Irak degraders and uses thereof

Abstract

The application provides a compound shown in formula I and a use thereof in preparation of a medicine. The application provides a new compound with IRAK4 degradation effect, which can effectively degrade IRAK4 or inhibit the activity of IRAK4 in other ways. The compound has very good application prospects in IRAK4-mediated diseases, including immune diseases, tumors, Alzheimer's disease and fibrosis diseases, and the like, and provides a new selection for screening and / or preparing a medicine for a disease related to the activity of IRAK4 in the clinic.

Description

Technical Field

[0001] This invention relates to a class of compounds with IRAK degradation activity and their use in the preparation of pharmaceuticals. Background Technology

[0002] Protein degradation is a highly regulated and essential process for maintaining cellular homeostasis. The ubiquitin-proteasome pathway (UPP) in vivo plays a crucial role in the selective identification and removal of excess proteins and the degradation of misfolded or abnormal proteins. Ubiquitin molecules are covalently linked to terminal lysine residues via E3 ubiquitin ligases, thereby labeling proteins before they are degraded into small peptides by the proteasome and ultimately digested into their constituent amino acids. These amino acids then serve as building blocks for new proteins. UPP plays a central role in many cellular processes, and its defects or imbalances can contribute to the pathogenesis of various diseases. UPP is central to the regulation of almost all cellular processes, including antigen processing, apoptosis, organelle biogeneration, cell cycle, DNA transcription and repair, differentiation and development, immune responses and inflammation, neural and muscular degeneration, neural network morphogenesis, regulation of cell surface receptors, ion channels and secretory pathways, responses to stress and extracellular regulators, ribosome biogeneration, and viral infection. Defective proteasome degradation is associated with a variety of clinical conditions, including Alzheimer's disease, Parkinson's disease, Huntington's disease, muscular dystrophy, cardiovascular disease, and cancer.

[0003] Protein degradation-targeting chimeras (PROTACs) are an effective method for degrading pathogenic proteins. They involve linking small molecules that can bind to the target protein with E3 ligases, including CRBN, VHL, MDM2, and DRAF, to form heterobifunctional molecules. These molecules then ubiquitinate the target protein via a ubiquitin-proteasome pathway (UPP), thereby enabling proteasome degradation. Compared to small molecule inhibitors, a potential advantage of protein degradation-targeting chimeras is their ability to remove all functions of the pathogenic protein.

[0004] Currently, over 600 E3 ubiquitin ligases have been identified that promote the in vivo ubiquitination of various proteins. These can be classified into four families: the HECT domain E3 family, the U-box E3 family, the monomeric RING E3 family, and the multi-subunit E3 family. Cereblon (CRBN) ligase is the most widely used E3 ligase in PROTAC technology. Cereblon belongs to the Cullin RING E3 ubiquitin ligase family and is a 442-amino acid protein that forms the Cullin-4-RING E3 ubiquitin ligase (CRL4) complex and interacts with the adapter protein-damaged DNA-binding protein 1 (DDB1). In the CRL4 complex, CRBN acts as a substrate-specific receptor. Known CRBN ligands include thalidomide and other derived immunomodulatory imide drugs. Upon ligand binding, the E3 ubiquitin ligase activity of CRBN is reregulated, leading to increased recruitment of transcription factors Ikaros and Aiolos, thereby triggering subsequent ubiquitination and proteasome degradation. Currently, CRBN, as an E3 ligase in PROTACs, has been successfully used to target more than 30 different proteins, including proteins associated with various cancers (SunX et al., 2019), proteins related to immune dysfunction (Bassi et al., 2018), proteins related to neurodegenerative diseases (Silva et al., 2019), and hepatitis C virus proteins (de Wispelaere et al., 2019). Most CRBN-targeting PROTACs employ pomalidoxamine, 4-hydroxythalidomide, alkyl-linked thalidomide derivatives, or lenalidomide derivatives. However, it is possible to develop superior CRBN ligands. These novel CRBN ligands will provide more options for the development of PROTAC technology.

[0005] IRAK4 is a serine / threonine kinase and a key protein mediating interleukin-1 (IL-1) receptor family (IL-1, IL-18, and IL-33 receptors) and pathogen recognition of Toll-like receptor (TLR) signals. Studies have shown that upon recognition of foreign pathogens and inflammatory stress, interleukin-1 receptors or TLR receptors, under the action of extracellular ligands, recruit the adaptor protein Mydoid Differentiation Primary Reactive Protein (Myd88), which then forms a complex with IRAK4, activating the NF-κB light chain enhancer and activator protein 1 (AP-1). This leads to the production of various inflammatory factors, such as tumor necrosis factor-α (TNFα) and IL-6, thereby inducing various immune diseases, such as psoriasis, hidradenitis suppurativa, atopic dermatitis, rheumatoid arthritis, and systemic lupus erythematosus. Furthermore, IRAK4 has been shown to be associated with lymphocytic leukemia and lymphoma, Alzheimer's disease, and fibrotic diseases. Therefore, IRAK4 is a highly attractive target for drug development.

[0006] Currently, major pharmaceutical companies are advancing IRAK4 small molecule inhibitors into clinical trials for the treatment of hematologic malignancies, psoriasis, rheumatoid arthritis, enteritis, and systemic lupus erythematosus. Among them, Pfizer's IRAK4 inhibitor PF-06650833 has entered Phase II clinical trials. Early clinical results indicate that PF-06650833 has a good safety profile, and its efficacy shows that it can inhibit the IRAK4-mediated inflammatory pathway. These clinical data fully demonstrate that IRAK4 is a clinically validated drug target with the potential to treat a variety of diseases.

[0007] Recent studies have shown that, in addition to the kinase activity-mediated inflammatory signaling pathways, the IRAK4 protein cytoskeleton can also activate certain inflammatory signaling pathways. In human skin fibroblasts, ATP-competitive small-molecule inhibition of IRAK4 is ineffective in suppressing the release of IL-6 and TNF-α stimulated by IL-1β. Furthermore, knockout of IRAK4 effectively eliminates IL-1, IL-8, and TLR ligand-mediated inflammatory responses. Therefore, ATP-competitive small-molecule inhibitors cannot completely eliminate the inflammatory signaling pathways mediated by the IRAK4 protein. This demonstrates the therapeutic limitations of targeting IRAK4 with small-molecule inhibitors.

[0008] Protein degradation-targeting chimeras (PROTACs) are an effective means of degrading pathogenic proteins. They involve linking small molecules that bind to the target protein with E3 ligases, including CRBN, VHL, MDM2, and DRAF, to form heterobifunctional molecules. These molecules then ubiquitinate the target protein via a ubiquitin-proteasome pathway (UPP), enabling proteasome degradation. Compared to small molecule inhibitors, a potential advantage of PROTACs is their ability to remove all functions of the pathogenic protein, resulting in broader and deeper inhibition of inflammatory factors and potentially overcoming the limited clinical efficacy of small molecules. Furthermore, GSK scientists have demonstrated that combining a small IRAK4 inhibitor with ligands of E3 ligases CRBN and VHL via a linker fragment to form PROTACs can achieve IRAK4 protein degradation. Kymera and Avinas have also designed corresponding PROTACs targeting IRAK4. These emerging technologies provide a novel therapeutic approach for targeting IRAK4. Summary of the Invention

[0009] This invention provides a compound of Formula I, or a stereoisomer thereof, or a deuterated compound thereof, or a pharmaceutically acceptable salt thereof:

[0010]

[0011] in,

[0012] R 1 Selected from -NR 1a R 1b 4-10 cyclic heterocyclic groups, 5-12 bridging heterocyclic groups, or 5-12 spirocyclic heterocyclic groups, wherein the heterocyclic group, bridging heterocyclic group, or spirocyclic group may be optionally divided by one, two, or three R 11 replace;

[0013] R 1a R 1b Selected independently from hydrogen and C 1～6 Alkyl, C 2～6 alkenyl, C 2～6 Alkyne- and halogen-substituted C 1～6 Alkyl, halogen-substituted C 2～6 Alkenyl, halogen-substituted C 2～6 alkynyl group, -C 0～2 Alkyl-3 to 10-membered carbocyclic groups or -C 0～2 Alkylene-4 to 10-membered heterocyclic groups;

[0014] Each R 11 Each element is independently selected from hydrogen, halogen, and C. 1～6 Alkyl, C2～6 alkenyl, C 2～6 Alkyne- and halogen-substituted C 1～6 Alkyl, halogen-substituted C 2～6 Alkenyl, halogen-substituted C 2～6 alkynyl group, -C 0～2 Alkylene-OR 12 -C 0～2 Alkylene-NR 12 R 13 -C 0～2 Alkylene-C(O)R 12 -C 0～2 Alkylene-NR 12 C(O)R 13 -C 0～2 Alkylene-C(O)NR 12 R 13 -C 0～2 Alkyl-3 to 10-membered carbocyclic groups or -C 0～2 Alkylene-4 to 10-membered heterocyclic groups;

[0015] R 12 R 13 Selected independently from hydrogen and C 1～6 Alkyl, C 2～6 alkenyl, C 2～6 Alkyne- and halogen-substituted C 1～6 Alkyl, halogen-substituted C 2～6 Alkenyl, halogen-substituted C 2～6 alkynyl group, -C 0～2 Alkyl-3 to 10-membered carbocyclic groups or -C 0～2 Alkylene-4 to 10-membered heterocyclic groups;

[0016] n1 and n2 are independently selected from 0, 1 or 2 respectively;

[0017] U is selected from N or CR U ;

[0018] R U Selected from hydrogen or C 1～6 alkyl;

[0019] V is selected from chemical bonds, O, S, or CR. V1 R V2 ;

[0020] R V1 R V2 Each is independently selected from hydrogen or C 1～6 alkyl;

[0021] Ring A is selected from 5-membered aromatic heterocycles and 6-membered aromatic heterocycles, wherein the aromatic heterocycle may be optionally surrounded by one, two or three Rs. A replace;

[0022] Each R A Each element is independently selected from hydrogen, halogen, cyano, and C. 1～6 Alkyl, C 2～6 alkenyl, C 2～6 Alkyne- and halogen-substituted C 1～6 Alkyl, halogen-substituted C 2～6 Alkenyl or halogen-substituted C 2～6 alkynyl group;

[0023] Indicates a single bond or a double bond;

[0024] X is selected from CR X or NR X ;

[0025] Y is selected from N or C(O);

[0026] Z is selected from N or C;

[0027] Q is selected from N or CR Q ;

[0028] R X Selected from hydrogen, halogen, cyano, C 1～6 Alkyl, C 2～6 alkenyl, C 2～6 Alkyne- and halogen-substituted C 1～6 Alkyl, halogen-substituted C 2～6 Alkenyl, halogen-substituted C 2～6 alkynyl group, -C 0～2 Alkylene-OR X1 -C 0～2 Alkylene-NR X1 R X2 -C 0～2 Alkyl-3 to 10-membered carbocyclic groups or -C 0～2 Alkylene-4 to 10-membered heterocyclic groups;

[0029] R X1 R X2 Selected independently from hydrogen and C 1～6 Alkyl, C 2～6 alkenyl, C 2～6 Alkyne- and halogen-substituted C 1～6 Alkyl, halogen-substituted C 2～6 Alkenyl or halogen-substituted C 2～6 alkynyl group;

[0030] R Q Selected from hydrogen, halogens, C 1～6 Alkyl, C 2～6 alkenyl, C 2～6 Alkyne- and halogen-substituted C 1～6Alkyl, halogen-substituted C 2～6 Alkenyl or halogen-substituted C 2～6 Alkyne group.

[0031] In some preferred embodiments, when ring A is selected from a 5-membered aromatic heterocycle, Y is selected from C(O).

[0032] V is selected from chemical bonds, which means that the U and A rings are directly connected by chemical bonds.

[0033] The term "single bond" or "double bond" means that a person skilled in the art can freely choose a chemically feasible single bond or double bond based on X, Y, and Z.

[0034] Furthermore,

[0035] R 1 Selected from -NR 1a R 1b 5-membered nitrogen-containing heterocyclic group, 6-membered nitrogen-containing heterocyclic group, 7-membered nitrogen-containing heterocyclic group, 8-membered nitrogen-containing heterocyclic group, 6-membered nitrogen-containing bridged heterocyclic group, 7-membered nitrogen-containing bridged heterocyclic group, 8-membered nitrogen-containing bridged heterocyclic group, 9-membered nitrogen-containing bridged heterocyclic group, 10-membered nitrogen-containing bridged heterocyclic group, 6-membered nitrogen-containing spiroheterocyclic group, 7-membered nitrogen-containing spiroheterocyclic group, 8-membered nitrogen-containing spiroheterocyclic group, 9-membered nitrogen-containing spiroheterocyclic group, or 10-membered nitrogen-containing spiroheterocyclic group, wherein the heterocyclic group, bridged heterocyclic group, and spiroheterocyclic group may be optionally composed of one, two, or three R... 11 replace.

[0036] Furthermore,

[0037] R 1 Selected from

[0038] Furthermore,

[0039] n1 is selected from 0, n2 is selected from 1; or n1 is selected from 1, n2 is selected from 0; or n1 is selected from 1, n2 is selected from 1; or n1 is selected from 2, n2 is selected from 1; or n1 is selected from 1, n2 is selected from 2;

[0040] U is selected from N or CH;

[0041] V is selected from chemical bonds, O, or CH2;

[0042] Q is selected from N or CH.

