Selective TYK2 inhibitor compound as well as preparation method and application thereof

Abstract

The invention provides a selective TYK2 inhibitor compound as well as a preparation method and application thereof, and belongs to the technical field of medicinal chemistry and pharmacotherapeutics. The invention provides the aryl pyrimidinamine compound with a brand new structure, and the aryl pyrimidinamine compound has good safety and good TYK2 and JAK inhibitory activity, and provides a new choice for preparing medicines for preventing or treating autoimmune diseases.

Description

Technical Field

[0001] The present invention relates to the technical fields of medicinal chemistry and pharmacotherapeutics, and particularly relates to a selective TYK2 inhibitor compound, a preparation method thereof, and an application thereof. Background Art

[0002] Ulcerative Colitis (UC) is a chronic non-specific intestinal inflammatory disease that mainly affects the mucosal layer of the colon and rectum, and its main feature is continuous and diffuse inflammation of the colorectal mucosa. In recent years, the incidence of ulcerative colitis has shown an increasing trend globally. According to epidemiological data statistics, the incidence of ulcerative colitis in developed countries has reached as high as 0.1%, and in developing countries, it also shows an obvious growth trend. The pathogenesis of ulcerative colitis is complex and involves multiple factors such as genetics, immunity, and environment. Patients usually present with symptoms such as recurrent diarrhea, abdominal pain, and mucopurulent bloody stools, which seriously affect the quality of life. At the same time, ulcerative colitis may also increase the risk of colon cancer, posing long-term health hazards to patients. In terms of treatment, there is no radical cure for ulcerative colitis, and the condition is mainly controlled by drugs, symptoms are relieved, complications are prevented and treated, and canceration is prevented. Currently, the therapeutic drugs for ulcerative colitis in clinical practice have the risks of easy recurrence, drug dependence, drug resistance, and accumulation of adverse events. Due to the chronic and recurrent characteristics of ulcerative colitis, patients need to take drugs for a long time or even for life, bringing a heavy economic burden to society and families. Therefore, finding safe and effective drugs for treating ulcerative colitis with novel mechanisms of action has become an urgent problem to be solved in the current medical field.

[0003] Studies have found that, in normal and pathological states, the JAK-STAT signaling pathway plays an important role in intracellular signal transduction in various cell processes. TYK2 affects the phosphorylation levels of STAT-1 and STAT-2 by regulating the signaling pathways downstream of the IL-12, IL-23, and type I IFNs receptors. IL-23 and IL-12 are considered to be key cytokines for initiating and maintaining chronic inflammation, and tyrosine kinase 2 (TYK2) is the key intracellular signal transduction link between them. Inhibiting the activity of TYK2 can cut off the connection between IL23 / IL-12. As an intracellular signal kinase, TYK2 plays a key role in mediating the signal transduction of IL-23, IL-12, and type I interferon (IFN). In recent years, the potential of TYK2 inhibitors in treating ulcerative colitis (UC) has received extensive attention. TYK2 inhibitors specifically inhibit the activity of TYK2, thereby blocking the conduction of downstream inflammatory signals, controlling the development of inflammation, and thus playing a role in treating ulcerative colitis.

[0004] This mechanism of action is more precise and efficient compared to traditional anti-inflammatory drugs. The mechanism of TYK2 inhibitors in the treatment of ulcerative colitis is significantly innovative, bringing new hope and breakthroughs to the treatment of ulcerative colitis. Therefore, designing highly selective inhibitors targeting TYK2 is of great significance for the treatment and mechanism research of ulcerative colitis. Summary of the Invention

[0005] In view of the above deficiencies in the prior art, the present invention provides a compound with a novel structure that selectively inhibits TYK2, as well as its preparation method and applications.

[0006] To achieve the above object, the specific technical solutions of the present invention are as follows:

[0007] In the first aspect, the present invention provides a selective TYK2 inhibitor compound having the structure shown in Formula I:

[0008]

[0009] In Formula I, X is -NH-(C=O)- or -NH-(CH2)-;

[0010] R1 is a heteroaryl or a substituted aryl, and the substituents of the aryl include, but are not limited to, halogen (including F, Cl, Br, or I), trifluoromethyl, methoxy, hydroxy, or methyl;

[0011] R2 is an alkane (including cycloalkane and straight-chain alkane), aryl, or a substituted aryl, and the substituents of the aryl include, but are not limited to, halogen (including F, Cl, Br, or I), trifluoromethyl, methoxy, hydroxy, or methyl.

[0012] For each of the above variables, any combination and collocation other than the above groups are also contemplated herein. It is understood that the substituents and substitution patterns on the compounds provided herein can be selected by those skilled in the art in order to provide compounds that are chemically stable and can be synthesized using techniques known in the art and the techniques described herein.

[0013] Preferably, the compound has the structure shown in Formula I-a:

[0014]

[0015] In Formula I-a, R1 is pyridine or an aryl substituted at least at one of the ortho, meta, or para positions, and the substituents of the aryl are halogen (including F, Cl, Br, or I), trifluoromethyl, methoxy, hydroxy, or methyl; R2 is an alkane (including cycloalkane and straight-chain alkane), aryl, or a substituted aryl, and the substituents of the aryl are halogen (including F, Cl, Br, or I), trifluoromethyl, methoxy, hydroxy, or methyl.

[0016] Preferably, the compound has the structure shown in Formula I-b:

[0017]

[0018] In Formula I-b, R1 is 2-fluorophenyl, 2-chlorophenyl, 2-fluoro-6-chlorophenyl or 2,6-dimethylphenyl; R2 is an alkane (including cycloalkane and straight-chain alkane), aryl or substituted aryl, and the substituents of the aryl are halogen (including F, Cl, Br or I), trifluoromethyl, methoxy, hydroxy or methyl.

[0019] Preferably, R2 has one of the following structures:

[0020]

[0021] Preferably, the compound has one of the following structures:

[0022]

[0023]

[0024] In a second aspect, the present invention provides a method for preparing the compound. When X is -NH-(CH2)-, the compound is prepared through the following reaction route:

[0025]

[0026] The reaction process is as follows: The starting material (SM) reacts with R1-CH2-NH2 through a substitution reaction to obtain Intermediate 1; Intermediate 1 reacts with tert-butyl (4-aminophenyl)carbamate through a substitution reaction to obtain Intermediate 2; Intermediate 2 is deprotected by removing the BOC group to obtain Intermediate 3; Intermediate 3 is acylated to obtain the target compound I-a;

[0027] When X is -NH-(C=O)-, the compound is prepared through the following reaction route:

[0028]

[0029] The reaction process is as follows: The starting material (SM) is acylated to obtain Intermediate 4; Intermediate 4 reacts with tert-butyl (4-aminophenyl)carbamate through a substitution reaction to obtain Intermediate 5; Intermediate 5 is deprotected by removing the BOC group to obtain Intermediate 6; Intermediate 6 is acylated to obtain the target compound I-b.

[0030] In a third aspect, the present invention provides the use of the compound or its pharmaceutically acceptable salt, solvate, ester, acid, metabolite or prodrug in the preparation of a drug for a TYK2 inhibitor or a JAK inhibitor.

[0031] Preferably, the JAK includes JAK1, JAK2, and JAK3.

