Bicyclic compound and use thereof

Abstract

Provided are a compound represented by formula (I), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, a pharmaceutical composition comprising same, and a use thereof as an allosteric modulator of β-glucocerebrosidase (GCase), preferably for treating a neurodegenerative disease.

Description

Bicyclic compounds and their applications

[0001] Cross-reference of related applications

[0002] This application claims priority and benefit to Chinese patent applications No. 202510290385.8 filed with the China National Intellectual Property Administration (CNIPA) on March 12, 2025; Chinese patent applications No. 202510525057.1 filed with the CNIPA on April 24, 2025; Chinese patent applications No. 202510717035.5 filed with the CNIPA on May 30, 2025; and Chinese patent applications No. 202511069162.5 filed with the CNIPA on July 31, 2025, the contents of which are incorporated herein by reference in their entirety. Technical Field

[0003] This disclosure pertains to the field of pharmaceutical technology, specifically relating to bicyclic compounds as allosteric regulators of β-glucocerebrosidase (also known as acid β-glucosidase, D-glucosyl-N-acylsphingosine glucosylhydrolase, or GCase), pharmaceutical compositions containing such bicyclic compounds, and their use in the prevention or treatment of neurodegenerative diseases. Background Technology

[0004] β-Glucocephalinase is an enzyme with glucosylceramidinase activity that metabolizes glycosphingolipid glucosylceramide (GlcCer) into ceramide and glucose. β-Glucocephalinase exhibits maximum activity at pH 5.5 (the pH of the lysosomal compartment). Within the lysosome, it remains membrane-bound and degrades its substrate, glucocerebroside (GluCer). It requires the activation of the protein Saposin C and negatively charged lipids to achieve maximum catalytic activity, ensuring the normal functioning of cellular protein degradation and cell signaling.

[0005] Mutations in GBA1 (the gene encoding the lysosomal enzyme glucocerebrosidase) are one of the most common known genetic risk factors for the development of Parkinson's disease and related synucleinopathies, and also a cause of the rare autosomal storage disorder Gaucher disease. GBA1 mutations can lead to protein degradation, disruption of lysosomal targeting, and reduced enzyme performance in lysosomes. Gaucher disease is phenotypic, presenting in both neuropathic and non-neuropathic forms. Patients with Gaucher disease and heterozygous carriers have an increased risk of developing Parkinson's disease and Lewy body dementia. Reduced GCase activity levels have been detected in different brain regions of Parkinson's disease patients. Furthermore, studies have shown that GCase modulators can restore lysosomal function, reduce lipid substrate accumulation, and thus inhibit pathological α-Syn accumulation. Therefore, GCase modulators are considered a strategy for developing new drugs to alleviate and treat Parkinson's disease.

[0006] Therefore, researching novel, potent, and highly selective GCase modulators is of great significance for the treatment of neurodegenerative diseases. Summary of the Invention

[0007] This invention provides a compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof:

[0008] in,

[0009] The ring C is selected from the following structural groups:

[0010] Indicates a single bond or a double bond;

[0011] U is selected from C=O, N, and CR. 2 ;

[0012] Q is selected from C and N;

[0013] V is selected from O, S, NR 3 and CHR 2 ;

[0014] t' is 1 or 2;

[0015] s' is selected from 0, 1, or 2;

[0016] R 5 Selected from halogens, =O, C1-C6 alkyl, C1-C6 haloalkyl or C1-C4 alkoxy;

[0017] R c Selected from C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl or 4-6 membered heterocyclic groups, wherein each of the C1-C6 alkyl, C3-C6 cycloalkyl or 4-6 membered heterocyclic groups is independently and optionally substituted by NH2, OH, halogen or C1-C6 alkyl.

[0018] R 2 Selected from H, halogens, C1-C6 alkyl groups, C1-C6 haloalkyl groups, or C1-C4 alkoxy groups;

[0019] R 3 Selected from H, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C4 alkoxy;

[0020] L and W are each independently selected from the key and -CR. a R b -、C(O), O、S、-CR a R b O- or NR a ;

[0021] R a R b It is independently selected from H, halogen, C1-C6 alkyl, C1-C6 haloalkyl or C1-C4 alkoxy;

[0022] Ring B is selected from the following groups:

[0023] a)C6-C 14 Spirocycloalkylene, C6-C 14 fused cycloalkylene, C5-C 12 Bridged cycloalkyl groups, 6-14 membered spirocycloalkyl groups, 6-14 membered fused heterocyclic groups, or 5-12 membered bridged heterocyclic groups;

[0024] or,

[0025] b)

[0026] Each R 4 It is independently selected from deuterium, OH, halogen, =O, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy or C3-C6 cycloalkyl;

[0027] Ring A is selected from C3-C 12 Cycloalkyl, 4-12 membered heterocyclic, 5-10 membered heteroaryl or C6-C 10 Aryl;

[0028] Each R 1 The group is independently selected from CN, NH2, OH, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C3-C6 cycloalkyl or 4-10 heterocyclic group, wherein each of the C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl or 4-10 heterocyclic group is independently and optionally substituted by NH2, OH, halogen or C1-C6 alkyl;

[0029] n is selected from 0, 1, or 2;

[0030] p and q are independently selected from 0, 1, or 2;

[0031] The condition is that when ring C is selected from When L is selected from -CR a R b -, C(O), O or S.

[0032] In some implementations, in formula (I),

[0033] The ring C is selected from the following structural groups:

[0034] R cSelected from C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl or 4-6 membered heterocyclic groups, wherein each of the C1-C6 alkyl, C3-C6 cycloalkyl or 4-6 membered heterocyclic groups is independently and optionally substituted by NH2, OH, halogen or C1-C6 alkyl.

[0035] R 2 Selected from H, halogens, C1-C6 alkyl groups, C1-C6 haloalkyl groups, or C1-C4 alkoxy groups;

[0036] R 3 Selected from H, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C4 alkoxy;

[0037] L and W are each independently selected from the key and -CR. a R b - C(O), O, S or CH2O;

[0038] R a R b It is independently selected from H, halogen, C1-C6 alkyl, C1-C6 haloalkyl or C1-C4 alkoxy;

[0039] Ring B is selected from the following groups:

[0040] a)C6-C 14 Spirocycloalkylene, C6-C 14 fused cycloalkylene, C5-C 12 Bridged cycloalkyl groups, 6-14 membered spirocycloalkyl groups, 6-14 membered fused heterocyclic groups, or 5-12 membered bridged heterocyclic groups;

[0041] or,

[0042] b)

[0043] Each R 4 It is independently selected from deuterium, OH, halogen, =O, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy or C3-C6 cycloalkyl;

[0044] Ring A is selected from C3-C 12 Cycloalkyl, 4-12 membered heterocyclic, 5-10 membered heteroaryl or C6-C 10 Aryl;

[0045] Each R 1The group is independently selected from CN, NH2, OH, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C3-C6 cycloalkyl or 4-10 heterocyclic group, wherein each of the C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl or 4-10 heterocyclic group is independently and optionally substituted by NH2, OH, halogen or C1-C6 alkyl;

[0046] n is selected from 0, 1, or 2;

[0047] p and q are independently selected from 0, 1, or 2;

[0048] The condition is that when ring C is selected from When L is selected from -CR a R b -, C(O), O or S.

[0049] In some implementations, in formula (I),

[0050] The ring C is selected from the following structural groups:

[0051] R c Selected from C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl or 4-6 membered heterocyclic groups, wherein each of the C1-C6 alkyl, C3-C6 cycloalkyl or 4-6 membered heterocyclic groups is independently and optionally substituted by NH2, OH, halogen or C1-C6 alkyl.

[0052] R 2 Selected from H, halogens, C1-C6 alkyl groups, C1-C6 haloalkyl groups, or C1-C4 alkoxy groups;

[0053] R 3 Selected from H, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C4 alkoxy;

[0054] L and W are each independently selected from the key and -CR. a R b -, C(O), O or S;

[0055] R a R b It is independently selected from H, halogen, C1-C6 alkyl, C1-C6 haloalkyl or C1-C4 alkoxy;

[0056] Ring B is selected from the following groups:

[0057] a)C6-C 14 Spirocycloalkylene, C6-C 14 fused cycloalkylene, C5-C 12Bridged cycloalkyl groups, 6-14 membered spirocycloalkyl groups, 6-14 membered fused heterocyclic groups, or 5-12 membered bridged heterocyclic groups;

[0058] or,

[0059] b)

[0060] Each R 4 It is independently selected from deuterium, OH, halogen, =O, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy or C3-C6 cycloalkyl;

[0061] Ring A is selected from C3-C 12 Cycloalkyl, 4-12 membered heterocyclic, 5-10 membered heteroaryl or C6-C 10 Aryl;

[0062] Each R 1 The group is independently selected from CN, NH2, OH, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C3-C6 cycloalkyl or 4-10 heterocyclic group, wherein each of the C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl or 4-10 heterocyclic group is independently and optionally substituted by NH2, OH, halogen or C1-C6 alkyl;

[0063] n is selected from 0, 1, or 2;

[0064] p and q are independently selected from 0, 1, or 2;

[0065] The condition is that when ring C is selected from When L is selected from -CR a R b -, C(O), O or S.

[0066] In some embodiments, the compound of formula (I) or its stereoisomer or a pharmaceutically acceptable salt thereof is selected from the compound of formula (I-1) or its stereoisomer or a pharmaceutically acceptable salt thereof.

[0067] Wherein, U is selected from N and CR 2 ;

[0068] Ring A, Ring B, W, L, V, R 1 R 4 R c n, p, q are defined as above.

[0069] In some implementations, U is N.

[0070] In some implementations, U is CR 2 .

[0071] In some implementations, V is NR 3 .

[0072] In some implementations, U is N and V is NR. 3 .

[0073] In some implementations, U is CR 2 And V is NR 3 .

[0074] In some embodiments, the compound of formula (I) or its stereoisomer or a pharmaceutically acceptable salt thereof is selected from the compound of formula (I-1B) or its stereoisomer or a pharmaceutically acceptable salt thereof.

[0075] Among them, rings A, B, W, L, and R 1 R 2 R 3 R 4 R c p, q, n are defined as above.

[0076] In some embodiments, the compound of formula (I) or its stereoisomer or a pharmaceutically acceptable salt thereof is selected from the compound of formula (I-1B-1) or its stereoisomer or a pharmaceutically acceptable salt thereof.

[0077] Among them, rings A, B, W, L, and R 1 R 2 R 3 R 4 R c p and q are defined as above.

[0078] In some embodiments, the compound of formula (I) or its stereoisomer or a pharmaceutically acceptable salt thereof is selected from the compound of formula (I-1A) or its stereoisomer or a pharmaceutically acceptable salt thereof.

[0079] Among them, rings A, B, W, L, and R 1 R 3 R 4 R c p and q are defined as above.

[0080] In some embodiments, the compound of formula (I) or its stereoisomer or a pharmaceutically acceptable salt thereof is selected from the compound of formula (I-1A-1) or its stereoisomer or a pharmaceutically acceptable salt thereof.

[0081] X1 is selected from O, S or CR. 6 R 7 X2 is selected from CR 8 Or N; X3 is selected from CR 9 Or N;

[0082] R 6 R 7 R 8 R 9 It is independently selected from H, deuterium, NH2, OH, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy or C3-C6 cycloalkyl;

[0083] s, m, and r are independently selected from 0, 1, or 2;

[0084] Each R 4 It is independently selected from deuterium, OH, halogen, =O, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy or C3-C6 cycloalkyl;

[0085] q' is selected from 0 or 1;

[0086] "q" is selected from 0 or 1;

[0087] Rings A and R 1 p, R 3 R c As defined above.

[0088] In some embodiments, the compound of formula (I) or its stereoisomer or a pharmaceutically acceptable salt thereof is selected from the compound of formula (I-1A-2) or its stereoisomer or a pharmaceutically acceptable salt thereof.

[0089] Among them, rings A and R 1 p, R 3 R c As defined above.

[0090] In some embodiments, the compound of formula (I) or its stereoisomer or a pharmaceutically acceptable salt thereof is selected from the compound of formula (I-2) or its stereoisomer or a pharmaceutically acceptable salt thereof.

[0091] Among them, rings A, B, W, L, V, and R 1 R 4 R 5 R c The definitions of n, p, q, t', and s' are as described above.

[0092] In some implementations, V is selected from O and NR. 3 and CHR2 .

[0093] In some implementations, t' is 1.

[0094] In some implementations, t' is 2.

[0095] In some embodiments, the compound of formula (I) or its stereoisomer or a pharmaceutically acceptable salt thereof is selected from the compound of formula (I-2A) or its stereoisomer or a pharmaceutically acceptable salt thereof.

[0096] Among them, rings A, B, W, L, V, and R 1 R 4 R c n, p, q are defined as above.

[0097] In some implementations, V is selected from NR. 3 or CHR 2 .

[0098] In some implementations, V is NR 3 .

[0099] In some embodiments, the compound of formula (I) or its stereoisomer or a pharmaceutically acceptable salt thereof is selected from the compound of formula (I-2B) or its stereoisomer or a pharmaceutically acceptable salt thereof.

[0100] Among them, rings A, B, W, L, V, and R 1 R 4 R c n, p, q are defined as above.

[0101] In some implementations, V is selected from O or CHR. 2 .

[0102] In some implementations, V is selected from O or CH2. In some implementations, V is CH2.

[0103] In some embodiments, the cyclic C is selected from the following structural groups:

[0104] In some embodiments, the cyclic C is selected from the following structural groups:

[0105] In some embodiments, the cyclic C is selected from the following structural groups:

[0106] In some embodiments, the cyclic C is selected from the following structural groups:

[0107] In some embodiments, the cyclic C is selected from the following structural groups:

[0108] In some embodiments, the cyclic C is selected from the following structural groups:

[0109] In some embodiments, the cyclic C is selected from the following structural groups:

[0110] In some embodiments, the cyclic C is selected from the following structural groups:

[0111] In some embodiments, the cyclic C is selected from the following structural groups:

[0112] In some implementation schemes, It is a single bond, U is C=O, and Q is N.

[0113] In some implementation schemes, It is a double bond, and U is selected from N or CR. 2 Q is C.

[0114] In some implementations, U is selected from C=O, N, and CH.

[0115] In some implementation schemes, R 5 =O.

[0116] In some implementations, s' is 0 or 1.

[0117] In some implementation schemes, R c Selected from C1-C6 alkyl or 4-6 membered heterocyclic groups, wherein each of the C1-C6 alkyl or 4-6 membered heterocyclic groups is optionally substituted independently with NH2, OH, halogen or C1-C6 alkyl.

[0118] In some implementation schemes, R c Selected from C1-C6 alkyl groups, wherein the C1-C6 alkyl groups are optionally substituted with NH2, OH, halogens or C1-C6 alkyl groups.

[0119] In some implementation schemes, R c Selected from C1-C6 alkyl groups, wherein the C1-C6 alkyl groups are optionally substituted with halogens.

[0120] In some implementation schemes, R c Selected from C1-C6 alkyl groups, wherein the C1-C6 alkyl groups are optionally substituted with F.

[0121] In some implementation schemes, R c Selected from CH3, CH2CH3, CH2CHF2, CH2CF2CH3, CH2CF3 or oxocyclic butyl.

[0122] In some implementation schemes, R c Selected from CH3, CH2CH3, CH2CHF2, CH2CF2CH3, CH2CF3 or

[0123] In some implementation schemes, R c Selected from CH3, CH2CH3, CH2CHF2, CH2CF2CH3 or oxocyclic butyl.

[0124] In some implementation schemes, R c Selected from CH3 or CH2CHF2.

[0125] In some implementation schemes, R c Selected from CH2CHF2.

[0126] In some implementation schemes, R 2 Selected from H or C1-C6 alkyl groups.

[0127] In some implementation schemes, R 2 Selected from H or CH3.

[0128] In some implementation schemes, R 2 For H.

[0129] In some implementation schemes, R 3 Selected from H or C1-C6 alkyl groups.

[0130] In some implementation schemes, R 3 Selected from H or CH3.

[0131] In some implementation schemes, R 3 It is CH3.

[0132] In some embodiments, the cyclic C is selected from the following structural groups:

[0133] In some implementation schemes, L and W are each independently selected from the key and -CR. a R b -, C(O), O, S, CH2O or NR a .

[0134] In some implementation schemes, L and W are each independently selected from the key and -CR. a R b - C(O), O, S or CH2O.

[0135] In some implementation schemes, L and W are each independently selected from the key and -CR. a R b -, C(O), O or S.

[0136] In some implementations, L is selected from bond, -CH2-, C(O), O, S, NH or CH2O.

[0137] In some implementations, L is selected from a bond, -CH2-, C(O), O, or S.

[0138] In some implementations, L is selected from bond, -CH2-, O, CH2O or NH.

[0139] In some implementations, L is selected from a bond, -CH2-, or O.

[0140] In some implementations, L stands for key.

[0141] In some implementations, L stands for -CH2-.

[0142] In some implementations, L is O.

[0143] In some implementations, W is selected from bond, -CH2-, C(O), O, S, or CH2O.

[0144] In some implementations, W is selected from bond, -CH2-, C(O), O, or S.

[0145] In some implementations, W is selected from bonds or CH2O.

[0146] In some implementations, W is the key.

[0147] In some implementations, W is CH2O.

[0148] In some implementations, L is selected from bond, -CH2- or O, and W is selected from bond or CH2O.

[0149] In some implementations, L is a key, and W is selected from key, -CR a R b -, C(O), O or S.

[0150] In some implementations, L is a bond, and W is selected from either a bond or CH2O.

[0151] In some implementations, L is selected from key, -CR aR b -, O, CH2O or NR a And W is the key.

[0152] In some implementations, L is selected from bond, -CH2-, O, CH2O or NH, and W is bond.

[0153] In some implementations, L is selected from key, -CR a R b - C(O), O or S, and W is a bond.

[0154] In some implementations, L is -CH2-, and W is selected from bonds, -CR a R b -, C(O), O or S.

[0155] In some implementations, L is a bond, -CH2-, or O, and W is a bond.

[0156] In some implementations, L is a bond and W is CH2O.

[0157] In some implementations, L is O and W is a bond.

[0158] In some implementations, L is -CH2- and W is a bond.

[0159] In some implementations, L is the key and W is the key.

[0160] In some implementation schemes, R a R b It is independently selected from H or C1-C6 alkyl groups.

[0161] In some implementation schemes, R a R b All are H.

[0162] In some embodiments, ring B is selected from 6-14 spiro-heterocyclic groups, 6-14 fused-heterocyclic groups, or 5-12 bridged-heterocyclic groups.

[0163] In some implementations, ring B is selected from 6-14 nucleotide bicyclic spirocyclic subheterocyclic groups.

[0164] In some implementations, ring B is selected from 3-membered / 6-membered bicyclic spirocyclic subheterocyclic groups, 4-membered / 4-membered bicyclic spirocyclic subheterocyclic groups, 4-membered / 5-membered bicyclic spirocyclic subheterocyclic groups, 4-membered / 6-membered bicyclic spirocyclic subheterocyclic groups, 5-membered / 5-membered bicyclic spirocyclic subheterocyclic groups, or 5-membered / 6-membered bicyclic spirocyclic subheterocyclic groups.

[0165] In some implementations, ring B is selected from 5-membered / 5-membered bicyclic spirocyclic subheterocyclic groups, 4-membered / 4-membered bicyclic spirocyclic subheterocyclic groups, 4-membered / 5-membered bicyclic spirocyclic subheterocyclic groups, 5-membered / 6-membered bicyclic spirocyclic subheterocyclic groups, or 4-membered / 6-membered bicyclic spirocyclic subheterocyclic groups.

[0166] In some implementations, ring B is selected from 4-membered / 4-membered bicyclic spirocyclic subheterocyclic groups, 4-membered / 5-membered bicyclic spirocyclic subheterocyclic groups, 5-membered / 6-membered bicyclic spirocyclic subheterocyclic groups, or 4-membered / 6-membered bicyclic spirocyclic subheterocyclic groups.

[0167] In some implementations, ring B is selected from 4-membered / 5-membered bicyclic spirocyclic subheterocyclic groups, 5-membered / 6-membered bicyclic spirocyclic subheterocyclic groups, or 4-membered / 6-membered bicyclic spirocyclic subheterocyclic groups.

[0168] In some implementations, ring B is selected from 6-14 membered bicyclic fused heterocyclic groups.

[0169] In some implementations, ring B is selected from 3-membered / 5-membered bicyclic fused heterocyclic groups, 6-membered / 6-membered bicyclic fused heterocyclic groups, 5-membered / 6-membered bicyclic fused heterocyclic groups, or 5-membered / 5-membered bicyclic fused heterocyclic groups.

[0170] In some implementations, ring B is selected from 6-membered / 6-membered bicyclic fused heterocyclic groups, 5-membered / 6-membered bicyclic fused heterocyclic groups, or 5-membered / 5-membered bicyclic fused heterocyclic groups.

[0171] In some implementations, ring B is selected from 3-membered / 5-membered bicyclic fused heterocyclic groups, 5-membered / 6-membered bicyclic fused heterocyclic groups, or 5-membered / 5-membered bicyclic fused heterocyclic groups.

[0172] In some implementations, ring B is selected from 5-membered / 5-membered bicyclic fused heterocyclic groups.

[0173] In some implementations, ring B is a 5-12 quinary bicyclic bridged subheterocyclic group.

[0174] In some implementations, ring B is a 6-9 quintic bicyclic bridged subheterocyclic group.

[0175] In some implementations, ring B is a 7- or 8-membered bicyclic bridged subheterocyclic group.

[0176] In some embodiments, the 6-14 membered spiro-heterocyclic group, the 6-14 membered fused-heterocyclic group, or the 5-12 membered bridged-heterocyclic group each independently contains one, two, or three heteroatoms independently selected from N, O, or S.

[0177] In some embodiments, the 6-14 membered spiro-heterocyclic group, the 6-14 membered fused-heterocyclic group, or the 5-12 membered bridged-heterocyclic group each independently contains one, two, or three heteroatoms independently selected from N or O.

[0178] In some embodiments, the 6-14 membered spiro-heterocyclic group, the 6-14 membered fused-heterocyclic group, or the 5-12 membered bridged-heterocyclic group each independently contains one or two heteroatoms independently selected from N.

[0179] In some embodiments, the 3-membered / 6-membered bicyclic spiro-heterocyclic group, the 4-membered / 4-membered bicyclic spiro-heterocyclic group, the 4-membered / 5-membered bicyclic spiro-heterocyclic group, the 4-membered / 6-membered bicyclic spiro-heterocyclic group, the 5-membered / 5-membered bicyclic spiro-heterocyclic group, the 5-membered / 6-membered bicyclic spiro-heterocyclic group, the 3-membered / 5-membered bicyclic fused heterocyclic group, the 6-membered / 6-membered bicyclic fused heterocyclic group, the 5-membered / 6-membered bicyclic fused heterocyclic group, the 5-membered / 5-membered bicyclic fused heterocyclic group, the 6-9-membered bicyclic bridged heterocyclic group, or the 7-membered or 8-membered bicyclic bridged heterocyclic group each independently contains one, two, or three heteroatoms independently selected from N or O.

[0180] In some embodiments, ring B is selected from the following structural groups:

[0181] X1 is selected from O, S or CR. 6 R 7 X2 is selected from CR 8 Or N; X3 is selected from CR 9 Or N;

[0182] R 6 R 7 R 8 R 9 It is independently selected from H, deuterium, NH2, OH, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy or C3-C6 cycloalkyl;

[0183] s, m, and r are independently selected from 0, 1, or 2;

[0184] Each R 4 It is independently selected from deuterium, OH, halogen, =O, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy or C3-C6 cycloalkyl;

[0185] q' is selected from 0 or 1;

[0186] "q" is selected from 0 or 1.

[0187] In some implementations, X1 is selected from O or CR. 6 R 7 .

[0188] In some implementations, X1 is CR 6 R 7 In some implementations, X1 is CH2.

[0189] In some implementations, at least one of X2 and X3 is N.

[0190] In some implementations, X2 is selected from CH or N.

[0191] In some implementations, X3 is selected from CH or N.

[0192] In some implementation schemes, R 6 R 7 R 8 R 9 It is independently selected from H, deuterium or C1-C6 alkyl.

[0193] In some implementation schemes, R 6 R 7 R 8 R 9 Each of them is H independently.

[0194] In some implementations, s, m, and n are independently selected from 0 or 1.

[0195] In some implementations, q' is 0; q” is 0.

[0196] In some embodiments, ring B is selected from the following structural groups:

[0197] Z1 is selected from CR 10 Or N; Z2 is selected from CR 10 Or N;

[0198] R 10 Selected from H, deuterium, NH2, OH, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy or C3-C6 cycloalkyl;

[0199] t, f, g, and k are independently selected from 0, 1, or 2;

[0200] Each R 4 It is independently selected from deuterium, OH, halogen, =O, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy or C3-C6 cycloalkyl;

[0201] q' is selected from 0 or 1;

[0202] "q" is selected from 0 or 1.

[0203] In some implementation schemes, R 10 Selected from H, deuterium, or C1-C6 alkyl groups.

[0204] In some implementation schemes, R 10 For H.

[0205] In some implementations, at least one of Z1 and Z2 is N.

[0206] In some implementations, Z1 is N, and Z2 is selected from CH or N.

[0207] In some implementations, Z1 is CH or N, and Z2 is selected from N.

[0208] In some implementations, Z1 is N and Z2 is CH. In some implementations, Z1 is CH and Z2 is selected from N. In some implementations, Z1 is N and Z2 is N.

[0209] In some implementations, t is 1.

[0210] In some implementations, k is 1.

[0211] In some implementations, f and g are selected from 0 or 1.

[0212] In some implementations, both f and g are 0.

