Deuterated indole urea compound and medical use thereof

Abstract

A deuterated indole urea compound as shown in formula I and the medical use thereof as a STING inhibitor. The compound of formula I or a pharmaceutically acceptable salt thereof can be used in the preparation of a drug for preventing or treating STING-mediated diseases.

Description

Deuterated indole urea compounds and their medical uses Technical Field

[0001] The present invention belongs to the field of biomedicine, and specifically relates to a class of deuterated indole urea compounds and their preparation methods, medical uses, and pharmaceutical compositions as novel STING inhibitors. Background Art

[0002] H-151, also known as 1-(4-ethylphenyl)-3-(1H-indol-3-yl)urea (having the following structure), is an indole STING palmitoylation inhibitor (Nature 2018, 559, 269-273; EP3556362), which can significantly improve the symptoms of ALS (Cell 2020, 183, 636-649). The applicant's research has shown that H-151 has significant therapeutic effects in psoriasis models (Bri J Pharmacol, 2021, 178, 4907-4922). However, H-151 has poor metabolic stability, resulting in poor pharmacokinetic properties, which severely limits its clinical application. So far, no STING inhibitor has entered the clinical research stage, and there is a very urgent clinical need. In short, this field still needs to develop STING inhibitors with strong activity and metabolic stability.

[0003] Summary of the Invention

[0004] Purpose of the Invention: To address the challenges of the prior art, the present invention provides a novel class of deuterated indole urea compounds with excellent STING inhibitory activity and pharmacokinetic properties, which can be used to prepare therapeutics for the prevention or treatment of STING-mediated diseases. The novel deuterated indole urea compounds provided by the present invention not only significantly improve the metabolic stability of STING inhibitor compounds such as H-151, but also further enhance their inhibitory activity.

[0005] The present invention also provides a preparation method and application of the deuterated indole urea compound and a pharmaceutical composition.

[0006] Technical solution: In order to achieve the above-mentioned object, the present invention provides a deuterated indole urea compound or a pharmaceutically acceptable salt, solvate or prodrug thereof as shown in the following formula I:

[0007] Among them, Y 1 ,Y 2 ,Y 3 ,Y 4 ,Y 5 ,Y 6 ,Y 7 ,Y 8 ,Y 9 ,Y10 ,Y 11 ,Y 12 ,Y 13 or Y 14 are independently selected from hydrogen or deuterium; and Y 1 ,Y 2 ,Y 3 ,Y 4 ,Y 5 ,Y 6 ,Y 7 ,Y 8 ,Y 9 ,Y 10 ,Y 11 ,Y 12 ,Y 13 or Y 14 At least one of them is deuterium.

[0008] Preferably, the Y 1 , Y 2 , Y 3 , Y 4 , Y 5 , Y 6 , Y 7 , Y 8 , Y 9 , Y 10 , Y 11 , Y 12 , Y 13 or Y 14 are independently selected from hydrogen or deuterium; and Y 6 , Y 7 , Y 8 , Y 9 , Y 10 , Y 11 , Y 12 , Y 13 or Y 14 At least one of them is deuterium.

[0009] Furthermore, the Y 1 , Y 2 , Y 3 , Y 4 , Y 5 , Y 6 , Y 7 , Y 8 , Y 9 , Y 10 , Y 11 , Y 12 , Y 13 or Y 14 ; and Y 10 , Y 11 , Y 12 , Y 13 or Y14 At least one of them is deuterium.

[0010] Wherein, the compound or its pharmaceutically acceptable salt is selected from any one of the following Table 1:

[0011] Table 1. Structure and nomenclature of deuterated indole urea compounds

[0012] The compounds of the present invention can be used as pharmaceutically acceptable salts. The salt can be an acid salt of at least one of the following acids: galactaric acid, D-glucuronic acid, glycerophosphoric acid, hippuric acid, isethionic acid, lactobionic acid, maleic acid, 1,5-naphthalenedisulfonic acid, naphthalene-2-sulfonic acid, pivalic acid, terephthalic acid, thiocyanic acid, bile acid, n-dodecylsulfuric acid, benzenesulfonic acid, citric acid, D-glucose, glycolic acid, lactic acid, malic acid, malonic acid, mandelic acid, phosphoric acid, propionic acid, hydrochloric acid, sulfuric acid, tartaric acid, succinic acid, formic acid, hydroiodic acid, hydrochloric acid Bromic acid, methanesulfonic acid, nicotinic acid, nitric acid, orotic acid, oxalic acid, picric acid, L-pyroglutamic acid, saccharinic acid, salicylic acid, gentisic acid, p-toluenesulfonic acid, valeric acid, palmitic acid, sebacic acid, stearic acid, lauric acid, acetic acid, adipic acid, carbonic acid, benzenesulfonic acid, ethanedisulfonic acid, ethylsuccinic acid, fumaric acid, 3-hydroxynaphthalene-2-carboxylic acid, 1-hydroxynaphthalene-2-carboxylic acid, oleic acid, undecylenic acid, ascorbic acid, camphoric acid, camphorsulfonic acid, dichloroacetic acid, ethanesulfonic acid. On the other hand, the salt can also be a salt formed by a compound of the present invention and a metal (including sodium, potassium, calcium etc.) ion or a pharmaceutically acceptable amine (including ethylenediamine, tromethamine etc.), ammonium ion or choline.

[0013] The compounds of the present invention or pharmaceutically acceptable salts thereof can be prepared by referring to the methods described in the examples or by modified methods.

[0014] The present invention provides use of any one of the compounds shown in Formula I and Table 1, or a pharmaceutically acceptable salt or prodrug thereof, as a STING inhibitor.

[0015] The present invention provides use of any one of the compounds shown in Formula I and Table 1, or a pharmaceutically acceptable salt or prodrug thereof, in the preparation of a medicament for preventing or treating STING-mediated diseases.

[0016] Among them, the STING-mediated diseases include infectious diseases, inflammatory diseases, autoimmune diseases, metabolic diseases, organ fibrosis diseases, cardiovascular and cerebrovascular diseases, respiratory diseases, central nervous system diseases, cancer or precancerous syndromes.

[0017] Among them, the infectious diseases include but are not limited to Mycobacterium tuberculosis infection, Chlamydia infection, herpes virus (herpes simplex virus) infection, adenovirus infection, hepatitis B virus infection, orthomyxovirus infection and coronavirus infection.

[0018] Among them, the inflammatory diseases include but are not limited to musculoskeletal muscle inflammation (inflammation of the hands, wrists, elbows, shoulders, neck, knees, ankles and feet, such as osteoarthritis, rheumatoid arthritis, ankylosing spondylitis, acute and chronic infectious arthritis, etc.), eye inflammation (keratitis, scleritis, conjunctivitis, etc.), digestive system inflammation (colitis, hepatitis, primary biliary cholangitis, primary sclerosing cholangitis, cholecystitis, pancreatitis, gastritis, enteritis, inflammatory bowel disease, proctitis), nervous system inflammation (meningitis, neuromyotonia, polymyositis, etc.), sclerosis, CNS vasculitis), inflammation of the vascular system or lymphatic system (vasculitis, lymphangitis, phlebitis), reproductive system inflammation (cervicitis, endometritis, epididymitis, orchitis, urethritis), respiratory system inflammation (pneumonia, asthma, chronic obstructive pulmonary disease, chronic bronchitis, emphysema, bronchiolitis obliterans, idiopathic pulmonary fibrosis, cystic fibrosis), other inflammatory diseases including appendicitis, myocarditis, mumps, gingivitis, prostatitis, peritonitis, pleurisy, vasculitis, phlebitis, dermatitis or edema, etc.

[0019] Among them, the autoimmune diseases include but are not limited to ulcerative colitis, Crohn's disease, systemic lupus erythematosus, familial pernio lupus, lupus nephritis, Chagas disease, rheumatoid arthritis, psoriasis, vitiligo, alopecia areata, multiple sclerosis, scleroderma, Behcet's disease, STING-associated vasculitis (SAVI) of infancy, Aicardi-Goutières syndrome or retinal vasculopathy with cerebral protein dystrophy (RCVL), etc.

[0020] Among them, the metabolic diseases include but are not limited to non-alcoholic fatty liver disease, alcoholic fatty liver disease, insulin resistance, metabolic syndrome, diabetes and its complications, polycystic kidney disease, polycystic ovary syndrome, hyperlipidemia, obesity, hyperuricemia, gout or osteoporosis, etc.

[0021] Among them, the respiratory system diseases include but are not limited to cough, asthma, tracheitis, bronchitis, pneumonia, respiratory distress syndrome, emphysema, chronic obstructive pulmonary disease, idiopathic pulmonary fibrosis, pulmonary hypertension, cystic fibrosis or rhinitis, etc.

