A nitrogen-containing heterocyclic compound

Abstract

The present invention discloses a novel IRAK4 inhibitor compound represented by formula (I) and uses of the compound for preventing or treating IRAK4-mediated diseases, including but not limited to autoimmune diseases such as rheumatoid arthritis and lupus erythematosus, activated B-cell-like diffuse large B-cell lymphoma, chronic lymphocytic leukemia, relapsed non-Hodgkin's lymphoma and other lymphomas.

Description

Technical Field

[0001] The present invention relates to a novel IRAK4 inhibitor compound and use of the compound in preventing or treating IRAK4-mediated diseases. Background Art

[0002] Chronic inflammatory and autoimmune diseases threaten the lives of millions of people worldwide and are associated with abnormal signaling mediated by inflammation. Furthermore, some diseases, such as type 2 diabetes and cardiovascular disease, are also associated with abnormal signaling mediated by inflammation.

[0003] Interleukin-1 receptor-associated kinase 4 (IRAK4) is a member of the IRAK family of intracellular serine-threonine kinases. Other members of this kinase family include IRAK2, IRAK-M, and IRAK3. Upon ligand binding to the interleukin-1 receptor (IL-1R) or Toll-like receptor (TLR), IRAK4 mediates signal transduction and activates the expression of downstream inflammatory factors. TLRs receive ligand signals generated by interactions with microorganisms or stimulation by endogenous substances, as well as the initial wave of inflammatory and innate immune responses triggered by these stimuli. TLRs play a crucial role in many diseases, including infections and autoinflammatory disorders, as well as many other human diseases. Like tumor necrosis factor-α (TNF-α) and other major cytokines, interleukin-1 (IL-1) is a key player in inflammatory pathways, capable of propagating and amplifying signals. Since the signaling pathways mediated by TLR, IL-1R and other cytokine receptors have a cross-linking effect, IRAK4, a key signaling factor downstream of the TLR and IL-1R inflammatory pathways, plays an important role in systemic inflammatory responses and can serve as an effective potential target for the treatment of various inflammation-related diseases. Summary of the Invention

[0004] The present invention provides a novel class of IRAK4 inhibitor compounds, their uses, and pharmaceutical compositions containing the same. The compounds have a better inhibitory effect on IRAK4 kinase.

[0005] The present invention provides a compound represented by formula (I) or a pharmaceutically acceptable salt thereof:

[0006]

[0007] Where R1 is C 1-3 Alkyl, R2 is halogen or H;

[0008] Preferably, R1 is methyl or ethyl, and R2 is H or F.

[0009] The compound represented by formula (I) or a pharmaceutically acceptable salt thereof may be the following compound:

[0010]

[0011] The present invention also provides use of the above compound or a pharmaceutically acceptable salt thereof in the preparation of an IRAK4 inhibitor.

[0012] The present invention also provides use of the above compound or a pharmaceutically acceptable salt thereof in preparing a medicament for preventing or treating a disease mediated by abnormal expression of IRAK4.

[0013] The “diseases mediated by abnormal expression of IRAK4” include but are not limited to autoimmune diseases such as rheumatoid arthritis, lupus erythematosus, activated B-cell-like diffuse large B-cell lymphoma, chronic lymphocytic leukemia, relapsed non-Hodgkin's lymphoma and other lymphomas.

[0014] The present invention also provides a pharmaceutical composition comprising the above compound or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0015] As used herein, the term "pharmaceutically acceptable salt" refers to a pharmaceutically acceptable organic or inorganic salt of a compound of the present invention.

[0016] As used herein, the term "pharmaceutically acceptable" means that the substance or composition must be chemically and / or toxicologically compatible with the other ingredients comprising the formulation and / or the mammal to be treated therewith.

[0017] Certain compounds of the present invention possess chiral carbon atoms, double bonds, carbocycles, or heterocycles. The resulting configurational isomers {optical isomers (e.g., enantiomers, diastereomers, etc.), cis-trans isomers, etc.} and mixtures thereof (e.g., racemates) are all within the scope of protection of the present invention. When certain compounds of the present invention indicate a specific configuration, this refers to the presence of that configurational isomer and the absence of substantially all other isomers.

[0018] As used herein, the term "inhibitor" refers to a molecule that reduces, blocks, prevents, delays activation, inactivates, desensitizes, or downregulates, for example, a gene, protein, ligand, receptor, or cell.

[0019] As used herein, the term "abnormal expression of IRAK4" refers to an increase in IRAK4 activity caused by signaling through IRAK4. DETAILED DESCRIPTION

[0020] The present invention is further illustrated by way of examples below, but the present invention is not limited to the scope of the examples. Experimental methods in the following examples where specific conditions are not specified were performed according to conventional methods and conditions, or selected according to the product specifications.

