Substituted spiro compound and application thereof

Abstract

Compounds of Formula (I) or stereoisomers or pharmaceutically acceptable salts thereof as TREM2 activators, processes for their preparation, pharmaceutical compositions containing the compounds of Formula (I) or stereoisomers or pharmaceutically acceptable salts thereof, and the Formula (I) Use of a compound or a stereoisomer thereof or a pharmaceutically acceptable salt thereof in the prevention or treatment of a TREM2 loss-of-function-related disease or disorder, including a neurodegenerative disease. # imgabs0 #

Description

[0001] This invention claims the priority of a prior application titled "Substituted Spiro Compounds and Their Applications" with patent application number CN202311838188.2, which was filed with the China National Intellectual Property Administration on December 27, 2023. The entire text of the above prior application is incorporated into this invention by reference. Technical Field

[0002] This invention relates to substituted spiro compounds, methods for their preparation, pharmaceutical compositions containing such compounds, and their use in preventing or treating diseases associated with loss of TREM2 function. Background Art

[0003] Microglia are innate immune cells present in the brain and are crucial for maintaining homeostatic conditions in the central nervous system. These cells, as macrophages in the brain, can sense microenvironmental changes through various receptors they express and alter their phenotypes to mediate responses to invading pathogens, proteotoxic stress, cell damage, etc. Microglia can rapidly proliferate upon stimulation and are characterized by exhibiting myeloid cell functions such as phagocytosis, cytokine / chemokine release, antigen presentation, and migration. Triggering receptor expressed on myeloid cells 2 (TREM2) is mainly expressed in microglia of the central nervous system (CNS) and is involved in the proliferation, survival, migration, and phagocytosis of microglia. In mice lacking functional TREM2 expression or expressing mutant receptors, the core observation is a weakened response of microglia to injury, such as oligodendrocyte demyelination, stroke-induced brain tissue damage, and proteotoxic inclusions in vivo.

[0004] In human genome-wide association studies, coding variants in the TREM2 locus are associated with late-onset Alzheimer's disease ("LOAD"), which links loss of TREM2 receptor function to increased disease risk. In the CNS, genetic variations in other genes selectively expressed by microglia (such as CD33, PLCg2, and MS4A4A / 6A) have reached genome-wide significance due to their association with LOAD risk. Collectively, these genetic findings highlight the importance of microglial innate immune function in LOAD. Additionally, an increase or elevation in soluble TREM2 ("sTREM2") in the cerebrospinal fluid (CSF) of human subjects is associated with the disease progression of LOAD and the emergence of pathological markers (including Tau protein phosphorylation). Natural history and human biology studies have shown that baseline sTREM2 levels in the CSF can stratify the rate of temporal lobe volume reduction and the rate of episodic memory decline in longitudinal monitoring cohorts.

[0005] In a rodent model with elevated TREM2 expression levels, cerebral amyloid pathology in 5XFAD transgenic mice showed reduced plaque volume and morphological changes. When TREM2 was overexpressed, changes in immunohistological markers of cerebral amyloid pathology were also accompanied by attenuation of dystrophic neurites. Therefore, pharmacological activation of TREM2 may serve as a potential target for treating or preventing neurological diseases, neurodegenerative diseases, and other diseases. SUMMARY OF THE INVENTION

[0006] The present invention relates to a compound of formula (I), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof,

[0007]

[0008] wherein,

[0009] X1 is selected from C(R a )2, NR b , O, S or S═O;

[0010] X3 is selected from a bond, C(R c )2 or C═O;

[0011] X7 is selected from a bond, O or C(R d )2;

[0012] X2, X4, X5, X6 are selected from C(R e )2;

[0013] X8 is selected from CR f or N;

[0014] Each R a , R b , R c , R d , R e , R f is independently selected from H, D, halogen, C1-C6 alkyl or C1-C6 alkoxy, and the C1-C6 alkyl or C1-C6 alkoxy is optionally substituted by D or halogen;

[0015] Ring A is selected from The a-terminal connection site is connected to Ring B;

[0016] Z1 is selected from CR 3 or N;

[0017] Ring B is selected from phenyl or a 5-6 membered heteroaryl;

[0018] R 1 is selected from C3-C 12 cycloalkyl, 4-12 membered heterocyclic group, C6-C 10An aryl or 5-10 membered heteroaryl, said C3-C 12 cycloalkyl, 4-12 membered heterocyclic group, C6-C 10 aryl or 5-10 membered heteroaryl is optionally substituted by R 1a ;

[0019] R 1a is selected from D, CN, halogen, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl or 4-7 membered heterocyclic group, said OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl or 4-7 membered heterocyclic group is optionally substituted by R 1b ;

[0020] R 1b is selected from D, CN, halogen, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl or 4-7 membered heterocyclic group, said OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl or 4-7 membered heterocyclic group is optionally substituted by R 1c ;

[0021] R 1c is selected from D, halogen or C1-C6 alkyl;

[0022] Each R 2 is independently selected from H, CN, halogen, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl or 4-7 membered heterocyclic group, said OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl or 4-7 membered heterocyclic group is optionally substituted by R 2a ;

[0023] R 2a is selected from D, halogen, C1-C6 alkyl or C1-C6 haloalkyl;

[0024] Each R 3 is independently selected from H, CN, halogen, C1-C6 alkyl or C1-C6 alkoxy, said C1-C6 alkyl or C1-C6 alkoxy is optionally substituted by R 3a ;

[0025] R 3a is selected from D or halogen;

[0026] m is selected from 0, 1, 2, 3, 4 or 5;

[0027] n is selected from 0, 1 or 2.

