Indoline compound containing pyrazole structure as well as preparation method and application of indoline compound

Abstract

The invention discloses an indoline compound containing a pyrazole structure as well as a preparation method and application thereof, belongs to the technical field of medicines, and relates to the indoline compound containing the pyrazole structure as shown in a general formula I, a stereoisomer of the indoline compound and pharmaceutically acceptable salt of the indoline compound. The invention also relates to a preparation method of the compounds and application of the compounds in preparation of drugs for treating diseases related to PD-1 / PD-L1 protein / protein interaction. The compound provided by the invention has high-level inhibition activity on PD-1 / PD-L1 protein / protein interaction, so that the compound has a good inhibition effect on PD-1 / PD-L1 protein / protein interaction. The indoline compound containing the pyrazole structure, the stereoisomer of the indoline compound and the pharmaceutically acceptable salt of the indoline compound can be used for preparing drugs for diseases related to PD-1 / PD-L1 signal channel abnormity, such as cancers, infectious diseases and autoimmune diseases.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to an indoline compound containing a pyrazole structure, its preparation method, and its application. Background Technology

[0002] Immunotherapy-based cancer treatments have made significant progress in the past decade, and their rapid development has fundamentally changed the landscape of clinical cancer treatment. Among these, programmed death 1 (PD-1) and programmed cell death-ligand 1 (PD-L1) are currently the most prominent immune checkpoints. PD-L1 regulates T cell function by interacting with PD-1, inhibiting T cell activation, proliferation, and survival, playing a crucial role in autoimmunity and immune responses. Tumor cells, by abnormally overexpressing PD-L1, construct an immunosuppressive tumor microenvironment, thereby achieving immune escape. Blocking the PD-1 / PD-L1 interaction can restore the function of T cells attacking tumor cells and restore the body's anti-tumor immune response, opening up new avenues for cancer treatment. Monoclonal antibodies targeting PD-1 / PD-L1 have shown significant efficacy in clinical practice, and more than ten monoclonal antibody drugs have been approved for marketing, with indications including non-small cell lung cancer, urothelial carcinoma, melanoma, and head and neck cancer.

[0003] While monoclonal antibody drugs possess unique advantages such as high selectivity and significant clinical efficacy, their use is limited to some extent by drawbacks, including susceptibility to protease degradation, requiring administration only by injection, poor oral bioavailability, poor tumor tissue penetration, and severe immune-related adverse reactions. Compared to monoclonal antibodies, small molecule inhibitors show greater promise in these aspects and are expected to provide a potential alternative to monoclonal antibody drugs. Summary of the Invention

[0004] To address the problems existing in the prior art, this invention designs and synthesizes a series of novel indoline compounds containing pyrazole structures. Studies have shown that these compounds can significantly inhibit PD-1 / PD-L1 protein / protein interactions and have important potential application value.

[0005] This invention relates to indoline compounds containing a pyrazole structure as shown in general formula I, their stereoisomers, and pharmaceutically acceptable salts.

[0006]

[0007] in,

[0008] R1 is selected from

[0009] R2 is selected from hydrogen or (C1-C4) alkyl;

[0010] R3 and R4 are each independently selected from hydrogen, (C1-C4)alkyl, (C3-C8)cycloalkyl, hydroxy(C1-C4)alkyl, amino(C1-C4)alkyl, carbamoyl(C1-C4)alkyl, aminosulfonyl(C1-C4)alkyl, methanesulfonylamino(C1-C4)alkyl, aminosulfonylamino(C1-C4)alkyl, carboxyl(C1-C4)alkyl, (C1-C4)alkoxyformyl(C1-C4)alkyl, and 4-7 membered heterocyclic alkyl groups containing 1-3 N, O, or S atoms; In this context, the (C1-C4)alkyl, (C3-C8)cycloalkyl, hydroxy(C1-C4)alkyl, amino(C1-C4)alkyl, carbamoyl(C1-C4)alkyl, aminosulfonyl(C1-C4)alkyl, methanesulfonylamino(C1-C4)alkyl, aminosulfonylamino(C1-C4)alkyl, carboxyl(C1-C4)alkyl, and (C1-C4)alkoxyformyl(C1-C4)alkyl groups may contain 1-3 4-7 membered heterocyclic alkyl groups selected from N, O, and S atoms, which may optionally be substituted by 1-3 R9 atoms;

[0011] Alternatively, R3, R4, and the nitrogen atoms bonded to them can form a 4-6 member nitrogen-containing heterocycle; the nitrogen-containing heterocycle contains 1-3 heteroatoms selected from N, S, and O; the nitrogen-containing heterocycle may optionally be bonded by 1-3 R atoms. 10 replace;

[0012] R5 is selected from hydrogen, (C1-C4) alkyl, and halogen;

[0013] R6 is selected from hydrogen and (C1-C4)alkoxy groups, wherein the (C1-C4)alkoxy group can be... replace;

[0014] R7 and R8 are each independently selected from hydrogen and (C1-C4) alkyl groups;

[0015] Alternatively, R7, R8, and the nitrogen atoms bonded to them can form a 4-6 member nitrogen-containing heterocycle; the nitrogen-containing heterocycle contains 1-3 heteroatoms selected from N, S, and O; the nitrogen-containing heterocycle may optionally be bonded by 1-3 R atoms. 11 replace;

[0016] R9 is selected from hydrogen, halogen, hydroxyl, carboxyl, amino, (C1-C4)alkyl, hydroxy(C1-C4)alkyl, amino(C1-C4)alkyl, (C1-C4)alkoxy(C1-C4)alkyl, and (C1-C4)acyl.

[0017] R 10 Selected from hydrogen, hydroxyl, carboxyl, (C1-C4)alkyl, and hydroxy(C1-C4)alkyl;

[0018] R 11 Selected from hydrogen, hydroxyl, carboxyl, (C1-C4)alkyl, and hydroxy(C1-C4)alkyl.

[0019] This invention preferably relates to indoline compounds containing a pyrazole structure as shown in general formula I, their stereoisomers, and pharmaceutically acceptable salts, wherein:

[0020] R1 is selected from

[0021] R2 is a (C1-C4) alkyl group;

[0022] R3 and R4 are each independently selected from hydrogen, (C1-C4)alkyl, hydroxy(C1-C4)alkyl, amino(C1-C4)alkyl, carbamoyl(C1-C4)alkyl, carboxyl(C1-C4)alkyl, and 4-7 membered heterocyclic alkyl containing 1-3 N, O, or S atoms; wherein, the (C1-C4)alkyl, hydroxy(C1-C4)alkyl, amino(C1-C4)alkyl, carbamoyl(C1-C4)alkyl, carboxyl(C1-C4)alkyl, and 4-7 membered heterocyclic alkyl containing 1-3 N, O, or S atoms may optionally be replaced by 1-3 R9 atoms;

[0023] Alternatively, R3, R4, and the nitrogen atoms bonded to them can form a 4-6 member nitrogen-containing heterocycle; the nitrogen-containing heterocycle contains 1-3 heteroatoms selected from N, S, and O; the nitrogen-containing heterocycle may optionally be bonded by 1-3 R atoms. 10 replace;

[0024] R5 is selected from hydrogen, (C1-C4) alkyl, and halogen;

[0025] R6 is selected from hydrogen and (C1-C4)alkoxy groups, wherein the (C1-C4)alkoxy group can be... replace;

[0026] R7 and R8 are each independently (C1-C4) alkyl groups;

[0027] Alternatively, R7, R8, and the nitrogen atoms bonded to them can form a 4-6 member nitrogen-containing heterocycle; the nitrogen-containing heterocycle contains 1-3 heteroatoms selected from N, S, and O; the nitrogen-containing heterocycle may optionally be bonded by 1-3 R atoms. 11 replace;

[0028] R9 is selected from hydrogen, halogen, hydroxyl, carboxyl, amino, (C1-C4)alkyl, hydroxy(C1-C4)alkyl, amino(C1-C4)alkyl, (C1-C4)alkoxy(C1-C4)alkyl, and (C1-C4)acyl.

[0029] R 10Selected from hydrogen, hydroxyl, carboxyl, (C1-C4)alkyl, and hydroxy(C1-C4)alkyl;

[0030] R 11 It is selected from hydrogen, hydroxyl, carboxyl, and (C1-C4) alkyl groups.

