Spiro derivatives as M4 activators / modulators and uses thereof

Abstract

The present disclosure provides a compound of Formula I: # imgabs0 # or an N-oxide thereof, or a pharmaceutically acceptable salt of the compound or the N-oxide, wherein: A, Y, m, n, p, R1, R2, R3, R3a, R4, R5, R6, R7, and Z are as described herein; the present disclosure provides processes for preparing such compounds, N-oxides, or salts; intermediates useful for the preparation of such compounds, N-oxides, or salts; the present invention relates to compounds, N-oxides, or salts, and compositions containing such compounds, N-oxides, or salts, and their use for the treatment of M4-mediated (or M4-related) disorders, including, for example, Alzheimer's disease, Parkinson's disease, schizophrenia (e.g., their cognitive and negative symptoms), pain, addiction, and sleep disorders.

Description

Technical Field

[0001] The present disclosure generally relates to novel pyridine azaspiro compounds that are agonists / activators / modulators of the muscarinic M4 receptor and are useful for treating M4-mediated diseases and disorders, including but not limited to schizophrenia, Alzheimer's disease, dementia-related psychoses, dementia with Lewy bodies, Parkinson's disease and related memory and executive dysfunction, bipolar disorder, agitation, and psychoses related thereto. Background Art

[0002] Patients suffering from schizophrenia, Alzheimer's disease, Parkinson's disease, Huntington's disease (Huntington's Disease), depression and various other neurological / neurodegenerative diseases often suffer from behavioral and cognitive impairment, resulting in debilitating disruptions to their daily lives. Over the years, many pharmacological treatments have been found to provide some improvements in behavioral and cognitive function. However, the improvements are modest at best, and it is often the case that the potential dose-limiting side effects (including extrapyramidal side effects and metabolic side effects) associated with these treatments result in partial responsiveness and non-compliance.

[0003] Muscarinic acetylcholine receptors (mAChRs) are a viable mechanism for treating these diseases. Five mAChR subtypes (M1-M5) have been identified, which are part of the G protein-coupled receptor (GPCR) superfamily. These subtypes are widely distributed in the periphery and central nervous system (CNS), with the M1 and M4 subtypes primarily expressed in the CNS. HTL0016878, an M4 agonist developed to treat the primary symptoms of Alzheimer's disease, has entered Phase 2 clinical studies.

[0004] Therefore, there is a need for agonists of the muscarinic M4 receptor to treat M4-mediated diseases and disorders, such as Parkinson's disease, schizophrenia, Alzheimer's disease, and other diseases and disorders described herein. Summary of the Invention

[0005] Provided herein are compounds having the structure of Formula (I): or its N-oxide, or a pharmaceutically acceptable salt of the compound or its N-oxide, wherein: A is a ring containing 1 or 2 nitrogen atoms and optionally 1 to 3 R A substituted 6-8 membered heterocyclic ring; each R A Independently C 1-3 Alkyl, halogen, =O, OH, C 1-3 Hydroxyalkyl or C 1-3Haloalkyl; Y is a bond, S, O, CH2, CHF, CF2 or C(OH)H; m is 1 or 2; n is 1 or 2; p is 1 or 2; R 1 H, halogen, CN, OH, NO2, -N(R 6 )(R 7 ), C 1-6 Alkyl, C 2-6 Heteroalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, -[O] 0-1 -C 3-6 Cycloalkyl, -[O] 0-1 -C 6-10 Aryl, -[O] 0-1 -4-8 membered heterocycle or -[O] 0-1 -5-10 membered heteroaryl, wherein the heterocycle and heteroaryl each contain 1, 2 or 3 ring heteroatoms selected from N, O and S, and when R 1 C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, -[O] 0-1 -C 3-6 Cycloalkyl, -[O] 0-1 -C 6-10 Aryl, -[O] 0-1 -4-8 membered heterocyclic ring, -[O] 0-1 -5-10 membered heteroaryl, -NH-C 3-6 Cycloalkyl, -NH-C 6-10 Aryl, -NH-4-8 membered heterocyclic ring, -NH-5-10 membered heteroaryl, -N(C 1-6 alkyl)-C 3-6 Cycloalkyl, -N(C 1-6 alkyl)-C 6-10 Aryl, -N(C 1-6 alkyl)-4-8 membered heterocyclic ring or -N(C 1-6 alkyl)-5-10 membered heteroaryl, R 1 0, 1, 2 or 3 independently selected from halogen, CN, OH, ═O, SO2 and C 1-3 Alkyl substituents substituted; R 2 H, halogen, CN, OH, –N(R 6 )(R 7 ), C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 2-6 Heteroalkyl, C3-6 Cycloalkyl or a 4-8 membered heterocycle containing 1, 2 or 3 ring heteroatoms selected from N, O and S; R 3 Halogen, C 0-6 Alkylene -CN, OH, -N(R 6 )(R 7 ), C 1-6 Alkyl, C 2-6 Heteroalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, C 1-6 Alkylene-OC 1-6 Alkyl, C 1-6 Alkylene-NHC(O)C 1-6 Alkyl, C 1-6 Alkylene-C(O)NHC 1-6 Alkyl, C 0-6 Alkylene-Cyc, OC 0-6 Alkylene-Cyc, NH-Cyc, N(C 1-6 alkyl)-Cyc or C(O)Cyc; Cyc is C 3-12 Cycloalkyl, C 3-6 Heterocycloalkyl, C 5-12 Spiroalkyl, C 5-12 Heterospirocycloalkyl, C 6-10 aryl, 3-12 membered heterocyclic ring or 5-10 membered heteroaryl, the heterocyclic ring or heteroaryl containing 1, 2 or 3 ring heteroatoms independently selected from N, O and S, and Cyc is replaced by 0, 1, 2 or 3 R 3a Substituent substitution; each R 3a independently selected from halogen, CN, OH, ═O, ═N(C 1-6 alkyl), SO2, C 1-6 Alkyl, C 2-10 Olefins, C 1-6 Hydroxyalkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 1-6 Alkylene-OC 1-6 Alkyl, C 0-6 Alkylene-C(O)C 1-6 Alkyl, C 0-6 Alkylene-NH2, C 0-6 Alkylene-NH(C 1-6 Alkyl), C 0-6 Alkylene-N(C 1-6 Alkyl)2, -SC 1-6 Alkyl, C 0-6 Alkylene-SO2C 1-6 Alkyl, C0-6 Alkylene-C(O)NH2, C 0-6 Alkylene-C(O)NH(C 1-6 Alkyl), C 0-6 Alkylene-C(O)N(C 1-6 Alkyl)2, C 0-6 Alkylene-NHC(O)C 1-6 Alkyl, C 0-6 Alkylene-COOH, C 0-6 Alkylene-CO2C 1-6 Alkyl, C 0-6 Alkylene-C 3-6 Cycloalkyl and C containing 1, 2 or 3 heteroatoms selected from N, O and S 0-6 Alkylene-3-6 membered heterocycle; R 4 H, halogen, CN, OH, C 1-6 Alkyl, C 1-6 Haloalkyl or C 1-6 Alkoxy; R 5 is -CO2-Z or its bioisostere; each R 6 and R 7 Independently H, C 1-6 Alkyl, C(O)-C 1-6 Alkyl, spiro or bicyclic C 8-14 cycloalkyl, an 8-14 membered heterocycle containing 1, 2 or 3 ring heteroatoms selected from N, O and S, and when R 6 or R 7 When not H, it is replaced by 0, 1, 2 or 3 independently selected from halogen, CN, ═O, SO 2 , OH, C 0-6 Alkylene-NH2, C 0-6 Alkylene-NH(C 1-6 Alkyl), C 0-6 Alkylene-N(C 1-6 Alkyl)2, C 0-6 Alkylene-SO2C 1-6 Alkyl, C 1-6 Alkyl and C 1-6 The alkoxy substituent is substituted, or

[0006] R 6 and R 7 Together with the nitrogen to which they are attached, they form a 4-10 membered heterocyclic ring containing 0-2 additional ring heteroatoms independently selected from N, O and S; and Z is C 1-7 Alkyl, C 1-7 Halogenated alkyl, C 3-6 Cycloalkyl or C 2-6 Alkyne, and Z is replaced by 0, 1, 2 or 3 C 1-6 Alkoxy or C 3-6Cycloalkyl substitution; provided that when R 1 and R 4 Each is H, m and p are each 1, and A is And (a) R 2 is H, Y is CH2, n is 1, and R 3 CH3, OCH3, or (b) R 2 is F, Y is CH2, n is 1, and R 3 for or (c)R 2 is H, Y is a bond, n is 1, and R 3 for or (d)R 2 is H, Y is a bond, n is 2, and R 3 for When R 5 is not CO2CH2CH3 or CO2CH(CH3)3. In some embodiments, one or more H, including but not limited to any R 1 、R 2 、R 3 and / or R 4 Those present in the group may be replaced by D.

[0007] Also provided herein are pharmaceutical compositions comprising compounds as disclosed herein. Also provided are methods of treating an M4-mediated (or M4-related) disease or disorder associated with abnormal M4 receptor activity in a subject, comprising administering to the subject a therapeutically effective amount of a compound as disclosed herein.

[0008] By reviewing the following detailed description in conjunction with the accompanying drawings, other aspects and advantages will be apparent to those skilled in the art. Although the compounds and methods disclosed herein are susceptible to various forms of embodiments, the following description includes specific embodiments, it should be understood that this disclosure is illustrative and is not intended to limit the invention to the specific embodiments described herein. DETAILED DESCRIPTION

[0009] Provided herein are compounds having the structure of Formula (I):

[0010] Or a pharmaceutically acceptable salt thereof. Also provided herein are compounds that act on the M4 receptor. The compounds described herein can be used to treat M4-mediated diseases or M4-related diseases.

[0011] Chemical definition

[0012] As used herein, the term "alkyl" refers to straight-chain and branched saturated hydrocarbon groups containing 1 to 30 carbon atoms (e.g., 1 to 20 carbon atoms or 1 to 10 carbon atoms). n It means that the alkyl group has "n" carbon atoms. For example, C6 alkyl refers to an alkyl group with 6 carbon atoms. 1-7 Alkyl refers to an alkyl group having a carbon number covering the entire range (i.e., 1 to 7 carbon atoms) and all subgroups (e.g., 2-6, 2-5, 3-6, 2, 3, 4, 5, 6, and 7 carbon atoms). Non-limiting examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl (2-methylpropyl), and tert-butyl (1,1-dimethylethyl). Unless otherwise specified, an alkyl group may be an unsubstituted alkyl group or a substituted alkyl group.

[0013] As used herein, the term "alkylene" refers to a divalent saturated aliphatic group. n It means an alkylene group having "n" carbon atoms, for example, C1 alkylene is CH2. 1-6 Alkylene refers to an alkylene group having a number of carbon atoms encompassing the entire range and all subgroups as previously described for "alkyl" groups. Co alkylene represents a direct bond (i.e., no alkylene linker).

[0014] As used herein, the term "alkene" or "alkenyl" is defined the same as "alkyl," except that it contains at least one carbon-carbon double bond and has 2 to 30 carbon atoms, such as 2 to 20 carbon atoms or 2 to 10 carbon atoms. n It means that the alkenyl group has "n" carbon atoms. For example, C4 alkenyl refers to an alkenyl group with 4 carbon atoms. 2-7 Alkenyl refers to an alkenyl group having a carbon number encompassing the entire range (i.e., 2 to 7 carbon atoms) and all subgroups (e.g., 2-6, 2-5, 3-6, 2, 3, 4, 5, 6, and 7 carbon atoms). Specifically contemplated alkenyl groups include ethenyl, 1-propenyl, 2-propenyl, and butenyl. Unless otherwise indicated, an alkenyl group may be an unsubstituted alkenyl group or a substituted alkenyl group. Unless otherwise indicated, an alkenyl group may be a cis-alkenyl group or a trans-alkenyl group.

[0015] As used herein, the term "alkyne" or "alkynyl" is defined the same as "alkyl," except that it contains at least one carbon-carbon triple bond and has 2 to 30 carbon atoms, such as 2 to 20 carbon atoms or 2 to 10 carbon atoms. n It means that the alkynyl group has "n" carbon atoms. For example, C4 alkynyl refers to an alkynyl group with 4 carbon atoms. 2-7Alkynyl refers to an alkynyl group having a number of carbon atoms encompassing the entire range (i.e., 2 to 7 carbon atoms) as well as all subgroups (e.g., 2-6, 2-5, 3-6, 2, 3, 4, 5, 6, and 7 carbon atoms). Specifically contemplated alkynyl groups include ethynyl, 1-propynyl, 2-propynyl, and butynyl. Unless otherwise specified, an alkynyl group can be unsubstituted or substituted.

[0016] As used herein, the term "cycloalkyl" specifically refers to a non-aromatic ring in which each atom of the ring is carbon, i.e., a carbocycle, and may be monocyclic, bicyclic, bridged, fused, or spirocyclic. n It means that the cycloalkyl group has "n" ring carbon atoms. For example, C5 cycloalkyl refers to a cycloalkyl group having 5 ring carbon atoms in the ring. 3-12 Cycloalkyl refers to cycloalkyl groups having ring carbon number encompassing the entire range (i.e., 3 to 12 carbon atoms) and all subgroups (e.g., 4-8, 3-7, 4-7, 3-6, 4-6, 3-5, 4-5, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 12 carbon atoms). Non-limiting examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Non-limiting examples of bridged cycloalkyl groups include Non-limiting examples of spirocycloalkyl groups include Non-limiting examples of fused cycloalkyl groups are (For example, ). Unless otherwise specified, a cycloalkyl group may be an unsubstituted cycloalkyl group or a substituted cycloalkyl group.

[0017] As used herein, the term "heterocycle" is similar to the definition of cycloalkyl, except that the ring contains 1 to 4 heteroatoms independently selected from oxygen, nitrogen and sulfur. In addition, the heterocycle of the present disclosure can be monocyclic, bicyclic, bridged, fused or spirocyclic. For example, the heterocycle can be a 3-12 membered ring having 1 or 2 or 3 heteroatoms selected from N, O and S, a monocyclic, bicyclic, bridged, fused or spirocyclic ring. For another example, the heterocycle can be a 5-12 membered or 8-10 membered bicyclic, bridged, fused or spirocyclic group having 1 or 2 or 3 ring heteroatoms selected from N, O and S in the bicyclic ring. Non-limiting examples of heterocyclic groups include piperidine, piperazine, tetrahydrofuran, tetrahydropyran, dihydrofuran, morpholine, oxazepane, thiazole, pyrrole and pyridine. Non-limiting examples of heterocyclic groups include

[0018] The cycloalkyl and heterocyclic groups may be saturated or partially unsaturated ring systems (e.g., with double or triple bonds), but these groups are not aromatic. The cycloalkyl and heterocyclic groups may be optionally substituted, for example, by 1 to 3 groups independently selected from the group consisting of alkyl, alkoxy, alkylene, OH, C 0-6 Alkylene-C(O)NH2, NH2, =O, SO2, aryl, haloalkyl, haloalkoxy, C 0-6 Alkylene-C(O)-alkyl, C 0-6 Alkylene-SO2 alkyl, halogen, OH, NHC 1-3 Alkylene-aryl, OC 1-3 Alkylene-aryl, C 1-3 Alkylene-aryl and C having 1-3 heteroatoms selected from N, O and S 0-6 Alkylene-C 3-6 Other contemplated substitutions are discussed in detail elsewhere in this disclosure.

[0019] As used herein, the term "aryl" refers to an aromatic ring in which each atom of the ring is carbon and can be a monocyclic or polycyclic (e.g., fused bicyclic and fused tricyclic) carbocyclic aromatic ring system. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, tetrahydronaphthyl, phenanthrenyl, biphenylene, indanyl, indenyl, anthracenyl, fluorenyl, tetrahydronaphthyl. Unless otherwise indicated, an aryl group can be an unsubstituted aryl group or a substituted aryl group.

[0020] As used herein, the term "heteroaryl" refers to an aromatic heterocycle, and may be a monocyclic or polycyclic (e.g., fused bicyclic and fused tricyclic) aromatic ring system in which one to four (e.g., one to three) ring atoms are selected from oxygen, nitrogen, and sulfur, and the remaining ring atoms are carbon, the ring system being attached to the remainder of the molecule via any ring atom. Non-limiting examples of heteroaryl groups include, but are not limited to, pyridyl, pyridazinyl, pyrazinyl, pyrimidinyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiadiazolyl, oxadiazolyl, furanyl, thienyl, quinolyl, isoquinolyl, benzoxazolyl, benzimidazolyl, benzofuranyl, benzothiazolyl, triazinyl, triazolyl, purinyl, pyrazinyl, purinyl, indolinyl, phthalazinyl, indazolyl, quinolyl, isoquinolyl, cinnolinyl, quinazolinyl, naphthyridinyl, pyridopyridinyl, indolyl, 3H-indolyl, pteridinyl, and quinoxalinyl. Non-limiting examples of heteroaryl groups include Unless otherwise specified, a heteroaryl group may be an unsubstituted heteroaryl group or a substituted heteroaryl group.

[0021] As used herein, the term "hydroxy" or "hydroxyl" refers to a "-OH" group. Thus, a "hydroxyalkyl" group refers to an alkyl group substituted with one or more -OH groups.

[0022] As used herein, the term "alkoxy" or "alkoxyl" refers to an "-O-alkyl" group.

[0023] As used herein, the term "halogen" is defined as fluorine, chlorine, bromine and iodine. Therefore, "haloalkyl" refers to an alkyl group substituted with one or more halogen atoms. In some cases, the haloalkyl group is a perhaloalkyl, i.e., all hydrogen atoms of the alkyl group have been replaced with halogen. Some non-limiting examples of haloalkyl groups include CF , CHF , CH F, CCl , CH Cl and CH CF . Similarly, "haloalkoxy" refers to an alkoxy group substituted with one or more halogen atoms, such as OCF . And "haloalkyl" refers to an alkyl group in which one or more hydrogen atoms have been replaced with one or more halogens.

[0024] As used herein, the term "heteroalkyl" refers to an alkyl chain interrupted by one or more heteroatoms selected from N, O, and S and having 2 to 30 carbon atoms (e.g., 2 to 20 carbon atoms or 2 to 10 carbon atoms). n It means that the heteroalkyl group has "n" carbon atoms. For example, C6 heteroalkyl refers to an alkyl group with 6 carbon atoms and the carbon chain is interrupted by one or more heteroatoms. 2-6 Heteroalkyl refers to an alkyl group having a number of carbon atoms encompassing the entire range (i.e., 2 to 6 carbon atoms) as well as all subgroups (e.g., 2-5, 3-6, 3-5, 4-6, 2, 3, 4, 5, and 6 carbon atoms). The number of heteroatoms in the heteroalkyl chain can be, for example, 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, or 1 or 2. Specific numbers of heteroatoms contemplated include 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10 heteroatoms.

[0025] As used herein, the term "bioisostere" refers to a molecule resulting from the exchange of one atom or group of atoms for another or another broadly similar group of atoms. For example, an ester group can be replaced by one of the following bioisosteres of an ester, including but not limited to acylsulfonamide (CO—NRSO2R), hydroxamic acid (CONROH), hydroxamate (CONROR), tetrazole, hydroxyisoxazole, isoxazol-3-one, and sulfonamide (SO2NR2), where each R can independently represent hydrogen, alkyl, fluoroalkyl, carbocyclyl, carbocyclylalkyl, aryl, aralkyl, heterocycloalkyl, heterocycloalkylalkyl, heteroaryl, or heteroarylalkyl.

[0026] A "substituted" functional group (e.g., a substituted alkyl, cycloalkyl, aryl, or heteroaryl group) is a functional group having at least one hydrogen group replaced by a non-hydrogen group (i.e., a substituent). Examples of non-hydrogen groups (or substituents) include, but are not limited to, alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, ether, aryl, O-alkylenearyl, N-alkylenearyl, alkylenearyl, heteroaryl, heterocycle, hydroxyl, hydroxyalkyl, haloalkoxy, amido, =0, SO2, alkoxy, ester, thioester, acyl, carboxyl, cyano, nitro, amino, thiol, and halogen. When a substituted alkyl group includes more than one non-hydrogen group, the substituents may be attached to the same carbon or to two or more different carbon atoms.

[0027] Compounds of the present disclosure

[0028] Disclosed herein are compounds having the structure of Formula (I):

[0029]

[0030] or an N-oxide thereof, or a pharmaceutically acceptable salt of the compound or its N-oxide, wherein:

[0031] A is a ring containing 1 or 2 nitrogen atoms and optionally 1 to 3 R A substituted 6-8 membered heterocycle;

[0032] Each R A Independently C 1-3 Alkyl, halogen, =O, OH, C 1-3 Hydroxyalkyl or C 1-3 alkyl halide;

[0033] Y is a bond, S, O, CH2, CHF, CF2 or C(OH)H;

[0034] m is 1 or 2;

[0035] n is 1 or 2;

[0036] p is 1 or 2;

[0037] R 1 H, halogen, CN, OH, NO2, -N(R 6 )(R 7 ), C 1-6 Alkyl, C 2-6 Heteroalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, -[O] 0-1 -C 3-6 Cycloalkyl, -[O] 0-1 -C 6-10 Aryl, -[O] 0-1 -4-8 membered heterocycle or -[O] 0-1 -5-10 membered heteroaryl, wherein the heterocycle and heteroaryl each contain 1, 2 or 3 ring heteroatoms selected from N, O and S, and when R 1 C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, -[O] 0-1 -C 3-6 Cycloalkyl, -[O] 0-1 -C 6-10 Aryl, -[O] 0-1 -4-8 membered heterocyclic ring, -[O] 0-1 -5-10 membered heteroaryl, -NH-C 3-6 Cycloalkyl, -NH-C 6-10 Aryl, -NH-4-8 membered heterocyclic ring, -NH-5-10 membered heteroaryl, -N(C 1-6 alkyl)-C 3-6 Cycloalkyl, -N(C 1-6 alkyl)-C 6-10 Aryl, -N(C 1-6 alkyl)-4-8 membered heterocyclic ring or -N(C 1-6 alkyl)-5-10 membered heteroaryl, R 1 0, 1, 2 or 3 independently selected from halogen, CN, OH, ═O, SO2 and C 1-3 Alkyl substituents are substituted,

[0038] R 2 H, halogen, CN, OH, –N(R 6 )(R 7 ), C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 2-6 Heteroalkyl, C 3-6cycloalkyl or a 4-8 membered heterocycle containing 1, 2 or 3 ring heteroatoms selected from N, O and S;

[0039] R 3 Halogen, C 0-6 Alkylene -CN, OH, -N(R 6 )(R 7 ), C 1-6 Alkyl, C 2-6 Heteroalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, C 1-6 Alkylene-OC 1-6 Alkyl, C 1-6 Alkylene-NHC(O)C 1-6 Alkyl, C 1-6 Alkylene-C(O)NHC 1-6 Alkyl, C 0-6 Alkylene-Cyc, OC 0-6 Alkylene-Cyc, NH-Cyc, N(C 1-6 alkyl)-Cyc or C(O)Cyc;

[0040] Cyc is C 3-12 Cycloalkyl, C 3-6 Heterocycloalkyl, C 5-12 Spiroalkyl, C 5-12 Heterospirocycloalkyl, C 6-10 aryl, 4-12 membered heterocyclic or 5-10 membered heteroaryl, the heterocyclic or heteroaryl containing 1, 2 or 3 ring heteroatoms independently selected from N, O and S, and Cyc is replaced by 0, 1, 2 or 3 R 3a Substituent substitution;

[0041] Each R 3a independently selected from halogen, CN, OH, ═O, ═N(C 1-6 alkyl), SO2, C 1-6 Alkyl, C 2-10 Olefins, C 1-6 Hydroxyalkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 1-6 Alkylene-OC 1-6 Alkyl, C 0-6 Alkylene-C(O)C 1-6 Alkyl, C 0-6 Alkylene-NH2, C 0-6 Alkylene-NH(C 1-6 Alkyl), C 0-6 Alkylene-N(C 1-6Alkyl)2, -SC 1-6 Alkyl, C 0-6 Alkylene-SO2C 1-6 Alkyl, C 0-6 Alkylene-C(O)NH2, C 0-6 Alkylene-C(O)NH(C 1-6 Alkyl), C 0-6 Alkylene-C(O)N(C 1-6 Alkyl)2, C 0-6 Alkylene-NHC(O)C 1-6 Alkyl, C 0-6 Alkylene-COOH, C 0-6 Alkylene-CO2C 1-6 Alkyl, C 0-6 Alkylene-C 3-6 Cycloalkyl and C containing 1, 2 or 3 heteroatoms selected from N, O and S 0-6 Alkylene-3-6 membered heterocycle;

[0042] R 4 H, halogen, CN, C 1-6 Alkyl, OH or C 1-6 alkoxy;

[0043] R 5 is -CO2-Z or its bioisostere;

[0044] Each R 6 and R 7 Independently H, C 1-6 Alkyl, C(O)-C 1-6 Alkyl, spiro or bicyclic C 8-14 cycloalkyl, an 8-14 membered heterocycle containing 1, 2 or 3 ring heteroatoms selected from N, O and S, and when R 6 or R 7 When not H, it is replaced by 0, 1, 2 or 3 independently selected from halogen, CN, ═O, SO 2 , OH, C 0-6 Alkylene-NH2, C 0-6 Alkylene-NH(C 1-6 Alkyl), C 0-6 Alkylene-N(C 1-6 Alkyl)2, C 0-6 Alkylene-SO2C 1-6 Alkyl, C 1-6 Alkyl and C 1-6 The alkoxy substituent is substituted, or

[0045] R 6 and R 7together with the nitrogen to which they are attached, form a 4-10 membered heterocyclic ring containing 0-2 additional ring heteroatoms independently selected from N, O and S; and

[0046] Z is C 1-7 Alkyl, C 1-7 Halogenated alkyl, C 3-6 Cycloalkyl or C 2-6 Alkyne, and Z is replaced by 0, 1, 2 or 3 C 1-6 Alkoxy or C 3-6 cycloalkyl substitution;

[0047] The condition is that when R 1 and R 4 Each is H, m and p are each 1, and A is and

[0048] (a)R 2 is H, Y is CH2, n is 1, and R 3 CH3, OCH3, or

[0049] (b)R 2 is F, Y is CH2, n is 1, and R 3 for or

[0050] (c)R 2 is H, Y is a bond, n is 1, and R 3 for or

[0051] (d)R 2 is H, Y is a bond, n is 2, and R 3 for hour,

[0052] Then R 5 is not CO2CH2CH3 or CO2CH(CH3)3. In some embodiments, one or more H, including but not limited to any R 1 、R 2 、R 3 and / or R 4 Those present in the group may be replaced by D.

[0053] In each case, the ring containing the Y substituent Optionally, C 1-3 Alkyl, halogen, C 1-3 In some cases, the ring containing the Y substituent is substituted with methyl, fluoro, CF 3 , OH or CN.

[0054] In each case, A is and X is N, CH, C(OH) or CF.

[0055] In some cases, the compound has the structure of Formula (Ia):

[0056] In various cases, Y is CH2, CHF, CF2, or C(OH)H. In some cases, Y is CH2. In some cases, m is 1. In some cases, n is 1. In some cases, p is 1.

[0057] In each case, the compound has the structure of Formula (Ib): In each case, the compound has the structure of Formula (Ic): In each case, the compound has the structure of Formula (Id): In each case, the compound has the structure of Formula (Ie):

[0058] In some cases, R 5 is a CO2Z bioisostere and is selected from

[0059] In each case, R 5 Selected from CO2C 1-7 alkyl, In some cases, R 5 It is CO2CH2CH3.

[0060] In each case, R 1 H, halogen, CN, OH, -N(R 6 )(R 7 ), C 1-6 Alkyl or C 1-6 In some cases, R 1 is H or halogen.

[0061] In each case, R 2 H, halogen, CN, OH, -N(R 6 )(R 7 ), C 1-6Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy or C 1-6 In some cases, wherein R 2 H, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl or C 1-6 In some cases, R 2 is H or halogen.

[0062] In each case, each R 6 and R 7 Independently H, C 1-6 Alkyl or C(O)-C 1-6 In some cases, each R 6 and R 7 are independently H or C 1-6 In some cases, at least one R 6 and R 7 Together with the nitrogen to which they are attached they form a 4-10 membered heterocyclic ring containing 0-2 additional ring heteroatoms independently selected from N and O.

[0063] In each case, R 4 is H or halogen.

[0064] In each case, R 1 、R 2 and R 4 At least one of is halogen. In some cases, R 1 、R 2 and R 4 At least one of them is F.

[0065] In each case, R 3 C 0-6 Alkylene-Cyc, OC 0-6 Alkylene-Cyc, NH-Cyc, N(C 1-6 alkyl)-Cyc or C(O)Cyc, wherein Cyc is C 3-6 Cycloalkyl, C 6-10 aryl, 4-8 membered heterocyclic or 5-10 membered heteroaryl, the heterocyclic or heteroaryl containing 1, 2 or 3 ring heteroatoms independently selected from N, O and S, and Cyc is replaced by 0, 1, 2 or 3 R 3a In each case, R 3 C 0-6 Alkylene-Cyc, OC 0-6 Alkylene-Cyc, NH-Cyc, N(C 1-6alkyl)-Cyc or C(O)Cyc, wherein Cyc is C 3-12 Cycloalkyl, C 6-10 aryl, 4-12 membered heterocyclic or 5-10 membered heteroaryl, the heterocyclic or heteroaryl containing 1, 2 or 3 ring heteroatoms independently selected from N, O and S, and Cyc is replaced by 0, 1, 2 or 3 R 3a In each case, R 3 For -[O] 0-1 -C 3-6 Cycloalkyl, -[O] 0-1 -C 6-10 Aryl, -[O] 0-1 -4-8 membered heterocycle or -[O] 0-1 -5-10 membered heteroaryl, and optionally substituted by 1, 2 or 3 R 3a In some cases, R 3 for and 0, 1, 2 or 3 R 3a In each case, R 3 for and 0, 1, 2 or 3 R 3a In some cases, R 3 is unsubstituted. In each case, R 3 By 1 or 2 R 3a In some cases, R 3 By 1 R 3a In some cases, at least one R 3a Halogen, CN, OH, =O, SO2, C 1-6 Alkyl, C 2-10 Olefins, C 1-6 Hydroxyalkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 1-6 Alkylene-OC 1-6 Alkyl, N(C 1-6 Alkyl)2, C 1-6 Alkylene-N(C 1-6 Alkyl)2, -SC 1-6 Alkyl, NHC(O)C 1-6 Alkyl, C 1-6 Alkylene-NHC(O)C 1-6 an alkyl group or a C0-6 Alkylene-3-6 membered heterocycle. In some cases, at least one R 3a For CH3, CH2CH3, CH(CH3)2, CH2CH(CH3)2, CF3, CH2CH2F, CH2CHF2, CH2OH, C(CH3)2OH, CH2OCH3, CH2CH2OCH3, CH2O CH2CH3, F, CN, =O, SO2, OH, OCH3, OCH2CH3, OCH(CH3)2, OCHF2, CH2OCH3, CH2OCF3, SCH3, N(CH3)2, NHCOCH3, CD3, In each case, at least one R 3a is CH3, CH2CH3, F, CN, OH, OCH3, CF3, CH2OH or OCHF2.

[0066] Some specific examples of the compounds disclosed herein (or pharmaceutically acceptable salts thereof) are shown in Table A.

[0067] Table A

[0068]

[0069]

[0070]

[0071]

[0072]

[0073]

[0074]

[0075]

[0076]

[0077]

[0078]

[0079]

[0080]

[0081]

[0082]

[0083]

[0084]

[0085]

[0086]

[0087]

[0088]

[0089]

[0090]

[0091]

[0092]

[0093]

[0094]

[0095]

[0096]

[0097]

[0098]

[0099]

[0100]

[0101]

[0102]

[0103]

[0104]

[0105]

[0106]

[0107]

[0108]

[0109]

[0110]

[0111]

[0112]

[0113]

[0114]

[0115]

[0116]

[0117]

[0118]

[0119]

[0120]

[0121]

[0122]

[0123]

[0124]

[0125]

[0126] The present disclosure also includes all pharmaceutically acceptable isotopically labeled compounds identical to those recited herein, wherein one or more atoms are replaced by an atom having the same atomic number but an atomic mass or mass number different from the atomic mass or mass number prevalent in nature. Examples of suitable isotopes for inclusion in the compounds of the present disclosure include, but are not limited to, isotopes of hydrogen, such as 2 H. 3 H; isotopes of carbon, such as 11 C. 13 C and 14 C; isotopes of chlorine, such as 36 Cl; isotopes of fluorine, such as 18 F; iodine, such as 123 I and 125 I; nitrogen, such as 13N and 15 N; isotopes of oxygen, such as 15 O. 17 O and 18 O; isotopes of phosphorus, such as 32 P; and isotopes of sulfur, such as 35 S. Isotopically labeled compounds may include combinations of two or more of the same or different isotopes described above. Certain isotopically labeled compounds of the present disclosure, such as those incorporating radioactive isotopes, are useful in drug and / or substrate tissue distribution studies (e.g., assays). The radioactive isotope tritium, i.e. 3 H or "T", and carbon-14, i.e. 14 C, are particularly useful for this purpose in view of their ease of incorporation and facile means of detection. 2 H or "D")) substitution may offer certain therapeutic advantages resulting from greater metabolic stability (e.g., increased in vivo half-life or reduced dosage requirements) and therefore may be preferred in some circumstances. 11 C. 18 F. 15 O and 13 N) substitution can be used in positron emission tomography (PET) studies to examine substrate receptor occupancy. Isotope-labeled compounds of the present disclosure can generally be prepared by conventional techniques known to those skilled in the art or by methods similar to those described in the accompanying schemes and / or examples and preparations, using appropriate isotope-labeled reagents instead of previously employed unlabeled reagents. Pharmaceutically acceptable solvates according to the present disclosure include those in which the crystallization solvent can be isotopically substituted, such as D2O, acetone-d6 or DMSO-d6.

[0127] Has one or more keys with dashed lines and bold wedges (i.e. and ) are intended to indicate the absolute stereochemistry of the stereocenters present in the chemical structure. Bonds represented by simple lines do not indicate stereo preference. Bonds represented by dashed or bold straight lines (i.e. and ) are intended to indicate the relative stereochemistry of stereocenters present in a chemical structure. Unless otherwise indicated to the contrary, chemical structures that include one or more stereocenters exemplified herein but for which absolute or relative stereochemistry is not indicated encompass all possible stereoisomeric forms (e.g., diastereomers, enantiomers) of the compound and mixtures thereof. Structures with a single bold or wedge-shaped dashed line and at least one additional simple line encompass a single enantiomeric series of all possible diastereomers. Similarly, chemical structures with alkenyl groups are intended to encompass both cis and trans orientations, or, when substituted, encompass E-isomers and Z-isomers of the chemical structure.

[0128] Pharmaceutically acceptable salts and co-crystals

[0129] As used herein, the phrase "pharmaceutically acceptable salt" refers to salts of a compound that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and lower animals without undue adverse effects such as toxicity, irritation, allergic response, and the like, and are commensurate with a reasonable benefit / risk ratio.

[0130] Pharmaceutically acceptable salts are well known in the art. For example, SM Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, incorporated herein by reference. Pharmaceutically acceptable salts of the compounds described herein include those derived from suitable inorganic and organic acids and bases. These salts can be prepared in situ during the final isolation and purification of the compound.

[0131] As used herein, "Formula (I)", "Formula (la)", "Formula (lb)", "Formula (Ic)", "Formula (Id)" and "Formula (le)" are also defined to include all forms of the compounds of the present disclosure, including but not limited to their hydrates, solvates, isomers (including, for example, rotational stereoisomers), crystalline forms and non-crystalline forms, isomorphs, polymorphs, metabolites, prodrugs. For example, the compounds disclosed herein, or pharmaceutically acceptable salts thereof, may exist in unsolvated forms and solvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like. When the solvent or water is tightly bound, the complex will have a well-defined stoichiometry that is independent of humidity. However, when the solvent or water is weakly bound, as in channel solvates and hygroscopic compounds, the water / solvent content will depend on humidity and drying conditions. In such cases, non-stoichiometry will be the standard. In general, for the purposes of this disclosure, the solvated forms are considered equivalent to the unsolvated forms.

[0132] The compounds of the present disclosure may exist as inclusion compounds or other complexes (e.g., co-crystals). Included within the scope of the present disclosure are complexes such as inclusion compounds, drug-host inclusion complexes, wherein the drug and host are present in stoichiometric or non-stoichiometric amounts. Also included are complexes of the compounds of the present disclosure containing two or more organic and / or inorganic components, which may be in stoichiometric or non-stoichiometric amounts. The resulting complex may be ionized, partially ionized, or non-ionized. For a review of such complexes, see J. Pharm. Sci., 64 (8), 1269-1288, Haleblian (August 1975). Co-crystals are generally defined as crystalline complexes of neutral molecular components bound together by non-covalent interactions, but may also be complexes of neutral molecules and salts. Co-crystals can be prepared by melt crystallization, by recrystallization from a solvent, or by physically grinding the components together; see O. Almarsson and MJ Zaworotko, Chem. Commun. 2004, 17, 1889-1896. For a general review of multicomponent complexes, see JK Haleblian, J. Pharm. Sci. 1975, 64, 1269-1288.

