TREM2 agonists and uses thereof

Abstract

Disclosed herein are compounds of Formula (I) or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, as TREM2 agonists, methods of making the same, pharmaceutical compositions containing the same, and uses of the same in the prevention or treatment of diseases or disorders associated with loss of function of TREM2, including neurodegenerative diseases. Formula (I)

Description

[0001] This invention claims priority to two earlier applications: Patent Application No. 202510107777.6, filed with the China National Intellectual Property Administration on January 23, 2025, entitled "TREM2 Agonist and Its Use Thereof," and Patent Application No. 202510380219.7, filed with the China National Intellectual Property Administration on March 28, 2025, also entitled "TREM2 Agonist and Its Use Thereof." The entire contents of the aforementioned earlier applications are incorporated herein by reference. Technical Field

[0002] This invention belongs to the field of biomedicine and relates to TREM2 agonists, their preparation methods, pharmaceutical compositions containing the compounds, and their use in the prevention or treatment of diseases related to TREM2 loss of function. Background Technology

[0003] Microglia are innate immune cells found in the brain and are crucial for maintaining homeostasis in the central nervous system. As macrophages in the brain, these cells sense changes in the microenvironment and alter their phenotype through various receptors they express, mediating responses to invading pathogens, protein toxicity stress, and cell damage. Microglia can proliferate rapidly upon stimulation and are characterized by exhibiting bone marrow cell functions such as phagocytosis, cytokine / chemokine release, antigen presentation, and migration.

[0004] Triggering receptor 2 (TREM2), primarily expressed in microglia of the central nervous system (CNS), is involved in microglia proliferation, survival, migration, and phagocytosis. In mice lacking functional TREM2 expression or expressing a mutant receptor, a key observation is a diminished microglial response to injury, such as oligodendrocyte demyelination, stroke-induced brain damage, and in vivo protein toxicity inclusions. Pharmacological activation of TREM2 may serve as a potential target for the treatment or prevention of neurological disorders, neurodegenerative diseases, and other conditions. Summary of the Invention

[0005] This invention relates to compounds of formula (I) or their stereoisomers or pharmaceutically acceptable salts thereof.

[0006]

[0007] (I)

[0008] in,

[0009] X1 is selected from CR 5 Or N;

[0010] X2 is selected from CR 6 Or N;

[0011] R 1 Selected from phenyl or 5-6-membered heteroaryl, wherein the phenyl or 5-6-membered heteroaryl group is optionally R 1a replace;

[0012] R 1a Selected from halogens, OH, NH2, CN, C1-C6 alkyl, C3-C6 cycloalkyl, or 4-7 membered heterocyclic groups, wherein the OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, or 4-7 membered heterocyclic group is optionally replaced by R. 1b replace;

[0013] R 1b Selected from halogens, OH, NH2, CN, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy or C1-C6 haloalkoxy;

[0014] R 3 and R 4 Independently selected from H, C1-C6 alkyl, C3-C6 cycloalkyl, or 4-7 membered heterocyclic groups, wherein the C1-C6 alkyl, C3-C6 cycloalkyl, or 4-7 membered heterocyclic group is optionally replaced by R. 3a replace;

[0015] R 5 and R 6 Independently selected from H, halogen, OH, NH2, CN, 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. 5a replace;

[0016] R 3a and R 5a It is independently selected from D, halogen, OH, NH2, CN, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy or C1-C6 haloalkoxy;

[0017] R 2 Selected from or ;

[0018] Y1 is selected from CR 8 Or N;

[0019] Y2 is selected from C(R) 9 (R) 10 ) or C(O);

[0020] Y3 is selected from C(R) 11 (R) 12 );

[0021] Y4 is selected from C(R) 13 (R) 14 ), NR 15 O or S;

[0022] Y5 is selected from CR 16 Or N;

[0023] Y6 is selected from C(R) 17 (R) 18 );

[0024] Y7 is selected from C(R) 19 2. NR 20 O, S, or S=O;

[0025] Y9 is selected from key, C(R) 21 )2 or C=O;

[0026] Y 13 Selected from bond, O or C(R) 22 )2;

[0027] Y8, Y 10 Y 11 Y 12 Selected from C(R) 23 )2;

[0028] Y 14 Selected from CR 24 Or N;

[0029] m and n are independently selected from 0, 1, or 2;

[0030] R 7 Selected from C1-C6 alkyl, C1-C6 alkylamino, C1-C6 alkoxy, C1-C6 haloalkyl, -C(O)O-(C1-C6 alkyl), C3-C6 cycloalkyl, 4-7-membered heterocyclic, phenyl, -O-phenyl, 5-6-membered heteroaryl, or -O-(5-6-membered heteroaryl), wherein the C1-C6 alkyl, C1-C6 alkylamino, C1-C6 alkoxy, C1-C6 haloalkyl, -C(O)O-(C1-C6 alkyl), C3-C6 cycloalkyl, 4-7-membered heterocyclic, phenyl, -O-phenyl, 5-6-membered heteroaryl, or -O-(5-6-membered heteroaryl) is optionally R 7a replace;

[0031] R 7aSelected from O, halogen, CN, C1-C6 alkyl, C2-C6 alkenyl, C1-C6 alkoxy, C1-C6 alkylamino, C1-C6 haloalkyl, C3-C6 cycloalkyl, or 4-7 membered heterocyclic groups, wherein the C1-C6 alkyl, C2-C6 alkenyl, C1-C6 alkoxy, C1-C6 alkylamino, C1-C6 haloalkyl, C3-C6 cycloalkyl, or 4-7 membered heterocyclic group is optionally R 7b replace;

[0032] R 7b Selected from OH, halogen, C1-C6 alkyl, C1-C6 alkoxy or -C(O)O-(C1-C6 alkyl);

[0033] R 8 R 9 R 10 R 11 R 12 R 13 R 14 R 15 R 16 R 17 R 18 R 19 R 20 R 21 R 22 R 23 and R 24 Independently selected from H, D, halogen, OH, NH2, CN, C1-C6 alkyl, C3-C6 cycloalkyl, or 4-7 membered heterocyclic groups, wherein the OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, or 4-7 membered heterocyclic group is optionally replaced by R. 8a Replace; or, R 9 and R 10 R 11 and R 12 R 13 and R 14 Or R 17 and R 18 The atoms bonded to it together form a C3-C6 cycloalkyl or a 4-7 membered heterocyclic group, wherein the C3-C6 cycloalkyl or 4-7 membered heterocyclic group is optionally R 8b replace;

[0034] R 8a R 8b It is independently selected from D, halogen, OH, NH2, CN, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy or C1-C6 haloalkoxy.

[0035] In some implementations, X1 is selected from CH or N. In some implementations, X1 is selected from N.

[0036] In some implementations, X2 is selected from CH or N.

[0037] In some implementation schemes, R 1 Selected from phenyl, said phenyl optionally being R 1a replace.

[0038] In some implementation schemes, R 1a Selected from halogens or C1-C6 alkyl groups, wherein the C1-C6 alkyl group is optionally R 1b Replacement. In some implementations, R 1b Selected from halogens. In some embodiments, R 1a Selected from F, Cl or CF3.

[0039] In some implementation schemes, R 1 Selected from , or .

[0040] In some implementation schemes, R 3 Selected from H or C1-C6 alkyl groups, wherein the C1-C6 alkyl group is optionally converted to R 3a Replacement. In some implementations, R 3 Selected from H or C1-C3 alkyl groups, wherein the C1-C3 alkyl group is optionally converted to R 3a Replacement. In some implementations, R 3 Selected from H, CH3, CD3, CH2CH3, or CH(CH3)2. In some embodiments, R 3 Selected from H, CH3, CD3, or CH2CH3. In some embodiments, R 3 Selected from H, CH3 or CD3.

[0041] In some implementation schemes, R 4 Selected from H, C1-C6 alkyl or C3-C6 cycloalkyl, wherein the C1-C6 alkyl or C3-C6 cycloalkyl is optionally replaced by R. 3a Replacement. In some implementations, R 4 Selected from C1-C6 alkyl or cyclopropyl groups, wherein the C1-C6 alkyl or cyclopropyl group is optionally R 3a Replacement. In some implementations, R 4 Selected from CH3, CD3, CH2CH3, CH(CH3)2 or cyclopropyl.

[0042] In some implementation schemes, R 2 Selected from , or .

[0043] In some implementation schemes, R2 Selected from or .

[0044] In some implementations, Y1 is selected from CH or N.

[0045] In some implementations, Y2 is selected from CH2.

[0046] In some implementations, Y4 is selected from C(R) 13 (R) 14 Y4 is selected from O. In some implementations, Y4 is selected from O.

[0047] In some implementations, Y5 is selected from CH.

[0048] In some implementations, Y6 is selected from CH2.

[0049] In some implementation schemes, R 2 Selected from , , , or In some implementations, R 2 Selected from , , or In some implementations, R 2 Selected from , or .

[0050] In some implementation schemes, R 11 R 12 Independently selected from H, halogens, or C1-C6 alkyl groups, or R 11 and R 12 The atoms attached to it together form C3-C6 cycloalkyl groups.

[0051] In some implementation schemes, R 11 R 12 It is independently selected from H, halogens, or C1-C6 alkyl groups.

[0052] In some implementation schemes, R 11 R 12 It is independently selected from H or C1-C3 alkyl groups.

[0053] In some implementation schemes, R 11 R 12 Independently selected from H or CH3, or R 11 and R 12 The atoms bonded to it together form a cyclopropyl group.

[0054] In some implementation schemes, R 11 R 12 It is independently selected from H or CH3.

[0055] In some implementation schemes, R 13 R 14 It is independently selected from H or halogens.

[0056] In some implementation schemes, R 2 Selected from , , , , , , , or .

[0057] In some implementation schemes, R 2 Selected from , , , , , , or In some implementations, R 2 Selected from , , , , , or In some implementations, R 2 Selected from , , , or .

[0058] In some implementation schemes, R 7 Selected from phenyl or 5-6-membered heteroaryl, wherein the phenyl or 5-6-membered heteroaryl group is optionally R 7a Replacement. In some implementations, R 7 The group is selected from pyrazolyl, pyridyl, or pyrimidinyl, wherein the pyrazolyl, pyridyl, or pyrimidinyl group is optionally replaced by R. 7a Replacement. In some implementations, R 7 Selected from , or The , or Optionally R 7a replace.

[0059] In some implementation schemes, R 7a Selected from C1-C6 alkyl, C3-C6 cycloalkyl, or 4-7 membered heterocyclic groups, wherein the C1-C6 alkyl, C3-C6 cycloalkyl, or 4-7 membered heterocyclic group is optionally R 7b Replacement. In some implementations, R 7a Selected from C1-C6 alkyl or C3-C6 cycloalkyl. In some embodiments, R 7a Selected from C1-C3 alkyl or cyclopropyl groups. In some embodiments, R 7a Selected from methyl or cyclopropyl.

[0060] In some implementation schemes, R 7b Selected from halogens or C1-C6 alkoxy groups. In some embodiments, R 7b Selected from F or OCH3.

[0061] In some implementation schemes, R 7 Selected from , , , , , , , , , , or .

[0062] In some implementation schemes, R 7 Selected from , , or .

[0063] In some embodiments, the compound of formula (I) of the present invention, or its stereoisomer or a pharmaceutically acceptable salt thereof, is selected from the compound of formula (II) or its stereoisomer or a pharmaceutically acceptable salt thereof.

[0064]

[0065] (II)

[0066] Among them, R 1 R 2 R 3 and R 4 As defined in equation (I).

[0067] In some embodiments, the compound of formula (I) of the present invention, or its stereoisomer or a pharmaceutically acceptable salt thereof, is selected from the compound of formula (III) or its stereoisomer or a pharmaceutically acceptable salt thereof.

[0068]

[0069] (III)

[0070] Among them, R 1 R 2 R 3 and R 4 As defined in equation (I).

[0071] In some embodiments, the compound of formula (I) of the present invention, or its stereoisomer or a pharmaceutically acceptable salt thereof, is selected from the following compounds, their stereoisomers or pharmaceutically acceptable salts thereof:

[0072]

[0073] or .

