Nitrogen-containing cyclic compound, pharmaceutical composition thereof, and use thereof

Abstract

Provided are a nitrogen-containing cyclic compound, a pharmaceutical composition thereof, and a use thereof. Specifically provided are a compound as shown in formula (I), a stereoisomer or a pharmaceutically acceptable salt thereof, and a use thereof in the preparation of a TYK2 inhibitor. This type of compound inhibits TYK2-mediated immune regulation and control functions by means of targeting pseudokinase domain JH2 of TYK2. Therefore, the compound can be used as a TYK2 inhibitor, and also exhibits good pharmacokinetic properties.

Description

Nitrogen-containing cyclic compounds, pharmaceutical compositions and applications thereof

[0001] This application claims the benefit of Chinese patent application No. 2023112332952 filed on September 22, 2023. This application incorporates the entirety of the aforementioned Chinese patent application. Technical Field

[0002] The present invention relates to a nitrogen-containing cyclic compound, a pharmaceutical composition thereof and application thereof. Background Art

[0003] Janus kinases (JAKs) are a family of intracellular non-receptor tyrosine kinases that transduce cytokine-mediated signals through the JAK-STAT pathway. The four members of the JAK family—Janus kinase 1 (JAK1), Janus kinase 2 (JAK2), Janus kinase 3 (JAK3), and tyrosine kinase 2 (TYK2)—have been shown to be key components of cytokine-mediated responses. When cytokine receptors bind to their ligands, they activate receptor-associated JAKs, leading to receptor phosphorylation. The phosphorylated tyrosine sites bind to STAT proteins containing SH2 domains, leading to STAT recruitment to the receptors and phosphorylation by JAKs. Subsequently, phosphotyrosine-mediated STAT dimerization occurs, and the activated STAT dimers translocate to the nucleus and activate the transcription of their target genes, thereby regulating a variety of cell functions, including growth, activation, and differentiation.

[0004] TYK2 is the first subtype of the JAK family to be discovered. It mediates the functions of cytokines such as IFN-α, IL-6, IL-10, IL-12, and IL-23. Studies have shown that TYK2 deletion mutations can effectively inhibit the occurrence of immune diseases such as allergies, autoimmunity, and inflammation. IL-23 plays a crucial role in the development and progression of psoriasis. Recent studies have shown that the pathogenesis of psoriasis is that endogenous unknown antigens activate antigen-presenting cells (APCs) to secrete IL-23. IL-23 activates Th17 cells, which secrete IL-17 and other cytokines, inducing keratinocyte differentiation and division and secretion of IL-23, further stimulating inflammation and keratinocyte proliferation to produce psoriasis. TYK2 and JAK2 jointly mediate the downstream signaling pathway of IL-23. Inhibiting JAK2 can lead to anemia and other blood-related side effects. Therefore, targeting TYK2 to inhibit the IL-23 signaling pathway is a promising strategy for treating psoriasis.

[0005] JAK1, JAK2, JAK3, and TYK2, members of the JAK family of proteins, all possess a JAK homology domain (JH). The JH1 domain is also known as the kinase domain, while JH2 is a pseudokinase domain. Most approved JAK inhibitors are not highly selective, posing safety concerns. This is because these JAK inhibitors all bind to the ATP site of the JH1 domain within the JAK protein, and the JH1 domains of different JAK family members are highly similar. Consequently, when intracellular drug concentrations increase, a JAK inhibitor may also affect ATP binding to other JAK kinases. Although the JH2 domain of TYK2 is very similar to the JH1 domain and contains an ATP binding site very similar to that of the JH1 domain, specific residue differences preclude JH2 from catalytic function. Evidence suggests that small molecule ligands binding to JH2 stabilize the autoinhibitory interaction between the JH2 domain and the JH1 active site. By binding to the JH2 domain of TYK2 and inhibiting TYK2, a high level of selectivity against other JAK family members and the entire kinase family can be maintained. In September 2022, the FDA approved Bristol-Myers Squibb's (BMS) oral selective tyrosine kinase 2 (TYK2) allosteric inhibitor deucravacitinib for the treatment of adult patients with moderate to severe plaque psoriasis who are suitable for systemic treatment or phototherapy.

[0006] The Tyk2-STAT signaling pathway, which is specific for Tyk2, is closely related to a variety of diseases such as psoriasis, systemic lupus erythematosus, and inflammatory bowel disease. Targeting these cytokines with biological agents to intervene in the Tyk2-STAT signaling pathway has been shown to be a viable option for treating autoimmune and inflammatory diseases. Many diseases are associated with abnormal cell responses triggered by kinase-mediated events. These diseases include, but are not limited to, autoimmune diseases, inflammatory diseases, bone diseases, metabolic diseases, nervous system and neurodegenerative diseases, cancer, cardiovascular diseases, allergies and asthma, Alzheimer's disease, and hormone-related diseases. Therefore, there is still a need to find protein kinase inhibitors that can be used as therapeutic agents.

[0007] Summary of the Invention

[0008] The present invention provides novel nitrogen-containing cyclic compounds, pharmaceutical compositions thereof, and uses thereof. These compounds inhibit TYK2-mediated immune regulation by targeting the pseudokinase domain JH2 of TYK2, thereby inhibiting TYK2-mediated immune regulation. These compounds also exhibit favorable pharmacokinetic properties.

[0009] The present invention provides a compound as shown in Formula I, its stereoisomers or pharmaceutically acceptable salts thereof,

[0010] in,

[0011] R 1 is a C1-C6 alkyl group, 1-1 Substituted C1-C6 alkyl, C1-C6 alkoxy, one or more R 1-2 Substituted C1-C6 alkoxy, 3-6 membered cycloalkyl, one or more R 1-3 Substituted 3-6 membered cycloalkyl, 3-7 membered heterocycloalkyl, substituted by one or more R 1-4 Substituted 3-7 membered heterocycloalkyl, -OR 1-5 or by one or more R 1-5-1 Substituted -OR 1-5 , the 3-7 membered heterocycloalkyl group and one or more R 1-4 The heteroatoms in the 3-7-membered heterocycloalkyl group in the substituted 3-7-membered heterocycloalkyl group are selected from one or more of N, O and S, and the number of heteroatoms is 1, 2 or 3;

[0012] R 1-1 and R 1-2 Each is independently halogen, hydroxy or 3-6 membered cycloalkyl;

[0013] R 1-3 Each independently represents halogen, hydroxyl, C1-C6 alkyl, one or more R 1-3-1 Substituted C1-C6 alkyl, C1-C6 alkoxy or one or more R 1-3-2 Substituted C1-C6 alkoxy;

[0014] R 1-3-1 and R 1-3-2 each independently deuterium or halogen;

[0015] R 1-4 Each independently represents halogen, hydroxyl, C1-C6 alkyl, one or more R 1-4-1 Substituted C1-C6 alkyl, C1-C6 alkoxy or one or more R 1-4-2 Substituted C1-C6 alkoxy;

[0016] R 1-4-1 and R 1-4-2 each independently deuterium or halogen;

[0017] R 1-5 is a 3-6 membered cycloalkyl group;

[0018] R 1-5-1 Each is independently halogen, hydroxy or C1-C6 alkyl;

[0019] Ring A is a 4-12 membered heterocyclic ring or a 5-10 membered heteroaromatic ring, wherein the heteroatoms in the 4-12 membered heterocyclic ring and the 5-10 membered heteroaromatic ring include at least one nitrogen atom, and the remaining heteroatoms are selected from one or more of N, O and S, and the number of heteroatoms is 1, 2 or 3;

[0020] R 2 Each is independently halogen, hydroxyl, cyano, oxo (=O), C1-C6 alkyl, 2-1 Substituted C1-C6 alkyl, C1-C6 alkoxy or one or more R 2-2 Substituted C1-C6 alkoxy;

[0021] R 2-1 and R 2-2 are each independently halogen or hydroxy;

[0022] R 3 is a C1-C6 alkyl group, 1-1 Substituted C1-C6 alkyl, C1-C6 alkoxy, one or more R 1-2 Substituted C1-C6 alkoxy, 3-6 membered cycloalkyl, one or more R 1-3 Substituted 3-6 membered cycloalkyl, 3-7 membered heterocycloalkyl, substituted by one or more R 1-4 Substituted 3-7 membered heterocycloalkyl, -OR 1-5 or by one or more R 1-5-1 Substituted -OR 1-5 , the 3-7 membered heterocycloalkyl group and one or more R 1-4 The heteroatoms in the 3-7-membered heterocycloalkyl group in the substituted 3-7-membered heterocycloalkyl group are selected from one or more of N, O and S, and the number of heteroatoms is 1, 2 or 3;

[0023] R 4 is hydrogen, halogen, hydroxy, cyano, oxo, C1-C6 alkyl, 1-1 Substituted C1-C6 alkyl, C1-C6 alkoxy, one or more R 1-2 Substituted C1-C6 alkoxy, 3-6 membered cycloalkyl, one or more R 1-3 Substituted 3-6 membered cycloalkyl, 3-7 membered heterocycloalkyl, substituted by one or more R 1-4 Substituted 3-7 membered heterocycloalkyl, -OR 1-5 or by one or more R 1-5-1 Substituted -OR 1-5 , the 3-7 membered heterocycloalkyl group and one or more R 1-4 The heteroatoms in the 3-7-membered heterocycloalkyl group in the substituted 3-7-membered heterocycloalkyl group are selected from one or more of N, O and S, and the number of heteroatoms is 1, 2 or 3;

[0024] m is 0, 1, 2 or 3.

[0025] In a certain embodiment, in the compound represented by Formula I, its stereoisomers or pharmaceutically acceptable salts thereof, certain groups have the following definitions, and the definitions of the unmentioned groups are as described in any embodiment of the present invention (this paragraph is hereinafter referred to as "in a certain embodiment").

[0026] In one embodiment, each C1-C6 alkyl group can be independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentane, isopentane or neopentane, for example, methyl, ethyl, isobutyl or neopentane.

[0027] In a certain embodiment, each C1-C6 alkoxy group can be independently a C1-C4 alkoxy group, and can also be a methoxy group, an ethoxy group, an n-propoxy group, an isopropoxy group, an n-butoxy group, an isobutoxy group, a sec-butoxy group or a tert-butoxy group, for example, a methoxy group, an ethoxy group or an isopropoxy group.

[0028] In a certain embodiment, each 3-6 membered cycloalkyl group can be independently a 3-6 membered monocyclic cycloalkyl group, and can be a cyclopropyl group, a cyclobutyl group, a cyclopentyl group or a cyclohexyl group, for example, a cyclopropyl group or a cyclobutyl group.

[0029] In a certain embodiment, each 3-7 membered heterocycloalkyl group may be independently a 3-7 membered monocyclic heterocycloalkyl group, or a 3-7 membered monocyclic heterocycloalkyl group having one or two heteroatoms selected from N and O.

[0030] In one embodiment, each halogen can independently be fluorine, chlorine, bromine or iodine, for example fluorine or chlorine.

[0031] In a certain embodiment, each 4-12 membered heterocycle may be a 4-12 membered monocyclic heterocycle or a 4-12 membered polycyclic heterocycle (e.g., spirocycle); the 4-12 membered monocyclic heterocycle may be a 4-6 membered monocyclic heterocycle, preferably a 4-6 membered monocyclic heterocycle wherein the heteroatoms are selected from 1 or 2 of N and O, for example The 4-12 membered polycyclic heterocycle is preferably

[0032] In a certain embodiment, the heteroatom in each 5-10 membered heteroaromatic ring is preferably N, and the number of heteroatoms is preferably 1 or 2.

[0033] In one embodiment, each 5-10 membered heteroaromatic ring may be a 5-6 membered heteroaromatic ring, preferably a 5-6 membered heteroaromatic ring having 1 or 2 heteroatoms selected from N, for example

[0034] In one scenario, R 1 is a C1-C6 alkyl group, 1-1Substituted C1-C6 alkyl, C1-C6 alkoxy, one or more R 1-2 Substituted C1-C6 alkoxy, 3-6 membered cycloalkyl, one or more R 1-3 Substituted 3-6 membered cycloalkyl or -OR 1-5 Preferably, R 1 is a C1-C6 alkyl group, 1-1 Substituted C1-C6 alkyl, C1-C6 alkoxy, one or more R 1-2 Substituted C1-C6 alkoxy, one or more R 1-3 Substituted 3-6 membered cycloalkyl or -OR 1-5 More preferably, R 1 is a C1-C6 alkoxy group, one or more R 1-2 Substituted C1-C6 alkoxy or -OR 1-5 .

[0035] In one scenario, R 1-1 are each independently halogen or hydroxy.

[0036] In one scenario, R 1-2 Each is independently fluorine, hydroxy or 3-6 membered cycloalkyl.

[0037] In one scenario, R 1-3 Each independently represents halogen, hydroxy, C1-C6 alkyl, C1-C4 alkoxy or is replaced by one or more R 1-3-2 Substituted C1-C4 alkoxy; preferably halogen, hydroxy, C1-C4 alkoxy or one or more R 1-3-2 Substituted C1-C4 alkoxy.

[0038] In one scenario, R 1-3-2 Each is independently deuterium or fluorine, preferably deuterium.

[0039] In a certain embodiment, ring A is a 4-6 membered monocyclic heterocycle or a 5-6 membered heteroaromatic ring, and the heteroatoms in the 4-6 membered monocyclic heterocycle and the 5-6 membered heteroaromatic ring are selected from 1 or 2 of N and O; preferably, ring A is a 6 membered monocyclic heterocycle or a 5 membered heteroaromatic ring, and the heteroatoms in the 6 membered monocyclic heterocycle are selected from 1 or 2 of N and O, and the heteroatoms in the 5 membered heteroaromatic ring are selected from N, and the number of heteroatoms is 1 or 2.

[0040] In one scenario, R 2 Each independently represents halogen, hydroxyl, C1-C6 alkyl, one or more R 2-1 Substituted C1-C6 alkyl, or C1-C6 alkoxy; preferably halogen or hydroxy.

[0041] In one scenario, R 2-1are each independently halogen.

[0042] In one scenario, R 3 It is a C1-C6 alkyl group.

[0043] In one scenario, R 4 For hydrogen.

