Double-target small molecule drug and application thereof

Abstract

The invention discloses a compound as shown in a formula I, or pharmaceutically acceptable salt, isomer, metabolite, prodrug, solvate or hydrate, pharmaceutical composition and application thereof. The deuterated isoquinoline-aminopyrimidine compound as shown in the formula I provided by the invention has an obvious effect on blocking VISTA and PD-1 / PD-L1 signal channels, can effectively treat and relieve diseases such as cancers and the like, and is simple to prepare and convenient for industrial production as a small-molecule inhibitor. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the field of chemical medicine technology, and in particular to compounds that block VISTA and PD-1 / PD-L1 signaling pathways, and preparation methods and uses thereof. Background Art

[0002] Malignant tumors are one of the most complex and difficult to treat diseases in the world today, and they seriously threaten human health and life. The main methods of tumor treatment include surgery, radiotherapy, chemotherapy, and targeted therapy. Tumor immunotherapy refers to a treatment method that stimulates the body's immune system to enhance the anti-tumor immune effect, thereby inhibiting and killing tumor cells. With the comprehensive development and cross-penetration of oncology, immunology, and molecular biology, and the in-depth study of the pathogenesis of tumors, immunotherapy has achieved many results, bringing new hope for tumor treatment.

[0003] Immune checkpoint inhibitors in tumor immunotherapy are currently popular immunotherapy drugs. Tumor cells inhibit the activity of immune cells T cells by upregulating the expression of immune checkpoint receptors, thereby completing the immune escape of tumor cells. Immune checkpoint inhibitors inhibit the immune checkpoint pathway, relieve the inhibition of immune cells T cells, activate the body's immune killing of tumor cells, and achieve the effect of tumor treatment. At present, the immune checkpoints that have been discovered include CTLA-4 (cytotoxic T lymphocyte-associated antigen-4), PD-1 (Programmed cell death 1) and TIM3 (T cellmembrane 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, consisting of an immunoglobulin (Ig) superfamily domain, a handle of about 20 amino acids, a transmembrane domain, and an intracellular domain of about 95 amino acid residues, which contains an immunoreceptor tyrosine-based inhibitory motif (ITIM) and an immunoreceptor tyrosine-based switch motif (ITSM). PD-1 is mainly expressed on T cells, B cells, natural killer T cells, activated monocytes, and dendritic cells. PD-1 has two types of natural ligands: PD-L1 and PD-L2; they can interact with PD-1, 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 inhibiting the immune activity of T cells 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 tumors. Currently, a number of monoclonal antibody drugs targeting the PD-1 / PD-L1 signaling pathway have been approved for marketing and are used to treat cancer patients. In addition, small molecule inhibitors of PD-1 / PD-L1 are also being actively developed, and the fastest-progressing ones have entered the clinical phase II stage. However, as the treatment of clinical cancer patients deepens, studies have found that when the PD-1 / PD-L1 pathway is inhibited, it will promote the increased expression of other immune checkpoints, such as VISTA, TIM3, etc., and then lead to drug immune tolerance.

[0005] T cell activation inhibitor immunoglobulin variable region domain (VISTA) is a type of immune checkpoint that is mainly expressed in hematopoietic cell tissues. In addition, VISTA is also highly expressed in bone marrow cells, neuroblasts, and neutrophils. Unlike other immune checkpoints that are induced to express after the activation of the immune response, VISTA is stably expressed during the steady-state period of immune cells. Many studies have shown that VISTA also has an inhibitory effect on the immune system. Therefore, inhibiting the VISTA signaling pathway can also repair the body's anti-tumor immune activity.

[0006] In order to better exert the body's anti-tumor immune function, the study of dual-target small molecule inhibitors will be an effective method. So far, there are no small molecule inhibitors that simultaneously target PD1 / PD-L1 and VISTA signaling pathways on the market. Therefore, it is of great significance to develop new dual-target small molecule inhibitors with good anti-tumor activity. Summary of the invention

[0007] Purpose of the invention: In view of the current situation that there are no dual-target inhibitor drugs for VISTA and PD-1 / PD-L1 signaling pathways on the market, the present invention provides a small molecule compound that simultaneously targets VISTA and PD-1 / PD-L1 signaling pathways, and a preparation method and use thereof.