[0028] In some embodiments, Z1 is selected from N.

[0029] In some embodiments, ring A is selected from The a-terminal connection site is connected to ring B.

[0030] In some embodiments, Ra , R b , R c , R d , R e , R f is selected from H.

[0031] In some embodiments, is selected from

[0032] In some embodiments, R 1 is selected from a 4- to 7-membered heterocyclic group, a C3-C6 cycloalkyl group, or a 5- to 6-membered heteroaryl group, and the 4- to 7-membered heterocyclic group, C3-C6 cycloalkyl group, or 5- to 6-membered heteroaryl group is optionally substituted with R 1a .

[0033] In some embodiments, R 1 is selected from a pyridyl group, and the pyridyl group is optionally substituted with R 1a .

[0034] In some embodiments, R 1 is selected from the is optionally substituted with R 1a .

[0035] In some embodiments, R 1a is selected from a C1-C6 alkyl group or a C1-C6 alkoxy group.

[0036] In some embodiments, R 1a is selected from a methyl group or a methoxy group.

[0037] In some embodiments, R 1 is selected from

[0038] In some embodiments, R 2 is selected from H or a halogen.

[0039] In some embodiments, R 2 is selected from H, F, or Cl.

[0040] In some embodiments, R 3 is selected from H or a C1-C6 alkyl group.

[0041] In some embodiments, R 3 is selected from H or a methyl group.

[0042] In some embodiments, the compound of formula (I) of the present invention, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof is selected from the compound of formula (II), or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof,

[0043]

[0044]

[0045] wherein, R 1 、R 2 、R 3 、m and n are defined as above.

[0046] In some embodiments, the compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof of the present invention is selected from the compound of formula (III) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof,

[0047]

[0048] wherein, R 1 、R 2 and m are defined as above.

[0049] In some embodiments, the compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof of the present invention is selected from the following compounds or a stereoisomer thereof or a pharmaceutically acceptable salt thereof:

[0050]

[0051] On the other hand, the present invention provides a pharmaceutical composition comprising the compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof of the present invention and a pharmaceutically acceptable excipient.

[0052] On the other hand, the present invention provides a method for treating a disease associated with loss of TREM2 function in a mammal, comprising administering to a mammal in need of such treatment, preferably a human, a therapeutically effective amount of the compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0053] On the other hand, the present invention provides the use of the compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in the preparation of a medicament for preventing or treating a disease associated with loss of TREM2 function.

[0054] On the other hand, the present invention provides the use of the compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in preventing or treating a disease associated with loss of TREM2 function.

[0055] On the other hand, the present invention provides the compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for preventing or treating a disease associated with loss of TREM2 function.

[0056] In some embodiments, the diseases associated with loss of TREM2 function are selected from neurodegenerative diseases, preferably Alzheimer's disease.

[0057] Unless otherwise specified, the terms used in the present invention have the following meanings. The definitions of the groups and terms described in the present invention, including their exemplary definitions, illustrative definitions, preferred definitions, definitions recorded in tables, definitions of specific compounds in the examples, etc., can be combined and combined with each other arbitrarily. A particular term should not be considered indeterminate or unclear without a specific definition, but should be understood according to its ordinary meaning in the art. When a trade name appears in this article, it is intended to refer to the corresponding commodity or its active ingredient.

[0058] In this article, " " represents the linking site.

[0059] The graphical representation of racemic or enantiomerically pure compounds in this article is from Maehr, J. Chem. Ed. 1985, 62: 114 - 120. Unless otherwise specified, the absolute configuration of a stereocenter is represented by a wedge bond and a dashed wedge bond ( and ), and the relative configuration of a stereocenter (such as the cis - trans configuration of an alicyclic compound) is represented by a solid black bond and a dashed bond ( and ).

[0060] The term "tautomer" refers to functional group isomers resulting from the rapid movement of an atom in a molecule between two positions. The compounds of the present invention may exhibit tautomerism. Tautomeric compounds can exist in two or more interconvertible forms. Tautomers generally exist in an equilibrium form. When attempting to isolate a single tautomer, a mixture is usually produced, and its physicochemical properties are consistent with those of a mixture of compounds. The position of the equilibrium depends on the chemical characteristics within the molecule. For example, in many aliphatic aldehydes and ketones such as acetaldehyde, the keto form predominates; while in phenols, the enol form predominates. The present invention encompasses all tautomeric forms of the compounds.

[0061] The term "stereoisomer" refers to isomers resulting from different spatial arrangements of atoms in a molecule, including cis - trans isomers, enantiomers, and diastereomers.

[0062] The compounds of the present invention may have asymmetric atoms such as carbon, sulfur, nitrogen, phosphorus atoms or asymmetric double bonds, and thus the compounds of the present invention may exist in specific geometric or stereoisomeric forms. Specific geometric or stereoisomeric forms may be cis- and trans-isomers, E- and Z-geometric isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereoisomers, (D)-isomers, (L)-isomers, and their racemic mixtures or other mixtures, such as enantiomer- or diastereomer-enriched mixtures. All of the above isomers and their mixtures are within the definition scope of the compounds of the present invention. Additional asymmetric carbon, sulfur, nitrogen or phosphorus atoms may exist in substituents such as alkyl groups, and these isomers and their mixtures involved in all substituents are also included within the definition scope of the compounds of the present invention. The compounds of the present invention containing asymmetric atoms may be isolated in optically pure form or in racemic form. The optically pure form may be resolved from the racemic mixture or synthesized by using chiral starting materials or chiral reagents.