[0133] In some cases, the compounds of the present disclosure can exist as atropisomers (e.g., one or more atropisomers) and / or be separated as atropisomers. Those skilled in the art will recognize that atropisomerism can exist in compounds having two or more aromatic rings (e.g., two aromatic rings connected by a single bond). See, for example, Freedman, TB et al., Absolute Configuration Determination of Chiral Molecules in the Solution State Using Vibrational Circular Dichroism. Chirality 2003, 15, 743-758; and Bringmann, G. et al., Atroposelective Synthesis of Axially Chiral Biaryl Compounds. Angew. Chem., Int. Ed. 2005, 44, 5384-5427.

[0134] When any racemate crystallizes, two different types of crystals are possible. The first type is the racemic compound (true racemate) mentioned above, in which one homogeneous form of crystal is produced containing equimolar amounts of both enantiomers. The second type is a racemic mixture or aggregate, in which both forms of crystal are produced in equimolar amounts, each form containing a single enantiomer.

[0135] The compounds of the present disclosure may also exist as an N-oxide or a pharmaceutically acceptable salt of the compound or N-oxide.

[0136] As known to those skilled in the art, amine compounds (i.e., those containing one or more nitrogen atoms), such as tertiary amines, can form N-oxides (also known as amine oxides or amine N-oxides). Typically, N-oxides have the formula R3N + -O~, wherein the parent amine R3N can be, for example, a tertiary amine (e.g., each R is independently alkyl, arylalkyl, aryl, heteroaryl, etc.), a heterocyclic amine, or a heteroaromatic amine (e.g., R3N together form 1-alkylpiperidine, 1-alkylpyrrolidine, 1-benzylpyrrolidine, or pyridine). For example, an imine nitrogen, particularly a heterocyclic imine nitrogen or a heteroaromatic imine nitrogen, or a pyridinic nitrogen (=N-) atom, such as a nitrogen atom in pyridine, pyridazine, or pyrazine, can be N-oxidized to form a ≡N-containing + -O – Thus, compounds according to the present disclosure that contain one or more nitrogen atoms (e.g., imine nitrogen atoms) may be capable of forming N-oxides thereof (e.g., mono-N-oxides, bis-N-oxides, or poly-N-oxides, or mixtures thereof, depending on the number of nitrogen atoms that are suitable for forming stable N-oxides).

[0137] As used herein, the term "N-oxide" refers to all possible and in particular all stable N-oxide forms of the amine compounds described herein (e.g., compounds containing one or more imine nitrogen atoms), such as mono-N-oxides (including different isomers when more than one nitrogen atom of the amine compound can form a mono-N-oxide) or poly-N-oxides (e.g., bis-N-oxides), or mixtures thereof in any ratio.

[0138] As mentioned above, compound of the present disclosure (or its N-oxide) can exist in the form of a pharmaceutically acceptable salt derived from an inorganic acid or an organic acid. Depending on the specific compound, due to one or more physical properties of the salt, such as enhanced drug stability under different temperatures and humidity, or desired solubility in water or oil, the salt of the compound may be advantageous. In some cases, the salt of the compound can also be used as an auxiliary agent in the separation, purification and / or splitting of the compound.

[0139] When the salt is intended for administration to a patient (as opposed to, for example, use in an in vitro setting), the salt is preferably pharmaceutically acceptable. The term "pharmaceutically acceptable salt" refers to a salt prepared by combining a compound of the present disclosure with an acid whose anion is generally considered suitable for human consumption or a base whose cation is generally considered suitable for human consumption. Pharmaceutically acceptable salts are particularly useful as products of the methods of the present disclosure because they have a higher water solubility relative to the parent compound.

[0140] Where applicable, suitable pharmaceutically acceptable acid addition salts of the disclosed compounds include those derived from inorganic and organic acids such as, but not limited to, hydrochloric acid, hydrobromic acid, hydrofluoric acid, boric acid, fluoroboric acid, phosphoric acid, metaphosphoric acid, nitric acid, carbonic acid, sulfonic acid, and sulfuric acid, and organic acids such as acetic acid, benzenesulfonic acid, benzoic acid, citric acid, ethanesulfonic acid, fumaric acid, gluconic acid, glycolic acid, isethionic acid, lactic acid, lactobionic acid, maleic acid, malic acid, methanesulfonic acid, trifluoromethanesulfonic acid, succinic acid, toluenesulfonic acid, tartaric acid, and trifluoroacetic acid. Suitable organic acids generally include, but are not limited to, aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, carboxylic, and sulfonic organic acids.

[0141] Specific examples of suitable organic acids include, but are not limited to, acetic acid, trifluoroacetic acid, formic acid, propionic acid, succinic acid, glycolic acid, gluconic acid, digluconic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, glucuronic acid, maleic acid, fumaric acid, pyruvic acid, aspartic acid, glutamic acid, benzoic acid, anthranilic acid, stearic acid, salicylic acid, p-hydroxybenzoic acid, phenylacetic acid, mandelic acid, pamoic acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, Pantothenic acid, toluenesulfonic acid, 2-hydroxyethanesulfonate, sufanilate, cyclohexylaminosulfonic acid, alginic acid, β-hydroxybutyric acid, galactonic acid, galacturonic acid, adipic acid, alginic acid, butyric acid, camphoric acid, camphorsulfonic acid, cyclopentanepropionic acid, dodecylsulfuric acid, saccharoheptanoic acid, glycerophosphate, heptanoic acid, hexanoic acid, nicotinic acid, 2-naphthalenesulfonic acid, oxalic acid, palmitic acid, pectic acid, 3-phenylpropionic acid, picric acid, pivalic acid, thiocyanic acid, and undecanoic acid.

[0142] In addition, when the compounds of the present disclosure carry an acidic moiety, suitable pharmaceutically acceptable salts thereof may include alkali metal salts, such as sodium or potassium salts; alkaline earth metal salts, such as calcium or magnesium salts; and salts formed with suitable organic ligands, such as quaternary ammonium salts. In another embodiment, base salts are formed from bases that form non-toxic salts, including aluminum salts, arginine salts, benzathine salts, choline salts, diethylamine salts, diethanolamine salts, glycine salts, lysine salts, meglumine salts, ethanolamine salts, tromethamine salts, and zinc salts.

[0143] Organic salts can be prepared from secondary, tertiary, or quaternary amines, such as tromethamine, diethylamine, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methylglucamine), and procaine. Basic nitrogen-containing groups can be quaternized with reagents such as lower alkyl (C1-Cs) halides (e.g., methyl, ethyl, propyl, and butyl chlorides, bromides, and iodides); dialkyl sulfates (e.g., dimethyl sulfate, diethyl sulfate, dibutyl sulfate, and diamyl sulfate); long-chain halides (e.g., decyl, lauryl, myristyl, and stearyl chlorides, bromides, and iodides); aralkyl halides (e.g., benzyl and phenethyl bromides), and the like.

[0144] In some cases, hemisalts of acids and bases may also be formed, such as hemisulphate and hemicalcium salts, or sequifumarate.

[0145] For a review of suitable salts, see "Handbook of Pharmaceutical Salts: Properties, Selection, and Use", Stahl and Wermuth (Wiley-VCH, 2002). Methods for preparing pharmaceutically acceptable salts of the disclosed compounds are known to those skilled in the art.

[0146] Other acids and bases, while not in themselves pharmaceutically acceptable, may be employed in the preparation of salts useful as intermediates in obtaining the compounds described herein and their pharmaceutically acceptable acid or base addition salts.

[0147] It should be understood that the compounds disclosed herein may exist as mixtures / combinations of different pharmaceutically acceptable salts. Mixtures / combinations of free forms of the compounds and pharmaceutically acceptable salts are also contemplated.

[0148] Compounds of the present disclosure can exist in a solid state continuum ranging from completely amorphous to completely crystalline. The term "amorphous" refers to a state in which a material lacks long-range order at the molecular level and can exhibit the physical properties of a solid or liquid depending on the temperature. Typically, such materials do not produce a unique X-ray diffraction pattern and, although exhibiting the properties of a solid, are more formally described as liquids. Upon heating, a change occurs from an apparent solid to a material having liquid properties, the change being characterized by a change in state, typically a secondary change ("glass transition"). The term "crystalline" refers to a solid phase in which a material has a regular, ordered internal structure at the molecular level and produces a unique X-ray diffraction pattern with defined peaks. Such a material will also exhibit the properties of a liquid when fully heated, but the change from solid to liquid is characterized by a phase transition, typically a primary phase transition ("melting point").

[0149] When subjected to suitable conditions, the compounds of the present disclosure can also exist in a mesomorphic state (mesophase or liquid crystal). The mesomorphic state is an intermediate between a true crystalline state and a true liquid state (melt or solution). The mesomorphism caused by temperature changes is described as "thermotropic", and the mesomorphism caused by adding a second component (such as water or another solvent) is described as "lyotropic". Compounds with the potential to form a lyotropic mesophase are described as "amphiphilic" and are composed of molecules with ionic (such as -COO'Na*, -COOK* or -SO3s'Na*) or nonionic (such as -N"N*(CHs)3) polar head groups. For more information, see Crystals and the Polarizing Microscope, NH Hartshorne and A. Stuart, 4th edition (Edward Arnold, 1970).

[0150] The present disclosure also relates to prodrugs of the disclosed compounds. Therefore, when certain derivatives of the disclosed compounds that may have little or no pharmacological activity are administered to the body or body, they can be converted into compounds of Formula I with the desired activity, for example, by hydrolytic cleavage. Such derivatives are referred to as "prodrugs." Further information on the use of prodrugs can be found in Pro-drugs as Novel Delivery Systems, Volume 14, ACS Symposium Series (T. Higuchi and W. Stella) and Bioreversible Carriers in Drug Design, Pergamon Press, 1987 (E.B. Roche, ed., American Pharmaceutical Association).

[0151] Prodrugs according to the present disclosure can be prepared, for example, by replacing appropriate functional groups present in the compounds of the present disclosure with certain moieties known to those skilled in the art as "promoieties," as described, for example, in Design of Prodrugs, H. Bundgaard (Elsevier, 1985) or in Prodrugs: Challenges and Reward, 2007 edition, edited by Valentino Stella, Ronald Borchardt, Michael Hageman, Reza Oliyai, Hans Maag, Jefferson Tilley, pp. 134-175 (Springer, 2007).

[0152] In addition, some compounds of the present disclosure itself can serve as prodrugs of other compounds of the present disclosure. The present disclosure also encompasses compounds of the present disclosure containing blocking groups. It will also be understood by those skilled in the art that compounds of the present disclosure can also be prepared with certain blocking groups, which can be used for purification or storage and can be removed before being administered to a patient. The protection and deprotection of functional groups are described in "Protective Groups in Organic Chemistry", edited by JWF McOmie, Plenum Press (1973) and "Protective Groups in Organic Synthesis", 3rd edition, TW Greene and PGM Wuts, Wiley-Interscience (1999).

[0153] Also included within the scope of the present disclosure are metabolites of the disclosed compounds, ie, compounds formed in vivo following administration of the drug.

[0154] pharmaceutical preparations

[0155] Also provided herein are pharmaceutical formulations comprising an effective amount of a compound of the present disclosure and one or more pharmaceutically acceptable excipients. As used herein, the term "formulation" is used interchangeably with "composition."

[0156] "Effective amount" includes "therapeutically effective amount" and "prophylactically effective amount". The term "therapeutically effective amount" refers to an amount that effectively treats and / or improves a disease or condition in a subject. The term "prophylactically effective amount" refers to an amount that effectively prevents a disease or condition and / or significantly reduces the chance of a disease or condition in a subject. As used herein, the terms "patient" and "subject" are used interchangeably and mean animals, such as dogs, cats, cattle, horses and sheep (i.e., non-human animals) and humans. A specific patient or subject is a mammal (e.g., a human). The terms "patient" and "subject" include males and females.

[0157] As used herein, the term "excipient" means any pharmaceutically acceptable additive, carrier, diluent, adjuvant or other ingredient other than the active pharmaceutical ingredient (API), appropriately selected according to the intended form of administration and in accordance with conventional pharmaceutical practice.

[0158] The compounds of the present disclosure may be administered alone or as part of a pharmaceutically acceptable composition or formulation. In addition, the compounds may be administered all at once, for example, by bolus injection, multiple times, for example, by a series of tablets, or delivered substantially evenly over a period of time, for example, using transdermal delivery. It should also be noted that the dosage of the compound may vary over time.

[0159] If desired, the compounds disclosed herein and other pharmaceutically active compounds can be administered to a subject or patient by any suitable route, such as orally, topically, rectally, parenterally (e.g., subcutaneous injection, intravenous, intramuscular, intrasternal, and intrathecal injection or infusion techniques), or as a buccal, inhalation, or nasal spray. Administration can provide a systemic effect (e.g., enteral or parenteral). All methods available to those skilled in the art for administering pharmaceutically active agents are contemplated. In some cases, the disclosed formulations can be administered orally or topically.

[0160] Suitable oral compositions or formulations according to the present disclosure include, but are not limited to, tablets, lozenges, troches, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, syrups or elixirs. Compositions or formulations suitable for oral use can be prepared according to any method known in the art for manufacturing pharmaceutical compositions.

[0161] Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs. In addition to the active compound, the liquid dosage form may also contain inert diluents commonly used in the art, such as water or other solvents, solubilizers, and emulsifiers, such as ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oil (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitan, and mixtures thereof. In addition to the inert diluent, oral compositions may also include adjuvants, such as wetting agents, emulsifiers and suspending agents, sweeteners, flavorings, and aromatics.

[0162] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is mixed with at least one inert, pharmaceutically acceptable excipient or carrier (such as sodium citrate or dicalcium phosphate) and / or the following substances: a) fillers or extenders (such as starch, lactose, sucrose, glucose, mannitol, and silicic acid; b) binders such as carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and gum arabic; c) humectants such as glycerol; d) disintegrants such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; e) solution retarder such as paraffin; f) absorption accelerators such as quaternary ammonium compounds; g) wetting agents such as cetyl alcohol and glyceryl monostearate; h) absorbents such as kaolin and bentonite; and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also include a buffer. The pharmaceutical composition may provide immediate, regulated or extended release of the compound, either alone or in combination. Examples of pharmaceutical compositions include, but are not limited to, immediate release, regulated release and extended release formulations.

[0163] Solid compositions of similar types can also be used as fillers in soft-filled and hard-filled gelatin capsules using excipients such as lactose or milk sugar and high molecular weight polyethylene glycols. Solid dosage forms of tablets, dragees, capsules, pills and granules can be prepared with coatings and shells such as enteric coatings and other coatings well known in the pharmaceutical formulation field. They can optionally contain opacifiers and can also be compositions that release the active ingredient only or preferentially in a certain part of the intestinal tract, optionally in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. Solid compositions of similar types can also be used as fillers in soft-filled and hard-filled gelatin capsules using excipients such as lactose or milk sugar and high molecular weight polyethylene glycols.

[0164] The active compound can also be in a microencapsulated form with one or more excipients as described above. Solid dosage forms such as tablets, lozenges, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, controlled-release coatings, and other coatings well known in the pharmaceutical formulation field. In such solid dosage forms, the active compound can be mixed with at least one inert diluent such as sucrose, lactose, or starch. As is conventional practice, such dosage forms can also contain other substances other than inert diluents, such as tableting lubricants and other tableting aids, such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets, and pills, the dosage form can also contain a buffer. They can optionally contain opacifiers and can also be compositions that release the active ingredient only or preferentially in a certain part of the intestinal tract, optionally in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes.

[0165] The pharmaceutical compositions and formulations described herein can also be administered topically or transdermally, particularly when the therapeutic target includes areas or organs readily accessible by topical application, including diseases of the eye, skin, or lower intestinal tract. Suitable topical formulations are readily prepared for each of these areas or organs. Topical application to the lower intestinal tract can be achieved, for example, with a rectal suppository formulation or with a suitable enema formulation. Dosage forms for topical or transdermal administration of the compounds described herein include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, suppositories, or patches.

[0166] For topical application, the pharmaceutical composition can be formulated into an active component suspended or dissolved in one or more carriers, and any desired preservatives or buffers that may be needed, suitable ointments, creams, lotions or gels. Carriers for topical application of the compound of the present invention include, but are not limited to, mineral oil, liquid paraffin, white vaseline, polyethylene glycol, polyoxyethylene, polyoxypropylene compounds, emulsifying wax and water. Alternatively, the pharmaceutical composition can be formulated into a suitable lotion or cream containing an active component suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl ester wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

[0167] Ophthalmic preparations, ear drops and eye drops are also encompassed within the scope of the present disclosure. In addition, the present disclosure contemplates the use of transdermal patches, which have the added advantage of providing controlled delivery of compounds to the body. Such dosage forms can be made by dissolving or dispersing the compound in a suitable medium. Absorption enhancers can also be used to increase the flow of the compound through the skin. Rate control can be achieved by providing a rate-controlling membrane or by dispersing the compound in a polymer matrix or gel.

[0168] Injectable preparations, such as sterile injectable aqueous or oily suspensions can be prepared using suitable dispersants or wetting agents and suspending agents according to known techniques. Sterile injectable preparations can also be sterile injectable solutions, suspensions or emulsions in nontoxic parenteral acceptable diluents or solvents, for example as solutions in 1,3-butanediol. Acceptable vehicles and solvents that can be used are water, Ringer's solution, USP and isotonic sodium chloride solution. In addition, sterile fixed oils are generally used as solvents or suspending media. For this purpose, any mild fixed oil can be used, including synthetic monoglycerides or diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of injections.

[0169] The injectable formulations can be sterilized, for example, by filtration through a bacteria-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

[0170] In order to prolong the effect of the compounds described herein, it is often desirable to slow down the absorption of the compound from subcutaneous or intramuscular injection. This can be achieved by using a liquid suspension of crystalline or amorphous materials with poor water solubility. The absorption rate of the compound then depends on its dissolution rate, which in turn may depend on the crystal size and crystalline form. Alternatively, delayed absorption of the compound form administered parenterally can be achieved by dissolving or suspending the compound in an oil vehicle. Depot injection preparations are prepared by forming a microcapsule matrix of the compound in a biodegradable polymer such as polylactic acid-polyglycolide. Depending on the ratio of the compound to the polymer and the properties of the specific polymer used, the rate of compound release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injection preparations can also be prepared by entrapping the compound in liposomes or microemulsions compatible with body tissues.

[0171] Compositions for rectal or vaginal administration are particularly suppositories, which can be prepared by mixing a compound described herein with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol or a suppository wax which are solid at ambient temperature but liquid at body temperature and therefore melt in the rectum or vaginal cavity and release the active compound.

[0172] Sterile injectable forms of the compositions described herein may be aqueous or oily suspensions. These suspensions can be formulated using suitable dispersants or wetting agents and suspending agents according to techniques known in the art. Sterile injectable preparations may also be sterile injectable solutions or suspensions in a nontoxic, parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Acceptable vehicles and solvents that may be used include water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile, fixed oils are commonly used as solvents or suspending media. For this purpose, any bland, fixed oil may be used, including synthetic mono- or diglycerides. Fatty acids, such as oleic acid and its glyceride derivatives, may be used to prepare injectables, as may natural pharmaceutically acceptable oils, such as olive oil or castor oil, particularly their polyoxyethylated forms. These oil solutions or suspensions may also contain long-chain alcohol diluents or dispersants, such as carboxymethylcellulose or similar dispersants, which are commonly used to formulate pharmaceutically acceptable dosage forms, including emulsions and suspensions. Other commonly used surfactants, such as Tweens, Spans and other emulsifying agents or bioavailability enhancers which are commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms may also be used for the purposes of formulation.

[0173] The pharmaceutical compositions can also be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well known in the art of pharmaceutical formulation and can be prepared as solutions in saline using benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and / or other conventional solubilizing or dispersing agents.

[0174] The compound for the method of the present disclosure can be formulated as a unit dosage form. The term "unit dosage form" refers to a physically discrete unit suitable as a unit dose for a subject being treated, each unit containing a predetermined amount of active substance calculated to produce a desired therapeutic effect, optionally together with a suitable pharmaceutical carrier. The unit dosage form can be a single daily dose or one of multiple daily doses (e.g., about 1 to 4 times or more per day). When multiple daily doses are used, the unit dosage form can be the same or different for each dose.

[0175] For oral administration, compositions can be provided in the form of tablets containing 0.01 milligram, 0.05 milligram, 0.1 milligram, 0.5 milligram, 1.0 milligram, 2.5 milligram, 5.0 milligram, 10.0 milligram, 15.0 milligram, 25.0 milligram, 50.0 milligram, 75.0 milligram, 100 milligram, 125 milligram, 150 milligram, 175 milligram, 200 milligram, 250 milligram and 500 milligram active ingredient, for adjusting the dosage of the patient according to symptoms. Medicine generally contains about 0.01 mg to about 500 mg of active ingredient, or in another embodiment, contains about 1 mg to about 100 mg of active ingredient. Intravenously, during constant rate infusion, dosage can be in the range of about 0.1 mg / kg / minute to about 10 mg / kg / minute.

[0176] Treatment

[0177] The compounds disclosed herein can act on the M4 receptor. The muscarinic acetylcholine receptor M4 (also known as muscarinic 4 or CHRM4) is a protein encoded by the CHRM4 gene in humans. The M4 receptor is primarily expressed in the brain. Key brain regions where M4 receptor expression occurs are the striatum, cortex, and hippocampus, with the highest expression (approximately 46%) occurring in the striatum, where M4 is the predominant muscarinic subtype. M4 is also sporadically expressed in the periphery (e.g., testes, skin, and colon).

[0178] M4 receptor and G q / iMuscarinic acetylcholine receptors are coupled to muscarinic acetylcholine receptors and function as inhibitory autoreceptors in the striatum and midbrain (Zhang et al.; "Multiple Muscarinic Acetylcholine Receptor Subtypes Modulate Striatal Dopamine Release, as Studied with M1–M5 Muscarinic Receptor Knock-Out Mice"; Journal of Neuroscience 2002 Aug 1, 22(15): 6347-6352; Tzavara et al.; "M4 muscarinic receptors regulate the dynamics of cholinergic and dopaminergic neurotransmission: relevance to the pathophysiology and treatment of related central nervous system pathologies"; FASEB Journal, 2004; 18: 1410-1412), and function as postsynaptic modulatory receptors in the striatum, neocortex, and hippocampus (Levey et al.; Identification and localization of muscarinic acetylcholine receptor proteins in brain with subtype-specific antibodies; Journal of Neuroscience October 1, 1991, 11 (10) 3218-3226; Gil et al.; "Muscarinic receptor subtypes in human iris-ciliary body measured by immunoprecipitation." Investigative ophthalmology & visual science 38.7 (1997): 1434-1442).M4 receptors have also been found presynaptically on glutamatergic synapses from the cortex to the striatum (Pancani, T. et al., “Allosteric activation of M4 improve behavioral and physiological alterations in early symptomatic YAC128 mice”, Proceedings of the National Academy of the Sciences of the United States of America, 2015 Nov 10;112(45):14078-83) and on hippocampal glutamate neurons where presynaptic M4 regulates glutamate release. The highest expression of M4 receptors is found in the striatum, where they also have a modulatory effect on dopaminergic neurotransmission and are co-expressed with D1 dopamine receptors in a subpopulation of striatal medium spiny neurons that contain GABA as the main neurotransmitter (Bernard et al. "Phenotypical characterization of the rat striatal neurons expressing muscarinic receptor genes"; Journal of Neuroscience 1992 Sep 1, 12(9) 3591-3600; DiChiara et al. "Modulatory functions of neurotransmitters in the striatum: ACh / dopamine / NMDA interactions"; Trends in Neurosciences; Vol. 17, No. 6, 1994, pp. 228-233; Ince et al. "Differential expression of D1 and D2 dopamine and m4 muscarinic acetylcholine receptor proteins in identified striatonigral neurons" neurons"; Synapse, (1997) 27:357-366).

[0179] It has been hypothesized that administration of selective M4 agonists would provide antipsychotic activity for the treatment of schizophrenia (Felder et al. "Elucidating the Role of Muscarinic Receptors in Psychosis", Life Sci. 68:2605-2613, 2001). This view is further supported by studies demonstrating that M4 receptors regulate the kinetics of dopaminergic and cholinergic neurotransmission and that dopamine hyperactivity states lead to loss of M4 function (Tzavara et al., 2004).

[0180] The compounds of the present disclosure may also be used to treat / alleviate neuropsychiatric symptoms (i.e., behavioral symptoms) associated with Alzheimer's disease and schizophrenia (Foster et al., "Activation of M1 and M4 muscarinicreceptors as potential treatments for Alzheimer's disease and schizophrenia", Neuropsychiatric Disease and Treatment; Vol. 2014, 10, pp. 183-191). These behavioral symptoms include, but are not limited to, agitation, vocal outbursts, compulsivity, anxiety, irritability, aggression, disorientation, hallucinations, delusions, hallucinations, suspicion, apathy, depression, disinhibition, abnormal movements and compulsive behavior, and sleep disorders (Dillon, Carol et al., "Behavioral symptoms related to cognitive impairment", Neuropsychiatric Disease and Treatment; 2013: 91443-1455). By treating / alleviating the above-mentioned behavioral symptoms, it is believed that the compounds of the present disclosure may also enhance cognition.

[0181] In view of the foregoing, the compounds of the present disclosure may be used to treat schizophrenia and Alzheimer's disease. The compounds of the present disclosure may also be used to treat Parkinson's disease, Huntington's disease, addiction, substance abuse disorders, depression, and epilepsy. The compounds of the present disclosure may also be used to treat Alzheimer's disease psychosis.

[0182] It is believed that the M4 selective activators of the present disclosure may also have a wide range of other therapeutic applications for treating central nervous system conditions or diseases, including neurological disorders, neurodegenerative disorders and / or psychiatric disorders. Neurological disorders, neurodegenerative disorders and / or psychiatric disorders include, but are not limited to, (1) mood [affective] disorders; (2) neurological, stress-related and somatoform disorders, including anxiety disorders; (3) disorders comprising symptoms of cognitive deficits in mammals (including humans); (4) disorders comprising attention deficits, executive function deficits (working memory deficits), impulse control dysfunction, extrapyramidal symptoms, disorders based on dysfunction of the basal ganglia, hippocampus and prefrontal cortex; (5) behavioral and emotional disorders that typically occur in childhood and adolescence; (6) psychological developmental disorders; (7) disorders primarily affecting the central nervous system. (8) extrapyramidal and movement disorders; (9) behavioral syndromes associated with physiological disturbances and physical factors; (10) personality and behavioral disorders in adults; (11) schizophrenia and other psychotic disorders; (12) mental and behavioral disorders due to the use of psychoactive substances; (13) sexual dysfunction including hypersexuality; (14) mental retardation; (15) factitious disorders, such as acute hallucinatory mania; (16) paroxysmal and epileptic disorders; (17) narcolepsy; (18) dementia, and (19) amyotrophic lateral sclerosis.

[0183] Examples of mood (affective) disorders that can be treated according to the present disclosure include, but are not limited to, bipolar disorder, hypomania (manic and mixed forms), bipolar II disorder; depressive disorders, such as a single depressive episode or recurrent major depressive disorder, chronic depression, psychotic depression, minor depressive disorder, postpartum onset depression, depressive disorder with psychotic symptoms; persistent mood [affective] disorders, such as cyclothymic temperament, dysthymia, euthymia; premenstrual syndrome (PMS) and premenstrual dysphoric disorder.

[0184] Examples of neurological, stress-related, and somatoform disorders that can be treated according to the present disclosure include, but are not limited to, anxiety disorders, social anxiety disorder, generalized anxiety disorder, panic disorder with or without agoraphobia, specific phobias, social phobia, chronic anxiety disorder; obsessive-compulsive disorder; reactions to severe stress and adjustment disorders such as post-traumatic stress disorder (PTSD), acute stress disorder, other neurological disorders such as depersonalization-derealization syndrome.

[0185] As used herein, the phrases "cognitive deficit" and "disorder comprising symptoms of cognitive deficit" refer to subnormal or suboptimal functioning in one or more aspects of cognition, such as memory, intelligence, learning and logical abilities, or attention and executive function (working memory), in a particular individual compared to other individuals in the same general age group.

[0186] Examples of "disorders comprising symptoms of cognitive deficits" that can be treated according to the present disclosure include, but are not limited to: cognitive deficits primarily but not exclusively associated with amnesia, psychosis (schizophrenia), Parkinson's disease, Alzheimer's disease, multi-infarct dementia, senile dementia, Lewy body dementia, stroke, frontotemporal dementia, progressive supranuclear palsy, Huntington's disease, HIV disease (HIV-associated dementia), brain trauma, and substance abuse; mild cognitive impairment ADHD, Asperger's syndrome, and age-related memory impairment; cognitive decline or delirium following surgery or associated with intensive care treatment.

[0187] Examples of disorders that are typically first diagnosed in infancy, childhood, and adolescence and that can be treated according to the present disclosure include, but are not limited to, hyperactivity disorders, including disturbances of activity and attention, attention-deficit / hyperactivity disorder (ADHD), hyperactive conduct disorder; attention deficit disorder (ADD); conduct disorders, including, but not limited to, depressive conduct disorder; tic disorders, including transient tic disorder, chronic motor or vocal tic disorder, combined vocal and polymotor tic disorder (Gilles de la Tourette's syndrome), substance-induced tic disorder; autism; Batten disease, excessive masturbation, nail biting, nose picking, and thumb sucking.

[0188] Examples of psychological developmental disorders that can be treated according to the present disclosure include, but are not limited to, pervasive developmental disorders, including but not limited to Asperger syndrome and Rett syndrome, autism, childhood autism and hyperactivity disorder associated with mental retardation and stereotyped movements, specific motor function developmental disorders, and specific academic skill developmental disorders.

[0189] Examples of systemic atrophies primarily affecting the central nervous system that may be treated according to the present disclosure include, but are not limited to, systemic atrophies due to multiple sclerosis primarily affecting the basal ganglia, including Huntington's disease, and amyotrophic lateral sclerosis.

[0190] Examples of extrapyramidal and movement disorders with basal ganglia dysfunction and / or degeneration that can be treated according to the present disclosure include, but are not limited to, Huntington's disease; Parkinson's disease; secondary parkinsonism, such as postencephalitic parkinsonism; other disorders including parkinsonism; Niemann-Pick disease, Lewy body disease; basal ganglia degenerative diseases; other extrapyramidal and movement disorders, including tremor, essential tremor and drug-induced tremor, myoclonus, chorea and drug-induced chorea, drug-induced tics and tics of organic origin, drug-induced acute dystonia, drug-induced tardive dyskinesia, muscle cramps and disorders associated with muscle spasticity or weakness (including tremor); mental impairment (including spasticity, Down syndrome and fragile X syndrome), levodopa-induced movement disorders; restless legs syndrome and stiff person syndrome.

[0191] Other examples of movement disorders with basal ganglia dysfunction and / or degeneration that can be treated according to the present disclosure include, but are not limited to, dystonia, including, but not limited to, focal dystonia, multifocal or segmental dystonia, torsion dystonia, hemispheric, generalized, and tardive dystonia (induced by psychopharmacological drugs). Focal dystonia includes cervical dystonia (torticollis), blepharospasm (eyelid spasm), limb dystonia (spasm of the limbs, such as writer's cramp), or mandibular dystonia and spasmodic dysphonia (vocal cord spasm); neuroleptic-induced movement disorders, including, but not limited to, neuroleptic malignant syndrome (NMS), neuroleptic-induced parkinsonism, neuroleptic-induced early onset or acute movement disorders, neuroleptic-induced acute dystonia, neuroleptic-induced acute akathisia, neuroleptic-induced tardive dyskinesia, and neuroleptic-induced tremor.

[0192] Examples of behavioral syndromes associated with physiological disturbances and physical factors according to the present disclosure include, but are not limited to, non-organic sleep disorders, including but not limited to non-organic hypersomnia, non-organic disturbances of sleep-wake schedules (circadian rhythm sleep disorders), insomnia, parasomnias, and sleep deprivation; mental and behavioral disorders associated with the puerperium, including postpartum and puerperal depression; eating disorders, including but not limited to anorexia nervosa, bulimia nervosa, eating disorders, excessive eating, obesity, compulsive eating disorders, and pagophagia.

[0193] Examples of adult personality and behavioral disorders that can be treated according to the present disclosure include, but are not limited to, personality disorders, including but not limited to emotional instability, borderline, obsessive-compulsive, compulsive, dependent, and passive-aggressive personality disorders; habit and impulse disorders (impulse control disorders), including intermittent explosive disorder, pathological gambling, pathological arson (pyromania), pathological stealing (kleptomania), trichotillomania; and Munchausen syndrome.

[0194] Examples of schizophrenia and other psychotic disorders that can be treated according to the present disclosure include, but are not limited to, different types of persistent or intermittent schizophrenia (e.g., delusional, juvenile, catatonic, undifferentiated, residual, and schizophreniform disorders); schizotypal disorders (such as borderline, latent, prepsychotic, prodromal, pseudoneurotic, pseudopsychotic schizophrenia, and schizotypal personality disorder); persistent delusional disorder; acute, transient, and persistent psychotic disorders; induced delusional disorder; different types of schizoaffective disorder (e.g., manic-depressive or mixed); puerperal psychosis and other and unspecified non-organic psychoses, such as social withdrawal in schizophrenia.

[0195] Examples of psychiatric and behavioral disorders due to the use of psychoactive substances that can be treated according to the present disclosure include, but are not limited to, psychiatric and behavioral disorders due to the use of alcohol, opioids, cannabinoids, sedatives or hypnotics, cocaine; psychiatric and behavioral disorders due to the use of other stimulants (including caffeine), psychiatric and behavioral disorders due to drug dependence and abuse (e.g., narcotic dependence, alcoholism, amphetamine and methamphetamine dependence, opioid dependence, cocaine addiction, nicotine dependence and drug withdrawal syndrome and relapse prevention), psychiatric and behavioral disorders due to the use of hallucinogens, tobacco (nicotine), volatile solvents, and psychiatric and behavioral disorders due to multiple drug use and use of other psychoactive substances, including the following subtypes: harmful use, dependence syndrome, withdrawal state, and withdrawal state with delirium.

[0196] Examples of dementias that may be treated according to the present disclosure include, but are not limited to, vascular dementia, dementia caused by Creutzfeld-Jacob disease, HIV, head trauma, Parkinson's disease, Huntington's disease, Pick's disease, and Alzheimer's type dementia.

[0197] Schizophrenia or psychosis for which the compounds of the present disclosure may be useful include one or more of the following conditions: schizophrenia (delusional, disorganized, catatonic, or undifferentiated), schizophreniform disorder, schizoaffective disorder, delusional disorder, brief psychotic disorder, shared psychotic disorder, psychotic disorder due to a general medical condition and substance-induced or drug-induced psychosis / psychotic disorder (phencyclidine, ketamine and other dissociative anesthetics, amphetamines and other psychostimulants and cocaine), psychosis associated with an affective disorder, brief reactive psychosis, schizoaffective psychosis, "psychosis," "Schizophrenia spectrum" disorders such as schizoid or schizotypal personality disorder, or disorders associated with psychosis (such as major depressive disorder, manic-depressive (bipolar) disorder, Alzheimer's disease, and post-traumatic stress syndrome), including positive and negative symptoms of schizophrenia and other psychotic disorders; cognitive impairment, including dementia (related to Alzheimer's disease, ischemic or multi-infarct dementia, trauma, vascular problems or stroke, HIV disease, Parkinson's disease, Huntington's disease, Pick's disease, Creutzfeldt-Jakob disease, perinatal hypoxia, other general medical conditions, or substance abuse); delirium, amnestic disorders, or age-related cognitive decline.

[0198] In addition to the central nervous system disorders mentioned above, the compounds of the present disclosure may also be used to treat other M4-mediated (or M4-related) disorders, such as, but not limited to, addiction (e.g., substance addiction, such as addiction to opioids, cocaine, or alcohol), pain (e.g., acute pain, inflammatory pain, and neuropathic pain), and sleep disorders (such as sleep disorders related to REM sleep regulation, e.g., sleep disorders related to REM sleep onset). Additional M4-mediated (or M4-related) disorders or conditions that can be treated with the compounds of the present disclosure include dry mouth, cognitive impairment (e.g., mild cognitive impairment), movement disorders, pulmonary hypertension, chronic obstructive pulmonary disease (COPD), asthma, urinary incontinence, glaucoma, trisomy 21 (Down syndrome), cerebral amyloid angiopathy, dementia (e.g., degenerative dementia), hereditary cerebral hemorrhage with amyloidosis of the Dutch type (HCHWA-D), Creutzfeldt-Jakob disease, prion disorders, amyotrophic lateral sclerosis, progressive supranuclear palsy, head trauma, stroke, pancreatitis, inclusion body myositis, other peripheral amyloidosis, diabetes, autism, and atherosclerosis. See, e.g., U.S. Patent No. 8,664,234.

