β-glucocerebrosidase allosteric modulator, pharmaceutical composition thereof and use thereof

Abstract

Provided are compounds represented by formula (I) or stereoisomers or pharmaceutically acceptable salts thereof, a pharmaceutical composition containing same, and the use thereof as a β-glucocerebrosidase (GCase) allosteric modulator, preferably for treating neurodegenerative diseases.

Description

Beta-glucocerebrosidase allosteric modulators, pharmaceutical compositions thereof, and uses thereof

[0001] Cross-reference to Related Applications

[0002] This disclosure claims the benefit of and priority to the following four Chinese patent applications, the entire contents of which are hereby incorporated by reference in their entirety:

[0003] Chinese Patent Application No. CN202410597300.6, filed May 14, 2024, with the China National Intellectual Property Administration,

[0004] Chinese Patent Application No. CN202411292033.8, filed September 14, 2024, with the China National Intellectual Property Administration,

[0005] Chinese Patent Application No. CN202510024333.6, filed January 07, 2025, with the China National Intellectual Property Administration, and

[0006] Chinese Patent Application No. CN202510285716.9, filed March 11, 2025, with the China National Intellectual Property Administration. TECHNICAL FIELD

[0007] The present disclosure belongs to the technical field of medicine, in particular to compounds as allosteric modulators of beta-glucocerebrosidase (also known as acid beta-glucosidase, D-glucosyl-N-acylsphingosine glucosylhydrolase or GCase), pharmaceutical compositions containing the compounds, and uses thereof in the prevention or treatment of neurodegenerative diseases. BACKGROUND

[0008] Beta-glucocerebrosidase is an enzyme with glucosylceramidase activity that metabolizes the glycosphingolipid glucosylceramide (GlcCer) to ceramide and glucose. Beta-glucocerebrosidase has maximum activity at pH 5.5, the pH of the lysosomal compartment. Within the lysosome, it remains membrane-associated and degrades its substrate, glucocerebroside (GluCer). It requires activation by the protein Saposin C and negatively charged lipids to exert maximal catalytic activity, ensuring proper cellular protein degradation and cell signaling.

[0009] Mutations in GBA1 (gene encoding lysosomal enzyme glucocerebrosidase) are one of the most common known genetic risk factors for the development of Parkinson's disease and related synucleinopathies, and are causal for the rare autosomal recessive storage disorder, Gaucher disease. GBA1 mutations can lead to disruption of protein degradation, lysosomal targeting and reduced enzyme performance in lysosomes. Gaucher disease has phenotypic diversity, with both neuronopathic and non-neuronopathic forms. Patients and heterozygous carriers of Gaucher disease have an increased risk of developing Parkinson's disease and dementia with Lewy bodies. Reductions in GCase enzyme activity levels have been detected in different brain regions in Parkinson's disease patients, and furthermore, studies have shown that modulators of GCase enzyme can restore lysosomal function, reduce lipid substrate accumulation, and thus inhibit pathological alpha-Syn aggregation, so GCase modulators are considered as a new drug development strategy for alleviating and treating Parkinson's disease.

[0010] Therefore, it is of great significance to study new, potent and highly selective GCase modulators for the treatment of neurodegenerative diseases. SUMMARY

[0011] The present disclosure provides a compound represented by formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof,

[0012] wherein, is selected from a single bond or a double bond;

[0013] Q is selected from N or C;

[0014] G is selected from N, CH or C(O);

[0015] ring B is selected from the following groups:

[0016] a) C6-C 14 spiro cycloalkylene, C6-C 14 fused cycloalkylene, C5-C 12 bridge cycloalkylene, 6-14 membered spiro heterocyclylene, 6-14 membered fused heterocyclylene or 5-12 membered bridge heterocyclylene;

[0017] or,

[0018] b)

[0019] W is selected from a bond, -CR a R b -, -CR a R b O-, C(O), O or S;

[0020] R a , R b are independently selected from H, halogen, C1-C6alkyl, C1-C6haloalkyl or C1-C4alkoxy;

[0021] Ring A is selected from C3-C 12 cycloalkyl, 4-12 membered heterocyclyl, 5-10 membered heteroaryl, or phenyl;

[0022] each R 1 is independently selected from CN, NH2, OH, halogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, C3-C6cycloalkyl, or 4-10 membered heterocyclyl, said C1-C6alkyl, C1-C6alkoxy, C3-C6cycloalkyl, or 4-10 membered heterocyclyl optionally substituted with NH2, OH, halogen, C1-C6alkyl;

[0023] R 2 is selected from H, C1-C6alkyl, C3-C6cycloalkyl, or 4-10 membered heterocyclyl, said C1-C6alkyl, C3-C6cycloalkyl, or 4-10 membered heterocyclyl optionally substituted with NH2, OH, halogen, C1-C6alkyl, C3-C6cycloalkyl, or 4-6 membered heterocyclyl;

[0024] R 3 is selected from H, C1-C6alkyl, C1-C6alkoxy, C3-C6cycloalkyl, or 4-6 membered heterocyclyl, said C1-C6alkyl, C3-C6cycloalkyl, or 4-6 membered heterocyclyl optionally substituted with NH2, OH, halogen, or C1-C6alkyl;

[0025] each R 4 is independently selected from deuterium, OH, halogen, =O, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, or C3-C6cycloalkyl;

[0026] R 5 is selected from H, deuterium, NH2, CN, OH, halogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, C3-C6cycloalkyl, or 4-7 membered heterocyclyl;

[0027] p, q are independently selected from 0, 1, or 2.

[0028] In some embodiments, in formula (I),

[0029] is selected from a single or double bond;

[0030] Q is selected from N or C;

[0031] G is selected from N, CH, or C(O);

[0032] Ring B is selected from the following groups:

[0033] a) C6-C 14 spiro cycloalkylene, C6-C14 bridged cycloalkyl, 6-14 membered spiro heterocyclyl, 6-14 membered fused heterocyclyl, or 5-12 membered bridged heterocyclyl; 12 bridged cycloalkyl, 6-14 membered spiro heterocyclyl, 6-14 membered fused heterocyclyl, or 5-12 membered bridged heterocyclyl;

[0034] or,

[0035] b)

[0036] W is selected from a bond, -CR a R b -, -CR a R b O- or C(O);

[0037] R a , R b are independently selected from H, halogen, C1-C6alkyl, C1-C6haloalkyl, or C1-C4alkoxy;

[0038] Ring A is selected from C3-C 12 cycloalkyl, 4-12 membered heterocyclyl, or 5-10 membered heteroaryl;

[0039] each R 1 is independently selected from CN, NH2, OH, halogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, C3-C6cycloalkyl, or 4-10 membered heterocyclyl, said C3-C6cycloalkyl or 4-10 membered heterocyclyl optionally substituted with NH2, OH, halogen, C1-C6alkyl;

[0040] R 2 is selected from H, C1-C6alkyl, C3-C6cycloalkyl, or 4-10 membered heterocyclyl, said C1-C6alkyl, C3-C6cycloalkyl, or 4-10 membered heterocyclyl optionally substituted with NH2, OH, halogen, or C1-C6alkyl;

[0041] R 3 is selected from H, C1-C6alkyl, C1-C6alkoxy, C3-C6cycloalkyl, or 4-6 membered heterocyclyl, said C1-C6alkyl, C3-C6cycloalkyl, or 4-6 membered heterocyclyl optionally substituted with NH2, OH, halogen, or C1-C6alkyl;

[0042] each R 4 is independently selected from deuterium, OH, halogen, =O, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, or C3-C6cycloalkyl;

[0043] R 5H, deuterium, NH2, CN, OH, halogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, C3-C6cycloalkyl, or 4-7 membered heterocyclyl;

[0044] p, q are independently selected from 0, 1 or 2.

[0045] In some embodiments, in formula (I),

[0046] is selected from a single bond or a double bond;

[0047] Q is selected from N or C;

[0048] G is selected from N, CH or C(O);

[0049] Ring B is selected from the following groups:

[0050] a) C6-C 14 spiro cycloalkylene, C6-C 14 fused cycloalkylene, C5-C 12 bridged cycloalkylene, 6-14 membered spiro heterocyclylene, 6-14 membered fused heterocyclylene, or 5-12 membered bridged heterocyclylene;

[0051] or,

[0052] b)

[0053] W is selected from a bond, -CR a R b - or -CR a R b O-;

[0054] R a , R b are independently selected from H, halogen, C1-C6alkyl, C1-C6haloalkyl, or C1-C4alkoxy;

[0055] Ring A is selected from C3-C 12 cycloalkyl, or 4-12 membered heterocyclyl;

[0056] each R 1 is independently selected from CN, NH2, OH, halogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, C3-C6cycloalkyl, or 4-10 membered heterocyclyl, said C3-C6cycloalkyl or 4-10 membered heterocyclyl being optionally substituted with NH2, OH, halogen, C1-C6alkyl;

[0057] R 2H, C1-C6alkyl, C3-C6cycloalkyl, or 4-10 membered heterocyclyl, said C1-C6alkyl, C3-C6cycloalkyl, or 4-10 membered heterocyclyl optionally substituted with NH2, OH, halogen, or C1-C6alkyl;

[0058] R 3 H, C1-C6alkyl, C1-C6alkoxy, C3-C6cycloalkyl, or 4-6 membered heterocyclyl, said C1-C6alkyl, C3-C6cycloalkyl, or 4-6 membered heterocyclyl optionally substituted with NH2, OH, halogen, or C1-C6alkyl;

[0059] each R 4 is independently selected from deuterium, OH, halogen, =O, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, or C3-C6cycloalkyl;

[0060] R 5 is selected from H, deuterium, NH2, CN, OH, halogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, C3-C6cycloalkyl, or 4-7 membered heterocyclyl;

[0061] p, q are independently selected from 0, 1, or 2.

[0062] In some embodiments, when is a double bond, Q is C, G is N, and W is selected from a bond or -CR a R b , ring B is not the following structure: and ring A is selected from C3-C 12 cycloalkyl, or 4-12 membered heterocyclyl.

[0063] In some embodiments, when is a double bond, Q is C, G is N, and W is selected from a bond or -CR a R b , ring B is not the following structure:

[0064] In some embodiments, when W is a bond, ring B is not the following structure:

[0065] In some embodiments, when W is selected from -CR a R b , ring B is not the following structure:

[0066] In some embodiments, when W is selected from a bond or -CR a R b , ring B is not the following structure:

[0067] In some embodiments, when W is selected from a bond or -CR a R b then ring B is not the following structure:

[0068] In some embodiments, is a single bond, Q is N, and G is C(O).

[0069] In some embodiments, is a double bond, Q is C, and G is N.

[0070] In some embodiments, ring B is selected from the following groups:

[0071] a) a 6-14 membered spiro- heterocyclylene, 6-14 membered fused- heterocyclylene, or 5-12 membered bridged- heterocyclylene; or,

[0072] b)

[0073] In some embodiments, ring B is selected from the following groups:

[0074] a) a 6-14 membered spiro- heterocyclylene or 6-14 membered fused- heterocyclylene; or,

[0075] b)

[0076] In some embodiments, ring B is selected from the following groups:

[0077] a) a 6-10 membered spiro- heterocyclylene or 6-10 membered fused- heterocyclylene; or,

[0078] b)

[0079] In some embodiments, ring B is selected from a 6-14 membered spiro- heterocyclylene, 6-14 membered fused- heterocyclylene, or 5-12 membered bridged- heterocyclylene.

[0080] In some embodiments, ring B is a 6-14 membered spiro- heterocyclylene or 6-14 membered fused- heterocyclylene.

[0081] In some embodiments, ring B is a 6-14 membered spiro- heterocyclylene or 6-14 membered fused- heterocyclylene each having at least one N atom as a ring-forming atom.

[0082] In some embodiments, ring B is a 6-14 membered N- spiro- heterocyclylene or 6-14 membered N- fused- heterocyclylene.

[0083] In some embodiments, ring B is a 6-14 membered N- spiro- heterocyclylalkyl or 6-14 membered N- fused- heterocyclylalkyl.

[0084] In some embodiments, Ring B is a 6-10 membered N-spiro heterocycloalkyl or a 6-10 membered N-fused heterocycloalkyl.

[0085] In some embodiments, Ring B is selected from a 6-14 membered bicyclic spiro heterocyclyl.

[0086] In some embodiments, Ring B is selected from a 6-10 membered bicyclic spiro heterocyclyl.

[0087] In some embodiments, Ring B is selected from a 3 / 6 bicyclic spiro heterocyclyl, a 4 / 4 bicyclic spiro heterocyclyl, a 4 / 5 bicyclic spiro heterocyclyl, a 4 / 6 bicyclic spiro heterocyclyl, a 5 / 5 bicyclic spiro heterocyclyl, or a 5 / 6 bicyclic spiro heterocyclyl.

[0088] In some embodiments, Ring B is selected from a 5 / 5 bicyclic spiro heterocyclyl, a 4 / 4 bicyclic spiro heterocyclyl, a 4 / 5 bicyclic spiro heterocyclyl, a 5 / 6 bicyclic spiro heterocyclyl, or a 4 / 6 bicyclic spiro heterocyclyl.

[0089] In some embodiments, Ring B is selected from a 4 / 4 bicyclic spiro heterocyclyl, a 4 / 5 bicyclic spiro heterocyclyl, a 5 / 6 bicyclic spiro heterocyclyl, or a 4 / 6 bicyclic spiro heterocyclyl.

[0090] In some embodiments, Ring B is selected from a 4 / 4 bicyclic spiro heterocyclyl, a 4 / 5 bicyclic spiro heterocyclyl, or a 4 / 6 bicyclic spiro heterocyclyl.

[0091] In some embodiments, Ring B is selected from a 6-14 membered bicyclic fused heterocyclyl.

[0092] In some embodiments, Ring B is selected from a 3 / 5 bicyclic fused heterocyclyl, a 6 / 6 bicyclic fused heterocyclyl, a 5 / 6 bicyclic fused heterocyclyl, or a 5 / 5 bicyclic fused heterocyclyl.

[0093] In some embodiments, Ring B is selected from a 6 / 6 bicyclic fused heterocyclyl, a 5 / 6 bicyclic fused heterocyclyl, or a 5 / 5 bicyclic fused heterocyclyl.

[0094] In some embodiments, Ring B is selected from a 3 / 5 bicyclic fused heterocyclyl, a 5 / 6 bicyclic fused heterocyclyl, or a 5 / 5 bicyclic fused heterocyclyl.

[0095] In some embodiments, Ring B is selected from a 5 / 6 bicyclic fused heterocyclyl or a 5 / 5 bicyclic fused heterocyclyl.

[0096] In some embodiments, Ring B is selected from a 5 / 6 bicyclic fused heterocyclyl.

[0097] In some embodiments, Ring B is a 5-12 membered bicyclic bridged heterocyclyl.

[0098] In some embodiments, Ring B is a 6-9 membered bicyclic bridged heterocyclyl. In some embodiments, Ring B is a 7 membered or 8 membered bicyclic bridged heterocyclyl.

[0099] In some embodiments, the 6-14 membered spiro heterocyclyl, 6-14 membered fused heterocyclyl, or 5-12 membered bridged heterocyclyl each independently contains 1, 2, or 3 heteroatoms independently selected from N, O, or S.

[0100] In some embodiments, the 6-14 membered spiro heterocyclyl, 6-14 membered fused heterocyclyl, or 5-12 membered bridged heterocyclyl each independently contains 1, 2, or 3 heteroatoms independently selected from N or O.

[0101] In some embodiments, the 6-14 membered spiro heterocyclyl, 6-14 membered fused heterocyclyl, or 5-12 membered bridged heterocyclyl each independently contains 1, 2, or 3 heteroatoms independently selected from N or O.

[0102] In some embodiments, the 3 / 6 bicyclic spiro heterocyclyl, 4 / 4 bicyclic spiro heterocyclyl, 4 / 5 bicyclic spiro heterocyclyl, 4 / 6 bicyclic spiro heterocyclyl, 5 / 5 bicyclic spiro heterocyclyl, 5 / 6 bicyclic spiro heterocyclyl, 3 / 5 bicyclic fused heterocyclyl, 6 / 6 bicyclic fused heterocyclyl, 5 / 6 bicyclic fused heterocyclyl, 5 / 5 bicyclic fused heterocyclyl, 6-9 bicyclic bridged heterocyclyl, or 7 membered or 8 membered bicyclic bridged heterocyclyl each independently contains 1, 2, or 3 heteroatoms independently selected from N or O.

[0103] In some embodiments, Ring B is selected from C6-C 14 spiro cycloalkyl.

[0104] In some embodiments, Ring B is selected from C6-C 10 spiro cycloalkyl.

[0105] In some embodiments, Ring B is selected from C7-C9 spiro cycloalkyl.

[0106] In some embodiments, Ring B is selected from

[0107] In some embodiments, Ring B is selected from:

[0108] In some embodiments, Ring B is selected from:

[0109] In some embodiments, Ring B is selected from:

[0110] In some embodiments, Ring B is selected from:

[0111] In some embodiments, Ring B is selected from

[0112] In some embodiments, Ring B is selected from

[0113] In some embodiments, is selected from the following structures:

[0114] In some embodiments, is selected from the following structures:

[0115] In some embodiments, is selected from the following structures:

[0116] In some embodiments, is selected from the following structures:

[0117] In some embodiments, is selected from the following structures:

[0118] In some embodiments, is selected from the following structures:

[0119] In some embodiments, W is selected from a bond, -CR a R b -, -CR a R b O-, or C(O).

[0120] In some embodiments, W is selected from a bond, -CR a R b -, -CR a R b O-, C(O), or O.

[0121] In some embodiments, W is selected from a bond, -CH2-, -CHCH3-, -CH2O-, C(O), or O.

[0122] In some embodiments, W is selected from a bond, -CH2-, -CH2O-, or C(O). a R b In some embodiments, W is selected from a bond, -CH2-, -CH2O-, or C(O). a R b O-.

[0123] In some embodiments, W is selected from a bond, -CH2-, -CH2O-, or C(O). a R b O-.

[0124] In some embodiments, W is selected from a bond, -CH2-, -CH2O-, or C(O).

[0125] In some embodiments, W is selected from a bond, -CH2-, -CH2O-, or C(O).

[0126] In some embodiments, W is selected from a bond, -CH2-, -CH2O-, or C(O).

[0127] In some embodiments, W is selected from a bond, -CH2-, -CH2O-, or C(O). a R b In some embodiments, W is selected from a bond, -CH2-, -CH2O-, or C(O). a R b O-.

[0128] In some embodiments, W is selected from a bond, -CH2-, -CH2O-, or C(O).

[0129] In some embodiments, W is C(O).

[0130] In some embodiments, R a , R b are each independently selected from H or C1-C6 alkyl.

[0131] In some embodiments, R a , R b are each H.

[0132] In some embodiments, ring A is selected from C3-C 12 cycloalkyl, 4-12 membered heterocyclyl, or 5-10 membered heteroaryl.

[0133] In some embodiments, ring A is selected from C3-C 12 cycloalkyl, 5-10 membered heteroaryl, or phenyl.

[0134] In some embodiments, ring A is selected from C3-C 12 cycloalkyl, or 5-10 membered heteroaryl.

[0135] In some embodiments, ring A is selected from C5-C10 Bridged cycloalkyl or 5-6 membered heteroaryl.

[0136] In some implementations, ring A is selected from C3-C 12 Cycloalkyl or 4-12 membered heterocyclic groups.

[0137] In some embodiments, ring A is selected from C3-C6 cycloalkyl, 4-9 membered heterocyclic, 5-6 membered heteroaryl, or phenyl.

[0138] In some embodiments, ring A is selected from C3-C6 cycloalkyl, 4-9 membered heterocyclic or 5-6 membered heteroaryl.

[0139] In some implementations, ring A is selected from C5-C. 10 Bridged cycloalkyl, C5-C 10 Spirocycloalkyl or 4-9 membered heterocyclic groups.

[0140] In some implementations, ring A is selected from C3-C 12 Cycloalkyl.

[0141] In some implementations, ring A is selected from C5-C. 10 Bridged cycloalkyl groups.

[0142] In some embodiments, ring A is selected from C5-C6 bridged cycloalkyl groups.

[0143] In some implementations, ring A is

[0144] In some implementations, ring A is selected from 5-7 membered heterocyclic groups.

[0145] In some implementations, ring A is selected from 5-7 quinary bicyclic bridged heterocyclic groups or 5-7 quinary monocyclic heterocyclic groups.

[0146] In some implementations, ring A is selected from 5-7 quinary bicyclic bridged heterocyclic bases.

[0147] In some implementations, ring A is selected from a 6-membered heterocyclic group.

[0148] In some implementations, ring A is selected from a 6-membered monocyclic heterocyclic group.

[0149] In some implementations, ring A is selected from...

[0150] In some implementations, ring A is a 5-10 membered heteroaryl group.

[0151] In some implementations, ring A is a 5-6 membered heteroaryl group.

[0152] In some embodiments, ring A is pyridinyl, thiazolyl, pyrazolyl, or pyrimidinyl.

[0153] In some embodiments, ring A is pyridyl, thiazolyl, or pyrazolyl.

[0154] In some embodiments, ring A is pyridyl.

[0155] In some embodiments, ring A is

[0156] In some embodiments, ring A is

[0157] In some embodiments, ring A is phenyl.

[0158] In some embodiments, ring A is selected from

[0159] In some embodiments, ring A is selected from

[0160] In some embodiments, ring A is selected from

[0161] In some embodiments, ring A is selected from

[0162] In some embodiments, ring A is selected from

[0163] In some embodiments, ring A is selected from

[0164] In some embodiments, ring A is selected from

[0165] In some embodiments, ring A is selected from

[0166] In some embodiments, ring A is selected from

[0167] In some embodiments, each R 1 is independently selected from CN, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, or 4-10 membered heterocyclyl, said C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, or 4-10 membered heterocyclyl optionally substituted with NH2, OH, halogen, C1-C6 alkyl.

[0168] In some embodiments, each R 1independently selected from CN, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, or 4-10 membered heterocyclyl, said C1-C6 alkyl or C1-C6 alkoxy optionally substituted with OH or halogen.

[0169] In some embodiments, each R 1 independently selected from CN, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, or 4-10 membered heterocyclyl, said C1-C6 alkyl or C1-C6 alkoxy optionally substituted with OH or halogen.

[0170] In some embodiments, each R 1 independently selected from halogen, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 alkoxy, said C1-C6 alkoxy optionally substituted with halogen.

[0171] In some embodiments, each R 1 independently selected from C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 alkoxy, said C1-C6 alkoxy optionally substituted with halogen.

[0172] In some embodiments, each R 1 independently selected from CN, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, or 4-6 membered heterocyclyl, said C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, or 4-6 membered heterocyclyl optionally substituted with OH or halogen.

[0173] In some embodiments, each R 1 independently selected from CN, NH2, OH, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C3-C6 cycloalkyl, or 4-10 membered heterocyclyl, said C3-C6 cycloalkyl or 4-10 membered heterocyclyl optionally substituted with NH2, OH, halogen, or C1-C6 alkyl.

[0174] In some embodiments, R 1 is selected from halogen, C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 alkoxy, said C1-C6 alkyl, C1-C6 alkoxy optionally substituted with NH2, OH, halogen, or C1-C6 alkyl.

[0175] In some embodiments, R 1 is selected from C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 alkoxy.

[0176] In some embodiments, R1 halogen, C1-C6 alkyl or C1-C6 haloalkyl.