[0213] In some implementations, both f and g are 1.

[0214] In some implementations, q' is 0 and q” is 0.

[0215] In some embodiments, ring B is selected from the following structural groups:

[0216] Among them, Y1 and Y2 are independently selected from N or CH;

[0217] Z is selected from CH2 or CH2CH2;

[0218] Each R 4 It is independently selected from deuterium, OH, halogen, =O, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy or C3-C6 cycloalkyl;

[0219] q is selected from 0, 1 or 2.

[0220] In some implementations, at least one of Y1 and Y2 is N.

[0221] In some implementations, Y1 and Y2 are both CH independently. In some implementations, Y1 and Y2 are both N independently.

[0222] In some implementation schemes, ring B is selected from...

[0223] In some implementation schemes, ring B is selected from...

[0224] In some embodiments, ring B is selected from the following structural groups:

[0225] In some implementation schemes, ring B is selected from...

[0226] In some implementation schemes, ring B is selected from...

[0227] In some implementation schemes, ring B is selected from...

[0228] In some implementations, ring A is selected from C3-C 10 Cycloalkyl, 4-9 membered heterocyclic, 5-6 membered heteroaryl or phenyl.

[0229] In some implementations, ring A is a 5-10 membered heteroaryl or phenyl group.

[0230] In some implementations, ring A is a 5-6 membered heteroaryl or phenyl group.

[0231] In some implementations, ring A is a 5-10 membered heteroaryl group.

[0232] In some implementations, ring A is a 5-6 membered heteroaryl group.

[0233] In some embodiments, ring A is phenyl, pyridyl, pyrimidinyl, or pyridazinyl.

[0234] In some embodiments, ring A is pyridinyl, pyrimidinyl, or pyridazinyl.

[0235] In some embodiments, ring A is pyridinyl, pyrimidinyl, or pyrazolyl.

[0236] In some implementations, ring A is pyridinyl.

[0237] In some implementations, ring A is a phenyl group.

[0238] In some implementations, ring A is selected from...

[0239] In some implementations, ring A is selected from...

[0240] In some implementations, ring A is selected from...

[0241] In some implementations, ring A is selected from...

[0242] In some implementations, ring A is

[0243] In some implementation schemes, R 1 Selected from CN, halogen, C1-C6 alkyl, C1-C6 haloalkyl or C1-C6 alkoxy, wherein each of the C1-C6 alkyl or C1-C6 alkoxy is optionally substituted independently by NH2, OH, halogen or C1-C6 alkyl.

[0244] In some implementation schemes, R 1 Selected from halogens, C1-C6 alkyl groups, C1-C6 haloalkyl groups, or C1-C6 alkoxy groups, wherein each of the C1-C6 alkyl groups or C1-C6 alkoxy groups is optionally substituted independently by NH2, OH, halogens, or C1-C6 alkyl groups.

[0245] In some implementation schemes, R 1 The components are selected from halogens, C1-C3 alkyl groups, C1-C3 haloalkyl groups, or C1-C3 alkoxy groups, wherein each of the C1-C3 alkyl groups or C1-C3 alkoxy groups is optionally substituted independently by NH2, OH, halogens, or C1-C6 alkyl groups.

[0246] In some implementation schemes, R 1 It is selected from halogens, C1-C6 alkyl groups, or C1-C6 haloalkyl groups.

[0247] In some implementation schemes, R 1 Selected from CN, F, Cl, CH3, CHF2, CF3 or -OCHF2.

[0248] In some implementation schemes, R 1 Selected from F, Cl, CH3, CHF2, CF3 or -OCHF2.

[0249] In some implementation schemes, R 1 Selected from C1-C6 haloalkyl groups.

[0250] In some implementation schemes, R 1 It is CF3.

[0251] In some implementation schemes, Selected from the following structures:

[0252] In some implementation schemes, Selected from the following structures:

[0253] In some implementation schemes, Selected from the following structures:

[0254] In some implementation schemes, Selected from the following structures:

[0255] In some implementation schemes, It has the following structure:

[0256] In some implementation schemes, R 4 It is selected from deuterium, OH, halogen, =O, C1-C6 alkyl or C1-C6 haloalkyl.

[0257] In some implementation schemes, R 4 Selected from deuterium, =O or C1-C6 alkyl groups.

[0258] In some implementation schemes, R 4 Selected from deuterium, =O, or CH3.

[0259] In some implementation schemes, R 4 Selected from deuterium or =O.

[0260] In some implementation schemes, R 4 =O.

[0261] In some implementations, p is selected from 0 or 1. In some implementations, p is 1.

[0262] In some implementations, q is selected from 0 or 1. In some implementations, q is 0.

[0263] In some implementations, n is 1 or 2.

[0264] In some implementations, n is 1.

[0265] In some embodiments, the compound of formula (I) or its isomer or pharmaceutically acceptable salt thereof is selected from the following compounds or their isomers or pharmaceutically acceptable salts:

[0266] On the other hand, this disclosure provides pharmaceutical compositions comprising a compound of formula (I) or formula (I-1) or formula (I-1B) or formula (I-1B-1) or formula (I-1A) or formula (I-1A-1) or formula (I-1A-2) or formula (I-2) or formula (I-2A) or formula (I-2B) of this disclosure, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0267] On the other hand, this disclosure provides a method for preventing or treating GCase-mediated diseases in mammals, comprising administering to a mammal, preferably a human, a therapeutically effective amount of a compound of formula (I) or formula (I-1) or formula (I-1B) or formula (I-1B-1) or formula (I-1A) or formula (I-1A-1) or formula (I-1A-2) or formula (I-2) or formula (I-2A) or formula (I-2B) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0268] On the other hand, this disclosure provides the use of a compound of formula (I) or formula (I-1) or formula (I-1B) or formula (I-1B-1) or formula (I-1A) or formula (I-1A-1) or formula (I-1A-2) or formula (I-2) or formula (I-2A) or formula (I-2B) or its stereoisomer or pharmaceutically acceptable salt or pharmaceutical composition thereof in the preparation of a medicament for the prevention or treatment of GCase-mediated diseases.

[0269] On the other hand, this disclosure provides the use of compounds of formula (I) or formula (I-1) or formula (I-1B) or formula (I-1B-1) or formula (I-1A) or formula (I-1A-1) or formula (I-1A-2) or formula (I-2) or formula (I-2A) or formula (I-2B) or their stereoisomers or pharmaceutically acceptable salts or pharmaceutical compositions thereof in the prevention or treatment of GCase-mediated diseases.

[0270] On the other hand, this disclosure provides compounds of formula (I) or formula (I-1) or formula (I-1B) or formula (I-1B-1) or formula (I-1A) or formula (I-1A-1) or formula (I-1A-2) or formula (I-2) or formula (I-2A) or formula (I-2B) for the prevention or treatment of GCase-mediated diseases, or stereoisomers thereof, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions thereof.

[0271] On the other hand, this disclosure provides a method for allosteric regulation of GCase, comprising administering to a mammal, preferably a human, a therapeutically effective amount of a compound of formula (I) or formula (I-1) or formula (I-1B) or formula (I-1B-1) or formula (I-1A) or formula (I-1A-1) or formula (I-1A-2) or formula (I-2) or formula (I-2A) or formula (I-2B) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0272] On the other hand, this disclosure provides the use of a compound of formula (I) or formula (I-1) or formula (I-1B) or formula (I-1B-1) or formula (I-1A) or formula (I-1A-1) or formula (I-1A-2) or formula (I-2) or formula (I-2A) or formula (I-2B) or its stereoisomer or pharmaceutically acceptable salt or pharmaceutical composition thereof in the preparation of a medicament for allosteric regulation of GCase.

[0273] On the other hand, this disclosure provides the use of compounds of formula (I) or formula (I-1) or formula (I-1B) or formula (I-1B-1) or formula (I-1A) or formula (I-1A-1) or formula (I-1A-2) or formula (I-2) or formula (I-2A) or formula (I-2B) or their stereoisomers or pharmaceutically acceptable salts or pharmaceutical compositions thereof in allosteric modulation of GCase.

[0274] On the other hand, this disclosure provides compounds of formula (I) or (I-1) or (I-1B) or (I-1B-1) or (I-1A) or (I-1A-1) or (I-1A-2) or (I-2) or (I-2A) or (I-2B) for allosteric regulation of GCase, or stereoisomers thereof, or pharmaceutically acceptable salts thereof, or pharmaceutical compositions thereof.

[0275] In some embodiments, the GCase-mediated diseases are those that benefit from allosteric regulation by GCases. In some embodiments, the GCase-mediated diseases are those that benefit from the activation of GCases.

[0276] In some implementations, the GCase-mediated diseases are selected from neurodegenerative diseases.

[0277] In some implementations, the neurodegenerative disease is selected from Alzheimer's disease or Parkinson's disease.

[0278] Any embodiment of any aspect of the present invention can be combined with other embodiments without contradiction. Furthermore, in any embodiment of any aspect of the present invention, any technical feature can be applied to the same technical feature in other embodiments without contradiction.

[0279] Terminology Definitions and Explanations

[0280] Unless otherwise stated, the terms used in this disclosure have the following meanings: the definitions of groups and terms recorded in this disclosure, including their definitions as examples, exemplary definitions, preferred definitions, definitions recorded in tables, definitions of specific compounds in the examples, etc., can be arbitrarily combined and combined with each other. A particular term should not be considered uncertain or unclear unless specifically defined, but should be understood in accordance with its ordinary meaning in the art. When trade names appear herein, they are intended to refer to the corresponding product or its active ingredient.

[0281] In this article Indicates the connection site.

[0282] In this article Indicates a single bond or a double bond.

[0283] The term "stereoisomer" refers to isomers that are produced by different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers and diastereomers.

[0284] The compounds disclosed herein may have asymmetric atoms such as carbon, sulfur, nitrogen, and phosphorus atoms, or asymmetric double bonds, and therefore may exist in specific geometric or stereoisomeric forms. Specific geometric or stereoisomeric forms may be cis and trans isomers, E- and Z-type geometric isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)- isomers, (L)- isomers, and racemic mixtures thereof or other mixtures, such as mixtures enriched with enantiomers or diastereomers. All such isomers and mixtures thereof are within the scope of the definition of the compounds disclosed herein. Alkyl groups or other substituents may contain additional asymmetric carbon atoms, asymmetric sulfur atoms, asymmetric nitrogen atoms, or asymmetric phosphorus atoms. All such isomers involved in all substituents, and mixtures thereof, are also included within the scope of the definition of the compounds disclosed herein. The compounds containing asymmetric atoms disclosed herein can be isolated in optically active pure form or in racemic form. The optically active pure form can be separated from racemic mixtures or synthesized using chiral starting materials or chiral reagents.

[0285] The term "substitution" refers to the substitution of one or more hydrogen atoms on a specific atom by a substituent, provided that the valence state of the specific atom is normal and the resulting compound is stable. When the substituent is oxo (i.e., =O), it means that two hydrogen atoms are substituted; oxo substitution does not occur on aromatic groups.

[0286] The terms “optional” or “optionally” mean that the event or condition subsequently described may or may not occur, including both the occurrence and non-occurrence of said event or condition. For example, “optionally” substituted with a halogen means that the ethyl group can be unsubstituted (CH2CH3), monosubstituted (CH2CH2F, CH2CH2Cl, etc.), polysubstituted (CHFCH2F, CH2CHF2, CHFCH2Cl, CH2CHCl2, etc.), or fully substituted (CF2CF3, CF2CCl3, CCl2CCl3, etc.). Those skilled in the art will understand that for any group containing one or more substituents, no substitution or substitution pattern that is spatially impossible and / or cannot be synthesized is introduced.

[0287] When any variable (e.g., R) a R b When a group appears more than once in the composition or structure of a compound, its definition is independent in each case. For example, if a group is surrounded by two R... b Replaced, then each R b Each has its own independent options.

[0288] Unless otherwise specified, the linking direction of the linking groups mentioned in this article is arbitrary. For example, when ring B in the structural unit "W-ring BL" is selected from... At this time, ring B can be connected in a left-to-right direction to form the following unit "ring". Alternatively, the following units can be formed by connecting them from right to left.

[0289] When a substituent is cross-bonded to two atoms on a ring, it can bond to any atom on that ring. For example, structural units. R represents c Substitution can occur at any position on the pyrazole ring.

[0290] In this article, "key" in "W selected from key" refers to a direct-connected key.

[0291] C in this article m -C n It refers to having an integer number of carbon atoms in the range mn. For example, "C1-C 10 "" means that the group can have 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms.

[0292] The term "alkyl" refers to a compound with the general formula C1. n H 2n+1The alkyl group can be straight-chain or branched. The term "C1-C" refers to a hydrocarbon group. 10 "Alkyl" can be understood as representing a straight-chain or branched saturated hydrocarbon group having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms. Specific examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, and 2,2-dimethylbutyl. 1,1-Dimethylbutyl, 2,3-Dimethylbutyl, 1,3-Dimethylbutyl, or 1,2-Dimethylbutyl, etc.; the term "C1-C6 alkyl" can be understood as referring to alkyl groups having 1, 2, 3, 4, 5, or 6 carbon atoms, specific examples including but not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, neopentyl, hexyl, 2-methylpentyl, etc. The term "C1-C3 alkyl" can be understood as referring to straight-chain or branched saturated alkyl groups having 1, 2, or 3 carbon atoms. The "C1-C6"... 10 "alkyl" can include the range of "C1-C6 alkyl" or "C1-C3 alkyl", and the "C1-C6 alkyl" can further include "C1-C3 alkyl".

[0293] The term “halogenated alkyl” is intended to include both monohalogenated and polyhalogenated alkyl groups. For example, the term “C1-C6 haloalkyl” means a C1-C6 alkyl group as defined above that is substituted with one or more halogens, including but not limited to trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, trichloromethyl, pentafluoroethyl, and pentachloroethyl, etc.

[0294] The term "alkoxy" refers to the substitution of a hydrogen atom on the hydroxyl group of a straight-chain or branched alcohol by an alkyl group; it can be understood as "alkyloxy" or "alkyl-O-". The term "C1-C"... 10 "Alkoxy" can be understood as "C1-C" 10 "alkyloxy" or "C1-C" 10 Alkyl-O-"; the term "C1-C6 alkoxy" can be understood as "C1-C6 alkyloxy" or "C1-C6 alkyl-O-". The "C1-C" 10 "Alkoxy" can include the range of "C1-C6 alkoxy" and "C1-C3 alkoxy", and the "C1-C6 alkoxy" can further include "C1-C3 alkoxy".

[0295] The term "cycloalkyl" refers to a fully saturated carbon ring existing in the form of a monocyclic, fused, bridged, or spirocyclic ring. Unless otherwise indicated, the carbon ring is typically a 3- to 14-membered ring. The term "C3-C" is also used. 12 "Cycloalkyl" can be understood as representing a saturated monocyclic, fused, spirocyclic, or bridged ring having 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 carbon atoms. Specific examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, norbornyl (bicyclo[2.2.1]heptyl), bicyclo[2.2.2]octyl, adamantyl, spiro[4.5]decyl, etc. The term "C3-C" 12 "Cycloalkyl" can include "C3-C6 cycloalkyl". The term "C3-C6 cycloalkyl" can be understood as indicating a saturated monocyclic or bicyclic hydrocarbon ring having 3, 4, 5 or 6 carbon atoms. Specific examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

[0296] The term "spirocycloalkyl" refers to a 6- to 14-membered polycyclic group in which the monocyclic rings share a single carbon atom (called the spiro atom), which may contain one or more double bonds but does not have a fully conjugated π-electron system as a whole. Preferably, it is 7 to 12-membered (e.g., 7, 8, 9, 10, 11, or 12-membered). More preferably, it is a 4- / 4-membered, 4- / 5-membered, 4- / 6-membered, 5- / 5-membered, or 5- / 6-membered bicyclic spirocycloalkyl group. Wherein, A-membered / B-membered bicyclic spirocycloalkyl refers to a spirocycle formed by an A-membered monocyclic ring and a B-membered monocyclic ring sharing a single atom. Specific examples of spirocycloalkyl groups include, but are not limited to:

[0297] The term "spirocycloalkyl" refers to a divalent group derived from a "spirocycloalkyl" group as defined herein.

[0298] The term "fused cycloalkyl" refers to a 6- to 14-membered polycyclic aromatic hydrocarbon group in which each ring shares an adjacent pair of carbon atoms with other rings in the system. One or more rings may contain one or more double bonds, but the group does not have a fully conjugated π-electron system overall. Preferably, it is 7- to 12-membered (e.g., 7, 8, 9, 10, 11, or 12-membered). Depending on the number of rings, it can be classified as bicyclic, tricyclic, tetracyclic, or polycyclic fused cycloalkyl, preferably bicyclic or tricyclic, more preferably 5-membered / 5-membered or 5-membered / 6-membered bicyclic fused cycloalkyl. A-membered / B-membered bicyclic fused cycloalkyl refers to a group formed by an A-membered monocyclic ring and a B-membered monocyclic ring sharing an adjacent pair of atoms. Specific examples of fused cycloalkyl groups include, but are not limited to:

[0299] The term “condensed cycloalkyl” refers to a divalent group derived from a “condensed cycloalkyl” group as defined herein.

[0300] The term "bridged cycloalkyl" refers to a 5- to 12-membered, all-carbon polycyclic group in which any two rings share two non-directly bonded carbon atoms. It may contain one or more double bonds, but does not possess a fully conjugated π-electron system overall. Preferably, it is 5- to 9-membered (e.g., 5, 6, 7, 8, or 9-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. Specific examples of bridged cycloalkyl groups include, but are not limited to:

[0301] The term “bridged cycloalkyl” refers to a divalent group derived from a “bridged cycloalkyl” group as defined herein.

[0302] The term "heterocyclic group" refers to a fully saturated or partially saturated (not aromatic as a whole) monocyclic, fused-ring, spirocyclic, or bridged ring containing 1, 2, 3, 4, or 5 heteroatoms or heterogroups (i.e., groups containing heteroatoms). These "heteroatoms or heterogroups" include, but are not limited to, nitrogen (N), oxygen (O), sulfur (S), phosphorus (P), boron (B), -S(=O)2-, -S(=O)-, -P(=O)2-, -P(=O)-, -NH-, -S(=O)(=NH)-, -C(=O)NH-, or -NHC(=O)NH-. The term "4-12 membered heterocyclic group" refers to a heterocyclic group with 4, 5, 6, 7, 8, 9, 10, 11, or 12 ring atoms, containing 1, 2, 3, 4, or 5 heteroatoms or heterogroups independently selected from those described above. "3-10 membered heterocyclic groups" include "4-7 membered heterocyclic groups", wherein specific examples of 4 membered heterocyclic groups include, but are not limited to, azirrocyclobutane, thioheterocyclobutane, or oxoheterocyclobutane; specific examples of 5 membered heterocyclic groups include, but are not limited to, tetrahydrofuranyl, dioxacyclopentenyl, pyrrolyl, imidazoalkyl, pyrazolyl, pyrrolinyl, 4,5-dihydrooxazolyl, or 2,5-dihydro-1H-pyrrolyl; specific examples of 6 membered heterocyclic groups include, but are not limited to, tetrahydropyranyl, piperidinyl, morpholinyl, dithiaalkyl, thiomorpholinyl, piperazinyl, trithiaalkyl, tetrahydropyridinyl, or 4H-[1,3,4]thiadiazinyl; and specific examples of 7 membered heterocyclic groups include, but are not limited to, diazacycloheptane. The heterocyclic group can also be a bicyclic group, wherein specific examples of 5-membered / 5-membered bicyclic groups include, but are not limited to, hexahydrocyclopentano[c]pyrrolo-2(1H)-yl or; specific examples of 5-membered / 6-membered bicyclic groups include, but are not limited to, hexahydropyrrolo[1,2-a]pyrazin-2(1H)-yl, 5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazinyl or 5,6,7,8-tetrahydroimidazo[1,5-a]pyrazinyl. Optionally, the heterocyclic group can be a benzofused cyclic group of the above-mentioned 4-7-membered heterocyclic groups, specific examples of which include, but are not limited to, dihydroisoquinolinyl, etc. The term "4-10 membered heterocyclic group" can include the ranges of "5-10 membered heterocyclic group", "4-7 membered heterocyclic group", "5-6 membered heterocyclic group", "6-8 membered heterocyclic group", "4-10 membered heterocyclic alkyl group", "5-10 membered heterocyclic alkyl group", "4-7 membered heterocyclic alkyl group", "5-6 membered heterocyclic alkyl group", and "6-8 membered heterocyclic alkyl group". "4-7 membered heterocyclic group" can further include the ranges of "4-6 membered heterocyclic group", "5-6 membered heterocyclic group", "4-7 membered heterocyclic alkyl group", "4-6 membered heterocyclic alkyl group", and "5-6 membered heterocyclic alkyl group". Although some bicyclic heterocyclic groups in this disclosure partially contain a benzene ring or a heteroaromatic ring, the heterocyclic group as a whole is still non-aromatic.

[0303] The term "spiroheterocyclic group" refers to a polycyclic heterocyclic group in which 6- to 14-membered monocyclic rings share a single atom (called a spiro atom), wherein one or more ring atoms are heteroatoms selected from nitrogen (N), oxygen (O), sulfur (S), S(O), or S(O)2, and the remaining ring atoms are carbon. It may contain one or more double bonds, but does not possess a fully conjugated π-electron system overall. Preferably, it is 7 to 12-membered (e.g., 7, 8, 9, 10, 11, or 12-membered). More preferably, it is a 4-membered / 4-membered, 4-membered / 5-membered, 4-membered / 6-membered, 5-membered / 5-membered, or 5-membered / 6-membered bicyclic spiroheterocyclic group. Specifically, an A-membered / B-membered bicyclic spiroheterocyclic group refers to a spirocyclic group formed by an A-membered monocyclic ring and a B-membered monocyclic ring sharing a single atom. Specific examples of spiroheterocyclic groups include, but are not limited to:

[0304] The term "spiroheterocyclic group" refers to a divalent group derived from a "spiroheterocyclic group" as defined herein.

[0305] The term "fused heterocyclic group" refers to a 6- to 14-membered polycyclic heterocyclic group in which each ring in the system shares an adjacent pair of atoms with other rings in the system. One or more rings may contain one or more double bonds, but do not have a fully conjugated π-electron system overall. One or more ring atoms are heteroatoms selected from nitrogen (N), oxygen (O), sulfur (S), S(O), or S(O)2, and the remaining ring atoms are carbon. Preferably, it is 7- to 12-membered (e.g., 7, 8, 9, 10, 11, or 12-membered). Depending on the number of rings, it can be classified as bicyclic, tricyclic, tetracyclic, or polycyclic fused heterocyclic groups, preferably bicyclic or tricyclic, more preferably 3-membered / 5-membered, 5-membered / 5-membered, or 5-membered / 6-membered bicyclic fused heterocyclic groups. A-membered / B-membered bicyclic fused heterocyclic groups refer to groups formed by an A-membered monocyclic ring and a B-membered monocyclic ring sharing an adjacent pair of atoms. Specific examples of fused heterocyclic groups include, but are not limited to:

[0306] The term “fused heterocyclic group” refers to a divalent group derived from a “fused heterocyclic group” as defined herein.

[0307] The term "bridged heterocyclic group" refers to a 5- to 12-membered polycyclic heterocyclic group in which any two rings share two non-directly bonded atoms. It may contain one or more double bonds, but does not possess a fully conjugated π-electron system overall. One or more ring atoms are heteroatoms selected from nitrogen (N), oxygen (O), S(O), or S(O)₂, and the remaining ring atoms are carbon. Preferably, it is 5- to 9-membered (e.g., 5, 6, 7, 8, or 9-membered). Depending on the number of rings, it can be classified as bicyclic, tricyclic, tetracyclic, or polycyclic bridged heterocyclic groups, preferably bicyclic, tricyclic, or tetracyclic, and more preferably bicyclic or tricyclic. Specific examples of bridged heterocyclic groups include, but are not limited to:

[0308] The term “bridged heterocyclic group” refers to a divalent group derived from a “bridged heterocyclic group” as defined herein.

[0309] The term "aryl" refers to an aromatic ring group consisting of an all-carbon monocyclic or fused polycyclic aromatic cyclic group with a conjugated π-electron system. Aryl groups can have 6-20, 6-14, or 6-12 carbon atoms. The term "C6-C"... 20 "Aryl" can be understood as an aryl group having 6 to 20 carbon atoms. Specifically, it refers to a ring with 6 carbon atoms ("C6 aryl"), such as phenyl; or a ring with 9 carbon atoms ("C9 aryl"), such as indenyl or indenyl; or a ring with 10 carbon atoms ("C9 aryl"). 10 Aryl groups, such as tetrahydronaphthyl, dihydronaphthyl, or naphthyl; or rings having 13 carbon atoms (“C”). 13 Aryl groups, such as fluorene groups; or rings with 14 carbon atoms (“C”). 14 Aryl), for example, anthracene. The term "C6-C" is used. 10 "Aryl" can be understood as an aryl group having 6 to 10 carbon atoms. Specifically, it refers to a ring with 6 carbon atoms ("C6 aryl"), such as phenyl; or a ring with 9 carbon atoms ("C9 aryl"), such as indenyl or indenyl; or a ring with 10 carbon atoms ("C9 aryl"). 10 Aryl), such as tetrahydronaphthyl, dihydronaphthyl, or naphthyl. The term "C6-C" is used. 20 "Aryl" can contain "C6-C 10 Aryl.