[0063] The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, provided that the valence of the particular atom is normal and the resulting compound is stable. When the substituent is oxo (i.e., =O), it means that two hydrogen atoms are replaced, and oxo does not occur on an aromatic group.

[0064] The term "optionally" or "optionally" means that the subsequent described event or situation may or may not occur, and this description includes the occurrence and non-occurrence of the described event or situation. For example, when ethyl is "optionally" substituted by a halogen, it means that ethyl can be unsubstituted (CH2CH3), monosubstituted (CH2CH2F, CH2CH2Cl, etc.), polysubstituted (CHFCH2F, CH2CHF2, CHFCH2Cl, CH2CHCl2, etc.) or fully substituted (CF2CF3, CF2CCl3, CCl2CCl3, etc.). Those skilled in the art will understand that for any group containing one or more substituents, no substitution or substitution pattern that is spatially impossible to exist and / or cannot be synthesized will be introduced.

[0065] When any variable (e.g., R a , R b ) appears more than once in the composition or structure of a compound, its definition in each case is independent. For example, if a group is substituted by two R b , then each R b has an independent option.

[0066] When the number of a linking group is 0, such as -(CH2)0-, it means that the linking group is a bond.

[0067] When one of the variables is selected from a chemical bond or is absent, it means that the two groups it connects are directly linked. For example, in A-L-Z, when L represents a bond, it means the structure is actually A-Z.

[0068] When a bond of a substituent cross-links to two atoms on a ring, this substituent can be bonded to any atom on this ring. For example, the structural unit means that R 2 can be substituted at any position on the benzene ring.

[0069] C m -C n in this article refers to a hydrocarbon group having an integer number of carbon atoms in the range of m - n. For example, "C1-C 10 " means that this group can have 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, 8 carbon atoms, 9 carbon atoms, or 10 carbon atoms.

[0070] The term "alkyl" refers to a hydrocarbon group with the general formula C n H 2n+1 , and this alkyl can be straight-chain or branched-chain. The term "C1-C6 alkyl" can be understood to represent a straight-chain or branched-chain saturated monovalent hydrocarbon group having 1, 2, 3, 4, 5, or 6 carbon atoms. Specific examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, neopentyl, hexyl, 2-methylpentyl, etc. The term "C1-C3 alkyl" can be understood to represent a straight-chain or branched-chain saturated monovalent hydrocarbon group having 1 to 3 carbon atoms. The said "C1-C6 alkyl" can further contain "C1-C3 alkyl".

[0071] The term "alkoxy" refers to a monovalent group formed by removing the hydrogen atom on the hydroxyl group of a straight-chain or branched-chain alcohol, and can be understood as "alkyloxy" or "alkyl - O -". The term "C1-C6 alkoxy" can be understood as "C1-C6 alkyloxy" or "C1-C6 alkyl - O -". The said "C1-C6 alkoxy" can further contain "C1-C3 alkoxy".

[0072] The term "halo" or "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0073] The term "C1-C6 haloalkyl" includes monohalogenated C1-C6 alkyl or polyhalogenated C1-C6 alkyl.

[0074] The term "cycloalkyl" refers to a completely saturated carbocyclic ring existing in the form of a monocyclic, fused-ring, bridged-ring, or spiro-ring, etc. Unless otherwise indicated, this carbocyclic ring is usually a 3- to 12-membered ring. The term "C3-C12 "Cycloalkyl" should be understood to mean a saturated monovalent monocyclic, fused-ring, spiro or bridged ring having 3 to 12 carbon atoms. Specific examples of the cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, norbornyl (bicyclo[2.2.1]heptyl), bicyclo[2.2.2]octyl, adamantyl, spiro[4.5]decyl, etc. The term "C3-C 12 cycloalkyl" may include "C3-C6 cycloalkyl", and the term "C3-C6 cycloalkyl" can be understood to mean a saturated monovalent monocyclic or bicyclic hydrocarbon ring having 3 to 6 carbon atoms. Specific examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, etc.