[0177] In some embodiments, R 1 is C1-C6 alkyl.

[0178] In some embodiments, R 1 is C1-C6 haloalkyl.

[0179] In some embodiments, R 1 is selected from F, CH3, CF3, CH2CF3, OCHF2, CN, cyclopropyl or morpholinyl.

[0180] In some embodiments, R 1 is selected from F, CH3, CF3, CH2CF3, or OCHF2.

[0181] In some embodiments, R 1 is selected from CH3, CF3, or OCHF2.

[0182] In some embodiments, R 1 is selected from F, CH3, or CF3.

[0183] In some embodiments, R 1 is CH3.

[0184] In some embodiments, R 1 is CF3.

[0185] In some embodiments, is selected from the following structures:

[0186] In some embodiments, is selected from the following structures:

[0187] In some embodiments, is selected from the following structures:

[0188] In some embodiments, is selected from the following structures:

[0189] In some embodiments, is selected from the following structures:

[0190] In some embodiments, is selected from the group consisting of:

[0191] In some embodiments, is selected from the group consisting of:

[0192] In some embodiments, is selected from the group consisting of:

[0193] In some embodiments, R 2 is selected from H, C1-C6 alkyl, C3-C6 cycloalkyl, or 4-10 membered heterocyclyl, optionally substituted with NH2, OH, halogen, or C1-C6 alkyl.

[0194] In some embodiments, R 2 is selected from C1-C6 alkyl, C3-C6 cycloalkyl, or 4-10 membered heterocyclyl, optionally substituted with NH2, OH, halogen, C1-C6 alkyl, C3-C6 cycloalkyl, or 4-6 membered heterocyclyl.

[0195] In some embodiments, R 2 is selected from C1-C6 alkyl, C3-C6 cycloalkyl, or 4-10 membered heterocyclyl, optionally substituted with NH2, OH, halogen, or C1-C6 alkyl.

[0196] In some embodiments, R 2 is selected from C1-C6 alkyl, or 4-10 membered heterocyclyl, optionally substituted with halogen.

[0197] In some embodiments, R 2 is selected from C1-C6 alkyl, C3-C6 cycloalkyl, or 4-6 membered heterocyclyl, optionally substituted with NH2, OH, halogen, C1-C6 alkyl, C3-C6 cycloalkyl, or 4-6 membered heterocyclyl.

[0198] In some embodiments, R 2 is selected from C1-C6 alkyl, C3-C6 cycloalkyl, or 4-6 membered heterocyclyl, optionally substituted with NH2, OH, halogen, or C1-C6 alkyl.

[0199] In some embodiments, R2 C1-C6alkyl substituted with halogen.

[0200] In some embodiments, R 2 C1-C6alkyl substituted with F.

[0201] In some embodiments, R 2 is selected from CF3, CHF2, CH2CHF2, cyclobutyl, or oxetanyl. In some embodiments, R 2 is selected from CH2CHF2, cyclobutyl, oxetanyl, or -CH2-oxetanyl.

[0202] In some embodiments, R 2 is selected from CH2CHF2or oxetanyl.

[0203] In some embodiments, R 2 is selected from CH2CHF2, cyclobutyl, or oxetanyl.

[0204] In some embodiments, R 2 is CH2CHF2.

[0205] In some embodiments, R 3 is selected from H, C1-C6alkyl, or C1-C6alkoxy.

[0206] In some embodiments, R 3 is H.

[0207] In some embodiments, R 4 is selected from deuterium, OH, halogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, or C3-C6cycloalkyl.

[0208] In some embodiments, R 4 is selected from deuterium, OH, halogen, =O, C1-C6alkyl, or C1-C6haloalkyl.

[0209] In some embodiments, R 4 is selected from deuterium, =O, or C1-C6alkyl.

[0210] In some embodiments, R 4 is selected from =O or C1-C6alkyl.

[0211] In some embodiments, R 4 is selected from deuterium, =O, or CH3.

[0212] In some embodiments, R 4 is selected from =O or CH3.

[0213] In some embodiments, R 4selected from deuterium or =0.

[0214] In some embodiments, R 4 is =0.

[0215] In some embodiments, R 5 is selected from H, deuterium, halogen, C1-C6alkyl, or C3-C6cycloalkyl.

[0216] In some embodiments, R 5 is selected from H, halogen, C1-C6alkyl, or C3-C6cycloalkyl.

[0217] In some embodiments, R 5 is selected from H, deuterium, halogen, or C3-C6cycloalkyl.

[0218] In some embodiments, R 5 is selected from H or C1-C6alkyl.

[0219] In some embodiments, R 5 is selected from H, F, Cl, CH3, or cyclopropyl.

[0220] In some embodiments, R 5 is selected from H, F, Cl, or cyclopropyl.

[0221] In some embodiments, R 5 is selected from H, Cl, CH3, or cyclopropyl.

[0222] In some embodiments, R 5 is selected from H or CH3.

[0223] In some embodiments, R 5 is H.

[0224] In some embodiments, p is selected from 0 or 1.

[0225] In some embodiments, q is selected from 0 or 1.

[0226] In some embodiments, the compound of Formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, is selected from a compound of Formula (I-1) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0227] Ring A is selected from C3-C 12 cycloalkyl, or 4-12 membered heterocyclyl;

[0228] when W is selected from a bond or -CR a R b , then Ring B is not the following structure:

[0229] Ring B, R 1 , R 2 , R 3 , R 4 , R 5 , p, q, W are defined as above.

[0230] In some embodiments of formula (I-1), ring A is selected from C3-C 12 Cycloalkyl or 5-10 membered heterocyclyl.

[0231] In some embodiments of formula (I-1), ring A is selected from C5-C7 cycloalkyl or 5-7 membered heterocyclyl.

[0232] In some embodiments, the compound of formula (I) or formula (I-1), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, is selected from a compound represented by formula (II-1), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0233] X1is selected from O, S, or CR 6 R 7 ;

[0234] X2is selected from CR 8 or N;

[0235] X3is selected from C, CR 9 or N;

[0236] R 6 , R 7 , R 8 , R 9 are independently selected from H, deuterium, NH2, OH, halogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, or C3-C6cycloalkyl;

[0237] s, m, n are independently selected from 0, 1, or 2;

[0238] q' is selected from 0 or 1;

[0239] q" is selected from 0 or 1;

[0240] ring A is selected from C3-C 12 cycloalkyl or 4-12 membered heterocyclyl;

[0241] when W is selected from a bond or -CR a R b -, ring B is not the following structure:

[0242] R 1 , R 2 , R 3 , R 4R 5 p, q, W are as defined above.

[0243] In some embodiments of formula (II-1), R 6 R 7 R 8 R 9 is independently selected from H or C1-C6 alkyl.

[0244] In some embodiments, the compound of formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, is selected from a compound of formula (V-1) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0245] Ring A is selected from C3-C6 cycloalkyl, 4-9 membered heterocyclyl, or 5-6 membered heteroaryl;

[0246] R 1 R 2 R 3 R 4 R 5 p, q, W are as defined above.

[0247] In some embodiments, the compound of formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, is selected from a compound of formula (I-2) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0248] Ring A, Ring B, R 1 R 2 R 3 R 4 R 5 p, q, W are as defined above.

[0249] In some embodiments of formula (I-2), Ring B is selected from C6-C 14 spiro cycloalkylene, 6-14 membered spiro heterocyclylene, or 6-14 membered fused heterocyclylene;

[0250] W is selected from a bond, -CR a R b -, -CR a R b O-, C(O), or O;

[0251] R a R b is independently selected from H or C1-C6 alkyl.

[0252] Ring A is selected from C3-C 12 cycloalkyl, 4-12 membered heterocyclyl, 5-10 membered heteroaryl, or phenyl;

[0253] each R 1 is independently selected from CN, halogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, C3-C6cycloalkyl, or 4-10 membered heterocyclyl, said C1-C6alkyl and C1-C6alkoxy optionally substituted with OH or halogen;

[0254] R 2 is selected from C1-C6alkyl or 4-10 membered heterocyclyl, said C1-C6alkyl optionally substituted with halogen;

[0255] R 3 is selected from H, C1-C6alkyl or C1-C6alkoxy;

[0256] R 4 is selected from =O or C1-C6alkyl;

[0257] R 5 is selected from H or C1-C6alkyl.

[0258] In some embodiments of formula (I-2), ring B is selected from 6-14 membered spiro- heterocyclylene or 6-14 membered fused-heterocyclylene;

[0259] In some embodiments, the compound of formula (I) or formula (I-2), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, is selected from a compound represented by formula (II-2), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0260] X1is selected from O, S, or CR 6 R 7 ;

[0261] X2is selected from CR 8 or N;

[0262] X3is selected from C, CR 9 or N;

[0263] 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;

[0264] s, m, n are independently selected from 0, 1, or 2;

[0265] q' is selected from 0 or 1;

[0266] q" is selected from 0 or 1;

[0267] ring A, R1 , R 2 , R 3 , R 4 , R 5 , p, W are defined as described above.

[0268] In some embodiments, X1is selected from O or CR 6 R 7 .

[0269] In some embodiments, X2is selected from CH or N.

[0270] In some embodiments, is a single bond, X3is selected from CR 9 or N.

[0271] In some embodiments, at least one of X2and X3is N.

[0272] In some embodiments, R 6 , R 7 , R 8 , R 9 are independently selected from H, deuterium, or C1-C6alkyl.

[0273] In some embodiments, R 6 , R 7 , R 8 , R 9 are all independently H.

[0274] In some embodiments, s, m, n are independently selected from 0 or 1.

[0275] In some embodiments, q’ is 0; q” is 0.

[0276] In some embodiments, is a single bond.

[0277] In some embodiments, the compound of Formula (I) or Formula (I-2), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, is selected from a compound of Formula (III-2), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0278] T is selected from N or CH;

[0279] Ring A, R 1 , R 2 , R 3 , R 4 , R 5 , p, q, W are defined as described above.

[0280] In some embodiments, T is N.

[0281] In some embodiments, the compound of Formula (I) or Formula (I-2), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, is selected from a compound of Formula (V-2), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0282] Ring A, R 1 , R 2 , R 3 , R 4 , R 5 , p, q, W are defined as above.

[0283] In some embodiments, the compound of Formula (I) or Formula (I-2), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, is selected from a compound of Formula (VI-2), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0284] Z1is selected from CR 10 or N;

[0285] Z2is selected from CR 10 or N;

[0286] R 10 is selected from H, deuterium, NH2, OH, halogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, or C3-C6cycloalkyl;

[0287] t, f, g, k are independently selected from 0, 1, or 2;

[0288] q' is selected from 0 or 1;

[0289] q" is selected from 0 or 1;

[0290] Ring A, R 1 , R 2 , R 3 , R 4 , R 5 , p, W are defined as above.

[0291] In some embodiments, R 10 is selected from H, deuterium, or C1-C6alkyl.

[0292] In some embodiments, R 10 is H.

[0293] In some embodiments, Z1is N, and Z2is selected from CH or N.

[0294] In some embodiments, Z1is CH or N, and Z2is N.

[0295] In some embodiments, Z1is N, Z2is CH. In some embodiments, Z1is CH, Z2is selected from N. In some embodiments, Z1is N, Z2is N.

[0296] In some embodiments, at least one of Z1and Z2is N.

[0297] In some embodiments, t is 1.

[0298] In some embodiments, k is 1.

[0299] In some embodiments, f, g are selected from 0 or 1.

[0300] In some embodiments, f, g are both 0.

[0301] In some embodiments, f, g are both 1.

[0302] In some embodiments, q' is 0, q" is 0.

[0303] In some embodiments, the compound of Formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof is selected from a compound of Formula (II) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0304] X1is selected from O, S, or CR 6 R 7 ;

[0305] X2is selected from CR 8 or N;

[0306] X3is selected from C, CR 9 or N;

[0307] R 6 , R 7 , R 8 , R 9 are independently selected from H, deuterium, NH2, OH, halogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, or C3-C6cycloalkyl;

[0308] s, m, n are independently selected from 0, 1, or 2;

[0309] q' is selected from 0 or 1;

[0310] q" is selected from 0 or 1;

[0311] Ring A, R 1 , R 2 , R 3 , R 4 , R 5, p, W, Q, G are defined as above.

[0312] In some embodiments, the compound of Formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof is selected from a compound of Formula (III) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0313] T is selected from N or CH;

[0314] Ring A, R 1 , R 2 , R 3 , R 4 , R 5 , p, q, W, Q, G are defined as above.

[0315] In some embodiments, the compound of Formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof is selected from a compound of Formula (IV) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0316] Y1, Y2are independently selected from N or CH;

[0317] Z is selected from CH2or CH2CH2;

[0318] Ring A, R 1 , R 2 , R 3 , R 4 , R 5 , p, q, W, Q, G are defined as above.

[0319] In some embodiments, the compound of Formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof is selected from a compound of Formula (V) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0320] Ring A, R 1 , R 2 , R 3 , R 4 , R 5 , p, q, W, Q, G are defined as above.

[0321] In some embodiments, the compound of Formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof is selected from a compound of Formula (VI) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0322] Z1is selected from CR 10or N;

[0323] Z2is selected from CR 10 or N;

[0324] R 10 is selected from H, deuterium, NH2, OH, halogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, or C3-C6cycloalkyl;

[0325] t, f, g, k are independently selected from 0, 1, or 2;

[0326] q' is selected from 0 or 1;

[0327] q" is selected from 0 or 1;

[0328] Ring A, R 1 , R 2 , R 3 , R 4 , R 5 , p, W, Q, G are as defined above.

[0329] In some embodiments, the compound of Formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, is selected from the following compounds or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0330] In some embodiments, the compound of Formula (I) or Formula (I-2) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, is selected from the following compounds or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0331] In another aspect, the present disclosure provides a pharmaceutical composition comprising a compound of Formula (I), Formula (I-1), Formula (II-1), Formula (V-1), Formula (I-2), Formula (II-2), Formula (III-2), Formula (V-2), Formula (VI-2), Formula (II), Formula (III), Formula (IV), Formula (V), or Formula (VI) of the present disclosure, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0332] In another aspect, the present disclosure provides a method of preventing or treating a disease mediated by GCase in a subject (e.g., a mammal), comprising administering to a subject, preferably a mammal, more preferably a human, in need of such treatment, a therapeutically effective amount of a compound of Formula (I), Formula (I-1), Formula (II-1), Formula (V-1), Formula (I-2), Formula (II-2), Formula (III-2), Formula (V-2), Formula (VI-2), Formula (II), Formula (III), Formula (IV), Formula (V), or Formula (VI), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0333] In another aspect, the present disclosure provides a method of preventing or treating a disease mediated by GCase in a subject (e.g., a mammal), comprising administering to a subject, preferably a mammal, more preferably a human, in need of such treatment, a therapeutically effective amount of a compound of Formula (I), Formula (I-1), Formula (II-1), Formula (V-1), Formula (I-2), Formula (II-2), Formula (III-2), Formula (V-2), Formula (VI-2), Formula (II), Formula (III), Formula (IV), Formula (V), or Formula (VI), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0334] In another aspect, the present disclosure provides a use of a compound of Formula (I), Formula (I-1), Formula (II-1), Formula (V-1), Formula (I-2), Formula (II-2), Formula (III-2), Formula (V-2), Formula (VI-2), Formula (II), Formula (III), Formula (IV), Formula (V), or Formula (VI), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in the preparation of a medicament for preventing or treating a disease mediated by GCase.

[0335] In another aspect, the present disclosure provides a use of a compound of Formula (I), Formula (I-1), Formula (II-1), Formula (V-1), Formula (I-2), Formula (II-2), Formula (III-2), Formula (V-2), Formula (VI-2), Formula (II), Formula (III), Formula (IV), Formula (V), or Formula (VI), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in the preparation of a medicament for preventing or treating a disease mediated by GCase.

[0336] In another aspect, the present disclosure provides a use of a compound of Formula (I), Formula (I-1), Formula (II-1), Formula (V-1), Formula (I-2), Formula (II-2), Formula (III-2), Formula (V-2), Formula (VI-2), Formula (II), Formula (III), Formula (IV), Formula (V), or Formula (VI), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in the prevention or treatment of a disease mediated by GCase.

[0337] In yet another aspect, the present disclosure provides a use of a compound of Formula (I), Formula (I-1), Formula (II-1), Formula (V-1), Formula (I-2), Formula (II-2), Formula (III-2), Formula (V-2), Formula (VI-2), Formula (II), Formula (III), Formula (IV), Formula (V), or Formula (VI), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in allosterically modulating GCase.

[0338] In another aspect, the present disclosure provides a compound of Formula (I), Formula (I-1), Formula (II-1), Formula (V-1), Formula (I-2), Formula (II-2), Formula (III-2), Formula (V-2), Formula (VI-2), Formula (II), Formula (III), Formula (IV), Formula (V), or Formula (VI), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for use in preventing or treating a GCase-mediated disease.

[0339] In yet another aspect, the present disclosure provides a compound of Formula (I), Formula (I-1), Formula (II-1), Formula (V-1), Formula (I-2), Formula (II-2), Formula (III-2), Formula (V-2), Formula (VI-2), Formula (II), Formula (III), Formula (IV), Formula (V), or Formula (VI), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for use in allosterically modulating GCase.

[0340] In some embodiments, the GCase-mediated disease is a disease that benefits from allosteric modulation of GCase. In some embodiments, the GCase-mediated disease is a disease that benefits from activation of GCase.

[0341] In some embodiments, the GCase-mediated disease is selected from neurodegenerative diseases.

[0342] In some embodiments, the neurodegenerative disease is Alzheimer’s disease or Parkinson’s disease.

[0343] Any embodiment of any aspect of the present disclosure can be combined with any other embodiment of the same aspect or other aspects, without appearing contradictory. In addition, any technical feature in any embodiment of any aspect of the present disclosure can be applied to the same technical feature in other embodiments, without appearing contradictory.

[0344] Definitions and explanations

[0345] Unless otherwise indicated, the terms used in the present disclosure have the following meanings, the definitions of the groups and terms recited in the present disclosure, including the definitions as examples, exemplary definitions, preferred definitions, definitions recited in the tables, definitions of specific compounds in the examples, etc., can be combined and incorporated with each other arbitrarily. A particular term should not be considered indefinite or unclear without a specific definition, but should be understood according to the ordinary meaning in the art. When a trade name appears herein, it is intended to refer to its corresponding product or active ingredient thereof.

[0346] Herein represents a point of attachment.

[0347] The term "stereoisomer" refers to isomers that have the same structure except for the arrangement of atoms in space, including cis-trans isomers, enantiomers, and diastereomers.

[0348] The compounds of the present disclosure can have asymmetric atoms such as carbon atoms, sulfur atoms, nitrogen atoms, phosphorus atoms, or asymmetric double bonds, and thus the compounds of the present disclosure can exist in specific geometric or stereoisomeric forms. The specific geometric or stereoisomeric forms can be cis and trans isomers, E and Z geometric isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, as well as racemic mixtures or other mixtures thereof, for example, enantiomeric or diastereomeric enrich mixtures, all of which are within the scope of the compounds of the present disclosure. Additional asymmetric carbon atoms, asymmetric sulfur atoms, asymmetric nitrogen atoms, or asymmetric phosphorus atoms can be present in a substituent group, and all such isomers and mixtures thereof are included within the scope of the compounds of the present disclosure. The compounds of the present disclosure containing asymmetric atoms can be isolated in optically active form or as racemic mixtures, and the optically active forms can be obtained either by separation of racemic mixtures or by synthesis using chiral reagents or chiral starting materials.

[0349] The term "substituted" means that any one or more hydrogen atoms on a particular atom is replaced with a substituent group, provided that the valence of the particular atom is normal and the resulting compound is stable. When the substituent is oxo (i.e., =O), it means that two hydrogen atoms are replaced, and oxo does not occur on an aromatic group.

[0350] The term "optionally" or "optional" means that the subsequently described event or circumstance can or can not occur, and this description includes instances where the event or circumstance occurs and instances where it does not. For example, ethyl "optionally" substituted with halogen means that the ethyl group can be unsubstituted (CH2CH3), mono-substituted (CH2CH2F, CH2CH2C1, etc.), multi-substituted (CHFCH2F, CH2CHF2, CHFCH2C1, CH2CHC12, etc.), or fully substituted (CF2CF3, CF2CC13, CC12CC13, etc.). It will be understood by those skilled in the art that, for any group containing one or more substituents, such group does not include a substitution or substitution pattern that is not possible

[0351] When any variable (e.g., R a , R b ) occurs more than one time in a compound, its definition in each instance is independent of its definition at every other place in which it occurs. b b For example, if a group is substituted with 2 R a , each R b is selected independently.

[0352] When a substituent is attached to a ring atom via a bond that is crossed into the plane of the ring, the substituent can be bonded to any atom of the ring.

[0353] When a linking group referred to herein is not indicated to be attached in a particular orientation, it can be attached in either orientation. For example, when W in structural unit ring A-W-ring B is selected from "-CR a R b O-" then W can be attached to form either the unit "ring A-CR a R b O-ring B" or the unit "ring A-OCR m R n -ring B".

[0354] Herein, a bond drawn as a solid line and a dashed line represents a single or double bond. For example, the structural unit contains

[0355] Herein, "bond" in "W is selected from a bond" means a direct bond.

[0356] Herein, C m -C n means an integer number of carbon atoms in the range of m-n. For example, "C1-C 10" is intended to mean that the group can have 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, 8 carbon atoms, 9 carbon atoms, or 10 carbon atoms.

[0357] The term "alkyl" refers to a hydrocarbon group of formula C n H 2n+1 The term "C1-C6alkyl" can be understood to mean an alkyl group having 1, 2, 3, 4, 5, or 6 carbon atoms, specific examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, neopentyl, hexyl, 2-methylpentyl, and the like. The term "C1-C3alkyl" can be understood to mean a straight or branched chain saturated alkyl group having 1, 2, or 3 carbon atoms. The "C1-C3alkyl" group can include the "C1-C6alkyl" group, which can further include the "C1-C3alkyl" group. 10 The term "C1-C6alkyl" can be understood to mean an alkyl group having 1, 2, 3, 4, 5, or 6 carbon atoms, specific examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, neopentyl, hexyl, 2-methylpentyl, and the like. The term "C1-C3alkyl" can be understood to mean a straight or branched chain saturated alkyl group having 1, 2, or 3 carbon atoms. The "C1-C 10 The term "C1-C6alkyl" can be understood to mean an alkyl group having 1, 2, 3, 4, 5, or 6 carbon atoms, specific examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, neopentyl, hexyl, 2-methylpentyl, and the like. The term "C1-C3alkyl" can be understood to mean a straight or branched chain saturated alkyl group having 1, 2, or 3 carbon atoms. The "C1-C

[0358] The term "haloalkyl" is intended to include mono-haloalkyl and poly-haloalkyl groups. For example, the term "C1-C6haloalkyl" means a C1-C6alkyl group as defined above substituted by one or more halogen atoms, including but not limited to trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, trichloromethyl, pentafluoroethyl, penta chloroethyl, and the like.