[0043] In some embodiments of the present invention, the compound represented by Formula I has the structure represented by Formula IIa or Formula IIb:

[0044]

[0045] in,

[0046] Ring A is selected from a 6-membered aromatic heterocycle, wherein the aromatic heterocycle may optionally be composed of one, two, or three R groups. A replace;

[0047] Each R A Each element is independently selected from hydrogen, halogen, cyano, and C. 1～6 Alkyl, C 2～6 alkenyl, C 2～6 Alkyne- and halogen-substituted C 1～6 Alkyl, halogen-substituted C 2～6 Alkenyl or halogen-substituted C 2～6 alkynyl group;

[0048] R X Selected from hydrogen, halogen, cyano, C 1～6 Alkyl, C 2～6 alkenyl, C 2～6 Alkyne- and halogen-substituted C 1～6 Alkyl, halogen-substituted C 2～6 Alkenyl, halogen-substituted C 2～6 alkynyl group, -C 0～2 Alkylene-OR X1 -C 0～2 Alkylene-NR X1 R X2 -C 0～2 Alkyl-3 to 10-membered carbocyclic groups or -C 0～2 Alkylene-4 to 10-membered heterocyclic groups;

[0049] R X1 R X2 Selected independently from hydrogen and C 1～6 Alkyl, C 2～6 alkenyl, C 2～6 Alkyne- and halogen-substituted C 1～6 Alkyl, halogen-substituted C 2～6 Alkenyl or halogen-substituted C 2～6 Alkyne group.

[0050] In some embodiments of the present invention, further,

[0051] Ring A is selected from

[0052] R X Selected from hydrogen or C 1～6 Alkyl group. Preferably, R X Selected from methyl.

[0053] In some embodiments of the present invention, the compound represented by Formula I has the structure represented by Formula III:

[0054]

[0055] in,

[0056] Ring A is selected from 5-membered aromatic heterocycles and 6-membered aromatic heterocycles, wherein the aromatic heterocycle may be optionally surrounded by one, two or three Rs. A replace;

[0057] Each R A Each element is independently selected from hydrogen, halogen, cyano, and C. 1～6 Alkyl, C 2～6 alkenyl, C 2～6 Alkyne- and halogen-substituted C 1～6 Alkyl, halogen-substituted C 2～6 Alkenyl or halogen-substituted C 2～6 alkynyl group;

[0058] R X Selected from hydrogen, halogen, cyano, C 1～6 Alkyl, C 2～6 alkenyl, C 2～6 Alkyne- and halogen-substituted C 1～6 Alkyl, halogen-substituted C 2～6 Alkenyl, halogen-substituted C 2～6 alkynyl group, -C 0～2 Alkylene-OR X1 -C 0～2 Alkylene-NR X1 R X2 -C 0～2 Alkyl-3 to 10-membered carbocyclic groups or -C 0～2 Alkylene-4 to 10-membered heterocyclic groups;

[0059] R X1 R X2 Selected independently from hydrogen and C 1～6 Alkyl, C 2～6 alkenyl, C 2～6 Alkyne- and halogen-substituted C 1～6 Alkyl, halogen-substituted C 2～6 Alkenyl or halogen-substituted C 2～6 Alkyne group.

[0060] In some embodiments of the present invention, further,

[0061] Ring A is selected from

[0062] R X Selected from hydrogen or C 1～6 Alkyl group. Preferably, R X Selected from methyl.

[0063] In some specific embodiments of the present invention, the compound specifically includes:

[0064]

[0065]

[0066]

[0067] This invention also provides the use of any of the above-mentioned compounds, or their stereoisomers, or their deuterated compounds, or their pharmaceutically acceptable salts, in the preparation of medicaments for the treatment and prevention of one or more diseases related to or mediated by the interleukin-1 receptor-associated kinase 4 (IRAK4) signaling pathway, the interleukin-6 (IL-6) receptor, and tumor necrosis factor α (TNFα). The inventors of this invention have discovered that the compounds of this invention, as IRAK4 degrading agents, can effectively degrade IRAK4, or effectively inhibit downstream molecules of the pathway such as the interleukin-6 (IL-6) receptor and tumor necrosis factor α (TNFα), thereby inhibiting IRAK4.

[0068] Furthermore, the diseases mentioned include cancer, neurodegenerative diseases, viral diseases, autoimmune diseases, inflammatory diseases, genetic diseases, hormone-related diseases, metabolic disorders, organ transplant-related diseases, immunodeficiency diseases, bone-destructive diseases, proliferative diseases, infectious diseases, thrombin-induced platelet aggregation, liver diseases, lesions caused by T-cell activation, and cardiovascular diseases.

[0069] The present invention also provides a pharmaceutical composition comprising any of the above-mentioned compounds, or their stereoisomers, or their deuterated compounds, or their pharmaceutically acceptable salts, and a pharmaceutically acceptable excipient prepared thereof.

[0070] The beneficial effects of this invention include at least the following: the compounds disclosed in this invention that have IRAK4-degrading activity can effectively degrade IRAK4 or otherwise inhibit its activity. They show great promise for application in IRAK4-mediated diseases, including immune diseases, tumors, Alzheimer's disease, and fibrotic diseases, providing a new option for clinical screening and / or preparation of drugs for diseases related to IRAK4 activity.

[0071] The compounds and derivatives provided in this invention can be named according to the IUPAC (International Union of Pure and Applied Chemistry) or CAS (Chemical Abstracts Service, CoLumbus, OH) nomenclature system.

[0072] Regarding the definition of terms used in this invention: Unless otherwise stated, the initial definitions provided for groups or terms herein apply to the groups or terms used throughout this specification; for terms not specifically defined herein, the meanings that a person skilled in the art would give them should be given based on the disclosure and context.

[0073] "Substitution" refers to the replacement of hydrogen atoms in a molecule by other different atoms or molecules.

[0074] "Optionally replaceable" means that "replacement" may but does not have to occur, and this statement includes situations in which it may or may not occur.

[0075] The minimum and maximum carbon atom content in hydrocarbon groups are indicated by a prefix, for example, the prefix C. a～b Alkyl indicates any alkyl group containing one to two carbon atoms ("a" to "b"). Therefore, for example, "C..." 1～4 "Alkyl" refers to an alkyl group containing 1 to 4 carbon atoms.

[0076] In this invention, "alkyl" refers to a saturated hydrocarbon chain having a specified number of member atoms. For example, C 1～6 Alkyl refers to an alkyl group having 1 to 6 member atoms, for example 1 to 4 member atoms. Alkyl groups can be straight-chain or branched. Representative branched alkyl groups have one, two, or three branches. Alkyl groups may optionally be substituted by one or more substituents as defined herein. Alkyl groups include methyl, ethyl, propyl (n-propyl and isopropyl), butyl (n-butyl, isobutyl, and tert-butyl), pentyl (n-pentyl, isopentyl, and neopentyl), and hexyl. Alkyl groups may also be part of other groups, such as C 1～6 Alkyl group.

[0077] In this invention, "alkylene" refers to a divalent saturated aliphatic hydrocarbon group having a specified number of carbon atoms. "C" a～b "Alkylene" refers to an alkylene group having a to b carbon atoms. Alkylene groups include branched and straight-chain hydrocarbon groups. For example, "C 1～6 The term "alkylene" is intended to include methylene, ethylene, propylene, 2-methylpropylene, dimethylethylene, pentylene, etc. Therefore, the term "propylene" can be exemplified by the following structures: Similarly, the term "dimethylbutylene" can be exemplified, for example, by any of the following structures: For example, "C0 alkylene" means that there is a chemical bond here, directly connecting the two parts through a chemical bond.

[0078] In this invention, "alkenyl" refers to a straight-chain or branched hydrocarbon group having a specified number of carbon atoms and at least one vinyl unsaturated site (>C=C<). For example, C a-b Alkenyl refers to an alkenyl group having a to b carbon atoms and is intended to include, for example, vinyl, propenyl, isopropenyl, 1,3-butadienyl, etc.

[0079] In this invention, "alkynyl" refers to a straight-chain monovalent hydrocarbon group or a branched monovalent hydrocarbon group containing at least one triple bond. The term "alkynyl" is also intended to include those hydrocarbon groups having one triple bond and one double bond. For example, C 2-6 The term "alkynyl" is intended to include ethynyl, propynyl, etc.

[0080] In this invention, "chemical bond" refers to a direct connection at that point via a single chemical bond;

[0081] The "halogen" mentioned in this invention is fluorine, chlorine, bromine or iodine.

[0082] In this invention, "halogenated alkyl" and "halogen-substituted alkyl" refer to alkyl groups in which the hydrogen atom can be replaced by one or more halogen atoms. For example, C 1～4 Halogenated alkyl groups refer to alkyl groups containing 1 to 4 carbon atoms in which one or more hydrogen atoms are replaced by one or more halogen atoms. Examples include trifluoromethyl and difluoromethyl.

[0083] The substituents such as "=O" and "=S" mentioned in this invention refer to oxygen or sulfur atoms replacing two hydrogen atoms to form a double bond, or replacing lone pairs of electrons to form a double bond.

[0084] In this invention, "-OR", "-NRR", etc., refer to the R group being connected to an oxygen atom or a nitrogen atom by a single bond.

[0085] In this invention, the oxygen atom in "-C(O)R", "-S(O)2R", etc., is connected to a carbon atom, sulfur atom, or phosphorus atom by a double bond, and R is connected to a carbon atom or sulfur atom by a single bond. In this invention, the oxygen atom in "-C(O)NRR", "-S(O)2NRR", etc., is connected to a carbon atom or sulfur atom by a double bond, the nitrogen atom is connected to a carbon atom or sulfur atom by a single bond, and R is connected to a nitrogen atom by a single bond. In this invention, one R in "-NRC(O)R", "-NRS(O)2R", etc., is connected to a nitrogen atom by a single bond, the other R is connected to a carbon atom or sulfur atom by a single bond, the nitrogen atom is connected to a carbon atom or sulfur atom by a single bond, and the oxygen atom is connected to a carbon atom or sulfur atom by a double bond.

[0086] In this invention, "carbocyclic" and "carbocyclic group" refer to saturated or partially saturated cyclic groups having multiple carbon atoms and no heterocyclic atoms, and having a single ring or multiple rings (fused). The carbon atoms include their oxidation state, such as C(O). For polycyclic systems having aromatic and non-aromatic rings without heterocyclic atoms, the terms "carbocyclic" and "carbocyclic group" (e.g., 5,6,7,8,-tetrahydronaphthalene-5-yl) are used when the junction is located on a non-aromatic carbon atom. The terms "carbocyclic" and "carbocyclic group" include cycloalkenyl groups, such as cyclohexenyl. Examples of carbocyclic groups include, for example, cyclopropyl, cyclobutyl, cyclohexyl, cyclopentyl, cyclooctyl, cyclopentenyl, and cyclohexenyl. Examples of carbocyclic groups including multi-dicarbonyl ring systems are dicyclohexyl, dicyclopentyl, dicyclooctyl, etc. Two such dicarbonyl polycyclic structures are illustrated and named below: Dicyclohexyl and Dicyclohexyl.

[0087] In this invention, "bridged ring" refers to a saturated or partially saturated cyclic group formed by multiple ring bridges having multiple carbon atoms and no cyclic heteroatoms. The term "bridged ring" also includes adamantane systems with adamantane alkyl groups, including but not limited to the following structures.

[0088] In this invention, "heterocyclic" and "heterocyclic group" refer to a saturated ring or a non-aromatic unsaturated ring containing at least one heteroatom and having a single ring or multiple rings (fused); where heteroatoms refer to nitrogen atoms, oxygen atoms, sulfur atoms, etc. Carbon atoms and heteroatoms include their oxidation states, such as C(O), S(O), S(O)2, etc. The terms "heterocyclic" and "heterocyclic group" also apply to polycyclic systems having aromatic and non-aromatic rings containing cyclic heteroatoms, for example... This typically represents a saturated or partially unsaturated monocyclic or bicyclic ring system with multiple ring atoms. Examples of monocyclic saturated heterocyclic groups are oxo-heterobutyl, azirrobutyl, pyrrolyl, 2-oxo-pyrrolyl-3-yl, tetrahydrofuranyl, tetrahydro-thiophenyl, pyrazolyl, imidazoyl, thiazoyl, piperidinyl, tetrahydropyranyl, tetrahydrothiaranyl, piperazinyl, morpholinyl, thiomorpholinyl, 1,1-dioxo-thiomorpholin-4-yl, azirrohepyl, diazirrohepyl, homopiperazinyl, or oxaazirrohepyl. Examples of bicyclic saturated heterocyclic groups are 8-aza-bicyclo[3.2.1]octyl, quinine cycloyl, 8-oxa-3-aza-bicyclo[3.2.1]octyl, and 9-aza-bicyclo[3.3.1]nonyl. Examples of partially unsaturated heterocyclic groups are dihydrofuranyl, imidazolinyl, tetrahydropyridyl, or dihydropyranyl.

[0089] The “bridged heterocycle” mentioned in this invention refers to a saturated or partially saturated cyclic group formed by multiple ring bridges containing at least one heteroatom.

