirred at room temperature for 1 h. Then sodium cyanoborohydride (26 mg) was added, and the mixture was stirred at room temperature for another 12 h. The reaction was monitored by TLC until it was complete. The mixture was washed with saturated sodium bicarbonate solution, extracted with dichloromethane, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to prepare slurry. The slurry was then subjected to column chromatography (dichloromethane:methanol = 20:1) to obtain compound JC-1 (56 mg, 85%). 1 H NMR (400MHz, Chloroform-d) δ8.23 (s, 1H), 8.14 (d, J=7.0Hz, 1H), 7.82-7.72 (m, 2H), 7.59 (d, J=8.3Hz, 1H), 7.49(d, J=7.8Hz, 2H), 7.37(t, J=7.8Hz, 1H), 7.24-7.15(m, 2H), 6.96-6.88(m, 1H), 4.10 (t, J=6.3Hz, 2H), 3.92 (s, 2H), 3.72 (q, J=6.4, 5.7Hz, 6H), 2.85 (t, J=5.1Hz, 2H), 2. 55 (t, J=7.3Hz, 4H), 2.48 (t, J=4.7Hz, 2H), 2.01 (p, J==6.5Hz, 2H); MS (ESI, m / z): 488.6[M+H] + .

[0058] Example 2, Preparation of Compound 2

[0059]

[0060] The synthesis method is the same as in Example 1. 1H NMR (400MHz, Methanol-d4) δ8.26-8.19 (m, 1H), 8.17-8.08 (m, 1H), 7.89-7.82 (m, 1H), 7.74 (t, J=8.2H z, 1H), 7.68-7.50 (m, 3H), 7.41-7.30 (m, 1H), 7.27-7.14 (m, 2H), 6.97-6.87 (m, 1H), 4.38 (dp, J=8.7, 3. 1Hz, 1H), 4.09 (t, J=6.1Hz, 2H), 3.92 (d, J=5.5Hz, 2H), 3.71 (t, J=5.6Hz, 2H), 2.98-2.56 (m, 8H), 2.22 -2.09 (m, 1H), 2.09-1.97 (m, 2H), 1.76 (dddd, J==13.1, 8.0, 5.3, 2.9Hz, 1H); MS (ESI, m / z): 488.4[M+H] + .

[0061] Example 3, Preparation of Compound 3

[0062]

[0063] The synthesis method is the same as in Example 1. 1 H NMR (400MHz, Chloroform-d) δ8.23 (d, J=1.9Hz, 1H), 8.14 (dt, J=6.9, 1.9Hz, 1H), 7.81-7.71 (m, 2H), 7.58 (dd, J= 8.3, 1.7Hz, 1H), 7.54-7.46 (m, 2H), 7.36 (t, J=7.9Hz, 1H), 7.23-7.11 (m, 2H), 6.90 (dd, J=8.2, 2.5Hz, 1H), 4.08 (t , J=6.2Hz, 2H), 3.92 (s, 2H), 3.72 (q, J=5.2, 4.6Hz, 3H), 2.84 (dt, J=8.9, 4.8Hz, 4H), 2.72 (s, 3H), 2.58 (t, J=7.4 Hz, 2H), 2.23 (t, J=10.1Hz, 2H), 2.09-1.90 (m, 4H), 1.63 (dtd, J=13.0, 9.2, 3.6Hz, 2H); MS (ESI, m / z): 502.8[M+H] + .

[0064] Example 4, Preparation of Compound 4

[0065]

[0066] The synthesis method is the same as in Example 1.1 H NMR (400MHz, Chloroform-d) δ8.25 (d, J=1.7Hz, 1H), 8.16 (dt, J=7.2, 1.8Hz, 1H), 7.77 (d, J=8.2 Hz, 1H), 7.57-7.46 (m, 3H), 7.31 (dd, J=8.1, 1.6Hz, 1H), 7.22 (t, J=7.9Hz, 1H), 6.90 (t, J=8.4Hz , 2H), 4.09 (t, J=6.1Hz, 2H), 3.94 (s, 2H), 3.73 (dt, J=10.3, 4.9Hz, 6H), 2.91-2.83 (m, 2H), 2.64 -2.54(m, 4H), 2.49(t, J=4.6Hz, 4H), 2.16(s, 3H), 2.10-1.98(m, 2H); MS (ESI, m / z): 502.5[M+H] + .

[0067] Example 5: Preparation of Compound 5

[0068]

[0069] The synthesis method is the same as in Example 1. 1 H NMR (400MHz, Methanol-d4) δ8.30 (d, J=7.1Hz, 1H), 8.19 (t, J=6.5Hz, 1H), 7.74 (dd, J=8.0, 3.4Hz, 1H), 7.66 ( d, J=7.4Hz, 1H), 7.63-7.50 (m, 2H), 7.31 (td, J=8.7, 8.1, 3.0Hz, 1H), 7.20 (q, J=7.8, 7.1Hz, 1H), 6.94 (d, J=8 .0Hz, 1H), 6.86 (d, J=7.5Hz, 1H), 4.48 (tt, J=5.3, 2.4Hz, 1H), 4.20-4.00 (m, 4H), 3.77 (t, J=5.5Hz, 2H), 3.28 -2.88(m, 8H), 2.30-2.14(m, 3H), 2.13(d, J=2.8Hz, 3H), 1.91(p, J=6.5, 5.4Hz, 1H); MS (ESI, m / z): 502.2[M+H] + .

[0070] Example 6, Preparation of Compound 6

[0071]

[0072] The synthesis method is the same as in Example 1. 1H NMR (400MHz, Methanol-d4) δ8.31-8.24 (m, 1H), 8.17 (tt, J=7.5, 1.8Hz, 1H), 7.75 (dd, J=9.2, 4.2Hz, 1H), 7.69-7.50 (m, 3H), 7.33 (ddd, J=9 .7, 6.6, 1.7Hz, 1H), 7.20 (t, J=7.8Hz, 1H), 6.94 (d, J=8.2Hz, 1H), 6.87 (d, J=7.5Hz, 1H), 4.08 (q, J=6.1, 4.6Hz, 2H), 3.97 (d, J=6.6Hz, 2H), 3.74 (t, J=5.6Hz, 2H), 3.68 (dt, J=9.8, 5.4Hz, 1H), 2.92 (dt, J=10.0, 4.5Hz, 2H), 2.83 (t, J=5.5Hz, 2H), 2.66 (t, J=7.8Hz, 2H), 2.31 (d, J=1 1.0Hz, 2H), 2.14 (d, J=3.6Hz, 3H), 2.09-2.01 (m, 2H), 1.91 (dt, J=13.5, 4.1Hz, 2H), 1.63 (qd, J=9.6, 4.7Hz, 2H); MS (ESI, m / z): 516.1[M+H] + .