[0021] Example 1

[0022]

[0023] A solution of (7R,7aS)-7-ethyl-3,3-dimethyltetrahydropyrrolo[1,2-c]oxazol-5(1H)-one (300 mg, 1.73 mmol) and Lawesson's reagent (350 mg, 0.86 mmol) in toluene (10 mL) was heated to reflux for 16 hours. The reaction solution was cooled to room temperature and concentrated. The residue was purified by flash chromatography (ethyl acetate / petroleum ether = 0-50%) to obtain 0002A (100 mg, 29% as a yellow solid). LCMS (ESI) [M+H] + = 200.2 @ 1.708 min (LC-MS method B).

[0024] A solution of 0002A (100 mg, 0.5 mmol) and iodomethane (355 mg, 2.5 mmol) in acetonitrile (5 mL) was stirred at 25°C for 16 hours. The reaction solution was concentrated to give 0002B (87 mg, 99% yellow solid). LCMS (ESI) [M+H] + =174 (LC-MS method A).

[0025] To a solution of 0002B (86 mg, 0.5 mmol) and 0006G (109 mg, 0.5 mmol) in dimethyl sulfoxide (3 mL) was added cesium carbonate (488 mg, 1.5 mmol) and heated to 85°C for 16 hours. The reaction solution was extracted with ethyl acetate (10 mL × 2), and the organic phase was washed with water (10 mL × 3) and saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was separated and purified by flash chromatography (ethyl acetate / petroleum ether = 0-50%) to obtain 0002C (40 mg, 22% as a yellow solid). LCMS (ESI) [M+H] + =356.1@1.296 min (LC-MS method A).

[0026] Potassium carbonate (69 mg, 0.5 mmol) was added to a solution of 0002C (35 mg, 0.1 mmol) in dimethyl sulfoxide (1 mL). The reaction mixture was cooled to 10°C and stirred for 5 minutes. Hydrogen peroxide (0.5 mL, 30%) was slowly added and allowed to react at room temperature for 10 minutes. The reaction was quenched with water and extracted with ethyl acetate (10 mL x 2). The organic phase was washed with water (10 mL x 3) and saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. 0002D (37 mg, 99%) was obtained and used directly in the next step without purification. LCMS (ESI) [M+H] +=374.1@1.69 min (LC-MS method B).

[0027] A solution of 0002D (37 mg, 0.1 mmol) and cyanamide (42 mg, 10 mmol) in ethanol (2 mL) was reacted at room temperature for 16 hours, concentrated, and the residue was purified by flash chromatography (methanol / dichloromethane = 0-8%) to give SB-0002 (30 mg, 82% as a white solid). LCMS (ESI) [M+H] + =368.2@5.716min (Method D). 1 HNMR(400MHz,DMSO-d6)δ9.623(bs,1H),8.182(s,1H),7.922(d,J=5.6Hz,1H),7.855(bs,1H),7.720(bs,1H),7.454-7.467(m,2H),4.432-4. 570(m,2H),4.246(s,1H),3.990(s,3H),2.990-3.023(m,1H),2.401-2 .680(m,2H),1.574-1.639(m,1H),1.365-1.432(m,1H),0.938(t,3H).

[0028] Example 2

[0029]

[0030] A solution of (S)-5-(Hydroxymethyl)pyrrolidin-2-one (3.45 g, 30 mmol), 2,2-dimethoxypropane (6.24 g, 60 mmol), and p-toluenesulfonic acid (190 mg, 1 mmol) in toluene (50 mL) was heated under reflux for 16 hours and then cooled to room temperature. The reaction solution was concentrated, and the residue was purified by flash chromatography (methanol / dichloromethane = 0-20%) to obtain 0006A (4.4 g, 94%, yellow oil). LCMS (ESI) [M+H] + =156.2@1.17min (Method A).

[0031] A solution of 0006A (1.55 g, 10 mmol) and Lawesson's reagent (2.02 g, 5 mmol) in toluene (20 mL) was heated under reflux for 16 hours and then cooled to room temperature. The reaction solution was concentrated, and the residue was purified by flash chromatography (ethyl acetate / petroleum ether = 0-50%) to obtain 0006B (250 mg, 15%, as a white solid). LCMS (ESI) [M+H] + =172.2@1.38min (Method B).

[0032] A solution of 0006B (171 mg, 1 mmol) and iodomethane (1.42 g, 10 mmol) in acetonitrile (10 mL) was reacted at 25°C for 16 hours. The reaction solution was concentrated to give 0006C (145 mg, 99%, white solid). LCMS (ESI) [M+H] + =146.1@0.97min (Method B).

