Compound with TYK2 inhibiting effect as well as preparation method and application thereof

Abstract

The invention provides a compound with a TYK2 inhibiting effect as well as a preparation method and application thereof, and belongs to the technical field of medicinal chemistry. The structure of the compound is shown as a formula I in the specification. The compound provided by the invention shows good binding activity to TYK2JH2, and can effectively inhibit a TYK2 signal channel stimulated and activated by IFN-alpha, IL-12 and IL-23 in human PBMC (peripheral blood mononuclear cells); the compound disclosed by the invention is excellent in physicochemical property, solubility and permeability, has remarkable advantages in preparation development and production and clinical application, and has a wide application prospect in preparation of drugs for preventing and / or treating diseases related to TYK2 activity.

Description

A compound with TYK2 inhibitory effect and preparation method and use thereof TECHNICAL FIELD

[0001] The present application belongs to the technical field of pharmaceutical chemistry, and particularly relates to a compound with TYK2 inhibitory effect and preparation method and use thereof. BACKGROUND

[0002] TYK2 is one of the members of the Janus kinase family (JAK). TYK2 plays a key role in mediating the signal transduction of a variety of cytokines, especially type I interferons, interleukins IL-12 and IL-23, etc., which are related to inflammation and immune response. TYK2 inhibitors can strongly and specifically inhibit the activation of TYK2 by binding to the protein domain of TYK2, while not affecting the function of other JAK family targets. This selective inhibition can reduce side effects and improve the safety and tolerability of treatment.

[0003] TYK2 inhibitors can be used for the treatment of a variety of inflammatory and autoimmune diseases. For example, the first TYK2 allosteric inhibitor in the world, deucalixibatin (trade name: Sondium), selectively targets TYK2 through a unique "allosteric inhibition" mechanism, thereby inhibiting the signal transduction of interleukin (IL)-23, IL-12 and type I interferon (IFN), which are key cytokines involved in the pathogenesis of psoriasis, and precisely targets the therapeutic effect while bringing good safety, providing a new oral targeted treatment option for psoriasis patients. In addition, TYK2 inhibitors have entered the clinical research stage for systemic lupus erythematosus, ulcerative colitis, Crohn's disease and other indications.

[0004] As known to those skilled in the art, good solubility of small molecule drugs is an important advantage in drug development and application. It not only helps to improve the bioavailability, stability and safety of the drug, but also simplifies the formulation development and production process, reduces production costs and improves patient compliance. Chinese Patent Application No. CN111194317A discloses a compound that can be used to inhibit TYK2, for example However, the solubility of the compound is poor, which limits its formulation development and clinical application. SUMMARY

[0005] The present application aims to provide a compound with TYK2 inhibitory effect and preparation method and use thereof.

[0006] The present application provides a compound, a pharmaceutically acceptable salt thereof, a stereoisomer thereof, a deuterated compound thereof or a solvate thereof, the structure of the compound being shown in formula I:

[0007] wherein, is a 5-membered heteroaromatic ring containing two nitrogen atoms, E1 is selected from CH or N, E2 is selected from CH or N, E3 is selected from CH or N, E4 is selected from CH or N;

[0008] X is selected from O, S, NH, C 1-6 alkylene, C 2-6 alkenylene, C 2-6 alkynylene, 3-8 membered saturated cycloalkyl, 3-8 membered saturated heterocyclyl;

[0009] Y is selected from CONH, NHCO, CO, COO, OCO, NH;

[0010] is selected from =0, =S, =CH2, -H, -R 1m , NR 1c R 1d , CONR 1c R 1d , COOR 1e ;

[0011] R 1m is selected from C 1-8 alkyl, C 1-8 alkoxy, halogen;

[0012] R 1c is selected from hydrogen, C 1-8 alkyl, R 1d is selected from hydrogen, C 1-8 alkyl, COR 1f , unsubstituted or substituted by one or more than one R 1g 4-8 membered heteroaromatic ring, 4-8 membered aromatic ring, 3-8 membered saturated cycloalkyl, 3-8 membered saturated heterocyclyl; or, R 1c , R 1d are linked to form a 3-8 membered saturated heterocyclyl;

[0013] R 1e is selected from C 1-8 alkyl, R 1f is selected from C 1-8 alkyl, R 1g is selected from C 1-8 alkyl;

[0014] m is selected from 0, 1, 2 or 3;

[0015] n is selected from 0, 1, 2, 3 or 4;

[0016] R2is each independently selected from hydrogen, C 1-8 alkyl, C 1-8 alkoxy, halogen;

[0017] Z is N;

[0018] A ring is absent or is selected from a 4-8 membered heteroaromatic ring, a 4-8 membered aromatic ring, a 3-8 membered saturated cycloalkyl, a 3-8 membered saturated heterocyclyl;

[0019] a is selected from 0, 1, 2, 3 or 4;

[0020] R3is each independently selected from hydrogen, halogen, COOR 3a , OCOR 3a , COR 3a , OR 3a , unsubstituted or substituted by one or more than two R 3b , C 1-8 alkyl, C 2-8 alkenyl, C 2-8 alkynyl, 3-8 membered saturated cycloalkyl, 3-8 membered saturated heterocyclyl; R 3a is selected from C 1-8 alkyl, R 3b is each independently selected from halogen, C 1-8 alkyl;

[0021] B ring is selected from a 3-8 membered saturated cycloalkyl, a 3-8 membered saturated heterocyclyl;

[0022] b is selected from 0, 1, 2, 3 or 4;

[0023] R4is each independently selected from hydrogen, halogen, COOR 4a , OCOR 4a , COR 4a , OR 4a , unsubstituted or substituted by one or more than two R 4b , C 1-8 alkyl, C 2-8 alkenyl, C 2-8 alkynyl, 3-8 membered saturated cycloalkyl, 3-8 membered saturated nitrogen containing heterocyclyl; R 4a is selected from C 1-8 alkyl, R 4b is each independently selected from halogen, C 1-8 alkyl;

[0024] R5is selected from C 1-8 alkyl.

[0025] Further, the structure of the compound is shown as Formula II:

[0026] wherein, is a 5-membered heteroaromatic ring containing two nitrogen atoms, E1is selected from CH or N, E2is selected from CH or N, E3is selected from CH or N, E4is selected from CH or N;

[0027] X is selected from O, S, NH, C 1-6 alkylene, C 2-6 alkenylene, C 2-6 alkynylene, 3-8 membered saturated cycloalkyl, 3-8 membered saturated heterocyclyl;

[0028] Y is selected from CONH, NHCO, CO, COO, OCO, NH;

[0029] selected from =0, =S, =CH2, -H, -R 1m , NR 1c R 1d , CONR 1c R 1d , COOR 1e ;

[0030] R 1m is selected from C 1-8 alkyl, C 1-8 alkoxy, halogen;

[0031] R 1c is selected from hydrogen, C 1-8 alkyl, R 1d is selected from hydrogen, C 1-8 alkyl, COR 1f , unsubstituted or substituted by one or more R 1g 5-6 membered heteroaromatic ring, benzene ring, 5-6 membered saturated cycloalkyl, 5-6 membered saturated heterocyclyl; or, R 1c , R 1d are linked to form a 5-6 membered saturated heterocyclyl;

[0032] R 1e is selected from C 1-8 alkyl, R 1f is selected from C 1-8 alkyl, R 1g is selected from C 1-8 alkyl;

[0033] m is selected from 0, 1, 2 or 3;

[0034] n is selected from 0, 1, 2, 3 or 4;

[0035] R2is each independently selected from hydrogen, C 1-8 alkyl, C 1-8 alkoxy, halogen;

[0036] Z is N;

[0037] A ring is absent, or is selected from 4-8 membered heteroaromatic ring, 4-8 membered aromatic ring, 3-8 membered saturated cycloalkyl, 3-8 membered saturated heterocyclyl;

[0038] a is selected from 0, 1, 2, 3 or 4;

[0039] R3is each independently selected from hydrogen, halogen, COOR 3a , OCOR 3a , COR 3a , OR 3a , unsubstituted or substituted by one or more than two R 3b ; C 1-8 alkyl, C 2-8 alkenyl, C 2-8 alkynyl, 3-8 membered saturated cycloalkyl, 3-8 membered saturated heterocyclyl; R 3a is selected from C 1-8 alkyl, R 3b is each independently selected from halogen, C 1-8 alkyl;

[0040] B ring is selected from 3-8 membered saturated cycloalkyl, 3-8 membered saturated heterocyclyl;

[0041] b is selected from 0, 1, 2, 3 or 4;

[0042] R4is each independently selected from hydrogen, halogen, COOR 4a , OCOR 4a , COR 4a , OR 4a , unsubstituted or substituted by one or more than two R 4b ; C 1-8 alkyl, C 2-8 alkenyl, C 2-8 alkynyl, 3-8 membered saturated cycloalkyl, 3-8 membered saturated nitrogen containing heterocyclyl; R 4a is selected from C 1-8 alkyl, R 4b is each independently selected from halogen, C 1-8 alkyl;

[0043] R5is selected from C 1-8 alkyl.

[0044] Further, the structure of the compound is shown in Formula III-1, Formula III-1 or Formula III-3:

[0045] Further, the is selected from =O, =S, =CH2, -H, -R 1m , -NR 1c R 1d , -CONR 1c R 1d , -COOR 1e ;

[0046] R 1m selected from C 1-6 alkyl;

[0047] R 1c selected from hydrogen, C 1-6 alkyl, R 1d selected from hydrogen, C 1-6 alkyl, COR 1f , unsubstituted or substituted by one or more R 1g 5-6 membered heteroaromatic ring, benzene ring, 5-6 membered saturated cycloalkyl, 5-6 membered saturated heterocyclyl; or, R 1c , R 1d form a 5-6 membered saturated heterocyclyl ring;

[0048] R 1e selected from C 1-6 alkyl; 1f selected from C 1-6 alkyl; 1g selected from C 1-6 alkyl;

[0049] X is selected from O, S, NH, C 1-6 alkylene, C 2-6 alkenylene, C 2-6 alkynylene, 3-6 membered saturated cycloalkyl, 3-6 membered saturated heterocyclyl;

[0050] Y is selected from CONH, NHCO, CO, COO, OCO, NH.

[0051] Further, the structure of the compound is shown in Formula IV-1, Formula IV-2, or Formula IV-3:

[0052] wherein R 1m is selected from C 1-6 alkyl;

[0053] X is selected from O, S, NH, CH2, HC=CH, C≡C, 3-6 membered saturated cycloalkyl;

[0054] m is selected from 0, 1 or 2;

[0055] R2is selected from hydrogen, C 1-6 alkyl;

[0056] A ring is selected from 5-6 membered heteroaromatic ring, benzene ring, 3-6 membered saturated cycloalkyl, 3-6 membered saturated heterocyclyl;

[0057] a is selected from 0, 1, 2, 3 or 4;

[0058] R3is each independently selected from hydrogen, halogen, COOR3a OCOR 3a COR 3a OR 3a Not replaced or replaced by one or more R 3b The following groups are substituted: C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 Alkyne group, 3-6 membered saturated cycloalkyl group, 3-6 membered saturated heterocyclic group; R 3a Selected from C 1-6 Alkyl, R 3b Each is independently selected from halogens, C 1-6 alkyl;

[0059] Ring B is selected from 3-6 membered saturated cycloalkyl groups and 3-6 membered saturated heterocyclic groups;

[0060] b is selected from 0, 1, 2, 3, or 4;

[0061] R4 is independently selected from hydrogen, halogens, and COOR. 4a OCOR 4a COR 4a OR 4a Not replaced or replaced by one or more R 4b The following groups are substituted: C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 Alkyne group, 3-6 membered saturated cycloalkyl group, 3-6 membered saturated nitrogen-containing heterocyclic group; R 4a Selected from C 1-6 Alkyl, R 4b Each is independently selected from halogens, C 1-6 alkyl;

[0062] R5 is selected from C 1-6 alkyl.