[0090] In this invention, "aromatic ring" and "aryl" refer to an aromatic hydrocarbon group having multiple carbon atoms. Aryl groups typically comprise monocyclic, bicyclic, or tricyclic aryl groups. Furthermore, the term "aryl" as used herein refers to an aromatic substituent that can be a single aromatic ring or multiple aromatic rings fused together. Non-limiting examples include phenyl, naphthyl, or tetrahydronaphthyl.

[0091] In this invention, "aromatic heterocycle" and "aromatic heterocyclic group" refer to an aromatic unsaturated ring containing at least one heteroatom; wherein the heteroatom refers to a nitrogen atom, oxygen atom, sulfur atom, etc. Typically, it refers to an aromatic monocyclic or bicyclic hydrocarbon containing multiple ring atoms, one or more of which are selected from O, N, and S heteroatoms. Preferably, it has one to three heteroatoms. Examples of heterocyclic aryl groups include: pyridyl, indolyl, quinoxalinyl, quinolinyl, isoquinolinyl, benzothiopheneyl, benzofuranyl, benzothiopheneyl, benzopyranyl, benzothiapyranyl, furanyl, pyrroleyl, thiazolyl, oxazolyl, isoxazolyl, triazolyl, tetrazolyl, pyrazolyl, imidazoleyl, thiopheneyl, oxadiazolyl, benzimidazoleyl, benzothiazolyl, and benzoxazolyl.

[0092] The term "pharmaceutically acceptable" means that a carrier, delivery substance, diluent, excipient, and / or the salt formed therefrom is generally chemically or physically compatible with other components constituting a drug dosage form and physiologically compatible with receptors.

[0093] The terms "salt" and "pharmaceutical salt" refer to acidic and / or basic salts formed by the above-described compounds or their stereoisomers with inorganic and / or organic acids and bases, including zwitterionic salts (internal salts) and quaternary ammonium salts, such as alkylammonium salts. These salts can be obtained directly during the final separation and purification of the compounds. Alternatively, they can be obtained by mixing the above-described compounds or their stereoisomers with an appropriate amount (e.g., equimolar amounts) of an acid or base. These salts may be obtained by precipitating in solution and collecting by filtration, by recovery after solvent evaporation, or by freeze-drying after reaction in an aqueous medium. The salts described in this invention can be hydrochlorides, sulfates, citrates, benzenesulfonates, hydrobromides, hydrofluoric acids, phosphates, acetates, propionates, succinates, oxalates, malates, succinates, fumarates, maleates, tartrates, or trifluoroacetates of the compounds.

[0094] The terms "compound of the present invention" or "active ingredient of the present invention" are used interchangeably to refer to stereoisomers, enantiomers, or pharmaceutical salts of the general formula compound. The term also includes racemic mixtures, optical isomers, isotopic compounds (such as deuterated compounds), or leads.

[0095] "Stereoisomers" are compounds composed of identical atoms bonded by the same bonds, but with different three-dimensional structures. This invention will cover various stereoisomers and mixtures thereof.

[0096] When the compounds of the present invention contain alkene double bonds, unless otherwise stated, the compounds of the present invention are intended to contain both E- and Z- geometric isomers.

[0097] "Tautomer" refers to an isomer formed when a proton is transferred from one atom of a molecule to another atom of the same molecule. All tautomer forms of the compounds of this invention are also included within the scope of this invention.

[0098] The compounds of the present invention, or pharmaceutically acceptable salts thereof, may contain one or more chiral carbon atoms, and thus may produce enantiomers, diastereomers, and other stereoisomers. Each chiral carbon atom may be defined as (R)- or (S)- based on stereochemistry. The present invention aims to include all possible isomers, as well as their racemic and optically pure forms. The preparation of the compounds of the present invention may select racemic, diastereomer, or enantiomer as starting materials or intermediates. Optically active isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques, such as crystallization and chiral chromatography.

[0099] Conventional techniques for preparing / separating individual isomers include chiral synthesis from suitable optically pure precursors, or resolution of racemates (or racemates of salts or derivatives) using, for example, chiral high-performance liquid chromatography.

[0100] This invention also includes isotopically labeled compounds, equivalent to the original compounds disclosed herein. However, it is common practice for one or more atoms to be substituted with atoms of different atomic weights or mass numbers. Examples of isotopes of compounds that can be included in this invention include hydrogen, carbon, nitrogen, oxygen, fluorine, and chlorine isotopes. Compounds of this invention, or enantiomers, diastereomers, isomers, or pharmaceutical salts or solvates, containing isotopic or other isotopic atoms of the aforementioned compounds, are all within the scope of this invention. Certain isotopically labeled compounds of this invention, such as radioactive isotopes, are also present and are useful in tissue distribution experiments of drugs and substrates. For example, tritium, i.e., 3H, and carbon-14, i.e., 14C, are relatively easy to prepare and detect and are preferred isotopes. Furthermore, heavier isotope substitutions such as deuterium, i.e., 2H, may be preferred in certain cases due to their excellent metabolic stability, which is advantageous in some therapies, such as increasing half-life or reducing dosage in vivo. Isotope-labeled compounds can be prepared using conventional methods, by replacing non-isotopic reagents with readily available isotope-labeling reagents, according to the schemes disclosed in the examples. In this application, the term "pharmaceutical salt" includes pharmaceutical acid addition salts.

[0101] The compounds involved in this application and their pharmaceutically acceptable salt metabolites, as well as prodrugs that can be converted in vivo into structures of the compounds involved in this application and their pharmaceutically acceptable salts, are also included in the claims of this application.

[0102] Pharmaceutical Compositions and Administration

[0103] The pharmaceutical compositions of this invention are intended for the prevention and / or treatment of one or more diseases associated with or mediated by the interleukin-1 receptor-associated kinase 4 (IRAK4) signaling pathway, the interleukin-6 (IL-6) receptor, and tumor necrosis factor α (TNFα). In this application, "pharmaceutical composition" refers to a formulation of the compound of this invention with a medium generally accepted in the art for delivering a bioactive compound to a mammal (e.g., a human). This medium includes pharmaceutically acceptable carriers. The purpose of the pharmaceutical composition is to facilitate administration to an organism, thereby facilitating the absorption of the active ingredient and the exertion of its bioactivity. The term "pharmaceutical" as used herein refers to a substance (such as a carrier or diluent) that does not affect the bioactivity or properties of the compound of this invention and is relatively non-toxic, i.e., that the substance can be administered to an individual without causing an adverse biological response or adversely interacting with any component contained in the composition.

[0104] In this application, "pharmaceutical excipients" include, but are not limited to, any adjuvants, carriers, excipients, flow aids, sweeteners, diluents, preservatives, dyes / coloring agents, flavoring agents, surfactants, wetting agents, dispersants, suspending agents, stabilizers, isotonic agents, solvents, or emulsifiers that are permitted by the relevant government regulatory authorities to be acceptable for human or livestock use.

[0105] The term “prevention” as used in this article includes reducing the likelihood of a patient developing or worsening a disease or condition.

[0106] The term "treatment" and other similar synonyms used in this article include the following meanings:

[0107] (i) To prevent the occurrence of disease or condition in mammals, especially when such mammals are susceptible to the disease or condition but have not yet been diagnosed with it;

[0108] (ii) To suppress a disease or symptom, that is, to curb its development;

[0109] (iii) To alleviate a disease or symptom, that is, to cause the condition of the disease or symptom to subside; or

[0110] (iv) To alleviate the symptoms caused by the disease or condition.

[0111] As used herein, the terms "effective amount," "therapeutic effective amount," or "pharmaceutical effective amount" refer to an amount of at least one drug or compound that, when taken, is sufficient to alleviate, to some extent, one or more symptoms of the disease or condition being treated. The result may be a reduction and / or relief of signs, symptoms, or causes, or any other desired change in a biological system. For example, an "effective amount" for treatment is the amount of a composition containing the compounds disclosed herein that is clinically necessary to provide significant symptom relief. Effective amounts suitable for any individual case can be determined using techniques such as dose escalation testing.

[0112] In some embodiments, one or more compounds of the present invention may be used in combination with each other. Alternatively, the compounds of the present invention may be used in combination with any other active agent to prepare a medicament or pharmaceutical composition for regulating cell function or treating disease. If a group of compounds is used, these compounds may be administered to the test subject simultaneously, separately, or sequentially.

[0113] Obviously, based on the above description of the present invention, and according to common technical knowledge and conventional methods in the field, various other modifications, substitutions or alterations can be made without departing from the basic technical concept of the present invention. Detailed Implementation

[0114] The present invention is further illustrated below by way of examples, but the invention is not limited to the scope of the examples described. Experimental methods in the following examples, unless otherwise specified, were performed according to conventional methods and conditions, or as selected in the product instructions.

[0115] The raw materials and equipment used in the specific embodiments of the present invention are all known products, obtained by purchasing commercially available products.

[0116] Unless otherwise specified in the examples, the reaction temperature is room temperature, which is defined as 20–25°C. All temperatures are expressed in °C (degrees Celsius).

[0117] The overnight stay is 14±1h.

[0118] High-performance liquid chromatography (HPLC) determination conditions: Waters high-performance liquid chromatograph (e2695 / e2487). Analytical HPLC conditions: C18 column (3.5 μm, 4.6 x 75 mm), UV detection bands of 220 and 254 nm, elution conditions: gradient wash with 5-95% acetonitrile (containing 0.1% V / V TFA or 10 mmol NH4CO3) for 10 min.

[0119] Reversed-phase purification was performed using the GiLson GX-281 reversed-phase preparative chromatograph or the Biotage IsoLera One rapid purification system.

[0120] NMR measurements were performed using a Bruker Avance III 400 or 600 NMR spectrometer. NMR shifts (δ) were expressed in terms of 10⁻⁶. -6 The unit (ppm) is given. Solvents include deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCL3), and deuterated methanol (CD3OD), etc., and the internal standard is tetramethylsilane (TMS).

[0121] The known starting materials, reagents, and solvents of this invention can be synthesized using or according to methods known in the art, or can be purchased from companies such as Chengdu Jinshan Chemical Reagent Co., Ltd., Shanghai Bid Pharmaceutical Technology Co., Ltd., and Shanghai Titan Technology Co., Ltd.

[0122] The known starting materials, reagents, and solvents of this invention can be synthesized using or according to methods known in the art, or can be purchased from companies such as Chengdu Jinshan Chemical Reagent Co., Ltd., Shanghai Bid Pharmaceutical Technology Co., Ltd., and Shanghai Titan Technology Co., Ltd.

[0123] In the above discussion and the following examples, the following abbreviations have the following meanings. If an abbreviation is not defined, it has the generally accepted meaning. MPLC is medium-pressure preparative chromatography; TLC is thin-layer chromatography; MeOH is methanol; EtOH is ethanol; DMAP refers to 4-dimethylaminopyridine; DMF is N,N-dimethylformamide; DMA is N,N-dimethylacetamide; EA is ethyl acetate; THF is tetrahydrofuran; DMSO is dimethyl sulfoxide; DCM is dichloromethane; DCE is dichloroethane; MTBE is methyl tert-butyl ether; Boc2O is ditert-tert-butyl dicarbonate; Boc is tert-butyloxycarbonyl; SEMCl is 2-(trimethylsilyl)ethoxymethyl chloride; SEM is 2-(trimethylsilyl)ethoxymethyl; CbzCl is benzyloxyformyl chloride; Cbz is benzyloxyformyl; FmocCl is 9-fluorenyl chloroformate; Fmoc 9-fluorenylmethoxyformyl; MsCl is methanesulfonyl chloride; Ms is methanesulfonyl; TBSCl is tert-butyldimethylchlorosilane; TBS is tert-butyldimethylchlorosilyl; TBDPSCl is tert-butyldiphenylchlorosilane; TBDPS is tert-butyldiphenylsilyl; TBAF is tetrabutylammonium fluoride; NBS is N-bromosuccinimide; TFA is trifluoroacetic acid; DBU is 1,8-diazabicycloundec-7-ene; DIPEA is N,N-diisopropylethylamine; TEA is triethylamine; HATU is 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate; HBTU is 2-(benzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate.

[0124] Example

[0125] The following are examples of intermediate preparation:

[0126] Synthesis of Intermediate 1: tert-butyl (3-(difluoromethyl)-1-((trans)-4-formylcyclohexyl)-1H-pyrazole-4-yl)carbamate

[0127]

[0128] Step 1: Synthesis of intermediate 1b

[0129] Compound 1a (850 g, 3.47 mmol) was added to methanol, followed by 1 / 200 of concentrated sulfuric acid, and refluxed at 65 °C for 12 h. After the reaction was complete, a saturated sodium carbonate solution was added to adjust the pH to 7, and the mixture was filtered and concentrated. 2000 mL of EA was added, and the mixture was extracted three times with saturated brine. The extract was dried over 400 mL x 3 anhydrous sodium sulfate solutions and concentrated to obtain compound 1b (900 g), which was directly added to the next step.

[0130] Step 2: Synthesis of intermediate 1c

[0131] Compound 1b (900g) was added to THF (6000mL), and lithium aluminum hydride 150g was added in small amounts several times at -20℃. After the reaction was complete, an equal amount of water and an equal amount of 15% sodium hydroxide aqueous solution were added sequentially, followed by quenching with three times the volume of aqueous solution. The mixture was then filtered and concentrated to obtain compound 1c (410g).

