USP30 inhibitor compound, pharmaceutical composition, preparation method therefor, and use thereof

Abstract

The present invention provides a compound having USP30 inhibitor activity, a use thereof, a preparation method therefor, and a composition containing the inhibitor. These inhibitors are useful in a variety of therapeutic fields, including diseases or conditions involving mitochondrial dysfunction, such as PD, cancer, and fibrosis.

Description

USP30 inhibitor compounds, pharmaceutical compositions, their preparation methods and applications

[0001] This invention claims priority to two earlier applications: Patent Application No. 202411604161.1, filed with the China National Intellectual Property Administration on November 11, 2024, entitled "USP30 Inhibitor Compound, Pharmaceutical Composition and Preparation Method Thereof and Application Thereof"; and Patent Application No. 202510011964.4, filed with the China National Intellectual Property Administration on January 3, 2025, entitled "USP30 Inhibitor Compound, Pharmaceutical Composition and Preparation Method Thereof and Application Thereof". The entire contents of the aforementioned earlier applications are incorporated herein by reference. Technical Field

[0002] This invention belongs to the field of pharmaceutical compounds, specifically relating to a class of nitrile compounds with USP30 inhibitory activity, their uses, preparation methods, and compositions containing the inhibitor. Background Technology

[0003] Ubiquitination plays an important role in maintaining intracellular protein homeostasis. Damaged or misfolded proteins are recognized and ubiquitinated by specific E3 ubiquitin ligases. Ubiquitinated proteins then participate in different physiological processes within the cell, the most important of which is entering the proteasome for degradation and clearance. In addition to the ubiquitin-proteasome pathway, some ubiquitin-labeled proteins are also cleared through the autophagy-lysosome system. Damaged mitochondria are cleared and renewed through this pathway, a process known as mitophagy.

[0004] Mitophagy (mitochondrial autophagy) is an important mitochondrial regulatory mechanism. Under stress such as reactive oxygen species (ROS) exposure, mitochondrial DNA (mtDNA) mutations gradually accumulate, leading to decreased mitochondrial membrane potential and depolarization damage, ultimately resulting in cell death. To maintain mitochondrial and cellular homeostasis and prevent damaged mitochondria from harming cells, cells selectively encapsulate and degrade damaged or dysfunctional mitochondria through mitophagy. The main pathway is PINK & Parkin-mediated mitophagy. When mitochondria are damaged, PINK1 accumulates on the surface of the outer mitochondrial membrane (OMM) and is autophosphorylated and activated there. Activated PINK1 phosphorylates ubiquitin, thereby recruiting Parkin from the cytoplasm to the mitochondria. Then, PINK1 phosphorylates and activates phosphorylated ubiquitin-bound Parkin, enabling Parkin to ubiquitinate proteins on the outer mitochondrial membrane. Finally, damaged mitochondria are recognized, encapsulated, and transported to lysosomes for degradation by autophagosomes through the large amount of ubiquitinated proteins on their surface. USP30 is a deubiquitinizing enzyme and the only USP located on the mitochondrial membrane. Therefore, it is also the most important protein that antagonizes Parkin ubiquitination. By deubiquitinizing mitochondrial outer membrane proteins, it restricts mitophagy. Damaged mitochondria accumulate in the cell because they cannot be cleared in time, which leads to the corresponding damage to cell and tissue functions.

[0005] Mitochondrial diseases (MDs) are structural or functional disorders of mitochondria caused by mutations in mitochondrial DNA (mtDNA) and / or nuclear DNA (nDNA). They are common inherited metabolic disorders in childhood, with most having poor prognoses and being among the leading causes of disability and death in childhood. Currently, there are no effective treatments, thus necessitating the development of new drugs. Numerous preclinical and ex vivo studies based on patient cells have demonstrated that reduced mitophagy levels are a key characteristic of MDs, and enhancing mitophagy can improve disease symptoms. Inhibiting USP30 activation of the Parkin / PINK pathway to enhance autophagy may offer a new approach to the treatment of MDs. Furthermore, mitochondrial accumulation is also associated with neurodegenerative diseases such as Parkinson's disease (PD). Therefore, this invention aims to develop a highly efficient small molecule USP30 for the treatment of MDs and PD. Summary of the Invention

[0006] To address the aforementioned technical problems, the present invention provides a compound of formula (I), its racemic mixture, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt, or prodrug compound:

[0007] in,

[0008] X1, X2, X3, X4, and X5 may be the same or different, and are independently selected from CR1 or N;

[0009] Each R1 may be the same or different, and is independently selected from H, CN, halogen, unsubstituted, or optionally by one, two, or more Rs. c The following groups are substituted: OH, C 1-12 Alkyl, C 2-12 alkenyl, C 2-12 alkynyl, halogenated C 1-12 Alkyl, C 1-12 Alkoxy, C 3-12 Cycloalkyl, 3-14 membered heterocyclic groups, C 6-14 Aryl, 5-14 heteroaryl, -N(R) 11 (R) 12 -COR 13 , or -S(O)2R 14 ; Each R c They are either the same or different, and are independently selected from OH, -N(R) 15 (R) 16 CN, halogens, oxometalates (=O), C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 cycloalkyl or 3-6 membered heterocyclic groups; R 11 R 12 R 14 R 15 R 16 Whether the two are the same or different, they are selected independently from H and C. 1-6 Alkyl, Halogenated C 1-6 Alkyl or C 3-6 cycloalkyl; R 13 Selected from H, OH, -NH2, C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 cycloalkyl, 3-6 membered heterocyclic, C 6-10 Aryl or 5-10 heteroaryl groups;

[0010] Ring A is a 5-10 member heteroaryl group;

[0011] Each R a They may be identical or different, and are independently selected from CN, halogen, unsubstituted, or optionally composed of one, two, or more R. a1 The following groups are substituted: OH, C 1-12Alkyl, Halogenated C 1-12 Alkyl, C 1-12 Alkoxy, C 1-12 Alkylthio, C 3-12 Cycloalkyl, 3-14 membered heterocyclic groups or -NH2; each R a1 They are selected independently of OH, -NH2, CN, halogens, oxo (=O), and C, whether they are the same or different. 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl or 3-6 membered heterocyclic groups;

[0012] m is selected from 0, 1, 2, 3 or 4;

[0013] Y is absent or selected from CO, CONR2, or unsubstituted or optionally substituted by one, two or more R3 groups, including 5-14 member heteroaryl groups or CONR2-C. 1-12 Alkylene; R2 is selected from H, C 1-6 Alkyl or C 3-6 Cycloalkyl; each R3 may be the same or different, and is independently selected from CN, halogen, OH, NH2, C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1- 6-alkoxy, C 3-6 Cycloalkyl or 3-6 membered heterocyclic groups;

[0014] Ring B is a 3-14 member N-containing heterocyclic ring;

[0015] Each R b They may be identical or different, and are independently selected from CN, halogen, oxo (=O), unsubstituted, or optionally substituted by one, two, or more R. b1 The following groups are substituted: OH, C 1-12 Alkyl, C 2-12 alkenyl, C 2-12 alkynyl, halogenated C 1-12 Alkyl, C 1- 12 Alkoxy, C 1-12 Alkylthio, C 3-12 Cycloalkyl, 3-14 membered heterocyclic groups, C 6-14 Aryl, 5-14 heteroaryl, -N(R) 41 (R) 42 ) or -S(O)2R 43 Or, two R atoms attached to the same carbon atom b Together with the carbon atom it is attached to, it forms an unsubstituted or optionally substituted form with one, two or more R atoms. b1 The following ring systems are replaced: C3-14 A carbon ring or a 3-14 membered heterocycle; or, two R atoms attached to adjacent carbon atoms. b Together with the carbon atoms respectively attached thereto, they form unsubstituted or optionally substituted with one, two or more R atoms. b1 Substitution of the following groups: olefinic bond (C=C), C 3-14 Carbon rings, 3-14 membered heterocycles, C 6-14 Aromatic rings or 5-14 heterocyclic aromatic rings; R 41 R 42 R 43 Whether the two are the same or different, they are selected independently from H and C. 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 3-6 cycloalkyl; each R b1 They are selected independently of OH, -NH2, CN, halogens, oxo (=O), and C, whether they are the same or different. 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl or 3-6 membered heterocyclic groups;

[0016] n is selected from 0, 1, 2, 3, 4, or 5; and

[0017] At least one of rings A and B is a bicyclic (such as a fused ring, a bridged ring, or a spiral ring) system.

[0018] According to some implementation schemes, X1 is selected from CR1 or N; X2, X3, X4, and X5 are CR1.

[0019] According to some implementation schemes, each R1 may be the same or different, and is independently selected from H, CN, halogens (e.g., F, Cl, Br), OH, C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy (e.g., OCF3) or C 3-6 Cycloalkyl.

[0020] According to some implementation schemes, X1 is selected from CH, CF or N; X2 is selected from CH, COCF3 or CCN; X3, X4 and X5 are CH.

[0021] According to some implementation plans Selected from

[0022] According to some implementation schemes, ring A is selected from...

[0023] According to some implementation schemes, each R aWhether the same or different, they are independently selected from CN, halogen, and C. 1-6 Alkyl (e.g., methyl), halogenated C 1-6 Alkyl (e.g., CHF2), C 1-6 Alkoxy, halogenated C 1-6 Alkoxy or C 3-6 Cycloalkyl.

[0024] According to some implementation schemes, m is selected from 0 or 1.

[0025] According to some implementation plans Selected from The "*" side is connected to Y, and the "#" side is connected to Y.

[0026] According to some implementation schemes, Y is absent or selected from CO, CONR2, CONR2-C(R3)2- or a 5-membered heteroaryl group (e.g., ...). ).

[0027] According to some implementation schemes, R2 is selected from H or C. 1-3 alkyl.

[0028] According to some implementation schemes, each R3 may be the same or different, and is independently selected from H or C. 1-3 alkyl.

[0029] According to some implementation plans, ring B is selected from...

[0030] According to some implementation schemes, each R b They may be the same or different, and are independently selected from CN, halogen, oxo (=O), OH, -NH2, -S(O)2C 1-3 Alkyl, C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, C 3-6 Cycloalkyl, 3-6 membered heterocyclic, phenyl or 5-6 membered heteroaryl.

[0031] According to some implementation schemes, two R atoms attached to the same carbon atom b Together with the carbon atom it is attached to, it forms the following group: C 3-6 Carbon rings or 3-6 membered heterocycles; or, two R atoms attached to adjacent carbon atoms. b Together with the carbon atoms they are attached to, they form the following groups: olefinic bond (C=C), C 3-6 Carbon rings, 3-6 membered heterocyclic rings, benzene rings, or 5-6 membered heteroaromatic rings.

[0032] According to some implementation schemes, n is 0, 1, or 2.

[0033] According to some implementation plans Selected from

[0034] According to some embodiments, the compound represented by formula (I) has the following structure:

[0035] in,

[0036] X1 is selected from CR 1a Or N; R 1a Selected from H, halogens (e.g., F, Cl or Br), OCF3 or CN;

[0037] X2 is CR 1b Or N; R 1b Selected from H, halogens (e.g., F, Cl or Br), OCF3 or CN;

[0038] X6 is selected from CH or N;

[0039] X7 is selected from O, S, or NH;

[0040] R2 is selected from H or C. 1-3 alkyl;

[0041] n is 0 or 2;

[0042] When n is 2, two R atoms attached to the same carbon atom b Together with the carbon atom it is attached to, it forms the following group: C 3-6 Carbon rings or 3-6 membered heterocycles; or, two R atoms attached to adjacent carbon atoms. b Together with the carbon atoms they are attached to, they form the following groups: olefinic bond (C=C), C 3-6 Carbon rings, 3-6 membered heterocycles, benzene rings, or 5-6 membered heteroaromatic rings;

[0043] q1 is selected from 1 or 2;

[0044] q2 is selected from 0, 1 or 2.

[0045] According to some embodiments, the compound represented by formula (I) has the following structure:

[0046] in,

[0047] X1 is selected from CR 1a Or N; R 1a Selected from H, F, Cl, or Br;

[0048] X2 is CR 1b ;R 1b Selected from H, OCF3, or CN;

[0049] X7 is selected from O, S, or NH;

[0050] R2 is selected from H or C. 1-3 alkyl;

[0051] n is 2; and the two R atoms connected to the same carbon atom b Together with the carbon atom it is attached to, it forms the following group: C 3-6 Carbon rings or 3-6 membered heterocycles; or, two R atoms attached to adjacent carbon atoms. b Together with the carbon atoms they are attached to, they form the following groups: olefinic bond (C=C), C 3-6 Carbon rings, 3-6 membered heterocycles, benzene rings, or 5-6 membered heteroaromatic rings;

[0052] q1 is selected from 1 or 2;

[0053] q2 is selected from 0, 1 or 2.

[0054] According to some embodiments, the compound represented by formula (I) has the following structure:

[0055] in,

[0056] X1 is selected from CR 1a Or N; R 1a Selected from H, F, Cl, or Br;

[0057] X2 is CR 1b ;R 1b Selected from H, OCF3, or CN;

[0058] X7 is selected from O, S, or NH;

[0059] R2 is selected from H or C. 1-3 alkyl;

[0060] p is selected from 0 or 1;

[0061] Each R3 may be the same or different, and is independently selected from H or C. 1-3 alkyl;

[0062] n is selected from 0, 1, 2, or 3;

[0063] Each R b They are the same or different, and are independently selected from halogens and C. 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkyl, Halogenated C 1-3alkoxy group; or, two R groups attached to the same carbon atom. b Together with the carbon atom it is attached to, it forms the following group: C 3-6 Carbon rings or 3-6 membered heterocycles; or, two R atoms attached to adjacent carbon atoms. b Together with the carbon atoms they are attached to, they form the following groups: olefinic bond (C=C), C 3-6 Carbocyclic rings, 3-6 membered heterocyclic rings, benzene rings, or 5-6 membered heteroaromatic rings; or, R b R2, together with the atoms it is attached to, forms a 3-6 membered heterocycle;

[0064] q1 is selected from 1 or 2;

[0065] q2 is selected from 0, 1 or 2.

[0066] According to some embodiments, the compound shown in formula (I) is selected from the following structures:

[0067] The present invention also provides a method for preparing the compound shown in formula (I), comprising the following step A:

[0068] Step A:

[0069] Among them, X1, X2, X3, X4, X5, ring A, ring B, Y, and R a R b , m, and n have the definitions described in this article.

[0070] The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of at least one of the following: a compound of formula (I), a racemic mixture, a stereoisomer, a tautomer, a solvate, a polymorph, a pharmaceutically acceptable salt, or a prodrug compound thereof.

[0071] According to some embodiments, the pharmaceutical composition further includes one or more pharmaceutically acceptable excipients.

[0072] According to some embodiments, the pharmaceutical composition may further contain one or more additional therapeutic agents.

[0073] The present invention also provides a method for treating or preventing diseases or conditions associated with mitochondrial dysfunction, comprising administering to a patient a preventive or therapeutically effective amount of at least one of the following: a compound of formula (I), its racemic, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt, or a prodrug compound thereof, or a pharmaceutical composition thereof.

[0074] According to some implementation schemes, the diseases or conditions associated with mitochondrial dysfunction include Parkinson's disease, cancer and fibrosis, Leigh syndrome; multiple sclerosis; mitochondrial encephalopathy; mitochondrial encephalomyopathy with hyperlactatemia and stroke-like episodes (MELAS); Leber's hereditary optic neuropathy; ataxia; retinitis pigmentosa-maternally inherited Leigh syndrome; Danon's disease; diabetes; diabetic nephropathy; metabolic disorders; heart failure; ischemic heart disease leading to myocardial infarction; schizophrenia; polysulfatase deficiency; mucolipid storage disease II; mucolipid storage disease III; mucolipid storage disease IV; GML-ganglioside storage disease; neuronal ceroid lipofuscin deposition disease; Alpes disease; Barth syndrome; β-oxidation deficiency; carnitine-acyl-carnitine deficiency; carnitine deficiency; creatine deficiency syndrome; coenzyme Q10 deficiency; complex I deficiency; complex II deficiency; complex III deficiency; complex IV deficiency; complex V deficiency; COX deficiency; chronic progressive lateral oculomotor palsy syndrome; CPT. CPT II deficiency; glutaric aciduria type II; Kiehl's syndrome; lactic acidosis; long-chain acyl-CoA dehydrogenase deficiency; Lewy body disease or syndrome; fatal infantile cardiomyopathy; Left disease; medium-chain acyl-CoA dehydrogenase deficiency; myoclonic epilepsy and fluffy red fibrinoid syndrome; mitochondrial cytopathic disease; mitochondrial degenerative ataxia syndrome; mitochondrial DNA depletion syndrome; musculoskeletal disorders and encephalopathy; Pearson syndrome; pyruvate dehydrogenase deficiency; pyruvate carboxylase deficiency; POLG mutation; medium / short 3-hydroxyacyl-CoA dehydrogenase deficiency; and very long-chain acyl-CoA dehydrogenase deficiency; and age-dependent decline in cognitive function and muscle strength; amyotrophic lateral sclerosis; Huntington's disease; focal ischemia; stroke; Lewy body dementia and frontotemporal dementia.