[0074] On the other hand, the present invention provides a pharmaceutical composition comprising a compound of formula (I) of the present invention or a stereoisomer thereof or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

[0075] On the other hand, the present invention provides a method for treating diseases related to TREM2 loss of function in mammals, comprising administering to a mammal, preferably a human, a therapeutically effective amount of a compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0076] On the other hand, the present invention provides the use of a compound of formula (I) or its stereoisomer or pharmaceutically acceptable salt or pharmaceutical composition thereof in the preparation of a medicament for the prevention or treatment of diseases related to loss of TREM2 function.

[0077] On the other hand, the present invention provides the use of a compound of formula (I) or its stereoisomer or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, in the prevention or treatment of diseases related to loss of TREM2 function.

[0078] On the other hand, the present invention provides a compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof, for the prevention or treatment of diseases related to TREM2 loss of function.

[0079] In some implementations, the diseases associated with TREM2 function loss are selected from neurodegenerative diseases, preferably Alzheimer's disease.

[0080] Unless otherwise stated, the terminology used in this invention has the following meanings: the definitions of groups and terms described herein, including definitions as examples, exemplary definitions, preferred definitions, definitions listed in tables, definitions of specific compounds in the examples, etc., can be arbitrarily combined and combined with each other. A particular term should not be considered uncertain or unclear unless specifically defined, but should be understood according to its ordinary meaning in the art. When trade names appear herein, they are intended to refer to the corresponding product or its active ingredient.

[0081] In this article, " "" indicates a connection point.

[0082] The diagrammatic representation of racemic or enantiomerically pure compounds in this article is derived from Maehr, J. Chem. Ed. 1985, 62:114-120. Unless otherwise specified, wedge bonds and virtual wedge bonds are used. and ) represents the absolute configuration of a solid center.

[0083] The term "tautomer" refers to a functional group isomer resulting from the rapid movement of an atom between two positions within a molecule. The compounds of this invention can exhibit tautomerism. Tautomers can exist in two or more interconvertible forms. Tautomers generally exist in equilibrium; attempts to isolate a single tautomer typically produce a mixture whose physicochemical properties are consistent with those of the mixture of compounds. The equilibrium position depends on the intramolecular chemical characteristics. For example, in many aliphatic aldehydes and ketones such as acetaldehyde, the ketone form is dominant; while in phenols, the enol form is dominant. This invention encompasses all tautomeric forms of the compounds.

[0084] The term "stereoisomer" refers to isomers that are produced by different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers and diastereomers.

[0085] The compounds of this invention may have asymmetric atoms such as carbon, sulfur, nitrogen, and phosphorus atoms, or asymmetric double bonds, and therefore may exist in specific geometric or stereoisomer forms. Specific geometric or stereoisomer forms may be cis and trans isomers, E- and Z-type geometric isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)- isomers, (L)- isomers, and racemic mixtures thereof or other mixtures, such as mixtures enriched with enantiomers or diastereomers. All such isomers and mixtures thereof are within the scope of the definition of the compounds of this invention. Alkyl groups and other substituents may contain additional asymmetric carbon atoms, asymmetric sulfur atoms, asymmetric nitrogen atoms, or asymmetric phosphorus atoms. All such isomers involved in all substituents and mixtures thereof are also included within the scope of the definition of the compounds of this invention. The compounds containing asymmetric atoms of the present invention can be isolated in optically active pure form or in racemic form. The optically active pure form can be separated from racemic mixtures or synthesized using chiral raw materials or chiral reagents.

[0086] The term "substitution" refers to the substitution of one or more hydrogen atoms on a specific atom by a substituent, provided that the valence state of the specific atom is normal and the resulting compound is stable. When the substituent is oxo (i.e., =O), it means that two hydrogen atoms are substituted; oxo substitution does not occur on aromatic groups.

[0087] The terms “optional” or “optionally” mean that the event or condition subsequently described may or may not occur, including both the occurrence and non-occurrence of said event or condition. For example, “optionally” substituted with a halogen means that the ethyl group can be unsubstituted (CH2CH3), monosubstituted (CH2CH2F, CH2CH2Cl, etc.), polysubstituted (CHFCH2F, CH2CHF2, CHFCH2Cl, CH2CHCl2, etc.), or fully substituted (CF2CF3, CF2CCl3, CCl2CCl3, etc.). Those skilled in the art will understand that for any group containing one or more substituents, no substitution or substitution pattern that is spatially impossible and / or cannot be synthesized is introduced.

[0088] When any variable (e.g., R) a R b When a group appears more than once in the composition or structure of a compound, its definition is independent in each case. For example, if a group is surrounded by two R... b Replaced, then each R b Each has its own independent options.

[0089] When the number of a linking group is 0, such as -(CH2)0-, it indicates that the linking group is a bond.

[0090] When one of the variables is selected as a bond or does not exist, it means that the two groups it is connected to are directly connected. For example, when Y in AYZ represents a bond, it means that the structure is actually AZ.

[0091] C in this article m -C n It refers to having an integer number of carbon atoms in the range mn. For example, "C1-C 10 "" means that the group can have 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms.

[0092] The term "alkyl" refers to a compound with the general formula C10. n H 2n+1 The alkyl group can be straight-chain or branched. The term "C1-C6 alkyl" is understood to refer to a straight-chain or branched saturated monovalent hydrocarbon group having 1, 2, 3, 4, 5, or 6 carbon atoms, including but not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, neopentyl, hexyl, 2-methylpentyl, etc. The term "C1-C3 alkyl" is understood to refer to a straight-chain or branched saturated monovalent hydrocarbon group having 1 to 3 carbon atoms. The "C1-C6 alkyl" may further include "C1-C3 alkyl".

[0093] The term "alkoxy" refers to a monovalent group formed by the loss of a hydrogen atom from a hydroxyl group in straight-chain or branched alcohols. The term "C1-C6 alkoxy" may further include "C1-C3 alkoxy".

[0094] The term "alkylamino" refers to a monovalent group formed by the loss of a hydrogen atom from the amino group of a straight-chain or branched amine. The term "C1-C6 alkylamino" may further include "C1-C3 alkylamino".

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

[0096] The term "halogenated alkyl" includes monohalogenated or polyhalogenated alkyl. The term "C1-C6 haloalkyl" may further include "C1-C3 haloalkyl".

[0097] The term "haloalkoxy" includes monohaloalkoxy or polyhaloalkoxy. The term "C1-C6 haloalkoxy" may further include "C1-C3 haloalkoxy".

[0098] The term "haloalkylamino" includes monohalogenated or polyhalogenated alkylamino. The term "C1-C6 haloalkylamino" may further include "C1-C3 haloalkylamino".

[0099] The term "cycloalkyl" refers to a fully saturated carbon ring existing in the form of a monocyclic, fused, bridged, or spirocyclic ring. Unless otherwise indicated, the carbon ring is typically a 3- to 12-membered ring. The term "C3-C" is also used. 12 "Cycloalkyl" should be understood as referring to a saturated monovalent monocyclic, fused, spirocyclic, or bridged ring having 3 to 12 carbon atoms. Specific examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, norbornyl (bicyclo[2.2.1]heptyl), bicyclo[2.2.2]octyl, adamantyl, spiro[4.5]decyl, etc. The term "C3-C" 12 "Cycloalkyl" can include "C3-C6 cycloalkyl". The term "C3-C6 cycloalkyl" can be understood as indicating a saturated 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.

[0100] The term "heterocyclic group" refers to a fully saturated or partially saturated (not aromatic as a whole) monovalent monocyclic, fused, spirocyclic, or bridged ring group containing 1 to 5 heteroatoms or heteroatom groups (i.e., groups containing heteroatoms). The "heteroatoms or heteroatom groups" include, but are not limited to, nitrogen (N), oxygen (O), sulfur (S), phosphorus (P), boron (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 with 4, 5, 6, 7, 8, 9, 10, 11 or 12 ring atoms, and its ring atoms contain 1 to 5 heteroatoms or heterogroups independently selected from those described above. "4-12-membered heterocyclic groups" include "4-7-membered heterocyclic groups", wherein specific examples of 4-membered heterocyclic groups include, but are not limited to, azacyclic butyl or oxacyclic butyl; specific examples of 5-membered heterocyclic groups include, but are not limited to, tetrahydrofuranyl, dioxacyclopentenyl, pyrrolyl, imidazoyl, pyrazolyl, pyrrolinyl, 4,5-dihydrooxazolyl or 2,5-dihydro-1H-pyrrolyl; specific examples of 6-membered heterocyclic groups include, but are not limited to, tetrahydropyranyl, piperidinyl, morpholinyl, dithiaalkyl, thiomorpholinyl, piperazinyl, trithiaalkyl, tetrahydropyridinyl or 4H-[1,3,4]thiadiazinyl; and specific examples of 7-membered heterocyclic groups include, but are not limited to, diazacyclic heptyl. The heterocyclic group can also be a bicyclic group, wherein specific examples of 5,5-membered bicyclic groups include, but are not limited to, hexahydrocyclopentano[c]pyrrolo-2(1H)-yl; specific examples of 5,6-membered bicyclic groups include, but are not limited to, hexahydropyrrolo[1,2-a]pyrazin-2(1H)-yl, 5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazinyl, or 5,6,7,8-tetrahydroimidazo[1,5-a]pyrazinyl. Optionally, the heterocyclic group can be a benzofused cyclic group of the above-mentioned 4-7-membered heterocyclic groups, specific examples of which include, but are not limited to, dihydroisoquinolinyl, etc. The term "4-12 membered heterocyclic group" can include the ranges of "5-12 membered heterocyclic group", "4-7 membered heterocyclic group", "5-6 membered heterocyclic group", "6-8 membered heterocyclic group", "4-12 membered heterocyclic alkyl group", "5-12 membered heterocyclic alkyl group", "4-7 membered heterocyclic alkyl group", "5-6 membered heterocyclic alkyl group", and "6-8 membered heterocyclic alkyl group". "4-7 membered heterocyclic group" can further include the ranges of "4-6 membered heterocyclic group", "5-6 membered heterocyclic group", "4-7 membered heterocyclic alkyl group", "4-6 membered heterocyclic alkyl group", and "5-6 membered heterocyclic alkyl group". Although some bicyclic heterocyclic groups in this invention partially contain a benzene ring or a heteroaromatic ring, the heterocyclic group as a whole is still non-aromatic.

[0101] The term "heteroaryl" refers to an aromatic monocyclic or fused polycyclic system containing at least one ring atom selected from N, O, or S, with the remaining ring atoms being C. The term "5-10-membered heteroaryl" should be understood to include monovalent monocyclic or bicyclic aromatic ring systems having 5, 6, 7, 8, 9, or 10 ring atoms, particularly 5, 6, 9, or 10 ring atoms, and containing 1-5, preferably 1-3, heteroatoms independently selected from N, O, and S. Specifically, the heteroaryl group is selected from thienyl, furanyl, pyrroleyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl or thiadiazolyl and their benzo[derivatives], such as benzofuranyl, benzothienyl, benzothiazolyl, benzooxazolyl, benzoisooxazolyl, benzoimidazolyl, benzotriazolyl, indazole, indolyl or isindolyl; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl or triazinyl and their benzo[derivatives], such as quinolinyl, quinazolinyl or isoquinolinyl; or acrylinyl, inazinyl, purinyl and their benzo[derivatives]; or cyclolinyl, phthalazinyl, quinazolinyl, quinoxolinyl, naphthidyl, pteridinyl, carbazolyl, acrylinyl, phenazinyl, phenothiazinyl or phenothiazinyl. The term "5-6 heteroaryl" refers to an aromatic ring system having 5 or 6 ring atoms, and containing 1 to 3, preferably 1 to 2, heteroatoms independently selected from N, O and S.

[0102] The term "therapeutic effective amount" means (i) the amount of the compound of the present invention used to treat or prevent a particular disease, condition, or disorder; (ii) to reduce, improve, or eliminate one or more symptoms of a particular disease, condition, or disorder; or (iii) to prevent or delay 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 "therapeutic effective amount" varies depending on the compound, the disease state and its severity, the route of administration, and the age of the mammal to be treated, but may routinely be determined by those skilled in the art based on their own knowledge and the present disclosure.

