Compound with terminal alkynyl structure and application thereof

Abstract

The invention relates to a compound with a terminal alkynyl structure as shown in formula (I), a stereoisomer, a tautomer or pharmaceutically acceptable salt thereof, methods related to preparation and application of the compound, a pharmaceutical composition containing the compound and a related method for regulating immune system diseases. The compound shows an excellent effect of regulating immune system diseases, and has a wide application prospect in the field of regulating immune system disease treatment.

Description

A compound with terminal alkynyl structure and its application Technical Field

[0001] The present invention relates to a compound with a terminal alkynyl structure, a preparation method thereof, and application of the compound in disease treating drugs. Background Art

[0002] Autoimmune diseases, including rheumatoid arthritis (RA), psoriasis, and alopecia areata (AA), are a broad category of diseases caused by tissue damage resulting from an immune response to self-antigens. These diseases are characterized by unclear pathogenesis, limited targets, and a lack of effective therapeutic agents. Currently, several small-molecule JAK inhibitors are marketed for the treatment of various autoimmune diseases. However, with their clinical application, side effects such as opportunistic infections and anemia have gradually emerged. These side effects are primarily due to the non-selective inhibition of different JAK kinase subtypes. This non-selective inhibition of JAKs inevitably affects the physiological effects of various cytokines, leading to corresponding side effects.

[0003] Early JAK inhibitors, whether non-selective or JAK1-selective, have JAK1 inhibitory activity, which is associated with serious side effects such as severe infection, mortality, malignancy, adverse cardiovascular events, and thrombosis risk. Therefore, the FDA has issued black box warnings for JAK inhibitors used to treat chronic inflammation (such as tofacitinib, baricitinib, upadacitinib, and abrocitinib). The structural formulas of these inhibitors are as follows:

[0004] Subsequently, based on tofacitinib and other compounds, optimized compounds, represented by ritlecitinib, have shown better selective inhibition of JAK3, thereby reducing toxic side effects, but the inhibitory activity is general. The structural formula of ritlecitinib is as follows:

[0005] Inspired by the lack of an obvious structure-activity relationship between the excellent efficacy (inhibitory effect), selectivity, pharmacokinetic properties, and safety of the drug compound and its structure, the inventors surprisingly discovered that by substituting a terminal alkynyl group at position 7 on the 7-deazapurine ring, the resulting compound exhibited JAK3 inhibition far superior to that of ritlecitinib, with high inhibitory selectivity, far superior to that of tofacitinib. Therefore, the compounds of the present invention are expected to achieve excellent clinical treatment for autoimmune diseases such as alopecia areata and rheumatoid arthritis mediated by B and T cells, allowing patients with autoimmune diseases to enjoy better therapeutic effects while having fewer toxic side effects.

[0006] Summary of the Invention

[0007] The present invention provides a compound whose structure cannot be synthesized from the structures of ritlecitinib and tofacitinib, which has a much better JAK3 inhibitory effect than ritlecitinib and a much better selectivity for inhibiting JAK kinase subtypes than tofacitinib.

[0008] Specifically, the present invention provides a compound of formula (I), or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof,

[0009] Wherein, X is an oxygen atom, -NH-;

[0010] Y is a nitrogen atom, -C(H)=;

[0011] Z is a nitrogen atom, CR 1 ;

[0012] Q is a bond, an oxygen atom, -O-CH2-, -NH-, -CH2-, -NH-CH2-, C1-C6 alkyl, C2-C8 alkynyl, C2-C8 alkynylC1-C6 alkyl, C1-C6 alkylC2-C8 alkynyl, C2-C8 alkenyl, C2-C8 alkenylC1-C6 alkyl, C1-C6 alkylC2-C8 alkenyl, aryl, C2-C8 alkynylaryl, arylC2-C8 alkynyl, or heteroaryl;

[0013] R 0 , R 1 , R 3 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 Each is independently hydrogen, deuterium, halogen, alkyl, deuterated alkyl, haloalkane, cycloalkyl, heterocyclyl, heterocyclylalkyl, aryl, heteroaryl, hydroxy, amino, sulfone, phosphorus carbonyl, substituted silicon, alkenyl, alkynyl, wherein said alkyl, aryl, heteroaryl are independently optionally substituted with one or more substituents selected from the group consisting of C1-C6 alkyl, halogen, hydroxy, methoxy, amino, cyano, alkylamino, dialkylamino, trifluoromethyl, C3-C6 cycloalkyl, C2-C8 alkynyl, or C2-C8 alkenyl;

[0014] n1 = 0, 1, 2, 3, or 4;

[0015] n2=0, 1, 2, 3, or 4.

[0016] The present invention provides a compound of formula (II), or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof,

[0017] Wherein, Y is a nitrogen atom, -C(H)=;

[0018] Z is a nitrogen atom, CR 1 ;

[0019] Q is a bond, an oxygen atom, -O-CH2-, -NH-, -CH2-, -NH-CH2-, C1-C6 alkyl, C2-C8 alkynyl, C2-C8 alkynylC1-C6 alkyl, C1-C6 alkylC2-C8 alkynyl, C2-C8 alkenyl, C2-C8 alkenylC1-C6 alkyl, C1-C6 alkylC2-C8 alkenyl, aryl, C2-C8 alkynylaryl, arylC2-C8 alkynyl, or heteroaryl;

[0020] R 1 , R 3 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 Each is independently hydrogen, deuterium, halogen, alkyl, deuterated alkyl, haloalkane, cycloalkyl, heterocyclyl, heterocyclylalkyl, aryl, heteroaryl, alkenyl, alkynyl, wherein the alkyl, aryl, heteroaryl are independently optionally substituted by one or more substituents selected from the following: C1-C6 alkyl, halogen, hydroxy, methoxy, amino, cyano, alkylamino, dialkylamino, trifluoromethyl, C3-C6 cycloalkyl, C2-C8 alkynyl, or C2-C8 alkenyl.

[0021] The present invention provides a compound of formula (III), or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof,

[0022] Where Z is a nitrogen atom, or CR 1 ;

[0023] R 1 , R 3 , R 8 , R 9, R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 Each is independently hydrogen, deuterium, halogen, alkyl, deuterated alkyl, haloalkane, cycloalkyl, heterocyclyl, heterocyclylalkyl, aryl, heteroaryl, alkenyl, alkynyl, alkenylalkyl, alkynylalkyl, wherein the alkyl, aryl, heteroaryl are independently optionally substituted with one or more substituents selected from the group consisting of C1-C6 alkyl, halogen, hydroxy, methoxy, amino, cyano, alkylamino, dialkylamino, trifluoromethyl, C3-C6 cycloalkyl, C2-C8 alkynyl, or C2-C8 alkenyl.