[0033] To a solution of 4-bromo-3-methoxybenzaldehyde (6.45 g, 30 mmol) in toluene (150 mL) was added 2,2-dimethoxyethane-1-amine (4.7 g, 45 mmol). The reaction mixture was heated under reflux for 4 hours while separating the generated water with a water separator. The mixture was cooled to 0°C, maintaining the internal temperature below 5°C, and trifluoroacetic anhydride (12 mL, 90 mmol) and boron trifluoride etherate (11.1 mL, 90 mmol) were added sequentially. The mixture was warmed to room temperature and the reaction continued for 16 hours. The reaction mixture was poured into a mixture of ice and aqueous ammonia and extracted with ethyl acetate (200 mL x 3). The organic phase was washed sequentially with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography (ethyl acetate / petroleum ether = 1 / 1) to yield 0006D (4.7 g, 66%, dark brown solid). LCMS (ESI) [M+H] + =238.1@1.71min (Method B).

[0034] A solution of 0006D (1.5 g, 6.3 mmol), zinc powder (41 mg, 0.63 mmol), zinc cyanide (1.158 g, 9.9 mmol), 1,1'-bis(diphenylphosphino)ferrocene (35 mg, 0.063 mmol), and (dibenzylacetone)dipalladium(0) (29 mg, 0.315 mmol) in N,N-dimethylacetamide (12 mL) was heated to 150°C and microwaved for 1.5 hours. The reaction solution was filtered, and water was added to the filtrate. Solid precipitated, which was filtered and the filter cake dried to give 0006E (1.1 g, 94%, yellow solid). LCMS (ESI) [M+H] + =185.1@1.40min (Method B).

[0035] To a solution of 0006E (1.1 g, 5.98 mmol) in dichloromethane (20 mL) was added 3-chloroperoxybenzoic acid (1.547 g, 8.97 mmol) and the mixture was allowed to react at room temperature for 16 hours. Water was added to the reaction solution, which was extracted with dichloromethane (20 mL x 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to afford 0006F (1.2 g, 99%, yellow solid), which was used directly in the next step without purification. LCMS (ESI) [M+H] + =201.1@1.05min (Method B).

[0036] A mixture of 0006F (1.2 g, 6 mmol) and phosphorus oxychloride (10 mL) was heated to 105°C for 2 hours. The reaction solution was concentrated to dryness, and the residue was washed with saturated sodium bicarbonate solution (10 mL x 2) and dried to give 0006G (850 mg, 64%, light yellow solid). LCMS (ESI) [M+H] + =219.0@1.86min (Method A).

[0037] To a solution of 0006G (22 mg, 0.1 mmol) in dimethyl sulfoxide (1 mL) were added potassium carbonate (69 mg, 0.5 mmol) and hydrogen peroxide (0.2 mL, 30%), and the mixture was allowed to react at room temperature for 30 minutes. The reaction was quenched with water, and the reaction solution was extracted with ethyl acetate (10 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to afford 0006H (24 mg, 99%, yellow solid), which was used directly in the next reaction without purification. LCMS (ESI) [M+H] + =237.0@1.38min.(Method A).

[0038] Potassium bis(trimethylsilyl)amide (0.5 M / toluene, 1 mL, 0.5 mmol) was added to a solution of 0006H (24 mg, 0.1 mmol) and 0006C (15 mg, 0.1 mmol) in dry N,N-dimethylformamide (2 mL) at -10°C. The reaction mixture was allowed to warm to room temperature and stirred for 4 hours. The reaction mixture was extracted with ethyl acetate (10 mL × 2). The organic phase was washed with water (10 mL × 3) and saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was separated and purified by flash chromatography (methanol / dichloromethane = 0-10%) to obtain 0006I (20 mg, 57%, yellow solid). LCMS (ESI) [M+H] + =346.2@1.47min (Method B).

[0039] A solution of 0006I (20 mg, 0.058 mmol) and cyanamide (24 mg, 0.58 mmol) in ethanol (3 mL) was reacted at room temperature for 16 hours. The reaction solution was concentrated, and the residue was purified by flash chromatography (methanol / dichloromethane = 0-7%) to obtain a crude product. This was further purified by Pre-HPLC (Column Xbridge 21.2*250mm C18, 10 μm, mobile phase A: water (10 mmol / L ammonium bicarbonate) B: acetonitrile). SB-0006 (3 mg, 15%, white solid) was obtained. LCMS (ESI) [M+H] + =340.2@4.71min (Method D). 1HNMR(400MHz,CD3OD)δ8.421(s,1H),7.921(d,J=5.6Hz,1H),7.690(s,1H),7.388(d,J=5.6Hz,1H),4.554-4.6 41(m,2H),4.444-4.474(m,1H),3.119(s,3H),2.932-3.151(m,2H),2.508-2.559(m,1H),2.217-2.232(m,1H).