[0075] The term "heterocyclic group" refers to a monocyclic, fused-ring, spiro or bridged ring group that is fully saturated or partially saturated (not heteroaromatic with aromaticity as a whole), and contains 1-5 heteroatoms or heteroatom groups (i.e., atomic groups containing heteroatoms) among its ring atoms. The "heteroatom or heteroatom group" includes, but is not limited to, nitrogen atom (N), oxygen atom (O), sulfur atom (S), phosphorus atom (P), boron atom (B), -S(=O)2-, -S(=O)-, and optionally substituted -NH-, -S(=O)(=NH)-, -C(=O)NH-, -C(=NH)-, -S(=O)2NH-, S(=O)NH- or -NHC(=O)NH-, etc. The term "4-12 membered heterocyclic group" refers to a heterocyclic group having 4, 5, 6, 7, 8, 9, 10, 11 or 12 ring atoms, and containing 1-5 heteroatoms or heteroatom groups independently selected from the above-mentioned heteroatoms or heteroatom groups among its ring atoms. The "4-12 membered heterocyclic group" includes the "4-7 membered heterocyclic group". Among them, specific examples of the 4-membered heterocyclic group include, but are not limited to, azetidinyl or oxetanyl; specific examples of the 5-membered heterocyclic group include, but are not limited to, tetrahydrofuranyl, dioxolanyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrroline, 4,5-dihydrooxazolyl or 2,5-dihydro-1H-pyrrolyl; specific examples of the 6-membered heterocyclic group include, but are not limited to, tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, trithianyl, tetrahydropyridyl or 4H-[1,3,4]thiadiazinyl; specific examples of the 7-membered heterocyclic group include, but are not limited to, diazepanyl. The heterocyclic group can also be a bicyclic group. Among them, specific examples of the 5,5 bicyclic group include, but are not limited to, hexahydrocyclopenta[c]pyrrol-2(1H)-yl; specific examples of the 5,6 bicyclic group include, but are not limited to, hexahydropyrrolo[1,2-a]pyrazin-2(1H)-yl, 5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazinyl or 5,6,7,8-tetrahydroimidazo[1,5-a]pyrazinyl. Optionally, the heterocyclic group can be a benzo-fused ring group of the above 4-7 membered heterocyclic group, and specific examples include, but are not limited to, dihydroisoquinolinyl, etc. The "4-12 membered heterocyclic group" can include ranges such as "5-12 membered heterocyclic group", "4-7 membered heterocyclic group", "5-6 membered heterocyclic group", "6-8 membered heterocyclic group", "4-12 membered heterocycloalkyl group", "5-12 membered heterocycloalkyl group", "4-7 membered heterocycloalkyl group", "5-6 membered heterocycloalkyl group", "6-8 membered heterocycloalkyl group", etc. The "4-7 membered heterocyclic group" can further include ranges such as "4-6 membered heterocyclic group", "5-6 membered heterocyclic group", "4-7 membered heterocycloalkyl group", "4-6 membered heterocycloalkyl group", "5-6 membered heterocycloalkyl group", etc. Although some bicyclic heterocyclic groups in the present invention partially contain a benzene ring or a heteroaromatic ring, the heterocyclic group as a whole is still non-aromatic.

[0076] The term "aryl" refers to an aromatic ring group of a fully carbonaceous monocyclic or fused polycyclic having a conjugated π electron system. The term "C6-C 10 aryl" should be understood as a monovalent aromatic or partially aromatic fully carbonaceous monocyclic or bicyclic group having 6 to 10 carbon atoms. In particular, a ring having 6 carbon atoms ("C6 aryl"), such as phenyl; or a ring having 9 carbon atoms ("C9 aryl"), such as indanyl or indenyl; or a ring having 10 carbon atoms ("C 10 aryl"), such as tetrahydronaphthyl, dihydronaphthyl or naphthyl.

[0077] The term "heteroaryl" refers to a monocyclic or fused polycyclic system having aromaticity, wherein the ring atoms contain at least one ring atom selected from N, O, S, and the remaining ring atoms are C aromatic ring groups. The term "5-10 membered heteroaryl" should be understood to include such a monovalent monocyclic or bicyclic aromatic ring system: having 5, 6, 7, 8, 9 or 10 ring atoms, especially 5 or 6 or 9 or 10 ring atoms, and containing 1-5, preferably 1-3 heteroatoms independently selected from N, O and S. In particular, heteroaryl is selected from thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl or thiadiazolyl, etc. and their benzo derivatives, such as benzofuryl, benzothienyl, benzothiazolyl, benzoxazolyl, benzisoxazolyl, benzimidazolyl, benzotriazolyl, indazolyl, indolyl or isoindolyl, etc.; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl or triazinyl, etc. and their benzo derivatives, such as quinolinyl, quinazolinyl or isoquinolinyl, etc.; or azocinyl, indolizinyl, purinyl, etc. and their benzo derivatives; or cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, pteridinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl or phenoxazinyl, etc. The term "5-6 membered heteroaryl" refers to an aromatic ring system having 5 or 6 ring atoms, and containing 1-3, preferably 1-2 heteroatoms independently selected from N, O and S.

[0078] The term "therapeutically effective amount" means the amount of the compound of the present invention that (i) treats or prevents a particular disease, condition or disorder, (ii) alleviates, ameliorates or eliminates one or more symptoms of a particular disease, condition or disorder, or (iii) prevents or delays the onset of one or more symptoms of a particular disease, condition or disorder described herein. The amount of the compound of the present invention constituting a "therapeutically effective amount" varies depending on the compound, the disease state and its severity, the mode of administration, and the age of the mammal to be treated, but can be routinely determined by those skilled in the art based on their own knowledge and the present disclosure.

[0079] The term "pharmaceutically acceptable" refers to compounds, materials, compositions, and / or dosage forms that, within the scope of sound medical judgment, are suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0080] The term "pharmaceutically acceptable salt" refers to salts of pharmaceutically acceptable acids or bases, including salts formed by the compound with inorganic acids or organic acids, and salts formed by the compound with inorganic bases or organic bases.

[0081] The term "pharmaceutical composition" refers to a mixture of one or more compounds of the present invention or their salts and pharmaceutically acceptable excipients. The purpose of the pharmaceutical composition is to facilitate the administration of the compounds of the present invention to an organism.