[0032] In a fourth aspect, the present invention provides the use of the compound or its pharmaceutically acceptable salt, solvate, ester, acid, metabolite, or prodrug in the preparation of a drug for preventing or treating autoimmune diseases, including but not limited to rheumatoid arthritis, psoriasis, Crohn's disease, inflammatory bowel disease, ulcerative colitis, or autoimmune-related skin diseases.

[0033] Preferably, the autoimmune-related skin diseases include alopecia areata, vitiligo, lupus erythematosus, lichen planus, lichen nitidus, lichen sclerosus et atrophicus, panniculitis, atopic dermatitis, eczema, or neurodermatitis.

[0034] In a fifth aspect, the present invention provides a pharmaceutical composition comprising the compound or its pharmaceutically acceptable salt, solvate, ester, acid, metabolite, or prodrug, and one or more pharmaceutically acceptable carriers, diluents, or excipients.

[0035] Compared with the prior art, the advantages of the present invention are as follows:

[0036] A novel structure of a JAK inhibitor, its preparation method, pharmaceutical composition, and application

[0037] The present invention provides an arylpyrimidineamine compound with a novel structure. In in vitro cell experiments, the arylpyrimidineamine compound has a good inhibitory effect on TYK2 in the JAK-STAT signaling pathway. The arylpyrimidineamine compound or its pharmaceutically acceptable salt can be used to prepare a drug for preventing or treating ulcerative colitis, has a good preventive or therapeutic effect, and has certain drug-forming properties. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 It is the experimental result of the effect of compound I-13 on the JAK-STAT signaling pathway. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0039] The technical solutions of the present invention will be described clearly and completely below. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art without creative efforts based on the embodiments of the present invention belong to the scope of protection of the present invention.

[0040] The present invention provides a selective TYK2 inhibitor compound having the structure shown in Formula I:

[0041]

[0042] In Formula I, X is -NH-(C=O)- or -NH-(CH2)-;

[0043] R1 is a heteroaryl or a substituted aryl, and the substituents of the aryl include but are not limited to halogens (including F, Cl, Br or I), trifluoromethyl, methoxy, hydroxy or methyl;

[0044] R2 is an alkane (including cycloalkane and straight-chain alkane), an aryl or a substituted aryl, and the substituents of the aryl include but are not limited to halogens (including F, Cl, Br or I), trifluoromethyl, methoxy, hydroxy or methyl.

[0045] For each of the above variables, any combination and arrangement other than the above groups are also contemplated herein. It is understood that the substituents and substitution patterns on the compounds provided herein can be selected by those skilled in the art so as to provide compounds that are chemically stable and can be synthesized using techniques known in the art and the techniques described herein.

[0046] In some examples, when X is -NH-(CH2)-, R1 is pyridine or an aryl substituted at least at one of the ortho, meta or para positions, and the substituents of the aryl are halogens (including F, Cl, Br or I), trifluoromethyl, methoxy, hydroxy or methyl; R2 is an alkane (including cycloalkane and straight-chain alkane), an aryl or a substituted aryl, and the substituents of the aryl are halogens (including F, Cl, Br or I), trifluoromethyl, methoxy, hydroxy or methyl. When X is -NH-(C=O)-, R1 is 2-fluorophenyl, 2-chlorophenyl, 2-fluoro-6-chlorophenyl or 2,6-dimethylphenyl; R2 is an alkane (including cycloalkane and straight-chain alkane), an aryl or a substituted aryl, and the substituents of the aryl are halogens (including F, Cl, Br or I), trifluoromethyl, methoxy, hydroxy or methyl.

[0047] In some examples, R2 has one of the following structures:

[0048]

[0049] In some examples, the arylpyrimidine compound has one of the following structures:

[0050]

[0051]

[0052] In the following specific examples, when X is -NH-(CH2)-, the compound is prepared by the following reaction route:

[0053]

[0054] The reaction conditions are as follows: (a) ethanol, 0 - 25 °C; (b) HCl, 1-butanol, 80 °C; (c) CF3COOH, DCM, RT; (d) Et3N, DCM, 0 °C - RT;

[0055] The reaction process is as follows: The starting material (SM) reacts with R1-CH2-NH2 through a substitution reaction to obtain intermediate 1; Intermediate 1 reacts with tert-butyl (4-aminophenyl)carbamate through a substitution reaction to obtain intermediate 2; Intermediate 2 removes the BOC protecting group to obtain intermediate 3; Intermediate 3 is acylated to obtain the target compound I-a.

[0056] When X is -NH-(C=O)-, the compound is prepared through the following reaction route:

[0057]

[0058] The reaction conditions are as follows: (a) NaH, DMF, 0 °C - RT; (b) HCl, 1-butanol, 80 °C; (c) CF3COOH, DCM, RT; (d) Et3N, DCM, 0 °C - RT;

[0059] The reaction process is as follows: The starting material (SM) is acylated to obtain intermediate 4; Intermediate 4 reacts with tert-butyl (4-aminophenyl)carbamate through a substitution reaction to obtain intermediate 5; Intermediate 5 removes the BOC protecting group to obtain intermediate 6; Intermediate 6 is acylated to obtain the target compound I-b.

[0060] In the following specific examples, unless otherwise specified, the materials and reagents used are obtained by regular purchase.

[0061] Example 1 Preparation of Compound I-1

[0062]

[0063] The specific reaction steps are as follows: (a) Dissolve 2,4-dichloropyrimidine (1 mmol) and 2-fluorobenzylamine (1.2 mmol) in ethanol (5 mL), and stir the reaction at room temperature for 5 h. After the reaction is completed, dilute with ethyl acetate (15 mL), wash twice with saturated sodium bicarbonate aqueous solution (10 mL), then wash with saturated brine (10 mL), add anhydrous sodium sulfate for drying for 30 min, filter and concentrate, and separate by column chromatography (the eluent is EA:PE = 10% - 30%) to obtain intermediate 1-1.

[0064] (b) Dissolve intermediate 1-1 (1 mmol) and tert-butyl (4-aminophenyl)carbamate (1.2 mmol) in n-butanol (5 mL). After complete dissolution by ultrasonic oscillation, add 1 drop of hydrochloric acid, and reflux the reaction under condensation at 80 °C in an oil bath for 8 h. After the reaction is completed, add ice water (15 mL), extract with ethyl acetate (30 mL × 2), combine the organic phases, wash with saturated brine (15 mL), add anhydrous sodium sulfate for drying, filter and concentrate, and separate by column chromatography (the eluent is PE:EA = 10% - 60%) to obtain intermediate 2-1.

[0065] (c) Dissolve intermediate 2-1 (1 mmol) in dichloromethane (4 mL), add trifluoroacetic acid (1.3 mmol), and react at room temperature for 3 h. After vacuum concentration, separate by column chromatography (the eluent is DCM:MeOH = 1% - 3%) to obtain intermediate 3-1.

[0066] (d) Dissolve intermediate 3-1 (1 mmol) in anhydrous dichloromethane (5 mL), slowly dropwise add benzoyl chloride (1.2 mmol) under ice bath conditions, then add triethylamine (2 mmol), stir and react at room temperature for 2 h, add 1 - 3 drops of hydrazine hydrate, and continue to react for 5 h. After the reaction is completed, rotary evaporate the reaction solution, then extract with ethyl acetate (20 mL × 3), wash twice with saturated aqueous sodium bicarbonate solution (20 mL), then wash with saturated brine (15 mL), add anhydrous sodium sulfate for drying for 30 min, filter and concentrate, and separate by column chromatography (the eluent is MeOH:DCM = 1% - 3%) to obtain compound I-1 in the form of a white solid, with a yield of 66.5% and a purity of 95%. MS(ESI) m / z(M+1) + : 414.1725.