[0310] The term "heteroaryl" refers to an aromatic monocyclic or fused polycyclic system containing at least one ring atom selected from N, O, or S, with the remaining ring atoms being C. The term "5-10-membered heteroaryl" can be understood to include monocyclic or bicyclic aromatic ring systems having 5, 6, 7, 8, 9, or 10 ring atoms, particularly 5, 6, 9, or 10 ring atoms, and containing 1, 2, 3, 4, or 5, preferably 1, 2, or 3 heteroatoms independently selected from N, O, and S. Specifically, the heteroaryl group is selected from thienyl, furanyl, pyrroleyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl or thiadiazolyl and their benzo[derivatives], such as benzofuranyl, benzothienyl, benzothiazolyl, benzooxazolyl, benzoisooxazolyl, benzoimidazolyl, benzotriazolyl, indazole, indolyl or isindolyl; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl or triazinyl and their benzo[derivatives], such as quinolinyl, quinazolinyl or isoquinolinyl; or acrylinyl, inazinyl, purinyl and their benzo[derivatives]; or cyclolinyl, phthalazinyl, quinazolinyl, quinoxolinyl, naphthidyl, pteridinyl, carbazolyl, acrylinyl, phenazinyl, phenothiazinyl or phenothiazinyl. The term "5-6 heteroaryl" refers to an aromatic ring system having 5 or 6 ring atoms, and containing 1, 2 or 3, preferably 1-2 heteroatoms independently selected from N, O and S.

[0311] The term "halogen" or "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0312] The term "treatment" means administering the compound or preparation described in this application to improve or eliminate a disease or one or more symptoms related to said disease, and includes:

[0313] (i) Suppress the disease or disease state, that is, curb its development;

[0314] (ii) Relieve the disease or disease state, even if the disease or disease state subsides.

[0315] The term “prevention” means administering the compound or formulation described in this application to prevent a disease or one or more symptoms associated with the disease, including preventing the occurrence of a disease or disease state in mammals, particularly when such mammals are susceptible to the disease state but have not yet been diagnosed with the disease state.

[0316] The term "therapeutic effective dose" means:

[0317] The amount of the disclosed compound used to treat a specific disease, condition or symptom, (ii) reduce, improve or eliminate one or more symptoms of a specific disease, condition or symptom, or (iii) delay the onset of one or more symptoms of a specific disease, condition or symptom described herein.

[0318] The amount of the disclosed compound constituting a “therapeutic effective amount” varies depending on the compound, the disease state and its severity, the route of administration, and the age of the mammal to be treated, but may routinely be determined by a person skilled in the art based on their own knowledge and the content of this disclosure.

[0319] Examples of the term "mammal" include, but are not limited to, any member of the class Mammalia: humans, non-human primates (e.g., chimpanzees and other apes and monkeys); livestock, such as cattle, horses, sheep, goats, and pigs; domesticated animals, such as rabbits, dogs, and cats; and laboratory animals, including rodents, such as rats, mice, and guinea pigs. In one embodiment of the methods and compositions provided herein, the mammal may be a human.

[0320] The term "pharmaceutical acceptable" refers to compounds, materials, compositions, and / or dosage forms that, within the bounds of reliable medical judgment, are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0321] The term "pharmaceutically acceptable salt" refers to a pharmaceutically acceptable salt that has undergone acid or base addition, including salts formed by compounds with inorganic or organic acids, and salts formed by compounds with inorganic or organic bases.

[0322] The term "pharmaceutical composition" refers to a mixture of one or more compounds of the present disclosure or salts thereof with pharmaceutically acceptable excipients. The purpose of a pharmaceutical composition is to facilitate the administration of the disclosed compounds to an organism.

[0323] The term "pharmaceuticalally acceptable excipient" refers to excipients that do not cause significant irritation to the organism and do not impair the biological activity and properties of the active compound. Suitable excipients are well known to those skilled in the art, such as carbohydrates, waxes, water-soluble and / or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, etc.

[0324] The word “comprise” or “include” and its English variants such as comprises or comprising can be understood as having an open, non-exclusive meaning, that is, “including but not limited to”.

[0325] This disclosure also includes compounds of this disclosure that are identical to those described herein, but in which one or more atoms are labeled with isotopes whose atomic weights or mass numbers differ from those commonly found in nature. Examples of isotopes that can be incorporated into compounds of this disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine, and chlorine, such as... 2H, 3 H, 11 C 13 C 14 C 13 N、 15 N、 15 O、 17 O、 18 O、 31 P, 32 P, 35 S, 18 F, 123 I, 125 I and 36 Cl, etc.

[0326] Certain isotope-labeled compounds of this disclosure (e.g., using...) 3 H and 14 C-labeling can be used in the analysis of compound and / or substrate tissue distribution. Tritiumization (i.e., 3 H) and carbon-14 (i.e. 14 C) Isotopes are particularly preferred due to their ease of preparation and detectability. Positron-emitting isotopes, such as... 15 O、 13 N、 11 C and 18 F can be used in positron emission tomography (PET) studies to determine substrate occupancy. The isotopically labeled compounds of this disclosure can typically be prepared by replacing the unlabeled reagent with an isotopically labeled reagent using a procedure similar to those disclosed in the schemes and / or examples below.

[0327] The pharmaceutical compositions disclosed herein can be prepared by combining the compounds disclosed herein with suitable pharmaceutically acceptable excipients, for example, in solid, semi-solid, liquid or gaseous formulations, such as tablets, pills, capsules, powders, granules, ointments, emulsions, suspensions, suppositories, injections, inhalers, gels, microspheres and aerosols.

[0328] Typical routes of administration of the disclosed compounds or their pharmaceutically acceptable salts or pharmaceutical compositions thereof include, but are not limited to, oral, rectal, topical, inhalation, parenteral, sublingual, vaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, and intravenous administration.

[0329] The pharmaceutical compositions disclosed herein can be manufactured using methods well known in the art, such as conventional mixing, dissolving, granulation, emulsification, freeze drying, etc.

[0330] In some embodiments, the pharmaceutical composition is in an oral form. For oral administration, the pharmaceutical composition can be formulated by mixing the active compound with pharmaceutically acceptable excipients well known in the art. These excipients enable the compounds of this disclosure to be formulated into tablets, pills, lozenges, sugar-coated tablets, capsules, liquids, gels, pastes, suspensions, etc., for oral administration to patients.

[0331] Solid oral compositions can be prepared using conventional mixing, filling, or tableting methods. For example, they can be obtained by mixing the active compound with solid excipients, optionally milling the resulting mixture, adding other suitable excipients if necessary, and then processing the mixture into granules to obtain the core of a tablet or sugar-coated formulation. Suitable excipients include, but are not limited to, binders, diluents, disintegrants, lubricants, flow aids, or flavoring agents.

[0332] The pharmaceutical composition may also be suitable for parenteral administration, such as in suitable unit dosage forms of sterile solutions, suspensions or lyophilized products.

[0333] In all methods of administration of the compound of general formula (I) described herein, or its stereoisomers or pharmaceutically acceptable salts thereof, the daily dose is from 0.01 mg / kg to 1000 mg / kg body weight, in single or separate doses. Detailed Implementation

[0334] The invention will now be described in detail with reference to embodiments, but this does not imply any adverse limitation of the present disclosure. The present disclosure has been described in detail herein, including specific embodiments thereof. It will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of the present disclosure without departing from the spirit and scope thereof.

[0335] Unless otherwise specified, all reagents used in this disclosure are commercially available and can be used without further purification.

[0336] Unless otherwise stated, the proportions expressed for mixed solvents are volume-based.

[0337] Unless otherwise stated, % refers to wt%.

[0338] Compounds are processed manually or Software naming conventions are used; commercially available compounds use supplier catalog names.

[0339] The structure of the compound was determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). NMR shifts are measured in units of 10⁻⁶. -6 (ppm). The solvents used for NMR determination included deuterated dimethyl sulfoxide, deuterated chloroform, and deuterated methanol, with tetramethylsilane (TMS) as the internal standard. "IC50 "Half-inhibitory concentration" refers to the concentration at which half of the maximum inhibitory effect is achieved.

[0340] The eluent described below can be a mixture of two or more solvents, with the ratio being the volume ratio of each solvent. For example, in a mixed eluent, dichloromethane / methanol:10 / 1 indicates that the volume ratio of dichloromethane to methanol is 10:1.

[0341] Abbreviations:

[0342] THF: Tetrahydrofuran; DBU: 1,8-diazabicyclo[5.4.0]undec-7-ene; EtOH: Ethanol; CDI: N,N'-carbonyldiimidazole; DMF: N,N-dimethylformamide; MeI: Iodomethane; DMA: N,N-dimethylaniline; TEA: Triethylamine; HATU: O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethylurea hexafluorophosphate; DCM: Dichloromethane; TFA: Trifluoroacetic acid; mCPBA: m-chloroperoxybenzoic acid; Ac2O Acetic anhydride; Pd2(dba)3: tris(dibenzylacetone)dipalladium; Xantphos: 4,5-bis(diphenylphosphine-9,9-dimethyloxanthracene; Dioxane: 1,4-dioxane; MeOH: methanol; TOSMIC: p-toluenesulfonylmethylisocyanate; t-BuOK: potassium tert-butoxide; DME: ethylene glycol dimethyl ether; Rf: ratio of solute migration distance to mobile phase migration distance; TFAA: trifluoroacetic anhydride; TMSCl: trimethylchlorosilane; ACN: acetonitrile; RockPhosPd G3: Methanesulfonate-(2-(di-tert-butylphosphine)-3-methoxy-6-methyl,2,4,6-triisopropyl-1,1-biphenyl)(2-amino-1,1-biphenyl-2-yl)palladium(II); DIEA or DIPEA: N,N-diisopropylethylamine; KHDMS: bis(trimethylsilyl)aminopotassium; Pd(dppf)Cl2: 1,1-bis(diphenylphosphine)ferrocene palladium dichloride; KOAc: potassium acetate; B2Pin2: pinacolyl diboronate; MeOH: methanol; MsCl: methanesulfonyl chloride; EA: ethyl acetate; Xphos Pd G4: (SP-4-3)-[dicyclohexyl[2',4',6'-tris(isopropyl)[1,1'-biphenyl]-2-yl]phosphine](methanesulfonic acid)[2'-(methylamino)[1,1'-biphenyl]-2-yl]palladium; Tf represents trifluoromethanesulfonyl; TBAF: tetrabutylammonium fluoride.

[0343] Example 1: 1-(2,2-difluoroethyl)-4-methyl-6-(6-(2-(trifluoromethyl)pyridin-4-yl)-6-azaspiro[3,4]octane-2-yl)-1,4-dihydro-5H-pyrazolo[4,3-d]pyrimidine-5,7(6H)-dione

[0344] Step 1: Synthesis of intermediates 1-2

[0345] Compound 1-1 (250 mg) and methyl 1-(2,2-difluoroethyl)-4-nitro-1H-pyrazole-3-carboxylic acid (390 mg) were dissolved in 5 mL of tetrahydrofuran. Then, 1,8-diazabicyclo[5.4.0]undec-7-ene (336 mg) was added at 0 °C. The reaction mixture was stirred overnight at 60 °C until complete. After cooling to room temperature, 50 mL of water was added, followed by extraction with ethyl acetate (20 mL * 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (petroleum ether / ethyl acetate: 2 / 1; Rf: 0.6) to obtain compound 1-2 (400 mg).

[0346] Step 2: Synthesis of intermediates 1-3

[0347] Compounds 1-2 (390 mg) were dissolved in an ethanol / water mixture (5 mL; 4:1), followed by the addition of iron powder (253 mg) and ammonium chloride (486 mg). The reaction mixture was stirred at 80 °C for 2 hours until the reaction was complete. After cooling to room temperature, the reaction mixture was filtered. The filtrate was added to 30 mL of water and extracted with ethyl acetate (20 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compounds 1-3 (350 mg).

[0348] Step 3: Synthesis of intermediates 1-4

[0349] Compounds 1-3 (280 mg) were dissolved in 5 mL of tetrahydrofuran, and N,N'-carbonyldiimidazole (341 mg) was added. The reaction mixture was stirred at 70 °C for 2 hours until the reaction was complete. The reaction mixture was cooled to room temperature, 10 mL of water was added, and then extracted with ethyl acetate (20 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol: 10 / 1) to obtain the title compound 1-4 (240 mg).

[0350] Step 4: Synthesis of compounds 1-5

[0351] Compounds 1-4 (220 mg) and potassium carbonate (214 mg) were dissolved in N,N-dimethylformamide (3 mL), and iodomethane (73.4 mg) was added dropwise. The reaction mixture was stirred at 60 °C for 1 hour until the reaction was complete. The reaction mixture was cooled to room temperature, 10 mL of water was added, and then extracted with ethyl acetate (30 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol: 10 / 1) to obtain the title compound 1-5 (160 mg).

[0352] Step 5: Synthesis of intermediates 1-6

[0353] Under nitrogen protection, compounds 1-5 (150 mg) were added to a 4 M ethyl hydrochloride solution (10 mL). The reaction was carried out at room temperature for 2 hours until complete. The reaction solution was concentrated under reduced pressure to obtain the title compound 1-6 (120 mg).

[0354] MS m / z (ESI): 340.2 [M+H] +

[0355] Step 6: Synthesis of Compound 1

[0356] Compounds 1-6 (100 mg) and 4-bromo-2-(trifluoromethyl)pyridine (90.2 mg) were dissolved in N,N-dimethylformamide (3 mL), and potassium carbonate (110 mg) was added. The reaction was carried out after heating to 80 °C and stirring for 2 hours. The reaction solution was cooled to room temperature, 10 mL of water was added, and then extracted with ethyl acetate (20 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol: 10 / 1) and preparative high performance liquid chromatography (column: Phenomenex Gemini NX 150 × 30 mm, 5 μm; mobile phase: [water (ammonium bicarbonate, 7 mmol / L) - acetonitrile]; acetonitrile gradient: 50%-90%) to obtain compound 1 (35.0 mg).

[0357] 1 H NMR (400MHz, DMSO-d6) δ8.20 (d, J = 5.8Hz, 1H), 7.90 (s, 1H), 6.96-6.76 (m, 1H), 6.72-6.59 (m, 1H), 6.60-6.18 (m, 1H), 5.56-5.30(m,1H),5.16-4.80(m,2H),3.52-3.35(m,7H),3.11-2.92(m,2H),2.33-2.20(m,2H),2.13(t,J=6.7Hz,2H).

[0358] MS m / z (ESI): 485.2 [M+H] +

[0359] Examples 2-4

[0360] The synthesis method of Example 1 was used, except that 1-1 in step 1 was replaced with the starting materials in the table below, and the following compounds 2-4 were synthesized in the same way.

[0361] Example 5: 7-Methyl-3-(6-(2-(trifluoromethyl)pyridin-4-yl)-6-azaspiro[3.4]oct-2-yl)pyrimido[4,5-d]pyrimidin-4(3H)-one

[0362] Step 1: Synthesis of intermediate 5-2

[0363] 2-Methyl-4-aminopyrimidine-5-carboxylic acid (200 mg) was dissolved in N,N-dimethylaniline (4 mL), followed by the addition of triethylamine (122 mg) and O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethylurea hexafluorophosphate (462 mg). The mixture was stirred at room temperature for 1 hour. Compound 1-1 (250 mg) was then added, and the reaction was continued for 2 hours until completion. 50 mL of water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (20 mL x 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by column chromatography (petroleum ether / ethyl acetate: 2 / 1; Rf: 0.6) to obtain the title compound 5-2.

[0364] Step 2: Synthesis of intermediate 5-3

[0365] Compound 5-2 (400 mg) was dissolved in 5 mL of formamide, heated to 120 °C, and stirred for 40 hours until the reaction was complete. The reaction solution was cooled to room temperature, 50 mL of water was added, and the mixture was extracted with ethyl acetate (20 mL * 3). The organic phase was washed with saturated brine and dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by column chromatography (petroleum ether / ethyl acetate: 1 / 1; Rf: 0.5) to obtain the title compound 5-3 (240 mg).

[0366] Step 3: Synthesis of intermediate 5-4

[0367] Compound 5-3 (240 mg) was dissolved in 6 mL of dichloromethane, and 2 mL of trifluoroacetic acid was slowly added dropwise at 0 °C. The reaction was carried out after stirring at room temperature for 2 hours. The reaction solution was then concentrated under reduced pressure to obtain the title compound 5-4 (240 mg).

[0368] Step 4: Synthesis of Compound 5

[0369] 4-Bromo-2-(trifluoromethyl)pyridine (136 mg), compound 5-4 (240 mg), and potassium carbonate (250 mg) were dissolved in N,N-dimethylformamide (3.00 mL), and the mixture was heated to 80 °C and stirred for 6 hours until the reaction was complete. 50 mL of water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (20 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by high performance liquid chromatography (column: YMC TAR-C18, 30 * 150 mm, 5 μm; mobile phase: [water (containing 7 mmol / L ammonium bicarbonate) - acetonitrile]; acetonitrile gradient: 40-70%) to obtain title compound 5 (115 mg).

[0370] 1 H NMR (400MHz, DMSO-d6) δ9.40 (s, 1H), 8.86 (d, J = 2.8Hz, 1H), 8.22 (d, J = 5.8Hz, 1H), 6.95-6.56 (m ,2H),5.13-4.97(m,1H),3.63-3.35(m,4H),2.75(s,3H),2.68-2.56(m,4H),2.24-2.05(m,2H).

[0371] MS m / z (ESI): 417.2 [M+H] +

[0372] Example 6: 1-(2,2-difluoroethyl)-4-methyl-6-((3aR,6aS)-5-(2-(trifluoromethyl)pyridin-4-yl)hexahydropyrrolo[3,4-c]pyrrolo-2(1H)-yl)-1,4-dihydro-5H-pyrazolo[4,3-b]pyridin-5-one

[0373] Step 1: Synthesis of intermediate 6-2:

[0374] 6-Bromo-1H-pyrazolo[4,3-b]pyridine (1.00 g) and cesium carbonate (3.29 g) were dissolved in N,N-dimethylformamide (15 mL). 2,2-Difluoroethyltrifluoromethanesulfonate (1.19 g) was added dropwise at 0 °C. After the addition was complete, the mixture was stirred at room temperature for 2 hours until the reaction was complete. The reaction solution was cooled to room temperature, 100 mL of water was added, and the mixture was extracted with ethyl acetate (100 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (petroleum ether / ethyl acetate: 2 / 1) to obtain the title compound 6-2 (0.57 g).

[0375] 1H NMR (400MHz, DMSO-d6) δ8.74-8.61(m,2H),8.46(d,J=1.0Hz,1H),6.69-6.28(m,1H),5.13-4.90(m,2H).

[0376] Step 2: Synthesis of intermediate 6-3:

[0377] Under nitrogen protection, compound 6-2 (420 mg) was dissolved in 10 mL of dichloromethane, and m-chloroperoxybenzoic acid (390 mg) was added at 0 °C. The reaction was stirred at room temperature for 16 hours until completion. The reaction solution was quenched with 30 mL of saturated sodium bisulfite, then extracted with ethyl acetate (30 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (petroleum ether / ethyl acetate: 1 / 1) to obtain the title compound 6-3 (400 mg).

[0378] 1 H NMR (400MHz, DMSO-d6) δ8.53(d,J=1.0Hz,1H),8.47(d,J=0.9Hz,1H),8.27(s,1H),6.63-6.30(m,1H),5.10-4.90(m,2H).

[0379] Step 3: Synthesis of intermediate 6-4:

[0380] Under nitrogen protection, compound 6-3 (400 mg) was added to acetic anhydride (20 mL). The mixture was heated to reflux and stirred for 2 hours. The reaction mixture was then cooled to 90 °C, 10 mL of water was added, and the mixture was stirred at 90 °C for 2 hours until the reaction was complete. The reaction mixture was cooled to room temperature, the pH was adjusted to 7–8 with 2 M sodium hydroxide solution, and the mixture was extracted with ethyl acetate (30 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol: 10 / 1) to obtain the title compound 6-4 (390 mg).

[0381] m / z:ES + 277.8 [M+H] +

[0382] Step 4: Synthesis of intermediates 6-5:

[0383] Under nitrogen protection, compound 6-4 (390 mg) and potassium carbonate (387 mg) were added to N,N-dimethylformamide (10 mL), and iodomethane (298 mg) was added at 0 °C. The mixture was then heated to 60 °C and stirred for 2 hours until the reaction was complete. The reaction solution was cooled to room temperature, 50 mL of water was added, and the mixture was extracted with ethyl acetate (50.00 mL * 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol: 10 / 1) to obtain the title compound 6-5 (390 mg).

[0384] 1 H NMR (400MHz, DMSO-d6) δ8.65(s,1H),7.93(s,1H),6.57-6.22(m,1H),5.00-4.82(m,2H),3.58(s,3H).

[0385] MS m / z(ESI): 291.8 [M+H] +

[0386] Step 5: Synthesis of intermediate 6-6:

[0387] Under a nitrogen atmosphere, compound 6-5 (350 mg), cesium carbonate (1.17 g), cis-tert-butylhexahydropyrrolo[3,4-c]pyrrole-2(1H)-carboxylate (447 mg), tris(dibenzylacetone)dipalladium (109 mg), and 4,5-bisdiphenylphosphine-9,9-dimethyloxanthracene (138 mg) were added to 8 mL of 1,4-dioxane. The mixture was heated to 90 °C and stirred for 2 hours until the reaction was complete. The reaction solution was cooled to room temperature, 20 mL of water was added, and the mixture was extracted with ethyl acetate (30.00 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol: 10 / 1) to obtain the title compound 6-6 (290 mg).

[0388] Step 6: Synthesis of intermediates 6-7:

[0389] Compound 6-6 (260 mg) was added to 10 mL of 4 M ethyl hydrochloride solution, and the reaction was carried out by stirring at 25 °C for 2 hours. The reaction solution was concentrated under reduced pressure, and the residue was slurried with ethyl acetate to obtain the title compound 6-7 (220 mg).

[0390] Step 7: Synthesis of Compound 6

[0391] Compounds 6-7 (100 mg) and 4-bromo-2-(trifluoromethyl)pyridine (94.2 mg) were dissolved in N,N-dimethylformamide (3 mL), and potassium carbonate (115 mg) was added. The reaction mixture was stirred at 80 °C for 2 hours until the reaction was complete. The reaction mixture was cooled to room temperature, 10 mL of water was added, and the mixture was extracted with ethyl acetate (20 mL * 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol: 10 / 1) and preparative high performance liquid chromatography (column: Phenomenex Gemini NX 150 × 30 mm, 5 μm; mobile phase: [water (containing 7 mmol / L ammonium bicarbonate) - acetonitrile]; acetonitrile gradient: 50%-90%) to obtain title compound 6 (36.8 mg).

[0392] 1 H NMR (400MHz, DMSO-d6) δ8.23(d,J=5.8Hz,1H),7.68(s,1H),6.86(d,J=2.4Hz,1H),6.81(s,1H),6.78-6.64(m,1H),6.34(tt,J =54.9,3.8Hz,1H),4.74(td,J=15.0,3.8Hz,2H),3.73-3.60(m,4H),3.55-3.45(m,5H),3.38-3.30(m,2H),3.21-3.07(m,2H).

[0393] MS m / z (ESI): 469.1 [M+H] +

[0394] Examples 7-10:

[0395] The synthesis method of Example 6 was used, except that cis-tert-butylhexahydropyrrolo[3,4-c]pyrrole-2(1H)-carboxylic acid tert-butyl ester in step 5 was replaced with the starting material in the table below, and the following compounds 7-10 were synthesized in the same way.

[0396] Example 11: 1-(2,2-difluoroethyl)-6-((2-(2-(trifluoromethyl)pyridin-4-yl)-2-azaspiro[4.5]decane-8-yl)methyl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0397] Step 1: Synthesis of intermediate 11-2:

[0398] Compound 11-1 (1.00 g) was dissolved in 10 mL of methanol, and a 1,4-dioxane solution of hydrogen chloride (4 M, 9.87 mL) was added. The mixture was stirred at room temperature for 2 hours until the reaction was complete. The solvent was removed under reduced pressure to obtain the title compound 11-2 (740 mg), which was used directly in the next reaction without further purification.

[0399] m / z:ES + [M+H] + =154.2.

[0400] Step 2: Synthesis of intermediate 11-3:

[0401] Compound 11-2 (450 mg), 4-bromo-2-(trifluoromethyl)pyridine (1.07 g), and potassium carbonate (984 mg) were dissolved in N,N-dimethylformamide (10 mL). The mixture was heated to 80 °C and stirred for 2 hours until the reaction was complete. 10 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (20 mL * 2). The organic phase was washed three times with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (ethyl acetate / petroleum ether, ethyl acetate gradient: 5%-20%) to obtain the title compound 11-3 (550 mg).

[0402] m / z:ES + [M+H] + =299.2.

[0403] 1 H NMR(400MHz,Chloroform-d)δ8.46-8.17(m,1H),6.89-6.60(m,1H),6.57-6.41(m,1H),3 .57-3.48(m,2H),3.36(s,2H),2.51-2.38(m,4H),2.16-2.07(m,2H),2.04-1.89(m,4H).

[0404] Step 3: Synthesis of intermediate 11-4:

[0405] Compound 11-3 (270 mg) and p-toluenesulfonylmethylisocyanate (265 mg) were dissolved in a mixture of 10 mL of ethylene glycol dimethyl ether and 0.5 mL of ethanol. The mixture was stirred at 0 °C while potassium tert-butoxide (254 mg) was slowly added. The reaction mixture was then stirred at room temperature for 2 hours until the reaction was complete. The reaction was quenched with 5 mL of water, extracted with ethyl acetate (20 mL x 3), and the organic phases were combined and dried over anhydrous sodium sulfate. The organic phase was concentrated under reduced pressure and purified by column chromatography (ethyl acetate / petroleum ether, ethyl acetate gradient: 20%-40%) to give the title compound 11-4 (150 mg).

[0406] m / z:ES + [M+H] + =310.1.