[0044] In one embodiment, m is 0, 1 or 2.

[0045] In one plan, for

[0046] In one scenario, R 1 for

[0047] In one scenario, R 2 is hydroxy, fluorine, chlorine, methyl, methoxy or trifluoromethyl.

[0048] In one embodiment, Ring A is

[0049] In one plan, for

[0050] In one embodiment, the compound as shown in Formula I is any of the following compounds:

[0051] The present invention provides a pharmaceutical composition comprising:

[0052] (1) (a therapeutically effective amount of) substance A, wherein substance A is a compound of formula I as described above, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, and

[0053] (2) Pharmaceutically acceptable excipients.

[0054] The present invention also provides the use of the compound represented by Formula I, its stereoisomers, or pharmaceutically acceptable salts thereof, or the pharmaceutical composition described above, in the preparation of a TYK2 inhibitor. In such applications, the TYK2 inhibitor can be used in mammals in vivo or in vitro, primarily for experimental purposes, for example, as a standard or control sample for comparison, or prepared into a kit according to conventional methods in the art to provide rapid detection of TYK2 inhibition effects.

[0055] The present invention also provides the use of the compound of Formula I, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described above, in the preparation of a medicament for treating a disease mediated by TYK2. The disease may be an autoimmune disease, an inflammatory disease, a bone disease, a metabolic disease, a neurodegenerative disease, cancer, a cardiovascular disease, an allergy, asthma, or Alzheimer's disease.

[0056] In addition to the foregoing, when used in the specification and claims of this application, unless otherwise specifically indicated, the following terms have the following meanings:

[0057] As used herein, the compounds shown in Formula I of the present invention may contain one or more chiral centers or chiral axes and exist in different optically active forms. When a compound contains one chiral center, the compound comprises enantiomers. The present invention includes both isomers and mixtures of isomers, such as racemic mixtures. Enantiomers can be resolved by methods known in the art, such as crystallization and chiral chromatography. When the compound shown in Formula I contains more than one chiral center, diastereomers may exist. The present invention includes resolved optically pure specific isomers and mixtures of diastereomers. Diastereomers can be resolved by methods known in the art, such as crystallization and chiral chromatography.

[0058] The term "stereoisomer" includes conformational isomers and configurational isomers, wherein configurational isomers mainly include cis-trans isomers and optical isomers. The compounds described in the present invention may exist in the form of stereoisomers, and therefore encompass all possible stereoisomeric forms, including but not limited to cis-trans isomers, enantiomers, diastereomers, atropisomers, etc. The compounds described in the present invention may also exist in the form of any combination or any mixture of the aforementioned stereoisomers, such as meso-, racemic-, and equal mixtures of atropisomers, or single enantiomers, single diastereomers, or mixtures thereof, or single atropisomers or mixtures thereof.

[0059] The term "pharmaceutically acceptable salt" refers to salts prepared from compounds of the present invention with relatively nontoxic, pharmaceutically acceptable acids or bases. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of a pharmaceutically acceptable base in neat solution or in a suitable inert solvent. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of a pharmaceutically acceptable acid in neat solution or in a suitable inert solvent.

[0060] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0061] The term "alkyl" refers to a group having the specified number of carbon atoms (e.g., C 1-6 or C1-C6), linear or branched, saturated monovalent hydrocarbon group. Alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, and the like.

[0062] The term "alkoxy" refers to the group -O-alkyl, wherein alkyl is as defined above.

[0063] The term "cycloalkyl" refers to a saturated monocyclic, spirocyclic, bridged, or paracyclic cyclic group having a specified number of ring carbon atoms (e.g., 3-6 members), wherein the ring atoms consist solely of carbon atoms. Cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0064] The term "heterocycloalkyl" or "heterocycle" refers to a cyclic group having a specified number of ring atoms (e.g., 3-7 members or 4-12 members), a specified number of heteroatoms (e.g., 1, 2, or 3), and a specified type of heteroatom (e.g., one, two, or more of N, O, and S), which can be monocyclic or bicyclic (e.g., cyclic or spirocyclic). Heterocycloalkyl or heterocycle includes, but is not limited to, oxetane, azetidinyl, tetrahydropyrrolyl, piperazinyl, piperidinyl, morpholinyl, and the like.

[0065] The term "heteroaromatic ring" refers to a cyclic group having a specified number of ring atoms (e.g., 5-10 or 5-6 members), a specified number of heteroatoms (e.g., 1, 2, or 3), and a specified heteroatom species (e.g., one, two, or more of N, O, and S). It can be a monocyclic or bicyclic ring (e.g., a cyclic ring or a spirocyclic ring), and at least one ring has aromaticity (in accordance with Huckel's rule). The heteroaryl group is connected to other fragments in the molecule through a ring having aromaticity or a ring that does not have aromaticity. Heteroaromatic rings include, but are not limited to, furan, pyrrole, thiophene, pyrazole, imidazole, oxazole, thiazole, pyridine, pyrimidine, and the like.

[0066] The term "one or more" means 1, 2, 3, 4, 5, 6, 7, 8, 9 or more.

[0067] It will be understood by those skilled in the art that the structural formulas used in the present invention to describe groups are based on the conventions used in the art. It means that the corresponding group R is connected to other fragments and groups in the compound through this site.

[0068] The term "pharmaceutically acceptable excipients" refers to excipients and additives used in the production of pharmaceuticals and the preparation of prescriptions. These excipients are all substances, other than the active ingredient, contained in a pharmaceutical preparation. For more information, see Part IV of the Pharmacopoeia of the People's Republic of China (2020 Edition) or the Handbook of Pharmaceutical Excipients (Raymond C. Rowe, 2009 Sixth Edition).

[0069] In the present invention, a "pharmaceutical composition" refers to a formulation comprising a compound of the present invention and a medium generally accepted in the art for delivering a biologically active compound to a mammal (e.g., a human). The medium includes a pharmaceutically acceptable carrier. The purpose of a pharmaceutical composition is to facilitate administration to an organism, thereby facilitating absorption of the active ingredient and thereby exerting its biological activity.

[0070] The pharmaceutical compositions of the present invention can be prepared according to the disclosed content using any method known to those skilled in the art, such as conventional mixing, dissolving, granulating, emulsifying, grinding, encapsulating, embedding or lyophilizing processes.

[0071] The term "treat" refers to therapeutic treatment. When referring to a specific condition, treatment means: (1) alleviating the disease or one or more biological manifestations of the condition, (2) interfering with (a) one or more points in the biological cascade that leads to or causes the condition or (b) one or more biological manifestations of the condition, (3) ameliorating one or more symptoms, effects, or side effects associated with the condition or one or more symptoms, effects, or side effects associated with the condition or its treatment, or (4) slowing the progression of the condition or one or more biological manifestations of the condition.

[0072] The term "prevent" refers to the reduction of the risk of acquiring or developing a disease or disorder.

[0073] The term "therapeutically effective amount" refers to an amount administered to a patient that is sufficient to effectively treat a disease. The therapeutically effective amount will vary depending on the type of compound, the type of disease, the severity of the disease, the age of the patient, etc., but can be adjusted by those skilled in the art as appropriate.

[0074] The term "patient" refers to any animal, preferably a mammal, and most preferably a human, that is about to receive or has received a compound or composition according to embodiments of the present invention. The term "mammal" includes any mammal. Examples of mammals include, but are not limited to, cattle, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, and humans, with humans being the most preferred.

[0075] Without violating the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain preferred embodiments of the present invention.

[0076] The reagents and raw materials used in the present invention are commercially available.

[0077] The positive progress of the present invention is that the compounds of the present invention have good inhibitory activity against pSTAT5 and can therefore be used as TYK2 inhibitors while showing good pharmacokinetic properties. DETAILED DESCRIPTION

[0078] When the preparation route is not included, the raw materials and reagents used in the present invention are all known products, which can be synthesized according to methods known in the art, or can be obtained by purchasing commercial products. No further purification is required for the commercially available reagents used.

[0079] Room temperature refers to 20-30℃.

[0080] Unless otherwise specified in the reaction examples, all reactions were carried out under a nitrogen atmosphere, which means that the reaction flask was connected to a nitrogen balloon of approximately 1 L.

[0081] The hydrogenation reaction is usually carried out by evacuating the flask and filling it with hydrogen, and this operation is repeated three times. The hydrogen atmosphere means that the reaction flask is connected to a hydrogen balloon of about 1L.

[0082] Microwave reaction use Initiator + microwave reactor.

[0083] The structures of the compounds of the present invention were determined by nuclear magnetic resonance (NMR) and mass spectrometry (MS). -6 The units of ppm are given. NMR measurements were performed using a BRUKER AVANCE III, 400M NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and deuterated methanol (CD3OD), with tetramethylsilane (TMS) as the internal standard. The following abbreviations are used for NMR signal multiplicities: s = singlet, brs = broad, d = doublet, t = triplet, and m = multiplet. Coupling constants are listed as J values ​​and are measured in Hz.

[0084] Rapid column chromatography used an Aeger (FS-9200T) automatic column machine, and silica gel pre-packed columns used Santai Pre-packed columns. Thin layer chromatography silica gel plates use Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates. The specifications used for thin layer chromatography separation and purification products are 0.4mm to 0.5mm.

[0085] Reverse phase preparative chromatography was performed using a Waters-3767 reverse phase preparative chromatograph.

[0086] Preparative column: Waters SunFire, 250*19mm, 5mm or Xbridge Prep C18 or Gemini C18 21.2*250 5μm.

[0087] Mobile phase: A: 0.05% ammonia-water solution; B: acetonitrile.

[0088] Detection wavelengths: 254nm and 214nm.

[0089] Flow rate: 20mL / min.

[0090] LCMS was performed using a Waters ARC HPLC-QDA instrument.

[0091] Chromatographic column: ACQUITY UPLC BEH C18 3.5μm 3.0*50mm.

[0092] Ion source: ESI.

[0093] Mobile phase: A: 0.05% ammonia-water solution; B: 0.05% ammonia-acetonitrile solution.

[0094] Detection wavelengths: 254nm and 214nm.

[0095] Flow rate: 1.5 mL / min; run time: 3.5 min.

[0096] Waters W2489 Instrument was used for HPLC.

[0097] Chromatographic column: Xbridge C18, 4.6*50mm.

[0098] Mobile phase: A: 0.1% ammonia-water solution; B: acetonitrile.

[0099] Detection wavelengths: 254nm and 214nm.

[0100] Running time: 9.0min.

[0101] Supercritical fluid chromatography (SFC) was performed using a Waters SFC 150 instrument.

[0102] Column: DAICEL or DAICEL

[0103] Mobile phase: A: Supercritical CO2; B: 0.05% ammonia-methanol solution

[0104] Detection wavelength: 254nm and 214nm

[0105] The synthetic methods of some intermediates in the invention are as follows:

[0106] Intermediate 1

[0107] Intermediate 1 was prepared by the following steps:

[0108] Step A: Sodium ethoxide (2.6 g) was added to an ethanol solution (60 mL) of ethyl 5-amino-1H-pyrazole-4-carboxylate (Int-1a, 2 g) and diethyl malonate (Int-1b, 4.1 g). The mixture was reacted at 100°C for 16 hours. LC / MS showed that the reaction was complete. The system was spin-dried and poured into 120 mL of water. The aqueous phase was adjusted to pH 3-4 with concentrated hydrochloric acid, and a solid precipitated. The solid was collected by filtration and washed twice with water, then dried in vacuo to give Int-1c (2.2 g) as a yellow solid. LC / MS (ESI + )m / z:224.1[M+H] + .

[0109] Step B: To a solution of Int-1c (24.5 g) in phosphorus oxychloride (100 mL) was slowly added dropwise N,N-diethylaniline (24.6 g) and the reaction was carried out at 100°C for 4 hours. TLC indicated that the reaction was complete. The phosphorus oxychloride was evaporated under reduced pressure, and the residue was slowly added to ice water (600 mL). The aqueous phase was extracted three times with dichloromethane (DCM). The organic phases were combined and washed twice with saturated Na2CO3 and once with saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified on a silica gel column (petroleum ether / ethyl acetate = 8 / 1 to 1 / 1, v / v) to obtain Int-1d (15.9 g, 55% yield) as a yellow solid.

[0110] Step C: To a mixed solution of Int-1d (15.9 g) in acetonitrile (160 mL) and water (4 mL) were added potassium carbonate (25.3 g) and methylamine hydrochloride (5.4 g). Stirred at room temperature for 1 hour, LC / MS showed that the reaction was complete. The reaction solution was concentrated under reduced pressure, water (200 mL) was added and stirred at room temperature for 10 minutes. Filtered, the filter cake was rinsed with water and dried under vacuum to obtain a yellow solid Int-1e (14.5 g, yield: 93%). LC / MS (ESI + )m / z:255.0(M+H) + .

[0111] Step D: To a solution of Int-1e (14.5 g) in dichloromethane (150 mL) were added tert-butyl tert-butyloxycarbonyl carbonate (13.1 g) and 4-dimethylaminopyridine (287.8 mg). The mixture was stirred at room temperature for 2 hours. TLC indicated the reaction was complete. The reaction solution was concentrated, and the residue was purified on a silica gel column (petroleum ether / ethyl acetate = 8 / 1 to 2 / 1, v / v) to afford Int-1f (14 g, 67% yield) as a yellow solid.

[0112] Step E: To a solution of Int-1f (2 g) in toluene (20 mL) was added tributyltin oxide (4.4 g). The mixture was stirred at 125°C for 16 hours. LC / MS showed that the reaction was complete. The reaction solution was concentrated, and the residue was added with a saturated aqueous sodium carbonate solution (40 mL) and stirred at room temperature for 10 minutes. The aqueous phase was washed twice with methyl tert-butyl ether, and then the aqueous phase was adjusted to pH = 3-4 with concentrated hydrochloric acid and extracted three times with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to give Int-1g (1.2 g, yield 65%) as a yellow solid. LC / MS (ESI + )m / z:348.9(M+Na) + .