[0132] Step 3: Synthesis of intermediate 1d

[0133] Compound 1c (400g) was added to DCM (6000mL) and compound 4 (251g) in an ice-water bath. A DCM solution of 521g TBSCl was added dropwise. The reaction was carried out at room temperature for 12 hours. After the reaction was complete, the mixture was concentrated, washed three times with water (2000mL), washed twice with DCM, and the organic phase was extracted once with 1000mL of saturated brine. The mixture was dried over anhydrous sodium sulfate and concentrated to obtain compound 1d (737g).

[0134] Step 4: Synthesis of intermediate 1e

[0135] Compound 1d (720 g, 2.95 mol) was dissolved in DCM (10 L), and a solution of 940 g of TsCl in dichloromethane (2 eq) was rapidly added dropwise, followed by a slow addition of DMAP in DCM solution. The reaction was carried out at 30 °C for 12 h. After the reaction was complete, 2 L of water was added, and the mixture was extracted twice with 2 L of DCM. The organic phase was extracted once with 10% citric acid, washed once with saturated NaHCO3, and once with saturated brine. The mixture was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography to obtain compound 6 (760 g).

[0136] Step 5: Synthesis of intermediate 1f

[0137] 3-(difluoromethyl)-4-nitro-1H-pyrazole (73.64 g) was added to DMF (1 L) and cesium carbonate (1.4 eq), and reacted for 10 min. Compound 1e (180 g) was added, and the reaction was carried out at 60 °C for 12 h. 2500 mL of water was added, and the mixture was extracted three times with 500 mL of petroleum ether. The mixture was washed twice with 500 mL of water and once with 500 mL of saturated brine. The mixture was dried over anhydrous sodium sulfate, concentrated, and passed through a chromatography column to obtain compound 1f (52.6 g).

[0138] Step Six: Synthesis of 1g of intermediate

[0139] Compound 8 (52.6 g) was mixed with 500 mL of methanol, 1 eq of triethylamine, and 3 eq of (Boc)₂O. The mixture was stirred at room temperature for 10 min, then 0.05 eq of palladium on carbon was added. The mixture was purged with hydrogen three times and allowed to react overnight at room temperature. After the reaction was complete, the mixture was filtered and concentrated to obtain compound 9 (62 g).

[0140] Step 7: Synthesis of intermediate for 1 hour

[0141] 1 g (62 g) of the compound was added to 300 mL of THF. Under an ice-water bath, HF.py (2 eq) was added dropwise, and the mixture was allowed to return to room temperature naturally and stirred for 12 h. After the reaction was complete, the mixture was extracted three times with water and EA, washed once with saturated sodium bicarbonate, washed once with saturated brine, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain the compound (39.6 g). ESI-LCMS: m / z 346 [M+1], 1.45 min.

[0142] Step 8: Synthesis of tert-butyl(3-(difluoromethyl)-1-((trans)-4-formylcyclohexyl)-1H-pyrazole-4-yl)carbamate 1

[0143] Compound 1 (39.6 g) was added to 250 mL of DCM. Under ice-water bath conditions, DMP (1.3 eq) was added in small, repeated additions, and the mixture was stirred for 3 h. After the reaction was complete, the pH was adjusted to 7 with saturated sodium bicarbonate. The mixture was filtered, separated, dried over anhydrous sodium sulfate, concentrated, and purified by chromatography to obtain compound 1 (31 g). ESI-LCMS: m / z 344 [M+1], 1.55 min.

[0144] Synthesis of Intermediate 2: 5-morpholinylpyrazolo[1,5-a]pyrimidine-3-carboxylic acid

[0145]

[0146] Step 1: Synthesis of intermediate 2b

[0147] Ethyl 5-chloropyrazolo[1,5-a]pyrimidine-3-carboxylate (2.25 g, 10 mmol) was dissolved in 10 mL of acetonitrile, followed by the addition of morpholine (1.31 g, 15 mmol) and DIPEA (2.58 g, 20 mmol), and the mixture was heated under reflux for 2 hours. After the reaction was complete, 10 mL of water was added, and a solid precipitated. The solid was cooled, filtered, and the filter cake was washed with water. The solid was collected, dried, and yielded 2.55 g of ethyl 5-morpholinylpyrazolo[1,5-a]pyrimidine-3-carboxylate. LCMS (ESI) m / z: [M+1] = 277.

[0148] Step 2: Synthesis of Intermediate 2

[0149] Ethyl 5-morpholinylpyrazolo[1,5-a]pyrimidine-3-carboxylic acid (2.55 g, 9.2 mmol) was dissolved in 20 mL of methanol, and then a solution of sodium hydroxide (800 mg) in water (10 mL) was added. The mixture was stirred at room temperature for 16 hours. After the reaction was complete, 1 M dilute hydrochloric acid was added to adjust the pH to approximately 5-6, and a solid precipitated. The organic solvent was removed under reduced pressure, and the mixture was then filtered and washed with water. The solid was collected and dried to give 2.01 g of 5-morpholinylpyrazolo[1,5-a]pyrimidine-3-carboxylic acid. LCMS (ESI) m / z: [M+1] = 249.

[0150] Synthesis of intermediate 3: 5-((1R,4R)-2-oxa-5-azabicyclo[2.2.1]heptane-5-yl)pyrazol[1,5-a]pyrimidine-3-carboxylic acid

[0151]

[0152] The preparation method of intermediate 3 is similar to that of intermediate 2.

[0153] Synthesis of intermediate 4: 5-(1,4-oxazaheptan-4-yl)pyrazol[1,5-a]pyrimidine-3-carboxylic acid

[0154]

[0155] The preparation method of intermediate 4 is similar to that of intermediate 2.

[0156] Synthesis of intermediates 5 to 7: The preparation methods of intermediates 5 to 7 are similar to those of intermediate 2, and their numbering and structure are shown in Table 1.

[0157] Table 1: Numbering and Structure of Intermediates 5 to 7

[0158]

[0159] Synthesis of intermediate 8: 3-(4-(6-(4-(((1r,4r)-4-(4-amino-3-(difluoromethyl)-1H-pyrazol-1-yl)cyclohexyl)methyl)piperazin-1-yl)pyridin-3-yl)-3-methyl-1H-indazol-1-yl)piperidine-2,6-dione

[0160]

[0161] Step 1: Synthesis of intermediate 8c

[0162] Compound 8a (600 mg) and compound 8b (870 mg) were dissolved in 6 mL of a mixed solution of dioxane and water (10 / 1, V / V). Under nitrogen protection, K₂CO₃ (515 mg) and Pd(dppf)Cl₂ (136 mg) were added successively. The mixture was evacuated, nitrogen was switched three times, and the mixture was heated to 95 °C and reacted for 4 hours. The reaction solution was filtered, concentrated, and dispersed in 5 mL of acetonitrile. 387 mg of CDI was added, and the mixture was heated to reflux. After the reaction was completed, the solution was concentrated and column chromatography was performed to obtain 8c 400 mg. LCMS (ESI) m / z: [M+1] = 505.24.

[0163] Step 2: Synthesis of intermediate 8d

[0164] Compound 8c (400 mg, 0.794 mmol) was added to dichloromethane (8 mL), followed by HCl / dioxane (8 mL), and the reaction was carried out at room temperature for 1 hour. The mixture was monitored by LCMS, and the crude compound 5d was concentrated to obtain 400 mg. The LCMS (ESI) m / z: [M+1] = 404.46.

[0165] Step 3: Synthesis of compound 8e

[0166] Compound 8d (400 mg, 0.990 mmol), compound 1 (309 mg, 0.900 mmol), and molecular sieve (400 mg) were added to a DMF / THF (1 / 5) (4 mL). Triethylamine (273 mg, 2.701 mmol) was added, and the mixture was stirred for 0.5 h in an ice-salt bath at -10 °C. Acetic acid (162 mg, 2.701 mmol) and sodium borohydride acetate (573 mg, 2.701 mmol) were added, and the mixture was reacted at -10 °C for 3 h under LC-MS monitoring. The reaction was quenched with water, filtered, and the filtrate was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and subjected to silica gel column chromatography (DCM:MeOH = 10:1) to give compound 8e (544 mg, yield 75.17%). LC-MS (ESI) m / z: [M+1] = 731.83

[0167] Step 4: Synthesis of Compound 8

[0168] Compound 8e (544 mg, 0.743 mmol) was added to dichloromethane (6 mL), followed by HCl / dioxane (6 mL), and stirred at room temperature for 1 h. The mixture was then concentrated directly by LCMS to obtain crude compound 8 with LCMS (ESI) m / z: [M+1] = 631.72.

[0169] Synthesis of intermediate 9: 3-(4-(6-(4-(((1r,4r)-4-(4-amino-3-(difluoromethyl)-1H-pyrazol-1-yl)cyclohexyl)methyl)piperazin-1-yl)pyrimidin-5-yl)-3-methyl-1H-indazol-1-yl)piperidin-2,6-dione

[0170]

[0171] The synthesis and preparation methods for intermediate 9 are the same as those for intermediate 8, steps one to four. LCMS(ESI) m / z: [M+1] = 632.71.

[0172] Synthesis of Intermediate 10: 3-(4-(6-(4-(((1r,4r)-4-(4-amino-3-(difluoromethyl)-1H-pyrazol-1-yl)cyclohexyl)methyl)piperazin-1-yl)pyridin-3-yl)-3-methyl-2-oxa-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidine-2,6-dione

[0173]

[0174] The synthesis and preparation methods for intermediate 10, steps one to four, are the same as those for intermediate 8. LCMS(ESI) m / z: [M+1] = 648.2.

[0175] Synthesis of intermediate 11: 3-(4-(1-(1-(((1r,4r)-4-(4-amino-3-(difluoromethyl)-1H-pyrazol-1-yl)cyclohexyl)methyl)piperidin-4-yl)-1H-pyrazol-4-yl)-3-methyl-2-oxa-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidin-2,6-dione

[0176]

[0177] The synthesis and preparation methods for intermediate 11, steps one to four, are the same as those for intermediate 8. LCMS(ESI) m / z: [M+1] = 636.2.

[0178] Synthesis of intermediate 12: 3-(4-((1-(1-(((1r,4r)-4-(4-amino-3-(difluoromethyl)-1H-pyrazol-1-yl)cyclohexyl)methyl)piperidin-4-yl)methyl)-1H-pyrazol-4-yl)-3-methyl-2-oxa-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidin-2,6-dione

[0179]

[0180] The synthesis and preparation methods for intermediate 12 are the same as those for intermediate 8, steps one to four. LCMS(ESI) m / z: [M+1] = 650.3.

[0181] Synthesis of intermediate 13: 3-(4-(2-((1-(1-(((1r,4r)-4-(4-amino-3-(difluoromethyl)-1H-pyrazol-1-yl)cyclohexyl)methyl)piperidin-4-yl)oxy)pyrimidin-5-yl)-3-methyl-1H-indazol-1-yl)piperidin-2,6-dione

[0182]

[0183] Step 1: Synthesis of Compound 13b

[0184] Compound 13a (2 g), pinacol diboronate (2.13 g), and potassium acetate (1.1 g) were added to dioxane (20 mL), followed by the addition of Pd(dppf)Cl2 (0.204 g). The reaction was carried out overnight at 85 °C under nitrogen protection. The mixture was monitored by LCMS, filtered, and the filtrate was concentrated. The concentrate was then obtained by silica gel column chromatography (PE:EA = 1:1) to give compound 13b (900 mg). LCMS (ESI) m / z: [M+1] = 405.13.

[0185] The synthesis and preparation method of intermediate 13, steps two to five, is the same as the synthesis and preparation method of intermediate 8, steps one to four. LCMS(ESI) m / z: [M+1] = 647.72.

[0186] Synthesis of intermediate 14: 3-(4-(6-((1-(1-((((1r,4r)-4-(4-amino-3-(difluoromethyl)-1H-pyrazol-1-yl)cyclohexyl)methyl)piperidin-4-yl)oxy)pyridin-3-yl)-3-methyl-1H-indazol-1-yl)piperidin-2,6-dione

[0187]

[0188] Step 1: Synthesis of compound 14b:

[0189] Compound 14a (5 g), pinacol diboronate (5.1 g), and potassium acetate (2.631 g) were added to dioxane (50 mL), followed by the addition of Pd(dppf)Cl2 (0.490 g). The reaction was carried out overnight at 85 °C under nitrogen protection. The mixture was monitored by LCMS, filtered, and the filtrate was concentrated. The concentrate was then obtained by silica gel column chromatography (PE:EA = 1:1) to give compound 14b (5.327 g). LCMS (ESI) m / z: [M+1] = 404.31.

[0190] The synthesis and preparation method of intermediate 14, steps two to five, is the same as the synthesis and preparation method of intermediate 8, steps one to four. LCMS(ESI) m / z: [M+1] = 646.73.

[0191] Synthesis of intermediate 15: 3-(4-(6-((1-(1-((((1r,4r)-4-(4-amino-3-(difluoromethyl)-1H-pyrazol-1-yl)cyclohexyl)methyl)piperidin-4-yl)oxy)pyridin-3-yl)-3-methyl-2-oxa-2,3-dihydro-1H-benzo[d]imidazol-1-yl)piperidin-2,6-dione

[0192]

[0193] The synthesis and preparation methods for intermediate 15 are the same as those for intermediate 14, steps one to five. LCMS(ESI) m / z: [M+1] = 663.37.