[0063] Furthermore, the structure of the compound is shown in formula V-1, formula V-2, or formula V-3:

[0064] Among them, R 1m Selected from C 1-6 alkyl;

[0065] X is selected from O, S, NH, CH2, HC=CHH, C≡C, and 3-6 membered saturated cycloalkyl groups;

[0066] m is selected from 0, 1, or 2;

[0067] R2 is selected from hydrogen, C 1-6 alkyl;

[0068] A ring is selected from a 5-6 membered heteroaromatic ring, a benzene ring, a 3-6 membered saturated cycloalkyl, a 3-6 membered saturated heterocyclyl;

[0069] a is selected from 0, 1, 2, 3 or 4;

[0070] R3is each independently selected from hydrogen, halogen, COOR 3a , OCOR 3a , COR 3a , OR 3a , the following group which is unsubstituted or substituted with one or two or more R 3b substituents: C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, 3-6 membered saturated cycloalkyl, 3-6 membered saturated heterocyclyl; R 3a is selected from C 1-6 alkyl, R 3b is each independently selected from halogen, C 1-6 alkyl;

[0071] B ring is selected from a 3-6 membered saturated cycloalkyl, a 3-6 membered saturated heterocyclyl;

[0072] b is selected from 0, 1, 2, 3 or 4;

[0073] R4is each independently selected from hydrogen, halogen, COOR 4a , OCOR 4a , COR 4a , OR 4a , the following group which is unsubstituted or substituted with one or two or more R 4b substituents: C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, 3-6 membered saturated cycloalkyl, 3-6 membered saturated nitrogen-containing heterocyclyl; R 4a is selected from C 1-6 alkyl, R 4b is each independently selected from halogen, C 1-6 alkyl;

[0074] R5is selected from C 1-6 alkyl.

[0075] Further, the structure of the compound is shown in Formula VI-1, Formula VI-2 or Formula VI-3:

[0076] wherein R 1m is selected from C 1-6 alkyl;

[0077] X is selected from O, S, NH, CH2, HC=CH, C≡C, 3-6 membered saturated cycloalkyl;

[0078] m is selected from 0, 1 or 2;

[0079] R2is selected from hydrogen, C 1-6 alkyl;

[0080] A ring is selected from a 5-6 membered heteroaromatic ring, a benzene ring, a 3-6 membered saturated cycloalkyl, a 3-6 membered saturated heterocyclyl;

[0081] a is selected from 0, 1, 2, 3 or 4;

[0082] R3is each independently selected from hydrogen, halogen, COOR 3a , OCOR 3a , COR 3a , OR 3a , unsubstituted or substituted by one or more than one R 3b C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, 3-6 membered saturated cycloalkyl, 3-6 membered saturated heterocyclyl; R 3a is selected from C 1-6 alkyl, R 3b is each independently selected from halogen, C 1-6 alkyl;

[0083] B ring is selected from a 3-6 membered saturated cycloalkyl, a 3-6 membered saturated heterocyclyl;

[0084] b is selected from 0, 1, 2, 3 or 4;

[0085] R4is each independently selected from hydrogen, halogen, COOR 4a , OCOR 4a , COR 4a , OR 4a , unsubstituted or substituted by one or more than one R 4b C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, 3-6 membered saturated cycloalkyl, 3-6 membered saturated nitrogen containing heterocyclyl; R 4a is selected from C 1-6 alkyl, R 4b is each independently selected from halogen, C 1-6 alkyl;

[0086] R5is selected from C 1-6 alkyl.

[0087] Further, said R 1m is selected from C 1-3 alkyl;

[0088] X is selected from O, S, NH, CH2, HC=CHH, C≡C, cyclopropyl;

[0089] m is selected from 0, 1 or 2;

[0090] R2is selected from hydrogen, C 1-3 alkyl;

[0091] A ring is selected from a benzene ring;

[0092] a is selected from 0, 1, 2, 3 or 4;

[0093] R3is each independently selected from hydrogen, halogen, COOR 3a , OCOR 3a , COR 3a , OR 3a , unsubstituted or substituted by one or more than two R 3b , C 1-3 alkyl, C 2-3 alkenyl, C 2-3 alkynyl, 3-6 membered saturated cycloalkyl, 3-6 membered saturated nitrogen containing heterocyclyl; R 3a is selected from C 1-3 alkyl, R 3b is each independently selected from halogen, C 1-3 alkyl.

[0094] Further, in formula VII, the 5-6 membered nitrogen heteroaromatic ring is selected from

[0095] Further, B ring is selected from 3-6 membered saturated cycloalkyl, 3-6 membered saturated heterocyclyl;

[0096] b is selected from 0, 1, 2, 3 or 4;

[0097] R4is each independently selected from hydrogen, halogen, COOR 4a , OCOR 4a , COR 4a , OR 4a , unsubstituted or substituted by one or more than two R 4b , C 1-3 alkyl, C 2-3 alkenyl, C 2-3 alkynyl, 3-6 membered saturated cycloalkyl, 3-6 membered saturated nitrogen containing heterocyclyl; R 4a is selected from C 1-3 alkyl, R 4b is each independently selected from halogen, C 1-3 alkyl;

[0098] R5is selected from C 1-3 alkyl.

[0099] Further, the B ring is selected from 3-membered saturated cycloalkyl, 4-membered saturated cycloalkyl, 5-membered saturated cycloalkyl, 6-membered saturated cycloalkyl.

[0100] Further, the For

[0101] Further, the compound is one of the following compounds:

[0102] The present application also provides a pharmaceutical composition, which is a preparation prepared from the above-mentioned compound, pharmaceutically acceptable salt thereof, stereoisomer thereof, deuterated compound thereof or solvate thereof as an active ingredient, and a pharmaceutically acceptable adjuvant.

[0103] The present application also provides the use of the above-mentioned compound, pharmaceutically acceptable salt thereof, stereoisomer thereof, deuterated compound thereof or solvate thereof in the preparation of a TYK2 inhibitor.

[0104] Further, the TYK2 inhibitor is a drug for preventing and / or treating inflammation, skin disease, autoimmune disease, cancer or neurodegenerative disease.

[0105] Further, the autoimmune disease is lupus, rheumatoid arthritis, multiple sclerosis, psoriasis, ulcerative colitis, Crohn's disease, type I diabetes or Sjogren's syndrome; the skin disease is vitiligo, skin rash, atopic dermatitis or lichen planus; the cancer is kidney cancer, liver cancer, pancreatic cancer, gastric cancer, breast cancer, prostate cancer, head and neck cancer, thyroid cancer, lung cancer, glioblastoma, melanoma, lymphoma or leukemia; the neurodegenerative disease is Alzheimer's disease, Parkinson's disease, Huntington's disease or amyotrophic lateral sclerosis.

[0106] The compound provided by the present application exhibits good binding activity to TYK2 JH2 and only has weak or no inhibitory activity to other JAK kinases; in human PBMC cells, it can effectively inhibit the TYK2 signal pathway stimulated and activated by IFN-alpha, IL-12 and IL-23, and can be used for preparing a TYK2 inhibitor.

[0107] The compound provided by the present application has excellent physical and chemical properties, good permeability and excellent solubility, and has significant advantages in formulation development and production and clinical application, and has a broad application prospect in the preparation of drugs for preventing and / or treating diseases related to TYK2 activity.

[0108] Obviously, according to the above content of the present application, other various forms of modification, replacement or change can be made according to the common technical knowledge and usual means in the art without departing from the above basic technical idea of the present application.

[0109] The above content of the present application will be further explained in detail by a specific embodiment in the form of examples. However, it should not be understood that the above subject matter of the present application is limited to the following examples. Any technology realized based on the above content of the present application belongs to the scope of the present application. DETAILED DESCRIPTION

[0110] The raw materials and equipment used in the present application are known products, which are obtained by purchasing commercially available products.

[0111] Example: Synthesis of the target compound of the present application

[0112] Synthesis of compound 1

[0113] N-[(1R,2R)-2-methoxycyclobutyl]-7-(methylamino)-5-{[2-oxo-1-(pyridin-2-yl)hexahydropyridin-3-yl]amino}pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0114] Step 1:

[0115] Preparation of [(2-oxohexahydropyridin-3-yl)amino]methane acid-2-methylprop-2-yl ester

[0116] A solution of 3-aminopiperidin-2-one (1000 mg, 8.760 mmol) in DCM was added to Boc2O (2007 mg, 9.198 mmol) and Et3N (930 mg, 9.198 mmol). The reaction mixture was stirred at 25°C for 16 hours, the reaction solution was concentrated under reduced pressure, and the product (1100 mg, yield 52%) was obtained as a white solid by column chromatography (DCM:MeOH=98:2). LC / MS: 215.0 [M+H] + .

[0117] Step 2:

[0118] {[2-oxo-1-(pyridin-2-yl)hexahydropyridin-3-yl]amino}methane acid-2-methylprop-2-yl ester

[0119] To a solution of [(2-oxopiperidin-3-yl)amino]methane acid 2-methylprop-2-yl ester (1100 mg, 5.109 mmol) in dioxane was added 2-iodopyridine (2095 mg, 10.219 mmol), Cul (194 mg, 1.0219 mmol), N,N-dimethyl-1,2-ethanediamine (180 mg, 2.043 mmol) and K2CO3 (2118 mg, 15.329 mmol). The reaction mixture was stirred at 100 °C for 16 h, the reaction was diluted with water (200 mL) and extracted with EA (100 mL x 3). The organic phases were combined, dried over anhydrous Na2SO4, filtered and the solution was concentrated. Purification by column chromatography (DCM:MeOH = 95:5) gave the product (550 mg, 35% yield) as a white solid. LC / MS: 291.9 [M+H] + .

[0120] Step 3:

[0121] Preparation of 3-amino-1-(pyridin-2-yl)piperidin-2-one

[0122] To a solution of {[2-oxo-1-(pyridin-2-yl)piperidin-3-yl]amino}methane acid 2- methylprop-2-yl ester (550 mg, 1.881 mmol) in DCM and trifluoroacetic acid, the reaction mixture was stirred at 25 °C for 2 h. The reaction was concentrated, diluted with a saturated NaHCO3solution (200 mL) and extracted with DCM (100 mL x 3). The organic phases were combined, dried over anhydrous Na2SO4, filtered and the solution was concentrated to give the product (300 mg, 75% yield) as a yellow solid. LC / MS: 191.9 [M+H] + .

[0123] Step 4:

[0124] Preparation of 5-chloro-7-(methylamino)pyrazolo[1,5-a]pyrimidine-3-carboxylic acid

[0125] To a solution of ethyl 5-chloro-7-(methylamino)pyrazolo[1,5-a]pyrimidine-3- carboxylate (900 mg, 3.534 mmol) in 1,4-dioxane was added a solution of NaOH (353 mg, 8.835 mmol) in water, the reaction mixture was stirred at 50 °C for 16 h. The reaction was concentrated and diluted with water, the pH of the system was adjusted to 5 with HC1, then filtered and concentrated under reduced pressure to give the product (750 mg, 84% yield) as a white solid. LC / MS: 227.1 [M+H] + .

