CN110330484A - Replace class phenyl pyrimidine derivative as jak kinase inhibitor or its pharmaceutical salt, Preparation method and use - Google Patents

Abstract

The present invention discloses a substituted phenylpyrimidine derivative having the general formula (I) as a JAK kinase inhibitor or a pharmaceutically acceptable salt thereof, a preparation method and an application thereof. The compound has excellent JAK inhibitory effect, and it is used as a drug for preventing, treating or improving autoimmune diseases, Sjogren's syndrome, Behcet's disease, multiple sclerosis, systemic lupus erythematosus and the like. The compounds of the present invention exhibit highly active JAK-3 inhibitory action IC ₅₀ up to 1.7nM, and the present invention has a simple synthetic route and strong practicability.

Description

Substituted phenylpyrimidine derivatives as JAK kinase inhibitors or pharmaceutically acceptable salts thereof, Preparation method and use

technical field

The present invention relates to kinase inhibitors, preparation methods and applications, in particular to substituted phenylpyrimidine derivatives as JAK kinase inhibitors or their pharmaceutically acceptable salts, preparation methods and uses.

Background technique

There are four isoforms of JAK kinase (janus kinase): JAK-1, JAK-2, JAK-3 and TYK-2, which interact with downstream effectors, signal transducers and activators of transcription proteins (STATs) ) formed an important cytokine signaling pathway, the JAK-STAT pathway (Science, 1994, 264: 1415-1421). The discovery of this pathway has greatly improved researchers' understanding of gene regulation. Studies have found that the JAK-STAT pathway can be activated by a variety of cytokines, growth factors and receptors, and is involved in cell proliferation, differentiation, apoptosis, angiogenesis, and immune regulation. Process (World J Gastroenterol, 2007, 13:6478-6491). JAK kinases mediate the signaling of most cytokines in cells, such as interleukins (IL), interferons (IFN), erythropoietin (EPO), granulocyte and macrophage colony stimulating factor (GMCSF), Growth hormone (GH), prolactin (PRL), thrombopoietin (TPO), platelet-derived factor (PDGF) and epidermal growth factor (EGF), etc., and different receptors can activate different subtypes of JAK kinases.

Among them, JAK-3 regulates the regulation by binding to γ chain (γc) in type I cytokine receptor complexes such as IL-2, IL-4, IL-7, IL-9, IL-15 and IL-21. Cell Signaling. When JAK-3 deficiency or γc mutation can lead to severe combined immunodeficiency (SCID), manifested as decreased T cells and natural killer cells (NK) cells, loss of B cell function and other immune restriction symptom. These cytokines play an important role in immune responses when present in appropriate amounts. However, their overproduction is believed to be associated with many autoimmune diseases such as psoriasis, rheumatoid arthritis, inflammatory bowel disease, Sjogren's syndrome, Behcet's disease, multiple sclerosis, systemic erythema Lupus et al. (Journal of Allergy and Clinical Tmmurmlogy 127, 3, 701-721.e70 (2011), Cytokine & Growth Factor Reviews 19, 41-52 (2008), Invest Ophthalmol Vis Sci. 2008 Jul; 49(7): 3058-3064, Ann Rheum Dis 2010 Jul;69(7):1325-1328). Furthermore, JAK-1, JAK-2 and TYK-2 are widely present in various tissues and cells in the body, and JAK3 mainly exists in bone marrow cells, thymocytes, NK cells and activated B lymphocytes and T lymphocytes. -3 is recognized as a relatively safe and effective anti-autoimmune target in the JAK-STAT pathway.

How to inhibit the binding of ATP to the ATP binding site on JAK, block the hydrolysis of ATP, interfere with the phosphorylation of JAKs, thereby prevent the activation of JAKs, cut off their signal transmission to STATs, and prevent them from regulating the expression of genes in the nucleus, thereby preventing Disrupting the JAK-STAT signaling pathway is the focus of research.

SUMMARY OF THE INVENTION

Object of the invention: The object of the present invention is to provide substituted phenylpyrimidine derivatives as JAK kinase inhibitors or their pharmaceutically acceptable salts.

Another object of the present invention is to provide a method for preparing the substituted phenylpyrimidine derivative as a JAK kinase inhibitor or a pharmaceutically acceptable salt thereof.

The last object of the present invention is to provide the use of the substituted phenylpyrimidine-like derivatives as JAK kinase inhibitors or pharmaceutically acceptable salts thereof.

