An indazole amide compound and its uses

Abstract

This disclosure relates to an indazole amide compound and its uses. Specifically, this disclosure provides a compound as shown in Formula I or a pharmaceutically acceptable salt thereof. The compound of this disclosure exhibits good inhibitory activity against PDE4.

Description

Technical Field

[0001] This invention relates to the field of medicinal chemistry, specifically to an indazole amide compound and its uses. Background Technology

[0002] Phosphodiesterase 4 (PDE4) belongs to the phosphodiesterase family and is responsible for the degradation of cyclic adenosine monophosphate (cAMP) in vivo. It is the earliest discovered and most abundant protein in the phosphodiesterase family, working with nucleotide cyclases to regulate intracellular cAMP levels and playing a crucial role in cAMP signaling homeostasis. PDE4 contains four isoforms: A, B, C, and D. Isoform C is less commonly expressed in vivo, while isoforms A, B, and D are the most prevalent. Significant expression differences exist among isoforms in different tissues and organs, with isoforms B and D being the most studied. Different splicing variants of each isoform result in various splicing variants, which can be classified into long, short, ultra-short, and dead-short forms based on differences in upstream conserved regions (UCRs). Among them, the long PDE4 includes UCR1 and UCR2, the short type only includes UCR2, and the ultra-short type includes truncated UCR2. Each PDE4 subtype and splice variant may have its own unique function.

[0003] cAMP is a crucial signaling molecule that regulates important physiological and pathological processes such as inflammatory responses, fibrosis, bodily injury, and central nervous system function. It can influence the occurrence and development of inflammation, the formation of fibrosis, post-injury inflammation and repair processes, neurogenesis, the establishment of neuronal circuits, apoptosis, neuronal plasticity, sleep, sensorimotor gating, emotional stability, memory, and other cognitive functions. PDE4, as a key member regulating cAMP signaling, has received considerable attention in recent years for its application in the treatment of diseases related to inflammation, fibrosis, injury, and the central nervous system. Related diseases include inflammatory diseases such as chronic obstructive pulmonary disease (COPD), asthma, dermatitis, hepatitis, non-alcoholic fatty liver disease (NAFLD), psoriasis, rhinitis, Behçet's syndrome, arthritis, eczema, vitiligo, and ulcerative colitis; fibrotic diseases such as pulmonary fibrosis, liver fibrosis, and cystic fibrosis; injury-related diseases such as lung injury, liver injury, kidney injury, edema, and traumatic brain injury; and central nervous system diseases such as Alzheimer's disease, depression, anxiety, multiple sclerosis, stroke, and cognitive improvement.

[0004] PDE4 participates in various physiological functions and is closely related to a variety of diseases by regulating cAMP levels in the body. The main pathways in which PDE4 is involved include the cAMP / PKA / CREB signaling pathway and the MAPK / ERK / CREB pathway. By inhibiting PDE4 activity, cAMP hydrolysis is blocked, and the cellular level increases, which in turn activates cAMP-dependent protein kinase A (PKA), activating the cAMP / PKA / CREB and / or MAPK / ERK / CREB signaling pathways. This, in turn, regulates the expression of multiple target genes, including BDNF, Bax, PEPCK, and IL-2, thereby participating in a variety of physiological and pathological processes related to inflammation, fibrosis, injury, memory, and learning.

[0005] Numerous clinical and preclinical studies have demonstrated that regulating PDE4 activity and cAMP levels through inhibitors can treat a variety of diseases. Several PDE4 inhibitors are currently on the market, including roflumilast, apremilast, and claborone. Due to the gastrointestinal and central nervous system side effects of antiviral drugs, PDE4 inhibitors remain a hot research topic, with improving the therapeutic index being the primary research goal. Key research directions include altering the route of administration, discovering highly active subtype-selective inhibitors, dual-target inhibitors, and investigating indications.

[0006] The above studies indicate that PDE4 is a promising druggable target with great therapeutic potential in inflammatory diseases, fibrosis-related diseases, injury-related diseases, and central nervous system-related diseases. There is an urgent need to develop novel PDE4 inhibitors for the development of candidate drugs for these diseases. Summary of the Invention

[0007] This invention provides a compound as shown in Formula I or a pharmaceutically acceptable salt thereof.

[0008]

[0009] Among them, R 1 H, unsubstituted or R 1-1 Replacement C 1-6 Alkyl, unsubstituted or R 1-2 Substituted 3-6 membered cycloalkyl, unsubstituted or R 1-3 Substituted 3-6 membered heterocyclic alkyl groups;

[0010] R 2 For not replaced or by R 2-1 Substituted 3-6 membered cycloalkyl, unsubstituted or R 2-2 Substituted 3-6 membered heterocyclic alkyl groups, unsubstituted or R 2-3 Substituted 3-12 membered heterocyclic alkenyl groups, unsubstituted or R 2-4Replacement C 6-14 aryl, unsubstituted or R 2-5 Substituted 5-6 aryl groups;

[0011] R 3 and R 4 Each is independently H, unsubstituted, or R. 3-1 Replacement C 1-6 Alkyl, unsubstituted or R 3-2 Substituted 3-6 membered cycloalkyl, unsubstituted or R 3-3 Substituted 3-12 membered heterocyclic alkyl groups;

[0012] Or, R 3 and R 4 Together with the nitrogen atom attached to it, it forms an unsubstituted or R-shaped structure. 3-4 Substituted 3-12 membered heterocyclic alkyl groups;

[0013] R 1-1 and R 3-1 Each can be independently a halogen, hydroxyl, mercapto, amino, nitro, cyano, or C group. 1-4 Alkoxy, C 1-4 Halogenated alkoxy groups, C 1-4 Carboxyl group, -C(=O)-C 1-4 Alkyl group, -NHC(=O)-C 1-4 Alkyl or -C(=O)NH-C 1-4 alkyl;

[0014] R 1-2 R 1-3 R 2-1 R 2-2 R 2-3 R 2-4 R 2-5 R 3-2 R 3-3 and R 3-4 Each can be independently a halogen, hydroxyl, mercapto, amino, nitro, cyano, or C group. 1-4 Haloalkyl, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkoxy groups, C 1-4 Carboxyl group, -C(=O)-C 1-4 Alkyl group, -NHC(=O)-C 1-4 Alkyl or -C(=O)NH-C 1-4 alkyl.

[0015] In some embodiments, only the following substituents or groups are defined, and the definitions of substituents or groups not mentioned are the same as those described in any other embodiment (hereinafter referred to as "in some embodiments"). 1H, unsubstituted or R 1-1 Replacement C 1-6 alkyl.

[0016] In some implementation schemes, R 2 For not replaced or by R 2-3 Substituted 3-12 membered heterocyclic alkenyl groups, unsubstituted or R 2-4 Replacement C 6-14 aryl, unsubstituted or R 2-5 Substituted 5-6 aryl groups.

[0017] In some implementation schemes, R 2 For not replaced or by R 2-4 Replacement C 6-14 aryl, unsubstituted or R 2-5 Substituted 5-6 aryl groups.

[0018] In some implementation schemes, R 3 and R 4 Each is independently H, unsubstituted, or R. 3-2 Substituted 3-6 membered cycloalkyl groups.

[0019] In some implementation schemes, R 3 and R 4 Together with the nitrogen atom attached to it, it forms an unsubstituted or R-shaped structure. 3-4 Substituted 3-10 membered heterocyclic alkyl groups.

[0020] In some implementation schemes, R 3 and R 4 Together with the nitrogen atom attached to it, it forms an unsubstituted or R-shaped structure. 3-4 Substituted 4-6 membered heterocyclic alkyl groups.

[0021] In some implementation schemes, R 3 and R 4 Together with the nitrogen atom attached to it, it forms an unsubstituted or R-shaped structure. 3-4 The substituted heteroatom is a 4, 5, or 6-membered heterocyclic alkyl group selected from one or more of N, O, and C (=O), and the number of heteroatoms is 1, 2, or 3.

[0022] In some implementation schemes, R 3 and R 4 Together with the nitrogen atom attached to it, it forms an unsubstituted or R-shaped structure. 3-4 The substituted heteroatom is a 6-membered heterocyclic alkyl group selected from N and O, with 1, 2 or 3 heteroatoms.

[0023] In some implementation schemes, R 1-1 and R 3-1 Each is an independent halogen.

[0024] In some implementation schemes, R 1-1 It is a halogen.

[0025] In some implementation schemes, R 2-3 R 2-4 R 2-5 R 3-2 R 3-3 and R 3-4 Each is independently a halogen, hydroxyl, thiol, C 1-4 Haloalkyl, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-4 Carboxyl group, -C(=O)-C 1-4 alkyl.

[0026] In some implementation schemes, R 2-4 R 2-5 and R 3-4 Each is independently a halogen, hydroxyl, thiol, C 1-4 Haloalkyl, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-4 Carboxyl group, -C(=O)-C 1-4 alkyl.

[0027] In some implementation schemes, R 1 H, unsubstituted or R 1-1 Replacement C 1-6 alkyl;

[0028] R 2 For not replaced or by R 2-3 Substituted 3-12 membered heterocyclic alkenyl groups, unsubstituted or R 2-4 Replacement C 6-14 aryl, unsubstituted or R 2-5 Substituted 5-6 aryl groups;

[0029] R 3 and R 4 Each is independently H, unsubstituted, or R. 3-1 Replacement C 1-6 Alkyl, unsubstituted or R 3-2 Substituted 3-6 membered cycloalkyl, unsubstituted or R 3-3 Substituted 3-12 membered heterocyclic alkyl groups;

[0030] Or, R 3 and R 4 Together with the nitrogen atom attached to it, it forms an unsubstituted or R-shaped structure. 3-4 Substituted 3-12 membered heterocyclic alkyl groups;

[0031] R 1-1 and R 3-1 Each is an independent halogen;

[0032] R 2-3 R 2-4 R 2-5 R 3-2 R 3-3 and R 3-4 Each is independently a halogen, hydroxyl, thiol, C 1-4 Haloalkyl, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-4 Carboxyl group, -C(=O)-C 1-4 alkyl.

[0033] In some implementation schemes, R 1 H, unsubstituted or R 1-1 Replacement C 1-6 alkyl;

[0034] R 2 For not replaced or by R 2-4 Replacement C 6-14 aryl, unsubstituted or R 2-5 Substituted 5-6 aryl groups;

[0035] R 3 and R 4 Each is independently H, unsubstituted, or R. 3-2 Substituted 3-6 membered cycloalkyl, unsubstituted or R 3-3 Substituted 3-12 membered heterocyclic alkyl groups;

[0036] Or, R 3 and R 4 Together with the nitrogen atom attached to it, it forms an unsubstituted or R-shaped structure. 3-4 Substituted 3-12 membered heterocyclic alkyl groups;

[0037] R 1-1 It is a halogen;

[0038] R 2-4 R 2-5 R 3-2 R 3-3 and R 3-4 Each is independently a halogen, hydroxyl, thiol, C 1-4 Haloalkyl, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-4 Carboxyl group, -C(=O)-C 1-4 alkyl.

[0039] In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is the compound of formula I-1 or a pharmaceutically acceptable salt thereof.

[0040]

[0041] Wherein, ring A is unsubstituted or replaced by R. 3-4 Substituted 3-12 membered heterocyclic alkyl groups; R 1 R 2 and R 3-4 As defined in Equation I.

[0042] In some implementation schemes, R 1 H, unsubstituted or R 1-1 Replacement C 1-6 alkyl;

[0043] R 2 For not replaced or by R 2-3 Substituted 3-12 membered heterocyclic alkenyl groups, unsubstituted or R 2-4 Replacement C 6-14 aryl, unsubstituted or R 2-5 Substituted 5-6 aryl groups;

[0044] Ring A is unsubstituted or replaced by R 3-4 Substituted 3-12 membered heterocyclic alkyl groups;

[0045] R 1-1 It is a halogen;

[0046] R 2-3 R 2-4 R 2-5 and R 3-4 Each is independently a halogen, hydroxyl, thiol, C 1-4 Haloalkyl, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-4 Carboxyl group, -C(=O)-C 1-4 alkyl.

[0047] In some implementation schemes, R 1 H, unsubstituted or R 1-1 Replacement C 1-6 alkyl;

[0048] R 2 For not replaced or by R 2-4 Replacement C 6-14 aryl, unsubstituted or R 2-5 Substituted 5-6 aryl groups;

[0049] Ring A is unsubstituted or replaced by R 3-4 Substituted 3-12 membered heterocyclic alkyl groups;

[0050] R 1-1 It is a halogen;

[0051] R 2-4 R 2-5 and R 3-4 Each is independently a halogen, hydroxyl, thiol, C 1-4 Haloalkyl, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-4 Carboxyl group, -C(=O)-C 1-4 alkyl.

[0052] In some embodiments, the compound of formula I-1 or a pharmaceutically acceptable salt thereof is the compound of formula I-1A or a pharmaceutically acceptable salt thereof.

[0053]

[0054] Among them, rings A and R 2 As defined in Equation I-1.

[0055] In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is the compound of formula I-2 or a pharmaceutically acceptable salt thereof.

[0056]

[0057] Among them, R 3’ and R 4’ Each is independently H, unsubstituted, or R. 3-1 Replacement C 1-6 Alkyl, unsubstituted or R 3-2 Substituted 3-6 membered cycloalkyl, unsubstituted or R 3-3 Substituted 3-12 membered heterocyclic alkyl groups; R 1 R 2 R 3-1 R 3-2 and R 3-3 As defined in Equation I.

[0058] In some implementation schemes, R 1 H, unsubstituted or R 1-1 Replacement C 1-6 alkyl;

[0059] R 2 For not replaced or by R 2-3 Substituted 3-12 membered heterocyclic alkenyl groups, unsubstituted or R 2-4 Replacement C 6-14aryl, unsubstituted or R 2-5 Substituted 5-6 aryl groups;

[0060] R 3’ and R 4’ Each is independently H, unsubstituted, or R. 3-1 Replacement C 1-6 Alkyl, unsubstituted or R 3-2 Substituted 3-6 membered cycloalkyl, unsubstituted or R 3-3 Substituted 3-12 membered heterocyclic alkyl groups;

[0061] R 1-1 and R 3-1 Each is an independent halogen;

[0062] R 2-3 R 2-4 R 2-5 R 3-2 and R 3-3 Each is independently a halogen, hydroxyl, thiol, C 1-4 Haloalkyl, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-4 Carboxyl group, -C(=O)-C 1-4 alkyl.

[0063] In some implementation schemes, R 1 H, unsubstituted or R 1-1 Replacement C 1-6 alkyl;

[0064] R 2 For not replaced or by R 2-4 Replacement C 6-14 aryl, unsubstituted or R 2-5 Substituted 5-6 aryl groups;

[0065] R 3’ and R 4’ Each is independently H, unsubstituted, or R. 3-2 Substituted 3-6 membered cycloalkyl, unsubstituted or R 3-3 Substituted 3-12 membered heterocyclic alkyl groups;

[0066] R 1-1 It is a halogen;

[0067] R 2-4 R 2-5 R 3-2 and R 3-3 Each is independently a halogen, hydroxyl, thiol, C 1-4 Haloalkyl, C 1-4 Alkyl, C1-4 Alkoxy, C 1-4 Haloalkyl, C 1-4 Carboxyl group, -C(=O)-C 1-4 alkyl.

[0068] In some embodiments, the compound of formula I-2 or a pharmaceutically acceptable salt thereof is the compound of formula I-2A or a pharmaceutically acceptable salt thereof.