[0040] Example 3

[0041]

[0042] A solution of (3S,4S,5S)-4-ethyl-3-fluoro-5-(hydroxymethyl)pyrrolidin-2-one (160 mg, 1 mmol), 2,2-dimethoxypropane (208 mg, 2 mmol), and p-toluenesulfonic acid (6 mg, 0.03 mmol) in toluene (50 mL) was heated under reflux for 16 hours and then cooled to room temperature. The reaction solution was concentrated, and the residue was purified by flash chromatography (methanol / dichloromethane = 0-20%) to obtain 0011A (160 mg, 80%, yellow oil). LCMS (ESI) [M+H] + =202.2@1.31min (Method B).

[0043] A solution of 0011A (0.2 g, 1 mmol) and Lawesson's reagent (202 mg, 0.5 mmol) in toluene (10 mL) was heated under reflux for 4 hours and then cooled to room temperature. The reaction solution was concentrated, and the residue was purified by flash chromatography (ethyl acetate / petroleum ether = 0-50%) to obtain 0011B (120 mg, 55%, yellow oil). LCMS (ESI) [M+H] + =218.1@1.83min (Method A).

[0044] A solution of 0011B (120 mg, 0.55 mmol) and iodomethane (780 mg, 5.5 mmol) in acetonitrile (5 mL) was reacted at 25°C for 16 hours. The reaction solution was concentrated to give 0011C (105 mg, 99%, white solid). LCMS (ESI) [M+H] + =192.1@1.34min (Method B).

[0045] Potassium bis(trimethylsilyl)amide (0.5M / toluene, 2.1 mL, 1.05 mmol) was added to a solution of 0006H (62 mg, 0.26 mmol) and 0011C (50 mg, 0.26 mmol) in dry N,N-dimethylformamide (2 mL) at -10°C. The reaction mixture was heated to 25°C for 4 hours. The mixture was extracted with ethyl acetate (10 mL × 3), and the organic phase was washed with water (10 mL × 3) and saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography (methanol / dichloromethane = 0-6%) to give 0011D (100 mg, 97%, yellow solid). LCMS (ESI) [M+H] + =392.1@1.68min (Method B).

[0046] A solution of 0011D (100 mg, 0.255 mmol) and cyanamide (107 mg, 2.55 mmol) in ethanol (5 mL) was reacted at room temperature for 16 hours. The reaction solution was concentrated, and the residue was purified by preparative Pre-HPLC (Column Xbridge 21.2*250 mm C18, 10 μm, mobile phase A: water (10 mmol / L ammonium bicarbonate) B: acetonitrile). SB-0011 (2 mg, 2%, white solid) was obtained. LCMS (ESI) [M+H] + =386.2@6.15min (Method D). 1 HNMR(400MHz,CD3OD)δ8.375(s,1H),7.941(d,J=5.6Hz,1H),7.498(s,1H),7.421(d,J=5.6Hz,1H),4.776-4.8 52(m,1H),4.391-4.499(m,2H),4.135(s,3H),2.787-2.862(m,2H),1.700-1.772(m,2H),1.109-1.145(m,3H).

[0047] Attached analysis method

[0048] 1 H NMR and 19 F NMR spectra were detected on a Bruker AVIII400.

[0049] LCMS was performed on an Agilent 1200HPLC / 6100SQ system using the following conditions:

[0050] Method A: Mobile phase: A: water (0.01% TFA) B: acetonitrile (0.01% TFA); Gradient phase: 5% B increase to 95% B within 1.4 min, 95% B with 1.6 min (total run time: 3 min); Flow rate: 2.3 mL / min; Column type: SunFire C18, 4.6*50 mm, 3.5 μm; Column temperature: 50°C. Detectors: ADC ELSD, DAD (214 nm and 254 nm), ES-API.

[0051] Method B: Mobile phase: A: water (10 mM NH4HCO3) B: acetonitrile; Gradient phase: 10% to 95% B within 1.5 min, 95% B with 1.5 min (total run time: 3 min); Flow rate: 1.8 mL / min; Column type: XBridgeC18, 4.6*50 mm, 3.5 um; Column temperature: 50°C. Detectors: ADC ELSD, DAD (214 nm and 254 nm), MSD (ES-API).