[0103] The term "pharmaceutical acceptable" refers to compounds, materials, compositions, and / or dosage forms that, within the bounds of reliable medical judgment, are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0104] The term "pharmaceutically acceptable salt" refers to a pharmaceutically acceptable salt of an acid or base, including salts formed by a compound with an inorganic or organic acid, and salts formed by a compound with an inorganic or organic base.

[0105] The term "pharmaceutical composition" refers to a mixture of one or more compounds of the present invention or their salts with pharmaceutically acceptable excipients. The purpose of a pharmaceutical composition is to facilitate the administration of the compounds of the present invention to an organism.

[0106] The term "pharmaceuticalally acceptable excipient" refers to excipients that do not cause significant irritation to the organism and do not impair the biological activity and properties of the active compound. Suitable excipients are well known to those skilled in the art, such as carbohydrates, waxes, water-soluble and / or water-swellable polymers, hydrophilic or hydrophobic materials, gelatin, oils, solvents, water, etc.

[0107] The word “comprise” or “comprise” and its English variants such as comprises or comprising should be understood in an open, non-exclusive sense, meaning “including but not limited to”.

[0108] This invention also includes compounds of the invention that are identical to those described herein, but in which one or more atoms are labeled with isotopes whose atomic weights or mass numbers differ from those commonly found in nature. Examples of isotopes that can be incorporated into the compounds of the 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.

[0109] Certain isotope-labeled compounds of the present invention (e.g., using...) 3 H and 14 C-labeling can be used in the analysis of compound and / or substrate tissue distribution. Tritiumization (i.e., 3 H) and carbon-14 (i.e. 14 C) Isotopes are particularly preferred due to their ease of preparation and detectability. Positron-emitting isotopes, such as... 15 O、 13 N、 11 C and 18F can be used in positron emission tomography (PET) studies to determine substrate occupancy. The isotopically labeled compounds of the present invention can typically be prepared by replacing the unlabeled reagent with an isotopically labeled reagent using a procedure similar to those disclosed in the schemes and / or examples below.

[0110] The pharmaceutical compositions of the present invention can be prepared by combining the compounds of the present invention with suitable pharmaceutically acceptable excipients, for example, in solid, semi-solid, liquid or gaseous formulations, such as tablets, pills, capsules, powders, granules, ointments, emulsions, suspensions, suppositories, injections, inhalers, gels, microspheres and aerosols.

[0111] Typical routes of administration of the compounds of the present invention or their pharmaceutically acceptable salts or pharmaceutical compositions thereof include, but are not limited to, oral, rectal, topical, inhalation, parenteral, sublingual, vaginal, intranasal, intraocular, intraperitoneal, intramuscular, subcutaneous, and intravenous administration.

[0112] The pharmaceutical compositions of the present invention can be manufactured using methods well known in the art, such as conventional mixing, dissolving, granulation, emulsification, freeze drying, etc.

[0113] 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, sugar-coated tablets, capsules, liquids, gels, pastes, suspensions, etc., for oral administration to patients.

[0114] Solid oral compositions can be prepared using conventional mixing, filling, or tableting methods. For example, they can be obtained by mixing the active compound with solid excipients, optionally milling the resulting mixture, adding other suitable excipients if necessary, and then processing the mixture into granules to obtain the core of a tablet or sugar-coated formulation. Suitable excipients include, but are not limited to, binders, diluents, disintegrants, lubricants, flow aids, or flavoring agents.

[0115] The pharmaceutical composition may also be suitable for parenteral administration, such as in suitable unit dosage forms of sterile solutions, suspensions or lyophilized products.

[0116] In all methods of administration of the compound of general formula I described herein, the daily dose 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 separate doses.

[0117] The compounds of the present invention can be prepared by a variety of synthetic methods known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthetic methods, and equivalent substitutions known to those skilled in the art. Preferred embodiments include, but are not limited to, the embodiments of the present invention.

[0118] The chemical reactions in the specific embodiments of this invention are carried out in a suitable solvent, which must be suitable for the chemical changes of this invention and the reagents and materials required therefor. To obtain the compounds of this invention, it is sometimes necessary for those skilled in the art to modify or select the synthesis steps or reaction flow based on existing embodiments.

[0119] This invention uses the following abbreviations:

[0120] DCM: dichloromethane; TfOH: trifluoromethanesulfonic acid; Pd(dppf)Cl2: [1,1'-bis(diphenylphosphine)ferrocene]palladium(II) dichloride; KOAc: potassium acetate; Dioxane: 1,4-dioxane; PtO2: platinum dioxide; THF: tetrahydrofuran; DMF: N,N-dimethylformamide; Pd(dppf)Cl2·DCM: [1,1'-bis(diphenylphosphine)ferrocene]palladium(II) dichloride dichloromethane complex; HATU: 2-(7-azabenzo[] Triazole)-N,N,N',N'-Tetramethylurea hexafluorophosphate; DIPEA: N,N-diisopropylethylamine; DMSO: Dimethyl sulfoxide; B2Pin2: Pinacol diboronate; EtOH: Ethanol; Pyridinium triribromide: Pyridinium tribromide; TEA: Triethylamine; TsCl: p-Toluenesulfonyl chloride; TES: Triethoxysilane; TMSOTf: Trimethylsilyl trifluoromethanesulfonate; DCE: Dichloroethane; DAST: Diethylaminosulfonium trifluoride; XPhos Pd G4: Methanesulfonic acid (2-dicyclohexylphosphino-2',4',6'-tri-isopropyl-1,1'-biphenyl)(2'-methylamino-1,1'-biphenyl-2-yl)palladium(II); DIC: N,N'-diisopropylcarbodiimide; DMAP: 4-dimethylaminopyridine; TMSCl: trimethylchlorosilane; DMA: N,N-dimethylacetamide. Detailed Implementation

[0121] The invention is described in detail below with reference to embodiments, but this does not imply any adverse limitation on the invention. The invention has been described in detail herein, and specific embodiments thereof have also been disclosed. It will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of the invention without departing from the spirit and scope thereof. All reagents used in this invention are commercially available and can be used without further purification.

[0122] Unless otherwise stated, proportions expressed for mixed solvents are volume-based. Unless otherwise stated, % refers to wt%.

[0123] Compounds are named manually or using ChemDraw® software; commercially available compounds are named using the supplier catalog name.

[0124] The structure of the compound was determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). NMR shifts are measured in units of 10⁻⁶. -6 (ppm). The solvents used for NMR determination were deuterated dimethyl sulfoxide, deuterated chloroform, deuterated methanol, etc., and the internal standard was tetramethylsilane (TMS).

[0125] Example 1: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholine)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (Compound 1)

[0126]

[0127] Step 1: Synthesis of ethyl 5-amino-2-chloro-6-(4-chloro-2-fluorophenyl)pyrimidine-4-carboxylate (1-3)

[0128] To a solution of 1,4-dioxane in water (50 mL: 5 mL) containing 1-1 (2 g, 8.47 mmol) and 1-2 (1.47 g, 8.47 mmol), Pd(dppf)Cl2 (1.23 g, 1.68 mmol) and K3PO4 (5.4 g, 25.4 mmol) were added, and the mixture was stirred at 55 °C for 3 hours under N2 protection. The resulting mixture was diluted with water and EtOAc and extracted three times with EtOAc. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. Purification was performed on a silica gel column, eluting with 800%–300% PE / EA solution to give 1-3 (1.6 g).

[0129] 1 H NMR (300 MHz, DMSO-d6) d 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).

[0130] Step 2: Synthesis of 5-amino-2-chloro-6-(4-chloro-2-fluorophenyl)pyrimidine-4-carboxylic acid (1-4)

[0131] To a solution of 1-3 (1.8 g, 5.47 mmol) in 30 mL of THF, a solution of LiOH (1.15 g, 27.4 mmol) in 20 mL of H₂O was added. The mixture was stirred at room temperature for 2 hours. The reaction was quenched with water and 30 mL of 2 M HCl aqueous solution, and the mixture was extracted three times with DCM. The combined organic layers were dried over anhydrous Na₂SO₄ and concentrated under vacuum to give the crude product 1-4 (1.6 g).

[0132] Step 3: Synthesis of 5-amino-2-chloro-6-(4-chloro-2-fluorophenyl)-N-methylpyrimidine-4-carboxamide (1-5)

[0133] 1-4 (1.27 g, 4.2 mmol), methylamine hydrochloride (428 mg, 6.3 mmol), and HATU (2.4 g, 6.3 mmol) were stirred in DMF (30 mL) at 20°C under nitrogen, and DIPEA (1.63 g, 12.7 mmol) was added. The reaction mixture was stirred at 50°C for 2 hours. The reaction mixture was diluted with H2O (100 mL) and extracted with EtOAc. The combined organic matter was dried over anhydrous MgSO4, filtered, and concentrated under vacuum to give the residue. The residue was purified by elution of 800%–300% PE / EA solution onto a silica gel column to give 1-5 (1.24 g).

[0134] 1 H NMR (300 MHz, DMSO-d6) d 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).

[0135] Step 4: Synthesis of 6-chloro-8-(4-chloro-2-fluorophenyl)-3-methylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (1-6)

[0136] Dissolve 1-5 (250 mg, 0.79 mmol) in 10 mL of anhydrous tetrahydrofuran solution. Add NaH (80 mg, 3.33 mmol) at 0 °C and react at room temperature for 5 min. Then add triphosgene (250 mg, 0.84 mmol) and react at room temperature for 10 h. After the reaction is complete, quench the crude reaction with saturated sodium bicarbonate solution, extract three times with ethyl acetate, dry the organic layer with MgSO4, filter and concentrate. Purify on a silica gel column, elute with 800%–200% PE / EA solution, and separate to obtain 1-6 (130 mg).

[0137] 1H NMR (300 MHz, DMSO-d6) δ 11.53 (s, 1H), 7.71 (dt, J = 10.1, 2.8Hz, 1H), 7.61 (t, J = 8.0 Hz, 1H), 7.50 (dd, J = 8.3, 1.9 Hz, 1H), 3.27 (s,3H).

[0138] Step 5: Synthesis of 6-chloro-8-(4-chloro-2-fluorophenyl)-1,3-dimethylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (1-7)

[0139] Dissolve 1-6 (100 mg, 0.29 mmol) in 10 mL of anhydrous DMF solution. Then add NaH (11 mg, 0.44 mmol) at 0 °C and react at room temperature for 5 min. Then add CH3I (138 mg, 59.5 μL, 0.96 mmol) and react at room temperature for 3 h. After the reaction is complete, quench the reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, elute with 800%–400% PE / EA solution to obtain 1-7 (90 mg).

[0140] 1 H NMR (300 MHz, Chloroform-d) δ 7.73 (t, J = 8.0 Hz, 1H), 7.40 (ddd,J = 8.3, 2.0, 0.7 Hz, 1H), 7.29 – 7.25 (m, 1H), 3.54 (s, 3H), 3.13 (d, J =0.7 Hz, 3H).

[0141] Step 6: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholine)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (Compound 1)

[0142] Dissolve compounds 1-7 (100 mg, 0.2 mmol) in 10 mL of DMSO solution. Add DIPEA (0.5 mL) and compounds 1-8 (40 mg, 0.24 mmol), and react at 80 °C for 3 hours. Quench the reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, elute with 800%–400% PE / EA solution, and separate to obtain compound 1 (24 mg).

[0143] 1H NMR (400 MHz, DMSO-d6) δ 7.81 – 7.71 (m, 2H), 7.64 (dd, J = 10.0, 2.1 Hz, 1H), 7.49 (dd, J = 8.3, 2.0 Hz, 1H), 7.44 (s, 1H), 4.54 – 4.44 (m,2H), 4.39 (d, J = 13.2 Hz, 1H), 4.05 – 3.94 (m, 1H), 3.81 (s, 3H), 3.63 (td,J = 11.6, 2.8 Hz, 1H), 3.29 (s, 3H), 3.20 – 3.07 (m, 2H), 2.86 (s, 3H).