[0359] The term "alkoxy" refers to a straight or branched chain alcohol in which the hydrogen atom on the hydroxyl group is replaced by an alkyl group, and can be understood as "alkyloxy" or "alkyl-O-". The term "C1-C6alkoxy" can be understood as "C1-C6alkyloxy" or "C1-C6alkyl-O-". The "C1-C6alkoxy" group can include the "C1-C3alkyl" group. 10 The term "C1-C6alkoxy" can be understood as "C1-C6alkyloxy" or "C1-C6alkyl-O-". The "C1-C6alkoxy" group can include the "C1-C3alkyl" group. 10 The term "C1-C6alkoxy" can be understood as "C1-C6alkyloxy" or "C1-C6alkyl-O-". The "C1-C6alkoxy" group can include the "C1-C3alkyl" group. 10 The term "C1-C6alkoxy" can be understood as "C1-C6alkyloxy" or "C1-C6alkyl-O-". The "C1-C6alkoxy" group can include the "C1-C3alkyl" group.10 Alkoxy" can include "Ci-C6alkoxy" and "Ci-C3alkoxy" and the like ranges, which "Ci-C6alkoxy" can further include "Ci-C3alkoxy".

[0360] The term "cycloalkyl" refers to a carbocyclic ring that is fully saturated and exists as a monocyclic, fused, bridged, or spirocyclic, etc. Unless otherwise indicated, the carbocyclic ring is typically a 3- to 14-membered ring. The term "C3-C 12 Cycloalkyl" can be understood to mean 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 said 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, and the like. The term "C3-C 12 Cycloalkyl" can include "C3-C6cycloalkyl", the term "C3-C6cycloalkyl" can be understood to mean 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, and the like.

[0361] The term "spirocycloalkyl" refers to a 6- to 14-membered polycyclic group sharing one carbon atom (termed the spiro atom) between single rings, which can contain one or more double bonds, but does not have a fully conjugated pi-electron system overall. Preferred are 7- to 12- (e.g., 7-, 8-, 9-, 10-, 11-, or 12-membered). More preferred are 4-membered / 4-membered, 4-membered / 5-membered, 4-membered / 6-membered, 5-membered / 5-membered, or 5-membered / 6-membered bicyclic spirocycloalkyl groups, where A-membered / B-membered bicyclic spirocycloalkyl refers to a spirocyclic ring formed by an A-membered monocyclic ring and a B-membered monocyclic ring sharing one carbon atom (termed the spiro atom). Specific examples of spirocycloalkyl groups include, but are not limited to:

[0362] The term "spirocycloalkyl" refers to a 6- to 14-membered polycyclic group sharing one carbon atom (termed the spiro atom) between single rings, which can contain one or more double bonds, but does not have a fully conjugated pi-electron system overall. Preferred are 7- to 12- (e.g., 7-, 8-, 9-, 10-, 11-, or 12-membered). More preferred are 4-membered / 4-membered, 4-membered / 5-membered, 4-membered / 6-membered, 5-membered / 5-membered, or 5-membered / 6-membered bicyclic spirocycloalkyl groups, where A-membered / B-membered bicyclic spirocycloalkyl refers to a spirocyclic ring formed by an A-membered monocyclic ring and a B-membered monocyclic ring sharing one carbon atom (termed the spiro atom). Specific examples of spirocycloalkyl groups include, but are not limited to:

[0363] The term "fused cycloalkyl" refers to a 6- to 14-membered polycyclic group in which each ring in the system 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 as a whole does not have a fully conjugated π-electron system. 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, wherein A-membered / B-membered bicyclic fused cycloalkyl refers to a polycyclic group in which an A-membered monocyclic ring and a B-membered monocyclic ring share an adjacent pair of carbon atoms. Specific examples of fused cycloalkyl groups include, but are not limited to:

[0364] The term “condensed cycloalkyl” refers to a divalent group derived from a “condensed cycloalkyl” group as defined herein.

[0365] 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 constituent 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:

[0366] The term “bridged cycloalkyl” refers to a divalent group derived from a “bridged cycloalkyl” group as defined herein.

[0367] The term "heterocyclyl" refers to a monocyclic, fused, spiro, or bridged ring that is fully saturated or partially saturated (not aromatic overall as a heteroaromatic) having 1, 2, 3, 4, or 5 heteroatoms or heteroatom groups (i.e., groups of atoms containing a heteroatom) in its ring atoms, including but not limited to a nitrogen atom (N), an oxygen atom (O), a sulfur atom (S), a phosphorus atom (P), a boron atom (B), -S(=0)2-, -S(=0)-, -P(=0)2-, -P(=0)-, -NH-, -S(=0)(=NH)-, -C(=0)NH-, or -NHC(=0)NH-, etc. The term "4-12 membered heterocyclyl" refers to a heterocyclyl having 4, 5, 6, 7, 8, 9, 10, 11, or 12 ring atoms, and having 1, 2, 3, 4, or 5 ring atoms independently selected from the heteroatoms or heteroatom groups described above. "3-10 membered heterocyclyl" includes "4-7 membered heterocyclyl", wherein specific examples of 4-membered heterocyclyl include but are not limited to azetidinyl, thietanyl, or oxetanyl; specific examples of 5-membered heterocyclyl include but are not limited to tetrahydrofuranyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl, 4,5-dihydrooxazolyl, or 2,5-dihydro-lH-pyrrolyl; specific examples of 6-membered heterocyclyl include but are not limited to tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, trithianyl, tetrahydropyridinyl, or 4H-[l,3,4]thiadiazinyl; specific examples of 7-membered heterocyclyl include but are not limited to diazepanyl. The heterocyclyl can also be a bicyclic group, wherein specific examples of 5,5 membered bicyclic groups include but are not limited to hexahydrocyclopenta[c]pyrrol-2(lH)-yl or ; specific examples of 5,6 membered bicyclic groups include but are not limited to hexahydropyrrolo[l,2-a]pyrazin-2(lH)-yl, 5,6,7,8-tetrahydro-[l,2,4]triazolo[4,3-a]pyrazinyl, or 5,6,7,8-tetrahydroimidazo[l,5-a]pyrazinyl. Optionally, the heterocyclyl can be a benzo-fused ring of the above 4-7 membered heterocyclyl, specific examples include but are not limited to dihydroisoquinolinyl, etc. "4-10 membered heterocyclyl" can include "5-10 membered heterocyclyl", "4-7 membered heterocyclyl", "5-6 membered heterocyclyl", "6-8 membered heterocyclyl", "4-10 membered heterocycloalkyl", "5-10 membered heterocycloalkyl", "4-7 membered heterocycloalkyl", "5-6 membered heterocycloalkyl", "6-8 membered heterocycloalkyl", etc. "4-7 membered heterocyclyl" can further include "4-6 membered heterocyclyl", "5-6 membered heterocyclyl", "4-7 membered heterocycloalkyl", "4-6 membered heterocycloalkyl", "5-6 membered heterocycloalkyl", etc. Although some bicyclic heterocyclyl groups in the present disclosure contain a benzene ring or a heteroaromatic ring partially, the heterocyclyl is still non-aromatic overall.

[0368] The term "spiroheterocyclyl" refers to a polycyclic heterocyclic group of 6 to 14 members sharing one atom (referred to as the spiro atom) between single rings, wherein one or more ring atoms are a heteroatom selected from a nitrogen atom (N), an oxygen atom (O), a sulfur atom (S), S(O), or S(O)2, with the remaining ring atoms being carbon. It can contain one or more double bonds, but as a whole does not have a fully conjugated pi-electron system. Preferably, it is 7 to 12 members (e.g., 7, 8, 9, 10, 11, or 12 members). 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 spiroheterocyclyl. Wherein A-membered / B-membered bicyclic spiroheterocyclyl refers to a spiro ring formed by an A-membered monocyclic ring and a B-membered monocyclic ring sharing one atom (referred to as the spiro atom). Specific examples of spiroheterocyclyl groups include, but are not limited to:

[0369] The term "spiroheterocyclyl" refers to a polycyclic heterocyclic group of 6 to 14 members sharing one atom (referred to as the spiro atom) between single rings, wherein one or more ring atoms are a heteroatom selected from a nitrogen atom (N), an oxygen atom (O), a sulfur atom (S), S(O), or S(O)2, with the remaining ring atoms being carbon. It can contain one or more double bonds, but as a whole does not have a fully conjugated pi-electron system. Preferably, it is 7 to 12 members (e.g., 7, 8, 9, 10, 11, or 12 members). 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 spiroheterocyclyl. Wherein A-membered / B-membered bicyclic spiroheterocyclyl refers to a spiro ring formed by an A-membered monocyclic ring and a B-membered monocyclic ring sharing one atom (referred to as the spiro atom). Specific examples of spiroheterocyclyl groups include, but are not limited to:

[0370] The term "N-spiroheterocyclyl" refers to a spiroheterocyclyl group as defined above comprising a nitrogen atom as the point of attachment.

[0371] The term "fused heterocyclyl" refers to a polycyclic heterocyclic group of 6 to 14 members, each ring in the system sharing an adjacent pair of atoms with other rings in the system, wherein one or more ring atoms are a heteroatom selected from a nitrogen atom (N), an oxygen atom (O), a sulfur atom (S), S(O), or S(O)2, with the remaining ring atoms being carbon. It can contain one or more double bonds, but as a whole does not have a fully conjugated pi-electron system. Preferably, it is 7 to 12 members (e.g., 7, 8, 9, 10, 11, or 12 members). It can be a bicyclic, tricyclic, tetracyclic, or polycyclic fused heterocyclyl, preferably a bicyclic or tricyclic, more preferably a 5-membered / 5-membered or 5-membered / 6-membered bicyclic fused heterocyclyl. Wherein A-membered / B-membered bicyclic fused heterocyclyl refers to a fused ring formed by an A-membered monocyclic ring and a B-membered monocyclic ring sharing an adjacent pair of atoms. Specific examples of fused heterocyclyl groups include, but are not limited to:

[0372] The term "fused heterocyclyl" refers to a polycyclic heterocyclic group of 6 to 14 members, each ring in the system sharing an adjacent pair of atoms with other rings in the system, wherein one or more ring atoms are a heteroatom selected from a nitrogen atom (N), an oxygen atom (O), a sulfur atom (S), S(O), or S(O)2, with the remaining ring atoms being carbon. It can contain one or more double bonds, but as a whole does not have a fully conjugated pi-electron system. Preferably, it is 7 to 12 members (e.g., 7, 8, 9, 10, 11, or 12 members). It can be a bicyclic, tricyclic, tetracyclic, or polycyclic fused heterocyclyl, preferably a bicyclic or tricyclic, more preferably a 5-membered / 5-membered or 5-membered / 6-membered bicyclic fused heterocyclyl. Wherein A-membered / B-membered bicyclic fused heterocyclyl refers to a fused ring formed by an A-membered monocyclic ring and a B-membered monocyclic ring sharing an adjacent pair of atoms. Specific examples of fused heterocyclyl groups include, but are not limited to:

[0373] The term "N-fused heterocyclyl" refers to a fused heterocyclyl group as defined above comprising a nitrogen atom as the point of attachment.

[0374] The term "bridged heterocyclyl" refers to a 5- to 12-membered, polycyclic heterocyclic radical, sharing any two non-adjacent ring atoms of two rings, which can contain one or more double bonds, but which does not have a fully conjugated pi-electron system overall, wherein 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 5- to 9-membered (e.g., 5-, 6-, 7-, 8-, or 9-membered). Depending on the number of rings comprising the ring system, it can be referred to as bicyclic, tricyclic, tetracyclic, or polycyclic bridged heterocyclyl, preferably bicyclic, tricyclic, or tetracyclic, more preferably bicyclic or tricyclic. Specific examples of bridged heterocyclyl groups include, but are not limited to:

[0375] The term "bridged heterocyclyl" refers to a 5- to 12-membered, polycyclic heterocyclic radical, sharing any two non-adjacent ring atoms of two rings, which can contain one or more double bonds, but which does not have a fully conjugated pi-electron system overall, wherein 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 5- to 9-membered (e.g., 5-, 6-, 7-, 8-, or 9-membered). Depending on the number of rings comprising the ring system, it can be referred to as bicyclic, tricyclic, tetracyclic, or polycyclic bridged heterocyclyl, preferably bicyclic, tricyclic, or tetracyclic, more preferably bicyclic or tricyclic. Specific examples of bridged heterocyclyl groups include, but are not limited to:

[0376] The term "aryl" refers to all-carbon monocyclic or fused polycyclic ring systems having a completely conjugated pi-electon system. Aryl groups can have 6-20 carbon atoms, 6-14 carbon atoms, or 6-12 carbon atoms. The term "C6-C 20 aryl" is understood to mean aryl groups having 6 to 20 carbon atoms. In particular, a ring having 6 carbon atoms ("C6aryl"), such as, for example, a phenyl group, or a ring having 9 carbon atoms ("C9aryl"), such as, for example, an indane or indenyl group, or a ring having 10 carbon atoms ("C 10 aryl"), such as, for example, a tetrahydronaphthyl, dihydronaphthyl or naphthyl group, or a ring having 13 carbon atoms ("C 13 aryl"), such as, for example, a fluorene group, or a ring having 14 carbon atoms ("C 14 aryl"), such as, for example, an anthracene group. The term "C6-C 10 aryl" is understood to mean aryl groups having 6 to 10 carbon atoms. In particular, a ring having 6 carbon atoms ("C6aryl"), such as, for example, a phenyl group, or a ring having 9 carbon atoms ("C9aryl"), such as, for example, an indane or indenyl group, or a ring having 10 carbon atoms ("C 10 aryl"), such as, for example, a tetrahydronaphthyl, dihydronaphthyl or naphthyl group. The term "C6-C 20 aryl" can comprise "C6-C 10 aryl".

[0377] The term "heteroaryl" refers to a monocyclic or fused polycyclic ring system having aromaticity, which contains at least one ring atom selected from N, O, S, the remaining ring atoms being C. The term "5-10 membered heteroaryl" is understood to include monocyclic or bicyclic aromatic ring systems having 5, 6, 7, 8, 9 or 10 ring atoms, in particular 5 or 6 or 9 or 10 ring atoms, and which contain 1, 2, 3, 4 or 5, preferably 1, 2 or 3 heteroatoms independently selected from N, O and S. In particular, the heteroaryl group is selected from thienyl, furanyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl or thiadiazolyl and the like as well as their benzo derivatives, such as, for example, benzofuranyl, benzothienyl, benzothiazolyl, benzoxazolyl, benzoisoxazolyl, benzoimidazolyl, benzotriazolyl, indazolyl, indolyl or isoindolyl and the like; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl or triazinyl and the like as well as their benzo derivatives, such as, for example, quinolinyl, quinazolinyl or isoquinolinyl and the like; or azocinyl, indolizinyl, purinyl and the like as well as their benzo derivatives; or cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, pteridinyl, carbazolyl, acridinyl, phenoxazinyl, phenothiazinyl or phenoxazinyl and the like. The term "5-6 membered heteroaryl" refers to an aromatic ring system having 5 or 6 ring atoms, and which contain 1, 2 or 3, preferably 1-2 heteroatoms independently selected from N, O and S.

[0378] The term "halo" or "halogen" refers to fluorine, chlorine, bromine or iodine.

[0379] The term "treatment" means the administration of a compound or formulation described herein to improve or eliminate a disease or one or more symptoms associated with the disease, and includes:

[0380] (i) inhibiting the disease or condition, i.e., arresting its development;

[0381] (ii) relieving the disease or condition, i.e., causing regression of the disease or condition.

[0382] The term "prevention" means the administration of a compound or formulation described herein to prevent a disease or one or more symptoms associated with the disease, including preventing the disease or condition from occurring in a mammal, in particular, when such mammal is predisposed to the condition, but has not yet been diagnosed as having it.

[0383] The term "therapeutically effective amount" means:

[0384] (i) treating a particular disease, condition or disorder, (ii) reducing, ameliorating or eliminating one or more symptoms of a particular disease, condition or disorder, or (iii) delaying the onset of one or more symptoms of a particular disease, condition or disorder described herein, of a compound of the present disclosure.

[0385] The amount of a compound of the present disclosure that will constitute a "therapeutically effective amount" will vary depending on the compound, the condition and its severity, the mode of administration, and the age of the mammal to be treated, but can be determined routinely by one of ordinary skill in the art having regard to his own knowledge and to this disclosure.

[0386] The term "pharmaceutically acceptable" in reference to those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0387] The term "pharmaceutically acceptable salt" refers to pharmaceutically acceptable acid or base addition salts, including salts with inorganic or organic acids, as well as salts with inorganic or organic bases.

[0388] The term "pharmaceutical composition" refers to a mixture of one or more compounds of the present disclosure or salts thereof with a pharmaceutically acceptable excipient. The purpose of a pharmaceutical composition is to facilitate administration of a compound of the present disclosure to an organism.

[0389] The term "pharmaceutically acceptable excipient" refers to those excipients that are not biologically or otherwise undesirable, and that do not interfere with the biological activity of the active compound. Suitable excipients are well known to those skilled in the art, e.g., carbohydrates, waxes, water soluble and / or swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, and the like.

[0390] The term "individual" includes mammals. Examples of mammals include, but are not limited to, any member of the mammalian class: humans, non-human primates (such as chimpanzees and other apes and monkey species); farm animals such as cattle, horses, sheep, goats, swine; domestic animals such as rabbits, dogs, and cats; laboratory animals including rodents, such as rats, mice and guinea pigs, and the like. Examples of non-human mammals include, but are not limited to, birds and fish, and the like. In one embodiment of the methods and compositions provided herein, the mammal is a human.

[0391] The words "comprise" or "comprising" and variations such as "comprises" or "comprising", when used in this document, can be understood to encompass the terms "consisting of" or "consisting of" and variations thereof, such as "consists" or "consisting of".

[0392] The present disclosure also includes isotopically-labeled compounds of the present disclosure which are identical to those recited herein, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into compounds of the present disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorous, sulfur, fluorine, iodine, and chlorine, such as 2 H, 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, and the like.

[0393] Certain isotopically-labeled compounds of the present disclosure (for example, those 3 H and 14 C) can be used in compound and / or substrate tissue distribution analysis. Tritiated (i.e., 3 H) and carbon-14 (i.e., 14 C) isotopes are particularly preferred for 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. Isotopically-labeled compounds of the present disclosure can generally be prepared by

[0394] The pharmaceutical compositions of the present disclosure can be prepared by combining a compound of the present disclosure with a suitable pharmaceutically acceptable excipient, such as can be formulated into solid, semi-solid, liquid, or gaseous dosage forms, such as tablets, pills, capsules, powders, granules, ointments, emulsions, suspensions, suppositories, injections, inhalers, gels, microspheres, aerosols, and the like.

[0395] Typical routes of administering a compound of the present disclosure, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, include, but are not limited to, oral, rectal, topical, inhalation, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, intravenous administration.

[0396] 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.

[0397] 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.

[0398] 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.

[0399] The pharmaceutical composition may also be suitable for parenteral administration, such as in suitable unit dosage forms of sterile solutions, suspensions or lyophilized products.

[0400] In all methods of administration of the compounds of general formula (I) or formula (I-1) or formula (V-1) or formula (I-2) or their stereoisomers or pharmaceutically acceptable salts described herein, the daily dose is from 0.01 mg / kg to 1000 mg / kg body weight, in the form of single or separate doses. Detailed Implementation

[0401] 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.

[0402] Unless otherwise specified, all reagents used in this disclosure are commercially available and can be used without further purification.

[0403] Unless otherwise stated, the proportions expressed for mixed solvents are volume-based.

[0404] Unless otherwise stated, % refers to wt%.

[0405] Compounds are processed manually or Software naming conventions are used; commercially available compounds use supplier catalog names.

[0406] The structure of the compound is determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). The unit of NMR shift is 10 -6 (ppm). The solvent for NMR determination is deuterated dimethyl sulfoxide, deuterated chloroform, deuterated methanol, etc., and the internal standard is tetramethylsilane (TMS). "IC 50 " refers to the half maximal inhibitory concentration, which refers to the concentration at which the effect of the maximum inhibition is halved.

[0407] The eluent below can form a mixed eluent from two or more solvents, and the ratio is the volume ratio of each solvent, such as "0-7% tetrahydrofuran / petroleum ether", which means that during the gradient elution process, the volume ratio of tetrahydrofuran / petroleum ether in the mixed eluent is 0:100-7:93.

[0408] Abbreviations:

[0409] THF: tetrahydrofuran; LiAlH4: lithium aluminum hydride; DCM: dichloromethane; TEA: triethylamine; Ms2O: methanesulfonic anhydride; DMF: N,N- dimethylformamide; TFA: trifluoroacetic acid; Na2CO3: sodium carbonate; MeOH: methanol; NaBH3CN sodium cyanoborohydride; Py: pyridine; EDCI: l-(3- dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride; BOC: tert-butoxycarbonyl; DMSO: dimethylsulfoxide; Pd(dppf)Cl2: [l,l'-bis(diphenylphosphino) ferrocene] palladium dichloride; dioxane: dioxane; ACN: acetonitrile; DBU: 1,8- diazabicyclo[5.4.0]undec-7-ene; EtOH: ethanol; NaIO4: sodium periodate; NaBH3CN: sodium cyanoborohydride; K2OsO4: potassium osmium; EA: ethyl acetate; EPhos Pd G4: (methylsulfonic acid {bicyclohexyl(3-isopropoxy-2',4',6'-triisopropyl-[l,l'- biphenyl]-2-yl)phosphane}(2'-methylamino-l,l'-biphenyl-2-yl)palladium(II); EPhos: bicyclohexyl(3-isopropoxy-2',4',6'-triisopropyl-[l,l'-biphenyl]-2-yl)phosphane; HATU: 2-(7-azabenzotriazol-l-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate; DMA: N,N-dimethyl aniline; Pd2(dba)3: tris(dibenzylideneacetone)dipalladium; Xantphos: 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene; DIPEA or DIEA: N,N-diisopropylethylamine; Rf: ratio of the migration distance of the solute to the migration distance of the mobile phase; LiHDMS: lithium bis(trimethylsilyl)amide; TOSMIC: methyl phenylsulfonyl methyl isocyanide; t-BuOK: potassium tert-butoxide; Gphos G6 Pd: [2-(dicyclohexylphosphino)-3-tert-butoxy-6-methoxy-2',6'- diisopropyl-l,l'-biphenyl](4-((2-(trimethylsilyl)ethoxy)carbonyl)phenyl-l-yl)palladium bromide.

[0410] Example 1: 6-(6-(Bicyclo[l.l.l]pentan-l-ylmethoxy)-2-azaspiro[3.3]heptan-2-yl)-l-(2,2- difluoroethyl)-lH-pyrazolo[3,4-b]pyrazine

[0411] Step 1: Synthesis of Intermediate 1-2

[0412] Compound 1-1 (1.00 g) was dissolved in 15 mL of tetrahydrofuran under nitrogen protection, and a tetrahydroaluminum lithium solution (2.5 M, 5.35 mL) was added dropwise at 0 °C. After stirring at room temperature for 16 hours, the reaction was completed. The reaction solution was quenched by adding water (2 mL), 15% NaOH (2 mL), and H2O (6 mL) at 0 °C, and anhydrous magnesium sulfate was added and stirred for 30 minutes. After filtration, the filtrate was concentrated to obtain intermediate 1-2 (1.03 g).

[0413] 1 H-NMR (400 MHz, DMSO-d6) δ 4.47 (t, J = 5.5 Hz, 1H), 3.36 (d, J = 5.6 Hz, 2H), 2.54 (s, 1H), 1.70 (s, 6H).