[0067] 11H NMR (500 MHz, DMSO-d6) δ 10.07 (s, 1H), 8.94 (s, 1H), 7.98 – 7.94 (m, 2H), 7.84 (d, J = 5.8 Hz, 1H), 7.68 (s, 1H), 7.63 (d, J = 8.6 Hz, 1H), 7.57 (dt, J = 7.3, 2.7 Hz, 3H), 7.52 (dd, J = 8.2, 6.5 Hz, 2H), 7.39 (td, J = 7.7, 1.8 Hz, 1H), 7.31 (tdd, J = 7.5, 5.3, 1.8 Hz, 1H), 7.24 – 7.15 (m, 2H), 6.04 (d, J = 5.7 Hz, 1H), 4.67 – 4.56 (m, 2H). 13 13C NMR (126 MHz, DMSO) δ165.51, 162.98, 161.68, 160.16, 159.73, 137.73, 135.65, 132.74, 131.77, 129.64, 129.23, 128.78, 128.01, 126.90, 124.81, 121.23, 118.99, 115.62, 115.45, 37.74.

[0068] Example 2 Preparation of Compound I-2

[0069]

[0070] The preparation steps of Compound I-2 are basically the same as those in Example 1, except that in step (d), intermediate 3-1 and p-methoxybenzoyl chloride are used as raw materials for the reaction to obtain Compound I-2 in the form of a white solid, with a yield of 70.9% and a purity of 96%. MS(ESI) m / z(M+1) + : 444.1830.

[0071] 11H NMR (500 MHz, DMSO-d6) δ 13.29 (s, 1H), 10.31 (s, 1H), 8.61 (d, J = 3.2 Hz, 1H), 8.21 (d, J = 8.5 Hz, 1H), 7.98 – 7.89 (m, 2H), 7.81 (t, J = 7.9 Hz, 2H), 7.66 (d, J = 7.8 Hz, 1H), 7.57 (d, J = 16.4 Hz, 1H), 7.51 (d, J = 7.8 Hz, 1H), 7.47 (d, J = 8.0 Hz, 2H), 7.42 (q, J = 7.8 Hz, 2H), 7.28 (dd, J = 7.7, 4.9 Hz, 1H), 7.22 – 7.12 (m, 3H), 3.82 (s, 3H). 13 13C NMR (126 MHz, DMSO) δ 167.45, 165.92, 159.65, 155.36, 150.04, 140.76, 137.34, 136.55, 135.22, 133.98, 132.00, 130.05, 129.15, 126.83, 124.05, 123.13, 123.08, 122.19, 120.36, 120.33, 117.92, 113.41, 65.50, 55.81.

[0072] Example 3 Preparation of Compound I-3

[0073]

[0074] The preparation steps of Compound I-3 are basically the same as those in Example 1, except that in step (d), intermediate 3-1 and 3-methylbenzoyl chloride are used as raw materials for reaction to obtain Compound I-3 in the form of a white solid, with a yield of 71.2% and a purity of 96%. MS(ESI) m / z(M+1) + : 428.1881.

[0075] 1H NMR (500 MHz, DMSO-d6) δ 10.03 (d, J = 4.1 Hz, 1H), 8.92 (s, 1H), 7.85 (t, J = 4.8 Hz, 1H), 7.80 – 7.72 (m, 2H), 7.71 – 7.54 (m, 5H), 7.40 (dd, J = 7.3, 4.2 Hz, 3H), 7.31 (s, 1H), 7.24 – 7.13 (m, 2H), 6.04 (d, J = 5.8 Hz, 1H), 4.62 (s, 2H), 2.40 (d, J = 4.2 Hz, 3H). 13C NMR (126 MHz, DMSO) δ 165.62, 162.98, 161.67, 160.22, 159.73, 138.08, 137.71, 135.65, 132.77, 132.34, 129.59, 129.22, 129.16, 128.68, 128.51, 127.04, 125.16, 124.78, 121.19, 118.97, 115.62, 115.45, 21.43.

[0076] Example 4 Preparation of Compound I-4

[0077]

[0078] The preparation steps of Compound I-4 were basically the same as those in Example 1, except that in step (d), intermediate 3-1 and 2-chloromethylbenzoyl chloride were used as raw materials for the reaction to obtain Compound I-4 in the form of a white solid, with a yield of 71.2% and a purity of 97%. MS(ESI) m / z(M+1) + : 448.1335.

[0079] 1H NMR (500 MHz, DMSO-d6) δ 10.29 (s, 1H), 9.02 (s, 1H), 7.83 (d, J = 5.9 Hz, 2H), 7.60 (d, J = 8.5 Hz, 2H), 7.58 – 7.52 (m, 4H), 7.49 (td, J = 7.7, 1.9 Hz, 1H), 7.45 (dd, J = 7.4, 1.4 Hz, 1H), 7.41 – 7.36 (m, 1H), 7.31(d, J = 7.9 Hz, 1H), 7.23 – 7.13 (m, 2H), 6.05 (d, J = 5.9 Hz, 1H), 4.65 – 4.55 (m, 2H). 13C NMR (126 MHz, DMSO) δ 164.89, 162.98, 161.68, 159.73, 159.64, 137.69, 137.46, 132.88, 131.36, 130.45, 130.08, 129.67, 129.41, 129.23, 127.66, 126.86, 124.79, 120.33, 119.38, 115.63, 115.46, 39.53.

[0080] Example 5 Preparation of Compound I-5

[0081]

[0082] The specific reaction steps are as follows: (a) Replace the gas in the system of 2-chloro-4-aminopyrimidine (1 mmol), o-fluorobenzoic acid (1.2 mmol) and NaH (2.5 mmol) with a double-row tube, flush with nitrogen three times, and then dissolve in anhydrous DMF (5 mL) under ice bath conditions and stir at room temperature for 7 h. After the reaction is completed, quench with ice water, dilute with ethyl acetate, wash twice with saturated aqueous sodium bicarbonate solution (20 mL), then wash with saturated brine (15 mL), add anhydrous sodium sulfate and dry for 30 min, filter and concentrate, and separate by column chromatography (the eluent is EA:PE = 10% - 30%) to obtain intermediate 4-1.

[0083] (b)Dissolve intermediate 4-1 (1 mmol) and tert-butyl (4-aminophenyl)carbamate (1.2 mmol) in n-butanol (5 mL). After complete dissolution by ultrasonic oscillation, add 1 drop of hydrochloric acid, and reflux the reaction under a oil bath at 80 °C for 8 h. After the reaction is completed, add ice water (15 mL), extract with ethyl acetate (30 mL×2), combine the organic phases, wash with saturated brine (15 mL), add anhydrous sodium sulfate for drying, filter and concentrate, and separate by column chromatography (the eluent is EA:PE = 10%~60%) to obtain intermediate 5-1.

[0084] (c)Dissolve intermediate 5-1 (1 mmol) in dichloromethane (4 mL), add trifluoroacetic acid (1.3 mmol), and react at room temperature for 3 h. After the reaction is completed, concentrate under vacuum, and separate by column chromatography (the eluent is MeOH:DCM = 1%~3%) to obtain intermediate 6-1.