[0073] Example 7, Preparation of Compound 7

[0074]

[0075] The synthesis method is the same as in Example 1. 1H NMR (400MHz, Chloroform-d) δ8.25 (d, J=1.8Hz, 1H), 8.17 (dt, J=6.8, 1.9Hz, 1H), 7.78 (d, J=8.1Hz, 1H), 7.57-7.46 (m, 3H), 7.31 (dd, J=8.2, 1.5Hz, 1H), 7.22 (t, J=7.9Hz, 1H), 6.90 (ddd, J=12.5, 8.0, 1.1Hz, 2H), 4.08 (t, J=6.1Hz, 2H), 3.94 (s, 2H), 3.76-3.69 (m, 2H), 3.51 (d, J=6.4Hz, 2H), 3.04 (dt, J=11.7, 3.3Hz, 2H), 2.90-2.83 (m, 2H), 2.62 (dd, J=13.4, 6.2Hz, 5H), 2. 17(s, 3H), 2.12-1.97(m, 4H), 1.82-1.73(m, 2H), 1.58-1.48(m, 1H), 1.36(td, J=12.4, 3.8Hz, 2H); MS (ESI, m / z): 530.5[M+H] + .

[0076] Example 8, Preparation of Compound 8

[0077]

[0078] The synthesis method is the same as in Example 1. 1 H NMR (400MHz, Chloroform-d) δ8.24 (d, J=2.0Hz, 1H), 8.16 (dt, J=6.5, 2.0Hz, 1H), 7.77 (d, J=8.2Hz, 1H), 7.56-7.45 (m, 3H), 7.30 (dd, J=8.2, 1.6Hz, 1H), 7.20 (t, J=7.9Hz, 1H), 6.88 (dd, J=17.3, 7.9Hz, 2H), 4.46 (p, J=5.8Hz, 1H), 4.05 (t, J=6.1Hz, 2H), 3.93 (s, 2H), 3.79-3.60 (m, 4H), 3.05-2.93 (m, 2H), 2.90-2.82 ( m, 2H), 2.72 (t, J=7.3Hz, 2H), 2.51 (s, 3H), 2.15 (s, 3H), 1.91 (p, J=6.6Hz, 2H); MS (ESI, m / z): 488.7[M+H] + .

[0079] Example 9, Preparation of Compound 9

[0080]

[0081] The synthesis method is the same as in Example 1. 1 H NMR (400MHz, Chloroform-d) δ8.24 (d, J=1.8Hz, 1H), 8.16 (dt, J=6.8, 1.9Hz, 1H), 7.77 (d, J=8.1Hz, 1H), 7.55-7.47 (m, 3H), 7 .30 (dd, J=8.2, 1.5Hz, 1H), 7.21 (t, J=7.9Hz, 1H), 6.94-6.84 (m, 2H), 4.38 (ddt, J=7.4, 4.9, 2.2Hz, 1H), 4.09 (t, J=6.1Hz, 2H ), 3.93 (s, 2H), 3.76-3.67 (m, 2H), 3.01 (td, J=8.7, 5.1Hz, 1H), 2.90-2.79 (m, 3H), 2.75 (t, J=7.5Hz, 2H), 2.63-2.59 (m, 4H), 2.39 (td, J=9.0, 6.5Hz, 1H), 2.27-2.18 (m, 1H), 2.15 (s, 3H), 2.12-2.05 (m, 2H), 1.85-1.76 (m, 1H); MS (ESI, m / z): 502.5[M+H] + .

[0082] Example 10, Preparation of Compound 10

[0083]

[0084] The synthesis method is the same as in Example 1. 1H NMR (400MHz, Chloroform-d) δ8.25 (d, J=1.8Hz, 1H), 8.18 (dt, J=6.7, 1.7Hz, 1H), 7.75 (d, J=8.1Hz, 1H), 7.58- 7.46 (m, 3H), 7.28 (dd, J=8.0, 1.4Hz, 1H), 7.20 (t, J=7.8Hz, 1H), 6.96-6.83 (m, 2H), 4.28 (ddt, J=7.2, 4.5, 2.0 Hz, 1H), 4.06 (t, J=6.0Hz, 2H), 3.91 (s, 2H), 3.78-3.63 (m, 2H), 3.39 (s, 3 H), 3.00 (td, J=8.5, 5.0Hz, 1H), 2.91-2.77 (m, 3H), 2.73 (t, J=7.2Hz, 2H) , 2.62-2.55 (m, 3H), 2.40 (td, J=9.1, 6.4Hz, 1H), 2.25-2.15 (m, 1H), 2.15 (s, 3H), 2.11-2.03 (m, 2H), 1.83-1.73 (m, 1H); MS (ESI, m / z): 544.6[M+H] + .

[0085] Example 11, Preparation of Compound 11

[0086]

[0087] The synthesis method is the same as in Example 1. 1H NMR (400MHz, Chloroform-d) δ8.24 (d, J=1.8Hz, 1H), 8.17 (dt, J=6.7, 1.7Hz, 1H), 7.74 (d, J=8.1Hz, 1H), 7.57- 7.45 (m, 3H), 7.26 (dd, J=8.0, 1.4Hz, 1H), 7.21 (t, J=7.7Hz, 1H), 6.95-6.83 (m, 2H), 4.27 (ddt, J=7.2, 4.4, 2.0 Hz, 1H), 4.05 (t, J=6.1Hz, 2H), 3.90 (s, 2H), 3.77-3.63 (m, 2H), 3.39 (s, 3 H), 3.01 (td, J=8.4, 5.0Hz, 1H), 2.91-2.78 (m, 3H), 2.72 (t, J=7.2Hz, 2H) , 2.62-2.57 (m, 3H), 2.41 (td, J=9.0, 6.4Hz, 1H), 2.23-2.15 (m, 1H), 2.15 (s, 3H), 2.11-2.03 (m, 2H), 1.83-1.72 (m, 1H); MS (ESI, m / z): 544.5[M+H] + .

[0088] Example 12, Preparation of Compound 12

[0089]

[0090] The synthesis method is the same as in Example 1. 1H NMR (400MHz, Methanol-d4) δ8.30-8.24 (m, 1H), 8.16 (tt, J=7.5, 1.8Hz, 1H), 7.74 (dd, J=9.1, 4.2Hz, 1H), 7.68-7.50 (m, 3H), 7.33 (ddd, J=9.7 , 6.6, 1.7Hz, 1H), 7.21 (t, J = 7.7Hz, 1H), 6.93 (d, J = 8.2Hz, 1H), 6.88 (d, J = 7.3Hz, 1H), 4.08 (q, J = 6.1, 4.6Hz, 2H), 3.97 (d, J = 6.5Hz, 2H), 3.76 ( t, J=5.6Hz, 2H), 3.66 (dt, J=9.8, 5.3Hz, 1H), 3.37 (s, 3H), 2.93 (dt, J=9.8, 4.5Hz, 2H), 2.84 (t, J=5.5Hz, 2H), 2.66 (t, J=7.8Hz, 2H), 2.30 (d, J=11.0Hz, 2H), 2.15 (d, J=3.6Hz, 3H), 2.09-2.00 (m, 2H), 1.91 (dt, J=13.5, 4.1Hz, 2H), 1.63 (qd, J=9.6, 4.7Hz, 2H); MS (ESI, m / z): 558.2[M+H] + .