[0022] The organ fibrosis diseases include, but are not limited to, liver fibrosis, liver cirrhosis, pulmonary fibrosis or renal fibrosis.

[0023] Among them, the cardiovascular and cerebrovascular diseases include but are not limited to hypertension, atherosclerosis, peripheral vascular disease, coronary heart disease, angina pectoris, ischemia, cardiac ischemia, stroke, myocardial infarction, cardiomyopathy, heart failure, injurious reperfusion, restenosis after angioplasty, ischemic encephalopathy, stroke, hemorrhagic encephalopathy, cerebral hemorrhage, cerebral edema, or cerebral infarction, etc.

[0024] Among them, the central nervous system diseases include but are not limited to Parkinson's disease, Alzheimer's disease, α-synuclein disease, depression, amyotrophic lateral sclerosis (ALS), fibromyalgia syndrome, neuralgia, Down syndrome, Hallervorden-Spanish disease, Huntington's disease and Wilson's disease.

[0025] The cancer includes, but is not limited to, cancer of the lung, bone, pancreas, liver, kidney, head, uterus, ovary, stomach, colon, esophagus, small intestine, endocrine system, prostate, bladder, cervix, and vagina, such as liver cancer, kidney cancer, cervical cancer, lung cancer, skin cancer, uterine cancer, adenocarcinoma, prostate cancer, sarcoma, osteosarcoma, thyroid cancer, non-small cell lung cancer, esophageal cancer, chronic myeloid leukemia, chronic lymphocytic leukemia, acute myeloid leukemia, acute lymphocytic leukemia, multiple myeloma, malignant lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, and neuroblastoma.

[0026] Preferably, the autoimmune disease is psoriasis; and the inflammatory disease is dermatitis.

[0027] Any compound of Formula I and Table 1 of the present invention or a pharmaceutically acceptable salt or prodrug thereof can be used to prepare an immune adjuvant drug.

[0028] Any compound of Formula I and Table 1 of the present invention or a pharmaceutically acceptable salt or prodrug thereof can be used to prevent or treat T cell-mediated hypersensitivity reactions with inflammatory components, including urticaria, skin allergies, allergic rhinitis, contact dermatitis and respiratory allergies.

[0029] The compounds of the present invention can be used alone or in combination with other therapeutic agents. As immunomodulators, the compounds of the present invention can be used as monotherapy or in combination with other therapeutic agents to treat STING-mediated diseases.

[0030] The present invention provides a pharmaceutical composition for preventing or treating a STING-mediated disease, comprising a therapeutically effective amount of any compound of Formula I and Table 1, or a pharmaceutically acceptable salt or prodrug thereof, as an active ingredient and a pharmaceutically acceptable carrier. The carriers that can be mixed arbitrarily can vary depending on the dosage form, administration form, and the like. Examples of carriers include excipients, binders, disintegrants, lubricants, flavoring agents, fragrances, colorants, or sweeteners. The pharmaceutical composition can be in the form of a capsule, powder, tablet, granule, pill, injection, syrup, oral liquid, inhalant, ointment, solution, cream, gel, powder, lotion, tincture, suppository, or patch, among other conventional pharmaceutical formulations.

[0031] Deuterium is a safe, stable, and non-radioactive isotope of hydrogen. Compared to hydrogen, deuterium forms stronger chemical bonds with carbon. This invention utilizes a specific deuterated modification strategy to significantly improve the metabolic stability of the STING inhibitor compound H-151, demonstrating excellent therapeutic efficacy in related indications, such as psoriasis.

[0032] Beneficial effects: Compared with the prior art, the present invention has the following advantages:

[0033] The STING inhibitor H-151 reported in the literature (Nature 2018, 559, 269-273; EP3556362) has weak STING inhibitory activity (THP1-Dual IC 50 =914nM) and poor metabolic stability (a liver microsomal half-life of only 6.75 minutes), which seriously affects its drugability. The deuterated compounds of Formula I specifically designed in this invention exhibit potent STING inhibitory activity, with most compounds showing superior STING inhibitory activity to H-151. Importantly, studies on metabolic stability in human liver microsomes demonstrate that the compounds of this invention possess improved metabolic stability, thereby enhancing their drugability. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 shows the therapeutic effect of compound 15 on a mouse psoriasis model;

[0035] Figure 2 is a schematic diagram of the PASI (Psoriasis Area and Severity Index) scoring of compound 15 in the treatment of psoriasis mouse model (n=5, ***p<0.001 compared with the model group at the same time).

[0036] Figure 3 shows the therapeutic effects of compounds 6, 15, 21 and benvimod on a mouse psoriasis model;

[0037] Figure 4 is a schematic diagram of the PASI (Psoriasis Area and Severity Index) scores for the treatment of mouse psoriasis models with compounds 6, 15, 21 and benvimod (n=5, **p<0.01, ***p<0.001 for the benvimod group, compound 6 group, compound 15 group, and compound 21 group compared with the model group at the same time). DETAILED DESCRIPTION

[0038] The present invention is further illustrated by the following examples. The following examples are intended to better illustrate the present invention and are not intended to limit its scope. Various changes and modifications may be made to the present invention without departing from its spirit and scope.

[0039] The raw materials and equipment used in the specific embodiments of the present invention are all known products and are purchased commercially.

[0040] The structures of the compounds were determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). NMR measurements were performed using a Bruker NMR spectrometer in deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), or deuterated methanol (CD3OD) as the solvent, and tetramethylsilane (TMS) as the internal standard.

[0041] Silica gel column chromatography generally uses 200-300 mesh silica gel from Qingdao Ocean Chemical Plant Branch as the carrier.

[0042] The known starting materials of the present invention can be synthesized by methods known in the art, or can be purchased from companies such as Leyan, Bid Pharmaceuticals, Aladdin, and Anaiji.

[0043] Example 1

[0044] 1-(4-Ethylphenyl)-3-(1H-indol-3-yl-4,5,6-7-d4)urea

[0045] Synthesis of intermediate A1

[0046] A two-necked flask was placed under argon atmosphere and a solution of nitrobenzene-d5 (641 mg, 5 mmol) in anhydrous tetrahydrofuran (THF) (5 mL) was added. Vinylmagnesium bromide (1 M in THF, 16 mL) was added dropwise at -40°C and stirred for 2 h at -40°C. After completion of the reaction, saturated sodium chloride solution (50 mL) was added to the reaction mixture, which was maintained at -40°C to quench the reaction. The aqueous phase was extracted with ethyl acetate (50 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 30:1) to obtain Intermediate A1 (orange solid, 65 mg) in an 11% yield.

[0047] Synthesis of intermediate A2

[0048] Intermediate A1 (455 mg, 3 mmol) was dissolved in N,N-dimethylformamide (DMF) (9 mL). Trifluoroacetic anhydride (TFAA) (2.52 g, 12 mmol) was added dropwise in an ice bath and stirred at room temperature for 4 hours. After the reaction, water (30 mL) was added to quench the reaction mixture, resulting in the precipitation of a pink solid. The mixture was stirred at room temperature for 1 hour. Filtration afforded the crude intermediate A2, which was used directly in the next reaction without further purification.

[0049] Synthesis of intermediate A3

[0050] The crude intermediate A2 was dissolved in 20% NaOH (9 mL) and the reaction mixture was placed in an oil bath and allowed to react at 100°C for 4 hours. After the reaction, insoluble impurities were removed by filtration. The filtrate was adjusted to pH 4 with 6N HCl solution. A yellow solid precipitated and was filtered to obtain intermediate A3 (yellow solid, 500 mg). The two-step yield was 78%.

[0051] Synthesis of intermediate A4

[0052] Intermediate A3 (500 mg, 2.56 mmol) was dissolved in dichloromethane (DCM) (9 mL), and triethylamine (TEA) (709 μL, 5.1 mmol) was added. The mixture was stirred at room temperature for 15 minutes, and diphenylphosphoryl azide (DPPA) (633 mg, 2.3 mmol) was slowly added. The reaction was allowed to react overnight. After completion of the reaction, 1N HCl solution (6 mL) was added, and the mixture was extracted with dichloromethane (20 mL × 3). The organic phases were combined, washed with saturated brine (30 mL × 1), dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 2:1) to obtain Intermediate A4 (pink solid, 330 mg) in a 71% yield.