[0194] Synthesis of intermediate 16: 3-(5-(6-(4-(((1r,4r)-4-(4-amino-3-(difluoromethyl)-1H-pyrazol-1-yl)cyclohexyl)methyl)piperazin-1-yl)pyridin-3-yl)-3-methyl-1H-indazol-1-yl)piperidine-2,6-dione

[0195]

[0196] The synthesis and preparation methods for intermediate 16 are the same as those for intermediate 8, steps one to four. LCMS(ESI) m / z: [M+1] = 632.37.

[0197] Synthesis of intermediates 17 to 19: The preparation methods of intermediates 17 to 19 are similar to those of intermediate 16, and their numbering and structure are shown in Table 2.

[0198] Table 2: Numbering and Structure of Intermediates 17 to 19

[0199]

[0200] Synthesis of intermediate 20: 1-(7-(6-(4-(((1r,4r)-4-(4-amino-3-(difluoromethyl)-1H-pyrazol-1-yl)cyclohexyl)methyl)piperazin-1-yl)pyridin-3-yl)-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4-(1H,3H)-dione

[0201]

[0202] The synthesis and preparation methods for intermediate 20, steps one to four, are the same as those for intermediate 8. LCMS(ESI) m / z: [M+1] = 633.3.

[0203] The following are examples of the preparation of the compounds of the present invention.

[0204] Example 1: N-(3-(difluoromethyl)-1-((1r,4r)-4-((4-(5-(1-(2,6-dioxopiridine-3-yl)-3-methyl-1H-indazol-4-yl)pyridin-2-yl)piperazin-1-yl)methyl)cyclohexyl)-1H-pyrazol-4-yl)-5-morpholinopyrazolo[1,5-a]pyrimidine-3-carboxamide (TM-1)

[0205]

[0206] Compound 2 (92 mg) was added to acetonitrile (2 mL), followed by NMI (94 mg) and TCFH (121 mg). The mixture was stirred at room temperature for 15 minutes, then compound 8 (180 mg) dissolved in DMF was added, and the mixture was stirred overnight at room temperature. The mixture was monitored by LCMS, concentrated, and water was added to precipitate the solid. The solid was filtered, and compound TM-1 (37.87 mg) was prepared by high-pressure reverse phase chromatography. LCMS (ESI) m / z: [M+1] = 862.66; 1HNMR(400MHz,DMSO-d6)δ11.08(s,1H),9.40(s,1H),8.83(d,J=7.9Hz,1H),8.39(s,1H),8.29(S,1 H),8.2(d,J=2.7Hz,1H),7.66(dd,J=8.7,2.5Hz,1H),7.54(d,J=8.5Hz,1H),7.41(dd,J=8.5,7.0H z,1H),7.35–6.67(m,4H),5.80(dd,J=11.8,5.1Hz,1H),4.21(t,J=11.8Hz,1H),3.89–3.52(m,12H ),3.05–2.67(m,4H),2.36–2.12(m,6H),2.10–1.92(m,4H),1.86–1.63(m,4H),1.31–1.01(m,4H).

[0207] Example 2: 5-((1R,4R)-2-oxa-5-azabicyclo[2.2.1]heptane-5-yl)-N-(3-(difluoromethyl)-1-((1r,4R)-4-((4-(5-(1-(2,6-dioxoperidin-3-yl)-3-methyl-1H-indazol-4-yl)pyridin-2-yl)piperazin-1-yl)methyl)cyclohexyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (TM-2)

[0208]

[0209] The preparation method of compound TM-2 is the same as that of TM1 in Example 1. LCMS (ESI) m / z: [M+1] = 874.29; 1H NMR (400MHz, DMSO-d6) δ11.08(s,1H),9.51(d,J=6.3Hz,1H),8.78(d,J=7.7Hz,1H),8.40(d,J=4.3Hz,1H),8.26(d,J= 5.6Hz,1H),8.20(d,J=2.5Hz,1H),7.66(dd,J=8.7,2.5Hz,1H),7.58–7.52(m,1H),7.41(dd,J=8.5,7.0Hz,1H),7.32– 6.77(m,3H),6.45(d,J=7.8Hz,1H),5.80(dd,J=11.8,5.1Hz,1H),5.18(d,J=84.5Hz,1H),4.77(d,J=16.5Hz,1H),4.2 0(t,J=11.7Hz,1H),3.86–3.71(m,2H),3.69–3.43(m,6H),2.97–2.66(m,4H),2.40–1.58(m,15H),1.27–0.92(m,3H).

[0210] Example 3: N-(3-(difluoromethyl)-1-((1r,4r)-4-((4-(5-(1-(2,6-dioxopiridine-3-yl)-3-methyl-1H-indazol-4-yl)pyridin-2-yl)piperazin-1-yl)methyl)cyclohexyl)-1H-pyrazol-4-yl)-5-(1,4-oxazaheptane-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (TM-3)

[0211]

[0212] The preparation method of compound TM-3 is the same as that of TM1 in Example 1. LCMS (ESI) m / z: [M+1] = 876.73; 1 H NMR (400MHz, DMSO-d6) δ11.08(s,1H),9.31(s,1H),8.78(d,J=7.9Hz,1H),8.40 (s,1H),8.27(s,1H),8.21(s,1H),7.67(d,J=8.6Hz,1H),7.55(d,J=8.5Hz,1H) ,7.41(dd,J=8.5,7.0Hz,1H),7.32–6.78(m,4H),5.80(dd,J=11.8,5.1Hz,1H), 4.26–3.51(m,13H),3.05–2.60(m,5H),2.37–1.65(m,15H),1.23-1.06(m,3H).

[0213] Example 4: N-(3-(difluoromethyl)-1-((1r,4r)-4-((4-(5-(1-(2,6-dioxadiazin-3-yl)-3-methyl-1H-indazol-4-yl)pyrimidin-2-yl)piperazin-1-yl)methyl)cyclohexyl)-1H-pyrazol-4-yl)-5-morpholinopyrazolo[1,5-a]pyrimidin-3-carboxamide (TM-4)

[0214]

[0215] The preparation method of compound TM-4 is the same as that of TM1 in Example 1. LCMS (ESI) m / z: [M+1] = 863.13; 1 H NMR (400MHz, DMSO-d6) δ11.09 (s, 1H), 9.40 (s, 1H), 8.83 (d, J = 7.9Hz, 1H), 8.50 (s, 2H), 8.39(s,1H),8.29(s,1H),7.69–7.56(m,1H),7.43(dd,J=8.5,7.0Hz,1H),7.29–6.78(m ,3H),5.81(dd,J=11.8,5.2Hz,1H),4.20(t,J=8.0Hz,1H),3.87–3.86(m,12H),2.94–2. 66(m,4H),2.47–2.17(m,7H),2.10–1.91(m,4H),1.86–1.60(m,4H),1.30–1.01(m,3H).

[0216] Example 5: 5-((1R,4R)-2-oxa-5-azabicyclo[2.2.1]heptane-5-yl)-N-(3-(difluoromethyl)-1-((1r,4R)-4-((4-(5-(1-(2,6-dioxoperidin-3-yl)-3-methyl-1H-indazol-4-yl)pyrimidin-2-yl)piperazin-1-yl)methyl)cyclohexyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidin-3-carboxamide (TM-5)

[0217]

[0218] Compound TM-5 was prepared using the same method as TM1 in Example 1. LCMS (ESI) m / z: [M+1] = 875.21; 1H NMR (400MHz, DMSO-d6) δ11.09(s,1H),9.51(d,J=6.3Hz,1H),8.78(d,J=7.7Hz,1H),8.50(s,2H),8.39(d,J=4.3Hz,1H) ,8.26(d,J=5.6Hz,1H),7.58(d,J=8.6Hz,1H),7.43(dd,J=8.5,7.0Hz,1H),7.30–6.78(m,3H),6.45(d,J=7.8Hz,1H),5 .81(dd,J=11.8,5.2Hz,1H),5.28(s,0.5H),5.07(s,0.5H),4.77(d,J=17.3Hz,1H),4.27-4.16(m,1H),3.89–3.44(m,6 H),2.92-2.66(m,4H),2.50-2.42(m,4H),2.35-2.16(m,5H),2.11–1.90(m,4H),1.85-1.61(m,4H),1.30–0.99(m,3H).

[0219] Example 6: N-(3-(difluoromethyl)-1-((1r,4r)-4-((4-(5-(1-(2,6-dioxopiridine-3-yl)-3-methyl-1H-indazol-4-yl)pyrimidin-2-yl)piperazin-1-yl)methyl)cyclohexyl)-1H-pyrazol-4-yl)-5-(1,4-oxazaheptane-4-yl)pyrazolo[1,5-a]pyrimidin-3-carboxamide (TM-6)

[0220]

[0221] Compound TM-6 was prepared using the same method as TM1 in Example 1. LCMS (ESI) m / z: [M+1] = 877.43; 1H NMR (400MHz, DMSO-d6) δ11.08(s,1H),9.31(s,1H),8.78(d,J=8.0Hz,1H),8.50(s,2H),8.39(s ,1H),8.27(s,1H),7.58(dd,J=8.6,0.9Hz,1H),7.43(dd,J=8.5,7.0Hz,1H),7.31–6.93(m,2H) ,6.83(d,J=8.0Hz,1H),5.81(dd,J=11.8,5.1Hz,1H),4.44–3.54(m,12H),2.99–2.62(m,4H),2 .47-2.42(m,4H),2.31-2.15(m,5H),2.12–1.87(m,6H),1.84–1.61(m,4H),1.30–1.01(m,3H).

[0222] Example 7: N-(3-(difluoromethyl)-1-((1r,4r)-4-((4-(5-(1-(2,6-dioxoperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)pyridin-2-yl)piperazin-1-yl)methyl)cyclohexyl)-1H-pyrazol-4-yl)-5-morpholinopyrazolo[1,5-a]pyrimidine-3-carboxamide (TM-7)

[0223]

[0224] Compound TM-7 was prepared using the same method as TM1 in Example 1. LCMS (ESI) m / z: [M+1] = 878.50; 1 H NMR (400MHz, DMSO-d6) δ11.12(s,1H),9.40(s,1H),8.83(d,J=7.9Hz,1H),8.39(s,1H),8.17(d,J=2.5Hz,1H),7.69–7.54(m,1H),7.25–6. 80(m,7H),5.43(dd,J=12.7,5.3Hz,1H),4.24–4.17(m,1H),3.85–3.69(m,8H),3.60–3.49(m,4H),3.01–2.59(m,8H),2.24–0.80(m,17H).

[0225] Example 8: 5-((1R,4R)-2-oxa-5-azabicyclo[2.2.1]heptane-5-yl)-N-(3-(difluoromethyl)-1-((1r,4R)-4-((4-(5-(1-(2,6-dioxoperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)pyridin-2-yl)piperazin-1-yl)methyl)cyclohexyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (TM-8)

[0226]

[0227] The preparation method of compound TM-8 is the same as that of TM1 in Example 1. LCMS (ESI) m / z: [M+1] = 890.40; 1 H NMR (400MHz, DMSO-d6) δ11.12(s,1H),9.50(d,J=6.2Hz,1H),8.78(d,J=7.7Hz,1H),8.39(d,J=4.2Hz,1H ),8.26(d,J=5.6Hz,1H),8.17(d,J=2.4Hz,1H),7.72–7.57(m,1H),7.44–6.69(m,5H),6.45(d,J=7.7Hz, 1H),5.43(dd,J=12.7,5.4Hz,1H),5.28(s,0.5H),5.07(s,0.5H),4.77(d,J=17.3Hz,1H),4.25-4.15(m, 1H), 4.07–3.42 (m, 9H), 3.02–2.85 (m, 5H), 2.80–2.61 (m, 3H), 2.19 (d, J = 7.1Hz, 2H), 2.10–1.00 (m, 11H).

[0228] Example 9: N-(3-(difluoromethyl)-1-((1r,4r)-4-((4-(5-(1-(2,6-dioxoperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)pyridin-2-yl)piperazin-1-yl)methyl)cyclohexyl)-1H-pyrazol-4-yl)-5-(1,4-oxazaheptane-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (TM-9)

[0229]

[0230] The preparation method of compound TM-9 is the same as that of TM1 in Example 1. LCMS (ESI) m / z: [M+1] = 892.40; 1H NMR (400MHz, DMSO-d6) δ11.12(s,1H),9.31(s,1H),8.78(d,J=7.9Hz,1H),8.39(s,1H),8.16(s,1H),7.63(d,J=9.8Hz,1H),7 .53–6.36(m,6H),5.43(dd,J=12.7,5.4Hz,1H),4.28-4.14(m,1H),3.91–3.63(m,8H),3.10–2.60(m,8H),2.24–0.84(m,20H).