Technical solution: The present invention provides a substituted phenylpyrimidine derivative having the general formula (I) as a JAK kinase inhibitor or a pharmaceutically acceptable salt thereof,

in,

X is a nitrogen atom or -CH-;

Y is a nitrogen atom or -CH-;

Z is Substituted or unsubstituted C _3-8 heterocyclic alkyl;

W is a substituted 5-membered heteroaryl;

^R1 is

in,

V is a hydrogen atom, halogen or cyano;

R ⁴ , R ⁵ , R ⁶ and R ⁷ are respectively a hydrogen atom, a substituted or unsubstituted C _1-6 hydrocarbyl group, and a substituted or unsubstituted C _1-6 heterohydrocarbyl group;

R ² is a hydrogen atom, a halogen atom, a substituted or unsubstituted 4, 5, 6, 7- or 8-membered heterocycle;

R ³ is substituted C _1-6 hydrocarbyl, substituted or unsubstituted C _1-6 heterohydrocarbyl.

Further, the

Z is

Further, the R ² is a hydrogen atom, a fluorine atom, a substituted or unsubstituted 4, 5 or 6-membered nitrogen-containing heterocyclic ring. The W is a nitrogen-containing five-membered heteroaromatic ring. The R ³ is a methyl group or a substituted C _1-6 alkyl group, and the substituent of the C _1-6 alkyl group is a halogen atom, a hydroxyl group, a C _1-6 alkoxy group, an aliphatic amino group, and an amide group.

Further, a substituted phenylpyrimidine derivative having the general formula (I) as a JAK kinase inhibitor or a pharmaceutically acceptable salt thereof,

N-(3-(6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)acrylamide,

N-(3-(6-((1-Methyl-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)acrylamide,

N-(4-(6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)acrylamide,

N-(3-(2-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)acrylamide,

N-(4-(2-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)acrylamide,

N-(3-(6-(4-morpholinylamino)pyrimidin-4-yl)phenyl)acrylamide,

N-(4-(6-(4-morpholinylamino)pyrimidin-4-yl)phenyl)acrylamide,

N-(3-(2-(4-morpholinylamino)pyrimidin-4-yl)phenyl)acrylamide,

N-(4-(2-(4-morpholinylamino)pyrimidin-4-yl)phenyl)acrylamide,

N-(3-(6-((1-(2-N,N-dimethylethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)acrylamide,

N-(3-(6-((1-(N-methylacetamide)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)acrylamide,

N-(3-(6-((1-(2-hydroxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)acrylamide,

N-(3-(6-((1-Methyl-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)-2-butenamide,

N-(3-(6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)-2-butenamide,

N-(4-(6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)-2-butenamide,

N-(4-(2-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)-2-butenamide,

N-(3-(6-(4-morpholinylamino)pyrimidin-4-yl)phenyl)-2-butenamide,

N-(3-(2-(4-morpholinylamino)pyrimidin-4-yl)phenyl)-2-butenamide,

N-(4-(6-(4-morpholinylamino)pyrimidin-4-yl)phenyl)-2-butenamide,

N-(4-(2-(4-morpholinylamino)pyrimidin-4-yl)phenyl)-2-butenamide,

N-(3-(6-((1-(2-N,N-dimethylethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)-2-butane enamide,

N-(3-(6-((1-(N-methylacetamide)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)-2-butenamide,

N-(3-(6-((1-1-(2-hydroxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)-2-butenamide,

2-cyano-N-(3-(6-((1-methyl-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)acetamide,

2-cyano-N-(3-(6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)acetamide,

2-cyano-N-(4-(6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)acetamide,

2-cyano-N-(4-(2-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)acetamide,

2-cyano-N-(3-(((6-(4-morpholinoanilino)pyrimidin-4-yl)amino)methyl)phenyl)acetamide,

2-cyano-N-(3-(((2-(4-morpholinylamino)pyrimidin-4-yl)amino)methyl)phenyl)acetamide,

2-cyano-N-(4-(((6-(4-morpholinoanilino)pyrimidin-4-yl)amino)methyl)phenyl)acetamide,

2-cyano-N-(4-(((2-(4-morpholinoanilino)pyrimidin-4-yl)amino)methyl)phenyl)acetamide,

N-[2-oxo-2-((3-(6-(1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)-phenyl -amino)ethyl]acrylamide,

N-[2-oxo-2-((3-(6-(1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)-phenyl -amino)ethyl]butenamide,

N-[2-oxo-2-((3-(6-(4-morpholinoanilino)amino)pyrimidin-4-yl)-phenyl-amino)ethyl]acrylamide,

N-[2-oxo-2-((3-(6-(4-morpholinoanilino)amino)pyrimidin-4-yl)-phenyl-amino)ethyl]butenamide, 2-cyano -N-[2-oxo-2-((3-(6-(1-methyl-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)-phenyl-amino)ethyl] acetamide,

2-cyano-N-[2-oxo-2-((3-(6-(4-morpholinylamino)amino)pyrimidin-4-yl)-phenyl-amino)ethyl]acetamide,

2-cyano-N-[2-oxo-2-((3-(6-(1-(N-methylacetamide)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl )-phenyl-amino)ethyl]acetamide,

2-cyano-N-(4-(6-((1-1-(2-hydroxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)acetamide.