[0091] Example 13, Preparation of Compound 13

[0092]

[0093] The synthesis method is the same as in Example 1. 1H NMR (400MHz, Methanol-d4) δ8.24 (d, J=1.8Hz, 1H), 8.18 (dt, J=6.5, 1.7Hz, 1H), 7.76 (d, J=8.1Hz, 1H), 7.56-7.46 (m, 3H), 7. 26 (dd, J=8.0, 1.5Hz, 1H), 7.20 (t, J=7.8Hz, 1H), 6.94-6.81 (m, 2H), 4.26 (ddt, J=7.1, 4.3, 2.0Hz, 1H), 4.04 (t, J=6.0Hz, 2H) , 3.91 (s, 2H), 3.78-3.63 (m, 2H), 3.00 (td, J=8.5, 5.0Hz, 1H), 2.87-2.74 (m, 3H), 2.71 (t, J=7.2Hz, 2H), 2.60-2.53 (m, 2H), 2 .40 (td, J=9.1, 6.4Hz, 1H), 2.25-2.13 (m, 1H), 2.15 (s, 3H), 2.11-2.03 (m, 1H), 1.81-1.72 (m, 1H); MS (ESI, m / z): 530.1[M+H] + .

[0094] Example 14, Preparation of Compound 14

[0095]

[0096] The synthesis method is the same as in Example 1. 1 H NMR (400MHz, Methanol-d4) δ8.24 (d, J=1.8Hz, 1H), 8.18 (dt, J=6.5, 1.7Hz, 1H), 7.76 (d, J=8.1Hz, 1H), 7.55- 7.44 (m, 3H), 7.26 (dd, J=8.0, 1.5Hz, 1H), 7.20 (t, J=7.8Hz, 1H), 6.93-6.80 (m, 2H), 4.25 (ddt, J=7.0, 4.3, 2.0 Hz, 1H), 4.02 (t, J=6.0Hz, 2H), 3.93 (s, 2H), 3.76-3.63 (m, 2H), 3.02 (td, J=8.5, 5.0Hz, 1H), 2.87-2.72 (m, 3H), 2.71 (t, J=7.2Hz, 2H), 2.63 -2.53 (m, 2H), 2.40 (td, J=9.1, 6.4Hz, 1H), 2.25-2.12 (m, 1H), 2.15 (s, 3H), 2.10-2.03 (m, 1H), 1.81-1.71 (m, 1H); MS (ESI, m / z): 530.3[M+H] + .

[0097] Example 15, Preparation of Compound 15

[0098]

[0099] The synthesis method is the same as in Example 1. 1 H NMR (400MHz, Methanol-d4) δ8.30-8.24 (m, 1H), 8.15 (tt, J=7.5, 1.8Hz, 1H), 7.73 (dd, J=9.1, 4.1Hz, 1H), 7.65-7.50 (m, 3H), 7.32 (ddd, J= 9.7, 6.5, 1.7Hz, 1H), 7.20 (t, J=7.6Hz, 1H), 6.93 (d, J=8.2Hz, 1H), 6.88 (d, J=7.3Hz, 1H), 4.06 (q, J=6.0, 4.4Hz, 2H), 3.97 (d, J=6.5Hz, 2H) , 3.75 (t, J=5.6Hz, 2H), 3.64 (dt, J=9.6, 5.3Hz, 1H), 2.93 (dt, J=9.8, 4.5Hz, 2H), 2.84 (t, J=5.5Hz, 2H), 2.66 (t, J=7.8Hz, 2H), 2.30 (d, J=1 1.0Hz, 2H), 2.15 (d, J=3.6Hz, 3H), 2.14-2.03 (m, 2H), 1.91 (dt, J=13.5, 4.1Hz, 2H), 1.64 (qd, J=9.6, 4.6Hz, 2H); MS (ESI, m / z): 544.6[M+H] + .

[0100] Example 16, Preparation of Compound 16

[0101]

[0102] The synthesis method is the same as in Example 1. 1H NMR (400MHz, Chloroform-d) δ8.25 (d, J=1.7Hz, 1H), 8.15 (dt, J=7.1, 1.6Hz, 1H), 7.76 (d, J=8.1 Hz, 1H), 7.55-7.44 (m, 3H), 7.30 (dd, J=8.0, 1.4Hz, 1H), 7.22 (t, J=7.9Hz, 1H), 6.91 (t, J=8.3Hz , 2H), 4.09 (t, J=6.1Hz, 2H), 3.93 (s, 2H), 3.72 (dt, J=10.2, 4.6Hz, 6H), 2.90-2.80 (m, 3H), 2.65 -2.51(m, 4H), 2.51(t, J=4.6Hz, 4H), 2.15(s, 3H), 2.10-1.98(m, 2H); MS (ESI, m / z): 501.6[M+H] + .

[0103] Example 17, Preparation of Compound 17

[0104]

[0105] The synthesis method is the same as in Example 1. 1 H NMR (400MHz, Methanol-d4) δ8.31-8.25 (m, 1H), 8.16 (tt, J=7.3, 1.8Hz, 1H), 7.71 (dd, J=9.3, 4.1Hz, 1H), 7.63-7.47 (m, 3H), 7.30 (ddd, J= 9.4, 6.2, 1.6Hz, 1H), 7.16 (t, J=7.2Hz, 1H), 6.90 (d, J=8.2Hz, 1H), 6.87 (d, J=7.3Hz, 1H), 4.04 (q, J=6.3, 4.1Hz, 2H), 3.94 (d, J=6.3Hz, 2H) , 3.72 (t, J=5.5Hz, 2H), 3.63 (dt, J=9.5, 5.3Hz, 1H), 2.92 (dt, J=9.6, 4.4Hz, 2H), 2.83 (t, J=5.6Hz, 2H), 2.64 (t, J=7.8Hz, 2H), 2.30 (d, J=1 1.0Hz, 2H), 2.15 (d, J=3.6Hz, 3H), 2.27-2.18 (m, 2H), 1.91 (dt, J=13.5, 4.1Hz, 2H), 1.64 (qd, J=9.6, 4.6Hz, 2H); MS (ESI, m / z): 515.7[M+H] + .

[0106] Example 18, Preparation of Compound 18

[0107]

[0108] The synthesis method is the same as in Example 1. 1 H NMR (400MHz, Methanol-d4) δ8.29 (d, J=7.0Hz, 1H), 8.18 (t, J=6.5Hz, 1H), 7.76 (dd, J=8.0, 3.3Hz, 1H), 7.68 ( d, J=7.4Hz, 1H), 7.65-7.51 (m, 2H), 7.31 (td, J=8.7, 8.1, 3.0Hz, 1H), 7.22 (q, J=7.9, 7.3Hz, 1H), 6.95 (d, J=8 .1Hz, 1H), 6.86 (d, J=7.5Hz, 1H), 4.50 (tt, J=5.4, 2.6Hz, 1H), 4.22-4.05 (m, 4H), 3.76 (t, J=5.4Hz, 2H), 3.32 -2.92 (m, 8H), 2.31-2.13 (m, 3H), 2.14 (d, J=2.8Hz, 3H), 1.95 (p, J=6.4, 5.4Hz, 1H); MS (ESI, m / z): 501.6[M+H] + .