[0231] Example 10: N-(3-(difluoromethyl)-1-((1r,4r)-4-((4-(4-(1-(2,6-dioxopiridine-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)-1H-pyrazol-1-yl)piperidin-1-yl)methyl)cyclohexyl)-1H-pyrazol-4-yl)-5-morpholinopyrazolo[1,5-a]pyrimidine-3-carboxamide (TM-10)

[0232]

[0233] Compound 2 (59 mg) was dispersed in 4 mL of anhydrous acetonitrile, and NMI (60 mg) and TCFH (77 mg) were added. The mixture was stirred in a water bath at room temperature for 0.5 h under nitrogen protection. One drop of the sample was quenched with one drop of tetrahydropyrrole and sonicated for 30 seconds. The solution was diluted with acetonitrile and the LCMS was measured. The LCMS showed that compound C reacted completely to form the active ester. 2 mL of DMF solution of compound 11 (123 mg) was added, and the system was reacted overnight at 25–30 °C. The target molecular weight was obtained by LCMS monitoring. The solution was concentrated under reduced pressure at 50 °C to remove acetonitrile, and 4 mL of water was added. The mixture was stirred in a water bath at room temperature for 0.5 h to crystallize. The solution was filtered, washed three times with water, and a crude solid was obtained. The solid was dissolved in 2 mL of DMSO, filtered, and sent to the preparation stage to obtain compound TM-10 (53.56 mg, Y = 34%), a white solid. LCMS (ESI) m / z: [M+1] = 866.40; 1H NMR (400MHz, DMSO-d6) δ11.11(s,1H),9.40(s,1H),8.83(d,J=8.0Hz,1H),8.38(s,1H),8.29(s,1H ),8.02(s,1H),7.60(d,J=0.8Hz,1H),7.34–7.19(m,1H),7.11(d,J=7.0Hz,1H),7.04(t,J=7.8Hz, 1H),6.90(dd,J=8.1,6.5Hz,2H),5.41(dd,J=12.7,5.4Hz,1H),4.18(d,J=11.7Hz,2H),3.91–3.59 (m,8H),3.05(s,3H),3.01–2.86(m,3H),2.83–2.59(m,2H),2.35–1.43(m,15H),1.37–1.02(m,3H).

[0234] Example 11: 5-((1R,4R)-2-oxa-5-azabicyclo[2.2.1]heptane-5-yl)-N-(3-(difluoromethyl)-1-((1r,4R)-4-((4-(4-(1-(2,6-dioxoperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)-1H-pyrazol-1-yl)piperidin-1-yl)methyl)cyclohexyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (TM-11)

[0235]

[0236] The preparation method of compound TM-11 is the same as that of TM1 in Example 1. LCMS (ESI) m / z: [M+1] = 878.39; 1H NMR (400MHz, DMSO-d6) δ11.04(s,1H),9.43(d,J=6.1Hz,1H),8.71(d,J=7.7Hz,1H),8.32(d,J=4.3Hz,1H),8. 19(d,J=5.5Hz,1H),7.95(s,1H),7.54(s,1H),7.27–6.72(m,4H),6.38(d,J=7.8Hz,1H),5.35(dd,J=12.7,5. 4Hz,1H),5.21(s,0.5H),5.00(s,0.5H),4.70(dd,J=17.1,2.4Hz,1H),4.19–3.98(m,2H),3.86–3.34(m,4H), 2.99(s,3H),2.91–2.76(m,3H),2.76–2.53(m,2H),2.24–1.78(m,15H),1.77–1.45(m,4H),1.25–0.93(m,3H).

[0237] Example 12: N-(3-(difluoromethyl)-1-((1r,4r)-4-((4-(4-(1-(2,6-dioxoperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)-1H-pyrazol-1-yl)piperidin-1-yl)methyl)cyclohexyl)-1H-pyrazol-4-yl)-5-(1,4-oxaza-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (TM-12)

[0238]

[0239] Compound 4 (62 mg), anhydrous acetonitrile (4 mL), NMI (60 mg), and TCFH (77 mg) were added sequentially. The mixture was stirred in a water bath at room temperature for 0.5 h under nitrogen protection. One drop of the sample was quenched with one drop of tetrahydropyrrole and sonicated for 30 seconds. The mixture was diluted with acetonitrile and the LCMS was measured. The LCMS showed that compound C had completely reacted to form the active ester. A DMF solution of compound 11 (123 mg) in 2 mL was added, and the mixture was reacted overnight at 25–30 °C. The target molecular weight was obtained by LCMS monitoring. The mixture was concentrated under reduced pressure at 50 °C to remove acetonitrile. Water (4 mL) was added, and the mixture was stirred in a water bath at room temperature for 0.5 h to crystallize. The crystals were filtered, washed three times with water, and a crude solid was obtained. The solid was dissolved in DMSO (2 mL), filtered, and sent to the preparation stage to obtain compound TM-12 (68.88 mg), a white solid. LCMS (ESI) m / z: [M+1] = 880.39; 1H NMR (400MHz, DMSO-d6) δ11.04(s,1H),9.24(s,1H),8.71(d,J=7.9Hz,1H),8.32(s,1H),8.2 0(s,1H),8.00–7.84(m,1H),7.53(d,J=0.7Hz,1H),7.22–7.11(m,1H),7.11–7.02(m,1H),6 .97(t,J=7.8Hz,1H),6.87–6.75(m,2H),5.34(dd,J=12.7,5.4Hz,1H),4.19–3.56(m,10H), 2.98(s,3H),2.94–2.79(m,3H),2.75–2.55(m,3H),2.21–1.38(m,16H),1.30–0.91(m,3H).

[0240] Example 13: 5-((1S,4S)-2-oxa-5-azabicyclo[2.2.1]heptane-5-yl)-N-(3-(difluoromethyl)-1-((1r,4S)-4-((4-(4-(1-(2,6-dioxoperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)-1H-pyrazol-1-yl)piperidin-1-yl)methyl)cyclohexyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (TM-13)

[0241]

[0242] The preparation method of compound TM-13 is the same as that of TM11 in Example 1. LCMS (ESI) m / z: [M+1] = 878.39.

[0243] Example 14: N-(3-(difluoromethyl)-1-((1r,4r)-4-((4-((4-(1-(2,6-dioxoperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)-1H-pyrazol-1-yl)methyl)piperidin-1-yl)methyl)cyclohexyl)-1H-pyrazol-4-yl)-5-(1,4-oxaza-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (TM-14)

[0244]

[0245] Compound TM-14 was prepared using the same method as TM1 in Example 1. LCMS (ESI) m / z: [M+1] = 894.39; 1H NMR (400MHz, DMSO-d6) δ 11.11 (s, 1H), 9.30 (s, 1H), 8.76 (d, J = 7.9Hz, 1H), 8.37 (s, 1H), 8.27 (s, 1H), 7.92 (s, 1H), 7.60 (s, 1H), 7.33–6.69 (m, 3H), 5.41 (dd, J = 12.7, 5.5Hz, 1H) ),4.26–4.09(m,1H),4.09–3.62(m,10H),3.07(s,3H),3.00–2.60(m,5H),2.22–1.98(m ,5H),1.98–1.70(m,8H),1.62–1.40(m,3H),1.32–1.15(m,2H),1.02(q,J=12.6Hz,2H).

[0246] Example 15: N-(3-(difluoromethyl)-1-((1r,4r)-4-((4-((5-(1-(2,6-dioxadiazin-3-yl)-3-methyl-1H-indazol-4-yl)pyrimidin-2-yl)oxy)piperidin-1-yl)methyl)cyclohexyl)-1H-pyrazol-4-yl)-5-morpholinopyrazolo[1,5-a]pyrimidin-3-carboxamide (TM-15)

[0247]

[0248] The preparation method of compound TM-15 is the same as that of TM1 in Example 1. LCMS (ESI) m / z: [M+1] = 877.83; 1 H NMR(400MHz,DMSO-d6)δ11.10(s,1H),9.40(s,1H),8.83(d,J=7.9Hz,1H),8.82(s,2H),8 .38(s,1H),8.29(s,1H),7.65(d,J=8.6Hz,1H),7.47(dd,J=8.5,7.0Hz,1H),7.28–6.96(m ,2H),6.91(d,J=8.0Hz,1H),5.83(dd,J=11.8,5.1Hz,1H),5.11–4.97(m,1H),4.26–4.08( m,1H),3.94–3.63(m,8H),3.08–2.65(m,6H),2.36–1.51(m,18H),1.06(q,J=12.6Hz,2H).

[0249] Example 16: 5-((1R,4R)-2-oxa-5-azabicyclo[2.2.1]heptane-5-yl)-N-(3-(difluoromethyl)-1-((1r,4R)-4-((4-(5-(1-(2,6-dioxoperidin-3-yl)-3-methyl-1H-indazol-4-yl)pyrimidin-2-yl)oxy)piperidin-1-yl)methyl)cyclohexyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidin-3-carboxamide (TM-16)

[0250]

[0251] Compound TM-16 was prepared using the same method as TM1 in Example 1. LCMS (ESI) m / z: [M+1] = 890.45; 1H NMR (400MHz, DMSO-d6) δ 11.09 (s, 1H), 9.50 (d, J = 6.1Hz, 1H), 8.78 (d, J = 7.7Hz, 1H), 8.74 (s, 2H), 8.39 (d, J = 4.2Hz, 1H), 8.26 (d, J = 5.6Hz, 1H), 7.65 (d, J = 8.5Hz, 1H), 7.47 (dd, J = 8.5, 7.0Hz, 1H), 7.30–6.97 (m, 2H), 6.86 (d, J = 7 .8Hz,0.5H),6.45(d,J=7.8Hz,0.5H),5.83(dd,J=11.9,5.1Hz,1H),5.28(s,0.5H),5.13–4.99(m,1.5H),4.77(d ,J=16.5Hz,1H),4.25–4.13(m,1H),3.94–3.43(m,4H),3.02–2.61(m,6H),2.37–1.53(m,20H),1.17–1.00(m,2H).

[0252] Example 17: N-(3-(difluoromethyl)-1-((1r,4r)-4-((4-((5-(1-(2,6-dioxoperidin-3-yl)-3-methyl-1H-indazol-4-yl)pyrimidin-2-yl)oxy)piperidin-1-yl)methyl)cyclohexyl)-1H-pyrazol-4-yl)-5-(1,4-oxazaheptane-4-yl)pyrazolo[1,5-a]pyrimidin-3-carboxamide (TM-17)

[0253]

[0254] The preparation method of compound TM-17 is the same as that of TM1 in Example 1. LCMS (ESI) m / z: [M+1] = 890.45; 1H NMR (400MHz, DMSO-d6) δ11.09(s,1H),9.31(s,1H),8.78(d,J=8.0Hz,1H),8.74(s,2H),8. 39(s,1H),8.27(s,1H),7.65(d,J=8.5Hz,1H),7.47(dd,J=8.5,7.0Hz,1H),7.25–6.95(m,2 H),6.83(d,J=8.0Hz,1H),5.83(dd,J=11.9,5.1Hz,1H),5.14–4.94(m,1H),4.19(t,J=11.8 Hz,1H),4.13–3.59(m,8H),3.05–2.64(m,6H),2.41–1.55(m,20H),1.06(q,J=12.6Hz,2H).

[0255] Example 18: 5-((1R,4R)-2-oxa-5-azabicyclo[2.2.1]heptane-5-yl)-N-(3-(difluoromethyl)-1-((1r,4R)-4-((4-(5-(1-(2,6-dioxoperidin-3-yl)-3-methyl-1H-indazol-4-yl)pyridin-2-yl)oxy)piperidin-1-yl)methyl)cyclohexyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (TM-18)

[0256]

[0257] The preparation method of compound TM-18 is the same as that of TM1 in Example 1. LCMS (ESI) m / z: [M+1] = 889.89; 1H NMR (400MHz, DMSO-d6) δ11.09(s,1H),9.50(d,J=6.2Hz,1H),8.78(d,J=7.7Hz,1H),8.39(d,J=4.3Hz,1H),8.31–8.17 (m,2H),7.80(dd,J=8.5,2.5Hz,1H),7.59(dd,J=8.7,0.8Hz,1H),7.43(dd,J=8.5,7.0Hz,1H),7.32–6.96(m,2H),6.9 1–6.83(m,1.5H),6.45(d,J=7.8Hz,0.5H),5.81(dd,J=11.8,5.2Hz,1H),5.28(s,0.5H),5.12–5.03(m,1.5H),4.77(d ,J=16.6Hz,1H),4.19(t,J=11.8Hz,1H),4.00–3.43(m,4H),3.11–2.65(m,5H),2.37–1.57(m,21H),1.11–0.99(m,2H).

[0258] Example 19: N-(3-(difluoromethyl)-1-((1r,4r)-4-((4-((5-(1-(2,6-dioxadiazin-3-yl)-3-methyl-1H-indazol-4-yl)pyridin-2-yl)oxy)piperidin-1-yl)methyl)cyclohexyl)-1H-pyrazol-4-yl)-5-morpholinopyrazolo[1,5-a]pyrimidine-3-carboxamide (TM-19)

[0259]

[0260] The preparation method of compound TM-19 is the same as that of TM1 in Example 1. LCMS (ESI) m / z: [M+1] = 876.93; 1H NMR (400MHz, DMSO-d6) δ11.09(s,1H),9.40(s,1H),8.82(d,J=7.9Hz,1H),8.38(s,1H),8.29(s,1H),8 .23(dd,J=2.5,0.8Hz,1H),7.80(dd,J=8.5,2.5Hz,1H),7.59(dd,J=8.7,0.8Hz,1H),7.43(dd,J=8.5,7 .0Hz,1H),7.25–6.94(m,2H),6.93–6.86(m,2H),5.81(dd,J=11.8,5.2Hz,1H),5.07(dt,J=9.0,4.8Hz ,1H),4.25–4.10(m,1H),3.85–3.66(m,8H),3.13–2.66(m,5H),2.34–1.54(m,19H),1.17–0.97(m,2H).