[0069]

[0070] Among them, R 2 R 3’ and R 4’ As defined in Equation I-2.

[0071] In some embodiments, the compound of formula I or a pharmaceutically acceptable salt thereof is the compound of formulas I-3 or a pharmaceutically acceptable salt thereof.

[0072]

[0073] Among them, R 2 R 3 and R 4 As defined in Equation I.

[0074] In some implementation schemes, R 2 For not replaced or by R 2-3 Substituted 3-12 membered heterocyclic alkenyl groups, unsubstituted or R 2-4 Replacement C 6-14 aryl, unsubstituted or R 2-5 Substituted 5-6 aryl groups;

[0075] R 3 and R 4 Each is independently H, unsubstituted, or R. 3-1 Replacement C 1-6 Alkyl, unsubstituted or R 3-2 Substituted 3-6 membered cycloalkyl, unsubstituted or R 3-3 Substituted 3-12 membered heterocyclic alkyl groups;

[0076] Or, R 3 and R 4 Together with the nitrogen atom attached to it, it forms an unsubstituted or R-shaped structure. 3-4 Substituted 3-12 membered heterocyclic alkyl groups;

[0077] R 3-1 It is a halogen;

[0078] R 2-3 R 2-4 R 2-5 R 3-2R 3-3 and R 3-4 Each is independently a halogen, hydroxyl, thiol, C 1-4 Haloalkyl, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-4 Carboxyl group, -C(=O)-C 1-4 alkyl.

[0079] In some implementation schemes, R 2 For not replaced or by R 2-4 Replacement C 6-14 aryl, unsubstituted or R 2-5 Substituted 5-6 aryl groups;

[0080] R 3 and R 4 Each is independently H, unsubstituted, or R. 3-2 Substituted 3-6 membered cycloalkyl, unsubstituted or R 3-3 Substituted 3-12 membered heterocyclic alkyl groups;

[0081] Or, R 3 and R 4 Together with the nitrogen atom attached to it, it forms an unsubstituted or R-shaped structure. 3-4 Substituted 3-12 membered heterocyclic alkyl groups;

[0082] R 3-1 It is a halogen;

[0083] R 2-4 R 2-5 R 3-2 R 3-3 and R 3-4 Each is independently a halogen, hydroxyl, thiol, C 1-4 Haloalkyl, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-4 Carboxyl group, -C(=O)-C 1-4 alkyl.

[0084] In some implementation schemes, R 1 R 3 R 4 R 3’ R 4’ R 1-2 R 1-3 R 2-1 R 2-2 R 2-3 R 2-4 R 2-5 R3-2 R 3-3 and R 3-4 In, the C 1-6 Each alkyl group is independently C10. 1-4 Alkyl groups, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl. In some embodiments, R 1 In, the C 1-6 The alkyl group is ethyl.

[0085] In some implementation schemes, R 1 R 2 R 3 and R 4 In this context, each of the 3-6 membered cycloalkyl groups is independently cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.

[0086] In some implementation schemes, R 1 and R 2 In this context, each of the 3-6 membered heterocyclic alkyl groups is independently a 3-6 membered heterocyclic alkyl group whose heteroatoms are selected from one or more of N, O and S, and whose number of heteroatoms is 1, 2 or 3.

[0087] In some implementation schemes, R 2 In some embodiments, the 3-12 membered heterocyclic alkenyl group is a 3-12 membered heterocyclic alkenyl group in which "the heteroatoms are selected from one or more of N, O, S, and C (=O), and the number of heteroatoms is 1, 2, or 3". In some embodiments, the heterocyclic alkenyl group is a 5-12 membered heterocyclic alkenyl group in which "the heteroatoms are selected from one or more of N, O, S, and C (=O), and the number of heteroatoms is 1, 2, or 3". In some embodiments, the 3-12 membered heterocyclic alkenyl group is a 5-12 membered heterocyclic alkenyl group in which "the heteroatoms are selected from one or more of O, S, and C (=O), and the number of heteroatoms is 1, 2, or 3", and has 1-2 carbon-carbon double bonds. In some embodiments, R 2 In this context, the 3-12 membered heterocyclic alkenyl group is...

[0088] In some implementation schemes, R 2 In, the C 6-14 The aryl group is phenyl or naphthyl. In some embodiments, the C... 6-14 The aryl group is phenyl.

[0089] In some implementation schemes, R 2In this context, the 5-6 membered heteroaryl group is a 5-6 membered heteroaryl group whose heteroatoms are selected from one or more of N, O, and S, and the number of heteroatoms is 1, 2, or 3. In some embodiments, the 5-6 membered heteroaryl group includes, but is not limited to, pyrroleyl, furanyl, thiophenyl, pyridyl, or pyranyl. In some embodiments, the 5-6 membered heteroaryl group is thiophenyl or pyridyl.

[0090] In some implementation schemes, R 3 In some embodiments, the 3-12 membered heterocyclic alkyl group is a 3-12 membered heterocyclic alkyl group in which "the heteroatoms are selected from one or more of N, O, S, C(=O) and S(=O)2, and the number of heteroatoms is 1, 2 or 3". In some embodiments, the 3-12 membered heterocyclic alkyl group is a 3-10 membered heterocyclic alkyl group in which "the heteroatoms are selected from one or more of N, O, S, C(=O) and S(=O)2, and the number of heteroatoms is 1, 2 or 3". In some embodiments, the 3-12 membered heterocyclic alkyl group is a 3-10 membered heterocyclic alkyl group in which "the heteroatoms are selected from one or more of O, C(=O) and S(=O)2, and the number of heteroatoms is 1, 2 or 3". In some embodiments, the 3-12 membered heterocyclic alkyl group is...

[0091] In some implementations, when R 3 and R 4 Together with the nitrogen atom attached to it, it forms an unsubstituted or R-shaped structure. 3-4 When a 3-12-membered heterocyclic alkyl group is substituted, the 3-12-membered heterocyclic alkyl group is a 3-12-membered heterocyclic alkyl group in which "the heteroatoms are selected from one or more of N, O, S, C(=O), and S(=O)2, and the number of heteroatoms is 1, 2, or 3". In some embodiments, the 3-12-membered heterocyclic alkyl group is a 3-10-membered heterocyclic alkyl group in which "the heteroatoms are selected from one or more of N, O, and C(=O), and the number of heteroatoms is 1, 2, or 3". In some embodiments, the 3-12-membered heterocyclic alkyl group is a 4, 5, or 6-membered heterocyclic alkyl group in which "the heteroatoms are selected from one or more of N, O, and C(=O), and the number of heteroatoms is 1, 2, or 3". In some embodiments, the 3-12-membered heterocyclic alkyl group is an azacyclic butane, morpholino, pyrrolidone, piperidinyl, piperazine, etc.

[0092]

[0093] In some embodiments, ring A is a 3-12 membered heterocyclic alkyl group, wherein the heteroatom is selected from one or more of N, O, S, C(=O), and S(=O)₂, and the number of heteroatoms is 1, 2, or 3. In some embodiments, ring A is a 3-10 membered heterocyclic alkyl group, wherein the heteroatom is selected from one or more of N, O, and C(=O), and the number of heteroatoms is 1, 2, or 3. In some embodiments, the 3-12 membered heterocyclic alkyl group is a 4, 5, or 6 membered heterocyclic alkyl group, wherein the heteroatom is selected from one or more of N, O, and C(=O), and the number of heteroatoms is 1, 2, or 3. In some embodiments, ring A is an azahexacyclic butane, morpholino, pyrrolidone, piperidinyl, piperazine, etc.

[0094] In some implementation schemes, R 1-1 R 1-2 R 1-3 R 2-1 R 2-2 R 2-3 R 2-4 R 2-5 R 3-1 R 3-2 R 3-3 and R 3-4 The number of each is independently one or more. In some implementations, R 1-1 R 1-2 R 1-3 R 2-1 R 2-2 R 2-3 R 2-4 R 2-5 R 3 -1 R 3-2 R 3-3 and R 3-4 The number of each is independently 1, 2 or 3.

[0095] In some implementation schemes, R 1-1 R 1-2 R 1-3 R 2-1 R 2-2 R 2-3 R 2-4 R 2-5 R 3-1 R 3-2 R 3-3 and R 3-4 In this embodiment, each of the halogens is independently fluorine, chlorine, or bromine. In some embodiments, the halogen is fluorine or chlorine.

[0096] In some implementation schemes, R1-1 R 1-2 R 1-3 R 2-1 R 2-2 R 2-3 R 2-4 R 2-5 R 3-1 R 3-2 R 3-3 and R 3-4 In, the C 1-4 Each alkyl group is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl.

[0097] In some implementation schemes, R 1-1 R 1-2 R 1-3 R 2-1 R 2-2 R 2-3 R 2-4 R 2-5 R 3-1 R 3-2 R 3-3 and R 3-4 In, the C 1-4 Each alkoxy group is independently methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, or tert-butoxy. In some embodiments, the C 1-4 The alkoxy group is a methoxy group.

[0098] In some implementation schemes, R 1-2 R 1-3 R 2-1 R 2-2 R 2-3 R 2-4 R 2-5 R 3-2 R 3-3 and R 3-4 In, the C 1-4 The haloalkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl, substituted with one, two, or three fluorine, chlorine, or bromine atoms. In some embodiments, the C... 1-4 The alkyl halide is -CH2F, -CHF2 or -CF3.

[0099] In some implementation schemes, R 1-1 R 1-2 R 1-3 R 2-1 R 2-2 R 2-3 R 2-4 R 2-5 R3-1 R 3-2 R 3-3 and R 3-4 In, the C 1-4 The haloalkoxy group is a methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, or tert-butoxy group substituted with one, two, or three fluorine, chlorine, or bromine atoms. In some embodiments, the C... 1-4 The haloalkoxy group is a methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy, or tert-butoxy group substituted with 1, 2, or 3 fluorine atoms.

[0100] In some embodiments, the compound represented by Formula I or a pharmaceutically acceptable salt thereof is any of the following structural compounds or pharmaceutically acceptable salts thereof.

[0101]

[0102]

[0103]

[0104] This disclosure provides a method for preparing the compound shown in Formula I, comprising: performing a condensation reaction between the compound shown in Formula II and the compound shown in Formula III to obtain the compound shown in Formula I.

[0105]

[0106] Among them, R 1 R 2 R 3 and R 4 As defined in the compound shown in Formula I above.

[0107] In some embodiments, the conditions and operations of the condensation reaction may be those conventional for such reactions in the art.

[0108] This disclosure provides a pharmaceutical composition comprising the compound shown in Formula I above or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0109] This disclosure also provides the use of the above-described compound of Formula I or a pharmaceutically acceptable salt thereof or the above-described pharmaceutical composition in the preparation of a medicament as a PDE4 inhibitor. In some embodiments, the PDE4 inhibitor is a PDE4B and / or a PDE4D inhibitor.

[0110] This disclosure also provides the use of the above-described compound of Formula I or a pharmaceutically acceptable salt thereof or the above-described pharmaceutical composition in the preparation of a medicament for the prevention and / or treatment of PDE4-related diseases. In some embodiments, the PDE4 is PDE4B and / or PDE4D. In some embodiments, the PDE4-related diseases are inflammatory diseases, fibrotic diseases, traumatic diseases, and central nervous system diseases. In some embodiments, the inflammatory diseases include chronic obstructive pulmonary disease, asthma, dermatitis, hepatitis, non-alcoholic fatty liver disease, psoriasis, rhinitis, Behçet's syndrome, arthritis, eczema, vitiligo, and ulcerative colitis. In some embodiments, the fibrotic diseases include pulmonary fibrosis, liver fibrosis, and cystic fibrosis. In some embodiments, the traumatic diseases include lung injury, liver injury, kidney injury, edema, and traumatic brain injury. In some embodiments, the central nervous system diseases include Alzheimer's disease, antidepressants, anti-anxiety medications, multiple sclerosis, stroke, and cognitive impairment-related diseases.

[0111] This disclosure also provides the use of the above-described compounds of Formula I or their pharmaceutically acceptable salts, or the above-described pharmaceutical compositions, in the preparation of medicaments for the prevention and / or treatment of inflammatory diseases, fibrotic diseases, traumatic diseases, and central nervous system diseases. In some embodiments, the inflammatory diseases include chronic obstructive pulmonary disease, asthma, dermatitis, hepatitis, non-alcoholic fatty liver disease, psoriasis, rhinitis, Behçet's syndrome, arthritis, eczema, vitiligo, and ulcerative colitis. In some embodiments, the fibrotic diseases include pulmonary fibrosis, liver fibrosis, and cystic fibrosis. In some embodiments, the traumatic diseases include lung injury, liver injury, kidney injury, edema, and traumatic brain injury. In some embodiments, the central nervous system diseases include Alzheimer's disease, antidepressants, anti-anxiety medications, multiple sclerosis, stroke, and cognitive enhancement.

[0112] The terminology is defined as follows.

[0113] On the other hand, without specifying a particular configuration in this disclosure, the compounds disclosed herein may exist in specific geometric or stereoisomeric forms. This disclosure envisions all such compounds, including cis and trans isomers, (-)- and (+)- enantiomers, (R)- and (S)- enantiomers, diastereomers, (D)- isomers, (L)- isomers, and racemic mixtures thereof, as well as other mixtures, such as mixtures enriched with enantiomers or diastereomers, all of which are within the scope of this disclosure. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are included within the scope of this disclosure.

[0114] Furthermore, the compounds and intermediates of this disclosure may also exist in different tautomer forms, and all such forms are included within the scope of this disclosure. The terms "tautomer" or "tautomer form" refer to structural isomers with different energies that can interconvert via low energy barriers. For example, proton tautomers (also known as proton transfer tautomers) include interconversions via proton transfer, such as keto-enol and imine-enamine, lactam-lactamimide isomerization. An example of a lactam-lactamimide equilibrium is between A and B as shown below.

[0115]

[0116] All compounds in this disclosure can be classified as type A or type B. All tautomers are within the scope of this disclosure. The nomenclature of compounds does not exclude any tautomers.

[0117] The compounds disclosed herein may be asymmetric, for example, having one or more stereoisomers. Unless otherwise stated, all stereoisomers include, for example, enantiomers and diastereomers. The compounds containing asymmetric carbon atoms of this disclosure can be isolated in optically active pure form or in racemic form. The optically active pure form can be resolved from racemic mixtures or synthesized using chiral starting materials or chiral reagents.

[0118] Optically active (R)- and (S)- isomers, as well as D- and L- isomers, can be prepared by chiral synthesis, chiral reagents, or other conventional techniques. To obtain an enantiomer of a compound disclosed herein, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide a pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a salt of the diastereomeric isomer is formed with a suitable optically active acid or base, followed by diastereomeric resolution using conventional methods known in the art, and then the pure enantiomer is recovered. Furthermore, the separation of enantiomers and diastereomeric isomers is typically accomplished by using chromatography employing a chiral stationary phase and optionally combined with chemical derivatization (e.g., from amines to carbamates).