[0053] Synthesis of compound 1

[0054] Intermediate A4 (50 mg, 0.23 mmol) was dissolved in toluene (PhMe) (3 mL). The mixture was placed in an oil bath and allowed to react at 100°C for 3 hours. The mixture was then cooled to room temperature and 4-ethylaniline (38 mg, 0.20 mmol) was added. The reaction was allowed to proceed overnight, resulting in the precipitation of a white solid. The solvent was then evaporated under reduced pressure. Petroleum ether (5 mL) and ethyl acetate (1 mL) were added to the residue, stirred at room temperature for 2 hours, and filtered to afford compound 1 (white solid, 36 mg) in a 65% yield. 1H NMR(400MHz,DMSO-d6)δ9.87(s,1H),7.65(s,1H),7.57(s,1H),6.66(d,J=2.4Hz,1H),6.59-6.5 1(m,2H),6.32-6.23(m,2H),1.72(t,J=7.6Hz,2H),0.33(t,J=7.6Hz,3H).HRMS(ESI)calcd.for C 17 H 13 D4N3O[M+H] + 284.1701, found 284.1715.

[0055] Example 2

[0056] 1-(4-ethylphenyl-2,3,5,6-d4)-3-(1H-indol-3-yl)urea

[0057] Synthesis of intermediate B1

[0058] In an ice bath, 98% concentrated sulfuric acid (1.9 mL) was added dropwise to 65% concentrated nitric acid (1.9 mL) to prepare a mixed acid. Bromobenzene-d5 (2.2 g, 13.5 mmol) was then added dropwise to the mixed acid in an ice bath. The system was then transferred to an oil bath and reacted at 45°C for 15 minutes. After the reaction, the mixture was cooled to 0°C and ice water (6 mL) was added. A white solid was produced, which was extracted with ethyl acetate (10 mL x 3). The organic phase was concentrated and recrystallized from anhydrous ethanol to obtain Intermediate B1 (white solid, 776 mg) in a 31% yield.

[0059] Synthesis of intermediate B2

[0060] Potassium vinyl trifluoroborate (662 mg, 4.94 mmol), cesium carbonate (3.2 g, 9.87 mmol), triphenylphosphine (52 mg, 0.20 mmol), and palladium chloride (12.4 mg, 0.07 mmol) were weighed into a three-necked flask. The atmosphere was purged with argon three times. A mixture of anhydrous tetrahydrofuran (THF) solution (10 mL) and water (1 mL) containing B1 (679 mg, 3.29 mmol) was added. The mixture was transferred to an oil bath and reacted at 70°C for 24 hours. After completion of the reaction, the solvent was evaporated under reduced pressure, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 50:1) to obtain intermediate B2 (a yellow oily liquid, 450 mg) in a 67% yield.

[0061] Synthesis of intermediate B3

[0062] Compound B2 (791 mg, 3.06 mmol) was dissolved in ethanol (15 mL) and 10% palladium on carbon (80 mg) was added. The atmosphere was replaced with hydrogen three times, and the reaction mixture was allowed to react at room temperature overnight. After the reaction was completed, the filtrate was filtered and the solvent was evaporated under reduced pressure to obtain a crude product containing compound B3, which was used directly in the next reaction without further purification.

[0063] Synthesis of intermediate B4

[0064] 3-Indolecarboxylic acid (500 mg, 2.56 mmol) was dissolved in dichloromethane (DCM) (9 mL), and triethylamine (TEA) (709 μL, 5.1 mmol) was added. The mixture was stirred at room temperature for 15 minutes, and diphenylphosphoryl azide (DPPA) (633 mg, 2.3 mmol) was slowly added. The reaction was allowed to react overnight. After completion of the reaction, 1N HCl solution (6 mL) was added, and the mixture was extracted with dichloromethane (20 mL × 3). The organic phases were combined, washed with saturated brine (30 mL × 1), dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 2:1) to obtain Intermediate B4 (pink solid, 330 mg) with a two-step yield of 75%.

[0065] Synthesis of compound 2

[0066] Intermediate B4 (50 mg, 0.23 mmol) was dissolved in toluene (PhMe) (3 mL). The mixture was placed in an oil bath and allowed to react at 100°C for 3 hours. The mixture was then cooled to room temperature and B3 (38 mg, 0.20 mmol) was added. The reaction was allowed to proceed overnight, resulting in the precipitation of a white solid. The solvent was then evaporated under reduced pressure. Petroleum ether (5 mL) and ethyl acetate (1.5 mL) were added to the residue, stirred at room temperature for 2 hours, and filtered to obtain compound 2 (white solid, 36 mg) in a 65% yield. 1 H NMR (300MHz, DMSO-d6) δ10.72(s,1H),8.49(s,1H),8.41(s,1H),7.57-7.46(m,2H),7.33(d,J=8.1Hz,1H ),7.13-7.06(m,1H),7.05-6.97(m,1H),2.60-2.51(m,2H),1.16(t,J=7.6Hz,3H).HRMS(ESI)calcd.for C 17 H 13 D4N3O[M+H] + 284.1701,found 284.1700.

[0067] Example 3

[0068] 1-(4-(ethyl-d5)phenyl-2,3,5,6-d4)-3-(1H-indol-3-yl)urea

[0069] Synthesis of intermediate C1

[0070] In an ice bath, 98% concentrated sulfuric acid (1.02 mL) was added dropwise to 65% concentrated nitric acid (0.85 mL) to prepare a mixed acid. In an ice bath, the mixed acid was slowly added dropwise to ethylbenzene-d 10 The mixture was stirred for 1 hour at room temperature. After the reaction, water (50 mL) was added for dilution and the mixture was extracted with ethyl acetate (40 mL x 2). The organic phases were combined, washed with saturated brine (30 mL x 1), dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether) to obtain intermediate C1 (colorless oily liquid, 114.5 mg) in a 31% yield.

[0071] Synthesis of intermediate C2

[0072] Compound C1 (114.5 mg, 0.7 mmol) was weighed and dissolved in ethanol (2 mL). 10% palladium on carbon (30 mg) was added, and the atmosphere was replaced with hydrogen three times. The system was allowed to react at room temperature overnight. After the reaction, the mixture was filtered, and the solvent was evaporated under reduced pressure to obtain a crude product containing compound C2, which was used directly in the next reaction without further purification.

[0073] Synthesis of compound 3

[0074] Referring to the synthesis method of Example 2, B3 in Example 2 was replaced by C2 to obtain compound 3 (white powder, 65 mg) with a yield of 73%: 1 H NMR(300MHz,DMSO-d6)δ10.72(s,1H),8.49(s,1H),8.41(s,1H),7.54-7.47(m,3H) ,7.33(d,J=8.1Hz,1H),7.12-7.06(m,1H),7.04-6.98(m,1H).HRMS(ESI)calcd.for C 17 H8D9N3O[M+H] + 289.2015,foun289.2015.

[0075] Example 4

[0076] 1-(4-(ethyl-d5)phenyl-2,3,5,6-d4)-3-(1H-indol-3-yl-4,5,6-7-d4)urea

[0077] Referring to the synthesis method of Example 1, 4-ethylaniline in Example 1 was replaced with C2 to obtain compound 4 (white powder, 50 mg) with a yield of 57%: 1 H NMR(300MHz,DMSO-d6)δ10.71(s,1H),8.49(s,1H),8.42(s,1H),7.49(d,J=2.4Hz,1H).HRMS(ESI)calcd.for C 17 H4D 13 N3O[M+H] + 293.2266,found 293.2262.

[0078] Example 5

[0079] 1-(4-ethylphenyl-2,3,5,6-d4)-3-(1H-indol-3-yl-4,5,6-7-d4)urea

[0080] Referring to the synthesis method of Example 1, the 4-ethylaniline in Example 1 was replaced with B3 to obtain compound 5 (white powder, 56 mg) with a yield of 65%: 1 H NMR(300MHz, DMSO-d6)δ10.71(s,1H),8.49(s,1H),8.42(s,1H),7.49(d,J=2.4Hz,1H),2.57-2.52(m,2H),1.14(d,J=7.6Hz,3H).HRMS(ESI)calcd.for C 17 H 10 D8N3O[M+H] + 288.1952,found288.1950.

[0081] Example 6

[0082] 1-(4-(ethyl-d5)phenyl)-3-(1H-indol-3-yl)urea

[0083] Synthesis of intermediate D1

[0084] Dissolve triphenylphosphine (2.7 g, 10 mmol) in tetrahydrofuran (THF) (20 mL) and slowly add deuterated iodomethane (1.6 g, 10.5 mmol) dropwise. A white solid precipitates. Stir at room temperature for 10 hours. After the reaction is complete, filter and obtain Intermediate D1 (white solid, 4.3 g) in an 89% yield.

[0085] Synthesis of intermediate D2

[0086] Weigh p-nitrobenzoic acid (1 g, 6 mmol) and sodium borodeuteride (0.5 g, 12 mmol) into a reaction flask and add tetrahydrofuran (15 mL). Slowly add boron trifluoride etherate (1.5 mL) dropwise under an ice bath. The mixture is slowly warmed to room temperature and allowed to react for 2 hours. After completion, quench the reaction by slowly adding water (40 mL) under an ice bath. Extract with ethyl acetate (40 mL x 2). The combined organic phases are washed with saturated brine (30 mL x 1), dried over anhydrous sodium sulfate, and the solvent removed under reduced pressure. This yields a crude product containing compound D2, which is used directly in the next reaction without further purification.