[0082] The term "pharmaceutically acceptable excipient" refers to those excipients that do not cause significant irritation to the organism and do not impair the biological activity and properties of the active compound. Suitable excipients are well known to those skilled in the art, such as carbohydrates, waxes, water-soluble and / or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, etc.

[0083] The words "comprise" or "comprising" and their English variants such as "comprises" or "comprising" shall be understood in an open, non-exclusive sense, i.e., "including but not limited to".

[0084] The present invention also includes isotopically labeled compounds of the present invention that are the same as those described herein, but in which one or more atoms are replaced by atoms having an atomic weight or mass number different from the atomic weight or mass number commonly found in nature. Examples of isotopes that can be incorporated into the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine, and chlorine, such as 2 H, 3 H, 11 C, 13 C, 14 C, 13 N, 15 N, 15 O, 17 O, 18 O, 31 P, 32 P, 35 S, 18 F, 123 I, 125 I, and 36 Cl, etc.

[0085] Certain isotopically labeled compounds of the present invention (e.g., those labeled with3 H and 14 C labeling) can be used in the analysis of the tissue distribution of compounds and / or substrates. Tritiation (i.e., 3 H) and carbon-14 (i.e., 14 C) isotopes are particularly preferred due to their ease of preparation and detectability. Positron-emitting isotopes, such as 15 O, 13 N, 11 C, and 18 F can be used in positron emission tomography (PET) studies to determine substrate occupancy. Isotopically labeled compounds of the present invention can generally be prepared by substituting an isotopically unlabeled reagent with an isotopically labeled reagent by procedures similar to those described in the protocols and / or examples disclosed below.

[0086] The pharmaceutical compositions of the present invention can be prepared by combining the compounds of the present invention with suitable pharmaceutically acceptable excipients, and can be formulated into solid, semi-solid, liquid or gaseous preparations, such as tablets, pills, capsules, powders, granules, ointments, emulsions, suspensions, suppositories, injections, inhalants, gels, microspheres, and aerosols, etc.

[0087] Typical routes of administration of the compounds of the present invention or their pharmaceutically acceptable salts or their pharmaceutical compositions include, but are not limited to, oral, rectal, topical, inhaled, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, intravenous administration.

[0088] The pharmaceutical compositions of the present invention can be manufactured by methods well known in the art, such as conventional mixing methods, dissolution methods, granulation methods, emulsification methods, freeze-drying methods, etc.

[0089] In some embodiments, the pharmaceutical composition is in oral form. For oral administration, the pharmaceutical composition can be formulated by mixing the active compound with pharmaceutically acceptable excipients well known in the art. These excipients enable the compounds of the present invention to be formulated into tablets, pills, lozenges, dragees, capsules, liquids, gels, syrups, suspensions, etc. for oral administration to a patient.

[0090] Solid oral compositions can be prepared by conventional mixing, filling or tableting methods. For example, they can be obtained by mixing the active compound with solid excipients, optionally milling the resulting mixture, adding other suitable excipients if necessary, and then processing the mixture into granules to obtain the core of a tablet or dragee. Suitable excipients include, but are not limited to, binders, diluents, disintegrants, lubricants, glidants or flavoring agents, etc.

[0091] The pharmaceutical composition is also suitable for parenteral administration, such as sterile solutions, suspensions or lyophilized products in suitable unit dosage forms.

[0092] In all methods of administering the compounds of general formula I described herein, the daily dosage is from 0.01 mg / kg to 200 mg / kg body weight, preferably from 0.05 mg / kg to 50 mg / kg body weight, more preferably from 0.1 mg / kg to 30 mg / kg body weight, in the form of single or divided doses.

[0093] The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, the embodiments formed by their combination with other chemical synthesis methods, and equivalent replacement methods well known to those skilled in the art. Preferred embodiments include, but are not limited to, the examples of the present invention.

[0094] The chemical reactions of the specific embodiments of the present invention are completed in a suitable solvent, and the solvent must be suitable for the chemical changes of the present invention and the reagents and materials required therefor. In order to obtain the compounds of the present invention, it is sometimes necessary for those skilled in the art to modify or select the synthesis steps or reaction processes on the basis of the existing embodiments.

[0095] The following abbreviations are used in the present invention:

[0096] HATU: O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate; DCM: dichloromethane; PE: petroleum ether; DMF: N,N-dimethylformamide; DMSO: dimethyl sulfoxide; EA: ethyl acetate; BOC: tert-butylcarbonyl; AcOH: acetic acid; THF: tetrahydrofuran; TFA: trifluoroacetic acid; DIEA or DIPEA: diisopropylethylamine; Pd(dppf)Cl2: 1,1-bis(diphenylphosphino)ferrocene palladium chloride Specific Embodiments

[0097] The invention will be described in detail below by way of examples, which does not mean any adverse limitation to the present invention. The present invention has been described in detail herein, and specific embodiments thereof have also been disclosed. It will be obvious to those skilled in the art to make various changes and improvements to the specific embodiments of the present invention without departing from the spirit and scope of the present invention. All reagents used in the present invention are commercially available and can be used without further purification.

[0098] Unless otherwise stated, the ratios indicated for mixed solvents are volume mixing ratios. Unless otherwise stated, % means wt%.

[0099] The compounds are named by hand or software, and commercially available compounds use the supplier catalog names.