[0109] Example 19, Preparation of Compound 19

[0110]

[0111] The synthesis method is the same as in Example 1. 1 H NMR (400MHz, Chloroform-d) δ8.26 (d, J=2.0Hz, 1H), 8.18 (dt, J=7.1, 1.8Hz, 1H), 7.61 (d, J=8.1Hz, 1H), 7.5 6-7.47 (m, 2H), 7.21 (t, J=7.9Hz, 1H), 7.15 (d, J=8.1Hz, 1H), 6.90 (d, J=8.1Hz, 1H), 6.80 (d, J=7.6Hz, 1H), 4 .10(q, J=5.9Hz, 2H), 3.95(s, 2H), 3.74(dt, J=8.1, 4.8Hz, 6H), 2.88(t, J=5.1Hz, 2H), 2.75(s, 2H), 2.60(t, J=7.4Hz, 2H), 2.51(d, J=4.7Hz, 4H), 2.32(s, 3H), 2.10-2.01(m, 2H), 1.97(s, 3H); MS (ESI, m / z): 516.4[M+H] + .

[0112] Example 20, Preparation of Compound 20

[0113]

[0114] The synthesis method is the same as in Example 1. 1 H NMR (400MHz, Methanol-d4) δ8.15 (dd, J=6.7, 2.5Hz, 1H), 8.09-8.00 (m, 1H), 7.55-7.39 (m, 3H), 7.13-7.03 (m, 1H ), 7.03-6.95 (m, 1H), 6.84 (d, J=7.8Hz, 1H), 6.62 (t, J=6.7Hz, 1H), 4.28 (ddq, J=8.4, 5.3, 2.7Hz, 1H), 3.99 (t, J= 5.9Hz, 2H), 3.83 (d, J=7.0Hz, 2H), 3.61 (t, J=5.6Hz, 2H), 2.83 (dt, J=10.5, 5.3Hz, 1H), 2.79-2.63 (m, 5H), 2.62- 2.50 (m, 2H), 2.21-2.10 (m, 3H), 2.10-1.91 (m, 3H), 1.81 (d, J=3.4Hz, 3H), 1.67 (m, 1H); MS (ESI, m / z): 516.6[M+H] + .

[0115] Example 21, Preparation of Compound 21

[0116]

[0117] The synthesis method is the same as in Example 1. 1 H NMR (300MHz, Methanol-d4) δ8.29 (d, J=1.7Hz, 1H), 8.20 (dt, J=7.3, 1.7Hz, 1H), 7.69-7.54 (m, 3H), 7 .28-7.09 (m, 2H), 6.97 (dd, J=8.3, 1.1Hz, 1H), 6.75 (dd, J=7.6, 1.0Hz, 1H), 4.11 (t, J=6.0Hz, 2H), 3. 99(s, 2H), 3.80-3.62(m, 3H), 2.89(dt, J=26.5, 6.0Hz, 4H), 2.74-2.63(m, 2H), 2.29(s, 5H), 2.09(dp , J=12.1, 6.8, 6.2Hz, 2H), 1.94 (s, 5H), 1.63 (dtd, J=13.2, 9.5, 3.7Hz, 2H); MS (ESI, m / z): 530.1[M+H] + .

[0118] Example 22, Preparation of Compound 22

[0119]

[0120] The synthesis method is the same as in Example 1. 1 H NMR (300MHz, Methanol-d4) δ8.28 (d, J=1.6Hz, 1H), 8.21 (dt, J=7.1, 1.6Hz, 1H), 7.68-7.52 (m, 3H), 7 .29-7.07 (m, 2H), 6.98 (dd, J=8.2, 1.0Hz, 1H), 6.76 (dd, J=7.5, 1.2Hz, 1H), 4.09 (t, J=6.2Hz, 2H), 3. 98 (s, 2H), 3.80-3.63 (m, 3H), 3.50 (d, J=6.2, 2H), 2.89 (dt, J=26.5, 6.0Hz, 4H), 2.74-2.63 (m, 2H), 2 .29 (s, 5H), 2.09 (m, 2H), 1.94 (s, 5H), 1.61 (dtd, J=13.2, 9.5, 3.7Hz, 2H); MS (ESI, m / z): 544.6[M+H] + .

[0121] Example 23, Preparation of Compound 23

[0122]

[0123] The synthesis method is the same as in Example 1. 1 H NMR (400MHz, Chloroform-d) δ8.27 (d, J=1.8Hz, 1H), 8.22 (dt, J=7.3, 1.7Hz, 1H), 7.67-7.54 (m, 3 H), 7.27-7.10 (m, 2H), 6.96 (dd, J=8.1, 1.3Hz, 1H), 6.74 (dd, J=7.1, 1.0Hz, 1H), 4.43 (p, J=5.8Hz, 1H), 4.02(t, J=6.1Hz, 2H), 3.91(s, 2H), 3.75-3.62(m, 4H), 3.03-2.91(m, 2H), 2.92-2.80(m, 2H), 2.76 (t, J=7.1Hz, 2H), 2.34 (s, 3H), 1.96 (s, 3H), 1.96 (p, J=6.3Hz, 2H); MS (ESI, m / z): 502.5[M+H] + .

[0124] Example 24, Preparation of Compound 24

[0125]

[0126] The synthesis method is the same as in Example 1. 1 H NMR (400MHz, Chloroform-d) δ8.30 (d, J=2.0Hz, 1H), 8.20 (dt, J=7.0, 1.4Hz, 1H), 7.71-7.60 (m, 3H), 7.2 3-7.05 (m, 2H), 6.94 (dd, J=7.7, 1.1Hz, 1H), 6.73 (dd, J=7.5, 1.3Hz, 1H), 4.37 (p, J=5.6Hz, 1H), 4.05 (t, J=6.0Hz, 2H), 3.88 (s, 2H), 3.65-3.59 (m, 2H), 3.01 (td, J=8.0, 4.7Hz, 1H), 2.84-2.73 (m, 3H), 2.71 (t, J =7.2Hz, 2H), 2.61-2.50(m, 4H), 2.30(s, 5H), 2.01(s, 5H), 1.85-1.76(m, 1H); MS (ESI, m / z): 516.7[M+H] + .

[0127] Example 25, Preparation of Compound 25

[0128]

[0129] The synthesis method is the same as in Example 1. 1 H NMR (400MHz, Chloroform-d) δ8.25 (d, J=1.7Hz, 1H), 8.18 (dt, J=6.8, 1.3Hz, 1H), 7.64-7.43 (m, 3H), 7.27- 7.10 (m, 2H), 6.96 (dd, J=8.1, 1.3Hz, 1H), 6.78 (dd, J=7.1, 1.0Hz, 1H), 4.36 (p, J=5.3Hz, 1H), 4.14 (t, J=6.0 Hz, 2H), 3.95 (s, 2H), 3.81-3.72 (m, 2H), 3.35 (s, 3H), 3.01 (td, J=8.5, 5.0Hz, 1H), 2.91-2.77 (m, 3H), 2.73 ( t, J=7.2Hz, 2H), 2.65-2.48(m, 4H), 2.33(s, 5H), 2.05(s, 5H), 1.83-1.73(m, 1H); MS (ESI, m / z): 558.8[M+H] + .