[0414] Step 2: Synthesis of intermediate 1-3

[0415] Intermediate 1-2 (0.90 g) was dissolved in 10 mL of dichloromethane under nitrogen protection, and triethylamine (2.78 g) and methanesulfonic anhydride (3.19 g) were added at 0 °C. After stirring at room temperature for 16 hours, the reaction was completed. To the reaction solution was added 20 mL of dichloromethane, which was washed with saturated sodium bicarbonate (10 mL*3), and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain intermediate 1-3 (1.46 g).

[0416] 1 H-NMR (400 MHz, DMSO-d6) δ 4.47 (t, J = 5.5 Hz, 1H), 3.36 (d, J = 5.6 Hz, 2H), 2.54 (s, 1H), 1.70 (s, 6H).

[0417] Step 3: Synthesis of intermediate 1-4

[0418] tert-Butyl 6-hydroxy-2-azaspiro[3.3]heptane-2-carboxylate (1.05 g) was dissolved in N,N-dimethylformamide (15 mL) and cooled to 0 °C, and sodium hydride (295 mg, 60% purity) was slowly added under a nitrogen atmosphere. After stirring for 30 minutes, intermediate 1-3 (1.30 g) was added. After stirring at room temperature for 16 hours, the reaction was completed. 15 mL of saturated ammonium chloride solution was slowly added, followed by extraction with ethyl acetate (20 mL*3), and the combined organic phase was washed with saturated brine (10 mL*2). After drying over anhydrous sodium sulfate, the organic phase was concentrated. The crude product was purified by silica gel column chromatography (gradient 0-7% tetrahydrofuran / petroleum ether) to obtain intermediate 1-4 (0.62 g).

[0419] 1H-NMR (400 MHz, DMSO-d6) δ 3.85-3.70 (m, 5H), 3.18 (s, 2H), 2.48 (s, 1H), 2.39-2.37 (m, 2H), 1.98-1.95 (m, 2H), 1.67 (s, 6H), 1.36 (s, 9H).

[0420] Step 4: synthesis of intermediate 1-5

[0421] Intermediate 1-4 (0.62 g) was dissolved in 8 mL of dichloromethane, trifluoroacetic acid (2.24 g) was added, the reaction was completed after the reaction solution was stirred at room temperature for 2 hours. The reaction solution was concentrated to obtain intermediate 1-5 (0.43 g), which was directly used in the next step reaction.

[0422] Step 5: synthesis of compound 1

[0423] Intermediate 1-5 (196 mg) and 6-chloro-1-(2,2-difluoroethyl)-1H-pyrazolo[3,4-b]pyrazine (100 mg) were dissolved in N,N-dimethylformamide (2 mL), and sodium carbonate (145 mg) was added. The reaction was completed after being stirred at 100°C for 16 hours. 10 mL of water was added, and ethyl acetate (10 mL*2) was extracted. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by preparative high performance liquid chromatography (column: C18 150*40 mm; mobile phase: [acetonitrile-water (0.5‰ ammonia water + 0.01‰ ammonium bicarbonate)]; acetonitrile ratio: 40%-80%) to obtain the title compound 1 (57.2 mg).

[0424] 1 H-NMR (400 MHz, DMSO-d6) δ 8.14 (s, 1H), 7.88 (s, 1H), 6.43 (tt, J = 3.7, 54.9 Hz, 1H), 4.66 (td, J = 15.0, 3.8 Hz, 2H), 4.17 (s, 2H), 4.12 (m, 2H), 3.91-3.84 (m, 1H), 3.22 (s, 2H), 2.55-2.50 (m, 2H), 2.50 (s, 1H), 2.13-2.04 (m, 2H), 1.70 (s, 6H).

[0425] MS m / z (ESI): 376.1.

[0426] Example 2: 6-(2-(Bicyclo[1.1.1]pentan-1-yl)-2,7-diazaspiro[3.5]nonan-7-yl)-1-(2,2- difluoroethyl)-1H-pyrazolo[3,4-b]pyrazine

[0427] Step 1: Synthesis of intermediate 2-2

[0428] Compound 2-1 (500 mg) and bicyclo[l.l.l]-l-pentanamine hydrochloride (228 mg) were added to 5 mL of methanol under nitrogen protection, stirred at 60 °C for 16 hours, then sodium cyanoborohydride (240 mg) was added, the reaction was completed after continuing to stir at 60 °C for 16 hours. The reaction was quenched by adding 10 mL of saturated aqueous sodium bicarbonate solution and 1 mL of 15% aqueous sodium hydroxide solution, extracted with ethyl acetate (10 mL*3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography (fast silica gel column, gradient 0-80% petroleum ether / tetrahydrofuran) to obtain intermediate 2-2 (220 mg). 12g .

[0429] MS m / z (ESI): 237.2 [M-56] +

[0430] Step 2: Synthesis of intermediate 2-3

[0431] Intermediate 2-2 (220 mg) and 1 mL of trifluoroacetic acid were added to 3 mL of dichloromethane under nitrogen protection, and the reaction was completed after stirring at 25 °C for 12 hours. The reaction was concentrated to obtain the crude product of intermediate 2-3 (250 mg), which was directly used in the next reaction.

[0432] MS m / z (ESI): 193.3 [M+H] + .

[0433] Step 3: Synthesis of compound 2

[0434] Intermediate 2-3 (150 mg) and 6-chloro-l-(2,2-difluoroethyl)-lH-pyrazolo[3,4-b]pyrazine (221 mg) were dissolved in N,N-dimethylformamide (2 mL), and sodium carbonate (413 mg) was added. The reaction was completed after stirring at 100 °C for 12 hours. 10 mL of water was added, and the mixture was extracted with ethyl acetate (10 mL*2). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by preparative high performance liquid chromatography (Phenomenex Gemini NX 150*30 mm, 5 μm; mobile phase: [acetonitrile-water (2.25% formic acid)]; acetonitrile ratio: 27%-67%) to obtain the title compound 2 (25.5 mg).

[0435] MS m / z (ESI): 375.2 [M+H] + .

[0436] 1H NMR (400 MHz, DMSO-d6) δ 8.45 (s, 1H), 8.11 (s, 1H), 6.43 (tt, J = 54.9, 3.8 Hz, 1H), 4.68 (td, J = 15.0, 3.8 Hz, 2H), 3.77 - 3.61 (m, 4H), 2.98 (s, 4H), 2.39 (s, 1H), 1.75 - 1.69 (m, 4H), 1.66 (s, 6H).

[0437] Example 3: 6-(2-(Bicyclo[l. l. l ]pentan- l -ylmethyl)-2,7-diazaspiro[3.5]nonan-7-yl)- l -(2,2- difluoroethyl)- lH-pyrazolo[3,4-b]pyrazine

[0438] Step 1: Synthesis of intermediate 3-2

[0439] Compound 3-1 (1.00 g), 2,7-diazaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester (2.02 g) were dissolved in 30 mL of pyridine, l-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (3.42 g) was added at 0 °C, the reaction was stirred at 25 °C for 16 hours. The reaction was concentrated to get the crude product, which was purified by silica gel column chromatography (gradient 0-12% tetrahydrofuran / petroleum ether) to give intermediate 3-2 (2.40 g).

[0440] MS m / z (ESI): 321.1 [M+H] +

[0441] 1 H NMR (400 MHz, DMSO-d6) δ 3.93 (s, 2H), 3.55 (s, 2H), 3.30 - 3.22 (m, 4H), 2.42 (s, 1H), 2.00 (s, 6H), 1.64 - 1.58 (m, 4H), 1.39 (s, 9H).

[0442] Step 2: Synthesis of intermediate 3-3

[0443] Intermediate 3-2 (370 mg) was dissolved in 5 mL of tetrahydrofuran, under nitrogen atmosphere, borane / tetrahydrofuran solution (1 M, 3.46 mL) was added slowly dropwise. After the addition was completed, the reaction was warmed to 65 °C and stirred for 16 hours. LCMS showed the reaction was complete. The reaction was quenched by slowly adding 10 mL of methanol, the reaction was concentrated to get the crude product (350 mg), which was used directly in the next step.

[0444] Step 3: Synthesis of intermediate 3-4

[0445] Intermediate 3-3 (350 mg) was dissolved in 6 mL of dichloromethane, trifluoroacetic acid (2.30 g) was added, and the reaction was completed after stirring at 25 °C for 16 hours. The reaction solution was concentrated to obtain the target crude product (285 mg), which was directly used in the next step.

[0446] MS m / z (ESI): 207.1 [M+H] +

[0447] Step 4: Synthesis of compound 3

[0448] Intermediate 3-4 (94.4 mg) and 6-chloro-1-(2,2-difluoroethyl)-1H-pyrazolo[3,4- b]pyrazine (100 mg) were dissolved in N,N-dimethylformamide (2 mL), and sodium carbonate (146 mg) was added. The reaction was completed after stirring at 100 °C for 16 hours. 10 mL of water was added, and extraction was performed with ethyl acetate (10 mL*2). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by preparative high-performance liquid chromatography (C18 150*40 mm; mobile phase: [acetonitrile-water (0.5‰ ammonia water + 0.01‰ ammonium bicarbonate)]; acetonitrile ratio: 12%-52%) to obtain the title compound 3 (65.5 mg).

[0449] MS m / z (ESI): 389.1 [M+H] +

[0450] 1 H NMR (400 MHz, DMSO-d6) δ 8.45 (s, 1H), 8.12 (s, 1H), 6.43 (tt, J = 54.9, 3.8 Hz, 1H), 4.68 (td, J = 15.0, 3.8 Hz, 2H), 3.79-3.63 (m, 4H), 2.98 (s, 4H), 2.45 (s, 1H), 2.42 (s, 2H), 1.79-1.70 (m, 4H), 1.66 (s, 6H).

[0451] Example 4: 1-(2,2-difluoroethyl)-6-(2-(1-methyl-2-oxabicyclo[2.1.1]hexan-4-yl)-2,7- diazaspiro[3.5]nonan-7-yl)-1H-pyrazolo[3,4-b]pyrazine

[0452] According to the synthetic method of Reference Example 2, the bicyclo[1.1.1]-1-pentanamine hydrochloride in Step 1 was replaced by (1-methyl-2-oxabicyclo[2.1.1]hexan-4-amine hydrochloride), and the title compound 4 was prepared in the same manner.

[0453] MS m / z (ESI): 405.2 [M+H] + .

[0454] 1 H NMR (400 MHz, DMSO-d6) δ 8.46 (s, 1H), 8.12 (s, 1H), 6.43 (tt, J = 54.9, 3.8 Hz, 1H), 4.69 (td, J = 15.0, 3.8 Hz, 2H), 3.80 - 3.67 (m, 4H), 3.40 (s, 2H), 3.25 (br s, 4H), 1.83 - 1.69 (m, 6H), 1.50 - 1.49 (m, 2H), 1.36 (s, 3H).

[0455] Example 5: 7-(Bicyclo[l. l. l ]pentan- l -ylmethoxy)-2-( l -(2,2-difluoroethyl)- lH- pyrazolo[3,4-b]pyrazin-6-yl)-5-oxa-2-azaspiro[3.4]octane

[0456] Reference Example 1, Step 3, tert-butyl 6-hydroxy-2-azaspiro[3.3]heptane-2- carboxylate was replaced by tert-butyl 6-hydroxy-2-azaspiro[3.4]octane-2-carboxylate. (tert-butyl 7-hydroxy-5-oxa-2-azaspiro[3.4]octane-2-carboxylate) was prepared in the same manner as Reference Example 1, Step 4 to give Compound 5.

[0457] 1 H NMR (400 MHz, DMSO-d6) δ 8.46 (s, 1H), 8.12 (s, 1H), 6.43 (tt, J = 54.9, 3.8 Hz, 1H), 4.69 (td, J = 15.0, 3.8 Hz, 2H), 3.80 - 3.67 (m, 4H), 3.40 (s, 2H), 3.25 (br s, 4H), 1.83 - 1.69 (m, 6H), 1.50 - 1.49 (m, 2H), 1.36 (s, 3H).

[0458] MS m / z (ESI): 405.2 [M+H]

[0459] Example 6: 6-(2-(Bicyclo[l. l. l ]pentan- l -ylmethoxy)-7-azaspiro[3.5]nonan-7-yl)- l -(2,2- difluoroethyl)- lH-pyrazolo[3,4-b]pyrazine

[0460] Reference Example 1, Step 3, tert-butyl 6-hydroxy-2-azaspiro[3.3]heptane-2- carboxylate was replaced by tert-butyl 6-hydroxy-2-azaspiro[3.4]octane-2-carboxylate. (2-hydroxy-7-azaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester), compound 6 was prepared in the same manner.

[0461] 1 H NMR (400 MHz, DMSO-d6) δ 8.45 (s, 1H), 8.11 (s, 1H), 6.43 (tt, J = 54.9, 3.8 Hz, 1H), 4.68 (td, J = 15.0, 3.8 Hz, 2H), 4.11 - 3.88 (m, 1H), 3.78 - 3.57 (m, 4H), 3.21 (s, 2H), 2.49 (s, 1H), 2.26 - 2.13 (m, 2H), 1.70 (s, 6H), 1.69 - 1.54 (m, 6H).

[0462] MS m / z (ESI): 404.3.

[0463] Example 7: 6-(2-(Bicyclo[1.1.1]pentan-1-ylmethoxy)-6-azaspiro[3.4]octan-6-yl)-1-(2,2- difluoroethyl)-1H-pyrazolo[3,4-b]pyrazine

[0464] Reference Example 1 was replaced by (2-hydroxy-6-azaspiro[3.4]octane-6-carboxylic acid tert-butyl ester), compound 7 was prepared in the same manner.

[0465] 1 H NMR (400 MHz, DMSO-d6) δ 8.45 (s, 1H), 8.11 (s, 1H), 6.43 (tt, J = 54.9, 3.8 Hz, 1H), 4.68 (td, J = 15.0, 3.8 Hz, 2H), 4.11 - 3.88 (m, 1H), 3.78 - 3.57 (m, 4H), 3.21 (s, 2H), 2.49 (s, 1H), 2.26 - 2.13 (m, 2H), 1.70 (s, 6H), 1.69 - 1.54 (m, 6H).

[0466] MS m / z (ESI): 404.3.

[0467] Example 8: 6-(2-(Bicyclo[1.1.1]pentan-1-ylmethyl)-2-azaspiro[3.5]non-6-en-7-yl)-1-(2,2- difluoroethyl)-1H-pyrazolo[3,4-b]pyrazine

[0468] Step 1: Synthesis of Intermediate 8-2

[0469] Compound 8-1 (300 mg) and bis(pinacolato)diboron (697 mg) were dissolved in 5 mL of dioxane, potassium acetate (404 mg) and [1,1'-bis(diphenylphosphino)ferrocene] palladium dichloride (167 mg) were added. The reaction was stirred at 80 °C for 2 hours under nitrogen protection. The reaction was complete. The reaction was concentrated and purified by silica gel column chromatography (petroleum ether / ethyl acetate gradient elution) to give the title compound 8-2 (425 mg).

[0470] MS m / z (ESI): 229.1 [M+H-82] +

[0471] Step 2: synthesis of intermediate 8-3

[0472] Intermediate 8-2 (425 mg) and 7-((trifluoromethyl)sulfonyl)oxy)-2- azaspiro[3.5]non-6-ene-2-carboxylic acid tert-butyl ester (300 mg) were dissolved in 5 mL of dioxane and 1 mL of water, cesium carbonate (668 mg) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride dichloromethane complex (111 mg) were added. The reaction was stirred at 90 °C for 2 hours under nitrogen protection. The reaction was complete. 15 mL of water was added, extracted with ethyl acetate (15 mL*3), dried over anhydrous sodium sulfate and concentrated under reduced pressure. The crude product was purified by flash silica gel column chromatography (gradient 0-13% petroleum ether / tetrahydrofuran, flow rate 30 mL / min) to give intermediate 8-3 (270 mg). 4g flash silica gel column, gradient 0-13% petroleum ether / tetrahydrofuran, flow rate 30 mL / min), to give intermediate 8-3 (270 mg).

[0473] MS m / z (ESI): 406.3 [M+H] +

[0474] Step 3: synthesis of intermediate 8-4

[0475] Intermediate 8-3 (218 mg) was dissolved in 3 mL of dichloromethane, trifluoroacetic acid (1.23 g) was added. The reaction was stirred at 30 °C for 2 hours. The reaction was complete by LC-MS monitoring. The reaction was concentrated under reduced pressure to give the target product crude intermediate 8-4 (164 mg).

[0476] MS m / z (ESI): 306.1 [M+H] +

[0477] Step 4: synthesis of compound 8

[0478] The crude intermediate 8-4 (165 mg) prepared in the previous step and bicyclo[l.l.l]pentane-l-ylmethanesulfonic acid methyl ester (123 mg) were dissolved in 3 mL of acetonitrile, and potassium carbonate (224 mg) was added. The reaction was complete after the reaction solution was heated to 80 °C and stirred for 16 hours. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative high-performance liquid chromatography (Welch Xtimate C18 150*25 mm*5 um; mobile phase: [acetonitrile-water (0.5‰ ammonia water + 0.01‰ ammonium bicarbonate)]; acetonitrile ratio: 60%-80%) to obtain compound 8 (10.6 mg).

[0479] 1 H NMR (400 MHz, DMSO-d6) δ 8.97 (s, 1H), 8.48 (s, 1H), 7.21-6.81 (m, 1H), 6.51 (tt, J = 54.7, 3.6 Hz, 1H), 4.91 (td, J = 15.2, 3.7 Hz, 2H), 3.10-2.89 (m, 4H), 2.72-2.58 (m, 2H), 2.43 (s, 1H), 2.41 (s, 2H), 1.88 (t, J = 6.3 Hz, 2H), 1.64 (s, 6H).

[0480] MS m / z (ESI): 386.3 [M+H] +

[0481] Example 9: 6-(4-(Bicyclo[l.l.l]pentan-l-ylmethyl)piperazin-l-yl)-l-(2,2- difluoroethyl)-lH-pyrazolo[3,4-b]pyrazine

[0482] The title compound 9 was prepared by the same method as in Reference Example 3, using 2,7-diazaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester in Step 1 instead of (tert-butyl piperazine-1-carboxylate).

[0483] 1 H NMR (400 MHz, DMSO-d6) δ 8.44 (s, 1H), 8.14 (s, 1H), 6.44 (tt, J = 54.9, 3.9 Hz, 1H), 4.69 (td, J = 15.0, 3.9 Hz, 2H), 3.73 (t, J = 5.1 Hz, 4H), 2.55-2.51 (m, 4H), 2.49 (s, 1H), 2.38 (s, 2H), 1.76 (s, 6H).

[0484] MS m / z (ESI): 349.1 [M+H] +

[0485] Example 10: 6-(2-(Bicyclo[1.1.1]pentan-1-ylmethyl)-2-azaspiro[3.5]nonan-7-yl)-1-(2,2- difluoroethyl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0486] Step 1: Synthesis of intermediate 10-2

[0487] Compound 10-1 (2.15 g) and 7-amino-2-azaspiro[3.5]nonane-2-carboxylic acid tert-butyl ester (1.1 g) were dissolved in 50 mL of tetrahydrofuran, and 1,8-diazabicyclo[5.4.0]undec-7-ene (1.39 g) was added at 0 °C. The reaction was complete after stirring at 60 °C for 16 hours. The reaction solution was concentrated under reduced pressure. The crude product was purified by flash column chromatography on silica gel (20 g) using a gradient of 0-14% petroleum ether / tetrahydrofuran at a flow rate of 100 mL / min to give intermediate 10-2 (1.9 g). 20g flash column chromatography on silica gel (20 g) using a gradient of 0-14% petroleum ether / tetrahydrofuran at a flow rate of 100 mL / min to give intermediate 10-2 (1.9 g).

[0488] 1 H NMR (400 MHz, DMSO-d6) δ 9.00 (d, J = 7.7 Hz, 1H), 8.38 (s, 1H), 6.62-6.15 (m, 1H), 4.80-4.60 (m, 2H), 3.85-3.70 (m, 1H), 3.61-3.49 (m, 4H), 1.90-1.69 (m, 4H), 1.61-1.47 (m, 2H), 1.37 (s, 9H), 1.34-1.21 (m, 2H).

[0489] MS m / z (ESI): 388.0 [M+H-56] +

[0490] Step 2: Synthesis of intermediate 10-3

[0491] Intermediate 10-2 (1.9 g) was dissolved in a mixture of 32 mL of ethanol and 8 mL of water, and iron powder (1.20 g) and ammonium chloride (2.29 g) were added. The reaction was complete after stirring at 80 °C for 1 hour. The reaction solution was filtered and concentrated under reduced pressure. 20 mL of water was added, and the solution was extracted with ethyl acetate (20 mL*3). After drying over anhydrous sodium sulfate, the solution was filtered and concentrated under reduced pressure to give crude intermediate 10-3 (1.6 g).

[0492] MS m / z (ESI): 358.4 [M+H-56] +

[0493] Step 3: Synthesis of intermediate 10-4

[0494] The crude intermediate 10-3 (700 mg) prepared in the previous step was dissolved in 3 mL of formamide. The reaction was stirred at 120 °C for 16 hours until completion. The reaction was concentrated under reduced pressure to give the crude product, which was purified by flash silica gel column chromatography (gradient 0-25% petroleum ether / tetrahydrofuran, flow rate 80 mL / min) to give intermediate 10-4 (270 mg). 40g Flash silica gel column, gradient 0-25% petroleum ether / tetrahydrofuran, flow rate 80 mL / min) to give intermediate 10-4 (270 mg).

[0495] MS m / z (ESI): 368.3 [M+H-56] +

[0496] Step 4: Synthesis of intermediate 10-5

[0497] Intermediate 10-4 (210 mg) was dissolved in 3.0 mL of dichloromethane, then trifluoroacetic acid (1.13 g) was added. The reaction was stirred at 30 °C for 2 hours until completion by LC-MS monitoring. The reaction was concentrated under reduced pressure to give intermediate 10-5 (160 mg).

[0498] MS m / z (ESI): 324.2 [M+H] +

[0499] Step 5: Synthesis of compound 10

[0500] Intermediate 10-5 (130 mg) and bicyclo[1.1.1]pentan-1-yl methanesulfonate (92.1 mg) were dissolved in 2 mL of acetonitrile, then potassium carbonate (166 mg) was added. The reaction was stirred at 80 °C for 16 hours until completion. The reaction was filtered and concentrated to give the crude product, which was purified by preparative high-performance liquid chromatography (column: Welch Xtimate C18 150*25mm*5um; mobile phase: [acetonitrile-water (0.01‰ ammonium bicarbonate)]; acetonitrile ratio: 50%-70%) to give compound 10 (24.5 mg).

[0501] 1H NMR (400 MHz, DMSO-d6) δ 8.31 (s, 1H), 8.13 (s, 1H), 6.43 (tt, J = 54.9, 3.8 Hz, 1H), 5.03 (td, J = 14.6, 3.8 Hz, 2H), 4.74 - 4.47 (m, 1H), 3.02 (s, 2H), 2.87 (s, 2H), 2.45 (s, 1H), 2.41 (s, 2H), 1.99 (d, J = 13.0 Hz, 2H), 1.92 - 1.77 (m, 2H), 1.74 - 1.68 (m, 2H), 1.66 (s, 6H), 1.60 - 1.47 (m, 2H).

[0502] MS m / z (ESI): 404.3 [M+H] +

[0503] Example 11: 1-(2,2-difluoroethyl)-6-(6-((3-fluorobicyclo[1.1.1]pentan-1-yl)methoxy)-2- azaspiro[3.3]heptan-2-yl)-1H-pyrazolo[3,4-b]pyrazine

[0504] Reference Example 1 was synthesized according to the synthetic method of Reference Example 2, by replacing the compound 1-1 in Step 1 with (3-fluorobicyclo[1.1.1]pentane-1-carboxylic acid) was prepared according to the same method.