[0087] Synthesis of intermediate D3

[0088] D2 (930 mg, 6 mmol) and 2-iodobenzoic acid (IBX) (5.04 g, 18 mmol) were weighed and placed in a reaction flask. Ethyl acetate (50 mL) was added and the system was moved to an oil bath. The reaction was allowed to proceed at 80°C for 3 hours, followed by cooling to room temperature. After completion of the reaction, the filtrate was filtered, and the solvent was evaporated under reduced pressure. The filtrate was diluted with water (50 mL) and extracted with ethyl acetate (40 mL x 2). The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 15:1) to obtain intermediate D3 (white solid, 789 mg) with a two-step yield of 86.4%.

[0089] Synthesis of intermediate D4

[0090] D3 (651.6 mg, 1.60 mmol) and potassium carbonate (367.6 mg, 2.66 mmol) were weighed and placed in a Shrek tube. The atmosphere was purged with argon three times, and a solution of D3 (202.2 mg, 1.33 mmol) in anhydrous tetrahydrofuran (THF) (5 mL) was added. The system was moved to an oil bath and reacted at 70°C for 24 hours. After completion of the reaction, the filtrate was filtered and the solvent was evaporated under reduced pressure. The filtrate was diluted with water (50 mL) and extracted with ethyl acetate (40 mL × 3). The organic phases were combined, washed with saturated brine (30 mL × 1), dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 50:1) to obtain intermediate D4 (pale yellow oily liquid, 168.6 mg) in a yield of 78.7%.

[0091] Synthesis of intermediate D5

[0092] Compound D4 (131 mg, 1.1 mmol) was weighed and dissolved in ethanol (2 mL). 10% palladium on carbon (35 mg) was added, and the mixture was deuterated three times. The reaction mixture was allowed to react at room temperature overnight. After completion of the reaction, the mixture was filtered, and the solvent was evaporated under reduced pressure to obtain a crude product containing compound D5, which was used directly in the next reaction without further purification.

[0093] Synthesis of compound 6

[0094] Referring to the synthesis method of Example 1, the 4-ethylaniline in Example 1 was replaced with D5 to obtain compound 6 (white powder, 29 mg) with a yield of 31%: 1 H NMR(300MHz,CD3OD)δ5.93(d,J=7.8Hz,1H),5.82(s,1H),5.78-5.67(m,3H),5.57-5.49(m,3H),5.48-5.40(m,1H).HRMS(ESI)calcd.for C 17 H 12 D5N3O[M+H] + 285.1764, found 285.1759.

[0095] Example 7

[0096] 1-(4-(ethyl-2,2-d2)phenyl)-3-(1H-indol-3-yl)urea

[0097] Synthesis of intermediate E1

[0098] D1 (977.4 mg, 2.4 mmol) and potassium carbonate (276.4 mg, 2.0 mmol) were weighed and placed in a Shrek tube. The atmosphere was replaced with argon three times, and a solution of p-nitrobenzaldehyde (300 mg, 2.0 mmol) in anhydrous tetrahydrofuran (THF) (7.5 mL) was added. The system was moved to an oil bath and reacted at 70°C for 24 hours. After completion of the reaction, the filtrate was filtered and the solvent was evaporated under reduced pressure. The filtrate was diluted with water (50 mL) and extracted with ethyl acetate (40 mL × 2). The organic phases were combined, washed with saturated brine (30 mL × 1), dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 50:1) to obtain intermediate E1 (pale yellow oily liquid, 184 mg) in a yield of 94%.

[0099] Synthesis of intermediate E2

[0100] Compound E1 (184.9 mg, 1.2 mmol) was weighed and dissolved in ethanol (2 mL). 10% palladium on carbon (40 mg) was added, and the atmosphere was replaced with hydrogen three times. The system was allowed to react at room temperature overnight. After the reaction, the mixture was filtered and the solvent was evaporated under reduced pressure to obtain a crude product containing compound E2, which was used directly in the next reaction without further purification.

[0101] Synthesis of compound 7

[0102] With reference to the synthesis method of Example 1, 4-ethylaniline in Example 1 was replaced by E2 to obtain compound 7 (white powder, 109.4 mg) with a two-step yield of 73.4%.1 H NMR(400MHz,DMSO-d6)δ10.71(s,1H),8.47(s,1H),8.39(s,1H),7.55-7.45(m,2H),7.40-7.35(m,2H),7.35-7.31(m ,1H),7.09(dd,J=8.8,7.2Hz,3H),7.04-6.94(m,1H),2.53(t,J=7.2Hz,2H),1.17-1.12(m,1H).HRMS(ESI)calcd.for C 17 H 15 D2N3O[M+H] + 282.1575found 282.1568.

[0103] Example 8

[0104] 1-(4-(ethyl-1-d)phenyl)-3-(1H-indol-3-yl)urea

[0105] Synthesis of intermediate F1

[0106] Triphenylmethylphosphonium bromide (2.2 g, 6.2 mmol) and potassium carbonate (1.4 g, 10.4 mmol) were placed in a 50 mL two-necked flask. The atmosphere was purged with argon three times, and a solution of D3 (789 mg, 2.0 mmol) in anhydrous tetrahydrofuran (THF) (15 mL) was added. The system was moved to an oil bath and reacted at 70°C for 24 hours. After completion of the reaction, the filtrate was filtered and the solvent was evaporated under reduced pressure. The filtrate was diluted with water (50 mL) and extracted with ethyl acetate (40 mL × 2). The organic phases were combined, washed with saturated brine (30 mL × 1), dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 200:1) to obtain intermediate F1 (pale yellow oily liquid, 614.1 mg) in a yield of 79%.

[0107] Synthesis of intermediate F2

[0108] Compound F1 (339.5 mg, 2.3 mmol) was weighed and dissolved in ethanol (3 mL). 10% palladium on carbon (40 mg) was added, and the atmosphere was replaced with hydrogen three times. The system was allowed to react at room temperature overnight. After the reaction, the mixture was filtered and the solvent was evaporated under reduced pressure to obtain a crude product containing compound F2, which was used directly in the next reaction without further purification.

[0109] Synthesis of compound 8

[0110] With reference to the synthesis method of Example 1, the 4-ethylaniline in Example 1 was replaced by F2 to obtain compound 8 (white powder, 219 mg) with a yield of 87%.1 H NMR(400MHz,DMSO-d6)δ10.72(s,1H),8.48(s,1H),8.40(s,1H),7.54-7.46(m,2H),7.41-7.35(m,2H),7.35-7.30 (m,1H),7.13-7.07(m,3H),7.05-6.97(m,1H),2.54(d,J=7.8Hz,1H),1.15(d,J=7.6Hz,3H).HRMS(ESI)calcd.for C 17 H 16 D4N3O[M+H] + 281.1513, found 281.1521.

[0111] Example 9

[0112] 1-(4-ethylphenyl-2-d)-3-(1H-indol-3-yl)urea

[0113] Synthesis of intermediate G1

[0114] An empty three-necked flask was protected by argon, and a solution of 1-bromo-3-ethylbenzene (926 mg, 5 mmol) in anhydrous tetrahydrofuran (THF) (10 mL) was added. n-Butyl lithium (2.5 M in THF, 3 mL) was slowly added dropwise at -78°C, and the mixture was stirred at -78°C for 1 hour. Heavy water (1 mL, D>99%) was added dropwise, and the mixture was heated to room temperature and stirred for 2 hours. Saturated sodium chloride (20 mL) was added to the reaction solution, and the mixture was extracted with dichloromethane (30 mL×3). The organic phases were combined, and the solvent was evaporated under reduced pressure in a water bath not exceeding 30°C to obtain a crude product containing compound G1, which was used directly in the next reaction without further purification.

[0115] Synthesis of compound 9

[0116] Referring to the synthesis method of Example 3, the ethylbenzene-d 10 Replacing with G1 gave compound 9 (white powder, 64 mg) with a yield of 57%: 1 H NMR(300MHz,DMSO-d6)δ10.72(s,1H),8.49(s,1H),8.41(s,1H),7.55-7.46(m,2H),7.40-7.30(m,2H) ,7.13-7.05(m,3H),7.05-6.96(m,1H),2.58-2.52(m,2H),1.16(t,J=7.6Hz,3H).HRMS(ESI)calcd.for C 17 H 16DN3O[M+H] + 281.1513, found 281.1517.