A pharmaceutical composition containing a therapeutically effective amount of one or more substituted phenylpyrimidine derivatives of general formula (I) as described in any one of claims 1-6 as JAK kinase inhibitors or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or adjuvant.

The described preparation method of the substituted phenylpyrimidine derivative with general formula (I) as a JAK kinase inhibitor or a pharmaceutically acceptable salt thereof comprises the following steps:

in,

Step 1: Compound III is converted into compound IV under the conditions of tetrakistriphenylphosphine palladium and basic potassium acetate with double pinacol boronate;

Step 2: add 2,4-chloro-pyrimidine or 4,6-chloro-pyrimidine, tetrakistriphenylphosphine palladium and potassium carbonate aqueous solution to the reaction solution of step 1 to carry out suziki reaction to obtain compound V;

Step 3: Compound V is reacted with a substituted amine to obtain Compound VI;

Step 4: the reduction reaction of compound VI, into compound VII;

Step 5: Compound VII is subjected to an amide-forming reaction with a R ¹ substitution reagent to obtain the compound of the present invention.

The substituted phenylpyrimidine derivatives with the general formula (I) are used as JAK kinase inhibitors or their pharmaceutically acceptable salts in the preparation and treatment of autoimmune diseases, Sjogren's syndrome, Behcet's disease, multiple Use in the drug of sexual sclerosis and systemic lupus erythematosus.

Further, the autoimmune disease is psoriasis, rheumatoid arthritis or inflammatory bowel disease.

The highly active and selective JAK inhibitor or a pharmaceutically acceptable salt thereof developed by the present invention competitively inhibits the binding of ATP to the ATP binding site on JAK, blocks the hydrolysis of ATP, interferes with the phosphorylation process of JAKs, thereby preventing JAKs The activation of JAK-STATs cuts off its signal transmission to STATs, resulting in its inability to regulate the expression of genes in the nucleus, thereby achieving the purpose of blocking the JAK-STAT signaling pathway.

Beneficial effect: The compound of the present invention has excellent JAK-1 and/or JAK-3 inhibitory effect, which is used for the prevention, treatment or improvement of autoimmune diseases (eg, psoriasis, rheumatoid arthritis, inflammatory bowel disease (eg, Crohn's disease, ulcerative colitis, etc.), Sjogren's syndrome, Behcet's disease, multiple sclerosis, systemic lupus erythematosus, etc.), etc., the compound of Example 5 showed high Active JAK-3 inhibitory effect (IC ₅₀ =1.7nm), moderate selective activity (JAK-1/JAK-3=586, JAK-2/JAK-3=60), the synthetic route of the present invention is simple and practical powerful.

Detailed ways

In the following examples, "room temperature" refers to about 10°C to about 35°C. The ratio indicated by the mixed solvent is the volume mixing ratio unless otherwise specified. Unless otherwise stated, % refers to % by weight.

In silica gel column chromatography, basic silica refers to silica bound with aminopropylsilane. In high performance liquid chromatography (HP1C), C18 refers to silica bound using octadecyl groups. Ratios of elution solvents are volume mixing ratios unless otherwise specified.

In the following examples and experimental examples, the following abbreviations are used.

THF: Tetrahydrofuran,

HATU: 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethylurea hexafluorophosphate,

DIEA: N,N-Diisopropylethylamine

M: molarity.

Using Fourier transform type NMR, ¹ H-NMR (proton nuclear magnetic resonance spectrum) was measured. For analysis, use ACD/SpecManager etc. Peaks for very light protons (eg, hydroxyl, amino, etc.) are not described.

MS (Mass Spectrometry) was measured using 1C/MS (Liquid Chromatography Mass Spectrometer). As the ionization method, an ESI (Electrospray Ionization) method or the like is used. The data represent those observed values. Typically, molecular ion peaks are observed. In the case of salts, molecular ion peaks or fragment ion peaks are usually observed in free form.

The following are the structural formulas and preparation methods of some Z substituents:

(1) N-methyl-4-aminoimidazole

4-Nitroimidazole (1.41 g), potassium carbonate (2.5 g), methyl iodide (1.9 g) and 20 mL of acetonitrile were reacted under reflux for 12 h. Spin to dryness under reduced pressure, add 50 mL of water, extract three times with 50 mL of ethyl acetate respectively, combine the organic layers, wash with saturated brine, add anhydrous Na ₂ SO ₄ to the organic phase and dry, and dry under reduced pressure to obtain 1-methyl-4 - Nitroimidazole (1.5 g).

1-Methyl-4-nitroimidazole (1 g), Pd/C (0.1 g) and 20 ml of ethanol were replaced with air in a hydrogen balloon, and the reaction was carried out at room temperature for about 24 h. The celite was filtered under reduced pressure, and the filtrate was dried under reduced pressure to obtain the title product (0.75 g).