[0100] The structure of the compound was determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). The unit of NMR shift is 10 -6 (ppm). The solvents for NMR determination were deuterated dimethyl sulfoxide, deuterated chloroform, deuterated methanol, etc., and the internal standard was tetramethylsilane (TMS);

[0101] The eluent below can be formed into a mixed eluent by two or more solvents, and the ratio is the volume ratio of each solvent. For example, "0-10% methanol / dichloromethane" means that during the gradient elution process, the volume ratio of methanol to dichloromethane in the mixed eluent is 0:100-10:100.

[0102] Example 1: Preparation of 8-(4-chloro-2-fluorophenyl)-2,3-dimethyl-6-(2-(2-methylpyridin-4-yl)-5-oxa-2,8-diazaspiro[3.5]nonan-8-yl)pyrimido[5,4-d]pyrimidin-4(3H)-one (Compound 1)

[0103]

[0104] Step 1: Preparation of ethyl 5-amino-2-chloro-6-(4-chloro-2-fluorophenyl)pyrimidine-4-carboxylate (1C)

[0105] Compound 1A (2 g, 8.47 mmol), 1B (1.47 g, 8.47 mmol), Pd(dppf)Cl2 (1.23 g, 1.68 mmol) and K3PO4 (5.4 g, 25.4 mmol) were mixed, and a mixed solution of 1,4-dioxane and water (55 mL, 1,4-dioxane: water = 10:1) was added under nitrogen protection, and the reaction was carried out at 55 °C. After the reaction was detected to be complete by TLC, it was cooled to room temperature, quenched with water, extracted with EA, the organic phase was dried with anhydrous sodium sulfate, filtered, the organic phase was concentrated, and the crude product was separated by silica gel column chromatography (PE:EA = 3:1) to obtain the title compound 1C (1.6 g).

[0106] 1 H NMR (300 MHz, DMSO-d6) δ 7.71-7.44 (m, 3H), 6.70 (s, 2H), 4.37 (q, J = 7.1 Hz, 2H), 1.34 (t, J = 7.1 Hz, 3H).

[0107] Step 2: Preparation of 5-amino-2-chloro-6-(4-chloro-2-fluorophenyl)pyrimidine-4-carboxylic acid (1D)

[0108] To a solution of Compound 1C (1.8 g, 5.47 mmol) in THF (30 mL) was added a solution of LiOH (1.15 g, 27.4 mmol) in water (20 mL), and the mixture was stirred at room temperature. After the reaction was complete as detected by TLC, the reaction was quenched with water and aqueous HCl solution (2 M, 30 mL), extracted with DCM, the organic phase was dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated to obtain the crude product of the title compound 1D (1.6 g), which was directly used in the next step without purification.

[0109] Step 3: Preparation of 5-Amino-2-chloro-6-(4-chloro-2-fluorophenyl)-N-methylpyrimidine-4-carboxamide (1E)

[0110] Compound 1D (1.27 g, 4.2 mmol), methylamine hydrochloride (428 mg, 6.3 mmol) and HATU (2.4 g, 6.3 mmol) were mixed, DMF (30 mL) was added under nitrogen protection, and the mixture was stirred at 20 °C, then DIEA (1.63 g, 12.7 mmol) was added, and the reaction was carried out at 50 °C. After the reaction was complete as detected by TLC, the reaction was quenched with water, extracted with EA, the organic phase was dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated. The crude product was separated by silica gel column chromatography (PE:EA = 3:1) to obtain the title compound 1E (1.24 g).

[0111] 1 1H NMR (300 MHz, DMSO-d6) δ 8.87 (d, J = 5.1 Hz, 1H), 7.74 - 7.44 (m, 3H), 6.92 (s, 2H), 2.79 (d, J = 4.8 Hz, 3H).

[0112] Step 4: Preparation of 6-Chloro-8-(4-chloro-2-fluorophenyl)-2,3-dimethylpyrimido[5,4-d]pyrimidin-4(3H)-one (1F)

[0113] Compound 1E (1.24 g, 3.95 mmol), 1,1,1-triethoxyethane (5 mL) and AcOH (5 mL) were mixed and stirred at 100 °C. After the reaction was complete as detected by TLC, the mixture was cooled to room temperature, quenched with water, extracted with EA, the organic phase was dried over anhydrous sodium sulfate, filtered, and the organic phase was concentrated. The crude product was separated by silica gel column chromatography (PE:EA = 1:1) to obtain the title compound 1F (900 mg).

[0114] 1 1H NMR (300 MHz, CDCl3) δ 7.66 (d, J = 8.3 Hz, 1H), 7.32 (d, J = 7.3 Hz, 1H), 7.24 (d, J = 1.7 Hz, 1H), 3.69 (s, 3H), 2.61 (s, 3H).

[0115] Step 5: Preparation of tert-butyl 8-(4-(4-chloro-2-fluorophenyl)-6,7-dimethyl-8-oxo-7,8-dihydropyrimido[5,4-d]pyrimidin-2-yl)-5-oxa-2,8-diazaspiro[3.5]nonane-2-carboxylate (1H)

[0116] Compound 1F (120 mg, 0.35 mmol), 1G (121 mg, 0.53 mmol), and DIPEA (137 mg, 1.06 mmol) were dissolved in DMSO and reacted at 80 °C. After the reaction was completed as detected by TLC, it was cooled to room temperature, quenched with water, extracted with EA, the organic phase was dried over anhydrous sodium sulfate, filtered, the organic phase was concentrated, and the crude product was separated by silica gel column chromatography (PE:EA = 1:1) to obtain the title compound 1H (110 mg).