[0117] Example 10

[0118] 1-(4-(ethyl-2-d)phenyl)-3-(1H-indol-3-yl)urea

[0119] Synthesis of intermediate H1:

[0120] To a reaction flask, 4-nitrophenylacetylene (676.6 mg, 4.6 mmol) and K2CO3 (953.6 mg, 6.9 mmol) were added. Anhydrous acetonitrile (18.0 mL) was added under argon protection. The reaction was stirred at room temperature for 1.5 h, and heavy water (8.0 mL, D>99%) was added. After completion of the reaction, the mixture was diluted with water (20 mL), and the aqueous phase was extracted with ethyl acetate (30 mL x 3). The organic phases were combined, washed with saturated brine (30 mL x 1), dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether) to obtain intermediate H1 (white solid, 583.2 mg) in an 86% yield.

[0121] Synthesis of intermediate H2

[0122] Compound H1 (250 mg, 1.7 mmol) was weighed and dissolved in ethanol (5 mL). 10% palladium on carbon (80 mg) was added, and the mixture was deuterated three times. The reaction mixture was allowed to react at room temperature overnight. After the reaction, the mixture was filtered and the solvent was evaporated under reduced pressure to obtain a crude product containing compound H2, which was used directly in the next reaction without further purification.

[0123] Synthesis of compound 10

[0124] Referring to the synthesis method of Example 3, the ethylbenzene-d 10 Replaced with H2 to obtain compound 10 (white powder, 60 mg) with a yield of 34%: 1 H NMR (400MHz, DMSO-d6) δ10.74-10.70(m,1H),8.49(s,1H),8.41(s,1H),7.53-7.47(m,2H),7.40-7.35(m,2H),7. 33(d,J=8.2Hz,1H),7.09(d,3H),7.04-6.98(m,1H),2.59-2.51(m,2H),1.18-1.12(m,2H).HRMS(ESI)calcd.for C 17 H 16 DN3O[M+H] +281.1513, found 281.1517.

[0125] Example 11

[0126] 1-(4-(ethyl-2,2-d2)phenyl)-3-(1H-indol-3-yl-4,5,6-7-d4)urea

[0127] Referring to the synthesis method of Example 1, 4-ethylaniline in Example 1 was replaced with I2 to obtain compound 11 (white powder, 44 mg) with a yield of 74%: 1 H NMR (300MHz, DMSO-d6) δ10.71(s,1H),8.48(s,1H),8.41(s,1H),7.49(d,J=2.4Hz,1H),7.38(d,J= 8.5Hz,3H),7.10(d,J=8.4Hz,3H),2.58-2.52(m,2H),1.05(t,J=7.0Hz,1H).HRMS(ESI)calcd.for C 17 H 11 D6N3O[M+H] + 286.1826, found 286.1827.

[0128] Example 12

[0129] 1-(4-(ethyl-1-d)phenyl)-3-(1H-indol-3-yl-4,5,6-7-d4)urea

[0130] Referring to the synthesis method of Example 1, the 4-ethylaniline in Example 1 was replaced with F2 to obtain compound 12 (white powder, 55 mg) with a yield of 74.7%: 1 H NMR (300MHz, DMSO-d6) δ10.71(s,1H),8.48(s,1H),8.41(s,1H),7.49(d,J=2.4Hz,1H),7.41-7. 34(m,2H),7.14-7.08(m,2H),2.54(d,J=7.2Hz,1H),1.15(d,J=7.5Hz,3H).HRMS(ESI)calcd.for C 17 H 12 D5N3O[M+H] + 285.1764, found 285.1771.

[0131] Example 13

[0132] 1-(4-ethylphenyl-3-d)-3-(1H-indol-3-yl-4,5,6-7-d4)urea

[0133] Referring to the synthesis method of Example 1, 4-ethylaniline in Example 1 was replaced with M1 to obtain compound 13 (white powder, 33.9 mg) with a yield of 8%: 1 H NMR (300MHz, DMSO-d6) δ10.72(s,1H),8.49(s,1H),8.42(s,1H),7.50(d,J=2.4Hz,1H),7.42-7 .36(m,2H),7.14-7.08(m,1H),2.59-2.52(m,3H),1.16(t,J=7.6Hz,3H).HRMS(ESI)calcd.for C 17 H 12 D5N3O[M+H] + 285.11764, found 285.1760.

[0134] Example 14

[0135] 1-(4-(ethyl-1,2-d2)phenyl)-3-(1H-indol-3-yl)urea

[0136] Synthesis of intermediate J1:

[0137] 4-Nitrostyrene (273 mg, 1.83 mmol) was weighed and dissolved in ethanol (3 mL). 10% palladium on carbon (60 mg) was added, and the mixture was replaced with deuterium three times. The system was allowed to react at room temperature overnight. After the reaction was completed, the mixture was filtered, and the solvent was evaporated under reduced pressure. Water (1.5 mL) and ethanol (2 mL) were added, and the mixture was stirred at room temperature for 2 h. The mixture was diluted with water (40 mL) and extracted with ethyl acetate (30 mL × 3). The organic phases were combined, washed with saturated brine (30 mL × 1), dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to obtain a crude product containing compound J2, which was used directly in the next reaction without further purification.

[0138] Synthesis of compound 14:

[0139] Referring to the synthesis method of Example 2, B3 in Example 2 was replaced by J2 to obtain compound 14 (white powder, 69 mg) with a yield of 53.4%: 1H NMR(400MHz,CD3OD)δ7.53(d,J=7.9Hz,1H),7.41(s,1H),7.36-7.29(m,3H),7.16-7.09( m,3H),7.07-7.01(m,1H),2.58(q,J=7.0Hz,1H),1.39-0.91(m,2H).HRMS(ESI)calcd.for C 17 H 15 D2N3O[M+H] + 282.1575,found 282.1571.

[0140] Example 15

[0141] 1-(4-(ethyl-1,1,2-d3)phenyl)-3-(1H-indol-3-yl)urea

[0142] Synthesis of intermediate K1

[0143] Methyltriphenylphosphonium bromide (2.2 g, 6.2 mmol) and potassium carbonate (1.4 g, 10.4 mmol) were weighed and placed in a Shrek tube. The atmosphere was purged with argon three times. A solution of D3 (789 mg, 5.2 mmol) in anhydrous tetrahydrofuran (THF) (4 mL) was added. The system was moved to an oil bath and reacted at 70°C for 24 hours. After completion of the reaction, the filtrate was filtered and the solvent was evaporated under reduced pressure. The filtrate was diluted with water (50 mL) and extracted with ethyl acetate (40 mL × 3). The organic phases were combined, washed with saturated brine (30 mL × 1), dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 200:1) to obtain intermediate K1 (pale yellow oily liquid, 614.1 mg) in a yield of 79%.

[0144] Synthesis of intermediate K2:

[0145] Compound K1 (274.6 mg, 1.8 mmol) was weighed and dissolved in ethanol (2 mL). 10% palladium on carbon (50 mg) was added, and the mixture was replaced with deuterium three times. The reaction mixture was allowed to react at room temperature overnight. After the reaction was completed, the mixture was filtered, and the solvent was evaporated under reduced pressure. Water (1.5 mL) and ethanol (2 mL) were added, and the mixture was stirred at room temperature for 2 h. The mixture was diluted with water (40 mL) and extracted with ethyl acetate (30 mL x 3). The organic phases were combined, washed with saturated brine (15 mL), dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to obtain a crude product containing compound K2, which was used directly in the next reaction without further purification.

[0146] Synthesis of compound 15:

[0147] Referring to the synthesis method of Example 2, B3 in Example 2 was replaced by K2 to obtain compound 15 (white powder, 84.2 mg) with a yield of 85%: ( H NMR (300MHz, CDCl3) δ10.72(s,1H),8.48(s,1H),8.41(s,1H),7.53-7.46(m,2H),7.40-7.35(m,2H), 7.36-7.29(m,1H),7.13-7.06(m,3H),7.04-6.97(m,1H),1.13(d,J=5.7Hz,2H).HRMS(ESI)calcd.for C 17 H 14 D3N3O[M+H] + 283.1638, found 283.1634.

[0148] Example 16

[0149] 1-(4-(ethyl-1,2-d2)phenyl-2,3,5,6-d4)-3-(1H-indol-3-yl)urea

[0150] Synthesis of intermediate L2:

[0151] Compound L1 (450.2 mg, 2.94 mmol) was weighed and dissolved in deuterated methanol (2 mL). 10% palladium on carbon (80 mg) was added and the deuterium gas was replaced three times. The system was brought to room temperature and reacted overnight. After the reaction was completed, the mixture was filtered and the solvent was evaporated under reduced pressure. Water (1.5 mL) and anhydrous ethanol (2 mL) were added and stirred at room temperature for 2 h. The mixture was diluted with water (40 mL) and extracted with ethyl acetate (30 mL × 3). The organic phases were combined and washed with saturated brine (30 mL × 1), dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to obtain a crude product containing compound L1, which was used directly in the next reaction without further purification.