MS(ESI ⁺ ): [M+H]+98.0.

(2) 1-(2-Methoxyethyl)-1H-pyrazol-4-amine

Using the same method as Reference Example 1, the title compound was obtained from 4-nitropyrazole, 2-bromomethylethyl ether, Pd/C.

MS(ESI ⁺ ): [M+H] ⁺ 142.0.

(3) 2-(4-Amino-1H-pyrazol-1-yl)ethanol

In the same manner as in Reference Example 1, the title compound was obtained from 4-nitropyrazole, 2-bromoethanol, Pd/C.

MS(ESI ⁺ ): [M+H] ⁺ 128.0.

(4) N,N-Dimethyl-2-(4-amino-1H-pyrazol-1-yl)ethylamine

Using the same method as Reference Example 1, the title compound was obtained from 4-nitropyrazole, N,N-dimethyl-2-bromo-ethylamine, Pd/C.

MS(ESI ⁺ ): [M+H] ⁺ 154.1.

(5) N-methyl-2-(4-amino-1H-pyrazol-1-yl)acetamide

Using the same method as Reference Example 1, the title compound was obtained from 4-nitropyrazole, N-methyl-2-bromo-acetamide, Pd/C.

MS(ESI ⁺ ): [M+H] ⁺ 154.0.

(6) 4-(4-Morpholinyl)aniline

4-Fluoronitrobenzene (1g), K ₂ CO ₃ (1.1g) and 5ml DMSO were stirred at room temperature for 30min, morpholine (0.6g) was added dropwise, heated at 120°C and stirred for 2h. The mixture was poured into alcohol and mixed with water In 20 mL of liquid (1:1), the yellow precipitate was filtered to obtain 1.4 g of 4-(4-nitrophenyl)morpholine. 4-(4-Nitrophenyl)morpholine (1 g), Pd/C (0.1 g) and 20 ml of ethanol were replaced with air in a hydrogen balloon, and the reaction was carried out at room temperature for about 24 h. The celite was filtered under reduced pressure, and the filtrate was dried under reduced pressure to obtain the title product (0.85 g). MS(ESI ⁺ ): [M+H] ⁺ 179.1.

The following are the preparation methods of some compounds of the application:

Example 1

N-(3-(6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)acrylamide

A) 4-Chloro-6-(3-nitrophenyl)pyrimidine

At room temperature, to 3-bromonitrobenzene (1 g) and 50 mL of anhydrous 1,4 dioxane were added bispinacol boronate (1.34 g) and KOAc (1.48 g) over catalyst Pd ( PPh ₃ ) ₄ (3%) was reacted with anaerobic conditions at 100°C for 12h. Cool to room temperature, add 4,6-dichloropyrimidine (1.07g), 2M K ₂ CO ₃ aqueous solution and Pd(PPh ₃ ) ₄ (3%) to the mixture and heat to 100°C to ensure anaerobic conditions to continue the reaction for 12h. Cooled to room temperature, filtered through celite, dried under reduced pressure, and purified by silica gel column chromatography to obtain the title compound (0.93 g).

¹ H NMR (300 MHz, CDCl ₃ ) δ 9.11 (s, 1H), 8.95 (s, 1H), 8.45 (d, J=8.0 Hz, 1H), 8.41 (d, J=9.3 Hz, 1H), 7.85 (s, 1H), 7.74(t, J=8.0Hz, 1H).

B) 6-(3-Nitrobenzene)-N ₄ -(1-(2-methoxyethyl)-1H-pyrazol-4-yl)-4-aminopyrimidine

To 4-chloro-6-(3-nitrophenyl)pyrimidine (0.3 g) and 10 mL of isopropanol at room temperature, was added 1-(2-methoxyethyl)-1H-pyrazole-4- Amine (0.24g) and DIEA (0.41g) were reacted under reflux for 6h, cooled to room temperature, precipitated out, filtered with suction and washed to obtain the title compound.

¹ H NMR (300MHz, CDCl3) δ 8.74 (s, 1H), 8.70 (s, 1H), 8.34-8.22 (m, 2H), 7.78 (s, 1H), 7.61 (t, J=8.0Hz, 1H ), 7.56(s, 1H), 7.50(s, 1H), 6.93(s, 1H), 4.31(t, J=5.1Hz, 2H), 3.77(t, J=5.1Hz, 2H), 3.35(s) , 3H).

C) 6-(3-Aminobenzene)-N ₄ -(1-(2-methoxyethyl)-1H-pyrazol-4-yl)-4-aminopyrimidine

Add 4-chloro-6-(3-nitrobenzene)-N ₄ -(1-(2-methoxyethyl)-1H-pyrazol-4-yl)-4-aminopyrimidine ( 0.2g), 0.02g Pd/C, and 20mL EtOH, under the protection of N ₂ , heated to reflux, slowly added dropwise 85% hydrazine hydrate (0.32g) within 30min, the reaction was completed dropwise, suction filtered, and the filtrate was dried under reduced pressure The title compound was obtained.