[0144] MS m / z (ES-API+): 486.10 [M+H] +

[0145] Example 2: Synthesis of 8-(4-chloro-2-fluorophenyl)-6-(2-(1-cyclopropyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-4-yl)-1,3-dimethylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 2)

[0146]

[0147] Step 1: Synthesis of 6-(1-cyclopropyl-1H-pyrazol-4-yl)-3,6-dihydro-2H-pyran-4-yltrifluoromethanesulfonate (2-2)

[0148] Under nitrogen protection, (2-hydroxyethyl)acetylene (1.9 g, 27.2 mmol) and DCM (40 mL) were added to 2-1 (2.47 g, 18.18 mmol). Then, TfOH (3.3 g, 21.8 mmol) was slowly added dropwise to the flask at 0 °C. After reacting for 5 minutes, the reaction was heated to room temperature. After 5 hours, TfOH (3.3 g, 21.8 mmol) was added dropwise again, and the reaction continued for 18 hours. After the reaction was complete, the crude reaction was quenched with saturated sodium bicarbonate aqueous solution, extracted three times with DCM, and the organic layer was dried over anhydrous MgSO4, filtered, and concentrated. Purification was performed on a silica gel column, eluted with 800%–100% PE / EA solution, to obtain 2-2 (1 g).

[0149] Step 2: Synthesis of 1-cyclopropyl-4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-5,6-dihydro-2H-pyran-2-yl)-1H-pyrazole (2-3)

[0150] 2-2 (245 mg, 0.727 mmol), Pd(dppf)Cl2·DCM (59.4 mg, 0.073 mmol), B2Pin2 (277 mg, 1.09 mmol), and potassium acetate (285 mg, 2.91 mmol) were added to 1,4-dioxane (2.9 mL) under nitrogen protection. The reaction mixture was heated to 90 °C and reacted for 2 hours. After the reaction was completed, the mixture was cooled to room temperature. The reaction was quenched with water, and the mixture was extracted three times with ethyl acetate. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. The mixture was purified on a silica gel column and eluted with 800%–100% PE / EA solution to give 2-3 (56 mg).

[0151] 1 H NMR (300 MHz, Chloroform-d) δ 7.49 (d, J = 0.8 Hz, 1H), 7.45 (s,1H), 6.58 (q, J = 2.1 Hz, 1H), 5.18 (q, J = 2.8 Hz, 1H), 3.91 (dt, J = 11.3,4.8 Hz, 1H), 3.73 (ddd, J = 11.5, 7.4, 4.5 Hz, 1H), 3.58 (tt, J = 7.3, 3.8Hz, 1H), 2.37 – 2.21 (m, 2H), 1.29 (s, 12H), 1.14 – 1.10 (m, 2H), 1.05 – 0.99(m, 2H).

[0152] Step 3: Synthesis of 8-(4-chloro-2-fluorophenyl)-6-(6-(1-cyclopropyl-1H-pyrazol-4-yl)-3,6-dihydro-2H-pyran-4-yl)-1,3-dimethylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (2-4)

[0153] 1-7 (70 mg, 0.2 mmol), 2-3 (45 mg, 0.24 mmol), Pd(dppf)Cl2·DCM (14 mg, 0.016 mmol), and potassium carbonate (83 mg, 0.6 mmol) were added to 1,4-dioxane and water (5 mL: 1 mL) under nitrogen protection. The reaction was carried out at 80 °C for 5 hours. After the reaction was complete, the reaction was quenched with water, and the mixture was extracted three times with ethyl acetate. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. The mixture was purified on a silica gel column and eluted with 800%–100% PE / EA solution to obtain 2-4 (30 mg).

[0154] Step 4: Synthesis of 8-(4-chloro-2-fluorophenyl)-6-(2-(1-cyclopropyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-4-yl)-1,3-dimethylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 2)

[0155] Ethanol (5 mL) was added to compound 2-4 (30 mg, 0.059 mmol) and PtO2 (13 mg, 0.055 mmol), and the reaction was carried out at room temperature for 3 hours under a hydrogen atmosphere. After the reaction was complete, the mixture was filtered, and the residue was washed with a small amount of ethanol. The filtrate was concentrated and then dry-coated onto a silica gel column for chromatography, eluting with 400%–10% PE / EA solution to give compound 2 (18 mg).

[0156] 1 H NMR (300 MHz, Chloroform-d) δ 7.70 (t, J = 8.0 Hz, 1H), 7.44 (d, J= 2.3 Hz, 2H), 7.38 (dd, J = 8.3, 1.9 Hz, 1H), 7.28 – 7.21 (m, 1H), 4.49 (dd,J = 11.4, 2.1 Hz, 1H), 4.21 (ddd, J = 11.6, 4.3, 2.1 Hz, 1H), 3.74 (td, J =11.5, 3.3 Hz, 1H), 3.54 (s, 3H), 3.53 – 3.40 (m, 2H), 3.12 (s, 3H), 2.36 –1.93 (m, 4H), 1.15 – 0.91 (m, 4H).

[0157] MS m / z (ES-API+): 511.20 [M+H] +

[0158] Example 3: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-((2R)-2-methyl-6-(1-methyl-1H-pyrazol-4-yl)morpholinyl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 3)

[0159]

[0160] Step 1: Synthesis of 2-bromo-1-(1-methyl-1H-pyrazol-4-yl)ethane-1-one (3-2)

[0161] 3-1 (100 mg, 0.81 mmol) and pyridinium tribromide (259 mg, 0.81 mmol) were dissolved in 5 mL of a mixed solvent (DCM:EtOH = 4:1) and reacted at room temperature for 12 hours. After the reaction was complete, the reaction mixture was diluted with an aqueous sodium sulfite solution, extracted three times with ethyl acetate, and the organic layer was dried over anhydrous MgSO4, filtered, and concentrated. Purification was performed on a silica gel column, eluting with 800%–600% PE / EA solution to give 3-2 (127 mg).

[0162] Step 2: Synthesis of (R)-N-(2-hydroxypropyl)-4-methylbenzenesulfonamide (3-4)

[0163] Anhydrous dichloromethane solution (40 mL) and triethylamine (6 g, 60 mmol) were added to 3-3 (1.5 g, 20 mmol), followed by the addition of p-toluenesulfonyl chloride (3.8 g, 20 mmol) under ice bath conditions. After the addition was complete, the mixture was heated to room temperature and reacted for 4 hours at room temperature. After the reaction was complete, the mixture was extracted three times with DCM, and the organic layer was dried over anhydrous MgSO4, filtered, and concentrated. The solution was purified on a silica gel column and eluted with 800%–300% PE / EA solution to obtain 3-4 (424 mg).

[0164] Step 3: Synthesis of (R)-N-(2-hydroxypropyl)-4-methyl-N-(2-(1-methyl-1H-pyrazol-4-yl)-2-oxoethyl)benzenesulfonamide (3-5)

[0165] To a 4 mL solution of acetone containing 3-4 (84.6 mg, 0.369 mmol), 3-2 (50 mg, 0.246 mmol), potassium carbonate (102 mg, 0.728 mmol), and potassium iodide (41 mg, 0.246 mmol) were added, and the mixture was reacted at room temperature for 8 hours. After the reaction was complete, the reaction was quenched with water, and the mixture was extracted three times with ethyl acetate. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. The solution was purified on a silica gel column and eluted with 800%–10% PE / EA solution to obtain 3-5 (81 mg).

[0166] Step 4: Synthesis of (2R)-2-methyl-6-(1-methyl-1H-pyrazol-4-yl)-4-toluenesulfonylmorpholine (3-6)

[0167] Under nitrogen atmosphere and at 0°C, TES (115 mg, 1 mmol) and TMSOTf (220 mg, 1 mmol) were added to a 7 mL solution of DCM containing 3-5 (70 mg, 0.19 mmol). After the addition was complete, the mixture was heated to room temperature and reacted for 12 hours. After the reaction was complete, the reaction was quenched with water, and the mixture was extracted three times with dichloromethane. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. The solution was purified on a silica gel column and eluted with 800%–10% PE / EA solution to obtain 3-6 (60 mg).

[0168] Step 5: Synthesis of (2R)-2-methyl-6-(1-methyl-1H-pyrazol-4-yl)morpholine (3-7)

[0169] Magnesium powder and magnesium shavings (200 mg, 8.3 mmol) were activated with 1 M HCl solution, and the residue was obtained by filtration. The activated magnesium powder and magnesium shavings were added to 8 mL of a methanol solution (85 mg, 0.25 mmol) containing magnesium 3-6, and the mixture was reacted at 80 °C for 12 hours. After the reaction was complete, the reaction solution was cooled to room temperature, filtered with diatomaceous earth, and the residue was washed with a small amount of methanol. The filtrates were combined and concentrated to obtain crude product 3-7 (20 mg), which was used directly in the next step.

[0170] Step 6: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-((2R)-2-methyl-6-(1-methyl-1H-pyrazol-4-yl)morpholinyl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 3)

[0171] Dissolve compound 1-7 (117 mg, 0.33 mmol) in DMSO (10 mL). Add DIPEA (1 mL) and 3-7 (90 mg, 0.49 mmol), and react at 80 °C for 3 hours. Quench the reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, elute with 800%–100% PE / EA solution, and separate to obtain compound 3 (45 mg).

[0172] 1H NMR (300 MHz, Chloroform-d) δ 7.56 (d, J = 11.9 Hz, 2H), 7.45 (s,1H), 7.33 (d, J = 7.3 Hz, 1H), 7.25 – 7.16 (m, 1H), 4.77 (d, J = 11.7 Hz,1H), 1.31 (d, J = 6.0Hz, 3H).

[0173] MS m / z (ES-API+): 500.20 [M+H] +

[0174] Example 4: Synthesis of 8-(4-chloro-2-fluorophenyl)-3-methyl-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholine)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 4)

[0175]

[0176] Dissolve compound 1-6 (100 mg, 0.29 mmol) in DMSO (10 mL). Add DIPEA (1 mL) and 1-8 (60 mg, 0.46 mmol), and react at 80 °C for 3 hours. Quench the crude reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, elute with 800%–5% PE / EA solution, and separate to obtain compound 4 (24 mg).

[0177] 1H NMR (300 MHz, DMSO-d6) δ 10.91 (s, 1H), 7.73 (s, 1H), 7.65 – 7.60 (m, 1H), 7.60 – 7.54 (m, 1H), 7.46 (d, J = 2.0 Hz, 1H), 7.43 (d, J = 0.8 Hz,1H), 4.49 (d, J = 2.8 Hz, 1H), 4.45 (d, J = 3.5 Hz, 1H), 4.36 (d, J = 13.1Hz, 1H), 3.97 (d, J = 10.1 Hz, 1H), 3.80 (s, 3H), 3.67 – 3.58 (m, 1H), 3.23(d, J = 6.0 Hz, 3H), 3.12 (d, J = 15.7 Hz, 1H), 3.06 – 3.00 (m, 1H).

[0178] MS m / z (ES-API+): 472.10 [M+H] +

[0179] Example 5: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-(2-(2-methylpyrimidin-4-yl)-5-oxa-2,8-diazaspiro[3.5]nonane-8-yl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 5)

[0180]

[0181] Step 1: Synthesis of tert-butyl 8-(4-(4-chloro-2-fluorophenyl)-5,7-dimethyl-6,8-dioxo-5,6,7,8-tetrahydropyrimidino[5,4-d]pyrimidin-2-yl)-5-oxa-2,8-diazaspiro[3,5]nonane-2-carboxylic acid (5-2)

[0182] Dissolve 1-7 (80 mg, 0.24 mmol) in DMSO (10 mL). Add DIPEA (0.4 mL) and 5-1 (77 mg, 0.36 mmol), and react at 100 °C for 2 hours. Quench the reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, elute with 800%–100% PE / EA solution, and separate to obtain 5-2 (97 mg).

[0183] Step 2: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-(5-oxa-2,8-diazaspiro[3.5]non-8-yl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione hydrochloride (5-3)

[0184] Add 0.5 mL of trifluoroacetic acid to 8 mL of DCM solution 5-2 (97 mg, 0.18 mmol) and react at room temperature for 2 hours. After the reaction is complete, concentrate directly to obtain crude product 5-3 (93.9 mg).

[0185] Step 3: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-(2-(2-methylpyrimidin-4-yl)-5-oxa-2,8-diazaspiro[3.5]nonane-8-yl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 5)

[0186] Dissolve compound 5-3 (93.9 mg, 0.21 mmol) in DMSO (10 mL). Add DIPEA (0.2 mL) and 5-4 (18 mg, 0.21 mmol), and react at 100 °C for 2 hours. Quench the reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 800%–5% PE / EA solution to obtain compound 5 (45 mg).