[0126] Step 5:

[0127] Preparation of 5-chloro-N-[(1R,2R)-2-methoxycyclobutyl]-7- (methylamino)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0128] To a solution of 5-chloro-7-(methylamino)pyrazolo[1,5-a]pyrimidine-3- carboxylic acid (750 mg, 3.309 mmol) in DCM was added HATU (2517 mg, 6.619 mmol), TEA (1674 mg, 16.548 mmol) and (1R,2R)-2-methoxycyclobutyl-1- amine (402 mg, 3.971 mmol). The reaction mixture was stirred at 25 °C for 2 h. The reaction was diluted with H2O (200 mL) and extracted with DCM (100 mL x 3). The combined organic layers were dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (DCM:MeOH = 98:2) to give the product (900 mg, 79% yield) as a white solid. LC / MS: 309.8 [M+H] +

[0129] Step 6:

[0130] Preparation of {[5-chloro-3-({[(1R,2R)-2-methoxycyclobutyl]amino}carbonyl)pyrazolo[1,5- a]pyrimidin-7-yl](methyl)amino}methane acid-2-methylprop-2-yl ester

[0131] To a solution of 5-chloro-N-[(1R,2R)-2-methoxycyclobutyl]-7-(methylamino)pyrazolo[1,5- a]pyrimidine-3-carboxamide (900 mg, 2.905 mmol) in DCM was added di-tert-butyl dicarbonate (1268 mg, 5.811 mmol), triethylamine (294 mg, 2.905 mmol) and DMAP (35 mg, 0.290 mmol). The reaction mixture was stirred at 25 °C for 16 h, diluted with water and extracted with DCM (100 mL x 3). The combined organic layers were dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (DCM:EA = 4:1) to give the product (500 mg, 37% yield) as a yellow solid. LC / MS: 410.1 [M+H] + .

[0132] Step 7:

[0133] Preparation of {[3-({[(1R,2R)-2-methoxycyclobutyl]amino}carbonyl)-5-{[2-oxo-1-(pyridin-2- yl)hexahydropyridin-3-yl]amino}pyrazolo[1,5-a]pyrimidin-7-yl](methyl)amino}methane acid-2- methylprop-2-yl ester

[0134] To a solution of { [5-chloro-3-({ [(1R,2R)-2-methoxycyclobutyl] amino} carbonyl) pyrazolo[1,5-a]pyrimidin-7-yl] (methyl) amino}methane acid-2-methylprop-2-yl ester (300 mg, 0.730 mmol) in 1,4-dioxane was added 3-amino-1-(pyridin-2-yl)piperidin-2-one (167 mg, 0.876 mmol), Pd2(dba)3(67 mg, 0.073 mmol), Xantphos (42 mg, 0.073 mmol) and Cs2CO3(713 mg, 2.190 mmol) under nitrogen. The reaction mixture was stirred at 80 °C for 2 h, diluted with H2O (100 mL) and extracted with EA (50 mL x 3). The combined organic layers were dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (CH2Cl2 / MeOH = 92:8) to give the product (90 mg, 13% yield) as a yellow solid. LC / MS: 564.8 [M+H] +

[0135] Step 8:

[0136] Preparation of N-[(1R,2R)-2-methoxycyclobutyl]-7-(methylamino)-5-{[2-oxo-1-(pyridin-2- yl)hexahydropyridin-3-yl]amino}pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0137] { [3-({ [(1R,2R)-2-methoxycyclobutyl] amino} carbonyl)-5-{ [2-oxo-1-(pyridin-2- yl)hexahydropyridin-3-yl] amino} pyrazolo[1,5-a]pyrimidin-7-yl] (methyl) amino}methane acid-2-methylprop-2-yl ester (90 mg, 0.159 mmol) was added into a mixture of DCM / CF3COOH, the reaction mixture was stirred at 25 °C for 2 h. The reaction was concentrated and purified by preparative HPLC to give compound 1 (23 mg, 29.8% yield) as a white solid. LC / MS: 464.8 [M+H] + .

[0138] 1H NMR (400 MHz, CDC13) δ 8.47-8.44 (m, 1H), 8.29-8.24 (m, 2H), 7.88-7.84 (m, 1H), 7.76-7.71 (m, 1H), 7.17-7.13 (m, 1H), 6.17 (s, 1H), 6.11 (s, 1H), 5.25 (s, 1H), 4.83-4.76 (m, 1H), 4.62-4.47 (m, 2H), 3.89-3.70 (m, 2H), 3.36 (d, J = 10.4 Hz, 3H), 3.04-2.86 (m, 4H), 2.35-2.23 (m, 1H), 2.23-2.03 (m, 3H), 1.84-1.61 (m, 2H), 1.48-1.37 (m, 1H).

[0139] Step 9:

[0140] Preparation of compounds 1-P1 and 1-P2

[0141] Compound 1 was further purified by SFC separation (Chromatographic column: chiralpak-OJ, mobile phase: CO2-MEOH (DEA)) to give peak 1, enantiomer 1 (1-P1), 5-{[(3R)-2-oxo-1-(pyridin-2-yl)hexahydropyridin-3-yl]amino}-N-[(1R,2R)-2- methoxycyclobutyl]-7-(methylamino)pyrazolo[1,5-a]pyrimidine-3-carboxamide or 5-{[(3S)-2-oxo-1-(pyridin-2-yl)hexahydropyridin-3-yl]amino}-N-[(1R,2R)-2- methoxycyclobutyl]-7-(methylamino)pyrazolo[1,5-a]pyrimidine-3-carboxamide. A single configuration white solid.

[0142] LC / MS: 464.8 [M+H] + . 1H NMR (400 MHz, DMSO) δ 8.40 (ddd, J = 4.9, 1.9, 0.9 Hz, 1H), 8.26 (d, J = 9.2 Hz, 1H), 8.03 (s, 1H), 7.87 - 7.73 (m, 2H), 7.68 - 7.49 (m, 2H), 7.16 (ddd, J = 7.1, 4.9, 1.2 Hz, 1H), 5.45 (s, 1H), 4.95 (d, J = 4.9 Hz, 1H), 4.60 - 4.35 (m, 1H), 4.29 - 4.05 (m, 1H), 3.78-3.75 (m, 1H), 3.47 - 3.37 (m, 1H), 2.97 (s, 3H), 2.83 (d, J = 4.9 Hz, 3H), 2.44 - 2.32 (m, 1H), 2.14 - 1.89 (m, 3H), 1.80-1.75 (m, 2H), 1.37-1.30 (m, 2H).

[0143] Peak 2, Enantiomer 2 (1-P2), 5-{[(3S)-2-oxo-1-(pyridin-2-yl)hexahydropyridin-3- yl]amino}-N-[(1R,2R)-2-methoxycyclobutyl]-7-(methylamino)pyrazolo[1,5-a]pyrimidine- 3-carboxamide or 5-{[(3R)-2-oxo-1-(pyridin-2-yl)hexahydropyridin-3-yl]amino}-N-[(1R,2R)- 2-methoxycyclobutyl]-7-(methylamino)pyrazolo[1,5-a]pyrimidine-3-carboxamide. A single configuration white solid.

[0144] LC / MS: 464.8 [M+H] + . 1 H NMR (400 MHz, DMSO) δ 8.44 - 8.36 (m, 1H), 8.26 (d, J = 8.5 Hz, 1H), 8.08 (s, 1H), 7.81 - 7.76 (m, 2H), 7.68 - 7.56 (m, 2H), 7.17-7.14 (m, 1H), 5.46 (s, 1H), 4.94 (s, 1H), 4.39 (d, J = 11.6 Hz, 1H), 4.29 - 3.95 (m, 1H), 3.79 (ddd, J = 12.5, 7.6, 4.7 Hz, 1H), 3.66 (q, J = 7.6 Hz, 1H), 3.15 (s, 3H), 2.83 (d, J = 4.7 Hz, 3H), 2.44 - 2.40 (m, 1H), 2.13 - 1.73 (m, 6H), 1.41 - 1.29 (m, 1H).

[0145] Synthesis of Compound 2

[0146] N-[(1R,2R)-2-methoxycyclobutyl]-7-(methylamino)-5-[(2-oxo-1-phenylhexahydropyridin-3- yl)amino]pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0147] Step 1:

[0148] Preparation of {[3-({[(1R,2R)-2-methoxycyclobutyl]amino}carbonyl)-5-[(2-oxo-1- phenylhexahydropyridin-3-yl)amino]pyrazolo[1,5-a]pyrimidin-7-yl}(methyl)amino}methane acid- 2-methylprop-2-yl ester

[0149] {[5-chloro-3-({[(1R,2R)-2-methoxycyclobutyl]amino}carbonyl)pyrazolo[1,5-a]pyrimidin-7- yl](methyl)amino}methane acid-2-methylprop-2-yl ester (150 mg, 0.37 mmol), 3-amino-1- phenylpiperidin-2-one (104.19 mg, 0.55 mmol), PD2(DBA)3(66.87 mg, 0.07 mmol), Xantphos (42.25 mg, 0.07 mmol), Cs2CO3(356.87 mg, 1.09 mmol) were mixed in dioxane, the reaction mixture was stirred at 90 °C for 7 h. Diluted with water and extracted with DCM (20 mL x 3), the combined organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure. Purified by column chromatography (CH2Cl2 / MeOH = 96:4) to give the product (50 mg, yield 23%) as a white solid. LC / MS: 563.8 [M+H]+.

[0150] Step 2:

[0151] Preparation of N-[(1R,2R)-2-methoxycyclobutyl]-7-(methylamino)-5-[(2-oxo-1- phenylhexahydropyridin-3-yl)amino]pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0152] {[3-({[(1R,2R)-2-methoxycyclobutyl]amino}carbonyl)-5-[(2-oxo-1- phenylhexahydropyridin-3-yl)amino]pyrazolo[1,5-a]pyrimidin-7-yl}(methyl)amino}methane acid- 2-methylprop-2-yl ester (50 mg, 0.09 mmol) was dissolved in trifluoroacetic acid / DCM mixture, stirred at room temperature for 1 h. Purified by preparative HPLC after concentration under reduced pressure to give Cmpd 2 (28 mg, yield 68%) as a white solid. LC / MS: 463.8 [M+H] + .

[0153] 1 H NMR (400 MHz, MeOD) d = 8.24 (s, 1H), 7.48-7.44 (m, 2H), 7.36-7.30 (m, 2H), 5.58 (s, 1H), 4.342-4.36 (m, 1H), 3.89-3.80 (m, 4H), 3.36 (s, 3H), 3.08 (s, 3H), 2.60-2.52 (m, 1H), 2.30-2.18 (m, 2H), 2.18-2.11 (m, 2H), 2.10-2.00 (m, 1H), 1.70-1.46 (m, 2H).

[0154] Synthesis of compounds 3-P1 and 3-P2

[0155] 5-{[(3S)-2-oxo-1-(pyridin-2-yl)hexahydropyridin-3-yl]oxy}-N-[(1R,2R)-2- methoxycyclobutyl]-7-(methylamino)pyrazolo[1,5-a]pyrimidine-3-carboxamide or 5-{[(3R)-2-oxo-1-(pyridin-2-yl)hexahydropyridin-3-yl]oxy}-N-[(1R,2R)-2- methoxycyclobutyl]-7-(methylamino)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0156] Step 1 :

[0157] Preparation of 3-{[dimethyl(2-methylpropan-2-yl)silyl]oxy}hexahydropyridin-2-one

[0158] 3-Hydroxypiperidin-2-one (1.00 g, 8.686 mmol), imidazole (0.65 g, 9.554 mmol) and TBSC1 (1.70 g, 11.292 mmol) were dissolved in 10 mL of DMF and stirred at 25 °C for 16 hours. The reaction mixture was diluted with 50 mL of water and extracted with 30 mL of ethyl acetate (EA) three times. The organic layers were combined, dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (PE:EA = 1 :1) to give the product as a white solid (1.50 g, yield 67%). LC / MS: 230.0 [M+H] + .