MS(ESI ⁺ ): [M+H] ⁺ 311.2

D) N-(3-(6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)acrylamide

6-(3-Aminobenzene)-N ₄ -(1-(2-methoxyethyl)-1H-pyrazol-4-yl)-4-aminopyrimidine (0.04g), 10mL was added to the reaction flask Dry DCM, add 1 mL of acryloyl chloride diluted in dry DCM dropwise at 0°C, stir for 30 min, and filter with suction to obtain the title compound.

¹ H NMR (300MHz, DMSO) δ 8.69 (s, 1H), 8.48 (s, 1H), 8.06 (s, 1H), 7.81 (d, J=7.7Hz, 1H), 7.73 (d, J=7.5 Hz, 1H), 7.59(s, 1H), 7.47(t, J=7.8Hz, 1H), 7.10(s, 1H), 6.49(dd, J=16.9, 10.0Hz, 1H), 6.31(d, J =16.7Hz, 1H), 5.78(d, J=10.6Hz, 1H), 4.25(t, J=4.9Hz, 2H), 3.68(t, J=4.9Hz, 2H), 3.23(s, 3H)

MS(ESI)m/z 365.2[M+H]+.

Examples 2 to 23

In Examples 2 to 23, the same method as in Example 1 was used.

Wherein Examples 3, 5, 7, 9, 15, 16, 19 and 20 in step A) correspondingly replaced 3-bromonitrobenzene (1 g) with 4-bromonitrobenzene (1 g) and other conditions remained unchanged.

According to the structural formula of the example, its step B) corresponds to different aminoalkylated substituents, which are specifically the Z substituents (described above).

Wherein Examples 13-23 correspond to the dropwise addition of 1 mL of acryloyl chloride diluted with dry DCM in step D), and the dropwise addition of 1 mL of dry DCM diluted crotonyl chloride at 0 °C, and other conditions remain unchanged.

The above molar equivalent ratio is the same as the corresponding reaction equivalent in Example 1.

The following title compounds can then be obtained, see Table 2. MS in this table refers to the measured value.

Table 2 Part of the compound structure and NMR data

Example 24

2-cyano-N-(3-(6-((1-methyl-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)acetamide

Experimental steps A), B), C) Using the same method as in Example 1, 6-(3-aminobenzene)-N ₄ -(1-methyl-1H-pyrazol-4-yl)-4 can be obtained -Aminopyrimidine.

E) 2-cyano-N-(3-(6-((1-methyl-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)acetamide

6-(3-Aminobenzene)-N ₄ -(1-methyl-1H-pyrazol-4-yl)-4-aminopyrimidine (0.12 g, ), cyanoacetic acid (0.04 g), DIEA (0.13 g ) and 15 mL of dry THF, HATU (0.37 g) was added under stirring at 0° C. to stir the reaction for 30 min, and then the reaction was stirred at room temperature. After the reaction was completed, 20 mL of ethyl acetate was added, and 10% aqueous citric acid solution was added, and the mixture was washed twice with a saturated NaHCO ₃ aqueous solution. The organic phase was mixed and purified by silica gel column chromatography to obtain the title compound.

¹ H NMR (300MHz, DMSO) δ 8.66 (s, 1H), 8.33 (s, 1H), 8.02 (s, 1H), 7.73 (d, J=7.5Hz, 1H), 7.64 (d, J=7.1 Hz, 1H), 7.53(s, 1H), 7.47(t, J=7.9Hz, 1H), 7.07(s, 1H), 3.94(s, 2H), 3.83(s, 2H)

MS(ESI ⁺ ): [M+H] ⁺ 334.2.

Examples 25 to 32

In Examples 25 to 32, the same method as in Example 24 was used.

Wherein in Examples 26, 27 and 30-32, in step A), 3-bromonitrobenzene (1 g) is correspondingly replaced with 4-bromonitrobenzene (1 g) and other conditions remain unchanged.

According to the structural formula of the example, its step B) corresponds to different aminoalkylated substituents, which are specifically the Z substituents (described above).

The above molar equivalent ratio is the same as the corresponding reaction equivalent in Example 1.

The following title compounds can then be obtained, see Table 2. MS in this table refers to the measured value.

Table 2 Part of the compound structure and NMR data

Example 33

N-[2-oxo-2-((3-(6-(1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)-phenyl -amino)ethyl]acrylamide

Experimental steps A), B), C) Using the same method as in Example 1, 6-(3-aminobenzene)-N ₄ -(1-(2-methoxyethyl)-1H-pyrazole can be obtained -4-yl)-4-aminopyrimidine.