[0199] Potential sleep disorders for which the compounds of the foregoing disclosure may be useful include: improving sleep quality; improving sleep quality; enhancing sleep maintenance; increasing the value calculated by dividing the time a subject sleeps by the time a subject attempts to sleep; reducing sleep latency or onset time (the time it takes to fall asleep); reducing difficulty falling asleep; improving sleep continuity; reducing the number of awakenings during sleep; reducing nighttime awakenings; reducing the time spent awake after the initial onset of sleep; increasing the total amount of sleep; reducing the fragmentation of sleep; altering the timing, frequency, or duration of REM sleep episodes; altering the timing, frequency, or duration of slow wave (i.e., stage 3 or 4) sleep episodes; increasing the amount and percentage of stage 2 sleep; promoting slow wave sleep; enhancing EEG-delta activity during sleep; improving Hypervigilance; reduction of daytime sleepiness; treatment or reduction of excessive daytime sleepiness; insomnia; hypersomnia; narcolepsy; sleep disruption; sleep apnea; insomnia; nocturnal myoclonus; REM sleep disruption; jet lag; shift worker sleep disorder; parasomnias; night terrors; insomnia associated with depression, affective / mood disorders, as well as sleepwalking and enuresis, and sleep disorders associated with aging; Alzheimer's sundowning; conditions related to circadian rhythmicity and mental and physical disorders associated with travel across time zones and shift work; conditions caused by drugs that cause decreased REM sleep as a side effect; syndromes characterized by non-restorative sleep and muscle pain or sleep apnea associated with breathing disturbances during sleep; and conditions caused by decreased sleep quality.

[0200] Pain disorders for which the compounds of the present disclosure may be useful include neuropathic pain (such as postherpetic neuralgia, nerve injury, "dynias," e.g., vulvodynia, phantom limb pain, radicular avulsions, painful diabetic neuropathy, painful traumatic mononeuropathy, painful polyneuropathy); central pain syndromes (which may result from virtually any injury at any level of the nervous system); postoperative pain syndromes (e.g., post-mastectomy syndrome, post-thoracotomy syndrome, stump pain); bone and joint pain (e.g., osteoarthritis), repetitive motion pain, dental pain, cancer pain, myofascial pain (muscle injury, fibromyalgia); perioperative pain (general surgery, gynecological pain), chronic pain, dysmenorrhea and pain associated with angina, and inflammatory pain of various origins (e.g., osteoarthritis, rheumatoid arthritis, rheumatic diseases, tenosynovitis and gout), headache, migraine and cluster headache, headache, primary hyperalgesia, secondary hyperalgesia, primary allodynia, secondary allodynia or other pain caused by central sensitization.

[0201] The compounds of the present disclosure are useful for reducing tolerance and / or dependence on opioids for the treatment of pain, and for treating withdrawal syndromes from, for example, alcohol, opioids, and cocaine.

[0202] In various instances, the M4-mediated (or M4-associated) disease or disorder can be selected from Alzheimer's disease, schizophrenia or psychosis, pain, addiction, sleep disorders, cognitive disorders (e.g., mild cognitive impairment), Parkinson's disease, Parkinson's disease-levodopa-induced dyskinesia, Huntington's disease, movement disorders, dry mouth, pulmonary hypertension, chronic obstructive pulmonary disease (COPD), asthma, urinary incontinence, glaucoma, trisomy 21 (Down syndrome), cerebral amyloid angiopathy, dementia, hereditary cerebral hemorrhage with amyloidosis Dutch type (HCHWA-D), Creutzfeldt-Jakob disease, prion disorders, amyotrophic lateral sclerosis, progressive supranuclear palsy, head trauma, stroke, pancreatitis, inclusion body myositis, other peripheral amyloidoses, diabetes, autism, and atherosclerosis. In some instances, the M4-mediated (or M4-associated) disease or disorder is selected from Alzheimer's disease, schizophrenia, pain, addiction, Parkinson's disease, Parkinson's disease-levodopa-induced dyskinesia, and sleep disorders.

[0203] Synthesis of Compounds of the Disclosure

[0204] The compounds of the present disclosure can be synthesized by any method known in the art. For example, the compounds of the present disclosure can be synthesized according to any one of Schemes 1-6.

[0205] Solution 1

[0206]

[0207] Scheme 1 relates to a synthetic sequence for preparing compounds of Formula I and Formula I'. According to Scheme 1, compound II can be coupled via Suzuki-Miyaura coupling reactions with heteroaryl, aryl, alkenyl, heterocycloalkyl, cycloalkyl, spiroheteroalkyl, spiroalkyl, alkyl and heteroalkyl boronic acids, borate esters or potassium trifluoroborate. The range of reaction types is not limited to Suzuki-Miyaura, but includes Stille, Negishi, Hiyama and decarboxylation couplings; Sonogashira couplings with alkynes / silylalkynes; and nucleophilic aromatic substitutions (S) of amines and alcohols. N Ar), where R 1 、R 2 、R 3 、R 4 and A substituents should be represented by the same moiety desired in the final product or its protected variants to be reacted with G 1 -R 3 Coupling reaction or S N Ar reacts to form compound III, while G 1 is boric acid / ester / trifluoroborate, stannane, magnesium, zinc, carboxylic acid, carboxylic acid ester; or, and G 1-R 3 is an alcohol / amine; or a terminal alkyne / silylalkyne, using standard choices of metal source, ligand, and base, in standard solvents / cosolvents, which include but are not limited to DMF, acetonitrile, 1,4-dioxane, THF, pyridine, toluene, ethanol, n-butanol, and tert-butanol. Examples of Pd / ligand / base combinations in coupling reactions include, but are not limited to, tetrakis(triphenylphosphine)palladium(0) plus sodium carbonate and tris(dibenzylideneacetone)dipalladium(0) plus dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl plus potassium carbonate. In S N In the Ar reaction, examples of bases include, but are not limited to, triethylamine, N,N-diisopropylethylamine, K2CO3, Cs2CO3, K3PO4, K3PO4.H2O, t BuOK and NaH. Removal of the protecting group P 1 Compound IV is obtained. Protective group P 1 In this case, it is a group known to those skilled in the art for amine protection. For example, P 1 It may be tert-butyloxycarbonyl (Boc), which can be cleaved via acidic conditions in a suitable solvent, including but not limited to treatment with a solution of trifluoroacetic acid (TFA) in dichloromethane (DCM). Alternatively, P 1 Can be one of many other protecting groups suitable for amines, including carboxybenzyl (Cbz) or benzoyl (Bz) groups, and can be cleaved under standard conditions known to those skilled in the art. Compound IV can be coupled with compound V (wherein m, n, and p are independently represented by integers selected from 1 or 2) to produce racemic compound VI using standard reductive amination procedures, such as, but not limited to, a combination of sodium cyanoborohydride and titanium (IV) ethoxide or sodium triacetoxyborohydride in a suitable solvent, followed by reaction with R in dichloromethane or other suitable solvent. 5 to produce a carbamate or carbamate bioisostere (where R 5 (which should be represented by the same moiety desired in the final product or a protected variant thereof) to produce a compound of Formula I as a racemic mixture. Chiral separation of the racemic mixture, for example, chiral chromatographic methods such as chiral HPLC or chiral supercritical fluid chromatography (SFC), can produce enantiomerically pure compounds of Formula I'.

[0208] Option 2

[0209]

[0210] Scheme 2 relates to an alternative synthetic route for preparing compounds of Formula I and Formula I'. Referring to Scheme 2a, compound IV (wherein R 1 、R 2 、R 3 、R4 and A substituent should be represented by the same moiety desired in the final product or a protected variant thereof) can be displaced in the presence of a base such as potassium carbonate or tripotassium phosphate in a suitable solvent (including but not limited to MeCN, DMSO, DMF or THF) to form a sulfonate of enantiomerically pure compound VII, wherein R 5 shall be represented by the same moiety as desired in the final product or its protected variant, wherein R 6 is an aryl, alkyl, fluoroaryl or fluoroalkyl substituent, such as 4-methylphenyl, methyl, nonafluorobutyl; and m, n and p are independently represented by integers selected from 1 or 2, and Y should be represented by the same moiety desired in the final product. 1 、R 2 、R 3 、R 4 and A should be represented by the same moiety desired in the final product or its protected variant) can be similarly replaced with the arylalkyl / fluoroalkyl sulfonate on chiral compound VIII to generate compound IX, wherein R 6 is an aryl, alkyl, fluoroaryl or fluoroalkyl substituent, such as 4-methylphenyl, methyl, nonafluorobutyl; and m, n and p independently represent an integer selected from 1 or 2, and Y should be represented by the same moiety desired in the final product. To remove the Boc group, the group can be cleaved via acidic conditions in a suitable solvent, including but not limited to trifluoroacetic acid in dichloromethane (DCM), followed by R 5 to obtain a carbamate or carbamate bioisostere (where R 5 (should be represented by the same moiety as desired in the final product or its protected variant) to produce a compound of Formula I' as a single enantiomer. Alternatively, as shown in Scheme 2c, compound IV (wherein R of Formula IV 1 、R 2 、R 3 、R 4 and the A substituent should be represented by the same moiety as desired in the final product or its protected variant) can be coupled with compound X (wherein R 5 (wherein R is the same moiety as that desired in the final product or its protected variant, m, n and p are independently represented by integers selected from 1 or 2, and Y is the same moiety as that desired in the final product) to produce compounds of Formula I as racemic mixtures using standard reductive amination procedures such as, but not limited to, a combination of sodium cyanoborohydride and titanium (IV) ethoxide or sodium triacetoxyborohydride in a suitable solvent. Chiral separation of racemates of Formula I can be accomplished by chiral HPLC or chiral SFC to provide compounds of Formula I' as single enantiomers. In Scheme 2d, compound XI (wherein R is the same moiety as that desired in the final product) is a chiral amination of compound XI as a racemic mixture. 1 、R 2 、R3 and R 4 shall be represented by the same moiety as desired in the final product or its protected variant, wherein G 2 is a sulfonate or a halogen) can be coupled with an enantiomerically pure compound XII (wherein A and R of formula XII are 5 shall be represented by the same moiety desired in the final product or a protected variant thereof, m, n and p are independently represented by an integer selected from 1 or 2, and Y shall be represented by the same moiety desired in the final product) to use standard coupling procedures such as, but not limited to, Suzuki-Miyaura coupling or Buchwald-Hartwig coupling reactions and S of amines. N Ar reacts to produce a compound of formula I'. In addition, referring to Scheme 2e, enantiomerically pure compound XIII can be coupled with heteroaryl, aryl, alkenyl, heterocycloalkyl, cycloalkyl, spiroheteroalkyl, spiroalkyl, alkyl and heteroalkyl boronic acid, borate or potassium trifluoroborate via Suzuki-Miyaura coupling reaction. The range of reaction types is not limited to Suzuki-Miyaura, but includes Stille, Negishi, Hiyama and decarboxylation coupling; Sonogashira coupling with alkynes / silylalkynes; and nucleophilic aromatic substitution (S) of amines and alcohols. N Ar) to produce Formula I' as a single enantiomer, wherein R 1 、R 2 、R 3 、R 4 、R 5 The and A substituents shall be represented by the same moiety as desired in the final product or its protected variants, m, n and p independently represent an integer selected from 1 or 2, and Y shall be represented by the same moiety as desired in the final product.

[0211] Option 3

[0212]

[0213] Scheme 3 relates to a synthetic sequence for preparing compounds of Formula Ia. According to Scheme 3, compound XIV can be coupled via Suzuki-Miyaura coupling reactions with heteroaryl, aryl, alkenyl, heterocycloalkyl, cycloalkyl, spiroheteroalkyl, spiroalkyl, alkyl and heteroalkyl boronic acids, borate esters or potassium trifluoroborate. The range of reaction types is not limited to Suzuki-Miyaura, but includes Stille, Negishi, Hiyama and decarboxylation couplings; Sonogashira couplings with alkynes / silylalkynes; and nucleophilic aromatic substitutions (S) of amines and alcohols. N Ar), and via coupling reaction or S NAr reaction, using standard selection of metal source, ligand and base, in standard solvents (such as but not limited to DMF, acetonitrile, 1,4-dioxane, THF, pyridine, toluene, ethanol, n-butanol, tert-butanol) produces compound XV, wherein R 1 、R 2 、R 3 and R 4 The substituents and X should be represented by the same moiety as desired in the final product or its protected variant. Examples of Pd / ligand / base combinations in coupling reactions include, but are not limited to, tetrakis(triphenylphosphine)palladium(0) plus potassium carbonate and tris(dibenzylideneacetone)dipalladium(0) plus dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl plus potassium carbonate. N In the Ar reaction, examples of bases include, but are not limited to, triethylamine, N,N-diisopropylethylamine, K2CO3, Cs2CO3, K3PO4, t-BuOK, and NaH. 1 Compound XVI is obtained. Protective group P 1 In this case, it is a group known to those skilled in the art for amine protection. For example, P 1 It may be tert-butyloxycarbonyl (Boc), which can be cleaved via acidic conditions in a suitable solvent, including but not limited to treatment with a solution of trifluoroacetic acid (TFA) in dichloromethane (DCM). Alternatively, P 1 Can be one of many other protecting groups suitable for amines, including carboxybenzyl (Cbz) or benzoyl (Bz) groups, and can be cleaved under standard conditions known to those skilled in the art. Compound XVI can be coupled with compound V (wherein m, n, and p are independently represented by integers selected from 1 or 2; and X and Y should be represented by the same moiety desired in the final product) to produce racemic compound XVII using standard reductive amination procedures, such as, but not limited to, a combination of sodium cyanoborohydride and titanium (IV) ethoxide or sodium triacetoxyborohydride in a suitable solvent, followed by deprotection of the Boc-protecting group and reaction with R 5 to produce a carbamate or carbamate bioisostere (where R 5 should be represented by the same moiety desired in the final product or a protected version thereof) to produce a compound of Formula Ia.

[0214] Option 4

[0215]

[0216] Scheme 4 relates to a synthetic sequence for preparing compounds of Formula Ib. Referring to Scheme 4, compound XVIII can be coupled via Suzuki-Miyaura coupling reactions with heteroaryl, aryl, alkenyl, heterocycloalkyl, cycloalkyl, spiroheteroalkyl, spiroalkyl, alkyl and heteroalkylboronic acids, borate esters or potassium trifluoroborate. The range of reaction types is not limited to Suzuki-Miyaura, but includes Stille, Negishi, Hiyama and decarboxylation couplings; Sonogashira couplings with alkynes / silylalkynes; and nucleophilic aromatic substitutions (S) of amines and alcohols. N Ar), and via coupling reaction or S N Ar reaction, using standard selection of metal source, ligand and base, in standard solvents (such as but not limited to DMF, acetonitrile, 1,4-dioxane, THF, pyridine, toluene, ethanol, n-butanol, tert-butanol) produces compound XIX, wherein R 1 、R 2 、R 3 and R 4 Substituents should be represented by the same moiety as desired in the final product or its protected variant. Examples of Pd / ligand / base combinations in coupling reactions include, but are not limited to, tetrakis(triphenylphosphine)palladium(0) plus sodium carbonate and tris(dibenzylideneacetone)dipalladium(0) plus dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl plus potassium carbonate. N In the Ar reaction, examples of bases include, but are not limited to, triethylamine, N,N-diisopropylethylamine, K2CO3, Cs2CO3, K3PO4, t BuOK and NaH. Removal of the protecting group P 1 Obtain compound XX. Protective group P 1 In this case, it is a group known to those skilled in the art for amine protection. For example, P 1 It may be tert-butyloxycarbonyl (Boc), which can be cleaved via acidic conditions in a suitable solvent, including but not limited to treatment with a solution of trifluoroacetic acid (TFA) in dichloromethane (DCM). Alternatively, P 1 The protecting group B may be one of many other protecting groups suitable for amines, including carboxybenzyl (Cbz) or benzoyl (Bz) groups, and may be cleaved under standard conditions known to those skilled in the art. Compound XX may be coupled with compound XXI to produce racemic compound XXII using standard reductive amination procedures, such as, but not limited to, a combination of sodium cyanoborohydride and titanium (IV) ethoxide or sodium triacetoxyborohydride in a suitable solvent, followed by deprotection of the Boc group and reaction with R in dichloromethane or other suitable solvent. 5 to produce a carbamate or carbamate bioisostere (where R 5shall be represented by the same moiety as desired in the final product or a protected version thereof) to produce a compound of Formula Ib.

[0217] Option 5

[0218]

[0219] Scheme 5 relates to an alternative synthetic route for preparing compounds of formula Ic. Referring to Scheme 5a, compound XXVII can be prepared via two synthetic methods. One synthetic method is to prepare enantiomerically pure compound XXIII, wherein R 6 Should be represented by an aryl, alkyl, fluoroaryl or fluoroalkyl substituent, for example 4-methylphenyl, methyl or nonafluorobutyl can be coupled with XXIV in the presence of a base such as potassium carbonate or tripotassium phosphate in a suitable solvent (including but not limited to MeCN, DMSO, DMF or THF). Alternatively, compound XXVII can be prepared by the bis-S reaction of enantiomerically pure amine XXV and dichloro XXVI. N 2. Reaction preparation. Removal of protective group P 1 Compound XXVIII is obtained. Protective group P 1 In this case, it is a group known to those skilled in the art for amine protection. For example, P 1 It may be a carboxybenzyl group (Cbz), which can be cleaved via H2 gas conditions in an appropriate solvent, including but not limited to treatment with a wet 10% Pd / C solution in methanol (MeOH). Alternatively, P 1 Can be one of many other protecting groups suitable for amines, including benzyl (Bn) or benzoyl (Bz) groups, and can be cleaved under standard conditions known to those skilled in the art. Compound XXVIII can be coupled with compound XI (wherein R 1 、R 2 、R 3 and R 4 shall be represented by the same moiety as desired in the final product or its protected variant, wherein G 2 is a sulfonate or halogen) to react with an amine using standard CN coupling procedures such as, but not limited to, a Buchwald-Hartwig coupling reaction or S-coupling of an amine as appropriate. N Ar reaction to produce the compound of formula XXIX. Subsequently, the tert-butyloxycarbonyl (Boc) group of compound XXIX can be cleaved by acidic conditions in a suitable solvent, including but not limited to treatment with a solution of trifluoroacetic acid (TFA) in dichloromethane (DCM) to provide the enantiomeric compound XXX, which is then treated with R in dichloromethane or other suitable solvents. 5 to produce a carbamate or carbamate bioisostere (where R 5(should be represented by the same moiety as desired in the final product or its protected variant) to produce a compound of Formula Ic. Alternatively, as shown in Scheme 5b, compound XXXI (wherein R 1 、R 2 、R 3 and R 4 should be represented by the same moiety as desired in the final product or its protected variant) can be represented by double S N 2 is coupled with XXVI to generate the cyclized compound XIX. 1 Obtain compound XX. Protective group P 1 In this case, it is a group known to those skilled in the art for amine protection. For example, P 1 It may be tert-butyloxycarbonyl (Boc), which can be cleaved via acidic conditions in a suitable solvent, including but not limited to treatment with a solution of trifluoroacetic acid (TFA) in dichloromethane (DCM). Alternatively, P 1 R may be one of many other protecting groups suitable for amines, including carboxybenzyl (Cbz) or benzoyl (Bz) groups, and may be cleaved under standard conditions known to those skilled in the art. Compound XX may displace the enantiomerically pure sulfonate of compound XXIII, wherein R 6 is an aryl, alkyl, fluoroaryl or fluoroalkyl substituent, for example, 4-methylphenyl, methyl, nonafluorobutyl; for example, 4-methylphenyl or methyl in the presence of a base such as potassium carbonate in a suitable solvent (including but not limited to DMSO, DMF or THF) to generate compound XXIX. Subsequently, the tert-butyloxycarbonyl (Boc) group of compound XXIX can be cleaved via acidic conditions in a suitable solvent, including but not limited to treatment with a solution of trifluoroacetic acid (TFA) in dichloromethane (DCM) to provide enantiomeric compound XXX, which is then further treated with R in dichloromethane or other suitable solvents. 5 is processed to produce a carbamate or carbamate bioisostere, wherein R 5 The same moiety as desired in the final product or its protected variant should be represented to produce the compound of Formula Ic.

[0220] Option 6

[0221]

[0222] Scheme 6 relates to a synthetic sequence for preparing compounds of Formula Id. Referring to Scheme 6a, compound XXX (wherein R 1 、R 2 、R 3 and R 4The substituents should be represented by the same moiety as desired in the final product or its protected variants) can then be treated with CO gas, a base such as DBU and a halide / sulfonate-Z or Z--sulfate-Z (wherein Z should be represented by the same moiety as desired in the final product or its protected variants) in DMF or other suitable solvents to generate compounds of Formula Id. An alternative synthetic route to prepare Formula Id is to use triphosgene and ZOH in a suitable solvent such as DCM and a base such as pyridine (wherein Z should be represented by the same moiety as desired in the final product or its protected variants) to react with compound XXX to synthesize the carbamate of Formula Id. Another alternative method for preparing Formula Id is to use CDI (instead of triphosgene) and ZOH (wherein Z should be represented by the same moiety as desired in the final product or its protected variants) to react with compound XXX to synthesize the carbamate of Formula Id. In Scheme 6b, the tert-butyloxycarbonyl (Boc) group of compound XXVII can be cleaved via acidic conditions in a suitable solvent, including but not limited to treatment with a solution of trifluoroacetic acid (TFA) in dichloromethane (DCM) to provide enantiomeric compound XXXI, followed by three reaction methods to obtain compound XXXII, wherein Z should be represented by the same moiety as desired in the final product or its protected variant. The three methods are: 1) treatment with CO2 gas, a base such as DBU, and Z-sulfate-Z or halide / sulfonate-Z in a suitable solvent such as DMF; 2) use of triphosgene and ZOH in a suitable solvent such as DCM and a base such as pyridine; 3) use of CDI and ZOH in a suitable solvent such as THF. Removal of the protecting group P 1 Compound XXXIII is obtained. Protective group P 1 In this case, it is a group known to those skilled in the art for amine protection. For example, P 1 It may be a carboxybenzyl group (Cbz), which can be cleaved via H2 gas conditions in an appropriate solvent, including but not limited to treatment with a wet 10% Pd / C solution in methanol (MeOH). Alternatively, P 1 Can be one of many other protecting groups suitable for amines, including benzyl (Bn) or benzoyl (Bz) groups, and can be cleaved under standard conditions known to those skilled in the art. Compound XXXIII can be coupled with compound XI (wherein R 1 、R 2 、R 3 and R 4 shall be represented by the same moiety as desired in the final product or its protected variant, wherein G 2 is a sulfonate or halogen) to react with the amine using standard CN coupling procedures such as, but not limited to, Buchwald-Hartwig coupling reactions and S-coupling of amines as appropriate. N Ar reacts to produce a compound of formula Id.

[0223] Implementations of the Disclosure

[0224] 1. A compound having a structure of formula (I):

[0225]

[0226] or an N-oxide thereof, or a pharmaceutically acceptable salt of said compound or its N-oxide, wherein:

[0227] A is a ring containing 1 or 2 nitrogen atoms and surrounded by 0, 1, 2 or 3 R A substituted 6-8 membered heterocycle;

[0228] Each R A Independently C 1-3 Alkyl, halogen, =O, OH, C 1-3 Hydroxyalkyl or C 1-3 alkyl halide;

[0229] Y is a bond, S, O, CH2, CHF, CF2 or C(OH)H;

[0230] m is 1 or 2;

[0231] n is 1 or 2;

[0232] p is 1 or 2;

[0233] R 1 H, halogen, CN, OH, NO2, -N(R 6 )(R 7 ), C 1-6 Alkyl, C 2-6 Heteroalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, -[O] 0-1 -C 3-6 Cycloalkyl, -[O] 0-1 -C 6-10 Aryl, -[O] 0-1 -4-8 membered heterocycle or -[O] 0-1 -5-10 membered heteroaryl, wherein the heterocycle and heteroaryl each contain 1, 2 or 3 ring heteroatoms selected from N, O and S, and when R 1 C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, -[O] 0-1 -C 3-6 Cycloalkyl, -[O] 0-1 -C 6-10 Aryl, -[O] 0-1-4-8 membered heterocyclic ring, -[O] 0-1 -5-10 membered heteroaryl, -NH-C 3-6 Cycloalkyl, -NH-C 6-10 Aryl, -NH-4-8 membered heterocyclic ring, -NH-5-10 membered heteroaryl, -N(C 1-6 alkyl)-C 3-6 Cycloalkyl, -N(C 1-6 alkyl)-C 6-10 Aryl, -N(C 1-6 alkyl)-4-8 membered heterocyclic ring or -N(C 1-6 alkyl)-5-10 membered heteroaryl, R 1 0, 1, 2 or 3 independently selected from halogen, CN, OH, ═O, SO2 and C 1-3 Substitution of alkyl groups;

[0234] R 2 H, halogen, CN, OH, –N(R 6 )(R 7 ), C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 2-6 Heteroalkyl, C 3-6 cycloalkyl or a 4-8 membered heterocycle containing 1, 2 or 3 ring heteroatoms selected from N, O and S;

[0235] R 3 Halogen, C 0-6 Alkylene -CN, OH, -N(R 6 )(R 7 ), C 1-6 Alkyl, C 2-6 Heteroalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, C 1-6 Alkylene-OC 1-6 Alkyl, C 1-6 Alkylene-NHC(O)C 1-6 Alkyl, C 1-6 Alkylene-C(O)NHC 1-6 Alkyl, C 0-6 Alkylene-Cyc, OC 0-6 Alkylene-Cyc, NH-Cyc, N(C 1-6 alkyl)-Cyc or C(O)Cyc;

[0236] Cyc is C 3-12 Cycloalkyl, C 3-6 Heterocycloalkyl, C5-12 Spiroalkyl, C 5-12 Heterospirocycloalkyl, C 6-10 aryl, 4-12 membered heterocyclic or 5-10 membered heteroaryl, said heterocyclic or heteroaryl containing 1, 2 or 3 ring heteroatoms independently selected from N, O and S, and Cyc is replaced by 0, 1, 2 or 3 R 3a Substituent substitution;

[0237] Each R 3a independently selected from halogen, CN, OH, ═O, ═N(C 1-6 alkyl), SO2, C 1-6 Alkyl, C 2-10 Olefins, C 1-6 Hydroxyalkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 1-6 Alkylene-OC 1-6 Alkyl, C 0-6 Alkylene-C(O)C 1-6 Alkyl, C 0-6 Alkylene-NH2, C 0-6 Alkylene-NH(C 1-6 Alkyl), C 0-6 Alkylene-N(C 1-6 Alkyl)2, -SC 1-6 Alkyl, C 0-6 Alkylene-SO2C 1-6 Alkyl, C 0-6 Alkylene-C(O)NH2, C 0-6 Alkylene-C(O)NH(C 1-6 Alkyl), C 0-6 Alkylene-C(O)N(C 1-6 Alkyl)2, C 0-6 Alkylene-NHC(O)C 1-6 Alkyl, C 0-6 Alkylene-COOH, C 0-6 Alkylene-CO2C 1-6 Alkyl, C 0-6 Alkylene-C 3-6 Cycloalkyl and C containing 1, 2 or 3 heteroatoms selected from N, O and S 0-6 Alkylene-3-6 membered heterocycle;

[0238] R 4 H, halogen, CN, OH, C 1-6 Alkyl, C 1-6 -haloalkyl or C 1-6 alkoxy;

[0239] R5 is -CO2-Z or its bioisostere;

[0240] Each R 6 and R 7 Independently H, C 1-6 Alkyl, C(O)-C 1-6 Alkyl, spiro or bicyclic C 8-14 cycloalkyl, an 8-14 membered heterocycle containing 1, 2 or 3 ring heteroatoms selected from N, O and S, and when R 6 or R 7 When not H, it is replaced by 0, 1, 2 or 3 independently selected from halogen, CN, ═O, SO 2 , OH, C 0-6 Alkylene-NH2, C 0-6 Alkylene-NH(C 1-6 Alkyl), C 0-6 Alkylene-N(C 1-6 Alkyl)2, C 0-6 Alkylene-SO2C 1-6 Alkyl, C 1-6 Alkyl and C 1-6 The alkoxy substituent is substituted, or

[0241] R 6 and R 7 together with the nitrogen to which they are attached, form a 4-10 membered heterocyclic ring containing 0-2 additional ring heteroatoms independently selected from N, O and S; and

[0242] Z is C 1-7 Alkyl, C 1-7 Halogenated alkyl, C 3-6 Cycloalkyl or C 2-6 Alkyne, and Z is optionally replaced by C 1-6 Alkoxy or C 3-6 cycloalkyl substitution;

[0243] The condition is that when R 1 and R 4 Each is H, m and p are each 1, and A is and

[0244] (a)R 2 is H, Y is CH2, n is 1, and R 3 CH3, OCH3, or

[0245] (b)R 2 is F, Y is CH2, n is 1, and R 3 for or

[0246] (c)R 2 is H, Y is a bond, n is 1, and R 3 for or

[0247] (d)R 2 is H, Y is a bond, n is 2, and R 3 for When R 5 Not CO2CH2CH3 or CO2CH(CH3)3.

[0248] 2. The compound or salt according to claim 1, wherein A is and X is N, CH, C(OH) or CF.

[0249] 3. The compound or salt according to embodiment 2, which has a structure of formula (Ia), or a pharmaceutically acceptable salt thereof:

[0250]

[0251] 4. The compound or salt of any one of Embodiments 1 to 3, wherein Y is CH2, CHF, CF2 or C(OH)H.

[0252] 5. The compound or salt according to embodiment 4, wherein Y is CH2.

[0253] 6. The compound or salt according to any one of Embodiments 1 to 5, wherein m is 1.

[0254] 7. The compound or salt according to any one of Embodiments 1 to 6, wherein n is 1.

[0255] 8. The compound or salt of any one of Embodiments 1 to 7, wherein p is 1.

[0256] 9. The compound or salt according to embodiment 1, which has the structure of formula (Ib):

[0257]

[0258] 10. The compound or salt according to embodiment 9, which has the structure of formula (Ic):

[0259]

[0260] 11. The compound or salt according to embodiment 9, which has the structure of formula (Id):

[0261]

[0262] 12. The compound or salt according to embodiment 1, which has a structure of Formula (Ie), or a pharmaceutically acceptable salt thereof:

[0263]

[0264] 13. A compound or salt according to any one of embodiments 1 to 11, wherein R 5 is a CO2Z bioisostere and is selected from

[0265] 14. A compound or salt according to any one of embodiments 1 to 12, wherein R 5 Selected from CO2C 1-7 alkyl,

[0266] 15. The compound or salt according to embodiment 14, wherein R 5 It is CO2CH2CH3.

[0267] 16. A compound or salt according to any one of embodiments 1 to 15, wherein R 1 H, halogen, CN, OH, -N(R 6 )(R 7 ), C 1-6 Alkyl or C 1-6 Alkoxy.

[0268] 17. The compound or salt according to embodiment 16, wherein R 1 is H or halogen.

[0269] 18. A compound or salt according to any one of embodiments 1 to 17, wherein R 2 H, halogen, CN, OH, -N(R 6 )(R 7 ), C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy or C 1-6 Halogenated alkoxy.

[0270] 19. The compound or salt according to embodiment 18, wherein R 2 H, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl or C 1-6 Halogenated alkoxy.

[0271] 20. The compound or salt according to embodiment 19, wherein R 2 is H or halogen.

[0272] 21. A compound or salt according to any one of embodiments 1 to 20, wherein each R 6 and R 7 Independently H, C 1-6 Alkyl or C(O)-C 1-6 alkyl.

[0273] 22. The compound or salt according to embodiment 21, wherein each R 6 and R 7 are independently H or C 1-6 alkyl.

[0274] 23. A compound or salt according to any one of embodiments 1 to 20, wherein at least one R 6 and R 7 Together with the nitrogen to which they are attached they form a 4-10 membered heterocyclic ring containing 0-2 additional ring heteroatoms independently selected from N and O.

[0275] 24. A compound or salt according to any one of embodiments 1 to 23, wherein R 4 is H or halogen.

[0276] 25. A compound or salt according to any one of embodiments 1 to 24, wherein R 1 、R 2 and R 4 At least one of them is a halogen.

[0277] 26. A compound or salt according to any one of embodiments 1 to 25, wherein R 1 、R 2 and R 4 At least one of them is F.

[0278] 27. A compound or salt according to any one of embodiments 1 to 26, wherein R 3 For -[O] 0-1 -C 3-6 Cycloalkyl, -[O] 0-1 -C 6-10 Aryl, -[O] 0-1 -4-8 membered heterocycle or -[O] 0-1 -5-10 membered heteroaryl, and R 3 0, 1, 2 or 3 R 3a replace.

[0279] 28. The compound or salt according to embodiment 27, wherein R 3 C3-6 cycloalkyl, 5-10 membered heteroaryl or 4-8 membered heterocycle, and R 3 0, 1, 2 or 3 R 3a replace.

[0280] 29. A compound or salt according to any one of embodiments 1 to 26, wherein R 3 for and 0, 1, 2 or 3 R 3a replace.

[0281] 30. The compound or salt according to embodiment 29, wherein R 3 for and 0, 1, 2 or 3 R 3a replace.

[0282] 31. A compound or salt according to any one of embodiments 1 to 30, wherein R 3 is unsubstituted.

[0283] 32. A compound or salt according to any one of embodiments 1 to 30, wherein R 3 By 1 or 2 R 3a replace.

[0284] 33. The compound or salt according to embodiment 32, wherein R 3 By 1 R 3a replace.

[0285] 34. A compound or salt according to any one of embodiments 1 to 30, 32 and 33, wherein at least one R 3a Halogen, CN, OH, =O, SO2, C 1-6 Alkyl, C 2-10 Olefins, C 1-6 Hydroxyalkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 1-6 Alkylene-OC 1-6 Alkyl, N(C 1-6 Alkyl)2, C 1-6 Alkylene-N(C 1-6 Alkyl)2, -SC 1-6 Alkyl, NHC(O)C 1-6 Alkyl, C 1-6 Alkylene-NHC(O)C 1-6an alkyl group or a C 0-6 Alkylene-3-6 membered heterocyclic ring.

[0286] 35. The compound or salt according to embodiment 34, wherein at least one R 3a For CH3, CH2CH3, CH(CH3)2, CH2CH(CH3)2, CF3, CH2CH2F, CH2CHF2, CH2OH, C(CH3)2OH, CH2OCH3, CH2CH2OCH3, CH2O CH2CH3, F, CN, =O, SO2, OH, OCH3, OCH2CH3, OCH(CH3)2, OCHF2, CH2OCH3, CH2OCF3, SCH3, N(CH3)2, NHCOCH3, CD3,

[0287] 36. The compound or salt according to embodiment 35, wherein at least one R 3a is CH3, CH2CH3, F, CN, OH, OCH3, CF3, CH2OH or OCHF2.

[0288] 37. A compound as described in Table A or a pharmaceutically acceptable salt thereof.

[0289] 38. The salt of embodiment 37, wherein the salt is selected from HCl, HCl.H2O, maleate, and maleate.H2O.

[0290] 39. A pharmaceutical formulation comprising a therapeutically effective amount of a compound or salt according to any one of Embodiments 1 to 38, and a pharmaceutically acceptable excipient.

[0291] 40. A method for treating an M4-mediated (or M4-related) disease or disorder in a patient, the method comprising administering to the patient a therapeutically effective amount of a compound or salt according to any one of Embodiments 1 to 38.

[0292] 41. The method of embodiment 40, wherein the M4-mediated (or M4-related) disease or disorder is selected from Alzheimer's disease, schizophrenia or psychosis, pain, addiction, sleep disorders, cognitive disorders (e.g., mild cognitive impairment), Parkinson's disease, Parkinson's disease-levodopa-induced dyskinesia, Huntington's disease, movement disorders, dry mouth, pulmonary hypertension, chronic obstructive pulmonary disease (COPD), asthma, urinary incontinence, glaucoma, trisomy 21 (Down syndrome), cerebral amyloid angiopathy, Alzheimer's disease psychosis, dementia-related psychosis, bipolar disorder, hereditary cerebral hemorrhage with amyloidosis Dutch type (HCHWA-D), Creutzfeldt-Jakob disease, prion disorders, amyotrophic lateral sclerosis, progressive supranuclear palsy, head trauma, stroke, pancreatitis, inclusion body myositis, other peripheral amyloidosis, diabetes, autism, and atherosclerosis.

[0293] 42. The method of embodiment 41, wherein the M4-mediated (or M4-related) disease or disorder is selected from Alzheimer's disease, schizophrenia, pain, addiction, Parkinson's disease, Parkinson's disease-levodopa-induced movement disorder, and sleep disorder.

[0294] Example

[0295] The following examples are provided for illustration and are not intended to limit the scope of the present disclosure.