[0130] Example 26, Preparation of Compound 26

[0131]

[0132] The synthesis method is the same as in Example 1. 1 H NMR (400MHz, Chloroform-d) δ8.24 (d, J=1.7Hz, 1H), 8.16 (dt, J=6.6, 1.1Hz, 1H), 7.60-7.41 (m, 3H), 7.24- 7.08 (m, 2H), 6.95 (dd, J=7.8, 1.0Hz, 1H), 6.76 (dd, J=7.0, 1.3Hz, 1H), 4.34 (p, J=5.5Hz, 1H), 4.13 (t, J=6.0 Hz, 2H), 3.95 (s, 2H), 3.81-3.71 (m, 2H), 3.35 (s, 3H), 3.00 (td, J=8.4, 5.0Hz, 1H), 2.91-2.75 (m, 3H), 2.72 ( t, J=7.2Hz, 2H), 2.66-2.48 (m, 4H), 2.33 (s, 5H), 2.05 (s, 5H), 1.86-1.74 (m, 1H); MS (ESI, m / z): 558.3[M+H] + .

[0133] Example 27, Preparation of Compound 27

[0134]

[0135] The synthesis method is the same as in Example 1. 1 H NMR (400MHz, Methanol-d4) δ8.29-8.23 (m, 1H), 8.15 (tt, J=7.1, 1.5Hz, 1H), 7.64-7.42 (m, 3H), 7.28-7.13 (m, 2 H), 6.91 (d, J=8.1Hz, 1H), 6.83 (d, J=7.3Hz, 1H), 4.28 (q, J=6.1, 4.6Hz, 2H), 3.97 (d, J=6.5Hz, 2H), 3.71 (t, J=5 .6Hz, 2H), 3.62 (dt, J=9.8, 5.3Hz, 1H), 3.32 (s, 3H), 2.91 (dt, J=9.5, 4.1Hz, 2H), 2.80 (t, J=5.5Hz, 2H), 2.61-2 .43 (m, 4H), 2.31 (s, 5H), 2.04 (s, 5H), 1.82-1.71 (m, 1H) 1.63 (qd, J=9.3, 4.1Hz, 1H); MS (ESI, m / z): 558.2[M+H] + .

[0136] Example 28, Preparation of Compound 28

[0137]

[0138] The synthesis method is the same as in Example 1. 1 H NMR (400MHz, Methanol-d4) δ8.25 (d, J=1.9Hz, 1H), 8.15 (dt, J=6.3, 1.5Hz, 1H), 7.51-7.44 (m, 3H), 7.25 (dd, J =8.1, 1.4Hz, 1H), 7.23 (t, J = 7.5Hz, 1H), 6.90-6.80 (m, 2H), 4.24 (m, 1H), 4.01 (t, J = 5.0Hz, 2H), 3.88 (s, 2H), 3 .75-3.60 (m, 2H), 3.30 (s, 3H), 2.98 (td, J=8.3, 4.0Hz, 1H), 2.88-2.72 (m, 2H), 2.67 (t, J=6.2Hz, 2H), 2.64-2. 42 (m, 2H), 2.34 (s, 3H), 2.07 (s, 3H), 1.81-1.75 (m, 1H) 1.63 (qd, J=8.7, 4.0Hz, 1H); MS (ESI, m / z): 544.1[M+H] + .

[0139] Example 29, Preparation of Compound 29

[0140]

[0141] The synthesis method is the same as in Example 1. 1 H NMR (400MHz, Methanol-d4) δ8.25 (d, J=1.9Hz, 1H), 8.15 (dt, J=6.3, 1.5Hz, 1H), 7.52-7.45 (m, 3H), 7.26 (dd, J =8.0, 1.4Hz, 1H), 7.22 (t, J = 7.5Hz, 1H), 6.92-6.78 (m, 2H), 4.26 (m, 1H), 4.00 (t, J = 4.8Hz, 2H), 3.86 (s, 2H), 3 .75-3.61 (m, 2H), 3.30 (s, 3H), 2.96 (td, J=8.0, 4.1Hz, 1H), 2.85-2.72 (m, 2H), 2.65 (t, J=6.0Hz, 2H), 2.60-2. 45 (m, 2H), 2.34 (s, 3H), 2.07 (s, 3H), 1.80-1.75 (m, 1H) 1.64 (qd, J=8.5, 4.1Hz, 1H); MS (ESI, m / z): 544.2[M[+H] + .

[0142] Example 30, Preparation of Compound 30

[0143]

[0144] The synthesis method is the same as in Example 1. 1 H NMR (400MHz, Methanol-d4) δ8.32-8.26 (m, 1H), 8.16 (tt, J=7.0, 1.6Hz, 1H), 7.62-7.46 (m, 3H), 7.32 (m, 1H), 7.21 (t, J=7.7Hz, 1H), 6.93-6.74 (m, 2 H), 4.10 (q, J=5.6, 4.0Hz, 2H), 3.95 (d, J=6.0Hz, 2H), 3.70 (t, J=5.6Hz, 2H ), 3.60 (dt, J=9.0, 5.1Hz, 1H), 2.92 (dt, J=9.3, 4.8Hz, 2H), 2.82 (t, J=5.0 Hz, 2H), 2.62 (t, J=7.2Hz, 2H), 2.35 (s, 5H), 2.09 (s, 5H), 1.91 (dt, J=12.5, 4.1Hz, 2H), 1.64 (qd, J=9.2, 4.6Hz, 2H); MS (ESI, m / z): 558.6[M+H] + .

[0145] Example 31, Preparation of Compound 31

[0146]

[0147] The synthesis method is the same as in Example 1. 1 H NMR (400MHz, Chloroform-d) δ8.26 (d, J=1.6Hz, 1H), 8.15 (dt, J=7.0, 1.6Hz, 1H), 7.58-7 .41 (m, 3H), 7.28 (dd, J=8.2, 1.3Hz, 1H), 7.21 (t, J=7.9Hz, 1H), 6.91 (t, J=8.1Hz, 2H), 4. 05 (t, J=6.0Hz, 2H), 3.95 (s, 2H), 3.76 (dt, J=10.0, 4.8Hz, 6H), 2.95-2.75 (m, 3H), 2.62- 2.51 (m, 4H), 2.51 (t, J=4.6Hz, 4H), 2.32 (s, 3H), 2.12 (s, 5H); MS (ESI, m / z): 515.2[M+H] + .

[0148] Example 32, Preparation of Compound 32

[0149]

[0150] The synthesis method is the same as in Example 1. 1 H NMR (400MHz, Methanol-d4) δ8.29-8.24 (m, 1H), 8.14 (tt, J=7.0, 1.5Hz, 1H), 7.63-7.47 (m, 3H), 7.30 (ddd, J=9.4, 6.2, 1.6Hz, 1H), 7.16 (t, J=7.2Hz, 1H), 6.90 (d, J=8.2Hz, 1H), 6.87 (d, J=7.3Hz, 1H), 4.04 (q, J=6.3, 4.1Hz, 2H), 3.92 (d, J= 6.0Hz, 2H), 3.75 (t, J=5.2Hz, 2H), 3.62 (dt, J=9.5, 5.3Hz, 1H), 2.96 (dt, J=9.6, 4.1Hz, 2H), 2.81 (t, J=5.0Hz, 2H), 2.6 4(t, J=7.5Hz, 2H), 2.36(s, 5H), 2.27-2.18(m, 2H), 2.14(s, 5H), 1.64(qd, J=9.6, 4.6Hz, 2H); MS (ESI, m / z): 515.7[M+H] + .