F) 6-[3-(Carbamate, N-(2-oxo-2-(phenylamino)ethyl)-, 1,1-dimethylethyl ester)]-N ₄ -(1- Methyl-1H-pyrazol-4-yl)-4-aminopyrimidine

6-(3-Aminobenzene)-N4-(1-methyl-1H-pyrazol- ₄ -yl)-4-aminopyrimidine (0.2 g) was dissolved in 10 mL DCM at room temperature, EDCI (0.36 g) was added ), HOBt (0.25g), DIEA (0.34g), the reaction was stirred at 0°C for 30min, 6-(3-aminobenzene)-N ₄ -(1-(2-methoxyethyl)-1H-pyrazole was added -4-yl)-4-aminopyrimidine, then transferred to room temperature and stirred for 6 h. After the reaction was completed, 20 mL of DCM was added, and NaHCO ₃ aqueous solution was added to wash twice, the organic phase was mixed, and separated and purified by silica gel column chromatography to obtain the title compound.

¹ H NMR (300 MHz, CDCl ₃ ) δ 9.05 (s, 1H), 8.53 (s, 2H), 7.91 (s, 1H), 7.78 (s, 1H), 7.71-7.58 (m, 1H), 7.55 ( s, 1H), 7.50-7.40 (m, 1H), 7.30-7.15 (m, 1H), 6.74 (s, 1H), 6.04 (s, 1H), 4.24 (t, J=5.0Hz, 2H), 3.95 (s, 2H), 3.71 (t, J=4.9Hz, 2H), 3.29 (s, 3H), 1.40 (s, 9H).

G) 6-[3-(2-(Amino)acetanilide)]-N ₄ -(1-(2-methoxyethyl)-1H-pyrazol-4-yl)-4-pyrimidinamine

6-[3-(Carbamate, N-(2-oxo-2-(phenylamino)ethyl)-,1,1-dimethylethyl ester)]-N ₄ -(1-methyl) -1H-pyrazol-4-yl)-4-aminopyrimidine (0.5 g) was dissolved in 2 mL of DCM, cooled to 0° C. and slowly added TFA (2 mL), and the reaction was stirred at room temperature for 5 h. After the reaction, it was dried under reduced pressure, and the title compound was obtained by slurring with ether and suction filtration.

MS(ESI ⁺ ): [M+H] ⁺ 368.2.

H) Use the same method as D) or E) in Example 1 to obtain the title compound. Molar equivalents and other reaction conditions were unchanged.

¹ H NMR (300MHz, DMSO) δ 8.83(s, 1H), 8.57(s, 1H), 8.28(s, 1H), 8.11(s, 1H), 7.74(d, J=15.0Hz, 2H), 7.63-7.53 (m, 2H), 7.49 (s, 1H), 6.33 (dd, J=15.2, 8.2Hz, 1H), 6.12 (d, J=16.8Hz, 1H), 5.63 (d, J=8.9Hz) , 1H), 4.28(s, 2H), 4.04(s, 2H)., 3.68(s, 2H), 3.23(s, 3H).

MS(ESI ⁺ ): [M+H] ⁺ 422.2.

Examples 34 to 39

In Examples 34 to 39, using the same method as Example 33, the title compounds were obtained, see Table 3. MS in this table refers to the measured value.

Table 3 Partial compound structures and NMR data

Example 40: Experimental Example (JAK-1 and JAK-3 Enzyme Inhibition Assay)

Purpose:

Compounds were evaluated for inhibition of JAK-1 and JAK-3 kinases.

Experimental principle:

Different proteolysis of specific phosphorylated and non-phosphorylated polypeptide substrates based on fluorescence resonance energy transfer (FRET) coupled proteolytic enzymes. The two ends of the polypeptide substrate are respectively labeled as FRET energy donor coumarin and energy acceptor fluorescein. At this time, when the distance between the donor and the acceptor is relatively close, the donor can be excited and the energy transfer can occur.

In the kinase reaction (Kinase Reaction), JAK-1 or JAK-3 can transfer the γ-phosphate in ATP to a single tyrosine residue of the polypeptide substrate, if there is a JAK-1 or JAK-3 inhibitor in the system , the γ-phosphate group on ATP will not be transferred to the substrate polypeptide, and the phosphorylation reaction cannot occur. Based on this principle, an evaluation experiment for kinase inhibitors was designed. The substrate polypeptide is designed with a kinase phosphorylation site, which is also a protease cleavage site. Two fluorescent groups are connected at both ends, which are the donor and the acceptor. The reaction system If the kinase activity is maintained, the γ-phosphate group will be transferred to the enzyme cleavage site of the substrate, so that it will not be cleaved by protease and separated into two sections. Under the excitation of a specific wavelength of laser, the fluorescence energy of one section will be transferred to the other. A fluorophore at one end emits energy. Otherwise, after the enzymatic activity is inhibited, the phosphate group cannot be transferred, the enzyme cleavage site of the substrate will be cleaved by the enzyme in the system, and the substrate will be separated into two sections, and no fluorescence energy transfer will occur. Based on this, the activity of the kinase was evaluated.