[0296] As used throughout these examples, common organic abbreviations are defined as follows:

[0297]

[0298]

[0299] An inert atmosphere (nitrogen or argon) is generally required, particularly when oxygen or moisture-sensitive reagents such as dried Pd / C, intermediates, and / or inert conditions are employed. Commercial solvents and reagents are generally used without further purification. Where appropriate, anhydrous solvents are employed, typically from WuXi-EHS, meeting the following QC specifications for water: a) <50 ppm for N'N-dimethylformamide; b) <100 ppm for dichloromethane, toluene, and tetrahydrofuran; c) <200 ppm for methanol, ethanol, 1,4-dioxane, and diisopropylamine. The product is typically dried under vacuum prior to further reaction or bioassay. Mass spectral data are reported by liquid chromatography-mass spectrometry (LCMS) and high performance liquid chromatography (HPLC). 1H NMR spectra were recorded at 400 MHz on a Bruker instrument. The chemical shift (δ) of nuclear magnetic resonance (NMR) data is expressed as parts per million (ppm) relative to the residual peak from the deuterated solvent employed. In addition, chiral separations were performed to separate the enantiomers of certain compounds of the present invention by supercritical fluid chromatography (SFC). In some examples, the enantiomers separated were designated as peak 1 and peak 2 according to their elution order. Based on effectiveness, chirality can be realized as (R)-isomers or (S)-isomers. LCMS is typically performed after the reaction by detectable intermediates, and is performed to complete conversion before subsequent reagents are added.

[0300] For the synthetic reference procedures in other embodiments or methods, reaction conditions (reaction time and temperature) can vary. For some compounds, microwave-mediated reactions are carried out in a Biotage Initiator microwave reactor. Typically, the reactions are monitored by thin-layer chromatography or mass spectrometry, and aftertreatment is performed where appropriate. Purification can vary between experiments: in general, solvents and solvent ratios for eluent / gradient are selected to provide suitable Rf or retention time. All starting materials in these preparations and embodiments are commercially available or can be prepared by methods known in the art or as described herein.

[0301] LCMC method 1: Instrument: SHIMADZU LC20-MS2010; Mobile phase: 1.5 mL / 4 L TFA in water (solvent A) and 0.75 mL / 4 L TFA in ACN (solvent B), using an elution gradient of 5%-95% (solvent B) in 0.7 minutes and holding at 95% for 0.4 minutes at a flow rate of 1.5 mL / min; Column: MERCK, RP-18e 25-2 mm; Wavelength: UV 220 nm and 254 nm; Column temperature: 50°C; MS ionization: ESI.

[0302] LCMC method 2: Instrument: SHIMADZU LC20-MS2020; Mobile phase: 0.8 mL / 4 L NH3·H2O in water (solvent A) and ACN (solvent B), using an elution gradient of 10%-80% (solvent B) in 6 minutes and holding at 80% for 0.5 minutes at a flow rate of 0.8 mL / min; Column: Titank C18; 5 μm; 2.1×50 mm; Wavelength: UV 220 nm and 254 nm; Column temperature: 50°C; MS ionization: ESI.

[0303] LCMC method 3: Instrument: SHIMADZU LCMS-2020; Mobile phase: 5% ACN (0.01875% TFA) in water (0.0375% TFA) to 95% ACN in water over 0.60 min, with a flow rate set at 2.0 mL / min; then held at 95% ACN for 0.18 min, with a flow rate set at 2.0 mL / min; returned to 5% ACN in water and held for 0.02 min, with a flow rate set at 2.0 mL / min; Column: EVO C18 2.1×30mm, 5µm. Column temperature: 50°C.

[0304] HPLC method 1: instrument: SHIMADZU LC20-MS2020; mobile phase: 0.2 mL / 1 L NH3·H2O in water (solvent A) and ACN (solvent B), using an elution gradient of 10%-80% (solvent B) within 6 minutes and holding at 80% for 2 minutes at a flow rate of 0.8 mL / min; column: TitankC18; 5 μm; 2.1×50 mm; wavelength: UV220 nm, 215 nm and 254 nm; column temperature: 50°C.

[0305] HPLC Method 2: Instrument: SHIMADZU LC-20AD; Mobile Phase: 10% ACN (0.018% TFA) in water (0.037% TFA) to 80% ACN in water over 3.00 minutes, flow rate set at 1.5 mL / min; then hold at 80% ACN for 0.70 minutes, flow rate set at 1.5 mL / min; return to 10% ACN in water and hold for 0.30 minutes, flow rate set at 2.0 mL / min; Column: Kinetex C18 LC column, 4.6 × 50 mm, 5 μm. Wavelengths: UV 220 nm and 254 nm. Column Temperature: 50°C.

[0306] SFC method 1: instrument: CAS-SH-ANA-SFC-G (Agilent 1260 with DAD detector); column: ChiralPak AD-3150×4.6 mm ID, 3 um; mobile phase: A: CO2, B: methanol (0.05% DEA) isocratic: 40% B; flow rate: 2.5 mL / min, column temperature: 40°C; back pressure: 100 bar.

[0307] SFC method 2: Instrument: CAS-SH-ANA-SFC-L (Waters UPCC with PDA detector); Column: Chiralcel OD-3150×4.6 mm ID, 3 um; Mobile phase: A: CO2, B: methanol (0.05% DEA) Gradient: from 5% to 40% B in 4 min and from 40% to 5% B in 0.2 min, then maintain 5% B for 1.8 min; Flow rate: 2.5 mL / min; Column temperature: 35°C; Back pressure: 1500 psi.

[0308] 6-Piperazin-1-yl-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (P1) and (6R)-6-piperazin-1-yl-2-azaspiro[3.4]octane-2-carboxylate Ethyl spiro[3.4]octane-2-carboxylate (P1A)

[0309]

[0310] Step 1. Synthesis of tert-butyl 6-(4-benzyloxycarbonylpiperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylate (C1): tert-butyl 6-oxo-2-azaspiro[3.4]octane-2-carboxylate (1A, commercially available, 100 g, 444 mmol), benzyl piperazine-1-carboxylate (commercially available, 117 g, 532 mmol) and To a mixture of molecular sieves (67.5 g) in DCE (2.30 L) was added CH3COOH (13.2 g, 221 mmol). The mixture was stirred at 25°C for 60 min, then NaBH(OAc)3 (235 g, 1.10 mol) was added portionwise and stirred at 25°C for 15 hours. The mixture was poured into a saturated aqueous NaHCO3 solution (1.70 L), stirred for 10 min and separated. The aqueous phase was extracted with DCM (3×800 mL). The combined organic phases were washed with brine (1.20 L), dried over anhydrous Na2SO4, filtered and concentrated in vacuo at 50°C. The crude product was triturated with petroleum ether (1.80 L) at 25°C for 60 min. The suspension was filtered and the filter cake was dried in vacuo at 50°C to give tert-butyl 6-(4-benzyloxycarbonylpiperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylate. 1 HNMR (CDCl3400MHz)δ H =7.36-7.33(m,5H),5.12(s,2H),3.86-3.75(m,2H),3.74-3.67(m,2H),3.53(t,J=4.8Hz,4H),2.57(t,J=8.0Hz,1H),2.4 3(s,4H),2.09(dd,J=6.8,12.4Hz,1H),1.96-1.78(m,3H),1.70(dd,J=9.6,12.4Hz,1H),1.60-1.49(m,1H),1.43(s,9H).

[0311] Step 2. Synthesis of benzyl 4-(2-azaspiro[3.4]octan-6-yl)piperazine-1-carboxylate (C2): To a solution of tert-butyl 6-(4-benzyloxycarbonylpiperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylate (170 g, 381 mmol) in DCM (800 mL) was added TFA (286 g, 2.51 mol). The mixture was stirred at 25 ° C for 12 hours. The reaction mixture was concentrated under reduced pressure at 50 ° C to give benzyl 4-(2-azaspiro[3.4]-octan-6-yl)-piperazine-1-carboxylate (340 g, crude, TFA salt), which was used directly in the next step without further purification. 1 H NMR (DMSO-d 6400 MHz) δ H =7.47-7.30(m,5H),5.17-5.05(m,2H),4.25-4.05(m,2H),3.98-3.71(m,4H),3.65-3.53(m,1H),3.51-3.35(m,2H),3.20(d,J= 5.2Hz,2H),3.09-2.86(m,2H),2.35(dd,J=8.4,13.6Hz,1H),2.10-1.96(m,3H),1.85(dd,J=6.0,8.4Hz,1H),1.79-1.69m,1H). LCMS[M+H] + 330.

[0312] Step 3. Synthesis of ethyl 6-(4-benzyloxycarbonylpiperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylate (C3): To a mixture of benzyl 4-(2-azaspiro[3.4]octan-6-yl)piperazine-1-carboxylate (220 g, 496 mmol, TFA salt) in DCM (1.50 L) and H O (750 mL) was added NaHCO (541 g, 6.45 mol) at 25° C. The reaction mixture was stirred at 25° C. for 10 min, and ethyl chloroformate (178 g, 1.64 mol) was added dropwise at 25° C. The mixture was stirred at 25° C. for 1 hour. The reaction mixture was poured into water (500 mL) and stirred at 25° C. for 30 min, and the phases were separated. The aqueous phase was extracted with DCM (2×600 mL). The combined organic phases were washed with brine (500 mL), dried over anhydrous Na2SO4, filtered and concentrated in vacuo to give ethyl 6-(4-benzyloxycarbonylpiperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylate. 1 HNMR (CDCl3400MHz)δ H=7.41-7.27(m,5H),5.12(s,2H),4.08(q,J=7.1Hz,2H),3.90-3.79(m,2H),3.79-3.73(m,2H),3.50(t,J=4.8Hz,4H),2.60-2.49(m,1H) ,2.40(s,4H),2.08(dd,J=7.2,12.8Hz,1H),1.93-1.81(m,3H),1.68(dd,J=9.6,12.8Hz,1H),1.58-1.46(m,1H),1.22(t,J=7.2Hz,3H).

[0313] Step 4. Synthesis of 6-piperazine-1-yl-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (P1): Under N2 atmosphere, to a solution of 6-(4-benzyloxycarbonylpiperazine-1-yl)-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (800 mg, 1.99 mmol) in EtOH (10.0 mL) was added wet Pd / C (80.0 mg, 10 wt %). The suspension was degassed in vacuo and purged with H2 gas several times. The mixture was stirred at 50°C under H2 gas (40 psi) for 16 hours. The reaction mixture was filtered and the filtrate was concentrated to give 6-piperazine-1-yl-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester, which was used directly in the next step. 1 H NMR (CDCl3400MHz)δ H =4.09(q,J=7.2Hz,2H),3.90-3.70(m,4H),2.98-2.82(m,3H),2.65-2.35(m,5H),2.12-2. 05(m,1H),1.95-1.75(m,4H),1.73-1.62(m,1H),1.55-1.46(m,1H),1.23(t,J=7.2Hz,3H).

[0314] Step 5. Synthesis of ethyl (6R)-6-(4-benzyloxycarbonylpiperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylate (C3A) and ethyl (6S)-6-(4-benzyloxycarbonylpiperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylate (C3B): A racemic mixture of ethyl 6-(4-benzyloxycarbonylpiperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylate (430 g, 1.07 mol) was purified by SFC (column: DAICEL CHIRALCEL OJ (250mm*50mm, 10um); mobile phase: [0.1% NH3H2OMeOH]; B%: 20%-20%, min) to obtain (6R)-6-(4-benzyloxycarbonylpiperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester and (6S)-6-(4-benzyloxycarbonylpiperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester with a purity of 95.9%. LCMS [M+H] + 402.

[0315] Step 6. Synthesis of (6R)-6-piperazin-1-yl-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (P1A): To a solution of (6R)-6-(4-benzyloxycarbonylpiperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (120 g, 299 mmol) in MeOH (1200 mL) was added wet 10% Pd / C (15.0 g) at 20°C. The suspension was degassed under vacuum and purged with H2 gas several times. The mixture was stirred at 25°C under H2 gas (15 psi) for 16 hours. The reaction mixture was filtered and washed with MeOH (2×1,000 mL). The filtrate was concentrated to give (6R)-6-piperazin-1-yl-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. 1 HNMR (CDCl3400MHz)δ H =4.08(q,J=7.2Hz,2H),3.92-3.71(m,4H),2.88(t,J=5.2Hz,4H),2.58-2.33(m,5H),2.15-2 .03(m,1H),1.97-1.75(m,3H),1.71-1.63(m,1H),1.57-1.43(m,1H),1.22(t,J=7.2Hz,3H). LCMS[M+H] + 268.

[0316] (6R)-6-[4-(3-Bromo-5-fluoro-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (P2)

[0317]

[0318] A mixture of (6R)-6-piperazin-1-yl-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (1.27 g, 4.75 mmol), 3-bromo-2,5-difluoro-pyridine (921 mg, 4.75 mmol) and DIPEA (14.0 mL, 80.2 mmol) in pyridine (10.0 mL) was degassed and purged with N2 gas three times and stirred at 120 ° C for 12 hours under N2 gas atmosphere. The reaction mixture was concentrated under reduced pressure to remove volatile reagents. The residue was purified by flash chromatography on silica gel (eluent: 0 to 10% MeOH / DCM) to give (6R)-6-[4-(3-bromo-5-fluoro-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. LCMSm / z[M+H] + 443.

[0319] (6R)-6-[4-(3-Bromo-2-pyridyl)piperazin-1-yl]-2-azaspiro[3.4]-octane-2-carboxylic acid ethyl ester (P3)

[0320]

[0321] At 20 ℃, to a mixture of 3-bromo-2-fluoro-pyridine (1.65 g, 9.35 mmol) and (6R)-6-piperazine-1-yl-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (2.50 g, 9.35 mmol) in DMA (25.0 mL) was added K CO (2.58 g, 18.7 mmol) in one portion. The mixture was heated to 140 ℃ and stirred for 12 hours. The mixture was cooled to 20 ℃ and concentrated under reduced pressure at 50 ℃. Water (50 mL) was added to the remaining residue and extracted with DCM (4×50 mL). The combined organic phases were washed with brine (150 mL), dried over anhydrous Na SO , filtered and concentrated under reduced pressure at 50 ℃. The residue was purified by silica gel chromatography (eluent: 0 to 5% MeOH / DCM) to afford (6R)-ethyl 6-[4-(3-bromo-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate. 1 H NMR (CDCl3400MHz)δ H=8.22(dd,J=1.6Hz,4.8Hz,1H),8.77(dd,J=1.6Hz,8.0Hz,1H),6.76(dd,J=4.8Hz,7.6Hz,1H),5.30(s,1H),4.10(q,J=7.2Hz,2H),3.95-3.74(m,4H), 3.43-3.27(m,4H),2.68-2.60(m,4H),2.14(dd,J=7.2Hz,12.8Hz,1H),1.99 -1.88(m,2H),1.88-1.71(m,2H),1.60-1.52(m,1H),1.24(t,J=7.2Hz,3H). LCMS[M+H] + 423,425.

[0322] 6-(4-(3-(Pyrazin-2-yl)pyridin-2-yl)piperazin-1-yl)-2-azaspiro[3.4]octane (P4)

[0323]

[0324] Step 1: Synthesis of 2-(2-fluoropyridin-3-yl)pyrazine (C4): To a mixture of 2-chloropyrazine (1.56 mL, 17.5 mmol), (2-fluoro-3-pyridinyl)boronic acid (2.46 g, 17.5 mmol), and NaCO (5.55 g, 52.4 mmol) in 1,4-dioxane (30.0 mL) and HO (6.00 mL) was added Pd(dppf)Cl (1.28 g, 1.75 mmol) in one portion at 20°C and degassed several times under N. The mixture was stirred at 100°C for 16 hours. The reaction mixture was concentrated under reduced pressure. The residue was diluted with EtOAc (100 mL), and the organic layer was washed with water (2 x 50 mL), brine (50 mL), dried over anhydrous NaSO, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by flash chromatography on silica gel (0 to 30% EtOAc / petroleum ether as eluent) to give 2-(2-fluoro-3-pyridyl)pyrazine. LCMS [M+H] + 176.

[0325] Step 2: Synthesis of tert-butyl 6-(4-(3-(pyrazin-2-yl)pyridin-2-yl)piperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylate (C5): A solution of 2-(2-fluoro-3-pyridinyl)pyrazine (3.26 g, 18.6 mmol) and tert-butyl 6-piperazin-1-yl-2-azaspiro[3.4]octane-2-carboxylate (5.50 g, 18.6 mmol) in DIPEA (10.0 mL) was stirred at 140° C. for 16 hours. The mixture was concentrated under reduced pressure to give a residue. The residue was diluted with DCM (200 mL) and washed with water (2×100 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 , filtered and concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel (eluent: 0 to 10% MeOH / DCM) to give tert-butyl 6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate and by preparative HPLC (column: Phenomenex Gemini-NX 80×40 mm×3 um; mobile phase: [water (0.05% NH3H2O+10 mM NH4HCO3)-ACN]; B%: 35%-65%, 8 min) to give tert-butyl 6-[4-(3-pyrazin-2-yl-2-pyridinyl)-piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate. 1 H NMR (CDCl3400MHz)δ H =9.24(s,1H),8.65(s,1H),8.47(d,J=2.4Hz,1H),8.32(dd,J=2.0Hz,4.8Hz,1H),7.86(dd,J=1.6Hz,7.2Hz,1H),7.01(dd,J=4.8Hz,7.6Hz,1H),3.82 -3.68(m,4H),3.15(s,4H),2.60-2.39(m,5H),2.06(dd,J=6.8Hz,12.4HZ, 1H),1.93-1.75(m,3H),1.66-1.57(m,1H),1.56-1.47(m,1H),1.42(s,9H). LCMS[M+H] + 451.

[0326] Step 3: Synthesis of (6R)-tert-butyl 6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (C5A) and (6S)-tert-butyl 6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (C5B): tert-Butyl 6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (2.68 g, 5.95 mmol) was purified by preparative SFC. The sample was purified by SFC (column: DAICELCHIRALCEL OD (250 mm×50 mm, 10 μm); mobile phase: [0.1% NH 3 H 2 O ETOH]; B%: 30%-30%, 7 min) to give Peak 1 (6R)-6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid tert-butyl ester (1.18 g, 2.61 mmol, 43.8% yield, 99.5% purity) and Peak 2 (6S)-6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid tert-butyl ester as an off-white solid. Peak 1: LCMS [M+H] + 451. Peak 2: LCMS [M+H] + 451.

[0327] Step 4: Synthesis of 6-(4-(3-(pyrazin-2-yl)pyridin-2-yl)piperazin-1-yl)-2-azaspiro[3.4]octane (P4): To a solution of tert-butyl 6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]-octane-2-carboxylate (6.30 g, 14.0 mmol) in DCM (30.0 mL) was added TFA (19.6 mL, 264 mmol). The mixture was stirred at 20° C. for 5 hours. The mixture was adjusted to pH=9 with 10% aqueous NaOH and then diluted with DCM (50 mL). The aqueous phase was extracted with DCM (2×30 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 and filtered. The organic layer was concentrated under reduced pressure to obtain 6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane. 1 H NMR (CDCl3400MHz)δ H=9.23(d,J=1.2Hz,1H),8.73-8.57(m,1H),8.46(d,J=2.0Hz,1H),8.39-8.2 7(m,1H),7.86(d,J=7.2Hz,1H),7.08-6.92(m,1H),3.72-3.38(m,2H),3.21 -3.00(m,7H),2.45(d,J=2.4Hz,5H),2.24-2.15(m,1H),2.10-2.02(m,1H), 1.96-1.88(m,1H),1.77-1.68(m,1H),1.65-1.54(m,1H),1.48-1.36(m,1H). LCMS[M+H] + 351.

[0328] Ethyl 6-[4-(3-hydroxy-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (P5)

[0329]

[0330] Step 1: Synthesis of tert-butyl 4-(3-hydroxypyridin-2-yl)piperazine-1-carboxylate (C6): A mixture of 2-fluoropyridin-3-ol (2.00 g, 17.7 mmol), tert-butyl piperazine-1-carboxylate (6.59 g, 35.4 mmol), and DIPEA (9.24 mL, 53.1 mmol) was degassed and purged with N2 gas three times. The mixture was then heated to 140°C and stirred at 140°C under an atmosphere of N2 gas for 16 hours. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by flash chromatography on silica gel (eluent: 0 to 20% EtOAc / petroleum ether) to give tert-butyl 4-(3-hydroxy-2-pyridinyl)piperazine-1-carboxylate. 1 H NMR (CDCl3400MHz) δH=7.96(dd,J=1.6Hz,4.8Hz,1H),7.22(dd,J=1.6Hz,8.0Hz,1H) ,7.03(dd,J=4.8Hz,8.0Hz,1H),3.67-3.59(m,4H),3.09-2.98(m,4H),1.50(s,9H). LCMS[M+H]+280.

[0331] Step 2: Synthesis of 2-(piperazin-1-yl)pyridin-3-ol (C7): To a solution of tert-butyl 4-(3-hydroxy-2-pyridinyl)piperazine-1-carboxylate (3.00 g, 10.7 mmol) in MeOH (10.0 mL) was added HCl / MeOH (4 M, 15.0 mL). The mixture was stirred at 20° C. for 1.5 hours. The reaction mixture was concentrated under reduced pressure to give crude 2-(piperazin-1-yl)pyridin-3-ol. LCMS [M+H] 180.

[0332] Step 3: Synthesis of tert-butyl 6-(4-(3-hydroxypyridin-2-yl)piperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylate (C8): To a solution of 2-piperazin-1-ylpyridin-3-ol (1.90 g, 10.6 mmol, HCl salt) in DCE (20.0 mL) was added ethyl 6-oxo-2-azaspiro[3.4]octane-2-carboxylate (2.09 g, 10.6 mmol), NaBH(OAc) (2.70 g, 12.7 mmol) and CHCOOH (318 mg, 5.30 mmol) and EtN (4.43 mL, 31.8 mmol). The mixture was stirred at 25° C. for 16 hours. Saturated aqueous NaHCO (20 mL) was added to the reaction mixture and separated. The aqueous phase was extracted with EtOAc (3 x 150 mL), dried over anhydrous NaSO, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by flash chromatography on silica gel (eluent: 0 to 10% MeOH / DCM) to give tert-butyl 6-[4-(3-hydroxy-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate. 1 HNMR (CDCl3400MHz) δH=7.95(dd,J=1.6Hz,4.8Hz,1H),7.19(dd,J=1.6Hz,8.0Hz,1H),6.99(dd,J=3.2Hz,4.4Hz,1H),3.89-3.80(m,2H ),3.79-3.74(m,2H),3.13(t,J=4.8Hz,4H),2.77-2.60(m,5H),2.20-2.10(m,1H),2.00-1.75(m,4H),1.65-1.55(m,1H),1.46(s,9H). LCMS[M+H]+389.

[0333] Step 4: Synthesis of 2-(4-(2-azaspiro[3.4]octan-6-yl)piperazin-1-yl)pyridin-3-ol (C9): To a solution of tert-butyl 6-[4-(3-hydroxy-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]-octane-2-carboxylate (2.00 g, 5.15 mmol) in DCM (50.0 mL) was added TFA (5.72 mL, 77.2 mmol). The mixture was stirred at 20° C. for 16 hours. The reaction mixture was concentrated under reduced pressure to give 2-[4-(2-azaspiro[3.4]octan-6-yl)piperazin-1-yl]pyridin-3-ol. LCMS [M+H]+ 289.

[0334] Step 5: Synthesis of ethyl 6-(4-(3-((ethoxycarbonyl)oxy)pyridin-2-yl)piperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylate (C10): To a solution of 2-[4-(2-azaspiro[3.4]octan-6-yl)piperazin-1-yl]pyridin-3-ol (1.60 g, 5.55 mmol) in DCM (10.0 mL) at 0° C. was added EtN (1.68 g, 16.6 mmol) and ethyl chloroformate (2.80 g, 25.8 mmol). The mixture was stirred at 0° C. for 2 hours. The reaction mixture was quenched by the addition of water (10 mL) at 0° C. and then diluted with DCM (5 mL). The mixture was extracted with DCM (3×10 mL). The combined organic layers were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give ethyl 6-[4-(3-ethoxycarbonyloxy-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate. LCMS [M+H]+ 433.

[0335] Step 6: Synthesis of ethyl 6-[4-(3-hydroxy-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (P5): A mixture of ethyl 6-[4-(3-ethoxycarbonyloxy-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (2.50 g, 5.78 mmol) and NaOH (1.20 g, 30.0 mmol) in EtOH (20.0 mL) and H O (10.0 mL) was heated to 60° C. and stirred at 60° C. for 16 hours. The mixture was cooled to 25° C. and concentrated in vacuo. DCM (200 mL) was then added to the mixture and filtered. The filtrate was then concentrated in vacuo. The residue was purified by flash chromatography on silica gel (0 to 10% MeOH / DCM as eluent) to give ethyl 6-[4-(3-hydroxy-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate. LCMS [M+H]+361.

[0336] 2-(5-Fluoro-2-piperazin-1-yl-3-pyridyl)-1,3,4-thiadiazole (P6)

[0337]

[0338] Step 1: Synthesis of tert-butyl 4-(5-fluoro-3-methoxycarbonyl-2-pyridinyl)piperazine-1-carboxylate (C11): A mixture of methyl 2-chloro-5-fluoro-pyridine-3-carboxylate (1.50 g, 7.91 mmol), tert-butyl piperazine-1-carboxylate (1.77 g, 9.50 mmol), and KCO (2.19 g, 15.8 mmol) in DMF (15.0 mL) was degassed at 25° C. and purged with N gas several times. The mixture was then heated to 120° C. and stirred at 120° C. under an atmosphere of N gas for 12 hours. The mixture was poured into H O (50 mL). The mixture was extracted with EtOAc (3×30 mL). The combined organic layers were washed with H O (100 mL), brine (100 mL), dried over anhydrous Na SO , filtered, and concentrated in vacuo to yield a residue. The residue was purified by flash chromatography on silica gel (eluent: 0 to 7% EtOAc / petroleum ether) to give tert-butyl 4-(5-fluoro-3-methoxycarbonyl-2-pyridinyl)piperazine-1-carboxylate. LCMS [M+H] + 340.

[0339] Step 2: Synthesis of tert-butyl 4-[5-fluoro-3-(hydrazinecarbonyl)-2-pyridinyl]piperazine-1-carboxylate (C12): To a solution of tert-butyl 4-(5-fluoro-3-methoxycarbonyl-2-pyridinyl)piperazine-1-carboxylate (1.81 g, 5.33 mmol) in EtOH (18.0 mL) was added NH2NH2·H2O (4.71 g, 80.0 mmol, 85.0% purity) at 25°C. The mixture was then heated to 80°C and stirred at 80°C for 16 hours. The mixture was concentrated in vacuo to give a residue. The residue was purified by trituration with EtOAc / petroleum ether (1 / 1, 15 mL) at 25°C and filtration. The filtrate was concentrated under reduced pressure to give tert-butyl 4-[5-fluoro-3-(hydrazinecarbonyl)-2-pyridinyl]piperazine-1-carboxylate. 1 H NMR (CDCl3400MHz)δ H =10.10(br s,1H),8.25(d,J=2.8Hz,1H),8.10(dd,J=2.8Hz,8.4Hz,1H),3.66-3.56(m,4H),3.13-3.02(m,4H),1.51-1.45(m,9H). LCMS[M+H] + 340.

[0340] Step 3: Synthesis of tert-butyl 4-[5-fluoro-3-(formylaminocarbamoyl)-2-pyridinyl]piperazine-1-carboxylate (C13): To a solution of tert-butyl 4-[5-fluoro-3-(hydrazinecarbonyl)-2-pyridinyl]piperazine-1-carboxylate (1.30 g, 3.83 mmol) in toluene (26.0 mL) was added HCO₂H (1.88 g, 38.3 mmol, 98% purity) at 25° C. under an N₂ atmosphere. The mixture was then heated to 50° C. and stirred at 50° C. for 1 hour. The mixture was adjusted to pH=8 with saturated aqueous NaHCO₃. The mixture was extracted with EtOAc (3×50 mL). The combined organic phases were washed with brine (150 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated in vacuo to give a residue. The residue was purified by flash chromatography on silica gel (eluent: 0 to 35% EtOAc in petroleum ether) to give 4-[5-fluoro-3-(formylaminocarbamoyl)-2-pyridinyl]-piperazine-1-carboxylic acid tert-butyl ester. 1 HNMR (CDCl3400MHz)δ H =13.15(br d,J=7.2Hz,1H),9.04(br d,J=6.4Hz,1H),8.36(d,J=2.0Hz,1H),8.29-8.11(m,2H),3.76(br s,4H),3.08(br s,4H),1.48(s,9H). LCMS[M+H] + 368.

[0341] Step 4: Synthesis of tert-butyl 4-[5-fluoro-3-(1,3,4-thiadiazol-2-yl)-2-pyridinyl]piperazine-1-carboxylate (C14): To a solution of tert-butyl 4-[5-fluoro-3-(formylaminocarbamoyl)-2-pyridinyl]piperazine-1-carboxylate (1.00 g, 2.72 mmol) in toluene (50.0 mL) was added Lawesson's reagent (1.32 g, 3.27 mmol) at 25°C under an N2 atmosphere. The mixture was then heated to 90°C and stirred at 90°C for 16 hours. Saturated aqueous NaHCO3 solution (60 mL) was added to the mixture and stirred for 5 minutes. The mixture was extracted with EtOAc (3×50 mL). The combined organic phases were washed with brine (150 mL), dried over anhydrous Na2SO4, filtered, and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (eluent: 0 to 25% EtOAc in petroleum ether) to give tert-butyl 4-[5-fluoro-3-(1,3,4-thiadiazol-2-yl)-2-pyridinyl]piperazine-1-carboxylate. 1 H NMR (CDCl3400MHz)δ H=9.28-9.13(m,1H),8.39(dd,J=2.8Hz,8.4Hz,1H),8.34(d,J=2.8Hz,1H),3.67-3.58(m,4H),3.09-2.96(m,4H),1.48(s,9H). LCMS[M+H] + 366.

[0342] Step 5: Synthesis of 2-(5-fluoro-2-piperazin-1-yl-3-pyridinyl)-1,3,4-thiadiazole (P6): Under a nitrogen atmosphere, HCl / EtOAc (4.00 M, 2.00 mL) was added to a solution of tert-butyl 4-[5-fluoro-3-(1,3,4-thiadiazole-2-yl)-2-pyridinyl]piperazine-1-carboxylate (100 mg, 274 umol) in DCM (3.00 mL) at 25°C. The mixture was then stirred at 25°C for 12 hours. The mixture was adjusted to pH = 8 with saturated aqueous NaHCO3 and extracted with EtOAc (3×20 mL). The combined organic phases were washed with brine (50 mL), dried over anhydrous Na2SO4, filtered and concentrated in vacuo to give a solid. 2-(5-Fluoro-2-piperazin-1-yl-3-pyridinyl)-1,3,4-thiadiazole was used in the next step without further purification. LCMS [M+H] + 266.

[0343] Ethyl 6-(4-(3-bromopyridin-2-yl)piperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylate (P7)

[0344]

[0345] Step 1: Synthesis of 1-(3-bromopyridin-2-yl)piperazine (C15): A mixture of 3-bromo-2-chloro-pyridine (1.00 g, 5.20 mmol) and piperazine (537 mg, 6.24 mmol) in DMSO (20.0 mL) was stirred at 100° C. for 16 hours. The mixture was worked up with another product of the same scale. The residue was poured into water (150 mL). The aqueous phase was extracted with EtOAc (3×50 mL). The combined organic phases were washed with brine (100 mL), dried over anhydrous Na 2 SO 4 , filtered, and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (eluent: 0 to 30% EtOAc / MeOH) to give 1-(3-bromo-2-pyridinyl)piperazine. 1 HNMR (CDCl3400MHz)δ H=8.23(dd,J=1.6Hz,4.8Hz,1H),7.79(dd,J=1.6Hz,7.6Hz,1H),6.78(dd,J=4.8Hz,7.6Hz,1H),3.35-3.27(m,4H),3.10-3.01(m,4H). LCMS[M+H] + 242,244.

[0346] Step 2: Synthesis of tert-butyl 6-(4-(3-bromopyridin-2-yl)piperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylate (C16): To a mixture of tert-butyl 6-oxo-2-azaspiro[3.4]octane-2-carboxylate (409 mg, 1.82 mmol) and 1-(3-bromo-2-pyridinyl)piperazine (550 mg, 1.82 mmol) in DCE (10.0 mL) were added HOAc (0.100 mL) and NaBH(OAc) (1.16 g, 5.45 mmol) in one portion. The mixture was stirred at 20° C. for 16 hours. The residue was poured into saturated aqueous NaHCO (50 mL). The aqueous phase was extracted with DCM (3×30 mL). The combined organic phases were washed with brine (50 mL), dried over anhydrous NaSO, filtered and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (eluent: 0 to 15% MeOH / DCM) to afford tert-butyl 6-[4-(3-bromo-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]-octane-2-carboxylate. 1 H NMR (CDCl3400MHz)δ H =8.23(dd,J=1.6Hz,4.8Hz,1H),7.78(dd,J=1.6Hz,7.6Hz,1H),6.77(dd,J=4.8H z,7.6Hz,1H),3.89-3.83(m,1H),3.83-3.78(m,1H),3.77-3.71(m,2H),3.39(br s,4H),2.65(br s,5H),2.20-2.10(m,1H),1.99-1.81(m,3H),1.79-1.70(m,1H),1.45(s,9H). LCMS[M+H] + 451,453.

[0347] Step 3: Synthesis of 6-(4-(3-bromopyridin-2-yl)piperazin-1-yl)-2-azaspiro[3.4]octane (C17): To a mixture of tert-butyl 6-[4-(3-bromo-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]-octane-2-carboxylate (850 mg, 1.88 mmol) in 1,4-dioxane (10.0 mL) was added HCl / 1,4-dioxane (4 M, 4.00 mL) in one portion over 1 hour at 20° C. The mixture was concentrated to give 6-[4-(3-bromo-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane, which was used directly in the next step.

[0348] Step 4: Synthesis of ethyl 6-(4-(3-bromopyridin-2-yl)piperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylate (P7): To a mixture of 6-[4-(3-bromo-2-pyridin-2-yl)piperazin-1-yl]-2-azaspiro[3.4]octane (1.00 g, 2.85 mmol, HCl salt) and EtN (1.44 g, 14.2 mmol) in DCM (10.0 mL) at 0° C. under a nitrogen atmosphere was added ethyl chloroformate (970 mg, 8.94 mmol) in one portion. The mixture was stirred at 20° C. for 2 hours. The residue was poured into water (50 mL). The aqueous phase was extracted with DCM (3×30 mL). The combined organic phases were washed with brine (50 mL), dried over anhydrous NaSO, filtered and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (eluent: 0 to 20% MeOH / DCM) to afford ethyl 6-[4-(3-bromo-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate. 1 HNMR (CDCl3400MHz)δ H =8.23(dd,J=1.6Hz,4.8Hz,1H),7.78(dd,J=1.6Hz,7.6Hz,1H),6.77(dd,J=4.8Hz,7.6Hz, 1H),4.16-4.10(m,2H),3.94-3.89(m,1H),3.88-3.83(m,1H),3.83-3.77(m,2H),3.38(br s,4H),2.64(br s,5H),2.20-2.10(m,1H),1.99-1.90(m,2H),1.89-1.73(m,2H),1.61-1.54(m,1H),1.30-1.24(m,3H). LCMS[M+H] + 423,425.

[0349] 6-[4-[3-(2,6-dimethylphenyl)-2-pyridyl]piperazin-1-yl]-2-azaspiro[3.4]-octane (P8)

[0350]

[0351] Step 1: Synthesis of tert-butyl 4-[3-(2,6-dimethylphenyl)-2-pyridinyl]piperazine-1-carboxylate (C18): A mixture of tert-butyl 4-[3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-pyridinyl]-piperazine-1-carboxylate (200 mg, 514 umol), 2-bromo-1,3-dimethyl-benzene (114 mg, 617 umol), Pd(t-Bu3P)2 (26.3 mg, 51.4 umol), Cs2CO3 (502 mg, 1.54 mmol), toluene (4.50 mL) and H2O (0.500 mL) was added to a sealed tube under N2 in a glove box, then heated to 80°C and stirred at 80°C for 16 hours. The mixture was then cooled to 25°C and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (0 to 8% EtOAc / petroleum ether as eluent) to give tert-butyl 4-[3-(2,6-dimethylphenyl)-2-pyridinyl]-piperazine-1-carboxylate. LCMS [M+H]+368.

[0352] Step 2: Synthesis of 1-[3-(2,6-dimethylphenyl)-2-pyridyl]piperazine (C19): At 20° C., a mixture of tert-butyl 4-[3-(2,6-dimethylphenyl)-2-pyridyl]piperazine-1-carboxylate (110 mg, 299 umol) in EtOAc (5.00 mL) was added to the mixture, and then HCl / EtOAc (4 M, 3.00 mL) was added to the mixture. The mixture was stirred at 20° C. for 4 hours. The mixture was concentrated in vacuo and used in the next step without further purification. LCMS [M+H] + 268.