[0117] 1 1H NMR (300 MHz, DMSO-d6) δ 7.68 (t, J = 8.0 Hz, 1H), 7.62 (dd, J = 9.8, 2.0 Hz, 1H), 7.47 (dd, J = 8.3, 2.1 Hz, 1H), 3.96 (s, 2H), 3.74 (m, 8H), 3.52 (s, 3H), 2.45 (s, 3H), 1.37 (s, 9H).

[0118] Step 6: Preparation of trifluoroacetate of 8-(4-chloro-2-fluorophenyl)-2,3-dimethyl-6-(5-oxa-2,8-diazaspiro[3.5]nonan-8-yl)pyrimido[5,4-d]pyrimidin-4(3H)-one (1I)

[0119] Compound 1H (110 mg, 0.21 mmol) was dissolved in DCM (6 mL), TFA (1 mL) was added, and the mixture was stirred at room temperature. After the reaction was completed as detected by TLC, the reaction solution was concentrated to obtain the crude trifluoroacetate of the title compound 1I, which was directly used for the next step without purification.

[0120] Step 7: Preparation of 8-(4-chloro-2-fluorophenyl)-2,3-dimethyl-6-(2-(2-methylpyridin-4-yl)-5-oxa-2,8-diazaspiro[3.5]nonan-8-yl)pyrimido[5,4-d]pyrimidin-4(3H)-one (Compound 1)

[0121] The trifluoroacetate of compound 1I (110 mg, 0.2 mmol), 1J (34 mg, 0.3 mmol), and Cs2CO3 (199 mg, 0.6 mmol) were dissolved in DMSO (5 mL) and reacted at 110 °C. After the reaction was completed as detected by TLC, it was cooled to room temperature, quenched with water, extracted with EA, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, the organic phase was concentrated, and separated by silica gel column chromatography (PE:EA = 3:1) to obtain the title compound 1 (52 mg).

[0122] 1 1H NMR (300 MHz, CDCl3) δ 8.19–8.08 (m, 1H), 7.61 (t, J = 7.9 Hz, 1H), 7.31 (d, J = 2.0 Hz, 1H), 7.24 (dd, J = 9.6, 1.9 Hz, 1H), 6.17 (d, J = 5.5 Hz, 2H), 4.18 (s, 2H), 4.06–3.80 (m, 8H), 3.64 (s, 3H), 2.54 (s, 3H), 2.48 (s, 3H).

[0123] MS m / z (ESI): = 522.20 [M+H] + .

[0124] Example 2: Preparation of 8-(4-chloro-2-fluorophenyl)-2,3-dimethyl-6-(2-(2-methoxypyridin-4-yl)-5-oxa-2,8-diazaspiro[3.5]nonan-8-yl)pyrimido[5,4-d]pyrimidin-4(3H)-one (Compound 2)

[0125]

[0126] The trifluoroacetate of compound 1I (110 mg, 0.2 mmol), 2A (38 mg, 0.3 mmol), and DIPEA (199 mg, 0.6 mmol) were dissolved in DMSO (5 mL) and reacted at 80 °C. After the reaction was completed as detected by TLC, it was cooled to room temperature, quenched with water, extracted with EA, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, the organic phase was concentrated, and separated by silica gel column chromatography (EA = 100%) to obtain the title compound 2 (69 mg).

[0127] 11H NMR (300 MHz, CDCl3) δ 7.90 (dd, J = 5.8, 2.7 Hz, 1H), 7.65 (td, J = 7.9, 2.7 Hz, 1H), 7.36–7.26 (m, 2H), 6.06 (dt, J = 5.6, 2.4 Hz, 1H), 5.71 (d, J = 2.3 Hz, 1H), 4.19 (d, J = 3.0 Hz, 2H), 4.07–3.94 (m, 4H), 3.93 (s, 3H), 3.86 (t, J = 4.8 Hz, 4H), 3.67 (s, 3H), 2.57 (s, 3H).

[0128] Test Examples of Biological Activity and Related Properties

[0129] Test Example 1: Detection of Phosphorylation of Spleen Tyrosine Kinase Syk (pSyk) in Downstream Cells

[0130] Experimental Materials

[0131]

[0132] Experimental Methods

[0133] HEK-293T cells overexpressing hTREM2 / DAP12 were routinely cultured in DMEM medium (containing 10% FBS and 7 μg / mL Puromycin). The cells were digested with Trypsin-EDTA, centrifuged, the supernatant was removed, and the cells were resuspended with the medium; the density of the overexpressing cells was adjusted to 50,000 cells / mL, dispensed into 96-well plates, 100 μL per well, and incubated overnight in an incubator at 37 °C and 5% CO2.