[0113] Step F: To a dichloromethane solution (10 mL) of Int-1g (1 g) were added (1R,2S)-2-fluorocyclopropaneaminomethylbenzenesulfonate (1 g), N,N-diisopropylethylamine (1.38 g) and T3P (50% in ethyl acetate) (1 g). After stirring at room temperature for 16 hours, the reaction solution was concentrated and the residue was purified by silica gel column (petroleum ether / ethyl acetate = 8 / 1 to 1 / 1, V / V) to give Int-1 (520 mg, yield: 44%) as a yellow solid. LC / MS (ESI + )m / z:384.0[M+H] + .

[0114] Intermediate 2

[0115] Intermediate 2 was prepared by the following steps:

[0116] Step A: To a solution of methyl 2-oxo-1H-pyridine-3-carboxylate (Int-2a, 500 mg, 3.27 mmol) in N,N-dimethylformamide (10 mL) was added O-diphenylphosphoryl hydroxylamine (913.71 mg, 3.92 mmol) and cesium carbonate (2.66 g, 8.16 mmol) and stirred at room temperature for 30 min. The reaction mixture was concentrated and the residue was washed with dichloromethane and filtered. The organics were concentrated and purified on a silica gel column (dichloromethane / methanol = 99 / 1 to 95 / 5, V / V) to give Int-2 (300 mg, 1.78 mmol, yield: 54%). 1 H NMR (400MHz, DMSO-d6) δ8.07(dd,J=6.4,2.4Hz,1H),7.98(dd,J=6.4,2.4Hz,1H),6.34(s,2H),6.32–6.28(m,1H),3.75(s,3H).

[0117] Intermediate 3

[0118] Intermediate 3 was prepared by the following steps:

[0119] Step A: 3-Aminopyridin-2(1H)one (Int-3a, 25 g) and di-tert-butyl dicarbonate (99 g) were dissolved in methanol (MeOH, 200 mL). The reaction mixture was reacted at 40°C for 16 hours. The reaction solution was concentrated and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 0% to 35%, V / V) to give Int-3b (37 g) as a green solid. LC / MS (ESI + )m / z:211.1[M+H] + .

[0120] Step B: Int-3b (13 g), (aminooxy)diphenylphosphine oxide (18.8 g) and Cs2CO3 (44.3 g) were added to N,N-dimethylformamide (DMF, 120 mL) and reacted at room temperature for 25 minutes. The reaction mixture was filtered and the filtrate was collected and concentrated. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 0% to 60%, V / V) to give Int-3 (10.4 g) as a green solid. LC / MS (ESI + )m / z:226.1[M+H] + .

[0121] The synthesis method of the embodiment compounds of the present invention is as follows:

[0122] Example 1

[0123] N-((1R,2S)-2-fluorocyclopropyl)-7-(methylamino)-5-((2-carbonyl-1-(piperidin-1-yl)-1,2-dihydropyridin-3-yl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxamide (Compound 1)

[0124] Compound 1 was prepared by the following steps:

[0125] Step A: Dissolve 1,1,3,3-tetramethoxypropane (1a, 9.32 g) and zinc chloride (258 mg) in acetic anhydride (30 mL). Then slowly add dimethyl malonate (1b, 5 g) dropwise to the reaction mixture. Reaction was continued at 140°C for 4 hours. LC / MS indicated the reaction was complete. The reaction mixture was concentrated to dryness, and the residue was purified by column chromatography (petroleum ether / ethyl acetate = 10 / 1) to afford 1c (5 g), a yellow liquid. 1HNMR (400MHz, DMSO-d6) δ7.43(d,J=12.1Hz,1H),7.09(d,J=12.3Hz,1H),6.23(t,J=12.2Hz,1H),3.80(s,3H),3.74(d,J=4.0Hz,6H).

[0126] Step B: Piperidin-1-amine (1d, 500 mg) and potassium carbonate (1.38 g) were dissolved in ethanol (10 mL), and then compound 1c (1 g) was added at 0°C. The reaction was allowed to proceed at room temperature for 4 hours. LC / MS showed that the reaction was complete. The reaction solution was concentrated, and the residue was then purified by column chromatography (petroleum ether / ethyl acetate = 3 / 1) to give a light yellow liquid 1e (250 mg). LC / MS (ESI + )m / z:237.1[M+H] + .

[0127] Step C: Compound 1e (250 mg) and lithium hydroxide monohydrate (100 mg) were dissolved in a mixed solution of tetrahydrofuran (2 mL), methanol (2 mL) and water (1 mL). The mixture was reacted at room temperature for 4 hours. LC / MS showed that the reaction was complete. The reaction solution was concentrated and the residue was purified by reverse phase column (formic acid system) to obtain a brown solid 1f (120 mg). LC / MS (ESI + )m / z:223.1[M+H] + .

[0128] Step D: Compound 1f (120 mg), diphenylphosphoryl azide (210 mg), and triethylamine (80 mg) were dissolved in a mixed solution of toluene (5 mL) and tert-butanol (3 mL). The mixture was reacted at 100°C for 2 hours. LC / MS showed that the reaction was complete. The reaction solution was concentrated, and the residue was purified by reverse phase column (formic acid system) to obtain 1 g (90 mg) of a yellow solid. LC / MS (ESI + )m / z:237.9[M+H] + .

[0129] Step E: Dissolve 1g (90mg) in 4M hydrochloric acid / dioxane solution (5mL). React at 50°C for 6 hours. LC / MS shows the reaction is complete. Concentrate the reaction solution to obtain a yellow oil 1h (50mg). LC / MS (ESI + )m / z:193.98[M+H] + .

[0130] Step F: Compound 1h (50 mg), Int-1 (80 mg), cesium carbonate (170 mg), 1,1'-binaphthyl-2,2'-bis(diphenylphosphine) (20 mg) and palladium acetate (10 mg) were dissolved in dioxane (2 mL). After nitrogen was replaced in the reaction system, the reaction was continued at 70°C for 3 hours. LC / MS showed that the reaction was complete. The reaction solution was concentrated and the residue was purified by reverse phase column (formic acid system) to obtain 1i (30 mg) as a white solid. LC / MS (ESI + )m / z:541.1[M+H] + .

[0131] Step G: Compound 1i (30 mg) was dissolved in 4 M hydrochloric acid / dioxane solution (5 mL). The mixture was allowed to react at room temperature for 2 hours. LC / MS showed that the reaction was complete. The reaction solution was concentrated and the residue was purified by reverse phase column (formic acid system) to obtain a light yellow solid, compound 1 (4.1 mg). LC / MS (ESI + )m / z:441.2[M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ8.86(s,1H),8.23(s,1H),8.10(dd,J=7.4,1.7Hz,1H),7.91(d,J=5.2Hz,1H),7.78(d,J=4.7Hz,1H),7.29(dd,J=7.1,1.8H z,1H),6.31–6.08(m,2H),4.98–4.78(m,1H),2.99(dd,J=9.3,4.8Hz,1H) ,2.91(d,J=4.7Hz,3H),1.67(s,4H),1.53–1.13(m,4H),0.92–0.8(m,1H).

[0132] Example 2

[0133] N-((1R,2S)-2-fluorocyclopropyl)-7-(methylamino)-5-((2-carbonyl-1-(1H-pyrrol-1-yl)-1,2-dihydropyridin-3-yl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxamide (Compound 2)

[0134] Compound 2 was prepared by the following steps:

[0135] Step A: Int-2 (500 mg, 2.97 mmol) was dissolved in acetic acid solution (5 mL), and 2,5-dimethoxytetrahydrofuran (471 mg, 3.57 mmol) was added. The reaction mixture was reacted in an oil bath at 100°C for 3 hours. After the reaction was completed, it was concentrated under reduced pressure, saturated aqueous sodium carbonate solution (3 mL) was added, and the mixture was extracted twice with dichloromethane (4 mL). The organic phase was collected, dried over anhydrous sodium sulfate, filtered, concentrated, and dried to give compound 2b (630 mg, 97% yield).

[0136] Step B: Compound 2b (650 mg, 2.98 mmol) was dissolved in a mixture of methanol (6 mL) and water (6 mL). Lithium hydroxide monohydrate (875 mg, 20.85 mmol) was added and stirred at 25°C for 2 hours. After completion of the reaction, the reaction mixture was concentrated, water was added, and the mixture was extracted three times with dichloromethane. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to afford compound 2c (530 mg, 87% yield).

[0137] Step C: Compound 2c (30 mg, 146 μmol) was dissolved in toluene (1 mL), and diphenylphosphoryl azide (DPPA, 60 mg, 220 μmol) and triethylamine (TEA, 22 mg, 220 μmol) were added. The reaction mixture was stirred at 100°C for 2 hours. After completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 20 / 1) to afford compound 2d (25 mg, 61% yield).

[0138] Step D: Dissolve compound 2d (25 mg, 90 μmol) in dichloromethane (1.2 mL) and add trifluoroacetic acid (0.4 mL). Stir the reaction at 25°C for 1 hour. After completion of the reaction, concentrate the reaction mixture to obtain compound 2e (20 mg, yield: 80%).

[0139] Step E: Compound 2e (20 mg, 114 μmol) was dissolved in dichloromethane (3 mL), and Pd(OAc)2 (2.5 mg, 11 μmol), 1,1'-binaphthyl-2,2'-bis(diphenylphosphine) (BINAP, 14.2 mg, 22 μmol), and Cs2CO3 (111.5 mg, 342 μmol) were added. The reaction system was purged with nitrogen and stirred at 60°C for 1 hour. LCMS indicated the reaction was complete. The reaction solution was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane / methanol = 20 / 1) to afford compound 2f (33 mg, 55% yield).

[0140] Step F: Compound 2f (33 mg, 63 μmol) was dissolved in dichloromethane solution (1 mL) and trifluoroacetic acid (1 mL) was added. The mixed solution was stirred at room temperature for 1 h. LCMS showed that the reaction was complete. The reaction solution was concentrated and the residue was purified by reverse HPLC. Compound 2 (6.2 mg, yield 23%) was obtained. LC / MS (ESI + )m / z:423.1[M+H] + ; 1 HNMR(400MHz,DMSO-d6)δ9.00(s,1H),8.24(d,J=8.0Hz,2H),7.95(d,J=4.8H z,1H),7.79(d,J=4.8Hz,1H),7.45(d,J=8.0Hz,1H),7.08(s,2H),6.32(t,J=7 .2Hz,1H),6.25(s,1H),6.20(s,2H),4.99–4.78(m,1H),3.00(dd,J=9.4,4.6H z,1H),2.90(d,J=4.8Hz,3H),1.23(dq,J=14.8,6.8Hz,H),0.95–0.83(m,1H).

[0141] Example 3

[0142] N-((1R,2S)-2-fluorocyclopropyl)-7-(methylamino)-5-((1-morpholino-2-carbonyl-1,2-dihydropyridin-3-yl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxamide (Compound 3)

[0143] Compound 3 was prepared by the following steps:

[0144] Step A: Morpholin-4-amine (1.2 g) and potassium carbonate (2.76 g) were dissolved in ethanol (20 mL), and compound 1c (2 g) was slowly added dropwise to the reaction solution. The reaction was allowed to proceed at room temperature for 8 hours. LC / MS showed that the reaction was complete. The reaction solution was concentrated, and the residue was purified by column chromatography (petroleum ether / ethyl acetate = 1 / 4) to give 3b (580 mg) as a yellow solid. LC / MS (ESI + )m / z:239.1[M+H] + .

[0145] Step B: Compound 3b (500 mg) and lithium hydroxide monohydrate (310 mg) were dissolved in a mixture of methanol (2 mL), tetrahydrofuran (2 mL) and water (1 mL). The mixture was reacted at room temperature for 1 hour. LC / MS showed that the reaction was complete. The reaction solution was concentrated and the residue was purified by column chromatography (petroleum ether / ethyl acetate = 2 / 1) to obtain 3c (540 mg) as a yellow solid. LC / MS (ESI + )m / z:224.9[M+H] + .

[0146] Step C: Compound 3c (540 mg), DPPA (diphenylphosphoryl azide) (994 mg) and triethylamine (366 mg) were dissolved in a mixed solution of toluene (10 mL) and tert-butanol (3 mL). The mixture was reacted at 100°C for 2 hours. LC / MS showed that the reaction was complete. The reaction solution was concentrated and the residue was purified by column chromatography (petroleum ether / ethyl acetate = 2 / 1) to give 3d (580 mg) as a yellow liquid. LC / MS (ESI + )m / z:240.13(M+H) + .

[0147] Step D: Compound 3d (580 mg) was dissolved in 4M hydrochloric acid / dioxane solution (10 mL). The mixture was reacted at 50°C for 6 hours. LC / MS showed that the reaction was complete. The reaction solution was concentrated and the residue was purified by column chromatography (petroleum ether / ethyl acetate = 1 / 2) to give 3e (110 mg) as a white solid. LC / MS (ESI + )m / z:196.17[M+H] + .

[0148] Step E: Compound 3e (50 mg), Int-1 (80 mg), sodium carbonate (30 mg), 1,1'-binaphthyl-2,2'-bis(diphenylphosphine) (20 mg) and palladium acetate (10 mg) were dissolved in dioxane (2 mL). After nitrogen was replaced in the reaction system, the reaction was incubated at 60 degrees Celsius for 8 hours. LC / MS showed that the reaction was complete. The reaction solution was concentrated to dryness, and the residue was purified by column chromatography (petroleum ether / ethyl acetate = 3 / 1) to obtain a yellow solid 3f (100 mg). LC / MS (ESI + )m / z:543.17[M+H] + .

[0149] Step F: Compound 3f (100 mg) was dissolved in 4 M hydrochloric acid / dioxane solution (5 mL). The mixture was allowed to react at room temperature for 2 hours. LC / MS showed that the reaction was complete. The reaction solution was concentrated and the residue was purified by reverse phase preparative HPLC (formic acid system) to obtain a light gray solid, compound 3 (27.9 mg). LC / MS (ESI+ )m / z:443.2[M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ8.90 (s, 1H), 8.23 ​​(s, 1H), 8.12 (dd, J = 7.4, 1.5Hz, 1H), 7.91(q,J=4.7Hz,1H),7.77(d,J=4.7Hz,1H),7.31(dd,J=7.1,1.7Hz,1H),6.24( s,1H),6.20(t,J=7.2Hz,1H),4.87(dtd,J=65.0,5.8,2.9Hz,1H),3.73(s,8H), 3.04–2.93(m,1H),2.90(d,J=4.9Hz,3H),1.31–1.14(m,1H),0.95–0.78(m,1H).