[0407] 1 H NMR(400MHz,Chloroform-d)δ8.35-8.05(m,1H),6.87-6.56(m,1H),6.55-6.35(m,1H),3.5 5-3.35(m,2H),3.25-3.14(m,2H),2.78-2.68(m,1H),2.07-1.68(m,8H),1.65-1.50(m,2H).

[0408] Step 4: Synthesis of intermediate 11-5:

[0409] Compound 11-4 (90.0 mg) and nickel chloride (57.0 mg) were dissolved in 4 mL of methanol. The mixture was stirred at 0 °C while sodium borohydride (110 mg) was slowly added. The reaction was completed after stirring at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by column chromatography (methanol / dichloromethane (containing 5% triethylamine): methanol gradient: 0%-30%) to obtain the target compound 11-5 (61.0 mg).

[0410] 1 H NMR(400MHz,Chloroform-d)δ8.29-8.22(m,1H),6.85-6.63(m,1H),6.54-6.38(m,1H),3.46-3.34(m,2H),3.2 6-3.03(m,2H),2.80-2.57(m,4H),1.98-1.81(m,2H),1.84-1.62(m,4H),1.49-1.37(m,3H),1.22-0.98(m,2H).

[0411] Step 5: Synthesis of intermediate 11-6:

[0412] Compound 11-5 (50.0 mg), methyl 1-(2,2-difluoroethyl)-4-nitro-1H-pyrazole-5-carboxylic acid (57.0 mg), and 1,8-diazabicyclo[5.4.0]undec-7-ene (98.0 mg) were dissolved in 2 mL of tetrahydrofuran. The mixture was heated to 60 °C and stirred for 2 hours until the reaction was complete. 4 mL of water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (5 mL x 3). The organic phases were combined and dried over anhydrous sodium sulfate. The organic phase was concentrated under reduced pressure and purified by column chromatography (ethyl acetate-petroleum ether, ethyl acetate gradient: 20%-50%) to obtain the title compound 11-6 (70.0 mg).

[0413] m / z:ES +[M+H] + =517.3.

[0414] 1 H NMR(400MHz,Chloroform-d)δ8.29-8.22(m,1H),8.23-8.09(m,2H),6.76-6.66(m,1H),6.52-6.39(m,1H),6.40-6.04(m ,1H),5.04-4.83(m,2H),3.47-3.39(m,4H),3.32-3.06(m,2H),2.02-1.77(m,5H),1.59-1.40(m,2H),1.26-1.12(m,4H).

[0415] Step 6: Synthesis of intermediates 11-7:

[0416] Compound 11-6 (65.0 mg), iron powder (36.0 mg), and ammonium chloride (68.0 mg) were dissolved in a mixture of 2.5 mL ethanol and 0.5 mL water. The mixture was heated to 80 °C and stirred for 2 hours until the reaction was complete. The reaction solution was concentrated under reduced pressure, 10 mL of ethyl acetate was added, and the solution was washed twice with saturated brine. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the title compound 11-7 (60.0 mg), which was directly used in the next reaction.

[0417] m / z:ES + [M+H] + =487.2.

[0418] Step 7: Synthesis of Compound 11

[0419] Compound 11-7 (60.0 mg) was dissolved in 1 mL of formamide, heated to 150 °C, and stirred for 2 hours until the reaction was complete. The reaction solution was cooled to room temperature, 10 mL of water was added, and then extracted with ethyl acetate (10 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by preparative liquid chromatography (Boston Prime C18 column: 5 μm silica, 30 mm diameter, 150 mm length; using a mixture of water (containing 7 mmol / L ammonium bicarbonate) and acetonitrile in decreasing polarity (45%-75%) as eluent) to obtain title compound 11 (15.0 mg).

[0420] 1H NMR(400MHz,Chloroform-d)δ8.34-8.24(m,1H),7.99(s,1H),7.83(s,1H),6.77-6.63(m,1H),6.51-6.39(m,1H),6.38-6.01(m,1H),5.21-4.9 4(m,2H),3.99-3.86(m,2H),3.48-3.35(m,2H),3.26-3.04(m,2H),2.01 -1.82(m,2H),1.77-1.67(m,5H),1.50-1.36(m,2H),1.34-1.17(m,2H).

[0421] MS m / z(ESI): 497.1 [M+H] +

[0422] Example 12: 1-(2,2-difluoroethyl)-6-((6-(2-(trifluoromethyl)pyridin-4-yl)-6-azaspiro[3,4]octane-2-yl)methyl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0423] Referring to the synthesis method of Example 11, compound 11-1 in step 1 was replaced with compound 12-1. The title compound 12 was prepared by the same method.

[0424] 1 H NMR(400MHz,Chloroform-d)δ8.38-8.22(m,1H),7.98(d,J=1.3Hz,1H),7.84(d,J=2.0Hz,1H),6.71-6.66(m,1H),6.48-6.38(m,1 H),6.20(tt,J=55.4,4.4Hz,1H),5.12-4.94(m,2H),4.36-3.96(m,2H),3.47-3.24(m,4H),2.91-2.74(m,1H),2.40-1.92(m,6H).

[0425] MS m / z(ESI): 469.0 [M+H] +

[0426] Example 13: 1-(2,2-difluoroethyl)-6-((7-(2-(trifluoromethyl)pyridin-4-yl)-7-azaspiro[3.5]nonane-2-yl)methyl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0427] Referring to the synthesis method of Example 11, compound 11-1 in step 1 was replaced with compound 13-1. The title compound 13 was prepared by the same method.

[0428] 1 H NMR(400MHz,Chloroform-d)δ8.30(d,J=6.0Hz,1H),7.98(s,1H),7.85(s,1H),6.99(d,J=2.6Hz,1H),6.76-6.70(m,1H),6.21(tt,J=5 5.5, 4.4Hz, 1H), 5.09-4.96 (m, 2H), 4.10 (d, J = 7.4Hz, 2H), 3.41-3.27 (m, 4H), 2.86-2.70 (m, 1H), 2.11-2.01 (m, 2H), 1.81-1.57 (m, 6H).

[0429] MS m / z (ESI): 483.2 [M+H] +

[0430] Example 14: 1-(2,2-difluoroethyl)-6-((2-(2-(trifluoromethyl)pyridin-4-yl)-2-azaspiro[3,4]octane-6-yl)methyl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0431] Referring to the synthesis method of Example 11, compound 11-1 in step 1 was replaced with compound 14-1. The title compound 14 was prepared by the same method.

[0432] 1 H NMR(400MHz,Chloroform-d)δ7.99(s,1H),7.87(s,1H),7.84(d,J=2.7Hz,1H),7.45(d,J=8.5Hz,1H),6.75-6.66(m,1H),6.21(tt,J=55.5,4.4Hz,1H), 5.03(td,J=13.1,4.4Hz,2H),4.11-3.96(m,2H),3.93(s,2H),3.81(s,2H), 2.66-2.42(m,1H),2.32-1.78(m,4H),1.75-1.63(m,1H),1.58-1.39(m,1H).

[0433] MS m / z (ESI): 469.2 [M+H] +

[0434] Example 15: 1-(2,2-difluoroethyl)-6-(((3aR,6aS)-2-(2-(trifluoromethyl)pyridin-4-yl)octahydrocyclopentan[c]pyrrolo-5-yl)methyl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0435] Referring to the synthesis method of Example 11, compound 11-1 in step 1 was replaced with compound 15-1. The title compound 15 was prepared by the same method.

[0436] Example 16: 1-(2,2-difluoroethyl)-4-methyl-6-(2-(2-(trifluoromethyl)pyridin-4-yl)-2,7-diazaspiro[3.5]nonane-7-yl)-1,4-dihydro-5H-pyrazolo[3,4-b]pyrazin-5-one

[0437] Step 1: Synthesis of intermediate 16-2:

[0438] Under nitrogen protection, compound 16-1 (10.0 g) and urea peroxide (9.00 g) were added to 200 mL of acetonitrile solution, followed by the addition of trifluoroacetic anhydride (19.2 g). The reaction mixture was heated to room temperature and stirred for 3 hours until the reaction was complete. 50 mL of 0.5 M hydrochloric acid aqueous solution was added to the reaction mixture, and the mixture was extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was then slurried using ethyl acetate and petroleum ether (ethyl acetate: petroleum ether = 1:1) to obtain the title compound 16-2 (7.20 g).

[0439] 1 H NMR (400MHz, CDCl3) δ8.33(s,1H),8.11(s,1H),6.39-6.10(m,1H),4.85-4.77(m,2H).

[0440] MS m / z(ESI):=235.0[M+H] +

[0441] Step 2: Synthesis of intermediate 16-3:

[0442] Under nitrogen protection, compound 16-2 (7.20 g) was added to 140 mL of chloroform, followed by trifluoroacetic anhydride (32.2 g) and trimethylchlorosilane (16.7 g). After stirring at room temperature for 1 hour, triethylamine (15.5 g) was added to the reaction solution, and the reaction was completed after stirring at room temperature for 16 hours. 150 mL of water was added to the reaction solution, and after stirring for 15 minutes, a yellow solid precipitated. The solid was collected by filtration. The filtrate was extracted with dichloromethane:methanol = 10:1, 100 mL * 3. The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The yellow solid and the crude product were combined and slurried with ethyl acetate:petroleum ether = 1:1 to obtain the title compound 16-3 (6.00 g).

[0443] 1 H NMR (400MHz, DMSO-d6) δ12.86(s,1H),8.30(s,1H),6.55-6.26(m,1H),4.81-4.73(m,2H).

[0444] MS m / z(ESI):=235.1[M+H] +

[0445] Step 3: Synthesis of intermediate 16-4:

[0446] Under nitrogen protection, compound 16-3 (6.00 g) and potassium carbonate (7.10 g) were dissolved in N,N-dimethylformamide (100 mL). Iodomethane (4.40 g) was added dropwise at 0 °C. After the addition was complete, the temperature was raised to 50 °C and stirred for 2 hours until the reaction was complete. 100 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (100.0 mL * 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was subjected to silica gel column chromatography (ethyl acetate / petroleum ether = 0%–100%) to obtain the title compound 16-4 (4.50 g).

[0447] 1 H NMR (400MHz, DMSO-d6) δ8.01(s,1H),6.56-6.29(m,1H),4.84-4.76(m,2H),3.59(s,3H).

[0448] MS m / z(ESI):=249.1[M+H] +

[0449] Step 4: Synthesis of intermediate 16-5:

[0450] Under a nitrogen atmosphere, compound 16-4 (150 mg), tert-butyl 2,7-diazaspiro[3.5]nonane-2-carboxylic acid (196 mg), and potassium carbonate (166 mg) were dissolved in N,N-dimethylformamide (3 mL). After stirring at room temperature for 2 hours, the reaction was completed. 10 mL of water was added to the reaction solution, and ethyl acetate (20 mL * 3) was used for extraction. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by silica gel column chromatography (dichloromethane / methanol = 10 / 1) to obtain the title compound 16-5 (200 mg).

[0451] Step 5: Synthesis of intermediate 16-6:

[0452] Compound 16-5 (200 mg) was dissolved in 6 mL of dichloromethane, and trifluoroacetic acid (2.00 mL) was slowly added dropwise at 0 °C. The reaction solution was stirred at room temperature for 2 hours until the reaction was complete. The reaction solution was concentrated under reduced pressure to obtain the title compound 16-6 (150 mg).

[0453] Step 6: Synthesis of Compound 16

[0454] 4-Bromo-2-(trifluoromethyl)pyridine (102 mg), compound 16-6 (150 mg), and potassium carbonate (170 mg) were dissolved in N,N-dimethylformamide (2 mL). The reaction was carried out after stirring at 80 °C for 10 hours. The reaction solution was cooled to room temperature, 50 mL of water was added, and the mixture was extracted with ethyl acetate (20 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol: 50 / 1; Rf: 0.6) and high performance liquid chromatography (column: YMC TAR-C18, 30 * 150 mm, 5 μm; mobile phase: [water (containing 7 mmol / L ammonium bicarbonate) - acetonitrile]; acetonitrile gradient: 70-85%) to obtain title compound 16 (80.0 mg).

[0455] 1 H NMR (400MHz, DMSO-d6) δ8.23(d,J=5.7Hz,1H),7.68(s,1H),6.73(d,J=2.2Hz,1H),6.62-6. 18(m,2H),4.70-4.55(m,2H),3.87(brs,4H),3.80(s,4H),3.48(s,3H),1.95-1.84(m,4H).

[0456] MS m / z(ESI): 484.0 [M+H] +

[0457] Examples 17-19:

[0458] Referring to the synthesis method of Example 16, except that 2,7-diazaspiro[3.5]nonane-2-carboxylic acid tert-butyl ester in step 4 was replaced with the starting material in the table below, and the following compounds 17-19 were synthesized in the same manner.

[0459] Example 20: 1-(2,2-difluoroethyl)-4-methyl-6-(6-(((6-(trifluoromethyl)pyridin-2-yl)oxy)methyl)-2-azaspiro[3.3]hept-2-yl)-1,4-dihydro-5H-pyrazolo[4,3-b]pyridin-5-one

[0460] Step 1: Synthesis of intermediate 20-2:

[0461] Compound 20-1 (200 mg) was dissolved in 3 mL of acetonitrile. Potassium tert-butoxide (197 mg) was slowly added at 0 °C, and the mixture was stirred for 5 minutes. Then, 2-bromo-6-(trifluoromethyl)pyridine (218 mg) was added, and the mixture was heated to 60 °C and stirred for 2 hours until the reaction was complete. The reaction solution was cooled to room temperature, 50 mL of water was added, and the mixture was extracted with ethyl acetate (20 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol = 50 / 1) to obtain the title compound 20-2 (300 mg).

[0462] Step 2: Synthesis of intermediate 20-3:

[0463] Compound 20-2 (200 mg) was dissolved in 2 mL of dichloromethane, and trifluoroacetic acid (0.50 mL) was slowly added dropwise at 0 °C. The reaction was carried out after stirring at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure to obtain the title compound 20-3 (200 mg).

[0464] Step 3: Synthesis of Compound 20

[0465] Under nitrogen protection, compound 20-3 (150 mg), compound 6-5 (170 mg), tris(dibenzylacetone)palladium (35.6 mg), 4,5-bisdiphenylphosphine-9,9-dimethyloxanthracene (44.9 mg) and cesium carbonate (379 mg) were added to 1,4-dioxane (2 mL). The reaction solution was stirred at 90 °C for 10 hours until the reaction was complete. The reaction solution was cooled to room temperature, 50 mL of water was added, and the mixture was extracted with ethyl acetate (20 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol = 50 / 1) and high performance liquid chromatography (column: YMC TAR-C18, 30 * 150 mm, 5 μm; mobile phase: [water (containing 7 mmol / L ammonium bicarbonate) - acetonitrile]; acetonitrile gradient: 50-80%) to obtain the title compound 20 (125 mg).

[0466] 1 H NMR (400MHz, DMSO-d6) δ7.97(t,J=7.9,7.9Hz,1H),7.65(s,1H),7.47(d,J=7.3 Hz,1H),7.15(d,J=8.4Hz,1H),6.59(s,1H),6.34(tt,J=54.9,54.9,3.8,3.8Hz, 1H),4.73(td,J=15.1,15.0,3.8Hz,2H),4.27(d,J=6.7Hz,2H),4.01(s,2H),3.9 4(s,2H),3.46(s,3H),2.69-2.57(m,1H),2.41-2.26(m,2H),2.18-2.00(m,2H).

[0467] MS m / z(ESI): 484.1 [M+H] +

[0468] Example 21: 1-(2,2-difluoroethyl)-4-methyl-6-(6-(((6-(trifluoromethyl)pyridin-2-yl)oxy)methyl)-2-azaspiro[3.3]hept-2-yl)-1,4-dihydro-5H-pyrazolo[3,4-b]pyrazin-5-one

[0469] Compound 16-4 (100 mg), compound 20-3 (180 mg), and potassium carbonate (170 mg) were added to N,N-dimethylformamide (2 mL). The reaction was carried out after heating to 80 °C and stirring for 10 hours. The reaction solution was cooled to room temperature, 50 mL of water was added, and the mixture was extracted with ethyl acetate (20 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol: 50 / 1; Rf: 0.6) and high performance liquid chromatography (column: YMC TAR-C18, 30 * 150 mm, 5 μm; mobile phase: [water (containing 7 mmol / L ammonium bicarbonate) - acetonitrile]; acetonitrile gradient: 50-80%) to obtain title compound 21 (50.0 mg).

[0470] 1 H NMR (400MHz, DMSO-d6) δ7.97(t,J=7.9Hz,1H),7.58(s,1H),7.47(d,J=7.3Hz,1H),7.14(d,J=8.5Hz,1H),6.37(tt,J=55.1,4.0Hz,1H ),4.71-4.47(m,4H),4.27(d,J=6.7Hz,2H),4.18-4.00(m,2H),3.41(s,3H),2.69-2.56(m,1H),2.44-2.30(m,2H),2.19-1.97(m,2H).

[0471] MS m / z (ESI): 485.1 [M+H] +

[0472] Example 22: 1-(2,2-difluoroethyl)-4-methyl-6-(2-(2-(trifluoromethyl)pyrimidin-5-yl)-2,7-diazaspiro[3.5]non-7-yl)-1,4-dihydro-5H-pyrazolo[4,3-b]pyridin-5-one

[0473] Step 1: Synthesis of intermediate 22-2:

[0474] Under a nitrogen atmosphere, 1.39 g of 2-tert-butoxycarbonyl-2,7-diazaspiro[3.5]nonane, 1.50 g of 6-5, 5.02 g of cesium carbonate, 940 mg of tris(dibenzylacetone)dipalladium, and 594 mg of 4,5-bis(diphenylphosphine-9,9-dimethyloxanthracene) were added to 1,4-dioxane (15 mL), and the mixture was heated to 90 °C and stirred overnight. The reaction mixture was monitored by LCMS to ensure complete reaction. 50 mL of water was added to the reaction mixture, and the mixture was then extracted with ethyl acetate (50 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (dichloromethane / methanol: 50 / 1; Rf: 0.6) to obtain the title compound 22-2 (2.25 g).

[0475] Step 2: Synthesis of intermediate 22-3:

[0476] Compound 22-2 (0.88 g) was added to a TFA / DCM mixture of 1 / 4 (5.0 mL). The reaction was completed after stirring at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure to obtain the title compound 22-3 (0.50 g), which was used directly in the next reaction without further purification.

[0477] Step 3: Synthesis of Compound 22:

[0478] Intermediate 22-3 (87.0 mg), 5-bromo-2-(trifluoromethyl)pyrimidine (70.0 mg), and potassium carbonate (100 mg) from the previous step were added to N,N-dimethylformamide (2 mL). The reaction was carried out after heating to 80 °C and stirring for 2 hours. The reaction solution was cooled to room temperature, 50 mL of water was added, and the mixture was extracted with ethyl acetate (20 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol: 50 / 1; Rf: 0.6) and high performance liquid chromatography (column: YMC TAR-C18, 30 * 150 mm, 5 μm; mobile phase: [water (containing 7 mmol / L ammonium bicarbonate) - acetonitrile]; acetonitrile gradient: 70-85%) to obtain the title compound 22 (50.0 mg).

[0479] 1 H NMR(400MHz,DMSO-d6)δ8.16(s,2H),7.76(s,1H),7.28(s,1H),6.37(tt,J=54.9,3.6Hz,1H), 4.83(td,J=15.1,3.5Hz,2H),3.87(s,4H),3.51(s,3H),3.10(brs,4H),1.94(t,J=5.2Hz,4H).

[0480] MS m / z (ESI): 484.4 [M+H] +

[0481] Examples 23-29:

[0482] The following compounds 23-29 were synthesized using the same method as in Example 22, except that 5-bromo-2-(trifluoromethyl)pyrimidine in the reaction was replaced with the starting materials in the table below.

[0483] Example 30: 1-(2,2-difluoroethyl)-4-methyl-6-((2-(2-(trifluoromethyl)pyridin-4-yl)-2-azaspiro[3.5]nonane-7-yl)oxy)-1,4-dihydro-5H-pyrazolo[3,4-b]pyrazin-5-one

[0484] Step 1: Synthesis of intermediate 30-2:

[0485] Compound 30-1 (200 mg) was dissolved in 5 mL of tetrahydrofuran. Potassium tert-butoxide (200 mg) was added at 0 °C, and the mixture was stirred for 5 minutes. Then, compound 16-4 (200 mg) was added, and the mixture was heated to 60 °C and stirred for 2 hours until the reaction was complete. The reaction solution was cooled to room temperature, 50 mL of water was added, and the mixture was extracted with ethyl acetate (20 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol: 50 / 1) to obtain the title compound 30-2 (200 mg).

[0486] Step 2: Synthesis of intermediate 30-3:

[0487] Compound 30-2 (200 mg) was dissolved in 6 mL of dichloromethane, and trifluoroacetic acid (1.50 mL) was slowly added dropwise at 0 °C. The reaction solution was stirred at room temperature for 2 hours until the reaction was complete. The reaction solution was concentrated under reduced pressure to obtain the title compound 30-3 (150 mg).

[0488] Step 3: Synthesis of Compound 30:

[0489] 2-Trifluoromethyl-4-bromopyridine (102 mg), compound 30-3 (150 mg), and potassium carbonate (200 mg) were dissolved in N,N-dimethylformamide (2 mL). The reaction was carried out after heating to 80 °C and stirring for 10 hours. The reaction solution was cooled to room temperature, 50 mL of water was added, and the mixture was extracted with ethyl acetate (20 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by high performance liquid chromatography (column: YMC TAR-C18, 30 * 150 mm, 5 μm; mobile phase: [water (containing 7 mmol / L ammonium bicarbonate) - acetonitrile]; acetonitrile gradient: 60-90%) to obtain title compound 30 (40.0 mg).

[0490] 1 H NMR (400MHz, DMSO-d6) δ8.22(d,J=5.2Hz,1H),7.79(d,J=1.9Hz,1H),6.72(s,1H),6.59-6.20(m,2H),5.12-5 .05(m,1H),4.81-4.58(m,2H),3.78(s,2H),3.74(s,2H),3.51(s,3H),2.00-1.90(m,4H),1.83-1.58(m,4H).

[0491] MS m / z (ESI): 499.2 [M+H] +

[0492] Examples 31-34:

[0493] The synthesis method of Example 30 is referenced, except that 30-1 in step 1 is replaced with the starting materials in the table below, and the following compounds 31-34 are synthesized in the same way.

[0494] Example 35: 1-(2,2-difluoroethyl)-4-methyl-6-((7-(2-(trifluoromethyl)pyridin-4-yl)-7-azaspiro[3.5]nonane-2-yl)oxy)-1,4-dihydro-5H-pyrazolo[4,3-b]pyridin-5-one

[0495] Step 1: Synthesis of compound 35-2:

[0496] Under a nitrogen atmosphere, compound 34-1 (446 mg), compound 6-5 (270 mg), cesium carbonate (903 mg), and Rockphos Pd G3 (155 mg) were added to 1,4-dioxane (5 mL). The mixture was heated to 60 °C and stirred for 10 hours until the reaction was complete. 10 mL of water was added to the reaction solution, followed by extraction with ethyl acetate (30 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol: 10 / 1; Rf: 0.6) to obtain the title compound 35-2 (160 mg).

[0497] Step 2: Synthesis of compound 35-3:

[0498] Compound 35-2 (130 mg) was added to an ethyl acetate solution of hydrogen chloride (4 M, 10 mL) under a nitrogen atmosphere. The reaction was completed after stirring at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure, and the residue was slurried with ethyl acetate, filtered, and the filter cake was dried to obtain the title compound 35-3 (110 mg).

[0499] Step 3: Synthesis of Compound 35:

[0500] Compound 35-3 (100 mg) and 4-bromo-2-trifluoromethylpyridine (128 mg) were dissolved in N,N-dimethylformamide (2 mL), potassium carbonate (117 mg) was added, and the mixture was heated to 90 °C and stirred for 2 hours until the reaction was complete. The reaction solution was cooled to room temperature, 10 mL of water was added, and then extracted with ethyl acetate (20 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol: 10 / 1; Rf: 0.6) and high performance liquid chromatography (column: YMC TAR-C18, 30 * 150 mm, 5 μm; mobile phase: [water (containing 7 mmol / L ammonium bicarbonate) - acetonitrile]; acetonitrile gradient: 40-70%) to obtain the title compound 35 (36.0 mg).

[0501] 1 H NMR (400MHz, DMSO-d6) δ8.24(d,J=6.0Hz,1H),7.79(s,1H),7.34-7.12(m,2H),7.11-6.98(m,1H),6.37(tt,J=54.8,3.7Hz,1H), 5.08-4.52(m,3H),3.52(s,3H),3.52-3.44(m,2H),3.44-3.36(m,2H),2.61-2.52(m,2H),1.95-1.83(m,2H),1.76-1.57(m,4H).

[0502] MS m / z (ESI): 498.2 [M+H] + .

[0503] Examples 36-39

[0504] Referring to the synthesis method of Example 35, except that compound 34-1 in step 1 is replaced with the starting material in the table below, the following compounds 36-39 are synthesized in the same way.

[0505] Example 40: 1-(2,2-difluoroethyl)-4-methyl-6-(2-(((3-(trifluoromethyl)pyridin-4-yl)oxy)methyl)-7-azaspiro[3.5]non-7-yl)-1,4-dihydro-5H-pyrazolo[3,4-b]pyrazin-5-one

[0506] Step 1: Synthesis of intermediate 40-2:

[0507] Compound 40-1 (500 mg) was dissolved in 6 mL of acetonitrile. Potassium tert-butoxide (439 mg) was slowly added at 0 °C. After stirring for 5 minutes, 4-bromo-3-(trifluoromethyl)pyridine (486 mg) was added. The reaction mixture was stirred at 60 °C for 2 hours until the reaction was complete. After cooling to room temperature, 100 mL of water was added, and the mixture was extracted with ethyl acetate (30 mL x 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol: 50 / 1; Rf: 0.6) to obtain the title compound 40-2 (380 mg).