[0353] Step 3: Synthesis of tert-butyl 6-[4-[3-(2,6-dimethylphenyl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (C20): A mixture of 1-[3-(2,6-dimethylphenyl)-2-pyridinyl]piperazine (90.0 mg, 296 umol, HCl salt) and Et3N (150 mg, 1.48 mmol) in DCE (5.00 mL) was stirred at 20° C. for 10 min. Then, tert-butyl 6-oxo-2-azaspiro[3.4]octane-2-carboxylate (66.7 mg, 296 umol) was added to the mixture and stirred at 20° C. for 20 min. AcOH (12.8 mg, 214 umol) and NaBH (OAc) 3 (188 mg, 889 umol) were then added to the mixture at 20 ° C and stirred at 20 ° C for 16 hours. Then, a NaHCO 3 aqueous solution (10%, 25 mL) was added to the mixture, and the mixture was extracted with DCM (50 mL × 2) and dried over anhydrous Na 2 SO 4. The combined organic phases were concentrated in vacuo. The residue was purified by flash chromatography on silica gel (eluent: 0 to 3% MeOH / DCM) to give tert-butyl 6- [4- [3- (2,6-dimethylphenyl) -2-pyridyl] piperazin-1-yl] -2- azaspiro [3.4] octane-2-carboxylate. LCMS [M + H] + 477.

[0354] Step 4: Synthesis of 6-[4-[3-(2,6-dimethylphenyl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane (P8): tert-Butyl 6-[4-[3-(2,6-dimethylphenyl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (95.0 mg, 199 umol) was dissolved in DCM (5.00 mL) to obtain a clear solution. TFA (0.250 mL, 3.39 mmol) was then added to the mixture and stirred at 20° C. for 2 hours. The mixture was concentrated in vacuo and used in the next step without further purification. LCMS [M+H] 377.

[0355] (6R)-6-[4-[3-[(E)-3-(dimethylamino)prop-2-enoyl]-2-pyridinyl]-piperazin-1-yl]-2- Ethyl azaspiro[3.4]octane-2-carboxylate (P9)

[0356]

[0357] Step 1: Synthesis of ethyl 6-[4-(3-acetyl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (C21): To a solution of ethyl 6-piperazin-1-yl-2-azaspiro[3.4]octane-2-carboxylate (300 mg, 1.12 mmol) in DIPEA (10.0 mL) was added 1-(2-chloro-3-pyridinyl)ethanone (175 mg, 1.12 mmol) in one portion at 20°C. The mixture was heated to 130°C and stirred for 16 hours. The mixture was cooled to 20°C. Water (20 mL) and DCM (10 mL) were added to the mixture and stirred for 5 minutes and separated. The aqueous phase was extracted with DCM (2×10 mL). The combined organic phases were concentrated under reduced pressure at 50°C. The residue was purified by flash chromatography on silica gel (eluent: 0 to 5% MeOH / DCM) to afford ethyl 6-[4-(3-acetyl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate. 1 H NMR (CDCl3400MHz)δ H =8.29(dd,J=2.0Hz,4.8Hz,1H),7.72(dd,J=1.6Hz,7.6Hz,1H),6.83(dd,J=4.8Hz,7.6Hz,1H),4.10(q,J=7.2Hz,2H),3.92-3.74(m,4H), 3.42-3.30(m,4H),2.70-2.45(m,8H),2.15-2.10(m,1H),1.98-1.80(m,3H),1.77-1.69(m,1H),1.55-1.53(m,1H),1.24(t,J=7.2Hz,3H). LCMS[M+H] + 387.

[0358] Step 2: Synthesis of (6R)-6-[4-[3-[(E)-3-(dimethylamino)prop-2-enoyl]-2-pyridinyl]-piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (P9): A mixture of (6R)-6-[4-(3-acetyl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (120 mg, 310 umol) and 1,1-dimethoxy-N,N-dimethyl-methylamine (1.20 mL, 9.03 mmol) was heated to 100° C. and stirred at 100° C. for 16 hours. The mixture was concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel (eluent: 0 to 8% MeOH / DCM) to give (6R)-ethyl 6-[4-[3-[(E)-3-(dimethylamino)prop-2-enoyl]-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate. LCMS [M+H]+ 442.

[0359] 6-[4-(5-chloro-3-pyrazin-2-yl-2-pyridinyl)-2,2,3,3,5,5,6,6-octadeuterated-piperazin-1-yl]-2- Ethyl azaspiro[3.4]octane-2-carboxylate (P10)

[0360]

[0361] Step 1: Synthesis of tert-butyl 4-(5-chloro-3-pyrazin-2-yl-2-pyridinyl)-2,2,3,3,5,5,6,6-octadeuterio-piperazine-1-carboxylate (C22): A mixture of 2-(5-chloro-2-fluoro-3-pyridinyl)pyrazine (450 mg, 2.15 mmol) and tert-butyl 2,2,3,3,5,5,6,6-octadeuterio-piperazine-1-carboxylate (417 mg, 2.15 mmol) in pyridine (10.0 mL) and DIPEA (10.0 mL) was heated to 130° C. and stirred at 130° C. for 12 hours. The reaction mixture was poured into water (50 mL) at 20° C. and extracted with EtOAc (3×50 mL). The combined organic layers were washed with brine (100 mL), dried over Na 2 SO 4 , filtered, and concentrated under reduced pressure to give a residue. The residue was purified by flash chromatography on silica gel (eluent: 0 to 70% EtOAc / petroleum ether) to give 4-(5-chloro-3-pyrazin-2-yl-2-pyridinyl)-2,2,3,3,5,5,6,6-octadeutero-piperazine-1-carboxylic acid tert-butyl ester. LCMS [M+H] + 384.

[0362] Step 2: Synthesis of 1-(5-chloro-3-pyrazin-2-yl-2-pyridinyl)-2,2,3,3,5,5,6,6-octadeutero-piperazine (C23): To a solution of tert-butyl 4-(5-chloro-3-pyrazin-2-yl-2-pyridinyl)-2,2,3,3,5,5,6,6-octadeutero-piperazine-1-carboxylate (600 mg, 1.56 mmol) in 1,4-dioxane (10.0 mL) was added HCl / EtOAc (4 M, 4.00 mL) in one portion at 20°C. The mixture was stirred at 20°C for 1 hour. The mixture was concentrated under reduced pressure to afford 1-(5-chloro-3-pyrazin-2-yl-2-pyridinyl)-2,2,3,3,5,5,6,6-octadeutero-piperazine. The product was used directly in the next step without further purification. LCMS [M+H] + 284.

[0363] Step 3: Synthesis of 6-[4-(5-chloro-3-pyrazin-2-yl-2-pyridinyl)-2,2,3,3,5,5,6,6-octadeutero-piperazine-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (P10): To a solution of 1-(5-chloro-3-pyrazin-2-yl-2-pyridinyl)-2,2,3,3,5,5,6,6-octadeutero-piperazine (440 mg, 1.55 mmol) and 6-oxo-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (367 mg, 1.86 mmol) in DCE (5.00 mL) was added HOAc (9.31 mg, 155 umol) in one portion under N2 at 0°C. The mixture was stirred at 0°C for 30 min, and then NaBH(OAc)3 (986 mg, 4.65 mmol) was added at 0°C. The mixture was warmed to 20°C and stirred at 20°C for 16 hours. The reaction mixture was poured into water (50 mL) at 20°C and then extracted with EtOAc (3×50 mL). The combined organic layers were washed with brine (50 mL), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel (eluent: 0 to 30% MeOH in DCM) to give 6-[4-(5-chloro-3-pyrazin-2-yl-2-pyridinyl)-2,2,3,3,5,5,6,6-octadeuterated-piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. LCMS[M+H] + 465.

[0364] 3-(2-fluoropyridin-3-yl)isoxazole (P11) and 3-(2-fluoropyridin-3-yl)-4-methylisoxazole (P12)

[0365]

[0366] Step 1: Synthesis of (E)-2-fluoronicotinaldehyde oxime (C24): To a solution of 2-fluoropyridine-3-carbaldehyde (1.00 g, 7.99 mmol) in MeOH (2.00 mL) was added a solution of NH2OH·HCl (833 mg, 12.0 mmol) and KOAc (1.18 g, 12.0 mmol) in H2O (7.00 mL). The mixture was stirred at 15°C for 4 hours. The mixture was concentrated under reduced pressure to remove MeOH. Saturated aqueous Na2CO3 (50 mL) was then added to the residue and extracted with DCM (2×50 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by flash chromatography on silica gel (eluent: 0 to 20% EtOAc / petroleum ether) to give (3E)-2-fluoropyridine-3-carbaldehyde oxime. LCMS [M+H] + 141.

[0367] Step 2: Synthesis of (Z)-2-Fluoro-N-hydroxynicotinimide chloride (C25): (3E)-2-Fluoropyridine-3-carbaldehyde oxime (900 mg, 6.42 mmol) was dissolved in MeCN (15.0 mL). A solution of NCS (1.03 g, 7.71 mmol) in MeCN (15.0 mL) was then slowly added dropwise at 15°C. After addition, the reaction mixture was stirred at 15°C for 3 hours. The mixture was used in the next step without post-treatment.

[0368] Step 3: Synthesis of 3-(2-fluoropyridin-3-yl)-5-(trimethylsilyl)isoxazole (C26): To a solution of (3Z)-2-fluoro-N-hydroxy-pyridine-3-carboximidoyl chloride (1.10 g, 6.30 mmol) and ethynyl(trimethyl)silane (1.05 mL, 7.56 mmol) in MeCN (30.0 mL) was added TEA (965 uL, 6.93 mmol) and the reaction mixture was stirred at 15 ° C for 16 hours. The mixture was concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel (eluent: 0 to 30% EtOAc / petroleum ether) to give [3-(2-fluoro-3-pyridinyl)isoxazol-5-yl]-trimethyl-silane. 1 H NMR (CDCl3400MHz)δ H =8.60-8.41(m,1H),8.30-8.25(m,1H),7.32-7.28(m,1H),6.91-6.88(m,1H),0.38(m,9H). 19 FNMR (CDCl3400MHz)δ F =-68.19. LCMS [M+H] + 237.

[0369] Step 4: Synthesis of 3-(2-fluoropyridin-3-yl)isoxazole (P11): To a solution of [3-(2-fluoro-3-pyridinyl)isoxazol-5-yl]-trimethyl-silane (100 mg, 423 umol) in EtOH (1.00 mL) and MeCN (3.00 mL) was added CsF (46.8 uL, 1.27 mmol). The reaction was stirred at 20 ° C for 2 hours. The mixture was concentrated under reduced pressure. The residue was diluted with DCM (30 mL) and washed with water (20 mL). The organic phase was concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel (eluent: 0 to 50% EtOAc / petroleum ether) to give 3-(2-fluoro-3-pyridinyl)-isoxazole. LCMS [M+H] + 165.

[0370] Step 5: Synthesis of (E)-3-methoxy-3-oxoprop-1-en-1-yl 4-nitrobenzoate (C27): 4-nitrobenzoic acid (3.00 g, 17.9 mmol) and methyl prop-2-ynoate (1.64 mL, 19.7 mmol) were dissolved in MeCN (30.0 mL) and N-methylmorpholine (990 uL, 8.98 mmol) was added. The mixture was then stirred at 40 ° C for 16 hours. The mixture was concentrated under reduced pressure to remove MeCN. The residue was stirred in MTBE (100 mL) at 20 ° C for 30 min, and the mixture was then filtered. The collected filter cake was dried under reduced pressure to give [(E)-3-methoxy-3-oxo-prop-1-enyl] 4-nitrobenzoate. 1 H NMR (CDCl3400MHz)δ H =8.52(d,J=12.8Hz,1H),8.37-8.30(m,4H),5.98(d,J=12.4Hz,1H),3.80(s,3H).

[0371] Step 6: Synthesis of methyl 3-(2-fluoropyridin-3-yl)isoxazole-4-carboxylate (C28): To a solution of (3Z)-2-fluoro-N-hydroxy-pyridine-3-carboximidoyl chloride (1.25 g, 7.16 mmol) in MeCN was added a solution of [(E)-3-methoxy-3-oxo-prop-1-enyl] 4-nitrobenzoate (1.05 mL, 7.88 mmol) in DCM (20.0 mL). TEA (1.10 mL, 7.88 mmol) was then added in one portion. The mixture was stirred at 15 °C for 16 hours. The mixture was concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel (eluent: 0 to 30% EtOAc / petroleum ether) to give methyl 3-(2-fluoro-3-pyridinyl)isoxazole-4-carboxylate. LCMS [M+H] + 223.

[0372] Step 7: Synthesis of (3-(2-fluoropyridin-3-yl)isoxazol-4-yl)methanol (C29): A solution of methyl 3-(2-fluoro-3-pyridinyl)isoxazole-4-carboxylate (690 mg, 3.11 mmol) in THF (20.0 mL) was cooled to -30°C in a dry ice bath. DIBAL-H in toluene (1 M, 12.4 mL) was then added dropwise and the mixture was stirred at -30°C for 3 hours. The reaction mixture was quenched by the addition of saturated aqueous NaHCO3 solution (20 mL) from -30°C to 20°C. The mixture was then diluted with EtOAc (20 mL) and extracted with EtOAc (2×10 mL). The combined organic layers were washed with water (20 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give [3-(2-fluoro-3-pyridinyl)isoxazol-4-yl]methanol, which was used in the next step without further purification. 1 HNMR (CDCl3400MHz)δ H =8.60-8.53(m,1H),8.41-8.31(m,1H),8.16-8.10(m,1H),7.37-7.34(m,1H),4.71-4.65(m,2H). 19 F NMR (CDCl3400MHz)δ F =-67.89.

[0373] Step 8: Synthesis of 4-(bromomethyl)-3-(2-fluoropyridin-3-yl)isoxazole (C30): To a solution of [3-(2-fluoro-3-pyridinyl)isoxazol-4-yl]methanol (348 mg, 1.79 mmol) in DME (6.00 mL) was added PBr (1.21 g, 4.48 mmol) dropwise at 0°C. The mixture was then stirred at 20°C for 6 hours. The mixture was cooled to 0°C and neutralized with saturated aqueous NaHCO to pH = 7.0. DCM (20 mL) and H2O (10 mL) were added to the mixture, stirred and separated. The aqueous layer was extracted with DCM (20 mL). The combined organic layers were dried over anhydrous MgSO4, filtered and concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel (0 to 30% EtOAc / petroleum ether as eluent) to give 4-(bromomethyl)-3-(2-fluoro-3-pyridinyl)isoxazole. LCMS [M+H] + 257,259.

[0374] Step 9: Synthesis of 3-(2-fluoropyridin-3-yl)-4-methylisoxazole (P12): A mixture of 4-(bromomethyl)-3-(2-fluoro-3-pyridinyl)isoxazole (100 mg, 389 umol) and 10% wet Pd / C (50.0 mg) in MeOH (4.00 mL). The mixture was degassed and purged with H gas three times. The mixture was stirred at 20° C. under H gas (15 psi) for 5 hours. The mixture was filtered and concentrated under reduced pressure to give 3-(2-fluoro-3-pyridinyl)-4-methyl-isoxazole, which was used in the next step without further purification. LCMS [M+H] + 179.

[0375] 2-(5-Fluoro-2-(piperidin-4-yl)pyridin-3-yl)pyrazine (P13)

[0376]

[0377] Step 1: Synthesis of tert-butyl 4-(3-bromo-5-fluoro-2-pyridinyl)-3,6-dihydro-2H-pyridine-1-carboxylate (C31): To a mixture of 2,3-dibromo-5-fluoro-pyridine (500 mg, 1.96 mmol) and tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydro-2H-pyridine-1-carboxylate (606 mg, 1.96 mmol), NaCO (415 mg, 3.92 mmol) in 1,4-dioxane (10.0 mL) and H O (2.00 mL) was added Pd(dppf)Cl (226 mg, 196 umol) at 20° C. The mixture was then purged with N 3 times at 20° C. The mixture was stirred at 100° C. under N for 16 hours. The mixture was concentrated under reduced pressure.The residue was purified by flash chromatography on silica gel (0 to 30% EtOAc / petroleum ether as eluent) to give tert-butyl 4-(3-bromo-5-fluoro-2-pyridinyl)-3,6-dihydro-2H-pyridine-1-carboxylate. 1 H NMR (CDCl3400MHz)δ H =8.40(d,J=2.4Hz,1H),7.68(dd,J=2.4Hz,7.6Hz,1H),5.98(br s,1H),4.13-4.07(m,2H),3.69-3.61(m,2H),2.57-2.50(m,2H),1.49(s,9H). LCMS[M+H-56] + 304.

[0378] Step 2: Synthesis of tert-butyl 4-(5-fluoro-3-pyrazin-2-yl-2-pyridinyl)-3,6-dihydro-2H-pyridine-1-carboxylate (C32): A mixture of tert-butyl 4-(3-bromo-5-fluoro-2-pyridinyl)-3,6-dihydro-2H-pyridine-1-carboxylate (280 mg, 783 μmol), tributyl(pyrazin-2-yl)stannane (289 mg, 783 μmol), Pd(PPh3)4 (90.6 mg, 78.4 μmol), and CuI (14.9 mg, 78.4 μmol) in toluene (3.00 mL) was degassed at 20°C and purged with N2 gas three times. The mixture was then heated to 110°C and stirred at 110°C under N2 atmosphere for 16 hours. The filtrate was concentrated under reduced pressure to obtain a residue. The residue was purified by flash silica gel chromatography (0 to 50% EtOAc / petroleum ether as eluent) to give tert-butyl 4-(5-fluoro-3-pyrazin-2-yl-2-pyridinyl)-3,6-dihydro-2H-pyridine-1-carboxylate. LCMS [M+H -100] + 257.

[0379] Step 3: Synthesis of tert-butyl 4-(5-fluoro-3-(pyrazin-2-yl)pyridin-2-yl)piperidine-1-carboxylate (C33): A mixture of tert-butyl 4-(5-fluoro-3-pyrazin-2-yl-2-pyridinyl)-3,6-dihydro-2H-pyridine-1-carboxylate (124 mg, 347 umol) and wet Pd / C (50.0 mg, 10 wt%) in EtOH (5.00 mL) was degassed and purged with H gas three times, and then the mixture was stirred at 50 ° C under H gas (40 psi) for 24 hours. The mixture was filtered and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by flash silica gel chromatography (eluent: 0 to 30% EtOAc / petroleum ether) to give tert-butyl 4-(5-fluoro-3-pyrazin-2-yl-2-pyridinyl)piperidine-1-carboxylate. LCMS[M+H-100] + 259.

[0380] Step 4: Synthesis of 2-(5-fluoro-2-(piperidin-4-yl)pyridin-3-yl)pyrazine (P13): A mixture of tert-butyl 4-(5-fluoro-3-pyrazin-2-yl-2-pyridinyl)piperidine-1-carboxylate (90.0 mg, 251 umol) and HCl / 1,4-dioxane (5.00 ml, 4 M) was stirred at 20° C. under N2 for 2 hours. The mixture was concentrated under reduced pressure to give 2-[5-fluoro-2-(4-piperidinyl)-3-pyridinyl]pyrazine, which was used directly in the next step without further purification. LCMS [M+H] + 259.

[0381] Example 1: (6R)-6-[4-[5-fluoro-3-(6-methoxy-2-methyl-3-pyridinyl)-2-pyridinyl]-piperazine- Synthesis of ethyl 1-amino-2-azaspiro[3.4]octane-2-carboxylate (1)

[0382]

[0383] Step 1: Synthesis of (6R)-6-[4-[5-fluoro-3-(6-methoxy-2-methyl-3-pyridinyl)-2-pyridinyl]-piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (1): (6R)-6-[4-(3-bromo-5-fluoro-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (40.0 mg, A mixture of 1,4-dioxane (2.00 mL) and H2O (0.500 mL) was degassed and purged with N2 gas three times. The mixture was stirred at 90°C under an N2 atmosphere for 2 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent to obtain a residue. The residue was purified by preparative HPLC (column: Waters Xbridge 150×25mm10um; mobile phase: [water(NH4HCO3)-ACN]; B%: 53%-83%, 9 min) to give (6R)-6-[4-[5-fluoro-3-(6-methoxy-2-methyl-3-pyridinyl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. 1 H NMR (CDCl3400MHz)δ H =8.11(d,J=2.8Hz,1H),7.46(d,J=8.4Hz,1H),7.12(dd,J=2.8Hz,8.0Hz,1H),6.64(d,J= 8.4Hz,1H),4.08(q,J=7.2Hz,2H),3.97(s,3H),3.88-3.78(m,2H),3.74(s,2H),3.03(br s,4H),2.49(brs,1H),2.34(s,7H),2.06(br dd,J=6.8Hz,12.4Hz,1H),1.95-1.74(m,4H),1.56-1.46(m,1H),1.22(t,J=7.2Hz,3H). 19 F NMR (CDCl3400MHz)δ F =-138.654. ​​LCMS [M+H] + 484.

[0384] Example 2: 6-(4-(3-(5-cyano-2-methylpyrimidin-4-yl)pyridin-2-yl)piperazin-1-yl)-2-azaspiro [3.4] Synthesis of ethyl octane-2-carboxylate (2)

[0385]

[0386] Step 1: Synthesis of 4-(2-fluoro-3-pyridyl)-2-methyl-pyrimidine-5-carbonitrile (C34): To a mixture of 2-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (435 mg, 1.95 mmol), 4-chloro-2-methyl-pyrimidine-5-carbonitrile (200 mg, 1.30 mmol) and NaCO (414 mg, 3.91 mmol) in 1,4-dioxane (4.00 mL) and H O (0.50 mL) was added Pd(PPh) (150 mg, 130 umol) in one portion at 20°C. The mixture was degassed under vacuum and purged with N gas several times. The mixture was heated to 90°C and stirred at 90°C for 16 hours. The reaction mixture was cooled to 20°C and DCM (20 mL) was added. The mixture was washed with brine (20 mL). The organic layer was separated, dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (eluent: 0 to 10% EtOAc / petroleum ether) to give 4-(2-fluoro-3-pyridyl)-2-methyl-pyrimidine-5-carbonitrile. LCMS [M+H] + 215.

[0387] Step 2: Synthesis of 6-[4-[3-(5-cyano-2-methyl-pyrimidin-4-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (2): To a mixture of 4-(2-fluoro-3-pyridinyl)-2-methyl-pyrimidine-5-carbonitrile (130 mg, 607 umol) and 6-piperazin-1-yl-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (194 mg, 728 umol) was added DIPEA (3.00 mL) and pyridine (3.00 mL) in one portion at 20° C. under nitrogen atmosphere. The mixture was heated to 90° C. and stirred for 16 hours. The reaction mixture was concentrated in vacuo to give the crude product as a brown oil. The residue was purified by preparative HPLC (column: Phenomenex Gemini-NX 80×40 mm×3 um; mobile phase: [water (0.05% NH3H2O+10 mM NH4HCO3)-ACN]; B%: 26%-56%, 8 min) to give 6-[4-[3-(5-cyano-2-methyl-pyrimidin-4-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. 1 HNMR (CDCl3400MHz)δ H=8.91(s,1H),8.43(dd,J=2.0Hz,4.8Hz,1H),7.75(dd,J=1.6Hz,7.2Hz,1H),7.03(d d,J=4.8Hz,7.6Hz,1H),4.11(q,J=7.2Hz,2H),3.90-3.82(m,2H),3.81-3.75(m,2H), 3.22-3.13(m,4H),2.89(s,3H),2.62-2.52(m,1H),2.50-2.39(m,4H),2.14-2.06(m, 1H),1.97-1.77(m,3H),1.73-1.68(m,1H),1.57-1.48(m,1H),1.25(t,J=7.2Hz,3H). LCMS[M+H] + 462.

[0388] Example 3: (6R)-6-[4-[3-(6-methylpyrazin-2-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro [3.4] Synthesis of ethyl octane-2-carboxylate (3)

[0389]

[0390] Synthesis of (6R)-6-[4-[3-(6-methylpyrazin-2-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (3): A mixture of 2-(2-fluoro-3-pyridinyl)-6-methyl-pyrazine (50.0 mg, 264 umol), (6R)-6-piperazin-1-yl-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (70.7 mg, 264 umol), DIPEA (1.00 mL) and pyridine (1.00 mL) was heated to 130° C. and stirred at 130° C. for 16 hours. The reaction mixture was concentrated in vacuo to give the crude product as a brown oil. The residue was purified by preparative HPLC (column: Boston Prime C18 150×30 mm×5 um; mobile phase: [water (0.05% NH3H2O+10 mM NH4HCO3)-ACN]; B%: 40%-70%, 7 min) to give (6R)-6-[4-[3-(6-methylpyrazin-2-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. 1 H NMR (CDCl3400MHz)δ H=9.03(s,1H),8.36(s,1H),8.31(dd,J=2.0Hz,4.8Hz,1H),7.87(dd,J=2.0Hz,7.6Hz ,1H),7.00(dd,J=4.8Hz,7.6Hz,1H),4.10(q,J=7.2Hz,2H),3.90-3.80(m,2H),3.79 -3.74(m,2H),3.16(t,J=4.8Hz,4H),2.64(s,3H),2.60-2.38(m,5H),2.12-2.04(m, 1H), 1.96-1.78 (m, 3H), 1.73-1.65 (m, 1H), 1.57-1.46 (m, 1H), 1.24 (t, J = 7.2Hz, 3H). LCMS[M+H] + 437.

[0391] Example 4: (6R)-6-[4-[3-(1-methylpyrazol-4-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro [3.4] Synthesis of ethyl octane-2-carboxylate (4)

[0392]

[0393] Synthesis of (6R)-6-[4-[3-(1-methylpyrazol-4-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (4): At 20°C, (6R)-6-[4-(3-bromo-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (50.0 mg, 118 umol) and 1-methyl -4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazole (24.6 mg, 118 umol) in a mixture of 1,4-dioxane (2.00 mL) and H2O (400 uL) was added PdCl2(dtbpf) (7.70 mg, 11.8 umol) and K2CO3 (32.7 mg, 236 umol) at once. The mixture was then degassed under vacuum and purged with N2 gas several times. The reaction mixture was heated to 100 ° C and stirred at 100 ° C for 16 hours under a nitrogen atmosphere. The mixture was concentrated under reduced pressure at 50 ° C. The residue was purified by preparative HPLC (column: Phenomenex Gemini-NX 80×40 mm×3 um; mobile phase: [water (0.05% NH3H2O+10 mM NH4HCO3)-ACN]; B%: 26%-50%, 8 min) to give (6R)-6-[4-[3-(1-methylpyrazol-4-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. 1 H NMR (CDCl3400MHz)δ H=8.17(dd,J=1.6Hz,4.8Hz,1H),7.87(s,1H),7.75(s,1H),7.52(dd,J=1.6Hz,7 .6Hz,1H),6.89(dd,J=5.2Hz,7.6Hz,1H),4.09(q,J=7.2Hz,2H),3.98-3.94(m,3 H),3.92-3.75(m,4H),3.28-3.02(m,4H),2.74-2.31(m,5H),2.16-2.08(m,1H), 1.98-1.77(m,3H),1.76-1.66(m,1H),1.58-1.47(m,1H),1.23(t,J=7.2Hz,3H). LCMS [M+H] + 425.

[0394] Example 5: (6R)-6-[4-[3-(2-isopropyltriazol-4-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro [3.4] Synthesis of ethyl octane-2-carboxylate (5)

[0395]

[0396] Step 1: Synthesis of (R)-(2-(4-(2-(ethoxycarbonyl)-2-azaspiro[3.4]octan-6-yl)piperazin-1-yl)pyridin-3-yl)boronic acid (C35): At 25°C, to (6R)-6-[4-(3-bromo-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (100 mg, 236 μmol) and 4,4,5, To a mixture of 5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2-yl)-1,3,2-dioxaborolane (112 mg, 472 umol) in 1,4-dioxane (5.00 mL) was added Pd(PPh3)4 (27.3 mg, 23.6 umol) and KOAc (46.4 mg, 472 umol) in one portion. The mixture was then degassed under vacuum and purged with N2 gas several times. The reaction mixture was heated to 100 ° C and stirred at 100 ° C for 16 hours under a nitrogen atmosphere. The reaction mixture was cooled to 25 ° C. The reaction mixture was used directly in the next step without further purification.

[0397] Step 2: Synthesis of ethyl (6R)-6-[4-[3-(2-isopropyltriazol-4-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (5): To a mixture of [2-[4-[(6R)-2-ethoxycarbonyl-2-azaspiro[3.4]octan-6-yl]piperazin-1-yl]-3-pyridinyl]boronic acid (91.7 mg, 236 umol) in 1,4-dioxane was added 4-bromo-2-isopropyl-triazole (49.4 mg, 260 umol), H2O (500 uL), Pd(dppf)Cl2 (17.3 mg, 23.62 umol) and K2CO3 (65.3 mg, 472 umol) at 25° C. The mixture was then degassed under vacuum and purged with N2 gas several times. The reaction mixture was heated to 100°C and stirred at 100°C under a nitrogen atmosphere for 12 hours. The mixture was cooled to 25°C, DCM (5 mL) was added, and filtered. The filtrate was concentrated under reduced pressure at 50°C. The residue was purified by flash chromatography on silica gel (eluent: 0 to 10% MeOH / DCM) to give a crude product. The crude product was purified by preparative HPLC (column: Phenomenex Gemini-NX 150×30 mm×5 μm; mobile phase: [water (0.05% NH3H2O)-ACN]; B%: 45%-75%, 7 min) to give (6R)-6-[4-[3-(2-isopropyltriazol-4-yl)-2-pyridyl]piperazin-1-yl]-2-azaspiro[3.4]-octane-2-carboxylic acid ethyl ester. 1 HNMR (CDCl3400MHz)δ H =8.26(dd,J=2.0Hz,4.8Hz,1H),8.10(s,1H),7.97(dd,J=1.6Hz,7.6Hz,1H),6. 96(dd,J=4.8Hz,7.6Hz,1H),4.90-4.83(m,1H),4.10(q,J=7.2Hz,2H),3.90-3. 75(m,4H),3.19-3.12(m,4H),2.61-2.50(m,5H),2.15-2.08(m,1H),1.96-1.79 (m,3H),1.76-1.68(m,1H),1.65-1.61(m,6H),1.56-1.52(m,1H),1.24(m,3H). LCMS [M+H] + 454.

[0398] Example 6: 2-pyrazin-2-yl-6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro [3.4] Synthesis of octane (6)

[0399]

[0400] Synthesis of 2-(pyrazin-2-yl)-6-(4-(3-(pyrazin-2-yl)pyridin-2-yl)piperazin-1-yl)-2-azaspiro[3.4]octane (6): A solution of 6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane (50.0 mg, 143 umol) and 2-chloropyrazine (15.3 uL, 171 umol) in DIPEA (2.50 mL, 14.3 mmol) and pyridine (1.25 mL, 15.5 mmol) was stirred at 100° C. for 16 hours. The mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Phenomenex Gemini-NX 150×30 mm×5 um; mobile phase: [water (0.05% NH 3 H 2 O)-ACN]; B%: 23%-53%, 7 min) to give 2-pyrazin-2-yl-6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane. 1 H NMR (CDCl3400MHz)δ H =9.25(d,J=1.2Hz,1H),8.65(d,J=2.0Hz,1H),8.47(d,J=2.8Hz,1H),8.33(dd,J=2.0Hz,4.8Hz,1H),8.04 -7.89(m,1H),7.87(dd,J=2.0Hz,3.6Hz,1H),7.82(d,J=2.8Hz,1H),7.75(s,1H),7.02(dd,J=5.2Hz,7.6Hz ,1H),4.00-3.90(m,4H),3.17(t,J=4.0Hz,4H),2.63-2.57(m,1H),2.48(d,J=4.0Hz,4H),2.18(dd,J=7.2 Hz, 12.8Hz, 1H), 2.04-2.00 (m, 1H), 1.96-1.87 (m, 2H), 1.78 (dd, J = 9.6Hz, 12.8Hz, 1H), 1.57-1.50 (m, 1H). LCMS[M+H] + 429.

[0401] Example 7: 6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate Synthesis of 2,2-difluoroethyl benzoate (7)

[0402]

[0403] Synthesis of 2,2-difluoroethyl 6-(4-(3-(pyrazin-2-yl)pyridin-2-yl)piperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylate (7): A solution of 2,2-difluoroethanol (719 uL, 12.2 mmol), bis(trichloromethyl)carbonate (1.22 g, 4.11 mmol) and DMF (93.8 uL, 1.22 mmol) in DCM (6.00 mL) was cooled to 0° C. Et3N (1.70 mL, 12.2 mmol) was added and stirred at 20° C. for 3 hours. Then, a solution of 6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane (100 mg, 285 μmol) and EtN (119 μL, 856 μmol) in DCM (4.00 mL) was added. The mixture was stirred at 20° C. for 2 hours. The mixture was diluted with DCM (20 mL) and washed with saturated aqueous NaCO (20 mL), brine (20 mL), then dried over anhydrous NaSO, filtered, and concentrated in vacuo. The crude product was purified by preparative HPLC (column: Boston Prime C18 150×30 mm×5 um; mobile phase: [water (0.05% NH3H2O+10 mM NH4HCO3)-CAN]; B%: 35%-65%, 7 min) to give 2,2-difluoroethyl 6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate. 1 H NMR (CDCl3400MHz)δ H =9.23(s,1H),8.67(dd,J=2.8Hz,1H),8.46(d,J=2.8Hz,1H),8.33(dd,J=2.0Hz,5.2H z,1H),7.87(dd,J=1.6Hz,7.6Hz,1H),7.02(dd,J=4.8Hz,7.2Hz,1H),6.06-5.77(m,1H ),4.26-4.18(m,2H),3.92-3.81(m,4H),3.16(t,J=4.8Hz,4H),2.55-2.45(m,5H),2. 08(dd,J=4.8Hz,7.2Hz,1H),1.95-1.79(m,3H),1.72-1.67(m,1H),1.53-1.49(m,1H). 19 F NMR (CDCl3400MHz)δ F =-126.01. LCMS [M+H] + 459.

[0404] Examples 8, 9 and 10: 6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane Alkane-2-carboxylic acid but-2-ynyl ester (8), (6R)-6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro [3.4] octane-2-carboxylic acid but-2-ynyl ester (9) and (6S)-6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2- Synthesis of azaspiro[3.4]octane-2-carboxylic acid but-2-ynyl ester (10)

[0405]

[0406] Step 1: Synthesis of but-2-ynyl 6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (8): A solution of but-2-yn-1-ol (21.0 mg, 300 umol) and CDI (48.6 mg, 300 umol) in DCM (2.00 mL) was stirred at 20° C. for 1 hour. EtN (36.4 mg, 360 umol) and 6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane (105 mg, 300 umol) in DCM (1.00 mL) were then added. The mixture was then stirred at 20° C. for 16 hours. The mixture was diluted with DCM (40 mL), washed with saturated aqueous NaHCO₃ (20 mL), dried over Na₂SO₄, filtered, and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by preparative HPLC (column: Phenomenex C18 75×30 mm×3 μm; mobile phase: [water (NH₃H₂O+NH₄HCO₃)-ACN]; B%: 37%-67%, 7 min) to give but-2-ynyl 6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate. 1 H NMR (CDCl3400MHz)δ H =9.24(d,J=1.2Hz,1H),8.65(dd,J=1.6Hz,2.4Hz,1H),8.47(d,J=2.4Hz,1H),8.33(dd,J=1.6H z,4.8Hz,1H),7.87(dd,J=1.6Hz,7.6Hz,1H),7.01(dd,J=5.2Hz,7.6Hz,1H),4.65-4.60(m,2H) ,3.93-3.82(m,2H),3.80(s,2H),3.15(t,J=4.8Hz,4H),2.61-2.50(m,1H),2.50-2.40(m,3H), 2.49-2.40(m,1H),2.12-2.04(m,1H),1.95-1.78(m,6H),1.71-1.66(m,1H),1.54-1.45(m,1H). LCMS[M+H] + 447.

[0407] Step 2: Synthesis of (6R)-6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]-octane-2-carboxylic acid but-2-ynyl ester (9) and (6S)-6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid but-2-ynyl ester (10): But-2-ynyl 6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (500 mg, 1.12 mmol) was purified by SFC (column: Phenomenex-cellulose-2 (250 mm×30 mm, 10 μm); mobile phase: [0.1% NH 3 H 2 O EtOH]; B%: 40%-40%, min) to obtain Peak 1 (6R)-6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid but-2-ynyl ester and Peak 2 (6S)-6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid but-2-ynyl ester. Peak 1: 1 HNMR (CDCl3400MHz)δ H =9.23(brs,1H),8.65(brs,1H),8.48(br s,1H),8.33(br d,J=5.2Hz,1H),7.87(d,J=7.6Hz,1H),7.05-6.97(m,1H),4.62(br s,2H),3.94-3.73(m,4H),3.16(br s,4H),2.65-2.35(m 5H),2.07(br s,1H),1.96-1.75(m,6H),1.67(brs,1H),1.53-1.44(m,1H). LCMS[M+H] + 447. Peak 2: 1 H NMR (CDCl3400MHz)δ H =9.23(brs,1H),8.66(brs,1H),8.48(br s,1H),8.33(dd,J=1.6Hz,4.8Hz,1H),7.87(dd,J=2.0Hz,7.6Hz,1H),7.07-6 .97(m,1H),4.62(d,J=2.8Hz,2H),3.94-3.75(m,4H),3.16(brs,4H),2.46(br s,5H),2.08(brs,1H),1.99-1.74(m,6H),1.68(brs,1H),1.53-1.44(m,1H). LCMS[M+H] +447.