[0152] Synthesis of compound 16

[0153] Referring to the synthesis method of Example 2, B3 in Example 2 was replaced by L1 to obtain compound 16 (white powder, 129.4 mg) with a yield of 78%: 1 H NMR(300MHz,CD3OD)δ7.55-7.50(m,1H),7.41(s,1H),7.37-7.32(m,1H),7.17-7.1 0(m,1H),7.07-7.00(m,1H),2.57(s,1H),1.22-1.12(m,2H).HRMS(ESI)calcd.for C 17 H 11D6N3O[M+H] + 286.1826, found 286.1819.

[0154] Example 17

[0155] 1-(4-(ethyl-1,2-d2)phenyl-2,3,5,6-d4)-3-(1H-indol-3-yl-4,5,6-7-d4)urea

[0156] Referring to the synthesis method of Example 1, the 4-ethylaniline in Example 1 was replaced with L2 to obtain compound 17 (white powder, 61 mg) with a yield of 70%: 1 H NMR(400MHz,CD3OD)δ7.41(s,1H),2.62-2.51(m,1H),1.19(t,J=7.3Hz,2H).HRMS(ESI)calcd.for C 17 H7D 10 N3O[M+H] + 290.2078,found 290.2072.

[0157] Example 18

[0158] 1-(4-(ethyl-1,1,2-d3)phenyl)-3-(1H-indol-3-yl-4,5,6-7-d4)urea

[0159] Referring to the synthesis method of Example 1, the 4-ethylaniline in Example 1 was replaced with K2 to obtain compound 18 (white powder, 60 mg) with a yield of 70%: 1 H NMR(300MHz,CD3OD)δ7.41(s,1H),7.32(d,J=8.5Hz,2H),7.12(d,J=8.5Hz,2H),1.18(d,J=5.6Hz,2H).HRMS(ESI)calcd.for C 17 H 10 D7N3O[M+H] + 287.1889, found 287.1881.

[0160] Example 19

[0161] 1-(4-(ethyl-1,2-d2)phenyl)-3-(1H-indol-3-yl-4,5,6-7-d4)urea

[0162] Referring to the synthesis method of Example 1, 4-ethylaniline in Example 1 was replaced with J1 to obtain compound 19 (white powder, 58 mg) with a yield of 68%: 1 H NMR (400MHz, CD3OD) δ7.41(s,1H),7.32(d,J=8.4Hz,2H),7.12(d,J=8.5Hz,2H),2.58(q,J=7.2Hz,1H),1.22-1.14(m,2H).HRMS(ESI)calcd.for C 17 H 11 D6N3O[M+H] + 286.1826, found 286.1816.

[0163] Example 20

[0164] 1-(4-(ethyl-2-d)phenyl)-3-(1H-indol-3-yl-4,5,6-7-d4)urea

[0165] Referring to the synthesis method of Example 1, the 4-ethylaniline in Example 1 was replaced by H2 to obtain compound 20 (white powder, 50 mg) with a yield of 59%: 1 H NMR(400MHz,CD3OD)δ7.41(s,1H),7.32(d,J=8.4Hz,2H),7.12(d,J=8.4Hz,2H),2.59(q,J=7.3Hz,2H),1.24-1.16(m,2H).HRMS(ESI)calcd.for C 17 H 12 D5N3O[M+H] + 285.1764, found 285.1767.

[0166] Example 21

[0167] 1-(4-ethylphenyl-3-d)-3-(1H-indol-3-yl)urea

[0168] Synthesis of intermediate M1:

[0169] An empty three-necked flask was protected by argon, and a solution of 1-bromo-2-ethylbenzene (1.4 g, 7.5 mmol) in anhydrous tetrahydrofuran (THF) (15 mL) was added. n-Butyl lithium (2.5 M in THF, 4.5 mL) was slowly added dropwise at -78 ° C. and stirred at -78 ° C for 1 hour. Heavy water (3 mL, D>99%) was added dropwise. After warming to room temperature and stirring for 2 hours, saturated sodium chloride (15 mL) was added to the reaction solution, and extracted with dichloromethane (30 mL×3). The organic phases were combined and the solvent was evaporated under reduced pressure in a water bath not higher than 30 ° C to obtain a crude product containing compound M1, which was used directly in the next reaction without further purification.

[0170] Synthesis of compound 21:

[0171] Referring to the synthesis method of Example 3, the ethylbenzene-d 10 Replaced with M1 to obtain compound 21 (white powder, 50 mg) with a yield of 46%: 1 H NMR(400MHz,CD3OD)δ7.53(d,J=7.9Hz,1H),7.41(s,1H),7.36-7.29(m,3H),7.15-7.09(m, 2H),7.06-7.01(m,1H),2.59(q,J=7.6Hz,2H),1.21(t,J=7.6Hz,3H).HRMS(ESI)calcd.for C 17 H 16 DN3O[M+H] + 281.1513, found 281.1519.

[0172] Example 22

[0173] 1-(4-ethylphenyl-2,6-d2)-3-(1H-indol-3-yl)urea

[0174] Synthesis of intermediate N1:

[0175] An empty three-necked flask was protected by argon, and a solution of 3,5-dibromoethylbenzene (2 g, 7.6 mmol) in anhydrous tetrahydrofuran (THF) (15 mL) was added. n-Butyl lithium (2.5 M in THF, 9.2 mL) was slowly added dropwise at -78°C, and the mixture was stirred at -78°C for 1 hour. Heavy water (8 mL, D>99%) was added dropwise, and the mixture was heated to room temperature and stirred for 2 hours. Saturated sodium chloride (15 mL) was added to the reaction solution, and the mixture was extracted with dichloromethane (40 mL×3). The organic phases were combined, and the solvent was evaporated under reduced pressure in a water bath not exceeding 30°C to obtain a crude product containing compound N1, which was used directly in the next reaction without further purification.

[0176] Synthesis of compound 22:

[0177] Referring to the synthesis method of Example 3, the ethylbenzene-d 10 Replaced with N1 to obtain compound 22 (white powder, 51 mg) with a yield of 67%: 1 H NMR(400MHz,CD3OD)δ7.53(d,J=8.0Hz,1H),7.41(s,1H),7.36-7.33(m,1H),7.15-7.09(m, 3H),7.06-7.01(m,1H),2.59(q,J=7.6Hz,2H),1.21(t,J=7.6Hz,3H).HRMS(ESI)calcd.for C 17 H 15 D2N3O[M+H] + 282.1575, found 282.1581.

[0178] Example 23

[0179] 1-(4-ethylphenyl-2,6-d2)-3-(1H-indol-3-yl-4,5,6-7-d4)urea

[0180] Referring to the synthesis method of Example 1, 4-ethylaniline in Example 1 was replaced by N1 to obtain compound 23 (white powder, 33 mg) with a yield of 31%: 1 H NMR(300MHz,CD3OD)δ7.41(s,1H),7.12(s,2H),2.59(q,J=7.6Hz,2H),1.21(t,J=7.6Hz,3H).HRMS(ESI)calcd.for C 17 H 11 D6N3O[M+H] + 286.1862, found 286.1832.

[0181] Example 24

[0182] 1-(4-ethylphenyl-3-d)-3-(1H-indol-3-yl-4,5,6-7-d4)urea

[0183] Referring to the synthesis method of Example 1, 4-ethylaniline in Example 1 was replaced with M1 to obtain compound 24 (white powder, 47 mg) with a yield of 34%: 1H NMR(400MHz,CD3OD)δ7.41(s,1H),7.33-7.28(m,1H),7.16-7.06(m,2H),2.59(q,J=7.6Hz,2H),1.21(t,J=7.6Hz,3H).HRMS(ESI)calcd.for C 17 H 12 D5N3O[M+H] + 285.1764, found 285.1772.

[0184] Example 25

[0185] 1-(4-Ethylphenyl)-3-(1H-indol-3-yl-2d)urea

[0186] Synthesis of intermediate O1:

[0187] An empty three-necked flask was protected by argon, and a solution of N-benzenesulfonic acid indole (1 g, 3.9 mmol) in anhydrous tetrahydrofuran (THF) (20 mL) was added. n-Butyl lithium (2.5 M in THF, 3.2 mL) was slowly added dropwise at -78°C, and the mixture was stirred at -78°C for 1 hour. Heavy water (1 mL, D>99%) was added dropwise. After warming to room temperature and stirring for 2 hours, saturated sodium chloride (15 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (40 mL×3). The organic phases were combined, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 50 / 1) to obtain O1 (white solid, 900 mg) in a yield of 89%.