[0119] The term "alkyl" refers to a saturated aliphatic hydrocarbon group, which is a straight-chain or branched group containing 1 to 30 carbon atoms, preferably an alkyl group containing 1 to 12 carbon atoms, more preferably an alkyl group containing 1 to 10 carbon atoms, more preferably an alkyl group containing 1 to 6 carbon atoms, and even more preferably an alkyl group containing 1 to 4 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2, 3-Dimethylpentyl, 2,4-Dimethylpentyl, 2,2-Dimethylpentyl, 3,3-Dimethylpentyl, 2-Ethylpentyl, 3-Ethylpentyl, n-Octyl, 2,3-Dimethylhexyl, 2,4-Dimethylhexyl, 2,5-Dimethylhexyl, 2,2-Dimethylhexyl, 3,3-Dimethylhexyl, 4,4-Dimethylhexyl, 2-Ethylhexyl, 3-Ethylhexyl, 4-Ethylhexyl, 2-Methyl-2-Ethylpentyl, 2-Methyl-3-Ethylpentyl, n-Nonyl, 2-Methyl-2-Ethylhexyl, 2-Methyl-3-Ethylhexyl, 2,2-Diethylpentyl, n-Decyl, 3,3-Diethylhexyl, 2,2-Diethylhexyl, and their various branched isomers, etc. More preferably, alkyl groups containing 1 to 6 carbon atoms are used. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, etc.

[0120] The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, wherein the cycloalkyl ring contains 3 to 20 carbon atoms, preferably 3 to 7 carbon atoms. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, etc.; polycyclic cycloalkyls include spirocyclic, fused-ring, and bridged-ring cycloalkyls.

[0121] The term "heterocyclic alkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent containing 3 to 20 ring atoms, one or more of which are selected from nitrogen, oxygen, C (=O), or S (=O). m (where m is an integer from 0 to 2) heteroatoms, but excluding the ring portion of -OO-, -OS-, or -SS-, with the remaining ring atoms being carbon. Preferably, it contains 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; more preferably, it contains 3 to 7 ring atoms. Non-limiting examples of monocyclic heterocyclic alkyl groups include pyrrolidinyl, imidazolyl, tetrahydrofuranyl, tetrahydrothiophenyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, etc. Polycyclic heterocyclic alkyl groups include spirocyclic, fused-ring, and bridged-ring heterocyclic alkyl groups. Non-limiting examples of "heterocyclic alkyl" include:

[0122]

[0123] etc.

[0124] The heterocyclic alkyl ring may be fused to an aryl or heteroaryl ring, wherein the ring attached to the parent structure is a heterocyclic alkyl ring, and non-limiting examples include:

[0125] wait.

[0126] The term "aryl" refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (i.e., a ring sharing adjacent carbon atom pairs) group having a conjugated π-electron system, preferably 6- to 12-membered, such as phenyl and naphthyl. The aryl ring may be fused to a heteroaryl, heterocycloalkyl, or cycloalkyl ring, wherein the ring attached to the parent structure is an aryl ring, and non-limiting examples include:

[0127]

[0128] The term "heteroaryl" refers to a heteroaryl system comprising 1 to 4 heteroatoms and 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur, and nitrogen. The heteroaryl group is preferably 5 to 12-membered, more preferably 5- or 6-membered. For example, non-limiting examples include: imidazolyl, furanyl, thiophene, thiazolyl, pyrazolyl, oxazolyl, isoxazolyl, pyrrole, tetrazolyl, pyridinyl, pyrimidinyl, thiadiazole, pyrazinyl, triazolyl, indazole, benzimidazolyl, etc. wait.

[0129] The heteroaryl ring may be fused to an aryl, heterocyclic alkyl, or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring, and non-limiting examples include:

[0130]

[0131] The term "spiroring" refers to a compound in which two rings share a single atom.

[0132] The term "spirocycloalkyl" refers to a polycyclic group consisting of 5 to 20 quintile rings sharing a single carbon atom (called a spiro atom), which may contain one or more double bonds, but none of the rings has a fully conjugated π-electron system. Preferably, it is 6 to 14 quintiles, more preferably 7 to 10 quintiles. Spirocycloalkyl groups are classified into monospirocycloalkyl, bispirocycloalkyl, or polyspirocycloalkyl groups based on the number of shared spiro atoms between the rings, with monospirocycloalkyl and bispirocycloalkyl groups being preferred. More preferably, it is a 4-quintile, 4-quintile, 4-quintile, 5-quintile, or 5-quintile / 6-quintile monospirocycloalkyl group. "Spirocarbon ring" refers to the ring system within the spirocycloalkyl group. Non-limiting examples of spirocycloalkyl groups include:

[0133]

[0134] The term "spiroheterocyclic group" refers to a polycyclic heterocyclic group consisting of 5 to 20 member monocyclic rings sharing a single atom (called a spiro atom), wherein one or more ring atoms are selected from nitrogen, oxygen, or S(O). m The rings consist of heteroatoms (where m is an integer from 0 to 2), with the remaining ring atoms being carbon. They may contain one or more double bonds, but none of the rings has a fully conjugated π-electron system. Preferably, they are 6 to 14 fused, more preferably 7 to 10 fused. Spiroheterocyclic groups are classified into monospirocyclic, bispirocyclic, or multispirocyclic groups based on the number of shared spiroatoms between rings, with monospirocyclic and bispirocyclic groups being preferred. More preferably, they are 4-membered / 4-membered, 4-membered / 5-membered, 4-membered / 6-membered, 5-membered / 5-membered, or 5-membered / 6-membered monospirocyclic groups. "Spiroheterocyclic" refers to the ring system within the spirocyclic group. Non-limiting examples of spirocyclic groups include:

[0135]

[0136] The term "fused ring" refers to a compound in which two or more rings are fused together by sharing two adjacent atoms.

[0137] The term "fused-ring alkyl" refers to a 5- to 20-membered polycyclic carbon group in which each ring in the system shares an adjacent pair of carbon atoms with other rings in the system, wherein one or more rings may contain one or more double bonds, but no ring has a fully conjugated π-electron system. Preferably, it is 6- to 14-membered, more preferably 7- to 10-membered. Depending on the number of constituent rings, it can be classified as bicyclic, tricyclic, tetracyclic, or polycyclic fused-ring alkyl, preferably bicyclic or tricyclic, more preferably 5-membered / 5-membered or 5-membered / 6-membered bicyclic alkyl. Non-limiting examples of fused-ring alkyl include:

[0138]

[0139] The term "fused heterocyclic group" refers to a 5- to 20-membered polycyclic heterocyclic group in which each ring in the system shares an adjacent pair of atoms with the other rings in the system. One or more rings may contain one or more double bonds, but none of the rings has a fully conjugated π-electron system. One or more ring atoms are selected from nitrogen, oxygen, or S(O). m (where m is an integer from 0 to 2) heteroatoms, with the remaining ring atoms being carbon. Preferably, it consists of 6 to 14 rings, more preferably 7 to 10 rings. Depending on the number of rings, it can be classified as a bicyclic, tricyclic, tetracyclic, or polycyclic fused heterocyclic group, preferably bicyclic or tricyclic, more preferably 5-membered / 5-membered or 5-membered / 6-membered bicyclic fused heterocyclic group. "Fused heterocyclic" refers to the ring system in the fused heterocyclic group. Non-limiting examples of fused heterocyclic groups include:

[0140]

[0141] The term "fused aryl" can refer to an unsaturated aromatic fused ring structure containing 5-14 ring atoms (including at least one heteroatom), formed by two or more ring structures sharing two adjacent atoms, and includes cases where carbon, nitrogen, and sulfur atoms can be substituted with oxygen. Preferred terms include "5-12 fused aryl", "7-12 fused aryl", "9-12 fused aryl", etc., such as benzofuranyl, benzoisofuranyl, benzothiopheneyl, indolyl, isoindolyl, benzoxazolyl, benzoimidazolyl, indazole, benzotriazolyl, quinolinyl, 2-quinolinone, 4-quinolinone, 1-isoquinolinone, isoquinolinyl, acridineyl, phenanthidineyl, benzopyridinyl, phthalazinyl, quinazolinyl, quinoxalazinyl, quinoxalazinyl, quinoxalazinyl, phenazinyl, pteridineyl, purineyl, naphthidineyl, phenazinyl, phenothiazinyl, etc. "Dense aromatic rings" refers to the ring system in dense aromatic groups.

[0142] The term "bridged ring" refers to a structure formed by two or more ring structures sharing two non-adjacent ring atoms.

[0143] The term "bridged cycloalkyl" refers to a 5- to 20-membered polycyclic carbon group in which any two rings share two non-directly bonded carbon atoms. It may contain one or more double bonds, but none of the rings has a fully conjugated π-electron system. Preferably, it is 6- to 14-membered, more preferably 7- to 10-membered. Depending on the number of rings, it can be classified as bicyclic, tricyclic, tetracyclic, or polycyclic bridged cycloalkyl, preferably bicyclic, tricyclic, or tetracyclic, and more preferably bicyclic or tricyclic. Non-limiting examples of bridged cycloalkyl groups include:

[0144]

[0145] The term "bridged heterocyclic group" refers to a 5- to 14-membered polycyclic heterocyclic group in which any two rings share two non-directly bonded atoms. It may contain one or more double bonds, but none of the rings has a fully conjugated π-electron system. One or more ring atoms are selected from nitrogen, oxygen, or S(O). m (where m is an integer from 0 to 2) heteroatoms, with the remaining ring atoms being carbon. Preferably, it consists of 6 to 14 rings, more preferably 7 to 10 rings. Depending on the number of rings, it can be classified as a bicyclic, tricyclic, tetracyclic, or polycyclic bridged heterocyclic group, preferably bicyclic, tricyclic, or tetracyclic, and more preferably bicyclic or tricyclic. Non-limiting examples of bridged heterocyclic groups include:

[0146]

[0147] The term "alkoxy" refers to -O- (alkyl), where alkyl is defined as described above.

[0148] The term "hydroxyl group" refers to -OH.

[0149] The term "thiol" refers to -SH.

[0150] The term "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0151] The term "halogenated alkyl" refers to an alkyl group that has been substituted with a halogen, wherein the alkyl group is as defined above.

[0152] The term "cyano" refers to -CN.

[0153] The term "nitro" refers to -NO2.

[0154] The term "amino" refers to -NH2.

[0155] The term "carboxyl group" refers to -C(O)OH.

[0156] The term "substituted" refers to one or more hydrogen atoms in a group, preferably up to five, more preferably one to three hydrogen atoms, which are independently substituted by the corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art can determine possible or impossible substitutions without much effort (through experiment or theory).

[0157] "Being replaced by one or more..." means that it can be replaced by a single or multiple substituents. When replaced by multiple substituents, it can be a plurality of identical substituents or a combination of one or a plurality of different substituents.

[0158] The term "connection," when referring to the link between two molecules, means that the two molecules are connected by a covalent bond or by a non-covalent bond (e.g., a hydrogen bond or an ionic bond), including direct and indirect connections.

[0159] The term "direct link" refers to the connection between a first compound or group and a second compound or group without any intercalating atoms or groups. The term "indirect link" refers to the connection between a first compound or group and a second compound or group via an intermediate group, compound, or molecule (e.g., a linking group).

[0160] In the chemical structure of the compounds described in this disclosure, the bonds... This indicates that the configuration is not specified; that is, if chiral isomers exist in the chemical structure, the bond... It can be or Or simultaneously include and Two configurations. Although all the above structural formulas are shown in certain isomer forms for simplicity, this disclosure can include all isomers, such as tautomers, rotatimers, geometric isomers, diastereomers, racemates, and enantiomers. In the chemical structure of the compounds described in this disclosure, the bonds... No configuration was specified, i.e., key The configuration can be E-type or Z-type, or it can contain both E-type and Z-type configurations.

[0161] Unless otherwise specified, the symbols used in this article are as follows: This indicates that it can be connected with one or more groups according to the scope of disclosure described herein.

[0162] In this disclosure, the terms “comprising” or “including” may be replaced with “consisting of”.

[0163] The term "composition" refers to a mixture of a drug containing one or more of the compounds described herein or their physiologically pharmaceutically acceptable salts or prodrugs, along with other chemical components, such as physiologically pharmaceutically acceptable carriers and excipients. The purpose of the composition is to facilitate administration to a living organism, thereby promoting the absorption of the active ingredient and its biological activity.

[0164] The terms “pharmaceutical-grade excipient” or “pharmaceutical-acceptable excipient” include, but are not limited to, any adjuvant, carrier, excipient, flow aid, sweetener, diluent, preservative, dye / coloring agent, flavoring agent, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier that has been approved by the U.S. Food and Drug Administration for use in humans or livestock.

[0165] Unless otherwise specified, the "compounds" disclosed herein may exist independently as salts, mixed salts, or non-salts (e.g., free acids or free bases). When present as salts or mixed salts, they may be pharmaceutically acceptable or medicinally usable salts.

[0166] The terms “pharmaceutically acceptable salt” and “medicinal salt” are used interchangeably to refer to both pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts.

[0167] "Pharmaceutically acceptable acid addition salts" refer to salts formed with inorganic or organic acids that retain the bioavailability of the free base without other side effects and can be prepared by methods known in the art.

[0168] "Pharmaceutically acceptable base addition salts" refer to salts formed with inorganic or organic bases that retain the bioavailability of the free acid without other side effects, and these salts can be prepared by methods known in the art.

[0169] "Effective amount," "effective dose," "effective therapeutic amount," or "therapeutic effective amount" means the amount of a drug, compound, or pharmaceutical composition necessary to achieve any one or more beneficial or desired therapeutic outcome. For preventative use, beneficial or desired outcomes include eliminating or reducing risk, mitigating severity, or delaying the onset of a condition, including the condition itself, its complications, and the biochemical, histological, and / or behavioral symptoms of intermediate pathological phenotypes that occur during the development of the condition.

[0170] As used herein, the terms “subject,” “patient,” “subject,” or “individual” are used interchangeably and include human or non-human animals, such as mammals, such as humans or monkeys. Attached Figure Description

[0171] Figure 1 The compound in this example inhibits LPS-induced expression of the inflammatory factor TNF-α in BV2 cells. Detailed Implementation Plan

[0172] The present disclosure is further described below with reference to embodiments, but these embodiments are not intended to limit the scope of the disclosure. Experimental methods in the embodiments of this disclosure that do not specify specific conditions are generally performed under conventional conditions or as recommended by the raw material or product manufacturer. Reagents whose source is not specified can be obtained from any supplier of molecular biology reagents at the quality / purity required for molecular biology applications.

[0173] Unless otherwise specified, all reagents used in the following examples are commercially available products.

[0174] Example 1

[0175] Preparation of (3-(5-chlorothiophene-2-yl)-1-(2,2-difluoroethyl)-1H-indazole-5-yl)(3,3-difluoroazacyclobutane-1-yl) methyl ketone (1)

[0176] Step 1. Synthesis of methyl 3-bromo-1H-indazole-5-carboxylic acid

[0177]

[0178] 1.46 g of methyl 1H-indazole-5-carboxylate (8.30 mmol) was dissolved in 30 mL of DMF, and 1.77 g of NBS (9.96 mmol) was added. The mixture was heated at 85 °C for 12 h. The reaction was monitored by TLC. After the reaction was completed, a large amount of water was added to quench the reaction. The mixture was extracted three times with ethyl acetate (30 mL × 3). The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate for 30 min, and purified by column chromatography (petroleum ether: ethyl acetate = 8:1) to give 1.74 g of white flaky solid, with a yield of 82%.

[0179] 1 H NMR (300MHz, DMSO-d6) δ (ppm) 13.81 (s, 1H), 8.19 (dd, J = 1.5, 0.8Hz, 1H), 8.00 (dd, J = 8.8, 1.6Hz, 1H), 7.68 (dd, J = 8.8, 0.8Hz, 1H), 3.88 (s, 3H).