[0008] Technical solution: In order to achieve the above-mentioned purpose, the present invention discloses a compound shown in the following formula I and a pharmaceutically acceptable salt thereof:

[0009]

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

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

[0012] R2 is independently hydrogen, deuterium, halogen, cyano, morpholinyl, tetrahydropyranyl, substituted or unsubstituted hydroxy, substituted or unsubstituted amino, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted alkoxy, substituted or unsubstituted cycloalkyl, substituted or unsubstituted pyrrolidonyl, substituted or unsubstituted piperidinyl, substituted or unsubstituted piperazinyl;

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

[0014] m is 0, 1, 2 or 3;

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

[0016] Further, the present invention discloses a compound as shown in the following 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, C 1-4 Ester group, C 14 Carboxyl, C 1-4 Hydroxyl; wherein the C 1-4 Alkyl, C 1-4 Amide, C 1-4 Ester group, C 1-4 Carboxyl, C 1-4 The hydroxyl group may be optionally 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 2-4Alkynyl; when there are multiple substituents, the substituents are the same or different.

[0017] Further, the present invention discloses a compound as shown in the following 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 piperazinyl 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, C 1-4 Ester or C 1-4 An amide group; wherein said C 1-4 Alkyl, C 1-4 Amide, C 1-4 Ester group, C 1-4 Carboxyl, C 1-4 The hydroxyl group may be optionally 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 Alkynyl; when there are multiple substituents, the substituents are the same or different.

[0018] The compound represented by formula I of the present application is specifically selected from the following compounds:

[0019]

[0020]

[0021] Furthermore, the above specific compounds also include pharmaceutically acceptable salts, racemates, optical isomers or solvates thereof.

[0022] The preparation method of the above compound, when X is O, NH, S, R1 is a substituted or unsubstituted amino group, and m is 1, the synthesis route of the compound is as follows:

[0023]

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

[0025] (1) Compound II and compound III undergo condensation ring-closing reaction to obtain compound IV;

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

[0027] (3) Compound VI undergoes a nucleophilic substitution reaction to obtain compound VII;

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

[0029] (5) Compound VIII is subjected to reduction reaction to obtain compound IX.

[0030] (6) Compound IX is subjected to oxidation reaction to obtain compound X.

[0031] (7) Compound X is subjected to reductive amination to obtain compound XI.

[0032] A pharmaceutical composition comprising a therapeutically effective amount of one or more compounds having the structure described in general formula I or their pharmaceutically acceptable salts, racemates, optical isomers or solvent compounds as active ingredients and a pharmaceutically acceptable carrier.

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

[0034] The use of the compound 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 activity, anti-tumor drugs, and anti-infective drugs.

[0035] The pharmaceutical composition is used in the preparation of immune checkpoint inhibitors, inhibitors with VISTA and PD-1 / PD-L1 signaling pathway inhibitory activity, anti-tumor drugs, and anti-infective drugs.

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

[0037] The technical scheme of the present invention will be clearly and completely described below in conjunction with specific embodiments. Obviously, the described embodiments are only a part of embodiments of the present invention, rather than all embodiments. Based on the embodiments in the present invention, all other embodiments obtained by those of ordinary skill in the art without making creative work, all belong to 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 common commercial products unless otherwise specified.

[0038] Example 1. Preparation of Compound 1

[0039]

[0040] Synthesis route:

[0041]

[0042] Synthesis of Compound I-2

[0043] The raw 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), refluxed in an oil bath at 80°C for 3 h, and then the reaction solution was cooled to room temperature, the solvent was concentrated, and then dissolved in anhydrous dichloromethane (40 mL), and then 6.04 g of 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ) was added. Under nitrogen protection, the reaction was carried out at room temperature for 1 h. TLC detected that the raw material reaction was complete, 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 make sand, and purified by column chromatography (petroleum ether: ethyl acetate = 20: 1) to obtain compound 2 (7.2 g, 82%).

[0044] Synthesis of Compound I-3

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

[0046] Synthesis of Compound I-4

[0047] Dissolve m-bromophenol (2g, 11.56mmol) in acetonitrile (20mL), add 1-bromo-3-chloropropane (2.18g, 13.87mmol) and potassium carbonate (2.4g, 17.34mmol), and stir for 18h in an oil bath at 55°C. TLC monitoring shows that the raw material has reacted completely. Extract with ethyl acetate, wash with saturated brine, dry with anhydrous sodium sulfate, filter, concentrate the filtrate for sand making, and purify by column chromatography (petroleum ether) to obtain compound 4 (2.5g, 87%).