[0187] 1 H NMR (300 MHz, Chloroform-d) δ 8.11 (d, J = 6.0 Hz, 1H), 7.56 (t, J= 8.0 Hz, 1H), 7.32 (dd, J = 8.2, 2.0 Hz, 1H), 7.25 – 7.18 (m, 1H), 6.05 (d,J = 6.0 Hz, 1H), 4.00 (s, 2H), 3.82 (d, J = 4.5 Hz, 2H), 3.77 – 3.73 (m, 2H), 3.65 – 3.61 (m, 2H), 3.51 (s, 3H), 3.05 (s, 3H), 2.61 (s, 2H), 2.52 (s, 3H).

[0188] MS m / z (ES-API+): 539.20 [M+H] +

[0189] Example 6: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-(2-(1-methyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-4-yl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 6)

[0190]

[0191] Step 1: Synthesis of 6-(1-methyl-1H-pyrazol-4-yl)-3,6-dihydro-2H-pyran-4-yltrifluoromethanesulfonate (6-2)

[0192] Under nitrogen protection, (2-hydroxyethyl)acetylene (1.9 g, 27.2 mL) and DCM (40 mL) were added to 6-1 (2 g, 18.18 mmol). Then, TfOH (3.3 g, 21.8 mmol) was slowly added dropwise to the flask at 0 °C. After reacting for 5 minutes, the reaction was heated to room temperature. After 5 hours, TfOH (3.3 g, 21.8 mmol) was added dropwise again, and the reaction continued for 18 hours. After the reaction was complete, the reaction was quenched with saturated sodium bicarbonate solution, extracted three times with DCM, and the organic layer was dried over anhydrous MgSO4, filtered, and concentrated. Purification was performed on a silica gel column, eluted with 800%–100% PE / EA solution, to obtain 6-2 (2.3 g).

[0193] 1 H NMR (300 MHz, Chloroform-d) δ ppm 7.49 (s, 1 H), 7.37 (s, 1 H), 5.96 (dt, 7=2.6, 1.4 Hz, 1 H), 5.34 (q, J=2.6 Hz, 1 H), 3.98 - 4.04 (m, 1 H),3.92 (s, 3 H), 3.85 (ddd, 7=11.5, 6.4, 5.2 Hz, 1 H), 2.45 - 2.60 (m, 2 H).

[0194] Step 2: Synthesis of 1-methyl-4-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-5,6-dihydro-2H-pyran-2-yl)-1H-pyrazole (6-3)

[0195] 6-2 (227 mg, 0.727 mmol), Pd(dppf)Cl2·DCM (59.4 mg, 0.073 mmol), B2Pin2 (277 mg, 1.09 mmol), and potassium acetate (285 mg, 2.91 mmol) were added to 1,4-dioxane (2.9 mL) under nitrogen protection. The reaction mixture was heated to 90 °C and reacted for 2 hours. After the reaction was completed, the mixture was cooled to room temperature. The reaction was quenched with water, and the mixture was extracted three times with ethyl acetate. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. The mixture was purified on a silica gel column and eluted with 800%–100% PE / EA solution to give 6-3 (87 mg).

[0196] 1 H NMR (300 MHz, Chloroform-d)δ ppm 7.48 (s, 1H), 7.36 (s, 1H), 6.61 (q, J=1.9 Hz, 1H), 5.20 (q, J=2.6 Hz, 1H), 3.89-3.93 (m, 1H), 3.89 (s, 3 H), 3.71-3.78 (m, 1H), 2.28-2.39 (m, 1H), 2.17-2.27 (m, 1 H), 1.30 (s, 12H).

[0197] Step 3: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-(6-(1-methyl-1H-pyrazol-4-yl)-3,6-dihydro-2H-pyran-4-yl)pyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (6-4)

[0198] To a solution of 1,4-dioxane (100 mg, 0.29 mmol) and 6-3 (56 mg, 0.35 mmol) in water (5 mL: 1 mL), Pd(dppf)₂Cl₂·DCM (19 mg, 0.023 mmol) and KOAc (85 mg, 0.87 mmol) were added, and the mixture was stirred at 80 °C for 5 hours under N₂ protection. The resulting mixture was diluted with water and EtOAc and extracted three times with EtOAc. The organic layer was dried over anhydrous MgSO₄, filtered, and concentrated. Purification was performed on a silica gel column, eluting with 800%–300% PE / EA solution to separate 6-4 (75 mg).

[0199] Step 4: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-(2-(1-methyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-4-yl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 6)

[0200] Ethanol (6 mL) was added to 6-4 (75 mg, 0.16 mmol) and PtO2 (34 mg, 0.15 mmol), and the reaction was carried out at room temperature for 3 hours under a hydrogen atmosphere. After the reaction was complete, the mixture was filtered, and the residue was washed with a small amount of ethanol. The filtrate was concentrated and then dry-coated onto a silica gel column for chromatography, eluting with 400%–10% PE / EA solution to give compound 6 (35 mg).

[0201] 1 H NMR (300 MHz, Chloroform-d) δ 7.70 (t, J = 8.0 Hz, 1H), 7.46 (s,1H), 7.42 – 7.34 (m, 2H), 7.27 (d, J = 1.9 Hz, 1H), 4.51 (dd, J = 11.3, 2.1Hz, 1H), 4.29 – 4.17 (m, 1H), 3.86 (s, 3H), 3.75 (td, J = 11.5, 3.2 Hz, 1H), 3.55 (s, 3H), 3.52 – 3.42 (m, 1H), 3.13 (d, J = 0.7 Hz, 3H), 2.25 (d, J =21.1 Hz, 1H), 2.06 (d, J = 12.8 Hz, 3H).

[0202] MS m / z (ES-API+): 485.20 [M+H] +

[0203] Example 7: Synthesis of 8-(4-chloro-2-fluorophenyl)-3-methyl-1-(methyl-d3)-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholine)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 7)

[0204]

[0205] Step 1: Synthesis of 6-chloro-8-(4-chloro-2-fluorophenyl)-3-methyl-1-(methyl-d3)pyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (7-1)

[0206] Dissolve 1-6 (100 mg, 0.29 mmol) in anhydrous DMF (10 mL), add NaH (11 mg, 0.44 mmol) at 0 °C, and react at room temperature for 5 min. Then add CD3I (134 mg, 59.5 μL, 0.96 mmol), and react at room temperature for 3 h. After the reaction is complete, quench the reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, elute with 800%–400% PE / EA solution, and separate to obtain 7-1 (90 mg).

[0207] Step 2: Synthesis of 8-(4-chloro-2-fluorophenyl)-3-methyl-1-(methyl-d3)-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholine)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 7)

[0208] Dissolve 7-1 (90 mg, 0.25 mmol) in DMSO (10 mL), add DIPEA (0.5 mL) and 1-8 (40 mg, 0.24 mmol), and react at 80 °C for 3 hours. Quench the reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, elute with 800%–400% PE / EA solution, and separate to obtain compound 7 (24.5 mg).

[0209] 1 H NMR (300 MHz, Chloroform-d) δ 7.63 – 7.50 (m, 2H), 7.43 (s, 1H), 7.32 (dd, J = 8.3, 1.9 Hz, 1H), 7.23 (dd, J = 9.6, 1.8 Hz, 1H), 4.71 (d, J =13.3 Hz, 1H), 4.63 – 4.48 (m, 2H), 4.07 (d, J = 11.6 Hz, 1H), 3.90 (s, 3H), 3.76 (td, J = 10.2, 9.0, 3.5 Hz, 1H), 3.51 (s, 3H), 3.23 (ddd, J = 23.7,14.2, 11.3 Hz, 2H).

[0210] MS m / z (ES-API+): 489.20 [M+H] +

[0211] Example 8: Synthesis of 8-(2,4-difluorophenyl)-1,3-dimethyl-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholine)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 8)

[0212]

[0213] Step 1: Synthesis of 5-amino-2,6-dichloropyrimidine-4-carboxylic acid (8-1)

[0214] Add a solution of LiOH (2.8 g, 66 mmol) in H₂O (24 mL) to a THF (36 mL) solution of 1-1 (5.2 g, 22 mmol). Stir at room temperature for 2 hours. Dilute with water and adjust the pH to approximately 4 with 6 M HCl aqueous solution. Extract three times with EA. Dry the combined organic layers with anhydrous Na₂SO₄ and concentrate under vacuum to obtain crude product 8-1 (4.7 g).

[0215] Step 2: Synthesis of 5-amino-2,6-dichloro-N-methylpyrimidine-4-carboxamide (8-2)

[0216] 8-1 (4.7 g, 22.7 mmol), methylamine hydrochloride (1.52 g, 22.7 mmol), and HATU (13 g, 34 mmol) were stirred in DMF (60 mL) at 20°C under nitrogen atmosphere, and DIPEA (8.8 g, 68.4 mmol) was added. The reaction mixture was stirred at 50°C for 2 hours. The reaction mixture was diluted with H2O (200 mL) and extracted with EtOAc. The combined organic matter was dried over anhydrous MgSO4, filtered, and concentrated under vacuum to give the residue. The residue was purified by elution with 800%–300% PE / EA solution on a silica gel column to give product 8-2 (1.7 g).

[0217] Step 3: Synthesis of 5-amino-2-chloro-6-(2,4-difluorophenyl)-N-methylpyrimidine-4-carboxamide (8-4)

[0218] To a solution of 1,4-dioxane (1.53 g, 6.96 mmol) and 8-3 (1.1 g, 6.96 mmol) in water (50 mL: 5 mL), Pd(dppf)Cl2 (1 g, 1.39 mmol) and K3PO4 (4.4 g, 20.88 mmol) were added, and the mixture was stirred at 55 °C for 3 hours under N2 protection. The resulting mixture was diluted with water and EtOAc and extracted three times with EtOAc. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. Purification was performed on a silica gel column, eluting with 800%–300% PE / EA solution to give 8-4 (1.6 g).

[0219] Step 4: Synthesis of 6-chloro-8-(2,4-difluorophenyl)-3-methylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (8-5)

[0220] Dissolve 8-4 (1.6 g, 5.36 mmol) in 50 mL of anhydrous tetrahydrofuran solution. Add NaH (514 mg, 21.4 mmol) at 0 °C and react at room temperature for 5 min. Then add triphosgene (1.6 mg, 5.36 mmol) and react at room temperature for 10 h. After the reaction is complete, quench the reaction with saturated sodium bicarbonate solution, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, elute with 800%–200% PE / EA solution, and obtain 8-5 (1.09 mg).

[0221] Step 5: Synthesis of 6-chloro-8-(2,4-difluorophenyl)-1,3-dimethylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (8-6)

[0222] Dissolve 8-5 (1.09 mg, 3.36 mmol) in 50 mL of anhydrous DMF solution. Then add NaH (121 mg, 5.05 mmol) at 0 °C and react at room temperature for 20 min. Then add CH3I (525 mg, 3.70 mmol) and react at room temperature for 3 h. After the reaction is complete, quench the reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, elute with 800%–400% PE / EA solution to obtain 8-6 (645 mg).

[0223] 1 H NMR (300 MHz, DMSO-d6) δ 7.79 (td, J = 8.6, 6.5 Hz, 1H), 7.55 (ddd,J = 10.5, 9.3, 2.5 Hz, 1H), 7.36 (tdd, J = 8.5, 2.5, 0.9 Hz, 1H), 3.34 (s,3H), 2.91 (s,3H).

[0224] Step 6: Synthesis of 8-(2,4-difluorophenyl)-1,3-dimethyl-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholine)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 8)

[0225] Compound 8-6 (140 mg, 0.41 mmol), compound 1-8 (90 mg, 0.54 mmol), and DIPEA (267 mg, 2.07 mmol) were dissolved in DMSO (5 mL) and stirred at 80 °C for 4 hours. After the reaction was complete, the reaction was quenched with water, and the mixture was extracted three times with ethyl acetate. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. The mixture was purified on a silica gel column and eluted with 800%–400% PE / EA solution to obtain compound 8 (70 mg).