[0180] Step 2. Synthesis of methyl 3-bromo-1-(2,2-difluoroethyl)-1H-indazole-5-carboxylic acid

[0181]

[0182] 1.11 g of methyl 3-bromo-1H-indazole-5-carboxylic acid (4.35 mmol) was dissolved in 30 mL of DMF, and 574 μL of 1,1-difluoro-2-iodoethane (6.52 mmol) and 2.84 g of cesium carbonate (8.70 mmol) were added. The mixture was heated at 80 °C for 12 h. The reaction was monitored by TLC. After the reaction was completed, the mixture was quenched with a large amount of water and extracted three times with ethyl acetate (30 mL × 3). The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate for 30 min, and purified by column chromatography (petroleum ether:ethyl acetate = 16:1) to give 876 mg of white flocculent solid, with a yield of 63%.

[0183] 1 H NMR (300MHz, DMSO-d6) δ (ppm) 8.20 (dd, J=1.6, 0.7Hz, 1H), 8.08 (dd, J=8.9, 1.6Hz, 1H), 7.9 1(d,J=9.0Hz,1H),6.47(tt,J=54.4,3.4Hz,1H),5.03(td,J=15.5,3.4Hz,2H),3.90(s,3H).

[0184] Step 3. Synthesis of methyl 3-(5-chlorothiophene-2-yl)-1-(2,2-difluoroethyl)-1H-indazole-5-carboxylic acid

[0185]

[0186] 479 mg of methyl 3-bromo-1H-indazole-5-carboxylic acid (1.50 mmol) and 365 mg of 5-chlorothiophene-2-boric acid (2.25 mmol) were weighed and placed in a 100 mL two-necked round-bottom flask. A mixture of 16 mL toluene and 8 mL methanol was added to dissolve the solids. Then, 4 mL of an aqueous solution of 636 mg sodium carbonate (6.00 mmol) and 110 mg of [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (0.15 mmol) were added. The mixture was heated at 110 °C for 24 h under nitrogen protection. The reaction was monitored by TLC. After the reaction was complete, the mixture was filtered through diatomaceous earth, and the organic solvent was removed by vacuum distillation. The mixture was extracted three times with ethyl acetate (30 mL × 3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate for 30 min, and purified by column chromatography (petroleum ether:ethyl acetate = 16:1) to obtain 315 mg of a white powdery solid, with a yield of 59%.

[0187] 1 H NMR(300MHz,DMSO-d6)δ(ppm)8.64(dd,J=1.5,0.8Hz,1H),8.07(dd,J=8.9,1.5Hz,1H),7.90(d,J=8.9Hz,1H),7.7 1(d,J=4.0Hz,1H),7.28(d,J=3.9Hz,1H),6.49(tt,J=54.5,3.4Hz,1H),5.05(td,J=15.4,3.4Hz,2H),3.91(s,3H).

[0188] Step 4. Synthesis of 3-(5-chlorothiophene-2-yl)-1-(2,2-difluoroethyl)-1H-indazole-5-carboxylic acid

[0189]

[0190] 320 mg of methyl 3-(5-chlorothiophene-2-yl)-1-(2,2-difluoroethyl)-1H-indazole-5-carboxylic acid (0.93 mmol) was dissolved in 10 mL of methanol, and 2 mL of 209 mg of KOH (3.73 mmol) aqueous solution was added. After heating and refluxing at 70 °C for 2 hours, the reaction was monitored by TLC to be complete. The solvent was removed by vacuum distillation, and the product was dissolved in water. The pH was adjusted to acidic with 10% dilute hydrochloric acid under ice bath conditions, and a solid precipitated. After filtration and drying, 298 mg of white powder solid was obtained, with a yield of 97%.

[0191] 1H NMR (300MHz, DMSO-d6) δ (ppm) 13.04 (s, 1H), 8.63 (d, J = 1.4Hz, 1H), 8.06 (dd, J = 8.9, 1.4Hz, 1H), 7.88 (d, J = 8.9H z,1H),7.69(d,J=4.0Hz,1H),7.26(d,J=4.0Hz,1H),6.49(tt,J=54.5,3.4Hz,1H),5.04(td,J=15.4,3.4Hz,2H).

[0192] Step 5. Synthesis of 3-(5-chlorothiophene-2-yl)-N-cyclopropyl-1-(2,2-difluoroethyl)-1H-indazole-5-amide

[0193]

[0194] Using the product from step 4 (137 mg, 0.40 mmol) and cyclopropylamine (55 μL, 0.80 mmol) as raw materials, 96 mg of a white solid was obtained, with a yield of 60%.

[0195] MP200-202℃, 1 H NMR (300MHz, DMSO-d6) δ8.63 (d, J=3.9Hz, 1H), 8.51 (s, 1H), 7.99

[0196] (dd,J=9.0,1.5Hz,1H),7.83(d,J=8.9Hz,1H),7.79(d,J=4.0Hz,1H),7.31(d,J=3.9Hz,1H),6.47(tt,J=54.6,3 .5Hz,1H),5.01(td,J=15.4,3.5Hz,2H),2.87(tq,J=7.6,3.9Hz,1H),0.78–0.69(m,2H),0.65–0.58(m,2H)ppm; 13 C NMR (101MHz, DMSO) δ167.62,143.30,139.93,134.10,129.05,128.88,128.45,127.29,126.10,1 20.24,119.87,117.11,114.70,112.30,110.88,50.55(t,J=25.3Hz),23.59,6.32ppm.HRMS(ESI + forC 17 H 14 ClF2N3OS[M+H] + The measured value was 382.0585, and the theoretical value was 382.0584.

[0197] Example 2

[0198] Preparation of aziridine-1-yl(3-(5-chlorothiophen-2-yl)-1-(2,2-difluoroethyl)-1H-indazol-5-yl) methyl ketone

[0199]

[0200] Following the synthetic method described in 1, using aziridine hydrochloride (45 mg, 0.48 mmol) as the starting material, 96 mg of a white solid was obtained, with a yield of 63%.

[0201] MP108-110℃, 1 H NMR (300MHz, DMSO-d6) δ8.28 (s, 1H), 7.84 (d, J = 8.8Hz, 1H), 7.76

[0202] (dd,J=8.9,1.4Hz,1H),7.72(d,J=4.0Hz,1H),7.25(d,J=4.0Hz,1H),6.48(tt,J=54.6,3.4Hz,1H) ,5.03(td,J=15.4,3.4Hz,2H),4.38(t,J=7.7Hz,2H),4.09(t,J=7.6Hz,2H),2.34–2.20(m,2H)ppm; 13 C NMR (101MHz, CDCl3) δ169.95,142.57,140.85,133.38,130.68,127.43,127.24,126.91,124.69, 121.62, 120.80, 116.14, 113.71, 111.28, 109.38, 53.78, 51.16 (t, J = 28.6Hz), 49.10, 16.12ppm.

[0203] Example 3

[0204] Preparation of (3-(5-chlorothiophene-2-yl)-1-(2,2-difluoroethyl)-1H-indazol-5-yl)(3-fluorozahexacyclic butane-1-yl) methyl ketone

[0205]

[0206] Following the synthetic method described in 1, using 3-fluorozacriane hydrochloride (54 mg, 0.48 mmol) as the starting material, 96 mg of a white solid was obtained, with a yield of 60%.

[0207] MP124-126℃, 1H NMR (300MHz, DMSO-d6) δ8.30 (dd, J=1.4, 0.8Hz, 1H), 7.86 (d, J=

[0208] 8.8Hz,1H),7.78(dd,J=8.9,1.4Hz,1H),7.75(d,J=4.0Hz,1H),7.25(d,J=4.0Hz,1H),6.48(tt,J=54. 6,3.4Hz,1H),5.46(dtt,J=57.6,6.2,3.3Hz,1H),5.03(td,J=15.4,3.5Hz,2H),4.76–4.00(m,4H)ppm; 13 C NMR (101MHz, CDCl3) δ170.38 (d, J = 2.3Hz), 142.75, 140.96, 133.19, 130.86, 127.08, 126.96, 126.77, 124.7 9,121.89,120.88,116.11,113.68,111.24,109.60,83.25,81.22,51.19(t,J=28.6Hz)ppm.HRMS(ESI)forC 17 H 13 ClF3N3OS[M+H] + The theoretical value is 400.0490, and the measured value is 400.0498.

[0209] Example 4

[0210] Preparation of (3-(5-chlorothiophen-2-yl)-1-(2,2-difluoroethyl)-1H-indazol-5-yl)(3,3-difluoroazacyclobutane-1-yl) methyl ketone

[0211]

[0212] Following the synthesis method described in 1, using 3,3-difluoroazacyclobutane hydrochloride as a raw material, 60 mg of a white powdery solid was obtained, with a yield of 48%.

[0213] MP119-121℃, 1 H NMR(300MHz,DMSO-d6)δ8.34(t,J=1.2Hz,1H),7.88(d,J=9.0Hz,

[0214] 1H),7.82(dd,J=8.9,1.4Hz,1H),7.79(d,J=4.0Hz,1H),7.26(d,J=4.0Hz,1H),6.48(tt,J =54.5,3.4Hz,1H),5.04(td,J=15.5,3.4Hz,2H),4.70(d,J=80.4Hz,4H)ppm.HRMS(ESI)for C 17 H 12 ClF4N3OS[M+H] + :calcd,418.0396; found,418.0401.

[0215] Example 5

[0216] Preparation of (3-(5-chlorothiophen-2-yl)-1-(2,2-difluoroethyl)-1H-indazol-5-yl)(morpholino) methyl ketone

[0217]

[0218] Following the synthesis method described in 1, using morpholine as the starting material, 113 mg of a white solid was obtained, with a yield of 69%.

[0219] MP141-142℃, 1 H NMR (300MHz, CDCl3) δ8.06 (s, 1H), 7.53 (dd, J = 8.7, 1.4Hz, 1H),

[0220] 7.49(d,J=8.7Hz,1H),7.39(d,J=3.9Hz,1H),6.98(d,J=3.9Hz,1H),6.19(tt,J=55.4,4.3Hz,1H),4.72(td,J=13.4,4.3Hz,2H),3.72(s,8H)ppm; 13 C NMR (101MHz, CDCl3) δ170.15,142.07,140.61,133.33,130.73,129.31,126.91,126.69,124.69, 120.91,120.86,116.15,113.72,111.29,109.71,66.89,51.18(t,J=28.5Hz)ppm.HRMS(ESI)forC 18 H 16 ClF₂N₃O₂S[M+H] + :calcd,412.0686; found,412.0690.

[0221] Example 6

[0222] Preparation of (3,3-difluoroazacyclobutane-1-yl)(1-(2,2-difluoroethyl)-3-(5-methylthiophen-2-yl)-1H-indazol-5-yl) methyl ketone

[0223] Step 1. Preparation of methyl 1-(2,2-difluoroethyl)-3-(5-methylthiophen-2-yl)-1H-indazole-5-carboxylic acid

[0224]

[0225] Following the synthesis method in step 3 of Example 1, using (5-methylthiophen-2-yl)boronic acid as the raw material, 253 mg of a white solid was obtained, with a yield of 60%.

[0226] 1 H NMR (300MHz, DMSO-d6) δ8.64(dd,J=1.5,0.8Hz,1H),8.05(dd,J=8.9,1.5Hz,1H),7.87(d,J=8.9Hz,1H),7.58(d,J=3.6Hz, 1H), 6.95 (dd, J=3.6, 1.2Hz, 1H), 6.48 (tt, J=54.6, 3.4Hz, 1H), 5.02 (td, J=15.4, 3.4Hz, 2H), 3.90 (s, 3H), 2.52 (s, 3H)ppm.

[0227] Step 2. Preparation of 1-(2,2-difluoroethyl)-3-(5-methylthiophen-2-yl)-1H-indazole-5-carboxylic acid

[0228]

[0229] Following the synthesis method in step 4 of Example 1, 218 mg of a white solid was obtained, with a yield of 90%.

[0230] 1 H NMR (300MHz, DMSO-d6) δ13.05(s,1H),8.63(d,J=1.4Hz,1H),8.04(dd,J=8.8,1.5Hz,1H),7.84(d,J=8.9Hz,1H),7.57(d ,J=3.6Hz,1H),6.94(dd,J=3.6,1.3Hz,1H),6.48(tt,J=54.6,3.4Hz,1H),5.02(td,J=15.4,3.5Hz,2H),2.52(s,3H)ppm.

[0231] Step 3. Preparation of (3,3-difluoroazacyclobutane-1-yl)(1-(2,2-difluoroethyl)-3-(5-methylthiophene-2-yl)-1H-indazol-5-yl) methyl ketone

[0232]

[0233] Following the synthesis method in step 5 of Example 1, using 3,3-difluoroazacyclobutane hydrochloride as the starting material, 92 mg of a white solid was obtained, with a yield of 74%.

[0234] MP106-107℃, 1 H NMR(400MHz, DMSO-d6)δ8.33(t,J=1.1Hz,1H),7.84(d,J=8.9Hz,

[0235] 1H),7.80(dd,J=8.8,1.5Hz,1H),7.66(d,J=3.5Hz,1H),6.93(dd,J=3.6,1.3Hz,1H),6.47(t t,J=54.7,3.5Hz,1H),5.01(td,J=15.3,3.5Hz,2H),4.69(d,J=85.2Hz,4H),2.52(s,3H)ppm. 13 C NMR (101MHz, CDCl3) δ170.96–170.74(m),142.88,142.16,141.12,131.74,126.93,126.17,125.76 ,125.71,122.53,121.14,116.21,115.39,113.78,111.35,109.63,51.14(t,J=28.8Hz),15.42ppm.

[0236] Example 7

[0237] Preparation of (1-(2,2-difluoroethyl)-3-(5-methylthiophen-2-yl)-1H-indazol-5-yl)(morpholinyl) methyl ketone

[0238]

[0239] Following the synthesis method described in section 6, using morpholine as the starting material, 101 mg of a white solid was obtained, with a yield of 82%.

[0240] MP161-162℃, 1 H NMR (400MHz, CDCl3) δ8.11 (t, J = 1.1 Hz, 1H), 7.52 (dd, J = 8.7, 1.4

[0241] Hz,1H),7.47(d,J=8.6Hz,1H),7.43(d,J=3.5Hz,1H),6.83(dd,J=3.5,1.3Hz,1 H),6.19(tt,J=55.5,4.4Hz,1H),4.72(td,J=13.3,4.4Hz,2H),3.72(s,8H)ppm; 13 C NMR (101MHz, CDCl3) δ170.40,142.07,141.72,140.83,132.04,128.79,126.59,126.08,125.59 ,121.27,121.10,116.30,113.87,109.55,66.91,51.12(t,J=28.6Hz),15.39ppm.HRMS(ESI)for C 19 H 19 F2N3O2S[M+H] + :calcd,392.1236; found,392.1237.

[0242] Example 8

[0243] Preparation of (3-(2-chloropyridin-4-yl)-1-(2,2-difluoroethyl)-1H-indazol-5-yl)(3,3-difluoroazacyclobutane-1-yl) methyl ketone

[0244] Step 1. Preparation of methyl 3-(2-chloropyridin-4-yl)-1-(2,2-difluoroethyl)-1H-indazole-5-carboxylic acid

[0245]

[0246] Following the synthesis method in step 3 of Example 1, using 2-chloro-4-pyridineboronic acid (306 mg, 1.95 mmol, 1.50 eq) as the starting material, 252 mg of a white solid was obtained, with a yield of 55%.