[0085] (d)Dissolve intermediate 6-1 (1 mmol) in anhydrous dichloromethane. Slowly add cyclohexanecarbonyl chloride (1.2 mmol) dropwise under ice bath conditions, then add triethylamine (2 mmol), stir and react at room temperature for 2 h, add 1 - 3 drops of hydrazine hydrate, and continue to react for 5 h. After the reaction is completed, rotary evaporate the reaction solution, then extract with ethyl acetate (20 ml×3), wash twice with saturated aqueous sodium bicarbonate solution (20 mL), then wash with saturated brine (15 ml), add anhydrous sodium sulfate for drying for 30 min, filter and concentrate, and separate by column chromatography (the eluent is MeOH:DCM = 1%~3%) to obtain white solid compound I-5 with a yield of 66.5% and a purity of 97%. MS(ESI) m / z(M+1) + : 434.1987.

[0086] 1H NMR (500 MHz, DMSO-d6) δ 10.85 (s, 1H), 9.79 (s, 1H), 9.38 (s,1H), 8.39 (d, J = 5.5 Hz, 1H), 7.71 (t, J = 9.8 Hz, 3H), 7.66 – 7.58 (m, 1H),7.51 (dd, J = 11.0, 6.4 Hz, 3H), 7.36 (dd, J = 13.6, 6.1 Hz, 2H), 1.76 (t, J= 15.7 Hz, 4H), 1.64 (s, 1H), 1.40 (q, J = 10.9, 9.3 Hz, 2H), 1.29 – 1.20 (m,3H). 13C NMR (126 MHz, DMSO) δ 174.37, 164.82, 160.12, 160.07, 158.62,158.22, 136.09, 134.08, 133.64, 130.62, 125.09, 125.06, 119.83, 119.47,116.75, 116.57, 101.21, 45.17, 29.67, 25.74.

[0087] Example 6 Preparation of Compound I-6

[0088]

[0089] The preparation steps of Compound I-6 are basically the same as those in Example 5, except that in step (a), 2-chloro-4-aminopyrimidine and o-chlorobenzoic acid are used as raw materials to react to obtain Intermediate 4-2; correspondingly, Intermediate 5-2 is obtained through step (b); Intermediate 6-2 is obtained through step (c); finally, white solid Compound I-6 is obtained through step (d), with a yield of 71.7% and a purity of 92%. MS(ESI) m / z(M+1) + : 450.1691.

[0090] 1H NMR (500 MHz, DMSO-d6) δ 11.04 (s, 1H), 9.58 (s, 1H), 9.32 (s, 1H), 8.33 (d, J = 5.5 Hz, 1H), 7.67 (d, J = 8.8 Hz, 2H), 7.57 – 7.37 (m, 7H), 2.21 (s, 1H), 1.74 – 1.50 (m, 6H), 1.33 (d, J = 15.2 Hz, 2H), 1.19 (d, J = 9.2 Hz, 2H). 13C NMR (126 MHz, DMSO) δ 174.41, 174.37, 167.09, 160.10, 158.29, 136.64, 136.20, 133.88, 131.93, 130.30, 130.08, 129.46, 127.71, 119.91, 119.46, 101.26, 45.30, 29.66, 25.88, 25.72.

[0091] Example 7 Preparation of Compound I-7

[0092]

[0093] The preparation steps of Compound I-7 were basically the same as those in Example 5, except that in step (a), 2-chloro-4-aminopyrimidine and 2-chloro-6-fluorobenzoic acid were used as raw materials to react to obtain Intermediate 4-3; correspondingly, Intermediate 5-3 was obtained through step (b); Intermediate 6-3 was obtained through step (c); finally, white solid Compound I-7 was obtained through step (d), with a yield of 74.2% and a purity of 93%. MS(ESI) m / z(M+1) + : 454.1804.

[0094] 1H NMR (500 MHz, DMSO) δ 11.35 (s, 1H), 9.64 (s, 1H), 9.41 (s, 1H), 8.42 (d, J = 5.6 Hz, 1H), 7.70 (d, J = 8.6 Hz, 2H), 7.57 (d, J = 7.3 Hz, 1H), 7.50 (t, J = 9.3 Hz, 3H), 7.46 (d, J = 8.1 Hz, 1H), 7.38 (t, J = 8.7 Hz, 1H), 2.29 (t, J = 11.6 Hz, 1H), 1.76 (t, J = 13.1 Hz, 4H), 1.64 (d, J = 12.3 Hz, 1H), 1.46 – 1.35 (m, 2H), 1.24 (dt, J = 30.6, 15.3 Hz, 3H). 13C NMR (126 MHz, DMSO) δ 174.34, 162.47, 160.37, 160.15, 158.17, 157.90, 136.06, 134.05, 132.61, 131.32, 126.04, 125.89, 119.89, 119.60, 115.21, 101.17, 45.30, 29.66, 25.89, 25.73.

[0095] Example 8 Preparation of Compound I-8

[0096]

[0097] The preparation steps of Compound I-8 were basically the same as those in Example 5, except that in step (a), 2-chloro-4-aminopyrimidine and 2,6-dichlorobenzoic acid were used as raw materials to react to obtain Intermediate 4-4; correspondingly, Intermediate 5-4 was obtained through step (b); Intermediate 6-4 was obtained through step (c); finally, white solid Compound I-8 was obtained through step (d), with a yield of 73.9% and a purity of 95%. MS(ESI) m / z(M+1) + : 484.1302.

[0098] 1H NMR (500 MHz, DMSO) δ 11.33 (d, J = 7.6 Hz, 1H), 9.64 (s, 1H), 9.38 (d, J = 15.3 Hz, 1H), 8.40 (dd, J = 10.3, 5.4 Hz, 1H), 7.70 (d, J = 8.6 Hz, 2H), 7.58 (d, J = 7.9 Hz, 2H), 7.53 (t, J = 4.1 Hz, 2H), 7.50 (t, J = 6.1 Hz, 2H), 2.29 (tt, J = 11.8, 3.5 Hz, 1H), 1.80 – 1.72 (m, 4H), 1.41 (td, J = 12.3, 3.0 Hz, 2H), 1.29 – 1.19 (m, 4H). 13C NMR (126 MHz, DMSO) δ 174.39, 164.35, 160.30, 160.16, 158.03, 136.08, 134.03, 132.12, 131.42, 128.69, 119.61, 101.24, 45.30, 42.43, 29.66, 29.51, 25.88, 25.73.

[0099] Example 9 Preparation of Compound I-9

[0100]

[0101] The preparation steps of Compound I-9 were basically the same as those in Example 5, except that in step (a), 2-chloro-4-aminopyrimidine and 2,6-dimethylbenzoic acid were used as raw materials to react to obtain Intermediate 4-5; correspondingly, Intermediate 5-5 was obtained through step (b); Intermediate 6-5 was obtained through step (c); finally, white solid Compound I-9 was obtained through step (d), with a yield of 70.5% and a purity of 95%. MS(ESI) m / z(M+1) + : 444.2394.