[0075] According to some implementation schemes, the patients include mammals, preferably humans.

[0076] The present invention also provides at least one of the following: a compound of formula (I), a racemic mixture, a stereoisomer, a tautomer, a solvate, a polymorph, a pharmaceutically acceptable salt thereof, or a prodrug compound thereof, or a pharmaceutical composition thereof, for the treatment or prevention of diseases or conditions associated with mitochondrial dysfunction.

[0077] According to some implementation schemes, the diseases or conditions associated with mitochondrial dysfunction include Parkinson's disease, cancer and fibrosis, Leigh syndrome; multiple sclerosis; mitochondrial encephalopathy; mitochondrial encephalomyopathy with hyperlactatemia and stroke-like episodes (MELAS); Leber's hereditary optic neuropathy; ataxia; retinitis pigmentosa-maternally inherited Leigh syndrome; Danon's disease; diabetes; diabetic nephropathy; metabolic disorders; heart failure; ischemic heart disease leading to myocardial infarction; schizophrenia; polysulfatase deficiency; mucolipid storage disease II; mucolipid storage disease III; mucolipid storage disease IV; GML-ganglioside storage disease; neuronal ceroid lipofuscin deposition disease; Alpes disease; Barth syndrome; β-oxidation deficiency; carnitine-acyl-carnitine deficiency; carnitine deficiency; creatine deficiency syndrome; coenzyme Q10 deficiency; complex I deficiency; complex II deficiency; complex III deficiency; complex IV deficiency; complex V deficiency; COX deficiency; chronic progressive lateral oculomotor palsy syndrome; CPT. CPT II deficiency; glutaric aciduria type II; Kiehl's syndrome; lactic acidosis; long-chain acyl-CoA dehydrogenase deficiency; Lewy body disease or syndrome; fatal infantile cardiomyopathy; Left disease; medium-chain acyl-CoA dehydrogenase deficiency; myoclonic epilepsy and fluffy red fibrinoid syndrome; mitochondrial cytopathic disease; mitochondrial degenerative ataxia syndrome; mitochondrial DNA depletion syndrome; musculoskeletal disorders and encephalopathy; Pearson syndrome; pyruvate dehydrogenase deficiency; pyruvate carboxylase deficiency; POLG mutation; medium / short 3-hydroxyacyl-CoA dehydrogenase deficiency; and very long-chain acyl-CoA dehydrogenase deficiency; and age-dependent decline in cognitive function and muscle strength; amyotrophic lateral sclerosis; Huntington's disease; focal ischemia; stroke; Lewy body dementia and frontotemporal dementia.

[0078] The present invention also provides the use of at least one of the compounds of formula (I), their racemates, stereoisomers, tautomers, solvates, polymorphs, pharmaceutically acceptable salts or prodrug compounds thereof in the preparation of pharmaceuticals.

[0079] According to some implementation schemes, the use may be in the preparation of medicaments for the treatment or prevention of diseases or conditions associated with mitochondrial dysfunction.

[0080] According to some implementation schemes, the diseases or conditions associated with mitochondrial dysfunction include Parkinson's disease, cancer and fibrosis, Leigh syndrome; multiple sclerosis; mitochondrial encephalopathy; mitochondrial encephalomyopathy with hyperlactatemia and stroke-like episodes (MELAS); Leber's hereditary optic neuropathy; ataxia; retinitis pigmentosa-maternally inherited Leigh syndrome; Danon's disease; diabetes; diabetic nephropathy; metabolic disorders; heart failure; ischemic heart disease leading to myocardial infarction; schizophrenia; polysulfatase deficiency; mucolipid storage disease II; mucolipid storage disease III; mucolipid storage disease IV; GML-ganglioside storage disease; neuronal ceroid lipofuscin deposition disease; Alpes disease; Barth syndrome; β-oxidation deficiency; carnitine-acyl-carnitine deficiency; carnitine deficiency; creatine deficiency syndrome; coenzyme Q10 deficiency; complex I deficiency; complex II deficiency; complex III deficiency; complex IV deficiency; complex V deficiency; COX deficiency; chronic progressive lateral oculomotor palsy syndrome; CPT. CPT II deficiency; glutaric aciduria type II; Kiehl's syndrome; lactic acidosis; long-chain acyl-CoA dehydrogenase deficiency; Lewy body disease or syndrome; fatal infantile cardiomyopathy; Left disease; medium-chain acyl-CoA dehydrogenase deficiency; myoclonic epilepsy and fluffy red fibrinoid syndrome; mitochondrial cytopathic disease; mitochondrial degenerative ataxia syndrome; mitochondrial DNA depletion syndrome; musculoskeletal disorders and encephalopathy; Pearson syndrome; pyruvate dehydrogenase deficiency; pyruvate carboxylase deficiency; POLG mutation; medium / short 3-hydroxyacyl-CoA dehydrogenase deficiency; and very long-chain acyl-CoA dehydrogenase deficiency; and age-dependent decline in cognitive function and muscle strength; amyotrophic lateral sclerosis; Huntington's disease; focal ischemia; stroke; Lewy body dementia and frontotemporal dementia. Beneficial effects

[0081] The compounds provided by this invention can effectively inhibit the hydrolysis of ubiquitin on the outer mitochondrial membrane and accelerate mitochondrial clearance, and can be used to treat or prevent symptoms and diseases related to mitochondrial dysfunction.

[0082] Terminology Definitions and Explanations

[0083] Unless otherwise stated, the definitions of groups and terms recorded in this application specification and claims, including definitions as examples, exemplary definitions, preferred definitions, definitions recorded in tables, and definitions of specific compounds in the examples, can be arbitrarily combined and combined with each other. Such combinations and combinations of group definitions and compound structures should be understood as being within the scope of this application specification and / or claims.

[0084] The term “optional” (or “optionally”, “optionally”) in the general formula definition of this application means the case of being substituted by zero or one or more substituents. For example, “optionally substituted by one, two or more R” means that it may not be substituted by R (no substitution) or may be substituted by one, two or more R.

[0085] "More than" means three or more, such as 3, 4, 5, 6, 7, 8, 9 or 10.

[0086] Unless otherwise stated, the numerical ranges described in this specification and claims are equivalent to describing at least each specific integer value therein. For example, the numerical range "1-12" is equivalent to describing each integer value in the numerical range "1-12", namely 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12.

[0087] Term "C" 1-12 "alkyl" should be understood to refer to straight-chain and branched alkyl groups having 1 to 12 carbon atoms, "C 1-8 "Alkyl" refers to straight-chain and branched alkyl groups having 1, 2, 3, 4, 5, 6, 7, or 8 carbon atoms. 1-6 "Alkyl" means a straight-chain or branched alkyl group having 1, 2, 3, 4, 5, or 6 carbon atoms. The alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl, or 1,2-dimethylbutyl, or their isomers.

[0088] Term "C" 2-12 "Alkenyl" should be understood as representing a monovalent hydrocarbon group with 1 to 12 carbon atoms, either linear or branched, containing one or more double bonds and having 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 carbon atoms. For example, having 2, 3, 4, 5, 6, 7, or 8 carbon atoms (i.e., C...). 2-8 Alkenyl), for example, having 2, 3, 4, 5 or 6 carbon atoms (i.e., C64 ... 2-6 alkenyl), having 2 or 3 carbon atoms (i.e., C24, C34, C4 ... 2-3Alkenyl). It should be understood that when the alkenyl group contains more than one double bond, the double bonds may be separable or conjugated. The alkenyl group is, for example, vinyl, allyl, (E)-2-methylvinyl, (Z)-2-methylvinyl, (E)-but-2-enyl, (Z)-but-2-enyl, (E)-but-1-enyl, (Z)-but-1-enyl, pent-4-enyl, (E)-pent-3-enyl, (Z)-pent-3-enyl, (E)-pent-2-enyl, (Z)-pent-2-enyl, (E)- Pentyl-1-enyl, (Z)-pentyl-1-enyl, hex-5-enyl, (E)-hex-4-enyl, (Z)-hex-4-enyl, (E)-hex-3-enyl, (Z)-hex-3-enyl, (E)-hex-2-enyl, (Z)-hex-2-enyl, (E)-hex-1-enyl, (Z)-hex-1-enyl, isopropenyl, 2-methylprop-2-enyl, 1-methylprop-2-enyl 2-Methylprop-1-enyl, (E)-1-methylprop-1-enyl, (Z)-1-methylprop-1-enyl, 3-methylbut-3-enyl, 2-methylbut-3-enyl, 1-methylbut-3-enyl, 3-methylbut-2-enyl, (E)-2-methylbut-2-enyl, (Z)-2-methylbut-2-enyl, (E)-1-methylbut-2-enyl, (Z)-1-methyl But-2-enyl, (E)-3-methylbut-1-enyl, (Z)-3-methylbut-1-enyl, (E)-2-methylbut-1-enyl, (Z)-2-methylbut-1-enyl, (E)-1-methylbut-1-enyl, (Z)-1-methylbut-1-enyl, 1,1-dimethylprop-2-enyl, 1-ethylprop-1-enyl, 1-propylvinyl, 1-isopropylvinyl.

[0089] Term "C" 2-12 "Alkyne" should be understood as representing a monovalent hydrocarbon group with 1 to 12 carbon atoms, either directly linked or branched, containing one or more triple bonds and having 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 carbon atoms, for example, having 2, 3, 4, 5, 6, 7, or 8 carbon atoms (i.e., "C"). 2-8 "Alkyne group" has 2, 3, 4, 5 or 6 carbon atoms (i.e., "C"). 2-6 The alkynyl group ("C") has 2 or 3 carbon atoms ("C") 2-3The alkynyl group is, for example, ethynyl, prop-1-alkynyl, prop-2-alkynyl, but-1-alkynyl, but-2-alkynyl, but-3-alkynyl, pent-1-alkynyl, pent-2-alkynyl, pent-3-alkynyl, pent-4-alkynyl, hex-1-alkynyl, hex-2-alkynyl, hex-3-alkynyl, hex-4-alkynyl, hex-5-alkynyl, 1-methylprop-2-alkynyl, 2-methylbut-3-alkynyl, 1-methylbut-3-alkynyl, 1-methylbut-2-alkynyl, 3-methylbut-1-alkynyl, 1-ethylprop-2-alkynyl, 3-methylpent-4-alkynyl, 2-methylpent-4-alkynyl, 1-methylpent-4-alkynyl -Alynyl, 2-methylpentan-3-ynyl, 1-methylpentan-3-ynyl, 4-methylpentan-2-ynyl, 1-methylpentan-2-ynyl, 4-methylpentan-1-ynyl, 3-methylpentan-1-ynyl, 2-ethylbutan-3-ynyl, 1-ethylbutan-3-ynyl, 1-ethylbutan-2-ynyl, 1-propylpropan-2-ynyl, 1-isopropylpropan-2-ynyl, 2,2-dimethylbutan-3-ynyl, 1,1-dimethylbutan-3-ynyl, 1,1-dimethylbutan-2-ynyl, or 3,3-dimethylbutan-1-ynyl. In particular, the ynyl group is ethynyl, propan-1-ynyl, or propan-2-ynyl.

[0090] Term "C" 3-12 "Cycloalkyl" should be understood to refer to saturated monovalent monocyclic, bicyclic (such as fused ring, bridged ring, spiro ring) hydrocarbon rings or tricyclic alkanes, having 3 to 12 carbon atoms, preferably "C". 3-10 "Cycloalkyl", more preferably "C" 3-8 cycloalkyl. The term "C" 3-12 "Cycloalkyl" should be understood to refer to a saturated monovalent monocyclic, bicyclic (e.g., bridged, spirocyclic) hydrocarbon ring or tricyclic alkane having 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 carbon atoms. The C... 3-12 Cycloalkyl groups can be monocyclic hydrocarbon groups, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, or cyclodecyl; or bicyclic hydrocarbon groups, such as borneolyl, indolyl, hexahydroindolyl, tetrahydronaphthyl, decahydronaphthyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.1]heptenyl, 6,6-dimethylbicyclo[3.1.1]heptyl, 2,6,6-trimethylbicyclo[3.1.1]heptyl, bicyclo[2.2.2]octyl, 2,7-diazaspiro[3,5]nonyl, 2,6-diazaspiro[3,4]octyl; or tricyclic hydrocarbon groups, such as adamantyl.

[0091] Term "C" 6-14 "Aryl" should preferably be understood to represent a monocyclic, bicyclic (such as fused ring, bridged ring, spiro ring), or tricyclic hydrocarbon ring having 6 to 14 carbon atoms and possessing monovalent aromaticity or partial aromaticity. It can be a monoaromatic ring or a polyaromatic ring fused together, preferably "C".6-10 "Aromatic". The term "C" 6-14 "Aryl" should be understood to preferably represent a monovalent aromatic or partially aromatic monocyclic, bicyclic, or tricyclic hydrocarbon ring ("C") having 6, 7, 8, 9, 10, 11, 12, 13, or 14 carbon atoms. 6-14 Aryl), particularly a ring with 6 carbon atoms (“C6 aryl”), such as phenyl; or biphenyl, or a ring with 9 carbon atoms (“C9 aryl”), such as indenyl or indenyl, or a ring with 10 carbon atoms (“C9 aryl”). 10 Aryl groups, such as tetrahydronaphthyl, dihydronaphthyl, or naphthyl, or rings with 13 carbon atoms (“C”). 13 Aryl groups, such as fluorene groups, or rings with 14 carbon atoms (“C”). 14 Aryl), for example, anthracene. When the C 6-20 When the aryl group is substituted, it can be monosubstituted or polysubstituted. Furthermore, there are no restrictions on the substitution site; for example, it can be ortho, para, or meta substituted.

[0092] The term "5-14-membered heteroaryl" should be understood to include monocyclic, bicyclic (e.g., fused, bridged, spirocyclic), or tricyclic aromatic ring systems having 5 to 14 ring atoms and containing 1 to 5 heteroatoms independently selected from N, O, and S, for example, "5-10-membered heteroaryl". The term "5-14-membered heteroaryl" should also be understood to include monocyclic, bicyclic, or tricyclic aromatic ring systems having 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 ring atoms, particularly 5, 6, 9, or 10 carbon atoms, and containing 1 to 5, preferably 1 to 3, heteroatoms independently selected from N, O, and S, and in each case, may be benzo[a]fused. "Hyperaryl" also refers to a group in which the heteroaryl ring is fused with one or more aryl, alicyclic, or heterocyclic rings, wherein the root or point of the connection is on the heteroaryl ring. Non-limiting examples include 1-, 2-, 3-, 5-, 6-, 7- or 8-indazinyl, 1-, 3-, 4-, 5-, 6- or 7-isoindolyl, 2-, 3-, 4-, 5-, 6- or 7-indolyl, 2-, 3-, 4-, 5-, 6- or 7-indazolyl, 2-, 4-, 5-, 6-, 7- or 8-purinel, 1-, 2-, 3-, 4-, 6-, 7-, 8- or 9-quinazinyl, 2-, 3-, 4-, 5-, 6-, 7- or 8-quinolinyl, 1-, 3-, 4-... -, 5-, 6-, 7- or 8-isoquinolinyl, 1-, 4-, 5-, 6-, 7- or 8-phthalazinyl, 2-, 3-, 4-, 5- or 6-naphthidyl, 2-, 3-, 5-, 6-, 7- or 8-quinazolinyl, 3-, 4-, 5-, 6-, 7- or 8-cenolinyl, 2-, 4-, 6- or 7-pteridyl, 1-, 2-, 3-, 4-, 5-, 6-, 7- or 8-4aH carbazole, 1-, 2-, 3-, 4-, 5-, 6- 7- or 8-carbazolyl, 1-, 3-, 4-, 5-, 6-, 7-, 8- or 9-carbaolinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9- or 10-phenanthridyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, 8- or 9-acridyl, 1-, 2-, 4-, 5-, 6-, 7-, 8- or 9-pyridyl, 2-, 3-, 4-, 5-, 6-, 8-, 9- or 10-phenanthrolinel, 1-, 2-, 3-, 4-, 6-, 7-, 8- or 9- Phenazinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9- or 10-phenthiazinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9- or 10-phenazinyl, 2-, 3-, 4-, 5-, 6- or 1-, 3-, 4-, 5-, 6-, 7-, 8-, 9- or 10-benzisoquinolinyl, 2-, 3-, 4- or thieno[2,3-b]furanyl, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10- or 11-7H-pyrazin[2,3-c]carbazolyl, 2-, 3-, 5-, 6- or 7-2H-furano[3,2-b]-pyranolyl, 2-, 3-, 4-, 5-, 7- or 8-5H-pyrido[2,3-d]-o-azinyl, 1-, 3- or 5-1H-pyrazolo[4,3-d]-azolel, 2-, 4- or 5-4H-imidazo[4,5-d]thiazolyl, 3-, 5- or 8-pyrazolo[2,3-d]pyridazinyl, 2-, 3-, 5- or 6-imidazo[2,1-b]thiazolyl, 1-, 3-, 6-, 7-, 8- or 9-furano[3,4-c]cenolinyl, 1-, 2-, 3-, 4-, 5-, 6-, 8-, 9-, 10- or 11-4H-pyrido[2,3] -c]carbazolyl, 2-, 3-, 6- or 7-imidazo[1,2-b][1,2,4]triazinyl, 7-benzo[b]thiopheneyl, 2-, 4-, 5-, 6- or 7-benzozolyl, 2-, 4-, 5-, 6- or 7-benzimidazinyl, 2-, 4-, 4-, 5-, 6- or 7-benzothiazolyl, 1-, 2-, 4-, 5-, 6-, 7-, 8- or 9-benzoxapinyl, 2-, 4-, 5-, 6-, 7- or 8-benzoazinyl, 1-, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10- or 11-4H-pyrrolo[1,2-b][2]benzozapinyl. Typical fused heteroaryl groups include, but are not limited to, 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinolinyl, 1-, 3-, 4-, 5-, 6-, 7-, or 8-isoquinolinyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-, 6-, or 7-benzo[b]thiophene, 2-, 4-, 5-, 6-, or 7-benzozozolyl, 2-, 4-, 5-, 6-, or 7-benzimidazolyl, and 2-, 4-, 5-, 6-, or 7-benzothiazolyl. When the 5-14-membered heteroaryl group is linked to other groups to form the compounds of the present invention, the carbon atom on the 5-14-membered heteroaryl ring may be linked to other groups, or the heteroatom on the 5-14-membered heteroaryl ring may be linked to other groups. When the 5-14-membered heteroaryl group is substituted, it may be monosubstituted or polysubstituted. Furthermore, there are no restrictions on the substitution sites; for example, hydrogen atoms bonded to carbon atoms on the heteroaryl ring can be substituted, or hydrogen atoms bonded to heteroatoms on the heteroaryl ring can be substituted.