[0188] Synthesis of intermediate O2:

[0189] To O1 (686.2 mg, 2.66 mmol) was added ethanol (20 mL), and 2 M NaOH solution (8.7 mL) was slowly added dropwise. The mixture was heated under reflux at 65°C. After the reaction was complete, the mixture was diluted with water (20 mL) and extracted with ethyl acetate (30 mL × 3). The organic phases were combined, washed with saturated brine (30 mL × 1), dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 50 / 1) to obtain O2 (white solid, 312 mg) in a yield of 99%.

[0190] Synthesis of compound 25:

[0191] Referring to the synthesis method of Example 1, A4 in Example 1 was replaced by O3 to obtain compound 25 (white powder, 73 mg) with a yield of 73%: 1H NMR(400MHz,CD3OD)δ7.54-7.51(m,1H),7.36-7.29(m,3H),7.15-7.10(m,3H),7.0 6-7.01(m,1H),2.59(q,J=7.6Hz,2H),1.21(t,J=7.6Hz,3H).HRMS(ESI)calcd.for C 17 H 16 DN3O[M+H] + 281.1513, found 281.1519.

[0192] Example 26

[0193] 1-(4-Ethylphenyl)-3-(1H-indol-3-yl-4-d)urea

[0194] Synthesis of intermediate P1:

[0195] An empty three-necked flask was protected by argon, and a solution of 4-bromo-1-(tert-butyldimethylsilyl)indole (369 mg, 1.2 mmol) in anhydrous tetrahydrofuran (THF) (5.5 mL) was added. n-Butyl lithium (2.5 M in THF, 0.92 mL) was slowly added dropwise at -78°C. The mixture was stirred at -78°C for 1 hour, and heavy water (0.4 mL, D>99%) was added dropwise. After warming to room temperature and stirring for 2 hours, saturated sodium chloride (15 mL) was added to the reaction solution. The mixture was extracted with ethyl acetate (40 mL×3). The organic phases were combined, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether) to obtain P1 (colorless oily liquid, 120 mg) with a yield of 80%.

[0196] Synthesis of intermediate P2:

[0197] To a solution of P1 (360 mg, 1.6 mmol) in anhydrous tetrahydrofuran (THF) (18 mL) was added tetrabutylammonium fluoride (TBAF) (1 M in THF, 5 mL) (376.5 mg, 1.4 mmol), and the mixture was stirred at room temperature. After the reaction was complete, the reaction solution was diluted with water (20 mL) and extracted with ethyl acetate (30 mL × 3). The organic phases were combined, washed with saturated brine (30 mL × 1), dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 40 / 1) to obtain P2 (a colorless oily liquid, 48 mg) in a yield of 78%.

[0198] Referring to the synthesis method of Example 1, A4 in Example 1 was replaced with P2 to obtain compound 26 (white powder, 56 mg) with a yield of 56%: 1H NMR(400MHz,CD3OD)δ7.41(s,1H),7.37-7.27(m,3H),7.15-7.08(m,3H),7.07-7.00(m,1H),2.59(q,J=7.6Hz,2H),1.21(t,J=7.6Hz,3H).HRMS(ESI)calcd.for C 17 H 16 DN3O[M+H] + 281.1513,found 281.1513.

[0199] Example 27

[0200] Evaluation of the inhibitory activity of compounds on the STING signaling pathway in THP1-Dual cells

[0201] Experimental Principle: THP1-Dual is a commercially available dual-reporter cell line for NF-κB and IRF signaling pathways. The transcriptional activity of the IRF signaling pathway can be assessed by detecting secreted luciferase. This cell line also expresses cGAS and STING proteins. Introduction of exogenous double-stranded DNA can activate the cGAS-STING signaling pathway, thereby enhancing IRF transcriptional activity. Therefore, exogenous HT-DNA is transfected to activate the cGAS-STING-IRF signaling pathway within the cells. After adding compounds to interfere with the STING signaling pathway, the inhibitory activity of the compounds on STING is assessed by detecting secreted luciferase.

[0202] Experimental reagents and materials: heat-inactivated serum (Biological Industries), 1640 culture medium (Biological Industries), penicillin-streptomycin dual antibody (Biological Industries), THP1-Dual TM Cells (InvivoGen), HT-DNA (Sigma Aldrich, prepared as a 2.5 mg / mL stock solution), Opti-MEM (Gibco), Lipo6000 (Biyuntian), QUANTI-Luc TM (InvivoGen).

[0203] Experimental method: (1) Cell seeding: Centrifuge, resuspend and count THP1-Dual cells in good growth condition. Take some cells and mix them with pre-prepared HT-DNA working solution (for example, mix 1μg of HT-DNA storage solution with 2μL of Lipo6000 in 0.5mL of Opti-MEM and let it stand at room temperature for 10 minutes before use). Prepare a culture medium solution with an HT-DNA concentration of 0.5μg / mL and a cell concentration of 800,000 / mL. Take 100μL and add it to the 96-well plate as the experimental well; and add the corresponding cell solution without HT-DNA to the control well. (2) Cell administration: Prepare the test compound into a 10mM storage solution and dilute it with culture medium to 20, 4, 0.8, 0.16, 0.008, 0.0064, and 0.00128μM solutions. Add 100μL of the solution to the experimental wells in sequence, and add 100μL of culture medium to the control wells and model wells. Place in an incubator and incubate for 16 to 18 hours before testing. (3) Testing: Place QUANTI-Luc TM The powder was prepared into a detection solution with purified water according to the instructions, and stored at 4°C after packaging. Add 10 μL of detection solution to a white opaque 384-well plate. Take out the 96-well plate and centrifuge it at 1000 rpm for 1 minute using a 96-well plate centrifuge. Then, take 4 μL of the supernatant in turn and add it to the 96-well plate with the detection solution. The chemiluminescence was detected using an enzyme-labeled instrument. (4) Data processing: The inhibition rate of the compound at a fixed concentration was calculated according to the following formula: The inhibition rate of the compound at a certain concentration = 1-(chemiluminescence value of the compound well at a certain concentration - chemiluminescence value of the control well) / (chemiluminescence value of the model well - chemiluminescence value of the control well) × 100%; then, a curve was fitted based on the inhibition rate of the compound at each concentration to calculate the half inhibition rate (IC) of the compound. 50 value).

[0204] Experimental results: The inhibitory activity of the compounds on the STING signaling pathway in THP-1 Dual cells is shown in Table 2.

[0205] Table 2. Inhibitory activity of compounds on the STING signaling pathway in THP-1 Dual cells

[0206] The experimental results (Table 2) show that the compounds represented by formula I of the present invention can significantly inhibit the activation of the STING signaling pathway in THP1-Dual cells, and the IC values ​​of most compounds for STING inhibition are 50 The values ​​were in the nanomolar range, and most compounds were more active than the positive control compound H-151, with compounds 6 and 21 being significantly more active than H-151. This suggests that the compounds of Formula I of the present invention can effectively inhibit the activation of the STING signaling pathway.

[0207] Example 28

[0208] Study on the metabolic stability of compounds in human liver microsomes:

[0209] The stability evaluation of human liver microsomes is an important means of preclinical evaluation of the pharmacokinetic properties of candidate compounds in drug development. The experimental incubation system (volume 250 μL, n = 3) consists of liver microsomes, test substance working solution and phosphate buffer. The incubation system is incubated at 37 ° C for one hour. The timing starts after the addition of NADPH solution. The reaction is terminated by adding the stop solution at each time point. The sampling intervals are 0, 5, 15, 30, and 60 minutes, for a total of 5 points. The negative control does not add NADPH, and the sampling time points are 0 and 60 minutes. LC-MS / MS is used for analysis, and the absolute value of the slope k is measured by plotting the natural logarithm of the percentage of the remaining amount of the test substance against time, and calculated according to the following formula: T 1 / 2 (Half-life) = ln2 / k = 0.693 / k; Clint (μL / min / mg protein) (Clearance) = Ln(2)*1000 / T 1 / 2 / Protein Conc. The experimental results are shown in Table 3.

[0210] Table 3. Metabolic half-life of compounds in human liver microsomes (T 1 / 2 ) and human liver microsomal clearance

[0211] The experimental results showed that compared with the positive control compound H-151, most of the compounds T 1 / 2 The metabolic stability of compounds 2, 4, 8 and 15 was significantly stronger than that of H-151, which showed that the deuteration strategy adopted in the present invention did improve the metabolic stability of the compounds, suggesting that the compounds of the present invention may have better pharmacokinetic properties.

[0212] Example 29

[0213] Effect of compound 15 on the mouse psoriasis model induced by imiquimod cream

[0214] In order to verify the effect of the compound of the present invention on autoimmune diseases, the psoriasis model of mice induced by imiquimod cream was used to verify the efficacy of the compound.