[0031] The present invention more preferably relates to indoline compounds containing a pyrazole structure as shown in general formula I, their stereoisomers, and pharmaceutically acceptable salts, wherein:

[0032] R1 is selected from

[0033] R2 is a methyl group;

[0034] R3, R4, and the nitrogen atom bonded to them, as well as the methylene moiety bonded to the nitrogen atom: Selected from the following structures:

[0035]

[0036] R5 is selected from fluorine, chlorine, and bromine;

[0037] R7, R8, and the nitrogen atoms attached to them: Selected from

[0038] The indoline compounds containing pyrazole structures represented by general formula I of this invention, their stereoisomers, and pharmaceutically acceptable salts are preferably selected from the following compounds, but these compounds do not imply any limitation on this invention:

[0039]

[0040] Furthermore, the present invention also includes prodrugs of the compounds of the present invention. The prodrugs of the compounds of the present invention are derivatives of general formula I, which may have weak or no activity on their own, but are converted into the corresponding biologically active form under physiological conditions (e.g., through metabolism, solvation or other means) after administration.

[0041] The above-described indoline compounds of general formula I containing a pyrazole structure, their stereoisomers, and pharmaceutically acceptable salts, wherein the pharmaceutically acceptable salts include salts formed with inorganic acids, organic acids, and alkali metal ions; the inorganic acids are selected from: hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, and phosphoric acid; the organic acids are selected from: succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, methanesulfonic acid, ethanesulfonic acid, and p-toluenesulfonic acid; and the alkali metal ions are selected from lithium ions, sodium ions, and potassium ions.

[0042] In this invention, "halogen" refers to fluorine, chlorine, bromine, or iodide; "alkyl" refers to a straight-chain or branched alkyl group; and "nitrogen-containing heterocycle" refers to a monocyclic or polycyclic cyclic system containing nitrogen atoms, wherein the cyclic system is non-aromatic or aromatic. This represents the junction of substituents.

[0043] This invention can prepare a composition by mixing an indoline compound of general formula I containing a pyrazole structure, or its stereoisomer, or a pharmaceutically acceptable salt thereof, as an active ingredient, with a pharmaceutically acceptable carrier or excipient. The carrier or excipient includes diluents, binders, wetting agents, disintegrants, lubricants, and flow aids known in the art. Diluents include, but are not limited to, starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, and dicalcium phosphate; wetting agents include, but are not limited to, water, ethanol, and isopropanol; binders include, but are not limited to, starch paste, dextrin, syrup, honey, glucose solution, gum arabic paste, gelatin paste, sodium carboxymethyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, and polyethylene glycol; disintegrants include, but are not limited to, dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, croscarmellose, croscarmellose sodium, sodium carboxymethyl starch, and sodium dodecyl sulfonate; lubricants and flow aids include, but are not limited to, talc, silica, and polyethylene glycol.

[0044] The pharmaceutical composition of the present invention can be formulated into several dosage forms, including but not limited to injections, tablets, capsules, etc.

[0045] The pyrazole-containing indoline compounds and their stereoisomers, as well as pharmaceutically acceptable salts of the present invention, can be used in combination with other active ingredients to achieve better therapeutic effects.

[0046] This invention also provides the use of indoline compounds of general formula I containing a pyrazole structure, their stereoisomers, and pharmaceutically acceptable salts in the preparation of medicaments for the prevention and / or treatment of diseases related to abnormalities in the PD-1 / PD-L1 signaling pathway. The diseases related to abnormalities in the PD-1 / PD-L1 signaling pathway include cancer, infectious diseases, and autoimmune diseases. The cancers include lymphoma, non-small cell lung cancer, small cell lung cancer, head and neck cell carcinoma, glioma, neuroblastoma, squamous cell carcinoma of the lung, adenocarcinoma of the lung, bladder cancer, gastric cancer, colon cancer, colorectal cancer, kidney cancer, bile duct cancer, gastric cancer, esophageal squamous cell carcinoma, ovarian cancer, pancreatic cancer, breast cancer, prostate cancer, liver cancer, brain cancer, melanoma, multiple myeloma, skin cancer, epithelial cell carcinoma, leukemia, and cervical cancer; the infectious diseases include bacterial infections and viral infections; and the autoimmune diseases include organ-specific autoimmune diseases and systemic autoimmune diseases. The organ-specific autoimmune diseases include chronic lymphocytic thyroiditis, hyperthyroidism, insulin-dependent diabetes mellitus, ulcerative colitis, and acute idiopathic polyneuritis. The systemic autoimmune diseases include rheumatoid arthritis, systemic lupus erythematosus, systemic vasculitis, and autoimmune hemolytic anemia.

[0047] The positive and progressive effects of this invention are as follows: the indoline compounds containing pyrazole structures of this invention have novel chemical structures, most of which have shown significant inhibitory activity against PD-1 / PD-L1 interaction in in vitro biological activity studies, and the compounds can be used for the treatment and prevention of various diseases such as cancer. Detailed Implementation

[0048] The following synthetic routes summarize and describe the preparation of indoline compounds of general formula I containing pyrazole structures according to the present invention. All starting materials were prepared by methods well known to those skilled in the art of organic chemistry, or are commercially available, as described in these procedures. The compounds of the present invention are prepared by methods described in these procedures or by similar methods well known to those skilled in the art of organic chemistry.

[0049] Route 1:

[0050]

[0051] (a) Intermediate 2 was prepared from benzyl acetone 1 and diethyl oxalate as raw materials under the action of sodium ethoxide;

[0052] (b) Intermediate 3 was prepared by using intermediate 2 and alkyl hydrazine sulfate as raw materials in the presence of potassium carbonate;

[0053] (c) Using intermediate 3 as raw material, intermediate 4 was prepared by oxidation using a potassium osmium tetroxide / sodium periodate oxidation system;

[0054] (d) Intermediate 5 was prepared from intermediate 4 and ethylene glycol under the action of p-toluenesulfonic acid monohydrate.

[0055] (e) Intermediate 6 was prepared by ester hydrolysis using intermediate 5 as a raw material;

[0056] (f) Using intermediate 6 and 4-substituted indoline as raw materials, intermediate 7 was prepared by amidation reaction under condensing agent conditions;

[0057] (g) Using intermediate 7 as a raw material, intermediate 8 was prepared by deprotection of p-toluenesulfonic acid monohydrate;

[0058] (h) Using intermediate 8 and amine compounds as raw materials, the compound shown in general formula I is prepared by reductive amination reaction in the presence of sodium triacetoxyborohydride or sodium cyanoborohydride.

[0059] Route 2:

[0060]

[0061] (a) Intermediate 10 was prepared by Suzuki-Miyaura coupling reaction using 4-bromoindole 9 and pinacol diboronic acid ester as raw materials.

[0062] (b) Intermediate 11 was prepared by the Suzuki-Miyaura reaction using intermediate 10 and halophenol as raw materials;

[0063] (c) Using intermediate 11 as raw material and sodium cyanoborohydride as reducing agent, intermediate 12 is prepared by reduction reaction;

[0064] (d) Using intermediate 12 and intermediate 6 as raw materials, intermediate 13 was prepared by amidation reaction under condensation agent conditions;

[0065] (e) Using intermediate 13 and haloalkane as raw materials, intermediate 14 was prepared by nucleophilic substitution under the action of potassium carbonate;

[0066] (f) Using intermediate 14 as a raw material, intermediate 15 was prepared by deprotection of p-toluenesulfonic acid monohydrate;

[0067] (g) Using intermediate 15 and an amine compound as raw materials, R1 is prepared by reductive amination reaction under the action of sodium triacetoxyborohydride. The compound represented by general formula I at that time.

[0068] The definitions of R1, R2, R3, R4, R5, and R6 are as described above. All indoline compounds of general formula I containing a pyrazole structure can be prepared by the method described above or a similar method.

[0069] The following non-limiting embodiments are intended to enable those skilled in the art to more fully understand the invention, but do not limit the invention in any way.

[0070] Example 1: (5-{[(2-hydroxyethyl)amino]methyl}-1-methyl-1H-pyrazol-3-yl)(4-phenylindoline-1-yl)methyl ketone (Compound I-1)

[0071]

[0072] Step 1: 4-Phenyl-1H-indole

[0073]

[0074] At room temperature, 4-bromo-1H-indole (9.05 g, 46.42 mmol), phenylboronic acid (7.50 g, 61.51 mmol), Pd(dppf)Cl2 (1.01 g, 1.39 mmol), and potassium carbonate (21.51 g, 155.63 mmol) were successively added to a mixed solution of 1,4-dioxane (80 mL) and water (20 mL), and reacted at 80 °C for 4 h under N2 protection. After the reaction was complete, the reaction solution was cooled to room temperature, filtered through a diatomaceous earth sieve, the filter cake was washed with ethyl acetate, the filtrate was concentrated, water was added and extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated and purified by column chromatography to obtain 7.62 g of white solid, with a yield of 85.0%. 1 H NMR (600MHz, DMSO-d6) δ11.27(s,1H),7.67(d,J=7.3Hz,2H),7.49(t,J=7.7Hz,2H),7.42(t,J=5.9Hz,2H) ,7.37(t,J=7.4Hz,1H),7.18(t,J=7.6Hz,1H),7.08(d,J=7.1Hz,1H),6.55(s,1H).ESI-MSm / z:194.2[M+H] + .