[0024] Preferably, the present invention provides the following compounds, or stereoisomers, tautomers or pharmaceutically acceptable salts thereof,

[0025] For example, the present invention provides the following compounds, or stereoisomers, tautomers or pharmaceutically acceptable salts thereof,

[0026] The present invention provides a pharmaceutical composition comprising a therapeutically effective dose of any one of the compounds of the present invention or its stereoisomers, tautomers or pharmaceutically acceptable salts, and a pharmaceutically acceptable carrier.

[0027] The present invention provides the use of a compound described herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition described herein, in the preparation of a medicament for treating a Janus kinase (JAK)-related disease. Furthermore, the present invention provides a method for treating a disease by inhibiting Janus kinase (JAK), comprising administering to a subject in need thereof a therapeutically effective dose of a compound described in any one of claims 1 to 4, or a stereoisomer, tautomer, or pharmaceutically acceptable salt thereof.

[0028] Preferably, the disease of the present invention is rheumatoid arthritis, alopecia areata, vitiligo, asthma, chronic obstructive pulmonary disease, tumor, Crohn's disease, inflammatory bowel disease such as ulcerative colitis, proctitis, eosinophilic gastroenteritis or mastocytosis.

[0029] Detailed Description of the Invention

[0030] All technical and scientific terms used in this specification have the same meanings as commonly understood by those skilled in the art.

[0031] The term "alkyl" refers to a saturated aliphatic hydrocarbon group having 1 to 10 carbon atoms in this article, and the term includes straight and branched chain hydrocarbon groups. Non-limiting examples of alkyl include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, neopentyl, n-hexyl, etc. Alkyl described herein can be optionally substituted with one or more of the following substituents: deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, hydroxyl, carboxyl, amino, alkyl, alkoxy, acyl, acyloxy, oxo, amide, ester, amido, cycloalkyl, cycloalkenyl, heterocycloalkyl, alkenyl, alkenyloxy, alkynyl, cycloalkyloxy, heterocycloalkyloxy, aryloxy, heteroaryloxy, aryl or heteroaryl.

[0032] The term "aryl" herein refers to a 6-10 membered all-carbon monocyclic or fused polycyclic (i.e., rings that share adjacent pairs of carbon atoms) group, a polycyclic (i.e., rings with adjacent pairs of carbon atoms) group having a conjugated π electron system. The aryl group can be covalently attached to the defined chemical structure at any carbon atom that produces a stable structure. The aryl groups described herein may be optionally substituted with one or more of the following substituents: fluorine, chlorine, bromine, iodine, cyano, nitro, hydroxyl, carboxyl, amino, alkyl, alkoxy, acyl, amide, ester, amine, sulfonyl, sulfinyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, alkenyl, alkynyl, and cycloalkoxy.

[0033] The term "heterocyclic group" refers to a ring system containing a nitrogen atom or an oxygen atom. The ring system can be "parallel" to aromatic and non-aromatic ring systems, or linked to other ring systems through "spiro carbon atoms", such as the following structure:

[0034]

[0035] etc.

[0036] The term "heteroaryl" as used herein refers to an aromatic group consisting of 5 to 10 atoms and containing at least one heteroatom selected from N, O, or S. The term can have a single ring (non-limiting examples include furan, thiophene, imidazole, pyrazole, pyridine, pyrazine, oxazole, thiazole, etc.) or multiple fused rings (non-limiting examples include benzothiophene, benzofuran, indole, isoindole, etc.), wherein the fused rings may or may not be aromatic groups containing heteroatoms, provided that the point of attachment is through an atom of the aromatic heteroaryl group. The heteroaryl groups described herein may be optionally substituted with one or more of the following substituents: fluoro, chloro, bromo, iodo, cyano, nitro, hydroxy, amino, alkyl, alkoxy, acyl, acyloxy, amide, ester, amine, sulfonyl, sulfinyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, alkenyl, alkynyl, and cycloalkoxy.

[0037] The term "alkenyl" refers to an alkenyl group having 2 to 8 carbon atoms and at least one alkenyl unsaturated site in this article. Non-limiting examples of alkenyl include vinyl, propenyl, allyl, isopropenyl, butenyl, isobutenyl etc. Alkenyl described herein can be optionally substituted with one or more of the following substituents: deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, hydroxyl, carboxyl, amino, alkyl, alkoxy, acyl, amide, ester group, amino, sulfonyl, sulfinyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, cycloalkyloxy, sulfydryl, alkyl mercapto, deuterated alkyl mercapto, sulfone, sulfoxide, amino, silicon, phosphono, deuterated alkyl, heterocycloalkyl, aryl, heteroaryl, alkynyl, alkenyl, arylalkyl, ester group.

[0038] The term "alkynyl" is herein intended to refer to an alkyl radical in which two adjacent carbon atoms are connected by a triple bond, wherein the alkyl radical is as defined herein. Alkynyl refers to an unsaturated alkyl radical as defined above consisting of at least two carbon atoms and at least one carbon-carbon triple bond, such as ethynyl, 1-propynyl, 2-propynyl, 1-, 2- or 3-butynyl, etc. Alkynyl may be substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from deuterium, fluorine, chlorine, bromine, iodine, cyano, nitro, hydroxyl, carboxyl, amino, alkyl, alkoxy, acyl, amide, ester, amino, sulfonyl, sulfinyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, cycloalkyloxy, sulfhydryl, alkylthiol, deuterated alkylthiol, sulfone, sulfoxide, amino, silicon, phosphono, deuterated alkyl, heterocycloalkyl, aryl, heteroaryl, alkynyl, alkenyl, arylalkyl, ester.

[0039] The beneficial effects of the present invention include:

[0040] (1) The compounds of the present invention have a good inhibitory effect on JAK3. For example, the inhibitory activity IC of compounds 1, 2, 3, 4 and 5 on JAK3 is 50 =1.1~2.4nM, and the inhibitory activity of these compounds against JAK3 was significantly better than that of Ritlecitinib (PF-06651600) against JAK3 (IC 50 =33 nM).

[0041] (2) The compounds of the present invention have a selective inhibitory effect on JAK3, a kinase of the JAK family, and have no inhibitory effect on other kinase subtypes of the JAK family (JAK1, JAK2 and TYK2). For example, the inhibitory activity IC values ​​of compounds 2 and 3 of the present invention on other kinase subtypes of the JAK family (JAK1, JAK2 and TYK2) are 50>1000nM, while the marketed drug Tofacitinib does not have the above selectivity characteristics. This indicates that the compound of the present invention has potential safety characteristics superior to the marketed drug Tofacitinib in future clinical use.