[0093] The term "carbocyclic ring" refers to a saturated or unsaturated non-aromatic monocyclic or polycyclic (such as bicyclic) hydrocarbon ring (e.g., monocyclic rings such as cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane, cyclononane, or bicyclic rings, including spirocyclic, fused, or bridged systems (such as bicyclic [11.1]pentane, bicyclic [2.2.1]heptane, bicyclic [3.2.1]octane, or bicyclic [5.2.0]nonane, tert-naphthalene, etc.), which may optionally be substituted with one or more (such as 1, 2, or 3) suitable substituents. The term "3-6 membered carbocyclic ring" refers to a carbocyclic ring containing 3, 4, 5, or 6 cyclic carbon atoms.

[0094] Unless otherwise defined, the term "3-14 membered heterocyclic group" refers to a saturated or unsaturated non-aromatic ring or ring system, for example, a 4-, 5-, 6-, or 7-membered monocyclic ring, a 7-, 8-, 9-, 10-, 11-, or 12-membered bicyclic ring (such as a fused ring, bridged ring, or spirocyclic ring), or a 10-, 11-, 12-, 13-, or 14-membered tricyclic ring system, and contains at least one, for example, 1, 2, 3, 4, 5, or more heteroatoms selected from O, S, and N, wherein N and S may optionally be oxidized to various oxidation states to form nitrogen oxides, -S(O)-, or -S(O)2- states. For example, the "3-14 membered heterocyclic group" may be a 3-14 membered N-containing heterocyclic group (containing at least one N). Preferably, the heterocyclic group may be selected from "3-10 membered heterocyclic groups". The term "3-10 membered heterocyclic group" refers to a saturated or unsaturated non-aromatic ring or ring system containing at least one heteroatom selected from O, S, and N. The heterocyclic group can be connected to the remainder of the molecule via any one of the carbon atoms or a nitrogen atom (if present). The heterocyclic group can include fused or bridged rings and spirocyclic rings. Specifically, the heterocyclic group can include, but is not limited to: 4-membered rings, such as azirrobutyl or oxobutyl; 5-membered rings, such as tetrahydrofuranyl, dioxacyclopentenyl, pyrrolyl, imidazoalkyl, pyrazolyl, or pyrrololinyl; or 6-membered rings, such as tetrahydropyranyl, piperidinyl, morpholinyl, dithiaalkyl, thiomorpholinyl, piperazinyl, or trithiaalkyl; or 7-membered rings, such as diazacycloheptyl. Optionally, the heterocyclic group can be benzofused. The heterocyclic group can be bicyclic, such as, but not limited to, a 5,5-membered ring, like a hexahydrocyclopentano[c]pyrrole-2(1H)-yl ring, or a 5,6-membered bicyclic ring, like a hexahydropyrrolo[1,2-a]pyrazin-2(1H)-yl ring. The heterocyclic group can be partially unsaturated, meaning it can contain one or more double bonds, such as, but not limited to, dihydrofuranyl, dihydropyranyl, 2,5-dihydro-1H-pyrroleyl, 4H-[1,3,4]thiadiazinyl, 1,2,3,5-tetrahydrooxazolyl, or 4H-[1,4]thiazinyl, or it can be benzofused, such as, but not limited to, dihydroisoquinolinyl. When the 3-14-membered heterocyclic group is linked to other groups to form the compounds of the present invention, the carbon atom on the 3-14-membered heterocyclic group can be linked to other groups, or the heterocyclic atom on the 3-14-membered heterocyclic ring can be linked to other groups. For example, when the 3-14 membered heterocyclic group is selected from piperazine, the nitrogen atom on the piperazine group can be attached to other groups. Or when the 3-14 membered heterocyclic group is selected from piperidinium, the nitrogen atom on the piperidinium ring and the carbon atom at its para position can be attached to other groups.

[0095] The term "halogen" refers to fluorine, chlorine, bromine, and iodine.

[0096] The term "nitrogen oxides" refers to compounds formed by the oxidation of nitrogen atoms in the structure of tertiary amines or nitrogen-containing (aromatic) heterocyclic compounds.

[0097] The term "spirocycle" refers to a ring system in which two rings share a single ring atom.

[0098] The term "fused ring" refers to a ring system in which two rings share two cyclic atoms.

[0099] The term "bridged ring" refers to a ring system in which two rings share three or more cyclic atoms.

[0100] Unless otherwise stated, heterocyclic, hypocyclic, heteroaryl, or hypoaryl includes all possible isomers, such as their positional isomers. Thus, for some illustrative, non-limiting examples, it may include forms in which one, two, or more of the following positions (if present) are substituted or bonded to other groups, including pyridin-2-yl, pyridin-2-yl, pyridin-3-yl, pyridin-3-yl, pyridin-4-yl, and pyridin-4-yl; thiophene or thiophene includes thiophene-2-yl, thiophene-2-yl, thiophene-3-yl, and thiophene-3-yl; pyrazol-1-yl, pyrazol-3-yl, pyrazol-4-yl, and pyrazol-5-yl.

[0101] The compounds disclosed herein may exist in various tautomer forms, and all such forms are included within the scope of this disclosure. The terms "tautomer" or "tautomer form" refer to a structural isomer that exists in equilibrium and readily transforms from one isomer to another. This includes all possible tautomers, i.e., existing as a single isomer or as a mixture of said tautomers in any proportion. Non-limiting examples include: keto-enols, imine-enamines, lactam-lactamimides, etc.

[0102] "Halogenation" refers to the replacement of a substance by one or more halogens.

[0103] The term "halogenated alkyl" refers to an alkyl group that has been substituted with one or more halogens, wherein the alkyl group is as defined above.

[0104] The term "oxo" refers to the substitution of a carbon, nitrogen, or sulfur atom in a substituent with an oxygen atom (=O) formed by oxidation.

[0105] The term "alkylamino" refers to -NH-(alkyl) or -N-(alkyl)2, where alkyl is defined as described above. Non-limiting examples of alkylamino include: methylamino, ethylamino, propylamino, isopropylamino, butylamino, dimethylamino, methylethylamino, diethylamino, dipropylamino, methylpropylamino, diisopropylamino, dibutylamino, etc.

[0106] "Hydroxyalkyl" refers to an alkyl group substituted with one or more hydroxyl groups, wherein the alkyl group is as defined above. Non-limiting examples of hydroxyalkyl groups include: hydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxymethylpropyl, or dihydroxypropyl, etc.

[0107] The term "alkyloxy" refers to -O-(alkyl), where alkyl is defined as described above. Non-limiting examples of alkoxy groups include: methoxy, ethoxy, propoxy, and butoxy. Alkoxy groups can be optionally substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkyloxy, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, or heterocycloalkyloxy.

[0108] The terms "alkyleneoxy" and "oxyalkylene" refer to -alkylene-O- or -O-alkylene-, where alkylene represents a straight-chain or branched saturated divalent hydrocarbon group. The definition of the number of carbon atoms in "alkylene" follows the definition of "alkyl" above. Those skilled in the art will understand that alkyleneoxy or oxyalkylene can be attached to the remainder of the molecule containing it in any orientation; that is, the two are used interchangeably.

[0109] Wavy lines intersecting chemical bonds Used to indicate the connection position of a group to other atoms in the molecular structure. For example... This indicates the 3-position connection with the pyridinium group. When the group connection position is not fixed, taking the pyridinium group as an example, it can be represented as follows: The method is shown to indicate that it can be connected to any connectable site on the pyridinyl group. For example... This indicates that it can be connected to any connectable position on the heteroaryl ring, for example, it can be connected to any of the four carbon atoms on the pyridine ring to the right of the heteroaryl group, or it can be connected to a carbon atom on the pyrazole ring to the left. Unless otherwise stated, similar expressions in this application are interpreted in the same way as above.

[0110] In the chemical structure of the compound described in this invention, the bond... This indicates that no configuration has been specified. Indicates absolute configuration, that is, if stereoisomers exist in the chemical structure, the bonds... It can be Or simultaneously include Two configurations.

[0111] In this invention, the compounds involved also include isotopically labeled compounds, which are the same as those shown in Formula I, but in which one or more atoms are replaced by atoms with atomic masses or mass numbers different from those normally found in nature. Examples of isotopes that can be incorporated into the compounds of this invention include isotopes of H, C, N, O, S, F, and Cl, respectively such as 2 H, 3 H, 13 C 11 C 14 C 15 N、 18 O、 17 O、 32 P, 35 S, 18 F and 36 Cl. Compounds of the present invention, their prodrugs, or pharmaceutically acceptable salts of said compounds or prodrugs containing the aforementioned isotopes and / or other isotopes are within the scope of the present invention. Certain isotopically labeled compounds of the present invention, such as those doped with radioactive isotopes (e.g.,... 3 H and 14 Compounds in (C) can be used for drug and / or substrate tissue distribution assays. Tritium (i.e., 3 H) and carbon-14 (i.e. 14 C) Isotopes are particularly preferred due to their ease of preparation and detectability. Furthermore, heavier isotopes (such as deuterium, i.e., 2 H or D substitutions can provide certain therapeutic advantages derived from greater metabolic stability (e.g., increased in vivo half-life or reduced dose requirements) and are therefore preferred in some cases. The presence of hydrogen in the substituents of this invention, without the separate mention of the terms deuterium or tritium, does not imply the exclusion of deuterium or tritium, but rather may also include deuterium or tritium.

[0112] Those skilled in the art will understand that the compounds shown in formula (I) can exist in the form of various pharmaceutically acceptable salts. If these compounds have a basic center, they can form acid addition salts; if these compounds have an acidic center, they can form base addition salts; if these compounds contain both an acidic center (e.g., a carboxyl group) and a basic center (e.g., an amino group), they can also form internal salts.

[0113] The compounds of the present invention may exist as solvates (such as hydrates), wherein the compounds of the present invention contain a polar solvent, particularly, for example, water, methanol, or ethanol, as a structural element of the lattice of the compound. The amount of the polar solvent, particularly water, may be stoichiometric or non-stoichiometric.

[0114] Depending on their molecular structure, the compounds of the present invention can be chiral, and therefore may exist in various enantiomeric forms. Thus, these compounds can exist in racemic or optically active forms. The compounds of the present invention encompass isomers of each chiral carbon in the R or S configuration, or mixtures thereof, and racemates. The compounds of the present invention or their intermediates can be isolated as enantiomeric compounds by chemical or physical methods known to those skilled in the art, or used in this form for synthesis. In the case of racemic amines, diastereomers are obtained from the mixture by reaction with an optically active resolving agent. Examples of suitable resolving agents are optically active acids, such as tartaric acid in both R and S forms, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, suitable N-protected amino acids (e.g., N-benzoylproline or N-benzenesulfonylproline), or various optically active camphorsulfonic acids. Chromatographic enantiomeric separation can also be advantageously performed using optically active resolving agents (e.g., dinitrobenzoylphenylglycine immobilized on silica gel, cellulose triacetate or other carbohydrate derivatives, or chiral derivatized isobutylene ester polymers). Suitable eluents for this purpose are aqueous or alcoholic solvent mixtures, such as hexane / isopropanol / acetonitrile.

[0115] The corresponding stable isomers can be separated using known methods, such as extraction, filtration, or column chromatography.

[0116] The term "patient" refers to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses, or primates, with humans being the most preferred.

[0117] The term “therapeutic effective amount” refers to the amount of an active compound or drug that researchers, veterinarians, physicians, or other clinicians are searching for in tissues, systems, animals, individuals, or humans to elicit a biological or medical response. It includes one or more of the following: (1) prevention of disease: e.g., prevention of disease, disorder, or condition in individuals susceptible to disease, disorder, or symptom but not yet experiencing or exhibiting the pathology or symptoms of the disease; (2) suppression of disease: e.g., suppression of disease, disorder, or symptom in individuals experiencing or exhibiting the pathology or symptoms of the disease, disorder, or symptom (i.e., prevention of further development of the pathology and / or symptoms); (3) relief of disease: e.g., relief of disease, disorder, or symptom in individuals experiencing or exhibiting the pathology or symptoms of the disease, disorder, or symptom (i.e., reversal of the pathology and / or symptoms). Detailed Implementation

[0118] The technical solution of the present invention will be further described in detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanatory of the present invention, and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection intended by the present invention.

[0119] Unless otherwise stated, the raw materials and reagents used in the following examples are commercially available products or can be prepared by known methods.

[0120] The structures of the compounds of this invention were determined by nuclear magnetic resonance (NMR) and / or liquid chromatography-mass spectrometry (LC-MS). NMR chemical shifts (δ) are given in parts per million (ppm). NMR measurements were performed using a Bruker AVANCE-400 NMR spectrometer with deuterated dimethyl sulfoxide (DMSO-d6), deuterated methanol (CD3OD), or deuterated chloroform (CDCl3) as the solvent, and tetramethylsilane (TMS) as the internal standard.

[0121] LC-MS was performed using an Agilent 1200 Infinity Series mass spectrometer. HPLC was performed using an Agilent 1200DAD high-performance liquid chromatograph (Sunfire C18 150×4.6 mm column) and a Waters 2695-2996 high-performance liquid chromatograph (Gimini C18 150×4.6 mm column).

[0122] Thin-layer chromatography (TLC) uses Yantai Huanghai HSGF254 or Qingdao Haiyang Chemical GF254 silica gel plates. The standard size for TLC is 0.15mm to 0.20mm, while the standard size for TLC separation and purification is 0.4mm to 0.5mm. Column chromatography generally uses Yantai Huanghai 200-300 mesh silica gel as the carrier.

[0123] Unless otherwise stated, the raw materials and reagents used in the following examples are commercially available or can be prepared by known methods. Unless otherwise specified, all reactions of the present invention are carried out under continuous magnetic stirring, in a dry nitrogen or argon atmosphere, using a dry solvent, and the reaction temperature is in degrees Celsius.

[0124] Example 1 (R)-N-(1-cyanopyrrolidone-3-yl)-7-(4-fluorophenyl)benzo[d]oxazol-2-carboxamide (compound Cpd-01)

[0125] Step 1: Preparation of 3-amino-4-fluoro-[1,1-biphenyl]-2-ol (Cpd-01a)

[0126] 2-Amino-6-bromophenol (900 mg, 4.8 mmol), (4-fluorophenyl)boric acid (803.7 mg, 5.7 mmol), potassium carbonate (1.32 g, 9.6 mmol), and [1,1'-bis(di-tert-butylphosphine)ferrocene]palladium dichloride (347.3 mg, 0.48 mmol) were dissolved in 1,4-dioxane / water (20 mL / 5 mL) and stirred at 90 °C for 16 h. After the reaction was completed, the solution was concentrated under reduced pressure and purified by column chromatography (ethyl acetate / petroleum ether = 15%) to give 3-amino-4-fluoro-[1,1-biphenyl]-2-ol (Cpd-01a, 300 mg), yield: 31%.