[0508] Step 2: Synthesis of intermediate 40-3:

[0509] Compound 40-2 (350 mg) was dissolved in 2 mL of dichloromethane, followed by the addition of a 1,4-dioxane solution of hydrogen chloride (4 M, 2 mL). The reaction was stirred at room temperature for 2 hours until complete. The reaction solution was concentrated under reduced pressure, slurried with ethyl acetate, and filtered. The filter cake was washed with ethyl acetate and dried to obtain the title compound 40-3 (280 mg).

[0510] Step 3: Synthesis of Compound 40:

[0511] Compound 40-3 (70.0 mg) was dissolved in N,N-dimethylformamide (1.00 mL), followed by the addition of potassium carbonate (86.2 mg). After stirring the reaction mixture for 5 minutes, compound 16-4 (51.7 mg) was added. The reaction was carried out after heating to 80 °C and stirring for 10 hours. The reaction mixture was cooled to room temperature, 50 mL of water was added, and the mixture was extracted with ethyl acetate (20 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol: 50 / 1; Rf: 0.6) and high performance liquid chromatography (column: YMC TAR-C18, 30 * 150 mm, 5 μm; mobile phase: [water (containing 7 mmol / L ammonium bicarbonate) - acetonitrile]; acetonitrile gradient: 65-95%) to obtain the title compound 40 (65.0 mg).

[0512] 1 H NMR (400MHz, DMSO-d6) δ8.70(t,J=3.0Hz,2H),7.64(s,1H),7.35(d,J=6.0Hz,1H),6.38(tt,J=55.0,3.9Hz,1H),4.60(td,J=14.8,3.9Hz ,2H),4.23(d,J=5.6Hz,2H),3.97-3.66(m,4H),3.46(s,3H),2.91-2.68(m,1H),2.08-1.86(m,2H),1.86-1.65(m,4H),1.66-1.52(m,2H).

[0513] MS m / z (ESI): 513.1 [M+H] +

[0514] Example 41: 1-(2,2-difluoroethyl)-4-methyl-6-(2-(((3-(trifluoromethyl)pyridin-4-yl)oxy)methyl)-7-azaspiro[3.5]non-7-yl)-1,4-dihydro-5H-pyrazolo[4,3-b]pyridin-5-one

[0515] The title compound 41 was prepared by referring to the synthesis method of Example 40, except that compound 16-4 in step 3 was replaced with compound 6-5.

[0516] 1H NMR (400MHz, DMSO-d6) δ8.70(t,J=2.9Hz,2H),7.74(s,1H),7.34(d,J=5.9Hz,1H),7.25(s,1H),6.36(tt,J=54.9,3.7Hz,1H),4.89-4.72(m,2H),4 .22(d,J=5.4Hz,2H),3.50(s,3H),3.10(brs,2H),2.98(brs,2H),2.85-2 .72(m,1H),2.01-1.84(m,2H),1.81-1.69(m,4H),1.65(t,J=5.5Hz,2H).

[0517] MS m / z (ESI): 512.1 [M+H] +

[0518] Example 42: 1-(2,2-difluoroethyl)-7-((3aR,5r,6aS)-2-(2-(trifluoromethyl)pyridin-4-yl)octahydrocyclopentan[c]pyrrolo-5-yl)-4,5,6,7-tetrahydropyrazolo[3,4-c]azapyro-8(1H)-one

[0519] Step 1: Synthesis of intermediate 42-2:

[0520] ((3aR,5r,6aS)-octahydrocyclopentan[c]pyrrolo-5-yl)tert-butyl carbamate (1.00 g) and 4-bromo-2-trifluoromethylpyridine (1.41 g) were dissolved in 20 mL of N,N-dimethylformamide, and cesium carbonate (4.07 g) was added. The mixture was heated to 90 °C and stirred for 6 hours until the reaction was complete. The reaction solution was cooled to room temperature, 100 mL of water was added, and the mixture was extracted with ethyl acetate (100 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (petroleum ether / ethyl acetate: 1 / 1) to obtain the title compound 42-2 (1.20 g).

[0521] Step 2: Synthesis of intermediate 42-3:

[0522] Under nitrogen protection, compound 42-2 (1.80 g) was added to a 1,4-dioxane solution (4 M, 30 mL) of hydrogen chloride and stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by slurrying with ethyl acetate to obtain the title compound 42-3 (1.00 g).

[0523] Step 3: Synthesis of intermediate 42-4:

[0524] 500 mg of methyl 4-bromo-1-(2,2-difluoroethyl)-1H-pyrazole-5-carboxylate was added to 5 mL of tetrahydrofuran, followed by compound 42-3 (504 mg) and 1,8-diazabicyclo[5.4.0]undec-7-ene (565 mg). The mixture was heated to 80 °C and stirred for 48 hours until the reaction was complete. 30 mL of water was added to the reaction mixture. The mixture was extracted with ethyl acetate (20 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (petroleum ether / ethyl acetate: 2 / 1) to obtain the title compound 42-4 (280 mg).

[0525] MS m / z (ESI): 508.3 [M+H] +

[0526] Step 4: Synthesis of intermediate 42-5:

[0527] Compound 42-4 (250 mg), (E)-3-(tert-butyldimethylsiloxy)propen-1-yl-boronic acid pinacol ester (176 mg), 1,1-bis(diphenylphosphine)ferrocene palladium dichloride (35.9 mg), and potassium carbonate (135 mg) were added to a mixed solution of 1,4-dioxane (5 mL) and water (1 mL). Under nitrogen protection, the mixture was heated to 100 °C and stirred for 2 hours until the reaction was complete. The reaction solution was cooled to room temperature, 10 mL of water was added, and the mixture was extracted with ethyl acetate (10 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (petroleum ether / ethyl acetate: 2 / 1) to obtain the title compound 42-5 (180 mg).

[0528] MS m / z (ESI): 600.4 [M+H] +

[0529] Step 5: Synthesis of intermediate 42-6:

[0530] Compound 42-5 (180 mg) was added to 5 mL of methanol, and palladium on carbon (10%, 10.8 mg) was added. The reaction was carried out under hydrogen gas and stirred at room temperature for 2 hours until completion. The reaction solution was filtered, the filter cake was washed with methanol, and the organic phase was concentrated to obtain crude compound 42-6 (135 mg), which was directly used in the next reaction step.

[0531] MS m / z (ESI): 602.4 [M+H] +

[0532] Step 6: Synthesis of intermediate 42-7:

[0533] Compound 42-6 (135 mg) was added to a 1,4-dioxane solution (4 M, 2 mL) of hydrogen chloride at 0 °C, and the reaction was completed after stirring at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure to obtain crude compound 42-7 (70.0 mg), which was directly used in the next reaction.

[0534] MS m / z (ESI): 488.3 [M+H] +

[0535] Step 7: Synthesis of intermediate 42-8:

[0536] Compound 42-7 (60.0 mg) was added to 3 mL of dichloromethane, followed by triethylamine (13.7 mg) and then methanesulfonyl chloride (15.5 mg). The reaction was carried out after stirring at room temperature for 10 hours. 5 mL of water was added to the reaction mixture. The mixture was extracted with dichloromethane (5 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (dichloromethane / methanol: 10 / 1) to obtain the title compound 42-8 (30.0 mg).

[0537] MS m / z (ESI): 566.4 [M+H] +

[0538] Step 8: Synthesis of Compound 42:

[0539] Compound 42-8 (30.0 mg) was added to 2 mL of anhydrous tetrahydrofuran. Sodium hydride (3.00 mg, 60% purity) was added at 0 °C, and the mixture was heated to 45 °C and stirred for 48 hours until the reaction was complete. 5 mL of water was added to the reaction mixture. The aqueous phase was extracted with ethyl acetate (5 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative high-performance liquid chromatography (HPLC) (column: Pursuit XRs 10C18 19*250 mm*10 μm; mobile phase: [water (0.1% formic acid)-acetonitrile]; acetonitrile gradient: 60%-66%) to obtain the title compound 42 (10.0 mg).

[0540] 1H NMR(400MHz,MeOD)δ8.21(d,J=6.9Hz,1H),7.43(s,1H),7.19(d,J=2.5Hz,1H) ,6.93(dd,J=6.9,2.6Hz,1H),6.17(tt,J=55.8,3.9Hz,1H),5.02-4.71(m,3H), 3.94-3.77(m,2H),3.70-3.57(m,2H),3.36-3.32(m,2H),3.06-2.92(m,2H),2 .75(t,J=7.5Hz,2H),2.37-2.17(m,2H),2.10-1.97(m,2H),1.80-1.59(m,2H).

[0541] MS m / z (ESI): 470.4 [M+H] +

[0542] Example 43: 1-(2,2-difluoroethyl)-4-methyl-6-(2-(2-(trifluoromethyl)pyridin-4-yl)-2-azaspiro[3.5]non-6-en-7-yl)-1,4-dihydro-5H-pyrazolo[4,3-b]pyridin-5-one

[0543] Step 1: Synthesis of compound 43-2:

[0544] Compound 43-1 (0.50 g) was dissolved in 10 mL of dichloromethane, and trifluoroacetic acid (476 mg) was added. The mixture was stirred at room temperature for 2 hours until the reaction was complete. The reaction solution was concentrated to obtain crude compound 43-2 (500 mg), which was used directly in the next reaction.

[0545] m / z:ES + [M+H] + =140.1

[0546] Step 2: Synthesis of compound 43-3:

[0547] The crude compound 43-2 (500 mg) obtained in the previous step was dissolved in N,N-dimethylformamide (10 mL), potassium carbonate (682 mg) and 2-trifluoromethyl-4-bromopyridine (480 mg) were added, and the mixture was heated to 100 °C and stirred for 2 hours until the reaction was complete. The reaction solution was diluted with 10 mL of ethyl acetate, washed with saturated brine and dried over anhydrous sodium sulfate, filtered, concentrated, and the concentrate was purified by column chromatography (ethyl acetate / petroleum ether: 0-30%) to obtain the title compound 43-3 (150 mg).

[0548] m / z:ES + [M+H] +=285.1

[0549] 1 H NMR (400MHz, Chloroform-d) δ8.50-8.14(m,1H),6.71-6.57(m,1H),6.51-6.34(m,1H),4.01-3.84(m,4H),2.55-2.37(m,4H),2.25-2.06(m,4H).

[0550] Step 3: Synthesis of compound 43-4:

[0551] Compound 43-3 (150 mg) was dissolved in 5 mL of tetrahydrofuran. After cooling to -70 °C, a tetrahydrofuran solution of potassium bis(trimethylsilyl)amino (1 M, 0.8 mL) was slowly added. The reaction mixture was stirred at -70 °C for 30 minutes, and then a tetrahydrofuran solution of N-phenylbis(trifluoromethanesulfonyl)imide (0.5 M, 1.37 mL) was added dropwise. The reaction mixture was brought back to room temperature and stirred for 1 hour. The reaction was monitored by LC-MS and found to be complete. The reaction was quenched with water, diluted with 10 mL of ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (ethyl acetate / petroleum ether: 0-20%) to obtain the title compound 43-4 (160 mg).

[0552] Step 4: Synthesis of Compounds 43-5:

[0553] Compound 43-4 (160 mg), pinacol diboronate (254 mg), 1,1-bis(diphenylphosphine)ferrocene palladium dichloride (28.0 mg), and potassium acetate (113 mg) were added to 1,4-dioxane (3 mL). The mixture was heated to 100 °C and stirred overnight under nitrogen protection until the reaction was complete. The reaction solution was cooled to room temperature and diluted with ethyl acetate. After filtration, the filtrate was evaporated to dryness to obtain the title compound 43-5 (150 mg), which was used directly in the next reaction.

[0554] Step 5: Synthesis of Compound 43

[0555] Compound 43-5 (40.0 mg), compound 6-5 (30.0 mg), potassium carbonate (20 mg), and 1,1-bis(diphenylphosphine)ferrocene palladium dichloride (7.50 mg) were added to 1,4-dioxane (1 mL) and water (0.25 mL). The reaction was carried out after stirring at 100 °C for 1 hour. The reaction solution was diluted with 10 mL of ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by preparative liquid chromatography (HPLC) using high performance liquid chromatography (column: Boston Prime C18 column: 5 μm silica, 30 mm diameter, 150 mm length; mobile phase: [water (containing 7 mmol / L ammonium bicarbonate) - acetonitrile]; acetonitrile gradient: 50%-80%) to obtain the title compound 43 (10.0 mg).

[0556] 1 H NMR (400MHz, DMSO-d6) δ8.23(d,J=5.7Hz,1H),7.87(s,1H),7.84(d,J=0.8Hz,1H),6.76(d,J=2.3Hz,1H),6.63-6.53(m ,1H),6.54-6.17(m,2H),4.99-4.79(m,2H),3.94-3.70(m,4H),3.51(s,3H),2.55-2.51(m,4H),1.93(t,J=6.2Hz,2H).

[0557] MS m / z (ESI): 480.1 [M+H] +

[0558] Example 44: 1-(2,2-difluoroethyl)-4-methyl-6-(2-(2-(trifluoromethyl)pyridin-4-yl)-2-azaspiro[3.5]non-7-yl)-1,4-dihydro-5H-pyrazolo[4,3-b]pyridin-5-one

[0559] Compound 43 (40.0 mg) and palladium / carbon (10%, 10.1 mg) were added to 4 mL of methanol solution, and the reaction was carried out at room temperature for 6 hours under a hydrogen atmosphere. The reaction solution was filtered, the filter cake was washed with methanol, and the filtrates were combined to obtain the crude product. The title compound 44 (8.00 mg) was purified by preparative liquid chromatography (column: Boston Prime C18 column: 5 μm silica, 30 mm diameter, 150 mm length; mobile phase: [water (containing 7 mmol / L ammonium bicarbonate)-acetonitrile]; acetonitrile gradient: 50%-80%).

[0560] 1H NMR(400MHz,DMSO-d6)δ8.24(d,J=5.9Hz,1H),7.83(s,1H),7.81(s,1H),6.8 0(d,J=2.2Hz,1H),6.65-6.56(m,1H),6.39(tt,J=54.7,3.6Hz,1H),4.88(td, J=15.2,3.6Hz,2H),3.86(s,2H),3.76(s,2H),3.52(s,3H),2.84-2.78(m,1H) ,2.11-1.93(m,2H),1.87-1.75(m,2H),1.73-1.60(m,2H),1.46-1.32(m,2H).

[0561] MS m / z (ESI): 482.2 [M+H] +

[0562] Compound 45: 1,4-Dimethyl-6-(2-(2-(trifluoromethyl)pyridin-4-yl)-2,7-diazaspiro[3.5]non-7-yl)-1,4-dihydro-5H-pyrazolo[4,3-b]pyridin-5-one

[0563] Following the same synthetic method as compound 6, compound 45 was prepared using the above-described technical route.

[0564] 1 H NMR (400MHz, DMSO-d6) δ8.23(d,J=5.6Hz,1H),7.61(s,1H),7.17(s,1H),6.74(d,J=2.2Hz,1H),6.56 (dd,J=5.6,2.2Hz,1H),3.93(s,3H),3.80(s,4H),3.49(s,3H),3.10(brs,4H),1.91(t,J=5.1Hz,4H).

[0565] Example 46: 1-(2,2-difluoroethyl)-4-methyl-6-((7-(2-(trifluoromethyl)pyridin-4-yl)-7-azaspiro[3.5]nonane-2-yl)methoxy)-1,4-dihydro-5H-pyrazolo[3,4-b]pyrazin-5-one

[0566] Step 1: Synthesis of compound 46-2:

[0567] Compound 46-1 (250 mg) was dissolved in 5 mL of dichloromethane, and trifluoroacetic acid (1.00 mL) was slowly added dropwise at 0 °C. The reaction solution was stirred at room temperature for 2 hours until the reaction was complete. The reaction solution was concentrated under reduced pressure to obtain crude compound 46-2 (250 mg), which was used directly in the next step.

[0568] Step 2: Synthesis of compound 46-3:

[0569] Compound 46-2 (250 mg) was dissolved in N,N-dimethylformamide (3 mL), potassium carbonate (384 mg) was added, and after stirring for 5 minutes, 2-trifluoromethyl-4-bromopyridine (230 mg) was added. The reaction was carried out after heating to 60 °C and stirring for 10 hours. The reaction solution was cooled to room temperature, 50 mL of water was added, and then extracted with ethyl acetate (20 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol: 50 / 1) to obtain the title compound 46-3 (270 mg).

[0570] Step 3: Synthesis of Compound 46:

[0571] Compound 46-3 (75.6 mg) was dissolved in 1 mL of tetrahydrofuran, and potassium tert-butoxide (28.2 mg) was added. After stirring for 5 minutes, compound 16-4 (50.0 mg) was added. The reaction mixture was stirred at room temperature for 2 hours until the reaction was complete. 30 mL of water was added to the reaction mixture, and then it was extracted with ethyl acetate (20 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol: 50 / 1; Rf: 0.6) and high performance liquid chromatography (column: YMC TAR-C18, 30 * 150 mm, 5 μm; mobile phase: [water (containing 7 mmol / L ammonium bicarbonate) - acetonitrile]; acetonitrile gradient: 50-80%) to obtain the title compound 46 (20.0 mg).

[0572] 1 H NMR (400MHz, DMSO-d6) δ8.22(d,J=6.0Hz,1H),7.79(s,1H),7.19(d,J=2.6Hz,1H),7.08-6.98(m,1H),6.42(tt,J=54.8,3.8Hz,1H),4.68(td,J=14 .9,3.8Hz,2H),4.37(d,J=6.5Hz,2H),3.52(s,3H),3.47-3.39(m,2H),3. 39-3.33(m,2H),2.89-2.72(m,1H),2.10-1.91(m,2H),1.86-1.51(m,6H).

[0573] MS m / z (ESI): 513.2 [M+H] +

[0574] Example 47: 1-(2,2-difluoroethyl)-4-methyl-6-((7-(2-(trifluoromethyl)pyridin-4-yl)-7-azaspiro[3.5]nonane-2-yl)methoxy)-1,4-dihydro-5H-pyrazolo[4,3-b]pyridin-5-one

[0575] Compound 6-5 (194 mg), compound 46-3 (100 mg), methanesulfonate (2-(di-tert-butylphosphine)-3-methoxy-6-methyl,2,4,6-triisopropyl-1,1-biphenyl)(2-amino-1,1-biphenyl-2-yl)palladium(II) (55.9 mg), and cesium carbonate (325 mg) were added to 1,4-dioxane (2 mL). Under nitrogen protection, the mixture was heated to 100 °C and stirred for 10 hours until the reaction was complete. The reaction solution was cooled to room temperature, 30 mL of water was added, and then extracted with ethyl acetate (10 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol: 50 / 1; Rf: 0.6) and high performance liquid chromatography (column: YMC TAR-C18, 30 * 150 mm, 5 μm; mobile phase: [water (containing 7 mmol / L ammonium bicarbonate) - acetonitrile]; acetonitrile gradient: 55-85%) to obtain the title compound 47 (30.0 mg).

[0576] 1 H NMR (400MHz, DMSO-d6) δ8.24(d,J=6.0Hz,1H),7.78(d,J=0.8Hz,1H),7.45(s,1H),7.20(d,J=2.6Hz,1H),7.11-6.92(m,1H),6.66-6.18(m,1H),4.9 3-4.69(m,2H),3.98(d,J=6.4Hz,2H),3.52(s,3H),3.46-3.43(m,2H),3. 38-3.35(m,2H),2.87-2.73(m,1H),2.12-1.89(m,2H),1.82-1.55(m,6H).

[0577] MS m / z (ESI): 512.1 [M+H] +

[0578] Examples 48-51:

[0579] Referring to the synthesis method of Example 22, except that 5-bromo-2-(trifluoromethyl)pyrimidine in the reaction was replaced with the starting material in the table below, the following compounds 48-51 were synthesized in the same manner.

[0580] Examples 52-56:

[0581] Referring to the synthesis method of Example 42, except that 42-1 in step 1 is replaced with the starting materials in the table below, the following compounds 52-56 are synthesized in the same way.

[0582] Example 57: 4-Methyl-1-(oxecyclobutane-3-yl)-6-(2-(2-(trifluoromethyl)pyridin-4-yl)-2,7-diazaspiro[3.5]non-7-yl)-1,4-dihydro-5H-pyrazolo[4,3-b]pyridin-5-one

[0583] Step 1: Synthesis of intermediate 57-2:

[0584] Compound 57-1 (5.30 g) was added to 60 mL of N,N-dimethylformamide, along with 7.39 g of 3-iodooxetine and 26.2 g of potassium carbonate. The mixture was heated to 80 °C and stirred for 10 hours until the reaction was complete. The reaction solution was filtered, and the filtrate was concentrated and then subjected to silica gel column chromatography (petroleum ether: ethyl acetate = 2:1) to obtain the title compound 57-2 (2.20 g).

[0585] MS m / z(ESI): 254.0 [M+H] +

[0586] Step 2: Synthesis of intermediate 57-3:

[0587] Compound 57-2 (2.20 g) was dissolved in 30 mL of acetonitrile solution, and urea peroxide (2.04 g) and trifluoroacetic anhydride (3.64 g) were added. The reaction was stirred at room temperature for 4 hours until the reaction was complete. The reaction solution was filtered, the filter cake was washed with acetonitrile, and purified by silica gel column chromatography (dichloromethane:methanol = 10:1) to obtain the title compound 57-3 (1.80 g).

[0588] Step 3: Synthesis of intermediate 57-4:

[0589] Compound 57-3 (1.80 g) was added to 20 mL of acetic anhydride, heated to 140 °C and stirred under reflux for 2 hours. The temperature was then lowered to 90 °C, and 20 mL of water was added. The reaction was allowed to proceed for 2 hours, and LC-MS showed completion. The reaction solution was cooled to room temperature, and the pH was adjusted to 7–8 with 2 M sodium hydroxide solution. Extraction was performed with ethyl acetate (30 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane:methanol = 10:1) to obtain the title compound 57-4 (600 mg).

[0590] MS m / z(ESI): 270.1 [M+H] +

[0591] Step 4: Synthesis of intermediate 57-5:

[0592] Compound 57-4 (600 mg) was added to N,N-dimethylformamide (6 mL), along with potassium carbonate (919 mg) and methyl iodide (73.0 mg). The mixture was stirred at room temperature for 1 hour until the reaction was complete. The reaction solution was added to 20 mL of water and extracted with ethyl acetate (20 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane:methanol = 10:1) to obtain the title compound 57-5 (120 mg).

[0593] MS m / z(ESI): 284.1 [M+H] +

[0594] Step 5: Synthesis of intermediate 57-6:

[0595] Compound 57-5 (70.0 mg) was added to a 1,4-dioxane (2 mL) solution, followed by Xphos Pd G4 (24.0 mg), potassium carbonate (241 mg), and tert-butyl 2,7-diazaspiro[3.5]nonane-2-carboxylic acid (80.0 mg). The mixture was heated to 100 °C and stirred for 2 hours until the reaction was complete. The reaction solution was concentrated and 20 mL of water was added. The mixture was extracted with ethyl acetate (20 mL * 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (after adding the reaction solution and filtering, the residue was purified by column chromatography (dichloromethane:methanol = 10:1) to obtain the title compound 57-6 (50.0 mg).

[0596] Step 6: Synthesis of intermediate 57-7:

[0597] Compound 57-6 (50.0 mg) was added to a 4M ethyl acetate solution (2 mL), and the reaction was carried out by stirring at 25 °C for 1 hour. The reaction solution was then concentrated under reduced pressure to obtain crude compound 57-7 (30.0 mg).

[0598] Step 7: Synthesis of Compound 57

[0599] Compound 57-7 (30.0 mg) and 4-bromo-2-(trifluoromethyl)pyridine (30.2 mg) were dissolved in N,N-dimethylformamide (3 mL), and potassium carbonate (50.0 mg) was added. The reaction mixture was stirred at 80 °C for 2 hours until the reaction was complete. The reaction mixture was cooled to room temperature, 10 mL of water was added, and the mixture was extracted with ethyl acetate (20 mL * 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol: 10 / 1) and preparative high performance liquid chromatography (column: Phenomenex Gemini NX 150 × 30 mm, 5 μm; mobile phase: [water (containing 7 mmol / L ammonium bicarbonate) - acetonitrile]; acetonitrile gradient: 50%-90%) to obtain the title compound 57 (7.08 mg).

[0600] 1 H NMR (400MHz, DMSO-d6) δ8.23(d,J=5.7Hz,1H),7.83(s,1H),7.22(s,1H),6.74(d,J=2.1Hz,1H),6.60-6.52(m,1H ),6.05-5.86(m,1H),4.95(s,2H),4.94(s,2H),3.79(s,4H),3.52(s,3H),3.10(brs,4H),1.91(t,J=5.4Hz,4H).

[0601] MS m / z (ESI): 475.3 [M+H] +

[0602] Examples 58-59:

[0603] The synthesis method of Example 57 was followed, except that 3-iodooxetine in step 1 was replaced with the starting material in the table below, and compounds 58-59 were prepared by the same method.