[0042] (3) The inhibitory effect of the compounds of the present invention on the expression of phosphorylated STAT5 in CD3+ cells in human PBMCs, for example, the inhibitory activity IC of compounds 2 and 3 on the expression of phosphorylated STAT5 in CD3+ cells in human PBMCs is 50 =33~65nM, superior to Ritlecitinib (IC 50 =597nM).

[0043] (4) The in vivo pharmacodynamics of Compound 2 on CIA model rats was positively correlated with the dose, and the 30 mg / kg dose of Compound 2 was superior to the efficacy of Ritlecitinib at the same dose in alleviating hind limb arthritis symptoms in rats. The compounds of the present invention have high selectivity for JAK3 and are expected to achieve better clinical treatment for autoimmune diseases such as alopecia areata and rheumatoid arthritis mediated by B and T cells, allowing patients with autoimmune diseases to enjoy better therapeutic effects. Specific implementation method:

[0044] The present invention is further illustrated by the following examples, but the present invention is not limited thereto. Throughout this application, various examples of the compounds and methods of the present invention are mentioned herein. The present invention is not limited to these examples. The following examples are merely provided to provide methods for practicing the present invention and are not intended to limit the scope of the present invention in any way.

[0045] The compounds provided herein can be prepared by standard synthetic methods known in the art, and this specification provides general methods for preparing the compounds of the present invention. Starting materials can generally be obtained commercially or prepared by methods well known to those skilled in the art.

[0046] The process is as follows:

[0047] Using SM-1 as the starting material, a coupling reaction is carried out with SM2 to obtain compound (IM1), which is then coupled with SM-3 to obtain compound (IM2), deprotected to obtain compound (IM-3), and finally coupled with SM-4 to obtain compound II.

[0048] The compounds of the present invention and corresponding preparation methods are further explained and listed below by examples and preparations. It should be understood that although typical or preferred reaction conditions are given in the specific examples, those skilled in the art may also use other reaction conditions. Optimum reaction conditions may vary with the specific reaction substrate or solvent used, but the conditions can be determined by conventional optimization by those skilled in the art.

[0049] Intermediate preparation

[0050] Intermediate 1:

[0051] SM-1 (6.02 g) was dissolved in 50 mL of DMF. The temperature was cooled to 10°C, and N-iodosuccinimide (NIS, 7.88 g) was added portionwise. After the addition was complete, the temperature was raised to room temperature and the reaction was allowed to react for 3 hours. The reaction solution was slowly poured into 80 mL of 10-20°C water. After the addition was complete, the mixture was stirred at 10-20°C for 1 hour, then filtered. The filter cake was dried at 50°C to yield 9.51 g of an off-white solid. m / z: 313.90 (M+1).

[0052] Compound preparation

[0053] Example 1:

[0054] first step:

[0055] Intermediate-1 (1.5 g), cuprous iodide (91 mg), dichlorobis(triphenylphosphine)palladium (Pd(PPh3)Cl2) (670 mg), triethylamine (1.21 g), trimethylsilylacetylene (936 mg), and 15 mL of tetrahydrofuran were added to a reaction flask in that order. Under nitrogen, the temperature was raised to 35°C and the reaction was allowed to proceed for 2 h. The reaction mixture was passed through a celite funnel, and the filter cake was washed with 50 mL of ethyl acetate. The filtrate and washings were combined, concentrated, and purified on a silica gel column to yield 980 mg of a yellow solid. m / z: 284.00 (M+1).

[0056] Step 2:

[0057] Intermediate 1-1 (980 mg), SM-2 (889 mg), potassium carbonate (1.44 g), 12 mL of water, and 2.5 mL of methyl isobutyl ketone were added to a reaction flask and heated to 90°C for 4 h. The reaction solution was extracted with 15 mL x 2 of ethyl acetate. The organic phase was washed with 20 mL of saturated sodium chloride, dried over sodium sulfate, concentrated, and purified on a silica gel column to yield 680 mg of a yellow solid. m / z: 496.20 (M+1).

[0058] Step 3:

[0059] Intermediate 1-2 (600 mg) and 12 mL of acetonitrile were added to the reaction flask. After stirring at room temperature until dissolved, trimethylsilyl iodide (533 mg) was added. After stirring at room temperature for 1 hour, triethylamine (355 mg) was added and the mixture was stirred for 15 minutes. The mixture was concentrated and purified on a silica gel column to obtain 1.0 g of a yellow solid. m / z: 362.15 (M+1).

[0060] Step 4:

[0061] Intermediate 1-3 (1.0 g), lithium hydroxide (256 mg), 10 mL of tetrahydrofuran, and 5 mL of water were added to the reaction flask in that order and stirred at room temperature for 1.5 h. The reaction solution was extracted with 15 mL x 2 of ethyl acetate. The organic phase was washed with 20 mL of saturated sodium chloride, dried over sodium sulfate, concentrated, and purified on a silica gel preparative plate to yield 200 mg of a yellow solid. m / z: 290.10 (M+1).

[0062] Step 5:

[0063] Intermediate 1-4 (200 mg) and 5 mL of N,N-dimethylformamide were added to a reaction flask. After stirring at room temperature, diisopropylethylamine (DIPEA, 268 mg) and acryloyl chloride (32 mg) were added sequentially. After stirring at room temperature for 0.5 h, the reaction mixture was added with 15 mL of water and extracted with 10 mL × 3 of ethyl acetate. The organic phase was washed with 15 mL of saturated sodium chloride, dried over sodium sulfate, concentrated, and purified on a silica gel preparative plate to yield 26 mg of a light yellow solid. m / z: 344.10 (M+1).

[0064] Example 2:

[0065] first step:

[0066] SM-1 (2.0 g), p-dimethylaminopyridine (DMAP) (87.4 mg), triethylamine (2.17 g), and 40 mL of dichloromethane were added to a reaction flask in that order, followed by p-toluenesulfonyl chloride (1.64 g). The mixture was allowed to react at room temperature for 2 h. The reaction solution was concentrated under reduced pressure, then added with 10 mL of ethanol. The mixture was stirred at room temperature for 0.5 h, then in an ice-water bath for 0.5 h. The mixture was then filtered, the filter cake rinsed with petroleum ether, and air-dried to yield 3.2 g of a yellow solid. m / z: 433.85 (M+1).

[0067] Step 2:

[0068] Intermediate 2-1 (3.2 g), SM-2 (2.65 g), triethylamine (2.17 g), and 31 mL of acetonitrile were added to a reaction flask and heated to 95°C for 4 h. The reaction solution was concentrated under reduced pressure, 50 mL of ethyl acetate was added, and stirred for 0.5 h before filtration. The filtrate was washed sequentially with 30 mL of 1M hydrochloric acid (2 times), followed by 30 mL of saturated sodium chloride, dried over sodium sulfate, concentrated, and purified by silica gel column chromatography to yield 3.6 g of a white solid. m / z: 646.10 (M+1).