[0127] MS m / z (ESI): 204.2 (M+1)

[0128] Step 2: Preparation of ethyl 7-(4-fluorophenyl)benzo[d]oxazol-2-carboxylate (Cpd-01b)

[0129] 3-Amino-4-fluoro-[1,1-biphenyl]-2-ol (Cpd-01a, 290 mg, 1.43 mmol) and triethylamine (288.82 mg, 2.85 mmol) were dissolved in ultra-dry tetrahydrofuran (5 mL). Oxaloyl chloride monoethyl ester (214.33 mg, 1.57 mmol) was added under nitrogen protection at 0 °C. The reaction mixture was stirred at room temperature for 3 h. Then, triphenylphosphine (804.78 mg, 3.07 mmol) and diisopropyl azodicarbonate (317.43 mg, 1.57 mmol) were added sequentially, and the mixture was stirred at room temperature for 16 h. After the reaction was complete, the product was concentrated under reduced pressure and purified by column chromatography (ethyl acetate / petroleum ether = 32%) to give ethyl 7-(4-fluorophenyl)benzo[d]oxazol-2-carboxylate (Cpd-01b, 90 mg), yield: 22%.

[0130] MS m / z (ESI): 286.1 (M+1)

[0131] Step 3: Preparation of (R)-3-(7-(4-fluorophenyl)benzo[d]oxazol-2-carboxamido)pyrrolidine-1-carboxylic acid tert-butyl ester (Cpd-01c)

[0132] Ethyl 7-(4-fluorophenyl)benzo[d]oxazol-2-carboxylate (Cpd-01b, 90 mg, 0.32 mmol) was dissolved in tetrahydrofuran (2 mL). Tert-butyl (R)-3-aminopyrrolidine-1-carboxylate (88 mg, 0.47 mmol) and 1,8-diazobisspirocyclic [5.4.0]undecyl-7-ene (480 mg, 3.16 mmol) were added to the reaction solution, and the mixture was stirred at room temperature for 16 h. After the reaction was complete, water was added to the reaction solution, and the mixture was extracted with ethyl acetate, dried over anhydrous sodium sulfate, and the organic phase was concentrated under reduced pressure and purified by column chromatography (ethyl acetate / petroleum ether = 36%) to give (R)-3-(7-(4-fluorophenyl)benzo[d]oxazol-2-carboxamido)pyrrolidine-1-carboxylate (Cpd-01c, 80 mg), yield: 59%.

[0133] MS m / z (ESI): 426.1 (M+1)

[0134] Step 4: Preparation of (R)-7-(4-fluorophenyl)-N-(pyrrolidine-3-yl)benzo[d]oxazol-2-carboxamide trifluoroacetate (Cpd-01d)

[0135] (R)-3-(7-(4-fluorophenyl)benzo[d]oxazol-2-carboxamido)pyrrolidine-1-carboxylic acid tert-butyl ester (Cpd-01c, 80 mg, 0.19 mmol) was dissolved in dichloromethane (1 mL), and then trifluoroacetic acid (1 mL) was added. The reaction mixture was stirred at room temperature for 1 h. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to obtain crude (R)-7-(4-fluorophenyl)-N-(pyrrolidine-3-yl)benzo[d]oxazol-2-carboxamide trifluoroacetate (Cpd-01d, 60 mg), yield: 98%.

[0136] MS m / z (ESI): 326.1 (M+1)

[0137] Step 5: Preparation of (R)-N-(1-cyanopyrrolidone-3-yl)-7-(4-fluorophenyl)benzo[d]oxazol-2-carboxamide (Cpd-01)

[0138] (R)-7-(4-fluorophenyl)-N-(pyrrolidine-3-yl)benzo[d]oxazol-2-carboxamide trifluoroacetate (Cpd-01d, 60 mg, 0.18 mmol), cyanogen bromide (21 mg, 0.20 mmol), and potassium carbonate (76 mg, 0.55 mmol) were dissolved in tetrahydrofuran (2 mL), and stirred at room temperature for 1 h. After the reaction was complete, the organic phase was concentrated, and the crude product was purified by column chromatography (ethyl acetate / petroleum ether = 25%) to give (R)-N-(1-cyanopyrrolidine-3-yl)-7-(4-fluorophenyl)benzo[d]oxazol-2-carboxamide (Cpd-01, 48 mg), yield: 74%.

[0139] MS m / z (ESI): 351.1 (M+1)

[0140] HPLC: 96.61% (214nm), 97.89% (254nm).

[0141] 1 H NMR (400MHz, DMSO) δ9.64(d,J=6.8Hz,1H),8.02–7.92(m,2H),7.89(d,J=8.0Hz,1H),7.81(d,J=7.6Hz,1H),7.60(t,J=7 .6Hz,1H),7.43(t,J=8.8Hz,2H),4.53(dd,J=12.0,5.6Hz,1H),3.69–3.55(m,2H),3.49–3.39(m,2H),2.19–2.02(m,2H).

[0142] Example 2: N-(3-cyano-3-azabicyclo[3.1.0]hexane-1-yl)-7-(4-fluorophenyl)benzo[d]oxazol-2-carboxamide (compound Cpd-02)

[0143] Step 1: Preparation of (R)-7-(7-(4-fluorophenyl)benzo[d]oxazol-2-carboxamido)-5-azaspiro[2,4]heptane-5-carboxylic acid tert-butyl ester (Cpd-02a)

[0144] Ethyl 7-(4-fluorophenyl)benzo[d]oxazol-2-carboxylate (Cpd-01b, 50 mg, 0.175 mmol) was dissolved in 1 mL of tetrahydrofuran, followed by the addition of 1,8-diazabicycloundec-7-ene (53 mg, 0.53 mmol) and (R)-7-amino-5-azaspiro[2,4]heptane-5-carboxylate tert-butyl ester (44 mg, 0.21 mmol). The reaction mixture was stirred at 20 °C for 16 h. After the reaction was completed, the reaction mixture was concentrated and purified by silica gel agar plate preparation (petroleum ether / ethyl acetate = 1 / 1) to obtain the product (R)-7-(7-(4-fluorophenyl)benzo[d]oxazol-2-carboxamido)-5-azaspiro[2,4]heptane-5-carboxylate tert-butyl ester (Cpd-02a, 30 mg), yield: 34.11%.

[0145] MS m / z (ESI): 396.1 (M-55)

[0146] Step 2: Preparation of (R)-7-(4-fluorophenyl)-N-(5-azaspiro[2,4]heptane-7-yl)benzo[d]oxazol-2-carboxamide trifluoroacetate (Cpd-02b)

[0147] (R)-7-(7-(4-fluorophenyl)benzo[d]oxazol-2-carboxamido)-5-azaspiro[2.4]heptane-5-carboxylic acid tert-butyl ester (Cpd-02a, 30 mg, 0.068 mmol) was dissolved in 1 mL of dichloromethane, followed by the addition of 1 mL of trifluoroacetic acid. The reaction mixture was stirred at 20 °C for 2 h. After the reaction was completed, the reaction mixture was concentrated to obtain crude (R)-7-(4-fluorophenyl)-N-(5-azaspiro[2.4]heptane-7-yl)benzo[d]oxazol-2-carboxamide trifluoroacetate (Cpd-02b, 20 mg), yield: 77.84%.

[0148] MS m / z (ESI): 352.1 (M+1)

[0149] Step 3: Preparation of N-(3-cyano-3-azabicyclo[3.1.0]hexane-1-yl)-7-(4-fluorophenyl)benzo[d]oxazol-2-carboxamide (compound Cpd-02)

[0150] (R)-7-(4-fluorophenyl)-N-(5-azaspiro[2.4]heptane-7-yl)benzo[d]oxazol-2-carboxamide trifluoroacetate (Cpd-02b, 20 mg, 0.057 mmol) was dissolved in dichloromethane, followed by the addition of N,N-diisopropylethylamine (17 mg, 0.129 mmol), and then cyanogen bromide (8 mg, 0.077 mmol). The reaction mixture was stirred at 20 °C for 1 h. After the reaction was completed, the reaction solution was concentrated and then purified by silica gel stencil preparation (petroleum ether / ethyl acetate = 1 / 1) to obtain the product (R)-6-(1-(3,5-dichloropyridin-4-yl)ethoxy)-1-(6-(methylthio)pyridin-3-yl)-1,3-dihydro-2H-imidazo[4,5-b]pyridin-2-one (Cpd-02, 4.16 mg), yield: 12.88%.

[0151] MS m / z (ESI): 377.0 (M+1)

[0152] HPLC:98.29%(214nm),99.70%(254nm)

[0153] 1 H NMR(400MHz,DMSO)δ9.72(d,J=8.0Hz,1H),8.01–7.93(m,2H),7.90(d,J=8.0Hz,1H) ,7.81(d,J=7.6Hz,1H),7.61(t,J=7.6Hz,1H),7.43(t,J=8.8Hz,2H),4.31–4.20(m, 1H),3.88(dd,J=10.0,6.5Hz,1H),3.76(d,J=8.8Hz,1H),3.58(dd,J=10.0,3.4Hz,1 H),3.23(d,J=8.8Hz,1H),0.90–0.81(m,1H),0.80–0.68(m,2H),0.67–0.60(m,1H).

[0154] Example 3: N-(3-cyano-3-azabicyclo[3.1.0]hexane-1-yl)-7-(4-fluorophenyl)benzo[d]oxazol-2-carboxamide (compound Cpd-05)

[0155] Step 1: Preparation of 1-(7-(4-fluorophenyl)benzo[d]oxazol-2-carboxamido)-3-azabicyclo[3.1.0]hexane-3-carboxylic acid tert-butyl ester (Cpd-05a)

[0156] Ethyl 7-(4-fluorophenyl)benzo[d]oxazol-2-carboxylate (Cpd-01b, 50 mg, 0.175 mmol) was dissolved in 1 mL of tetrahydrofuran, followed by the addition of 1,8-diazabicycloundec-7-ene (53 mg, 0.53 mmol) and 1-amino-3-azabicyclo[3.1.0]hexane-3-carboxylate tert-butyl ester (41 mg, 0.21 mmol). The reaction mixture was stirred at 20 °C for 16 h. After the reaction was completed, the reaction mixture was concentrated and purified by silica gel agar plate preparation (petroleum ether / ethyl acetate = 1 / 1) to obtain the product 1-(7-(4-fluorophenyl)benzo[d]oxazol-2-carboxamido)-3-azabicyclo[3.1.0]hexane-3-carboxylate tert-butyl ester (Cpd-05a, 30 mg), yield: 35%.

[0157] MS m / z (ESI): 382.1 (M-55)

[0158] Step 2: Preparation of N-(3-azabicyclo[3.1.0]hexane-1-yl)-7-(4-fluorophenyl)benzo[d]oxazol-2-carboxamide trifluoroacetate (Cpd-05b)

[0159] 1-(7-(4-fluorophenyl)benzo[d]oxazol-2-carboxamido)-3-azabicyclo[3.1.0]hexane-3-carboxylic acid tert-butyl ester (Cpd-05a, 30 mg, 0.068 mmol) was dissolved in 1 mL of dichloromethane, followed by the addition of 1 mL of trifluoroacetic acid. The reaction mixture was stirred at 20 °C for 2 h. After the reaction was completed, the reaction mixture was concentrated to obtain crude N-(3-azabicyclo[3.1.0]hexane-1-yl)-7-(4-fluorophenyl)benzo[d]oxazol-2-carboxamide trifluoroacetate (Cpd-05b, 20 mg), yield: 77.84%.

[0160] MS m / z (ESI): 338.1 (M+1)

[0161] Step 3: Preparation of N-(3-cyano-3-azabicyclo[3.1.0]hexane-1-yl)-7-(4-fluorophenyl)benzo[d]oxazol-2-carboxamide (Cpd-05)

[0162] N-(3-azabicyclo[3.1.0]hexan-1-yl)-7-(4-fluorophenyl)benzo[d]oxazol-2-carboxamide trifluoroacetate (Cpd-05b, 20 mg, 0.059 mmol) was dissolved in dichloromethane, followed by the addition of N,N-diisopropylethylamine (18 mg, 0.134 mmol), and then cyanogen bromide (9 mg, 0.079 mmol). The reaction mixture was stirred at 20 °C for 1 h. After the reaction was completed, the reaction mixture was concentrated and purified by silica gel agar (petroleum ether / ethyl acetate = 1 / 1) to obtain the product N-(3-cyano-3-azabicyclo[3.1.0]hexan-1-yl)-7-(4-fluorophenyl)benzo[d]oxazol-2-carboxamide (Cpd-05, 3.52 mg), yield: 10.91%.

[0163] MS m / z (ESI): 363.0 (M+1)

[0164] HPLC:100%(214nm),99.62%(254nm)

[0165] 1 H NMR(400MHz,DMSO)δ9.86(s,1H),8.05–7.93(m,2H),7.88(d,J=8.0Hz,1H),7.81(d,J =6.8Hz,1H),7.59(t,J=7.6Hz,1H),7.43(t,J=8.8Hz,2H),3.71(dd,J=9.6,3.9Hz,1H ), 3.60 (dd, J = 21.8, 9.2 Hz, 3H), 3.39 (d, J = 9.7 Hz, 1H), 1.93–1.85 (m, 1H), 0.94 (t, J = 5.6 Hz, 1H). Example 4 N-(2-cyano-2-azaspiro[3.3]hept-6-yl)-7-(4-fluorophenyl)benzo[d]oxazol-2-carboxamide (compound Cpd-24)

[0166] Step 1: Preparation of tert-butyl 6-(7-(4-fluorophenyl)benzo[d]oxazol-2-carbamate)-2-azaspiro[3.3]heptane-2-carboxylate (Cpd-24a)

[0167] Ethyl 7-(4-fluorophenyl)benzo[d]oxazol-2-carboxylate (Cpd-01b, 50 mg, 0.175 mmol) and tert-butyl 6-amino-2-azaspiro[3.3]heptane-2-carboxylate (37 mg, 0.175 mmol) were dissolved in tetrahydrofuran (0.6 mL), and 1,8-diazabicyclo[5.4.0]undec-7-ene (214 mg, 1.40 mmol) was added. The reaction mixture was stirred for 16 h at room temperature under nitrogen protection. After the reaction was completed, the reaction mixture was poured into water and extracted with ethyl acetate (5 mL × 2). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated to dryness, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to give tert-butyl 6-(7-(4-fluorophenyl)benzo[d]oxazol-2-carboxamido)-2-azaspiro[3.3]heptane-2-carboxylate (Cpd-24a, 60 mg), yield: 75%.

[0168] MS m / z (ESI): 396.1 (M+1-56)

[0169] Step 2: Preparation of (7-(4-fluorophenyl)-N-2-azaspiro[3.3]hept-6-yl)benzo[d]oxazol-2-carboxamide trifluoroacetate (Cpd-24b)

[0170] 6-(7-(4-fluorophenyl)benzo[d]oxazol-2-carboxamido)-2-azaspiro[3.3]heptane-2-carboxylic acid tert-butyl ester (Cpd-24a, 60 mg, 0.133 mmol) was dissolved in dichloromethane (1 mL), and trifluoroacetic acid (303 mg, 2.66 mmol) was added. The reaction mixture was stirred at 25 °C for 1 h. After the reaction was completed, the reaction mixture was concentrated to dryness to obtain (7-(4-fluorophenyl)-N-2-azaspiro[3.3]hept-6-yl)benzo[d]oxazol-2-carboxamide trifluoroacetate (Cpd-24b, 60 mg), yield: 96%.

[0171] MS m / z (ESI): 352.2 (M+1)

[0172] Step 3: Preparation of N-(2-cyano-2-azaspiro[3.3]hept-6-yl)-7-(4-fluorophenyl)benzo[d]oxazol-2-carboxamide (Cpd-24)

[0173] (7-(4-fluorophenyl)-N-2-azaspiro[3.3]hept-6-yl)benzo[d]oxazol-2-carboxamide trifluoroacetate (Cpd-24b, 60 mg, 0.129 mmol) was dissolved in dichloromethane (2 mL), and diisopropylethylamine (100 mg, 0.773 mmol) and cyanogen bromide (15 mg, 0.142 mmol) were added. The reaction mixture was stirred at 25 °C for 1 h. After the reaction was complete, water was added to the reaction mixture to adjust the pH to 5, and the mixture was extracted with dichloromethane, washed with brine, and concentrated to dryness. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to give N-(2-cyano-2-azaspiro[3.3]hept-6-yl)-7-(4-fluorophenyl)benzo[d]oxazol-2-carboxamide (Cpd-24, 32.73 mg), yield: 67%.

[0174] MS m / z (ESI): 377.1 (M+1)

[0175] HPLC: 95.53% (214nm), 98.80% (254nm).