[0226] 1 H NMR (400 MHz, Chloroform-d) δ 7.65 (td, J = 8.4, 6.3 Hz, 1H), 7.55(d, J = 0.8 Hz, 1H), 7.45 (s, 1H), 7.09 (tdd, J = 8.7, 2.5, 0.9 Hz, 1H), 6.96(ddd, J = 9.9, 8.6, 2.4 Hz, 1H), 4.74 (d, J = 13.3 Hz, 1H), 4.66 – 4.52 (m,2H), 4.09 (ddd, J = 11.6, 3.5, 1.8 Hz, 1H), 3.92 (s, 3H), 3.78 (td, J = 11.5,2.8 Hz, 1H), 3.53 (s, 3H), 3.31 (q, J = 12.0, 10.5 Hz, 1H), 3.21 (dd, J =13.3, 10.3 Hz, 1H), 3.06 (s, 3H).

[0227] MS m / z (ES-API+): 470.20 [M+H] +

[0228] Example 9: Synthesis of 6-(2-(1-cyclopropyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-4-yl)-8-(2,4-difluorophenyl)-1,3-dimethylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 9)

[0229]

[0230] Step 1: Synthesis of 6-(6-(1-cyclopropyl-1H-pyrazol-4-yl)-3,6-dihydro-2H-pyran-4-yl)-8-(2,4-difluorophenyl)-1,3-dimethylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (9-1)

[0231] To solutions of 1,4-dioxane (280 mg, 0.83 mmol) and 2-3 (314 mg, 0.9 mmol) in water (10 mL: 2 mL), Pd(dppf)₂Cl₂·DCM (54 mg, 0.066 mmol) and K₂CO₃ (343 mg, 2.48 mmol) were added, and the mixture was stirred at 80 °C for 5 hours under N₂ protection. The resulting mixture was diluted with water and EtOAc and extracted three times with EtOAc. The organic layer was dried over anhydrous MgSO₄, filtered, and concentrated. Purification was performed on a silica gel column, eluting with 0%–300% PE / EA solution to give 9-1 (100 mg).

[0232] 1 H NMR (300 MHz, Chloroform-d) δ 7.79 (q, J = 7.9 Hz, 1H), 7.52 (s,1H), 7.50 (s, 1H), 7.39 (s, 1H), 7.14 (t, J = 7.4 Hz, 1H), 7.03 – 6.94 (m,1H), 5.42 (d, J = 2.8 Hz, 1H), 4.10 – 4.03 (m, 1H), 3.90 (ddd, J = 9.8, 6.2,3.5 Hz, 1H), 3.57 (s, 4H), 3.15 (s, 3H), 2.97 – 2.76 (m, 2H), 1.12 (s, 2H),1.01 (d, J = 7.4 Hz, 2H).

[0233] Step 2: Synthesis of 6-(2-(1-cyclopropyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-4-yl)-8-(2,4-difluorophenyl)-1,3-dimethylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 9)

[0234] Ethanol (5 mL) was added to 9-1 (100 mg, 0.20 mmol) and PtO2 (46 mg, 0.20 mmol), and the reaction was carried out at room temperature for 3 hours under a hydrogen atmosphere. After the reaction was completed, the mixture was filtered, and the residue was washed with a small amount of ethanol. The filtrate was concentrated and then dry-coated onto a silica gel column for chromatography, eluting with 400%–10% PE / EA solution to give compound 9 (30 mg).

[0235] 1H NMR (300 MHz, Chloroform-d) δ 7.78 (td, J = 8.4, 6.2 Hz, 1H), 7.48(s, 2H), 7.15 (td, J = 7.9, 2.1 Hz, 1H), 6.99 (ddd, J = 10.0, 8.5, 2.4 Hz,1H), 4.52 (dd, J = 11.3, 2.1 Hz, 1H), 4.30 – 4.17 (m, 1H), 3.77 (td, J =11.5, 3.3 Hz, 1H), 3.57 (s, 5H), 3.14 (s, 3H), 2.06 (s, 2H), 1.83 (s, 2H),1.10 (q, J = 4.4 Hz, 2H), 1.01 (dd, J = 7.2, 5.0 Hz, 2H).

[0236] MS m / z (ES-API+): 495.20 [M+H] +

[0237] Example 10: Synthesis of 8-(2,4-difluorophenyl)-1,3-dimethyl-6-(2-(1-methyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-4-yl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 10)

[0238]

[0239] Step 1: Synthesis of 8-(2,4-difluorophenyl)-1,3-dimethyl-6-(6-(1-methyl-1H-pyrazol-4-yl)-3,6-dihydro-2H-pyran-4-yl)pyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (10-1)

[0240] To solutions of 1,4-dioxane (225 mg, 0.66 mmol) and 6-3 (231 mg, 0.80 mmol) in water (10 mL: 2 mL), Pd(dppf)₂Cl₂·DCM (43 mg, 0.053 mmol) and K₂CO₃ (275 mg, 1.99 mmol) were added, and the mixture was stirred at 80 °C for 5 hours under N₂ protection. The resulting mixture was diluted with water and EtOAc and extracted three times with EtOAc. The organic layer was dried over anhydrous MgSO₄, filtered, and concentrated. Purification was performed on a silica gel column, eluting with 0%–300% PE / EA solution to give 10⁻¹ (10⁴ mg).

[0241] Step 2: Synthesis of 8-(2,4-difluorophenyl)-1,3-dimethyl-6-(2-(1-methyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-4-yl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 10)

[0242] Ethanol (5 mL) was added to 10⁻¹ (10⁴ mg, 0.22 mmol) and PtO₂ (51 mg, 0.22 mmol), and the mixture was reacted at room temperature for 3 hours under a hydrogen atmosphere. After the reaction was complete, the mixture was filtered, and the residue was rinsed with a small amount of ethanol. The filtrate was concentrated and then dry-coated onto a silica gel column for chromatography, eluting with 400%–10% PE / EA solution to give compound 10 (35 mg).

[0243] 1 H NMR (300 MHz, Chloroform-d) δ 7.77 (td, J = 8.4, 6.2 Hz, 1H), 7.48(s, 1H), 7.39 (s, 1H), 7.19 – 7.09 (m, 1H), 6.99 (ddd, J = 10.1, 8.5, 2.4 Hz, 1H), 4.53 (dd, J = 11.4, 2.2 Hz, 1H), 4.23 (ddd, J = 11.5, 4.2, 2.0 Hz, 1H), 3.88 (s, 3H), 3.77 (td, J = 11.5, 3.3 Hz, 1H), 3.57 (s, 3H), 3.55 – 3.46 (m,1H), 3.14 (s, 3H), 2.30 (s, 2H), 2.05 (s, 2H).

[0244] MS m / z (ES-API+): 549.20 [M+H] +

[0245] Example 11: Synthesis of 8-(2-fluoro-4-(trifluoromethyl)phenyl)-3-isopropyl-1-methyl-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholine)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 11)

[0246]

[0247] Step 1: Synthesis of ethyl 5-amino-2-chloro-6-(2-fluoro-4-(trifluoromethyl)phenyl)pyrimidine-4-carboxylate (11-2)

[0248] To a solution of 1,4-dioxane (4 g, 16.9 mmol) and 11-1 (3.52 g, 16.9 mmol) in water (50 mL: 5 mL), Pd(dppf)₂Cl₂ (2.5 g, 3.38 mmol) and K₃PO₄ (10.8 g, 50.7 mmol) were added, and the mixture was stirred at 55 °C for 2 hours under N₂ protection. The resulting mixture was diluted with water and EtOAc and extracted three times with EtOAc. The organic layer was dried over anhydrous MgSO₄, filtered, and concentrated. Purification was performed on a silica gel column, eluting with 800%–300% PE / EA solution to give 11-2 (4.4 g).

[0249] 1 H NMR (300 MHz, Chloroform-d) δ 7.73 – 7.66 (m, 1H), 7.62 (dd, J =8.1, 1.5 Hz, 1H), 7.53 (dd, J = 9.6, 1.5 Hz, 1H), 5.86 (s, 2H), 4.50 (q, J =7.1 Hz, 2H), 1.46 (t, J = 7.1 Hz, 3H).

[0250] Step 2: Synthesis of 5-amino-2-chloro-6-(2-fluoro-4-(trifluoromethyl)phenyl)pyrimidine-4-carboxylic acid (11-3)

[0251] A solution of LiOH·H₂O (1.52 g, 36.3 mmol) in H₂O (20 mL) was added to a THF (30 mL) solution of 11⁻² (4.4 g, 12.1 mmol). The mixture was stirred at room temperature for 2 hours. The reaction was quenched with water and 2 M HCl aqueous solution (30 mL), and the mixture was extracted three times with DCM. The combined organic layers were dried over anhydrous Na₂SO₄ and concentrated under vacuum to give crude product 11⁻³ (3.7 g).

[0252] Step 3: Synthesis of 5-amino-2-chloro-6-(2-fluoro-4-(trifluoromethyl)phenyl)-N-isopropylpyrimidine-4-carboxamide (11-4)

[0253] 11-3 (1 g, 2.98 mmol), N-isopropylamine hydrochloride (428 mg, 4.47 mmol), and HATU (1.7 g, 4.47 mmol) were stirred in DMF (30 mL) at 20°C under nitrogen, and DIPEA (1.56 mL, 8.94 mmol) was added. The reaction mixture was stirred at 50°C for 2 hours. The reaction mixture was diluted with H2O (100 mL) and extracted with EtOAc. The combined organic matter was dried over anhydrous MgSO4, filtered, and concentrated under vacuum to give the residue. The residue was purified by elution with 800%–300% PE / EA solution on a silica gel column to give 11-4 (1.0 g).

[0254] Step 4: Synthesis of 6-chloro-8-(2-fluoro-4-(trifluoromethyl)phenyl)-3-isopropylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (11-5)

[0255] Dissolve 11-4 (1.4 g, 3.72 mmol) in 50 mL of anhydrous tetrahydrofuran solution. Add NaH (350 mg, 14.88 mmol) at 0 °C and react at room temperature for 5 min. Then add triphosgene (1.1 g, 3.70 mmol) and react at room temperature for 6 h. After the reaction is complete, quench the reaction with saturated sodium bicarbonate solution, concentrate under reduced pressure, add ice water, filter under reduced pressure, and wash the filter cake several times with ice water to obtain 11-5 (760 mg).

[0256] Step 5: Synthesis of 6-chloro-8-(2-fluoro-4-(trifluoromethyl)phenyl)-3-isopropyl-1-methylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (11-6)

[0257] Dissolve 11-5 (100 mg, 0.25 mmol) in 50 mL of anhydrous DMF solution. Then add NaH (9 mg, 0.37 mmol) at 0 °C and react at room temperature for 20 min. Next, add CH3I (117 mg, 0.825 mmol) and react at room temperature for 3 h. After the reaction is complete, quench the reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter, and concentrate. Purify on a silica gel column, eluting with 800%–400% PE / EA solution to obtain 11-6 (11 mg).

[0258] Step 6: Synthesis of 8-(2-fluoro-4-(trifluoromethyl)phenyl)-3-isopropyl-1-methyl-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholine)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 11)

[0259] Compound 11 (34 mg, 0.08 mmol) was dissolved in 10 mL of DMSO, followed by the addition of 0.5 mL of DIPEA and 1-8 (78 mg, 0.19 mmol). The reaction was carried out at 80 °C for 3 hours. The reaction was quenched with water, and the mixture was extracted three times with ethyl acetate. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. The mixture was purified on a silica gel column and eluted with 800%–400% PE / EA solution to obtain compound 11 (11 mg).

[0260] 1 H NMR (300 MHz, Chloroform-d) δ 7.76 (t, J = 7.5 Hz, 1H), 7.59 (d, J= 8.1 Hz, 1H), 7.55 – 7.37 (m, 3H), 5.23 (p, J = 6.9 Hz, 1H), 4.71 (d, J =13.3 Hz, 1H), 4.54 (d, J = 8.4 Hz, 2H), 4.07 (d, J = 12.0 Hz, 1H), 3.89 (s,3H), 3.76 (td, J = 10.6, 10.1, 4.8 Hz, 1H), 3.22 (dt, J = 24.2, 11.8 Hz, 2H),2.99 (s, 3H), 1.55 (d, J = 6.9 Hz, 6H).