Experimental steps:

In this experiment, a 10 μl kinase reaction system was selected. In the JAK1 inhibitor screening, the system contains JAK1 kinase concentration of 1ng/μl, ATP 3.92 μM; JAK2 inhibitor screening, JAK2 kinase 0.004ng/μl, ATP 3.96 μM; JAK3 inhibitor screening , each system contains JAK3 kinase 0.012ng/μl, ATP 1.43μM. The TK Substrate-biotin substrate is 1 μM, and the primary screening concentration of the compound is 10 μM.

(1) Preparation of kinase reaction buffer

Add 1ml of 5×Kinase Buffer to 4ml of double-distilled water to dilute to 1×, add 5μl of 1M DTT and 25μl of 1M MgCl2, named Kinase Buffer and store at room temperature.

(2) Configuration of compounds to be screened

The sample compounds were dissolved in DMSO according to the mass to prepare a stock solution with a concentration of 100 mM, and the compounds were diluted and prepared with kinase reaction buffer, and the final reaction concentration of the compounds was 10 μM.

(3) Reaction system

The experimental carrier plate is a black 384-well microplate, and the reaction system is 10 μl.

Fluorescence detection

Under excitation at 330 nm, emission intensities at 665 nm and 620 nm were detected.

The microplate reader automatically calculates Ratio=665/620*10000

(5) Data analysis

Calculate the inhibition rate for each sample as follows

Inhibition rate=(Ratio MAX-Ratio sample)/(Ratio MAX-Ratio NEG)*100

Table 4 Experimental results

The results showed that several compounds exhibited certain inhibitory effects on JAK kinase.

IC ₅₀ Evaluation

The IC ₅₀ value of the compound that showed better activity was evaluated after its concentration was diluted, and it was found that the compound of Example 5 showed a highly active JAK-3 inhibitory effect (IC ₅₀ =1.7nm), a moderately strong selective activity (JAK -1/JAK-3=586, JAK-2/JAK-3=60).

Claims (10)