[0505] 1 H NMR (400 MHz, DMSO-d6) δ 8.13 (s, 1H), 7.88 (s, 1H), 6.43 (tt, J = 54.9, 3.8 Hz, 1H), 4.66 (td, J = 15.0, 3.8 Hz, 2H), 4.17 (s, 2H), 4.12 (s, 2H), 3.97 - 3.86 (m, 1H), 3.52 (s, 2H), 2.57 - 2.52 (m, 2H), 2.16 - 2.04 (m, 2H), 1.97 (d, J = 2.7 Hz, 6H).

[0506] MS m / z (ESI): 394.2 [M+H] +

[0507] Example 12: 1-(2,2-difluoroethyl)-6-(4-((3-fluorobicyclo[1.1.1]pentan-1-yl)methyl)piperazin-1- yl)-1H-pyrazolo[3,4-b]pyrazine

[0508] Reference Example 3 was synthesized according to the synthetic method of Reference Example 2, by replacing the compound 1-1 in Step 1 with (tert-butyl 2,7-diazaspiro[3.5]nonane-7-carboxylate) while replacing compound 3-1 with (3-fluorobicyclo[l. l. l ]pentane- l -carboxylic acid), compound 12 was prepared in the same manner.

[0509] 1 H NMR (400 MHz, DMSO-d6) δ 8.44 (s, 1H), 8.14 (s, 1H), 6.43 (tt, J = 54.8, 3.8 Hz, 1H), 4.69 (td, J = 15.0, 3.8 Hz, 2H), 3.73 (t, J = 5.0 Hz, 4H), 2.67 (s, 2H), 2.54 (t, J = 5.1 Hz, 4H), 2.02 (d, J = 2.7 Hz, 6H).

[0510] MS m / z (ESI): 367.2 [M+H] +

[0511] Example 13: l-(2,2-difluoroethyl)-6-(4-((l-methyl-2-oxabicyclo[2.1.1]hexan-4- yl)methyl)piperazin-l-yl)-lH-pyrazolo[3,4-b]pyrazine

[0512] Reference Example 3 was prepared in the same manner as described in Reference Example 1, replacing tert-butyl 2,7-diazaspiro[3.5]nonane-7-carboxylate in Step 1 with (tert-butyl 2,7-diazaspiro[3.5]nonane-7-carboxylate) while replacing compound 3-1 with (tert-butyl 2,7-diazaspiro[3.5]nonane-7-carboxylate) while replacing compound 3-1 with

[0513] 1 H NMR (400 MHz, DMSO-d6) δ 8.44 (s, 1H), 8.14 (s, 1H), 6.43 (tt, J = 54.8, 3.8 Hz, 1H), 4.69 (td, J = 15.0, 3.8 Hz, 2H), 3.73 (t, J = 5.0 Hz, 4H), 2.67 (s, 2H), 2.54 (t, J = 5.1 Hz, 4H), 2.02 (d, J = 2.7 Hz, 6H).

[0514] MS m / z (ESI): 367.2 [M+H] +

[0515] Example 14: l-(2,2-difluoroethyl)-6-(4-(l-methyl-2-oxabicyclo[2.1.1]hexan-4- yl)piperazin-l-yl)-lH-pyrazolo[3,4-b]pyrazine

[0516] Step 1: Synthesis of intermediate 14-2

[0517] Compound 14-1 (1.00 g) and 2,6-dimethylpyridine (1.05 g), sodium periodate (4.21 g) were dissolved in a mixed solution of ethyl acetate (10 mL) and water (10 mL), and potassium osmate (90.6 mg) was added at 0 °C. After stirring at room temperature for 3 hours, the reaction was completed. After separation, the ethyl acetate layer was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain crude intermediate 14-2 (1.16 g), which was directly used in the next reaction.

[0518] MS m / z (ESI): 236.1 [M+H] +

[0519] Step 2: Synthesis of 14-3

[0520] The intermediate 14-2 (1.16 g) and 1-methyl-2-oxabicyclo[2.1.1]hexan-4-amine hydrochloride (885 mg), triethylamine (2.49 g) in 10 mL of ethyl acetate were added, followed by sodium cyanoborohydride (1.55 g). After stirring at 20 °C for 16 hours, the reaction was complete. 20 mL of water was added, and the organic phase was extracted with ethyl acetate (20 mL*3), combined, and dried over anhydrous sodium sulfate. After filtration, the organic phase was concentrated under reduced pressure. The crude product was purified by flash silica gel column chromatography (40 g) (gradient 0-14% petroleum ether / tetrahydrofuran, flow rate 60 mL / min) to obtain intermediate 14-3 (500 mg). 40g flash silica gel column, gradient 0-14% petroleum ether / tetrahydrofuran, flow rate 60 mL / min).

[0521] MS m / z (ESI): 317.2 [M+H] +

[0522] Step 3: Synthesis of 14-4

[0523] Intermediate 14-3 (500 mg) was dissolved in 10 mL of tetrahydrofuran, and wet Pd / C (500 mg) was added. After stirring at 20 °C for 16 hours under a hydrogen atmosphere, the reaction was complete. After filtration, the reaction solution was concentrated under reduced pressure to obtain crude intermediate 14-4 (280 mg), which was directly used in the next reaction.

[0524] MS m / z (ESI): 183.2 [M+H] +

[0525] Step 4: Synthesis of compound 14

[0526] The intermediate 14-4 (180 mg) in the previous step and 6-chloro-l-(2,2- difluoroethyl)-lH-pyrazolo[3,4-b]pyrazine (215 mg) were dissolved in N,N- dimethylformamide (2 mL), sodium carbonate (314 mg) was added, and the reaction was completed after stirring at 100 °C for 16 hours. The reaction solution was concentrated under reduced pressure to obtain a crude product, which was purified by preparative high performance liquid chromatography (column: Welch Xtimate C18 150*25 mm*5 um; mobile phase: [acetonitrile-water (0.5‰ ammonia water + 0.01‰ ammonium bicarbonate)]; acetonitrile ratio: 13%-33%) to prepare compound 14 (25 mg).

[0527] MS m / z (ESI): 365.1 [M+H] + .

[0528] 1 H NMR (400 MHz, DMSO-d6) δ 8.47 (s, 1H), 8.15 (s, 1H), 6.45 (tt, J = 54.9, 3.8 Hz, 1H), 4.70 (td, J = 15.0, 3.8 Hz, 2H), 3.77 (t, J = 5.0 Hz, 4H), 3.59 (s, 2H), 2.66 (t, J = 5.1 Hz, 4H), 1.84 - 1.66 (m, 2H), 1.57 - 1.45 (m, 2H), 1.34 (s, 3H).

[0529] Example 15: 6-(4-(Bicyclo[l.l.l]pentan-l-yl)piperazin-l-yl)-l-(2,2- difluoroethyl)-lH-pyrazolo[3,4-b]pyrazine

[0530] Reference to the synthetic method of Example 14, 1-methyl-2-oxabicyclo[2.1.1]hexan-4-amine hydrochloride in Step 2 was replaced by Bicyclo[l.l.l]pentan-l-amine hydrochloride, and compound 15 was prepared by the same method.

[0531] 1 H NMR (400 MHz, DMSO-d6) δ 8.46 (s, 1H), 8.14 (s, 1H), 6.44 (tt, J = 54.8, 3.7 Hz, 1H), 4.69 (td, J = 15.0, 3.8 Hz, 2H), 3.75 (t, J = 5.1 Hz, 4H), 2.53 - 2.46 (m, 4H), 2.43 (s, 1H), 1.73 (s, 6H).

[0532] MS m / z (ESI): 335.2 [M+H] +

[0533] Example 16: l-(2,2-difluoroethyl)-6-(4-(3-fluorobicyclo[l. l. l]pentan-l- yl)piperazin-l-yl)-lH-pyrazolo[3,4-b]pyrazine

[0534] Reference to the synthesis method of Example 14, replace l-methyl-2-oxabicyclo[2.1.1]hexan-4-amine hydrochloride in Step 2 with (3-fluorobicyclo[l. l. l]pentan-l-amine hydrochloride), Compound 16 was obtained in the same manner.

[0535] 1 H NMR (400 MHz, DMSO-d6) δ 8.46 (s, 1H), 8.14 (s, 1H), 6.44 (tt, J = 54.8, 3.7 Hz, 1H), 4.69 (td, J = 15.0, 3.8 Hz, 2H), 3.80 - 3,60 (m, 4H), 2.53 - 2.46 (m, 4H), 2.02 (d, J = 2.7 Hz, 6H).

[0536] MS m / z (ESI): 353.2 [M+H] +

[0537] Example 17: (4-(l-(2,2-difluoroethyl)-lH-pyrazolo[3,4-b]pyrazin-6-yl)piperazin-l- yl)(3-fluorobicyclo[l. l. l]pentan-l-yl)methanone

[0538] Step 1: synthesis of intermediate 17-2

[0539] Compound 17-1 (1.00 g), piperazine-1-carboxylic acid tert-butyl ester (1.69 g) were dissolved in 20 mL of pyridine, l-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (3.42 g) was added at 0 °C, the reaction was completed after the reaction solution was stirred at 25 °C for 16 hours. The reaction solution was concentrated to obtain the crude product, which was purified by silica gel column chromatography (gradient 0-12% tetrahydrofuran / petroleum ether) to obtain intermediate 17-2 (1.80 g).

[0540] Step 2: synthesis of intermediate 17-3

[0541] Intermediate 17-2 (100 mg) was dissolved in 5 mL of dichloromethane, 1 mL of trifluoroacetic acid was slowly added dropwise at 0 °C, and the reaction was complete after stirring at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure to obtain the crude product of the title compound, intermediate 17-3, which was directly used in the next step.

[0542] MS m / z (ESI): 199.2 [M+H]+

[0543] Step 3: Synthesis of compound 17

[0544] The crude intermediate 17-3 (60 mg), 6-chloro-l-(2,2-difluoroethyl)-lH-pyrazolo[3,4- b]pyrazine (77.0 mg) were dissolved in N,N-dimethylformamide (2 mL), followed by the addition of sodium carbonate (102 mg). After stirring at 100 °C for 2 h, the reaction was completed. The reaction solution was cooled to room temperature, 30 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, and concentrated under reduced pressure. The crude product was separated and purified by high performance liquid chromatography (column: YMC TAR-C18, 30*150 mm, 5 um; mobile phase: [acetonitrile-water (0.5‰ ammonia water + 0.01‰ ammonium bicarbonate)]; acetonitrile ratio: 45-75%) to give compound 17 (108 mg).

[0545] 1 H NMR (400 MHz, Chloroform-d) d 8.23 (s, 1H), 8.08 (s, 1H), 6.21 (tt, J = 55.6, 4.5 Hz, 1H), 4.67 (td, J = 13.4, 4.4 Hz, 2H), 3.87-3.70 (m, 8H), 2.48 (d, J = 2.4 Hz, 6H).

[0546] MS m / z (ESI): 381.2 [M+H] +

[0547] Example 18: l-(2,2-difluoroethyl)-6-(2-(2-(trifluoromethyl)pyridin-4-yl)-2- azaspiro[3.5]nonan-7-yl)-l,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0548] 4-bromo-2-(trifluoromethyl)pyridine (56.8 mg), compound 10-5 (100 mg) and potassium carbonate (94.8 mg) were dissolved in N,N-dimethylformamide (2 mL). The reaction solution was stirred at 80 °C for 8 h, and then the reaction was completed. After the reaction solution was cooled to room temperature, 50 mL of water was added, and the mixture was extracted with ethyl acetate (10 mL*3). The combined organic phase was washed with 50 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by high performance liquid chromatography (column: YMC TAR-C18, 30*150 mm, 5 um; mobile phase: [acetonitrile-water (0.5‰ ammonia water + 0.01‰ ammonium bicarbonate)]; acetonitrile ratio: 60-90%) to give the title compound 18 (26.0 mg).

[0549] H NMR (400 MHz, Chloroform-d) δ 8.31 (d, J = 5.6 Hz, 1H), 7.99 (s, 1H), 7.89 (s, 1H), 6.62 (d, J = 2.3 Hz, 1H), 6.48 - 6.01 (m, 2H), 5.03 (td, J = 13.1, 4.4 Hz, 2H), 4.85 - 4.69 (m, 1H), 3.86 (s, 2H), 3.75 (s, 2H), 2.24 - 2.19 (m, 2H), 2.09 - 1.99 (m, 2H), 1.96 - 1.74 (m, 4H).

[0550] MS m / z (ESI): 469.2 [M+H] +

[0551] Example 19: l-(Bicyclo[l. l. l]pentan-l -yl)-4-(l -(2,2-difluoroethyl)-l H- pyrazolo[3,4-b]pyrazin-6-yl)piperazin-2-one

[0552] Reference Example 1 was followed, replacing intermediate 1-5 in Step 5 with ( l-(bicyclo[l. l. l]pentan-l -yl)piperazin-2-one) to give the title compound 19.

[0553] 1 H NMR (400 MHz, Chloroform-d) δ 8.15 (s, 1H), 8.09 (s, 1H), 6.21 (tt, J = 55.6, 4.5 Hz, 1H), 4.68 (td, J = 13.3, 4.5 Hz, 2H), 4.29 (s, 2H), 4.13 = 3.86 (m, 2H), 3.74 - 3.32 (m, 2H), 2.53 (s, 1H), 2.22 (s, 6H).

[0554] MS m / z (ESI): 349.1 [M+H] +

[0555] Example 20: 4-(l-(2,2-difluoroethyl)-lH-pyrazolo[3,4-b]pyrazin-6-yl)-l-(3- fluorobicyclo[l. l. l]pentan-l -yl)piperazin-2-one

[0556] Reference WO2016045587 for the synthesis of fragment BB-15, replacing BB-15-3 in Step 2 with (3-fluorobicyclo[l.l.l]pentan-l-amine hydrochloride) to give l-(3-fluorobicyclo[l.l.l]pentan-l-yl)piperazin-2-one (Compound 20-1).

[0557] Referring to the synthesis procedure of Example 1, intermediate 1-5 in Step 5 was replaced with Compound 20-1, and the title compound 20 was prepared in the same manner.

[0558] 1 H NMR (400 MHz, Chloroform-d) d 8.16 (s, 1H), 8.10 (s, 1H), 6.21 (tt, J = 55.6, 4.4 Hz, 1H), 4.68 (td, J = 13.4, 4.5 Hz, 2H), 4.33 (s, 2H), 4.18 - 3.96 (m, 2H), 3.69 - 3.32 (m, 2H), 2.54 (d, J = 2.1 Hz, 6H).

[0559] MS m / z (ESI): 367.1 [M+H] +

[0560] Example 21: l-(2,2-difluoroethyl)-6-(7-(2-(trifluoromethyl)pyridin-4-yl)-7- azaspiro[3.5]nonan-2-yl)-l,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0561] Referring to the synthesis procedure of Reference Example 10, tert-butyl 7-amino-2- azaspiro[3.5]nonane-2-carboxylate in Step 1 was replaced with (2-amino-7-azaspiro[3.5]nonane-7-carboxylic acid tert-butyl ester), and the title compound 21-5 was prepared in the same manner.

[0562] Referring to the synthesis procedure of Reference Example 18, Compound 10-5 was replaced with Compound 21-5, and the title compound 21 was prepared in the same manner.

[0563] 1H NMR (400 MHz, DMSO-d6) δ 8.34 (s, 1H), 8.24 (d, J = 5.9 Hz, 1H), 8.15 (s, 1H), 7.22 (d, J = 2.6 Hz, 1H), 7.12 - 7.03 (m, 1H), 6.43 (tt, J = 54.8, 3.8 Hz, 1H), 5.18 - 4.96 (m, 3H), 3.55 - 3.47 (m, 2H), 3.44 - 3.34 (m, 2H), 2.48 - 2.39 (m, 2H), 2.38 - 2.28 (m, 2H), 1.91 - 1.62 (m, 4H).

[0564] MS m / z (ESI): 469.1 [M+H] +

[0565] Example 22: 1-(2,2-difluoroethyl)-6-(2-(2-(trifluoromethyl)pyridin-4-yl)-2- azaspiro[3.3]heptan-6-yl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0566] Reference Example 10 was prepared according to the synthetic method of Reference Example 9, by replacing 7-amino-2-azaspiro[3.5]nonane-2-carboxylic acid tert-butyl ester in Step 1 with (6-amino-2-azaspiro[3.3]heptane-2-carboxylic acid tert-butyl ester) according to the same procedure to give compound 22-5.

[0567] Reference Example 18 was prepared according to the synthetic method of Reference Example 9, by replacing compound 10-5 with compound 22-5 to give the title compound 22.

[0568] 1 H NMR (400 MHz, DMSO-d6) δ 8.33 (s, 1H), 8.23 (d, J = 5.7 Hz, 1H), 8.15 (s, 1H), 6.74 (d, J = 2.2 Hz, 1H), 6.62 - 6.24 (m, 2H), 5.15 - 4.85 (m, 3H), 4.18 (s, 2H), 4.03 (s, 2H), 2.86 - 2.71 (m, 4H).

[0569] MS m / z (ESI): 441.1 [M+H] +

[0570] Example 23: 1-(2,2-difluoroethyl)-6-(2-(2-(trifluoromethyl)pyridin-4-yl)-2- azaspiro[4.5]dec-8-yl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0571] Reference to the synthesis method of Example 10, replace 7-amino-2-azaspiro[3.5]nonane-2- carboxylic acid tert-butyl ester in Step 1 with (8-amino-2-azaspiro[4.5]decane-2-carboxylic acid tert-butyl ester), the compound 23-5 was prepared in the same manner.

[0572] Reference to the synthesis method of Example 18, replace compound 10-5 with compound 23-5, the title compound 23 was prepared in the same manner.

[0573] 1 H NMR (400 MHz, DMSO-d6) δ 8.50 (s, 1H), 8.23 (d, J = 5.8 Hz, 1H), 8.16 (s, 1H), 6.90 (s, 1H), 6.71 (s, 1H), 6.63-6.18 (m, 1H), 5.16-4.95 (m, 2H), 4.79-4.73 (m, 1H), 3.48-3.36 (m, 4H), 2.11-1.70 (m, 8H), 1.69-1.51 (m, 2H).

[0574] MS m / z (ESI): 483.2 [M+H] +

[0575] Example 24: 1-(2,2-difluoroethyl)-6-(6-(2-(trifluoromethyl)pyridin-4-yl)-6-azaspiro[3.4]octan-2-yl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0576] Step 1: synthesis of intermediate 24-2

[0577] Compound 24-1 (250 mg) and methyl 1-(2,2-difluoroethyl)-4-nitro-1H-pyrazole-3-carboxylate (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 was complete after the reaction solution was stirred at 60 °C overnight. The reaction solution was cooled to room temperature, then 50 mL of water was added, followed by extraction with ethyl acetate (20 mL*3), the organic phase was combined and washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was separated and purified by column chromatography (petroleum ether / ethyl acetate: 2 / 1; Rf: 0.6) to obtain compound 24-2 (400 mg).

[0578] Step 2: synthesis of intermediate 24-3

[0579] Compound 24-2 (390 mg) was dissolved in a mixture solvent of ethanol / water (5 mL; 4:1), then iron powder (253 mg) and ammonium chloride (486 mg) were added. The reaction was completed after the reaction solution was stirred at 80°C for 2 hours. After the reaction solution was cooled to room temperature, it was filtered, the filtrate was added to 30 mL of water, 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 compound 24-3 (350 mg).

[0580] Step 3: Synthesis of intermediate 24-4

[0581] Compound 24-3 (350 mg) was dissolved in 4 mL of formamide, heated to 120°C and stirred for 48 hours. After the reaction was completed, the reaction solution was cooled to room temperature, added to 50 mL of water, extracted with ethyl acetate (20 mL*3), the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was separated and purified by column chromatography (petroleum ether / ethyl acetate: 1 / 1; Rf: 0.5) to obtain compound 24-4 (250 mg).

[0582] Step 4: Synthesis of intermediate 24-5

[0583] Compound 24-4 (250 mg) was dissolved in 10 mL of dichloromethane, 1.5 mL of trifluoroacetic acid was slowly added 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 compound 24-5 (250 mg).

[0584] Step 5: Synthesis of compound 24

[0585] 4-bromo-2-(trifluoromethyl)pyridine (146 mg), compound 24-5 (250 mg), and potassium carbonate (245 mg) were dissolved in N,N-dimethylformamide (3 mL), heated to 80°C and stirred for 6 hours. After the reaction was completed, the reaction solution was cooled to room temperature, added to 50 mL of water, extracted with ethyl acetate (20 mL*3), the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The concentrate was separated and purified by column chromatography (dichloromethane / methanol: 50 / 1; Rf: 0.6) and high performance liquid chromatography (column: YMC TAR-C18, 30*150 mm, 5 um; mobile phase: [water (0.5‰ ammonia water + 0.01‰ ammonium bicarbonate)-acetonitrile]; acetonitrile gradient: 40-70%) to obtain compound 24 (150 mg).

[0586] 1H NMR (400 MHz, DMSO-d6) δ 8.42 - 8.19 (m, 2H), 8.15 (s, 1H), 6.93 - 6.60 (m, 2H), 6.43 (tt, J=54.8, 3.7 Hz, 1H), 5.25 - 4.94 (m, 3H), 3.63 - 3.36 (m, 4H), 2.74 - 2.47 (m, 4H), 2.25 - 2.09 (m, 2H).

[0587] MS m / z (ESI): 455.2 [M+H] +

[0588] Example 25: 1-(2,2-difluoroethyl)-6-(2-(2-(trifluoromethyl)pyridin-4-yl)octahydrocyclopenta[c]pyrrol-5-yl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0589] Reference Example 24: 1-(2,2-difluoroethyl)-6-(2-(2-(trifluoromethyl)pyridin-4-yl)octahydrocyclopenta[c]pyrrol-5-yl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one was synthesized according to the procedure of Reference Example 23, substituting 24-1 in Step 1 with (5-aminooctahydro-2H-isoindole-2-carboxylic acid tert-butyl ester) according to the same procedure to give the title compound 25.

[0590] 1 H NMR (400 MHz, DMSO-d6) δ 8.42 - 8.13 (m, 3H), 6.96 - 6.67 (m, 2H), 6.66 - 6.26 (m, 1H), 5.32 - 4.96 (m, 3H), 3.75 - 3.24 (m, 4H), 3.25 - 2.80 (m, 2H), 2.39 - 2.26 (m, 2H), 2.17 - 1.90 (m, 2H).

[0591] MS m / z (ESI): 455.2 [M+H] +

[0592] Example 26: 1-(2,2-difluoroethyl)-6-(2-(2-(trifluoromethyl)pyridin-4-yl)octahydro-1H-isoindol-5-yl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0593] Reference Example 24: 1-(2,2-difluoroethyl)-6-(2-(2-(trifluoromethyl)pyridin-4-yl)octahydrocyclopenta[c]pyrrol-5-yl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one was synthesized according to the procedure of Reference Example 23, substituting 24-1 in Step 1 with (5-aminooctahydro-2H-isoindole-2-carboxylic acid tert-butyl ester) according to the same procedure to give the title compound 25.