[0176] 1 H NMR (400MHz, DMSO) δ9.55(d,J=7.8Hz,1H),7.99–7.93(m,2H),7.87(d,J=7.2Hz,1H),7.79(d,J=7.0Hz,1H),7.59(t,J=7.8H z,1H),7.43(t,J=9.0Hz,2H),4.28(dd,J=16.2,8.2Hz,1H),4.22(s,2H),4.11(s,2H),2.60–2.54(m,2H),2.42–2.35(m,2H).

[0177] Example 5 7-(7-(4-fluorophenyl)benzo[d]oxazol-2-carbonyl)-2,7-diazaspiro[4.4]nonane-2-nitrile (compound Cpd-26)

[0178] Step 1: Preparation of tert-butyl 7-(7-(4-fluorophenyl)benzo[d]oxazol-2-carbonyl)-2,7-diazaspiro[4.4]nonane-2-carboxylate (Cpd-26a)

[0179] Ethyl 7-(4-fluorophenyl)benzo[d]oxazol-2-carboxylate (Cpd-01b, 50 mg, 0.175 mmol) and tert-butyl 2,7-diazaspiro[4.4]nonane-2-carboxylate (40 mg, 0.175 mmol) were dissolved in tetrahydrofuran (0.6 mL), and 1,8-diazabicyclo[5.4.0]undec-7-ene (214 mg, 1.40 mmol) was added. The reaction mixture was stirred for 16 h at room temperature under nitrogen protection. After the reaction was completed, the reaction mixture was poured into water and extracted with ethyl acetate (5 mL × 2). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated to dryness, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to give tert-butyl 7-(7-(4-fluorophenyl)benzo[d]oxazol-2-carbonyl)-2,7-diazaspiro[4.4]nonane-2-carboxylate (Cpd-26a, 60 mg), yield: 73%.

[0180] MS m / z (ESI): 410.1 (M+1-56)

[0181] Step 2: Preparation of (7-(4-fluorophenyl)benzo[d]oxazol-2-yl)(2,7-diazaspiro[4.4]non-2-yl)methyl ketone trifluoroacetate (Cpd-26b)

[0182] 7-(7-(4-fluorophenyl)benzo[d]oxazol-2-carbonyl)-2,7-diazaspiro[4.4]nonane-2-carboxylic acid tert-butyl ester (Cpd-26a, 60 mg, 0.129 mmol) was dissolved in dichloromethane (1 mL), and trifluoroacetic acid (294 mg, 2.58 mmol) was added. The reaction mixture was stirred at 25 °C for 1 h. After the reaction was completed, the reaction mixture was concentrated to dryness to obtain (7-(4-fluorophenyl)benzo[d]oxazol-2-yl)(2,7-diazaspiro[4.4]non-2-yl)methyl ketone trifluoroacetate (Cpd-26b, 60 mg), yield: 97%.

[0183] MS m / z (ESI): 366.1 (M+1)

[0184] Step 3: Preparation of 7-(7-(4-fluorophenyl)benzo[d]oxazol-2-carbonyl)-2,7-diazaspiro[4.4]nonane-2-nitrile (Cpd-26)

[0185] (7-(4-fluorophenyl)benzo[d]oxazol-2-yl)(2,7-diazaspiro[4.4]non-2-yl)methyl ketone trifluoroacetate (Cpd-26b, 60 mg, 0.125 mmol) was dissolved in dichloromethane (2 mL), and diisopropylethylamine (97 mg, 0.751 mmol) and cyanogen bromide (15 mg, 0.138 mmol) were added. The reaction mixture was stirred at 25 °C for 1 h. After the reaction was complete, water was added to the reaction mixture to adjust the pH to 5, and the mixture was extracted with dichloromethane, washed with brine, and concentrated to dryness. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 5:1) to give 7-(7-(4-fluorophenyl)benzo[d]oxazol-2-carbonyl)-2,7-diazaspiro[4.4]nonane-2-onitrile (Cpd-26, 33.80 mg), yield: 69%.

[0186] MS m / z (ESI): 391.1 (M+1)

[0187] HPLC: 100% (214nm), 100% (254nm).

[0188] 1 H NMR (400MHz, DMSO) δ8.02–7.94(m,2H),7.91(t,J=7.6Hz,1H),7.80(d,J=7.6 Hz,1H),7.59(t,J=8.0Hz,1H),7.43(t,J=9.0Hz,2H),4.19–4.07(m,1H),3.99 (dd,J=27.2,11.6Hz,1H),3.69–3.64(m,1H),3.58(d,J=20.6Hz,1H),3.52–3 .46(m,2H),3.43(d,J=9.4Hz,1H),3.36(d,J=10.6Hz,1H),2.02–1.85(m,4H).

[0189] Example 6 (R)-N-(5-cyano-5-azaspiro[2.4]heptane-7-yl)-5-(2-cyanopyridin-4-yl)oxazol-2-carboxamide (compound Cpd-35A)

[0190] The intermediate ethyl 5-(2-cyanopyridin-4-yl)-1,3-oxazol-2-carboxylate (Cpd-35A-a) was synthesized according to patent WO2021249909.

[0191] Step 1: Preparation of (R)-7-(5-(2-cyanopyridin-4-yl)oxazol-2-carboxamido)-5-azaspiro[2.4]heptane-5-carboxylic acid tert-butyl ester (Cpd-35A-b)

[0192] Ethyl 5-(2-cyanopyridin-4-yl)-1,3-oxazol-2-carboxylate (Cpd-35A-a, 30 mg, 0.123 mmol) was dissolved in 1 mL of tetrahydrofuran, followed by the addition of 1,8-diazabicycloundec-7-ene (56 mg, 0.36 mmol) and tert-butyl 1-amino-3-azabicyclo[3.1.0]hexane-3-carboxylate (28 mg, 0.135 mmol). The reaction mixture was stirred at 20 °C for 16 h. After the reaction was completed, the reaction mixture was concentrated and purified by silica gel ablation (petroleum ether / ethyl acetate = 1 / 1) to obtain product (R)-7-(5-(2-cyanopyridin-4-yl)oxazol-2-carboxamido)-5-azaspiro[2.4]heptane-5-carboxylate tert-butyl ester (Cpd-35A-b, 30 mg), yield: 53%.

[0193] MS m / z (ESI): 354.0 (M-55)

[0194] Step 2: Preparation of (R)-5-(2-cyanopyridin-4-yl)-N-(5-azaspiro[2.4]heptane-7-yl)oxazol-2-carboxamide trifluoroacetate (Cpd-35A-c)

[0195] (R)-7-(5-(2-cyanopyridin-4-yl)oxazol-2-carboxamido)-5-azaspiro[2.4]heptane-5-carboxylic acid tert-butyl ester (Cpd-35A-b, 30 mg, 0.073 mmol) was dissolved in 1 mL of dichloromethane, and then 1 mL of trifluoroacetic acid was added. The reaction solution was stirred at 20 °C for 2 h. After the reaction was completed, the reaction solution was concentrated to obtain crude (R)-5-(2-cyanopyridin-4-yl)-N-(5-azaspiro[2.4]heptane-7-yl)oxazol-2-carboxamide trifluoroacetate (Cpd-35A-c, 15 mg), yield: 59%.

[0196] MS m / z (ESI): 384.1 (M+1)

[0197] Step 3: Preparation of (R)-N-(5-cyano-5-azaspiro[2.4]heptane-7-yl)-5-(2-cyanopyridin-4-yl)oxazol-2-carboxamide (Cpd-35A)

[0198] (R)-5-(2-cyanopyridin-4-yl)-N-(5-azaspiro[2.4]heptane-7-yl)oxazol-2-carboxamide trifluoroacetate (Cpd-35A-c, 15 mg, 0.048 mmol) was dissolved in dichloromethane, followed by the addition of N,N-diisopropylethylamine (18 mg, 0.154 mmol), and then cyanogen bromide (15 mg, 0.154 mmol). The reaction mixture was stirred at 20 °C for 1 h. After the reaction was completed, the reaction mixture was concentrated and purified by silica gel agar (petroleum ether / ethyl acetate = 1 / 1) to obtain the product (R)-N-(5-cyano-5-azaspiro[2.4]heptane-7-yl)-5-(2-cyanopyridin-4-yl)oxazol-2-carboxamide (Cpd-35A, 7.51 mg), yield: 46.50%.

[0199] MS m / z (ESI): 335.1 (M+1)

[0200] HPLC:100%(214nm),100%(254nm)

[0201] 1 H NMR (400MHz, DMSO) δ9.45(d,J=8.0Hz,1H),8.89(dd,J=5.2,0.7Hz,1H),8.50(d,J=0.8Hz,1H),8.34(d,J=8.8Hz,1H),8.09(dd,J=5.2,1.7Hz,1H ),4.30–4.19(m,1H),3.87(dd,J=10.0,6.6Hz,1H),3.73(d,J=9.2Hz,1H),3.54(dd,J=10.0,3.5Hz,1H),3.24(d,J=9.2Hz,1H),0.94–0.59(m,4H)

[0202] Example 7 7-(5-(2-cyanopyridin-4-yl)oxazol-2-carbonyl)-2,7-diazaspiro[4.4]nonane-2-nitrile (compound Cpd-36)

[0203] Step 1: Preparation of tert-butyl 7-(5-(2-cyanopyridin-4-yl)oxazol-2-carbonyl)-2,7-diazaspiro[4.4]nonane-2-carboxylate (Cpd-36a)

[0204] Ethyl 5-(2-cyanopyridin-4-yl)-1,3-oxazol-2-carboxylate (Cpd-35A-a, 30 mg, 0.123 mmol) and tert-butyl 2,7-diazaspiro[4.4]nonane-2-carboxylate (28 mg, 0.123 mmol) were dissolved in tetrahydrofuran (0.6 mL), and 1,8-diazabicyclo[5.4.0]undec-7-ene (150 mg, 0.986 mmol) was added. The reaction mixture was stirred for 16 h at room temperature under nitrogen protection. After the reaction was completed, the reaction mixture was poured into water and extracted with ethyl acetate (5 mL × 2). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated to dryness, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:2) to give tert-butyl 7-(5-(2-cyanopyridin-4-yl)oxazol-2-carbonyl)-2,7-diazaspiro[4.4]nonane-2-carboxylate (Cpd-36a, 40 mg), yield: 76%.

[0205] MS m / z (ESI): 368.0 (M-56+1)

[0206] Step 2: Preparation of 4-(2-(2,7-diazaspiro[4.4]nonane-2-carbonyl)oxazol-5-yl)-2-cyanopyridine trifluoroacetate (Cpd-36b)

[0207] 7-(5-(2-cyanopyridin-4-yl)oxazol-2-carbonyl)-2,7-diazaspiro[4.4]nonane-2-carboxylic acid tert-butyl ester (Cpd-36a, 40 mg, 0.0945 mmol) was dissolved in dichloromethane (1 mL), and trifluoroacetic acid (302 mg, 2.65 mmol) was added. The reaction mixture was stirred at 25 °C for 1 h. After the reaction was completed, the reaction mixture was concentrated to dryness to give 4-(2-(2,7-diazaspiro[4.4]nonane-2-carbonyl)oxazol-5-yl)-2-cyanopyridinium trifluoroacetate (Cpd-36b, 40 mg), yield: 96%.

[0208] MS m / z (ESI): 324.1 (M+1)

[0209] Step 3: Preparation of 7-(5-(2-cyanopyridin-4-yl)oxazol-2-carbonyl)-2,7-diazaspiro[4.4]nonane-2-nitrile (Cpd-36)

[0210] 4-(2-(2,7-diazaspiro[4.4]nonane-2-carbonyl)oxazol-5-yl)-2-cyanopyridine trifluoroacetate (Cpd-36b, 40 mg, 0.0915 mmol) was dissolved in dichloromethane (2 mL), and diisopropylethylamine (71 mg, 0.549 mmol) and cyanogen bromide (11 mg, 0.101 mmol) were added. The reaction mixture was stirred at 25 °C for 1 h. After the reaction was complete, water was added to the reaction mixture to adjust the pH to 5, and the mixture was extracted with dichloromethane, washed with brine, and concentrated to dryness. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:2) to give 7-(5-(2-cyanopyridine-4-yl)oxazol-2-carbonyl)-2,7-diazaspiro[4.4]nonane-2-onitrile (Cpd-36, 16.62 mg), yield: 52%.

[0211] MS m / z (ESI): 349.1 (M+1)

[0212] HPLC: 95.61% (214nm), 94.07% (254nm).

[0213] 1 H NMR (400MHz, DMSO) δ8.87(d,J=5.2Hz,1H),8.49(s,1H),8.34(s,1H),8.05(ddd,J=5.0,3.0,1.8Hz,1H),4.06(td,J=12.0,4.8Hz,1H),3.92(dd,J= 33.0,11.6Hz,1H),3.63(dd,J=11.1,7.2Hz,1H),3.55(d,J=23.0Hz,1H), 3.51–3.43(m,2H),3.40(d,J=9.4Hz,1H),3.36(s,1H),2.02–1.86(m,4H).

[0214] Example 8 (R)-N-(5-cyano-5-azaspiro[2.4]heptane-7-yl)-3-(4-fluorophenyl)imidazo[1,2-a]pyrazine-6-carboxamide (compound Cpd-30A)

[0215] Step 1: Preparation of imidazo[1,2-a]pyrazine-6-carboxylic acid ethyl ester (Cpd-30A-a)

[0216] To a solution of ethyl 5-aminopyrazine-2-carboxylate (1 g, 6 mmol) in ethanol (30 mL), 2-chloroacetaldehyde (7 mL, 36 mmol) was added, and the mixture was heated to 90 °C for 12 h. After the reaction was complete, the solution was concentrated under reduced pressure and purified by silica gel column chromatography (0%–10% methanol in dichloromethane) to give imidazo[1,2-a]pyrazine-6-carboxylate (Cpd-30A-a, 770 mg), with a yield of 63%.

[0217] MS m / z (ESI): 192.1 (M+1)

[0218] Step 2: Preparation of ethyl 3-bromoimidazolo[1,2-a]pyrazine-6-carboxylate (Cpd-30A-b)

[0219] Ethyl imidazo[1,2-a]pyrazine-6-carboxylate (Cpd-30A-a, 770 mg, 4 mmol) was dissolved in 10 mL of dichloromethane, and N-bromosuccinimide (1 g, 6 mmol) was added. The reaction mixture was stirred at 20 °C for 1 h. After the reaction was completed, the solution was concentrated under reduced pressure and purified by silica gel column chromatography (40%–50% ethyl acetate in petroleum ether) to give ethyl 3-bromoimidazo[1,2-a]pyrazine-6-carboxylate (Cpd-30A-b, 900 mg), yield: 74%.

[0220] MS m / z (ESI): 300.0 (M+1)

[0221] Step 3: Preparation of ethyl 3-(4-fluorophenyl)imidazo[1,2-a]pyrazine-6-carboxylate (Cpd-30A-c)

[0222] To a mixed solution of ethyl 3-bromoimidoz[1,2-a]pyrazine-6-carboxylic acid (Cpd-30A-b, 900 mg, 3.3 mmol) and (4-fluorophenyl)boronic acid (699 mg, 5 mmol) in 1,4-dioxane (10 mL) and water (1 mL), palladium dichloride (244 mg, 0.33 mmol) and potassium carbonate (921 mg, 6.66 mmol) were added, and the mixture was heated to 90 °C for 12 h. After the reaction was completed, the solution was concentrated under reduced pressure and purified by silica gel column chromatography (80%–90% ethyl acetate in petroleum ether) to give 3-(4-fluorophenyl)imidoz[1,2-a]pyrazine-6-carboxylic acid (Cpd-30A-c, 80 mg), in 10% yield.

[0223] MS m / z (ESI): 286.1 (M+1)

[0224] Step 4: Preparation of (R)-7-(3-(4-fluorophenyl)imidazo[1,2-a]pyrazin-6-carboxamido)-5-azaspiro[2,4]heptane-5-carboxylic acid tert-butyl ester (Cpd-30A-d)

[0225] 3-(4-fluorophenyl)imidazo[1,2-a]pyrazine-6-carboxylic acid (Cpd-30A-c, 80 mg, 0.3 mmol) was dissolved in dichloromethane, followed by the addition of (R)-7-amino-5-azaspiro[2,4]heptane-5-carboxylic acid tert-butyl ester (66 mg, 0.3 mmol), HATU (177 mg, 0.5 mmol), and N,N-diisopropylethylamine (60 mg, 0.5 mmol). The reaction mixture was stirred at 20 °C for 1 h. After the reaction was completed, the solution was concentrated under reduced pressure and subjected to silica gel column chromatography (50%–60% ethyl acetate in petroleum ether) to obtain (R)-7-(3-(4-fluorophenyl)imidazo[1,2-a]pyrazine-6-carboxamido)-5-azaspiro[2,4]heptane-5-carboxylic acid tert-butyl ester (Cpd-30A-d, 50 mg), yield: 32%.