[0069] Step 3:

[0070] Intermediate 2-2 (700 mg), cuprous iodide (21 mg), Pd(PPh3)Cl2 (152 mg), triethylamine (274 mg), trimethylsilylacetylene (213 mg), and 7 mL of tetrahydrofuran were added to a reaction flask. Under nitrogen, the temperature was raised to 35°C and the reaction was allowed to proceed for 2 h. The reaction mixture was passed through a celite funnel, and the filter cake was washed with 50 mL of ethyl acetate. The filtrate and washings were combined, concentrated, and purified by silica gel column chromatography to yield 589 mg of a yellow solid. m / z: 616.20 (M+1).

[0071] Step 4:

[0072] Intermediate 2-3 (589 mg) and 10 mL of acetonitrile were added to the reaction flask. After stirring at room temperature until dissolved, trimethylsilyl iodide (421 mg) was added. After stirring at room temperature for 1 hour, triethylamine (283 mg) was added. After stirring for 15 minutes, the mixture was concentrated and purified by silica gel column chromatography to obtain 1.1 g of a brown solid. m / z: 482.35 (M+1).

[0073] Step 5:

[0074] Intermediate 2-4 (1.0 g) was dissolved in 10 mL of N-methylpyrrolidone (NMP), and potassium hydroxide aqueous solution (264 mg of KOH in 10 mL of water) was added dropwise. After the addition was complete, the temperature was raised to 70°C and the reaction mixture was reacted for 3 h. After cooling to room temperature, 40 mL of water was added to the reaction system, and the mixture was extracted with 40 mL of dichloromethane twice. The organic phase was washed sequentially with 40 mL of saturated sodium chloride solution, dried over sodium sulfate, and concentrated for later use.

[0075] The concentrate from the previous step and 5 mL of DMF were added to a reaction flask. Stir at room temperature until the solution became clear. Diisopropylethylamine (DIPEA, 1.5 mL) and acryloyl chloride (20 μL) were added sequentially. After stirring at room temperature for 15 minutes, 20 mL of water was added to the reaction solution. The mixture was extracted with 20 mL x 2 of dichloromethane. The organic phase was washed with 30 mL of saturated sodium chloride, dried over sodium sulfate, concentrated, and purified on a silica gel preparative plate to yield 15 mg of an off-white solid. m / z: 310.15 (M+1). 1H NMR (DMSO-d6, 600MHz) δ = 12.05 (s, 1H), 8.20 (s, 1H), 7.56 (s, 1H), 6.83 ~ 6.78 (dd, J = 16.8Hz, J = 10.2Hz, 1H), 6.12 ~ 6.1107 (m, 2H ), 5.69~5.67(dd,J=10.8Hz,J=1.8Hz,1H),4.95~3.90(m,3H),4.27(s,1H),3.15~2.60(m,1H),1.95~1.60(m,4H),1.21(s,3H).

[0076] Example 3:

[0077] first step:

[0078] SM3-1 (4.62 g) was dissolved in DMF (46 mL). N-iodosuccinimide (NIS) (8.1 g) was added portionwise at room temperature. The mixture was heated to 50°C and stirred overnight. Upon completion, the reaction solution was slowly added dropwise to ice water, causing a large amount of solid to precipitate. The mixture was stirred for 30 min, filtered, and the filter cake was rinsed with water. The solid was collected, dried, and purified by silica gel column chromatography to yield 5.5 g of intermediate 3-1 as a yellow solid. m / z: 280.85 (M+H), 278.90 (MH).

[0079] Step 2:

[0080] Intermediate 3-1 (3 g), trimethylsilyl acetylene (6.3 g), triethylamine (1.1 g), and CuI (1 g) were weighed into a 100 mL three-necked flask. DMF (10 mL) and THF (30 mL) were added. The mixture was stirred at room temperature for 30 min under N₂ protection. Pd(PPh₃)₄ (1.24 g) was added and the mixture was allowed to react at room temperature for 3 h. The reaction solution was filtered, and the filtrate was diluted with EA, washed with water, and then with saturated NaCl. The organic phase was concentrated and purified by silica gel column chromatography to afford 2.5 g of Intermediate 3-2 as a reddish-yellow oil. m / z: 251.00 (M+H), 249.05 (MH).

[0081] Step 3:

[0082] Intermediate 3-2 (1 g), SM-2 (1.1 g), and triethylamine (980 mg) were weighed, acetonitrile (10 mL) was added, and the mixture was heated to 75°C for 3 h. After concentration, ethyl acetate (20 mL) was added to the residue, which was then washed sequentially with H2O (20 mL) and saturated NaCl (20 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to afford 1.2 g of Intermediate 3-3 as a yellow oil. m / z: 463.20 (M+H), 461.20 (MH).

[0083] Step 4:

[0084] Intermediate 3-3 (920 mg) was weighed and added to acetonitrile (10 mL). Iodotrimethylsilane (1.75 g) was added dropwise at room temperature and allowed to react for 30 min. After completion, triethylamine was added dropwise to the reaction solution to quench the reaction. The mixture was concentrated, and ethyl acetate (20 mL) and water (20 mL) were added to the residue. The mixture was separated, and the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to afford 380 mg of Intermediate 3-4 as a pale yellow solid. m / z: 329.20 (M+H), 327.15 (MH).

[0085] Step 5:

[0086] Intermediate 3-4 (220 mg) was weighed and dissolved in THF (2 mL). Aqueous LiOH solution (56 mg of LiOH dissolved in H2O (0.5 mL)) was added dropwise under ice-cooling. After completion of the dropwise addition, the mixture was stirred under ice-cooling for 30 min. After completion of the reaction, the pH was adjusted to 6-7 with 1 M HCl. EA and water were added, and the layers were separated. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to obtain 140 mg of Intermediate 3-5 as a yellow solid. m / z: 257.10 (M+H).

[0087] Step 6:

[0088] Intermediate 3-5 (140 mg) was weighed and dissolved in anhydrous DMF (1.5 mL). DIPEA (210 mg) was then added dropwise, and acryloyl chloride (44 mg) was added dropwise under ice-cooling. The mixture was stirred for 5 min. After completion of the reaction, the reaction solution was dropped into ice-cold water to quench the reaction. The mixture was extracted with dichloromethane (10 mL x 2). The organic phases were combined and washed with saturated NaCl (20 mL). The organic phases were dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to obtain 23 mg of compound 3 as a light yellow solid. m / z: 311.15 (M+H), 309.15 (MH). 1 H NMR (DMSO-d6, 600MHz) δ = 13.93 (s, 1H), 8.36 (s, 1H), 6.87 ~ 6.82 (dd, J = 16.8Hz, J = 10.8Hz, 1H), 6.61 (s, 1H), 6.15 ~ 6.13 (dd, J = 10.8Hz, J = 2.4Hz, 1H),5.74~5.72(dd,J=10.8Hz,J=2.4Hz,1H),4.74(s,1H),4.70~4.06(m ,3H),3.22~2.82(m,1H),2.05~1.68(m,4H),1.32~1.24(d,J=6.6Hz,3H).