[0150] Example 4

[0151] 5-((1-(1H-imidazol-1-yl)-2-carbonyl-1,2-dihydropyridin-3-yl)amino)-N-((1R,2S)-2-fluorocyclopropyl)-7-(methylamino)pyrazolo[1,5-a]pyrimidine-3-carboxamide (Compound 4)

[0152] Compound 4 was prepared by the following steps:

[0153] Step A: Dissolve Int-3 (500 mg) in methanol (10 mL) and add paraformaldehyde (67 mg), ammonium chloride (119 mg), and glyoxal (129 mg). Stir at 80°C for 1 hour, return to room temperature, add phosphoric acid (0.1 mL), and then heat to reflux for 16 hours. LC / MS indicated completion of the reaction. The reaction solution was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane / methanol = 97 / 3) to afford compound 4a (70 mg, 11% yield).

[0154] Step B: Compound 4a (50 mg) was dissolved in dichloromethane (3 mL) and trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 1.5 hours. LC / MS showed that the reaction was complete. The reaction mixture was concentrated to give compound 4b (30 mg, yield 94%).

[0155] Step C: Add Int-1 (52 mg), 4b (30 mg), sodium carbonate (54 mg), 1,1'-binaphthyl-2,2'-bis(diphenylphosphine) (42 mg), and palladium acetate (8 mg) to 1,4-dioxane (5 mL). Heat to 60°C and stir overnight under nitrogen. LC / MS indicated completion of the reaction. The reaction mixture was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to afford compound 4c (30 mg, 33% yield).

[0156] Step D: Compound 4c (25 mg) was dissolved in dichloromethane (1 mL) and trifluoroacetic acid (0.3 ml) was added. The reaction mixture was stirred at room temperature for 1.5 hours. LC / MS showed that the reaction was complete. After the reaction mixture was concentrated, the residue was purified by reverse phase preparative HPLC to give compound 4 (8.6 mg, yield 43%). LC / MS (ESI + )m / z:422.2[M+H] + ; 1 H NMR(400MHz,DMSO-d6)δ9.05(s, 1H),8.27(d,J=9.7Hz,2H),8.08(s,1H),7.96(d,J=5.0Hz,1H),7.78(d,J=4.6 Hz,1H),7.61(s,1H),7.56(dd,J=7.1,1.7Hz,1H),7.09(s,1H),6.36(t,J=7.3H z,1H),6.25(s,1H),4.98(td,J=5.7,3.0Hz,0H),4.82(td,J=5.7,3.0Hz,1H),3 .03–2.95(m,1H),2.90(d,J=4.3Hz,3H),1.30-1.18(m,1H),0.96-0.85(m,1H).

[0157] Example 5

[0158] N-cyclopropyl-7-(methylamino)-5-((2-carbonyl-1-(1H-pyrrol-1-yl)-1,2-dihydropyridin-3-yl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxamide (Compound 5)

[0159] Compound 5 was prepared by the following steps:

[0160] Step A: Int-3 (3 g, 13.32 mmol) was dissolved in acetic acid (1 mL) and 2,5-dimethoxytetrahydrofuran (2.29 g, 17.31 mmol) was added. The reaction solution was stirred at 100 ° C for 4 hours. After the reaction was completed, it was cooled and the reaction mixture was concentrated. The residue was added with saturated sodium carbonate solution (150 mL) and extracted with dichloromethane (250 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by silica gel column chromatography (dichloromethane / methanol = 20 / 1) to give compound 5a (3.03 g, yield 82%). LC / MS (ESI + )m / z:276.31[M+H] + .

[0161] Step B: Compound 5a (3.03 g, 11.01 mmol) was dissolved in dichloromethane (9 mL) and trifluoroacetic acid (5 mL) was added. The reaction mixture was stirred at 25°C for 5 hours. LCMS showed that the reaction was complete. The reaction mixture was concentrated to give compound 5b (2.2 g, yield: 73%). LC / MS (ESI + )m / z:176.19[M+H] + .

[0162] Step C: Compound 5b (1.50 g, 8.56 mmol) was dissolved in dioxane (8 mL) and Int-1f (1.52 g, 4.28 mmol), palladium acetate (189 mg, 841 μmol), BINAP (1.07 g, 1.71 mmol) and cesium carbonate (5.58 g, 17.12 mmol) were added respectively. After nitrogen was replaced in the reaction system, the mixture was stirred at 50°C for 2 hours. LC / MS showed that the reaction was complete. The reaction solution was concentrated and the residue was purified by silica gel column chromatography (dichloromethane / methanol = 20 / 1). The crude product was further purified by reverse phase preparative HPLC to obtain compound 5c (879 mg, yield 20%). LC / MS (ESI + )m / z:494.5[M+H] + .

[0163] Step D: Compound 5c (879 mg, 1.78 mmol) was dissolved in methanol (4 mL) and water (3 mL), and sodium hydroxide (399 mg, 10.00 mmol) was added. The reaction solution was stirred at 50°C for 24 hours. LC / MS showed that the reaction was complete. The reaction solution was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 3 / 1). The crude product was further purified by reverse phase preparative HPLC to obtain compound 5d' (420 mg, yield 47%) and compound 5d (120 mg, yield 13%). LC / MS (ESI +)m / z:366.35[M+H] + .

[0164] Step E: Compound 5d (20 mg, 54 μmol) was dissolved in dimethyl sulfoxide (5 mL), and cyclopropaneamine (4.5 mg, 60 μmol), DIPEA (24.7 mg, 191 μmol, 33 μL) and T3P (52 mg, 82 μmol, 50% content) were added respectively. The reaction solution was stirred at room temperature for 12 hours. LC / MS showed that the reaction was complete. The reaction solution was concentrated and the residue was purified by reverse phase preparative HPLC to obtain a white solid, compound 5 (1.2 mg, yield 5%). LC / MS (ESI + )m / z:405.1[M+H] + ; 1 HNMR(400MHz, DMSO-d6)δ9.03(s,1H),8.26–8.19(m,2H),7.94(d,J=4.4Hz,1H),7.81(d,J=4.4Hz,1H),7.48(dd,J=6.8,4.8Hz,1H),7.09( d,J=4.4Hz,2H),6.37(d,J=8.4Hz,1H),6.23–6.18(m,3H),2.89(d,J=4.8Hz,3H),2.89-2.84(m,1H),0.82–0.76(m,2H),0.55–0.50(m,2H).

[0165] Example 6

[0166] N-(3-Hydroxy-2,2-dimethylpropyl)-7-(methylamino)-5-((2-carbonyl-1-(1H-pyrrol-1-yl)-1,2-dihydropyridin-3-yl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxamide (Compound 6)

[0167] Compound 6 was prepared by the following steps:

[0168] Step A: Compound 5d (40 mg, 109 μmol) was dissolved in DMF (7 mL), and 3-amino-2,2-dimethylpropan-1-ol (225 mg, 2.19 mmol), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU, 208 mg, 547 μmol) and N,N-diisopropylethylamine (DIPEA, 283 mg, 2.19 mmol, 381 μL) were added respectively. The reaction solution was stirred at 25 ° C for 4 hours. LC / MS showed that the reaction was complete. The reaction solution was directly purified by reverse phase preparative HPLC to obtain a white solid, compound 6 (6.6 mg, yield 13%). LC / MS (ESI + )m / z:451.2[M+H] + ; 1 HNMR(400MHz,DMSO-d6)δ8.98(s,1H),8.29(dd,J=7.4,4.0Hz,1H),8.23(s,1H) ,7.95(m,J=4.4Hz,1H),7.83(s,1H),7.44(s,1H),7.07(d,J=4.0Hz,2H),6.41(d d,J=7.4Hz,1H),6.24(dd,1H),6.21(d,J=4.4Hz,2H),4.67(dd,J=6.4Hz,1H),3 .27(d,J=6.4Hz,2H), 3.12(d,J=6.4Hz,2H), 2.91(d,J=4.8Hz,3H), 0.84(s,6H).

[0169] Example 7

[0170] N-ethoxy-7-(methylamino)-5-((2-carbonyl-1-(1H-pyrrol-1-yl)-1,2-dihydropyridin-3-yl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxamide (Compound 7)

[0171] Compound 7 was prepared by the following steps:

[0172] Step A: Compound 5d (40 mg, 109.49 μmol) was dissolved in DMSO (5 mL), and O-ethylhydroxylamine hydrochloride (133 mg, 2.19 mmol), N-methylimidazole (NMI, 44 mg, 547 μmol, 43 μL), DIPEA (283 mg, 2.19 mmol, 381 μL) and tetramethylchlorouronium hexafluorophosphate (TCFH, 153 mg, 547 μmol) were added respectively. The reaction solution was stirred at room temperature for 4 hours. LC / MS showed that the reaction was complete. The reaction solution was directly purified by reverse phase preparative HPLC column to obtain a white solid, compound 7 (16.3 mg, yield 36%). LC / MS (ESI + )m / z:409.17[M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ10.37(s,1H),9.03(s,1H),8.49(d,J=8.4Hz,1H),8.24(s,1H),7.92(d,J=4.4Hz,1H),7.44(s,1H),7.08 (t,J=4.4Hz,2H),6.38(s,1H),6.21(dd,J=4.4,4.0Hz,3H),3.96(q,J=8.0Hz,2H),2.90(d,J=4.8Hz,3H),1.25(t,J=8.0Hz,3H).

[0173] Example 8 / Example 9

[0174] N-((1S,2S)-2-Methoxycyclobutyl)-7-(methylamino)-5-((2-carbonyl-1-(1H-pyrrol-1-yl)-1,2-dihydropyridin-3-yl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0175] N-((1R,2R)-2-methoxycyclobutyl)-7-(methylamino)-5-((2-carbonyl-1-(1H-pyrrol-1-yl)-1,2-dihydropyridin-3-yl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0176] Compound 8 and compound 9 were prepared by the following steps:

[0177] Step A: To a dichloromethane solution (5 mL) of compound 5d' (330 mg) were added trans-1,2-aminocyclobutanol (67.9 mg), DIPEA (274 mg) and 1-propylphosphoric anhydride (586 mg, 50% ethyl acetate solution). The reaction mixture was reacted at room temperature for 16 hours. LC / MS showed that the reaction was complete. The reaction system was concentrated and the residue was purified by silica gel column chromatography (dichloromethane / methanol = 99 / 1 to 97 / 3) to obtain a yellow solid, compound 8b (260 mg, yield 68%). LC / MS (ESI + )m / z:535.4[M+H] + .

[0178] Step B: 2,6-di-tert-butyl-4-methylpyridine (46.1 mg) and trimethyloxonium tetrafluoroborate (22.1 mg) were added to a dichloromethane solution (1 mL) of compound 8b (40 mg). The reaction mixture was reacted at room temperature for 16 hours, and LC / MS showed that the reaction was complete. The reaction system was concentrated, and the residue was purified by reverse phase preparative HPLC to obtain a yellow solid 8c (30 mg, yield 73%). LC / MS (ESI + )m / z:549.4[M+H] + .

[0179] Step C: Compound 8c (60 mg) was dissolved in dichloromethane (2 mL). Trifluoroacetic acid (TFA, 0.5 mL) was added and the reaction was stirred at room temperature for 3 hours. LC / MS indicated completion of the reaction. The reaction solution was concentrated and the residue was purified by chiral preparative SFC to afford compound 9 (P1, 14 mg, yellow solid) and compound 8 (P2, 24 mg, yellow solid).

[0180] Compound 8: LC / MS (ESI + )m / z 449.3[M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ9.06 (s, 1H), 8.35 (m, 1H), 8.22 (s, 1H), 8.03 (d, J = 9.1Hz ,1H),7.95(t,J=4.9Hz,1H),7.49(m,1H),7.09(t,J=2.3Hz,2H),6.37(t,J=7.3H z,1H),6.23(s,1H),6.22(t,J=2.3Hz,2H),4.33(m,1H),3.73(q,J=7.6Hz,1H),3 .22(s,3H),2.90(d,J=4.8Hz,3H),2.09(m,2H),1.53(m,1H),1.42(m,1H); Chiral HPLC: retention time = 5.03 min.

[0181] Compound 9: LC / MS (ESI + )m / z:449.3[M+H] + ; 1 H NMR(400MHz, DMSO-d6)δ9.06(s,1H),8.35(dd,J=7.5,1.8Hz,1H),8.21(s,1H),8.03(d,J=9.0Hz,1H),7.95(s,1H),7.49(dd ,J=7.1,1.7Hz,1H),7.09(t,J=2.3Hz,2H),6.37(t,J=7.3Hz,1H),6.23(s,1H),6.22(t,J=2.3Hz,2H),4.33(p,J=8.6Hz,1H), 3.73 (q, J = 7.6 Hz, 1H), 3.22 (s, 3H), 2.90 (s, 3H), 2.10 (m, 2H), 1.58–1.48 (m, 1H), 1.40 (p, J = 9.8 Hz, 1H); Chiral HPLC: retention time = 7.41 min.

[0182] Example 10 / Example 11

[0183] N-((1S,2S)-2-Hydroxycyclobutyl)-7-(methylamino)-5-((2-carbonyl-1-(1H-pyrrol-1-yl)-1,2-dihydropyridin-3-yl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0184] N-((1R,2R)-2-Hydroxycyclobutyl)-7-(methylamino)-5-((2-carbonyl-1-(1H-pyrrol-1-yl)-1,2-dihydropyridin-3-yl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0185] Compounds 10 and 11 were prepared by the following steps:

[0186] Step A: Dissolve 8b (60 mg) in dichloromethane (2 mL) and add TFA (0.5 mL). The reaction mixture was stirred at room temperature for 3 hours. LC / MS indicated completion of the reaction. The reaction mixture was concentrated, and the residue was purified by chiral preparative SFC to afford compound 10 (P1, 8.7 mg, yellow solid) and compound 11 (P2, 25 mg, yellow solid).