[0408] Examples 11 and 12: (6R)-6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro [3.4]-octane (11) and oxetane-3-yl-[(6R)-6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]- Synthesis of 2-azaspiro[3.4]octan-2-yl]methanone (12)

[0409]

[0410] Step 1: Synthesis of (6R)-6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane (11): To a solution of tert-butyl (6R)-6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (200 mg, 444 umol) in DCM (4.00 mL) was added TFA (1.00 mL, 13.5 mmol) and the mixture was stirred at 20° C. for 2 hours. The mixture was adjusted to pH=9 with 10% aqueous NaOH. The mixture was extracted with DCM (2×50 mL). The combined organic layers were dried over anhydrous Na 2 SO 4 and filtered. The filtrate was concentrated under reduced pressure to obtain (6R)-6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane. 1 H NMR (CDCl3400MHz)δ H =9.24(d,J=1.2Hz,1H),8.65(s,1H),8.47(d,J=2.4Hz,1H),8.32(dd,J=1.6Hz,4.8Hz,1H),7.86(dd,J=1.6Hz,7.2Hz,1H),7.00(dd,J=4.8H z,7.2Hz,1H),3.74-3.30(m,4H),3.15(d,J=4.4Hz,4H),2.46(s,5H),2.07-1.69(m,4H),1.61(dd,J=10.0,12.4Hz,1H),1.50-1.36(m,1H). LCMS[M+H] + 351.

[0411] Step 2: Synthesis of oxetane-3-yl-[(6R)-6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octan-2-yl]methanone (12): A mixture of oxetane-3-carboxylic acid (8.74 mg, 85.6 umol), EDCI (24.62 mg, 128.40 umol), HOBt (3.47 mg, 25.7 umol) and DIPEA (45.0 uL, 257 umol) in DMF (1.00 mL) was stirred at 20 °C for 0.5 hours. Then, (6R)-6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane (30.0 mg, 85.6 μmol) was added to the mixture, and the mixture was stirred at 20° C. for 16 hours. The mixture was purified by preparative HPLC (column: Welch Xtimate C18 150×30 mm×5 μm; mobile phase: [water(NH 3 H 2 O+NH 4 HCO 3 )-ACN]; B%: 13%-43%, 9 min) to give oxetan-3-yl-[(6R)-6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octan-2-yl]methanone. 1 HNMR (CDCl3400MHz)δ H =9.24(s,1H),8.66(s,1H),8.48(br s,1H),8.33(d,J=3.2Hz,1H),7.88(d,J=7.2Hz,1H),7.03(d,J=4.0Hz,1H), 4.94-4.84(m,2H),4.72(dd,J=6.4,8.0Hz,2H),3.95-3.73(m,5H),3.16(br s,4H),2.67-2.32(m,5H),2.17-1.99(m,1H),1.97-1.78(m,3H),1.75-1.66(m,1H),1.54-1.43(m,1H). LCMS[M+H] + 435.

[0412] Example 13: Ethyl 2-[(6R)-6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro Synthesis of [3.4]octan-2-yl]oxazole (13)

[0413]

[0414] Synthesis of ethyl 2-[(6R)-6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octan-2-yl]oxazole (13): To a solution of (6R)-6-[4-(3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane (96.0 mg, 274 umol) and 2-bromooxazole (48.6 mg, 329 umol) in 1,4-dioxane (2.00 mL) was added t-BuONa (79.0 mg, 822 umol), XantPhos (19.0 mg, 32.8 umol) and Pd2(dba)3 (25.1 mg, 27.4 umol) at 20° C. The mixture was degassed under vacuum and purged with N2 gas several times. The mixture was then stirred at 90°C for 16 hours. The mixture was cooled to 20°C. The reaction mixture was filtered and the filter cake was washed with THF (15 mL). The filtrate was then concentrated under reduced pressure to obtain a crude product. The crude product was purified by preparative HPLC (column: Welch Xtimate C18 150×30 mm×5 μm; mobile phase: [water (NH3H2O+NH4HCO3)-ACN]; B%: 25%-55%, 25 min) to obtain 2-[(6R)-6-[4-(3-pyrazin-2-yl-2-pyridinyl)-piperazin-1-yl]-2-azaspiro[3.4]octan-2-yl]oxazole. 1 H NMR (CDCl3400MHz)δ H =9.24(s,1H),8.67-8.62(m,1H),8.47(d,J=2.8Hz,1H),8.33(dd,J=1.6Hz,4.8Hz,1H),7 .87(dd,J=1.6Hz,7.6Hz,1H),7.19(s,1H),7.01(dd,J=4.8Hz,7.6Hz,1H),6.78(s,1H),4 .01-3.93(m,2H),3.93-3.88(m,2H),3.17(s,4H),2.60-2.43(m,5H),2.16(dd,J=6.8Hz, 12.8Hz,1H),2.02-1.95(m,1H),1.92-1.84(m,2H),1.74(d,J=10.8Hz,1H),1.52(s,1H). LCMS[M+H] + 418.

[0415] Example 14: (6R)-6-[4-[3-(4-ethoxycarbonylpyrazol-1-yl)-2-pyridinyl]piperazin-1-yl]-2- Synthesis of Azaspiro[3.4]-octane-2-carboxylate (14)

[0416]

[0417] Synthesis of (6R)-6-[4-[3-(4-ethoxycarbonylpyrazol-1-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (14): (6R)-6-[4-(3-bromo-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (50.0 mg, 118 μmol) was added to the mixture at 20°C. To a mixture of (1R, 2R)-N1, N2-dimethylcyclohexane-1,2-diamine (3.36 mg, 23.6 umol) in DMF (1.00 mL) was added CuI (2.25 mg, 11.8 umol) and K3PO4 (75.2 mg, 354 umol) at one time. The mixture was then degassed under vacuum and purged with N2 gas several times. The reaction mixture was heated to 130 ° C and stirred at 130 ° C for 16 hours under a nitrogen atmosphere. The mixture was cooled to 25 ° C, DCM (3.00 mL) was added and filtered. The filtrate was concentrated under reduced pressure at 50 ° C. The residue was purified by preparative HPLC (column: Phenomenex Gemini-NX 80×40 mm×3 um; mobile phase: [water (0.05% NH3H2O)-ACN]; B%: 32%-62%, 8 min) to give (6R)-6-[4-[3-(4-ethoxycarbonylpyrazol-1-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. 1 HNMR (CDCl3400MHz)δ H =8.50(s,1H),8.29(dd,J=1.6Hz,4.8Hz,1H),8.10(s,1H),7.72(dd,J=1.6Hz,7.6Hz,1H),6. 97(dd,J=4.8Hz,7.6Hz,1H),4.35(q,J=7.2Hz,2H),4.09(q,J=7.2Hz,2H),3.88-3.72(m,4H), 3.05-2.94(m,4H),2.58-2.39(m,5H),2.09(dd,J=6.8Hz,12.8Hz,1H),1.94-1.77(m,3H),1.6 8(dd,J=9.6Hz,12.8Hz,1H),1.52-1.47(m,1H),1.38(t,J=7.2Hz,3H),1.23(t,J=7.2Hz,3H). LCMS[M+H] + 483.

[0418] Example 15: 6-[4-[3-(cyclopropyloxy)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-yl Synthesis of ethyl formate (15)

[0419]

[0420] Synthesis of ethyl 6-[4-[3-(cyclopropyloxy)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (15): A mixture of ethyl 6-[4-(3-hydroxy-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (80.0 mg, 222 umol), bromocyclopropane (269 mg, 2.22 mmol, 178 uL) and CsCO (217 mg, 666 umol) in DMA (3.00 mL) was heated to 130° C. and stirred at 130° C. for 14 hours. The mixture was cooled to 20° C. and DCM (20.0 mL) was added to the mixture, filtered and concentrated in vacuo to give the crude product (100 mg). The crude product was purified by preparative HPLC (column: Phenomenex Gemini-NX 80×40 mm×3 μm; mobile phase: [water (0.05% NH 3 H 2 O + 10 mM NH 4 HCO 3 )-ACN]; B%: 35%-61%, 8 min). The fractions containing the product were then combined, concentrated in vacuo, and freeze-dried to give ethyl 6-[4-[3-(cyclopropyloxy)-2-pyridyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate. 1 H NMR (CDCl3400MHz)δ H =7.88(dd,J=1.2Hz,5.2Hz,1H),7.41(d,J=8.0Hz,1H),6.83(dd,J=4.8Hz,7.6Hz,1H) ,4.10(q,J=7.2Hz,2H),3.95-3.87(m,1H),3.86-3.82(m,1H),3.81-3.75(m,2H),3.74 -3.68(m,1H),3.60-3.16(m,4H),2.81-2.41(m,5H),2.20-2.09(m,1H),2.02-1.88(m ,2H),1.87-1.65(m,2H),1.62-1.58(m,1H),1.24(t,J=7.2Hz,3H),0.85-0.75(m,4H). LCMS[M+H] + 401.

[0421] Examples 16, 17 and 18: 6-[4-[5-fluoro-3-(1,3,4-thiadiazol-2-yl)-2-pyridinyl]piperazin-1-yl]- 2-Azaspiro[3.4]octane-2-carboxylic acid ethyl ester (16), (6S)-6-[4-[5-fluoro-3-(1,3,4-thiadiazol-2-yl)-2-pyridine 1-yl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (17) and (6R)-6-[4-[5-fluoro-3-(1,3,4-thiadiol)] Synthesis of ethyl [oxazol-2-yl]-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (18)

[0422]

[0423] Step 1: Synthesis of ethyl 6-[4-[5-fluoro-3-(1,3,4-thiadiazol-2-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (16): A mixture of 2-(5-fluoro-2-piperazin-1-yl-3-pyridinyl)-1,3,4-thiadiazol (60.0 mg, 226 μmol) and ethyl 6-oxo-2-azaspiro[3.4]octane-2-carboxylate (37.2 mg, 188 μmol) in DCE (2.00 mL) was stirred at 20° C. for 1 hour. AcOH (5.66 mg, 94.2 μmol) and NaBH(OAc) 3 (120 mg, 565 μmol) were then added to the reaction mixture. The mixture was stirred at 20° C. for 16 hours. The mixture was poured into saturated aqueous NaHCO₃ (30 mL) and stirred for 5 min. The mixture was extracted with DCM (3 × 30 mL). The combined organic phases were washed with brine (100 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated in vacuo to yield a residue. The residue was purified by flash chromatography on silica gel (eluent: 0 to 3% MeOH / DCM) to yield the crude product. The crude product was purified by preparative HPLC (column: Phenomenex C18 80 × 40 mm × 3 μm; mobile phase: [water (NH₃H₂O)-ACN]; B%: 36%-66% over 8 min) to yield ethyl 6-[4-[5-fluoro-3-(1,3,4-thiadiazol-2-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate. 1 HNMR (CDCl3400MHz)δ H =9.21(s,1H),8.33(brd,J=2.0Hz,2H),4.10(q,J=7.2Hz,2H),3.97-3.82(m,2H),3. 79(s,2H),3.11(brs,4H),2.66(brs,5H),2.15(brs,1H),2.03-1.83(m,3H),1.76(br s,1H),1.64-1.58(m,1H),1.24(t,J=6.8Hz,3H). 19 F NMR (CDCl3400MHz)δ F =-132.678. LCMS [M+H] + 447.

[0424] Step 2: Synthesis of (6S)-6-[4-[5-fluoro-3-(1,3,4-thiadiazol-2-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (17) and (6R)-6-[4-[5-fluoro-3-(1,3,4-thiadiazol-2-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (18): 6-[4-[5-fluoro-3-(1,3,4-thiadiazol-2-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (150 mg, 336 umol) was purified by SFC (column: DAICEL CHIRALCEL OD-H (250mm×30mm, 5um); mobile phase: [0.1% NH3H2O ​​EtOH]; B%: 30%-30%, min) purification was performed to obtain Peak 1 (6S)-6-[4-[5-fluoro-3-(1,3,4-thiadiazol-2-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester and Peak 2 (6R)-6-[4-[5-fluoro-3-(1,3,4-thiadiazol-2-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. Peak 1: 1 H NMR (CDCl3400MHz)δ H =9.21(s,1H),8.33(d,J=2.0Hz,2H),4.10(q,J=7.2Hz,2H),3.93-3.75(m,4H),3.10(s,4H),2.65(s,5H),2.13(br s,1H),2.01-1.78(m,3H),1.74(s,2H),1.24(t,J=7.2Hz,3H). 19 FNMR (CDCl3400MHz)δ F =-132.66. LCMS [M+H] + 447. Peak 2: 1 HNMR (CDCl3400MHz)δ H =9.21(s,1H),8.33(d,J=2.0Hz,2H),4.10(q,J=7.2Hz,2H),3.93-3.75(m,4H),3.10(s,4H),2.65(s,5H),2.13(br s,1H),2.01-1.78(m,3H),1.74(s,2H),1.24(t,J=7.2Hz,3H). 19 F NMR (CDCl3400MHz)δ F =-132.66. LCMS [M+H] + 447.

[0425] Example 19: 6-(4-(5'-fluoro-[3,3'-bipyridyl]-2-yl)piperazin-1-yl)-2-azaspiro[3.4]octane- Synthesis of ethyl 2-formate (19)

[0426]

[0427] Synthesis of ethyl 6-(4-(5'-fluoro-[3,3'-bipyridyl]-2-yl)piperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylate (19): To a mixture of ethyl 6-[4-(3-bromo-2-pyridyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (50.0 mg, 118 umol), (5-fluoro-3-pyridyl)boronic acid (33.3 mg, 236 umol) and Na2CO3 (37.6 mg, 354 umol) in 1,4-dioxane (4.00 mL) and H2O (1.00 mL) was added [1,1'-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) (7.70 mg, 11.8 umol) in one portion under nitrogen atmosphere at 20°C. The mixture was then degassed under vacuum and purged with N2 gas several times. The mixture was stirred at 80°C for 16 hours. The reaction mixture was concentrated. The crude product was purified by preparative HPLC (column: Phenomenex Gemini-NX 80×40 mm×μm. Conditions: water (0.05% NH3H2O ​​+ 10 mM NH4HCO3)-ACN. Start B: 26; End B: 56. Gradient time (min): 8. 100% B hold time (min): 3. Flow rate (ml / min): 30) to give ethyl 6-[4-[3-(5-fluoro-3-pyridinyl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate. 1 H NMR (CDCl3400MHz)δ H=8.66(s,1H),8.45(d,J=2.4Hz,1H),8.31(dd,J=2.0Hz,4.8Hz,1H),7.73(brd,J=9.6Hz,1H),7.49(dd,J=1.6Hz,7.2Hz,1H),6.99(dd,J=4.8Hz,7.2Hz,1H),4.10(q,J=7.2Hz,2H),3.92-3.86(m,1H),3.86-3.80(m,1H),3.80-3.74(m,2H),3.20-3.10(m,4H),2.65-2.34(m,5H),2.10(br dd,J=7.2Hz,12.4Hz,1H),1.97-1.78(m,3H),1.74-1.65(m,1H),1.55-1.45(m,1H),1.24(t,J=7.2Hz,3H)。LCMS[M+H] + 440。

[0428] Example 20: 6-[4-[3-(cyclobutylamino)-2-pyridyl]piperazin-1-yl]-2-azaspiro-[3.4]octane- Synthesis of ethyl 2-formate (20)

[0429]

[0430] Synthesis of ethyl 6-[4-[3-(cyclobutylamino)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (20): A mixture of ethyl 6-[4-(3-bromo-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (50.0 mg, 118 umol), cyclobutylamine (42.0 mg, 590 umol), t-BuONa (34.1 mg, 354 umol), 2-[bis(3,5-trifluoromethylphenylphosphino)-3,6-dimethoxy]-2,6-dimethylamino-1,1-biphenyl (L7, 8.93 mg, 11.8 umol) and Pd2(dba)3 (10.8 mg, 11.8 umol) in 1,4-dioxane (3.00 mL). The mixture was degassed under vacuum and purged with N2 gas several times. The mixture was then heated to 100°C and stirred at 100°C for 16 hours. The mixture was cooled to 20°C, and DCM (5 mL) was added to the mixture, filtered and concentrated in vacuo to give the crude product. The crude product was purified by preparative HPLC (column: Phenomenex Gemini-NX 80×40 mm×3 μm; mobile phase: [water (0.05% NH3H2O+10 mM NH4HCO3)-ACN]; B%: 48%-72%, 8 min). The fractions containing the product were then combined, concentrated in vacuo and freeze-dried to give ethyl 6-[4-[3-(cyclobutylamino)-2-pyridyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate. 1 HNMR (CDCl3400MHz)δ H =7.70(d,J=0.4Hz,1H),6.86(dd,J=4.8Hz,7.6Hz,1H),6.69(d,J=8.0Hz,1H),4.33 -4.21(m,1H),4.10(q,J=7.2Hz,2H),3.95-3.89(m,1H),3.89-3.82(m,2H),3.82-3. 76(m,2H),3.33-3.29(m,4H),3.28-2.50(m,4H),2.50-2.35(m,2H),2.21-2.09(m,1 H), 1.99-1.90 (m, 2H), 1.90-1.72 (m, 6H), 1.68-1.60 (m, 2H), 1.24 (t, J = 6.8Hz, 3H). LCMS[M+H] + 414.

[0431] Example 21: 6-[4-[3-(2,6-dimethylphenyl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro-[3.4] Synthesis of ethyl octane-2-carboxylate (21)

[0432]

[0433] Synthesis of ethyl 6-[4-[3-(2,6-dimethylphenyl)-2-pyridyl]piperazin-1-yl]-2-azaspiro-[3.4]octane-2-carboxylate (21): A mixture of 6-[4-[3-(2,6-dimethylphenyl)-2-pyridyl]piperazin-1-yl]-2-azaspiro[3.4]octane (75.0 mg, 199 umol, TFA salt), TEA (60.5 mg, 598 umol) in DCM (5.00 mL) was added at 0-5°C. Ethyl chloroformate (340 mg, 3.13 mmol) was then added dropwise to the mixture at 0-5°C. The mixture was then warmed to 20°C and stirred at 20°C for 5 hours. The reaction was slowly quenched with aqueous NaHCO3 (10%, 25 mL) and then extracted with DCM (50 mL). The organic phase was washed with brine (20 mL), dried over anhydrous Na2SO4, filtered and concentrated in vacuo. The residue was purified by preparative HPLC (column: Phenomenex Gemini-NX 80×40 mm×3 μm; mobile phase: [water (0.05% NH3H2O+10 mM NH4HCO3)-ACN]; B%: 53%-77%, 8 min) to give ethyl 6-[4-[3-(2,6-dimethylphenyl)-2-pyridyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate. 1 H NMR (CDCl3400MHz)δ H =8.26(dd,J=1.6Hz,4.8Hz,1H),7.23(dd,J=1.6Hz,7.2Hz,1H),7.19-7.14(m,1H),7.13-7. 08(m,2H),6.87(dd,J=4.8Hz,7.2Hz,1H),4.08(q,J=3.2Hz,2H),3.87-3.77(m,2H),3.76-3 .69(m,2H),3.18-3.06(m,4H),2.54-2.33(m,1H),2.31-2.16(m,4H),2.08(s,6H),2.04-1. 99(m,1H),1.91-1.75(m,3H),1.69-1.61(m,1H),1.53-1.42(m,1H),1.22(t,J=7.2Hz,3H). LCMS[M+H] + 449.

[0434] Example 22: (6R)-6-[4-(6-Fluoro-3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro [3.4] Synthesis of ethyl octane-2-carboxylate (22)

[0435]

[0436] Step 1: Synthesis of (6R)-6-[4-(3-bromo-6-fluoro-2-pyridyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (C36): Under a nitrogen atmosphere, to a solution of 3-bromo-2,6-difluoro-pyridine (100 mg, 516 umol) in DMSO (3.00 mL) and K2CO3 (214 mg, 1.55 mmol) at 20°C was added (6R)-6-piperazin-1-yl-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (152 mg, 567 umol) in one portion. The mixture was degassed and purged with N2 gas several times. The mixture was stirred at 20°C for 16 hours. The reaction mixture was poured into water (100 mL) at 20°C and then extracted with EtOAc (3×100 mL). The combined organic layers were washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by flash chromatography on silica gel (eluent: 0 to 100% EtOAc / petroleum ether) to give (6R)-6-[4-(3-bromo-6-fluoro-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. LCMS [M+H] + 441,443.

[0437] Step 2: Synthesis of (6R)-6-[4-(6-fluoro-3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (22): Under nitrogen atmosphere, to a solution of (6R)-6-[4-(3-bromo-6-fluoro-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]-octane-2-carboxylic acid ethyl ester (100 mg, 227 umol) in toluene (3.00 mL), Pd(PPh3)4 (26.2 mg, 22.7 umol) and CuI (14.9 mg, 78.4 umol) in toluene (3.00 mL) was added tributyl(pyrazin-2-yl)stannane (100 mg, 272 umol) in one portion at 20°C. The mixture was heated to 110° C. and stirred at 110° C. for 12 hours. The reaction mixture was poured onto water (100 mL) at 20° C. and then extracted with EtOAc (3×100 mL). The combined organic layers were washed with brine (100 mL), dried over Na 2 SO 4 , filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Welch Xtimate C18 150×25 mm×5 μm; mobile phase: [water (NH 3 H 2 O + NH 4 HCO 3 )-ACN]; B%: 45%-75%, 7 min) to give (6R)-6-[4-(6-fluoro-3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. 1HNMR (CDCl3400MHz)δ H =9.11(d,J=1.2Hz,1H),8.76-8.61(m,1H),8.46(d,J=2.4Hz,1H),7.94(t,J =8.0Hz,1H),6.56(dd,J=3.2Hz,8.0Hz,1H),4.08(q,J=7.2Hz,2H),3.90-3.6 9(m,4H),3.30-3.05(m,4H),2.73-2.32(m,5H),2.07(dd,J=7.2,12.4Hz,1H ), 1.95-1.76 (m, 3H), 1.67-1.61 (m, 1H), 1.51 (s, 1H), 1.22 (t, J = 7.2Hz, 3H). 19 FNMR (CDCl3400MHz)δ F =-67.666. LCMS [M+H] + 441.

[0438] Example 23: (6R)-6-[4-(3-isoxazol-5-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro-[3.4] Synthesis of ethyl octane-2-carboxylate (23)

[0439]

[0440] Synthesis of ethyl (6R)-6-[4-(3-isoxazol-5-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro-[3.4]octane-2-carboxylate (23): A mixture of ethyl (6R)-6-[4-[3-[(E)-3-(dimethylamino)prop-2-enoyl]-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (100 mg, 226 μmol) in anhydrous MeOH (2.00 mL) was cooled to 0-5° C. in an ice-water bath, and then a solution of aminohydrogensulfate (28.2 mg, 249 μmol) in anhydrous MeOH (1.00 mL) was added dropwise at 0-5° C. The mixture was then warmed to 20° C. and stirred at 20° C. for 20 hours. The reaction mixture was concentrated under reduced pressure. To the residue was added saturated aqueous NH4Cl solution (10 mL) and extracted with DCM (2×10 mL). The combined organic layers were washed with brine (10 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to give a residue as a brown oil. The residue was purified by preparative HPLC (column: Xtimate C18 100×30 mm×3 μm; mobile phase: [water (0.225% FA)-ACN]; B%: 5%-35%, 8 min) to give (6R)-6-[4-(3-isoxazol-5-yl-2-pyridyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. 1H NMR (CDCl3400MHz)δ H =8.41-8.30(m,2H),8.02(dd,J=2.0Hz,8.0Hz,1H),7.01(dd,J=4.8Hz,7.6Hz,1H),6.81(s,1H),4.12(q,J=7.2Hz,2H),3.97-3.73(m,4H),3.24(br s,4H),2.59(br s,5H),2.16(br dd,J=6.8Hz,12.4Hz,1H),2.03-1.82(m,3H),1.80-1.70(br s,1H),1.57-1.49(m,1H),1.26(t,J=7.2Hz,3H). LCMS[M+H] + 412.

[0441] Example 24: (6R)-6-[4-[3-[1-(2-methoxyethyl)pyrazol-4-yl]-2-pyridinyl]piperazine-1-yl Synthesis of ethyl]-2-azaspiro[3.4]octane-2-carboxylate (24)

[0442]

[0443] Step 1: Synthesis of (6R)-6-[4-[3-(1H-pyrazol-4-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (C37): Under nitrogen atmosphere, to (6R)-6-[4-(3-bromo-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (300 mg, 709 μmol), 4 To a mixture of tert-butyl-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazole-1-carboxylate (208 mg, 709 umol) and K2CO3 (196 mg, 1.42 mmol) in 1,4-dioxane (5.00 mL) and water (1.00 mL) was added Pd(dppf)Cl2 (51.9 mg, 70.9 umol) in one portion. The mixture was degassed under vacuum and purged with N2 gas several times. The mixture was then stirred at 100°C for 4 hours. The reaction mixture was filtered and the filter cake was washed with DCM (3×15 mL). The filtrate was then concentrated under reduced pressure to obtain the crude product. The crude product was purified by flash chromatography on silica gel (0 to 5% MeOH / DCM as eluent) to give (6R)-ethyl 6-[4-[3-(1H-pyrazol-4-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate. LCMS [M+H] + 411.

[0444] Step 2: Synthesis of (6R)-6-[4-[3-[1-(2-methoxyethyl)pyrazol-4-yl]-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (24): A mixture of (6R)-6-[4-[3-(1H-pyrazol-4-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]-octane-2-carboxylic acid ethyl ester (40 mg, 97.4 μmol), 1-bromo-2-methoxy-ethane (16.2 mg, 117 μmol), and CsCO (63.5 mg, 195 μmol) in DMF (2.00 mL) was stirred at 85° C. under N atmosphere for 12 hours. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give the crude product. The crude product was purified by preparative HPLC (column: Welch Xtimate C18 150×30 mm×5 um; mobile phase: [water (NH3H2O+NH4HCO3)-ACN]; B%: 27%-57%, 7 min) to give (6R)-6-[4-[3-[1-(2-methoxyethyl)pyrazol-4-yl]-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. 1 HNMR (CDCl3400MHz)δ H =8.16(dd,J=1.6Hz,4.8Hz,1H),7.91(s,1H),7.87(s,1H),7.55(dd,J=2.0,7.6Hz,1H ),6.90(dd,J=4.8Hz,7.2Hz,1H),4.32(t,J=5.2Hz,2H),4.09(q,J=7.2Hz,2H),3.91-3 .81(m,2H),3.80-3.75(m,4H),3.36(s,3H),3.17(s,4H),2.54(s,5H),2.11(dd,J=7. 2Hz, 12.8Hz, 1H), 1.96-1.81 (m, 3H), 1.73 (s, 1H), 1.67 (s, 1H), 1.23 (t, J = 7.2Hz, 3H). LCMS[M+H] + 469.

[0445] Examples 25 and 26: (6R)-6-[2,2,3,3,5,5,6,6-octadeuterated-4-(5-hydroxy-3-pyrazin-2-yl-2- pyridyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (25) and (6S)-6-[2,2,3,3,5,5,6,6- Octadeuterated 4-(5-hydroxy-3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester Synthesis of (26)

[0446]

[0447]

[0145] Step 1: Synthesis of ethyl 6-[2,2,3,3,5,5,6,6-octadeutero-4-(5-hydroxy-3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (C38): To a solution of ethyl 6-[4-(5-chloro-3-pyrazin-2-yl-2-pyridinyl)-2,2,3,3,5,5,6,6-octadeutero-piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (400 mg, 860 umol) in 1,4-dioxane (4.00 mL) was added a solution of KOH (145 mg, 2.58 mmol) in water (4.00 mL) in one portion under nitrogen atmosphere at 20 °C. Then di-tert-butyl-[2,3,4,5-tetramethyl-6-(2,4,6-triisopropylphenyl)phenyl]-phosphine (Me4-t-BuXphos, 82.7 mg, 172 umol) and tris(dibenzylideneacetone)dipalladium (39.4 mg, 43.0 umol) were added at a time at 20 ° C. The mixture was degassed at 25 ° C and purged with N2 gas several times. The mixture was heated to 100 ° C and stirred at 100 ° C for 12 hours. The reaction mixture was poured onto water (20 mL) at 20 ° C and then extracted with EtOAc (3 × 15 mL). The combined organic layer was washed with brine (30 mL), dried over Na2SO4, filtered and concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel (eluent: 0 to 20% MeOH in DCM) to give ethyl 6-[2,2,3,3,5,5,6,6-octadeutero-4-(5-hydroxy-3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate. LCMS [M+H] + 447.

[0448] Step 2: (6R)-6-[2,2,3,3,5,5,6,6-octadeutero-4-(5-hydroxy-3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (25) and (6S)-6-[2,2,3,3,5,5,6,6-octadeutero-4-(5-hydroxy-3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate Synthesis of 6-[2,2,3,3,5,5,6,6-octadeuterated-4-(5-hydroxy-3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (26): Ethyl 6-[2,2,3,3,5,5,6,6-octadeuterated-4-(5-hydroxy-3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate was purified by SFC (column: DAICEL CHIRALCEL OD-H (250 mm × 30 mm, 5 μm); mobile phase: [0.1% NH 3 H 2 O MeOH]; B%: 30%-30%, min) to give (6R)-6-[2,2,3,3,5,5,6,6-octadeutero-4-(5-hydroxy-3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (13.3 mg, 28.2 umol, 25.2% yield, 94.7% purity) and (6S)-6-[2,2,3,3,5,5,6,6-octadeutero-4-(5-hydroxy-3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester as a yellow solid. Peak 1: 1 H NMR (CDCl3400MHz)δ H =9.40-9.30(m,1H),8.72-8.56(m,1H),8.48(d,J=2.6Hz,1H),7.98(d,J=2.8Hz,1H),7.65-7.41(m,1H),4.09(q,J=7.2Hz,2 H), 3.93-3.59 (m, 4H), 2.60 (s, 1H), 2.09 (dd, J = 6.8Hz, 12.4Hz, 1H), 1.98-1.66 (m, 4H), 1.56 (s, 1H), 1.23 (t, J = 7.2Hz, 3H). LCMS[M+H] + 447. Peak 2: 1 H NMR (CDCl3400MHz)δ H=9.40-9.30(m,1H),8.72-8.56(m,1H),8.48(d,J=2.4Hz,1H),7.98(d,J=2.8Hz,1H),7.65-7.41(m,1H),4.09(q,J=7.2Hz,2 H), 3.93-3.59 (m, 4H), 2.60 (s, 1H), 2.09 (dd, J = 6.8Hz, 12.4Hz, 1H), 1.98-1.66 (m, 4H), 1.56 (s, 1H), 1.23 (t, J = 7.2Hz, 3H). LCMS[M+H] + 447.

[0449] Example 27: (6R)-6-(4-(3-(1-hydroxyethyl)pyridin-2-yl)piperazin-1-yl)-2-azaspiro[3.4]octane Synthesis of ethyl 2-oxanecarboxylate (27)

[0450]

[0451] Synthesis of ethyl (6R)-6-(4-(3-(1-hydroxyethyl)pyridin-2-yl)piperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylate (27): To a mixture of ethyl (6R)-6-[4-(3-acetyl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (100 mg, 259 umol) in MeOH (5.00 mL) was added NaBH4 (11.7 mg, 310 umol) and stirred at 25°C for 16 hours. More NaBH4 (11.7 mg, 310 umol) was added to the mixture and heated to 40°C, and then the reaction mixture was stirred at 40°C for 1 hour. Water (20 mL) was added to the mixture and extracted with EtOAc (2×20 mL). The combined organic phases were washed with brine (2×20 mL), dried over anhydrous Na 2 SO 4 , filtered, and concentrated under reduced pressure to give a crude product. The crude product was purified by preparative HPLC (column: Phenomenex Gemini-NX 80×30 mm×3 μm, mobile phase: [water (10 mM NH 4 HCO 3 )-ACN]; B%: 30%-60%, 9 min) to give (6R)-6-[4-[3-(1-hydroxyethyl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. 1 HNMR (CDCl3400MHz)δ H=8.29(dd,J=1.6Hz,4.8Hz,1H),7.61-7.56(m,1H),7.06(dd,J=4.8Hz,7.2Hz,1H),5.07(q,J=6.8Hz,1H),4.10(q,J=7.2Hz,2H),3.93-3.82(m, 2H),3.82-3.76(m,2H),3.30-3.04(m,4H),2.67(brs,5H),2.17-2.09( m, 1H), 1.95-1.70 (m, 5H), 1.54 (d, J = 4.8Hz, 3H), 1.24 (t, J = 7.2Hz, 3H). LCMS[M+H] + 389.

[0452] Example 28: (6R)-6-[4-(3-amino-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]-octane-2- Synthesis of ethyl formate (28)

[0453]

[0454] Step 1: Synthesis of (6R)-6-[4-(3-nitro-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (C39): To a solution of 2-fluoro-3-nitro-pyridine (300 mg, 2.11 mmol) and (6R)-6-piperazin-1-yl-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (677 mg, 2.53 mmol) in MeCN (5.00 mL) was added KCO (875 mg, 6.33 mmol) in one portion at 20°C under a nitrogen atmosphere. The mixture was stirred at 80°C for 12 hours. Water (20 mL) was added to the reaction mixture at 25°C, followed by extraction with EtOAc (3 x 15 mL). The combined organic layers were washed with brine (30 mL), dried over NaSO, filtered, and concentrated under reduced pressure to give a residue. The residue was purified by flash chromatography on silica gel (eluent: 0 to 20% MeOH in DCM) to give (6R)-ethyl 6-[4-(3-nitro-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]-octane-2-carboxylate. LCMS [M+H] + 390.

[0455] Step 2: Synthesis of (6R)-6-[4-(3-amino-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (28): To a solution of (6R)-6-[4-(3-nitro-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (500 mg, 1.28 mmol) in MeOH (10.0 mL) was added Zn (1.67 g, 25.6 mmol) and saturated aqueous NH4Cl solution (10.0 mL) under nitrogen atmosphere. The mixture was stirred at 25°C for 16 hours. The residue was filtered and the filtrate was extracted with DCM (3×10 mL). The combined organic phases were washed with brine (20 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a black oil (360 mg). 320 mg of the black oil was used in the next step without further purification. 40.0 mg of the black oil was purified by preparative HPLC (column: Phenomenex Gemini-NX 80×30 mm×3 μm; conditions: water (10 mM NH4HCO3)-ACN; start B: 27; end B: 57; gradient time (min): 9; 100% B hold time (min): 1.5; flow rate (ml / min): 30; injections: 2) to give (6R)-6-[4-(3-amino-2-pyridyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. 1 H NMR (CDCl3400MHz)δ H =7.75(brd,J=4.4Hz,1H),6.95(dd,J=1.6Hz,7.6Hz,1H),6.88-6.81(m,1H),4.10(q,J=7.2Hz,2H),4.06-3.96(m,1H),3.94-3.8 7(m,1H),3.80-3.75(m,2H),3.74-3.49(m,4H),3.48-2.61(m,6H),2.44-1.73(m,6H),1.70-1.61(m,1H),1.23(t,J=7.2Hz,3H). LCMS[M+H] + 360.

[0456] Examples 29 and 30: (6R)-6-[4-(3-acetylamino-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]- Octane-2-carboxylic acid ethyl ester (29) and (6R)-6-[4-[3-(ethoxycarbonylamino)-2-pyridyl]piperazin-1-yl]-2-aza Synthesis of ethyl spiro[3.4]octane-2-carboxylate (30)

[0457]

[0458] Step to the left: Synthesis of (6R)-6-[4-(3-acetylamino-2-pyridyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (29): Under nitrogen atmosphere, to a solution of (6R)-6-[4-(3-amino-2-pyridyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (50.0 mg, 139 umol) and TEA (42.2 mg, 417 umol) in DCM (3.00 mL) at 20° C. was added CH 3 COCl (13.1 mg, 167 umol) in one portion. The mixture was stirred at 20° C. for 16 hours. The residue was poured onto water (10 mL) and stirred for 5 min. The aqueous phase was extracted with EtOAc (3×10 mL). The combined organic phases were washed with brine (20 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Phenomenex Gemini-NX 80×30 mm×3 μm; conditions: water (10 mM NH4HCO3)-ACN; start B: 23; end B: 53; gradient time (min): 9; 100% B hold time (min): 1.5; flow rate (ml / min): 30; injections: 3) to give (6R)-6-[4-(3-acetylamino-2-pyridyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. 1 H NMR (CDCl3400MHz)δ H =8.53(br d,J=7.2Hz,1H),8.10-8.02(m,1H),7.94-7.56(m,1H),7.07-7.00(m,1H),4.10(q,J=7.2Hz,2H),4.00-3.83(m,2H),3.82-3.74(m,2H ),3.65-3.05(m,4H),3.04-2.28(m,5H),2.23(s,3H),2.20-2.11(m,1H),2.08-1.67(m,4H),1.66-1.60(m,1H),1.24(t,J=7.2Hz,3H). LCMS[M+H] + 402.