[0075] Step 2: 4-Phenylindoline

[0076]

[0077] 4-Phenyl-1H-indole (7.62 g, 39.46 mmol) was added to 50 mL of glacial acetic acid at room temperature. Then, sodium cyanoborohydride (9.92 g, 157.85 mmol) was slowly added under ice bath conditions. After the addition was complete, the reaction mixture was allowed to react at room temperature for 3 h. After the reaction was complete, the reaction solution was poured into 200 mL of water, and the pH was adjusted to 9-10 with 40% NaOH solution under ice bath conditions. The mixture was extracted with ethyl acetate (200 mL × 3), and the organic layer was washed with saturated brine. The solution was concentrated under reduced pressure, and column chromatography was used to separate 6.76 g of a white solid, with a yield of 88.4%. 1 H NMR (600MHz, DMSO-d6) δ7.47–7.40(m,4H),7.36–7.29(m,1H),7.00(t,J=7.7Hz,1H),6.59(d,J=7.1Hz,1H) ,6.51(d,J=7.7Hz,1H),5.59(s,1H),3.39(t,J=8.4Hz,2H),2.97(t,J=8.4Hz,2H).ESI-MSm / z:196.1[M+H] + .

[0078] Step 3: Ethyl 2,4-dioxo-6-phenylhex-5-enoate

[0079]

[0080] Sodium ethoxide (0.93 g, 13.67 mmol) was dissolved in 150 mL of anhydrous diethyl ether. Then, under ice bath conditions, 50 mL of a mixture of benzylidene acetone (20 g, 136.81 mmol) and diethyl oxalate (24 mL, 177.36 mmol) in diethyl ether was added. After the addition was complete, the reaction was allowed to proceed at room temperature for 4 h. The reaction was monitored by TLC until complete. The mixture was filtered, and the filter cake was washed with diethyl ether and dried at room temperature for 1 h. The filter cake was dissolved in 600 mL of dichloromethane, and 200 mL of 1N sulfuric acid was added. The mixture was stirred for 15 min, and the organic layer was separated. The organic layer was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 27.27 g of an orange solid, with a yield of 81.0%. 1 H NMR (600MHz, CDCl3) δ7.68(d,J=15.9Hz,1H),7.51(s,1H),7.35(s,1H),6.60(d,J=15.9Hz, 1H),6.48(s,1H),4.31(dd,J=14.1,7.0Hz,1H),1.49–1.18(m,1H).ESI-MSm / z:269.3[M+Na] + .

[0081] Step 4: Ethyl 1-methyl-5-styryl-1H-pyrazole-3-carboxylate

[0082]

[0083] Ethyl 2,4-dioxo-6-phenylhex-5-enoate (27.84 g, 108.62 mmol) was dissolved in 300 mL of ethanol at room temperature. Potassium carbonate (18.75 g, 135.66 mmol) and methylhydrazine sulfate (19.56 g, 135.69 mmol) were added. After the addition was complete, the reaction was carried out at 80 °C for 3 h. The reaction was monitored by TLC until complete. The mixture was filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography to give 16.45 g of an orange oil, with a yield of 86.5%. 1 H NMR (600MHz, CDCl3) δ7.43 (d, J = 7.5Hz, 1H), 7.32 (t, J = 7.6Hz, 1H), 7.25 (t, J = 7.3Hz, 1H), 7.02 (d, J = 16. 2Hz, 1H), 6.95 (s, 1H), 6.83 (d, J = 16.2Hz, 1H), 4.35 (q, J = 7.1Hz, 1H), 3.94 (s, 1H), 1.34 (t, J = 7.1Hz, 1H).

[0084] Step 5: Ethyl 5-formyl-1-methyl-1H-pyrazole-3-carboxylate

[0085]

[0086] Ethyl 1-methyl-5-styryl-1H-pyrazole-3-carboxylate (16.25 g, 63.45 mmol) was dissolved in 500 mL of a mixed solution of dioxane and water (V / V, 4:1). K₂O₄·2H₂O (1.18 g, 3.81 mmol) was added, and the mixture was stirred at room temperature for 30 min. Then, NaIO₄ (54.76 g, 256.02 mmol) was added, and the reaction was allowed to proceed for 1 h at room temperature. The reaction was monitored by TLC until complete. The mixture was filtered, and the filtrate was extracted with 600 mL of water and ethyl acetate (200 mL × 3). The organic layers were combined. The organic layers were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography to give 9.02 g of a white solid (70.4% yield). 1 H NMR (600MHz, CDCl3) δ9.89(s,1H),7.42(s,1H),4.44(q,J=7.1Hz,2H),4.26(s,3H),1.42(t,J=7.1Hz,3H).ESI-MS m / z:183.0[M+H] + .

[0087] Step 6: Ethyl 5-(1,3-dioxolane-2-yl)-1-methyl-1H-pyrazole-3-carboxylate

[0088]

[0089] Ethyl 5-formyl-1-methyl-1H-pyrazole-3-carboxylate (7.95 g, 43.66 mol) was dissolved in 56 mL of toluene, and ethylene glycol (4.8 mL, 85.84 mmol) and p-toluenesulfonic acid monohydrate (0.67 g, 3.49 mmol) were added. The mixture was heated to reflux and reacted for 6 h. The reaction was monitored by TLC until complete. After the reaction solution was cooled to room temperature, it was washed successively with saturated sodium carbonate solution and saturated brine. The organic layer was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 9.62 g of a pale yellow oil, with a yield of 97.5%. 1 H NMR (600MHz, CDCl3) δ6.80 (s, 1H), 5.90 (s, 1H), 4.32 (q, J = 7.1Hz, 2H), 4.05–3.96 (m, 5H), 3.92 (s, 3H), 1.32 (t, J = 7.1Hz, 3H).

[0090] Step 7: 5-(1,3-dioxolane-2-yl)-1-methyl-1H-pyrazole-3-carboxylic acid

[0091]

[0092] Ethyl 5-(1,3-dioxolane-2-yl)-1-methyl-1H-pyrazole-3-carboxylate (9.62 g, 42.55 mmol) was dissolved in 30 mL of a mixed solution of tetrahydrofuran and methanol (V / V, 4:1) at room temperature. 20 mL of 2N NaOH aqueous solution was added, and the reaction was allowed to proceed for 30 min at room temperature. The reaction was confirmed to be complete by TLC. The reaction solution was concentrated under reduced pressure, and the residue was diluted with an appropriate amount of water. The pH was adjusted to 1-2 by slowly adding 12N concentrated HCl at 0 °C. The mixture was filtered, and the filter cake was washed with a small amount of dichloromethanol and dried to give 7.87 g of a pale yellow solid, with a yield of 93.4%.

[0093] Step 8: [5-(1,3-dioxolane-2-yl)-1-methyl-1H-pyrazol-3-yl](4-phenylindoline-1-yl)methyl ketone

[0094]

[0095] 5-(1,3-dioxolane-2-yl)-1-methyl-1H-pyrazole-3-carboxylic acid (2.0 g, 10.10 mmol), 4-phenylindoline (1.97 g, 10.09 mmol), and HATU (4.61 g, 12.12 mmol) were added to 20 mL of DMF and stirred at room temperature for 15 min. DIPEA (5.22 g, 40.40 mmol) was then added to the reaction mixture, and the reaction was continued at room temperature for 2 h. The reaction was monitored by TLC until complete. The reaction mixture was poured into 150 mL of ice water, filtered, and the filter cake was dried to give 6.7 g of a pale yellow solid, with a yield of 70.6%. 1 H NMR(600MHz,DMSO-d6)δ8.21(s,1H),7.51–7.45(m,4H),7.40–7.37(m,1H),7.31(t,J=7.7Hz,1H),7.10(d,J=7.6Hz,1H),6.83(s,1H),6.09 (s,1H),4.48(s,2H),4.09(dd,J=8.7,5.3Hz,2H),4.00(dd,J=8.7,5.2Hz,2H),3.92(s,3H),3.21(t,J=8.3Hz,2H).ESI-MSm / z:376.4[M+H] + .