[0226] MS m / z (ESI): 452.2 (M+1)

[0227] Step 5: Preparation of (R)-3-(4-fluorophenyl)-N-(5-azaspiro[2.4]heptane-7-yl)imidazo[1,2-a]pyrazine-6-carboxamide trifluoroacetate (Cpd-30A-e)

[0228] (R)-7-(3-(4-fluorophenyl)imidazo[1,2-a]pyrazin-6-carboxamido)-5-azaspiro[2,4]heptane-5-carboxylic acid tert-butyl ester (Cpd-30A-d, 50 mg, 0.11 mmol) was dissolved in 1 mL of dichloromethane, followed by the addition of 1 mL of trifluoroacetic acid. The reaction mixture was stirred at 20 °C for 2 h. After the reaction was completed, the reaction mixture was concentrated to obtain crude (R)-3-(4-fluorophenyl)-N-(5-azaspiro[2,4]heptane-7-yl)imidazo[1,2-a]pyrazin-6-carboxamide trifluoroacetate (Cpd-30A-e, 40 mg), yield: 92%.

[0229] MS m / z (ESI): 352.2 (M+1)

[0230] Step 6: Preparation of (R)-N-(5-cyano-5-azaspiro[2.4]heptane-7-yl)-3-(4-fluorophenyl)imidazo[1,2-a]pyrazine-6-carboxamide (Cpd-30A)

[0231] (R)-3-(4-fluorophenyl)-N-(5-azaspiro[2.4]heptane-7-yl)imidazo[1,2-a]pyrazin-6-carboxamide (Cpd-30A-e, 40 mg, 0.11 mmol) was dissolved in dichloromethane, followed by the addition of N,N-diisopropylethylamine (44 mg, 0.34 mmol), and then cyanogen bromide (36 mg, 0.34 mmol). The reaction mixture was stirred at 20 °C for 1 h. After the reaction was completed, the solution was concentrated under reduced pressure and then subjected to silica gel column chromatography (60%–70% ethyl acetate in petroleum ether) to obtain (R)-N-(5-cyano-5-azaspiro[2.4]heptane-7-yl)-3-(4-fluorophenyl)imidazo[1,2-a]pyrazin-6-carboxamide (Cpd-30A, 20.96 mg), yield: 48%.

[0232] MS m / z (ESI): 377.1 (M+1)

[0233] HPLC:100%(214nm),99.75%(254nm)

[0234] 1 H NMR (400MHz, DMSO) δ9.21(d,J=1.2Hz,1H),8.97(d,J=8.0Hz,1H),8.86(d,J=1.2Hz ,1H),8.15(s,1H),7.85–7.80(m,2H),7.49(t,J=8.8Hz,2H),4.31–4.25(m,1H),3.8 4(dd,J=9.8,6.6Hz,1H),3.73(d,J=9.2Hz,1H),3.61(dd,J=9.8,4.0Hz,1H),3.27( d,J=9.2Hz,1H),0.82–0.77(m,1H),0.71(dd,J=8.6,4.7Hz,2H),0.64–0.59(m,1H).

[0235] Example 9 N-(3-cyano-3-azabicyclo[3.1.0]hexane-1-yl)-5-(2-cyanopyridin-4-yl)oxazol-2-carboxamide (compound Cpd-41)

[0236] Step 1: Preparation of 1-(5-(2-cyanopyridin-4-yl)oxazol-2-carboxamido)-3-azabicyclo[3.1.0]hexane-3-carboxylic acid tert-butyl ester (Cpd-41a)

[0237] Ethyl 5-(2-cyanopyridin-4-yl)-1,3-oxazol-2-carboxylate (Cpd-35A-a, 200 mg, 0.819 mmol) was dissolved in 3 mL of tetrahydrofuran, followed by the addition of 1,8-diazabicycloundec-7-ene (373 mg, 2.39 mmol) and 1-amino-3-azabicyclo[3.1.0]hexane-3-onitrile (186 mg, 0.899 mmol). The reaction mixture was stirred at 20 °C for 16 h. After the reaction was completed, the reaction mixture was concentrated and purified by silica gel agar plate preparation (petroleum ether / ethyl acetate = 1 / 1) to obtain the product 1-(5-(2-cyanopyridin-4-yl)oxazol-2-carboxamido)-3-azabicyclo[3.1.0]hexane-3-carboxylate tert-butyl ester (Cpd-41a, 130 mg), yield: 40%.

[0238] MS m / z (ESI): 340.0 (M-55)

[0239] Step 2: Preparation of N-(3-azabicyclo[3.1.0]hexane-1-yl)-5-(2-cyanopyridin-4-yl)oxazol-2-carboxamide trifluoroacetate (Cpd-41b)

[0240] 1-(5-(2-cyanopyridin-4-yl)oxazol-2-carboxamido)-3-azabicyclo[3.1.0]hexane-3-carboxylic acid tert-butyl ester (Cpd-41a, 130 mg, 0.313 mmol) was dissolved in 2 mL of dichloromethane, followed by the addition of 2 mL of trifluoroacetic acid. The reaction mixture was stirred at 20 °C for 2 h. After the reaction was completed, the reaction mixture was concentrated to obtain crude N-(3-azabicyclo[3.1.0]hexane-1-yl)-5-(2-cyanopyridin-4-yl)oxazol-2-carboxamide trifluoroacetate (Cpd-41b, 80 mg), yield: 82.4%.

[0241] MS m / z (ESI): 296.1 (M+1)

[0242] Step 3: Preparation of N-(3-cyano-3-azabicyclo[3.1.0]hexane-1-yl)-5-(2-cyanopyridin-4-yl)oxazol-2-carboxamide (Cpd-41)

[0243] N-(3-azabicyclo[3.1.0]hexan-1-yl)-5-(2-cyanopyridin-4-yl)oxazol-2-carboxamide trifluoroacetate (Cpd-41b, 80 mg, 0.254 mmol) was dissolved in dichloromethane, followed by the addition of N,N-diisopropylethylamine (97.2 mg, 0.831 mmol), and then cyanogen bromide (81 mg, 0.831 mmol). The reaction mixture was stirred at 20 °C for 1 h. After the reaction was completed, the reaction mixture was concentrated and purified by silica gel agar (petroleum ether / ethyl acetate = 1 / 1) to obtain N-(3-cyano-3-azabicyclo[3.1.0]hexan-1-yl)-5-(2-cyanopyridin-4-yl)oxazol-2-carboxamide (Cpd-41, 70 mg), yield: 81%.

[0244] Cpd-41 (70 mg) was chirally separated (mobile phase: CO2 / MeOH [0.2% NH3 (7M Solution in MeOH)] = 50 / 50) (separation conditions: instrument: Waters Acquity UPCC; chiral column: Daicel CHIRALCEL ID, 250 mm × 30 mm ID, 10 μm; mobile phase: A / B: CO2 / MeOH (0.2% NH3 (7M MeOH solution)) = 60 / 40) to obtain Cpd-41A (first peak, retention time: 1.947 min, 19.23 mg) and Cpd-41B (last peak, retention time: 2.931 min, 16.71 mg).

[0245] Cpd-41A:

[0246] MS m / z (ESI): 321.0 (M+1)

[0247] HPLC:100%(214nm),100%(254nm)

[0248] 1 H NMR (400MHz, DMSO) δ9.68(s,1H),8.89(d,J=5.2Hz,1H),8.51(s,1H),8.32(s,1H),8.10(dd,J=5.2,1.6Hz,1H),3.72(dd,J=9.6,3.9Hz,1 H),3.61(d,J=9.2Hz,1H),3.55(d,J=9.2Hz,1H),3.39(d,J=9.6Hz,1H),1.94–1.79(m,1H),1.24(t,J=7.2Hz,2H),0.95(t,J=5.6Hz,1H).

[0249] Cpd-41B:

[0250] MS m / z (ESI): 321.1 (M+1)

[0251] HPLC:100%(214nm),100%(254nm)

[0252] 1 H NMR (400MHz, DMSO) δ9.68(s,1H),8.89(d,J=5.2Hz,1H),8.51(s,1H),8.32(s,1H),8.10(dd,J=5.2,1.6Hz,1H),3.72(dd,J=9.6,3.8Hz,1 H),3.61(d,J=9.2Hz,1H),3.55(d,J=9.6Hz,1H),3.39(d,J=9.6Hz,1H),1.92–1.82(m,1H),1.24(t,J=7.2Hz,2H),0.95(t,J=5.6Hz,1H).

[0253] Example 10 N-(3-cyano-4-methyl-3-azabicyclo[3.1.0]hexane-1-yl)-5-(2-cyanopyridin-4-yl)oxazol-2-carboxamide (compound Cpd-42)

[0254] Step 1: Preparation of tert-butyl butyronitrile-3-en-2-yl(cyanomethyl)carbamate (Cpd-42a)

[0255] 2 g (0.0128 mol) of tert-butyl (cyanomethyl) carbamate was dissolved in 10 mL of N,N-dimethylformamide, and sodium hydride (0.82 g, 0.02 mol, 60% aqueous solution) was added. The reaction mixture was stirred at 0 °C for 1 h. Then 0.88 g (0.01 mol) of 3-chlorobut-1-ene was added, and the reaction was carried out at room temperature for 4 h. After the reaction was completed, ice water was poured into the reaction mixture, and the mixture was extracted with ethyl acetate (5 mL × 2). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated to dryness, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 4:1) to give tert-butyl butyronitrile (Cpd-42a, 1.1 g), yield: 41%.

[0256] MS m / z (ESI): 211.1 (M+1)

[0257] Step 2: Preparation of 1-amino-4-methyl-3-azabicyclo[3.1.0]hexane-3-carboxylic acid tert-butyl ester (Cpd-42b)

[0258] Butyl-3-en-2-yl(cyanomethyl)carbamate tert-butyl ester (Cpd-42a, 500 mg, 2.3794 mmol) was dissolved in tetrahydrofuran (10 mL), and triisopropoxy(methyl)titanium (6 mL, 1 mol / L) was slowly added. The mixture was stirred at room temperature for 10 min, and then cyclohexylmagnesium bromide (2.4 mL, 2 mol / L) was slowly added. The reaction was allowed to proceed for 16 h at room temperature. After the reaction was complete, the solvent was removed by rotary evaporation, and the product was purified by silica gel column chromatography (dichloromethane / methanol = 5:1) to give 1-amino-4-methyl-3-azabicyclo[3.1.0]hexane-3-carboxylic acid tert-butyl ester (Cpd-42b, 200 mg), yield: 40%.

[0259] MS m / z (ESI): 157.2 (M+1-56)

[0260] Step 3: Preparation of 5-(2-cyanopyridin-4-yl)oxazol-2-carboxylic acid (Cpd-42c)

[0261] 1-Amino-4-methyl-3-azabicyclo[3.1.0]hexane-3-carboxylic acid tert-butyl ester (Cpd-35A-a, 200 mg, 0.8223 mmol) was dissolved in a tetrahydrofuran / water mixture of 5 / 2 (7 mL), and triethylamine (250 mg, 2.4669 mmol) was added. The reaction was carried out at 50 °C for 5 h. After the reaction was completed, the solvent was removed by vortexing, and the product was directly dried to obtain 5-(2-cyanopyridin-4-yl)oxazol-2-carboxylic acid (Cpd-42c). The crude product was used directly in the next step.

[0262] MS m / z (ESI): 216.1 (M+1)

[0263] Step 4: Preparation of 1-(5-(2-cyanopyridin-4-yl)oxazol-2-carboxamido)-4-methyl-3-azabicyclo[3.1.0]hexane-3-carboxylic acid tert-butyl ester (Cpd-42d)

[0264] 5-(2-cyanopyridin-4-yl)azole-2-carboxylic acid (Cpd-42c, 200 mg, 0.9295 mmol, crude) was dissolved in N,N-dimethylformamide (5 mL), and HATU (530 mg, 1.3943 mmol), N,N-diisopropylethylamine (360 mg, 2.7885 mmol), and tert-butyl 1-amino-4-methyl-3-azabicyclo[3.1.0]hexane-3-carboxylic acid (Cpd-42b, 395 mg, 1.859 mmol) were added. The reaction was carried out at room temperature for 1 h. After the reaction was complete, the mixture was diluted with water, extracted with ethyl acetate, and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 2:1) to give 1-(5-(2-cyanopyridin-4-yl)oxazol-2-carboxamido)-4-methyl-3-azabicyclo[3.1.0]hexane-3-carboxylic acid tert-butyl ester (Cpd-42d, 120 mg), yield: 28%.

[0265] MS m / z (ESI): 310.8 (M+1-100)

[0266] Step 5: Preparation of 5-(2-cyanopyridin-4-yl)-N-(4-methyl-3-azabicyclo[3.1.0]hexane-1-yl)oxazol-2-carboxamide trifluoroacetate (Cpd-42e)

[0267] 1-(5-(2-cyanopyridin-4-yl)oxazol-2-carboxamido)-4-methyl-3-azabicyclo[3.1.0]hexane-3-carboxylic acid tert-butyl ester (Cpd-42d, 110 mg, 0.268 mmol) was dissolved in dichloromethane (3 mL), and trifluoroacetic acid (1 mL) was added. The reaction was carried out at room temperature for 1 h. After the reaction was completed, the solvent was directly removed by vortexing to obtain 5-(2-cyanopyridin-4-yl)-N-(4-methyl-3-azabicyclo[3.1.0]hexane-1-yl)oxazol-2-carboxamide trifluoroacetate (Cpd-42e), and the crude product was used directly in the next step.

[0268] MS m / z (ESI): 310.8 (M+1)

[0269] Step 6: Preparation of N-(3-cyano-4-methyl-3-azabicyclo[3.1.0]hexane-1-yl)-5-(2-cyanopyridin-4-yl)oxazol-2-carboxamide (Cpd-42)

[0270] 5-(2-cyanopyridin-4-yl)-N-(4-methyl-3-azabicyclo[3.1.0]hexan-1-yl)oxazol-2-carboxamide trifluoroacetate (Cpd-42e, 110 mg, 0.291 mmol, crude) was dissolved in tetrahydrofuran (3 mL), and potassium carbonate (101 mg, 0.7275 mmol) and cyanogen bromide (62 mg, 0.582 mmol) were added. The reaction was carried out at room temperature for 1 h. After the reaction was completed, the mixture was extracted with water and ethyl acetate, the organic phase was collected, evaporated to dryness, and the residue was subjected to silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 2) to give N-(3-cyano-4-methyl-3-azabicyclo[3.1.0]hexan-1-yl)-5-(2-cyanopyridin-4-yl)oxazol-2-carboxamide (Cpd-42, 30 mg).

[0271] MS m / z (ESI): 335.1 (M+1)

[0272] HPLC: 94.24% (214nm), 98.91% (254nm).

[0273] Compound Cpd-42 was resolved by SFC (resolution method: instrument: Waters Acquity UPCC; chiral column: Daicel CHIRALPAK ID_3, 3.0*150mm, 3μm; mobile phase: A / B:CO2 / MeOH (0.1% DEA)=60 / 40; flow rate: 1.5ml / min; temperature: 37℃) to obtain Cpd-42A (retention time: 1.560min, 9.92mg); Cpd-42B (retention time: 1.947min, 8.38mg); Cpd-42C (retention time: 2.766min, 7.41mg).

[0274] Cpd-42A was separated by SFC (Separation method: Instrument: Waters Acquity UPCC, Chiral column: Daicel CHIRALPAK OX_3, 3*150mm, 3μm, Mobile phase: A / B:CO2 / MeOH (0.1% DEA)=60 / 40, Flow rate: 1.5ml / min, Temperature: 37℃) to obtain Cpd-42A1 (retention time: 1.598min, 1.16mg) and Cpd-42A2 (retention time: 1.982min, 1.11mg).

[0275] Cpd-42A1:

[0276] MS m / z (ESI): 335.1 (M+1)

[0277] HPLC: 78.11% (214nm), 96.72% (254nm).

[0278] 1 H NMR(400MHz,dmso)δ9.65(s,1H),8.87(d,J=5.1Hz,1H),8.49(s,1H),8.30(s,1H),8.08(dd,J=5.2,1.6Hz,1H),3.65(dd,J=15.0,7.8Hz,2H),3.51(d,J=8.9Hz,1H),1.97(dd,J=14.5,6.9Hz,1H),1.65(dd,J=8.6,5.1Hz,1H),1.40(d,J=6.4Hz,3H),0.91(t,J=5.5Hz,1H).

[0279] Cpd-42A2:

[0280] MS m / z(ESI):335.1(M+1)

[0281] HPLC:87.10%(214nm),96.90%(254nm).

[0282] 1 H NMR(400MHz,dmso)δ9.66(s,1H),8.87(d,J=5.1Hz,1H),8.49(s,1H),8.30(s,1H),8.08(d,J=3.9Hz,1H),3.65(dd,J=15.1,7.8Hz,2H),3.51(d,J=8.9Hz,1H),2.04–1.91(m,1H),1.65(dd,J=8.5,5.1Hz,1H),1.40(d,J=6.4Hz,3H),0.91(t,J=5.5Hz,1H).