[0261] Example 20: N-(3-(difluoromethyl)-1-((1r,4r)-4-((4-((5-(1-(2,6-dioxoperidin-3-yl)-3-methyl-1H-indazol-4-yl)pyridin-2-yl)oxy)piperidin-1-yl)methyl)cyclohexyl)-1H-pyrazol-4-yl)-5-(1,4-oxazaheptane-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (TM-20)

[0262]

[0263] Compound 4 (52 mg) was added to acetonitrile (2 mL), followed by NMI (50 mg) and TCFH (66 mg). The mixture was stirred at room temperature for 15 minutes. A sample was then taken, and a drop of tetrahydropyrrole was added to determine the activation value. Compound 14 (100 mg) dissolved in DMF was then added, and the mixture was stirred overnight at room temperature. The mixture was monitored by LCMS. The acetonitrile was concentrated, water was added to precipitate the solid, and the solid was filtered. Compound TM19 (18.8 mg) was prepared by high-pressure reverse phase chromatography. LCMS (ESI) m / z: [M+1] = 891.56; 1H NMR (400MHz, DMSO-d6) δ11.09(s,1H),9.31(s,1H),8.78(d,J=8.0Hz,1H),8.39(s,1H),8.27(s,1H),8.24(dd, J=2.5,0.8Hz,1H),7.80(dd,J=8.5,2.5Hz,1H),7.59(dd,J=8.6,0.9Hz,1H),7.43(dd,J=8.5,7.0Hz,1H),7.28 –6.96(m,2H),6.91–6.86(m,1H),6.83(d,J=7.9Hz,1H),5.81(dd,J=11.8,5.1Hz,1H),5.13–5.02(m,1H),4.24 –4.12(m,1H),4.10–3.53(m,8H),3.09–2.61(m,5H),2.38–1.51(m,20H),1.32–1.25(m,1H),1.12–0.99(m,2H).

[0264] Example 21: N-(3-(difluoromethyl)-1-((1r,4r)-4-((4-((5-(1-(2,6-dioxoperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)pyridin-2-yl)oxy)piperidin-1-yl)methyl)cyclohexyl)-1H-pyrazol-4-yl)5-(1,4-oxazaheptan-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (TM-21)

[0265]

[0266] The preparation method of compound TM-21 is the same as that of TM1 in Example 1. LCMS (ESI) m / z: [M+1] = 907.40; 1 H NMR (400MHz, DMSO-d6) δ11.13(s,1H),9.31(s,1H),8.78(d,J=7.9Hz,1H),8.38(s,1 H),8.27(s,1H),8.22(d,J=2.5Hz,1H),7.79(dd,J=8.5,2.5Hz,1H),7.44–6.95(m,3 H),6.96–6.79(m,3H),5.44(dd,J=12.7,5.5Hz,1H),5.12–5.01(m,1H),4.25–4.14( m,1H),4.12–3.59(m,8H),2.98–2.57(m,8H),2.31–1.54(m,18H),1.14–0.99(m,2H).

[0267] Example 22: N-(3-(difluoromethyl)-1-((1r,4r)-4-((4-((5-(1-(2,6-dioxoperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)pyridin-2-yl)oxy)piperidin-1-yl)methyl)cyclohexyl)-1H-pyrazol-4-yl)5-morpholinopyrazolo[1,5-a]pyrimidine-3-carboxamide (TM-22)

[0268]

[0269] Compound TM-22 was prepared using the same method as TM1 in Example 1. LCMS (ESI) m / z: [M+1] = 893.39; 1H NMR (400MHz, DMSO-d6) δ 11.13 (s, 1H), 9.31 (s, 1H), 8.78 (d, J = 7.9Hz, 1H), 8.38 (s, 1H), 8.27 (s, 1H), 8.22 (d, J = 2.5Hz, 1H), 7.79 (dd, J = 8.5, 2.5Hz, 1H), 7.44–6.95 (m, 3 H),6.96–6.79(m,3H),5.44(dd,J=12.7,5.5Hz,1H),5.12–5.01(m,1H),4.25–4.14( m,1H),4.12–3.59(m,8H),2.98–2.57(m,8H),2.31–1.54(m,18H),1.14–0.99(m,2H).

[0270] Example 23: 5-((1R,4R)-2-oxa-5-azabicyclo[2.2.1]heptane-5-yl)-N-(3-(difluoromethyl)-1-((1r,4R)-4-((4-(5-(1-(2,6-dioxoperidin-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-benzo[d]imidazol-4-yl)pyridin-2-yl)oxy)piperidin-1-yl)methyl)cyclohexyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-3-carboxamide (TM-23)

[0271]

[0272] Compound TM-23 was prepared using the same method as TM1 in Example 1. LCMS (ESI) m / z: [M+1] = 905.39; 1H NMR (400MHz, DMSO-d6) δ 11.08 (br, 1H), 9.50 (d, J = 6.3Hz, 1H), 8.78 (d, J = 7.7Hz, 1H), 8.39 (d, J = 4.3Hz, 1H), 8.30–8.12 (m, 2H), 7.79 (dd, J = 8.5, 2.5Hz, 1H), 7.43–6.96 (m, 3H), 6.88 (dd, J = 15.8, 8.0Hz, 2.5H), 6. 45(d,J=7.8Hz,0.5H),5.44(dd,J=12.7,5.5Hz,1H),5.28(s,0.5H),5.11–5.01(m,1.5H),4.77(d,J=16.9 Hz,1H),4.25–4.13(m,1H),3.87–3.57(m,4H),3.97–2.89(m,9H),2.30–1.27(m,17H),1.15–0.98(m,2H).

[0273] Examples 24 to 45: The preparation methods of compounds TM24 to TM45 are the same as those of TM1 in Example 1. The compound numbers and structures are shown in Table 3.

[0274] Table 3: Numbering and Structure of Compounds TM-24 to TM-45 of the Present Invention

[0275]

[0276]

[0277]

[0278] The following test examples illustrate the beneficial effects of the compounds of the present invention.

[0279] The positive compound 1 used in this invention is derived from a patent of Kymera Therapeutics, Inc., patent number WO2020113233. Its structure is as follows:

[0280]

[0281] Test Example 1: The inhibitory effect of the compound of this invention on the TNFα secretion level of human THP-1 cells

[0282] This experiment was conducted in RPMI 1640 medium containing 10% FBS / 1% penicillin / streptomycin, with an initial compound concentration of 1 μM.

[0283] Dissolve the compound DMSO to prepare a stock solution, dilute it with culture medium to a 3X working concentration, and add 100 μL to each well of a 96-well plate; count THP1 cells in the logarithmic growth phase and dilute to a 2x 10⁻⁶ concentration. 6 Add 100 μL of the above compound to each well of a 96-well plate containing 1 ng / mL of the compound, mix well, and incubate at 37°C in a 5% CO2 incubator for 1 hour. Then, continue adding LPS to the final concentration of 1 ng / mL and continue incubating at 37°C in a 5% CO2 incubator for 5 hours. Centrifuge at 1000 rpm for 1 min, and take 16 μL of the supernatant for each sample. Detect the TNFα using a TNFα ELISA kit, read the OD450 value, convert it to TNFα concentration according to the standard curve, and calculate the IC50 using GraphPad 5.0 to fit the dose-effect curve. 50 value.

[0284] Table 4. Inhibitory effects of compounds on TNFα secretion levels in human THP-1 cells.

[0285]

[0286]

[0287] The results showed that the compounds of the present invention had a significant inhibitory effect on LPS-stimulated TNFα at 100 nM-1 μM.

[0288] Test Example 2: The inhibitory effect of the compound of the present invention on IL-6 secretion levels in human peripheral blood mononuclear cells (PBMCs)

[0289] (1) Experimental method:

[0290] This experiment was conducted in RPMI 1640 medium containing 10% FBS / 1% penicillin-streptomycin, with an initial compound concentration of 1 μM. The compound was dissolved in DMSO to prepare a stock solution, which was then serially diluted 4-fold with medium to a working solution concentration of 4×. 50 μL was added to each well of a 96-well plate. PBMCs (batch number: HPP20062307, Sichuan Houpu Biotechnology Co., Ltd.) were counted and diluted to 2×10⁶ cells. 6 Add 150 μL of the above compound to each well of a 96-well plate containing 2.5 μg / mL, mix well, and incubate at 37°C in a 5% CO2 incubator for 20 h. Then, continue adding R848 to the final concentration of 2.5 μg / mL and continue incubating at 37°C in a 5% CO2 incubator for 24 h. Centrifuge at 2000 rpm for 4 min, take the supernatant, dilute 110×, and perform the assay according to the IL-6 ELISA kit. Read the OD450 value, convert it to IL-6 concentration according to the standard curve, and calculate the IC50 using GraphPad 5.0 to fit the dose-effect curve. 50 value.

[0291] (2) Experimental Results

[0292] The experimental results are shown in Table 1 below, where the EC values ​​of each compound are... 50 Values ​​are categorized according to the following descriptions:

[0293] "+" indicates IC 50 Value greater than 1 μM;

[0294] "++" indicates IC 50 Values ​​less than 1 μM and greater than 300 nM;

[0295] "++" indicates IC 50 Values ​​less than 300 nM and greater than 100 nM;

[0296] "++++" indicates IC 50 The value is less than 100 nM.

[0297] Table 5. Inhibitory effects of compounds on IL-6 secretion levels in PBMC cells.

[0298] compound <![CDATA[IC 50 (Three-way average, nM) Positive compound 1 9.58 TM-9 3.9 TM-10 1.43 TM-12 1.37 TM-21 0.34

[0299] The results showed that the compound of the present invention had a significant inhibitory effect on IL-6 secretion in PBMC cells.

[0300] Test Example 3: Study on the degradation level of IRAK4 in human peripheral blood mononuclear cells (PBMCs) by the compounds of this invention

[0301] This experiment was conducted in RPMI 1640 medium containing 10% FBS / 1% penicillin / streptomycin, with an initial compound concentration of 1 μM.

[0302] Thaw frozen human PBMCs into the culture medium. Use at least 2.5 × 10⁻⁶. 6 Cells were cultured at c / mL. PBMCs were cultured at 37°C / 5% CO2 and allowed to stand overnight. After overnight recovery, cell counting / viability was assessed using the trypan blue exclusion method. The cell density was adjusted to 2.5 × 10⁻⁶. 6 / mL. Add 90 μL to each well of a 96-well plate. Dissolve the compound in DMSO to prepare a stock solution, dilute with culture medium to a 3X working concentration, and add 10 μL to each well of the above 96-well plate. Incubate at 37°C, 5% CO2 for 20 hours. At the end of the treatment, collect the cells and centrifuge at 1800 rpm for 5 minutes. Wash with 1×PBS and centrifuge at 1800 rpm for 5 minutes. Freeze the cell aggregates and store at -80°C until further processing. Prepare the lysate by resuspending in dissolution buffer. Perform protein quantification using a BCA kit. Load 20 μg of protein per well and operate on a 26-well 4-12% Bis-Tris SDS-page gel. Transfer the PVDF membrane using the BioRad Mixed MWturbo program for 7 minutes. Block the membrane on a shaker for one hour at room temperature. Incubate the primary antibody overnight on a shaker at 4°C. Wash the membrane with 3×TBST for 5 minutes each. Secondary antibodies were added and the membrane was incubated on a shaker at room temperature for one hour. The membrane was washed with 3×TBST for 5 minutes each time and thoroughly rinsed with deionized H2O. The membrane was scanned using a Lyco Odyssey CLx scanner, and the bands were quantified using Image Studio Simplified Version 5.2 software.

[0303] Table 6 shows the degradation effects of the compounds on IRAK4 in PBMCs.

[0304] compound <![CDATA[DC 50 (nM)]]> Positive compound 1 3.19 TM-9 2.34 TM-10 1.05 TM-12 0.69 TM-21 1.58

[0305] The results show that the compound of the present invention has a significant degradation effect on IRAK4 in PBMC, and can achieve a degradation rate of more than 90%.