[0187] Compound 10: LC / MS (ESI + )m / z:435.3[M+H] + ; 1H NMR (400 MHz, DMSO-d6) δ 9.05 (s, 1H), 8.36 (d, J = 7.4 Hz, 1H), 8.21 (s, 1H), 7.97 (m, 2H), 7.53–7.46 (m, 1H), 7.09 (t, J = 2.4 Hz, 2H), 6.39 (t, J = 7.3 Hz, 1H), 6.24 (s, 1H), 6.22 (m, 2H), 5.43 (d, J = 7.3 Hz, 1H), 4.23 (m, 1H), 3.87 (m, 1H), 2.90 (d, J = 4.7 Hz, 3H), 2.04 (m, 2H), 1.48 (m, 1H), 1.28 (m, 1H); Chiral HPLC: retention time = 2.32 min.

[0188] Compound 11: LC / MS (ESI + )m / z:435.3[M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ9.05 (s, 1H), 8.36 (dd, J = 7.5, 1.7Hz, 1H), 8.21 (s, 1H), 7 .97(m,2H),7.49(dd,J=7.0,1.7Hz,1H),7.09(t,J=2.3Hz,2H),6.39(t,J=7.3Hz, 1H),6.24(s,1H),6.22(m,2H),5.43(d,J=7.1Hz,1H),4.22(m,1H),3.88(m,1H), 2.90(d,J=4.7Hz,3H),2.04(m,2H),1.55–1.44(m,1H),1.30–1.19(m,1H); Chiral HPLC: retention time = 3.45 min.

[0189] Example 12

[0190] N-(2-Fluoroethyl)-7-(methylamino)-5-((2-carbonyl-1-(1H-pyrrol-1-yl)-1,2-dihydropyridin-3-yl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxamide (Compound 12)

[0191] Compound 12 was prepared by the following steps:

[0192] Step A: Compound 5d (40 mg) and 2-fluoroethane-1-amine hydrochloride (70 mg) were dissolved in DMSO (5 mL), and N-methylimidazole (550 mg) and TCFH (220 mg) were added at room temperature. The reaction solution was stirred at 25°C for 16 hours. LC / MS showed that the reaction was complete. The reaction solution was directly purified by preparative HPLC to obtain a white solid, compound 12 (14 mg). LC / MS (ESI + )m / z:411.1[M+H] + ; 1 H NMR(400MHz, DMSO-d6)δ9.01(s,1H),8.36–8.32(m,1H),8.24(s,1H),8.02–7.91(m,2H),7.46–7.43(m,1H),7.11–7.05( m,2H),6.39–6.32(m,1H),6.25(s,1H),6.22–6.20(m,2H),4.68–4.52(m,1H),3.76–3.63(m,2H),2.91(d,J=4.8Hz,3H).

[0193] Example 13

[0194] N-(Cyclopropylmethoxy)-7-(methylamino)-5-((2-carbonyl-1-(1H-pyrrol-1-yl)-1,2-dihydropyridin-3-yl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxamide (Compound 13)

[0195] Compound 13 was prepared by the following steps:

[0196] Step A: Compound 5d (45 mg) and O-(cyclopropylmethyl)hydroxylamine (110 mg) were dissolved in dimethyl sulfoxide (7 mL), and then NMI (50 mg) and TCFH (70 mg) were added at room temperature. The reaction solution was stirred at 25°C for 16 hours. LCMS showed that the reaction was complete. The reaction solution was directly purified by preparative HPLC to obtain a gray solid 13 (3.5 mg). LC / MS (ESI + )m / z:435.1[M+H] + ; 1HNMR(400MHz,DMSO-d6)δ9.01(s,1H),8.36–8.32(m,1H),8.24(s,1H),8.0 2–7.91(m,2H),7.46–7.43(m,1H),7.11–7.05(m,2H),6.39–6.32(m,1H),6 .25(s,1H),6.22–6.20(m,2H),4.68–4.52(m,1H),3.76–3.63(m,2H),2.91 (d,J=4.8Hz,3H),1.15-1.11(m,1H),0.56-0.50(m,1H),0.30-0.27(m,1H).

[0197] Example 14

[0198] 7-(Methylamino)-N-neopentyl-5-((2-carbonyl-1-(1H-pyrrol-1-yl)-1,2-dihydropyridin-3-yl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxamide (Compound 14)

[0199] Compound 14 was prepared by the following steps:

[0200] Step A: Compound 5d (40 mg, 109 μmol) was dissolved in dimethyl sulfoxide, and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (308 mg, 1.10 mmol), N-methylimidazole (90 mg, 1.10 mmol) and 2,2-dimethylpropan-1-amine (95 mg, 1.10 mmol) were added at 0°C. The reaction solution was stirred at room temperature for 30 minutes. LC / MS showed that the reaction was complete. The reaction solution was directly purified by reverse phase preparative HPLC to obtain compound 14 (14.1 mg, yield 29%). LC / MS (ESI + )m / z:435.2[M+H] + ; 1 HNMR(400MHz,DMSO-d6)δ8.98(s,1H),8.28(dd,J=7.6,1.6Hz,1H),8.23(s,1H),7.95(q,J=4.8,4.0Hz,1H),7.77(t,J =6.8Hz,1H),7.45(dd,J=7.0,1.6Hz,1H),7.07(t,J=2.4Hz,2H),6.39(t,J=7.2Hz,1H),6.2 4(s,1H),6.22(t,J=2.4Hz,2H),3.24(d,J=6.8Hz,2H),2.91(d,J=4.8Hz,3H),0.91(s,9H).

[0201] Example 15

[0202] N-((1R,2S)-2-fluorocyclopropyl)-5-((1-(4-hydroxypiperidin-1-yl)-2-carbonyl-1,2-dihydropyridin-3-yl)amino)-7-(methylamino)pyrazolo[1,5-a]pyrimidine-3-carboxamide (Compound 15)

[0203] Compound 15 was prepared by the following steps:

[0204] Step A: 1,4-dioxo-8-azaspiro[4.5]decane 15a (2 g) was dissolved in acetic acid (30 mL) and water (10 mL), and then sodium nitrite (1.16 g) was slowly added dropwise to the reaction solution at 0°C. The reaction was allowed to proceed at room temperature for 1 hour. LC / MS showed that the reaction was complete. The reaction solution was concentrated to dryness and then purified by silica gel column chromatography (petroleum ether / ethyl acetate = 5 / 1) to obtain a yellow solid, compound 15b (2.49 g). LC / MS (ESI + )m / z:173.1[M+H] + .

[0205] Step B: Compound 15b (2.49 g) was dissolved in tetrahydrofuran (25 mL), and then lithium aluminum tetrahydride (551 mg) was added at 0°C. The reaction was allowed to proceed at room temperature for 2 hours. LC / MS showed that the reaction was complete. The reaction solution was slowly quenched with water, filtered, and the filter cake was washed with tetrahydrofuran. The filtrate was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane / methanol = 10 / 1) to obtain a yellow liquid, compound 15c (2 g). LC / MS (ESI + )m / z:159.1[M+H] + .

[0206] Step C: Compound 15c (1.3 g) and triethylamine (1.52 g) were dissolved in ethanol (15 mL), and then compound 1c (1.5 g) was added. The reaction was allowed to proceed at room temperature for 4 hours. LC / MS showed that the reaction was complete. The reaction solution was used directly in the next step without treatment. LC / MS (ESI + )m / z:295.2[M+H] + .

[0207] Step D: Potassium hydroxide (300 mg) was added to the reaction solution obtained in the previous step. The reaction was allowed to proceed at 50°C for 2 hours. LC / MS showed that the reaction was complete. The reaction solution was concentrated to dryness, and the residue was purified by silica gel column chromatography (dichloromethane / methanol = 20:1) to obtain a yellow oil, compound 15e (520 mg). LC / MS (ESI +)m / z:280.9[M+H] + .

[0208] Step E: Compound 15e (500 mg), triethylamine (530 mg) and diphenylphosphoryl azide (1.43 g) were dissolved in tert-butyl alcohol (5 mL). The mixture was reacted at 90°C for 8 hours. LC / MS showed that the reaction was complete. The reaction solution was concentrated to dryness, and the residue was purified by column chromatography (petroleum ether / ethyl acetate = 5:1) to obtain compound 15f (950 mg) as a white solid. LC / MS (ESI + )m / z:296.2[M+H] + .

[0209] Step F: Compound 15f (490 mg, 1.39 mmol) was dissolved in dichloromethane (2 mL), trifluoroacetic acid (2.98 g, 2 mL) was added, and the mixture was stirred at room temperature for 8 hours. LC / MS showed that the reaction was complete. The reaction solution was concentrated, and the residue was purified by reverse phase silica gel column chromatography (acetonitrile / water = 1:20) to obtain compound 15g (50 mg, yield 17%). LC / MS (ESI + )m / z:208.1[M+H] + .

[0210] Step G: Disperse compound 15g (83 mg, 217 μmol), Int-1 (50 mg, 241 μmol), cesium carbonate (117 mg, 361 μmol), BINAP (60 mg, 96 μmol), and palladium acetate (10 mg, 48 μmol) in 1,4-dioxane (2 mL). Stir at 60°C overnight under nitrogen. LC / MS showed the reaction was complete. The reaction solution was concentrated, and the residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 1) to obtain compound 15h (45 mg, yield 33%). LC / MS (ESI + )m / z:555.2[M+H] + .

[0211] Step H: 15h (45 mg, 81 μmol) was dissolved in methanol (1 mL), sodium borohydride (4.6 mg, 121 μmol) was added at 0°C, and the mixture was stirred at room temperature for 1 hour. LC / MS showed that the reaction was complete. After quenching with water, the product was extracted with dichloromethane, and the organic phase was concentrated to give crude 15i, which was used directly in the next step. LC / MS (ESI + )m / z:457.2[M+H] + .

[0212] Step I: The crude product 15i (45 mg) obtained in the previous step was dissolved in dichloromethane (2 mL) and trifluoroacetic acid (2 mL) was added. Stir at room temperature for 1 hour. LC / MS showed that the reaction was complete. The reaction solution was concentrated and the residue was purified by reverse phase preparative HPLC to give compound 15 (17 mg, yield 46%). LC / MS (ESI + )m / z:457.2[M+H] + ; 1 HNMR(400MHz,DMSO-d6)δ8.86(s,1H),8.23(s,1H),8.09(d,J=7.6Hz,1H),7.90( d,J=4.8Hz,1H),7.77(d,J=4.8Hz,1H),7.29(d,J=6.8Hz,1H),6.20(s,1H),6.17( t,J=7.2Hz,1H),4.99–4.76(m,1H),4.69(s,1H),3.59(s,1H),2.98(s,1H),2.90( d,J=4.8Hz,3H),1.83(s,2H),1.59(s,2H),1.28-1.16(m,1H),0.90-0.80(m,1H).

[0213] Example 16

[0214] N-(2-Hydroxyethoxy)-7-(methylamino)-5-((2-carbonyl-1-(1H-pyrrol-1-yl)-1,2-dihydropyridin-3-yl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxamide (Compound 16)

[0215] Compound 16 was prepared by the following steps:

[0216] Step A: Dissolve 2-hydroxyisoindoline-1,3-dione 16a (1 g), 2-[tert-butyl(dimethyl)silyl]oxyethanol 16b (1.08 g), and triphenylphosphine (2.41 g) in anhydrous tetrahydrofuran (20 mL). Replace the mixture with nitrogen three times, stir at 0°C for 10 minutes, and add diisopropyl azodicarboxylate (1.86 g) dropwise. Allow to react at room temperature for 16 hours. TLC indicates the reaction is complete. The reaction solution is concentrated under reduced pressure, and the residue is purified by silica gel column chromatography to afford compound 16c (1.56 g, 79% yield).

[0217] Step B: Dissolve compound 16c (960 mg) and hydrazine hydrate (1 mL) in dichloromethane (5 mL). Reaction was allowed to proceed at room temperature for 2 hours. TLC indicated completion of the reaction. The reaction solution was poured into water (50 mL) and extracted with dichloromethane (20 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to afford compound 16d (300 mg, 52% yield).

[0218] Step C: Dissolve 16d (150 mg), compound 5d (100 mg), DIPEA (138 mg), and HATU (163 mg) in N,N-dimethylformamide (3 mL). Stir at room temperature for 2 hours. LC / MS indicates completion of the reaction. Dilute the reaction solution with 30 mL of water and extract three times with ethyl acetate (10 mL x 3). Combine the organic phases and wash three times with aqueous solution (10 mL x 3). Wash twice with saturated aqueous sodium chloride solution (5 mL x 2). Dry over anhydrous sodium sulfate, filter, and concentrate. The residue is purified by silica gel column chromatography to obtain compound 16e (100 mg, 72% yield).

[0219] Step D: Compound 16e (90 mg) was dissolved in 4 M hydrochloric acid-dioxane solution (2 mL). The reaction solution was stirred at 50°C for 2 hours. LC / MS showed that the reaction was complete. After the reaction solution was concentrated, the residue was purified by reverse phase preparative HPLC to obtain compound 16 (5.0 mg, yield 8%). LC / MS (ESI + )m / z:425.3[M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ10.44(s,1H),9.03(s,1H),8.51(dd,J=7.5,1.7Hz,1 H),8.26(s,1H),8.00–7.91(m,1H),7.46(dd,J=7.1,1.7Hz,1H),7.08(t,J=2 .3Hz,2H),6.43(t,J=7.3Hz,1H),6.21(dd,J=4.2,1.8Hz,3H),4.87(d,J=6.0 Hz, 1H), 3.95 (t, J = 4.9 Hz, 2H), 3.66 (t, J = 5.0 Hz, 2H), 2.90 (d, J = 4.8 Hz, 3H).