[0594] 1H NMR (400 MHz, DMSO-d6) δ 8.40 (s, 1H), 8.21 (d, J = 5.8 Hz, 1H), 8.12 (s, 1H), 6.81 (s, 1H), 6.65 (s, 1H), 6.43 (tt, J = 54.9, 3.8 Hz, 1H), 5.04 (td, J = 14.6, 3.9 Hz, 2H), 4.75 - 4.71 (m, 1H), 3.70 - 3.36 (m, 4H), 2.59 - 2.51 (m, 2H), 2.21 - 1.58 (m, 6H).

[0595] MS m / z (ESI): 469.2 [M+H] +

[0596] Example 27: 1-(2,2-difluoroethyl)-6-(2-(2-(trifluoromethyl)pyridin-4-yl)-2- azaspiro[3.4]octan-6-yl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0597] Reference Example 24, Step 1 was replaced by (6-amino-2-azaspiro[3.4]octane-2-carboxylic acid tert-butyl ester) was prepared in the same manner as the title compound 27.

[0598] 1 H NMR (400 MHz, DMSO-d6) δ 8.29 (s, 1H), 8.23 (d, J = 5.7 Hz, 1H), 8.15 (s, 1H), 6.74 (d, J = 2.2 Hz, 1H), 6.65 - 6.25 (m, 2H), 5.16 - 4.96 (m, 3H), 4.11 - 3.98 (m, 2H), 3.98 - 3.88 (m, 2H), 2.45 - 1.95 (m, 6H).

[0599] MS m / z (ESI): 455.2 [M+H] +

[0600] Example 28: 1-(2,2-difluoroethyl)-6-(2-(2-(trifluoromethyl)pyridin-4-yl)-2- azaspiro[4.4]nonan-7-yl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0601] Reference Example 24, Step 1 was replaced by (7-amino-2-azaspiro[4.4]nonane-2-carboxylic acid tert-butyl ester) was prepared in the same manner as the title compound 28.

[0602] Example 29: 1-(2,2-difluoroethyl)-6-(6-(2-(trifluoromethyl)pyridin-4-yl)-6- azaspiro[3.5]nonan-2-yl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0603] Reference Example 24: Synthesis of 6-(6-(2-(trifluoromethyl)pyridin-4-yl)-6- azaspiro[3.5]nonan-2-yl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one (2-amino-6-azaspiro[3.5]nonane-6-carboxylic acid tert-butyl ester) was prepared in the same manner as described above.

[0604] 1 H NMR (400 MHz, CDC13) δ 8.39 - 8.29 (m, 1H), 8.01 - 7.88 (m, 2H), 7.13 - 7.01 (m, 1H), 6.88 - 6.65 (m, 1H), 6.42 - 6.01 (m, 1H), 5.27 - 4.85 (m, 3H), 3.51 - 3.43 (m, 2H), 3.41 - 3.30 (m, 2H), 2.65 - 2.17 (m, 4H), 1.96 - 1.70 (m, 4H).

[0605] MS m / z (ESI): 469.2 [M+H] +

[0606] Example 30: 1-(2,2-difluoroethyl)-6-(8-(2-(trifluoromethyl)pyridin-4-yl)-5-oxa-8- azaspiro[3.5]nonan-2-yl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0607] The synthetic route of compound 30 is as follows:

[0608] Example 31: 1-(2,2-difluoroethyl)-6-(7-oxo-6-(2-(trifluoromethyl)pyridin-4-yl)-6- azaspiro[3.4]octan-2-yl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0609] Step 1: Synthesis of intermediate 31-2

[0610] To a solution of compound 31-2 (240 mg) in 5 mL of dichloromethane was added trifluoroacetic acid (2.00 mL) dropwise at 0 °C. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure to give compound 31-3 (170 mg).

[0611] Step 2: Synthesis of compound 31-3

[0612] To a solution of compound 31-2 (240 mg) in 5 mL of dichloromethane was added trifluoroacetic acid (2.00 mL) dropwise at 0 °C. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure to give compound 31-3 (170 mg).

[0613] Step 3: Synthesis of intermediate 31-4

[0614] To a solution of compound 31-3 (160 mg) and 1-(2,2-difluoroethyl)-4-nitro-pyrazole-3- carboxylic acid methyl ester (198 mg) in 3 mL of tetrahydrofuran was added 1,8- diazabicyclo[5.4.0]undec-7-ene (426 mg) at 0 °C. The reaction mixture was stirred at 60 °C for 8 hours. The reaction mixture was cooled to room temperature and then added to 50 mL of water. 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, and the residue was purified by column chromatography (petroleum ether / ethyl acetate: 2 / 1; Rf: 0.6) to give compound 31-4 (70.0 mg).

[0615] Step 4: Synthesis of intermediate 31-5

[0616] Compound 31-4 (70.0 mg) was dissolved in a mixture solvent of ethanol / water (5 mL; 4:1), then iron powder (40.0 mg) and ammonium chloride (76.7 mg) were added. After stirring at 80 °C for 2 h, the reaction was completed. The reaction solution was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure, then 20 mL of water was added, extracted with ethyl acetate (50 mL*3), the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated to give compound 31-5 (60.0 mg).

[0617] Step 5: synthesis of compound 31

[0618] Compound 31-5 (50.0 mg) was dissolved in 1 mL of formamide. After stirring at 150 °C for 4 h, the reaction was completed. After the reaction solution was cooled to room temperature, 20 mL of water was added, then extracted with ethyl acetate (10 mL*3), the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was separated and purified by column chromatography-high performance liquid chromatography (column: YMC TAR-C18, 30*150 mm, 5um; mobile phase: [water-acetonitrile]; acetonitrile gradient: 25-45%) to give the title compound 31 (25.0 mg).

[0619] 1 H NMR (400 MHz, DMSO-d6) δ 8.70 (d, J = 5.6 Hz, 1H), 8.37 (d, J = 9.3 Hz, 1H), 8.29-8.17 (m, 1H), 8.16 (s, 1H), 7.99-7.77 (m, 1H), 6.71-6.21 (m, 1H), 5.32-4.86 (m, 3H), 4.12 (d, J = 55.6 Hz, 2H), 2.90 (d, J = 52.4 Hz, 2H), 2.80-2.57 (m, 4H).

[0620] MS m / z (ESI): 469.2 [M+H] +

[0621] Synthetic route of intermediate 31-1:

[0622] Example 32: 1-(2,2-difluoroethyl)-6-(2-((2-(trifluoromethyl)pyridin-4-yl)methyl)-2- azaspiro[3.5]nonan-7-yl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0623] To a solution of compound 10-5 (130 mg) in N,N-dimethylformamide (2.00 mL) was added 4-(bromomethyl)-2-(trifluoromethyl)pyridine (71.3 mg) and potassium carbonate (123 mg) at room temperature. The reaction was stirred at room temperature for 2 hours. The reaction was completed. The reaction mixture was added to 50 mL of water, and then 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. The residue was separated and purified by high performance liquid chromatography (column: YMC TAR-C18, 30*150 mm, 5um; mobile phase: [acetonitrile: water (0.5‰ ammonia water + 0.01‰ ammonium bicarbonate)]; acetonitrile gradient: 40-70%) to give the title compound 32 (40.0 mg).

[0624] 1 H NMR (400 MHz, DMSO-d6) δ 8.70 (d, J = 5.0 Hz, 1H), 8.30 (s, 1H), 8.12 (s, 1H), 7.77 (s, 1H), 7.63 (d, J = 5.0 Hz, 1H), 6.42 (tt, J = 54.8, 3.8 Hz, 1H), 5.03 (td, J = 14.6, 3.8 Hz, 2H), 4.70-4.53 (m, 1H), 3.77 (s, 2H), 3.10 (s, 2H), 3.00 (s, 2H), 2.05 (d, J = 13.0 Hz, 2H), 1.93-1.52 (m, 6H).

[0625] MS m / z (ESI): 483.1 [M+H] +

[0626] Example 33: 2-(1-(2,2-difluoroethyl)-1H-pyrazolo[3,4-b]pyrazin-6-yl)-7-((2- (trifluoromethyl)pyridin-4-yl)oxy)-5-oxa-2-azaspiro[3.4]octane

[0627] Step 1: Synthesis of intermediate 33-2

[0628] Compound 33-1 (0.90 g) was dissolved in dimethyl sulfoxide (20 mL) under nitrogen protection, and sodium hydride (314 mg) was added portionwise at 0 °C. The reaction was stirred at room temperature for 1 hour. Then 2-trifluoromethyl-4-bromopyridine (0.97 g) was added to the reaction solution, and the reaction was stirred for 2 hours. The reaction was completed. The reaction was quenched by adding saturated aqueous ammonium chloride solution, and then extracted with ethyl acetate (30 mL*3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was separated and purified by column chromatography (dichloromethane / methanol: 10 / 1; Rf: 0.6) to give compound 33-2 (1.10 g).

[0629] Step 2: Synthesis of intermediate 33-3

[0630] Compound 33-2 (0.20 g) was added to a solution of hydrogen chloride (10 mL, 4 M in ethyl acetate) under nitrogen protection. The reaction was complete after the reaction solution was stirred at room temperature for 2 hours under nitrogen protection. Compound 33-3 (0.15 g) was obtained after the reaction solution was concentrated under reduced pressure.

[0631] Step 3: Synthesis of compound 33

[0632] Compound 33-3 (100 mg), 6-chloro-l-(2,2-difluoroethyl)-lH-pyrazolo[3,4- b]pyrazine (97.7 mg) were dissolved in N,N-dimethylformamide (3.00 mL), followed by the addition of potassium carbonate (115 mg). The reaction was complete after the reaction solution was stirred at 90 °C for 2 hours. The reaction solution was cooled to room temperature, 10 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, and concentrated under reduced pressure. The residue was purified by column chromatography (petroleum ether / ethyl acetate: 20 / 1; Rf: 0.6) to give the title compound 33 (56.0 mg).

[0633] 1 H NMR (400 MHz, DMSO-d6) d 8.60 (d, J = 5.7 Hz, 1H), 8.15 (s, 1H), 7.95 (s, 1H), 7.46 (d, J = 2.4 Hz, 1H), 7.35 - 7.29 (m, 1H), 6.61 - 6.26 (m, 1H), 5.44 - 5.37 (m, 1H), 4.74 - 4.61 (m, 2H), 4.39 - 4.14 (m, 5H), 4.03 (d, J = 10.7 Hz, 1H), 2.61 - 2.52 (m, 2H).

[0634] MS m / z (ESI): 457.2 [M+H] +

[0635] Example 34: l-(2,2-difluoroethyl)-6-(5-(2-(trifluoromethyl)pyridin-4-yl)-4,5,6,7- tetrahydropyrazolo[l,5-a]pyrazin-2-yl)-l,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0636] Reference Example 24 was synthesized according to the procedure of Reference Example 23, replacing 24-1 in Step 1 with (2-amino-6,7-dihydropyrazolo[l,5-a]pyrazine-5(4H)-carboxylic acid tert-butyl ester) was prepared according to the same procedure.

[0637] Examples 35-43

[0638] The synthesis method of Reference Example 24 was referred to, except that 4-bromo-2- (trifluoromethyl)pyridine in step 5 was replaced by the starting material in the following table, and the following compounds 35-43 were synthesized in the same manner:

[0639] Example 44: 1-(2,2-difluoroethyl)-6-(6-(2-(trifluoromethyl)isonicotinoyl)-6- azaspiro[3.4]octan-2-yl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0640] Compound 24-5 (30.0 mg) and 2-(trifluoromethyl)isonicotinic acid (18.5 mg) were dissolved in N,N-dimethylformamide (2 mL), 2-(7-azabenzotriazol-1-yl)-N,N,N',N'- tetramethyluronium hexafluorophosphate (55.3 mg), N,N-diisopropylethylamine (37.6 mg) were added, and the reaction was completed after stirring at room temperature for 16 hours. 10 mL of water was added, and extraction was performed 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. The residue was separated and purified by preparative high-performance liquid chromatography (column: YMC TAR-C18, 30*150 mm, 5um; mobile phase: [water-acetonitrile]; acetonitrile gradient: 25-45%) to obtain the title compound 44 (11.7 mg).

[0641] 1 H NMR (400 MHz, DMSO-d6) δ 8.91-8.87 (m, 1H), 8.38-8.29 (m, 1H), 8.15-8.12 (m, 1H), 8.04-7.98 (m 1H), 7.91-7.71 (m, 1H), 6.61-6.19 (m, 1H), 5.22-4.91 (m, 3H), 3.60-3.35 (m, 4H), 2.67-2.59 (m, 2H), 2.47-2.30 (m, 2H), 2.15-1.94 (m, 2H).

[0642] MS m / z (ESI): 483.1 [M+H] +

[0643] Example 45: 2-(1-(2,2-difluoroethyl)-7-oxo-1,7-dihydro-6H-pyrazolo[4,3-d]pyrimidin-6-yl)- 7-(2-(trifluoromethyl)pyridin-4-yl)-5-oxa-7-azaspiro[3.4]octan-6-one

[0644] Step 1: Synthesis of intermediate 45-2

[0645] To a solution of 4-bromo-2-(trifluoromethyl)pyridine (280 mg), compound 45-1 (200 mg), tris(dibenzylideneacetone)dipalladium (75.6 mg), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (95.5 mg) and cesium carbonate (807 mg) in 1,4-dioxane (10 mL) was heated to 100 °C under nitrogen protection for 6 hours. The reaction was cooled to room temperature, 50 mL 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 and concentrated under reduced pressure. The residue was separated and purified by column chromatography (dichloromethane / methanol: 50 / 1; Rf: 0.5) to give compound 45-2 (240 mg). The title compound 45 was prepared by the same method as that of compound 31, steps 2-5.

[0646] 1 H NMR (400 MHz, DMSO-d6) δ 8.71 (t, J = 6.0 Hz, 1H), 8.31 (d, J = 2.7 Hz, 1H), 8.17 (s, 1H), 8.14-8.02 (m, 1H), 7.83-7.65 (m, 1H), 6.68-6.22 (m, 1H), 5.26-4.69 (m, 3H), 4.44 (s, 1H), 4.31 (s, 1H), 3.24-3.07 (m, 2H), 3.05-2.92 (m, 2H).

[0647] MS m / z (ESI): 471.0 [M+H] +

[0648] Synthetic route of compound 45-1:

[0649] Example 46: 1-(2,2-difluoroethyl)-6-(6-(2-(trifluoromethyl)thiazol-5-yl)-6- azaspiro[3.4]octan-2-yl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0650] Compound 24-5 (50.0 mg) and 2-(trifluoromethyl)-5-bromothiazole (30.8 mg) were dissolved in N,N-dimethylformamide (2 mL), and tris(dibenzylideneacetone)dipalladium (5.0 mg), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (10.0 mg) and cesium carbonate (60.0 mg) were added. The reaction was heated to 80 °C under nitrogen protection. After 5 hours, the reaction was completed. The reaction solution was filtered and concentrated, and the title compound 46 (4.00 mg) was obtained by preparative high performance liquid chromatography (column: YMC TAR-C18, 30*150 mm, 5 um; mobile phase: [water-acetonitrile]; acetonitrile gradient: 25-45%).

[0651] 1 H NMR (400 MHz, DMSO-d6) δ 8.37 (s, 1H), 8.15 (s, 1H), 6.89 (s, 1H), 6.57-6.29 (m, 1H), 5.22-4.96 (m, 3H), 3.60-3.30 (m, 4H), 2.81-2.60 (m, 2H), 2.37-1.86 (m, 4H).

[0652] MS m / z (ESI): 461.2 [M+H] + .

[0653] Example 47: 1-(2,2-difluoroethyl)-6-((3aR,5r,6aS)-2-(2-(trifluoromethyl)pyridin-4- yl)octahydrocyclopenta[c]pyrrol-5-yl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0654] Reference to the synthesis procedure of Example 24, the starting material compound 24-1 was replaced by compound 47-1, and compound 47 was prepared according to the above technical route.

[0655] 1 H NMR (400 MHz, DMSO-d6) δ 8.37 (s, 1H), 8.15 (s, 1H), 6.89 (s, 1H), 6.57-6.29 (m, 1H), 5.22-4.96 (m, 3H), 3.60-3.30 (m, 4H), 2.81-2.60 (m, 2H), 2.37-1.86 (m, 4H).

[0656] MS m / z (ESI): 461.2 [M+H] +

[0657] Example 48: 1-(2,2-difluoroethyl)-6-((3aR,5s,6aS)-2-(2-(trifluoromethyl)pyridin-4- yl)octahydrocyclopenta[c]pyrrol-5-yl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0658] Reference Example 24 was followed using compound 48-1 in place of compound 24-1 to give compound 48.

[0659] 1 H NMR (400 MHz, DMSO-d6) δ 8.32 (s, 1H), 8.25 (d, J = 5.8 Hz, 1H), 8.14 (s, 1H), 6.89 (d, J = 2.3 Hz, 1H), 6.82-6.70 (m, 1H), 6.60-6.18 (m, 1H), 5.31-5.20 (m, 1H), 5.11-4.97 (m, 2H), 3.70-3.53 (m, 2H), 3.31-3.07 (m, 4H), 2.41-2.27 (m, 2H), 2.22-1.98 (m, 2H).

[0660] MS m / z (ESI): 455.1 [M+H] +

[0661] Example 49: 1-(2,2-difluoroethyl)-5-methyl-6-(6-(2-(trifluoromethyl)pyridin-4-yl)-6- azaspiro[3.4]octan-2-yl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0662] Reference Example 24 was followed using compound 49-1 in place of compound 24-1 to give compound 49. The title compound 49 was obtained by the same method as described in Reference Example 24 using triethyl orthoformate

[0663] 1 H NMR (400 MHz, MeOD) δ 8.20 (d, J = 6.9 Hz, 1H), 7.85 (s, 1H), 7.18 (d, J = 2.4 Hz, 1H), 6.93 (d, J = 5.0 Hz, 1H), 6.28 (tt, J = 55.4, 4.2 Hz, 1H), 5.19-4.96 (m, 3H), 3.79 (s, 2H), 3.66 (t, J = 6.8 Hz, 2H), 3.46-3.32 (m, 2H), 2.64 (s, 3H), 2.56-2.44 (m, 2H), 2.31 (t, J = 6.8 Hz, 2H). ​

[0664] MS m / z (ESI): 469.3 [M+H] +

[0665] Example 50: 1-(Oxetan-3-yl)-6-(6-(2-(trifluoromethyl)pyridin-4-yl)-6- azaspiro[3.4]octan-2-yl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0666] Synthesis of Intermediate 50-4:

[0667] Step 1: Synthesis of Intermediate 50-2:

[0668] 4-nitro pyrazole (2.00 g) and cesium carbonate (11.5 g) were dissolved in N,N- dimethylformamide (15 mL), 3-iodooxetane (4.88 g) was added dropwise at 0 °C, the reaction was completed after stirring at room temperature for 2 hours. 100 mL of water was added, then extracted with ethyl acetate (100 mL*3), the organic phase was combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was separated and purified by column chromatography (petroleum ether / ethyl acetate: 10 / 1) to give the title compound 50-2 (2.50 g).

[0669] 1 H NMR (400 MHz, MeOD) δ 9.03 (d, J = 0.7 Hz, 1H), 8.41 (s, 1H), 5.65 (s, 1H), 5.00-4.83 (m, 4H).

[0670] MS m / z (ESI): 170.1 [M+H] +

[0671] Step 2: Synthesis of Intermediate 50-3:

[0672] Under nitrogen protection, intermediate 50-2 (2.30 g), n-pentyl chloroformate (3.07 g) were dissolved in methyl tert-butyl ether (30 mL), lithium bis(trimethylsilyl)amide in tetrahydrofuran (1 M, 16.2 mL) was added dropwise at -20 °C, the reaction was completed after stirring at -20 °C for 2 hours. The reaction solution was quenched with 30 mL of saturated ammonium chloride, then extracted with ethyl acetate (50 mL*3), the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was separated and purified by column chromatography (petroleum ether / ethyl acetate: 5 / 1) to give the title compound 50-3 (3.20 g).

[0673] 1H NMR (400 MHz, DMSO-d6) δ 8.54 (s, 1H), 5.84-5.76 (m, 1H), 4.98-4.89 (m, 4H), 4.37 (t, J = 6.6 Hz, 2H), 1.75-1.65 (m, 2H), 1.40-1.29 (m, 4H), 0.94-0.85 (m, 3H).

[0674] Step 3: Synthesis of intermediate 50-4:

[0675] Under nitrogen protection, intermediate 50-3 (3.00 g) was added to a mixed solution of tetrahydrofuran (20 mL) and water (10 mL), and lithium hydroxide (1.27 g) was added. After stirring at room temperature for 2 hours, the reaction was completed. The pH was adjusted to 6 with 2M hydrochloric acid solution, and the organic phase was extracted with ethyl acetate (100 mL*3), washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was swelled with ethyl acetate to obtain the target compound 50-4 (1.70 g).

[0676] 1 H NMR (400 MHz, DMSO-d6) δ 8.54 (s, 1H), 5.84-5.76 (m, 1H), 4.98-4.89 (m, 4H), 4.37 (t, J = 6.6 Hz, 2H), 1.75-1.65 (m, 2H), 1.40-1.29 (m, 4H), 0.94-0.85 (m, 3H).

[0677] Synthesis of compound 50:

[0678] Step 1: Synthesis of intermediate 50-6:

[0679] Compound 50-5 (600 mg), 4-bromo-2-(trifluoromethyl)pyridine (660 mg), and potassium carbonate (500 mg) were added to N,N-dimethylformamide (5 mL). Heating to 80°C, the reaction was completed after 2 hours. 50 mL of water was added to the reaction solution, and then extracted with ethyl acetate (20 mL*3), and 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 gradient = 10:1 ~ 5:1) to obtain the title compound 50-6 (600 mg).

[0680] Step 2: Synthesis of intermediate 50-7:

[0681] Compound 50-6 (300 mg) was dissolved in 5 mL of dichloromethane, followed by the addition of hydrogen chloride in 1,4-dioxane (4M, 3.00 mL). The reaction was completed after stirring at room temperature for 2 hours. The reaction was then concentrated under reduced pressure to obtain the title compound 50-7 (200 mg), which was directly used in the next step.

[0682] Step 3: Synthesis of intermediate 50-8:

[0683] Compound 50-4 (200 mg), compound 50-7 (188 mg) and N,N- diisopropylethylamine (285 mg) were added to N,N-dimethylformamide (5 mL) under nitrogen protection. 2-(7-Azabenzotriazol-l-yl)-N,N,N',N'- tetramethyluronium hexafluorophosphate (420 mg) was added at 0 °C. The reaction was completed after the reaction solution was stirred at room temperature for 2 hours. Then 20 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, and concentrated under reduced pressure. The residue was separated and purified by column chromatography (dichloromethane / methanol: 10 / 1) to obtain the title compound 50-8 (240 mg).

[0684] 1 H NMR (400 MHz, DMSO-d6) δ 9.46-9.34 (m, 1H), 8.49 (s, 1H), 8.28-8.16 (m, 1H), 6.88-6.74 (m, 1H), 6.73-6.60 (m, 1H), 5.62-5.44 (m, 1H), 4.91 (d, J = 6.7 Hz, 4H), 4.49-4.36 (m, 1H), 3.48 (s, 1H), 3.42-3.33 (m, 3H), 2.47-2.35 (m, 2H), 2.13-2.00 (m, 4H).