[0089] Example 4:

[0090] first step:

[0091] Intermediate 2-2 (1.0 g) was dissolved in DMF (10 mL), followed by the addition of DIPEA (500 mg), CuI (30 mg), and Pd(PPh3)4 (179 mg). Finally, 1,7-octanediyne (329 mg) was added and the mixture was reacted at room temperature under nitrogen for 6.5 h. Ethyl acetate (15 mL) and water (25 mL) were added, and the organic layer was separated and washed with 5% LiCl solution (25 mL x 3) and saturated brine (50 mL x 1), then dried over anhydrous sodium sulfate. The filtrate was filtered, concentrated, and purified by silica gel column chromatography to yield 0.72 g of Intermediate 4-2 as an orange-red oil. m / z: 624.25 (M+H), 622.25 (MH).

[0092] Step 2:

[0093] Intermediate 4-2 (0.72 g) was dissolved in acetonitrile (7 mL), and trimethylsilyl iodide (0.36 mL) was added. The mixture was allowed to react at room temperature for 1 h. Triethylamine was added to quench the reaction, and the solvent was removed by vortexing. Ethyl acetate (14 mL) was added, and the mixture was washed with water (14 mL x 2), then dried over anhydrous sodium sulfate. The mixture was filtered, concentrated, and purified by silica gel column chromatography to obtain 230 mg of Intermediate 4-4 as an off-white solid. m / z: 490.20 (M+H), 488.20 (MH).

[0094] Step 3:

[0095] Intermediate 4-4 (230 mg) was dissolved in NMP (0.5 mL), and potassium hydroxide aqueous solution (53 mg KOH, 0.2 mL water) was added dropwise. The mixture was reacted at 70°C for 0.5 h. The temperature was cooled to room temperature, and the reaction solution was added dropwise to water (6 mL). Extraction was performed with ethyl acetate (10 mL x 1). The organic layer was washed with saturated brine (6 mL x 3), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure and subjected to silica gel column chromatography to afford 85 mg of Intermediate 4-5 as a white solid. m / z: 336.20 (M+H), 334.20 (MH).

[0096] Step 4:

[0097] Intermediate 4-5 (85 mg) was dissolved in DMF (0.8 mL), and N,N-diisopropylethylamine (98 mg) and acryloyl chloride (18 μL) were added. The mixture was allowed to react at room temperature for 15 min. The reaction solution was added dropwise to water (8 mL), extracted with dichloromethane (8 mL × 2), washed with water (8 mL × 3), washed with 5% LiCl solution (8 mL × 2), washed with saturated brine (8 mL × 1), and dried over anhydrous sodium sulfate. After filtration and silica gel concentration, column chromatography afforded 18 mg of compound 4 as a yellow solid. m / z: 390.25 (M+H), 388.20 (MH). 1H NMR (DMSO-d6, 600MHz) δ = 11.91 (s, 1H), 8.18 (s, 1H), 7.40 (s, 1H), 6.83 ~ 6.78 ( dd,J=16.2Hz,J=10.8Hz,1H),6.12~6.09(d,J=16.8Hz,1H),6.02~6.01(d,J=4 .8Hz,1H),5.70~5.68(d,J=10.2Hz,1H),4.95~3.90(m,4H),2.97(s,1H),2.77 (s,1H),2.56~2.52(m,1H),2.26~2.16(m,2H),2.06~1.55(m,8H),1.22(s,3H).

[0098] Example 5:

[0099] first step:

[0100] To Intermediate 2-2 (1.2 g) and p-phenylenevinylboronic acid (0.88 g) were added tetrahydrofuran (20 mL) and ethanol (20 mL), stirred to dissolve, and then Pd(OAc)2 (46 mg), potassium carbonate (540 mg), and water (20 mL) were added sequentially. The mixture was allowed to react at room temperature for 2 h under nitrogen. Ethyl acetate (25 mL) and water (50 mL) were added and stirred. The organic layer was separated and the aqueous layer was extracted with ethyl acetate (25 mL x 1). The combined organic layers were washed with 2M sodium hydroxide solution (50 mL x 2) and saturated sodium chloride solution (40 mL x 2). The mixture was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and purified by silica gel column chromatography to obtain 0.6 g of Intermediate 5-2 as a light yellow oil. m / z: 586.20 (M+H), 584.25 (MH).

[0101] Step 2:

[0102] Intermediate 5-2 (0.6 g) was dissolved in dichloromethane (12 mL), and trifluoroacetic acid (4 mL) was added. The mixture was allowed to react at room temperature for 0.5 h. The solvent was removed by concentration, and dichloromethane (15 mL) was added. Saturated sodium bicarbonate solution (20 mL) was added dropwise with stirring. The organic layer was separated, and the aqueous layer was extracted with dichloromethane (15 mL x 2). The combined organic layers were concentrated and purified by silica gel column chromatography to obtain 350 mg of a light yellow solid, Intermediate 5-3. m / z: 486.20 (M+H), 484.15 (MH).

[0103] Step 3:

[0104] Intermediate 5-3 (350 mg) was dissolved in NMP (1 mL), and potassium hydroxide (KOH 73 mg, water 0.45 mL) was added dropwise. The reaction was incubated at 70°C for 15 min. After cooling to room temperature, the mixture was added dropwise to 10 mL of water. A solid precipitated, which was filtered, washed with water, and air-dried to afford 160 mg of a pale yellow solid, Intermediate 5-4. m / z: 332.15 (M+H), 330.20 (MH).