[0247] 1 H NMR (300MHz, DMSO-d6) δ8.65(dd,J=1.5,0.8Hz,1H),8.56(dd,J=5.0,0.8Hz,1H),8.06(dd,J=9.0,1.5Hz,1H),7.97(dd ,J=5.1,1.5Hz,1H),7.94(d,J=9.1Hz,2H),6.52(tt,J=54.4,3.4Hz,1H),5.10(td,J=15.5,3.4Hz,2H),3.88(s,3H)ppm.

[0248] Step 2. Preparation of 3-(2-chloropyridin-4-yl)-1-(2,2-difluoroethyl)-1H-indazole-5-carboxylic acid

[0249]

[0250] Following the synthesis method in step 4 of Example 1, 213 mg of a white solid was obtained, with a yield of 90%.

[0251] 1 H NMR (300MHz, DMSO-d6) δ13.15(s,1H),8.70(t,J=1.1Hz,1H),8.61(dd,J=5.1,0.8Hz,1H),8.10(dd,J=8.9,1.4Hz,1H ),8.02(dd,J=6.6,1.4Hz,2H),7.97(d,J=8.9Hz,1H),6.57(tt,J=54.4,3.5Hz,1H),5.15(td,J=15.5,3.4Hz,2H)ppm.

[0252] Step 3. Preparation of (3-(2-chloropyridin-4-yl)-1-(2,2-difluoroethyl)-1H-indazol-5-yl)(3,3-difluoroazacyclobutane-1-yl) methyl ketone

[0253]

[0254] Following the synthesis method in step 5 of Example 1, using 3,3-difluoroazacyclobutane hydrochloride as the starting material, 92 mg of a white solid was obtained, with a yield of 74%.

[0255] MP154-155℃, 1 H NMR (300MHz, DMSO-d6) δ8.58 (dd, J=5.1, 0.7Hz, 1H), 8.43 (d, J=

[0256] 1.1Hz,1H),8.05(dd,J=5.2,1.5Hz,1H),8.02(dd,J=1.6,0.7Hz,1H),7.95(d,J=8.9Hz,1H),7.85(dd,J= 8.9,1.4Hz,1H),6.55(tt,J=54.4,3.4Hz,1H),5.14(td,J=15.5,3.4Hz,2H),4.69(d,J=82.0Hz,4H)ppm; 13C NMR (101MHz, CDCl3) δ170.66–170.54(m),152.46,150.39,143.17,142.72,142.67,127.08,126 .77,122.05,121.89,121.58,120.24,115.91,115.23,113.48,110.12,51.50(t,J=28.2Hz)ppm.

[0257] Example 9

[0258] Preparation of (3-(2-chloropyridin-4-yl)-1-(2,2-difluoroethyl)-1H-indazol-5-yl)(morpholino) methyl ketone

[0259]

[0260] Following the synthesis method in Example 8, using morpholine as a raw material, 101 mg of a white solid was obtained, with a yield of 82%.

[0261] MP188-191℃, 1 H NMR (300MHz, CDCl3) δ8.51 (d, J = 5.1Hz, 1H), 8.13 (d, J = 7.1Hz,

[0262] 1H),7.90(d,J=5.7Hz,1H),7.80(d,J=5.6Hz,1H),7.55(d,J=6.7Hz,2H),6 .22(tt,J=55.9,4.7Hz,1H),4.81(td,J=13.3,4.4Hz,2H),3.73(s,8H)ppm; 13 C NMR (101MHz, CDCl3) δ169.95,152.39,150.32,142.91,142.34,142.19,130.21,126.60,121. 83,121.55,120.64,120.21,113.54,110.07,66.85,51.46(t,J=28.2Hz)ppm.HRMS(ESI)forC 19 H 17 ClF2N4O2[M+H] + :calcd,407.1078; found,407.1094.

[0263] Example 10

[0264] Preparation of (3-(3-chloro-4-methoxyphenyl)-1-(2,2-difluoroethyl)-1H-indazol-5-yl)(3,3-difluoroazacyclobutane-1-yl) methyl ketone

[0265] Step 1. Preparation of methyl 3-(3-chloro-4-methoxyphenyl)-1-(2,2-difluoroethyl)-1H-indazole-5-carboxylic acid

[0266]

[0267] Following the synthesis method in step 3 of Example 1, using 3-chloro-4-methoxyphenylboronic acid as a raw material, 439 mg of a white powdery solid was obtained, with a yield of 82%.

[0268] 1 H NMR (300MHz, DMSO-d6) δ8.59(dd,J=1.5,0.7Hz,1H),8.05(dd,J=8.9,1.5Hz,1H),7.95–7.91(m,2H),7.91–7.87(m,1 H),7.37(d,J=8.4Hz,1H),6.53(tt,J=54.7,3.5Hz,1H),5.05(td,J=15.3,3.5Hz,2H),3.95(s,3H),3.90(s,3H)ppm.

[0269] Step 2. Preparation of 3-(3-chloro-4-methoxyphenyl)-1-(2,2-difluoroethyl)-1H-indazole-5-carboxylic acid

[0270]

[0271] Following the synthesis method in step 4 of Example 1, 432 mg of a white solid was obtained, with a yield of 98%.

[0272] 1 H NMR (300MHz, DMSO-d6) δ13.02(s,1H),8.63–8.55(m,1H),8.04(dd,J=8.9,1.4Hz,1H),7.95(d,J=2.0Hz,1H),7.92(dd,J=8.3,2.2 Hz,1H),7.86(d,J=8.9Hz,1H),7.37(d,J=8.5Hz,1H),6.52(tt,J=54.7,3.5Hz,1H),5.04(td,J=15.3,3.5Hz,2H),3.94(s,3H)ppm.

[0273] Step 3. Preparation of (3-(3-chloro-4-methoxyphenyl)-1-(2,2-difluoroethyl)-1H-indazol-5-yl)(3,3-difluoroazacyclobutane-1-yl) methyl ketone

[0274]

[0275] Following the synthesis method in step 5 of Example 1, using 3,3-difluoroazacyclobutane hydrochloride as the starting material, 52 mg of a white solid was obtained, with a yield of 29%.

[0276] MP126-127℃, 1 H NMR (300MHz, DMSO-d6) δ8.21 (s, 1H), 7.84 (dd, J = 10.7, 2.2Hz,

[0277] 2H),7.76(d,J=8.9Hz,1H),7.71(dd,J=8.9,1.4Hz,1H),7.20(d,J=8.4Hz,1H),6.44(tt ,J=54.7,3.5Hz,1H),4.95(td,J=15.2,3.5Hz,2H),4.86–4.29(m,4H),3.83(s,3H)ppm; 13 C NMR(101MHz, CDCl3)δ171.03–170.91(m),155.62,145.67,143.14,129.41,127.05,126.66,126.00,125 .70,123.38,122.74,121.65,116.29,115.45,113.86,112.43,109.69,56.41,51.22(t,J=28.4Hz)ppm.

[0278] Example 11

[0279] Preparation of (3-(3-chloro-4-methoxyphenyl)-1-(2,2-difluoroethyl)-1H-indazol-5-yl)(morpholinyl) methyl ketone

[0280]

[0281] Following the synthesis method of Example 10, using morpholine as a raw material, 108 mg of a white solid was obtained, with a yield of 62%.

[0282] MP202-203℃, 1 H NMR (300MHz, CDCl3) δ8.07 (s, 1H), 7.94 (d, J = 2.2Hz, 1H), 7.78 (dd,

[0283] J=8.5,2.2Hz,1H),7.50(s,2H),7.05(d,J=8.5Hz,1H),6.20(tt,J=55.5,4. 3Hz,1H),4.75(td,J=13.4,4.1Hz,2H),3.96(s,3H),3.89–3.43(m,8H)ppm;13 C NMR (101MHz, CDCl3) δ170.40,155.37,145.05,142.21,129.26,128.98,126.93,126.33,125.85,123.17,121 .49,121.25,116.27,113.84,112.29,111.41,109.54,66.89,56.30,51.20(t,J=28.4Hz)ppm.HRMS(ESI)for C 21 H 20 ClF2N3O3[M+H] + :calcd,436.1231; found,436.1235.

[0284] Example 12

[0285] Preparation of (3-(3,4-dichlorophenyl)-1-(2,2-difluoroethyl)-1H-indazol-5-yl)(morpholinyl) methyl ketone

[0286] Step 1. Preparation of methyl 3-(3,4-dichlorophenyl)-1-(2,2-difluoroethyl)-1H-indazole-5-carboxylic acid

[0287]

[0288] Following the synthesis method in step 3 of Example 1, using 3,4-dichlorophenylboronic acid (365 mg, 2.10 mmol, 1.30 eq) as the raw material, 412 mg of a white powdery solid was obtained, with a yield of 71%.

[0289] 1 H NMR(300MHz,DMSO-d6)δ8.61(dd,J=1.5,0.8Hz,1H),8.13(d,J=2.0Hz,1H),8.07(dd,J=8.9,1.5Hz,1H), 8.00–7.91(m,2H),7.85(d,J=8.4Hz,1H),6.54(tt,J=54.6,3.4Hz,1H),5.09(td,J=15.4,3.5Hz,2H)ppm.

[0290] Step 2. Preparation of 3-(3,4-dichlorophenyl)-1-(2,2-difluoroethyl)-1H-indazole-5-carboxylic acid

[0291]

[0292] Following the synthesis method in step 4 of Example 1, 417 mg of a white solid was obtained, with a yield of 94%.

[0293] 1 H NMR (300MHz, DMSO-d6) δ13.06(s,1H),8.61(d,J=1.3Hz,1H),8.14(d,J=2.0Hz,1H),8.06(dd,J=8.9,1.4Hz,1H),7.97(dd,J= 8.4, 2.1Hz, 1H), 7.91 (d, J = 8.9Hz, 1H), 7.84 (d, J = 8.4Hz, 1H), 6.54 (tt, J = 54.6, 3.4Hz, 1H), 5.08 (td, J = 15.3, 3.5Hz, 2H) ppm.

[0294] Step 3. (3-(3,4-dichlorophenyl)-1-(2,2-difluoroethyl)-1H-indazol-5-yl)(morpholino) methyl ketone

[0295]

[0296] Following the synthesis method in step 5 of Example 1, using morpholine (70 μL, 0.80 mmol) as the starting material, 120 mg of a white solid was obtained, with a yield of 72%.

[0297] MP154-156℃, 1 H NMR(300MHz,DMSO-d6)δ8.13(d,J=2.1Hz,1H),8.12(s,1H),7.98

[0298] (dd,J=8.4,2.1Hz,1H),7.87(d,J=8.7Hz,1H),7.77(d,J=8.4Hz,1H),7.54(dd,J=8.7,1 .4Hz,1H),6.50(tt,J=54.6,3.4Hz,1H),5.05(td,J=15.3,3.5Hz,2H),3.58(s,8H)ppm; 13 C NMR (101MHz, CDCl3) δ170.21,144.06,142.26,133.27,132.74,132.42,130.97,129.49,129.15, 126.57,126.46,121.45,120.98,113.71,109.73,66.88,51.30(t,J=28.3Hz)ppm.HRMS(ESI)for C 20 H 17 Cl2F2N3O2[M+H] + :calcd,440.0736; found,440.0743.

[0299] Example 13

[0300] Preparation of 1-(4-(3-(3,4-dichlorophenyl)-1-(2,2-difluoroethyl)-1H-indazole-5-carbonyl)piperazin-1-yl)ethyl-1-one

[0301]

[0302] Following the synthesis method described in Example 12, using N-acetylpiperazine (71 μL, 0.60 mmol) as the starting material, 72 mg of a white solid was obtained, with a yield of 50%.

[0303] MP144-145℃, 1 H NMR (300MHz, CDCl3) δ8.09 (d, J = 1.2 Hz, 1H), 8.03 (d, J = 2.0 Hz,

[0304] 1H),7.77(dd,J=8.3,2.0Hz,1H),7.58(d,J=8.3Hz,1H),7.53(d,J=1.5Hz,2H),6.22(tt ,J=55.3,4.2Hz,1H),4.78(td,J=13.4,4.2Hz,2H),3.84–3.50(m,8H),2.14(s,3H)ppm; 13 C NMR (101MHz, DMSO) δ169.68,168.92,142.55,142.52,133.35,132.31,131.78,131.40,130.30,128.79,127 .61,126.73,120.51,117.23,114.83,112.43,111.30,50.69(t,J=25.3Hz),45.97,21.73ppm.HRMS(ESI)for C 22 H 20 Cl2F2N4O2[M+H] + :calcd,481.1001; found,481.1013.

[0305] Example 14

[0306] Preparation of 1-(3-(3,4-dichlorophenyl)-1-(2,2-difluoroethyl)-1H-indazole-5-carbonyl)pyrrolidine-3-one

[0307]

[0308] Following the synthesis method of Example 12, using 3-pyrrolidone hydrochloride as a starting material, 102 mg of a white solid was obtained, with a yield of 58%.

[0309] MP117-119℃, 1 H NMR (300MHz, CDCl3) δ8.16 (s, 1H), 8.03 (d, J = 2.0Hz, 1H), 7.77 (dd,

[0310] J=8.3,2.1Hz,1H),7.64(dd,J=8.8,1.4Hz,1H),7.58(d,J=8.4Hz,1H),7.54(d,J=8.9Hz,1H),6. 22(tt,J=55.3,4.2Hz,1H),4.78(td,J=13.4,4.2Hz,2H),4.07(s,4H),2.66(t,J=7.8Hz,2H)ppm; 13 C NMR(101MHz,DMSO)δ211.58,169.35,142.70,142.67,133.33,132.31,131.77,131.42,128.81,1 27.62,126.80,120.42,117.23,114.82,112.42,111.24,50.69(t,J=25.2Hz)ppm.HRMS(ESI)for C 20 H 15 Cl2F2N3O2[M+H] + :calcd,438.0579; found,438.0592.

[0311] Example 15

[0312] Preparation of (3-(3,4-dichlorophenyl)-1-(2,2-difluoroethyl)-1H-indazol-5-yl)(3,3-difluoroazacyclobutane-1-yl) methyl ketone

[0313]

[0314] Following the synthesis method of Example 12, using 3,3-difluoroazacyclobutane hydrochloride (57 mg, 0.44 mmol) as the starting material, 76 mg of a white solid was obtained, with a yield of 43%.

[0315] MP137-138℃, 1 H NMR(300MHz,DMSO-d6)δ8.25(d,J=1.4Hz,1H),8.05(d,J=2.0

[0316] Hz,1H),7.91(dd,J=8.4,2.1Hz,1H),7.83(d,J=8.8Hz,1H),7.76(dd,J=8.9,1.5Hz,1H),7.72(d,J= 8.4Hz, 1H), 6.47 (tt, J = 54.6, 3.4Hz, 1H), 5.01 (td, J = 15.4, 3.4Hz, 2H), 4.63 (d, J = 69.9Hz, 4H) ppm; 13 C NMR (101MHz, CDCl3) δ170.95–170.68(m),144.58,143.08,133.36,132.96,132.17,131.03,129.20, 126.65,126.59,126.38,122.38,121.49,116.07,115.29,113.64,109.79,51.33(t,J=28.4Hz)ppm.

[0317] Example 16

[0318] Preparation of (3-(3,4-dichlorophenyl)-1-(2,2-difluoroethyl)-1H-indazol-5-yl)(4-methylpiperazin-1-yl) methyl ketone

[0319]

[0320] Following the synthesis method of Example 12, using N-methylpiperazine as a starting material, 99 mg of a white solid was obtained, with a yield of 55%.