[0685] MS m / z (ESI): 466.9 [M+H] +

[0686] Step 4: synthesis of intermediate 50-9:

[0687] Compound 50-8 (200 mg) was added to a mixture of ethanol (8.0 mL) and water (2.0 mL) under nitrogen protection, and then iron powder (119 mg) and ammonium chloride (229 mg) were added. The reaction was completed after stirring at 70 °C for 2 hours. The reaction solution was cooled to room temperature, and the iron powder was removed by filtration. The filtrate was extracted with ethyl acetate (30 mL*3), and the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was separated and purified by column chromatography (dichloromethane / methanol: 10 / 1) to obtain the title compound 50-9 (130 mg).

[0688] 1H NMR (400 MHz, DMSO-d6) δ 8.24-8.19 (m, 1H), 8.19-8.10 (m, 1H), 7.25 (s, 1H), 6.88-6.75 (m, 1H), 6.72-6.58 (m, 1H), 5.89-5.80 (m, 1H), 4.88-4.77 (m, 4H), 4.53-4.47 (m, 2H), 4.47-4.38 (m, 1H), 3.50-3.44 (m, 1H), 3.44-3.34 (m, 2H), 2.39-2.29 (m, 2H), 2.16-2.00 (m, 4H).

[0689] MS m / z (ESI): 437.0 [M+H] +

[0690] Step 5: synthesis of compound 50:

[0691] Compound 50-9 (120 mg) was added to 10 mL formamide under nitrogen protection, the reaction was complete after the reaction solution was stirred at 150 °C for 2 hours. The reaction solution was cooled to room temperature, 10 mL was added, 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 separated and 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: 60%-90%) to obtain compound 50 (33.8 mg).

[0692] 1 H NMR (400 MHz, DMSO-d6) δ 8.35 (d, J = 5.8 Hz, 1H), 8.26-8.18 (m, 2H), 7.08-6.53 (m, 2H), 6.37-6.18 (m, 1H), 5.27-5.05 (m, 1H), 4.97 (d, J = 6.9 Hz, 4H), 3.65-3.35 (m, 6H), 2.71-2.55 (m, 2H), 2.31-2.08 (m, 2H).

[0693] MS m / z (ESI): 446.9 [M+H] +

[0694] Example 51: 1-(oxetan-3-yl)-6-((3aR,5r,6aS)-2-(2-(trifluoromethyl)pyridin-4-yl)octahydrocyclopenta[c]pyrrol-5-yl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0695] The synthesis method of Reference Example 50 was referred to, and the compound 50-5 in Step 1 was replaced with compound 51-1 The title compound 51 was prepared in the same manner.

[0696] 1 H NMR (400 MHz, DMSO-d6) δ 8.34 (s, 1H), 8.24 (d, J = 5.8 Hz, 1H), 8.19 (s, 1H), 6.88 (d, J = 2.4 Hz, 1H), 6.81 - 6.68 (m, 1H), 6.37 - 6.21 (m, 1H), 5.19 - 5.02 (m, 1H), 4.97 (d, J = 6.9 Hz, 4H), 3.69 - 3.54 (m, 2H), 3.54 - 3.44 (m, 2H), 3.03 - 2.83 (m, 2H), 2.44 - 2.22 (m, 2H), 2.08 - 1.93 (m, 2H).

[0697] MS m / z (ESI): 447.1 [M+H] +

[0698] Example 52: 1-(oxetan-3-yl)-6-(7-(2-(trifluoromethyl)pyridin-4-yl)-7-azaspiro[3.5]nonan-2-yl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0699] The synthesis method of Reference Example 50 was referred to, and the compound 50-5 in Step 1 was replaced with compound 52-1 The title compound 52 was prepared in the same manner.

[0700] 1 H NMR (400 MHz, DMSO-d6) δ 8.34 (s, 1H), 8.24 (d, J = 5.8 Hz, 1H), 8.19 (s, 1H), 6.88 (d, J = 2.4 Hz, 1H), 6.81 - 6.68 (m, 1H), 6.37 - 6.21 (m, 1H), 5.19 - 5.02 (m, 1H), 4.97 (d, J = 6.9 Hz, 4H), 3.69 - 3.54 (m, 2H), 3.54 - 3.44 (m, 2H), 3.03 - 2.83 (m, 2H), 2.44 - 2.22 (m, 2H), 2.08 - 1.93 (m, 2H).

[0701] MS m / z (ESI): 447.1 [M+H] +

[0702] Example 53: 1-(2,2-difluoroethyl)-6-(4-((6-(trifluoromethyl)pyridin-2-yl)methyl)piperazin-1-yl)-1H-pyrazolo[3,4-b]pyrazine

[0703] Step 1: Synthesis of intermediate 53-2

[0704] Piperazine-1-carboxylic acid tert-butyl ester (2.00 g) and 6-chloro-1-(2,2- difluoroethyl)pyrazolo[3,4-b]pyrazine (2.35 g) were dissolved in N,N-dimethylformamide (25 mL), sodium carbonate (3.41 g) was added, and the reaction was completed after stirring at 100 °C for 2 hours. The reaction was cooled to room temperature, 100 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, and concentrated under reduced pressure. The residue was separated and purified by column chromatography (petroleum ether / ethyl acetate: 10 / 1; Rf: 0.6) to give the title compound 53-2 (3.90 g).

[0705] Step 2: Synthesis of intermediate 53-3

[0706] Compound 53-2 (3.90 g) was dissolved in 20 mL of dichloromethane, followed by the addition of hydrogen chloride in 1,4-dioxane (4 M, 26.47 mL). The reaction was completed after stirring at room temperature for 2 hours. The reaction was concentrated under reduced pressure, slurried with ethyl acetate, filtered, and the filter cake was washed with ethyl acetate and then dried to give the title compound 53-3 (3.20 g).

[0707] Step 3: Synthesis of compound 53

[0708] Compound 53-3 (60.0 mg), 2-(bromomethyl)-6-(trifluoromethyl)pyridine (47.3 mg), and potassium carbonate (81.6 mg) were added to N,N-dimethylformamide (1.00 mL). The reaction was completed after stirring at room temperature for 2 hours. To the reaction was added 50 mL of water, and then 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. The residue was separated and purified by column chromatography (dichloromethane / methanol: 50 / 1; Rf: 0.6) and high performance liquid chromatography (column: YMC TAR-C18, 30*150 mm, 5 um; mobile phase: [water (containing 7 mmol / L ammonium bicarbonate)-acetonitrile]; acetonitrile gradient: 45-80%) to give compound 53 (38.0 mg).

[0709] 1H NMR (400 MHz, DMSO-d6) δ 8.46 (s, 1H), 8.27-8.01 (m, 2H), 7.83 (t, J = 8.0 Hz, 2H), 6.44 (tt, J = 54.9, 3.9 Hz, 1H), 4.70 (td, J = 15.0, 3.8 Hz, 2H), 3.85-3.75 (m, 6H), 2.75-2.56 (m, 4H).

[0710] MS m / z (ESI): 428.1 [M+H] +

[0711] Examples 54-62

[0712] Referring to the synthesis method of Example 53, the 2-(bromomethyl)-6- (trifluoromethyl)pyridine in Step 3 is replaced with the starting material in the following table, and the following compounds 54-62 are synthesized by the same method.

[0713] Example 63: 1-(2,2-difluoroethyl)-6-((3aR,5r,6aS)-2-(1-methyl-1H-pyrazol-4-yl) octahydrocyclopenta[c]pyrrol-5-yl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7- one

[0714] Compound 47-5 (100 mg), 4-bromo-1-methyl-1H-pyrazole (104 mg), Gphos G6 Pd (30.5 mg) and sodium trimethylsilanolate (95.4 mg) were dissolved in tetrahydrofuran (2 mL), the reaction solution was replaced with nitrogen three times, and after stirring at 90°C for 10 hours, the reaction was completed. 10 mL of water was added to the reaction solution, 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 separated and purified by column chromatography (dichloromethane / methanol: 10 / 1; Rf: 0.6) and high performance liquid chromatography (column: YMC TAR-C18, 30*150mm, 5um; mobile phase: [water (containing 7mmol / L ammonium bicarbonate)-acetonitrile]; acetonitrile gradient: 45-75%) to obtain compound 63 (25.0 mg).

[0715] 1H NMR (400 MHz, DMSO-d6) δ 8.28 (s, 1H), 8.13 (s, 1H), 7.17 (s, 1H), 7.05 (s, 1H), 6.44 (tt, J = 54.8, 3.8 Hz, 1H), 5.29 - 4.96 (m, 2H), 4.94 - 4.75 (m, 1H), 3.73 (s, 3H), 3.05 (d, J = 8.8 Hz, 2H), 2.89 - 2.68 (m, 4H), 2.42 - 2.22 (m, 2H), 2.10 - 1.78 (m, 2H).

[0716] MS m / z (ESI): 390.1 [M+H] +

[0717] Example 64: 1-(2,2-difluoroethyl)-6-((3aR,5r,6aS)-2-(1-methyl-2-oxo-1,2- dihydropyridin-4-yl)octahydrocyclopenta[c]pyrrol-5-yl)-1,6-dihydro-7H-pyrazolo[4,3- d]pyrimidin-7-one

[0718] The compound 64 was prepared according to the synthetic procedure of Reference Example 63, except that 4-bromo-1-methyl-1H-pyrazole in step was replaced by 4-bromo-1- methylpyridin-2(1H)-one, in the same manner.

[0719] 1 H NMR (400 MHz, DMSO-d6) δ 8.35 (s, 1H), 8.13 (s, 1H), 7.41 (d, J = 7.6 Hz, 1H), 6.70 - 6.23 (m, 1H), 5.84 (dd, J = 7.6, 2.4 Hz, 1H), 5.18 (d, J = 2.3 Hz, 1H), 5.11 - 4.93 (m, 3H), 3.61 - 3.43 (m, 2H), 3.39 - 3.29 (m, 2H), 3.27 (s, 3H), 2.90 - 2.84 (m, 2H), 2.40 - 2.23 (m, 2H), 2.03 - 1.93 (m, 2H).

[0720] Example 65: 1-(2,2-difluoroethyl)-6-((3aR,5r,6aS)-2-(1-(2,2,2-trifluoroethyl)-1H- pyrazol-4-yl)octahydrocyclopenta[c]pyrrol-5-yl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin- 7-one

[0721] Compound 65 was prepared according to the synthetic procedure of Reference Example 63, replacing 4-bromo-l-methyl-lH-pyrazole in Step with 4-bromo-l-(2,2,2-trifluoroethyl)-lH-pyrazole.

[0722] 1 H NMR (400 MHz, DMSO-d6) δ 8.30 (s, 1H), 8.13 (s, 1H), 7.39 - 7.17 (m, 2H), 6.44 (tt, J = 54.8, 3.8 Hz, 1H), 5.15 - 4.75 (m, 5H), 3.19 - 3.05 (m, 2H), 2.97 - 2.67 (m, 4H), 2.43 - 2.19 (m, 2H), 2.01 - 1.80 (m, 2H).

[0723] MS m / z (ESI): 458.1 [M+H] +

[0724] Example 66: l-(2,2-difluoroethyl)-6-((3aR,5r,6aS)-2-(l-methyl-3-(trifluoromethyl)-lH- pyrazol-4-yl)octahydrocyclopenta[c]pyrrol-5-yl)-l,6-dihydro-7H-pyrazolo[4,3- d]pyrimidin-7-one

[0725] Compound 66 was prepared according to the synthetic procedure of Reference Example 63, replacing 4-bromo-l-methyl-lH-pyrazole in Step with 4-bromo-l-methyl-3- (trifluoromethyl)-lH-pyrazole.

[0726] 1 H NMR (400 MHz, DMSO-d6) δ 8.23 (s, 1H), 8.14 (s, 1H), 7.55 (d, J = 1.3 Hz, 1H), 6.44 (tt, J = 54.8, 3.8 Hz, 1H), 5.14 - 4.98 (m, 2H), 4.96 - 4.79 (m, 1H), 3.83 (s, 3H), 3.08 - 2.98 (m, 2H), 2.98 - 2.86 (m, 2H), 2.85 - 2.65 (m, 2H), 2.43 - 2.27 (m, 2H), 1.98 - 1.75 (m, 2H).

[0727] MS m / z (ESI): 458.1 [M+H] +

[0728] Example 67: 1-(2,2-difluoroethyl)-6-((3aR,5r,6aS)-2-(1-methyl-5-(trifluoromethyl)-1H- pyrazol-4-yl)octahydrocyclopenta[c]pyrrol-5-yl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7- one

[0729] Compound 67 was prepared according to the procedure of Reference Example 63, replacing 4-bromo-1-methyl-1H-pyrazole in Step 1 with 4-bromo-1-methyl-5-(trifluoromethyl)-1H- pyrazole.

[0730] 1 H NMR (400 MHz, DMSO-d6) d 8.17 (s, 1H), 8.09 (s, 1H), 7.36 (s, 1H), 6.38 (tt, J = 54.8, 3.8 Hz, 1H), 5.05 - 4.92 (m, 2H), 4.92 - 4.77 (m, 1H), 3.85 (d, J = 1.3 Hz, 3H), 3.07 - 2.90 (m, 4H), 2.68 - 2.65 (m, 2H), 2.37 - 2.18 (m, 2H), 1.92 - 1.69 (m, 2H).

[0731] MS m / z (ESI): 458.0 [M+H] +

[0732] Example 68: 1-(2,2-difluoroethyl)-6-(7-((6-(trifluoromethyl)pyridin-2-yl)methyl)-7- azaspiro[3.5]nonan-2-yl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0733] Compound 21-5 (100 mg), 2-(bromomethyl)-6-(trifluoromethyl)pyridine (60.0 mg) and potassium carbonate (200 mg) were added into N,N-dimethylformamide (4.00 mL). The reaction was completed after the reaction solution was stirred at room temperature for 2 hours. 50 mL of water was added to the reaction solution, which was then 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. Compound 68 (38.0 mg) was obtained by separation and purification of the residue 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: 50-80%).

[0734] 1H NMR (400 MHz, DMSO-d6) δ 8.31 (s, 1H), 8.14 (s, 1H), 8.07 (t, J = 7.8 Hz, 1H), 7.86-7.72 (m, 2H), 6.42 (tt, J = 54.8, 3.8 Hz, 1H), 5.15-4.95 (m, 3H), 3.65 (s, 2H), 2.46-2.18 (m, 8H), 1.81-1.54 (m, 4H).

[0735] MS m / z (ESI): 483.1 [M+H] +

[0736] Examples 69-77

[0737] Referring to the synthesis method of Example 68, the 2-(bromomethyl)-6- (trifluoromethyl)pyridine in the reaction was replaced with the starting material in the following table, and the following compounds 69-77 were synthesized by the same method.

[0738] Example 78: 1-(2,2-difluoroethyl)-6-((3aR,5r,6aS)-2-((6-(trifluoromethyl)pyridin-2- yl)methyl)octahydrocyclopenta[c]pyrrol-5-yl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin- 7-one

[0739] Compound 47-5 (70.5 mg), 2-(bromomethyl)-6-(trifluoromethyl)pyridine (40.0 mg) and potassium carbonate (150 mg) were added to N,N-dimethylformamide (2.00 mL). After the reaction was completed by stirring the reaction liquid at room temperature for 2 hours, 50 mL of water was added to the reaction liquid, which was then 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. Compound 78 (20.0 mg) was obtained by separating and purifying the residue by high performance liquid chromatography (column: YMC TAR-C18, 30*150 mm, 5um; mobile phase: [water (containing 7 mmol / L ammonium bicarbonate)-acetonitrile]; acetonitrile gradient: 50-80%).

[0740] 1H NMR (400 MHz, DMSO-d6) δ 8.33 (s, 1H), 8.15 (s, 1H), 8.09 (t, J = 7.8 Hz, 1H), 7.96 (d, J = 7.9 Hz, 1H), 7.78 (d, J = 7.6 Hz, 1H), 6.44 (tt, J = 54.8, 3.8 Hz, 1H), 5.05 (td, J = 14.6, 3.8 Hz, 2H), 4.96 - 4.73 (m, 1H), 3.83 (s, 2H), 2.77 - 2.56 (m, 4H), 2.41 - 2.19 (m, 4H), 1.96 - 1.78 (m, 2H).

[0741] MS m / z (ESI): 469.0 [M+H] +

[0742] Examples 79-89

[0743] Referring to the synthetic method of Example 78, the following compounds 79-89 were synthesized by replacing 2-(bromomethyl)-6-(trifluoromethyl)pyridine with the starting materials in the table below.

[0744] Example 90: 4-(2-(1-(2,2-difluoroethyl)-7-oxo-1,7-dihydro-6H-pyrazolo[4,3- d]pyrimidin-6-yl)-6-azaspiro[3.4]octan-6-yl)-2-(trifluoromethyl)benzonitrile:

[0745] Compound 24-5 (100 mg), 4-bromo-2-(trifluoromethyl)benzonitrile (100 mg), Xphos Pd G4 (20.0 mg) and cesium carbonate (200 mg) were dissolved in 1,4-dioxane (2 mL), the reaction solution was replaced with nitrogen three times, and after stirring at 80 °C for 16 hours, the reaction was completed. 10 mL of water was added to the reaction solution, 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 separated and purified by column chromatography (dichloromethane / methanol: 10 / 1; Rf: 0.6) and high performance liquid chromatography (column: YMC TAR-C18, 30*150mm, 5um; mobile phase: [water (containing 7mmol / L ammonium bicarbonate)-acetonitrile]; acetonitrile gradient: 45-75%) to obtain compound 90 (20.6 mg).

[0746] 1H NMR (400 MHz, DMSO-d6) δ 8.38 (d, J = 6.7 Hz, 1H), 8.15 (s, 1H), 7.80 (d, J = 8.3 Hz, 1H), 7.04 - 6.72 (m, 2H), 6.57 - 6.29 (m, 1H), 5.23 - 4.95 (m, 3H), 3.73 - 3.36 (m, 4H), 2.74 - 2.57 (m, 4H), 2.26 - 2.07 (m, 2H).

[0747] MS m / z (ESI): 479.3 [M+H] +

[0748] Example 91: 1-(2,2-difluoroethyl)-6-((3aR,5r,6aS)-2-(2-(2-hydroxypropan-2- yl)pyridin-4-yl)octahydrocyclopenta[c]pyrrol-5-yl)-1,6-dihydro-7H-pyrazolo[4,3- d]pyrimidin-7-one

[0749] Compound 91 was prepared according to the synthetic procedure of Reference Example 63, replacing 4-bromo-1-methyl-1H-pyrazole in Step with 2-(4-bromopyridin-2- yl)propan-2-ol.

[0750] 1H NMR (400 MHz, MeOD-d4) δ 8.56 (br s, 1H), 8.21 (s, 1H), 8.02 (d, J = 7.0 Hz, 1H), 7.98 (s, 1H), 6.90 - 6.71 (m, 2H), 6.27 (tt, J = 55.4, 4.1 Hz, 1H), 5.27 - 4.96 (m, 3H), 4.09 - 3.62 (m, 4H), 3.18 - 2.95 (m, 2H), 2.59 - 2.37 (m, 2H), 2.34 - 2.17 (m, 2H), 1.62 (s, 6H).

[0751] MS m / z (ESI): 445.2 [M+H] +

[0752] Example 92: 6-((3aR,5r,6aS)-2-(2-cyclopropylpyridin-4-yl)octahydrocyclopenta[c]pyrrol-5- yl)-1-(2,2-difluoroethyl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0753] Compound 92 was prepared according to the synthetic procedure of Reference Example 63, replacing 4-bromo-1-methyl-1H-pyrazole in Step with 4-bromo-2-cyclopropylpyridine.

[0754] 1 H NMR (400 MHz, DMSO) δ 8.35 (s, 1H), 8.13 (s, 1H), 7.96 (d, J = 5.9 Hz, 1H), 6.71 - 6.20 (m, 3H), 5.24 - 4.73 (m, 3H), 3.58 - 3.26 (m, 4H), 2.97 - 2.73 (m, 2H), 2.42 - 2.21 (m, 2H), 2.20 - 1.84 (m, 3H), 1.04 - 0.76 (m, 4H).

[0755] MS m / z (ESI): 427.2 [M+H] +

[0756] Example 93: 1-(2,2-difluoroethyl)-6-((3aR,5r,6aS)-2-(2-morpholino pyridin-4-yl) octahydrocyclopenta[c]pyrrol-5-yl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0757] Reference Example 63 was synthesized according to the procedure of Reference Example 62, substituting 4-(4-bromopyridin-2-yl)morpholine for 4-bromo-1-methyl-1H-pyrazole in Step 1. Compound 93 was prepared in the same manner.

[0758] 1 H NMR (400 MHz, DMSO-d6) δ 8.37 (s, 1H), 8.14 (s, 1H), 7.75 (d, J = 7.3 Hz, 1H), 6.64 - 6.22 (m, 2H), 5.92 (d, J = 1.8 Hz, 1H), 5.21 - 4.89 (m, 3H), 3.86 - 3.56 (m, 8H), 3.53 - 3.44 (m, 4H), 2.93 (br s, 2H), 2.39 - 2.22 (m, 2H), 2.12 - 1.94 (m, 2H).

[0759] MS m / z (ESI): 472.1 [M+H] +

[0760] Example 94: 1-(2,2-difluoroethyl)-6-((3aR,5s,6aS)-2-((1-methyl-3-(trifluoromethyl)-1H-pyrazol-4-yl)methyl)octahydrocyclopenta[c]pyrrol-5-yl)-1,6-dihydro-7H-pyrazolo[4,3-d]pyrimidin-7-one

[0761] Compound 48-5 (80.0 mg), 4-(bromomethyl)-1-methyl-3-(trifluoromethyl)-1H-pyrazole (40.0 mg) and potassium carbonate (150 mg) were added into N,N-dimethylformamide (2.00 mL). The reaction was completed after the reaction solution was stirred at room temperature for 2 hours. 50 mL of water was added to the reaction solution, which was 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 separated and 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: 50-80%) to obtain compound 94 (23.0 mg).

[0762] 1 H NMR (400 MHz, DMSO-d6) δ 8.19 (s, 1H), 8.15 (s, 1H), 7.91 (s, 1H), 6.57-6.30 (m, 1H), 5.16-4.97 (m, 2H), 4.90-4.75 (m, 1H), 3.89 (s, 3H), 3.53 (s, 2H), 2.74-2.55 (m, 4H), 2.32-2.24 (m, 4H), 1.67-1.66 (m, 2H).

[0763] MS m / z (ESI): 472.1 [M+H] +

[0764] Biological test method

[0765] Experimental Example 1: Assay of the GCase enzyme activity of the compound of the present disclosure on living lysosomes of HEK-293T cell line

[0766] 1) Material reagent:

[0767] 2) Instrument:

[0768] 3) Experimental method:

[0769] Day 1: Plating

[0770] Inoculate 2500 HEK-293T cells per well in a 96-well plate, 100 μL / well DMEM medium (containing 10% FBS), and incubate at 37°C in a 5% carbon dioxide environment for 48 hours.