[0096] Step 9: 1-Methyl-3-(4-phenylindoline-1-carbonyl)-1H-pyrazole-5-carboxaldehyde

[0097]

[0098] [5-(1,3-dioxolane-2-yl)-1-methyl-1H-pyrazol-3-yl](4-phenylindololin-1-yl) methyl ketone (4.19 g, 11.17 mmol) was dissolved in 20 mL of a mixture of acetone and water (V / V, 4:1), and p-toluenesulfonic acid monohydrate (4.25 g, 22.34 mmol) was added. The reaction was carried out at 70 °C for 1 h. The reaction was monitored by TLC until complete. The reaction solution was poured into 20 mL of ice water, filtered, and the filter cake was purified by column chromatography to give 2.88 g of white solid, yield 77.9%. 1 H NMR(600MHz, CDCl3)δ9.85(s,1H),8.29(s,1H),7.46(s,1H),7.39–7.34(m,4H),7.31 –7.25(m,2H),7.05(d,J=7.5Hz,1H),4.51(s,2H),4.16(s,3H),3.17(t,J=8.2Hz,2H).

[0099] Step 10: (5-{[(2-hydroxyethyl)amino]methyl}-1-methyl-1H-pyrazol-3-yl)(4-phenylindoline-1-yl)methyl ketone

[0100]

[0101] 1-Methyl-3-(4-phenylindoline-1-carbonyl)-1H-pyrazole-5-carboxaldehyde (100 mg, 0.3 mmol) was dissolved in 3 mL of dichloromethane. Ethanolamine (55 mg, 0.9 mmol) and one drop of glacial acetic acid were added, and the mixture was stirred at room temperature for 2 h. Sodium cyanoborohydride (57 mg, 0.9 mmol) was then added, and the mixture was stirred at room temperature for 4 h. The reaction was monitored by TLC until complete, and the reaction solution was washed with water. The organic layer was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography to give 57.8 mg of a white solid (51.2% yield). 1 H NMR(600MHz,DMSO-d6)δ8.21(s,1H),7.51–7.44(m,4H),7.41–7.35(m,1H),7.32–7.27(m,1H),7.08(d,J=7.6Hz,1H),6.71(s,1H), 4.58 (s, 1H), 4.49 (t, J = 8.3Hz, 2H), 3.89 (s, 3H), 3.85 (s, 2H), 3.49 (t, J = 5.7Hz, 2H), 3.20 (t, J = 8.3Hz, 2H), 2.65 (t, J = 5.7Hz, 2H).

[0102] Following the synthetic method in step 10 of Example 1, 1-methyl-3-(4-phenylindoline-1-carbonyl)-1H-pyrazole-5-carboxaldehyde was used as a starting material and reductively amination was performed with the corresponding small molecule amines to obtain the target compounds in Examples 2-5 and 12-15.

[0103] Example 2: N-[2-({[1-methyl-3-(4-phenylindoline-1-carbonyl)-1H-pyrazol-5-yl]methyl}amino)ethyl]acetamide (compound I-2)

[0104]

[0105] 1H NMR(600MHz,DMSO-d6)δ8.21(s,1H),7.85–7.78(m,1H),7.51–7.44(m,4H),7.41–7.35(m,1H),7.33–7.28(m,1H),7.08(d,J=7.7Hz,1H),6.6 9(s,1H),4.49(t,J=8.2Hz,2H),3.88(s,3H),3.79(s,2H),3.20(t,J=8.3Hz,2H),3.14(q,J=6.3Hz,2H),2.58(t,J=6.5Hz,2H),1.80(s,3H).

[0106] Example 3: (1-Methyl-5-{[(tetrahydro-2H-pyran-4-yl)amino]methyl}-1H-pyrazol-3-yl)(4-phenylindoline-1-yl)methyl ketone (compound I-3)

[0107]

[0108] 1 H NMR (600MHz, DMSO-d6) δ8.21(s,1H),7.51–7.44(m,4H),7.42–7.35(m,1H),7.33–7.27(m,1H),7.08(d,J=7.6Hz,1H),6.69(s,1H),4.49(t,J=7.8H z,2H),3.89(s,3H),3.82(d,J=4.1Hz,4H),3.28(td,J=11.5,2.2Hz,2H), 3.20(t,J=8.3Hz,2H),2.67–2.60(m,1H),1.82(d,2H),1.33–1.21(m,2H).

[0109] Example 4: (5-{[(1-hydroxy-2-methylpropen-2-yl)amino]methyl}-1-methyl-1H-pyrazol-3-yl)(4-phenylindoline-1-yl)methyl ketone (compound I-4)

[0110]

[0111] 1H NMR(600MHz,DMSO-d6)δ8.21(s,1H),7.52–7.44(m,4H),7.42–7.36(m,1H),7.34–7.28(m,1H),7.09(d,J=7.6Hz,1H),6. 80(s,1H),4.49(t,J=8.3Hz,2H),3.92(s,3H),3.38(d,J=14.1Hz,2H),3.21(t,J=8.3Hz,2H),1.91(s,2H),1.14(s,6H).

[0112] Example 5: {5-[(3-hydroxyazacyclobutane-1-yl)methyl]-1-methyl-1H-pyrazol-3-yl}(4-phenylindoline-1-yl)methyl ketone (compound I-5)

[0113]

[0114] 1 H NMR (600MHz, DMSO-d6) δ8.21(s,1H),7.51–7.44(m,4H),7.42–7.35(m,1H),7.3–7.27(m,1H),7.08(d,J=7.6Hz,1H),6.65(s,1H),5.36(d,J=6. 3Hz,1H),4.47(t,J=8.3Hz,2H),4.20(h,J=6.2Hz,1H),3.85(s,3H),3.65(s,2H),3.54–3.49(m,2H),3.20(t,J=8.3Hz,2H),2.86–2.81(m,2H).

[0115] Example 6: (R)-{5-[(3-hydroxypyrrolidone-1-yl)methyl]-1-methyl-1H-pyrazole-3-yl}(4-phenylindoline-1-yl)methyl ketone (compound I-12)

[0116]

[0117] 1H NMR (600MHz, DMSO-d6) δ8.21(s,1H),7.48(q,J=8.0Hz,4H),7.38(t,J=6.8Hz,1H),7.30(t,J=7.6Hz,1 H),7.09(d,J=7.6Hz,1H),6.67(s,1H),4.72(d,J=4.4Hz,1H),4.49(s,2H),3.89(s,3H),3.66(dd,J=33 .0,13.9Hz,2H),3.37(dt,J=20.9,10.5Hz,1H),3.20(t,J=8.2Hz,2H),2.66(ddd,J=22.9,12.3,7.0Hz ,2H),2.40(ddd,J=12.8,11.7,5.6Hz,2H),2.00(dt,J=14.2,7.4Hz,1H),1.56(dt,J=12.8,8.1Hz,1H).

[0118] Example 7: (S)-{5-[(3-hydroxypyrrolidone-1-yl)methyl]-1-methyl-1H-pyrazole-3-yl}(4-phenylindoline-1-yl)methyl ketone (compound I-13)

[0119]

[0120] 1 H NMR (600MHz, DMSO-d6) δ8.21 (s, 1H), 7.54–7.42 (m, 4H), 7.39 (dd, J = 9.2, 4.4Hz, 1H), 7.30 ( t,J=7.6Hz,1H),7.09(d,J=7.6Hz,1H),6.66(s,1H),4.72(s,1H),4.49(s,2H),3.89(s,3H), 3.66(dd,J=33.1,13.9Hz,2H),3.33(s,1H),3.20(t,J=8.2Hz,2H),2.66(ddd,J=23.1,12.4 ,7.0Hz,2H),2.47–2.31(m,2H),2.00(dt,J=14.3,7.5Hz,1H),1.56(dt,J=12.9,8.1Hz,1H).

[0121] Example 8: (R)-1-{[1-methyl-3-(4-phenylindoline-1-carbonyl)-1H-pyrazole-5-yl]methyl}pyrrolidine-3-carboxylic acid (compound I-14)

[0122]

[0123] 1H NMR(600MHz,DMSO-d6)δ8.21(s,1H),7.51–7.44(m,4H),7.41–7.36(m,1H),7.30(t ,J=7.7Hz,1H),7.08(d,J=7.6Hz,1H),6.67(s,1H),4.49(s,2H),3.88(s,3H),3.68 (q,J=13.9Hz,2H),3.20(t,J=8.3Hz,2H),2.93(dt,J=14.8,7.5Hz,1H),2.71(t,J= 8.7Hz, 1H), 2.68–2.62 (m, 1H), 2.53 (t, J = 6.9Hz, 2H), 1.97 (dd, J = 14.4, 7.2Hz, 2H).

[0124] Example 9: (S)-1-{[1-methyl-3-(4-phenylindoline-1-carbonyl)-1H-pyrazole-5-yl]methyl}pyrrolidine-3-carboxylic acid (Compound I-15)

[0125]

[0126] 1 H NMR (600MHz, DMSO-d6) δ8.21(s,1H),7.48(q,J=8.0Hz,4H),7.38(t,J=6.8Hz,1H ),7.30(t,J=7.5Hz,1H),7.08(d,J=7.6Hz,1H),6.68(s,1H),4.49(s,2H),3.88(s ,3H),3.68(q,J=13.9Hz,2H),3.20(t,J=8.2Hz,2H),2.94(dt,J=15.0,7.6Hz,1H ), 2.69(dt,J=15.3,9.0Hz,2H),2.53(dd,J=13.5,6.6Hz,2H),2.04–1.92(m,2H).