[0102] 1H NMR (500 MHz, DMSO) δ 11.03 (s, 1H), 9.59 (s, 1H), 9.35 (s, 1H), 8.38 (d, J = 5.5 Hz, 1H), 7.76 (d, J = 8.6 Hz, 2H), 7.60 (d, J = 5.6 Hz, 1H), 7.51 – 7.45 (m, 2H), 7.26 (dd, J = 9.6, 5.6 Hz, 1H), 7.13 (d, J = 7.6 Hz, 2H), 2.28 (s, 7H), 1.76 (t, J = 12.8 Hz, 4H), 1.64 (s, 1H), 1.40 (dd, J = 22.9, 10.9 Hz, 2H), 1.23 (dd, J = 25.2, 12.5 Hz, 3H). 13C NMR (126 MHz, DMSO) δ 174.25, 170.27, 160.13, 159.93, 158.38, 138.19, 136.28, 134.10, 133.83, 129.19, 127.77, 119.85, 119.36, 101.24, 45.30, 29.68, 25.90, 25.75, 19.35.

[0103] Example 10 Preparation of Compound I-10

[0104]

[0105] The preparation steps of Compound I-10 are basically the same as those in Example 1, except that in step (a), 2,4-dichloropyrimidine and 2-chlorobenzylamine are used as raw materials to react to obtain Intermediate 1-2; correspondingly, Intermediate 2-2 is obtained through step (b); Intermediate 3-2 is obtained through step (c); finally, in step (d), Intermediate 3-2 and cyclohexanecarbonyl chloride are used as raw materials to react to obtain Compound I-10 in the form of a white solid, with a yield of 66.5% and a purity of 94%. MS(ESI) m / z(M+1) + : 436.1899.

[0106] 1H NMR (500 MHz, DMSO-d6) δ 9.58 (s, 1H), 8.83 (s, 1H), 7.83 (d, J =5.8 Hz, 1H), 7.71 (s, 1H), 7.48 (s, 2H), 7.47 (d, J = 2.1 Hz, 1H), 7.36 (dd,J = 9.2, 2.1 Hz, 2H), 7.32 – 7.26 (m, 2H), 6.05 (s, 1H), 4.61 (s, 2H), 2.28(t, J = 11.6 Hz, 1H), 1.75 (t, J = 9.7 Hz, 4H), 1.64 (d, J = 11.8 Hz, 1H),1.47 – 1.35 (m, 2H), 1.31 – 1.13 (m, 4H). 13C NMR (126 MHz, DMSO) δ 174.16,162.97, 160.19, 155.62, 137.28, 136.96, 133.20, 132.50, 130.12, 129.57,128.87, 127.61, 119.79, 118.92, 45.23, 41.80, 29.68, 29.47, 25.91, 25.76.

[0107] Example 11 Preparation of Compound I-11

[0108]

[0109] The preparation steps of Compound I-11 are basically the same as those in Example 1, except that in step (a), 2,4-dichloropyrimidine and 2-bromobenzylamine are used as raw materials to react to obtain Intermediate 1-3; correspondingly, Intermediate 2-3 is obtained through step (b); Intermediate 3-3 is obtained through step (c); finally, in step (d), Intermediate 3-3 and cyclohexanecarbonyl chloride are used as raw materials to react to obtain Compound I-11 in the form of a white solid, with a yield of 71.6% and a purity of 95%. MS(ESI) m / z(M+1) + : 480.1393.

[0110] 1H NMR (500 MHz, DMSO-d6) δ 9.55 (s, 1H), 8.85 (s, 1H), 7.84 (d, J =5.8 Hz, 1H), 7.70 (dt, J = 8.0, 3.7 Hz, 1H), 7.64 (d, J = 7.7 Hz, 1H), 7.49(s, 1H), 7.38 (s, 1H), 7.36 – 7.33 (m, 3H), 7.32 (d, J = 5.7 Hz, 1H), 7.23 –7.19 (m, 1H), 6.05 (s, 1H), 4.57 (s, 2H), 2.28 (t, J = 11.5 Hz, 1H), 1.76 (s,3H), 1.64 (d, J = 12.0 Hz, 2H), 1.40 (d, J = 12.0 Hz, 2H), 1.26 – 1.21 (m,3H). 13C NMR (126 MHz, DMSO) δ 175.17, 174.15, 162.96, 160.15, 136.94,133.19, 132.81, 132.59, 129.20, 128.86, 128.16, 122.81, 122.51, 119.83,118.92, 45.24, 42.72, 29.68, 25.91, 25.76.

[0111] Example 12 Preparation of Compound I-12

[0112]

[0113] The preparation steps of Compound I-12 are basically the same as those in Example 1, except that in step (a), 2,4-dichloropyrimidine and 2-hydroxybenzylamine are used as raw materials to react to obtain Intermediate 1-4; correspondingly, Intermediate 2-4 is obtained through step (b); Intermediate 3-4 is obtained through step (c); finally, in step (d), Intermediate 3-4 and cyclohexanecarbonyl chloride are used as raw materials to react to obtain Compound I-12 in the form of a white solid, with a yield of 72.8% and a purity of 96%. MS(ESI) m / z(M+1) + : 418.2238.

[0114] 1 1H NMR (500 MHz, DMSO- d 6) δ 9.58 (d, J = 9.7 Hz, 2H), 8.89 (s, 1H), 7.78(d,J = 5.9 Hz, 1H), 7.74 – 7.65 (m, 1H), 7.59 (d, J = 8.6 Hz, 2H), 7.41 (d, J =8.6 Hz, 2H), 7.15 (d, J = 7.6 Hz, 1H), 7.06 (t, J = 7.7 Hz, 1H), 6.84 (d, J = 8.0Hz, 1H), 6.74 (t, J = 7.4 Hz, 1H), 6.01 (s, 1H), 4.47 (s, 1H), 4.22 (t, J = 6.5Hz, 1H), 2.28 (s, 1H), 1.76 (s, 2H), 1.67 – 1.61 (m, 2H), 1.38 (s, 1H), 1.23(s, 3H), 0.94 – 0.85 (m, 2H). 13 C NMR (126 MHz, DMSO) δ 174.18, 167.42, 163.11,159.68, 155.49, 136.79, 133.38, 131.96, 129.12, 128.23, 119.92, 119.38,115.39, 65.48, 45.24, 30.48, 29.68, 25.75, 19.11, 13.99.

[0115] Example 13 Preparation of Compound I-13

[0116]

[0117] The preparation steps of Compound I-13 are basically the same as those in Example 1, except that in step (a), 2,4-dichloropyrimidine and 2-methoxybenzylamine are used as raw materials to react to obtain Intermediate 1-5; correspondingly, Intermediate 2-5 is obtained through step (b); Intermediate 3-5 is obtained through step (c); finally, in step (d), Intermediate 3-5 and cyclohexanecarbonyl chloride are used as raw materials to react to obtain Compound I-13 in the form of a white solid, with a yield of 70.7% and a purity of 96%. MS(ESI) m / z(M+1) + : 432.2374.

[0118] 1H NMR (500 MHz, DMSO-d6) δ 9.56 (s, 1H), 8.80 (s, 1H), 7.79 (d, J = 5.8 Hz, 1H), 7.51 (s, 3H), 7.37 (d, J = 8.5 Hz, 2H), 7.22 (dd, J = 16.4, 8.0 Hz, 2H), 7.01 (d, J = 8.1 Hz, 1H), 6.89 (t, J = 7.4 Hz, 1H), 6.00 (s, 1H), 4.49 (s, 2H), 3.86 (s, 3H), 2.28 (t, J = 11.6 Hz, 1H), 1.86 (d, J = 36.9 Hz, 1H), 1.79 – 1.73 (m, 4H), 1.64 (s, 1H), 1.40 (d, J = 14.3 Hz, 2H), 1.23 (s, 2H). 13C NMR (126 MHz, DMSO) δ 174.33, 163.11, 159.58, 157.29, 155.49, 136.66, 133.44, 128.44, 127.96, 120.58, 119.79, 119.41, 115.46, 113.30, 110.94, 55.82, 47.81, 45.28, 29.67, 25.74, 14.00.