[0771] Day 3: Drug addition

[0772] Prepare different concentrations of the compound of the present disclosure (compound stock solution of 10 mM prepared with DMSO, diluted with DMEM medium solution to 1 μM, 5 μM, 10 μM, respectively). After discarding the old culture medium of the 96-well plate, add the medium mixed with 1 μM, 5 μM, 10 μM of the compound to the 96-well plate at a volume of 200 μL / well, and place in a 37°C, 5% CO2 incubator for 72 hours;

[0773] Day 6: enzyme activity detection

[0774] a) Preparation of mixed medium of PFB-FDGlu: Take 22 mL of phenol red-free DMEM medium, add 22 μL of PFB-FDGlu (50 μg / mL) and mix well;

[0775] b) Preparation of mixed medium of PFB-FDGlu and Bafilomycin A1: Take 11 mL of the prepared mixed medium of PFB-FDGlu above into a new 15 mL tube, add 2.2 μL of Bafilomycin A1 and mix well, ready for use;

[0776] c) Discard the culture medium, wash the cells once with 100 μL / well of phenol red-free DMEM medium, and add the prepared mixed medium containing only PFB-FDGlu and the mixed medium containing PFB-FDGlu and Bafilomycin A1 (100 μL / well), respectively;

[0777] d) Place in a 37°C incubator for 1 hour of reaction incubation;

[0778] e) After incubation, perform kinetic curve reading with a microplate reader at 15 min / time for 2 hours (excitation wavelength 485 nm / emission wavelength 525 nm). Calculate AC 50 (refer to the drug concentration when the GCase enzyme activation activity is 150%).

[0779] The test results of the compound of the present disclosure on the activation level of GCase enzyme in HEK-293T cells are shown in Table 1.

[0780] Table 1 Test results of the compound of the present disclosure on GCase enzyme activity in HEK293T cells

[0781] Experimental Example 2: Inhibition of CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP3A4 enzyme activity by the compound of the present disclosure

[0782] The reagents and consumables not specifically mentioned in this test are commercially available.

[0783] The inhibition of CYP1A2, CYP2C9, CYP2C19, CYP2D6, CYP3A4 enzyme activity of the compounds of the present disclosure was determined using the following test method.

[0784] The test procedure is as follows:

[0785] 1. Preparation of 100 mM phosphate buffer (PBS): Weigh 7.098 g of Na2HPO4, add 500 mL of pure water and ultrasonically dissolve as solution A. Weigh 3.400 g of KH2PO4, add 250 mL of pure water and ultrasonically dissolve as solution B. Place the A solution on a stirrer and slowly add the B solution until the pH value reaches 7.4 to prepare a 100 mM PBS buffer.

[0786] 2. Preparation of NADPH solution: Prepare a 10 mM NADPH (reduced coenzyme II) solution with 100 mM PBS buffer.

[0787] 3. Preparation of test compound working solution: Dilute the 10 mM DMSO stock solution of the compounds of the present disclosure with DMSO to obtain the compound working solution (10000 μM, 3333.3 μM, 1111.1 μM, 370.37 μM, 123.46 μM, 41.15 μM, 13.7 μM, 0 μM).

[0788] 4. Preparation of positive inhibitor working solution: Dilute the positive inhibitor DMSO stock solution (CYP 2C9: Sulfaphenazole 1000 μM, CYP 2D6: Quinidine / CYP 3A4: Ketoconazole, 100 μM; CYP 1A2: α-naphthoflavone, 50 μM; CYP 2C19: Norketone, 6000 μM) with DMSO to obtain the positive inhibitor working solution (Sulfaphenazole, 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-naphthoflavone, 50 μM, 15 μM, 5 μM, 1.5 μM, 0.5 μM, 0.15 μM, 0 μM; Norketone, 6000 μM, 1800 μM, 600 μM, 180 μM, 60 μM, 18 μM, 0 μM).

[0789] 5. Preparation of substrate working solution: Prepare the substrate working solution (8000 μM Phenacetin, 1000 μM Diclofenac, 8000 μM S-mephenytoin, 1000 μM dextromethorphan and 400 μM midazolam) with water, acetonitrile or acetonitrile / methanol.

[0790] 6. Take 1 μL of 20 mg / mL liver microsomes solution (source: gentest), 1 μL of substrate working solution and 177 μL of PBS buffer, add 1 μL of compound working solution (1 μL of positive inhibitor working solution is added instead of compound working solution for positive control group), mix well, and pre-incubate in a 37°C water bath for 10 minutes. Pre-incubate 10 mM NADPH solution in a 37°C water bath for 10 minutes. After 10 minutes, add 20 μL of NADPH to each well to start the reaction, and incubate at 37°C for 15 minutes (CYP1A2), 15 minutes (CYP2C9), 60 minutes (CYP2C19), 15 minutes (CYP2D6), or 15 minutes (CYP3A4). All incubation samples are set in duplicate. After the corresponding incubation time, add 400 uL of ice methanol containing internal standard to all samples to terminate the reaction. Mix well, and centrifuge at 4000 rpm at 4°C for 30 minutes. After centrifugation, transfer 100 μL of supernatant to a sample injection plate, add 200 μL of ultrapure water, mix well, and use for LC-MS / MS analysis.

[0791] The values of IC50 of the compounds of the present disclosure for CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4 were calculated by Excel XLfit 5.3.1.3. 50 The values are shown in Table 2.

[0792] Table 2. IC50 values of the compounds of the present disclosure for CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4 50 The values are shown in Table 2. “-” indicates not detected

[0793] Experimental Example 3: High-content cell imaging assay for measuring the activity of lysosomes GCase enzyme in SH-SY5Y cell line

[0794] 1) Material reagents:

[0795] 2) Instruments:

[0796] 3) Experimental method:

[0797] Cell complete culture medium: DMEM / F-12, HEPES, containing 10% FBS, containing 1% penicillin-streptomycin;

[0798] High-content imaging detection culture medium: DMEM / F-12, HEPES, without phenol red, without FBS;

[0799] Inoculate 20,000 SH-SY5Y cells per well in 384-well plates with 40 μL / well of cell complete medium, and incubate at 37°C in a 5% CO2 environment overnight.

[0800] Dissolve the compounds of the present disclosure in DMSO to a concentration of 20 mM, and dilute to 10 mM with DMSO at a ratio of 1:1. Then dilute the 20 mM and 10 mM compound solutions with cell complete medium at a ratio of 1:400 to a concentration of 50 μM and 25 μM, respectively. Add 10 μL of medium containing the compounds of the present disclosure at a concentration of 50 μM or 25 μM to the cell wells of the 384-well plate, i.e., the final concentration of the test compound is 10 μM or 5 μM, and the final concentration of DMSO is 0.05%. Centrifuge at 500 rpm for 30 seconds, and incubate at 37°C in a 5% CO2 environment for 72 hours.

[0801] Prepare the high-content imaging detection medium (DMEM / F-12, HEPES, without phenol red, without FBS), and add Lysotracker at a final concentration of 75 nM and PFB-FDGlu at a final concentration of 58 μM; remove the original medium supernatant in the cell plate, and add the medium containing 40 μL of Lysotracker and PFB-FDGlu, centrifuge at 500 rpm for 30 seconds, and incubate at 37°C in a 5% CO2 environment for 45 minutes; remove the cell plate, and perform continuous high-content imaging at an interval of 30 minutes for a total time of 3 hours, collect images in the bright field, Alexa 488, and Alexa 647 channels, and take 10 fields of view for each well, and calculate the average fluorescence intensity of the cells, the average fluorescence intensity of the lysosomes, and the average fluorescence intensity of the cell-lysosome co-localization. Take the DMSO well without the drug as the background, calculate the fluorescence intensity at the end point of each well, and take the drug concentration when the fluorescence intensity of the test compound is twice the background fluorescence intensity as the AC 100 value (AC 100 , which refers to the drug concentration when the GCase enzyme activation activity is 200%).

[0802] The compounds of the present disclosure show high GCase activation levels in SH-SY5Y cells, and the test results are shown in Table 3. The test compounds have AC 100 values in the range of: A: AC 100 ≤ 5 μM; B: 5 μM < AC 100 ≤ 10 μM; C: > 10 μM;

[0803] Table 3 Test results of the compounds of the present disclosure on GCase enzyme activity in SH-SY5Y cells

[0804] Experimental Example 4: Determination of the potential inhibitory effect of the compounds of the present disclosure on the voltage-gated potassium ion channel hERG

[0805] The experiment selected a Chinese hamster ovary (CHO) cell (B'SYS GmbH) line stably expressing hERG potassium ion channel. The hERG-CHO cells were clamped by SyncroPatch 384i / 384 (Nanion) automatic patch clamp system to form a whole cell voltage clamp mode, and the hERG current was induced by the corresponding voltage. The test compound was tested: first, the cells were washed with extracellular fluid containing 0.1% DMSO for 6 times, and the stable hERG current was measured as the detection baseline, and the baseline current value was the average of 5 stable sampling points (tail current size 空白 ). After the hERG current remained stable, 5 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, and 10 minutes were waited for the compound to fully act on the cells and record the hERG current synchronously. After the current tends to be stable, 5 stable hERG current values are read, and their average is taken as the final current value at a specific concentration (tail current size 化合物 ). Cisapride (Sigma (C4740)) was used as a positive drug for a synchronous determination of 6 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 test results. After testing the compound, 450 nM dofetilide (Beijing Ipsen Technology Development Co., Ltd., D525700) was added to all test cells to completely inhibit their current, as a complete positive control of the cells (tail current size 阳性对照 ). Finally, the inhibition rate of the tail current was calculated by the following formula {tail current inhibition rate (%) = [1- (tail current size 化合物 -tail current size 阳性对照 ) / (tail current size 空白 -tail current size 阳性对照 )] x 100%}, the dose-effect curve fitting was performed by Graphpad Prism 8.0 software, and the IC 50 value was calculated.

[0806] The inhibition of the compounds of the present disclosure on hERG is shown in Table 4 below, and the test data shows that the hERG inhibition activity of the compounds of the present disclosure is poor, and the potential safety risk in the future is small.

[0807] Table 4 Inhibition activity of voltage-gated potassium ion channel hERG of the compounds of the present disclosure

Claims

A compound represented by Formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein is selected from a single bond or a double bond; Q is selected from N or C; G is selected from N, CH or C(O); Ring B is selected from the following groups: 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; or, b) W is selected from the group consisting of a bond, -CR a R b -, -CR a R b O-, C(O), O, or S; R a , R b is independently selected from H, halogen, Ci-C6alkyl, Ci-C6haloalkyl or Ci-C4alkoxy; Ring A is selected from C3-C 12 cycloalkyl, 4-12 membered heterocyclyl, 5-10 membered heteroaryl, or phenyl; Each R 1 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 the C1-C6 alkyl, C1-C6 alkoxy, C3-C6 cycloalkyl or 4-10 heterocyclic group is optionally substituted with NH2, OH, halogen or C1-C6 alkyl; R 2 is selected from H, C1-C6alkyl, C3-C6cycloalkyl, or 4-10 membered heterocyclyl, said C1-C6alkyl, C3-C6cycloalkyl, or 4-10 membered heterocyclyl optionally substituted with NH2, OH, halogen, C1-C6alkyl, C3-C6cycloalkyl, or 4-6 membered heterocyclyl; R 3 is selected from H, C1-C6alkyl, C1-C6alkoxy, C3-C6cycloalkyl, or 4-6 membered heterocyclyl, said C1-C6alkyl, C3-C6cycloalkyl, or 4-6 membered heterocyclyl optionally substituted with NH2, OH, halogen, or C1-C6alkyl; 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; R 5 selected from H, deuterium, NH2, CN, OH, halogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, C3-C6cycloalkyl, or 4-7 membered heterocyclyl; p, q are independently selected from 0, 1 or 2. The compound of Formula (I) according to claim 1, or a stereoisomer or a pharmaceutically acceptable salt thereof, wherein, wherein, is a single bond, Q is N and G is C(O); or is a double bond, Q is C and G is N. The compound of Formula (I) according to claim 1 or 2, or a stereoisomer or a pharmaceutically acceptable salt thereof, wherein, The compound represented by formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof is selected from the compound represented by formula (I-2) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof: ring A, ring B, R 1 , R 2 , R 3 , R 4 , R 5 , p, q, W are as defined in claim 1; or wherein: Ring B is selected from C6-C 14 Spirocycloalkylene, 6-14 membered spiroheterocyclic alkylene, or 6-14 membered fused heterocyclic alkylene; W is selected from the group consisting of a bond, -CR a R b -, -CR a R b O-, C(O) or O; R a , R b is independently selected from H or C1-C6 alkyl; Ring A is selected from C3-C 12 Cycloalkyl, 4-12 membered heterocyclic, 5-10 membered heteroaryl or phenyl; Each R 1 Independently selected from CN, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C3-C6 cycloalkyl or 4-10 membered heterocyclic group, wherein the C1-C6 alkyl and C1-C6 alkoxy are optionally substituted with OH or halogen; R 2 selected from C1-C6alkyl or 4-10 membered heterocyclyl, said C1-C6alkyl optionally substituted with halogen; R 3 selected from H, C1-C6 alkyl or C1-C6 alkoxy; R 4 selected from =0 or C1-C6alkyl; R 5 is selected from H or C1-C6 alkyl. The compound of Formula (I) or a stereoisomer or a pharmaceutically acceptable salt thereof according to any one of claims 1-3, wherein, Ring B is selected from the following groups: a) 6-14 membered spiro- heterocyclylenyl, 6-14 membered fused- heterocyclylenyl or 5-12 membered bridged- heterocyclylenyl; or, b) or Ring B is selected from the following groups: a) 6-14 membered spiro- heterocyclylenyl or 6-14 membered fused- heterocyclylenyl; or b) or Ring B is selected from 6-14 membered spiro- heterocyclylenyl or 6-14 membered fused- heterocyclylenyl; or Ring B is selected from 6-14 membered spiro- heterocyclylenyl, 6-14 membered fused- heterocyclylenyl or 5-12 membered bridged- heterocyclylenyl; or Ring B is selected from 3 / 6 bicyclic spiro- heterocyclylenyl, 4 / 4 bicyclic spiro- heterocyclylenyl, 4 / 5 bicyclic spiro- heterocyclylenyl, 4 / 6 bicyclic spiro- heterocyclylenyl, 5 / 5 bicyclic spiro- heterocyclylenyl or 5 / 6 bicyclic spiro- heterocyclylenyl; or Ring B is selected from 4 / 4 bicyclic spiro- heterocyclylenyl, 4 / 5 bicyclic spiro- heterocyclylenyl, 5 / 6 bicyclic spiro- heterocyclylenyl, 4 / 6 bicyclic spiro- heterocyclylenyl, 5 / 6 bicyclic fused- heterocyclylenyl or 5 / 5 bicyclic fused- heterocyclylenyl; or Ring B is selected from 4 / 4 bicyclic spiro- heterocyclylenyl, 4 / 5 bicyclic spiro- heterocyclylenyl, 5 / 6 bicyclic spiro- heterocyclylenyl or 4 / 6 bicyclic spiro- heterocyclylenyl; Alternatively, ring B is selected from Alternatively, ring B is selected from: Alternatively, ring B is selected from: The compound of Formula (I) or a stereoisomer or a pharmaceutically acceptable salt thereof according to any one of claims 1-4, wherein, W is selected from the group consisting of a bond, -CR a R b -, -CR a R b O- or C(O); or W is selected from a bond, -CR a R b -, -CR a R b O-, C(O) or O; or W is selected from a bond, -CR a R b - or -CR a R b O-; or W is selected from a bond or -CR a R b O-; or W is selected from -CR a R b - or -CR a R b O-; or W is selected from a bond, -CH2-, -CH2O- or C(O); or W is selected from a bond, -CH2- or -CH2O-; or W is selected from a bond or -CH2O-. The compound of Formula (I) or a stereoisomer or a pharmaceutically acceptable salt thereof according to any one of claims 1-5, wherein, Ring A is selected from C3-C 12 cycloalkyl, 4-12 membered heterocyclyl, or 5-10 membered heteroaryl; or, ring A is selected from C3-C 12 cycloalkyl, 5-10 membered heteroaryl, or phenyl; or, ring A is selected from C3-C 12 cycloalkyl or 5-10 membered heteroaryl; Alternatively, ring A is selected from C3-C6 cycloalkyl, 4-9 membered heterocyclic, 5-6 membered heteroaryl, or phenyl; or ring A is selected from C3-C6 cycloalkyl, 4-9 membered heterocyclic, 5-6 membered heteroaryl, or phenyl. 12 Cycloalkyl or 4-12 membered heterocyclic groups; or, ring A is selected from C5-C 10 bridged cycloalkyl, C5-C 10 spirocycloalkyl or 4-9 membered heterocyclyl; or, ring A is selected from C5-C 10 bridged cycloalkyl; or Ring A is selected from 5-7 membered heterocyclyl; or Ring A is 5-6 membered heteroaryl; Alternatively, ring A is selected from Alternatively, ring A is selected from Alternatively, ring A is selected from The compound of Formula (I) or a stereoisomer or a pharmaceutically acceptable salt thereof according to any one of claims 1-6, wherein, R 1 independently selected from CN, NH2, OH, halogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, C3-C6cycloalkyl, or 4-10 membered heterocyclyl, said C3-C6cycloalkyl or 4-10 membered heterocyclyl optionally substituted with NH2, OH, halogen, C1-C6alkyl; or R 1 independently selected from CN, halogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, C3-C6cycloalkyl, or 4-10 membered heterocyclyl, said C1-C6alkyl or C1-C6alkoxy optionally substituted with OH or halogen; or R is H, or R 1 independently selected from halogen, C1-C6alkyl, C1-C6haloalkyl, or C1-C6alkoxy optionally substituted with halogen; Or, R 1 Selected from C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 alkoxy, wherein the C1-C6 alkoxy group is optionally substituted with a halogen; or R 1 is selected from halogen, Ci-C6-alkyl or Ci-C6-haloalkyl. The compound of Formula (I) or a stereoisomer or a pharmaceutically acceptable salt thereof according to any one of claims 1-7, wherein, R 2 H, C1-C6 alkyl, C3-C6 cycloalkyl, or 4-10 membered heterocyclyl, said C1-C6 alkyl, C3-C6 cycloalkyl, or 4-10 membered heterocyclyl optionally substituted with NH2, OH, halogen, or C1-C6 alkyl; or R is selected from C1-C6alkyl, C3-C6cycloalkyl, or 4-10 membered heterocyclyl, said C1-C6alkyl, C3-C6cycloalkyl, or 4-10 membered heterocyclyl optionally substituted with NH2, OH, halo, or C1-C6alkyl; 2 or R is selected from C1-C6alkyl, C3-C6cycloalkyl, or 4-10 membered heterocyclyl, said C1-C6alkyl, C3-C6cycloalkyl, or 4-10 membered heterocyclyl optionally substituted with NH2, OH, halo, or C1-C6alkyl; or R 2 is selected from C1-C6alkyl, C3-C6cycloalkyl, or 4-6 membered heterocyclyl, said C1-C6alkyl, C3-C6cycloalkyl, or 4-6 membered heterocyclyl optionally substituted with NH2, OH, halogen, or C1-C6alkyl; or R 2 is selected from C1-C6alkyl or 4-10 membered heterocyclyl, said C1-C6alkyl being optionally substituted with halogen; or R 2 Ci-C6alkyl substituted by halogen; Or, R 2 Selected from CH2CHF2 or oxocyclic butyl; or R 2 is CH2CHF2. The compound of Formula (I) according to any one of claims 1-8, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein R 3 selected from H, C1-C6 alkyl or C1-C6 alkoxy; Or, R 3 It's H. The compound of Formula (I) according to any one of claims 1-9, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein, R 4 selected from deuterium, OH, halogen, C1-C6alkyl, C1-C6haloalkyl, C1-C6alkoxy, or C3-C6cycloalkyl; Or, R 4 Selected from deuterium, OH, halogen, =O, C1-C6 alkyl or C1-C6 haloalkyl; or R 4 is selected from =0 or C1-C6alkyl; or R 4 is =0. The compound of Formula (I) according to any one of claims 1-10, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein R 5 selected from H, deuterium, halogen or C3-C6cycloalkyl; or R 5 is selected from H, halogen, Ci-C6alkyl or C3-C6cycloalkyl; or R 5 is selected from H or Ci-C6alkyl; or R 5 is selected from H or CH3. The compound of Formula (I) according to any one of claims 1-11, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein, p is selected from 0 or 1; and / or q is selected from 0 or 1. The compound of Formula (I) or a stereoisomer or a pharmaceutically acceptable salt thereof according to any one of claims 1-12, wherein, The compound represented by formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof is selected from the compound represented by formula (II-2) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof: X1is selected from O, S or CR 6 R 7 ; X2is selected from CR 8 or N; X3is selected from C, 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, n are independently selected from 0, 1 or 2; q' is selected from 0 or 1; q" is selected from 0 or 1; Ring A, R 1 , R 2 , R 3 , R 4 , R 5 , p, W are defined as in any one of claims 1-12. The compound of Formula (I) according to any one of claims 1-12, or a stereoisomer or a pharmaceutically acceptable salt thereof, is selected from the group consisting of a compound of Formula (V-2), or a stereoisomer or a pharmaceutically acceptable salt thereof: Ring A, R 1 , R 2 , R 3 , R 4 , R 5 , p, q, W are defined as in any one of claims 1-12. The compound of Formula (I) according to any one of claims 1-12, or a stereoisomer or a pharmaceutically acceptable salt thereof, is selected from the group consisting of a compound of Formula (VI-2), or a stereoisomer or a pharmaceutically acceptable salt thereof: 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 are independently selected from 0, 1 or 2; q' is selected from 0 or 1; q" is selected from 0 or 1; Rings A, R 1 R 2 R 3 R 4 R 5 p, W are as defined in any one of claims 1 to 12. The compound of Formula (I) according to any one of claims 1-2, or a stereoisomer or a pharmaceutically acceptable salt thereof, is selected from the group consisting of a compound of Formula (I-1), or a stereoisomer or a pharmaceutically acceptable salt thereof: Ring A is selected from C3-C 12 cycloalkyl or 4-12 membered heterocyclyl; when W is selected from a bond or -CR a R b - when W is selected from a bond or -CR Ring B, R 1 , R 2 , R 3 , R 4 , R 5 , p, q, W are defined as in any of claims 1-2. The compound of Formula (I) according to any one of claims 1-2, or a stereoisomer or a pharmaceutically acceptable salt thereof, is selected from the group consisting of a compound of Formula (I-1), or a stereoisomer or a pharmaceutically acceptable salt thereof: Ring A is selected from C3-C6 cycloalkyl, 4-9 membered heterocyclyl or 5-6 membered heteroaryl; R 1 , R 2 , R 3 , R 4 , R 5 , p, q, W are defined as in any one of claims 1-2. The compound of Formula (I) according to any one of claims 1-3, or a stereoisomer or pharmaceutically acceptable salt thereof, wherein, The compound represented by formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof is selected from the following compounds or a stereoisomer thereof or a pharmaceutically acceptable salt thereof: The compound of Formula (I) according to claim 1 or 2, 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 following compounds or a stereoisomer thereof or a pharmaceutically acceptable salt thereof: A pharmaceutical composition comprising a compound of Formula (I) according to any one of claims 1-19, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. Use of a compound of Formula (I) according to any one of claims 1-19, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 20, for the preparation 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 subject, comprising administering to a subject, preferably a mammal, more 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 to 19, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described in claim 20. The use as described in claim 21 or the method as described in claim 22, wherein: the GCase-mediated disease is a disease that benefits from allosteric modulation of GCase; or the GCase-mediated disease is a disease that benefits from activation of GCase; or the GCase-mediated disease is selected from neurodegenerative diseases; preferably, the neurodegenerative disease is Alzheimer’s disease or Parkinson’s disease.

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