[0283] Cpd-42B:

[0284] MS m / z(ESI):335.1(M+1)

[0285] HPLC:81.60%(214nm),92.45%(254nm).

[0286] 1 H NMR(400MHz,MeOD)δ8.80(d,J=5.1Hz,1H),8.33(s,1H),8.06(s,1H),8.04(d,J=5.1Hz,1H),4.32(d,J=3.4Hz,1H),4.16(dd,J=11.5,6.2Hz,2H),2.08–1.98(m,3H),1.16–1.07(m,2H).

[0287] Cpd-42C:

[0288] MS m / z (ESI): 335.1 (M+1)

[0289] HPLC: 76.82% (214nm), 93.03% (254nm).

[0290] 1 H NMR(400MHz,MeOD)δ8.80(d,J=5.2Hz,1H),8.33(s,1H),8.06(s,1H),8.04(d,J=5.1Hz,1H),4. 21–4.08(m,1H),3.71(s,2H),2.04(s,1H),1.31(t,J=7.2Hz,3H),1.11(dd,J=17.5,6.9Hz,2H).

[0291] Example 11 N-((2-cyano-2-azabicyclo[3.1.0]hexane-5-yl)methyl)-5-(2-cyanopyridin-4-yl)oxazol-2-carboxamide (compound Cpd-43)

[0292] Step 1: Preparation of 5-((1,3-dioxoisoindol-2-yl)methyl)-2-azabicyclo[3.1.0]hexane-2-carboxylic acid tert-butyl ester (Cpd-43a)

[0293] Under a nitrogen atmosphere, phthalimide (828 mg, 5.62 mmol) and triphenylphosphine (1.48 g, 5.63 mmol) were added to an ultradry tetrahydrofuran (30 mL) solution of 5-(hydroxymethyl)-2-azabicyclo[3.1.0]hexane-2-carboxylic acid tert-butyl ester (1 g, 4.69 mmol) (30 mL). Then, diisopropyl azodicarbonate (1.90 g, 9.38 mmol) was slowly added dropwise in an ice bath, and the reaction mixture was stirred at room temperature for 16 h. After the reaction was complete, the mixture was quenched with water and extracted with ethyl acetate (3 × 50 mL). The organic phase was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether / ethyl acetate = 10 / 1) to obtain 5-((1,3-dioxoisoindol-2-yl)methyl)-2-azabicyclo[3.1.0]hexane-2-carboxylic acid tert-butyl ester (Cpd-43a, 500 mg), yield: 31%.

[0294] MS m / z (ESI): 365.0 (M+Na).

[0295] Step 2: Preparation of 5-(aminomethyl)-2-azabicyclo[3.1.0]hexane-2-carboxylic acid tert-butyl ester (Cpd-43b)

[0296] Hydrazine hydrate (67 mg, 1.31 mmol) was added to a 3 mL ethanol solution of 5-((1,3-dioxoisoindol-2-yl)methyl)-2-azabicyclo[3.1.0]hexane-2-carboxylic acid tert-butyl ester (Cpd-43a, 150 mg, 0.44 mmol). The reaction mixture was stirred at 40 °C for 3 h. After the reaction was complete, the mixture was filtered directly, and the filtrate was concentrated and purified by column chromatography (mobile phase: water / acetonitrile = 5 / 1) to give 5-(aminomethyl)-2-azabicyclo[3.1.0]hexane-2-carboxylic acid tert-butyl ester (Cpd-43b, 50 mg), yield: 54%.

[0297] MS m / z(ESI): 140.1(M+1-56-17).

[0298] Step 3: Preparation of 5-((5-(2-cyanopyridin-4-yl)oxazol-2-carboxamido)methyl)-2-azabicyclo[3.1.0]hexane-2-carboxylic acid tert-butyl ester (Cpd-43c)

[0299] To a solution of 5-(2-cyanopyridin-4-yl)oxazol-2-carboxylic acid (Cpd-42c, 20 mg, 0.08 mmol) in N,N-dimethylformamide (5 mL), tert-butyl 5-(aminomethyl)-2-azabicyclo[3.1.0]hexane-2-carboxylic acid (Cpd-43b, 50 mg, 0.23 mmol), N,N,N',N'-tetramethyl-o-(7-azabenzotriazol-1-yl)hexafluorophosphate (63 mg, 0.16 mmol), and N,N-diisopropylethylamine (32 mg, 0.25 mmol) were added. The reaction mixture was stirred at 25 °C for 1 h. After the reaction was completed, the reaction was quenched with water, extracted with ethyl acetate (10 mL × 3), dried over anhydrous sodium sulfate on the organic phase, and the filtrate was concentrated and purified by silica gel column chromatography (mobile phase: petroleum ether / ethyl acetate = 1 / 1). The purified product was 5-((5-(2-cyanopyridin-4-yl)oxazol-2-carboxamido)methyl)-2-azabicyclo[3.1.0]hexane-2-carboxylic acid tert-butyl ester (Cpd-43c, 18 mg), yield: 54%.

[0300] MS m / z(ESI): 310.2(M+1-100).

[0301] Step 4: Preparation of N-((2-azabicyclo[3.1.0]hexane-5-yl)methyl)-5-(2-cyanopyridin-4-yl)oxazol-2-carboxamide trifluoroacetate (Cpd-43d)

[0302] 5-((5-(2-cyanopyridin-4-yl)oxazol-2-carboxamido)methyl)-2-azabicyclo[3.1.0]hexane-2-carboxylic acid tert-butyl ester (Cpd-43c, 18 mg, 0.04 mmol) was dissolved in a dichloromethane:trifluoroacetic acid = 3:1 (2 mL) solution. The reaction mixture was stirred at room temperature for 1 h. After the reaction was completed, the mixture was concentrated to give N-((2-azabicyclo[3.1.0]hexane-5-yl)methyl)-5-(2-cyanopyridin-4-yl)oxazol-2-carboxamide trifluoroacetate (Cpd-43d, 20 mg).

[0303] MS m / z (ESI): 310.1 (M+1).

[0304] Step 5: Preparation of N-((2-cyano-2-azabicyclo[3.1.0]hexane-5-yl)methyl)-5-(2-cyanopyridin-4-yl)oxazol-2-carboxamide (Cpd-43)

[0305] Under ice bath conditions, potassium carbonate (23 mg, 0.17 mmol) was added to a stirred solution of N-((2-azabicyclo[3.1.0]hexan-5-yl)methyl)-5-(2-cyanopyridin-4-yl)oxazol-2-carboxamide trifluoroacetate (Cpd-43d, 20 mg, 0.04 mmol) in tetrahydrofuran (3 mL). After 10 minutes, cyanogen bromide (7 mg, 0.06 mmol) was added dropwise. After the addition was complete, the reaction was brought to room temperature and stirred for 1 hour. After the reaction was complete, water (10 mL) was added to quench the reaction, and the mixture was extracted with dichloromethane (10 mL × 2), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by high performance liquid chromatography (Waters MS-triggered Prep-LC with SQD2 detector, column: Xbridge 5μm C18 150×19mm; mobile phase 1: water (containing 0.1% FA); mobile phase 2: acetonitrile; 9-minute gradient, gradient ratio: acetonitrile phase 45%-55%, flow rate: 20mL / min) to obtain N-((2-cyano-2-azabicyclo[3.1.0]hexane-5-yl)methyl)-5-(2-cyanopyridin-4-yl)oxazol-2-carboxamide (Cpd-43, 2.5mg), yield: 18%.

[0306] MS m / z (ESI): 335.1 (M+1).

[0307] HPLC: 96.82% (214nm), 96.69% (254nm).

[0308] 1H NMR (400MHz, CDCl3) δ8.82(d,J=5.2Hz,1H),8.01(s,1H),7.84(dd,J=5.2,1.6Hz,1H),7.72(s,1H),3.64–3 .61(m,3H),3.31–3.25(m,1H),3.14–3.07(m,1H),2.15–2.11(m,2H),1.18–1.15(m,1H),0.95–0.91(m,1H).

[0309] Example 12 7-(5-(2-cyanopyridin-4-yl)oxazol-2-carbonyl)-3-methyl-2,7-diazaspiro[4.4]nonane-2-nitrile (compound Cpd-44)

[0310] Step 1: Preparation of tert-butyl 4-(2-ethoxy-2-oxoethylidene)-2-methylpyrrolidine-1-carboxylate (Cpd-44a)

[0311] 5 g of tert-butyl 2-methyl-4-oxopyrrolidine-1-carboxylate (0.0251 mol) was dissolved in toluene (50 mL), and ethyl 2-(triphenyl-λ) was added. 5 13.12 g (0.03765 ​​mol) of phosphono(2-phosphine) acetate was refluxed and stirred for 12 h. After the reaction was complete, the solution was evaporated to dryness. The product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1) to give tert-butyl 4-(2-ethoxy-2-oxoethylene)-2-methylpyrrolidine-1-carboxylate (Cpd-44a, 6 g), in 88.84% yield.

[0312] MS m / z(ESI): 214.1(M+1-56).

[0313] Step 2: Preparation of tert-butyl 4-(2-ethoxy-2-oxoethyl)-2-methyl-4-(nitromethyl)pyrrolidine-1-carboxylate (Cpd-44b)

[0314] tert-butyl 4-(2-ethoxy-2-oxoethylene)-2-methylpyrrolidine-1-carboxylate (Cpd-44a, 2 g, 0.0074 mol) was dissolved in dimethyl sulfoxide (20 mL), and nitromethane (1.36 g, 0.0222 mol) and cesium carbonate (4.82 g, 0.0148 mol) were added. The mixture was stirred at 80 °C for 3 h. After the reaction was complete, the mixture was extracted with ethyl acetate / water, and the organic phases were combined and evaporated to dryness. The solution was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 5 / 1) to obtain tert-butyl 4-(2-ethoxy-2-oxoethyl)-2-methyl-4-(nitromethyl)pyrrolidine-1-carboxylate (Cpd-44b, 1.7 g), in 68.92% yield.

[0315] MS m / z (ESI): 275 (M+1-56).

[0316] Step 3: Preparation of tert-butyl-3-methyl-8-oxo-2,7-diazaspiro[4.4]nonane-2-carboxylate (Cpd-44c)

[0317] tert-butyl 4-(2-ethoxy-2-oxoethyl)-2-methyl-4-(nitromethyl)pyrrolidine-1-carboxylate (Cpd-44b, 1.7 g, 0.0051 mol) was dissolved in methanol (20 mL), and 10% palladium on carbon (0.17 g, 0.00153 mol) and 4M ammonia / methanol (1.5 mL) were added. The mixture was stirred at 40 °C for 5 h. After the reaction was completed, the mixture was filtered and evaporated to dryness. The tert-butyl 3-methyl-8-oxo-2,7-diazaspiro[4.4]nonane-2-carboxylate (Cpd-44c, 0.75 g) was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 2) to obtain tert-butyl 3-methyl-8-oxo-2,7-diazaspiro[4.4]nonane-2-carboxylate (Cpd-44c, 0.75 g), with a yield of 56.86%.

[0318] MS m / z(ESI): 199.2(M+1-56).

[0319] Step 4: Preparation of tert-butyl-3-methyl-2,7-diazaspiro[4.4]nonane-2-carboxylate (Cpd-44d)

[0320] tert-butyl-3-methyl-8-oxo-2,7-diazaspiro[4.4]nonane-2-carboxylate (Cpd-44c, 400 mg, 1.5728 mmol) was dissolved in 1 M tetrahydrofuran borane (10 mL) and refluxed with stirring for 8 h. After the reaction was completed, methanol was added to quench the reaction, followed by water, and refluxed for 5 h. Tert-butyl-3-methyl-2,7-diazaspiro[4.4]nonane-2-carboxylate (Cpd-44d, 160 mg) was obtained by separation and purification by reverse-phase column chromatography (water / acetonitrile = 1 / 5), with a yield of 42.33%.

[0321] MS m / z (ESI): 241.2 (M+1).

[0322] Step 5: Preparation of tert-butyl 7-(5-(2-cyanopyridin-4-yl)oxazol-2-carbonyl)-3-methyl-2,7-diazaspiro[4.4]nonane-2-carboxylate (Cpd-44e)

[0323] tert-butyl-3-methyl-2,7-diazaspiro[4.4]nonane-2-carboxylate (Cpd-44d, 160 mg, 0.6657 mmol) was dissolved in N,N-dimethylformamide (5 mL), and 5-(2-cyanopyridin-4-yl)oxazol-2-carboxylic acid (Cpd-42c, 143.24 mg, 0.6657 mmol), tetramethylchlorourea hexafluorophosphate (373.56 mg, 1.3314 mmol), and N-methylimidazolium (109.32 mg, 1.3314 mmol) were added. The mixture was stirred at 25 °C for 1 h. After the reaction was complete, ethyl acetate / water was added for extraction. The product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 2) to obtain tert-butyl 7-(5-(2-cyanopyridin-4-yl)oxazol-2-carbonyl)-3-methyl-2,7-diazaspiro[4.4]nonane-2-carboxylate (Cpd-44e, 160 mg), with a yield of 54.93%.

[0324] MS m / z(ESI): 382.1(M+1-56).

[0325] Step 6: Preparation of 4-(2-(8-methyl-2,7-diazaspiro[4.4]nonane-2-carbonyl)oxazol-5-yl)pyridinium trifluoroacetate (Cpd-44f)

[0326] Tert-butyl 7-(5-(2-cyanopyridin-4-yl)oxazol-2-carbonyl)-3-methyl-2,7-diazaspiro[4.4]nonane-2-carboxylate (Cpd-44e, 160 mg, 0.3657 mmol) was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (416.97 mg, 3.657 mmol) was added. The mixture was stirred at 25 °C for 3 h. After the reaction was completed, the solution was evaporated to dryness to obtain the crude product 4-(2-(8-methyl-2,7-diazaspiro[4.4]nonane-2-carbonyl)oxazol-5-yl)pyridinium trifluoroacetate (Cpd-44f, 60 mg), with a yield of 48.62%.

[0327] MS m / z (ESI): 338.1 (M+1).

[0328] Step 7: Preparation of 7-(5-(2-cyanopyridin-4-yl)oxazol-2-carbonyl)-3-methyl-2,7-diazaspiro[4.4]nonane-2-nitrile (Cpd-44)

[0329] 4-(2-(8-methyl-2,7-diazaspiro[4.4]nonane-2-carbonyl)oxazol-5-yl)pyridinium trifluoroacetate (Cpd-44f, 60 mg, 0.1778 mmol) was dissolved in tetrahydrofuran (5 mL), and N-ethyldiisopropylamine (45.96 mg, 0.3556 mmol) and bromoacetonitrile (22.60 mg, 0.21336 mmol) were added. The mixture was stirred at 25 °C for 1 h. After the reaction was complete, ethyl acetate / water was added for extraction. The compound 7-(5-(2-cyanopyridin-4-yl)oxazol-2-carbonyl)-3-methyl-2,7-diazaspiro[4.4]nonane-2-nitrile (Cpd-44, 43 mg) was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 1 / 5) with a yield of 66.76%.

[0330] MS m / z (ESI): 363.1 (M+1)

[0331] HPLC: 98.16% (214nm), 98.82% (254nm).

[0332] 1 H NMR (400MHz, DMSO) δ8.87(d,J=5.2Hz,1H),8.49(d,J=3.3Hz,1H),8.34(d,J=2.0Hz,1H),8.05(dt,J=5.1,1.9Hz,1H),4.06-3.73(m,3H),3.59(d d,J=9.5,6.1Hz,1H),3.49-3.42(m,2H),2.12(ddd,J=12.5,6.4,3.3Hz,1H),2.03-1.90(m,2H),1.57-1.48(m,1H),1.25(dd,J=8.5,6.2Hz,4H).

[0333] The compounds listed in Table 1 below were prepared using conditions similar to those in the examples described above. The structural characterization data of these compounds are also listed in Table 1.

[0334] Table 1

[0335] Biological evaluation

[0336] Test Example 1: Inhibition of deubiquitinating enzyme USP30

[0337] The inhibitory activity of the compound against USP30 was tested using a deubiquitinase fluorescent reporter assay. The highest concentration tested was 1000 nM, with 3-fold serial dilutions, 10 concentrations, and replicate detection.

[0338] 1.1 Experimental Materials

[0339] 1.1.1 Reagents and Consumables

[0340] 1.1.2 Instruments

[0341] 1.2 Experimental Procedure

[0342] a) Dilute USP30 and Ub-Rh110 to working concentrations as shown in the table below:

[0343] b) Using an Echo pipetting workstation, serially diluted compounds were transferred into a 384-well test plate, with a maximum concentration of 1000 nM, for a total of 10 concentrations. The plates were centrifuged at 1000 rpm for 30 seconds. For this test method, an Echo was used to transfer an equal volume of DMSO to the test compound, with High and Low controls set for data analysis.