[0215] Experimental animals: 8-week-old female Balb / c mice were purchased from Beijing Weitonglihua Laboratory Animal Co., Ltd.

[0216] Test drug preparation: Heat 7.2g of PEG400 to 55-60°C, add 200mg of the test drug, stir, and dissolve using an ultrasonic cleaner. Invert and mix several times. Once dissolved, heat 2.8g of PEG3350 to 55-60°C to dissolve, then mix thoroughly with the PEG400-test drug mixture. Dispense into 1g tubes to prepare a 2% test drug ointment. Apply 0.2g / mouse to the back of each mouse. Simultaneously prepare a control ointment containing no drug.

[0217] Modeling and drug administration: Mice were randomly divided into a blank group, a model group, and a compound 15 group (applied with 2% 15 ointment), with 5 mice in each group. The mice were depilated on their backs to expose a 2cm×3cm skin area. After depilation, they were allowed to adapt for 2 days. 5% imiquimod (IMQ) cream 50 mg / mouse was applied to the back once a day for 7 days of modeling. At the same time, the compound 15 group was applied with 2% compound 15 ointment, and the control and model group animals were applied with a drug-free control ointment, 0.2 g / mouse, once a day, for a total of 7 days. The mice were weighed every day, photos of the back were taken, and PASI (psoriasis area and severity index) scores were performed.

[0218] The experimental results (Figure 1) show that after applying imiquimod cream, psoriasis-like pathological phenotypes such as erythema, thickening and squama are significantly seen on the back of mice, while after applying compound 15 ointment, this pathological feature can be significantly improved. At the same time, according to the PASI score (Figure 2), it can also be seen that compound 15 can significantly reduce the psoriasis-like inflammatory damage, skin thickening and squama on the back of model mice. This shows that compound 15 has significant immunomodulatory efficacy and can be used for the treatment of psoriasis. It further suggests that it can be used to prevent and treat infectious diseases, inflammatory diseases, autoimmune diseases, metabolic diseases, organ fibrosis diseases, cardiovascular and cerebrovascular diseases, respiratory diseases, central nervous system diseases, cancer or precancerous syndromes. Other compounds of the present invention also have similar therapeutic effects.

[0219] Example 30

[0220] Therapeutic effects of compounds 6, 15, 21 and benvimod on the mouse psoriasis model induced by imiquimod cream

[0221] Experimental animals: 8-week-old female Balb / c mice were purchased from Beijing Weitonglihua Laboratory Animal Co., Ltd.

[0222] Test drug preparation: Heat 7.2g of PEG400 to 55-60°C, add 200mg of the test drug, stir, and dissolve using an ultrasonic cleaner, repeatedly inverting and mixing. Once dissolved, heat 2.8g of PEG3350 to 55-60°C to dissolve, then mix thoroughly with the PEG400-mixed test drug. Dispense into 1g tubes to prepare a 2% test drug ointment. Apply 0.2g / mouse to the back of the mouse. A control ointment containing no drug is also prepared. Modeling and Dosing: Mice were randomly divided into a blank group, a model group, a compound 6 group (applied with 2% ointment of compound 6), a compound 15 group (applied with 2% ointment of compound 15), a compound 21 group (applied with 2% ointment of compound 21), and a belimud group (applied with 2% ointment of belimud), with 5 mice per group. The backs of the mice were depilated, exposing a 2cm x 3cm skin area. The mice were allowed to acclimate for 2 days after depilation. 5% Imiquimod (IMQ) cream 50 mg / animal was applied to the back once daily for 7 days. At the same time, the drug-treated group was treated with an ointment containing the corresponding drug, while the control and model groups were treated with a control ointment containing no drug, 0.2 g / animal, once daily for 7 days. The animals were weighed daily, their backs photographed, and PASI (Psoriasis Area and Severity Index) scores were performed. Benvimod is a psoriasis treatment drug, and the API was purchased from Jiangxi Ruiweier Biotechnology Co., Ltd.

[0223] The experimental results (Figure 3) show that after applying imiquimod cream, psoriasis-like pathological phenotypes such as erythema, thickening and scaling are significantly seen on the back of mice, while after applying compound 6, 15 and 21 ointment, this pathological feature can be significantly improved. At the same time, according to the PASI score (Figure 4), it can be seen that compounds 6, 15 and 21 can significantly reduce psoriasis-like inflammatory damage, skin thickening and scaling on the back of model mice; the therapeutic effects of these three compounds are comparable to benvimod, and the effect of compound 21 is slightly better than that of benvimod. This shows that compounds 6, 15 and 21 have significant immunomodulatory effects and can be used for the treatment of psoriasis, and further suggests that they can be used to prevent and treat infectious diseases, inflammatory diseases, autoimmune diseases, metabolic diseases, organ fibrosis diseases, cardiovascular and cerebrovascular diseases, respiratory diseases, central nervous system diseases, cancer or precancerous syndromes and other diseases. Other compounds of the present invention also have similar therapeutic effects.

[0224] Example 31

[0225] tablet

[0226] The compound obtained in any one of Examples 1-26 (50 g), hydroxypropyl methylcellulose E (150 g), starch (200 g), appropriate amount of povidone K30 and magnesium stearate (1 g) were mixed, granulated and tableted.

[0227] ointment

[0228] Heat 7.2 g of PEG400 to 55-60° C., add 200 mg of any of the compounds prepared in Examples 1-26, stir, and dissolve using an ultrasonic cleaner. Invert and mix repeatedly several times. After dissolution, heat 2.8 g of PEG3350 to 55-60° C., dissolve, and mix with the PEG400 mixed with the test drug. Dispense into 1 g / tube to prepare a 2% test drug ointment.

Claims

1. A deuterated indole urea compound as shown in the following formula I or a pharmaceutically acceptable salt, solvate or prodrug thereof: in, Y 1 ,Y 2 ,Y 3 ,Y 4 ,Y 5 ,Y 6 ,Y 7 ,Y 8 ,Y 9 ,Y 10 ,Y 11 ,Y 12 ,Y 13 or Y 14 are each independently selected from hydrogen or deuterium; and Y 1 ,Y 2 ,Y 3 ,Y 4 ,Y 5 ,Y 6 ,Y 7 ,Y 8 ,Y 9 ,Y 10 ,Y 11 ,Y 12 ,Y 13 or Y 14 At least one of them is deuterium.

2. The deuterated indole urea compound or a pharmaceutically acceptable salt, solvate or prodrug thereof according to claim 1, characterized in that: The Y 1 , Y 2 , Y 3 , Y 4 , Y 5 , Y 6 , Y 7 , Y 8 , Y 9 , Y 10 , Y 11 , Y 12 , Y 13 or Y 14 are each independently selected from hydrogen or deuterium; and Y 6 , Y 7 , Y 8 , Y 9 , Y 10 , Y 11 , Y 12 , Y 13 or Y 14 At least one of them is deuterium.

3. The deuterated indole urea compound or a pharmaceutically acceptable salt, solvate or prodrug thereof according to claim 2, characterized in that: The Y 1 , Y 2 , Y 3 , Y 4 , Y 5 , Y 6 , Y 7 , Y 8 , Y 9 , Y 10 , Y 11 , Y 12 , Y 13 or Y 14 are each independently selected from hydrogen or deuterium; and Y 10 , Y 11 , Y 12 , Y 13 or Y 14 At least one of them is deuterium.

4. The deuterated indole urea compound or a pharmaceutically acceptable salt, solvate or prodrug thereof according to claim 1, characterized in that: The compound or its pharmaceutically acceptable salt is selected from any one of the following:

5. Use of a deuterated indole urea compound or a pharmaceutically acceptable salt, solvate or prodrug thereof according to any one of claims 1 to 4 in the preparation of a STING inhibitor.

6. Use of a deuterated indole urea compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt, solvate or prodrug thereof in the preparation of a medicament for preventing or treating a STING-mediated disease.

7. The use according to claim 6, characterized in that The STING-mediated diseases include infectious diseases, inflammatory diseases, autoimmune diseases, metabolic diseases, organ fibrosis diseases, cardiovascular and cerebrovascular diseases, respiratory diseases, central nervous system diseases, cancer or precancerous syndromes.

8. Use of the deuterated indole urea compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt, solvate or prodrug thereof in the preparation of an immune adjuvant drug.

9. A pharmaceutical composition for preventing or treating a STING-mediated disease, comprising a compound according to any one of claims 1 to 4 or a pharmaceutically acceptable salt, solvate or prodrug thereof as an active ingredient and a pharmaceutically acceptable carrier.

10. The pharmaceutical composition according to claim 9, characterized in that The pharmaceutical composition is preferably a capsule, powder, tablet, granule, pill, injection, syrup, oral solution, inhalant, ointment, solution, cream, gel, powder, lotion, tincture, suppository or patch.

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