[0052] Method C: Mobile phase: A: water (0.01% TFA) B: acetonitrile (0.01% TFA); Gradient phase: 5% B increase to 95% B within 1.2 min, 95% B with 13.8 min (total run time: 15 min); Flow rate: 2 mL / min; Column type: SunFire C18, 4.6*50 mm, 3.5 μm; Column temperature: 50°C. Detectors: UV (214 nm and 4 nm), MS (ESI, Posmode, 132 to 1500 amu).

[0053] Method D: Mobile phase: A: water (10 mM NH4HCO3) B: acetonitrile; Gradient phase: 10% to 95% B within 8 min, 95% B with 7 min (total run time: 15 min); Flow rate: 1 mL / min; Column type: XBridge C18, 4.6*50 mm, 3.5 um; Column temperature: 40°C. Detectors: UV (214 nm and 4 nm), MS (ESI, Pos mode, 132 to 1500 amu).

[0054] Method E: Mobile phase: A: water (0.01% TFA) B: acetonitrile (0.01% TFA); Gradient phase: 5% B increase to 95% B within 1.2 min, 95% B for 1.8 min, back to 5% B within 0.01 min. (Total run time: 15 min); Flow rate: 2 mL / min; Column type: SunFire C18, 4.6*50 mm, 3.5 μm; Column temperature: 40°C. Detectors: UV (214 nm and 4 nm), MS (ESI, Pos mode, 132 to 1500 amu).

[0055] Effect Example IRAK4 kinase activity inhibition detection

[0056] Detection of IC of IRAK4 kinase 50 Staurosporine was used as a positive control compound. This experiment used a mobility shift assay to screen compounds on IRAK4 kinase.

[0057] Preparation of a compound concentration gradient: Test compound starting at 10 μM, diluted 4-fold, 10 times the final concentration, and tested in duplicate. Serially dilute the test compound in a 384-well plate to 10 different concentrations, each 100 times the final concentration. Then, using a pipetting system (Labcyte, Echo550), transfer 250 nL of the solution to the 384-well plate for later use. Add 250 nL of 100% DMSO to each of the negative and positive control wells. Prepare a kinase (Carna, 09-145) solution at 2.5 times the final concentration using kinase buffer (prepared by Sundia Pharmaceuticals (Shanghai) Co., Ltd.). Add 10 μL of the kinase solution at 2.5 times the final concentration to each of the compound wells and the positive control wells (Staurosporine, selleckchem, S1421); add 10 μL of kinase buffer to the negative control wells. Centrifuge at 1000 rpm for 30 seconds, shake to mix, and incubate at room temperature for 10 minutes. Prepare a mixture of ATP (Sigma, A2383) and substrate (Kinase substrate 8, GL, 112396) at a final concentration of 25 / 15 times using kinase buffer. Add 15 μL of this mixture of ATP and substrate at a final concentration of 25 / 15 times to initiate the reaction. Centrifuge the 384-well plate at 1000 rpm for 30 seconds, vortex to mix, and incubate at room temperature for the appropriate time. Add 30 μL of stop detection solution (prepared by Sundia Pharmaceutical Technology (Shanghai) Co., Ltd.) to stop the kinase reaction, centrifuge at 1000 rpm for 30 seconds, and vortex to mix. Read the conversion rate using a microplate reader (Caliper EZ Reader II).

[0058] Data Analysis:

[0059] Calculation formula:

[0060]

[0061] %Inhibition is the inhibition rate; Conversion%_sample is the conversion rate reading of the sample;

[0062] Conversion%_min: Mean value of negative control wells, representing the conversion rate reading of wells without enzyme activity;

[0063] Conversion%_max: The mean of the positive control wells, representing the conversion rate readings of wells without compound inhibition.

[0064] Fitting the dose-effect curve:

[0065] The logarithmic concentration value was plotted on the X-axis, and the percentage inhibition rate was plotted on the Y-axis. The log(inhibitor) vs. response-variable slope function of the GraphPad Prism 5 analysis software was used to fit the dose-effect curve to obtain the IC50 value of each compound on the enzyme activity. The calculation formula is as follows: Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC 50 -X)*HillSlope)), the results are shown in Table 1:

[0066] Table 1

[0067] Compound number <![CDATA[IC 50 (nM)]]> SB-0002 0.84 SB-0006 1.77 SB-0011 0.94

Claims

1. A compound as described below or a pharmaceutically acceptable salt thereof:

2. A pharmaceutical composition comprising the compound as claimed in claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

3. Use of the compound according to any one of claims 1 to 2 or a pharmaceutically acceptable salt thereof, and the pharmaceutical composition thereof, in the preparation of IRAK4 kinase inhibitor drugs.

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