[0344] High control(No compound):5μL USP30+5μL Ub-Rh110

[0345] Low control(No compound):5μL Ub-Rh110+5μL DMSO

[0346] c) Add 5 μL of working concentration of USP30 to each well, centrifuge, and incubate at 25°C for 25 min.

[0347] d) Add 5 μL of working concentration of Ub-Rh110 to each well, centrifuge, and incubate at 25°C for 2 h.

[0348] e) Use a plate reader to read the fluorescence signal of each well (Ex480 / Em540), and then calculate the inhibition percentage of each compound well according to the formula "Inhibition rate per well = (Average signal value of high control - Signal value per well) / (Average signal value of high control - Average signal value of low control) * 100%". Use a four-parameter logistic regression model to fit the activation rate-concentration curve, and finally calculate the IC50 of the compound. 50 Numerical value.

[0349] Experimental results: The IC50 values ​​of the representative compounds in this paper for their inhibitory activity against USP30 are... 50 The values ​​are shown in Table 2.

[0350] Table 2 shows the IC50 values ​​of the representative compounds in this paper for their inhibitory activity against USP30. 50 numerical values ++++:IC50 ≤1nM; +++:1nM <IC 50 ≤10nM; ++:10nM <IC 50 ≤100nM; +:100nM <IC 50 ≤1000nM

[0351] Experimental conclusion: The above representative compounds can effectively inhibit the activity of deubiquitinase USP30.

[0352] The embodiments of the technical solution of the present invention have been described above by way of example. It should be understood that the protection scope of the present invention is not limited to the above embodiments. Any modifications, equivalent substitutions, improvements, etc., made by those skilled in the art within the spirit and principles of the present invention should be included within the protection scope of the claims of this application.

Claims

1. The compound represented by formula (I), its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt, or prodrug compound: in, X1, X2, X3, X4, and X5 may be the same or different, and are independently selected from CR1 or N; Each R1 may be the same or different, and is independently selected from H, CN, halogen, unsubstituted, or optionally by one, two, or more Rs. c The following groups are substituted: OH, C 1-12 Alkyl, C 2-12 alkenyl, C 2-12 alkynyl, halogenated C 1-12 Alkyl, C 1-12 Alkoxy, C 3-12 Cycloalkyl, 3-14 membered heterocyclic groups, C 6-14 Aryl, 5-14 heteroaryl, -N(R) 11 (R) 12 -COR 13 , or -S(O)2R 14 ; Each R c They are either the same or different, and are independently selected from OH, -N(R) 15 (R) 16 CN, halogens, oxometalates (=O), C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 cycloalkyl or 3-6 membered heterocyclic groups; R 11 R 12 R 14 R 15 R 16 Whether the two are the same or different, they are selected independently from H and C. 1-6 Alkyl, Halogenated C 1-6 Alkyl or C 3-6 cycloalkyl; R 13 Selected from H, OH, -NH2, C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 cycloalkyl, 3-6 membered heterocyclic, C 6-10 Aryl or 5-10 heteroaryl groups; Ring A is a 5-10 member heteroaryl group; Each R a They may be identical or different, and are independently selected from CN, halogen, unsubstituted, or optionally composed of one, two, or more R. a1 The following groups are substituted: OH, C 1-12 Alkyl, Halogenated C 1-12 Alkyl, C 1-12 Alkoxy, C 1-12 Alkylthio, C 3-12 Cycloalkyl, 3-14 membered heterocyclic groups or -NH2; each R a1 They are selected independently of OH, -NH2, CN, halogens, oxo (=O), and C, whether they are the same or different. 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl or 3-6 membered heterocyclic groups; m is selected from 0, 1, 2, 3 or 4; Y is absent or selected from CO, CONR2, or unsubstituted or optionally substituted by one, two or more R3 groups, including 5-14 member heteroaryl groups or CONR2-C. 1-12 Alkylene; R2 is selected from H, C 1-6 Alkyl or C 3-6 Cycloalkyl; each R3 may be the same or different, and is independently selected from CN, halogen, OH, NH2, C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1- 6-alkoxy, C 3-6 Cycloalkyl or 3-6 membered heterocyclic groups; Ring B is a 3-14 member N-containing heterocyclic ring; Each R b They may be identical or different, and are independently selected from CN, halogen, oxo (=O), unsubstituted, or optionally substituted by one, two, or more R. b1 The following groups are substituted: OH, C 1-12 Alkyl, C 2-12 alkenyl, C 2-12 alkynyl, halogenated C 1-12 Alkyl, C 1- 12 Alkoxy, C 1-12 Alkylthio, C 3-12 Cycloalkyl, 3-14 membered heterocyclic groups, C 6-14 Aryl, 5-14 heteroaryl, -N(R) 41 (R) 42 ) or -S(O)2R 43 Or, two R atoms attached to the same carbon atom b Together with the carbon atom it is attached to, it forms an unsubstituted or optionally substituted form with one, two or more R atoms. b1 The following ring systems are replaced: C 3-14 A carbon ring or a 3-14 membered heterocycle; or, two R atoms attached to adjacent carbon atoms. b Together with the carbon atoms respectively attached thereto, they form unsubstituted or optionally substituted with one, two or more R atoms. b1 Substitution of the following groups: olefinic bond (C=C), C 3-14 Carbon rings, 3-14 membered heterocycles, C 6-14 Aromatic rings or 5-14 heterocyclic aromatic rings; R 41 R 42 R 43 Whether the two are the same or different, they are selected independently from H and C. 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 3-6 cycloalkyl; each R b1 They are selected independently of OH, -NH2, CN, halogens, oxo (=O), and C, whether they are the same or different. 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 1-6 Alkoxy, C 3-6 Cycloalkyl or 3-6 membered heterocyclic groups; n is selected from 0, 1, 2, 3, 4, or 5; and At least one of rings A and B is a bicyclic (such as a fused ring, a bridged ring, or a spiral ring) system.

2. The compound according to claim 1, its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt, or prodrug compound, characterized in that, X1 is selected from CR1 or N; X2, X3, X4, and X5 are CR1; Preferably, each R1 may be the same or different, and is independently selected from H, CN, halogens (e.g., F, Cl, Br), OH, C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy (e.g., OCF3) or C 3-6 cycloalkyl; Preferably, X1 is selected from CH, CF, or N; X2 is selected from CH, COCF3, or CCN; X3, X4, and X5 are CH. Preferably, Selected from Preferably, ring A is selected from Preferably, each R a Whether the same or different, they are independently selected from CN, halogen, and C. 1-6 Alkyl (e.g., methyl), halogenated C 1-6 Alkyl (e.g., CHF2), C 1-6 Alkoxy, halogenated C 1-6 Alkoxy or C 3-6 cycloalkyl; Preferably, m is selected from 0 or 1; Preferably, Selected from The "*" side is connected to Y, and the "#" side is connected to Y.

3. The compound according to claim 1 or 2, its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt, or prodrug compound, characterized in that, Y is absent or selected from CO, CONR2, CONR2-C(R3)2- or a 5-membered heteroaryl group (e.g., ...). )； Preferably, R2 is selected from H or C. 1-3 alkyl; Preferably, each R3 is identical or different, and is independently selected from H or C. 1-3 alkyl; Preferably, ring B is selected from Preferably, each R b They may be the same or different, and are independently selected from CN, halogen, oxo (=O), OH, and -NH. 2 -S(O)2C 1-3 Alkyl, C 1-6 Alkyl, Halogenated C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, C 3-6 Cycloalkyl, 3-6 membered heterocyclic, phenyl, or 5-6 membered heteroaryl; Preferably, the two R atoms attached to the same carbon atom b Together with the carbon atom it is attached to, it forms the following group: C 3-6 Carbon rings or 3-6 membered heterocycles; or, two R atoms attached to adjacent carbon atoms. b Together with the carbon atoms they are attached to, they form the following groups: olefinic bond (C=C), C 3-6 Carbon rings, 3-6 membered heterocycles, benzene rings, or 5-6 membered heteroaromatic rings; Preferably, n is 0, 1, or 2; Preferably, Selected from 4. The compound according to any one of claims 1-3, its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt, or prodrug compound, characterized in that, The compound shown in formula (I) has the following structure: in, X1 is selected from CR 1a Or N; R 1a Selected from H, halogens (e.g., F, Cl or Br), OCF3 or CN; X2 is CR 1b Or N; R 1b Selected from H, halogens (e.g., F, Cl or Br), OCF3 or CN; X6 is selected from CH or N; X7 is selected from O, S, or NH; R2 is selected from H or C. 1-3 alkyl; n is 0 or 2; When n is 2, two R atoms attached to the same carbon atom b Together with the carbon atom it is attached to, it forms the following group: C 3-6 Carbon rings or 3-6 membered heterocycles; or, two R atoms attached to adjacent carbon atoms. b Together with the carbon atoms they are attached to, they form the following groups: olefinic bond (C=C), C 3-6 Carbon rings, 3-6 membered heterocycles, benzene rings, or 5-6 membered heteroaromatic rings; q1 is selected from 1 or 2; q2 is selected from 0, 1 or 2.

5. The compound according to any one of claims 1-4, its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt, or prodrug compound, characterized in that, The compound shown in formula (I) has the following structure: The compound shown in formula (I) has the following structure: in, X1 is selected from CR 1a Or N; R 1a Selected from H, F, Cl, or Br; X2 is CR 1b ;R 1b Selected from H, OCF3, or CN; X7 is selected from O, S, or NH; R2 is selected from H or C. 1-3 alkyl; n is 2; and the two R atoms connected to the same carbon atom b Together with the carbon atom it is attached to, it forms the following group: C 3-6 Carbon rings or 3-6 membered heterocycles; or, two R atoms attached to adjacent carbon atoms. b Together with the carbon atoms they are attached to, they form the following groups: olefinic bond (C=C), C 3-6 Carbon rings, 3-6 membered heterocycles, benzene rings, or 5-6 membered heteroaromatic rings; q1 is selected from 1 or 2; q2 is selected from 0, 1, or 2; Preferably, the compound represented by formula (I) has the following structure: in, X1 is selected from CR 1a Or N; R 1a Selected from H, F, Cl, or Br; X2 is CR 1b ;R 1b Selected from H, OCF3, or CN; X7 is selected from O, S, or NH; R2 is selected from H or C. 1-3 alkyl; p is selected from 0 or 1; Each R3 may be the same or different, and is independently selected from H or C. 1-3 alkyl; n is selected from 0, 1, 2, or 3; Each R b They are the same or different, and are independently selected from halogens and C. 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkyl, Halogenated C 1-3 alkoxy group; or, two R groups attached to the same carbon atom. b Together with the carbon atom it is attached to, it forms the following group: C 3-6 Carbon rings or 3-6 membered heterocycles; or, two R atoms attached to adjacent carbon atoms. b Together with the carbon atoms they are attached to, they form the following groups: olefinic bond (C=C), C 3-6 Carbocyclic rings, 3-6 membered heterocyclic rings, benzene rings, or 5-6 membered heteroaromatic rings; or, R b R2, together with the atoms it is attached to, forms a 3-6 membered heterocycle; q1 is selected from 1 or 2; q2 is selected from 0, 1 or 2.

6. The compound according to any one of claims 1-5, its racemate, stereoisomer, tautomer, solvate, polymorph, pharmaceutically acceptable salt, or prodrug compound, characterized in that, The compound shown in formula (I) is selected from the following structures:

7. A method for preparing the compound of formula (I) according to any one of claims 1-6, comprising the following step A: Step A: in, X1, X2, X3, X4, X5, Ring A, Ring B, Y, R a R b m and n have the definitions of any one of claims 1-4.

8. A pharmaceutical composition comprising a therapeutically effective amount of at least one of the following: a compound of formula (I) according to any one of claims 1-6, a racemic mixture, a stereoisomer, a tautomer, a solvate, a polymorph, a pharmaceutically acceptable salt, or a prodrug compound thereof; Preferably, the pharmaceutical composition further includes one or more pharmaceutically acceptable excipients; Preferably, the pharmaceutical composition further contains one or more additional therapeutic agents.

9. A method of treating or preventing a disease or condition associated with mitochondrial dysfunction, comprising administering to a patient a preventive or therapeutically effective amount of at least one of the following: a compound of formula (I) as claimed in any one of claims 1-6, a racemic mixture, a stereoisomer, a tautomer, a solvate, a polymorph, a pharmaceutically acceptable salt thereof, or a prodrug compound of claim 8; Preferably, the diseases or conditions associated with mitochondrial dysfunction are Parkinson's disease, cancer and fibrosis, Leigh syndrome; multiple sclerosis; mitochondrial encephalopathy; mitochondrial encephalomyopathy with hyperlactatemia and stroke-like episodes (MELAS); Leber's hereditary optic neuropathy; ataxia; retinitis pigmentosa-maternally inherited Leigh syndrome; Danon's disease; diabetes; diabetic nephropathy; metabolic disorders; heart failure; ischemic heart disease leading to myocardial infarction; schizophrenia; polysulfatase deficiency; mucolipid storage disease II; mucolipid storage disease III; mucolipid storage disease IV; GMI-ganglioside storage disease; neuronal ceroid lipofuscin deposition disease; Alpes disease; Barth syndrome; β-oxidation deficiency; carnitine-acyl-carnitine deficiency; carnitine deficiency; creatine deficiency syndrome; coenzyme Q10 deficiency; complex I deficiency; complex II deficiency; complex III deficiency; complex IV deficiency; complex V deficiency; COX deficiency; chronic progressive lateral oculomotor palsy syndrome; CPT. I deficiency; CPT II deficiency; glutaric aciduria type II; Kiehl's syndrome; lactic acidosis; long-chain acyl-CoA dehydrogenase deficiency; Lewy body disease or syndrome; fatal infantile cardiomyopathy; Left disease; medium-chain acyl-CoA dehydrogenase deficiency; myoclonic epilepsy and fluffy red fibrinoid syndrome; mitochondrial cytopathic disease; mitochondrial degenerative ataxia syndrome; mitochondrial DNA depletion syndrome; musculoskeletal disorders and encephalopathy; Pearson syndrome; pyruvate dehydrogenase deficiency; pyruvate carboxylase deficiency; POLG mutation; medium / short 3-hydroxyacyl-CoA dehydrogenase deficiency; and very long-chain acyl-CoA dehydrogenase deficiency; and age-dependent decline in cognitive function and muscle strength; amyotrophic lateral sclerosis; Huntington's disease; focal ischemia; stroke; Lewy body dementia and frontotemporal dementia; Preferably, the patient includes a mammal, more preferably a human.

10. The use of at least one of the compounds of formula (I) according to any one of claims 1-6, their racemic mixtures, stereoisomers, tautomers, solvates, polymorphs, pharmaceutically acceptable salts or prodrug compounds, or the use of the pharmaceutical composition according to claim 8 in the preparation of a medicament; Preferably, the use is in the preparation of a medicament for the treatment or prevention of diseases or conditions associated with mitochondrial dysfunction; Preferably, the diseases or conditions associated with mitochondrial dysfunction are Parkinson's disease, cancer and fibrosis, Leigh syndrome; multiple sclerosis; mitochondrial encephalopathy; mitochondrial encephalomyopathy with hyperlactatemia and stroke-like episodes (MELAS); Leber's hereditary optic neuropathy; ataxia; retinitis pigmentosa-maternally inherited Leigh syndrome; Danon's disease; diabetes; diabetic nephropathy; metabolic disorders; heart failure; ischemic heart disease leading to myocardial infarction; schizophrenia; polysulfatase deficiency; mucolipid storage disease II; mucolipid storage disease III; mucolipid storage disease IV; GMI-ganglioside storage disease; neuronal ceroid lipofuscin deposition disease; Alpes disease; Barth syndrome; β-oxidation deficiency; carnitine-acyl-carnitine deficiency; carnitine deficiency; creatine deficiency syndrome; coenzyme Q10 deficiency; complex I deficiency; complex II deficiency; complex III deficiency; complex IV deficiency; complex V deficiency; COX deficiency; chronic progressive lateral oculomotor palsy syndrome; CPT. CPT II deficiency; glutaric aciduria type II; Kiehl's syndrome; lactic acidosis; long-chain acyl-CoA dehydrogenase deficiency; Lewy body disease or syndrome; fatal infantile cardiomyopathy; Left disease; medium-chain acyl-CoA dehydrogenase deficiency; myoclonic epilepsy and fluffy red fibrinoid syndrome; mitochondrial cytopathic disease; mitochondrial degenerative ataxia syndrome; mitochondrial DNA depletion syndrome; musculoskeletal disorders and encephalopathy; Pearson syndrome; pyruvate dehydrogenase deficiency; pyruvate carboxylase deficiency; POLG mutation; medium / short 3-hydroxyacyl-CoA dehydrogenase deficiency; and very long-chain acyl-CoA dehydrogenase deficiency; and age-dependent decline in cognitive function and muscle strength; amyotrophic lateral sclerosis; Huntington's disease; focal ischemia; stroke; Lewy body dementia and frontotemporal dementia.

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