[0159] Step 2:

[0160] Preparation of 3-{[dimethyl(2-methylpropan-2-yl)silyl]oxy}-1-(pyridin-2-yl)hexahydropyridin-2-one

[0161] 3-{[dimethyl(2-methylprop-2-yl)silyl]oxy}hexahydropyridin-2-one (500 mg, 2.180 mmol) was dissolved in 1,4-dioxane (10 mL), followed by the addition of 2-iodopyridine (894 mg, 4.359 mmol). The reaction mixture was degassed for 10 min under argon protection, and then potassium carbonate (904 mg, 6.539 mmol), N,N-dimethylethylenediamine (77 mg, 0.872 mmol), and copper iodide (83 mg, 0.436 mmol) were added. The reaction mixture was heated to 110 °C and reacted for 16 h. The reaction mixture was filtered and extracted three times with ethyl acetate (EA). The organic layers were combined, dried over sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (PE:EA = 9:1) to give a yellow oily product (400 mg, 56% yield). LC / MS: 306.9 [M+H] + .

[0162] Step 3:

[0163] Preparation of 3-hydroxy-1-(pyridin-2-yl)hexahydropyridin-2-one

[0164] 3-{[dimethyl(2-methylprop-2-yl)silyl]oxy}-1-(pyridin-2-yl)hexahydropyridin-2-one (450 mg, 1.468 mmol) was dissolved in THF (10 mL), and TBAF (2.2 mL, 2.202 mmol, 1 M) was added. The reaction mixture was stirred at 25 °C for 1 hour. The reaction mixture was neutralized with saturated ammonia (10 mL) and extracted three times with ethyl acetate (EA) (20 mL each time). The organic layers were combined, dried over sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (dichloromethane:methanol = 94:6) to give a yellow oil (150 mg, 47% yield). LC / MS: 192.9 [M+H] + .

[0165] Step 4:

[0166] Preparation of N-[(1R,2R)-2-methoxycyclobutyl]-7-(methylamino)-5-{[2-oxoylide-1-(pyridin-2-yl)hexahydropyridin-3-yl]oxy}pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0167] A mixture of 3-hydroxy-l-(pyridin-2-yl)piperidin-2-one (50 mg, 0.122 mmol), {[5-chloro-3-({[(lR,2R)-2-methoxycyclobutyl]amino}carbonyl)pyrazolo[l,5- a]pyrimidin-7-yl](methyl)amino}methane acid-2-methylpropan-2-yl ester (47 mg, 0.243 mmol) and Cs2C03(119 mg, 0.365 mmol) was dissolved in toluene (5 mL) and stirred at 100 °C for 16 h. After the reaction mixture was concentrated under reduced pressure, purification by Genal-Prep-HPLC (acetonitrile: water (0.1% FA) = 10-40) and separation by SFC (Chromatographic column: chiralpak-OJ, mobile phase: CO2-MEOH (DEA)) gave two enantiomers 3-P1 and 3-P2 (P1: 5 mg, P2: 5.8 mg, yield 6.96%) as white solids.

[0168] 3-P1 is 5-{[(3S)-2-oxo-l-(pyridin-2-yl)hexahydropyridin-3-yl]oxy}-N-[(lR,2R)-2- methoxycyclobutyl]-7-(methylamino)pyrazolo[l,5-a]pyrimidine-3-carboxamide or 5-{[(3R)-2-oxo-l-(pyridin-2-yl)hexahydropyridin-3-yl]oxy}-N-[(lR,2R)-2- methoxycyclobutyl]-7-(methylamino)pyrazolo[l,5-a]pyrimidine-3-carboxamide. It is a single configuration white solid.

[0169] LC / MS: 465.8 [M+H] + . 1 HNMR (400 MHz, DMSO) δ 8.57 (d, J = 4.8 Hz, 1H), 8.50 (d, J = 3.8 Hz, 1H), 8.40 (s, 1H), 8.15 (d, J = 9.0 Hz, 1H), 7.85 - 7.77 (m, 2H), 7.27 - 7.20 (m, 1H), 6.62 (s, 1H), 6.53 (s, 1H), 4.20 - 4.14 (m, 2H), 3.96-3.92 (m, 1H), 3.39-3.35 (m, 1H), 3.12 (s, 3H), 3.05 (d, J = 4.8 Hz, 3H), 2.40 (d, J = 8.8 Hz, 1H), 2.18 - 2.04 (m, 4H), 1.79-1.76 (m, 1H), 1.66-1.64 (m, 1H), 1.35 - 1.28 (m, 1H), 0.92 - 0.85 (m, 1H).

[0170] 3-P2 is 5-{[(3R)-2-oxo-1-(pyridin-2-yl)hexahydropyridin-3-yl]oxy}-N-[(1R,2R)-2- methoxycyclobutyl]-7-(methylamino)pyrazolo[1,5-a]pyrimidine-3-carboxamide or 5-{[(3S)-2- oxo-1-(pyridin-2-yl)hexahydropyridin-3-yl]oxy}-N-[(1R,2R)-2-methoxycyclobutyl]-7- (methylamino)pyrazolo[1,5-a]pyrimidine-3-carboxamide. It is a single configuration white solid.

[0171] LC / MS: 465.8 [M+H] + . 1 H NMR (400 MHz, DMSO) δ 8.57 (d, J = 5.1 Hz, 1H), 8.50 (d, J = 4.5 Hz, 1H), 8.40 (s, 1H), 8.15 (d, J = 9.0 Hz, 1H), 7.86 - 7.82 (m, 2H), 7.29 - 7.24 (m, 1H), 6.62 (s, 1H), 6.55 (s, 1H), 4.19 - 4.13 (m, 2H), 3.97 (d, J = 6.2 Hz, 1H), 3.17 (d, J = 7.5 Hz, 1H), 3.05 (d, J = 4.9 Hz, 3H), 3.00 (s, 3H), 2.39 - 2.30 (m, 1H), 2.19 - 2.10 (m, 4H), 1.96 (d, J = 9.4 Hz, 1H), 1.63 (d, J = 9.8 Hz, 1H), 1.31 - 1.29 (m, 1H), 1.11 - 1.05 (m, 1H).

[0172] Synthesis of compound 4

[0173] N-[(1R,2R)-2-methoxycyclobutyl]-7-(methylamino)-5-[(1-phenylhexahydropyridin-3- yl)oxy]pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0174] Step 1: Preparation of ethyl 5-chloro-7-(2,2-dimethyl-4-oxo-5-aza-3-oxahex-5- yl)pyrazolo[1,5-a]pyrimidine-3-carboxylate

[0175] Ethyl 5-chloro-7-(methylamino)pyrazolo[l,5-a]pyrimidine-3-carboxylate (800 mg, 3.141 mmol), (Boc)20 (1371 mg, 6.283 mmol), triethylamine (TEA) (954 mg, 9.424 mmol), DMAP (38 mg, 0.314 mmol) were dissolved in 1,4-dioxane (15 mL) and stirred at 25 °C for 16 h. The reaction mixture was diluted with 50 mL of water and extracted with ethyl acetate (EA) three times (30 mL each time). The organic layer was combined, dried over sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (dichloromethane:methanol = 97:3) to give the product as a white solid (800 mg, yield 68%). LC / MS: 354.7 [M+H] + .

[0176] Step 2:

[0177] Preparation of ethyl 7-(methylamino)-5-[(l-phenylhexahydropyridin-3- yl)oxy]pyrazolo[l,5-a]pyrimidine-3-carboxylate

[0178] Ethyl 7-(2,2-dimethyl-4-oxo-5-aza-3-oxahex-5-yl)-5-[(l- phenylhexahydropyridin-3-yl)oxy]pyrazolo[l,5-a]pyrimidine-3-carboxylate (190 mg, 0.383 mmol) was dissolved in dichloromethane (5 mL), and then trifluoroacetic acid (TFA) (1 mL) was added. The reaction solution was stirred at 25 °C for 16 h. The reaction was completed, and the reaction solution was concentrated under reduced pressure. The residue was purified by column chromatography (dichloromethane:methanol = 97:3) to give the product as a white solid (200 mg, yield 40%). LC / MS: 495.8 [M+H] + .

[0179] Step 3:

[0180] Preparation of ethyl 7-(methylamino)-5-[(l-phenylhexahydropyridin-3- yl)oxy]pyrazolo[l,5-a]pyrimidine-3-carboxylate

[0181] Ethyl 7-(2,2-dimethyl-4-oxo-5-aza-3-oxahex-5-yl)-5-[(l- phenylhexahydropyridin-3-yl)oxy]pyrazolo[l,5-a]pyrimidine-3-carboxylate (190 mg, 0.383 mmol) was dissolved in dichloromethane (5 mL), and then trifluoroacetic acid (TFA)

[0182] (1 mL). The reaction solution was stirred at 25 °C for 1 hour, and the reaction solution was concentrated to get the crude product (200 mg, yellow oil). LC / MS: 395.8 [M+H] + .

[0183] Step 4:

[0184] Preparation of 7-(methylamino)-5-[(1-phenylhexahydropyridin-3- yl)oxy]pyrazolo[1,5-a]pyrimidine-3-carboxylic acid

[0185] Ethyl 7-(methylamino)-5-[(1-phenylhexahydropyridin-3-yl)oxy]pyrazolo[1,5- a]pyrimidine-3-carboxylate (150 mg, 0.266 mmol) was dissolved in dioxane (10 mL), then NaOH aq. (2.5 M, 10 mL) was added, and the reaction solution was stirred at 50 °C for 48 hours. After concentration under reduced pressure, the mixture was diluted with water and adjusted to pH = 6 with hydrochloric acid. Extraction with ethyl acetate (EtOAc) (3 times 20 mL), the organic phase was dried over sodium sulfate, and concentrated under reduced pressure to obtain the crude product (95 mg, yield 88%) as a yellow solid. LC / MS: 368.1 [M+H] + .

[0186] Step 5:

[0187] Preparation of N-[(1R,2R)-2-methoxycyclobutyl]-7-(methylamino)-5-[(1- phenylhexahydropyridin-3-yl)oxy]pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0188] 7-(methylamino)-5-[(1-phenylhexahydropyridin-3-yl)oxy]pyrazolo[1,5- a]pyrimidine-3-carboxylic acid (187 mg, 0.509 mmol), TCFH (306 mg, 1.090 mmol), and NMI (179 mg, 2.180 mmol) were dissolved in dichloromethane (10 mL), then (1R,2R)-2-methoxycyclobutan-1-amine (50 mg, 0.363 mmol) was added, and the mixture was stirred at 25 °C for 2 hours. After the reaction was completed, the reaction solution was concentrated, and the product Cmpd4 (30 mg, yield 18%) was purified by Genal-Prep-HPLC. LC / MS: 450.8 [M+H] + .

[0189] 1H NMR (400 MHz, DMSO) δ 8.26 (s, 1H), 8.19 (d, J = 4.9 Hz, 1H), 8.03 (dd, J = 8.6, 5.5 Hz, 1H), 7.24 - 7.16 (m, 2H), 7.00 - 6.91 (m, 2H), 6.77 (td, J = 7.2, 3.7 Hz, 1H), 5.67 (s, 1H), 5.22 (s, 1H), 4.20 - 4.08 (m, 1H), 3.90 (t, J = 11.4 Hz, 1H), 3.62 - 3.48 (m, 1.7H), 3.29 - 3.21 (m, 0.7H), 3.18 (s, 1.5H), 3.01 (s, 1.5H), 2.99 - 2.85 (m, 4.6H), 2.24 - 2.15 (m, 1H), 2.05 - 1.89 (m, 1.5H), 1.87 - 1.79 (m, 0.5H), 1.79 - 1.63 (m, 3H), 1.39 - 1.25 (m, 1.5H), 1.04 - 0.93 (m, 0.5H).