[0127] Example 10: {[1-methyl-3-(4-phenylindoline-1-carbonyl)-1H-pyrazol-5-yl]methyl}glycine (Compound I-6)

[0128]

[0129] Step 1: {[1-methyl-3-(4-phenylindoline-1-carbonyl)-1H-pyrazol-5-yl]methyl}glycine ethyl ester

[0130]

[0131] Using 1-methyl-3-(4-phenylindoline-1-carbonyl)-1H-pyrazole-5-carboxaldehyde prepared in step 9 of Example 1 as a raw material and glycine ethyl ester as a raw material, a light yellow oily substance was prepared by reduction amination according to the synthesis method in step 10 of Example 1, with a yield of 75.4%. 1 H NMR(600MHz,DMSO-d6)δ8.21(s,1H),7.51–7.45(m,4H),7.41–7.36(m,1H),7.30(t,J=7.8Hz,1H),7.08(d,J=7.6Hz,1H),6.66 (s,1H),4.49(t,J=7.2Hz,2H),3.89(s,3H),3.81(s,2H),3.63(s,3H),3.36(d,J=12.4Hz,2H),3.33(s,3H),3.23–3.15(m,2H).

[0132] Step 2: {[1-methyl-3-(4-phenylindoline-1-carbonyl)-1H-pyrazol-5-yl]methyl}glycine

[0133]

[0134] At room temperature, ethyl glycine {[1-methyl-3-(4-phenylindoline-1-carbonyl)-1H-pyrazol-5-yl]methyl} (100 mg, 0.24 mmol) and LiOH·H₂O (100 mg, 2.38 mmol) were dissolved in a mixed solution of methanol and water (V / V, 3:1), and reacted at 50 °C for 3 h. The reaction was confirmed to be complete by TLC. The reaction solution was diluted with water, and the pH was adjusted to 1-2 by slow dropwise addition of 12N concentrated HCl at 0 °C. The solution was filtered and dried to give 74.6 mg of a pale yellow solid, with a yield of 80%. 1 H NMR(600MHz,DMSO-d6)δ8.23(s,1H),7.51–7.45(m,4H),7.41–7.37(m,1H),7.32(t,J=7.6Hz,1H ),7.12–7.07(m,2H),4.49(s,2H),4.37(s,2H),4.01(s,3H),3.96(s,2H),3.22(t,J=8.3Hz,2H).

[0135] Following the synthesis method in step 1 of Example 10, 1-methyl-3-(4-phenylindoline-1-carbonyl)-1H-pyrazole-5-carboxaldehyde was used as a raw material and reduced-amination was performed with the corresponding amino acid ethyl ester. Then, the target compounds in Examples 7-11 were obtained by ester hydrolysis in step 2.

[0136] Example 11: {[1-methyl-3-(4-phenylindoline-1-carbonyl)-1H-pyrazol-5-yl]methyl}-D-alanine (Compound I-7)

[0137]

[0138] 1 H NMR (600MHz, DMSO-d6) δ8.23(s,1H),7.48(q,J=8.0Hz,4H),7.39(t,J=6.7Hz,1H),7.32(t,J=7.5Hz,1H),7.13–7.03(m,2 H), 4.49 (s, 2H), 4.37 (q, J = 14.4Hz, 2H), 4.15 (q, J = 6.9Hz, 1H), 4.01 (s, 3H), 3.22 (t, J = 8.2Hz, 2H), 1.58 (d, J = 7.1Hz, 3H).

[0139] Example 12: {[1-methyl-3-(4-phenylindoline-1-carbonyl)-1H-pyrazol-5-yl]methyl}-L-alanine (Compound I-8)

[0140]

[0141] 1 H NMR (600MHz, DMSO-d6) δ8.22(s,1H),7.53–7.44(m,4H),7.42–7.36(m,1H),7.32(t,J=7.6Hz,1H),7.10(d,J=7.6Hz,1H),7. 00(s,1H),4.49(s,2H),4.35–4.26(m,2H),4.01(d,J=7.3Hz,1H),3.99(s,3H),3.22(t,J=8.2Hz,2H),1.52(d,J=7.1Hz,3H).

[0142] Example 13: {[1-methyl-3-(4-phenylindoline-1-carbonyl)-1H-pyrazol-5-yl]methyl}-D-serine (Compound I-9)

[0143]

[0144] 1H NMR(600MHz,DMSO-d6)δ8.21(s,1H),7.59–7.45(m,4H),7.43–7.34(m,1H),7. 27(t,J=7.5Hz,1H),7.10(d,J=7.5Hz,1H),7.09(s,1H),5.75(d,J=25.9Hz,1H) ,4.47(s,2H),4.39(q,J=14.5Hz,2H),4.17(d,J=3.1Hz,1H),4.06(dd,J=12.0 ,2.7Hz,1H),4.00(s,3H),3.90(dd,J=12.0,3.7Hz,1H),3.21(t,J=8.3Hz,2H).

[0145] Example 14: {[1-methyl-3-(4-phenylindoline-1-carbonyl)-1H-pyrazol-5-yl]methyl}-L-serine (Compound I-10)

[0146]

[0147] 1 H NMR(600MHz,DMSO-d6)δ8.22(s,1H),7.52–7.45(m,4H),7.41–7.37(m,1H),7. 32(t,J=7.6Hz,1H),7.11(d,J=7.6Hz,1H),7.06(s,1H),5.74(d,J=25.9Hz,1H) ,4.49(s,2H),4.40(q,J=14.5Hz,2H),4.17(d,J=3.1Hz,1H),4.06(dd,J=12.0 ,2.7Hz,1H),3.99(s,3H),3.96(dd,J=12.0,3.6Hz,1H),3.23(t,J=8.3Hz,2H).

[0148] Example 15: 2-Methyl-2-({[1-methyl-3-(4-phenylindoline-1-carbonyl)-1H-pyrazol-5-yl]methyl}amino)propionic acid (compound I-11)

[0149]

[0150] 1H NMR(600MHz, DMSO-d6)δ8.23(s,1H),7.52–7.46(m,4H),7.39(dd,J=9.4,4.3Hz,1H),7.32(t,J=7.7Hz,1H),7 .10(d,J=7.6Hz,1H),7.08(s,1H),4.50(s,2H),4.32(s,2H),4.03(s,3H),3.23(t,J=8.3Hz,2H),1.64(s,6H).

[0151] Example 16: [4-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)indoline-1-yl](5-{[(2-hydroxyethyl)amino]methyl}-1-methyl-1H-pyrazol-3-yl)methyl ketone (compound I-16)

[0152]

[0153] Step 1: 1,4-Benzodioxane-6-boronate pinacol ester

[0154]

[0155] At room temperature, 6-bromo-1,4-benzodioxane (10 g, 46.7 mmol), pinacol diborate (14.3 g, 56.1 mmol), Pd(dppf)Cl2·CH2Cl2 (1.15 g, 1.4 mmol), and potassium acetate (11.5 g, 116.8 mmol) were added sequentially to anhydrous 1,4-dioxane, and the mixture was reacted at 90 °C for 3 h under N2 protection. After the reaction was complete, the reaction solution was cooled to room temperature, concentrated, and 200 mL of ethyl acetate was added. The mixture was filtered through a diatomaceous earth liner, the filter cake was washed with ethyl acetate, and the concentrated filtrate was purified by column chromatography to obtain 11.9 g of a white solid, with a yield of 97.4%.

[0156] Step 2: 4-(2,3-dihydro-1,4-benzodioxane-6-yl)indoline

[0157]

[0158] Similar to the synthesis method of 4-phenylindoline in Example 1, the intermediate was prepared from 1,4-benzodioxane-6-borate pinacol ester and 4-bromo-1H-indole via a Suzuki-Miyaura coupling reaction and a Gribble indole reduction reaction, with an overall yield of 74.7%. ESI-MS m / z: 254.2 [M+H] + .

[0159] Step 3: 5-{[4-(2,3-dihydrobenzo[2,1-b][1,4]dioxane-6-yl)-2,3-dihydro-1H-indole-1-yl]carbonyl}-2-methylpyrazol-3-carboxaldehyde

[0160]

[0161] The compound was prepared as a white solid using 4-(2,3-dihydro-1,4-benzodioxane-6-yl)indoline and 5-(1,3-dioxolane-2-yl)-1-methyl-1H-pyrazole-3-carboxylic acid as starting materials, following the same synthetic method as steps 8-9 in Example 1. 1 H NMR (600MHz, CDCl3) δ9.84(s,1H),8.25(s,1H),7.45(s,1H),7.24(s,1H),7.01(d,J=7.5Hz,1H), 6.85(ddd,J=10.2,7.2,1.9Hz,3H),4.50(s,2H),4.23(s,4H),4.16(s,3H),3.17(t,J=8.2Hz,2H).