[0321] MP177-179℃, 1 H NMR (300MHz, CDCl3) δ8.07 (t, J = 1.1 Hz, 1H), 8.04 (d, J = 2.0 Hz,

[0322] 1H),7.78(dd,J=8.4,2.0Hz,1H),7.58(d,J=8.3Hz,1H),7.52(d,J=1.3Hz,2H),6.21(tt,J=55 .3,4.2Hz,1H),4.77(td,J=13.4,4.3Hz,2H),3.96–3.51(m,4H),2.44(s,4H),2.33(s,3H)ppm; 13C NMR(101MHz,DMSO)δ169.39,142.47,133.36,132.29,131.76,131.36,130.55,128.76,127.58,1 26.64,120.52,120.30,117.22,114.82,112.41,111.24,54.93,50.69(t,J=25.3Hz),46.08ppm.

[0323] Example 17

[0324] Preparation of (3-(3,4-dichlorophenyl)-1-(2,2-difluoroethyl)-1H-indazol-5-yl)(4-hydroxypiperidin-1-yl) methyl ketone

[0325]

[0326] Following the synthesis method of Example 12, using 4-hydroxypiperidine as a starting material, 89 mg of a white solid was obtained, with a yield of 49%.

[0327] MP106-107℃, 1 H NMR (300MHz, CDCl3) δ8.06 (d, J = 1.2 Hz, 1H), 8.04 (d, J = 2.0 Hz,

[0328] 1H),7.78(dd,J=8.3,2.0Hz,1H),7.58(d,J=8.3Hz,1H),7.52(d,J=1.1Hz,2H),6.22(tt,J=55.3,4.2Hz,1H),4.77(td,J=1 3.4, 4.3Hz, 2H), 4.01 (tt, J = 7.2, 3.4Hz, 1H), 3.36 (s, 2H), 1.89 (d, J = 24.0Hz, 2H), 1.62 (s, 4H), 0.86 (d, J = 6.7Hz, 1H) ppm.

[0329] Example 18

[0330] Preparation of (3-(3,4-dichlorophenyl)-1-(2,2-difluoroethyl)-1H-indazol-5-yl)(2-oxa-6-azaspiro[3.3]hept-6-yl) methyl ketone

[0331]

[0332] Following the synthesis method of Example 12, using 2-oxa-6-aza-spiro[3,3]heptane oxalate as a starting material, 73 mg of a white solid was obtained, with a yield of 40%.

[0333] MP116-118℃,1 H NMR (300MHz, CDCl3) δ8.27 (d, J = 1.3 Hz, 1H), 8.04 (d, J = 2.0 Hz,

[0334] 1H),7.78(dd,J=8.3,2.0Hz,1H),7.73(dd,J=8.8,1.5Hz,1H),7.59(d,J=8.3Hz,1H),7 .51(d,J=8.8Hz,1H),6.22(tt,J=55.3,4.2Hz,1H),4.89–4.70(m,6H),4.44(s,4H)ppm; 13 C NMR (101MHz, DMSO) δ169.42,143.16,142.91,133.24,132.36,131.90,131.51,128.86,127.78,127.62,126.93,12 1.33,120.57,117.22,114.81,112.41,111.18,80.16,62.63,58.44,50.69(t,J=25.4Hz),38.17ppm.HRMS(ESI)for C 21 H 17 Cl2F2N3O2[M+H] + :calcd,452.0736; found,452.0742.

[0335] Example 19

[0336] Preparation of (3-(3,4-dichlorophenyl)-1-(2,2-difluoroethyl)-1H-indazol-5-yl)(2-oxa-7-azaspiro[3.5]non-7-yl) methyl ketone

[0337]

[0338] 100 mg (0.44 mmol) of tert-butyl 2-oxa-7-diazaspiro[3.5]nonane-7-carboxylate was dissolved in 5 mL of dichloromethane, and 0.5 mL of dioxane hydrochloride solution was added. After reacting at room temperature for 2 hours, TLC showed that the reaction was complete, and saturated sodium bicarbonate solution was added until no more bubbles were produced. The pH was adjusted to 9-10 with sodium hydroxide solution, and the product was extracted three times with dichloromethane (5 mL × 3). After drying with anhydrous sodium sulfate for 30 min, the product was evaporated to dryness to obtain an oily de-Boc product, which was directly used in the next reaction. Following the synthesis method in Example 12, 100 mg of a white solid was obtained, with a yield of 52%.

[0339] MP120-121℃, 1H NMR (300MHz, CDCl3) δ8.06 (t, J = 1.1 Hz, 1H), 8.04 (d, J = 2.0 Hz,

[0340] 1H),7.78(dd,J=8.4,2.0Hz,1H),7.58(d,J=8.3Hz,1H),7.52(d,J=1.1Hz,2H),6.22(tt ,J=55.4,4.3Hz,1H),4.77(td,J=13.4,4.2Hz,2H),3.81–3.32(m,8H),1.65(s,4H)ppm; 13 C NMR (101MHz, DMSO) δ169.37,142.45,133.39,132.31,131.78,131.37,130.89,128.79,127.58,126. 54,120.53,120.00,117.23,114.82,112.42,111.25,62.73,50.69(t,J=25.6Hz),50.02,38.64ppm.

[0341] Example 20

[0342] Preparation of (3-(3-chlorophenyl)-1-(2,2-difluoroethyl)-1H-indazol-5-yl)(3,3-difluoroazacyclobutane-1-yl) methyl ketone

[0343] Step 1. Preparation of methyl 3-(3-chlorophenyl)-1-(2,2-difluoroethyl)-1H-indazole-5-carboxylic acid

[0344]

[0345] Following the synthesis method in step 3 of Example 1, using 3-chlorophenylboronic acid as a raw material, 450 mg of a white powdery solid was obtained, with a yield of 91%.

[0346] 1 H NMR (300MHz, DMSO-d6) δ8.62(dd,J=1.5,0.8Hz,1H),8.07(dd,J=8.9,1.5Hz,1H),7.97–7.94(m,2H),7.94–7.91(m,1H),7.64(t ,J=8.0Hz,1H),7.56(ddd,J=8.1,2.1,1.3Hz,1H),6.54(tt,J=54.5,3.4Hz,1H),5.09(td,J=15.4,3.5Hz,2H),3.90(s,3H)ppm.

[0347] Step 2. Preparation of 3-(3-chlorophenyl)-1-(2,2-difluoroethyl)-1H-indazole-5-carboxylic acid

[0348]

[0349] Following the synthesis method in step 4 of Example 1, 317 mg of a white solid was obtained, with a yield of 94%.

[0350] 1 H NMR (300MHz, DMSO-d6) δ13.08(s,1H),8.65–8.60(m,1H),8.07(dd,J=8.9,1.4Hz,1H),7.97(d,J=1.9Hz,1H),7.95(dd,J=3.0,1.6Hz,1H), 7.92(d,J=8.9Hz,1H),7.64(t,J=8.0Hz,1H),7.57(dt,J=8.2,1.4Hz,1H),6.55(tt,J=54.6,3.5Hz,1H),5.10(td,J=15.4,3.5Hz,2H)ppm.

[0351] Step 3. Preparation of (3-(3-chlorophenyl)-1-(2,2-difluoroethyl)-1H-indazol-5-yl)(3,3-difluoroazacyclobutane-1-yl) methyl ketone

[0352]

[0353] Following the synthesis method in step 5 of Example 1, using 3,3-difluoroazacyclobutane hydrochloride as the starting material, 75 mg of a white solid was obtained, with a yield of 61%.

[0354] MP104-106℃, 1 H NMR (300MHz, DMSO-d6) δ8.34 (dd, J=1.5, 0.8Hz, 1H), 7.97 (d, J=

[0355] 1.6Hz, 2H), 7.90 (d, J=8.8Hz, 1H), 7.82 (dd, J=8.9, 1.5Hz, 1H), 7.61 (dd, J=7.8, 1.6Hz, 1H), 7.55 (dt, J= 8.1,1.4Hz,1H),6.53(tt,J=54.6,3.4Hz,1H),5.09(td,J=15.4,3.5Hz,2H),4.69(d,J=64.6Hz,4H)ppm; 13C NMR (101MHz, CDCl3) δ170.91 (t, J = 3.3Hz), 145.60, 143.07, 135.07, 133.90, 130.35, 129.10, 127.58, 126 .66,126.20,125.66,122.52,121.61,116.14,115.32,113.71,111.28,109.73,51.30(t,J=28.4Hz)ppm.

[0356] Example 21

[0357] Preparation of (3-(3-chlorophenyl)-1-(2,2-difluoroethyl)-1H-indazol-5-yl)(morpholinyl) methyl ketone

[0358]

[0359] Following the synthesis method of Example 20, using morpholine as a raw material, 63 mg of a white solid was obtained, with a yield of 52%.

[0360] MP185-186℃, 1 H NMR (400MHz, CDCl3) δ8.11 (t, J = 1.2 Hz, 1H), 7.92 (t, J = 1.8 Hz,

[0361] 1H),7.81(dt,J=7.4,1.6Hz,1H),7.53(d,J=1.2Hz,2H),7.46(t,J=7.7Hz,1H),7.41(dt,J=8 .1,1.7Hz,1H),6.22(tt,J=55.4,4.3Hz,1H),4.78(td,J=13.4,4.3Hz,2H),3.73(s,8H)ppm.

[0362] Example 22

[0363] Preparation of (3-(3-chlorophenyl)-1-(2,2-difluoroethyl)-1H-indazol-5-yl)(8-oxo-2-azaspiro[4.5]dec-2-yl) methyl ketone

[0364]

[0365] Following the synthesis method of Example 20, 53 mg of a white powdery solid product was obtained from 8-oxa-2-azaspiro[4.5]decane, with a yield of 65%.

[0366] 1H NMR (300MHz, CDCl3) δ8.18(s,1H),7.92(s,1H),7.82(d,J=6.8Hz,1H),7.64(dd,J=8.7 ,1.4Hz,1H),7.55–7.38(m,3H),6.23(td,J=55.3,4.0Hz,1H),4.78(td,J=14.0,4.4Hz ,2H),3.83–3.74(m,2H),3.67(d,J=19.1Hz,3H),3.60–3.49(m,2H),3.37(s,1H),1.93 (t,J=7.3Hz,1H),1.82(t,J=7.0Hz,1H),1.67(t,J=5.6Hz,2H),1.55(d,J=5.0Hz,2H). 13 C NMR (101MHz, CDCl3) δ145.16,142.30,134.95,134.23,130.27,128.68,127.55,126.48,125.68,121.41,120.95,116.2 3,113.80,111.37,109.46,65.19,64.96,51.27(t,J=28.3Hz),47.86,44.63,40.70,38.62,35.50,34.69ppm.HRMS(ESI + ):m / z[M+H + Calculated for C 24 H 24 ClF2N3O2,460.16034; found460.16213.

[0367] Example 23

[0368] Preparation of 7-(3-(3-chlorophenyl)-1-(2,2-difluoroethyl)-1H-indazole-5-carbonyl)-7-azaspiro[3.5]non-2-one

[0369]

[0370] Following the synthesis method of Example 20, using 7-azaspiro[3.5]nonane-2-one hydrochloride as the raw material, 89 mg of a white powdery solid product was obtained, with a yield of 67%.

[0371] 1H NMR (300MHz, CDCl3) δ8.09 (s, 1H), 7.92 (d, J = 2.0Hz, 1H), 7.81 (dt, J = 7.2, 1.7Hz, 1H), 7.53 (d, J = 1.1Hz, 2H), 7.49 -7.37(m,2H),6.22(tt,J=55.4,4.2Hz,1H),4.78(td,J=13.4,4.3Hz,2H),3.63(d,J=38.0Hz,4H),2.88(s,4H),1.81(s,4H)ppm. 13 C NMR (101MHz, CDCl3) δ205.91,170.40,145.00,142.21,134.94,134.20,130.29,129.86,128.69,127.51,126.33, 125.64,121.52,120.80,116.23,113.80,111.37,109.65,56.51,51.26(t,J=28.3Hz),29.44,26.91ppm.HRMS(ESI + ):m / z[M+H + Calculated for C 24 H 22 ClF2N3O2,458.1447; found 458.1446.

[0372] Example 24

[0373] Preparation of (3-(3-chlorophenyl)-1-(2,2-difluoroethyl)-1H-indazol-5-yl)(9-methyl-3,9-diazaspiro[5.5]undecane-3-yl) methyl ketone

[0374]

[0375] Following the synthesis method of Example 20, using 3-methyl-3,9-diazaspiro[5,5]undecane (61 mg, 0.36 mmol, 1.20 eq) as the starting material, 92 mg of a white powdery solid product was obtained, with a yield of 64%.

[0376] 1H NMR (300MHz, CDCl3) δ8.07(s,1H),7.91(s,1H),7.81(d,J=6.9Hz,1H),7.55(s,2H),7.43(d,J=7.7Hz,2H),6.22(t,J=25.4H z,1H),4.77(dt,J=13.4,7.9Hz,2H),3.59(d,J=29.1Hz,4H),2.70(s,4H),2.53(s,3H),1.79(s,4H),1.72–1.43(m,4H)ppm. 13 C NMR (101MHz, CDCl3) δ170.15,144.96,142.12,134.92,134.26,130.27,130.24,128.62,127.50,126.40,125.64, 121.49,120.69,116.24,113.81,111.38,109.47,51.25(t,J=29.3Hz),51.14,46.39,35.57,29.53ppm.HRMS(ESI + ):m / z[M+H + Calculated for C 26 H 29 ClF2N4O,487.2076; found 487.2060.

[0377] Example 25

[0378] Preparation of (3-(3-chlorophenyl)-1-(2,2-difluoroethyl)-1H-indazol-5-yl)(6-methoxy-2-azaspiro[3.3]heptane-2-yl) methyl ketone

[0379]

[0380] The starting material, tert-butyl 6-hydroxy-2-azaspiro[3.3]heptane-2-carboxylate (250 mg, 1.17 mmol, 1.00 eq), was dissolved in 6 mL of anhydrous N,N-dimethylformamide. NaH (56 mg, 2.34 mmol, 2.00 eq) was slowly added under ice bath (0 °C), followed by stirring at room temperature for 30 min. Iodomethane (109 μL, 1.76 mmol, 1.50 eq) was then added, and the reaction continued at room temperature for 1 h. After TLC analysis, a large amount of water was added to quench the reaction mixture, followed by extraction three times with ethyl acetate (20 mL × 3). The combined organic phases were washed twice with saturated brine, dried over anhydrous sodium sulfate for 0.5 h, and concentrated under reduced pressure to obtain a colorless oily product, methyl ether, which was directly used in the next reaction. The methyl ether was dissolved in 2 mL of ethyl acetate, followed by the addition of 1 mL of dioxane hydrochloride solution. The mixture was stirred at room temperature for 3 h, and then evaporated to dryness to obtain a white de-Boc solid product, which was directly used in the next reaction. Following the synthesis method of Example 20, 63 mg of a yellowish-white flaky solid product was obtained, with a yield of 48%.