[0604] Example 60: 1-(2,2-difluoroethyl)-4-methyl-6-(2-(2-(trifluoromethyl)pyridin-4-yl)-2,8-diazaspiro[4.5]decane-8-yl)-1,4-dihydro-5H-pyrazolo[4,3-b]pyridin-5-one

[0605] Step 1: Synthesis of intermediate 60-2:

[0606] Under a nitrogen atmosphere, compound 6-5 (200 mg), cesium carbonate (600 mg), tert-butyl 2,8-diazaspiro[4.5]decane-2-carboxylate (200 mg), tris(dibenzylacetone)dipalladium (60.0 mg), and 4,5-bis(diphenylphosphine-9,9-dimethyloxanthracene) (70.0 mg) were added to 5 mL of 1,4-dioxane. The mixture was heated to 90 °C and stirred for 2 hours until the reaction was complete. The reaction solution was cooled to room temperature, 20 mL of water was added, and the mixture was extracted with ethyl acetate (30.00 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol: 10 / 1) to obtain the title compound 60-2 (100 mg). Step 2: Synthesis of intermediate 60-3:

[0607] Compound 60-2 (100 mg) was added to a 4 M ethyl acetate solution of hydrogen chloride (10 mL), and the reaction was carried out by stirring at 25 °C for 2 hours. The reaction solution was then concentrated under reduced pressure to obtain crude compound 60-3 (50.0 mg).

[0608] Step 3: Synthesis of Compound 60:

[0609] Compound 60-3 (50.0 mg) and 4-bromo-2-(trifluoromethyl)pyridine (40.5 mg) were dissolved in N,N-dimethylformamide (3 mL), and potassium carbonate (60.0 mg) was added. The reaction mixture was stirred at 80 °C for 2 hours until the reaction was complete. The reaction mixture was cooled to room temperature, 10 mL of water was added, and the mixture was extracted with ethyl acetate (20 mL * 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol: 10 / 1) and preparative high performance liquid chromatography (column: Phenomenex Gemini NX 150 × 30 mm, 5 μm; mobile phase: [water (containing 7 mmol / L ammonium bicarbonate) - acetonitrile]; acetonitrile gradient: 50%-90%) to obtain the title compound 60 (20.0 mg).

[0610] 1H NMR (400MHz, DMSO) δ8.21(d,J=5.8Hz,1H),7.76(s,1H),7.27(s,1H),6.87(s,1H),6.69(dd,J=5.9,2.3Hz,1H),6.59-6.15(m,1H),4.82(t d,J=15.1,3.7Hz,2H),3.51(s,3H),3.45(t,J=7.0Hz,2H),3.39-3.16(m,4H),3.15-3.06(m,2H),1.94(t,J=7.0Hz,2H),1.80-1.61(m,4H).

[0611] MS m / z (ESI): 497.5 [M+H] +

[0612] Example 61: 1-(2,2-difluoroethyl)-4-methyl-6-((7-(2-(trifluoromethyl)pyrimidin-5-yl)-7-azaspiro[3.5]non-2-yl)oxy)-1,4-dihydro-5H-pyrazolo[3,4-b]pyrazin-5-one

[0613] Step 1: Synthesis of intermediate 61-2:

[0614] Compound 61-1 (250 mg) was dissolved in 2 mL of dichloromethane, and a 1,4-dioxane solution of hydrogen chloride (4 M, 2.00 mL) was slowly added dropwise at 0 °C. The reaction mixture was stirred at room temperature for 2 hours until the reaction was complete. The reaction mixture was concentrated under reduced pressure to obtain crude compound 61-2 (150 mg), which was used directly in the next step.

[0615] Step 2: Synthesis of intermediate 61-3:

[0616] Compound 61-2 (150 mg) was dissolved in N,N-dimethylformamide (3 mL), potassium carbonate (300 mg) was added, and the mixture was stirred for 5 minutes. Then, 5-bromo-2-trifluoromethylpyrimidine (220 mg) was added. The reaction was carried out after heating to 60 °C and stirring for 10 hours. The reaction solution was cooled to room temperature, 50 mL of water was added, and the mixture was extracted with ethyl acetate (20 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol: 50 / 1) to obtain the title compound 61-3 (200 mg).

[0617] Step 3: Synthesis of Compound 61:

[0618] Compound 61-3 (100 mg) was dissolved in 2 mL of tetrahydrofuran, and potassium tert-butoxide (50.0 mg) was added. After stirring for 5 minutes, compound 16-4 (50.0 mg) was added. The reaction mixture was stirred at room temperature for 2 hours until the reaction was complete. 30 mL of water was added to the reaction mixture, and then it was extracted with ethyl acetate (20 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol: 50 / 1; Rf: 0.6) and high performance liquid chromatography (column: YMC TAR-C18, 30 * 150 mm, 5 μm; mobile phase: [water (containing 7 mmol / L ammonium bicarbonate) - acetonitrile]; acetonitrile gradient: 50-80%) to obtain the title compound 61 (10.0 mg).

[0619] 1 H NMR (400MHz, DMSO-d6) δ8.61 (s, 2H), 7.79 (s, 1H), 6.66-6.17 (m, 1H), 5.35-5.08 (m, 1H), 4.68 (td, J = 15. 1,3.4Hz,2H),3.52(s,3H),3.50-3.36(m,4H),2.56-2.52(m,2H),2.07-1.93(m,2H),1.85-1.66(m,4H).

[0620] MS m / z (ESI): 500.4 [M+H] +

[0621] Example 62: 1-(2,2-difluoroethyl)-4-methyl-6-((2-(2-(trifluoromethyl)pyrimidin-5-yl)-2-azaspiro[3.4]oct-6-yl)oxy)-1,4-dihydro-5H-pyrazolo[3,4-b]pyrazin-5-one

[0622] The title compound 62 was prepared by referring to the synthesis of Example 61, except that 61-1 in step 1 was replaced with 62-1.

[0623] 1 H NMR(400MHz, DMSO-d6)δ8.12(s,2H),7.79(s,1H),6.41(tt,J=54.8,3.7Hz,1H),5.51-5.30(m,1H),4.69(td, J=15.0,3.7Hz,2H),4.18-3.94(m,4H),3.51(s,3H),2.48-2.36(m,1H),2.29-2.06(m,3H),2.04-1.79(m,2H).

[0624] MS m / z (ESI): 486.3 [M+H] +

[0625] Example 63: 1-(2,2-difluoroethyl)-4-methyl-6-((2-(2-(trifluoromethyl)pyrimidin-5-yl)-2-azaspiro[3.5]non-7-yl)oxy)-1,4-dihydro-5H-pyrazolo[3,4-b]pyrazin-5-one

[0626] The title compound 63 was prepared by referring to the synthesis of Example 61, except that 61-1 in step 1 was replaced with 63-1.

[0627] 1 H NMR(400MHz,DMSO-d6)δ8.14(s,2H),7.79(s,1H),6.60-6.23(m,1H),5.13-5.05(m,1H),4.8 0-4.54(m,2H),3.85(s,2H),3.81(s,2H),3.51(s,3H),2.00-1.97(m,4H),1.87-1.53(m,4H).

[0628] MS m / z (ESI): 500.4 [M+H] +

[0629] Examples 64-66:

[0630] The synthesis method of Example 46 is referenced, except that 46-1 in step 1 is replaced with the starting material in the table below, and compounds 64-66 are prepared by the same method.

[0631] Example 67: 1-(2,2-difluoroethyl)-4-methyl-6-(2-(2-(trifluoromethyl)pyridin-4-yl)-2-azaspiro[3.5]non-6-en-7-yl)-1,4-dihydro-5H-pyrazolo[3,4-b]pyrazin-5-one

[0632] Compound 43-5 (40.0 mg), compound 16-4 (30.0 mg), potassium carbonate (20 mg), and 1,1-bis(diphenylphosphine)ferrocene palladium dichloride (7.50 mg) were added to 1,4-dioxane (1 mL) and water (0.25 mL). The reaction was carried out after stirring at 100 °C for 1 hour. The reaction solution was diluted with 10 mL of ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by preparative liquid chromatography (HPLC) using high performance liquid chromatography (column: Boston Prime C18 column: 5 μm silica, 30 mm diameter, 150 mm length; mobile phase: [water (containing 7 mmol / L ammonium bicarbonate) - acetonitrile]; acetonitrile gradient: 50%-80%) to obtain the title compound 67 (20.0 mg).

[0633] 1 H NMR (400MHz, DMSO-d6) δ8.22(d,J=5.6Hz,1H),7.92(s,1H),7.86-7.73(m,1H),6.73(d,J=2.2Hz,1H),6.66 -6.24(m,2H),4.97-4.70(m,2H),3.96-3.69(m,4H),3.55(s,3H),2.73-2.55(m,4H),1.96(t,J=6.1Hz,2H).

[0634] MS m / z (ESI): 481.1 [M+H] +

[0635] Example 68: 1-(2,2-difluoroethyl)-4-methyl-6-(2-(2-(trifluoromethyl)pyridin-4-yl)-2-azaspiro[3.5]non-7-yl)-1,4-dihydro-5H-pyrazolo[3,4-b]pyrazin-5-one

[0636] Compound 67 (30.0 mg) and palladium / carbon (10%, 10.1 mg) were added to 4 mL of methanol solution, and the reaction was carried out at room temperature for 6 hours under a hydrogen atmosphere. The reaction solution was filtered, the filter cake was washed with methanol, and the filtrates were combined to obtain the crude product. The title compound 68 (10.0 mg) was purified by preparative liquid chromatography (column: Boston Prime C18 column: 5 μm silica, 30 mm diameter, 150 mm length; mobile phase: [water (containing 7 mmol / L ammonium bicarbonate)-acetonitrile]; acetonitrile gradient: 50%-80%).

[0637] 1H NMR (400MHz, DMSO-d6) δ8.21(d,J=5.7Hz,1H),7.90(s,1H),6.74(d,J=2.2Hz,1H),6.65-6.10(m,2H),4.96-4 .69(m,2H),3.79(s,2H),3.70(s,2H),3.54(s,3H),3.19-3.13(m,1H),2.11-1.76(m,4H),1.80-1.41(m,4H).

[0638] MS m / z (ESI): 483.1 [M+H] +

[0639] Example 69: 1-(2,2-difluoroethyl)-4-methyl-6-(2-(2-(trifluoromethyl)pyrimidin-5-yl)-2,6-diazaspiro[3.4]oct-6-yl)-1,4-dihydro-5H-pyrazolo[3,4-b]pyrazin-5-one

[0640] Step 1: Synthesis of intermediate 69-2:

[0641] 1.00 g of tert-butyl 2,6-diazaspiro[3,4]octane-6-carboxylic acid and 1.40 g of 5-bromo-2-(trifluoromethyl)pyrimidine were dissolved in 20 mL of N,N-dimethylformamide, and 4.00 g of cesium carbonate was added. The mixture was heated to 90 °C and stirred for 6 hours until the reaction was complete. The reaction solution was cooled to room temperature, 100 mL of water was added, and the mixture was extracted with ethyl acetate (100 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (petroleum ether / ethyl acetate: 1 / 1) to obtain the title compound 69-2 (1.00 g).

[0642] Step 2: Synthesis of intermediate 69-3:

[0643] Under nitrogen protection, intermediate 69-2 (1.80 g) was added to an ethyl acetate solution of hydrogen chloride (4 M, 30 mL). The reaction mixture was stirred at room temperature for 2 hours until the reaction was complete. The reaction mixture was concentrated under reduced pressure, and the residue was purified by slurrying with ethyl acetate to obtain the title compound 69-3 (1.00 g).

[0644] Step 3: Synthesis of Compound 69:

[0645] Compound 16-4 (100 mg), compound 69-3 (180 mg), and potassium carbonate (170 mg) were added to N,N-dimethylformamide (2 mL). The reaction was carried out after stirring at 80 °C for 10 hours. The reaction solution was cooled to room temperature, 50 mL of water was added, and the mixture was extracted with ethyl acetate (20 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol: 50 / 1; Rf: 0.6) and high performance liquid chromatography (column: YMC TAR-C18, 30 * 150 mm, 5 μm; mobile phase: [water (containing 7 mmol / L ammonium bicarbonate) - acetonitrile]; acetonitrile gradient: 50-80%) to obtain the title compound 69 (60.0 mg).

[0646] 1 H NMR (400MHz, DMSO-d6) δ8.15(s,2H),7.57(s,1H),6.67-6.06(m,1H),4.79-4.44(m,2H),4.19-3.88(m,8H),3.43(s,3H),2.19(t,J=6.4Hz,2H).

[0647] MS m / z (ESI): 471.4 [M+H] +

[0648] Example 70: 1-(2,2-difluoroethyl)-4-methyl-6-(2-(2-(trifluoromethyl)pyrimidin-5-yl)-2,6-diazaspiro[3.4]oct-6-yl)-1,4-dihydro-5H-pyrazolo[4,3-b]pyridin-5-one

[0649] Under nitrogen protection, compound 69-3 (100 mg), compound 6-5 (150 mg), tris(dibenzylacetone)dipalladium (35.0 mg), 4,5-bisdiphenylphosphine-9,9-dimethyloxanthracene (45.0 mg) and cesium carbonate (300 mg) were added to 1,4-dioxane (2 mL). The reaction solution was stirred at 90 °C for 10 hours until the reaction was complete. The reaction solution was cooled to room temperature, 50 mL of water was added, and the mixture was extracted with ethyl acetate (20 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol = 50 / 1) and high performance liquid chromatography (column: YMC TAR-C18, 30 * 150 mm, 5 μm; mobile phase: [water (containing 7 mmol / L ammonium bicarbonate) - acetonitrile]; acetonitrile gradient: 50-80%) to obtain the title compound 70 (75.0 mg).

[0650] 1H NMR(400MHz,DMSO-d6)δ8.18(s,2H),7.67(s,1H),6.76(s,1H),6.61-5.89(m,1H),4.89-4.48 (m,2H),4.05(s,4H),3.78(s,2H),3.52(t,J=6.9Hz,2H),3.48(s,3H),2.21(t,J=6.9Hz,2H).

[0651] MS m / z (ESI): 470.3 [M+H] +

[0652] Example 71: 1-(2,2-difluoroethyl)-4-methyl-6-((2-(2-(trifluoromethyl)pyrimidin-5-yl)-2-azaspiro[3.4]oct-6-yl)oxy)-1,4-dihydro-5H-pyrazolo[4,3-b]pyridin-5-one

[0653] Step 1: Synthesis of intermediate 71-2:

[0654] 2-azaspiro[3.4]oct-6-ol (1.50 g) and 5-bromo-2-(trifluoromethyl)pyrimidine (1.40 g) were dissolved in 20 mL of N,N-dimethylformamide, and cesium carbonate (4.00 g) was added. The mixture was heated to 90 °C and stirred for 6 hours until the reaction was complete. The reaction solution was cooled to room temperature, 100 mL of water was added, and the mixture was extracted with ethyl acetate (100 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (petroleum ether / ethyl acetate: 1 / 1) to obtain the title compound 71-2 (1.20 g).

[0655] Step 2: Synthesis of Compound 71:

[0656] Under a nitrogen atmosphere, compound 71-2 (250 mg), compound 6-5 (270 mg), cesium carbonate (903 mg), and Rockphos Pd G3 (155 mg) were added to 1,4-dioxane (5 mL), and the mixture was heated to 80 °C and stirred for 10 hours until the reaction was complete. 10 mL of water was added to the reaction mixture, followed by extraction with ethyl acetate (30 mL x 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol: 10 / 1; Rf: 0.6) and high-performance liquid chromatography (column: YMC TAR-C18, 30 x 150 mm, 5 μm; mobile phase: [water (containing 7 mmol / L ammonium bicarbonate) - acetonitrile]; acetonitrile gradient: 50-80%) to obtain the title compound 71 (1.60 mg).

[0657] 1 H NMR(400MHz,DMSO-d6)δ8.13(s,2H),7.78(s,1H),7.38(s,1H),6.65-6.17(m ,1H),5.00-4.71(m,3H),4.18-3.91(m,4H),3.52(s,3H),2.45-1.73(m,6H).

[0658] MS m / z (ESI): 485.1 [M+H] +

[0659] Example 72: 1-(2,2-difluoroethyl)-4-methyl-6-(2-(2-(trifluoromethyl)pyrimidin-5-yl)-2-azaspiro[3.5]non-7-yl)-1,4-dihydro-5H-pyrazolo[4,3-d]pyrimidin-5,7(6H)-dione

[0660] The synthesis method of Example 1 was followed, except that 1-1 in step 1 was replaced with 72-1, and 4-bromo-2-trifluoromethylpyridine in step 6 was replaced with 5-bromo-2-trifluoromethylpyrimidine. Title compound 72 was prepared by the same method.

[0661] 1 H NMR(400MHz,DMSO-d6)δ8.17(s,2H),7.89(s,1H),6.61-6.14(m,1H),5.09-4.88(m,2H),4.79-4.77(m ,1H),3.88(s,2H),3.75(s,2H),3.39(s,3H),2.50-2.44(m,2H),2.07-2.94(m,2H),1.78-1.47(m,4H).

[0662] MS m / z (ESI): 500.4 [M+H] +

[0663] Example 73: 1-(2,2-difluoroethyl)-4-methyl-6-((2-(2-(trifluoromethyl)pyridin-4-yl)-2-azaspiro[3.5]non-7-yl)amino)-1,4-dihydro-5H-pyrazolo[3,4-b]pyrazin-5-one

[0664] Step 1: Synthesis of intermediate 73-2:

[0665] Under a nitrogen atmosphere, compound 73-1 (200 mg), compound 6-5 (250 mg), cesium carbonate (800 mg), and Xphos Pd G4 (120 mg) were added to 1,4-dioxane (5 mL). The mixture was heated to 80 °C and stirred for 10 hours until the reaction was complete. 10 mL of water was added to the reaction solution, followed by extraction with ethyl acetate (30 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol: 10 / 1) to obtain the title compound 73-2 (250 mg).

[0666] Step 2: Synthesis of intermediate 73-3:

[0667] Intermediate 73-2 (180 mg) was added to an ethyl acetate solution of hydrogen chloride (4 M, 5 mL), and the reaction was stirred at room temperature for 2 hours until the reaction was complete. The reaction solution was concentrated under reduced pressure to obtain crude compound 73-3 (150 mg).

[0668] Step 3: Synthesis of Compound 73:

[0669] Compound 73-3 (50.0 mg) and 4-bromo-2-(trifluoromethyl)pyridine (40.5 mg) were dissolved in N,N-dimethylformamide (3 mL), and potassium carbonate (60.0 mg) was added. The reaction mixture was stirred at 80 °C for 2 hours until the reaction was complete. The reaction mixture was cooled to room temperature, 10 mL of water was added, and the mixture was extracted with ethyl acetate (20 mL * 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol: 10 / 1) and preparative high performance liquid chromatography (column: Phenomenex Gemini NX 150 × 30 mm, 5 μm; mobile phase: [water (containing 7 mmol / L ammonium bicarbonate) - acetonitrile]; acetonitrile gradient: 50%-90%) to obtain the title compound 73 (20.0 mg).

[0670] 1 H NMR (400MHz, MeOD) δ8.14(d,J=5.8Hz,1H),7.67(s,1H),6.72(d,J=2.1Hz,1H),6.67(s,1H),6.57-6.47(m,1H),6.35-5.97(m,1H),4. 75-4.63(m,2H),3.82(s,2H),3.77(s,2H),3.67(s,3H),3.50-3.37(m,1H),2.15-1.97(m,4H),1.88-1.70(m,2H),1.57-1.38(m,2H).

[0671] MS m / z (ESI): 497.5 [M+H] +

[0672] Examples 74-75:

[0673] The synthesis method of compound 73 is referenced, except that 73-1 in step 1 is replaced with the starting material in the table below, and compounds 74-75 are prepared by the same method.

[0674] Example 76: 1-(2,2-difluoroethyl)-6-((2-(2-(difluoromethyl)pyridin-4-yl)-2-azaspiro[3.4]oct-6-yl)oxy)-4-methyl-1,4-dihydro-5H-pyrazolo[3,4-b]pyrazin-5-one

[0675] Step 1: Synthesis of intermediate 76-3:

[0676] Compound 71-1 (150 mg) was dissolved in N,N-dimethylformamide (3 mL), potassium carbonate (300 mg) was added, and after stirring for 5 minutes, 4-bromo-2-(difluoromethyl)pyridine (220 mg) was added. The reaction was carried out after heating to 60 °C and stirring for 10 hours. The reaction solution was cooled to room temperature, 50 mL of water was added, and then extracted with ethyl acetate (20 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol: 50 / 1) to obtain the title compound 76-3 (200 mg).

[0677] Step 3: Synthesis of Compound 76:

[0678] Compound 76-3 (100 mg) was dissolved in 2 mL of tetrahydrofuran, and potassium tert-butoxide (50.0 mg) was added. After stirring for 5 minutes, compound 16-4 (50.0 mg) was added. The reaction mixture was stirred at room temperature for 2 hours until the reaction was complete. 30 mL of water was added to the reaction mixture, and then it was extracted with ethyl acetate (20 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol: 50 / 1; Rf: 0.6) and high performance liquid chromatography (column: YMC TAR-C18, 30 * 150 mm, 5 μm; mobile phase: [water (containing 7 mmol / L ammonium bicarbonate) - acetonitrile]; acetonitrile gradient: 50-80%) to obtain the title compound 76 (40.0 mg).

[0679] 1H NMR (400MHz, DMSO-d6) δ8.17(d,J=5.6Hz,1H),7.79(s,1H),6.73(t,J=55.3Hz,1H),6.57-6.21(m,3H),5.51-5.32( m,1H),4.73-4.63(m,2H),4.06-3.80(m,4H),3.51(s,3H),2.46-2.36(m,1H),2.27-2.04(m,3H),2.02-1.80(m,2H).

[0680] MS m / z (ESI): 467.2 [M+H] +

[0681] Example 77: 1-(2,2-difluoroethyl)-6-(2-(2-(difluoromethyl)pyridin-4-yl)-2,6-diazaspiro[3.4]oct-6-yl)-4-methyl-1,4-dihydro-5H-pyrazolo[3,4-b]pyrazin-5-one

[0682] The title compound 77 was prepared by referring to the synthesis method of Example 69, except that 5-bromo-2-(trifluoromethyl)pyrimidine in step 1 was replaced with 4-bromo-2-(difluoromethyl)pyridine.

[0683] 1 H NMR (400MHz, DMSO-d6) δ8.19 (d, J = 5.6Hz, 1H), 7.59 (s, 1H), 7.03-6.15 (m, 4H), 4. 56(td,J=14.7,4.0Hz,2H),4.30-3.66(m,8H),3.44(s,3H),2.18(t,J=6.4Hz,2H).

[0684] MS m / z (ESI): 452.1 [M+H] +

[0685] Example 78: 1-(2,2-difluoroethyl)-4-methyl-6-(2-(2-(trifluoromethyl)pyrimidin-5-yl)-2,7-diazaspiro[3.5]non-7-yl)-1,4-dihydro-5H-pyrazolo[3,4-b]pyrazin-5-one

[0686] 5-Bromo-2-(trifluoromethyl)pyrimidine (100 mg), compound 16-6 (150 mg), and potassium carbonate (170 mg) were dissolved in N,N-dimethylformamide (2 mL). The reaction was carried out after heating to 80 °C and stirring for 10 hours. The reaction solution was cooled to room temperature, 50 mL of water was added, and the mixture was extracted with ethyl acetate (20 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by column chromatography (dichloromethane / methanol: 50 / 1; Rf: 0.6) and high performance liquid chromatography (column: YMC TAR-C18, 30 * 150 mm, 5 μm; mobile phase: [water (containing 7 mmol / L ammonium bicarbonate) - acetonitrile]; acetonitrile gradient: 70-85%) to obtain the title compound 78 (50.0 mg).

[0687] 1 H NMR (400MHz, DMSO-d6) δ8.14(s,2H),7.67(s,1H),6.61-6.14(m,1H),4.84-4.50(m,2H),3.87(brs,8H),3.48(s,3H),2.20-1.76(m,4H).

[0688] MS m / z (ESI): 485.1 [M+H] +

[0689] Example 79: 1-(2,2-difluoroethyl)-7-(7-(2-(trifluoromethyl)pyridin-4-yl)-7-azaspiro[3.5]non-2-yl)-6,7-dihydro-1H-pyrazolo[3,4-f][1,4]oxazaspiro-8(5H)-one

[0690] Step 1: Synthesis of intermediate 79-2:

[0691] Intermediate 79-1 (25.0 g) and cesium carbonate (99.4 g) were added to N,N-dimethylformamide (250 mL), followed by the slow addition of 2,2-difluoroethyl trifluoromethanesulfonate (22.3 g). The reaction was carried out under nitrogen protection and stirred at room temperature for 16 hours until completion. 300 mL of water was added to the reaction mixture, and the mixture was extracted with ethyl acetate (150 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by silica gel column chromatography (ethyl acetate / petroleum ether: 0–33%) to obtain the title compound 79-2 (12.0 g).

[0692] MS m / z(ESI): 269.0 [M+H] +

[0693] Step 2: Synthesis of intermediate 79-3:

[0694] Intermediate 79-2 (12.0 g) was added to a mixed solution of methanol / tetrahydrofuran / water (3:3:1, 20 mL), and lithium hydroxide (2.88 g) was added to the solution. The mixture was heated to 40 °C and stirred for 2 hours until the reaction was complete. The reaction solution was cooled to room temperature, and 100 mL of water was added. The mixture was extracted with ethyl acetate (50 mL * 2), and the organic phases were combined. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the title compound 79-3 (10.0 g), which was used directly in the next reaction.

[0695] MS m / z(ESI): 256.0 [M+H] +

[0696] Step 3: Synthesis of intermediate 79-5:

[0697] Intermediate 79-3 (5.00 g) and intermediate 3-1 (4.70 g) were added to N,N-dimethylformamide (100 mL), followed by N,N,N',N'-tetramethylchloroformamidin hexafluorophosphate (8.25 g) and N-methylimidazole (4.83 g). The mixture was heated to 60 °C and stirred for 16 hours until the reaction was complete. The reaction solution was cooled to room temperature, and 300 mL of water was added. The mixture was extracted with ethyl acetate (150 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (ethyl acetate / petroleum ether: 0–50%) to obtain the title compound 79-5 (3.10 g).