[0190] Synthesis of compound 5

[0191] N-[(1R,2R)-2-methoxycyclobutyl]-7-(methylamino)-5-[(2-oxo-1-phenyltetrahydro-1H- pyrrol-3-yl)amino]pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0192] Step 1: Into a microwave vial was placed { [5-chloro-3-({ [(1R,2R)-2- methoxycyclobutyl] amino} carbonyl)pyrazolo[1,5-a]pyrimidin-7-yl] (methyl) amino}methane acid-2-methylprop-2-yl ester (123 mg, 0.3 mmol), 3-amino-1-phenyltetrahydropyrrol-2-one (82 mg), palladium acetate (12 mg), Sphos (20 mg), cesium carbonate (200 mg) and toluene (3 mL), which was replaced 3 times under nitrogen protection, and then the mixture was subjected to microwave reaction at 100 °C for 1.5 hours. After cooling to room temperature, LCMS monitoring showed no starting material remained. The toluene was removed by concentration, 20 mL of water and 20 mL of ethyl acetate were added, and after stirring evenly, it was allowed to stand to separate into two layers. The aqueous phase was extracted with ethyl acetate twice (20 mL each time), and the combined organic phase was dried over anhydrous sodium sulfate. After concentration, pre-TLC gave compound 5-1, 103 mg in total, with a yield of 64%. LC / MS: 550.2 [M+H] + .

[0193] Step 2: Into a 50 mL vial was placed compound 5-1 (103 mg) and 15 mL of 20% trifluoroacetic acid in dichloromethane, the reaction was stirred at room temperature for 2 h, spot plate showed no starting material left. Concentrated to remove DCM, added 10 ml of saturated aqueous sodium bicarbonate solution, extracted with ethyl acetate 3 times (20 mL each), combined the organic phase, dried over anhydrous sodium sulfate, concentrated Pre-TLC to give compound 5 (25 mg) as a light yellow solid. Yield 28.8%. LC / MS: 450.2 [M+H] + .

[0194] 1 H NMR (400 MHz, CDCl3) δ 8.29 (s, 1H), 8.21 (t, J = 9.8 Hz, 1H), 7.73 - 7.65 (m, 2H), 7.47 - 7.40 (m, 2H), 7.23 (t, J = 7.4 Hz, 1H), 6.17 (s, 1H), 5.75 (s, 1H), 5.28 (s, 1H), 4.70 (q, J = 11.0, 9.0 Hz, 1H), 4.47 (h, J = 8.0 Hz, 1H), 3.94 (dd, J = 9.7, 4.8 Hz, 2H), 3.79 (q, J = 7.5 Hz, 1H), 3.36 (d, J = 2.8 Hz, 3H), 3.02 (dd, J = 5.3, 1.1 Hz, 3H), 2.28 (q, J = 9.6 Hz, 1H), 1.74 - 1.62 (m, 3H).

[0195] Synthesis of compound 6

[0196] N-[(1R,2R)-2-methoxycyclobutyl]-7-(methylamino)-5-{[2-oxo-1-(pyridin-2-yl)tetrahydro-1H-pyrrol-3-yl]amino}pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0197] Step 1: Into a reaction vial was placed { [5-chloro-3-({ [(1R,2R)-2- methoxycyclobutyl] amino} carbonyl)pyrazolo[1,5-a]pyrimidin-7-yl] (methyl) amino} methanol 2-methylpropan-2-yl ester (200 mg), (3S)-3-amino-1-(pyridin-2-yl) tetrahydropyrrol-2-one (130 mg), Pd2(dba)3(80 mg), Xantphos (110 mg), cesium carbonate (500 mg), and 1,4-dioxane (20 mL) was purged with nitrogen 3 times, and heated at 80 °C for 2 h. The reaction mixture was cooled to room temperature. The 1,4-dioxane was removed by concentration. The reaction mixture was diluted with 100 mL of water and 100 mL of ethyl acetate. The mixture was stirred and allowed to separate into layers. The aqueous layer was extracted with ethyl acetate (3 x 30 mL). The combined organic layers were dried over sodium sulfate and concentrated. The residue was purified by Pre-TLC to give compound 6-1 (80 mg, 31% yield).

[0198] Step 2: Into a 50 mL vial was placed compound 6-1 (80 mg) and 10 mL of 20% trifluoroacetic acid in dichloromethane. The reaction mixture was stirred at room temperature for 2 h. The reaction mixture was concentrated. The residue was diluted with 10 mL of saturated aqueous sodium bicarbonate solution. The mixture was stirred and allowed to separate into layers. The aqueous layer was extracted with ethyl acetate (3 x 30 mL). The combined organic layers were dried over sodium sulfate and concentrated. The residue was purified by Pre-TLC to give compound 6 (7 mg, 12% yield). LC / MS: 451.2 [M+H] + .

[0199] 1 H NMR (400 MHz, CDCl3) δ 8.39 (d, J = 5.0, 2.0 Hz, 1H), 8.30 - 8.20 (m, 2H), 7.86 (ddt, J = 8.4, 4.0, 1.0 Hz, 1H), 7.66 (dd, J = 7.3, 2.0, 1.1 Hz, 1H), 7.11 (ddt, J = 7.3, 4.8, 1.0 Hz, 1H), 6.14 (d, J = 22.9 Hz, 2H), 5.21 (s, 1H), 4.79 (tt, J = 11.6, 5.2 Hz, 1H), 3.81 - 3.72 (m, 2H), 3.37 (d, J = 10.4 Hz, 3H), 3.01 (d, J = 5.2 Hz, 3H), 2.93 (d, J = 12.8, 6.4 Hz, 1H), 2.44 - 2.24 (m, 1H), 2.22 - 2.07 (m, 2H), 1.83 - 1.57 (m, 2H), 1.41 - 1.33 (m, 1H).

[0200] Synthesis of compound 7

[0201] N-[(1R,2R)-2-methoxycyclobutyl]-7-(methylamino)-5-[(1-phenyltetrahydro-1H- pyrrol-3-yl)amino]pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0202] Step 1: Into a microwave vial was placed {[5-chloro-3-({[(1R,2R)-2- methoxycyclobutyl]amino}carbonyl)pyrazolo[1,5-a]pyrimidin-7-yl](methyl)amino}methane acid-2-methylpropan-2-yl ester (123 mg, 0.3 mmol), 1-phenyltetrahydro pyrrol-3-amine (75 mg), palladium acetate (12 mg), Sphos (20 mg), cesium carbonate (200 mg) and toluene (3 mL), and was purged with nitrogen 3 times. The reaction was heated at 100 °C for 1.5 h in the microwave. The reaction was cooled to room temperature. LCMS showed no starting material left. The toluene was removed by concentration. 20 mL water and 20 mL ethyl acetate were added. The mixture was stirred well and was allowed to separate. The aqueous phase was extracted with ethyl acetate 2 times (20 mL each time). The organic phases were combined and dried over sodium sulfate. The solvent was removed by concentration. The residue was purified by column chromatography to give compound 7-1 (86 mg, 53.3% yield). LC / MS: 536.1 [M+H] + .

[0203] Step 2: Into a 50 mL vial was placed compound 7-1 (86 mg) and 15 mL of 20% trifluoroacetic acid in dichloromethane. The reaction was stirred at room temperature for 2 h. The reaction was spotted on a TLC plate and showed no starting material left. The solvent was removed by concentration. 10 mL of saturated aqueous sodium bicarbonate solution was added. The mixture was extracted with ethyl acetate 3 times (20 mL each time). The organic phases were combined and dried over sodium sulfate. The solvent was removed by concentration. Pre-TLC gave compound 7 (20 mg) as a light yellow solid. Yield 27.8%. LC / MS: 436.2 [M+H] + .

[0204] 1H NMR (400 MHz, CDC13) δ 8.29 (s, 1H), 8.21 (t, J = 9.8 Hz, 1H), 7.73 - 7.65 (m, 2H), 7.47 - 7.40 (m, 2H), 7.23 (t, J = 7.4 Hz, 1H), 6.17 (s, 1H), 5.75 (s, 1H), 5.28 (s, 1H), 4.70 (q, J = 11.0, 9.0 Hz, 1H), 4.47 (s, 1H), 4.21 (h, J = 8.0 Hz, 2H), 3.94 (dd, J = 9.7, 4.8 Hz, 2H), 3.79 (q, J = 7.5 Hz, 1H), 3.36 (d, J = 2.8 Hz, 3H), 3.02 (dd, J = 5.3, 1.1 Hz, 3H), 2.28 (q, J = 9.6 Hz, 1H), 1.74 - 1.62 (m, 3H).

[0205] Synthesis of compound 11

[0206] Step 1: Into a reaction vial was placed compound {[5-chloro-3-({[(1R,2R)-2- methoxycyclobutyl]amino}carbonyl)pyrazolo[1,5-a]pyrimidin-7-yl](methyl)amino}methane acid-2-methylpropan-2-yl ester (120 mg), compound 1-phenylhexahydropyridin-3-amine (130 mg), Pd2(dba)3(80 mg), Xantphos (110 mg), cesium carbonate (500 mg) and 1,4-dioxane (20 mL), which was replaced 3 times under nitrogen protection, and the reaction was carried out at 80 °C for 2 hours. After cooling to room temperature, 1,4-dioxane was removed by concentration, 100 mL of water and 100 mL of ethyl acetate were added, and after stirring evenly, it was allowed to stand and separate, the aqueous phase was extracted with ethyl acetate for 3 times (30 mL each time), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated by scraping to obtain compound 11-1, 50 mg in total, with a yield of 33%.

[0207] Step 2: Into a 50 mL vial was placed compound 11-1 (50 mg) and 6 mL of 20% trifluoroacetic acid in dichloromethane, and the reaction was carried out at room temperature for 2 hours. Spotting showed that no starting material remained. After removing DCM by concentration, 10 mL of saturated aqueous sodium bicarbonate solution was added, and the mixture was extracted with ethyl acetate for 3 times (30 mL each time), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated by scraping to obtain compound 11 (5 mg), with a yield of 11%. LC / MS: 450.3 [M+H]+.

[0208] 1H NMR (400 MHz, CDC13) δ 8.29 (s, 1H), 8.21 (t, J = 9.8 Hz, 1H), 7.73 - 7.65 (m, 2H), 7.47 - 7.40 (m, 2H), 7.23 (t, J = 7.4 Hz, 1H), 6.17 (s, 1H), 5.75 (s, 1H), 5.28 (s, 1H), 4.70 (q, J = 11.0, 9.0 Hz, 1H), 4.47 (s, 1H), 4.21 (h, J = 8.0 Hz, 2H), 3.94 (dd, J = 9.7, 4.8 Hz, 2H), 3.75 (q, J = 7.5 Hz, 1H), 3.36 (d, J = 2.8 Hz, 3H), 3.02 (dd, J = 5.3, 1.1 Hz, 3H), 2.26 (q, J = 9.6 Hz, 2H), 1.72 - 1.62 (m, 4H).

[0209] Synthesis of compound 13

[0210] Following the procedure for the synthesis of Reference Compound 3, the alcohol used was (3S)-3-hydroxy-1-(pyridin-2-yl)tetrahydropyridin-2-one to give Compound 13, 12 mg, 34% yield. LC / MS: 452.2 [M+H] + .

[0211] 1 H NMR (400 MHz, DMSO) δ 8.53 (d, J = 5.1 Hz, 1H), 8.50 (d, J = 4.5 Hz, 1H), 8.40 (s, 1H), 8.12 (d, J = 9.0 Hz, 1H), 7.83 - 7.80 (m, 2H), 7.29 - 7.24 (m, 1H), 6.62 (s, 1H), 6.51 (s, 1H), 4.19 - 4.13 (m, 2H), 3.92 (d, J = 6.2 Hz, 1H), 3.17 (d, J = 7.5 Hz, 1H), 3.05 (d, J = 4.9 Hz, 3H), 3.00 (s, 3H), 2.39 - 2.30 (m, 1H), 2.17 - 2.10 (m, 2H), 1.96 (d, J = 9.4 Hz, 1H), 1.61 (d, J = 9.8 Hz, 1H), 1.35 - 1.29 (m, 1H), 1.11 - 1.05 (m, 1H).