[0048] Synthesis of Compound I-5

[0049] Compound I-4 (1g, 4.01mmol), compound 3 (1.82g, 4.81mmol), 1,1′-bis(diphenylphosphinoferrocene)palladium dichloride (147mg, 0.2mmol), potassium carbonate (1.11g, 8.02mmol) were dissolved in 1,4-dioxane (15mL) and water (3mL), and a nitrogen balloon was filled for protection, and stirred for 12h in an oil bath at 80°C. TLC detection showed that the raw material reaction was complete, diatomaceous earth was filtered, ethyl acetate was extracted, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated and sanded, and purified by column chromatography (petroleum ether: ethyl acetate = 20: 1) to obtain compound 3 (1.3g, 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 batches under ice bath conditions, and the reaction was carried out at 0°C for 1 h. TLC monitoring showed that the raw material had reacted completely. Methanol was added dropwise to quench the reaction, and the mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to prepare sand, and purified by column chromatography (petroleum ether: ethyl acetate = 2: 1) to obtain 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, and sodium bicarbonate (256 mg, 3.05 mmol) and Dess-Martin reagent (969 mg, 2.29 mmol) were added under ice bath conditions, and then the mixture was moved to room temperature for reaction for 1 h. After the reaction was completed under TLC monitoring, saturated sodium thiosulfate solution and saturated sodium bicarbonate solution were added to quench, and the mixture was extracted with dichloromethane, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to make sand, and purified by column chromatography (petroleum ether: ethyl acetate = 4: 1) to obtain 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 in sequence. The mixture was stirred in an oil bath at 80°C for 12 h. The reaction was completed after monitoring 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 make sand. The mixture 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 stirring was continued at room temperature for 12 h. The reaction was completed after TLC monitoring. Saturated sodium bicarbonate solution was added for washing, and the mixture was extracted with dichloromethane, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to make sand. Compound JC-1 (56 mg, 85%) was obtained after column chromatography (dichloromethane: methanol = 20: 1). 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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 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] Embodiment 46

[0191] Tablet preparation

[0192] Compound 1 (50 g) prepared in Example 1, hydroxypropyl methylcellulose E (150 g), starch (200 g), appropriate amount of povidone K30 and magnesium stearate (1 g) were mixed, granulated and tableted.

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

[0194] Application Examples

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

[0196] 1. Experiment on the binding ability of compounds to VISTA protein

[0197] (I) Experimental equipment and reagents

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

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

[0200] 3. Amino coupling kit. Product number: BR-1000-50, purchased from GE Healthcare.

[0201] 4. Buffer: 10x PBS-P+ (Cat. No. 28-9950-84), purchased from GE Healthcare.

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

[0203] 6. Protein: Glycosylated VISTA protein.

[0204] 7. Other consumables: 1.5 ml EP tube without cap (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 steps

[0206] The binding ability of the compound to VISTA protein was tested using the Biacore S200 system and CM5 chip. The 10 mM compound stock solution 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). The affinity data of different concentrations were obtained by testing, and the compound K was obtained by fitting. D Numeric value.

[0207] (III) Experimental results

[0208] The following table shows the K of the Example compounds binding to the VISTA protein D Numeric value.

[0209]

[0210]

[0211] 2. Compound Binding Ability Experiment with VISTA Protein

[0212] (I) Experimental equipment and reagents

[0213] 1. Centrifuge Eppendorf 5430

[0214] 2. Microplate reader Perkein Elmer EnVision

[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 example compounds on the PD-1 / PD-L1 pathway was determined using the TR-FRET method, and the operating steps are as follows:

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

[0220] (2) Preparation of compound concentration gradient: The test compound concentration is 1.0 μM, 3-fold dilution is performed 9 times, 10 concentration points. Dilute to a solution of 100 times the final concentration in a 384-well plate, transfer 200 nL to the 384-well plate for reaction and set aside. Add 200 nL DMSO to the negative control well and the positive control well respectively;

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

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

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

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

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

[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 and mix, read the fluorescence intensity at 665 nm and 620 nm using a microplate 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-effect curve to calculate the IC of each compound on the enzyme activity. 50 value.

[0229] (III) Experimental results

[0230] The following table shows the IC values ​​of the example compounds for the inhibitory activity of PD-1 / PD-L1 interaction 50 value.

[0231] Compound No. <![CDATA[IC 50 (nM)]]> Compound No. <![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, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate 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 of the following:

2. A pharmaceutical composition comprising a therapeutically effective amount of the compound according to 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, inhalants, ointments, suppositories or patches.

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

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