[0459] Step to the right: Synthesis of ethyl (6R)-6-[4-[3-(ethoxycarbonylamino)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (30): To a solution of ethyl (6R)-6-[4-[3-(ethoxycarbonylamino)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (50.0 mg, 139 umol) and TEA (42.2 mg, 417 umol) in DCM (3.00 mL) was added ethyl chloroformate (0.160 g, 1.47 mmol) in one portion at 20° C. The mixture was stirred at 20° C. for 16 hours. LCMS showed that the starting material remained. Therefore, more ethyl chloroformate (0.530 g, 4.88 mmol) was added at 20° C. The mixture was stirred for another 4 hours at 20° C. The mixture was poured onto water (10 mL). The aqueous phase was extracted with EtOAc (3×10 mL). The combined organic phases were washed with brine (20.0 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was further purified by preparative HPLC (column: Phenomenex Gemini-NX 80×30 mm×3 μm; conditions: water (10 mM NH₄HCO₃)-ACN; start B: 32; end B: 62; gradient time (min): 9; 100% B hold time (min): 1.5; flow rate (ml / min): 30; injections: 3) to give ethyl (6R)-6-[4-[3-(ethoxycarbonylamino)-2-pyridyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate. 1 H NMR (CDCl3400MHz)δ H =8.40-8.20(m,1H),8.02(dd,J=2.0Hz,4.8Hz,1H),7.26-7.17(m,1H),7.02(dd,J=5.2H z,8.0Hz,1H),4.25(q,J=7.2Hz,2H),4.10(q,J=7.2Hz,2H),3.94-3.69(m,4H),3.07(br t,J=4.8Hz,4H),2.80-2.40(m,5H),2.14(dd,J=7.2Hz,12.8Hz,1H),1.99-1.81(m,3H ),1.81-1.76(m,1H),1.65-1.53(m,1H),1.34(t,J=7.2Hz,3H),1.23(t,J=7.2Hz,3H). LCMS[M+H] + 432.

[0460] Example 31: 6-(4-(3-(Isoxazolyl-3-yl)pyridin-2-yl)piperazin-1-yl)-2-azaspiro[3.4]-octane Synthesis of ethyl 2-oxanecarboxylate (31)

[0461]

[0462] Synthesis of ethyl 6-(4-(3-(isoxazol-3-yl)pyridin-2-yl)piperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylate (31): A solution of ethyl 6-piperazin-1-yl-2-azaspiro[3.4]octane-2-carboxylate (70.0 mg, 262 umol) and 3-(2-fluoro-3-pyridinyl)isoxazole (43.0 mg, 262 umol) in DIPEA (500 uL) and pyridine (250 uL) was stirred at 100° C. for 16 hours. The mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Boston Prime C18 150×30 mm×5 um; mobile phase: [water (0.05% NH3H2O+10 mM NH4HCO3)-ACN]; B%: 40%-70%, 7 min) to give 6-[4-(3-isoxazol-3-yl-2-pyridinyl)-piperazin-1-yl]-2-azaspiro-[3.4]octane-2-carboxylic acid ethyl ester. 1 HNMR (CDCl3400MHz)δ H =8.46(d,J=1.6Hz,1H),8.33(dd,J=2.0Hz,4.8Hz,1H),7.88(dd,J=2.0Hz,7.6Hz ,1H),6.95(dd,J=4.8Hz,7.6Hz,1H),6.87(d,J=1.6Hz,1H),4.09(q,J=6.8Hz,2H) ,3.90-3.75(m,4H),3.18(t,J=4.8Hz,4H),2.63-2.41(m,5H),2.15-2.06(m,1H) ,1.95-1.78(m,3H),1.72-1.67(m,1H),1.57-1.46(m,1H),1.23(t,J=6.8Hz,3H). LCMS [M+H] + 412.

[0463] Example 32: 6-(4-(3-(4-methylisoxazol-3-yl)pyridin-2-yl)piperazin-1-yl)-2-azaspiro [3.4] Synthesis of ethyl octane-2-carboxylate (32)

[0464]

[0465] Synthesis of ethyl 6-(4-(3-(4-methylisoxazol-3-yl)pyridin-2-yl)piperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylate (32): A solution of ethyl 6-piperazin-1-yl-2-azaspiro[3.4]octane-2-carboxylate (100 mg, 374 μmol) and 3-(2-fluoro-3-pyridinyl)-4-methyl-isoxazole (66.6 mg, 374 μmol) in DIPEA (700 μL) and pyridine (350 μL) was stirred at 100° C. for 16 hours. The mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Phenomenex Gemini-NX 80×30 mm×3 um; mobile phase: [water (10 mM NH4HCO3)-ACN]; B%: 32%-62%, 9 min) to give ethyl 6-[4-[3-(4-methylisoxazol-3-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate. 1 HNMR (CDCl3400MHz)δ H =8.34(dd,J=2.0Hz,4.8Hz,1H),8.27-8.23(m,1H),7.60(dd,J=2.0Hz,7.6Hz, 1H),6.92(dd,J=4.8Hz,7.2Hz,1H),4.09(q,J=7.2Hz,2H),3.90-3.72(m,4H), 3.31-3.10(m,4H),2.57-2.29(m,5H),2.12-2.03(m,1H),2.00-1.96(m,3H),1 .94-1.78(m,3H),1.74-1.65(m,1H),1.57-1.45(m,1H),1.23(t,J=7.2Hz,3H). LCMS[M+H] + 426.

[0466] Example 33: (6R)-6-[4-[3-(cyclobutylamino)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]- Synthesis of ethyl octane-2-carboxylate (33)

[0467]

[0468] Synthesis of ethyl (6R)-6-[4-[3-(cyclobutylamino)-2-pyridyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (C40): ethyl (6R)-6-[4-(3-bromo-2-pyridyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (50.0 mg, 118 umol), cyclobutylamine (84.0 mg, 1.18 mmol), t-BuONa (34.1 mg, 354 umol), 2-[bis(3,5-trifluoromethylphenylphosphino)-3,6-dimethoxy]-2,6-dimethylamino-1,1-biphenyl (L7, 8.93 mg, 11.8 umol) and Pd2(dba)3 (10.8 mg, 11.8 umol) in 1,4-dioxane (2.50 mL). The mixture was degassed under vacuum and purged with N2 gas several times. The mixture was then heated to 100 ° C and stirred at 100 ° C for 10 hours. The mixture was cooled to 20 ° C, DCM (20 mL) was added to the mixture, and filtered. The filtrate was concentrated in vacuo. The residue was purified by flash chromatography on silica gel (eluent: 0 to 3.2% MeOH / DCM) to give (6R)-6-[4-[3-(cyclobutylamino)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]-octane-2-carboxylic acid ethyl ester. LCMS [M+H] + 414.

[0469] Step 2: Synthesis of (6R)-6-[4-[3-[cyclobutyl(methyl)amino]-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (33): A mixture of (6R)-6-[4-[3-(cyclobutylamino)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (40.0 mg, 96.7 umol) and NaH (19.3 mg, 484 umol, 60% purity) in DMF (0.500 mL). The mixture was degassed under vacuum and purged with N2 gas several times. The mixture was stirred at 0°C for 0.5 hours. A solution of MeI (16.5 mg, 116 umol) in DMF (0.500 mL) was then added dropwise to the mixture. The mixture was stirred at 0°C for 1.5 hours. HO (5 mL) was added to the mixture, filtered, and concentrated in vacuo to afford the crude product. The crude product was purified by preparative HPLC (column: Phenomenex Gemini-NX 80×40 mm×3 μm; mobile phase: [water (0.05% NH 3 H 2 O)-ACN]; B%: 50%-80%, 8 min). The fractions were then concentrated in vacuo and freeze-dried to afford ethyl (6R)-6-[4-[3-[cyclobutyl-(methyl)amino]-2-pyridyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate.1 H NMR (MeOD 400 MHz) δ H =7.80(dd,J=1.2Hz,4.8Hz,1H),7.10(dd,J=1.2Hz,7.6Hz,1H),6.85(dd,J=4.8Hz,7.6Hz,1H),4.08(q,J=7.2Hz,2H),3.95-3.75(m,5H), 3.66-3.39(m,4H),2.75-2.60(m,8H),2.28-2.15(m,3H),2.03-1.85(m,5H),1.83-1.70(m,3H),1.64-1.55(m,1H),1.24(t,J=6.8Hz,3H). LCMS[M+H] + 428.

[0470] Examples 34 and 235: (6R)-6-[4-[3-(5-oxopyrrolidin-3-yl)-2-pyridinyl]piperazin-1-yl]-2- Azaspiro[3.4]octane-2-carboxylic acid ethyl ester (34) and (6R)-6-[4-[3-(1-methyl-5-oxo-pyrrolidin-3-yl)-2-pyrrolidone]- Synthesis of ethyl [pyridyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (235)

[0471]

[0472] Step 1: Synthesis of (6R)-6-[4-[3-[(E)-3-ethoxy-3-oxo-prop-1-enyl]-2-pyridinyl]-piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (C41): To (6R)-6-[4-(3-bromo-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (500 mg, 1.18 mmol), (E)-3-(4 A mixture of [1,1'-bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) (Pd-118, 76.9 mg, 118 umol) and K2CO3 (326 mg, 2.36 mmol) in H2O (2.00 mL) and 1,4-dioxane (10.0 mL) was added. The mixture was degassed under reduced pressure and purged with N2 gas several times. The reaction mixture was stirred at 85°C for 16 hours. The mixture was concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel (eluent: 0 to 5% MeOH / DCM) to give (6R)-6-[4-[3-[(E)-3-ethoxy-3-oxo-prop-1-enyl]-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. LCMS [M+H] + 443.

[0473] Step 2: Synthesis of (6R)-6-[4-[3-[3-ethoxy-1-(nitromethyl)-3-oxo-propyl]-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (C42): To a mixture of (6R)-6-[4-[3-[(E)-3-ethoxy-3-oxo-prop-1-enyl]-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (300 mg, 678 umol) in CH3NO2 (1.50 mL, 27.8 mmol) was added DBU (206 mg, 1.36 mmol) dropwise at 0°C. The mixture was then stirred at 20°C for 12 hours. The mixture was concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel (0 to 10% MeOH / DCM as eluent) to give (6R)-6-[4-[3-[3-ethoxy-1-(nitromethyl)-3-oxo-propyl]-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. LCMS [M+H] + 504.

[0474] Step 3: Synthesis of (6R)-6-[4-[3-(5-oxopyrrolidin-3-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (34): To a mixture of (6R)-6-[4-[3-[3-ethoxy-1-(nitromethyl)-3-oxo-propyl]-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (160 mg, 318 umol) in EtOH (8.00 mL) was added 10% wet Pd / C (30 mg) at 20° C. under N atmosphere. The mixture was then degassed under reduced pressure and purged with H gas several times. The mixture was then stirred at 60° C. under H pressure (50 psi) for 24 h. The residue was purified by flash chromatography on silica gel (eluent: 0 to 15% MeOH / DCM) and preparative HPLC (column: Boston Prime C18 150×30 mm×5 um; mobile phase: [water (NH3H2O+NH4HCO3)-ACN]; B%: 20%-50%, 7 min) to give (6R)-6-[4-[3-(5-oxopyrrolidin-3-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. 1 H NMR (CDCl3400MHz)δ H=8.24(dd,J=2.0Hz,4.8Hz,1H),7.59(dd,J=1.2Hz,7.2Hz,1H),7.00(dd,J=4.8Hz,7.6Hz, 1H),5.97(s,1H),4.15-3.99(m,3H),3.93-3.76(m,5H),3.32(dd,J=6.8Hz,9.6Hz,1H),3. 19-3.02(m,4H),2.83-2.74(m,1H),2.73-2.52(m,5H),2.40(dd,J=8.0Hz,16.8Hz,1H),2. 13(dd,J=7.2Hz,12.8Hz,1H),2.02-1.76(m,4H),1.65-1.56(m,1H),1.23(t,J=6.8Hz,3H). LCMS[M+H] + 428.

[0475] Step 4: Synthesis of (6R)-6-[4-[3-(1-methyl-5-oxo-pyrrolidin-3-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (235): To a mixture of (6R)-6-[4-[3-(5-oxopyrrolidin-3-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (50.0 mg, 117 umol) in DMF (1.50 mL) was added NaH (14.0 mg, 351 umol, 60% purity) at 0° C. and stirred at 0° C. for 30 minutes under N2 atmosphere. Then, CH3I (16.6 mg, 117 umol) was added to the mixture at 0° C. and the reaction was warmed to 20° C. and stirred at 20° C. for 1 hour. Then, H2O (5 mL) was added dropwise to the mixture, and the mixture was extracted with DCM (3×5 mL) and concentrated under reduced pressure to provide a crude material. The crude material was purified by preparative HPLC (column: Welch Xtimate C18 150×30 mm×5 um; mobile phase: [water (NH3H2O+NH4HCO3)-ACN]; B%: 30%-60%, 7 min) to give (6R)-6-[4-[3-(1-methyl-5-oxo-pyrrolidin-3-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. 1 H NMR (CDCl3400MHz)δ H=8.24(dd,J=2.0Hz,4.8Hz,1H),7.52(dd,J=2.0Hz,8.0Hz,1H),7.00(dd,J=4.4 Hz,7.2Hz,1H),4.09(q,J=7.2Hz,2H),3.94-3.76(m,6H),3.30(dd,J=6.0Hz,9.6 Hz,1H),3.25-2.95(m,4H),2.93-2.82(m,4H),2.81-2.50(m,5H),2.45(dd,J=6 .4Hz,16.8Hz,1H),2.21-2.10(m,1H),1.99-1.64(m,5H),1.24(t,J=7.2Hz,3H). LCMS[M+H] + 442.

[0476] Example 35: (R)-6-(4-(3-(1-methylpiperidin-4-yl)pyridin-2-yl)piperazin-1-yl)-2-azaspiro [3.4] Synthesis of ethyl octane-2-carboxylate (35)

[0477]

[0478] Step 1: Synthesis of (R)-6-(4-(1'-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridyl]-2-yl)-piperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (C43): Under N2 atmosphere, (6R)-6-[4-(3-bromo-2-pyridyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (100 mg, 236 μmol), 1-methyl-4-(1'-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridyl]-2-yl)-piperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylate was added at 20°C. To a mixture of -(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydro-2H-pyridine (105 mg, 472 umol) and Na2CO3 (75.1 mg, 709 umol) in 1,4-dioxane (4.00 mL) and H2O (1.00 mL) were added di-tert-butyl(cyclopentyl)phosphine; dichloropalladium; and iron (Pd(dtbpf)Cl2, 15.4 mg, 23.6 umol) in one portion. The mixture was then degassed under vacuum and purged with N2 gas several times. The mixture was stirred at 80°C for 16 hours. The reaction mixture was concentrated. The residue was purified by flash silica gel chromatography (0 to 10% MeOH / DCM as eluent) to give (6R)-6-[4-[3-(1-methyl-3,6-dihydro-2H-pyridin-4-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro-[3.4]octane-2-carboxylic acid ethyl ester. LCMS [M+H] + 440.

[0479] Step 2: Synthesis of (R)-6-(4-(3-(1-methylpiperidin-4-yl)pyridin-2-yl)piperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (35): To a solution of (6R)-6-[4-[3-(1-methyl-3,6-dihydro-2H-pyridin-4-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (100 mg, 227 umol) in THF (5.00 mL) was added Pd / C (wet) (10.0 mg, 10.0% purity) under N2 atmosphere. The suspension was degassed and purged with H2 gas three times. The mixture was stirred at 20°C under H2 gas (15 psi) for 16 h. The reaction mixture was filtered and the filtrate was concentrated. The residue was purified by preparative HPLC (column: Welch Xtimate C18 150×30 mm×5 um; mobile phase: [water (NH3H2O+NH4HCO3)-ACN]; B%: 32%-62%, 7 min) to give (6R)-6-[4-[3-(1-methyl-4-piperidinyl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. 1 H NMR (CDCl3400MHz)δ H =8.17(dd,J=1.6Hz,4.8Hz,1H),7.51(dd,J=1.6Hz,7.6Hz,1H),6.94(dd,J=4.8Hz,7.2Hz,1H) ,4.11(q,J=7.2Hz,2H),3.94-3.84(m,2H),3.84-3.78(m,2H),3.12(t,J=4.8Hz,4H),3.02(br d,J=11.2Hz,2H),2.87-2.76(m,1H),2.63(br s,5H),2.36(s,3H),2.16(dd,J=6.8Hz,12.8Hz,1H),2.11-2.02(m,2H),2.00-1.90(m,2H),1. 86(brdd,J=5.2Hz,13.6Hz,1H),1.80-1.69(m,5H),1.64-1.55(m,1H),1.25(t,J=7.2Hz,3H). LCMS[M+H] + 442.

[0480] Examples 36, 236 and 237: 6-(4-(5-fluoro-3-(pyrazin-2-yl)pyridin-2-yl)piperidin-1-yl)-2-azepine Spiro[3.4]octane-2-carboxylic acid ethyl ester (36), (6R)-6-[4-(5-fluoro-3-pyrazin-2-yl-2-pyridinyl)-1-piperidinyl]-2- Azaspiro[3.4]octane-2-carboxylic acid ethyl ester (236) and (6S)-6-[4-(5-fluoro-3-pyrazin-2-yl-2-pyridinyl)-1-piperidin Synthesis of ethyl]-2-azaspiro[3.4]octane-2-carboxylate (237)

[0481]

[0482] Step 1: Synthesis of ethyl 6-(4-(5-fluoro-3-(pyrazin-2-yl)pyridin-2-yl)piperidin-1-yl)-2-azaspiro[3.4]octane-2-carboxylate (36): To a mixture of 2-[5-fluoro-2-(4-piperidinyl)-3-pyridinyl]pyrazine (90.0 mg, 348 umol) and ethyl 6-oxo-2-azaspiro[3.4]octane-2-carboxylate (75.6 mg, 383 umol) in DCE (2.00 mL) was added TEA (176 mg, 1.74 mmol) and the mixture was stirred at 20° C. for 2 hours. NaBH(OAc) 3 (221 mg, 1.05 mmol) and HOAc (20.9 mg, 348 umol) were then added at 20° C. The mixture was stirred at 20° C. for 18 hours under N 2 atmosphere. The mixture was concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Welch Xtimate C18 100×40 mm×3 um; mobile phase: [water (TFA)-ACN]; B%: 5%-35%, 8 min) to give ethyl 6-[4-(5-fluoro-3-pyrazin-2-yl-2-pyridinyl)-1-piperidinyl]-2-azaspiro[3.4]octane-2-carboxylate. 1 H NMR (CDCl3400MHz)δ H =8.63(s,1H),8.80-8.75(m,1H),8.70(d,J=2.4Hz,1H),8.60(d,J=2.8Hz, 1H),7.79(dd,J=2.8Hz,9.2Hz,1H),4.09(q,J=7.2Hz,2H),3.97-3.83(m,4 H),3.65(brd,J=12.0Hz,2H),3.58-3.53(m,1H),3.04-2.94(m,2H),2.48- 2.40(m,1H),2.35-1.98(m,9H),1.85-1.76(m,1H),1.22(t,J=7.2Hz,3H). 19 F NMR (CDCl3400MHz)δ F =-77.130,-131.279. LCMS[M+H] + 440.

[0483] Step 2: Synthesis of (6R)-6-[4-(5-fluoro-3-pyrazin-2-yl-2-pyridinyl)-1-piperidinyl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (236) and (6S)-6-[4-(5-fluoro-3-pyrazin-2-yl-2-pyridinyl)-1-piperidinyl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (237): 6-[4-(5-fluoro-3-pyrazin-2-yl-2-pyridinyl)-1-piperidinyl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (100 mg, 228 μmol) was purified by SFC (column: DAICEL CHIRALPAK IG (250mm×30mm, 10um); mobile phase: [CO2-MeOH (0.1% NH3H2O)]; B%: 55%-55%, 9min) purification to obtain (6R)-6-[4-(5-fluoro-3-pyrazin-2-yl-2-pyridinyl)-1-piperidinyl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. Peak 1, 1 HNMR (CDCl3400MHz)δ H =8.73-8.68(m,2H),8.64(d,J=2.4Hz,1H),8.53(d,J=2.8Hz,1H),7.42(dd,J=2.8Hz,8.4Hz,1H),4.09(q,J=7.2Hz,2H),3.94-3.78(m,2H) ,3.78-3.72(m,2H),3.15-2.98(m,2H),2.92-2.75(m,1H),2.67-2.41(m,1H),2.17-2.02(m,3H),2.00-1.65(m,9H),1.23(t,J=7.2Hz,3H). LCMS[M+H] + 440. and (6S)-6-[4-(5-fluoro-3-pyrazin-2-yl-2-pyridinyl)-1-piperidinyl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. Peak 2, 1 H NMR (CDCl3400MHz)δ H =8.71(s,2H),8.64(d,J=2.0Hz,1H),8.53(d,J=3.2Hz,1H),7.48-7.35(m,1H),4.09(q,J=7.2Hz,2H),3.94-3.79(m, 2H), 3.76 (s, 2H), 3.31-2.71 (m, 3H), 2.70-2.42 (m, 1H), 2.30-2.05 (m, 3H), 2.03-1.64 (m, 9H), 1.23 (t, J = 7.2Hz, 3H). LCMS[M+H] + 440.

[0484] Example 37: (6R)-6-[4-[5-fluoro-3-(4-hydroxybut-1-ynyl)-2-pyridinyl]piperazin-1-yl]-2-nitropropene Synthesis of Heterospiro[3.4]octane-2-carboxylic Acid Ethyl Ester (238)

[0485]

[0486] Synthesis of (6R)-6-[4-[5-fluoro-3-(4-hydroxybut-1-ynyl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (238): At 20°C, (6R)-6-[4-(3-bromo-5-fluoro-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (100 mg, 22 To a mixture of toluene (2.00 mL) and H2O (1.00 mL) was added Pd(PPh3)2Cl2 (15.9 mg, 22.7 umol), but-3-yn-1-ol (31.8 mg, 453 umol, 34.3 uL), CuI (4.32 mg, 22.7 umol) and TEA (45.7 mg, 453 umol). The mixture was then degassed under vacuum and purged with N2 gas several times. The reaction mixture was heated to 110 ° C and stirred at 110 ° C for 12 hours under a N2 gas atmosphere. The mixture was cooled to 20 ° C and concentrated under reduced pressure. The residue was purified by silica gel chromatography (eluent: 0 to 10% MeOH / DCM) and further purified by preparative HPLC (column: Welch Xtimate C18 150×25 mm×5 um; mobile phase: [water(NH 3 H 2 O)-ACN]; B%: 22%-52%, 8 min) to give (6R)-6-[4-[5-fluoro-3-(4-hydroxybut-1-ynyl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. 1 HNMR (CDCl3400MHz)δ H =8.03(d,J=3.2Hz,1H),7.35(dd,J=2.8Hz,8.4Hz,1H),4.10(q,J=7.2Hz,2H),3.92-3.76(m,6H),3.50(br s,4H),2.74(t,J=6.4Hz,,2H),2.62(br s, 5H), 2.14 (dd, J = 6.8Hz, 12.0Hz, 1H), 2.02-1.71 (m, 5H), 1.24 (t, J = 7.2Hz, 3H). LCMS[M+H] + 431.

[0487] Example 38: (6R)-6-[4-(3-ethynyl-5-fluoro-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane Ethyl 2-oxanecarboxylate (239) and (6R)-6-[4-[5-fluoro-3-(1H-triazol-4-yl)-2-pyridinyl]piperazin-1-yl]-2-nitropropane Synthesis of Heterospiro[3.4]octane-2-carboxylic Acid Ethyl Ester (240)

[0488]

[0489] Step 1: Synthesis of (R)-6-(4-(5-fluoro-3-formylpyridin-2-yl)piperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (C44): A mixture of 2-chloro-5-fluoro-pyridine-3-carbaldehyde (1.00 g, 6.27 mmol) and KCO (866 mg, 6.27 mmol) was stirred in 1,4-dioxane (40.0 mL). Then, (6R)-6-piperazin-1-yl-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (2.18 g, 8.15 mmol) was added and the mixture was stirred at 20° C. for 16 hours. The mixture was diluted with EtOAc (100 mL), filtered, concentrated, and purified by flash silica gel chromatography (0 to 10% MeOH / DCM as eluent) to give (R)-ethyl 6-(4-(5-fluoro-3-formylpyridin-2-yl)piperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylate. LCMS [M+H] + 391.

[0490] Step 2: Synthesis of (R)-6-(4-(3-ethynyl-5-fluoropyridin-2-yl)piperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (239): A mixture of (6R)-6-[4-(5-fluoro-3-formyl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (100 mg, 256 umol) and KCO (70.8 mg, 512 umol) was stirred in MeOH (3.00 mL). Then, a solution of 1-diazo-1-dimethoxyphosphoryl-propan-2-one (54.1 mg, 281 umol) in MeOH (3.00 mL) was added and the mixture was stirred at 25° C. for 5 hours. The mixture was diluted with water (3 mL) and concentrated. To the resulting residue was added water (5 mL) again, and the aqueous phase was extracted with EtOAc (10 mL × 3). The combined organic layers were washed with brine (10 mL), dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified by preparative HPLC (column: Waters xbridge 150 × 25 mm 10 μm; mobile phase: [water (NH4HCO3)-ACN]; B%: 36%-66%, 8 min) to give (6R)-6-[4-(3-ethynyl-5-fluoro-2-pyridyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. 1 H NMR (CDCl3400MHz)δ H=8.09(d,J=2.8Hz,1H),7.45(dd,J=3.2Hz,8.0Hz,1H),4.12(q,J=7.2Hz,2H),3 .95-3.90(m,1H),3.89-3.85(m,1H),3.84-3.78(m,2H),3.62-3.52(m,4H),3.51 (s,1H),2.75-2.55(m,5H),2.17(dd,J=6.8Hz,12.4Hz,1H),2.03-1.91(m,2H), 1.91-1.84(m,1H),1.82-1.71(m,1H),1.69-1.62(m,1H),1.26(t,J=6.8Hz,3H). LCMS [M+H] + 387.

[0491] Step 3: Synthesis of (6R)-6-[4-[5-fluoro-3-(1H-triazol-4-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (240): A mixture of (6R)-6-[4-(3-ethynyl-5-fluoro-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]-octane-2-carboxylic acid ethyl ester (48.0 mg, 124 umol), TMSN3 (71.5 mg, 621 umol), CuSO4·5H2O (3.10 mg, 12.4 umol) and sodium ascorbate (4.92 mg, 24.8 umol) in t-BuOH (4.00 mL) and H2O (1.00 mL) was degassed and purged with N2 gas three times. The mixture was then stirred at 80° C. under N₂ atmosphere for 18 hours. The mixture was quenched with water (10 mL) and extracted with EtOAc (20 mL×3). The combined organic phases were washed with brine (10 mL), dried over Na₂SO₄, and concentrated. The residue was purified by preparative HPLC (column: Welch Ultimate C18 150×25 mm×5 μm; mobile phase: [water (FA)-ACN]; B%: 6%-36%, 10 min) to give (6R)-6-[4-[5-fluoro-3-(1H-triazol-4-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. 1 H NMR (CDCl3400MHz)δ H=8.23(br s,1H),8.18-8.09(m,1H),7.90-7.80(m,1H),4.10(q,J=6.8Hz,2H),3.97-3.74(m,4H),3.35- 3.18(m,4H),3.00-2.72(m,5H),2.26-2.14(m,1H),2.10-1.74(m,5H),1.23(t,J=7.2Hz,3H). LCMS[M+H] + 430.

[0492] Example 39: (6R)-6-[4-[3-(1,4-dimethylpyrazol-3-yl)-2-pyridinyl]piperazin-1-yl]-2-nitrogen Heterospiro-[3.4]octane-2-carboxylic acid ethyl ester (241) and (6R)-6-[4-[3-(2,4-dimethylpyrazol-3-yl)-2-pyridinyl] Synthesis of ethyl piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (242)

[0493]

[0494] Step 1: Synthesis of (R)-ethyl 6-(4-(3-propionylpyridin-2-yl)piperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylate (C45): Under N2, a solution of 1-(2-chloro-3-pyridinyl)propan-1-one (500 mg, 2.95 mmol) in DIPEA (1.50 mL) and pyridine (0.500 mL) was mixed with ethyl (6R)-6-piperazin-1-yl-2-azaspiro[3.4]octane-2-carboxylate (828 mg, 3.10 mmol) in one portion at 20°C. The mixture was heated to 130°C and stirred at 130°C for 12 h. The reaction mixture was then poured into an aqueous HCl solution (2 M, 30 mL) at 20° C., extracted with EtOAc (3×30 mL), and the combined organic layers were washed with brine (50 mL), dried over Na SO , filtered, and concentrated to give a residue. The residue was purified by flash silica gel chromatography (eluent: 0 to 10% MeOH in DCM) to give ethyl (6R)-6-[4-(3-propionyl-2-pyridyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate. 1 H NMR (CDCl3400MHz)δ H=8.27(dd,J=1.6Hz,4.8Hz,1H),7.63(dd,J=1.6Hz,7.2Hz,1H),6.82(dd,J=5.6Hz,7.2Hz,1H),4.11(q,J=7.2Hz,2H),3.93-3.71(m,4H),3.34(br t,J=4.8Hz,4H),2.96(q,J=7.2Hz,2H),2.65-2.53(m,5H),2.12(dd,J=7.2Hz,12.8Hz,1H),2.00- 1.80(m,3H),1.79-1.67(m,1H),1.55-1.50(m,1H),1.23(t,J=7.6Hz,3H),1.16(t,J=7.2Hz,3H). LCMS[M+H] + 401.

[0495] Step 2: Synthesis of (R,Z)-ethyl 6-(4-(3-(3-(dimethylamino)-2-methacryloyl)pyridin-2-yl)piperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylate (C46): Under N2, a solution of ethyl (6R)-6-[4-(3-propionyl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (100 mg, 250 umol) in MeCN (2.00 mL) was mixed with 1,1-dimethoxy-N,N-dimethyl-methylamine (149 mg, 1.25 mmol) in one portion at 20°C. The mixture was heated to 115°C and stirred at 115°C for 30 hours. The reaction mixture was concentrated to give (6R)-6-[4-[3-[(Z)-3-(dimethylamino)-2-methyl-prop-2-enoyl]-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. The crude product was used directly in the next step without further purification. LCMS [M+H] + 456.

[0496] Step 3: Synthesis of (6R)-6-[4-[3-(1,4-dimethylpyrazol-3-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro-[3.4]octane-2-carboxylic acid ethyl ester (241) and (6R)-6-[4-[3-(2,4-dimethylpyrazol-3-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (242): In N To a solution of (6R)-6-[4-[3-[(Z)-3-(dimethylamino)-2-methyl-prop-2-enoyl]-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (400 mg, 878 umol) in MeOH (5.00 mL) was added methylhydrazine hydrate (2.77 g, 17.3 mmol) in a single portion under atmospheric pressure at 20° C. The mixture was heated to 70° C. and stirred at 70° C. for 12 hours. The reaction mixture was poured into water (20.0 mL) at 25° C. and then extracted with EtOAc (3×15 mL). The combined organic layers were washed with brine (30 mL), dried over Na 2 SO 4 , filtered and concentrated under reduced pressure to provide a residue. The residue was purified by flash silica gel chromatography (eluent: 0 to 10% MeOH in DCM) to give a mixture of products. The crude product was purified by SFC ([0.1% NH3H2O ​​EtOH]; B%: 30%-30%, 9 min) to give (6R)-6-[4-[3-(1,4-dimethylpyrazol-3-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. Peak 1, 1 HNMR (CDCl3400MHz)δ H =8.31(dd,J=2.0,4.8Hz,1H),7.43-7.26(m,2H),6.91(dd,J=5.2,7.6Hz,1H),4.09(q,J=7.2Hz,2H),3.96-3.51(m,7H),3.13(br s,4H),2.80-2.14(m,5H),2.13-2.04(m,1H),1.97(s,3H),1.94-1.73(m,4H),1.56-1.47(m,1H),1.23(t,J=7.2Hz,3H). LCMS[M+H] + 439. and (6R)-6-[4-[3-(2,4-dimethylpyrazol-3-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. Peak 2, 1 HNMR (CDCl3400MHz)δ H=8.22(dd,J=2.0,5.2Hz,1H),7.52(dd,J=2.0,7.2Hz,1H),7.17(s,1H),6.83(d d,J=4.8,7.6Hz,1H),4.07(q,J=7.2Hz,2H),3.90-3.64(m,7H),3.16(t,J=4.8H z,4H),2.54-2.30(m,5H),2.10-2.02(m,1H),1.97(s,3H),1.96-1.94(m,1H),1 .90-1.80(m,2H),1.72-1.60(m,1H),1.56-1.44(m,1H),1.21(t,J=7.2Hz,3H). LCMS [M+H] + 439.

[0497] Example 40: 6(R)-(4-(3-(1-methyl-1H-pyrazol-4-yl)pyridin-2-yl)piperazin-1-yl)-2-azepine Synthesis of Spiro[3.4]octane Hydrochloride (243)

[0498]

[0499] Step 1: Synthesis of (R)-6-(4-(3-(1-methyl-1H-pyrazol-4-yl)pyridin-2-yl)piperazin-1-yl)-2-azaspiro[3.4]octane hydrochloride (243): To a solution of tert-butyl (6R)-6-[4-[3-(1-methylpyrazol-4-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (250 mg, 552 umol) in 2-MeTHF (2.00 mL) was added HCl / H2O (6 M in H2O, 0.900 mL) in one portion at 20°C. The mixture was stirred at 20°C for 3 hours. The reaction mixture was extracted with EtOAc (3×5 mL). The aqueous phase was concentrated under reduced pressure. The residue was freeze-dried to give (6R)-6-[4-[3-(1-methylpyrazol-4-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane. 1 H NMR (methanol-d4400MHz)δ H =8.37(s,1H),8.35-8.20(m,2H),8.08(s,1H),7.53(dd,J=6.4Hz,7.6Hz,1H),4.35-3.77(m,10H),3.75 -3.57(m,4H),3.55-3.37(m,2H),2.63(dd,J=8.4Hz,13.6Hz,1H),2.40-2.21(m,3H),2.12-1.92(m,2H). LCMS[M+H] + 353.

[0500] Example 41: (R)-6-(4-(6-fluoro-5-hydroxy-3-(pyrazin-2-yl)pyridin-2-yl)piperazin-1-yl)-2-nitrogen Synthesis of Heterospiro[3.4]octane-2-carboxylic Acid Ethyl Ester (244)

[0501]

[0502] Step 1: Synthesis of 5-bromo-6-fluoro-2-nitropyridine-3-ol (C48): Concentrated aliquots of HSO (13.0 mL) and KNO (470 mg, 4.65 mmol) were mixed in one portion at room temperature and stirred at room temperature for 30 min. Then, 5-bromo-6-fluoro-pyridin-3-ol (500 mg, 2.60 mmol) was added in one portion and the mixture was stirred at room temperature for 16 h. The mixture was slowly poured into ice water (50 mL) and extracted with EtOAc (3×50 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous NaSO, filtered and concentrated under reduced pressure to give 5-bromo-6-fluoro-2-nitropyridine-3-ol. 1 HNMR (CDCl3400MHz)δ H =10.21(s,1H),8.00(d,J=6.4Hz,1H).

[0503] Step 2: Synthesis of 3-(benzyloxy)-5-bromo-6-fluoro-2-nitropyridine (C49): To a solution of 5-bromo-6-fluoro-2-nitro-pyridin-3-ol (530 mg, 2.24 mmol) in DMF (5.00 mL) was added NaH (134 mg, 3.35 mmol, 60% purity) in one portion at 0°C under N2 and stirred at 0°C for 30 min. Next, BnBr (765 mg, 4.47 mmol) was added in one portion at 0°C and the mixture was then warmed to room temperature and stirred at room temperature for 16 hours. The reaction mixture was poured into water (30 mL) and then extracted with EtOAc (3×30 mL). The combined organic layers were washed with brine (30 mL), dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to give a residue. The residue was purified by flash chromatography on silica gel (eluent: 0 to 20% EtOAc in petroleum ether) to give 5-benzyloxy-3-bromo-2-fluoro-6-nitro-pyridine. 1 HNMR (CDCl3400MHz)δ H =7.86(d,J=6.4Hz,1H),7.48-7.31(m,5H),5.25(s,2H). LCMSm / z[M+H] + 327,329.