[0698] Step 4: Synthesis of intermediate 79-6:

[0699] Intermediate 79-5 (200 mg), 1,1'-bis(diphenylphosphine)ferrocene palladium dichloride (30.6 mg), pinacol diborate (212 mg), and potassium acetate (123 mg) were added to a 4 mL solution of 1,4-dioxane. The mixture was heated to 100 °C and stirred for 2 hours under argon protection until the reaction was complete. The reaction solution was filtered and concentrated under reduced pressure to obtain crude product 79-6 (310 mg), which was used directly in the next reaction step.

[0700] Step 5: Synthesis of intermediate 79-7:

[0701] Intermediate 79-6 (310 mg) was added to a mixed solution of tetrahydrofuran / water (6 mL / 1.5 mL), followed by sodium carbonate (125 mg) and urea peroxide (111 mg). The reaction was stirred at room temperature for 2 hours until completion. A saturated sodium thiosulfate solution (20 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (20 mL x 2). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The concentrate was then subjected to silica gel column chromatography (ethyl acetate / petroleum ether: 0%–50%) to obtain the title compound 79-7 (60.0 mg).

[0702] Step 6: Synthesis of intermediate 79-8:

[0703] Intermediate 79-7 (60.0 mg) and bromoethanol (40.0 mg) were added to N,N-dimethylformamide (1 mL), followed by potassium carbonate (60.0 mg). The mixture was heated to 50 °C and stirred for 2 hours until the reaction was complete. The reaction solution was cooled to room temperature, 10 mL of water was added, and the mixture was extracted with ethyl acetate (10 mL * 2). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 1:1) to obtain the title compound 79-8 (50.0 mg).

[0704] Step 7: Synthesis of intermediate 79-9:

[0705] Intermediate 79-8 (50.0 mg), triethylamine (22.1 mg), and methanesulfonyl chloride (18.7 mg) were added to 3 mL of dichloromethane at 0 °C. The reaction was stirred at room temperature for 1 hour until completion. 10 mL of water was added to the reaction solution, and the mixture was extracted with dichloromethane (10 mL * 2). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the title compound 79-9 (58.0 mg), which was used directly in the next reaction.

[0706] Step 8: Synthesis of intermediate 79-10:

[0707] Intermediate 79-9 (38.0 mg) was added to a solution of N,N-dimethylformamide (1 mL), followed by sodium hydride (5.00 mg, 60%). The mixture was stirred at room temperature for 16 hours until the reaction was complete. 10 mL of water was added to the reaction solution, and the mixture was extracted with ethyl acetate (10 mL x 2). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain the title compound 79-10 (31.0 mg), which was used directly in the next reaction.

[0708] Step 9: Synthesis of intermediate 79-11:

[0709] Intermediate 79-10 (46.0 mg) was added to 1 mL of dichloromethane solution, followed by 0.5 mL of 1,4-dioxane solution (4 mol / L) containing hydrogen chloride. The mixture was stirred at room temperature for 0.5 hours until the reaction was complete. The reaction solution was concentrated under reduced pressure to obtain the title compound 79-11 (35.0 mg), which was used directly in the next reaction step.

[0710] MS m / z (ESI): 341.2 [M+H] +

[0711] Step 10: Synthesis of Compound 79:

[0712] Intermediate 79-11 (20.0 mg), 4-bromo-2-(trifluoromethyl)pyridine (26.6 mg), and potassium carbonate (24.4 mg) were added to N,N-dimethylformamide (1 mL). The mixture was heated to 100 °C and stirred for 4 hours until the reaction was complete. The reaction solution was cooled to room temperature. 5 mL of water was added, and the mixture was extracted with ethyl acetate (5 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (dichloromethane / methanol: 50 / 1) and high performance liquid chromatography (column: YMC TAR-C18, 30 * 150 mm, 5 μm; mobile phase: [water (containing 7 mmol / L ammonium bicarbonate) - acetonitrile]; acetonitrile gradient: 70-85%) to obtain the title compound 79 (15.0 mg).

[0713] 1 H NMR (400MHz, DMSO-d6) δ8.23(d,J=6.3Hz,1H),7.41(s,1H),7.28(d,J=2.5Hz,1H),7.13-7.05(m,1H),6.58-5.92(m,1H),5.22-4.98(m,1H),4.92- 4.74(m,2H),4.32-4.24(m,2H),3.71-3.69(m,2H),3.54-3.48(m,2H),3. 45-3.37(m,2H),2.18-2.06(m,2H),2.00-1.86(m,2H),1.74-1.56(m,4H).

[0714] MS m / z (ESI): 486.2 [M+H] +

[0715] Examples 80-88:

[0716] The synthesis method of compound 79 was referenced, except that 4-bromo-2-(trifluoromethyl)pyridine in step 10 was replaced with the starting material in the table below, and compounds 80-88 were prepared by the same method.

[0717] Example 89: 1-(2,2-difluoroethyl)-7-(7-(6-(trifluoromethyl)pyridin-2-yl)-7-azaspiro[3.5]non-2-yl)-4,5,6,7-tetrahydropyrazolo[3,4-c]azaspiro-8(1H)-one

[0718] Step 1: Synthesis of intermediate 89-3:

[0719] Compound 79-5 (2.00 g), (E)-3-(tert-butyldimethylsiloxy)propen-1-yl-boronic acid pinacol ester (1.62 g), 1,1-bis(diphenylphosphine)ferrocene palladium dichloride (307 mg), and potassium carbonate (1.16 g) were added to a mixed solution of 1,4-dioxane (35 mL) and water (7 mL). Under nitrogen protection, the mixture was heated to 100 °C and stirred for 2 hours until the reaction was complete. The reaction solution was cooled to room temperature, 10 mL of water was added, and the mixture was extracted with ethyl acetate (50 mL * 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The concentrate was purified by silica gel column chromatography (ethyl acetate / petroleum ether: 0–50%) to obtain the title compound 89-3 (1.90 g).

[0720] Step 2: Synthesis of intermediate 89-4:

[0721] 89-3 (1.90 g) was dissolved in a mixed solution of tetrahydrofuran (38 mL) and water (19 mL), and p-toluenesulfonyl hydrazine (6.23 g) and sodium acetate (2.75 g) were added. The reaction solution was heated to 80 °C and stirred for 3 hours until the reaction was complete. The reaction solution was cooled to room temperature and extracted with ethyl acetate (30 mL * 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain crude product of the title compound 89-4 (1.80 g).

[0722] MS m / z (ESI): 571.3 [M+H] +

[0723] Step 3: Synthesis of intermediate 89-5:

[0724] The crude product 89-4 (1.80 g) obtained in the previous step was dissolved in 36 mL of tetrahydrofuran, and tetrabutylammonium fluoride (2.75 g) was added. After stirring at room temperature for 1 hour, the reaction was completed. The reaction solution was concentrated under reduced pressure, and the concentrate was purified by silica gel column chromatography (ethyl acetate / petroleum ether: 0-100%) to obtain the title compound 89-5 (600 mg).

[0725] Step 4: Synthesis of intermediate 89-6:

[0726] Compound 89-5 (600 mg) was added to 12 mL of dichloromethane, followed by triethylamine (399 mg) and then methanesulfonyl chloride (226 mg). The reaction was carried out after stirring at room temperature for 2 hours. 20 mL of water was added to the reaction mixture. The mixture was extracted with dichloromethane (10 mL x 3). The combined organic phases were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (ethyl acetate / petroleum ether: 0–80%) to obtain the title compound 89-6 (420 mg).

[0727] Step 5: Synthesis of intermediate 89-7:

[0728] Compound 89-6 (420 mg) was added to N,N-dimethylformamide (8 mL), and sodium hydride (62.9 mg, 60% purity) was added at 0 °C. The mixture was stirred at room temperature for 2 hours until the reaction was complete. 20 mL of water was added to the reaction solution, and the aqueous phase was extracted with ethyl acetate (20 mL x 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product of the title compound 89-7 (220 mg), which was directly added to the next reaction step.

[0729] MS m / z(ESI): 439.2 [M+H] +

[0730] Step 6: Synthesis of intermediate 89-8:

[0731] Intermediate 89-7 (70.0 mg) was dissolved in 1.5 mL of dichloromethane, and 0.5 mL of trifluoroacetic acid was added. The reaction was stirred at room temperature for 1 hour until the reaction was complete. The reaction solution was concentrated under reduced pressure to obtain the crude product of the title compound 89-8 (40.0 mg).

[0732] MS m / z(ESI): 339.2 [M+H] +

[0733] Step 7: Synthesis of Compound 89:

[0734] Intermediate 89-8 (40.0 mg) and 2-bromo-6-(trifluoromethyl)pyridine (32.1 mg) were dissolved in N,N-dimethylformamide (1 mL), potassium carbonate (81.6 mg) was added, and the mixture was heated to 100 °C and stirred for 2 hours until the reaction was complete. The reaction solution was cooled to room temperature, 5 mL of water was added, and the aqueous phase was extracted with ethyl acetate (10 mL * 3). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by high performance liquid chromatography (column: YMC TAR-C18, 30 * 150 mm, 5 μm; mobile phase: [water (containing 7 mmol / L ammonium bicarbonate) - acetonitrile]; acetonitrile gradient: 70-85%) to obtain the title compound 89 (29.7 mg).

[0735] 1 H NMR (400MHz, DMSO-d6) δ7.72(t,J=8.0Hz,1H),7.48(s,1H),7.11(d,J=8.9Hz,1H),7.00(d,J=7.2Hz,1H),6.44-6.16(m,1 H),4.89-4.78(m,3H),3.66-3.22(m,6H),2.69(t,J=7.3Hz,2H),2.23-2.18(m,2H),2.08-1.85(m,4H),1.70-1.57(m,4H).

[0736] MS m / z (ESI): 484.3 [M+H] +

[0737] Examples 90-97:

[0738] The synthesis method of compound 89 was referenced, except that 2-bromo-6-(trifluoromethyl)pyridine in step 7 was replaced with the starting material in the table below, and compounds 90-97 were prepared by the same method.

[0739] Example 98: (R)-1-(2,2-difluoroethyl)-4-methyl-6-((2-(2-(trifluoromethyl)pyrimidin-5-yl)-2-azaspiro[3.4]oct-6-yl)oxy)-1,4-dihydro-5H-pyrazolo[3,4-b]pyrazin-5-one

[0740] The title compound 98 was prepared by referring to the synthesis of Example 61, except that 61-1 in step 1 was replaced with 98-1.

[0741] 1H NMR(400MHz,MeOD-d4)δ8.04(s,2H),7.71(s,1H),6.40-6.09(m,1H),5.59-5.51(m,1H), 4.73-4.63(m,2H),4.14-4.02(m,4H),3.64(s,3H),2.46-2.22(m,4H),2.12-1.98(m,2H).

[0742] MS m / z (ESI): 486.2 [M+H] +

[0743] Example 99: (S)-1-(2,2-difluoroethyl)-4-methyl-6-((2-(2-(trifluoromethyl)pyrimidin-5-yl)-2-azaspiro[3.4]oct-6-yl)oxy)-1,4-dihydro-5H-pyrazolo[3,4-b]pyrazin-5-one

[0744] The title compound 99 was prepared by referring to the synthesis of Example 61, except that 61-1 in step 1 was replaced with 99-1.

[0745] 1 H NMR(400MHz,MeOD-d4)δ8.04(s,2H),7.71(s,1H),6.24(tt,J=55.2,4.0Hz,1H),5.60-5.51(m,1H), 4.68(td,J=14.1,4.0Hz,2H),4.17-4.01(m,4H),3.64(s,3H),2.46-2.23(m,4H),2.11-1.99(m,2H).

[0746] MS m / z (ESI): 486.2 [M+H] +

[0747] Example 100: 1-(2,2-difluoroethyl)-7-(2-(6-(trifluoromethyl)pyridin-2-yl)-2-azaspiro[3.5]non-7-yl)-4,5,6,7-tetrahydropyrazolo[3,4-c]azaspiro-8(1H)-one

[0748] Step 1 follows the same synthesis method as Step 3 of Example 79, except that starting material 3-1 is replaced with 100-1, and title compound 100-2 is prepared in the same manner.

[0749] Steps 2 to 8 are synthesized using the same method as in Example 89, except that starting material 79-5 is replaced with 100-2, and title compound 100 (15.0 mg) is obtained by the same method.

[0750] 1 H NMR (400MHz, DMSO-d6) δ7.70(t,J=7.8Hz,1H),7.45(s,1H),7.00(d,J=7.3Hz,1H),6.61(d,J=8.4Hz,1H),6.46-6.16(m,1H),4.88-4. 75(m,2H),4.27(brs,1H),3.81(s,2H),3.67(s,2H),3.22(t,J=5.9Hz,2H),2.67-2.63(m,2H),2.06-1.83(m,4H),1.67-1.55(m,6H).

[0751] MS m / z(ESI): 483.9 [M+H] +

[0752] Examples 101-108:

[0753] The synthesis method of compound 100 was referenced, except that 2-bromo-6-(trifluoromethyl)pyridine in step 8 was replaced with the starting material in the table below, and compounds 101-108 were prepared by the same method.

[0754] Example 109: 1-(2,2-difluoroethyl)-7-(2-(2-(trifluoromethyl)pyridin-4-yl)-2-azaspiro[3.5]non-7-yl)-6,7-dihydro-1H-pyrazolo[3,4-f][1,4]oxazaspiro-8(5H)-one

[0755] Steps 1 to 7 were performed using the same synthesis method as in Example 79, except that starting material 79-5 was replaced with 100-2, and title compound 109 (20.0 mg) was obtained in the same manner.

[0756] 1 H NMR (400MHz, DMSO-d6) δ8.23(d,J=5.7Hz,1H),7.41(s,1H),6.70(d,J=1.7Hz,1H),6.55-6.49(m,1H),6.32(tt,J=56.2,4.1Hz,1H),4.84(td, J=14.2,4.1Hz,2H),4.47-4.37(m,1H),4.26-4.24(m,2H),3.80(s,2H) ,3.69(s,2H),3.59-3.57(m,2H),2.01-1.98(m,2H),1.74-1.45(m,6H).

[0757] MS m / z (ESI): 486.2 [M+H] +

[0758] Examples 110-118:

[0759] The synthesis method of compound 109 was referenced, except that 4-bromo-2-(trifluoromethyl)pyridine in step 7 was replaced with the starting material in the table below, and compounds 110-118 were prepared by the same method.

[0760] Example 119: 4-Methyl-1-(2,2,2-trifluoroethyl)-6-(2-(2-(trifluoromethyl)pyridin-4-yl)-2,7-diazaspiro[3.5]non-7-yl)-1,4-dihydro-5H-pyrazolo[4,3-b]pyridin-5-one

[0761] The title compound 119 was prepared by referring to the synthetic method of Example 57 and the synthetic route described above.

[0762] 1 H NMR (400MHz, DMSO-d6) δ8.23(d,J=5.7Hz,1H),7.82(s,1H),7.35(s,1H),6.74(d,J=2.2Hz,1H),6.56( dd,J=5.7,2.2Hz,1H),5.39-5.29(m,2H),3.80(m,4H),3.52(s,3H),3.11(brs,4H),1.96-1.87(m,4H).

[0763] MS m / z (ESI): 501.2 [M+H] +

[0764] Experimental Example 1: Imaging determination of the high lysosomal GCase activity of the disclosed compound in the SH-SY5Y cell line.

[0765] 1) Materials and reagents:

[0766] 2) Instruments:

[0767] 3) Experimental methods:

[0768] Complete cell culture medium: DMEM / F-12, HEPES, containing 10% FBS and 1% penicillin-streptomycin; High content imaging detection medium: DMEM / F-12, HEPES, phenol red-free and FBS-free;

[0769] 20,000 SH-SY5Y cells were seeded per well in a 384-well plate, along with 40 μL of complete cell culture medium per well, and incubated overnight at 37°C with 5% carbon dioxide.

[0770] Dissolve the disclosed compound in DMSO to a concentration of 20 mM, and then dilute it 1:1 with DMSO to 10 mM. The 20 mM and 10 mM compound solutions were then diluted 1:400 with complete cell culture medium to obtain concentrations of 50 μM and 25 μM, respectively. Add 10 μL of culture medium containing 50 μM or 25 μM of the disclosed compound to the wells of a 384-well plate, resulting in final concentrations of the test compound of 10 μM or 5 μM, and a final DMSO concentration of 0.05%. Centrifuge at 500 rpm for 30 seconds and incubate at 37°C with 5% CO2 for 72 hours.

[0771] Prepare high-content imaging detection medium (DMEM / F-12, HEPES, phenol red-free, FBS-free), add 75 nM Lysotracker and 58 μM PFB-FDGlu to a final concentration; remove the original supernatant from the cell plate, add medium containing 40 μL Lysotracker and PFB-FDGlu, centrifuge at 500 rpm for 30 s, and incubate at 37°C and 5% CO2 for 45 minutes; remove the cell plate and perform continuous high-content imaging at 30-minute intervals for a total duration of 3 hours, acquiring images from brightfield, Alexa 488, and Alexa 647 channels, with 10 fields of view per well. Calculate the average fluorescence intensity of cells, lysosomes, and cell-lysosome colocalization. Using the untreated DMSO wells as a background, calculate the fluorescence intensity at the endpoint of each well. The drug concentration at which the fluorescence intensity of the analyte is twice the background fluorescence intensity is taken as the AC concentration. 100 Value (AC) 100 This refers to the drug concentration at which the GCase activation activity is 200%.

[0772] The disclosed compound exhibited high GCase activation levels in SH-SY5Y cells, as shown in Table 1. (Compound AC was also tested.) 100 Value range: A: AC 100 ≤5μM; B: 5uM <AC 100 ≤10μM; C: >10μM;

[0773] Table 1. Results of the assay of GCase activity of the disclosed compounds in SH-SY5Y cells.

[0774] Experimental Example 2: Inhibitory effect of the compounds of the present invention on the activities of CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4 enzymes.

[0775] Unless otherwise specified, all reagents and consumables used in this experiment are commercially available products.

[0776] The inhibition of CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4 enzyme activities by the compounds of this invention was determined using the following experimental method.

[0777] The experimental steps are as follows:

[0778] 1. Preparation of 100mM Phosphate Buffered Sodium (PBS): Weigh 7.098g Na₂HPO₄ and dissolve it in 500mL of pure water by sonication to obtain solution A. Weigh 3.400g KH₂PO₄ and dissolve it in 250mL of pure water by sonication to obtain solution B. Slowly add solution B to solution A on a stirrer until the pH reaches 7.4 to prepare a 100mM PBS buffer.

[0779] 2. Preparation of NADPH solution: Prepare 10mM NADPH (reduced coenzyme II) solution using 100mM PBS buffer.

[0780] 3. Preparation of working solutions for the test compounds: Dilute 10 mM of the DMSO stock solution of the compounds of this invention with DMSO to obtain working solutions of the compounds (10000 μM, 3333.3 μM, 1111.1 μM, 370.37 μM, 123.46 μM, 41.15 μM, 13.7 μM, 0 μM).

[0781] 4. Preparation of positive inhibitor working solution: Dilute the positive inhibitor DMSO stock solution with DMSO (CYP 2C9: sulfamethoxazole 1000μM, CYP 2D6: quinidine / CYP 3A4: ketoconazole, 100μM; CYP 1A2: α-naphthylflavonoid, 50μM; CYP 2C19: Nocacolone, 6000 μM) yielded positive inhibitor working solutions (sulfamethoxazole, 1000 μM, 300 μM, 100 μM, 30 μM, 10 μM, 3 μM, 0 μM; quinidine / ketoconazole, 100 μM, 30 μM, 10 μM, 3 μM, 1 μM, 0.3 μM, 0 μM; α-naphthylflavonoid, 50 μM, 15 μM, 5 μM, 1.5 μM, 0.5 μM, 0.15 μM, 0 μM; nocacolone, 6000 μM, 1800 μM, 600 μM, 180 μM, 60 μM, 18 μM, 0 μM).

[0782] 5. Preparation of substrate working solution: Prepare substrate working solution (8000 μM phenacetin, 1000 μM diclofenac, 8000 μM S-methoxytocin, 1000 μM dextromethorphan and 400 μM midazolam) with water, acetonitrile or acetonitrile / methanol.

[0783] 6. Take 1 μL of 20 mg / mL liver microsome solution (source: gentest), 1 μL of substrate working solution, and 177 μL of PBS buffer, add 1 μL of compound working solution (for the positive control group, add 1 μL of positive inhibitor working solution instead of the compound working solution), mix well, and pre-incubate in a 37°C water bath for 10 minutes. Add 10 mM NADPH solution and pre-incubate in a 37°C water bath for 10 minutes. After 10 minutes, add 20 μL of NADPH to each well to start the reaction. Incubate at 37°C for 15 minutes (CYP1A2), 15 minutes (CYP2C9), 60 minutes (CYP2C19), 15 minutes (CYP2D6), or 15 minutes (CYP3A4). All incubated samples should be in duplicate. After the corresponding incubation time, add 400 μL of ice-cold methanol containing the internal standard to all samples to terminate the reaction. Vortex to mix, and centrifuge at 4000 rpm and 4°C for 30 minutes. After centrifugation, transfer 100 μL of supernatant to the sample plate, add 200 μL of ultrapure water and mix well for LC-MS / MS analysis.

[0784] The IC50 values ​​of the compound of this invention against CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4 were calculated using Excel XLfit 5.3.1.3. 50 The values ​​are shown in Table 2.

[0785] Table 2 shows the IC50 values ​​of the compounds of the present invention against CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4. 50 value "-" indicates that no detection was performed.

[0786] Experimental Example 3: Determination of the potential inhibitory effect of the compounds of this invention on the voltage-gated potassium ion channel hERG

[0787] This experiment used the Chinese hamster ovary (CHO) cell line (B'SYS GmbH) stably expressing the hERG potassium channel. hERG-CHO cells were clamped in a whole-cell voltage-clamp configuration using a SyncroPatch 384i / 384 (Nanion) automated patch-clamp system, and hERG currents were induced by appropriate voltages. The test compounds were then tested: cells were first perfused six times with extracellular fluid containing 0.1% DMSO. The measured stable hERG current was used as the baseline, and the baseline current value was the mean of five stable sampling points (tail current magnitude). 空白After the hERG current stabilized, five concentration gradients of the test compound (0.37 μM, 1.11 μM, 3.33 μM, 10 μM, and 30 μM) were perfused around the cells. The cells were allowed to fully react with the compound for 10 minutes while the hERG current was recorded simultaneously. Once the current stabilized, five stable hERG current values ​​were read, and their average was taken as the final current value at the specific concentration (tail current magnitude). 化合物 Cisapride (Sigma (C4740)) was used as a positive control for simultaneous assays at six concentrations (0.001 μM, 0.004 μM, 0.012 μM, 0.037 μM, 0.111 μM, and 0.333 μM) to verify the stability of the test cells and the accuracy of the results. After testing the compounds, 450 nM of Dopiride (Beijing Yipulis Technology Development Co., Ltd., D525700) was added to all test cells to completely suppress their current, serving as a complete positive control for these cells (tail current size). 阳性对照 Finally, the tail current suppression rate is calculated using the following formula: {Tail current suppression rate (%) = [1 - (tail current magnitude)]} 化合物 - Tail current magnitude 阳性对照 ) / (tail current magnitude) 空白 - Tail current magnitude 阳性对照 The dose-response curve was then fitted using Graphpad Prism 8.0 software, and the IC was calculated. 50 value.

[0788] The inhibition of hERG by the disclosed compounds is shown in Table 3 below. The test data indicate that the hERG inhibitory activity of the disclosed compounds is poor, and the potential safety risks in the future are relatively small.

[0789] Table 3. Inhibitory activity of the disclosed compounds in the voltage-gated potassium channel hERG.

[0790] Experimental Example 4: Flow Cytometry Determination of the Effect of the Disclosed Compounds on GCase Enzyme Activity in Human Fibroblasts

[0791] 1) Materials and reagents

[0792] 2) Instruments

[0793] 3) Experimental methods:

[0794] 10,000 fibroblasts were seeded into each well of a 96-well plate with 100 μL of complete cell culture medium (DMEM containing 10% FBS and 1% penicillin-streptomycin) and incubated at 37°C in a 5% CO2 incubator for 24 hours.

[0795] Dissolve the disclosed compound in DMSO to 100 mM, and then dilute it to 0.08-100 mM with DMSO as needed. Dilute the diluted DMSO solution of the compound with complete cell culture medium at a ratio of 1:1000 to a concentration of 0.08-100 μM. Add 100 μL of culture medium containing the disclosed compound to the wells of a 96-well plate to make a working concentration of 0.04-50 μM and a final DMSO concentration of 0.05%. Incubate at 37°C in a 5% CO2 incubator for 72 hours.

[0796] Lysosomal GCase in fibroblasts can specifically react with the substrate 5-(pentafluorobenzoylamino)fluorescein-di-β-D-glucopyranoside (PFB-FDGlu) to generate a fluorescent signal. Before the assay, 2 μL of 100 mM MCBE solution was added to the background control wells and the cells were blocked for 1 hour. After blocking, 1.155 mM PFB-FDGlu was added to the complete culture medium to prepare the chromogenic reaction solution. 22 μL of the chromogenic reaction solution was added to each well to bring the final concentration of PFB-FDGlu to 115.5 μM, and the cells were incubated at 37°C in a 5% CO2 incubator in the dark for 2.5 hours. After incubation, the supernatant was discarded, and the cells were washed twice with 100 μL of phenol red-free DMEM medium. 75 μL of 0.25% trypsin (containing EDTA) was added to each well for digestion for 5 minutes, followed by the addition of 75 μL of phenol red-free DMEM containing 10% FBS to terminate the digestion. After digestion, transfer the cells to a 96-well plate, centrifuge at 300g for 5 minutes, discard the supernatant, and resuspend the cells thoroughly in 200μL of PBS containing 1% FBS.