[0212] Synthesis of compound 17

[0213] Following the procedure for the synthesis of Reference Compound 5, the amine used was (3S)-3-amino-1-(pyridin-3-yl)tetrahydropyrrol-2-one to give Compound 17, 16 mg, 37% yield. LC / MS: 451.2 [M+H]+ .

[0214] 1 H NMR (400 MHz, CDC13) δ 8.33 (s, 1H), 8.26 (t, J = 9.8 Hz, 1H), 7.73 - 7.65 (m, 2H), 7.47 - 7.40 (m, 2H), 7.27 (t, J = 7.4 Hz, 1H), 6.11 (s, 1H), 5.75 (s, 1H), 5.28 (s, 1H), 4.70 (q, J = 11.0, 9.0 Hz, 1H), 4.47 (h, J = 8.0 Hz, 1H), 3.94 (dd, J = 9.7, 4.8 Hz, 2H), 3.79 (q, J = 7.5 Hz, 2H), 3.33 (d, J = 2.8 Hz, 3H), 3.01 (dd, J = 5.3, 1.1 Hz, 3H), 2.25 (q, J = 9.6 Hz, 2H), 1.71 - 1.62 (m, 3H).

[0215] Synthesis of compound 18

[0216] Following the procedure for the synthesis of compound 5, using 3-amino-1- (pyridin-3-yl)hexahydropyridin-2-one as the amine, compound 18 was obtained, 12 mg, 31% yield. LC / MS: 465.3 [M+H] + .

[0217] 1 H NMR (400 MHz, CDC13) δ 8.33 (s, 1H), 8.26 (t, J = 9.8 Hz, 1H), 7.73 - 7.65 (m, 2H), 7.47 - 7.40 (m, 2H), 7.27 (t, J = 7.4 Hz, 1H), 6.11 (s, 1H), 5.75 (s, 1H), 5.28 (s, 1H), 4.70 (q, J = 11.0, 9.0 Hz, 1H), 4.47 (h, J = 8.0 Hz, 1H), 3.94 (dd, J = 9.7, 4.8 Hz, 2H), 3.79 (q, J = 7.5 Hz, 2H), 3.33 (d, J = 2.8 Hz, 3H), 3.01 (dd, J = 5.3, 1.1 Hz, 3H), 2.25 (q, J = 9.6 Hz, 2H), 1.71 - 1.62 (m, 3H).

[0218] Synthesis of compound 19

[0219] Following the procedure for the synthesis of compound 5, using 3-amino-1- (pyridin-3-yl)hexahydropyridin-2-one as the amine, compound 18 was obtained, 12 mg, 31% yield. LC / MS: 465.3 [M+H] +

[0220] 1 H NMR (400 MHz, CDC13) δ 8.27 (s, 1H), 8.19 (t, J = 9.8 Hz, 1H), 7.73 - 7.65 (m, 2H), 7.49 - 7.40 (m, 1H), 7.21 (t, J = 7.4 Hz, 1H), 6.17 (s, 1H), 5.79 (s, 1H), 5.28 (s, 1H), 4.70 (q, J = 11.0, 9.0 Hz, 1H), 4.47 (h, J = 8.0 Hz, 1H), 3.94 (dd, J = 9.7, 4.8 Hz, 2H), 3.77 (q, J = 7.5 Hz, 2H), 3.36 (s, 3H), 3.02 (dd, J = 5.3, 1.1 Hz, 3H), 2.28 (q, J = 9.6 Hz, 2H), 1.74 - 1.62 (m, 3H).

[0221] Synthesis of compound 20

[0222] Following the procedure for the synthesis of Reference Compound 2, using 3-amino-1- (1,3-thiazinan-5-yl)hexahydropyridin-2-one as the amine, compound 20 was obtained, 6 mg, 25% yield. LC / MS: 471.2 [M+H] + .

[0223] 1 H NMR (400 MHz, CDC13) δ 8.27 (s, 1H), 8.19 (t, J = 9.8 Hz, 1H), 7.73 - 7.65 (m, 2H), 7.49 - 7.40 (m, 1H), 7.21 (t, J = 7.4 Hz, 1H), 6.17 (s, 1H), 5.79 (s, 1H), 5.28 (s, 1H), 4.70 (q, J = 11.0, 9.0 Hz, 1H), 4.47 (h, J = 8.0 Hz, 1H), 3.94 (dd, J = 9.7, 4.8 Hz, 2H), 3.77 (q, J = 7.5 Hz, 2H), 3.36 (s, 3H), 3.02 (dd, J = 5.3, 1.1 Hz, 3H), 2.28 (q, J = 9.6 Hz, 2H), 1.74 - 1.62 (m, 3H).

[0224] Synthesis of compound 21

[0225] Following the procedure for the synthesis of Reference Compound 2, using 3-amino-1- (1,3-thiazinan-5-yl)hexahydropyridin-2-one as the amine, compound 20 was obtained, 6 mg, 25% yield. LC / MS: 471.2 [M+H] + .

[0226] ​1 H NMR (400 MHz, CDC13) δ 8.31 (t, J = 9.8 Hz, 1H), 7.78 - 7.65 (m, 1H), 7.47 - 7.41 (m, 1H), 7.23 (t, J = 7.4 Hz, 1H), 6.17 (s, 1H), 5.75 (s, 1H), 5.21 (s, 1H), 4.70 (q, J = 11.0, 9.0 Hz, 1H), 4.47 (h, J = 8.0 Hz, 1H), 3.94 (dd, J = 9.7, 4.8 Hz, 2H), 3.79 (s, 3H), 3.32 (s, 3H), 3.01 (dd, J = 5.3, 1.1 Hz, 3H), 2.23 - 2.15 (q, J = 9.6 Hz, 4H), 1.77 - 1.15 (q, J = 9.6 Hz, 2H).

[0227] Other compounds were prepared by similar procedures as described previously:

[0228] The beneficial effects of the present application are demonstrated by the following experiments.

[0229] I. Experimental Methods

[0230] 1. TYK2 / JAK1 JH2 in vitro enzyme binding experiment

[0231] Experimental instruments: Envision microplate reader (PerkinElmer), Echo (LABCYTE)

[0232] Test method: In this experiment, the method of fluorescence resonance energy transfer (TR-FRET) was used to test the inhibitory effect of the compound on TYK2 / JAK1 JH2 pseudokinase.

[0233] Experimental procedure: Dissolve compound to 10 mM stock concentration in DMSO, dilute compound in DMSO to different concentration gradient, use Echo instrument to transfer 75 nL of 200x final concentration compound dilution to 384 experimental plate. Prepare 1x working solution of the following table: HEPES pH 7.5 final concentration 20 mM, MgCl2 final concentration 10 mM, Brij-35 final concentration 0.015%, DTT final concentration 2 mM, BSA final concentration 50 ug / mL. Use 1x working solution to configure 3x final concentration of TYK2 JH2 and JAK1 JH2 pseudo kinase, Tb antibody, Tracer respectively. Add 5 uL of TYK2 JH2 or JAK1 JH2 pseudo kinase to 384 well experimental plate for positive control well and compound well, replace with same volume of 1X experimental working solution for negative control well, centrifuge at 1000 rpm for 30 seconds. Add 5 uL of Tb antibody to 384 well experimental plate, centrifuge at 1000 rpm for 30 seconds. Add 5 uL of Tracer to 384 well experimental plate, centrifuge at 1000 rpm for 30 seconds. Incubate at room temperature for 60 minutes, then incubate overnight at 4 degrees. Finally read 495 nm / 520 nm fluorescence signal ratio on Envision plate reader (PerkinElmer).

[0234] Experimental data processing method: Use XLfit software written by IDBS company, integrated in Microsoft Excel environment, to process and analyze test data. First, calculate the average value of positive control well and negative control well reaction signal respectively, then calculate the reaction inhibition rate of each compound well according to the formula "single well inhibition rate % = 100-(single well signal value-negative control group signal average value) / (positive control group signal average value-negative control group signal average value)". Then import the concentration and corresponding inhibition rate data into XLfit software, use Dose Response One Site 205 model in the software, use four parameter method inhibition rate-concentration curve to fit, and calculate the IC 50 value of the compound.

[0235] 2, Test the inhibition of JAK1 JH1 / JAK2 JH1 / JAK3 JH1 / TYK2 JH1 kinase by test compound

[0236] Experimental instrument: Envision plate reader (PerkinElmer), Echo (LABCYTE)

[0237] Test method: This experiment uses the method of fluorescence resonance energy transfer (TR-FRET) to test the inhibition of JAK1 / JAK2 / JAK3 / TYK2 JH1 kinase activity by compound.

[0238] Experimental procedure: Dissolve compounds in DMSO to 10 mM stock concentration, dilute compounds in DMSO to different concentration gradient, transfer 100 nL of 100x final concentration compound dilution to 384 experimental plate using Echo instrument. Prepare 1x working solution as follows. JAK1 JH1 reaction buffer: 5X Enzymatic buffer final concentration 1X, MgCl2 final concentration 5 mM, EGTA final concentration 0.625 mM, SEB final concentration 0.06 mM, Brij-35 final concentration 0.01%, DTT final concentration 1 mM. JAK2 / 3 JH1 reaction buffer: 5X Enzymatic buffer final concentration 1X, MgCl2 final concentration 5 mM, DTT final concentration 1 mM. TYK2 JH1 reaction buffer: 5X Enzymatic buffer final concentration 1X, MgCl2 final concentration 5 mM, MnCl2 final concentration 1 mM, SEB final concentration 0.0125 mM, DTT final concentration 1 mM. Compound and kinase pre-incubation: prepare 2x JAK1 / JAK2 / JAK3 / TYK2 JH1 enzyme solution using corresponding reaction buffer, transfer compounds to 384 experimental plate using automated micropipetting system at 100 nL per well (negative and positive controls replaced by DMSO), add 5 pL of 2x JAK1 / JAK2 / JAK3 / TYK2 JH1 enzyme solution to each well (compound wells and positive control wells) or reaction buffer (negative control wells), centrifuge to mix, incubate at 25 °C room temperature for 15 minutes. Enzyme reaction: prepare 2x TK-Sub-biotin substrate and ATP mixture solution of JAK1 / JAK2 / JAK3 / TYK2 JH1 enzyme using corresponding reaction buffer, add 5 pL of 2x TK-Sub-biotin substrate and ATP mixture solution to each well, centrifuge to mix, react at 25 °C for 45 minutes (JAK1 / JAK2 JH1) or 60 minutes (JAK3 / TYK2 JH1). Detection: prepare detection mixture of TK Antibody-Eu and streptavidin-XL665 using detection buffer, add 10 pL of detection mixture to each well of the above 384 experimental plate, centrifuge to mix, react at 25 °C for 60 minutes (JAK1 / JAK2 JH1) or 120 minutes (JAK3 / TYK2 JH1). After reaction, transfer to 4 °C refrigerator for overnight incubation, read fluorescence value on Envision 2104 Multilabel Reader (340 nm excitation, detect 665 nm and 615 nm emission, 665 nm / 615 nm fluorescence ratio is the raw data of well reaction signal value).

[0239] Data processing and analysis: The data processing and analysis of the kinase activity detection experiment was performed by using XLfit software written by IDBS company and integrated in Microsoft Excel environment. First, the average values of the positive control and negative control wells were calculated, respectively. Then, the inhibition rate of each compound well could be calculated according to the formula "inhibition rate of single well = (1-(signal value of single well-negative control signal average value) / (positive control signal average value-negative control signal average value))x100%". Finally, the concentration and corresponding inhibition rate data were imported into XLfit software, and the IC50value of the compound was calculated by using the Dose Response One Site 205 model in the software and the four-parameter inhibition rate-concentration curve fitting. 50

[0240] 3. FACS detection of the inhibitory effect of the compound on the expression of pSTAT5 in CD3+ cells

[0241] Experimental instrument: Beckman CytoFlex S flow cytometer.