[0134] Aspirate the supernatant, add DMEM without serum (containing 7 μg / mL Puromycin) medium, and culture the cells in a 37°C, 5% CO2 incubator for 2 - 3 h. Add the test compounds (at concentrations of 30 pM, 1 nM, 30 nM, 1 μM, and 30 μM) prepared with serum-free DMEM (containing 7 μg / mL Puromycin) medium to the cells, and then incubate them in a 37°C, 5% CO2 incubator for 15 min. Remove the supernatant, and according to the AlphaLISA SureFire Ultra p-SYK kit instructions, prepare and add the corresponding reagents in sequence: Add 50 μL of 1x lysis buffer to each well, incubate at room temperature for 10 min, and centrifuge at 400 rpm for 10 min. Transfer 10 μL of cell lysate to a new 384-well plate, add 5 μL of acceptor mix to each well, mix by shaking, and incubate in the dark for 1 h. Add 5 μL of donor mix to each well, mix by shaking, and incubate in the dark overnight. Centrifuge at 1,000 rpm for 1 min, and use a microplate reader (brand: SpectraMax) to read the absorbance of each well using the αLISA program (E x : 680 nm, E m : 570 nm).

[0135] The tests were performed in triplicate. Using the sample readings of the medium control group (without compounds) as the base number 1, calculate the relative readings (pSyk signal window, relative to medium only) at each concentration of the test compound. Plot the relative readings against the concentration, and use GraphPad Prism software to perform four-parameter non-linear fitting to calculate the EC 50 .

[0136] Test results:

[0137] Table 1: Effects of test compounds on SYK phosphorylation in hTREM2 / DAP12-overexpressing HEK-293T cells

[0138]

Claims

1. A compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, in, X1 is selected from C(R a )2. NR b , O, S or S=O; X3 is selected from a bond, C(R c )2 or C=O; X7 is selected from a bond, O or C(R d )2; X2, X4, X5, X6 are selected from C(R e )2; X8 from CR f or N; Each R a , R b , R c , R d , R e , R f is independently selected from H, D, halogen, C1-C6 alkyl or C1-C6 alkoxy, wherein the C1-C6 alkyl or C1-C6 alkoxy is optionally substituted by D or halogen; Ring A is selected from The a-terminal attachment site is connected to loop B; Z1 is selected from CR 3 or N; Ring B is selected from phenyl or 5-6 membered heteroaryl; R 1 Selected from C3-C 12 Cycloalkyl, 4-12 membered heterocyclic group, C6-C 10 Aryl or 5-10 membered heteroaryl, the C3-C 12 Cycloalkyl, 4-12 membered heterocyclic group, C6-C 10 Aryl or 5-10 membered heteroaryl is optionally replaced by R 1a replace; R 1a is selected from D, CN, halogen, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl or 4-7 membered heterocyclic group, wherein the OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl or 4-7 membered heterocyclic group is optionally replaced by R 1b replace; R 1b is selected from D, CN, halogen, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl or 4-7 membered heterocyclic group, wherein the OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl or 4-7 membered heterocyclic group is optionally replaced by R 1c replace; R 1c Selected from D, halogen or C1-C6 alkyl; Every R 2 independently selected from H, CN, halogen, OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl or 4-7 membered heterocyclyl, wherein the OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl or 4-7 membered heterocyclyl is optionally replaced by R 2a replace; R 2a is selected from D, halogen, C1-C6 alkyl or C1-C6 haloalkyl; Each R 3 independently selected from H, CN, halogen, C1-C6 alkyl or C1-C6 alkoxy, wherein the C1-C6 alkyl or C1-C6 alkoxy is optionally replaced by R 3a replace; R 3a is selected from D or halogen; m is selected from 0, 1, 2, 3, 4 or 5; n is selected from 0, 1 or 2.

2. The compound according to claim 1, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: Z1 is selected from N.

3. The compound according to claim 1 or 2, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: R a , R b , R c , R d , R e , R f Selected from H.

4. The compound according to claim 3, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: Selected from 5. The compound according to any one of claims 1 to 4, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: R 1 is selected from a 4-7 membered heterocyclyl, a C3-C6 cycloalkyl or a 5-6 membered heteroaryl, wherein the 4-7 membered heterocyclyl, the 3-6 membered cycloalkyl or the 5-6 membered heteroaryl is optionally replaced by R 1a Replace; or, R 1 is selected from pyridinyl, the pyridinyl being optionally substituted by R 1a replace.

6. The compound according to any one of claims 1 to 5, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: R 2 is selected from H, F or Cl.

7. The compound according to any one of claims 1 to 6, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: R 3 Selected from H or methyl.

8. The compound according to any one of claims 1 to 7, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: The compound of formula (I) is selected from the compound of formula (II) or its stereoisomer or a pharmaceutically acceptable salt thereof: Among them, R 1 , R 2 , R 3 , m and n are as defined in any one of 1 to 7.

9. The compound according to any one of claims 1 to 7, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: The compound of formula (I) is selected from the compound of formula (II) or its stereoisomer or a pharmaceutically acceptable salt thereof: Among them, R 1 , R 2 and m is as defined in any one of 1 to 7.

10. The compound according to any one of claims 1 to 9, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein: The compound of formula (I) is selected from the following compounds or their stereoisomers or pharmaceutically acceptable salts:

11. A pharmaceutical composition, wherein The pharmaceutical composition comprises the compound of formula (I) or its stereoisomer or pharmaceutically acceptable salt according to any one of claims 1 to 11, and pharmaceutically acceptable excipients.

12. Use of a compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof as described in any one of claims 1 to 10, or a pharmaceutical composition as described in claim 11, in the preparation of a medicament for treating or preventing a disease or condition associated with loss of TREM2 function in a mammal, preferably a human.

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