[0119] Example 14 Preparation of Compound I-14

[0120]

[0121] The preparation steps of Compound I-14 are basically the same as those in Example 1, except that in step (a), 2,4-dichloropyrimidine and 2-methylbenzylamine are used as raw materials to react to obtain Intermediate 1-6; correspondingly, Intermediate 2-6 is obtained through step (b); Intermediate 3-6 is obtained through step (c); finally, in step (d), Intermediate 3-6 and cyclohexanecarbonyl chloride are used as raw materials to react to obtain Compound I-14 in the form of a white solid, with a yield of 75.0% and a purity of 94%. MS(ESI) m / z(M+1) + : 416.2445.

[0122] 1H NMR (500 MHz, DMSO-d6) δ 9.64 (s, 1H), 8.88 (s, 1H), 7.85 (d, J = 5.7 Hz, 1H), 7.59 (s, 3H), 7.43 (d, J = 8.6 Hz, 2H), 7.34 – 7.15 (m, 4H), 6.06 (s, 1H), 4.56 (s, 2H), 2.37 (s, 4H), 1.81 (d, J = 11.7 Hz, 4H), 1.67 (d, J = 10.0 Hz, 2H), 1.50 – 1.36 (m, 3H), 1.31 (d, J = 3.0 Hz, 1H). 13C NMR (126 MHz, DMSO) δ 174.23, 167.46, 162.99, 160.19, 137.06, 135.97, 133.16, 130.39, 127.17, 126.22, 119.80, 119.06, 65.50, 45.27, 42.73, 29.68, 29.59, 25.91, 25.75, 19.14, 13.99.

[0123] Example 15 Preparation of Compound I-15

[0124]

[0125] The preparation steps of Compound I-15 were basically the same as those in Example 1, except that in step (a), 2,4-dichloropyrimidine and 2-(trifluoromethyl)benzylamine were used as raw materials to react to obtain Intermediate 1-7; correspondingly, Intermediate 2-7 was obtained through step (b); Intermediate 3-7 was obtained through step (c); finally, in step (d), Intermediate 3-7 and cyclohexanecarbonyl chloride were used as raw materials to react to obtain Compound I-15 in the form of a white solid, with a yield of 74.2% and a purity of 94%. MS(ESI) m / z(M+1) + : 470.2162.

[0126] 1H NMR (500 MHz, DMSO-d6) δ 9.55 (s, 1H), 8.87 (s, 1H), 7.85 (d, J = 5.8 Hz, 1H), 7.76 (d, J = 7.8 Hz, 1H), 7.72 (s, 1H), 7.64 (t, J = 7.6 Hz, 1H), 7.53 (d, J = 7.8 Hz, 1H), 7.47 (t, J = 7.5 Hz, 3H), 7.33 (d, J = 7.5 Hz, 2H), 6.06 (s, 1H), 4.76 (s, 2H), 2.27 (t, J = 11.6 Hz, 1H), 1.75 (t, J = 10.4 Hz, 4H), 1.64 (d, J = 11.6 Hz, 1H), 1.44 – 1.35 (m, 2H), 1.24 (d, J = 6.0 Hz, 2H), 0.93 – 0.81 (m, 1H). 13C NMR (126 MHz, DMSO) δ 174.16, 163.05, 160.10, 142.63, 138.43, 136.81, 133.11, 130.96, 128.69, 127.70, 127.16, 126.74, 126.51, 126.29, 119.73, 118.93, 48.77, 45.24, 29.67, 25.91, 25.75.

[0127] Example 16 Preparation of Compound I-16

[0128]

[0129] The preparation steps of Compound I-16 are basically the same as those in Example 1, except that in step (a), 2,4-dichloropyrimidine and 2-pyridylmethylamine are used as raw materials to react to obtain Intermediate 1-8; correspondingly, Intermediate 2-8 is obtained through step (b); Intermediate 3-8 is obtained through step (c); finally, in step (d), Intermediate 3-8 and cyclohexanecarbonyl chloride are used as raw materials to react to obtain Compound I-16 in the form of a white solid, with a yield of 67.7% and a purity of 90%. MS(ESI) m / z(M+1) + : 403.2241.

[0130] 1H NMR (500 MHz, DMSO) δ 9.56 (s, 1H), 8.83 (s, 1H), 8.54 (d, J = 4.9Hz, 1H), 7.82 (d, J = 5.8 Hz, 1H), 7.74 (d, J = 1.8 Hz, 2H), 7.49 (s, 2H),7.37 (d, J = 8.5 Hz, 2H), 7.32 (d, J = 7.9 Hz, 1H), 7.28 – 7.23 (m, 1H), 6.03(s, 1H), 4.62 (d, J = 2.2 Hz, 2H), 2.28 (s, 1H), 1.76 (t, J = 13.7 Hz, 4H),1.65 (d, J = 12.0 Hz, 1H), 1.45 – 1.34 (m, 2H), 1.24 (dt, J = 28.4, 14.2 Hz,3H). 13C NMR (126 MHz, DMSO) δ 175.18, 174.17, 163.03, 160.14, 159.67,155.56, 149.36, 137.13, 136.93, 133.20, 122.47, 121.30, 119.82, 119.11,45.93, 45.27, 29.69, 25.91, 25.76.

[0131] Example 17 Preparation of Compound I-17

[0132]

[0133] The preparation steps of Compound I-17 are basically the same as those in Example 1, except that in step (a), 2,4-dichloropyrimidine and 3-methoxybenzylamine are used as raw materials to react to obtain Intermediate 1-9; correspondingly, Intermediate 2-9 is obtained through step (b); Intermediate 3-9 is obtained through step (c); finally, in step (d), Intermediate 3-9 and cyclohexanecarbonyl chloride are used as raw materials to react to obtain Compound I-17 in the form of a white solid, with a yield of 65.7% and a purity of 97%. MS(ESI) m / z(M+1) + : 432.2374.

[0134] 1H NMR (500 MHz, DMSO) δ 9.58 (s, 1H), 8.82 (s, 1H), 7.80 (d, J = 5.6Hz, 1H), 7.62 (d, J = 21.2 Hz, 1H), 7.57 (d, J = 7.4 Hz, 2H), 7.40 (d, J =8.8 Hz, 2H), 7.24 (t, J = 8.0 Hz, 1H), 6.91 (d, J = 7.5 Hz, 2H), 6.82 – 6.77(m, 1H), 5.97 (s, 1H), 4.50 (s, 2H), 3.71 (s, 3H), 2.28 (ddd, J = 11.7, 7.6,3.2 Hz, 1H), 1.76 (t, J = 12.5 Hz, 4H), 1.65 (d, J = 11.8 Hz, 1H), 1.45 –1.35 (m, 2H), 1.31 – 1.19 (m, 3H). 13C NMR (126 MHz, DMSO) δ 174.24, 162.09,159.97, 159.87, 157.90, 156.82, 136.60, 133.37, 127.79, 119.73, 118.98,116.46, 112.49, 112.30, 97.32, 45.25, 29.67, 25.90, 25.75.