[0381] 1 H NMR (300MHz, CDCl3) δ8.30(s,1H),7.93(s,1H),7.82(dt,J=7.3,1.7Hz,1H),7.75(dd,J=8.8,1.5Hz,1H),7.52–7.40(m,3H),6.22(tt,J=5 5.5,4.3Hz,1H),4.77(td,J=13.4,4.3Hz,2H),4.26(d,J=15.1Hz,4H),3.78(d,J=28.9Hz,1H),3.22(s,3H),2.55(s,2H),2.14(s,2H)ppm. 13 C NMR (101MHz, CDCl3) δ170.08,145.35,142.77,134.95,134.13,130.28,128.72,127.53,126.86,125.66,122 .07,121.47,116.19,113.76,111.33,109.33,69.57,55.47,51.24(t,J=28.3Hz),40.80,38.61ppm.HRMS(ESI + ):m / z[M+H + calculated for C 23 H 22 ClF2N3O2,446.14469; found446.14389.

[0382] Example 26

[0383] Preparation of (3-(3-chlorophenyl)-1-(2,2-difluoroethyl)-1H-indazol-5-yl)(2-oxa-6-azaspiro[3.4]oct-6-yl) methyl ketone

[0384]

[0385] 60 mg (0.28 mmol) of 2-oxa-6-azaspiro[3,4]octane-6-carboxylic acid tert-butyl ester was dissolved in 4 mL of dichloromethane, and 0.5 mL of trifluoroacetic acid solution was slowly added dropwise with stirring. The reaction was carried out at room temperature for 2 h. After the reaction was completed by TLC, the organic solvent was removed by vacuum distillation, an appropriate amount of water was added, and the pH was adjusted to 7-8 with saturated sodium bicarbonate solution. The product was extracted three times with ethyl acetate (15 mL × 3), and the organic phases were collected and combined. The product was washed twice with saturated brine, dried over anhydrous sodium sulfate for 0.5 h, and concentrated under reduced pressure to obtain a colorless oily product, which was directly used for the next reaction. Following the synthesis method of Example 20, 49 mg of a white powdery solid product was obtained, with a yield of 64%.

[0386] 1 H NMR (300MHz, CDCl3) δ8.18(s,1H),7.92(s,1H),7.81(s,1H),7.64(d,J=8.3Hz,1H),7.60–7.36(m,3H), 6.23(t,J=55.4Hz,1H),4.80(s,2H),4.77–4.44(m,4H),3.73(t,J=60.1Hz,4H),2.40–2.09(m,2H)ppm. 13 C NMR (101MHz, CDCl3) δ169.69,145.21,142.37,134.96,134.17,130.28,128.72,127.56,126.46,125.68,121.39 ,121.13,116.22,113.79,111.35,109.48,80.69,80.07,58.22,54.96,51.28(t,J=27.3Hz),48.57ppm.HRMS(ESI + ):m / z[M+H + Calculated for C 22 H 20 ClF2N3O2,432.12904; found 432.12861.

[0387] Example 27

[0388] Preparation of (4-(3-chlorophenyl)-1-(2,2-difluoroethyl)-1H-indazole-5-yl)(6-hydroxy-2-azaspiro[3.4]oct-2-yl) methyl ketone

[0389]

[0390] Following the synthesis method of Example 26, using 6-hydroxy-2-azaspiro[3.4]octane-2-carboxylic acid tert-butyl ester as the starting material, 78 mg of a white powder solid product was obtained, with a yield of 59%.

[0391] 1 H NMR(400MHz, CDCl3)δ8.33(s,1H),7.93(t,J=1.9Hz,1H),7.82(d,J=7.5Hz,1H), 7.76(dd,J=8.8,1.4Hz,1H),7.52–7.38(m,3H),6.22(tt,J=55.4,4.3Hz,1H),4.7 7(td,J=13.4,4.4Hz,2H),4.43–4.33(m,1H),4.21(d,J=21.9Hz,2H),4.13(s,1H) ,2.16(s,1H),2.08(dd,J=13.9,5.6Hz,1H),1.97(d,J=13.4Hz,2H),1.71(s,4H). 13 C NMR (101MHz, CDCl3) δ170.14,145.40,142.75,134.96,134.12,130.29,128.73,127.55,126.90,125.69,122.18,1 21.49,116.20,113.77,111.34,109.27,73.26,66.98,60.28,51.25(t,J=28.3Hz),40.77,36.61,34.85.HRMS(ESI + ):m / z[M+H + Calculated for C 23 H 22 ClF2N3O2,446.1447; found 446.1425.

[0392] Example 28

[0393] Preparation of (3-(3-chlorophenyl)-1-(2,2-difluoroethyl)-1H-indazol-5-yl)(piperazin-1-yl) methyl ketone

[0394]

[0395] The starting material (60 mg, 0.179 mmol, 1.00 eq) was dissolved in 6 mL of anhydrous dichloromethane. EDCI (51 mg, 0.27 mmol, 1.50 eq) and HOBT (36 mg, 0.27 mmol, 1.50 eq) were added sequentially. After stirring at room temperature for 0.5 h to activate the reaction, N-Boc-piperazine (40 mg, 0.21 mmol, 1.20 eq) and DIPEA (62 μL, 0.36 mmol, 2.00 eq) were added, and the reaction was continued at room temperature for 3 h. After the reaction was completed by TLC, 30 mL of water was added to the reaction solution, and the mixture was extracted three times with dichloromethane (20 mL × 3). The organic phases were collected and combined, and washed sequentially with 1 mol / L hydrochloric acid solution (20 mL × 2), saturated sodium bicarbonate solution (20 mL × 2), and saturated brine solution (20 mL × 2). The mixture was then dried over anhydrous sodium sulfate for 0.5 h. After concentrating the organic phase under reduced pressure, column chromatography (dichloromethane:methanol = 100:1) yielded a white powdery solid intermediate product, which was directly used for the next reaction. The intermediate was dissolved in 3 mL of dichloromethane, and then 1 mL of trifluoroacetic acid solution was added. The mixture was stirred at room temperature for 2 h. After the reaction was completed by TLC, the organic solvent was removed under reduced pressure, an appropriate amount of water was added, and the pH was adjusted to 7-8 with saturated sodium bicarbonate solution. The mixture was extracted three times with ethyl acetate (15 mL × 3), and the combined organic phases were collected, washed twice with saturated brine, dried over anhydrous sodium sulfate for 0.5 h, and concentrated under reduced pressure to obtain 47 mg of a white powdery solid product, with a yield of 65%.

[0396] 1 H NMR (300MHz, CDCl3) δ8.10(s,1H),7.92(t,J=1.9Hz,1H),7.81(dt,J=7.2,1.7Hz,1H),7.52(s,2H),7.49–7.34(m,2H ),6.22(tt,J=55.3,4.2Hz,1H),4.78(td,J=13.4,4.3Hz,2H),3.61(d,J=80.0Hz,4H),2.91(s,4H),1.81(s,1H)ppm. 13 C NMR (101MHz, CDCl3) δ170.27,145.03,142.16,134.97,134.23,130.24,129.90,128.67,127.52,12 6.47,125.63,121.55,120.95,116.23,113.80,111.37,109.53,51.28(t,J=28.3Hz)ppm.HRMS(ESI + ):m / z[M+H + Calculated for C 20 H 19ClF2N4O,405.1294; found 405.1288.

[0397] Example 29

[0398] Preparation of (3-(3-chlorophenyl)-1-(2,2-difluoroethyl)-1H-indazol-5-yl)(1,3-oxazin-3-yl) methyl ketone

[0399]

[0400] Following the synthesis method of Example 20, using 1,3-oxazine as a starting material, 67 mg of a white powdery solid product was obtained, with a yield of 92%.

[0401] 1 H NMR (300MHz, CDCl3) δ8.17(d,J=1.3Hz,1H),7.93(d,J=2.0Hz,1H),7.82(dt,J=7.2,1.7Hz,1H),7.63–7.50(m,2H),7.50– 7.38(m,2H),6.22(tt,J=55.4,4.3Hz,1H),4.95(s,2H),4.78(td,J=13.4,4.3Hz,2H),3.96(t,J=5.3Hz,4H),1.86(s,2H). 13 C NMR (101MHz, CDCl3) δ170.42,145.15,142.43,134.96,134.16,130.24,129.06,128.69,127.52,126. 70,125.66,121.50,116.21,113.78,111.35,109.56,68.26,51.27(t,J=29.3Hz),26.14ppm.HRMS(ESI + ):m / z[M+H + Calculated for C 20 H 18 ClF2N3O2,406.1134; found 406.1136.

[0402] Example 30

[0403] Preparation of (3-(3-chlorophenyl)-1-(2,2-difluoroethyl)-1H-indazol-5-yl)(1,4-oxacyclopent-4-yl) methyl ketone

[0404]

[0405] Following the synthesis method of Example 20, using 1,4-oxazacycloheptane as a raw material, a colorless oily product was obtained, yielding 67 mg of a white powdery solid with a yield of 90%.

[0406] 1 H NMR(300MHz, CDCl3)δ8.07(s,1H),7.92(t,J=1.9Hz,1H),7.81(dt,J=7.2,1.7Hz,1H),7.53(s,2H),7.49–7.38(m ,2H),6.22(tt,J=25.4,4.3Hz,1H),4.78(td,J=13.4,4.3Hz,2H),3.97–3.52(m,8H),1.97(d,J=32.2Hz,2H)ppm. 13 C NMR (101MHz, CDCl3) δ171.48,144.98,142.06,134.97,134.23,130.60,130.26,128.67,127.51,126.19,125. 63,121.46,120.25,116.23,113.80,111.37,109.66,71.15,69.59,69.21,51.27(t,J=28.3Hz)ppm.HRMS(ESI + ):m / z[M+H + Calculated for C 21 H 20 ClF2N3O2,420.1290; found420.1279.

[0407] Example 31

[0408] Preparation of 1-(3-(3-chlorophenyl)-1-(2,2-difluoroethyl)-1H-indazole-5-carbonyl)piperidin-4-one

[0409]

[0410] The starting material 1-(tert-butoxycarbonyl)-4-piperidinone (200 mg, 1.00 mmol) was dissolved in 4 mL of dichloromethane, and 2 mL of dioxane hydrochloride solution was added. The reaction was carried out at room temperature with stirring for 3 h. After the reaction was completed by TLC, the solvent was removed by vacuum distillation, an appropriate amount of water was added, and the pH was adjusted to 8-9 with saturated sodium bicarbonate solution. The mixture was extracted three times with dichloromethane, and the combined organic phases were washed twice with saturated brine. The mixture was dried over anhydrous sodium sulfate for 0.5 h and concentrated under reduced pressure to obtain 80 mg of a white powdery solid intermediate, which was directly added to the next reaction. Following the synthesis method of Example 20, 62 mg of a white powdery solid product was obtained, with a yield of 84%.

[0411] 1 H NMR(300MHz, CDCl3)δ8.17(s,1H),7.92(d,J=2.0Hz,1H),7.81(dt,J=7.1,1.7Hz,1H),7.62–7.53(m,2H), 7.49–7.40(m,2H),6.23(tt,J=55.4,4.2Hz,1H),4.79(td,J=13.4,4.3Hz,2H),3.94(s,4H),2.54(s,4H). 13 C NMR (101MHz, DMSO) δ207.82,170.10,143.47,142.58,134.83,134.23,131.51,130.27,128.72,126.86 ,126.49,126.21,120.62,120.36,117.26,114.85,112.45,111.25,50.70(t,J=25.3Hz)ppm.HRMS(ESI + ):m / z[M+H + Calculated for C 21 H 18 ClF2N3O2,418.11339; found 418.11369.

[0412] Example 32

[0413] Preparation of ((1R,4S)-2-oxa-5-azabicyclo[2.2.1]heptane-5-yl)(3-(3-chlorophenyl)-1-(2,2-difluoroethyl)-1H-indazol-5-yl) methyl ketone

[0414]

[0415] Following the synthesis method of Example 20, using bridged morpholine hydrochloride as a raw material, 63 mg of a white powdery solid product was obtained, with a yield of 84%.

[0416] 1H NMR (300MHz, CDCl3) δ8.19(s,1H),7.91(s,1H),7.79(s,1H),7.72–7.57(m,1H),7.53(d,J=8.3Hz,1H),7.46(d,J=8.4Hz,1H),7.42(s,1H),6.2 2(t,J=55.3Hz,1H),5.14–4.39(m,4H),4.20–3.97(m,1H),3.89(d,J=14.6Hz,1H),3.78–3.55(m,1H),3.49(s,1H),1.93(d,J=26.5Hz,2H)ppm. 13 C NMR (101MHz, CDCl3) δ134.98,134.13,130.30,128.73,127.54,126.48,125.67,121.31,116.22,113.78,1 11.35,109.47,75.99,74.25,60.54,58.42,56.86,54.55,51.27(t,J=28.3Hz),37.27,35.73ppm.HRMS(ESI + ):m / z[M+H + Calculated for C 21 H 18 ClF2N3O2,418.1134; found 418.1136.HPLC purity=96.56%,t R = 14.05 min.

[0417] Example 33

[0418] Preparation of (3,6-diazabicyclo[3.1.1]heptane-3-yl)(3-(3-chlorophenyl)-1-(2,2-difluoroethyl)-1H-indazol-5-yl) methyl ketone

[0419]

[0420] Following the synthetic method of Example 20, 6-N-Boc-3,6-diazabicyclo[3.1.1]heptane (43 mg, 0.21 mmol, 1.20 eq) was used as the starting material to obtain a colorless oily intermediate, which was directly used for the next reaction. The intermediate was dissolved in 2 mL of ethyl acetate, and then 1 mL of dioxane hydrochloride solution was added. The mixture was stirred at room temperature for 3 h. After the reaction was completed by TLC, the organic solvent was removed by vacuum distillation, and an appropriate amount of water was added. The pH was adjusted to 7-8 with saturated sodium bicarbonate solution. The mixture was extracted three times with ethyl acetate (15 mL × 3). The organic phases were collected and combined, washed twice with saturated brine, dried over anhydrous sodium sulfate for 0.5 h, and concentrated under reduced pressure to obtain 55 mg of a white powdery solid product, with a yield of 73%.

[0421] 1 H NMR (300MHz, CDCl3) δ8.10(s,1H),7.91(s,1H),7.80(d,J=6.7Hz,1H),7.55(s,2H),7.43(d,J=8.8Hz,2H),6.21(t,J=55.0Hz,1H),4.77(td,J=13.3 ,12.9,4.5Hz,2H),4.15(d,J=13.0Hz,1H),3.89(s,2H),3.68(p,J=14.8,1 4.2Hz,3H),2.75(d,J=8.8Hz,1H),2.26(s,1H),0.85(d,J=7.2Hz,1H)ppm. 13 C NMR (101MHz, CDCl3) δ172.61,144.98,142.01,134.94,134.18,130.45,130.28,128.69,127.49,126.04,125 .66,121.46,120.32,113.79,109.80,56.01,55.59,53.32,51.24(t,J=28.3Hz),49.03,30.33ppm.HRMS(ESI + ):m / z[M+H + Calculated for C 21 H 19 ClF2N4O,417.1294; found 417.1285.

[0422] Test Example: Evaluation of In Vitro Enzyme Activity

[0423] The inhibition rate of the compound against PDE4 enzyme activity and the IC50 were determined using both the IMAP-FP method and the enzyme-coupled method. Details are as follows:

[0424] Method 1 (IMAP-FP method)

[0425] The specific measurement method is as follows:

[0426] (1) Add 40 ml of purified water to 10 ml of 5× IMAP reaction buffer containing 0.1% bovine serum albumin (BSA) provided in the kit, dilute to 1× concentration, then add DTT and bring the final concentration to 1 mM to obtain complete reaction buffer.