[0797] The fluorescence signal of cells in each well was detected by flow cytometry. The detection channels were set according to the excitation / emission wavelengths of the PFB-FDGlu fluorescent dye Ex / Em = 494 / 521 nm. The forward scattering (FSC) and side scattering (SSC) voltages were adjusted, and debris and cell clumps were excluded. At least 8000 cells were collected from each sample well. The PFB-FDGlu channel signals were analyzed using FlowJo software, and the average fluorescence intensity of cells in each well was calculated.

[0798] After subtracting the background signal read from the CBE inhibition group (background control well), the fluorescence intensity of the untreated DMSO well (blank control well) was used as the baseline value to calculate the percentage of GCase activation in the test compound well relative to the blank control well.

[0799] The activation curve was statistically analyzed and plotted using Prism software. A fitting function was used, with the drug working concentration on the horizontal axis and the percentage of activation relative to DMSO on the vertical axis, to calculate the AC. 50 Value. Among them, AC 50 This refers to the drug concentration at which the GCase enzyme activation percentage is 150%.

[0800] The disclosed compound exhibited high GCase activation levels in human fibroblasts, as shown in Table 4.

[0801] Table 4. Flow cytometry analysis of the effects of the disclosed compounds on GCase activity in human fibroblasts.

[0802] Experimental Example 5: Detection of the pharmacokinetic properties of the compounds of this invention in mice

[0803] Experimental materials

[0804] 1. Healthy adult male C57BL / 6 mice were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0805] 2. DMSO, PBS (phosphate buffer), Solutol HS15, NMP (N-methylpyrrolidone), HP-β-CD (hydroxypropyl-β-cyclodextrin), Transcutol P (diethylene glycol monoethyl ether), hydrochloric acid, tolbutamide, acetonitrile, and formic acid were purchased from Merck (USA).

[0806] 3. K2EDTA anticoagulant tubing was purchased from Jiangsu Xinkang Medical Equipment Co., Ltd.

[0807] II. Experimental Methods

[0808] 1. Drug preparation: Weigh a certain amount of the compound disclosed herein, dissolve it in 5% NMP + 5% 1M hydrochloric acid + 30% Transcutol P + 60% (40% HP-β-CD) to prepare a 0.3 mg / mL solution for oral administration; weigh a certain amount of the compound disclosed herein, dissolve it in 5% DMSO + 10% solubil HS15 + 85% PBS to prepare a 0.2 mg / mL solution for intravenous administration.

[0809] 2. Administration methods: Gavage group: C57BL / 6 mice were administered the drug via gavage without fasting, at a dose of 3 mg / kg. Intravenous group: C57BL / 6 mice were administered the drug intravenously without fasting, at a dose of 1 mg / kg.

[0810] 3. Animal experiments: After oral or intravenous administration to mice, blood samples of more than 20 μL were collected from the orbital cavity at 0.083 (intravenous administration only), 0.25, 0.5, 1, 2, 4, 8, 24, 48 and 72 hours after administration. The blood was added to K2EDTA anticoagulant tubes, centrifuged at 12000 rpm, 4℃ for 5 minutes to separate the plasma, and stored at -90 to -60℃.

[0811] 4. Determination of the content of the analyte compound in mouse plasma after gavage or intravenous administration of different concentrations of the drug: Melt the sample at room temperature and vortex for 1 min; quantitatively transfer 10 μL to a 2 mL 96-well plate, add 100 μL of precipitant (containing 10 ng / mL tolbutamide acetonitrile solution), and shake (1000 rpm × 3 min); centrifuge (4000 rpm × 15 min), transfer 80 μL of the supernatant to a 2 mL 96-well plate, add 80 μL of diluent (water), and shake well (1000 rpm × 3 min). Quantitative detection was performed using an LC-MS / MS system (AB Sciex Triple Quad 5500+).

[0812] 5. Data Processing: Pharmacokinetic parameters were calculated using the non-compartmental model statistical moment method with Phoenix WinNonlin 8.0 software (Certara, USA). The experimental results are shown in Tables 5 and 6. The test data indicate that the compound disclosed herein has good oral absorption in mice.

[0813] Table 5. Pharmacokinetic parameters of the compound after a single oral gavage administration to mice.

[0814] Table 6 Pharmacokinetic parameters of the compound after a single intravenous administration in mice.

[0815] Experimental Example 6: Detection of the pharmacokinetic properties of the compounds of the present invention in rats

[0816] Experimental materials

[0817] 1. Healthy adult male SD rats were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0818] 2. DMSO, PBS (phosphate buffer), Solutol HS15, NMP (N-methylpyrrolidone), HP-β-CD (hydroxypropyl-β-cyclodextrin), Transcutol P (diethylene glycol monoethyl ether), hydrochloric acid, tolbutamide, acetonitrile, and formic acid were purchased from Merck (USA).

[0819] 3. K2EDTA anticoagulant tubing was purchased from Jiangsu Xinkang Medical Equipment Co., Ltd.

[0820] II. Experimental Methods

[0821] 1. Drug preparation: Weigh a certain amount of the compound disclosed herein, dissolve it in 5% NMP + 5% 1M hydrochloric acid + 30% Transcutol P + 60% (40% HP-β-CD) to prepare a 0.3 mg / mL solution for oral administration; weigh a certain amount of the compound disclosed herein, dissolve it in 5% DMSO + 10% solubil HS15 + 85% PBS to prepare a 0.2 mg / mL solution for intravenous administration.

[0822] 2. Administration methods: Gavage group: SD rats were administered the drug via gavage after fasting, with a dosage of 3 mg / kg. Intravenous group: SD rats were administered the drug intravenously after fasting, with a dosage of 1 mg / kg.

[0823] 3. Animal experiments: After oral or intravenous administration to rats, 100 μL of blood was collected from the jugular vein at 0.083 (intravenous administration only), 0.25, 0.5, 1, 2, 4, 8, 24, 48 and 72 hours after administration and added to an anticoagulant tube. The plasma was separated by centrifugation at 12000 rpm, 4℃ for 5 minutes and stored at -90 to -60℃.

[0824] 4. Determination of the content of the target compound in rat plasma after gavage or intravenous administration of different concentrations of the drug: The sample was thawed at room temperature and vortexed for 1 min; 10 μL was quantitatively transferred to a 2 mL 96-well plate, 100 μL of precipitant (containing 10 ng / mL tolbutamide acetonitrile solution) was added, and the plate was shaken (1000 rpm × 3 min); centrifuged (4000 rpm × 15 min), and 80 μL of the supernatant was transferred to a 2 mL 96-well plate, 80 μL of diluent (water) was added, and the plate was shaken well (1000 rpm × 3 min). Quantitative detection was performed using an LC-MS / MS system (AB Sciex Triple Quad 5500+).

[0825] 5. Data Processing: Pharmacokinetic parameters were calculated using the non-compartmental model statistical moment method with Phoenix WinNonlin 8.0 software (Certara, USA). The experimental results are shown in Tables 7 and 8. The test data indicate that the compound disclosed herein has good oral absorption in rats.

[0826] Table 7 Pharmacokinetic parameters of the compound after a single oral gavage administration to rats

[0827] Table 8. Pharmacokinetic parameters of the compound after a single intravenous administration in rats.

[0828] Experimental Example 7: Detection of the pharmacokinetic properties of the compound of the present invention in dogs

[0829] Experimental materials

[0830] 1. A healthy adult male Beagle was purchased from Jiangsu Mas Biotechnology Co., Ltd.

[0831] 2. DMSO, PBS (phosphate buffer), Solutol HS15, CMC-Na (sodium carboxymethyl cellulose), tolbutamide, acetonitrile, and formic acid were purchased from Merck (USA).

[0832] 3. K2EDTA anticoagulant tubing was purchased from Jiangsu Xinkang Medical Equipment Co., Ltd.

[0833] II. Experimental Methods

[0834] 1. Drug preparation: Weigh a certain amount of the disclosed compound, dissolve it in 0.5% CMC-Na to prepare a 0.6 mg / mL solution, and administer by gavage. Weigh a certain amount of the disclosed compound, dissolve it in 5% DMSO + 10% Solutol HS15 + 85% PBS to prepare a 0.5 mg / mL solution, and administer by intravenous injection.

[0835] 2. Administration method: Gavage group: The dosage was 3 mg / kg. Intravenous group: The dosage was 1 mg / kg.

[0836] 3. Animal experiments: After administration by gavage or intravenous in beagle dogs, 1000 μL of blood was collected from the forelimb vein at 0.083 (intravenous administration only), 0.25, 0.5, 1, 2, 4, 8, 24, 48 and 72 hours after administration and added to a K2EDTA anticoagulant tube. The plasma was separated by centrifugation at 12000 rpm, 4°C for 5 minutes and stored at -90 to -60°C.

[0837] 4. Determination of the content of the analyte compound in canine plasma after oral or intravenous administration of different concentrations of the drug: Melt the sample at room temperature and vortex for 1 min; quantitatively transfer 20 μL to a 2 mL 96-well plate, add 200 μL of precipitant (containing 10 ng / mL tolbutamide acetonitrile solution), shake (1000 rpm × 3 min), centrifuge (4000 rpm × 15 min), transfer 80 μL of the supernatant to a 2 mL 96-well plate, add 80 μL of diluent (water), and shake well (1000 rpm × 3 min). Quantitative detection was performed using an LC-MS / MS system (AB Sciex Triple Quad 5500+).

[0838] 5. Data Processing: Pharmacokinetic parameters were calculated using the statistical moment method of non-compartmental models in Phoenix WinNonlin 8.0 software (Certara, USA). The experimental results are shown in Tables 9 and 10.

[0839] Table 9. Pharmacokinetic parameters of the compound after a single oral gavage administration in dogs.

[0840] Table 10 Pharmacokinetic parameters of the compound after a single intravenous administration in dogs

[0841] Experimental Example 8: Detection of the rat brain penetration properties of the compounds of this invention

[0842] Experimental materials

[0843] 1. Healthy adult male SD rats were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0844] 2. NMP (N-methylpyrrolidone), HP-β-CD (hydroxypropyl-β-cyclodextrin), Transcutol P (diethylene glycol monoethyl ether), hydrochloric acid, tolbutamide, acetonitrile, and formic acid were purchased from Merck (USA).

[0845] 3. K2EDTA anticoagulant tubing was purchased from Jiangsu Xinkang Medical Equipment Co., Ltd.

[0846] II. Experimental Methods

[0847] 1. Drug preparation: Weigh a certain amount of the disclosed compound, dissolve it in 5% NMP + 5% 1M hydrochloric acid + 30% Transcutol P + 60% (40% HP-β-CD) to prepare a 0.3 mg / mL solution for oral administration.

[0848] 2. Administration method: SD rats were administered the drug via gavage after fasting.

[0849] 3. Animal Experiments: Plasma, brain tissue, and cerebrospinal fluid (CSF) were collected from rats at 0.25, 2, 8, 24, and 72 hours after administration. Plasma collection: 100 μL of blood was collected from the jugular vein and added to an anticoagulant tube. The plasma was separated by centrifugation at 12,000 rpm, 4°C, for 5 minutes and stored at -90 to -60°C. CSF collection: Rats were euthanized using the carbon dioxide method. ~50 μL of CSF was collected by puncture. A 1 mL syringe was used, with the needle bevel facing upwards and the needle tip nearly horizontally inserted into the subarachnoid space. The needle was fixed, and CSF was slowly aspirated. The collected samples were stored at -90 to -60°C. Before CSF measurement, acetonitrile (CSF:acetonitrile = 1:1, v / v) was added to the collection tube for mixing. Brain tissue collection: Cardiac perfusion was performed first, followed by brain tissue harvesting. Under deep terminal anesthesia, the heart was perfused with pre-cooled saline solution to flush out any remaining blood from the brain tissue. After brain tissue was harvested, it was gently washed once with frozen physiological saline, blotted dry, weighed, and aliquoted into homogenization tubes (approximately 100–300 mg per tube). The tubes were then transferred to a -90–-60°C environment for cryopreservation. Before brain tissue analysis, PBS (brain:PBS = 1:4, w / v) was added for homogenization.

[0850] 4. Determination of the content of the target compound in rat plasma, cerebrospinal fluid, and brain tissue after gavage administration of the drug: The sample was thawed at room temperature and vortexed for 1 min; 10 μL was quantitatively transferred to a 2 mL 96-well plate, 100 μL of precipitant (containing 10 ng / mL tolbutamide acetonitrile solution) was added, and the plate was shaken (1000 rpm × 3 min); the plate was centrifuged (4000 rpm × 15 min), and 80 μL of the supernatant was transferred to a 2 mL 96-well plate, 80 μL of diluent (water) was added, and the plate was shaken well (1000 rpm × 3 min). Quantitative detection was performed using an LC-MS / MS system (AB Sciex Triple Quad 5500+).

[0851] 5. Data Processing: Pharmacokinetic parameters were calculated using the non-compartmental model statistical moment method with Phoenix WinNonlin 8.0 software (Certara, USA). The experimental results are shown in Table 11. The test data indicate that the compound disclosed herein has good brain penetration properties in rats.

[0852] Table 11 Brain transillumination data of rats after a single gavage administration of the compound disclosed herein

Claims

A compound represented by Formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, in, Ring C is selected from the following structural groups: Indicates a single bond or a double bond; U is selected from C=0, N and CR 2 ; Q is selected from C and N; V is selected from O, S, NR 3 and CHR 2 ; t' is 1 or 2; s' is selected from 0, 1, or 2; R 5 selected from halogen, =0, Ci-C6-alkyl, Ci-C6-haloalkyl or Ci-C4-alkoxy; R c selected from C1-C6alkyl, C1-C6alkoxy, C3-C6cycloalkyl, or 4-6 membered heterocyclyl, each independently optionally substituted with NH2, OH, halogen, or C1-C6alkyl; R 2 selected from H, halogen, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C4 alkoxy; R 3 selected from H, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C4 alkoxy; L, W are each independently selected from the group consisting of a bond, -CR a R b -, C(O), O, S, -CR a R b O- or NR a ; R a , R b is independently selected from H, halogen, Ci-C6alkyl, Ci-C6haloalkyl or Ci-C4alkoxy; Ring B is selected from the following groups: a)C6-C 14 Spirocycloalkylene, C6-C 14 Dyscyclic alkylene, C5-C 12 Bridged cycloalkyl groups, 6-14 membered spirocycloalkyl groups, 6-14 membered fused heterocyclic groups, or 5-12 membered bridged heterocyclic groups; or, b) Each R 4 It is independently selected from deuterium, OH, halogen, =O, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy or C3-C6 cycloalkyl; Cycloalkyl, 4- to 12-membered heterocyclyl, 5- to 10-membered heteroaryl, or C6-C10aryl; 12 Cycloalkyl, 4- to 12-membered heterocyclyl, 5- to 10-membered heteroaryl, or C6-C10aryl; 10 Cycloalkyl, Each R 1 The group is independently selected from CN, NH2, OH, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C3-C6 cycloalkyl or 4-10 heterocyclic group, wherein each of the C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl or 4-10 heterocyclic group is independently and optionally substituted by NH2, OH, halogen or C1-C6 alkyl; n is selected from 0, 1, or 2; p and q are independently selected from 0, 1, or 2; provided that when ring C is selected from at the time, L is selected from -CR a R b -, C(O), O or S. The compound of Formula (I) according to claim 1, or a stereoisomer or a pharmaceutically acceptable salt thereof, wherein, The compound of formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof is selected from the group consisting of a compound of formula (I-1) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein U is selected from N and CR 2 ; Ring A, Ring B, W, L, V, R 1 , R 4 , R c , n, p, q are defined as in claim 1. The compound of Formula (I) according to claim 1 or 2, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein, The compound of formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof is selected from the group consisting of a compound of formula (I-1B) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein ring A, ring B, W, L, R 1 , R 2 , R 3 , R 4 , R c , p, q, n are as defined in claim 1. The compound of Formula (I) according to any one of claims 1-3, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein, The compound of formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof is selected from the group consisting of a compound of formula (I-1B-1) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein ring A, ring B, W, L, R 1 , R 2 , R 3 , R 4 , R c , p, q are as defined in claim 1. The compound of Formula (I) according to claim 1, or a stereoisomer or a pharmaceutically acceptable salt thereof, wherein, The compound according to Formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof is selected from the group consisting of a compound according to Formula (I-2) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein ring A, ring B, W, L, V, R 1 , R 4 , R 5 , R c , n, p, q, t', s' are as defined in claim 1. The compound of Formula (I) according to claim 1 or 5, or a stereoisomer or a pharmaceutically acceptable salt thereof, wherein, The compound of formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof is selected from a compound according to formula (I-2A) or (I-2B) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein ring A, ring B, W, L, V, R 1 , R 4 , R c , n, p, q are as defined in claim 1. The compound of Formula (I) or a stereoisomer or pharmaceutically acceptable salt thereof according to any one of claims 1-2, wherein, U is selected from C=O, N, and CH; Alternatively, U can be N; or U is CR 2 . The compound of Formula (I) or a stereoisomer or pharmaceutically acceptable salt thereof according to any one of claims 1-2 and 5-6, wherein, V is selected from O, NR 3 and CHR 2 ; Alternatively, V is selected from NR 3 or CHR 2 ; Alternatively, V is selected from O or CHR 2 ; Alternatively, V can be selected from O or CH2; Alternatively, V is NR 3 . The compound of Formula (I) according to claim 1 or 5, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein, R 5 =O; s' is 0 or 1. The compound of Formula (I) according to any one of claims 1-9, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein, Ring C is selected from the following structural groups: Alternatively, ring C is selected from the following structural groups: Alternatively, ring C is selected from the following structural groups: Alternatively, ring C is selected from the following structural groups: Alternatively, ring C is selected from the following structural groups: Alternatively, ring C is selected from the following structural groups: Alternatively, ring C is selected from the following structural groups: The compound of Formula (I) according to any one of claims 1-10, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein, R c selected from C1-C6alkyl or 4-6 membered heterocyclyl, each independently optionally substituted with NH2, OH, halogen, or C1-C6alkyl; Or, R c Selected from C1-C6 alkyl groups, wherein the C1-C6 alkyl groups are optionally substituted with NH2, OH, halogens or C1-C6 alkyl groups; Or, R c Selected from C1-C6 alkyl groups, wherein the C1-C6 alkyl groups are optionally substituted with halogens; or R c is selected from CH3, CH2CH3, CH2CHF2, CH2CF2CH3, CH2CF3, or oxetanyl. The compound of Formula (I) according to any one of claims 1-11, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein, R 2 selected from H or Ci-C6alkyl; or R 3 is selected from H or C1-C6 alkyl; or, R 3 is selected from H or CH3. The compound of Formula (I) or a stereoisomer or pharmaceutically acceptable salt thereof according to any one of claims 1-12, wherein, L, W are each independently selected from a bond, -CR a R b -, C(O), O, S, or CH2O; or each L, W is independently selected from a bond, -CR a R b -, C(O), O, or S; Alternatively, L is a bond and W is selected from a bond, -CR a R b -, C(O), O or S; or L is selected from a bond, -CR a R b -, O, CH2O or NR a , and W is a bond; Alternatively, L is selected from bond, -CH2-, O, CH2O or NH, and W is bond; Alternatively, L is selected from a bond, -CR a R b -, C(O), O or S, and W is a bond; Alternatively, L is -CH2- and W is selected from a bond, -CR a R b -, C(O), O or S; or L is a bond, -CR a R b -, C(O), O or S, and W is a bond; Alternatively, L is selected from bond, -CH2- or O, and W is selected from bond or CH2O; Alternatively, L is a bond, and W is CH2O; Alternatively, L is O and W is a bond; Alternatively, L is -CH2- and W is a bond; Alternatively, L is the key and W is the key. The compound of formula (I) according to any one of claims 1-13, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, Ring B is selected from 6-14 membered spiro-heterocyclic groups, 6-14 membered fused-heterocyclic groups, or 5-12 membered bridged-heterocyclic groups; Alternatively, ring B is selected from 6-14 membered bicyclic spirocycloheterocyclic groups; Alternatively, ring B is selected from 3-membered / 6-membered bicyclic spiro-heterocyclic base, 4-membered / 4-membered bicyclic spiro-heterocyclic base, 4-membered / 5-membered bicyclic spiro-heterocyclic base, 4-membered / 6-membered bicyclic spiro-heterocyclic base, 5-membered / 5-membered bicyclic spiro-heterocyclic base or 5-membered / 6-membered bicyclic spiro-heterocyclic base; Alternatively, ring B is selected from 4-membered / 4-membered bicyclic spiro-heterocyclic base, 4-membered / 5-membered bicyclic spiro-heterocyclic base, 5-membered / 6-membered bicyclic spiro-heterocyclic base or 4-membered / 6-membered bicyclic spiro-heterocyclic base; Alternatively, ring B is selected from 6-14 membered bicyclic fused heterocyclic groups; Alternatively, ring B is selected from 3-membered / 5-membered bicyclic fused heterocyclic group, 6-membered / 6-membered bicyclic fused heterocyclic group, 5-membered / 6-membered bicyclic fused heterocyclic group or 5-membered / 5-membered bicyclic fused heterocyclic group; Alternatively, ring B is a 5-12 quinary double-ring bridged sub-heterocyclic base; Alternatively, ring B is a 6-9 quinary double-ring bridged subheterocyclic group; Alternatively, ring B is a 7- or 8-membered double-ring bridge sub-heterocyclic base; Alternatively, ring B is selected from The compound of formula (I) according to any one of claims 1-14, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, Ring B is selected from the following structural groups: wherein X1is selected from O, S or CR 6 R 7 ; X2is selected from CR 8 or N; X3is selected from CR 9 or N; R 6 , R 7 , R 8 , R 9 is independently selected from H, deuterium, NH2, OH, halogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, or C3-C6cycloalkyl; s, m, and r are independently selected from 0, 1, or 2; Each R 4 It is independently selected from deuterium, OH, halogen, =O, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy or C3-C6 cycloalkyl; q' is selected from 0 or 1; "q" is selected from 0 or 1. The compound of formula (I) according to any one of claims 1-14, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, Ring B is selected from the following structural groups: wherein Z1is selected from CR 10 or N; Z2is selected from CR 10 or N; R 10 selected from H, deuterium, NH2, OH, halogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, or C3-C6cycloalkyl; t, f, g, and k are independently selected from 0, 1, or 2; Each R 4 It is independently selected from deuterium, OH, halogen, =O, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy or C3-C6 cycloalkyl; q' is selected from 0 or 1; "q" is selected from 0 or 1. The compound of formula (I) according to any one of claims 1-16, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, Ring A is selected from C3-C 10 cycloalkyl, 4-9 membered heterocyclyl, 5-6 membered heteroaryl, or phenyl; Alternatively, ring A can be a 5-10 membered heteroaryl or phenyl group; Alternatively, ring A can be a 5-10 member heteroaryl group; Alternatively, ring A can be a 5-6 membered heteroaryl group; Alternatively, ring A may be selected from phenyl, pyridinyl, pyrimidinyl, or pyridazinyl; Alternatively, ring A is selected from The compound of Formula (I) or a stereoisomer or pharmaceutically acceptable salt thereof according to any one of claims 1-17, wherein, R 1 selected from CN, halogen, Ci-C6alkyl, Ci-C6haloalkyl, or Ci-C6alkoxy, each instance of said Ci-C6alkyl or Ci-C6alkoxy being independently optionally substituted with NH2, OH, halogen, or Ci-C6alkyl; or R is selected from halogen, C1-C6alkyl, C1-C6haloalkyl, or C1-C6alkoxy, each independently optionally substituted with NH2, OH, halogen, or C1-C6alkyl; 1 selected from halogen, C1-C6alkyl, C1-C6haloalkyl, or C1-C6alkoxy, each independently optionally substituted with NH2, OH, halogen, or C1-C6alkyl; or R 1 selected from halogen, Ci-C6alkyl or Ci-C6haloalkyl; Or, R 1 Selected from C1-C6 haloalkyl groups; or R 1 is selected from CN, F, CI, CH3, CHF2, CF3or -OCHF2. The compound of formula (I) according to any one of claims 1-18, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R 4 selected from deuterium, OH, halogen, =0, Ci-C6alkyl or Ci-C6haloalkyl; or R 4 selected from deuterium, =0, or Ci-C6alkyl; or R 4 is selected from deuterium, =0 or CH3. The compound of formula (I) according to any one of claims 1-19, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, p is selected from 0 or 1; Alternatively, p is 1; Alternatively, q can be selected from 0 or 1; Alternatively, q is 0. The compound of Formula (I) or a stereoisomer or pharmaceutically acceptable salt thereof according to any one of claims 1-20, wherein, n is 1 or 2; or, n is 1. The compound of Formula (I) according to claim 1, or a stereoisomer or a pharmaceutically acceptable salt thereof, wherein, The compound of Formula (I) or isomers thereof or pharmaceutically acceptable salts thereof is selected from the following compounds or isomers thereof or pharmaceutically acceptable salts thereof: A pharmaceutical composition comprising a compound of formula (I) as described in any one of claims 1-22, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. Use of a compound of formula (I) as described in any one of claims 1-22, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of claim 23, for the manufacture of a medicament for the prevention or treatment of a GCase-mediated disease. A method of preventing or treating a GCase-mediated disease in a mammal, comprising administering to a mammal, preferably a human, in need of such treatment, a therapeutically effective amount of a compound of formula (I) as described in any one of claims 1-22, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition of claim 23.

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