[0135] Example 18 Preparation of Compound I-18

[0136]

[0137] The preparation steps of Compound I-18 were basically the same as those in Example 1, except that in step (a), 2,4-dichloropyrimidine and 3-hydroxybenzylamine were used as raw materials to react to obtain Intermediate 1-10; correspondingly, Intermediate 2-10 was obtained through step (b); Intermediate 3-10 was obtained through step (c); finally, in step (d), Intermediate 3-10 and cyclohexanecarbonyl chloride were used as raw materials to react to obtain Compound I-18 in the form of a white solid, with a yield of 64.3% and a purity of 94%. MS(ESI) m / z(M+1) + : 418.2238.

[0138] 1H NMR (500 MHz, DMSO) δ 9.57 (s, 1H), 9.31 (s, 1H), 8.84 (s, 1H), 7.80 (d, J = 5.4 Hz, 1H), 7.63 (dd, J = 45.1, 6.5 Hz, 3H), 7.40 (d, J = 8.7 Hz, 2H), 7.12 (t, J = 7.8 Hz, 1H), 6.77 – 6.71 (m, 2H), 6.63 (dd, J = 8.0, 1.5 Hz, 1H), 5.97 (s, 1H), 4.47 (s, 2H), 3.35 (s, 2H), 2.32 – 2.24 (m, 1H), 1.76 (t, J = 13.2 Hz, 4H), 1.64 (d, J = 11.7 Hz, 1H), 1.46 – 1.34 (m, 2H), 1.23 (s, J = 13.1 Hz, 3H). 13C NMR (126 MHz, DMSO) δ 175.00, 174.16, 162.97, 160.14, 157.91, 155.05, 141.82, 137.03, 133.17, 129.68, 119.85, 119.12, 118.14, 114.21, 114.12, 97.93, 45.25, 29.69, 25.91, 25.76.

[0139] Example 19 Preparation of Compound I-19

[0140]

[0141] The preparation steps of Compound I-19 are basically the same as those in Example 1, except that in step (a), 2,4-dichloropyrimidine and 4-methoxybenzylamine are used as raw materials to react to obtain Intermediate 1-11; correspondingly, Intermediate 2-11 is obtained through step (b); Intermediate 3-11 is obtained through step (c); finally, in step (d), Intermediate 3-11 and cyclohexanecarbonyl chloride are used as raw materials to react to obtain Compound I-19 in the form of a white solid, with a yield of 66.6% and a purity of 98%. MS(ESI) m / z(M+1) + : 432.2734.

[0142] 1 1H NMR (500 MHz, DMSO) δ 9.58 (s, 1H), 8.86 (s, 1H), 7.78 (d, J= 5.5 Hz, 1H), 7.59 (d, J = 8.1 Hz, 3H), 7.41 (d, J = 8.7 Hz, 2H), 7.26 (d, J = 8.5 Hz, 2H), 6.91 – 6.86 (m, 2H), 5.94 (d, J = 4.1 Hz, 1H), 4.45 (s, 2H), 2.32 – 2.25 (m, 1H), 2.04 (tt, J = 11.7, 3.4 Hz, 2H), 1.75 (dd, J = 18.5, 7.9 Hz, 4H), 1.69 (dt, J = 20.2, 6.9 Hz, 4H), 1.23 (t, J = 3.9 Hz, 3H). 13 C NMR (126 MHz, DMSO) δ 175.16, 174.15, 160.18, 158.65, 137.06, 133.17, 128.89, 119.81, 119.12, 114.17, 55.50, 45.25, 42.72, 29.68, 29.60, 25.89, 25.74.

[0143] Example 20 Preparation of Compound I-20

[0144]

[0145] The preparation steps of Compound I-20 were basically the same as those in Example 1, except that in step (a), 2,4-dichloropyrimidine and 4-hydroxybenzylamine were used as raw materials to react to obtain Intermediate 1-12; correspondingly, Intermediate 2-12 was obtained through step (b); Intermediate 3-12 was obtained through step (c); finally, in step (d), Intermediate 3-12 and cyclohexanecarbonyl chloride were used as raw materials to react to obtain Compound I-20 in the form of a white solid, with a yield of 64.5% and a purity of 95%. MS(ESI) m / z(M+1) + : 418.2238.

[0146] 1 H NMR (500 MHz, DMSO) δ 9.57 (s, 1H), 9.31 (s, 1H), 8.84 (s, 1H), 7.80 (d, J = 5.4 Hz, 1H), 7.63 (dd,J = 45.1, 6.5 Hz, 3H), 7.40 (d, J = 8.7 Hz,2H), 7.12 (t, J = 7.8 Hz, 1H), 6.77 – 6.71 (m, 2H), 6.63 (dd, J = 8.0, 1.5 Hz,1H), 5.97 (s, 1H), 4.47 (s, 2H), 2.32 – 2.24 (m, 1H), 1.76 (t, J = 13.2 Hz,4H), 1.64 (d, J = 11.7 Hz, 1H), 1.46 – 1.34 (m, 2H), 1.23 (s, 3H). 13 C NMR (126MHz, DMSO) δ 174.19, 172.52, 162.87, 159.97, 156.73, 136.95, 133.29, 130.30,128.97, 119.83, 119.26, 115.52, 45.26, 29.69, 26.81, 25.92, 25.76, 21.53.

[0147] Example 21 Preparation of Compound I-21

[0148]

[0149] The preparation steps of Compound I-21 are basically the same as those in Example 1, except that in step (a), 2,4-dichloropyrimidine and 4-(trifluoromethyl)benzylamine are used as raw materials to react to obtain Intermediate 1-13; correspondingly, Intermediate 2-13 is obtained through step (b); Intermediate 3-13 is obtained through step (c); finally, in step (d), Intermediate 3-13 and cyclohexanecarbonyl chloride are used as raw materials to react to obtain Compound I-21 in the form of a white solid, with a yield of 74.7% and a purity of 95%. MS(ESI) m / z(M+1) + : 470.2162.

[0150] 1H NMR (500 MHz, DMSO) δ 9.57 (s, 1H), 8.82 (d, J = 4.3 Hz, 1H), 7.82 (d, J = 5.7 Hz, 1H), 7.76 (s, 1H), 7.69 (d, J = 8.0 Hz, 2H), 7.53 (dd, J = 18.5, 8.0 Hz, 4H), 7.38 (d, J = 8.5 Hz, 2H), 5.99 (s, 1H), 4.62 (s, 2H), 2.28 (t, J = 11.6 Hz, 1H), 1.77 (d, J = 11.3 Hz, 4H), 1.65 (d, J = 11.8 Hz, 1H), 1.47 – 1.35 (m, 2H), 1.31 – 1.16 (m, 3H). 13C NMR (126 MHz, DMSO) δ 174.30, 174.25, 162.94, 160.23, 145.48, 136.92, 133.23, 128.14, 128.01, 127.76, 125.92, 125.60, 123.76, 119.88, 119.23, 45.26, 29.67, 25.90, 25.73.