[0242] Experimental method: In this experiment, the flow cytometry method was used to detect the inhibitory effect of the compound on the expression of phosphorylated STAT5 in CD3+ cells in human PBMC.

[0243] ​Experimental procedure: Compound was prepared as 10 mM solution in DMSO, diluted in DMSO to 500x final concentration in different concentration gradient of DMSO solution, 5 uL of diluted compound in DMSO was transferred to 120 uL of DPBS containing 0.1% BSA solution, positive and negative control groups were set up, final concentration of 0.2% DMSO in positive and negative control groups. 0.5 million human PBMC cells were added to each well of a 96-well cell culture plate with a volume of 66.5 uL, 3.5 uL of diluted compound was added, mixed, and incubated in a 37°C incubator for 60 minutes. IFN-alpha was diluted to 600 ng / mL in DPBS containing 0.1% BSA, and PE-anti-hCD3 antibody was diluted 4-fold. After the above 60-minute incubation was complete, 6 uL of PE-anti-hCD3 antibody was added to each well, 4 uL of diluted IFN-alpha was added to each well of the positive control wells and the compound wells (0.1% BSA in DPBS was used instead of the negative control group), and incubated in a 37°C incubator for 30 minutes. All cells were transferred to a 96-well deep well plate, 1 mL of 37°C preheated Lyse / fix buffer was added, and incubated at 37°C for 10 minutes in the dark. After centrifugation at 600g for 5 minutes, the supernatant was discarded, washed twice with 1 mL of DPBS and centrifuged, 0.4 mL of Permbuffer III was added to the cell pellet, and incubated at 4°C for 30 minutes in the dark. After centrifugation at 600g for 5 minutes, the supernatant was discarded, washed twice with 1 mL of Staining buffer (0.2% BSA + 1 mM EDTA + PBS) and centrifuged, APC anti-human pSTAT5 antibody was diluted 200-fold in Staining buffer, 100 uL per well was added to the cell well, mixed, and incubated at room temperature for 40 minutes. Washed twice with Staining buffer, 1 mL per well, centrifuged at 600g for 5 minutes, and resuspended the cell pellet in 300 uL of Staining buffer after discarding the supernatant. Sample analysis was performed on a Beckman CytoFlex flow cytometer.

[0244] Experimental data processing method: Flowjo software was used to process and analyze the data of phospho-STAT5 detection experiment. First, the CD3 cells with high signal expression were circled on the software, and then the mean fluorescence intensity of phospho-STAT5 expression in CD3 cells was analyzed. According to the mean fluorescence signal values of positive control group and negative control group corresponding to each plate, the inhibition rate of phospho-STAT5 of each compound well was calculated by the following formula: "single well inhibition rate % = 100- (single well signal value-negative control group signal average value) / (positive control group signal average value-negative control group signal average value) x 100%" to calculate the inhibition rate of each compound well. Then the concentration and corresponding inhibition rate data were imported into GraphPad Prism 5 software, and the Dose Response-inhibition model in the software was used to fit the four parameter inhibition rate-concentration curve, and the IC50 value of the compound was calculated.

[0245] 4. Kinetic solubility experiment

[0246] Experimental instruments: LC / MS / MS, pH meter

[0247] Experimental method: In this experiment, a certain concentration of compound was mixed with the matrix, shaken for 4 hours at 25 degrees, the reaction solution was filtered and diluted, precipitated with the termination solution, and then subjected to LC / MS / MS quantitative analysis. According to the calibration curve, the concentration of the compound dissolved in the matrix can be quantified.

[0248] Experimental operation:

[0249] (1) Prepare buffer solutions of various matrices.

[0250] (2) Dissolve the compound powder in DMSO to prepare a stock solution, and the conventional maximum concentration is 50 mM. If it cannot be completely dissolved, reduce the concentration to completely dissolve.

[0251] (3) Dissolve 2 uL of the stock solution in 198 uL of the matrix, control the DMSO concentration at 1%, and the target concentration of the compound is 100-500 uM. The target concentration of the compound in the matrix is determined by the solubility of the compound in DMSO.

[0252] (4) Shake the above solution in Eppendorf ThermoMixer for 4 hours, and control the temperature at 25°C.

[0253] (5) After 4 hours, filter the above solution, and dilute the filtrate 10 times with DMSO for standby.

[0254] (6) Prepare a set of calibration curves with concentrations of 0.02, 0.06, 0.2, 0.6, 2, 6, 20, and 60 uM using DMSO.

[0255] (7) Take 5 uL of the calibration and diluted filtrate, add to 250 uL of stop solution (acetonitrile solution containing internal standard) and 250 uL of ultrapure water. Vortex well for 1 minute.

[0256] (8) Perform LS / MS / MS analysis on the sample

[0257] Experimental data processing method:

[0258] Quantify the sample diluted 10 times by the calibration curve, and calculate the concentration of the compound dissolved in the matrix.

[0259] II. Experimental results

[0260] Table 1. JAK1 & TYK2 JH2 binding experiment

[0261] Note: A represents IC 50 <50 nM; B represents IC 50 ≥ 50 and < 500 nM; C represents IC 50 ≥ 500 and ≤ 5000 nM; D represents IC 50 > 5000 nM; n / a represents no activity detected at 5000 nM; n.d. represents not detected.

[0262] TAK-279 is a known TYK2 inhibitor capable of binding to TYK2 JH2 (i.e., Compound I-816 disclosed in Chinese Patent Application Publication No. CN111194317A), and has the following structure:

[0263] BMS986165 is a known TYK2 inhibitor capable of binding to TYK2 JH2, and has the following structure:

[0264] The results show that the compounds of the present application exhibit good TYK2 JH2 binding activity, and have no obvious binding to JAK1 JH2. It is shown that the compounds of the present application have excellent TYK2 JH2 selectivity.

[0265] Table 2. Compound solubility

[0266] The results show that the compounds of the present application have excellent solubility, which is significantly better than the control compound TAK-279.

[0267] In conclusion, the present application provides a compound with TYK2 inhibiting effect shown in formula I, a preparation method and use thereof, the compound exhibits good binding activity to TYK2 JH2, can effectively inhibit the TYK2 signal pathway stimulated and activated by IFN-alpha, IL-12 and IL-23 in human PBMC cells, the compound has excellent physical and chemical properties, good permeability and excellent solubility, has significant advantages in preparation development and production and clinical application, and has a wide application prospect in preparation of diseases related to TYK2 activity.

Claims

1. A compound, pharmaceutically acceptable salt thereof, stereoisomer thereof, deuterated form thereof, or solvate thereof, characterized in that, The structure of the compound is shown in formula I: wherein is a 5-membered heteroaromatic ring containing two nitrogen atoms, E1 is selected from CH or N, E2 is selected from CH or N, E3 is selected from CH or N, E4 is selected from CH or N; X is selected from O, S, NH, C 1-6 alkylene, C 2-6 alkenylene, C 2-6 alkynylene, 3-8 membered saturated cycloalkyl, 3-8 membered saturated heterocyclyl; Y is selected from CONH, NHCO, CO, COO, OCO, NH; selected from =0, =S, =CH2, -H, -R 1m , -NR 1c R 1d , -CONR 1c R 1d , -COOR 1e ; R 1m selected from C 1-8 alkyl, C 1-8 alkoxy, halogen; R 1c selected from hydrogen, C 1-8 alkyl, R 1d selected from hydrogen, C 1-8 alkyl, COR 1f , or R 1g is selected from the group consisting of 4-8 membered heteroaromatic ring, 4-8 membered aromatic ring, 3-8 membered saturated cycloalkyl, 3-8 membered saturated heterocyclyl; or, R 1c , R 1d are joined to form a 3-8 membered saturated heterocyclyl; R 1e selected from C 1-8 alkyl, R 1f selected from C 1-8 alkyl, R 1g selected from C 1-8 alkyl; m is selected from 0, 1, 2 or 3; n is selected from 0, 1, 2, 3 or 4; R2are each independently selected from the group consisting of hydrogen, C 1-8 alkyl, C 1-8 alkoxy, halogen; Z is N; A ring is absent or selected from a 4-8 membered heteroaromatic ring, a 4-8 membered aromatic ring, a 3-8 membered saturated cycloalkyl, a 3-8 membered saturated heterocyclyl; a is selected from 0, 1, 2, 3 or 4; R3are each independently selected from the group consisting of hydrogen, halogen, COOR 3a , OCOR 3a , COR 3a , OR 3a , unsubstituted or substituted by one or more than one R 3b ; C 1-8 alkyl, C 2-8 alkenyl, C 2-8 alkynyl, 3-8 membered saturated cycloalkyl, 3-8 membered saturated heterocyclyl; R 3a is selected from the group consisting of C 1-8 alkyl, R 3b is each independently selected from the group consisting of halogen, C 1-8 alkyl; B ring is selected from a 3-8 membered saturated cycloalkyl, a 3-8 membered saturated heterocyclyl; b is selected from 0, 1, 2, 3 or 4; R4are each independently selected from the group consisting of hydrogen, halogen, COOR 4a , OCOR 4a , COR 4a , OR 4a , and the following groups which are unsubstituted or substituted by one or more than one R 4b ; C 1-8 alkyl, C 2-8 alkenyl, C 2-8 alkynyl, 3-8 membered saturated cycloalkyl, 3-8 membered saturated nitrogen containing heterocyclyl; R 4a is selected from the group consisting of C 1-8 alkyl, R 4b is each independently selected from the group consisting of halogen, C 1-8 alkyl; R5is selected from C 1-8 alkyl.

2. The compound, pharmaceutically acceptable salt thereof, stereoisomer thereof, deuterated form thereof or solvate thereof according to claim 1, characterized in that, The structure of the compound is shown in formula II: wherein is a 5-membered heteroaromatic ring containing two nitrogen atoms, E1 is selected from CH or N, E2 is selected from CH or N, E3 is selected from CH or N, E4 is selected from CH or N; X is selected from O, S, NH, C 1-6 alkylene, C 2-6 alkenylene, C 2-6 alkynylene, 3-8 membered saturated cycloalkyl, 3-8 membered saturated heterocyclyl; Y is selected from CONH, NHCO, CO, COO, OCO, NH; selected from =0, =S, =CH2, -H, -R 1m , -NR 1c R 1d , -CONR 1c R 1d , -COOR 1e ; R 1m selected from C 1-8 alkyl, C 1-8 alkoxy, halogen; R 1c selected from hydrogen, C 1-8 alkyl, R 1d selected from hydrogen, C 1-8 alkyl, COR 1f , or R 1g is selected from the group consisting of a 5-6 membered heteroaromatic ring, a phenyl ring, a 5-6 membered saturated cycloalkyl group, a 5-6 membered saturated heterocyclyl group; or, R 1c , R 1d are joined to form a 5-6 membered saturated heterocyclyl group; R 1e selected from C 1-8 alkyl, R 1f selected from C 1-8 alkyl, R 1g selected from C 1-8 alkyl; m is selected from 0, 1, 2 or 3; n is selected from 0, 1, 2, 3 or 4; R2are each independently selected from the group consisting of hydrogen, C 1-8 alkyl, C 1-8 alkoxy, halogen; Z is N; A ring is absent or selected from a 4-8 membered heteroaromatic ring, a 4-8 membered aromatic ring, a 3-8 membered saturated cycloalkyl, a 3-8 membered saturated heterocyclyl; a is selected from 0, 1, 2, 3 or 4; R3are each independently selected from the group consisting of hydrogen, halogen, COOR 3a , OCOR 3a , COR 3a , OR 3a , and the following groups which are unsubstituted or substituted by one or more R 3b ; C 1-8 alkyl, C 2-8 alkenyl, C 2-8 alkynyl, 3-8 membered saturated cycloalkyl, 3-8 membered saturated heterocyclyl; R 3a is selected from the group consisting of C 1-8 alkyl, R 3b is each independently selected from the group consisting of halogen, C 1-8 alkyl; B ring is selected from a 3-8 membered saturated cycloalkyl, a 3-8 membered saturated heterocyclyl; b is selected from 0, 1, 2, 3 or 4; R4are each independently selected from the group consisting of hydrogen, halogen, COOR 4a , OCOR 4a , COR 4a , OR 4a , and the following groups which are unsubstituted or substituted by one or more than one R 4b C 1-8 alkyl, C 2-8 alkenyl, C 2-8 alkynyl, 3-8 membered saturated cycloalkyl, 3-8 membered saturated nitrogen containing heterocyclyl; R 4a is selected from the group consisting of C 1-8 alkyl, R 4b are each independently selected from the group consisting of halogen, C 1-8 alkyl; R5is selected from C 1-8 alkyl.