[0261] MS m / z (ES-API+): 548.20 [M+H] +

[0262] Example 12: Synthesis of 3-cyclopropyl-8-(2-fluoro-4-(trifluoromethyl)phenyl)-1-methyl-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholine)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 12)

[0263]

[0264] Step 1: Synthesis of 5-amino-2-chloro-N-cyclopropyl-6-(2-fluoro-4-(trifluoromethyl)phenyl)pyrimidine-4-carboxamide (12-1)

[0265] 11-3 (1 g, 2.98 mmol), cyclopropylamine hydrochloride (428 mg, 4.47 mmol), and HATU (1.7 g, 4.47 mmol) were stirred in DMF (30 mL) at 20°C under nitrogen, and DIPEA (1.56 mL, 8.94 mmol) was added. The reaction mixture was stirred at 50°C for 2 hours. The reaction mixture was diluted with H2O (100 mL) and extracted with EtOAc. The combined organic matter was dried over anhydrous MgSO4, filtered, and concentrated under vacuum to give the residue. The residue was purified by elution of 800%–300% PE / EA solution onto a silica gel column to give 12-1 (1.0 g).

[0266] Step 2: Synthesis of 6-chloro-3-cyclopropyl-8-(2-fluoro-4-(trifluoromethyl)phenyl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (12-2)

[0267] Dissolve 12-1 (1.4 g, 3.72 mmol) in 50 mL of anhydrous tetrahydrofuran solution. Add NaH (350 mg, 14.88 mmol) at 0 °C and react at room temperature for 5 min. Then add triphosgene (1.1 g, 3.70 mmol) and react at room temperature for 6 h. After the reaction is complete, quench the reaction with saturated sodium bicarbonate aqueous solution, concentrate under reduced pressure, add ice water, filter under reduced pressure, and wash the filter cake several times with ice water to obtain 12-2 (760 mg).

[0268] Step 3: Synthesis of 6-chloro-3-cyclopropyl-8-(2-fluoro-4-(trifluoromethyl)phenyl)-1-methylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (12-3)

[0269] Dissolve 12-2 (100 mg, 0.25 mmol) in 50 mL of anhydrous DMF solution. Then add NaH (9 mg, 0.37 mmol) at 0 °C and react at room temperature for 20 min. Next, add CH3I (117 mg, 0.825 mmol) and react at room temperature for 3 h. After the reaction is complete, quench the reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter, and concentrate. Purify on a silica gel column, eluting with 800%–400% PE / EA solution to obtain 12-3 (11 mg).

[0270] Step 4: Synthesis of 3-cyclopropyl-8-(2-fluoro-4-(trifluoromethyl)phenyl)-1-methyl-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholine)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 12)

[0271] Dissolve compound 12-3 (34 mg, 0.08 mmol) in DMSO (10 mL). Add DIPEA (0.5 mL) and compound 1-8 (78 mg, 0.19 mmol), and react at 80 °C for 3 hours. Quench the reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, elute with 800%–400% PE / EA solution, and separate to obtain compound 12 (11 mg).

[0272] 1 H NMR (400 MHz, Chloroform-d) δ 7.77 (t, J = 7.5 Hz, 1H), 7.59 (d, J= 8.1 Hz, 1H), 7.55 – 7.34 (m, 3H), 4.85 – 4.67 (m, 1H), 4.56 (t, J = 11.6Hz, 2H), 4.07 (d, J = 11.3 Hz, 1H), 3.89 (s, 3H), 3.83 – 3.69 (m, 1H), 3.23 (dt, J = 35.3, 11.7 Hz, 2H), 2.99 (s, 3H), 1.23 (dd, J = 15.9, 7.3 Hz, 3H),0.84 (d, J = 4.4 Hz, 2H).

[0273] MS m / z (ES-API+): 546.20 [M+H] +

[0274] Example 13: Synthesis of 8-(2-fluoro-4-(trifluoromethyl)phenyl)-1-methyl-3-(methyl-d3)-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholino)pyrimidinyl[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 13)

[0275]

[0276] Step 1: Synthesis of 5-amino-2-chloro-6-(2-fluoro-4-(trifluoromethyl)phenyl)-N-(methyl-d3)pyrimidine-4-carboxamide (13-1)

[0277] 11-3 (1 g, 2.98 mmol), deuterated methylamine hydrochloride (316 mg, 4.47 mmol), and HATU (1.7 g, 4.47 mmol) were stirred in DMF (30 mL) at 20°C under nitrogen, and DIPEA (1.56 mL, 8.94 mmol) was added. The reaction mixture was stirred at 50°C for 2 hours. The reaction mixture was diluted with H2O (100 mL) and extracted with EtOAc. The combined organic matter was dried over anhydrous MgSO4, filtered, and concentrated under vacuum to give the residue. The residue was purified by elution with PE / EA solution at 800%–300% on a silica gel column to give 13-1 (1.0 g).

[0278] Step 2: Synthesis of 6-chloro-8-(2-fluoro-4-(trifluoromethyl)phenyl)-3-(methyl-d3)pyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (13-2)

[0279] Dissolve 13-1 (1.0 g, 2.84 mmol) in 50 mL of anhydrous tetrahydrofuran solution. Add NaH (282 mg, 11.75 mmol) at 0 °C and react at room temperature for 5 min. Then add triphosgene (873 mg, 2.94 mmol) and react at room temperature for 6 h. After the reaction is complete, quench the crude reaction with saturated sodium bicarbonate aqueous solution, concentrate under reduced pressure, add ice water, filter under reduced pressure, and wash the filter cake several times with ice water to obtain 13-2 (900 mg).

[0280] Step 3: Synthesis of 6-chloro-8-(2-fluoro-4-(trifluoromethyl)phenyl)-1-methyl-3-(methyl-d3)pyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (13-3)

[0281] Dissolve 13-2 (100 mg, 0.25 mmol) in 50 mL of anhydrous DMF solution. Then add NaH (10 mg, 0.37 mmol) at 0 °C and react at room temperature for 20 min. Then add CH3I (117 mg, 0.825 mmol) and react at room temperature for 3 h. After the reaction is complete, quench the crude reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, elute with 800%–400% PE / EA solution, and obtain 13-3 (45 mg).

[0282] Step 4: Synthesis of 8-(2-fluoro-4-(trifluoromethyl)phenyl)-1-methyl-3-(methyl-d3)-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholino)pyrimidinyl[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 13)

[0283] Dissolve compound 13-3 (45 mg, 0.12 mmol) in DMSO (10 mL). Add DIPEA (0.5 mL) and compound 1-8 (28 mg, 0.16 mmol), and react at 80 °C for 3 hours. Quench the reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 800%–400% PE / EA solution to obtain compound 13 (38 mg).

[0284] 1 H NMR (300 MHz, Chloroform-d) δ 7.77 (t, J = 7.4 Hz, 1H), 7.60 (dd,J = 8.2, 1.5 Hz, 1H), 7.56 – 7.37 (m, 3H), 4.71 (d, J = 13.4 Hz, 1H), 4.64 –4.46 (m, 2H), 4.15 – 4.01 (m, 1H), 3.88 (s, 3H), 3.82 – 3.69 (m, 1H), 3.37 –3.12 (m, 2H), 3.02 (s, 3H).

[0285] MS m / z (ES-API+): 523.20 [M+H] +

[0286] Example 14: Synthesis of 8-(2-fluoro-4-(trifluoromethyl)phenyl)-1,3-dimethyl-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholine)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 14)

[0287]

[0288] Step 1: Synthesis of 5-amino-2-chloro-6-(2-fluoro-4-(trifluoromethyl)phenyl)-N-methylpyrimidine-4-carboxamide (14-1)

[0289] 11-3 (3 g, 8.94 mmol), methylamine hydrochloride (905 mg, 13.41 mmol), and HATU (5 g, 13.41 mmol) were stirred in DMF (30 mL) at 20°C under nitrogen. DIPEA (3.46 g, 26.82 mmol) was then added. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was diluted with H2O (100 mL) and extracted with EtOAc. The combined organic matter was dried over anhydrous MgSO4, filtered, and concentrated under vacuum to obtain the residue. The residue was purified by elution with 800%–300% PE / EA solution on a silica gel column to give 14-1 (3.1 g).

[0290] Step 2: Synthesis of 6-chloro-8-(2-fluoro-4-(trifluoromethyl)phenyl)-3-methylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (14-2)

[0291] Dissolve 14-1 (3.1 g, 8.91 mmol) in 30 mL of anhydrous tetrahydrofuran solution. Add NaH (850 mg, 35.63 mmol) at 0 °C and react at room temperature for 5 min. Then add triphosgene (2.64 g, 8.91 mmol) in anhydrous tetrahydrofuran (10 mL) and react at room temperature for 10 h. After the reaction is complete, quench the reaction with saturated sodium bicarbonate aqueous solution, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, elute with 800%–200% PE / EA solution, and separate to obtain 14-2 (2.5 g).

[0292] Step 3: Synthesis of 6-chloro-8-(2-fluoro-4-(trifluoromethyl)phenyl)-1,3-dimethylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (14-3)

[0293] Dissolve 14-2 (2.5 g, 6.68 mmol) in 15 mL of anhydrous DMF solution. Then add NaH (240 mg, 10.0 mmol) at 0 °C and react at room temperature for 20 min. Next, add CH3I (1.04 g, 7.35 mmol) and react at room temperature for 3 h. After the reaction is complete, quench the reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 800%–400% PE / EA solution to obtain 14-3 (2.5 g).

[0294] Step 4: Synthesis of 8-(2-fluoro-4-(trifluoromethyl)phenyl)-1,3-dimethyl-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholine)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 14)

[0295] Dissolve compound 14-3 (96 mg, 0.26 mmol) in 3 mL of DMSO solution. Add DIPEA (103 mg, 0.76 mmol) and 1-8 (45 mg, 0.26 mmol), and react at 80 °C for 3 h. Quench the reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 800%–400% PE / EA solution to obtain compound 14 (50 mg).

[0296] 1 H NMR (300 MHz, Chloroform-d) δ 7.77 (t, J = 7.5 Hz, 1H), 7.60 (d, J= 8.0 Hz, 1H), 7.53 (s, 1H), 7.50 – 7.45 (m, 1H), 7.44 (s, 1H), 4.71 (d, J =13.3 Hz, 1H), 4.62 – 4.50 (m, 2H), 4.07 (dd, J = 9.5, 3.0 Hz, 1H), 3.90 (s,3H), 3.78 – 3.74 (m, 1H), 3.51 (s, 3H), 3.23 (dt, J = 23.8, 11.9 Hz, 2H),3.02 (s, 3H).

[0297] MS m / z (ES-API+): 520.20 [M+H] +

[0298] Example 15: Synthesis of 3-ethyl-8-(2-fluoro-4-(trifluoromethyl)phenyl)-1-methyl-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholine)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 15)

[0299]

[0300] Step 1: Synthesis of 5-amino-2-chloro-N-ethyl-6-(2-fluoro-4-(trifluoromethyl)phenyl)pyrimidine-4-carboxamide (15-1)

[0301] 11-3 (1 g, 2.99 mmol), ethylamine hydrochloride (365 mg, 4.48 mmol), and HATU (1.7 g, 4.48 mmol) were stirred in DMF (30 mL) under nitrogen at 20°C, and DIPEA (1.16 g, 8.96 mmol) was added. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was diluted with H2O (100 mL) and extracted with EtOAc. The combined organic matter was dried over anhydrous MgSO4, filtered, and concentrated under vacuum to give the residue. The residue was purified by elution with 800%–300% PE / EA solution on a silica gel column to give 15-1 (1.12 g).

[0302] Step 2: Synthesis of 6-chloro-3-ethyl-8-(2-fluoro-4-(trifluoromethyl)phenyl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (15-2)

[0303] Dissolve 15-1 (1.12 g, 3.10 mmol) in 30 mL of anhydrous tetrahydrofuran solution. Add NaH (298 mg, 12.40 mmol) at 0 °C and react at room temperature for 5 min. Then add triphosgene (920 mg, 3.10 mmol) in 10 mL of anhydrous tetrahydrofuran and react at room temperature for 10 h. After the reaction is complete, quench the reaction with saturated sodium bicarbonate aqueous solution, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, elute with 800%–200% PE / EA solution, and separate to obtain 15-2 (760 mg).