[0306] Test Example 4: Effect of the compound on the degradation of IRAK4 protein in THP1 cells

[0307] THP-1 cells were seeded into 96-well cell culture plates, 5 × 10⁶ cells per well. 4100 μL of culture medium per cell. Cell culture plates were incubated overnight at 37°C in a 5% CO2 incubator. 100 nmol of the prepared compound stock solution and compound-free DMSO stock solution were added to each well of cells. Cell culture plates were incubated at 37°C in a 5% CO2 incubator for 24 hours. One tablet of protease inhibitor and one tablet of phosphatase inhibitor were added to 20 mL of cell lysis buffer and gently mixed until completely dissolved. 200 mM DTT was added to 4x sample preparation buffer to prepare 4x working solution. 20x electrophoresis buffer was diluted to 1x with ultrapure water. 20x transfer buffer was diluted to 1x with ultrapure water and 20% methanol was added. 10x electrophoresis buffer was diluted to 1x with ultrapure water. IRAK4 protein primary antibody working solution: Add 20 μl of IRAK4 antibody to 20 mL of blocking buffer; β-Actin primary antibody working solution: Add 2 μl of beta-actin (13E5) Rabbit mAb antibody to 20 mL of blocking buffer. IRAK4 protein secondary antibody working solution: Donkey Anti-Goat IgG H&L (HRP) diluted 1 / 5000 in blocking buffer; β-Actin secondary antibody working solution: Anti-rabbit IgG, HRP-linked antibody diluted 1 / 10000 in blocking buffer. Centrifuge cell culture plates at 3000 rpm for 3 min, carefully aspirate most of the culture medium, invert the cell plate, centrifuge at 300 rpm for 30 s; add 45 μL of lysis buffer to each well, vortex at 300 rpm for 30 s, incubate on ice for 30 min, and pipette 30 times after 20 min; add 6 μL of 4x sample preparation solution to 18 μL of protein supernatant for sample preparation, and heat at 70℃ for 10 min. Remaining protein samples were stored at -80℃. 8 μL of sample was loaded into each well of the precast gel and electrophoresed at a constant voltage of 120V for approximately 60 min. Transfer was performed using a PVDF membrane at a constant current of 300 mA for 1 hour. After transfer, the membrane was blocked with blocking buffer at room temperature for 1 hour. The membrane was incubated overnight at 4℃ with primary antibody working solution. The membrane was washed with 1×TBST buffer for 3×10 min. The membrane was incubated with secondary antibody working solution at room temperature for 1 hour. The membrane was washed with 1×TBST buffer for 3×10 min, and then exposed to develop color. The gray values ​​of each band were calculated using ImagJ software for semi-quantitative analysis, and DC was calculated using GraphPad Prism 8.0. 50 .

[0308] "+" indicates DC 50 Value greater than or equal to 1 μM;

[0309] "++" indicates DC 50 Values ​​less than 1 μM and greater than or equal to 100 nM;

[0310] "++" indicates DC 50 Value less than 100 nM;

[0311] Table 7. Effects of compounds on IRAK4 protein degradation in THP1 cells.

[0312] compound <![CDATA[DC 50 (nM)]]> Positive compound 1 5.78 TM-2 24.28 TM-3 22.43 TM-6 17.43 TM-9 2.10 TM-12 1.27 TM-14 2.90 TM-17 16.88 TM-18 34.24 TM-20 27.58 TM-21 6.21

[0313] The results showed that the compound of the present invention had a significant degradation effect on IRAK4 in THP1 cells, and could achieve a degradation rate of more than 90%.

[0314] Test Example 5: Metabolic Stability Test

[0315] Preheat empty incubation plates T60 and NCF60 for 10 minutes; dilute human liver microsomes to 0.56 mg / mL in 100 mM phosphate buffer, transfer 445 μL of microsome working solution (0.56 mg / mL) to the preheated incubation plates T60 and NCF60, and then pre-incubate the incubation plates T60 and NCF60 with continuous shaking at 37°C for 10 minutes. Transfer 54 μL of liver microsomes to a blank plate, then add 6 μL of NAPDH cofactor to the blank plate, and then add 180 μL of quenching solution to the blank plate; add 5 μL of composite working solution (100 μM) to the incubation plates (T60 and NCF60) containing microsomes, and mix thoroughly 3 times; for the NCF60 plate, add 50 μL of buffer and mix thoroughly 3 times. Start timing; the plates will be incubated at 37°C with shaking for 60 minutes; in the quenching plate T0, add 180 μL of quenching solution and 6 μL of NAPDH cofactor. Ensure the plates are cooled to prevent evaporation; for the T60 plate, mix thoroughly three times, and immediately transfer 54 μL of the mixture to the quenching plate at time 0. Then add 44 μL of NAPDH cofactor to the incubation plate (T60). Start timing; the plates will be incubated at 37°C with shaking for 60 minutes. At 5, 15, 30, 45, and 60 minutes, add 180 μL of quenching solution to the quenching plate, mix once, and then continuously transfer 60 μL of sample from the T60 plate to the quenching plate at each time point.

[0316] For NCF60, mix once and at the 60-minute time point, transfer 60 μL of sample from the NCF60 incubator to a quenching plate containing the quenching solution. Shake all sampling plates for 10 minutes, then centrifuge at 4000 rpm for 20 minutes at 4°C. Transfer 80 μL of the supernatant to 240 μL of HPLC water and mix with a plate shaker for 10 minutes. Seal each bioanalytical plate and shake for 10 minutes before LC-MS / MS analysis.

[0317] Table 8. Metabolic stability of compounds in human liver microsomes.

[0318] compound <![CDATA[Half-life (t 1 / 2 , min)]]> Positive compound 1 39 TM-10 54 TM-12 65

[0319] The results show that the compound of this invention has considerable stability in liver microsomes, with a half-life of t. 1 / 2 It can last for more than 60 minutes.

[0320] Test Example 6: Therapeutic effect of the compound of the present invention on a psoriasis model

[0321] Male BALB / c mice, 6-8 weeks old, were randomly divided into 9 groups after one week of acclimatization: a control group, an imiquimod model group (IMQ group), an IRAK4 small molecule inhibitor PF-06650833 group (30 mg / kg and 100 mg / kg), an IRAK4 protein degradation agent positive compound group (30 mg / kg and 100 mg / kg), and the compound of this invention (10 mg / kg, 30 mg / kg, and 100 mg / kg), with 6 to 7 mice in each group. On day 0, hair was removed from the back of the mice using depilatory cream, covering an area of ​​2 × 2 cm. 2 On day one, mice were modeled using IMQ cream and treated with the corresponding drugs via gavage. The IRAK4 small molecule inhibitor PF-06650833 group (30 mg / kg and 100 mg / kg) and the compounds of this invention (15 mg / kg, 30 mg / kg, and 100 mg / kg) were administered the corresponding drugs twice daily, morning and evening, with an 8-hour interval between the two administrations. Two hours after the first gavage, IMQ cream was applied to the shaved areas on the back of the mice and the left ear, with dosages of 80 mg and 10 mg, respectively. In the positive compound 1 group, IMQ cream was applied to the shaved areas on the back of the mice and the left ear four hours after the first gavage. The blank control group and the IMQ group were administered the solvent twice daily, morning and evening, with an 8-hour interval between the two administrations. Two hours after the first gavage, petrolatum and IMQ cream were applied to the shaved areas on the back of the mice and the ears, respectively, for 7 consecutive days.

[0322] The Psoriasis Area and Severity Index (PASI) scoring criteria were used. Starting from day 1 of modeling, the psoriasis, erythema, and thickness of the skin lesions on the back were scored daily from 0 to 4, as shown in Table 1. The scores of the three items were added together to obtain the total score, which was the PASI total score. On day 7 of the experiment, typical skin of mice in each group is shown in the image below on day 7 of psoriasis modeling. Starting from day 0, the thickness of the left ear of the mice was measured daily in the morning using digital calipers, and the average value was taken.

[0323] Table 9. Psoriasis Area and Severity Index (PASI) Scoring Criteria

[0324]

[0325] The results showed that the compounds of this invention had a significant inhibitory effect on IMQ-induced psoriasis, including psoriasis, postauricular psoriasis, skin thickness, and erythema. Furthermore, they could clear more than 75% of protein degradation in the spleen and skin.

[0326] In summary, this invention discloses compounds of Formula I, which can effectively degrade IRAK4 or otherwise inhibit its activity. These compounds show great promise for use in IRAK4-mediated diseases, including immune diseases (such as psoriasis, hidradenitis suppurativa, atopic dermatitis, rheumatoid arthritis, and systemic lupus erythematosus), tumors (such as multiple myeloma, lymphocytic leukemia, and lymphoma), Alzheimer's disease, and fibrotic diseases. This provides a new option for clinical screening and / or preparation of drugs for diseases related to IRAK4 activity.

Claims

1. A compound represented by Formula III, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof: Formula III wherein, n1, n2 are independently selected from 0, 1 or 2; 2. The compound of claim 1, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: R 1 -NR 1a R 1b , 4-10 membered heterocyclyl, 5-12 membered bridged heterocyclyl, or 5-12 membered spiroheterocyclyl, wherein the heterocyclyl, bridged heterocyclyl, spiroheterocyclyl can be optionally substituted with one, two, or three R 11 substituents; R 1a , R 1b are each independently selected from hydrogen, C 1~6 1-10 alkyl, C 2~6 2-10 alkenyl, C 2~6 2-10 alkynyl, halogen-substituted C 1~6 1-10 alkyl, halogen-substituted C 2~6 2-10 alkenyl, halogen-substituted C 2~6 2-10 alkynyl, -C 0~2 alkylene-3-10 membered carbocyclyl, or -C 0~2 alkylene-4-10 membered heterocyclyl; Each R 11 Each element is independently selected from hydrogen, halogen, and C. 1~6 Alkyl, C 2~6 alkenyl, C 2~6 Alkyne- and halogen-substituted C 1~6 Alkyl, halogen-substituted C 2~6 Alkenyl, halogen-substituted C 2~6 alkynyl group, -C 0~2 Alkylene-OR 12 -C 0~2 Alkylene-NR 12 R 13 -C 0~2 Alkylene-C(O)R 12 -C 0~2 Alkylene-NR 12 C(O)R 13 -C 0~2 Alkylene-C(O)NR 12 R 13 -C 0~2 Alkylene group (-3 to 10-membered carbocyclic group or -C) 0~2 Alkylene-4 to 10-membered heterocyclic groups; R 12 , R 13 are each independently selected from hydrogen, C 1~6 alkyl, C 2~6 alkenyl, C 2~6 alkynyl, halo-substituted C 1~6 alkyl, halo-substituted C 2~6 alkenyl, halo-substituted C 2~6 alkynyl, -C 0~2 alkylene-3- to 10-membered carbocyclyl, or -C 0~2 alkylene-4- to 10-membered heterocyclyl; 3. The compound of claim 2, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: U is selected from N or CR U ; R U selected from hydrogen or C 1~6 alkyl; V is selected from a chemical bond, O, S, or CR V1 R V2 ; R V1 , R V2 are each independently selected from hydrogen or C 1~6 alkyl; The A ring is ; Q is selected from N or CR Q ; R X selected from hydrogen, halogen, cyano, C 1~6 alkyl, C 2~6 alkenyl, C 2~6 alkynyl, halogen-substituted C 1~6 alkyl, halogen-substituted C 2~6 alkenyl, halogen-substituted C 2~6 alkynyl, -C 0~2 alkylene-OR X1 , -C 0~2 alkylene-NR X1 R X2 , -C 0~2 alkylene-3-10 membered carbocyclyl or -C 0~2 alkylene-4-10 membered heterocyclyl; R X1 , R X2 are each independently selected from hydrogen, C 1~6 alkyl, C 2~6 alkenyl, C 2~6 alkynyl, halogen-substituted C 1~6 alkyl, halogen-substituted C 2~6 alkenyl or halogen-substituted C 2~6 alkynyl; R Q selected from hydrogen, halogen, C 1~6 alkyl, C 2~6 alkenyl, C 2~6 alkynyl, halogen-substituted C 1~6 alkyl, halogen-substituted C 2~6 alkenyl, or halogen-substituted C 2~6 alkynyl.

4. The compound of claim 1, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: R 1 -NR 1a R 1b , 5-membered nitrogen-containing heterocyclyl, 6-membered nitrogen-containing heterocyclyl, 7-membered nitrogen-containing heterocyclyl, 8-membered nitrogen-containing heterocyclyl, 6-membered nitrogen-containing bridged heterocyclyl, 7-membered nitrogen-containing bridged heterocyclyl, 8-membered nitrogen-containing bridged heterocyclyl, 9-membered nitrogen-containing bridged heterocyclyl, 10-membered nitrogen-containing bridged heterocyclyl, 6-membered nitrogen-containing spiroheterocyclyl, 7-membered nitrogen-containing spiroheterocyclyl, 8-membered nitrogen-containing spiroheterocyclyl, 9-membered nitrogen-containing spiroheterocyclyl, or 10-membered nitrogen-containing spiroheterocyclyl, wherein the heterocyclyl, bridged heterocyclyl, spiroheterocyclyl groups are optionally substituted with one, two, or three R 11 . n1 is selected from 0, n2 is selected from 1; or n1 is selected from 1, n2 is selected from 0; or n1 is selected from 1, n2 is selected from 1; or n1 is selected from 2, n2 is selected from 1; or n1 is selected from 1, n2 is selected from 2; R 1 selected from 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 or . U is selected from N or CH; V is selected from a bond, O or CH2; Q is selected from N or CH. The compound specifically includes:

6. Use of the compound of any one of claims 1-5, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, for the preparation of a medicament for the treatment and prevention of diseases related to or mediated by interleukin-1 receptor associated kinase 4 (IRAK4) signaling pathway, interleukin-6 (IL-6) receptor and / or tumor necrosis factor alpha (TNFa).

5. The compound of any one of claims 1-4, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: The diseases include cancer, neurodegenerative diseases, viral diseases, autoimmune diseases, inflammatory diseases, metabolic disorder diseases, diseases related to organ transplantation, immunodeficiency diseases, bone destructive diseases, infectious diseases, liver diseases, cardiovascular diseases. TM-10, TM-11, TM-12, TM-13, TM-14.

8. A pharmaceutical composition comprising a compound of any one of claims 1-5, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, in combination with a pharmaceutically acceptable adjuvant.

7. Use according to claim 6, characterized in that: ​ ​

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