1. A substituted phenylpyrimidine derivative having general formula (I) as a JAK kinase inhibitor or a pharmaceutically acceptable salt thereof: in, X is a nitrogen atom or -CH-; Y is a nitrogen atom or -CH-; Z is Substituted or unsubstituted C _3-8 heterocyclic alkyl; W is a substituted 5-membered heteroaryl; ^R1 is in, V is a hydrogen atom, halogen or cyano; R ⁴ , R ⁵ , R ⁶ and R ⁷ are respectively a hydrogen atom, a substituted or unsubstituted C _1-6 hydrocarbyl group, and a substituted or unsubstituted C _1-6 heterohydrocarbyl group; R ² is a hydrogen atom, a halogen atom, a substituted or unsubstituted 4, 5, 6, 7- or 8-membered heterocycle; R ³ is substituted C _1-6 hydrocarbyl, substituted or unsubstituted C _1-6 heterohydrocarbyl. 2. The substituted phenylpyrimidine derivative having the general formula (I) according to claim 1 as a JAK kinase inhibitor or a pharmaceutically acceptable salt thereof, characterized in that, Z is 3. The substituted phenylpyrimidine derivative having the general formula (I) according to claim 2 as a JAK kinase inhibitor or a pharmaceutically acceptable salt thereof, wherein the R ² is a hydrogen atom, a fluorine atom , substituted or unsubstituted 4, 5 or 6-membered nitrogen-containing heterocycles. 4. The substituted phenylpyrimidine derivative with general formula (I) according to claim 2 as a JAK kinase inhibitor or a pharmaceutically acceptable salt thereof, wherein the W is a nitrogen-containing five-membered heteroaromatic ring. 5. The substituted phenylpyrimidine derivative having the general formula (I) as claimed in claim 2 as a JAK kinase inhibitor or a pharmaceutically acceptable salt thereof, wherein the ^R is methyl or substituted The C _1-6 alkyl group of the C _1-6 alkyl group is a halogen atom, a hydroxyl group, a C _1-6 alkoxy group, an aliphatic amino group, and an amide group. 6. The substituted phenylpyrimidine derivative with general formula (I) according to claim 1 is used as a JAK kinase inhibitor or a pharmaceutically acceptable salt thereof, which is any of the following: N-(3-(6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)acrylamide, N-(3-(6-((1-Methyl-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)acrylamide, N-(4-(6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)acrylamide, N-(3-(2-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)acrylamide, N-(4-(2-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)acrylamide, N-(3-(6-(4-morpholinylamino)pyrimidin-4-yl)phenyl)acrylamide, N-(4-(6-(4-morpholinylamino)pyrimidin-4-yl)phenyl)acrylamide, N-(3-(2-(4-morpholinylamino)pyrimidin-4-yl)phenyl)acrylamide, N-(4-(2-(4-morpholinylamino)pyrimidin-4-yl)phenyl)acrylamide, N-(3-(6-((1-(2-N,N-dimethylethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)acrylamide, N-(3-(6-((1-(N-methylacetamide)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)acrylamide, N-(3-(6-((1-(2-hydroxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)acrylamide, N-(3-(6-((1-Methyl-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)-2-butenamide, N-(3-(6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)-2-butenamide, N-(4-(6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)-2-butenamide, N-(4-(2-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)-2-butenamide, N-(3-(6-(4-morpholinylamino)pyrimidin-4-yl)phenyl)-2-butenamide, N-(3-(2-(4-morpholinylamino)pyrimidin-4-yl)phenyl)-2-butenamide, N-(4-(6-(4-morpholinylamino)pyrimidin-4-yl)phenyl)-2-butenamide, N-(4-(2-(4-morpholinylamino)pyrimidin-4-yl)phenyl)-2-butenamide, N-(3-(6-((1-(2-N,N-dimethylethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)-2-butane enamide, N-(3-(6-((1-(N-methylacetamide)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)-2-butenamide, N-(3-(6-((1-1-(2-hydroxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)-2-butenamide, N-(3-(6-((1-1-(2-hydroxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)-2-butenamide, 2-cyano-N-(3-(6-((1-methyl-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)acetamide, 2-cyano-N-(3-(6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)acetamide, 2-cyano-N-(4-(6-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)acetamide, 2-cyano-N-(4-(2-((1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)acetamide, 2-cyano-N-(3-(((6-(4-morpholinoanilino)pyrimidin-4-yl)amino)methyl)phenyl)acetamide, 2-cyano-N-(3-(((2-(4-morpholinylamino)pyrimidin-4-yl)amino)methyl)phenyl)acetamide, 2-cyano-N-(4-(((6-(4-morpholinoanilino)pyrimidin-4-yl)amino)methyl)phenyl)acetamide, 2-cyano-N-(4-(((2-(4-morpholinoanilino)pyrimidin-4-yl)amino)methyl)phenyl)acetamide, N-[2-oxo-2-((3-(6-(1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)-phenyl -amino)ethyl]acrylamide, N-[2-oxo-2-((3-(6-(1-(2-methoxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)-phenyl -amino)ethyl]butenamide, N-[2-oxo-2-((3-(6-(4-morpholinoanilino)amino)pyrimidin-4-yl)-phenyl-amino)ethyl]acrylamide, N-[2-oxo-2-((3-(6-(4-morpholinoanilino)amino)pyrimidin-4-yl)-phenyl-amino)ethyl]butenamide, 2-cyano -N-[2-oxo-2-((3-(6-(1-methyl-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)-phenyl-amino)ethyl] acetamide, 2-cyano-N-[2-oxo-2-((3-(6-(4-morpholinylamino)amino)pyrimidin-4-yl)-phenyl-amino)ethyl]acetamide, 2-cyano-N-[2-oxo-2-((3-(6-(1-(N-methylacetamide)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl )-phenyl-amino)ethyl]acetamide, 2-cyano-N-(4-(6-((1-1-(2-hydroxyethyl)-1H-pyrazol-4-yl)amino)pyrimidin-4-yl)phenyl)acetamide. 7. A pharmaceutical composition containing a therapeutically effective amount of one or more substituted phenylpyrimidine derivatives of general formula (I) as described in any one of claims 1-6 as JAK kinases An inhibitor or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or adjuvant. 8. The preparation method of the substituted phenylpyrimidine derivative with general formula (I) as claimed in claim 1 as a JAK kinase inhibitor or a pharmaceutically acceptable salt thereof, comprising the steps of: in, Step 1: Compound III is converted into compound IV under the conditions of tetrakistriphenylphosphine palladium and basic potassium acetate with double pinacol boronate; Step 2: add 2,4-chloro-pyrimidine or 4,6-chloro-pyrimidine, tetrakistriphenylphosphine palladium and potassium carbonate aqueous solution to the reaction solution of step 1 to carry out suziki reaction to obtain compound V; Step 3: Compound V is reacted with a substituted amine to obtain Compound VI; Step 4: the reduction reaction of compound VI, into compound VII; Step 5: Compound VII is subjected to an amide-forming reaction with a R ¹ substitution reagent to obtain the compound of the present invention. 9. The substituted phenylpyrimidine derivative with general formula (I) of any one of claims 1-6 is used as a JAK kinase inhibitor or a pharmaceutically acceptable salt thereof in the preparation and treatment of autoimmune diseases, Sjogren Medicinal use in medicament for Behcet's disease, multiple sclerosis, systemic lupus erythematosus. 10. The use according to claim 9, wherein the autoimmune disease is psoriasis, rheumatoid arthritis or inflammatory bowel disease.