[0504] Step 3: Synthesis of (R)-6-(4-(5-benzyloxy)-3-bromo-6-nitropyridin-2-yl)piperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (C50): A solution of 5-benzyloxy-3-bromo-2-fluoro-6-nitro-pyridine (500 mg, 1.53 mmol) in DMSO (20.0 mL) was mixed with (6R)-6-piperazin-1-yl-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (409 mg, 1.53 mmol) and KCO (634 mg, 4.59 mmol) in one portion. The mixture was stirred at room temperature for 12 hours. The mixture was then poured into water (30 mL), extracted with EtOAc (3×20 mL) and washed with brine (50 mL). Next, the mixture was dried over anhydrous Na2SO4, filtered, and concentrated to give a residue. The residue was purified by flash silica gel chromatography (eluent: 0 to 10% MeOH / DCM) to give (6R)-6-[4-(5-benzyloxy-3-bromo-6-nitro-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. LCMS m / z [M+H] + 574,576.

[0505] Step 4: Synthesis of (R)-6-(4-(5-(benzyloxy)-6-nitro-3-(pyrazin-2-yl)pyridin-2-yl)piperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (C51): A solution of (6R)-6-[4-(5-benzyloxy-3-bromo-6-nitro-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (250 mg, 435 μmol) and tributyl(pyrazin-2-yl)stannane (193 mg, 522 μmol) in toluene (10.0 mL) was mixed with CuI (8.29 mg, 43.5 μmol) and Pd(PPh3)4 (25.1 mg, 21.8 μmol) at once. The mixture was degassed under vacuum and purged with N2 gas three times and stirred at 110 ° C for 16 hours. Then, the reaction mixture was filtered and concentrated to give a residue. The residue was purified by flash silica gel chromatography (eluent: 0 to 10% MeOH / DCM) to give (6R)-6-[4-(5-benzyloxy-6-nitro-3-pyrazin-2-yl-2-pyridyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. LCMSm / z[M+H] + 574.

[0506] Step 5: Synthesis of (R)-ethyl 6-(4-(5-(benzyloxy)-6-fluoro-3-(pyrazin-2-yl)pyridin-2-yl)piperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylate (C52): To a solution of ethyl (6R)-6-[4-(5-benzyloxy-6-nitro-3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (80.0 mg, 139 μmol) in DMSO (2.00 mL) was added a solution of TBAF in THF (1 M, 1.00 mL) in one portion. The mixture was stirred at room temperature for 16 h. The mixture was poured into water (20 mL) and extracted with EtOAc (3×10 mL). The combined organic layers were washed with water (2 x 20 mL), brine (20 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to yield a residue. The residue was purified by flash silica gel chromatography (eluent: 0 to 12% MeOH / DCM) to yield (6R)-6-[4-(5-benzyloxy-6-fluoro-3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. LCMS m / z [M+H] + 547.

[0507] Step 6: Synthesis of (R)-ethyl 6-(4-(6-fluoro-5-hydroxy-3-(pyrazin-2-yl)pyridin-2-yl)piperazin-1-yl)-2-azaspiro[3.4]octane-2-carboxylate (244): To a solution of ethyl (6R)-6-[4-(5-benzyloxy-6-fluoro-3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate (70.0 mg, 128.06 μmol) in THF (10.0 mL) was added dry Pd / C (20 mg, 10% w / w) in one portion. The mixture was degassed and purged with H gas three times. The mixture was stirred at 30° C. under H gas (40 psi) for 16 h. The reaction mixture was filtered and concentrated to give a residue. The residue was purified by flash silica gel chromatography (eluent: 0 to 10% MeOH / DCM) and further purified by preparative HPLC (column: Phenomenex C18, 80×40 mm×3 um; mobile phase: [water(NH3H2O+NH4HCO3)-ACN]; gradient: 19%-49% B over 8 min) to give (6R)-6-[4-(6-fluoro-5-hydroxy-3-pyrazin-2-yl-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. 1 HNMR (CD3CN400MHz)δ H=9.27(d,J=1.6Hz,1H),8.62(dd,J=1.6Hz,2.4Hz,1H),8.46(d,J=2.4Hz,1 H),7.64(d,J=10.8Hz,1H),4.00(q,J=7.2Hz,2H),3.82-3.61(m,4H),2.92 (t,J=4.8Hz,4H),2.61-2.38(m,5H),2.06-2.01(m,1H),1.88-1.76(m,3H) ,1.62(dd,J=9.2Hz,12.8Hz,1H),1.50-1.38(m,1H),1.17(t,J=7.2Hz,3H). 19 F NMR (CD3CN 400MHz)δ F =-89.28. LCMS m / z [M+H] + 457.

[0508] Example 42: (R)-6-(4-fluoro-4-(5-fluoro-3-(pyrazin-2-yl)pyridin-2-yl)piperidin-1-yl)-2-azepine Spiro[3.4]octane-2-carboxylic acid ethyl ester (245) and (S)-6-(4-fluoro-4-(5-fluoro-3-(pyrazin-2-yl)pyridin-2-yl)piperidin- Synthesis of ethyl 1-amino-2-azaspiro[3.4]octane-2-carboxylate (246)

[0509]

[0510] Step 1: Synthesis of tert-butyl 4-(3-bromo-5-fluoro-2-pyridinyl)-4-hydroxy-piperidine-1-carboxylate (C53): Under N2, a mixture of 2,3-dibromo-5-fluoro-pyridine (3.50 g, 13.7 mmol) in DCM (35.0 mL) was mixed with n-BuLi (2.5 M, 6.09 mL) in n-hexane at -70°C and stirred for 10 min at -70°C. Then, a solution of tert-butyl 4-oxopiperidine-1-carboxylate (2.19 g, 11.0 mmol) in DCM (20.0 mL) was added dropwise to the mixture at -70°C and the mixture was stirred at -70°C for 1 hour. The reaction mixture was quenched with saturated aqueous NH4Cl solution (50 mL) and extracted with DCM (3 x 20 mL). The combined organic layers were washed with brine (50 mL), dried over anhydrous NaSO, filtered and concentrated in vacuo. The residue was purified by flash silica gel chromatography (0 to 13% EtOAc / petroleum ether as eluent) to afford tert-butyl 4-(3-bromo-5-fluoro-2-pyridinyl)-4-hydroxy-piperidine-1-carboxylate. 1 HNMR (CDCl3400MHz)δ H=8.39(d,J=2.4Hz,1H),7.75(dd,J=2.4Hz,7.2Hz,1H),4.21-3.97(m,2H),3.45-3.15(m,2H),2.85-2.73(m,2H),1.49(s,9H),1.45-1.36(m,2H). LCMSm / z[M+H-100] + 275,277.

[0511] Step 2: Synthesis of tert-butyl 4-(3-bromo-5-fluoro-2-pyridinyl)-4-fluoro-piperidine-1-carboxylate (C54): A mixture of tert-butyl 4-(3-bromo-5-fluoro-2-pyridinyl)-4-hydroxy-piperidine-1-carboxylate (700 mg, 1.87 mmol) in DCM (7.00 mL) was mixed with DAST (756 mg, 4.69 mmol) at 0° C. and stirred at 25° C. for 12 hours. The mixture was added to saturated aqueous NaHCO 3 solution (50 mL) and extracted with DCM (3×10 mL). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na 2 SO 4 , filtered, and concentrated in vacuo. The residue was purified by flash silica gel chromatography (eluent: 0 to 7% EtOAc / petroleum ether) to give tert-butyl 4-(3-bromo-5-fluoro-2-pyridinyl)-4-fluoro-piperidine-1-carboxylate. LCMS m / z[M+H-56] + 321,323.

[0512] Step 3: Synthesis of 4-fluoro-4-[5-fluoro-3-(1-methylpyrazol-4-yl)-2-pyridinyl]-piperidine-1-carboxylic acid tert-butyl ester (C55): A mixture of 4-(3-bromo-5-fluoro-2-pyridinyl)-4-fluoro-piperidine-1-carboxylic acid tert-butyl ester (340 mg, 901 μmol) and 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrazole (225 mg, 1.08 mmol) in 1,4-dioxane (5.00 mL) and H2O (0.700 mL) was mixed with Pd(dppf)Cl2 (66.0 mg, 90.1 μmol) and Na2CO3 (191 mg, 1.80 mmol). The mixture was degassed under vacuum and purged with N gas three times and stirred for 12 hours at 100° C. The mixture was concentrated and purified by flash silica gel chromatography (eluent: 0 to 49% EtOAc / petroleum ether) to give tert-butyl 4-fluoro-4-[5-fluoro-3-(1-methylpyrazol-4-yl)-2-pyridinyl]piperidine-1-carboxylate. 1 H NMR (CDCl3400MHz)δ H=8.33(d,J=2.8Hz,1H),7.56-7.54(m,1H),7.52(d,J=3.2Hz,1H),7.30(dd,J=2.8Hz ,8.8Hz,1H),4.02-3.86(m,5H),3.34-3.04(m,2H),2.11-2.00(m,4H),1.45(s,9H). 19 F NMR (CDCl3400MHz)δ F =-129.12,-150.16. LCMSm / z[M+H-56] + 323.

[0513] Step 4: Synthesis of 5-fluoro-2-(4-fluoro-4-piperidinyl)-3-(1-methylpyrazol-4-yl)pyridine (C56): A mixture of tert-butyl 4-fluoro-4-[5-fluoro-3-(1-methylpyrazol-4-yl)-2-pyridinyl]piperidine-1-carboxylate (390 mg, 1.03 mmol) in EtOAc (1.00 mL) was mixed with HCl / EtOAc (4 M, 5.00 mL) and stirred at 20° C. for 1 hour. The mixture was concentrated to give 5-fluoro-2-(4-fluoro-4-piperidinyl)-3-(1-methylpyrazol-4-yl)pyridine, which was used in the next step without purification. LCMS m / z [M+H] + 279.

[0514] Step 5: Synthesis of tert-butyl 6-[4-fluoro-4-[5-fluoro-3-(1-methylpyrazol-4-yl)-2-pyridinyl]-1-piperidinyl]-2-azaspiro[3.4]octane-2-carboxylate (C57): A mixture of 5-fluoro-2-(4-fluoro-4-piperidinyl)-3-(1-methylpyrazol-4-yl)pyridine (324 mg, 1.03 mmol, HCl) in DCE (10.0 mL) was mixed with TEA (521 mg, 5.15 mmol) and tert-butyl 6-oxo-2-azaspiro[3.4]octane-2-carboxylate (255 mg, 1.13 mmol). The mixture was stirred at 25 ° C for 1 hour, then NaBH (OAc) was added (654.48 mg, 3.09 mmol) and AcOH (30.9 mg, 515 μmol) and the mixture was stirred at 25 ° C for 12 hours. The reaction mixture was mixed with saturated NaHCO aqueous solution (50 mL) and extracted with DCM (3 × 10 mL). The combined organic phase was washed with brine (20 mL), dried over anhydrous Na SO, filtered and concentrated to provide a residue. The residue was purified by flash silica gel chromatography (eluent was 0 to 5% MeOH / DCM) to obtain tert-butyl 6- [4- fluoro- 4- [5- fluoro- 3- (1- methylpyrazole -4- bases) -2- pyridyl] -1- piperidyl] -2- azaspiro [3.4] octane -2-carboxylate. 1 H NMR (CDCl3400MHz)δ H =8.33(d,J=2.8Hz,1H),7.57-7.49(m,2H),7.29(dd,J=2.8Hz,9.2Hz,1H),3.96(s,3H),3.85-3.68(m,4H),2 .84-2.72(m,2H),2.64-2.52(m,1H),2.40-2.06(m,7H),1.96-1.63(m,4H),1.55-1.50(m,1H),1.43(s,9H). LCMSm / z[M+H] + 488.

[0515] Step 6: Synthesis of 6-[4-fluoro-4-[5-fluoro-3-(1-methylpyrazol-4-yl)-2-pyridinyl]-1-piperidinyl]-2-azaspiro[3.4]octane (C58): A mixture of tert-butyl 6-[4-fluoro-4-[5-fluoro-3-(1-methylpyrazol-4-yl)-2-pyridinyl]-1-piperidinyl]-2-azaspiro[3.4]octane-2-carboxylate (200 mg, 410 μmol) in DCM (2.00 mL) was mixed with TFA (614 mg, 5.38 mmol) and stirred at 20°C for 12 hours. The mixture was concentrated to provide 6-[4-fluoro-4-[5-fluoro-3-(1-methylpyrazol-4-yl)-2-pyridinyl]-1-piperidinyl]-2-azaspiro[3.4]octane, which was used in the next step without further purification. LCMS m / z [M+H] + 388.

[0516] Step 7: Synthesis of ethyl 6-[4-fluoro-4-[5-fluoro-3-(1-methylpyrazol-4-yl)-2-pyridinyl]-1-piperidinyl]-2-azaspiro[3.4]octane-2-carboxylate (C59): A mixture of 6-[4-fluoro-4-[5-fluoro-3-(1-methylpyrazol-4-yl)-2-pyridinyl]-1-piperidinyl]-2-azaspiro[3.4]octane (205 mg, 409 μmol, TFA), TEA (124 mg, 1.23 mmol) in DCM (3.00 mL) at 0° C. was mixed with ethyl chloroformate (160 mg, 1.47 mmol) and added dropwise at 0° C. The mixture was then warmed to 20° C. and stirred at 20° C. for 4 hours. The reaction was slowly quenched with saturated aqueous NaHCO solution (10 mL) and then extracted with DCM (3×5 mL). The combined organic phases were washed with brine (10 mL), dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified by flash silica gel chromatography (eluent: 0 to 7% MeOH / DCM) to afford ethyl 6-[4-fluoro-4-[5-fluoro-3-(1-methylpyrazol-4-yl)-2-pyridinyl]-1-piperidinyl]-2-azaspiro[3.4]octane-2-carboxylate. LCMS m / z [M+H] + 460.

[0517] Step 8: Synthesis of (6R)-6-[4-fluoro-4-[5-fluoro-3-(1-methylpyrazol-4-yl)-2-pyridinyl]-1-piperidinyl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (245) and (6S)-6-[4-fluoro-4-[5-fluoro-3-(1-methylpyrazol-4-yl)-2-pyridinyl]-1-piperidinyl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (246): 6-[4-fluoro-4-[5-fluoro-3-(1-methylpyrazol-4-yl)-2-pyridinyl]-1-piperidinyl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester was purified by SFC (column: DAICEL CHIRALPAK AD (250mm×30mm, 10um); mobile phase: [CO2-EtOH (0.1% NH3H2O)]; B%: 20%, isocratic elution mode) to obtain (6R)-6-[4-fluoro-4-[5-fluoro-3-(1-methylpyrazol-4-yl)-2-pyridinyl]-1-piperidinyl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. Peak 1: 1 HNMR (CDCl3400MHz)δ H =8.32(d,J=2.4Hz,1H),7.57-7.50(m,2H),7.29(dd,J=2.8Hz,9.2Hz,1H),4.09(q,J=7.2Hz,2H),3.96(s,3H),3.91-3.72(m ,4H),2.92-2.73(m,2H),2.69-2.55(m,1H),2.47-2.01(m,7H),1.98-1.67(m,4H),1.64-1.48(m,1H),1.22(t,J=7.2Hz,3H). 19 F NMR (CDCl3400MHz)δ F =-75.75,-129.31. LCMSm / z[M+H] + 460. and (6S)-6-[4-fluoro-4-[5-fluoro-3-(1-methylpyrazol-4-yl)-2-pyridinyl]-1-piperidinyl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. Peak 2: 1 HNMR (CDCl3400MHz)δ H=8.32(d,J=2.8Hz,1H),7.59-7.48(m,2H),7.29(dd,J=2.8Hz,9.2Hz,1H),4.09(q,J=6.8Hz,2H),3.96(s,3H),3.91-3.72(m,4H),2.8 6-2.72(m,2H),2.66-2.54(m,1H),2.42-2.02(m,7H),1.96-1.72(m,1H),1.96-1.72(m,3H),1.60-1.47(m,1H),1.23(t,J=7.2Hz,3H). 19 F NMR (CDCl3400MHz)δ F =-75.76,-129.40. LCMSm / z[M+H] + 460.

[0518] Example 43: (6R)-6-[4-[5-fluoro-3-(4-fluorotetrahydropyran-4-yl)-2-pyridinyl]piperazin-1-yl]-2- Synthesis of Ethyl Azaspiro[3.4]octane-2-carboxylate (247)

[0519]

[0520] Step 1: Synthesis of (6R)-6-[4-[3-(3,6-dihydro-2H-pyran-4-yl)-5-fluoro-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (C60): (6R)-6-[4-(3-bromo-5-fluoro-2-pyridinyl)piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (230 mg, 521 μmol), 2-( A mixture of 3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (132 mg, 628 μmol), CsCO (340 mg, 1.04 mmol) and Pd(dppf)Cl (77.0 mg, 105 μmol) in 1,4-dioxane (8.00 mL) and HO (2.00 mL) was degassed and purged with N gas three times. The mixture was then stirred at 100° C. under N gas for 3 hours. The mixture was concentrated to provide a residue. The residue was purified by flash silica gel chromatography (0 to 10% MeOH / DCM as eluent) to afford (6R)-6-[4-[3-(3,6-dihydro-2H-pyran-4-yl)-5-fluoro-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester. 1 H NMR (DMSO-d 6400 MHz) δ H=8.11(d,J=3.2Hz,1H),7.39(dd,J=2.8Hz,8.8Hz,1H),6.03(br s,1H),4.21(br d,J=2.4Hz,2H),3.98(q,J=7.2Hz,2H),3.81-3.77(m,2H),3.77-3.63(m,4H),3.10(br s,4H),2.48-2.43(m,7H),2.08-2.00(m,1H),1.83-1.72(m,3H),1.66-1.59(m,1H),1.48-1.40(m,1H),1.14(t,J=7.2Hz,3H). LCMSm / z[M+H] + 445.

[0521] Step 2: Synthesis of (6R)-6-[4-[5-fluoro-3-(4-hydroxytetrahydropyran-4-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (C61): At 0°C, to (6R)-6-[4-[3-(3,6-dihydro-2H-pyran-4-yl)-5-fluoro-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester To a mixture of ethyl [3.4]octane-2-carboxylate (100 mg, 224 μmol), tri[(Z)-1-tert-butyl-4,4-dimethyl-3-oxo-pent-1-enyloxy]manganese (40.8 mg, 67.4 μmol) in i-PrOH (10.0 mL) was added phenylsilane (97.3 mg, 899 μmol) in DCE (4.00 mL). The mixture was then stirred at room temperature under O2 gas (15 psi) for 16 hours. The mixture was concentrated to provide a residue. The residue was purified by flash silica gel chromatography (eluent: 0 to 8% MeOH / DCM) to give ethyl (6R)-6-[4-[5-fluoro-3-(4-hydroxytetrahydropyran-4-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate. LCMS m / z [M+H] + 463.

[0522] Step 3: Synthesis of (6R)-6-[4-[5-fluoro-3-(4-fluorotetrahydropyran-4-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (247): A mixture of (6R)-6-[4-[5-fluoro-3-(4-hydroxytetrahydropyran-4-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylic acid ethyl ester (95.0 mg, 205 μmol) in DCM (10.0 mL) was degassed and purged with N gas three times. Then, DAST (170 mg, 1.06 mmol) in DCM (3.00 mL) was added dropwise to the mixture at 0° C., and the mixture was stirred under N gas at 0° C. for 2 hours. The pH was adjusted to 7-8 with saturated aqueous NaHCO₃, and the organic layer was washed with H₂O (2×10 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated to afford a residue. The residue was purified by preparative HPLC (column: Waters Xbridge, 150×25 mm×5 μm; mobile phase: [water (NH₃H₂O)-ACN]; gradient: 36%-66% B over 10 min) and lyophilized to afford ethyl (6R)-6-[4-[5-fluoro-3-(4-fluorotetrahydropyran-4-yl)-2-pyridinyl]piperazin-1-yl]-2-azaspiro[3.4]octane-2-carboxylate. 1 H NMR (CDCl3400MHz)δ H =8.24(d,J=2.8Hz,1H),7.62(dd,J=2.8Hz,9.6Hz,1H),4.10(q,J=7.2Hz,2H),3.96-3 .91(m,2H),3.91-3.87(m,2H),3.87-3.82(m,2H),3.80(s,2H),2.96(brs,4H),2.81- 2.70(m,1H),2.68-2.63(m,2H),2.62-2.54(m,2H),2.18-2.07(m,2H),1.98-1.89(m, 2H),1.88-1.79(m,2H),1.79-1.72(m,2H),1.71-1.51(m,2H),1.24(t,J=7.2Hz,3H). LCMSm / z[M+H] + 465.

[0523] Examples 37-234 were synthesized using the methods described above for Examples 1-36 and similar starting materials as shown in Table 1. Table 1 also shows the specific methods used, as well as the characterization data for these examples. Peak 1 or Peak 2 indicates the elution order on chiral HPLC / SFC.

[0524] Table 1

[0525]

[0526]

[0527]

[0528]

[0529]

[0530]

[0531]

[0532]

[0533]

[0534]

[0535]

[0536]

[0537]

[0538]

[0539]

[0540]

[0541]

[0542]

[0543]

[0544]

[0545]

[0546]

[0547]

[0548]

[0549]

[0550]

[0551]

[0552]

[0553]

[0554]

[0555]

[0556]

[0557]

[0558]

[0559]

[0560]

[0561]

[0562]

[0563]

[0564]

[0565]

[0566]

[0567]

[0568]

[0569]

[0570]

[0571]

[0572]

[0573]

[0574]

[0575]

[0576]

[0577]

[0578]

[0579]

[0580]

[0581] Biological assays

[0582] FLIPR assay

[0583] Fluorescence imaging plate reader (FLIPR) assay was performed using the intracellular calcium-sensitive dye Fluo 8, which exhibits increased fluorescence intensity when calcium is bound. Stimulation of the G-protein coupled receptor (GPCR) of the Gq coupling causes calcium to flow from the endoplasmic reticulum to the cytoplasm (Berridge, 1993), making this assay suitable for evaluating M1, M3, and M5 receptors. CHO-K1 cells overexpressing M1, M3, or M5 muscarinic acetylcholine receptors (mAChRs) were plated at 15,000 cells per well in a 30 μl volume and grown overnight at 37°C, 5% CO2. Fluo-8 solution was added to each well, 10 μl per well, and incubated at 37°C, 5% CO2 for 30 minutes. Compounds diluted in Hanks' balanced salt solution (HBSS) were transferred to a cell assay plate at 10 μl per well and then read on a FLIPR instrument. EC was calculated from the obtained data for each receptor subtype. 50 and E max number.

[0584] cAMP assay

[0585] M2 and M4 mACh receptors are G iCoupled receptors, causing a decrease in cAMP upon activation. An assay was developed to measure the potency and efficacy of compounds on M2 or M4 mAChRs using a CHO-K1 cell line overexpressing M2 or M4. Time-resolved fluorescence resonance energy transfer (TR-FRET) technology is used, in which a signal is generated as a result of energy transfer if the donor molecule is in close proximity to the acceptor molecule when the molecule has bound to the molecule of interest. The cAMP assay is a competitive binding assay in which cAMP produced by the cell competes with a labeled donor molecule for binding to an anti-cAMP receptor antibody. Due to the low basal level of cAMP in the CHO-K1 cell line, forskolin was used to increase cAMP levels to enable assessment of agonist activity at the mAChR of interest.

[0586] Table 2

[0587]

[0588]

[0589]

[0590]

[0591]

[0592]

[0593]

[0594]

[0595]

[0596]

[0597]

[0598]

[0599]

[0600] IA indicates IC 50 >10 μM; NT indicates not tested.

[0601] "+" means ≥100 nM; "++" means 20 to 99.9 nM; and "+++" means <20 nM;

[0602] "*" means 80% to 100%; "**" means 50% to 79.9%; and "***" means 19% to 49.9%; and "****" means <19%.

Claims

1. A compound having a structure of formula (I): or an N-oxide thereof, or a pharmaceutically acceptable salt of said compound or its N-oxide, wherein: A is a ring containing 1 or 2 nitrogen atoms and surrounded by 0, 1, 2 or 3 R A substituted 6-8 membered heterocycle; Each R A Independently C 1-3 Alkyl, halogen, =O, OH, C 1-3 Hydroxyalkyl or C 1-3 alkyl halide; Y is a bond, S, O, CH2, CHF, CF2 or C(OH)H; m is 1 or 2; n is 1 or 2; p is 1 or 2; R 1 H, halogen, CN, OH, NO2, -N(R 6 )(R 7 ), C 1-6 Alkyl, C 2-6 Heteroalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, -[O] 0-1 -C 3-6 Cycloalkyl, -[O] 0-1 -C 6-10 Aryl, -[O] 0-1 -4-8 membered heterocycle or -[O] 0-1 -5-10 membered heteroaryl, wherein the heterocycle and heteroaryl each contain 1, 2 or 3 ring heteroatoms selected from N, O and S, and when R 1 C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, -[O] 0-1 -C 3-6 Cycloalkyl, -[O] 0-1 -C 6-10 Aryl, -[O] 0-1 -4-8 membered heterocyclic ring, -[O] 0-1 -5-10 membered heteroaryl, -NH-C 3-6 Cycloalkyl, -NH-C 6-10 Aryl, -NH-4-8 membered heterocyclic ring, -NH-5-10 membered heteroaryl, -N(C 1-6 alkyl)-C 3-6 Cycloalkyl, -N(C 1-6 alkyl)-C 6-10 Aryl, -N(C 1-6 alkyl)-4-8 membered heterocyclic ring or -N(C 1-6 alkyl)-5-10 membered heteroaryl, R 1 0, 1, 2 or 3 independently selected from halogen, CN, OH, ═O, SO2 and C 1-3 Substitution of alkyl groups; R 2 H, halogen, CN, OH, –N(R 6 )(R 7 ), C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 2-6 Heteroalkyl, C 3-6 cycloalkyl or a 4-8 membered heterocycle containing 1, 2 or 3 ring heteroatoms selected from N, O and S; R 3 Halogen, C 0-6 Alkylene -CN, OH, -N(R 6 )(R 7 ), C 1-6 Alkyl, C 2-6 Heteroalkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, C 1-6 Alkylene-OC 1-6 Alkyl, C 1-6 Alkylene-NHC(O)C 1-6 Alkyl, C 1-6 Alkylene-C(O)NHC 1-6 Alkyl, C 0-6 Alkylene-Cyc, OC 0-6 Alkylene-Cyc, NH-Cyc, N(C 1-6 alkyl)-Cyc or C(O)Cyc; Cyc is C 3-12 Cycloalkyl, C 3-6 Heterocycloalkyl, C 5-12 Spiroalkyl, C 5-12 Heterospirocycloalkyl, C 6-10 aryl, 4-12 membered heterocyclic or 5-10 membered heteroaryl, said heterocyclic or heteroaryl containing 1, 2 or 3 ring heteroatoms independently selected from N, O and S, and Cyc is replaced by 0, 1, 2 or 3 R 3a Substituent substitution; Each R 3a independently selected from halogen, CN, OH, ═O, ═N(C 1-6 alkyl), SO2, C 1-6 Alkyl, C 2-10 Olefins, C 1-6 Hydroxyalkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 1-6 Alkylene-OC 1-6 Alkyl, C 0-6 Alkylene-C(O)C 1-6 Alkyl, C 0-6 Alkylene-NH2, C 0-6 Alkylene-NH(C 1-6 Alkyl), C 0-6 Alkylene-N(C 1-6 Alkyl)2, -SC 1-6 Alkyl, C 0-6 Alkylene-SO2C 1-6 Alkyl, C 0-6 Alkylene-C(O)NH2, C 0-6 Alkylene-C(O)NH(C 1-6 Alkyl), C 0-6 Alkylene-C(O)N(C 1-6 Alkyl)2, C 0-6 Alkylene-NHC(O)C 1-6 Alkyl, C 0-6 Alkylene-COOH, C 0-6 Alkylene-CO2C 1-6 Alkyl, C 0-6 Alkylene-C 3-6 Cycloalkyl and C containing 1, 2 or 3 heteroatoms selected from N, O and S 0-6 Alkylene-3-6 membered heterocycle; R 4 H, halogen, CN, OH, C 1-6 Alkyl, C 1-6 -haloalkyl or C 1-6 alkoxy; R 5 is -CO2-Z or its bioisostere; Each R 6 and R 7 Independently H, C 1-6 Alkyl, C(O)-C 1-6 Alkyl, spiro or bicyclic C 8-14 cycloalkyl, an 8-14 membered heterocycle containing 1, 2 or 3 ring heteroatoms selected from N, O and S, and when R 6 or R 7 When not H, it is replaced by 0, 1, 2 or 3 independently selected from halogen, CN, ═O, SO 2 , OH, C 0-6 Alkylene-NH2, C 0-6 Alkylene-NH(C 1-6 Alkyl), C 0-6 Alkylene-N(C 1-6 Alkyl)2, C 0-6 Alkylene-SO2C 1-6 Alkyl, C 1-6 Alkyl and C 1-6 The alkoxy substituent is substituted, or R 6 and R 7 together with the nitrogen to which they are attached, form a 4-10 membered heterocyclic ring containing 0-2 additional ring heteroatoms independently selected from N, O and S; and Z is C 1-7 Alkyl, C 1-7 Halogenated alkyl, C 3-6 Cycloalkyl or C 2-6 Alkyne, and Z is replaced by 0, 1, 2 or 3 C 1-6 Alkoxy or C 3-6 cycloalkyl substitution; The condition is that when R 1 and R 4 Each is H, m and p are each 1, and A is and (a)R 2 is H, Y is CH2, n is 1, and R 3 CH3, OCH3, or (b)R 2 is F, Y is CH2, n is 1, and R 3 for or (c)R 2 is H, Y is a bond, n is 1, and R 3 for or (d)R 2 is H, Y is a bond, n is 2, and R 3 for When R 5 Not CO2CH2CH3 or CO2CH(CH3)3.

2. The compound or salt according to claim 1, wherein A is and X is N, CH, C(OH) or CF.

3. The compound or salt according to claim 2, which has a structure of formula (Ia), or a pharmaceutically acceptable salt thereof:

4. The compound or salt according to any one of claims 1 to 3, wherein Y is CH2, CHF, CF2 or C(OH)H.

5. The compound or salt according to claim 4, wherein Y is CH2.

6. The compound or salt according to any one of claims 1 to 5, wherein m is 1.

7. The compound or salt according to any one of claims 1 to 6, wherein n is 1.

8. The compound or salt according to any one of claims 1 to 7, wherein p is 1.

9. The compound or salt according to claim 1, which has the structure of formula (Ib):

10. The compound or salt according to claim 9, which has the structure of formula (Ic):

11. The compound or salt according to claim 9, which has the structure of formula (Id):

12. The compound or salt according to claim 1, which has a structure of Formula (Ie), or a pharmaceutically acceptable salt thereof:

13. A compound or salt according to any one of claims 1 to 11, wherein R 5 is a CO2Z bioisostere and is selected from 14. A compound or salt according to any one of claims 1 to 12, wherein R 5 Selected from CO2C 1-7 alkyl, 15. The compound or salt according to claim 14, wherein R 5 It is CO2CH2CH3.

16. A compound or salt according to any one of claims 1 to 15, wherein R 1 H, halogen, CN, OH, -N(R 6 )(R 7 ), C 1-6 Alkyl or C 1-6 Alkoxy.

17. The compound or salt according to claim 16, wherein R 1 is H or halogen.

18. A compound or salt according to any one of claims 1 to 17, wherein R 2 H, halogen, CN, OH, -N(R 6 )(R 7 ), C 1-6 Alkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy or C 1-6 Halogenated alkoxy.

19. The compound or salt according to claim 18, wherein R 2 H, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl or C 1-6 Halogenated alkoxy.

20. The compound or salt according to claim 19, wherein R 2 is H or halogen.

21. A compound or salt according to any one of claims 1 to 20, wherein each R 6 and R 7 Independently H, C 1-6 Alkyl or C(O)-C 1-6 alkyl.

22. The compound or salt of claim 21, wherein each R 6 and R 7 are independently H or C 1-6 alkyl.

23. A compound or salt according to any one of claims 1 to 20, wherein at least one R 6 and R 7 Together with the nitrogen to which they are attached they form a 4-10 membered heterocyclic ring containing 0-2 additional ring heteroatoms independently selected from N and O.

24. A compound or salt according to any one of claims 1 to 23, wherein R 4 is H or halogen.

25. A compound or salt according to any one of claims 1 to 24, wherein R 1 、R 2 and R 4 At least one of them is a halogen.

26. A compound or salt according to any one of claims 1 to 25, wherein R 1 、R 2 and R 4 At least one of them is F.

27. A compound or salt according to any one of claims 1 to 26, wherein R 3 For -[O] 0-1 -C 3-6 Cycloalkyl, -[O] 0-1 -C 6-10 Aryl, -[O] 0-1 -4-8 membered heterocycle or -[O] 0-1 -5-10 membered heteroaryl, and R 3 0, 1, 2 or 3 R 3a replace.

28. The compound or salt according to claim 27, wherein R 3 C 3-6 cycloalkyl, 5-10 membered heteroaryl or 4-8 membered heterocycle, and R 3 0, 1, 2 or 3 R 3a replace.

29. A compound or salt according to any one of claims 1 to 26, wherein R 3 for and 0, 1, 2 or 3 R 3a replace.

30. The compound or salt of claim 29, wherein R 3 for and 0, 1, 2 or 3 R 3a replace.

31. A compound or salt according to any one of claims 1 to 30, wherein R 3 is unsubstituted.

32. A compound or salt according to any one of claims 1 to 30, wherein R 3 By 1 or 2 R 3a replace.

33. The compound or salt of claim 32, wherein R 3 By 1 R 3a replace.

34. A compound or salt according to any one of claims 1 to 30, 32 and 33, wherein at least one R 3a Halogen, CN, OH, =O, SO2, C 1-6 Alkyl, C 2-10 Olefins, C 1-6 Hydroxyalkyl, C 1-6 Halogenated alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkoxy, C 1-6 Alkylene-OC 1-6 Alkyl, N(C 1-6 Alkyl)2, C 1-6 Alkylene-N(C 1-6 Alkyl)2, -SC 1-6 Alkyl, NHC(O)C 1-6 Alkyl, C 1-6 Alkylene-NHC(O)C 1-6 an alkyl group or a C 0-6 Alkylene-3-6 membered heterocyclic ring.

35. The compound or salt of claim 34, wherein at least one R 3a For CH3, CH2CH3, CH(CH3)2, CH2CH(CH3)2, CF3, CH2CH2F, CH2CHF2, CH2OH, C(CH3)2OH, CH2OCH3, CH2CH2OCH3, CH2OCH2CH3, F, CN, =O, SO2, OH, OCH3, OCH2CH3, OCH(CH3)2, OCHF2, CH2OCH3、CH2OCF3、SCH3、N(CH3)2、NHCOCH3、CD3、 36. The compound or salt of claim 35, wherein at least one R 3a is CH3, CH2CH3, F, CN, OH, OCH3, CF3, CH2OH or OCHF2.

37. A compound as described in Table A or a pharmaceutically acceptable salt thereof.

38. The salt of claim 37, wherein the salt is selected from HCl, HCl.H2O, maleate, and maleate.H2O.

39. A pharmaceutical formulation comprising a therapeutically effective amount of a compound or salt according to any one of claims 1 to 38, and a pharmaceutically acceptable excipient.

40. A method for treating an M4-mediated (or M4-related) disease or disorder in a patient, the method comprising administering to the patient a therapeutically effective amount of a compound or salt according to any one of claims 1 to 38.

41. The method of claim 40, wherein the M4-mediated (or M4-related) disease or disorder is selected from Alzheimer's disease, schizophrenia or psychosis, pain, addiction, sleep disorders, cognitive disorders (e.g., mild cognitive impairment), Parkinson's disease, Parkinson's disease-levodopa-induced dyskinesia, Huntington's disease, movement disorders, dry mouth, pulmonary hypertension, chronic obstructive pulmonary disease (COPD), asthma, urinary incontinence, glaucoma, trisomy 21 (Down syndrome), cerebral amyloid angiopathy, Alzheimer's disease psychosis, dementia-related psychosis, bipolar disorder, hereditary cerebral hemorrhage with amyloidosis Dutch type (HCHWA-D), Creutzfeldt-Jakob disease, prion disorders, amyotrophic lateral sclerosis, progressive supranuclear palsy, head trauma, stroke, pancreatitis, inclusion body myositis, other peripheral amyloidoses, diabetes, autism, and atherosclerosis.

42. The method of claim 41, wherein the M4-mediated (or M4-related) disease or disorder is selected from Alzheimer's disease, schizophrenia, pain, addiction, Parkinson's disease, Parkinson's disease-levodopa-induced movement disorders, and sleep disorders.

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