[0151] Example 33, Preparation of Compound 33

[0152]

[0153] The synthesis method is the same as in Example 1. 1 H NMR (400MHz, Methanol-d4) δ8.26 (d, J=7.4Hz, 1H), 8.15 (t, J=6.2Hz, 1H), 7.67 (d, J=7.4Hz, 1H), 7 .62-7.51 (m, 2H), 7.32 (td, J=8.5, 8.0, 3.2Hz, 1H), 7.20 (q, J=7.3, 6.3Hz, 1H), 6.93 (d, J=8.1Hz, 1 H), 6.82 (d, J=8.0Hz, 1H), 4.48 (tt, J=5.0, 2.6Hz, 1H), 4.25-4.08 (m, 4H), 3.72 (t, J=5.4Hz, 2H), 3 .38-3.01 (m, 8H), 2.31 (m, 5H), 2.14 (s, 3H), 1.96 (p, J=6.0, 5.4Hz, 1H); MS (ESI, m / z): 515.1[M+H] + .

[0154] Example 34, Preparation of Compound 34

[0155]

[0156] The synthesis method is the same as in Example 1. 1 H NMR (400MHz, Methanol-d4) δ8.28 (d, J=7.0Hz, 1H), 8.14 (t, J=6.1Hz, 1H), 7.70 (dd, J=8.0, 3.5Hz, 1H), 7.62 (d, J=7.7Hz, 1H), 7.60-7.48 (m, 2H), 7.30 (td, J=8.5, 8.0, 3.3Hz, 1H), 7.16 (q, J=7.4, 7.0Hz, 1H), 6 .90 (d, J=8.3Hz, 1H), 6.83 (d, J=7.2Hz, 1H), 4.45 (tt, J=5.1, 2.4Hz, 1H), 4.15-4.02 (m, 4H), 3.75 (t, J= 5.5Hz, 2H), 3.27-2.90 (m, 8H), 2.32-2.16 (m, 3H), 1.93 (p, J=6.0, 5.1Hz, 1H); MS (ESI, m / z): 566.3[M+H] + .

[0157] Example 35, Preparation of Compound 35

[0158]

[0159] The synthesis method is the same as in Example 1. 1 H NMR (400MHz, Methanol-d4) δ8.27 (d, J=7.1Hz, 1H), 8.14 (t, J=6.0Hz, 1H), 7.71 (dd, J=8.0, 3.6 Hz, 1H), 7.63 (d, J=7.8Hz, 1H), 7.61-7.48 (m, 2H), 7.31 (td, J=8.4, 8.0, 3.3Hz, 1H), 7.14 (q, J= 7.4, 7.0Hz, 1H), 6.94 (d, J=8.3Hz, 1H), 6.82 (d, J=7.2Hz, 1H), 4.41 (tt, J=5.1, 2.3Hz, 1H), 4.1 5-4.03 (m, 4H), 3.76 (t, J=5.6Hz, 2H), 3.26-2.89 (m, 8H), 2.31-2.12 (m, 3H), 1.92 (p, J=6.0, 5.0 Hz, 1H); MS (ESI, m / z): 566.6[M+H] + .

[0160] Example 36, Preparation of Compound 36

[0161]

[0162] The synthesis method is the same as in Example 1. 1 H NMR (400MHz, Methanol-d4) δ8.24 (d, J=7.1Hz, 1H), 8.11 (t, J=6.3Hz, 1H), 7.74 (dd, J=7.9, 3.4Hz, 1H), 7.65 (d, J=7.3Hz, 1H), 7.62-7.48 (m, 2H), 7.33 (td, J=8.1, 7.3, 3.5Hz, 1H), 7.16 (q, J=7.9, 6.7Hz, 1H), 6 .92 (d, J=8.0Hz, 1H), 6.82 (d, J=7.2Hz, 1H), 4.40 (tt, J=5.1, 2.3Hz, 1H), 4.15-4.04 (m, 4H), 3.76 (t, J= 5.5Hz, 2H), 3.23-2.86 (m, 8H), 2.30-2.16 (m, 3H), 1.90 (p, J=5.6, 4.8Hz, 1H); MS (ESI, m / z): 566.1[M+H] + .

[0163] Example 37, Preparation of Compound 37

[0164]

[0165] The synthesis method is the same as in Example 1. 1H NMR (400MHz, Methanol-d4) δ8.28 (m, 1H), 8.17 (tt, J=7.5, 1.8Hz, 1H), 7.75 (dd, J=9.2, 4.2Hz, 1H), 7.69-7.50 (m, 3H), 7.33 (ddd, J=9.7, 6.6, 1.7Hz, 1H), 7.20 (t, J=7.8Hz, 1H), 6.94 (d, J=8.2Hz, 1H), 6.88 (d, J=7.5Hz, 1H), 4.09 (q, J=6.1, 4.6Hz, 2H), 3.95 (d, J=6.5Hz, 2H), 3. 73 (t, J=5.6Hz, 2H), 3.66 (dt, J=9.3, 5.1Hz, 1H), 2.93 (dt, J=10.1, 4.5Hz, 2H), 2.82 (t, J=5.5Hz, 2H), 2.68 (t, J=7.8Hz, 2H), 2.30 (d, J=11 .2Hz, 2H), 2.16 (d, J=3.6Hz, 3H), 2.11-2.01 (m, 2H), 1.89 (dt, J=13.5, 4.1Hz, 2H), 1.65 (qd, J=9.6, 4.6Hz, 2H); MS (ESI, m / z): 580.7[M+H] + .

[0166] Example 38, Preparation of Compound 38

[0167]

[0168] The synthesis method is the same as in Example 1. 1H NMR (400MHz, Methanol-d4) δ8.27 (m, 1H), 8.16 (tt, J=7.3, 1.7Hz, 1H), 7.74 (dd, J=9.2, 4.1Hz, 1H), 7.65-7.51 (m, 3H), 7.30 (ddd, J=9.4, 6.5, 1.7Hz, 1H), 7.21 (t, J=7.7Hz, 1H), 6.91 (d, J=8.1Hz, 1H), 6.83 (d, J=7.5Hz, 1H), 4.04 (q, J=6.0, 4.1Hz, 2H), 3.93 (d, J=6.3Hz, 2H), 3. 72 (t, J=5.5Hz, 2H), 3.67 (dt, J=9.3, 5.1Hz, 1H), 2.92 (dt, J=10.0, 4.5Hz, 2H), 2.86 (t, J=5.5Hz, 2H), 2.69 (t, J=7.8Hz, 2H), 2.32 (d, J=11 .0Hz, 2H), 2.16 (d, J=3.5Hz, 3H), 2.11-2.00 (m, 2H), 1.90 (dt, J=13.4, 4.1Hz, 2H), 1.65 (qd, J=9.6, 4.5Hz, 2H); MS (ESI, m / z): 536.5[M+H] + .