[0162] Step 4: [4-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)indoline-1-yl](5-{[(2-hydroxyethyl)amino]methyl}-1-methyl-1H-pyrazol-3-yl)methyl ketone (compound I-16)

[0163]

[0164] The compound was prepared from ethanolamine using the same synthetic method as step 10 in Example 1. It was a white solid with a yield of 49.3%. 1 H NMR (600MHz, DMSO-d6) δ8.16(s,1H),7.25(t,J=8.0Hz,1H),7.03(d,J=7.7Hz,1H),6.96(s,1H),6.93(s,2H),6.68(s,1H),4.4 8(t,J=8.2Hz,2H),4.28(s,4H),3.88(s,3H),3.81(s,2H),3.48(t,J=5.7Hz,2H),3.19(t,J=8.3Hz,2H),2.62(t,J=5.7Hz,2H).

[0165] Following the synthesis method in step 4 of Example 16, the target compounds in Examples 17-20 were prepared by reductive amination of 5-{[4-(2,3-dihydrobenzo[2,1-b][1,4]dioxane-6-yl)-2,3-dihydro-1H-indol-1-yl]carbonyl}-2-methylpyrazol-3-carboxaldehyde with the corresponding small molecule amines.

[0166] Example 17: N-{2-[({3-[4-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)indoline-1-carbonyl]-1-methyl-1H-pyrazol-5-yl}methyl)amino]ethyl}acetamide (Compound I-17)

[0167]

[0168] 1 H NMR (600MHz, DMSO-d6) δ8.16(s,1H),7.86–7.80(m,1H),7.28–7.23(m,1H),7.03(d,J=7.6Hz,1H),6.97(s,1H),6.93(s,2H),6. 69(s,1H),4.47(t,J=8.3Hz,2H),4.28(s,4H),3.88(s,3H),3.80(s,2H),3.22–3.12(m,4H),2.59(t,J=6.5Hz,2H),1.80(s,3H).

[0169] Example 18: [4-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)indoline-1-yl](1-methyl-5-{[(tetrahydro-2H-pyran-4-yl)amino]methyl}-1H-pyrazol-3-yl) methyl ketone (compound I-18)

[0170]

[0171] 1 H NMR(600MHz, DMSO-d6)δ8.16(s,1H),7.29–7.22(m,1H),7.03(d,J=7.7Hz,1H),6.98–6.91(m,3H),6.69(s,1H),4.47(t,J=8.2Hz,2H),4.28 (s,4H),3.89(s,3H),3.86–3.80(m,4H),3.28(td,J=11.5,2.2Hz,2H),3.19(t,J=8.3Hz,2H),2.65(s,1H),1.82(d,2H),1.34–1.25(m,2H).

[0172] Example 19: [4-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)indoline-1-yl](5-{[(1-hydroxy-2-methylpropen-2-yl)amino]methyl}-1-methyl-1H-pyrazol-3-yl)methyl ketone (compound I-19)

[0173]

[0174] 1 H NMR(600MHz,DMSO-d6)δ8.16(s,1H),7.28–7.23(m,1H),7.03(d,J=7.7Hz,1H),6.96(s,1H),6.93(s,2H),6.69(s,1H ), 4.72 (s, 1H), 4.47 (t, 2H), 4.28 (s, 4H), 3.90 (s, 3H), 3.79 (s, 2H), 3.33 (s, 2H), 3.19 (t, J = 8.3Hz, 2H), 1.05 (s, 6H).

[0175] Example 20: [4-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)indoline-1-yl]{5-[(3-hydroxyazacyclobutane-1-yl)methyl]-1-methyl-1H-pyrazole-3-yl} methyl ketone (Compound I-20)

[0176]

[0177] 1 H NMR(600MHz,DMSO-d6)δ8.16(s,1H),7.29–7.22(m,1H),7.03(d,J=7.6Hz,1H),6.96(s,1H),6.93(s,2H),6.65(s,1H),5.37(d,J=6.3Hz,1H),4.4 6(t,J=8.1Hz,2H),4.28(s,4H),4.21(h,J=6.3Hz,1H),3.85(s,3H),3.66 (s,2H),3.53(t,J=6.8Hz,2H),3.18(t,J=8.3Hz,2H),2.88–2.83(m,2H).

[0178] Example 21: [4-(2-fluoro-3-{[3-(tetrahydro-1H-pyrrolo-1-yl)propyl]oxy}phenyl)-2,3-dihydro-1H-indol-1-yl](5-{[(2-hydroxyethyl)amino]methyl}-1-methylpyrazol-3-yl) methyl ketone (compound I-21)

[0179]

[0180] Step 1: 4-(4,4,5,5-Tetramethyl-1,3,2-dioxaborhexacyclopentan-2-yl)-1H-indole

[0181]

[0182] At room temperature, 25 g (127.5 mmol) of 4-bromoindole, 38.9 g (153.0 mmol) of pinacol diborate, 1.9 g (2.6 mmol) of Pd(dppf)Cl2, and 37.5 g (382.5 mmol) of potassium acetate were added sequentially to 80 mL of 1,4-dioxane. The reaction was carried out at 100 °C for 3 h under N2 protection. After the reaction was completed, the reaction solution was cooled to room temperature, filtered through a diatomaceous earth filter, the filter cake was washed with ethyl acetate, the filtrate was concentrated, water was added and extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, evaporated to dryness, and slurried with diethyl ether to give 25.8 g of white solid, with a yield of 83.2%.

[0183] Step 2: 2-Fluoro-3-(1H-indol-4-yl)phenol

[0184]

[0185] At room temperature, 10.7 g (44.0 mmol) of 4-(4,4,5,5-tetramethyl-1,3,2-dioxaboranecyclopentan-2-yl)-1H-indole, 10.9 g (57.1 mmol) of 3-bromo-2-fluorophenol, 1.0 g (1.32 mmol) of Pd(dppf)Cl2, and 21.4 g (154.8 mmol) of potassium carbonate were added sequentially to 100 mL of a mixed solution of 1,4-dioxane and water (V / V, 3:1). The reaction was carried out at 90 °C for 4 h under N2 protection. After the reaction was completed, the reaction solution was cooled to room temperature, filtered through a diatomaceous earth sieve, the filter cake was washed with ethyl acetate, the filtrate was concentrated, water was added and extracted with ethyl acetate, the organic layer was dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated and purified by column chromatography to obtain 8.2 g of white solid, with a yield of 82.5%.

[0186] Step 3: 3-(2,3-dihydro-1H-indole-4-yl)-2-fluorophenol

[0187]

[0188] 2-Fluoro-3-(1H-indol-4-yl)phenol (3 g, 13.1 mmol) was added to 10 mL of glacial acetic acid at room temperature. Then, sodium cyanoborohydride (2.5 g, 39.3 mmol) was slowly added under ice bath conditions. After the addition was complete, the reaction mixture was allowed to react at room temperature for 3 h. Once the reaction was complete, the reaction solution was poured into 200 mL of water, and the pH was adjusted to 9-10 with 40% NaOH solution under ice bath conditions. The mixture was extracted with ethyl acetate (100 mL × 3), and the organic layer was washed with saturated brine. The solution was concentrated under reduced pressure, and column chromatography was used to separate 2.5 g of a white solid, with a yield of 83.2%.

[0189] Step 4: [5-(1,3-dioxacyclopentan-2-yl)-1-methylpyrazol-3-yl][4-(2-fluoro-3-hydroxyphenyl)-2,3-dihydro-1H-indole-1-yl] methyl ketone

[0190]

[0191] The 5-(1,3-dioxolane-2-yl)-1-methyl-1H-pyrazole-3-carboxylic acid (2 g, 10.1 mmol) and 3-(2,3-dihydro-1H-indol-4-yl)-2-fluorophenol (2.3 g, 10.1 mmol) prepared in step 7 of Example 1 were dissolved in 50 mL of DCM. EDCI (2.5 g, 13.1 mmol) was added under cold trap, and the reaction was carried out at room temperature for 30 min. After the reaction was detected by TLC, the reaction solution was added to water, extracted with DCM (20 mL × 3), the organic layer was washed with saturated brine, concentrated under reduced pressure, and purified by column chromatography to obtain 3.15 g of a pale yellow solid, with a yield of 76.4%.