[0427] (2) The fluorescein-labeled cAMP with a concentration of 20 μM was diluted at a volume ratio of 1:200, that is, 4.5 μL of fluorescein-labeled cAMP with a concentration of 20 μM was added to 900 μL of reaction buffer to make the final concentration 100 nM.

[0428] (3) PDE4B1 and PDE4D3 were diluted to 15 ng / ml with reaction buffer, and the positive drug rolipram and the compound to be tested were diluted to 10 μM and 1 μM with DMSO.

[0429] (4) Add the following to 120 ml of 5×IMAP binding buffer A and IMAP binding buffer B provided in the kit:

[0430] Dilute 480ml of purified water to a 1× concentration. Mix 17ml of 1× binding buffer A and 3ml of 1× binding buffer B thoroughly, then add 33μL of binding reagent, mix again, and prepare 20ml of stop solution. Store at 4℃ until use.

[0431] (5) Add 10 μL of enzyme dilution and 10 μL of compound solution to a 96-well plate, incubate at room temperature for 15 minutes, and then add...

[0432] 10 μL of fluorescein-labeled cAMP dilution buffer was sealed and protected from light, and incubated at 25°C for 15 minutes. Finally, 60 μL of stop solution was added to each well, including the blank wells. The wells were sealed and protected from light, and incubated at room temperature for 1 hour. The data were then read using an Envision multi-mode microplate reader with the excitation wavelength set to 480 nm and the emission wavelength set to 535 nm. The collected data were converted into inhibition rate values.

[0433] Calculation method:

[0434] Inhibition%=(Max-Signal) / (Max-Min)*100%

[0435] Max: Fluorescence polarization value of the wells containing DMSO and enzyme; Signal: Fluorescence polarization value of the wells containing inhibitor;

[0436] Min: Fluorescence polarization value of wells with and without enzyme, when DMSO is added.

[0437] Method 2 (Enzyme Coupling Method)

[0438] The specific testing method is as follows:

[0439] (1) Preparation of compound solutions:

[0440] ① Preparation of stock solution (10mM): Weigh an appropriate amount of the compound (0.005mmol) and dissolve it in 0.5mL of DMSO. Mix thoroughly by repeatedly blowing or shaking.

[0441] ②1mM solution: Take 100μL of solution ①, add 900μL of DMSO solution, and mix thoroughly by blowing or shaking repeatedly.

[0442] ③ 100μM solution: Take 100μL of solution ②, add 900μL of DMSO solution, and mix thoroughly by blowing or shaking repeatedly.

[0443] ④ 10μM solution: Take 100μL of solution ③, add 900μL of DMSO solution, and mix thoroughly by repeatedly blowing or shaking.

[0444] ⑤ 1μM solution: Take 100μL of solution ④, add 900μL of DMSO solution, and mix thoroughly by blowing and swishing repeatedly.

[0445] ⑥ 100 nM solution: Take 100 μL of solution ⑤, add 900 μL of DMSO solution, and mix thoroughly by blowing and rinsing repeatedly.

[0446] ⑦ 10 nM solution: Take 100 μL of solution ⑥, add 900 μL of DMSO solution, and mix thoroughly by blowing and swishing repeatedly.

[0447] ⑧ 1 nM solution: Take 100 μL of solution ⑦, add 900 μL of DMSO solution, and mix thoroughly by blowing and swishing repeatedly.

[0448] ⑨ 0.1 nM solution: Take 100 μL of solution ⑧, add 900 μL of DMSO solution, and mix thoroughly by blowing and rinsing repeatedly.

[0449] ⑩ 0.01 nM solution: Take 100 μL of solution ⑨, add 900 μL of DMSO solution, and mix thoroughly by blowing and rinsing repeatedly.

[0450] (2) Preparation of mixed solution:

[0451] ① Buffer preparation: Prepare 20 mL of an aqueous solution containing 61 mM Tris-HCl, 12 mM MgCl2, 61 mM KCl, 6.1 mM TCEP, 0.48 mM PEP, 0.012 mM NADH, and 0.048 mM ATP, pH 8.0.

[0452] ② Myokinase stock solution: Dissolve the enzyme in 2 mL of 3.2 M (NH4)2SO4 distilled aqueous solution, pH = 6.0, containing 0.0001 M EDTA, 10% (m / v) glycerol and 0.01% NaN3 as the stock solution. Mix thoroughly by slow pipetting, dispense into 5 tubes, 400 μL / EP tubes. Reconstitute on ice for 5 min before use, then reconstitute at room temperature for 3 min. Mix thoroughly by slow pipetting, and slowly pipet 3 times before sampling.

[0453] ③ Pyruvate kinase stock solution: Dissolve the enzyme in 2 mL of 3.2 M (NH4)2SO4 distilled aqueous solution, pH = 6.0, containing 0.0001 M EDTA, 10% (m / v) glycerol and 0.01% NaN3 as the stock solution. Mix thoroughly by slow pipetting, dispense into 10 tubes or 200 μL / EP tubes, reconstitute on ice for 5 min before use, then reconstitute at room temperature for 3 min, mix thoroughly by slow pipetting, and slowly pipet 3 times before sampling.

[0454] ④ Lactate dehydrogenase stock solution: Dissolve the enzyme in 50 mL of 3.2 M (NH4)2SO4 distilled aqueous solution, pH = 6.0, containing 0.0001 M EDTA, 10% (m / v) glycerol and 0.01% NaN3 as the stock solution. Mix thoroughly by slow pipetting, dispense into 250 tubes or 200 μL / EP tubes. Reconstitute in an ice bath for 5 min before use, then reconstitute at room temperature for 3 min. Mix thoroughly by slow pipetting, and slowly pipet 3 times before sampling.

[0455] ⑤ PDE4B1 stock solution (10 nM): Dissolve 10 μg of enzyme (MW = 109 kDa, purity 70%) in 6.419 mL of buffer (25 mM Tris-HCl, pH 8.0, 100 mM sodium chloride, 0.05% Tween-20, 50% glycerol and 3 mM DTT), gently pipette to mix thoroughly, and divide into 10 tubes, 641.9 μL / tube. Before use, reconstitute on ice for 5 min, then at room temperature for 3 min, gently pipette to mix thoroughly, and gently pipette 3 times before sampling.

[0456] ⑥ PDE4D3 stock solution (10 nM): Dissolve 5 μg of enzyme (MW = 102.4 kDa, purity 70%) in 3.416 mL of buffer (25 mM Tris-HCl, pH 8.0, 100 mM sodium chloride, 0.05% Tween-20, 30% glycerol and 3 mM DTT), gently pipette to mix thoroughly, and divide into 5 tubes, 629.2 μL / tube. Before use, reconstitute on ice for 5 min, then at room temperature for 3 min, gently pipette to mix thoroughly, and gently pipette 3 times before sampling.

[0457] ⑦ Mixed solution 165μL: Take 6μL of myokinase stock solution + 3.2μL of pyruvate kinase stock solution + 2μL of lactate dehydrogenase stock solution + 10μL of PDE4B1 or PDE4D3 stock solution, and make up the remainder with 143.8μL of buffer. Mix thoroughly by slow pipetting.

[0458] (3) Preparation of cAMP solution (0.032 mM):

[0459] Take an appropriate amount of cAMP and prepare 2 mL of a 0.032 mM solution using buffer.

[0460] (4) Operation procedure:

[0461] ① Prepare a mixed solution in a 96-well plate. First, add 143.8 μL of buffer, then add the rest according to the same method. Slowly pipette and mix thoroughly.

[0462] ② Add the compound solution sequentially, gently blow and mix thoroughly, then incubate at 25°C for 5–10 minutes.

[0463] ③ Add cAMP before measurement, blow it three times to mix it evenly, and measure it quickly.

[0464] ④ Use an ELISA reader with an excitation wavelength of 355 nm and an emission wavelength of 460 nm. Continue measuring for 10 minutes, stopping the measurement once a plateau appears. The initial reaction rate should reach -0.7 RFU / s.

[0465] Calculation method:

[0466] Inhibition%=(Max-Signal) / (Max-Min)*100%

[0467] Max: Fluorescence intensity in the well without cAMP; Signal: Fluorescence intensity in the well with inhibitor;

[0468] Min: Fluorescence intensity in the inhibitor-free pore.

[0469] Results of PDE4 inhibitory activity assays for compounds 1–21 and 22–33:

[0470] Table 1. Inhibitory activities of compounds 1–21 against PDE4B and PDE4D (IMAP-FP method)

[0471]

[0472]

[0473] Note: The measured inhibition rates are the average of the two groups.

[0474] Table 2. Inhibitory activities of compounds 22–33 against PDE4B and PDE4D (coupling enzyme method)

[0475]

[0476]

[0477] Evaluation of in vitro anti-inflammatory activity (qRT-PCR method):

[0478] Quantitative real-time (qRT-PCR) detection of mRNA expression

[0479] The steps for qRT-PCR are as follows:

[0480] (1) Spray 75% ethanol on the work surface and wipe it clean. Use tweezers heated with a lighter to pick up the yellow, white, and blue enzyme-free pipette tips and prepare two 1.5ml enzyme-free EP tubes. Remove the cells and place them on ice. Discard the culture medium, add PBS to wash twice, discard the PBS, and then add 600μL of RNAiso Plus to the culture flask. Gently pipette using a 1ml regular pipette tip. Transfer the cell lysis buffer to the enzyme-free EP tube and place it on ice for 15min.

[0481] (2) Add 120 μL of chloroform to the EP tube, vortex to mix, and place on ice for 5 min. Centrifuge at 12000g for 15 min at 4°C. At this point, the liquid in the tube, from top to bottom, consists of: a colorless supernatant containing RNA, a white protein layer containing DNA, and an organic liquid layer. Transfer 100 μL of the supernatant to another EP tube, add 100 μL of isopropanol, invert the EP tube to mix thoroughly, and let stand at room temperature for 10 min. Centrifuge at 12000g for 10 min at 4°C, and discard the supernatant to obtain the RNA precipitate. Then add 600 μL of pre-cooled 75% ethanol, gently shake to wash the precipitate, centrifuge at 7500g for 5 min at 4°C, and discard the supernatant. When the EP tube has dried completely at room temperature and the precipitate is translucent, add 20 μL of DEPC water to dissolve the precipitate.

[0482] (3) After obtaining the total RNA solution, the RNA purity was analyzed by absorbance method. The UV spectrophotometer was turned on, and the probe was cleaned twice with anhydrous ethanol and twice with DEPC water. DEPC water was added to zero the instrument, and 2 μL of each sample was taken to detect the RNA concentration. Based on the detected RNA concentration, the concentration of each group of samples was adjusted to approximately 400 ng / μL with DEPC water.

[0483] (4) The primers were designed and obtained through commercial purchase.

[0484] Table 3. Sequences of target gene and internal reference primers

[0485]

[0486] Following the instructions for the EvoM-MLV RT Kit with gDNA Clean for qPCR, prepare the genomic removal reaction mixture in a standard volume of 10 μL. All procedures are performed on ice to remove residual genomic DNA.

[0487] The system was reacted at 42℃ for 2 min.

[0488] Next, the extracted total RNA was reverse transcribed into cDNA. All samples were placed in an RNA reverse transcriptase instrument and incubated at 37°C for 15 min, then at 85°C for 5 s. After cooling, they were temporarily stored at 4°C.

[0489] Quantitative real-time polymerase chain reaction

[0490] (6) Real-time quantitative polymerase chain reaction (qRT-PCR) was used with GAPDH as a standardized internal control to detect the mRNA expression levels of TNF-α in bv2 cells of the blank group, model group, and drug-treated group. cDNAs reverse-transcribed from each concentration were diluted to the same concentration and then amplified by real-time quantitative PCR. Amplification conditions: 95℃ pre-denaturation for 10 min, one cycle, followed by 95℃ for 15 min, annealing at 60℃ for 20 min, and extension at 72℃ for 20 min, for 40 cycles. Data analysis of qRT-PCR results was performed using the 2-ΔΔCt method.

[0491] The method involves performing three technical replicates for each gene.

[0492] Experimental results:

[0493] Experimental data were processed and analyzed using Graphpad Prism 9.0. All data are expressed as mean ± standard deviation, and significance was indicated by P < 0.05.

[0494] The results of compounds 5 and 12 inhibiting LPS-induced expression of the inflammatory cytokine (TNF-α) in BV2 cells are as follows: Figure 1 As shown.

[0495] qRT-PCR analysis of TNF-α expression levels in BV2 cell culture supernatant showed that compound 5 significantly inhibited LPS-induced TNF-α mRNA expression. Compound 5 began to exhibit neuroprotective effects at a concentration of 15 μM, with a reduced TNF-α mRNA expression level compared to the LPS-induced model group. Compound 12 reduced TNF-α mRNA expression in a dose-dependent manner, indicating that it effectively inhibited LPS-induced inflammatory responses in BV2 cells.

Claims

1. A compound as shown in Formula I-3 or a pharmaceutically acceptable salt thereof, in, R 2 For not replaced or by R 2-4 Substituted phenyl, unsubstituted or R 2-5 The substituted 5-6 heteroaryl group, wherein the 5-6 heteroaryl group is thienyl or pyridinyl; R 3 For H; R 4 It is an unsubstituted 3-6 membered cycloalkyl group; Or, R 3 and R 4 Together with the nitrogen atom attached to it, it forms an unsubstituted or R-shaped structure. 3-4 The substituted 3-12-membered heterocyclic alkyl group, wherein the 3-12-membered heterocyclic alkyl group is azahexacyclobutane, morpholino, pyrrolidone, piperidinyl, piperazine, etc. , , , , , , or ; R 2-4 and R 2-5 Each is independently a halogen, hydroxyl, amino, cyano, C 1-4 Alkyl, C 1-4 Alkoxy, C 1-4 Halogenated alkyl or C 1-4 Halogenated alkoxy groups; R 3-4 Halogen, hydroxyl, C 1-4 Haloalkyl, C 1-4 Alkyl, C 1-4 Alkoxy or C 1-4 Halogenated alkoxy groups.

2. The compound of formulas I-3 according to claim 1, or a pharmaceutically acceptable salt thereof, wherein, The compound shown in Formula I-3 or its pharmaceutically acceptable salt is the compound shown in Formula I-1A or its pharmaceutically acceptable salt. Wherein, ring A is unsubstituted or replaced by R. 3-4 The substituted 3-12-membered heterocyclic alkyl group, wherein the 3-12-membered heterocyclic alkyl group is azahexacyclobutane, morpholino, pyrrolidone, piperidinyl, piperazine, etc. , , , , , , or R 2 and R 3-4 As defined in claim 1.

3. The compound of formulas I-3 according to claim 1, or a pharmaceutically acceptable salt thereof, wherein, The compound shown in Formula I-3 or its pharmaceutically acceptable salt is the compound shown in Formula I-2A or its pharmaceutically acceptable salt. Among them, R 3’ For H; R 4’ R is an unsubstituted 3-6 membered cycloalkyl group. 2 As defined in claim 1.

4. Any of the following structural compounds or their pharmaceutically acceptable salts, 。 5. A method for preparing the compound of formulas I-3 according to claim 1, comprising: The compound shown in Formula II is condensed with the compound shown in Formula III to obtain the compound shown in Formula I. The compound shown in Formula I is the same as the compound shown in Formula I-3. Among them, R 2 R 3 and R 4 As defined in claim 1.

6. A pharmaceutical composition comprising the compound or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient according to any one of claims 1-4.

7. Use of a compound according to any one of claims 1-4, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 6, in the preparation of a medicament as a PDE4 inhibitor.

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