[0169] Example 39, Preparation of Compound 39

[0170]

[0171] The synthesis method is the same as in Example 1. 1H NMR (400MHz, Methanol-d4) δ8.27 (m, 1H), 8.16 (tt, J=7.2, 1.7Hz, 1H), 7.75 (dd, J=9.2, 4.3Hz, 1H), 7.65-7.52 (m, 3H), 7.30 (ddd, J=9.0, 6.0, 1.7Hz, 1H), 7.24 (t, J=7.2Hz, 1H), 6.91 (d, J=8.1Hz, 1H), 6.83 (d, J=7.5Hz, 1H), 4.04 (q, J=6.0, 4.1Hz, 2H), 3.92 (d, J=6.4Hz, 2H), 3. 75 (t, J=5.0Hz, 2H), 3.64 (dt, J=9.0, 5.1Hz, 1H), 2.92 (dt, J=10.2, 4.3Hz, 2H), 2.82 (t, J=5.5Hz, 2H), 2.65 (t, J=7.8Hz, 2H), 2.27 (d, J=10 .0Hz, 2H), 2.16 (d, J=3.2Hz, 3H), 2.11-2.05 (m, 2H), 1.95 (dt, J=13.0, 4.1Hz, 2H), 1.69 (qd, J=9.6, 4.0Hz, 2H); MS (ESI, m / z): 520.4[M+H] + .

[0172] Example 40, Preparation of Compound 40

[0173]

[0174] The synthesis method is the same as in Example 1. 1 H NMR (400MHz, Methanol-d4) δ8.26 (d, J=7.1Hz, 1H), 8.15 (t, J=6.5Hz, 1H), 7.71 (dd, J=8.0, 3.4Hz, 1H), 7.63 ( d, J=7.4Hz, 1H), 7.60-7.48 (m, 2H), 7.25 (td, J=8.4, 7.7, 3.0Hz, 1H), 7.17 (q, J=7.8, 7.0Hz, 1H), 6.93 (d, J=8 .0Hz, 1H), 6.86 (d, J=7.5Hz, 1H), 4.48 (tt, J=5.3, 2.4Hz, 1H), 4.15-4.00 (m, 2H), 3.73 (t, J=5.0Hz, 2H), 3.27 -2.84 (m, 8H), 2.35-2.18 (m, 3H), 2.11 (d, J=2.8Hz, 3H), 1.90 (p, J=6.0, 5.4Hz, 1H); MS (ESI, m / z): 488.4[M+H] + .

[0175] Example 41, Preparation of Compound 41

[0176]

[0177] The synthesis method is the same as in Example 1. 1 H NMR (400MHz, Methanol-d4) δ8.25 (d, J=7.5Hz, 1H), 8.12 (t, J=6.1Hz, 1H), 7.75 (dd, J=7.9, 3.2Hz, 1H), 7. 60 (d, J=7.0Hz, 1H), 7.58-7.41 (m, 2H), 7.23 (td, J=8.4, 7.2, 3.2Hz, 1H), 7.10 (q, J=7.5, 7.0Hz, 1H), 6.90 (d, J=8.2Hz, 1H), 6.85 (d, J=7.2Hz, 1H), 4.43 (tt, J=5.7, 2.0Hz, 1H), 4.25-4.05 (m, 6H), 3.70 (t, J=5.1Hz , 2H), 3.27-2.81 (m, 8H), 2.39-2.14 (m, 3H), 2.10 (d, J=2.8Hz, 3H), 1.88 (m, 1H); MS (ESI, m / z): 516.2[M+H] + .

[0178] Example 42, Preparation of Compound 42

[0179]

[0180] The synthesis method is the same as in Example 1. 1 H NMR (400MHz, Methanol-d4) δ8.23 (d, J=7.5Hz, 1H), 8.10 (t, J=5.5Hz, 1H), 7.71 (dd, J=7.8, 3.0Hz, 1H), 7 .60 (d, J=6.5Hz, 1H), 7.55-7.40 (m, 2H), 7.21 (td, J=8.0, 7.1, 3.2Hz, 1H), 7.08 (q, J=7.9, 7.0Hz, 1H), 6.9 2 (d, J=8.2Hz, 1H), 6.80 (d, J=7.2Hz, 1H), 4.47 (tt, J=5.2, 2.4Hz, 1H), 4.25-4.05 (m, 6H), 3.74-3.58 (m, 4H), 3.30-2.94(m, 8H), 2.34-2.14(m, 3H), 2.12(d, J=2.8Hz, 3H), 1.85(m, 1H); MS (ESI, m / z): 530.3[M+H] + .

[0181] Example 43, Preparation of Compound 43

[0182]

[0183] The synthesis method is the same as in Example 1. 1 H NMR (400MHz, Methanol-d4) δ8.27 (m, 1H), 8.15 (tt, J=7.3, 1.6Hz, 1H), 7.73 (dd, J=8.8, 4.1Hz, 1H), 7.69-7.52 (m, 3H), 7.35 (ddd, J= 9.3, 6.2, 1.4Hz, 1H), 7.15 (t, J=7.8Hz, 1H), 6.90 (d, J=8.2Hz, 1H), 6.82 (d, J=7.5Hz, 1H), 4.11 (q, J=6.0, 4.3Hz, 2H), 3.95 (d, J=6.6 Hz, 2H), 3.71 (t, J=5.1Hz, 2H), 3.65 (dt, J=9.3, 5.0Hz, 1H), 2.92 (dt, J=10.1, 4.5Hz, 2H), 2.63 (t, J=7.8Hz, 2H), 2.33 (d, J=11.0Hz, 2H), 2.14 (d, J=3.2Hz, 3H), 2.09-2.01 (m, 2H), 1.92 (dt, J=13.5, 4.1Hz, 2H), 1.65 (qd, J=9.2, 4.7Hz, 2H); MS (ESI, m / z): 502.2[M+H] + .

[0184] Example 44, Preparation of Compound 44

[0185]

[0186] The synthesis method is the same as in Example 1. 1H NMR (400MHz, Methanol-d4) δ8.26 (m, 1H), 8.15 (tt, J=7.2, 1.6Hz, 1H), 7.72 (dd, J=8.6, 4.7Hz, 1H), 7.66-7.50 (m, 3H), 7.36 (ddd, J=9.3, 6.1, 1.4Hz, 1H), 7.18 (t, J=7.8Hz, 1H), 6.92 (d, J=8.2Hz, 1H), 6.82 (d, J=7.5Hz, 1H), 4.12 (q, J=6.0, 4.5Hz, 2H), 3.92 (d, J=6.8Hz, 2H), 3.65 (m, 4H), 2.82-2.63 (m, 4H), 2.35 (m, 4H), 2.14 (d, J=3.1H z, 3H), 2.12-2.02 (m, 3H), 1.94 (dt, J=13.0, 4.2Hz, 2H), 1.67 (qd, J=9.2, 4.7Hz, 2H); MS (ESI, m / z): 530.5[M+H] + .

[0187] Example 45, Preparation of Compound 45

[0188]

[0189] The synthesis method is the same as in Example 1. 1 H NMR (400MHz, Methanol-d4) δ8.27 (m, 1H), 8.12 (tt, J=7.0, 1.6Hz, 1H), 7.75 (dd, J=8.6, 4.7Hz, 1H), 7.65-7.51 (m, 3H), 7.37 (ddd, J=9.0, 6.2, 1.4Hz, 1H), 7.15 (t, J=7.8Hz, 1H), 6.95 (d, J=8.2Hz, 1H), 6.80 (d, J=7.5Hz, 1H), 4.13 (q, J=6.5, 4.0Hz, 2H), 3.95 (d, J=6.8Hz, 2H), 3.66 (m, 4H), 2.83-2.62 (m, 4H), 2.35 (m, 4H), 2.15 (d, J=3.0H z, 3H), 2.17-2.00 (m, 5H), 1.95 (dt, J=13.0, 4.2Hz, 2H), 1.65 (qd, J=9.2, 4.7Hz, 2H); MS (ESI, m / z): 544.3[M+H] + .