3. The compound, pharmaceutically acceptable salt thereof, stereoisomer thereof, deuterated form thereof or solvate thereof according to claim 2, characterized in that, The structure of the compound is shown as formula III-1, formula III-1, or formula III-3:

4. The compound, pharmaceutically acceptable salt thereof, stereoisomer thereof, deuterated form thereof or solvate thereof according to claim 3, characterized in that, The selected from =0, =S, =CH2, -H, -R 1m , -NR 1c R 1d , -CONR 1c R 1d , -COOR 1e ; R 1m selected from C 1-6 alkyl; R 1c selected from hydrogen, C 1-6 alkyl, R 1d selected from hydrogen, C 1-6 alkyl, COR 1f , or the following groups, which are unsubstituted or substituted by one or more R 1g 5-6 membered heteroaromatic ring, benzene ring, 5-6 membered saturated cycloalkyl, 5-6 membered saturated heterocyclyl; or, R 1c , R 1d form a 5-6 membered saturated heterocyclyl ring; R 1e selected from C 1-6 alkyl, R 1f selected from C 1-6 alkyl, R 1g selected from C 1-6 alkyl; X is selected from O, S, NH, C 1-6 alkylene, C 2-6 alkenylene, C 2-6 alkynylene, 3-6 membered saturated cycloalkyl, 3-6 membered saturated heterocyclyl; Y is selected from CONH, NHCO, CO, COO, OCO, NH.

5. The compound, pharmaceutically acceptable salt thereof, stereoisomer thereof, deuterated form thereof or solvate thereof according to claim 4, characterized in that, The structure of the compound is shown as Formula IV-1, Formula IV-2, or Formula IV-3: wherein R 1m selected from C 1-6 alkyl; X is selected from O, S, NH, CH2, HC=CH, C≡C, a 3-6 membered saturated cycloalkyl; m is selected from 0, 1 or 2; R2is selected from hydrogen, C 1-6 alkyl; A ring is selected from a 5-6 membered heteroaromatic ring, a benzene ring, a 3-6 membered saturated cycloalkyl, a 3-6 membered saturated heterocyclyl; a is selected from 0, 1, 2, 3 or 4; R3are each independently selected from the group consisting of hydrogen, halogen, COOR 3a , OCOR 3a , COR 3a , OR 3a , and the following groups which are unsubstituted or substituted by one or more R 3b ; C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, 3- to 6-membered saturated cycloalkyl, 3- to 6-membered saturated heterocyclyl; R 3a is selected from the group consisting of C 1-6 alkyl, R 3b is each independently selected from the group consisting of halogen, C 1-6 alkyl; B ring is selected from a 3-6 membered saturated cycloalkyl, a 3-6 membered saturated heterocyclyl; b is selected from 0, 1, 2, 3 or 4; R4are each independently selected from the group consisting of hydrogen, halogen, COOR 4a , OCOR 4a , COR 4a , OR 4a , and the following group, which is unsubstituted or substituted by one or more than one R 4b ; C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, 3-6 membered saturated cycloalkyl, 3-6 membered saturated nitrogen containing heterocyclyl; R 4a is selected from the group consisting of C 1-6 alkyl, R 4b is each independently selected from the group consisting of halogen, C 1-6 alkyl; R5is selected from C 1-6 alkyl.

6. The compound, pharmaceutically acceptable salt thereof, stereoisomer thereof, deuterated form thereof or solvate thereof according to claim 4, characterized in that, The structure of the compound is shown as formula V-1, formula V-2, or formula V-3: wherein R 1m selected from C 1-6 alkyl; X is selected from O, S, NH, CH2, HC=CH, C≡C, a 3-6 membered saturated cycloalkyl; m is selected from 0, 1 or 2; R2is selected from hydrogen, C 1-6 alkyl; A ring is selected from a 5-6 membered heteroaromatic ring, a benzene ring, a 3-6 membered saturated cycloalkyl, a 3-6 membered saturated heterocyclyl; a is selected from 0, 1, 2, 3 or 4; R3 is independently selected from hydrogen, halogen, and COOR. 3a OCOR 3a COR 3a OR 3a Not replaced or replaced by one or more R 3b The following groups are substituted: C 1-6 Alkyl, C 2-6 alkenyl, C 2-6 Alkyne group, 3-6 membered saturated cycloalkyl group, 3-6 membered saturated heterocyclic group; R 3a Selected from C 1-6 Alkyl, R 3b Each is independently selected from halogens, C 1-6 alkyl; B ring is selected from a 3-6 membered saturated cycloalkyl, a 3-6 membered saturated heterocyclyl; b is selected from 0, 1, 2, 3 or 4; R4are each independently selected from the group consisting of hydrogen, halogen, COOR 4a , OCOR 4a , COR 4a , OR 4a , and the following group, which is unsubstituted or substituted by one or more than one R 4b C 1-6 1-6 alkyl, C 2-6 2-6 alkenyl, C 2-6 2-6 alkynyl, 3-6 membered saturated cycloalkyl, 3-6 membered saturated nitrogen containing heterocyclyl; R 4a is selected from the group consisting of C 1-6 1-6 alkyl, R 4b are each independently selected from the group consisting of halogen, C 1-6 1-6 alkyl; R5is selected from C 1-6 alkyl.

7. The compound, pharmaceutically acceptable salt thereof, stereoisomer thereof, deuterated form thereof or solvate thereof according to claim 4, characterized in that, The structure of the compound is shown as formula VI-1, formula VI-2, or formula VI-3: wherein R 1m selected from C 1-6 alkyl; X is selected from O, S, NH, CH2, HC=CH, C≡C, a 3-6 membered saturated cycloalkyl; m is selected from 0, 1 or 2; R2is selected from hydrogen, C 1-6 alkyl; A ring is selected from a 5-6 membered heteroaromatic ring, a benzene ring, a 3-6 membered saturated cycloalkyl, a 3-6 membered saturated heterocyclyl; a is selected from 0, 1, 2, 3 or 4; R3are each independently selected from the group consisting of hydrogen, halogen, COOR 3a , OCOR 3a , COR 3a , OR 3a , and the following groups which are unsubstituted or substituted by one or more R 3b ; C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, 3- to 6-membered saturated cycloalkyl, 3- to 6-membered saturated heterocyclyl; R 3a is selected from the group consisting of C 1-6 alkyl, R 3b is each independently selected from the group consisting of halogen, C 1-6 alkyl; B ring is selected from a 3-6 membered saturated cycloalkyl, a 3-6 membered saturated heterocyclyl; b is selected from 0, 1, 2, 3 or 4; R4are each independently selected from the group consisting of hydrogen, halogen, COOR 4a , OCOR 4a , COR 4a , OR 4a , and the following groups which are unsubstituted or substituted by one or more than one R 4b C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, 3-6 membered saturated cycloalkyl, 3-6 membered saturated nitrogen containing heterocyclyl; R 4a is selected from the group consisting of C 1-6 alkyl, R 4b are each independently selected from the group consisting of halogen, C 1-6 alkyl; R5is selected from C 1-6 alkyl.

8. The compound, pharmaceutically acceptable salt thereof, stereoisomer thereof, deuterated form thereof, or solvate thereof of any one of claims 5-7, wherein, The R 1m Selected from C 1-3 alkyl; X is selected from O, S, NH, CH2, HC=CH, C≡C, cyclopropyl; m is selected from 0, 1 or 2; R2is selected from hydrogen, C 1-3 alkyl; A ring is selected from a benzene ring; a is selected from 0, 1, 2, 3 or 4; R3are each independently selected from the group consisting of hydrogen, halogen, COOR 3a , OCOR 3a , COR 3a , OR 3a , unsubstituted or substituted by one or more than one R 3b C 1-3 alkyl, C 2-3 alkenyl, C 2-3 alkynyl, 3-6 membered saturated cycloalkyl, 3-6 membered saturated nitrogen containing heterocyclyl; R 3a is selected from the group consisting of C 1-3 alkyl, R 3b are each independently selected from the group consisting of halogen, C 1-3 alkyl.

9. The compound, pharmaceutically acceptable salt thereof, stereoisomer thereof, deuterated derivative thereof, or solvate thereof of any one of claims 1-8, wherein, B ring is selected from a 3-6 membered saturated cycloalkyl, a 3-6 membered saturated heterocyclyl; b is selected from 0, 1, 2, 3 or 4; R4 is independently selected from hydrogen, halogens, and COOR. 4a OCOR 4a COR 4a OR 4a Not replaced or replaced by one or more R 4b The following groups are substituted: C 1-3 Alkyl, C 2-3 alkenyl, C 2-3 Alkyne group, 3-6 membered saturated cycloalkyl group, 3-6 membered saturated nitrogen-containing heterocyclic group; R 4a Selected from C 1-3 Alkyl, R 4b Each is independently selected from halogens, C 1-3 alkyl; R5is selected from C 1-3 alkyl.

10. The compound, pharmaceutically acceptable salt thereof, stereoisomer thereof, deuterated form thereof or solvate thereof according to claim 9, characterized in that, the B ring is selected from a 3-membered saturated cycloalkyl, a 4-membered saturated cycloalkyl, a 5-membered saturated cycloalkyl, a 6-membered saturated cycloalkyl.

11. The compound, pharmaceutically acceptable salt thereof, stereoisomer thereof, deuterated derivative thereof, or solvate thereof of claim 10, wherein, the for 12. The compound of claim 1, a pharmaceutically acceptable salt thereof, a stereoisomer thereof, a deuterated form thereof, or a solvate thereof, wherein The compound is one of the following compounds:

13. A pharmaceutical composition, characterized by, The pharmaceutical composition is a preparation prepared by adding a pharmaceutically acceptable adjuvant to the compound, the pharmaceutically acceptable salt thereof, the stereoisomer thereof, the deuterated compound thereof, or the solvate thereof of any one of claims 1-12 as an active ingredient.

14. Use of the compound, the pharmaceutically acceptable salt thereof, the stereoisomer thereof, the deuterated compound thereof, or the solvate thereof of any one of claims 1-12 in the preparation of a TYK2 inhibitor.

15. Use according to claim 14, characterized in that, The TYK2 inhibitor is a drug for preventing and / or treating inflammation, a skin disease, an autoimmune disease, cancer, or a neurodegenerative disease.

16. Use according to claim 15, characterized in that, The autoimmune disease is lupus, rheumatoid arthritis, multiple sclerosis, psoriasis, ulcerative colitis, Crohn's disease, type I diabetes, or Sjogren's syndrome; the skin disease is vitiligo, skin rash, atopic dermatitis, or lichen planus; the cancer is kidney cancer, liver cancer, pancreatic cancer, gastric cancer, breast cancer, prostate cancer, head and neck cancer, thyroid cancer, lung cancer, glioblastoma, melanoma, lymphoma, or leukemia; and the neurodegenerative disease is Alzheimer's disease, Parkinson's disease, Huntington's disease, or amyotrophic lateral sclerosis.

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