[0105] Step 4:

[0106] To intermediate 5-4 (80 mg) was added tetrahydrofuran (0.8 mL) and water (0.4 mL) in sequence. After stirring and dissolving, the temperature was lowered to 15°C. Potassium phosphate (107 mg) was added. After stirring and dissolving, the temperature was lowered to below 10°C. 3-Chloropropionyl chloride (25 μL) was added dropwise and the reaction was incubated for 1 h. Sodium hydroxide solution (44 mg of sodium hydroxide and 0.5 mL of water) was added and the reaction was continued at room temperature for 4 h. The organic layer was separated and the aqueous layer was extracted with ethyl acetate (2 mL x 2). The combined organic layers were washed with saturated sodium chloride (2 mL x 1), dried over anhydrous sodium sulfate, concentrated, and purified by silica gel column chromatography to yield 26 mg of compound 5 as an off-white solid. m / z: 386.20 (M+H), 384.15 (MH). 1 H NMR (DMSO-d6, 600MHz) δ=11.96(s,1H),8.22(s,1H),7.57~7.56(d,J=7.8Hz,2H),7.53~7.52(d,J=6 .4Hz, 2H), 7.36~7.35 (d, J=2.4Hz, 1H), 6.81~5.77 (dd, J=16.8Hz, J=10.8Hz, 1H), 6.11~6.08 (dd, J=1 6.8Hz, J=1.8Hz, 1H), 5.69~5.67 (d, J=10.2Hz, 1H), 5.48~5.47 (d, J=7.8Hz, 1H), 4.90~4.10 (br, 2H), 4.22 (s, 1H), 4.06 ~ 3.95 (m, 1H), 3.06 ~ 2.55 (m, 1H), 1.90 ~ 1.52 (m, 4H), 1.14 ~ 1.13 (d,, J = 6.6Hz, 3H).

[0107] Biological testing

[0108] 1. Compound kinase JAK3 activity inhibition test

[0109] The test method is the same as that in the literature (reference: "Design of a Janus Kinase 3 (JAK3) Specific Inhibitor 1-((2S,5R)-5-((7H-Pyrrolo[2,3-d]pyrimidin-4-yl)amino)-2-methylpiperidin-1-yl)prop-2-en-1-one (PF-06651600) Allowing for the Interrogation of JAK3 Signaling in Humans", Journal Medicinal Chemistry. 2017, 60, 5, 1971-1993). That is:

[0110] Test method:

[0111] (1) Test buffer

[0112] (2) Kinase and substrate information

[0113] (3) Testing process:

[0114] Prepare 2× ATP and substrate solutions and 2× kinase and metal solutions using assay buffer;

[0115] 40 nL of compound was transferred to the 384 assay plate via Echo 655. 2 μL of 2× kinase and metal solution was added, mixed, and incubated in the 384 assay plate at 25°C for 10 min;

[0116] 2 μL of 2× substrate and ATP solution was added to the wells and incubated at 25°C for 60 min;

[0117] Add 4 μL of ADP-Glo ​​reagent to the wells and incubate at 25°C for 40 min;

[0118] Add 8 μL of kinase assay reagent to the wells and incubate at 25°C for 40 minutes;

[0119] Luminescent signals were recorded on a microtiter plate reader.

[0120] Calculation method:

[0121] The inhibition percentage is calculated as follows: Inhibition percentage = 100% - (positive control) / (negative control - positive control) × 100%

[0122] Computing IC 50 And draw the effect-dose curve of the compound:

[0123] IC50 was calculated by fitting the logarithm of % inhibition and compound concentration to a nonlinear regression (dose response - variable slope) using GraphPad 8.0. Y = Bottom + (Top - Bottom) / (1 + 10^((LogIC 50 -X)×hillslope))

[0124] X: logarithmic value of inhibition concentration; Y: percentage of inhibition.

[0125] Test results:

[0126] The inhibitory activity of the compounds of the present invention on JAK3 kinase IC 50 value

[0127] The above data show that compounds 1, 2, 3, 4, and 5 of the present invention have a good inhibitory effect on JAK3, and their JAK3 inhibitory activity is significantly better than the JAK3 inhibitory activity (IC) of compound PF-06651600 (Ritlecitinib) (numbered 11 in the literature) described in the literature (Journal Medicinal Chemistry. 2017, 60, 5, 1971-1993). 50 =33 nM).

[0128] 2. Inhibitory effect of compounds on the activity inhibition of kinase JAK family subtypes

[0129] Test principle:

[0130] The experimental purpose is to detect the inhibitory ability of the present invention on kinase activity through a drug screening system based on JAK1, JAK2, and TYK2 kinases.

[0131] Test method:

[0132] (1) Prepare 2× ATP / substrate solution and 2× kinase solution using kinase reaction buffer.

[0133] (2) Transfer 20 nL of compound dilution to a 384 assay plate using an Echo 655; after centrifugation, add 2 μL of 2× kinase and metal ion solution to the 384 assay plate, centrifuge at 1000 rpm for 1 minute, and incubate at 25°C for 10 minutes.

[0134] (3) Add 2 μL of 2× substrate and ATP solution to the 384 assay plate, centrifuge at 1000 rpm for 1 minute, and incubate at 25°C for 60 minutes.

[0135] (4) Transfer 4 μL of ADP-Glo ​​to a 384 assay plate, centrifuge at 1000 rpm for 1 minute, and incubate at 25°C for 40 minutes.

[0136] (5) Transfer 8 μL of the assay solution to a 384 assay plate, centrifuge at 1000 rpm for 1 minute, and incubate at 25°C for 40 minutes.

[0137] (6) Read the luminescence signal using a multifunctional microplate reader.

[0138] Data processing method:

[0139] The negative control reading was set as 0% inhibition rate, and the 10000nM Tofacitinib reading was set as 100% inhibition rate.

[0140] The inhibition rate of each test solution was calculated.

[0141] Average value of the ratio of 10000nM Tofacitinib control wells

[0142] Average value of negative control well ratio

[0143] The IC of the compound was obtained using the following nonlinear fitting formula: 50 (half inhibitory concentration): Y = Bottom + (Top-Bottom) / (1 + 10^((LogIC 50 -X)*HillSlope))

[0144] X: log value of compound concentration

[0145] Y: Compound inhibition rate (%inh)

[0146] Z' factor calculation equation: Z'=1-3(SDmin+SDmax) / (AVEmax-AVEmin)

[0147] in:

[0148] Min is the positive control Data value, and Max is the negative control DMSO Data value.

[0149] SD is standard error and AVE is mean.

[0150] Test results:

[0151] IC inhibitory effects of the compounds of the present invention on different kinases 50 Value (nM)

[0152] The above data show that compounds 2 and 3 of the present invention have no inhibitory effect on other subtypes of the JAK family (JAK1, JAK2, and TYK2). However, the marketed drug Tofacitinib has inhibitory effects on other subtypes of the JAK family (JAK1, JAK2, and TYK2). The above research results show that compounds 2 and 3 of the present invention have selective inhibitory effects on the JAK family kinase JAK3, but have no inhibitory effects on other subtypes of the JAK family (JAK1, JAK2, and TYK2). Marketed drugs do not have such selectivity. This shows that the compounds of the present invention have potential safety characteristics that are superior to the marketed drug Tofacitinib in future clinical use.