[0190] Example 46

[0191] Tablet preparation

[0192] The compound 1 (50g) obtained in Example 1, hydroxypropyl methylcellulose E (150g), starch (200g), appropriate amount of povidone K30 and magnesium stearate (1g) were mixed, granulated and compressed into tablets.

[0193] In addition, according to the conventional formulation method of the 2020 edition of the Pharmacopoeia, the compounds obtained in Examples 1-45 can be given different pharmaceutical excipients to make capsules, powders, granules, pills, injections, syrups, oral preparations, inhalers, ointments, suppositories or patches, etc.

[0194] Application Examples

[0195] Pharmacological tests have demonstrated that the VISTA and PD-1 / PD-L1 inhibitory activities of the present invention can be used to prepare antitumor drugs. The following are the pharmacological test results of the compounds of the present invention:

[0196] I. Experiment on the binding ability of compounds to VISTA protein

[0197] (I) Experimental Equipment and Reagents

[0198] 1. The model used in this experiment is Biacore S200.

[0199] 2. S-series CM5 chips. Item numbers: 29-1049-88 (one chip), BR-1005-30 (three chips), 29-1496-03 (ten chips), purchased from GE Healthcare.

[0200] 3. Amino-coupled reagent kit. Catalog number: BR-1000-50, purchased from GE Healthcare.

[0201] 4. Buffer solution: 10x PBS-P+ (catalog number: 28-9950-84), purchased from GE Healthcare.

[0202] 5. Analytical grade DMSO, deionized water (filtered through a 0.22μm membrane).

[0203] 6. Protein: VISTA protein modified with glycosylation.

[0204] 7. Other consumables: 1.5ml EP tubes without caps (item number: BR-1002-87), rubber bottle cap type 2 (item number: BR-1004-11), 96-well plate (item number: BR-1005-03), 96-well plate sealing film (item number: 28-9758-16), purchased from GE Healthcare.

[0205] (II) Experimental Procedure

[0206] The binding affinity of the compound to the VISTA protein was tested using a Biacore S200 system and CM5 chip. A 10 mM stock solution of the compound was diluted with 1.05*PBS-P to 10 concentration gradients (5 μM, 2.5 μM, 1.25 μM, 0.625 μM, 0.3125 μM, 0.156 μM, 0.078 μM, 0.039 μM, 0.0195 μM, 0.00975 μM). Affinity data at different concentrations were obtained, and compound K was fitted to the desired concentration. D Numerical value.

[0207] (III) Experimental Results

[0208] The table below shows the K binding of the compounds in the examples to the VISTA protein. D Numerical value.

[0209]

[0210]

[0211] II. Experiment on the binding ability of compounds to VISTA protein

[0212] (I) Experimental Equipment and Reagents

[0213] 1. Centrifuge Eppendorf 5430

[0214] 2. Perkein Elmer EnVision microplate reader

[0215] 3. PD-1-Eu, PD-L1-Biotin, Dye labeled acceptor (BPS Bioscience)

[0216] 4. 384well microplate (Perkin Elmer)

[0217] (II) Experimental Methods

[0218] The inhibitory effect of the compounds in the examples on the PD-1 / PD-L1 pathway was determined using the TR-FRET method, and the procedure is as follows:

[0219] (1) Prepare 1×modified TR-FRET assay buffer;

[0220] (2) Preparation of compound concentration gradients: The test concentration of the test compound was started at 1.0 μM, diluted 3-fold 9 times, for a total of 10 concentration points. The solution was diluted to a final concentration of 100-fold in a 384-well plate, and 200 nmol was transferred to a 384-well plate for reaction. 200 nmol of DMSO was added to each of the negative and positive control wells.

[0221] (3) Prepare PD-L1-Biotin solution with 4 times the final concentration using the 1×modified TR-FRET assay buffer prepared in advance;

[0222] (4) Add 5.0 μL of PD-L1-Biotin solution at 4 times the final concentration to the compound well and the positive control well, respectively; add 5.0 μL of 1×modified TR-FRET assay buffer to the negative control well;

[0223] (5) Centrifuge at 1000 rpm for 0.5 min, shake to mix, and incubate at room temperature for 15 min;

[0224] (6) Prepare a mixed solution of PD-1-Eu with a final concentration of 4 times and Dye-labeled acceptor with a final concentration of 2 times using the pre-prepared 1×modified TR-FRET assay buffer.

[0225] (7) Add 15 μL of a mixed solution of PD-1-Eu and Dye labeled acceptor (containing 5.0 μL of PD-1-Eu at 4 times the final concentration and 10 μL of Dye labeled acceptor at 2 times the final concentration);

[0226] (8) Centrifuge at 1000 rpm for 0.5 min, shake to mix, and incubate at room temperature for 90 min;

[0227] (9) Centrifuge the 384-well plate at 1000 rpm for 0.5 min, shake to mix, and then read the fluorescence intensity at 665 nm and 620 nm using an ELISA reader, and calculate the TR-FRET ratio (665 nm emission / 620 nm emission).

[0228] (10) Calculate the inhibition rate at different concentration points, and use GraphPad Prism 6.0 software to fit the dose-response curve to obtain the IC50 of each compound on the enzyme activity. 50 value.

[0229] (III) Experimental Results

[0230] The table below shows the IC50 values ​​of the compounds in the examples for inhibiting the PD-1 / PD-L1 interaction. 50 value.

[0231] Compound numbering <![CDATA[IC 50 (nM)]]> Compound numbering <![CDATA[IC 50 (nM)]]> 1 112 24 19.1 2 201 25 74.2 3 303 26 78.2 4 20.5 27 83.5 5 15.2 28 24.5 6 10.9 29 10.2 7 25.4 30 12.1 8 37.2 31 21.2 9 43.2 32 36.0 10 79.8 33 21.4 11 63.2 34 92.3 12 56.6 35 73.6 13 31.9 36 26.4 14 6.3 37 80.2 15 9.8 38 57.3 16 25.2 39 37.1 17 18.2 40 204 18 24.5 41 185 19 63.9 42 137 20 15.3 43 235 21 8.4 44 167 22 15.7 45 132 23 22.3

[0232] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, and is not intended to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions made by those skilled in the art to the technical solution of the present invention do not depart from the essence and scope of the technical solution of the present invention.

Claims

1. A compound or a pharmaceutically acceptable salt thereof, which is any one of the following:

2. A pharmaceutical composition comprising a therapeutically effective amount of the compound of claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient and a pharmaceutically acceptable carrier.

3. The pharmaceutical composition according to claim 2, characterized in that, The pharmaceutical composition is selected from capsules, powders, tablets, granules, pills, injections, inhalers, ointments, suppositories, or patches.

4. Use of the compound of claim 1 or a pharmaceutically acceptable salt thereof in the preparation of an inhibitor or antitumor drug having VISTA and PD-1 / PD-L1 signaling pathway inhibitory activity.

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