[0220] Example 17

[0221] 7-(Methylamino)-5-((2-carbonyl-1-(1H-pyrrol-1-yl)-1,2-dihydropyridin-3-yl)amino)-N-(2,2,2-trifluoroethoxy)pyrazolo[1,5-a]pyrimidine-3-carboxamide (Compound 17)

[0222] Compound 17 was prepared by the following steps:

[0223] Step A: Compound 5d (40 mg) was dissolved in dimethyl sulfoxide (2 mL), and trifluoroethoxyamine hydrochloride (151 mg), DIPEA (70 mg), and HATU (62 mg) were added and stirred at room temperature for 30 minutes. LC / MS showed that the reaction was complete. The reaction solution was directly purified by reverse phase preparative HPLC to obtain compound 17 (11.8 mg, 23% yield). LC / MS (ESI + )m / z:463.2[M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ10.77(s,1H),9.03(s,1H),8.53(d,J=7.4Hz,1H),8.29(s,1H),7.95(d,J=5.0Hz,1H),7.47(dd,J=7.0,1.8Hz ,1H),7.08(t,J=2.4Hz,2H),6.30(t,J=7.2Hz,1H),6.25(s,1H),6.21(t,J=2.4Hz,2H),4.60(q,J=9.0Hz,2H),2.90(d,J=4.6Hz,3H).

[0224] Example 18

[0225] N-isopropoxy-7-(methylamino)-5-((2-carbonyl-1-(1H-pyrrol-1-yl)-1,2-dihydropyridin-3-yl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxamide (Compound 18)

[0226] Compound 18 was prepared by the following steps:

[0227] Step A: Compound 5d (32.8 mg) and HATU (44.3 mg) were dissolved in DMSO (2 mL), and 2-(aminooxy)propane hydrochloride (50 mg) and DIPEA (115.8 mg) were added. The mixture was stirred at room temperature for 30 minutes. LC / MS showed that the reaction was complete. The reaction solution was directly purified by reverse phase preparative HPLC to obtain a white solid, compound 18 (12.3 mg, 32% yield). LC / MS (ESI + )m / z:423.2[M+H] + ; 1H NMR (400MHz, DMSO-d6) δ10.21(s,1H),9.03(s,1H),8.43(dd,J=7.6,1.8Hz,1H),8.24(s,1H),7.94(s,1H),7.46(dd,J=7.2,1.7Hz,1 H),7.08(t,J=2.4Hz,2H),6.33(t,J=7.2Hz,1H),6.23–6.18(m,3H),4.15-4.09(m,1H),2.90(d,J=3.6Hz,3H),1.25(d,J=6.0Hz,6H).

[0228] Example 19

[0229] N-cyclobutyloxy-7-(methylamino)-5-((2-carbonyl-1-(1H-pyrrol-1-yl)-1,2-dihydropyridin-3-yl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxamide (Compound 19)

[0230] Compound 19 was prepared by the following steps:

[0231] Step A: Dissolve N-hydroxyphthalimide (3 g), cyclobutanol (1.33 g), and triphenylphosphine (7.24 g) in anhydrous tetrahydrofuran (50 mL). Replace the mixture with nitrogen three times, stir at 0°C for 10 minutes, and add diisopropyl azodicarboxylate (5.58 g) dropwise. Allow to react at room temperature for 4 hours. TLC indicates completion of the reaction. The reaction mixture is concentrated under reduced pressure, and the residue is purified by silica gel column chromatography to yield Compound 19b (3.9 g, 97% yield) as a white solid. 1 H NMR(400MHz,Chloroform-d)δ7.86(dd,J=5.4,3.2Hz,2H),7.77(dd,J=5.6,3.2Hz,2H) ,4.84–4.76(m,1H),2.43–2.27(m,4H),1.93–1.80(m,1H),1.60(dt,J=8.4,1.2Hz,1H).

[0232] Step B: Dissolve compound 19b (2 g) and hydrazine hydrate (4.61 g) in dichloromethane (20 mL). Allow to react at room temperature for 2 hours. TLC indicates completion of the reaction. Pour the reaction mixture into water (200 mL) and extract with dichloromethane (40 mL x 3). Combine the organic phases, dry over anhydrous sodium sulfate, filter, and concentrate to obtain compound 19c (425 mg). Use directly in the next step.

[0233] Step C: Compound 5d (21 mg), compound 19c (50 mg), HATU (28.4 mg), and DIPEA (22.3 mg) were dissolved in DMSO (3 mL). The mixture was allowed to react at room temperature for 30 minutes. LC / MS showed that the reaction was complete. The reaction solution was directly purified by reverse phase preparative HPLC to obtain a white solid, compound 19 (10.7 mg, 42% yield). LC / MS (ESI + )m / z:435.3[M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ10.27 (s, 1H), 9.03 (s, 1H), 8.45 (dd, J = 7.6, 1.8Hz, 1H), 8.24(s,1H),7.93(d,J=5.2Hz,1H),7.46(dd,J=7.2,1.7Hz,1H),7.08(t,J=2.4H z,2H),6.35(t,J=7.2Hz,1H),6.22–6.20(m,3H),4.48(p,J=7.2Hz,1H),2.90(d, J=4.4Hz,3H),2.24–2.02(m,4H),1.74(q,J=10.4Hz,1H),1.55(p,J=9.6Hz,1H).

[0234] Example 20 / Example 20'

[0235] N-((1R,2R)-2-(Methoxy-d3)-cyclobutyloxy)-7-(methylamino)-5-((2-carbonyl-1-(1H-pyrrol-1-yl)-1,2-dihydropyridin-3-yl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0236] N-((1S,2S)-2-(Methoxy-d3)-cyclobutyloxy)-7-(methylamino)-5-((2-carbonyl-1-(1H-pyrrol-1-yl)-1,2-dihydropyridin-3-yl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0237] Compound 20 and Compound 20' were prepared by the following steps:

[0238] Step A: Trans-1,2-aminocyclobutanol hydrochloride 8a (1.4 g, 16.07 mmol), potassium carbonate (5.55 g, 40.17 mmol), and benzyl bromide (6.05 g, 35.35 mmol, 4.20 mL) were dissolved in DMF (40 mL) and stirred at 50°C for 3 hours. The reaction solution was cooled to room temperature, diluted with ethyl acetate, and washed with saturated NaCl solution. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 0 to 20%) to obtain compound 20a (3.8 g, yield 88%). LC / MS (ESI + )m / z:268[M+H] + .

[0239] Step B: To a solution of compound 20a (130 mg, 486 μmol) in anhydrous tetrahydrofuran was added potassium tert-butoxide (65 mg, 583 μmol) at 0°C. The reaction mixture was stirred for 1 hour, and then deuterated iodomethane (84 mg, 583 μmol) was added. The reaction mixture was stirred for 1 hour at room temperature. The reaction mixture was quenched with water and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 0 to 10%) to give compound 20b (105 mg, yield 75%). LC / MS (ESI + )m / z:285[M+H] + .

[0240] Step C: Compound 20b (1.7 g, 5.98 mmol) and palladium hydroxide on carbon (200 mg) were dispersed in 20 mL of ethanol. After replacing the hydrogen atmosphere, the reaction system was stirred at 40°C under hydrogen balloon pressure for 3 hours. TLC indicated the reaction was complete. The reaction solution was filtered, and a 1 M solution of dioxane hydrochloride (10 mL) was added to the filtrate. The mixture was stirred at room temperature for 1 hour and then concentrated to afford compound 20c (950 mg), which was used directly in the next step.

[0241] Step D: Compound 5d' (100 mg, 214 μmol), HATU (163 mg, 429.68 μmol), and diisopropylethylamine (138 mg, 1.07 mmol) were dissolved in DMF (10 mL), and compound 20c (200 mg) was added. The reaction mixture was stirred at room temperature for 3 hours. LCMS showed that the reaction was complete. Ethyl acetate was added to the reaction solution and diluted, and the solution was washed with saturated ammonium chloride, water, and saturated brine, respectively. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (dichloromethane / methanol = 0 to 10%) to obtain compound 20d (80 mg, yield 67%). LC / MS (ESI + )m / z:552[M+H] + .

[0242] Step E: Compound 20d (100 mg, 181 μmol) was dissolved in dichloromethane (5 mL) and trifluoroacetic acid (2 mL) was added. The reaction was stirred at room temperature for 2 hours. LC / MS indicated completion of the reaction. The reaction solution was concentrated, and the residue was purified by reverse-phase preparative HPLC to afford a mixture of enantiomers (45 mg). This was then resolved by chiral preparative SFC to afford compound 20 (P1, 15 mg) and compound 20' (P2, 15 mg).

[0243] Compound 20: LC / MS (ESI + )m / z:452.3[M+H] + ; 1 H NMR(400MHz,DMSO-d6)δ9.05(s,1H),8.35(d,J=8.4Hz,1H),8.21(s,1H),8.03(d,J =9.2Hz,1H),7.95(d,J=5.2Hz,1H),7.51–7.44(m,1H),7.09(t,J=2.4Hz,2H),6.36 (t,J=7.2Hz,1H),6.22(dd,J=5.2,2.4Hz,3H),4.32(p,J=8.4Hz,1H),3.73(q,J=8. 4,7.2Hz,1H),2.90(d,J=4.8Hz,3H),2.20–2.02(m,2H),1.61–1.34(m,2H); Chiral HPLC: retention time = 4.39 min.

[0244] Compound 20': LC / MS (ESI + )m / z:452.2[M+H] + ; 1 H NMR(400MHz,DMSO-d6)δ9.05(s,1H),8.35(d,J=8.4Hz,1H),8.21(s,1H),8.03(d,J =9.2Hz,1H),7.95(d,J=5.2Hz,1H),7.51–7.44(m,1H),7.09(t,J=2.4Hz,2H),6.36 (t,J=7.2Hz,1H),6.22(dd,J=5.2,2.4Hz,3H),4.32(p,J=8.4Hz,1H),3.73(q,J=8. 4,7.2Hz,1H),2.90(d,J=4.8Hz,3H),2.20–2.02(m,2H),1.61–1.34(m,2H); Chiral HPLC: retention time = 5.60 min.

[0245] Example 21

[0246] N-((1R,2R)-2-ethoxycyclobutyl)-7-(methylamino)-5-((2-carbonyl-1-(1H-pyrrol-1-yl)-1,2-dihydropyridin-3-yl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxamide (Compound 21)

[0247] Compound 21 was prepared by the following steps:

[0248] Step A: Compound 5d (300 mg), trans-1,2-aminocyclobutanol hydrochloride 8a (123 mg), DIPEA (249 mg), and 1-propylphosphonic anhydride (533 mg, 50% ethyl acetate solution) were dissolved in dichloromethane (5 mL). The mixture was stirred at room temperature for 16 hours. LC / MS indicated the reaction was complete. The reaction solution was concentrated, and the residue was purified by silica gel column chromatography (dichloromethane / methanol = 20 / 1) to provide compound 21a (150 mg, 43% yield).

[0249] Step B: Dissolve compound 21a (120 mg), triethyloxonium tetrafluoroborate (127 mg), and 2,6-di-tert-butyl-4-methylpyridine (184 mg) in dichloromethane (10 mL). Stir at room temperature for 16 hours. LC / MS indicated completion of the reaction. The reaction solution was concentrated, and the residue was purified by reverse-phase preparative HPLC to afford compound 21b (100 mg, 79% yield).

[0250] Step C: Compound 21b (90 mg) was dissolved in dichloromethane (3 mL), and trifluoroacetic acid (1 mL) was added. The reaction was stirred at room temperature for 1.5 hours. LC / MS indicated completion of the reaction. The reaction solution was concentrated, and the residue was purified by reverse-phase preparative HPLC and then resolved by chiral preparative SFC to provide compound 21 (P1, 16.4 mg, 22% yield) and compound 21' (P2, 15.8 mg, 21% yield).

[0251] Compound 21: LC / MS (ESI + )m / z:463.3[M+H] + ; 1H NMR(400MHz,DMSO-d6)δ9.05(s,1H),8.41–8.31(m,1H),8.21(s,1H),8.03(d,J=9.3Hz,1H),7 .94(s,1H),7.49(d,J=7.2Hz,1H),7.10(s,2H),6.36(t,J=7.2Hz,1H),6.23(s,3H),4.33(t,J= 11.2 Hz, 1H), 3.80 (d, J = 7.2 Hz, 1H), 3.50 (d, J = 12.0 Hz, 1H), 3.37 (d, J = 7.2 Hz, 1H), 2.90 (s, 3H), 2.15–2.03 (m, 2H), 1.57–1.50 (m, 1H), 1.39 (d, J = 11.2 Hz, 1H), 1.07 (t, J = 7.2 Hz, 3H); Chiral HPLC: retention time = 4.95 min.

[0252] Compound 21': LC / MS (ESI + )m / z:463.3[M+H] + ; 1 H NMR(400MHz,DMSO-d6)δ9.05(s,1H),8.41–8.31(m,1H),8.21(s,1H),8.03(d,J=9.3Hz,1H),7 .94(s,1H),7.49(d,J=7.2Hz,1H),7.10(s,2H),6.36(t,J=7.2Hz,1H),6.23(s,3H),4.33(t,J= 11.2 Hz, 1H), 3.80 (d, J = 7.2 Hz, 1H), 3.50 (d, J = 12.0 Hz, 1H), 3.37 (d, J = 7.2 Hz, 1H), 2.90 (s, 3H), 2.15–2.03 (m, 2H), 1.57–1.50 (m, 1H), 1.39 (d, J = 11.2 Hz, 1H), 1.07 (t, J = 7.2 Hz, 3H); Chiral HPLC: retention time = 8.16 min.

[0253] Example 22 / Example 23

[0254] N-((1S,2S)-2-Hydroxycyclobutyl)-7-(methylamino)-5-((1-morpholino-2-carbonyl-1,2-dihydropyridin-3-yl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0255] N-((1R,2R)-2-Hydroxycyclobutyl)-7-(methylamino)-5-((1-morpholino-2-carbonyl-1,2-dihydropyridin-3-yl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0256] Compound 22 and Compound 23 were prepared by the following steps:

[0257] Step A: Disperse Int-4 (817 mg, 2.31 mmol), compound 3a (900 mg, 4.61 mmol), BINAP (574 mg, 922 μmol), palladium acetate (103 mg, 461 μmol) and cesium carbonate (1.50 g, 4.61 mmol) in dioxane (20 mL). After replacing nitrogen in the reaction system, stir at 60°C overnight. LC / MS showed that the reaction was complete. The reaction solution was concentrated and the residue was purified by reverse phase preparative HPLC to obtain compound 22a (1.2 g, yield 50%). LC / MS (ESI + )m / z:513.5[M+H] + .