[0153] 3. Inhibitory effect of compounds on the expression of phosphorylated STAT5 in CD3+ cells in human PBMC

[0154] Test principle:

[0155] The phosphorylated STAT5 assay employed flow cytometry (FACS) to examine the inhibitory effect of the compounds of this invention on phosphorylated STAT5 expression in CD3+ cells of PBMCs. IL-2 activates JAK1 / JAK3 proteins in CD3+ cells, which phosphorylate STAT5. The compounds of this invention inhibit the phosphorylation function of the JAK3 protease, thereby inhibiting phosphorylated STAT5 expression. PF-06651600 (Ritlecitinib) was used as a positive reference compound.

[0156] Test method:

[0157] (1) Prepare dilution buffer and staining buffer according to the following formula:

[0158] Dilution buffer: DPBS + 0.1% BSA;

[0159] Staining buffer: DPBS + 0.2% BSA + 1mM EDTA

[0160] (2) Human peripheral blood mononuclear cells (PBMCs) were revived by adding 500,000 human PBMCs in a volume of 66.5 μL per well of a 96-well cell culture plate and incubating in a 37°C incubator for 60 minutes.

[0161] (3) Pipette 3.5 μL of the compound from the dilution plate and add it to the cell plate. Add DMSO to the control group, mix well, and incubate in a 37°C incubator for 60 minutes.

[0162] (4) IL-2 was diluted to 600 ng / mL with dilution buffer, and PE-anti-hCD3 antibody was diluted 4-fold with dilution buffer.

[0163] (5) After the 60-minute incubation, 6 μL of diluted PE-conjugated mouse anti-human CD3 antibody was added to each well, and 4 μL of diluted IL-2 was added to each well except for the ZPE control group. 4 μL of dilution buffer was added to the HPE control group, and the cells were incubated in a 37°C incubator for 30 minutes.

[0164] (6) Add 150 μL of 37°C preheated lysis fixative to each well of all cells, mix well, transfer to a 96-well deep-well plate to which 800 μL of preheated lysis fixative has been added, and incubate at 37°C in the dark for 10 minutes.

[0165] (7) After centrifugation at 600 g for 5 minutes, discard the supernatant, add 1 mL of PBS to each well, centrifuge at 600 g for 5 minutes, and wash twice.

[0166] (8) Add 400 μL of pre-chilled permeabilization buffer to the cell pellet and incubate at 4°C in the dark for 30 minutes.

[0167] (9) After centrifugation at 600 g for 5 minutes, discard the supernatant, add 1 mL of loading buffer to each well, and centrifuge at 600 g for 5 minutes to wash twice.

[0168] (10) After diluting the Alexa Fluor 647-coupled mouse anti-human phosphorylated STAT5 antibody 200-fold in staining buffer, 100 μL of the antibody dilution solution was added to each well of the cells, mixed, and incubated at 25°C in the dark for 40 minutes.

[0169] (11) Add 1 mL of loading buffer to each well, centrifuge at 600 g for 5 minutes, and wash twice.

[0170] (12) The cell pellet was resuspended in 300 μL of loading buffer and analyzed on a Beckman CytoFlex S flow cytometer.

[0171] Data Analysis:

[0172] The phosphorylated STAT5 detection experiment used FLowjo software for data processing and analysis. First, circle the cell population expressing CD3 in the PE channel on the software, and then analyze the average fluorescence intensity of the Alexa Fluor 647 channel in the CD3 cell population, which is the average fluorescence intensity of phosphorylated STAT5 expression. According to the average fluorescence intensity signal value of the ZPE control group and the HPE rate control group corresponding to each plate, the inhibition rate of phosphorylated STAT5 of each compound well is calculated using the following formula. The formula "single-well inhibition rate % = 100-100×(single-well signal value-HPE control group signal average) ÷(ZPE control group signal average-negative HPE control group signal average)" can be used to calculate the inhibition rate of each compound well. Then import the concentration and corresponding inhibition rate data into GraphPad Prism 5 software, use the Dose Response-inhibition model in the software, use the four-parameter method to fit the inhibition rate-concentration curve, and calculate the IC of the compound. 50 value.

[0173] Test results:

[0174] IC of the inhibitory effect of the compounds of the present invention on the expression of phosphorylated STAT5 in CD3+ cells in human PBMC 50 value

[0175] The above data show that the inhibitory activity of compounds 2 and 3 of the present invention on the expression of phosphorylated STAT5 in CD3+ cells in human PBMC is better than that of Ritlecitinib.

[0176] 4. Pharmacodynamic study of the compound on bovine type II collagen-induced CIA model rats

[0177] Purpose of the test:

[0178] The purpose of this experiment is to administer the test compound orally once a day for 14 consecutive days to evaluate the pharmacological effects of the test compound on the collagen-induced arthritis (CIA) model induced by bovine type II collagen in Wistar rats.

[0179] Test method:

[0180] After the acclimation period, six animals were randomly selected on day 0 as a blank control group and not immunized. The remaining 49 rats received the first immunization via intradermal injection of an emulsion prepared by mixing equal volumes of CⅡ (2 mg / mL) and CFA (4 mg / mL) at the base of the tail. A second immunization was performed using the same method on day 7. Fourteen days after the first immunization, 30 model animals were divided equally into five groups based on body weight, paw volume, and arthritis index (AI) score, plus a control group, for a total of six groups, each with six animals: control group, model group, ritlecitinib (30 mg / kg), compound 2 (3 mg / kg), compound 2 (10 mg / kg), and compound 2 (30 mg / kg). All animals were orally gavaged with either the vehicle control, positive control drug, or the test compound 2 once daily for 14 consecutive days. Bilateral hind paw volume was measured twice weekly after grouping, and the animals were weighed and the AI ​​score was calculated for all four limbs. At the end of the experimental period, 2 to 3 rats in each group were selected to take photos of their swollen toes.

[0181] Results and Analysis:

[0182] The weight of animals in the control group increased steadily during the dosing period. The weight of animals in the model group showed a trend of initially decreasing and then recovering, with the model group's weight remaining significantly lower than that of the control group throughout the experimental period. Compared to the model group, the weight gain rates of the model animals in the three dose groups of ritlecitinib (30 mg / kg) and compound 2 increased to varying degrees.

[0183] The paw volume of animals in the Model group was significantly increased from D4 to D14 compared with the Control group. The paw volume of animals in the Ritlecitinib (30 mg / kg), Compound 2 (10 mg / kg) and Compound 2 (30 mg / kg) groups was significantly decreased from D4 to D14 compared with the Model group. The paw volume of animals in the Compound 2 (3 mg / kg) group was significantly decreased from D4 to D14 compared with the Model group, but there was no statistically significant difference. The three doses of Compound 2 showed a dose-related positive correlation in reducing the paw volume of animals. The effect of the test substance on the paw volume of CIA rats induced by CⅡ after oral gavage once a day for 14 consecutive days is shown in the following table.