[0151] The following studies on the activity and selectivity of the compounds prepared in Examples 1 - 21 were carried out:

[0152] 1. Inhibitory effect of the compounds prepared in Examples 1 - 21 on BaF3 cells

[0153] 1) BaF3 cells with a cell density of approximately 4×10 4 cells / mL were evenly mixed with the test compounds at different concentrations, and 100 μL was taken and placed into 96-well plates respectively;

[0154] 2) The 96-well plates were placed in a cell culture incubator and cultured at 37 °C for 72 h;

[0155] 3) The surviving BaF3 cells were tested using the CellTiter-Glo kit; during the CellTiter-Glo test, the fluorescence data was read using a multi-label reader (Envision, PerkinElmer, USA); all test data was referenced with DMSO;

[0156] 4) The IC 50 was calculated using data processing software. The experimental results are shown in Table 1.

[0157] 2. Inhibitory effects of the compounds prepared in Examples 1-21 on BaF3-TYK2 cells

[0158] The in vitro inhibitory activity of the compounds on BaF3-TYK2 cells was detected using an ADP-Glo kit (Promega). The reaction system included: 4.95 μL of protein, 0.55 μL of serially diluted compounds, 5.5 μL of substrate and peptide (4:1 Gly, Tyr, 0.2), and 100 μM of ATP. After adding ATP, the reaction was carried out at 37 °C for 1 h and then cooled at room temperature for 5 min. 5 μL of the reaction solution was added to a 384-well plate, and then 5 μL of ADP-Glo reagent was added to terminate the reaction. Finally, 10 μL of kinase detection reagent was added and incubated for 30 min. The experimental results were read using a Perkin-Elmer Envision to measure the fluorescence value and calculate the IC 50 value. The experimental results are shown in Table 1.

[0159] Table 1: Experimental results of the inhibitory effects of the compounds prepared in Examples 1-21 on BaF3 and BaF3-TYK2 cells

[0160]

[0161] In this invention, cerdulatinib was used as a positive control to conduct safety tests on the compounds prepared in Examples 1-21 for BaF3 and BaF3-TYK2 cells. The research results showed that most of the compounds had good safety for BaF3 cells, and at a concentration of 1 μM, the compounds significantly inhibited the growth of BaF3-TYK2 cells.

[0162] 3. Experiment on the effect of Compound I-13 on the JAK-STAT signaling pathway

[0163] TYK2 is an important member of the JAK-STAT signaling pathway, mainly mediating type I interferon, IL-12, and IL-23 signals and regulating immune and inflammatory responses. To identify whether I-13 can inhibit the activation of the JAK-STAT signaling pathway by inhibiting the protein activity of TYK2, the expression of STAT3 phosphorylation in this signaling pathway was detected by Western blotting.

[0164] 3.1 Experimental method

[0165] RAW 264.7 cells in logarithmic growth phase were prepared into a cell suspension of 8×10 4The suspension at [X] cells / mL was inoculated into a 6-well plate at 1 mL / well. A total of 6 groups were set up, namely the Control group, the IFN-α group, the I-13 + IFN-α groups (with I-13 concentrations of 2.5 μM, 5 μM, and 10 μM respectively), and the Cerdulatinib group. After culturing in an incubator for 24 h, except for the Control group, 5 μL of 20 μg / mL IFN-α was added to stimulate the cells in each of the other groups. At the same time, except for the Control group and the IFN-α group, 1 mL of compound I-13 solution with concentrations of 2.5, 5, and 10 μM was added to each well of the dosing groups, and 1 mL of Cerdulatinib solution with a concentration of 1 μM was added to the Cerdulatinib group. After culturing in the incubator for 24 h, proteins were extracted for Western Blot experiments to detect the phosphorylation of STAT3 protein.

[0166] 3.2 Experimental Results

[0167] The results of the Western Blot experiment showed (see Figure 1 ) that compound I-13 began to exert an inhibitory effect on the JAK-STAT signaling pathway at a concentration of 2.5 μM. The inhibitory effect was the strongest at a concentration of 10 μM, indicating that within the safe concentration range, the inhibitory effect of compound I-13 on the JAK-STAT3 signaling pathway gradually increased with the increase in concentration. In summary, compound I-13 can inhibit the expression of p-STAT3 protein, thereby blocking the conduction of JAK-STAT pathway signals.

[0168] The above results indicate that compound I-13 of the present invention has good TYK2 inhibitory activity. At the same time, through experimental verification by the inventors, the 21 compounds prepared in Examples 1 to 21 have good TYK2 inhibitory activity and can all be used as TYK2 selective inhibitors for the treatment of autoimmune diseases, especially suitable for the treatment of rheumatoid arthritis, ulcerative colitis, psoriasis, skin inflammation, and other inflammatory and autoimmune diseases.

[0169] The above specific embodiments have described the implementation of the present invention in detail. However, the present invention is not limited to the specific details in the above embodiments. Within the scope of the claims and technical concept of the present invention, various simple modifications and changes can be made to the technical solutions of the present invention, and these simple variations all fall within the protection scope of the present invention.

Claims

1. A selective TYK2 inhibitor compound, characterized in that, It has the structure shown below: ； Wherein, X is -NH-(C=O)- or -NH-(CH2)-; R1 is a heteroaryl or a substituted aryl, and the substituents of the aryl include but are not limited to halogen, trifluoromethyl, methoxy, hydroxyl or methyl; R2 is an alkane, an aryl or a substituted aryl, and the substituents of the aryl include but are not limited to halogen, trifluoromethyl, methoxy, hydroxyl or methyl.

2. The selective TYK2 inhibitor compound according to claim 1, wherein, The compound has the structure shown by the following formula: ； Wherein, R1 is pyridine or a substituted aryl, and the substituents of the aryl are halogen, trifluoromethyl, methoxy, hydroxyl or methyl; R2 is an alkane, an aryl or a substituted aryl, and the substituents of the aryl are halogen, trifluoromethyl, methoxy, hydroxyl or methyl.

3. An alternative TYK2 inhibitor compound according to claim 1, characterized in that, The compound has the structure shown by the following formula: ； Wherein, R1 is 2-fluorophenyl, 2-chlorophenyl, 2-fluoro-6-chlorophenyl or 2,6-dimethylphenyl; R2 is an alkane, an aryl or a substituted aryl, and the substituents of the aryl are halogen, trifluoromethyl, methoxy, hydroxyl or methyl.

4. A selective TYK2 inhibitor compound according to claim 1, wherein The R2 is selected from one of the following structures: 。 5. A selective TYK2 inhibitor compound according to claim 1, wherein The compound has one of the following structures: ； 。 6. The preparation method of a selective TYK2 inhibitor compound according to claim 1, characterized in that, When X is -NH-(CH2)-, the compound is prepared by the following reaction route: ； When X is -NH-(C=O)-, the compound is prepared by the following reaction route: 。 7. Use of the compound according to any one of claims 1 to 5 or a pharmaceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof in the preparation of a drug for a TYK2 inhibitor or a JAK inhibitor.

8. The application according to claim 7, characterized in that, The JAK includes JAK1, JAK2, JAK3.

9. Use of the compound according to any one of claims 1 to 5 or a pharmaceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof in the preparation of a medicament for preventing or treating autoimmune diseases, characterized in that, The autoimmune diseases include but are not limited to rheumatoid arthritis, psoriasis, Crohn's disease, inflammatory bowel disease, ulcerative colitis or autoimmune-related skin diseases.

10. A pharmaceutical composition, characterized in that, It includes the compound according to any one of claims 1 to 5 or a pharmaceutically acceptable salt, solvate, ester, acid, metabolite or prodrug thereof, and one or more pharmaceutically acceptable carriers, diluents or excipients.

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