[0258] Step B: Compound 22a (1.2 g, 2.34 mmol) and lithium hydroxide (490 mg, 11.68 mmol) were dissolved in a mixture of tetrahydrofuran (10 mL) and water (10 mL) and stirred at 60°C overnight. LC / MS showed that the reaction was complete. The reaction solution was concentrated and the residue was purified by reverse phase preparative HPLC to give compound 22b (300 mg, yield 26%). LC / MS (ESI + )m / z:485.4[M+H] + .

[0259] Step C: Compound 22b (200 mg, 411 μmol), trans-1,2-aminocyclobutanol (53 mg, 617 μmol), 1-propylphosphoric anhydride (196 mg, 50% ethyl acetate solution) and DIPEA (212 mg, 1.65 mmol) were dissolved in N,N-dimethylformamide (5 mL) and stirred at room temperature overnight. LC / MS showed that the reaction was complete. The reaction solution was concentrated and the residue was purified by silica gel column chromatography (dichloromethane / methanol = 10 / 1) to give compound 22c (180 mg, yield 78%). LC / MS (ESI + )m / z:554.6[M+H] + .

[0260] Step D: Compound 22c (50 mg, 90 μmol) was dissolved in dichloromethane (2 mL), trifluoroacetic acid (1 mL) was added, and the mixture was stirred at room temperature for 1 hour. LCMS indicated the reaction was complete. The reaction solution was concentrated, and the residue was purified by reverse-phase preparative HPLC and then by chiral preparative SFC to afford compound 22 (P1, 4.0 mg, 9% yield) and compound 23 (P2, 4.3 mg, 10% yield).

[0261] Compound 22: LC / MS (ESI + )m / z:455.3[M+H] + ; 1 HNMR(400MHz,DMSO-d6)δ8.96(s,1H),8.23(d,J=7.2Hz,1H),8.19(s,1H),8.03(d,J =9.2Hz,1H),7.92(s,1H),7.38–7.34(m,1H),6.25(t,J=7.2Hz,1H),6.22(s,1H),4. 35–4.27 (m, 1H), 3.71–3.65 (m, 2H), 3.19 (s, 3H), 2.90 (s, 3H), 2.13–2.07 (m, 2H), 1.58–1.44 (m, 2H), 1.42–1.33 (m, 2H), 1.31 (d, J = 8.6 Hz, 2H), 0.94–0.83 (m, 2H); Chiral HPLC: retention time = 6.45 min.

[0262] Compound 23: LC / MS (ESI + )m / z:455.3[M+H] + ; 1 HNMR(400MHz,DMSO-d6)δ8.96(s,1H),8.25–8.20(m,1H),8.19(s,1H),8.03(d,J=9.2Hz,1H) ,7.91(d,J=4.0Hz,1H),7.70(d,J=9.2Hz,1H),7.36(d,J=8.4Hz,1H),6.25(t,J=7.2Hz,1H), 6.22 (s, 1H), 4.34–4.28 (m, 1H), 3.71–3.65 (m, 2H), 3.19 (s, 3H), 2.90 (s, 3H), 2.13–2.07 (m, 2H), 1.58–1.43 (m, 2H), 1.40–1.34 (m, 2H), 1.30–1.26 (m, 2H), 0.87 (d, J = 5.6 Hz, 2H); Chiral HPLC: retention time = 9.85 min.

[0263] Example 24 / Example 24'

[0264] N-((1R,2R)-2-(methoxy-d3)cyclobutyl)-7-((methyl-d3)amino)-5-((2-carbonyl-1-(1H-pyrrol-1-yl)-1,2-dihydropyridin-3-yl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0265] N-((1S,2S)-2-(methoxy-d3)cyclobutyl)-7-((methyl-d3)amino)-5-((2-carbonyl-1-(1H-pyrrol-1-yl)-1,2-dihydropyridin-3-yl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0266] Compound 24 and Compound 24' were prepared by the following steps:

[0267] Step A: Disperse 5,7-dichloropyrazolo[1,5-a]pyrimidine-3-carboxylic acid ethyl ester (10.2 g), deuterated methylamine hydrochloride (3.04 g) and potassium carbonate (13.55 g) in a mixed solvent of acetonitrile (100 mL) and water (10 mL) and react at room temperature for 1 hour. LC / MS shows that the reaction is complete. The reaction solution is concentrated under reduced pressure, and water (100 mL) is added to the residue. The mixture is stirred for 2 minutes and then filtered. The filter cake is dried under reduced pressure to obtain a yellow solid, namely compound 24a (4.5 g, yield 44%). LC / MS (ESI + )m / z:258.1[M+H] + .

[0268] Step B: Disperse compound 24a (4 g) and cesium carbonate (7.59 g) in N,N-dimethylformamide (100 mL). Add 4-methoxybenzyl chloride (3.65 g) dropwise at 0°C. After the addition is complete, react at room temperature for 1 hour. LC / MS shows that the reaction is complete. The reaction solution is concentrated under reduced pressure, and the residue is purified by silica gel column chromatography to obtain a yellow-green solid, compound 24b (5.1 g, yield 87%). LC / MS (ESI + ) m / z:378.3[M+H] + .

[0269] Step C: Compound 24b (500 mg), compound 2d (279 mg), BINAP (164 mg), palladium acetate (29 mg) and cesium carbonate (646 mg) were dispersed in dioxane (10 mL). After nitrogen was replaced in the reaction system, it was heated to 60°C for 4 hours. LC / MS showed that the reaction was complete. The reaction solution was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane / methanol = 50 / 1 to 20 / 1) to obtain a green solid, compound 24c (220 mg, yield 32%). LC / MS (ESI + )m / z:517.3[M+H] + .

[0270] Step D: Compound 24c (210 mg) and sodium hydroxide (126 mg) were dissolved in a mixed solution of methanol (5 mL), water (5 mL), and tetrahydrofuran (10 mL). The reaction mixture was heated to 80°C for 8 hours. LC / MS showed that the reaction was complete. The reaction solution was dried under reduced pressure, and the residue was added to water (50 mL). The pH was adjusted to a weak acidic state with 1N hydrochloric acid, and extracted with dichloromethane (30 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a gray solid, compound 24d (120 mg, 60% yield). LC / MS (ESI) m / z: 487.3 [MH] + .

[0271] Step E: Compound 24d (120 mg), compound 20c (255 mg), HATU (186 mg) and DIPEA (95 mg) were dissolved in N,N-dimethylformamide (2 mL). The mixture was allowed to react at room temperature for 30 minutes. LC / MS showed that the reaction was complete. The reaction solution was dried under reduced pressure, and the residue was purified by silica gel column chromatography (dichloromethane / methanol = 50 / 1 to 20 / 1) to obtain a yellow crude product, compound 24e (110 mg). LC / MS (ESI + )m / z:575.5[M+H] + .

[0272] Step F: Compound 24e (100 mg) was dissolved in dichloromethane (9 mL), trifluoroacetic acid (3 mL) was added, and the mixture was stirred at room temperature for 3 hours. LC / MS indicated the reaction was complete. The reaction solution was concentrated, and the residue was purified by reverse-phase preparative HPLC to yield 20 mg of a mixture of enantiomers. This was then resolved by chiral preparative SFC to afford compound 24 (P1, 3.1 mg) and compound 24' (P2, 3.4 mg).

[0273] Compound 24: LC / MS (ESI + )m / z:455.3[M+H] + ; 1H NMR (400MHz, DMSO-d6) δ9.05 (s, 1H), 8.35 (dd, J = 7.6, 1.8Hz, 1H), 8.21 (s, 1H), 8. 03(d,J=9.2Hz,1H),7.92(s,1H),7.49(dd,J=7.2,1.7Hz,1H),7.09(t,J=2.4Hz,2H ),6.37(t,J=7.2Hz,1H),6.22(dd,J=4.0,1.8Hz,3H),4.32(p,J=8.8Hz,1H),3.73 (q,J=7.6Hz,1H),2.20-1.99(m,2H),1.60–1.47(m,1H),1.45-1.30(m,1H); Chiral HPLC: retention time = 7.78 min.

[0274] Compound 24': LC / MS (ESI + )m / z:455.3[M+H] + ; 1 HNMR (400MHz, DMSO-d6) δ9.03 (s, 1H), 8.35 (dd, J = 7.6, 1.8Hz, 1H), 8.21 (s, 1H), 8. 03(d,J=9.2Hz,1H),7.85(s,0H),7.49(dd,J=7.2,1.7Hz,1H),7.09(t,J=2.4Hz,2H) ,6.37(t,J=7.2Hz,1H),6.22(dd,J=4.4,1.9Hz,3H),4.38–4.27(m,1H),3.73(d,J= 7.6Hz,1H),2.16–2.03(m,2H),1.52(q,J=9.6Hz,1H),1.41(q,J=9.6Hz,1H); Chiral HPLC: retention time = 11.48 min.

[0275] Example 25 / Example 25'

[0276] N-((1R,2R)-2-methoxycyclobutyl)-7-(methylamino)-5-((1-morpholino-2-carbonyl-1,2-dihydropyridin-3-yl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0277] N-((1S,2S)-2-methoxycyclobutyl)-7-(methylamino)-5-((1-morpholino-2-carbonyl-1,2-dihydropyridin-3-yl)amino)pyrazolo[1,5-a]pyrimidine-3-carboxamide

[0278] Compound 25 and Compound 25' were prepared by the following steps:

[0279] Step A: Compound 22c (150 mg, 270 μmol), trimethyloxonium tetrafluoroborate (240 mg, 1.62 mmol) and 2,6-di-tert-butyl-4-methylpyridine (222 mg, 1.08 mmol) were dissolved in dichloromethane (5 mL) and stirred at room temperature overnight. LC / MS showed that the reaction was complete. The reaction solution was concentrated and the residue was purified by silica gel column chromatography (dichloromethane / methanol = 20 / 1) to give compound 25a (100 mg, yield 65%). LC / MS (ESI + )m / z:568.6[M+H] + .

[0280] Step B: Compound 25a (50 mg, 90 μmol) was dissolved in dichloromethane (2 mL) and trifluoroacetic acid (1 mL) was added. The mixture was stirred at room temperature for 1 hour. LCMS indicated the reaction was complete. The reaction solution was concentrated, and the residue was purified by reverse-phase preparative HPLC to yield a pair of enantiomers. These were then separated by chiral preparative SFC to afford compound 25 (P1, 4.0 mg, 9% yield) and compound 25′ (P2, 4.3 mg, 10% yield).

[0281] Compound 25: LC / MS (ESI + )m / z:454.4[M+H] + ; 1 HNMR(400MHz, DMSO-d6)δ8.96(s,1H),8.23(dd,J=7.6,1.6Hz,1H),8.18(s,1H),7.98(d,J= 8.8Hz,1H),7.91(q,J=3.6,3.2Hz,1H),7.35(dd,J=7.2,1.6Hz,1H),6.27(t,J=7.2Hz,1H),6 .23(s,1H),5.40(d,J=7.2Hz,1H),4.26–4.15(m,1H),3.88–3.81(m,1H),3.74(s,4H),2.90(d,J=4.8Hz,3H),2.02(d,J=9.6Hz,2H),1.48(t,J=9.2Hz,2H),1.23(t,J=7.2Hz,4H);Chiral HPLC: retention time = 7.83min.

[0282] Compound 25': LC / MS (ESI + )m / z:454.4[M+H] + ; 1HNMR(400MHz,DMSO-d6)δ8.96(s,1H),8.26–8.22(m,1H),8.18(s,1H),7.97(d,J=8.8Hz ,1H),7.91(d,J=4.8Hz,1H),7.35(dd,J=7.1,1.6Hz,1H),6.27(t,J=7.2Hz,1H),6.23(s ,1H),5.40(d,J=7.2Hz,1H),4.21(t,J=8.8Hz,1H),3.87–3.81(m,1H),3.73(s,4H),2.90(d,J=4.8Hz,3H),2.02(d,J=9.6Hz,2H),1.59–1.34(m,2H),1.29–1.18(m,4H);Chiral HPLC: retention time = 9.74min.

[0283] Example 26

[0284] 5-((1-(4,4-difluoropiperidin-1-yl)-2-carbonyl-1,2-dihydropyridin-3-yl)amino)-N-((1R,2S)-2-fluorocyclopropyl)-7-(methylamino)pyrazolo[1,5-a]pyrimidine-3-carboxamide (Compound 26)

[0285] Compound 26 was prepared by the following steps:

[0286] Step A: Compound 15a' (16 mg, 35 μmol) was dissolved in dichloromethane (2 mL) and N-ethyl-N-(trifluorosulfanyl)ethylamine (8.5 mg, 52 μmol) was added at -20°C. The mixture was stirred at 0°C for 2 hours. LC / MS showed that the reaction was complete. The reaction solution was concentrated and the residue was purified by reverse phase preparative HPLC to give compound 26 (1.7 mg, yield 10%). LC / MS (ESI + )m / z:477.3[M+H] + ; 1HNMR(400MHz,DMSO-d6)δ8.91(s,1H),8.23(s,1H),8.12(d,J=7.6Hz,1H),7.9 2(d,J=3.6Hz,1H),7.77(d,J=4.4Hz,1H),7.29(d,J=6.8Hz,1H),6.64(s,1H), 6.22(s,2H),5.32(s,2H),4.99-4.80(m,1H),2.99(s,1H),2.90(d,J=4.4Hz,3 H), 2.15 (s, 2H), 1.99 (s, 2H), 1.47 (d, J = 6.0Hz, 1H), 1.33 (s, 1H), 0.87 (s, 1H).