[0184] Among them: #P<0.05,##P<0.01vs.Control; *P<0.05,**P<0.01vs.Model.

[0185] The AI ​​scores of animals in the Model group were significantly higher than those in the Control group from D0 to D14. The AI ​​scores of animals in the Ritlecitinib (30 mg / kg), Compound 2 (10 mg / kg), and Compound 2 (30 mg / kg) groups were significantly lower from D4 to D14 compared to the Model group. The AI ​​scores of animals in the Compound 2 (3 mg / kg) group were significantly lower from D4 to D14 compared to the Model group. The three doses of Compound 2 showed a dose-dependent positive correlation in reducing the AI ​​scores of animals.

[0186] Test conclusion:

[0187] Oral gavage of Ritlecitinib (30 mg / kg) and Compound 2 (3 mg / kg, 10 mg / kg, 30 mg / kg) once a day for 14 consecutive days significantly improved the weight loss of CIA model rats, and showed different degrees of therapeutic effects on the hind limb volume and limb AI scores. The pharmacodynamic effect of the test compound 2 on CIA model rats was positively correlated with the dose, and the compound 2 (30 mg / kg) group had a better effect on alleviating the symptoms of hind limb arthritis in rats than the same dose of Ritlecitinib.

Claims

1. A compound of formula (I), or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, in, X is an oxygen atom, -NH-; Y is a nitrogen atom, -C(H)=; Z is a nitrogen atom, CR 1 ; Q is a bond, an oxygen atom, -O-CH2-, -NH-, -CH2-, -NH-CH2-, C1-C6 alkyl, C2-C8 alkynyl, C2-C8 alkynylC1-C6 alkyl, C1-C6 alkylC2-C8 alkynyl, C2-C8 alkenyl, C2-C8 alkenylC1-C6 alkyl, C1-C6 alkylC2-C8 alkenyl, aryl, C2-C8 alkynylaryl, arylC2-C8 alkynyl, or heteroaryl; R 0 , R 1 , R 3 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 Each is independently hydrogen, deuterium, halogen, alkyl, deuterated alkyl, haloalkane, cycloalkyl, heterocyclyl, heterocyclylalkyl, aryl, heteroaryl, hydroxy, amino, sulfone, phosphorus carbonyl, substituted silicon, alkenyl, alkynyl, wherein the alkyl, aryl, heteroaryl is independently optionally substituted by one or more substituents selected from the following: C1-C6 alkyl, halogen, hydroxy, methoxy, amino, cyano, alkylamino, dialkylamino, trifluoromethyl, C3-C6 cycloalkyl, C2-C8 alkynyl, or C2-C8 alkenyl; n1 = 0, 1, 2, 3, or 4; n2=0, 1, 2, 3, or 4.

2. A compound of formula (II), or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, in, Y is a nitrogen atom, -C(H)=; Z is a nitrogen atom, CR 1 ; Q is a bond, an oxygen atom, -O-CH2-, -NH-, -CH2-, -NH-CH2-, C1-C6 alkyl, C2-C8 alkynyl, C2-C8 alkynylC1-C6 alkyl, C1-C6 alkylC2-C8 alkynyl, C2-C8 alkenyl, C2-C8 alkenylC1-C6 alkyl, C1-C6 alkylC2-C8 alkenyl, aryl, C2-C8 alkynylaryl, arylC2-C8 alkynyl, or heteroaryl; R 1 , R 3 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 Each is independently hydrogen, deuterium, halogen, alkyl, deuterated alkyl, haloalkane, cycloalkyl, heterocyclyl, heterocyclylalkyl, aryl, heteroaryl, alkenyl, alkynyl, wherein the alkyl, aryl, heteroaryl are independently optionally substituted by one or more substituents selected from the following: C1-C6 alkyl, halogen, hydroxy, methoxy, amino, cyano, alkylamino, dialkylamino, trifluoromethyl, C3-C6 cycloalkyl, C2-C8 alkynyl, or C2-C8 alkenyl.

3. A compound of formula (III), or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, in, Z is a nitrogen atom, CR 1 ; R 1 , R 3 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 Each is independently hydrogen, deuterium, halogen, alkyl, deuterated alkyl, haloalkane, cycloalkyl, heterocyclyl, heterocyclylalkyl, aryl, heteroaryl, alkenyl, alkynyl, alkenylalkyl, alkynylalkyl, wherein the alkyl, aryl, heteroaryl are independently optionally substituted by one or more substituents selected from the following: C1-C6 alkyl, halogen, hydroxy, methoxy, amino, cyano, alkylamino, dialkylamino, trifluoromethyl, C3-C6 cycloalkyl, C2-C8 alkynyl, or C2-C8 alkenyl.

4. A compound of any one of the following formulae 1 to 12, or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, 5. A pharmaceutical composition comprising the compound according to any one of claims 1 to 4 or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

6. Use of the compound according to any one of claims 1 to 4 or its stereoisomer, tautomer or pharmaceutically acceptable salt or the pharmaceutical composition according to claim 5 in the preparation of a medicament for treating a disease by inhibiting Janus kinase (JAK).

7. The use according to claim 6, wherein the disease is rheumatoid arthritis, alopecia areata, vitiligo, asthma, chronic obstructive pulmonary disease, tumors, Crohn's disease, inflammatory bowel disease of ulcerative colitis, proctitis, eosinophilic gastroenteritis or mastocytosis.

8. A method for treating a disease by inhibiting Janus kinase (JAK), comprising administering a therapeutically effective dose of the compound according to any one of claims 1 to 4 or a stereoisomer, tautomer or pharmaceutically acceptable salt thereof to an individual in need thereof.

9. The method of claim 1, wherein the disease is rheumatoid arthritis, alopecia areata, vitiligo, asthma, chronic obstructive pulmonary disease, tumors, Crohn's disease, inflammatory bowel disease of ulcerative colitis, proctitis, eosinophilic gastroenteritis or mastocytosis.

10. A method for preparing the compound according to any one of claims 1 to 4, comprising the following reaction scheme: Using SM-1 as the starting material, coupling reaction with SM2 is performed to obtain compound (IM1), then coupling reaction with SM-3 is performed to obtain compound (IM2), deprotection is performed to obtain compound (IM-3), and finally coupling reaction with SM-4 is performed to obtain compound II; in, Z, Q, R 3 , R 8 , R 9 , R 10 , R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 Definition as in claim 1.