[0192] Step 5: [5-(1,3-dioxacyclopent-2-yl)-1-methylpyrazol-3-yl][4-(2-fluoro-3-{[3-(tetrahydro-1H-pyrrolo-1-yl)propyl]oxy}phenyl)-2,3-dihydro-1H-indol-1-yl] methyl ketone

[0193]

[0194] [5-(1,3-dioxacyclopent-2-yl)-1-methylpyrazole-3-yl][4-(2-fluoro-3-hydroxyphenyl)-2,3-dihydro-1H-indol-1-yl] methyl ketone (3.9 g, 9.5 mmol), 1-(3-chloropropyl)pyrrolidine (1.67 g, 11.4 mmol), and K₂CO₃ (3.3 g, 23.75 mmol) were dissolved in 30 mL of DMF and reacted at 70 °C for 3 h. The reaction mixture was poured into water and extracted with EA (20 mL × 3). The organic layer was washed with saturated brine and dried under evaporation. 3.9 g of a pale yellow oil was obtained, with a yield of 79.1%.

[0195] Step 6: 5-{[4-(2-fluoro-3-{[3-(tetrahydro-1H-pyrrolo-1-yl)propyl]oxy}phenyl)-2,3-dihydro-1H-indol-1-yl]carbonyl}-2-methylpyrazol-3-carboxaldehyde

[0196]

[0197] [5-(1,3-dioxacyclopent-2-yl)-1-methylpyrazol-3-yl][4-(2-fluoro-3-{[3-(tetrahydro-1H-pyrrolo-1-yl)propyl]oxy}phenyl)-2,3-dihydro-1H-indol-1-yl] methyl ketone (3 g, 5.6 mmol) was dissolved in 30 mL of a mixture of acetone and water (V / V, 4:1), and p-toluenesulfonic acid monohydrate (2.13 g, 11.2 mmol) was added. The reaction was carried out at 70 °C for 1 h. The pH was adjusted to weakly alkaline with 2 M hydrochloric acid, and the mixture was extracted with DCM (20 mL × 3). The organic layer was washed with saturated brine, concentrated under reduced pressure, and purified by column chromatography to give 2.1 g of a colorless oil, with a yield of 76.3%.

[0198] Step 7: [4-(2-fluoro-3-{[3-(tetrahydro-1H-pyrrolo-1-yl)propyl]oxy}phenyl)-2,3-dihydro-1H-indol-1-yl](5-{[(2-hydroxyethyl)amino]methyl}-1-methylpyrazol-3-yl) methyl ketone (compound I-21)

[0199]

[0200] 1 H NMR(400MHz, DMSO-d6)δ8.21(s,1H),7.29(t,J=7.9Hz,1H),7.23–7.15(m,2H),7.00(d,J=7 .6Hz,1H),6.92(dt,J=9.2,3.9Hz,1H),6.68(s,1H),4.55–4.45(m,2H),4.13(t,J=6.4Hz,2H ),3.88(s,3H),3.79(s,2H),3.47(d,J=6.5Hz,2H),3.01(t,J=8.3Hz,2H),2.60(t,J=5.8Hz, 2H),2.55(t,J=7.1Hz,2H),2.43(d,J=5.8Hz,4H),1.92(p,J=6.8Hz,2H),1.71–1.64(m,4H).

[0201] 5-{[4-(2-fluoro-3-{[3-(tetrahydro-1H-pyrrolo-1-yl)propyl]oxy}phenyl)-2,3-dihydro-1H-indol-1-yl]carbonyl}-2-methylpyrazol-3-carboxaldehyde (100 mg, 0.2 mmol) was dissolved in 3 mL of dichloromethane. Ethanolamine (30.6 mg, 0.6 mmol) and one drop of glacial acetic acid were added, and the mixture was stirred at room temperature for 2 h. Sodium cyanoborohydride (38 mg, 0.6 mmol) was then added, and the mixture was stirred at room temperature for 4 h. The reaction was monitored by TLC until complete, and the reaction mixture was washed with water. The organic layer was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography to give 63.4 mg of a white solid, with a yield of 60.8%.

[0202] Following the synthetic method in step 7 of Example 21, the target compounds in Examples 22-24 were prepared by reductive amination of 5-{[4-(2-fluoro-3-{[3-(tetrahydro-1H-pyrrolo-1-yl)propyl]oxy}phenyl)-2,3-dihydro-1H-indol-1-yl]carbonyl}-2-methylpyrazol-3-carboxaldehyde or 5-{[4-(2-chloro-3-{[3-(tetrahydro-1H-pyrrolo-1-yl)propyl]oxy}phenyl)-2,3-dihydro-1H-indol-1-yl]carbonyl}-2-methylpyrazol-3-carboxaldehyde with the corresponding small molecule amines.

[0203] Example 22: [4-(2-fluoro-3-{[3-(tetrahydro-1H-pyrrolo-1-yl)propyl]oxy}phenyl)-2,3-dihydro-1H-indol-1-yl](5-{[(1-hydroxy-2-methylpropyl-2-yl)amino]methyl}-1-methylpyrazol-3-yl) methyl ketone (Compound I-22)

[0204]

[0205] 1 H NMR (400MHz, DMSO-d6) δ8.21 (s, 1H), 7.23 (dd, J = 37.2, 7.3Hz, 4H), 7.05–6. 85(m,2H),6.66(s,1H),4.63(s,1H),4.48(t,J=8.4Hz,2H),4.12(d,J=6.9H z,2H),3.88(s,3H),3.73(s,2H),3.25(s,2H),2.99(d,J=8.7Hz,2H),2.57( d,J=6.8Hz,2H),1.98–1.91(m,2H),1.69(s,4H),1.24(s,4H),1.02(s,6H).

[0206] Example 23: [4-(2-chloro-3-{[3-(tetrahydro-1H-pyrrolo-1-yl)propyl]oxy}phenyl)-2,3-dihydro-1H-indol-1-yl](5-{[(2-hydroxyethyl)amino]methyl}-1-methylpyrazol-3-yl) methyl ketone (compound I-23)

[0207]

[0208] 1 H NMR(400MHz, DMSO-d6)δ8.15(s,1H),7.30(t,J=7.9Hz,1H),7.23(t,J=7.9Hz,1H),7 .12(d,J=8.3Hz,1H),6.86(dd,J=7.7,4.0Hz,2H),6.63(s,1H),4.48–4.42(m,2H),4. 09(t,J=6.3Hz,2H),3.82(s,3H),3.74(s,2H),2.85(t,J=8.4Hz,2H),2.54(q,J=7.7, 6.9Hz, 4H), 2.39 (d, J = 5.9Hz, 6H), 1.90 (q, J = 6.7Hz, 2H), 1.64 (q, J = 3.9, 3.4Hz, 4H).

[0209] Example 24: [4-(2-chloro-3-{[3-(tetrahydro-1H-pyrrolo-1-yl)propyl]oxy}phenyl)-2,3-dihydro-1H-indol-1-yl](5-{[(1-hydroxy-2-methylpropyl-2-yl)amino]methyl}-1-methylpyrazol-3-yl) methyl ketone (compound I-24)

[0210]

[0211] 1 H NMR(400MHz, DMSO-d6)δ8.19(s,1H),7.25(d,J=8.4Hz,1H),7.23(d,J=6.1Hz,2H), 7.06(d,J=7.7Hz,1H),6.95(d,J=7.9Hz,1H),6.69(s,1H),4.64(s,1H),4.45(t,J= 8.4Hz,2H),4.11(d,J=6.9Hz,2H),3.87(s,3H),3.74(s,2H),2.96(d,J=8.7Hz,2H) ,2.55(d,J=6.8Hz,2H),2.02–1.93(m,2H),1.68(s,4H),1.25(s,4H),1.05(s,6H).

[0212] Pharmacological activity study of the compounds of this invention

[0213] 1. Evaluation of the inhibitory activity of the compounds in the examples on the PD-1 / PD-L1 interaction

[0214] Homogenous time-resolved fluorescence (HTRF) assays were used to detect the ability of the compounds of this invention to inhibit PD-1 / PD-L1 interactions. The detection kit (CAT#64PD1PEG) contained the necessary reagents for the experiment, including Anti-Tag1 Eu Cyptate, Anti-Tag2 XL665, Tag1-PD-L1, Tag2-PD-1, and Detection Buffer.

[0215] Experimental steps:

[0216] (1) First, dilute the compound to be tested to 1 mM with DMSO to obtain a stock solution. Then, continue to dilute the stock solution with DMSO at a 4-fold ratio to obtain 6-8 concentrations. Then, dilute the compound solutions of each concentration 20 times with detection buffer to obtain the working solutions of the compound at each concentration.

[0217] (2) Dilute the Tag2-PD-1 and Tag1-PD-L1 stock solutions 40 times with Detection Buffer.