[0287] TYK2 inhibitor biological screening and results

[0288] Test Example 1: Inhibition test of the compound of the present invention on the TYK2 / JAK1 signaling pathway in PBMC cells

[0289] Human PBMCs (Shanghai Aoneng Biotechnology Co., Ltd., Cat#FPB004F-C) at a density of 500,000 cells were added to each well of a 96-well cell culture plate in a volume of 66.5 μL. 3.5 μL of serially diluted compounds in DMSO were added and mixed thoroughly. The plates were incubated in a 37°C incubator for 60 minutes. IFN-alpha (Biolegend, Cat#592702) was diluted to 600 ng / mL in DPBS containing 0.1% BSA (BI, Cat#02-023-1A). PE-anti-hCD3 antibody (Biolegend, Cat#300308) was diluted 4-fold in DPBS containing 0.1% BSA. After the 60-minute incubation, 6 μL of PE-anti-hCD3 antibody and 4 μL of diluted IFN-alpha were added to each well. The plates were incubated in a 37°C incubator for 30 minutes. Transfer all cells to a 96-well deep-well plate and add 1 mL of 37°C preheated Lyse / Fix Buffer (BD Biosciences, Cat#558049). Incubate at 37°C in the dark for 10 minutes. Centrifuge at 600 g for 5 minutes, discard the supernatant, wash twice with 1 mL of DPBS, and centrifuge again. Add 0.4 mL of Perm Buffer III (BD Biosciences, Cat#558050) to the cell pellet. Incubate at 4°C in the dark for 30 minutes. Centrifuge at 600 g for 5 minutes, discard the supernatant, wash twice with 1 mL of Staining Buffer (0.2% BSA + 1 mM EDTA + DPBS), and centrifuge again. Dilute APC anti-human pSTAT5 antibody (BD Biosciences, Cat#562076) 200-fold in Staining Buffer, add 100 μL to each well, and mix thoroughly. Incubate at room temperature for 40 minutes. Wash twice with 1 mL of Staining buffer per well and centrifuge at 600 g for 5 minutes. Discard the supernatant and resuspend the cell pellet in 200 μL of Staining buffer. Analyze the cells on a Beckman CytoFlex flow cytometer. The inhibitory activity of the compounds of this invention against pSTAT5 in human PBMC cells is shown in Table 1.

[0290] Table 1: Inhibitory activity of the compounds of the present invention on pSTAT5 in human PBMC cells

[0291] The test results show that the compound of the present invention has good inhibitory activity on the Tyk2-STAT5 pathway.

[0292] Test Example 2: Evaluation of the in vitro stability of the compounds of the present invention on human liver microsomal enzymes

[0293] Prepare two 96-well plates, designated T60 and NCF60. Add 445 μL of microsomal working solution (liver microsomal protein concentration 0.56 mg / mL) (Corning, Cat# 452117) to each of the T60 and NCF60 plates. Preincubate the plates in a 37°C water bath for approximately 10 minutes. After preincubation, add 5 μL of the test article or control compound working solution to each of the T60 and NCF60 plates and mix thoroughly. The reaction was initiated by adding 50 μL of potassium phosphate buffer to each well of the NCF60 incubation plate. To the T0 stop plate, 180 μL of stop solution (200 ng / mL tolbutamide and 200 ng / mL labetalol in acetonitrile) and 6 μL of NADPH (nicotinamide adenine dinucleotide phosphate) regeneration system working solution were added. A 54 μL sample was removed from the T60 incubation plate and transferred to the T0 stop plate (T0 sample generation). To the blank plate, only 54 μL of microsome working solution, 6 μL of NADPH regeneration system working solution, and 180 μL of stop solution were added. The reaction was initiated by adding 44 μL of NADPH regeneration system working solution to each well of the T60 incubation plate. Therefore, in the sample of the test or control compound, the final reaction concentration of the compound is 1 μM, the concentration of liver microsomes is 0.5 mg / mL, and the final concentrations of DMSO and acetonitrile in the reaction system are 0.01% (v / v) and 0.99% (v / v), respectively.

[0294] After incubation for the appropriate time (e.g., 5, 15, 30, 45, and 60 minutes), 180 μL of stop solution (containing 200 ng / mL tolbutamide and 200 ng / mL labetalol in acetonitrile) was added to each well of the stop plate. A 60 μL sample was then removed from the T60 or NCF60 incubation plate to terminate the reaction. All sample plates were shaken and centrifuged at 3220 × g for 20 minutes. 80 μL of supernatant was then diluted into 240 μL of pure water for liquid chromatography-tandem mass spectrometry analysis.

[0295] The in vitro elimination rate constants ke of the test and control compounds were calculated by converting the ratio of the compound to the internal standard peak area into the residual percentage using the following formula:

[0296] when

[0297] In vitro liver microsomal intrinsic clearance (CLint (mic) ) and hepatic intrinsic clearance (CLint (liver) )

[0298] CLint (mic) =0.693 / T 1 / 2 / Microsomal protein content (microsomal concentration during incubation mg / mL)

[0299] CLint (liver) =CLint (mic) × amount of microsomal protein in the liver (mg / g) × liver weight to body weight ratio

[0300] The in vitro human liver microsomal enzyme stability data of each compound obtained in Experimental Example 2 are shown in Table 2.

[0301] Table 2: Results of the stability test of the compounds of the present invention on human liver microsomal enzymes

[0302] The test results show that the compound of the present invention has a good clearance rate in human liver microsomes in vitro, showing good pharmacokinetic properties.

Claims

1. A compound as shown in formula I, its stereoisomer or a pharmaceutically acceptable salt thereof, in, R 1 is a C1-C6 alkyl group, 1-1 Substituted C1-C6 alkyl, C1-C6 alkoxy, substituted by one or more R 1-2 Substituted C1-C6 alkoxy, 3-6 membered cycloalkyl, substituted by one or more R 1-3 Substituted 3-6 membered cycloalkyl, 3-7 membered heterocycloalkyl, substituted by one or more R 1-4 Substituted 3-7 membered heterocycloalkyl, -OR 1-5 or one or more R 1-5-1 Substituted-OR 1-5 , the 3-7 membered heterocycloalkyl group and one or more R 1-4 The heteroatom in the 3-7-membered heterocycloalkyl in the substituted 3-7-membered heterocycloalkyl is selected from one or more of N, O and S, and the number of the heteroatoms is 1, 2 or 3; R 1-1 and R 1-2 Each is independently halogen, hydroxyl or 3-6 membered cycloalkyl; R 1-3 Each independently represents halogen, hydroxyl, C1-C6 alkyl, or one or more R 1-3-1 Substituted C1-C6 alkyl, C1-C6 alkoxy or one or more R 1-3-2 Substituted C1-C6 alkoxy; R 1-3-1 and R 1-3-2 each independently is deuterium or a halogen; R 1-4 Each independently represents halogen, hydroxyl, C1-C6 alkyl, or one or more R 1-4-1 Substituted C1-C6 alkyl, C1-C6 alkoxy or one or more R 1-4-2 Substituted C1-C6 alkoxy; R 1-4-1 and R 1-4-2 each independently is deuterium or a halogen; R 1-5 is a 3-6 membered cycloalkyl group; R 1-5-1 Each is independently halogen, hydroxyl or C1-C6 alkyl; Ring A is a 4-12 membered heterocyclic ring or a 5-10 membered heteroaromatic ring, wherein the heteroatoms in the 4-12 membered heterocyclic ring and the 5-10 membered heteroaromatic ring include at least one nitrogen atom, and the remaining heteroatoms are selected from one or more of N, O and S, and the number of heteroatoms is 1, 2 or 3; R 2 Each is independently halogen, hydroxyl, cyano, oxo (=O), C1-C6 alkyl, 2-1 Substituted C1-C6 alkyl, C1-C6 alkoxy or one or more R 2-2 Substituted C1-C6 alkoxy; R 2-1 and R 2-2 are each independently halogen or hydroxy; R 3 is a C1-C6 alkyl group, 1-1 Substituted C1-C6 alkyl, C1-C6 alkoxy, substituted by one or more R 1-2 Substituted C1-C6 alkoxy, 3-6 membered cycloalkyl, substituted by one or more R 1-3 Substituted 3-6 membered cycloalkyl, 3-7 membered heterocycloalkyl, substituted by one or more R 1-4 Substituted 3-7 membered heterocycloalkyl, -OR 1-5 or one or more R 1-5-1 Substituted-OR 1-5 , the 3-7 membered heterocycloalkyl group and one or more R 1-4 The heteroatom in the 3-7-membered heterocycloalkyl in the substituted 3-7-membered heterocycloalkyl is selected from one or more of N, O and S, and the number of the heteroatoms is 1, 2 or 3; R 4 is hydrogen, halogen, hydroxyl, cyano, oxo, C1-C6 alkyl, 1-1 Substituted C1-C6 alkyl, C1-C6 alkoxy, substituted by one or more R 1-2 Substituted C1-C6 alkoxy, 3-6 membered cycloalkyl, substituted by one or more R 1-3 Substituted 3-6 membered cycloalkyl, 3-7 membered heterocycloalkyl, substituted by one or more R 1-4 Substituted 3-7 membered heterocycloalkyl, -OR 1-5 or one or more R 1-5-1 Substituted-OR 1-5 , the 3-7 membered heterocycloalkyl group and one or more R 1-4 The heteroatom in the 3-7-membered heterocycloalkyl in the substituted 3-7-membered heterocycloalkyl is selected from one or more of N, O and S, and the number of the heteroatoms is 1, 2 or 3; m is 0, 1, 2 or 3.

2. The compound of formula I according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, characterized in that: The compound as shown in formula I satisfies one or more of the following conditions: (1) each C1-C6 alkyl group is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentane, isopentane or neopentane, for example methyl, ethyl, isobutyl or neopentane; (2) each C1-C6 alkoxy group is independently a C1-C4 alkoxy group, preferably a methoxy group, an ethoxy group, an n-propoxy group, an isopropoxy group, an n-butoxy group, an isobutoxy group, a sec-butoxy group or a tert-butoxy group, for example, a methoxy group, an ethoxy group or an isopropoxy group; (3) each 3-6 membered cycloalkyl group is independently a 3-6 membered monocyclic cycloalkyl group, preferably cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, such as cyclopropyl or cyclobutyl; (4) each 3-7 membered heterocycloalkyl group is independently a 3-7 membered monocyclic heterocycloalkyl group, preferably a 3-7 membered monocyclic heterocycloalkyl group having one or two heteroatoms selected from N and O; (5) each halogen is independently fluorine, chlorine, bromine or iodine, for example fluorine or chlorine; (6) Each 4-12 membered heterocycle is a 4-12 membered monocyclic heterocycle or a 4-12 membered polycyclic heterocycle; the 4-12 membered monocyclic heterocycle may be a 4-6 membered monocyclic heterocycle, preferably a 4-6 membered monocyclic heterocycle wherein the heteroatom is selected from one or two of N and O, for example The 4-12 membered polycyclic heterocycle is preferably (7) The heteroatom in each 5-10 membered heteroaromatic ring is N, and the number of heteroatoms is 1 or 2; and (8) each 5-10 membered heteroaromatic ring is a 5-6 membered heteroaromatic ring, preferably a 5-6 membered heteroaromatic ring having 1 or 2 heteroatoms selected from N, for example 3. The compound of formula I according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, characterized in that: The compound as shown in formula I satisfies one or more of the following conditions: (1)R 1 is a C1-C6 alkyl group, 1-1 Substituted C1-C6 alkyl, C1-C6 alkoxy, substituted by one or more R 1-2 Substituted C1-C6 alkoxy, 3-6 membered cycloalkyl, substituted by one or more R 1-3 Substituted 3-6 membered cycloalkyl or -OR 1-5 Preferably, R 1 is a C1-C6 alkyl group, 1-1 Substituted C1-C6 alkyl, C1-C6 alkoxy, substituted by one or more R 1-2 Substituted C1-C6 alkoxy, one or more R 1-3 Substituted 3-6 membered cycloalkyl or -OR 1-5 More preferably, R 1 is a C1-C6 alkoxy group, 1-2 Substituted C1-C6 alkoxy or -OR 1-5 ; (2)R 1-1 are each independently halogen or hydroxy; (3)R 1-2 Each is independently fluorine, hydroxyl or 3-6 membered cycloalkyl; (4)R 1-3 Each independently represents halogen, hydroxyl, C1-C6 alkyl, C1-C4 alkoxy or is replaced by one or more R 1-3-2 Substituted C1-C4 alkoxy; preferably halogen, hydroxy, C1-C4 alkoxy or one or more R 1-3-2 Substituted C1-C4 alkoxy; (5)R 1-3-2 are each independently deuterium or fluorine, preferably deuterium; (6) Ring A is a 4-6 membered monocyclic heterocyclic ring or a 5-6 membered heteroaromatic ring, wherein the heteroatoms in the 4-6 membered monocyclic heterocyclic ring and the 5-6 membered heteroaromatic ring are selected from 1 or 2 of N and O; preferably, Ring A is a 6 membered monocyclic heterocyclic ring or a 5 membered heteroaromatic ring, wherein the heteroatoms in the 6 membered monocyclic heterocyclic ring are selected from 1 or 2 of N and O, and the heteroatoms in the 5 membered heteroaromatic ring are selected from N, and the number of heteroatoms is 1 or 2; (7)R 2 Each independently represents halogen, hydroxyl, C1-C6 alkyl, or one or more R 2-1 Substituted C1-C6 alkyl, or C1-C6 alkoxy; preferably halogen or hydroxy; (8)R 2-1 are each independently a halogen; (9)R 3 is a C1-C6 alkyl group; (10)R 4 is hydrogen; and (11)m is 0, 1 or 2.

4. The compound of formula I, its stereoisomer or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, characterized in that: The compound as shown in formula I satisfies one or more of the following conditions: (1)R 1 for (2)R 2 is hydroxy, fluorine, chlorine, methyl, methoxy or trifluoromethyl; (3) Ring A is Preferably, for More preferably, for 5. The compound of formula I according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, characterized in that: The compound as shown in formula I is any of the following compounds:

6. A pharmaceutical composition, comprising: (1) substance A, wherein substance A is a compound of formula I as described in any one of claims 1 to 5, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, and (2) Pharmaceutically acceptable excipients.

7. Use of the compound of formula I according to any one of claims 1 to 5, its stereoisomer or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 6 in the preparation of a TYK2 inhibitor.

8. Use of the compound of formula I as described in any one of claims 1 to 5, its stereoisomer or pharmaceutically acceptable salt thereof, or the pharmaceutical composition as described in claim 6 in the preparation of a medicament for treating a disease mediated by TYK2; the disease may be an autoimmune disease, an inflammatory disease, a bone disease, a metabolic disease, a neurodegenerative disease, cancer, a cardiovascular disease, an allergy, asthma and Alzheimer's disease.

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