[0218] (3) Add 2 μL of compound working solution, 4 μL of Tag2-PD-1 and 4 μL of Tag1-PD-L1 solution to a 96-well plate in sequence, mix thoroughly and incubate at room temperature for 15 min.

[0219] (4) Dilute Anti-Tag1 Eu Cyptate and Anti-Tag2 XL665 stock solutions 50 times with Detection buffer, mix them in equal volumes, and then add 10 μL of the mixture to each reaction well. Seal the membrane and incubate at room temperature for 1 h.

[0220] (5) The fluorescence signal was detected using a SpectraMax i3X multi-functional microplate reader (320nm excitation, 665nm and 620nm emission).

[0221] 2. The results of the compounds' activity in inhibiting PD-1 / PD-L1 interaction are shown in Table 1.

[0222] Table 1. Activity tests of compounds in the examples to inhibit PD-1 / PD-L1 interaction.

[0223]

[0224]

[0225] Experimental results show that the compounds in the examples can significantly inhibit PD-1 / PD-L1 protein / protein interactions at the biochemical level, and have potential immunomodulatory effects.

Claims

1. Indoline compounds containing a pyrazole structure as shown in Formula I, or their stereoisomers or pharmaceutically acceptable salts thereof; in, R1 is selected from R2 is selected from hydrogen or (C1-C4) alkyl; R3 and R4 are each independently selected from hydrogen, (C1-C4)alkyl, (C3-C8)cycloalkyl, hydroxy(C1-C4)alkyl, amino(C1-C4)alkyl, carbamoyl(C1-C4)alkyl, aminosulfonyl(C1-C4)alkyl, methanesulfonylamino(C1-C4)alkyl, aminosulfonylamino(C1-C4)alkyl, carboxyl(C1-C4)alkyl, (C1-C4)alkoxyformyl(C1-C4)alkyl, and 4-7 membered heterocyclic alkyl groups containing 1-3 N, O, or S atoms; In this context, the (C1-C4)alkyl, (C3-C8)cycloalkyl, hydroxy(C1-C4)alkyl, amino(C1-C4)alkyl, carbamoyl(C1-C4)alkyl, aminosulfonyl(C1-C4)alkyl, methanesulfonylamino(C1-C4)alkyl, aminosulfonylamino(C1-C4)alkyl, carboxyl(C1-C4)alkyl, and (C1-C4)alkoxyformyl(C1-C4)alkyl groups may contain 1-3 4-7 membered heterocyclic alkyl groups selected from N, O, and S atoms, which may optionally be substituted by 1-3 R9 atoms; Alternatively, R3, R4, and the nitrogen atoms bonded to them can form a 4-6 member nitrogen-containing heterocycle; the nitrogen-containing heterocycle contains 1-3 heteroatoms selected from N, S, and O; the nitrogen-containing heterocycle may optionally be bonded by 1-3 R atoms. 10 replace; R5 is selected from hydrogen, (C1-C4) alkyl, and halogen; R6 is selected from hydrogen and (C1-C4)alkoxy groups, wherein the (C1-C4)alkoxy group can be... replace; R7 and R8 are each independently selected from hydrogen and (C1-C4) alkyl groups; Alternatively, R7, R8, and the nitrogen atoms bonded to them can form a 4-6 member nitrogen-containing heterocycle; the nitrogen-containing heterocycle contains 1-3 heteroatoms selected from N, S, and O; the nitrogen-containing heterocycle may optionally be bonded by 1-3 R atoms. 11 replace; R9 is selected from hydrogen, halogen, hydroxyl, carboxyl, amino, (C1-C4)alkyl, hydroxy(C1-C4)alkyl, amino(C1-C4)alkyl, (C1-C4)alkoxy(C1-C4)alkyl, and (C1-C4)acyl. R 10 Selected from hydrogen, hydroxyl, carboxyl, (C1-C4)alkyl, and hydroxy(C1-C4)alkyl; R 11 Selected from hydrogen, hydroxyl, carboxyl, (C1-C4)alkyl, and hydroxy(C1-C4)alkyl.

2. The indoline compound containing a pyrazole structure according to claim 1, or its stereoisomer or a pharmaceutically acceptable salt thereof, characterized in that, In general formula I: R1 is selected from R2 is a (C1-C4) alkyl group; R3 and R4 are each independently selected from hydrogen, (C1-C4)alkyl, hydroxy(C1-C4)alkyl, amino(C1-C4)alkyl, carbamoyl(C1-C4)alkyl, carboxyl(C1-C4)alkyl, and 4-7 membered heterocyclic alkyl containing 1-3 N, O, or S atoms; wherein, the (C1-C4)alkyl, hydroxy(C1-C4)alkyl, amino(C1-C4)alkyl, carbamoyl(C1-C4)alkyl, carboxyl(C1-C4)alkyl, and 4-7 membered heterocyclic alkyl containing 1-3 N, O, or S atoms may optionally be replaced by 1-3 R9 atoms; Alternatively, R3, R4, and the nitrogen atoms bonded to them can form a 4-6 member nitrogen-containing heterocycle; the nitrogen-containing heterocycle contains 1-3 heteroatoms selected from N, S, and O; the nitrogen-containing heterocycle may optionally be bonded by 1-3 R atoms. 10 replace; R5 is selected from hydrogen, (C1-C4) alkyl, and halogen; R6 is selected from hydrogen and (C1-C4)alkoxy groups, wherein the (C1-C4)alkoxy group can be... replace; R7 and R8 are each independently (C1-C4) alkyl groups; Alternatively, R7, R8, and the nitrogen atoms bonded to them can form a 4-6 member nitrogen-containing heterocycle; the nitrogen-containing heterocycle contains 1-3 heteroatoms selected from N, S, and O; the nitrogen-containing heterocycle may optionally be bonded by 1-3 R atoms. 11 replace; R9 is selected from hydrogen, halogen, hydroxyl, carboxyl, amino, (C1-C4)alkyl, hydroxy(C1-C4)alkyl, amino(C1-C4)alkyl, (C1-C4)alkoxy(C1-C4)alkyl, and (C1-C4)acyl. R 10 Selected from hydrogen, hydroxyl, carboxyl, (C1-C4)alkyl, and hydroxy(C1-C4)alkyl; R 11 It is selected from hydrogen, hydroxyl, carboxyl, and (C1-C4) alkyl groups.

3. The indoline compound containing a pyrazole structure according to claim 2, or its stereoisomer or a pharmaceutically acceptable salt thereof, characterized in that, in: R1 is selected from R2 is a methyl group; R3, R4, and the nitrogen atom bonded to them, as well as the methylene moiety bonded to the nitrogen atom: Selected from the following structures: R5 is selected from fluorine, chlorine, and bromine; R7, R8, and the nitrogen atoms attached to them: Selected from 4. The indoline compound containing a pyrazole structure according to claim 3, or its stereoisomer or a pharmaceutically acceptable salt thereof, characterized in that, Preferably from the following compounds:

5. A pharmaceutical composition, characterized in that, The active ingredient comprises an indoline compound with a pyrazole structure as described in any one of claims 1-4, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient, in dosage forms including injections, tablets, and capsules.

6. The use of the indoline compound containing a pyrazole structure as described in any one of claims 1-4, or its stereoisomer or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition as described in claim 5, in the preparation of a medicament for the prevention and / or treatment of diseases related to abnormalities of the PD-1 / PD-L1 signaling pathway.

7. The application according to claim 6, characterized in that, The diseases associated with abnormal PD-1 / PD-L1 signaling pathways include cancer, infectious diseases, and autoimmune diseases.

8. The application according to claim 7, characterized in that, The cancers mentioned include lymphoma, non-small cell lung cancer, small cell lung cancer, head and neck cell carcinoma, glioma, neuroblastoma, squamous cell carcinoma of the lung, adenocarcinoma of the lung, bladder cancer, gastric cancer, colon cancer, colorectal cancer, kidney cancer, bile duct cancer, gastric cancer, esophageal squamous cell carcinoma, ovarian cancer, pancreatic cancer, breast cancer, prostate cancer, liver cancer, brain cancer, melanoma, multiple myeloma, skin cancer, epithelial cell carcinoma, leukemia, and cervical cancer; the infectious diseases mentioned include bacterial infections and viral infections; the autoimmune diseases mentioned include organ-specific autoimmune diseases and systemic autoimmune diseases.

9. The application according to claim 8, characterized in that, The organ-specific autoimmune diseases include chronic lymphocytic thyroiditis, hyperthyroidism, insulin-dependent diabetes mellitus, ulcerative colitis, and acute idiopathic polyneuritis. The systemic autoimmune diseases include rheumatoid arthritis, systemic lupus erythematosus, systemic vasculitis, and autoimmune hemolytic anemia.

Images