[0304] Step 3: Synthesis of 6-chloro-3-ethyl-8-(2-fluoro-4-(trifluoromethyl)phenyl)-1-methylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (15-3)

[0305] Dissolve 15-2 (100 mg, 0.26 mmol) in 5 mL of anhydrous DMF solution. Then add NaH (10 mg, 0.39 mmol) at 0 °C and react at room temperature for 20 min. Next, add CH3I (40 mg, 0.28 mmol) and react at room temperature for 3 h. After the reaction is complete, quench the reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 800%–400% PE / EA solution to obtain 15-3 (50 mg).

[0306] Step 4: Synthesis of 3-ethyl-8-(2-fluoro-4-(trifluoromethyl)phenyl)-1-methyl-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholine)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 15)

[0307] Dissolve 15-3 (50 mg, 0.12 mmol) in 3 mL of DMSO solution. Add DIPEA (80 mg, 0.62 mmol) and 1-8 (27 mg, 0.16 mmol), and react at 80 °C for 3 hours. Quench the reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 800%–400% PE / EA solution to obtain compound 15 (41 mg).

[0308] 1 H NMR (300 MHz, Chloroform-d) δ 7.76 (t, J = 7.5 Hz, 1H), 7.64 –7.56 (m, 1H), 7.55 – 7.39 (m, 3H), 4.71 (d, J = 13.5 Hz, 1H), 4.56 (t, J =10.2 Hz, 2H), 4.16 (q, J = 7.0 Hz, 2H), 4.07 (d, J = 11.3 Hz, 1H), 3.89 (s,3H), 3.82 – 3.70 (m, 1H), 3.35 – 3.12 (m, 2H), 3.02 (s, 3H), 1.32 (t, J = 7.0Hz, 3H).

[0309] MS m / z (ES-API+): 534.20 [M+H] +

[0310] Example 16: Synthesis of 8-(2,4-difluorophenyl)-1,3-dimethyl-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholine)pyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (compound 16)

[0311]

[0312] Step 1: Synthesis of 3-amino-2,6-dichloro-N-methylisonicotinamide (16-2)

[0313] 16-1 (4 g, 19.4 mmol) was added to DMF (40 mL), along with HATU (11.1 g, 29.1 mmol), DIPEA (5.1 mL, 58.2 mmol), and methylamine hydrochloride (1.97 g, 29.1 mmol). The reaction mixture was allowed to react overnight at room temperature. The reaction mixture was then poured into 200 mL of saturated saline solution, stirred, and filtered to obtain 16-2 (4 g).

[0314] 1 H NMR (400 MHz, Chloroform-d) δ 7.18 (s, 1H), 6.25 (s, 1H), 5.96 (s,2H), 3.02 (d, J = 4.8 Hz, 3H).

[0315] Step 2: Synthesis of 6,8-dichloro-3-methylpyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (16-3)

[0316] 16-2 (4.4 g, 20 mmol) was added to anhydrous THF (8 mL), and NaH was added under an ice-water bath. After stirring for one hour, a THF solution of triphosgene (201 mg, 0.68 mmol) was added dropwise, and the reaction was allowed to proceed overnight. Column chromatography yielded 16-3 (3.5 g).

[0317] 1 H NMR (300 MHz, DMSO-d6) δ 11.85 (s, 1H), 8.17 (s, 1H), 3.56 (s, 3H).

[0318] Step 3: Synthesis of 6-chloro-8-(2,4-difluorophenyl)-3-methylpyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (16-4)

[0319] To a solution of 1,4-dioxane (1.4 g, 5.71 mmol) and 8-3 (900 mg, 5.73 mmol) in water (20 mL: 5 mL), Pd(dppf)Cl2 (836 mg, 1.14 mmol) and K3PO4 (3.6 g, 17 mmol) were added, and the mixture was stirred at 55 °C for 2 hours under N2 protection. The resulting mixture was diluted with water and EtOAc and extracted three times with EtOAc. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. Purification was performed on a silica gel column, eluting with 0%–300% PE / EA solution to give 16-4 (1 g).

[0320] 1H NMR (300 MHz, DMSO-d6) δ 11.23 (s, 1H), 7.92 (d, J = 1.2 Hz, 1H), 7.57 (td, J = 8.6, 6.6 Hz, 1H), 7.43 (ddd, J = 10.5, 9.4, 2.4 Hz, 1H), 7.31 –7.19 (m, 1H), 3.27 (s, 3H).

[0321] Step 4: Synthesis of 6-chloro-8-(2,4-difluorophenyl)-1,3-dimethylpyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (16-5)

[0322] NaH (148 mg, 6.2 mmol) was added to a dry DMF solution of 16-4 (1 g, 3.1 mmol) and stirred for 5 minutes under an ice-water bath. CH3I (483 mg, 3.4 mmol) was then added, and the reaction was carried out at room temperature for 2 hours. Water was added, and a solid precipitated. The solid was then filtered to obtain 16-5 (500 mg).

[0323] 1 H NMR (300 MHz, Chloroform-d) δ 8.08 (s, 1H), 7.66 (td, J = 8.4, 6.1Hz, 1H), 7.08 (td, J = 8.2, 2.5 Hz, 1H), 6.98 – 6.87 (m, 1H), 3.51 (s, 3H),3.11 (s, 3H).

[0324] Step 5: Synthesis of 8-(2,4-difluorophenyl)-1,3-dimethyl-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholine)pyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (compound 16)

[0325] 16-5 (100 mg, 0.3 mmol), 1-8 (59 mg, 0.35 mmol), Xphos PdG4 (25.5 mg, 0.03 mmol), and t-BuONa (28 mg, 0.29 mmol) were added to a two-necked flask. Anhydrous 1,4-dioxane (5 mL) was added under N2 protection, and the reaction was carried out at 100 °C for 5 hours. The reaction was quenched with water, extracted three times with EtOAc, and the combined organic layers were dried over anhydrous Na2SO4 and concentrated under vacuum. The mixture was purified on a silica gel column and eluted with PE:EA = 1:5 solution to give compound 16 (50 mg).

[0326] 1H NMR (300 MHz, Chloroform-d) δ 7.59 (td, J = 8.4, 6.3 Hz, 1H), 7.54(s, 1H), 7.42 (d, J = 14.5 Hz, 2H), 7.04 (td, J = 8.2, 2.5 Hz, 1H), 6.92 (td,J = 9.3, 2.5 Hz, 1H), 4.63 (d, J = 10.1 Hz, 1H), 4.26 (t, J = 11.9 Hz, 1H), 4.18 – 3.98 (m, 2H), 3.91 (s, 3H), 3.84 (t, J = 10.7 Hz, 1H), 3.50 (s, 3H),3.20 – 3.10 (m, 1H), 3.07 (s, 3H), 3.03 (d, J = 10.6 Hz, 1H).

[0327] MS m / z (ES-API+): 469.20 [M+H] +

[0328] Example 17: Synthesis of 8-(2,4-difluorophenyl)-1,3-dimethyl-6-(2-(1-methyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-4-yl)pyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (compound 17)

[0329]

[0330] Step 1: Synthesis of 8-(2,4-difluorophenyl)-1,3-dimethyl-6-(6-(1-methyl-1H-pyrazol-4-yl)-3,6-dihydro-2H-pyran-4-yl)pyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (17-1)

[0331] Pd(dppf)Cl2 (65 mg, 0.09 mmol) and K3PO4 (290 mg, 1.35 mmol) were added to a solution of 1,4-dioxane (150 mg, 0.45 mmol) and 6-3 (250 mg, 0.88 mmol) in water (4 mL: 1 mL). The mixture was reacted at 80 °C for 2 hours under N2 protection. The reaction was quenched with water, and the mixture was extracted three times with EtOAc. The combined organic layers were dried over anhydrous Na2SO4 and concentrated under vacuum. The mixture was purified on a silica gel column and eluted with PE:EA = 1:1 solution to give 17-1 (150 mg).

[0332] Step 2: Synthesis of 8-(2,4-difluorophenyl)-1,3-dimethyl-6-(2-(1-methyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-4-yl)pyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (compound 17)

[0333] Add 50 mg of PtO2 to 5 ml of an ethanol solution of 17-1 (100 mg, 0.21 mmol) and stir for 1.5 hours under a hydrogen atmosphere. Dry the organic layer and concentrate it under vacuum. Purify it on a silica gel column and elute with PE:EA = 1:1 solution to give compound 17 (30 mg).

[0334] 1 H NMR (300 MHz, Chloroform-d) δ 7.96 (s, 1H), 7.65 (tdd, J = 8.2,6.2, 1.4 Hz, 1H), 7.48 (s, 1H), 7.39 (s, 1H), 7.14 – 7.03 (m, 1H), 6.94 (ddd,J = 9.9, 8.6, 2.5 Hz, 1H), 4.54 (dd, J = 11.3, 2.0 Hz, 1H), 4.25 (dt, J =10.3, 2.9 Hz, 1H), 3.89 (d, J = 1.4 Hz, 3H), 3.85 – 3.71 (m, 1H), 3.52 (s,3H), 3.31 – 3.18 (m, 1H), 3.13 (s, 3H), 2.25 (d, J = 12.8 Hz, 1H), 2.07 –1.88 (m, 3H).

[0335] MS m / z (ES-API+): 468.20 [M+H] +

[0336] Example 18: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholine)pyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (compound 18)

[0337]

[0338] Step 1: Synthesis of 6-chloro-8-(4-chloro-2-fluorophenyl)-3-methylpyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (18-1)

[0339] To a solution of 1,4-dioxane (1 g, 4.1 mmol) and 1-2 (848 mg, 4.1 mmol) in water (20 mL: 5 mL), Pd(dppf)Cl2 (600 mg, 0.82 mmol) and K3PO4 (2.6 g, 12.3 mmol) were added, and the mixture was stirred at 60 °C for 2 hours under N2 protection. The resulting mixture was diluted with water and EtOAc and extracted three times with EtOAc. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. Purification was performed on a silica gel column, eluting with 0%–300% PE / EA solution to give 18-1 (0.86 g).

[0340] Step 2: Synthesis of 6-chloro-8-(4-chloro-2-fluorophenyl)-1,3-dimethylpyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (18-2)

[0341] NaH (83 mg, 3.5 mmol) was added to anhydrous DMF solution of 18-1 (0.86 g, 2.3 mmol) and stirred for 5 minutes under an ice-water bath. CH3I (360 mg, 2.5 mmol) was then added, and the reaction was carried out at room temperature for 3 hours. Water was added, and a solid precipitated. The solid was then filtered to obtain 18-2 (870 mg).

[0342] 1 H NMR (300 MHz, Chloroform-d) δ 8.10 (s, 1H), 7.64 (t, J = 8.0 Hz,1H), 7.35 (dd, J = 8.3, 1.9 Hz, 1H), 7.27 – 7.18 (m, 1H), 3.54 (d, J = 8.2Hz, 3H), 3.14 (s, 3H).

[0343] Step 3: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholine)pyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (compound 18)

[0344] 18-2 (100 mg, 0.3 mmol), 1-8 (59 mg, 0.35 mmol, 1.2 eq), Xphos Pd G4 (25.5 mg, 0.03 mmol), and t-BuONa (28 mg, 0.29 mmol) were added to a two-necked flask. Anhydrous 1,4-dioxane (5 mL) was added under N2 protection, and the reaction was carried out at 100 °C for 5 hours. The reaction was quenched with water, extracted three times with EtOAc, and the combined organic layers were dried over anhydrous Na2SO4 and concentrated under vacuum. The mixture was purified on a silica gel column and eluted with PE:EA = 1:5 solution to give compound 18 (50 mg).

[0345] 1 H NMR (300 MHz, Chloroform-d) δ 7.57 – 7.50 (m, 2H), 7.42 (s, 1H), 7.38 (s, 1H), 7.30 – 7.25 (m, 1H), 7.18 (dd, J = 9.5, 1.9 Hz, 1H), 4.60 (d, J= 10.3 Hz, 1H), 4.23 (t, J = 12.2 Hz, 1H), 3.99 (d, J = 3.9 Hz, 1H), 3.90 (d,J = 2.5 Hz, 3H), 3.82 (t, J = 11.1 Hz, 1H), 3.76 – 3.69 (m, 1H), 3.48 (s, 3H), 3.13 (d, J = 10.5 Hz, 1H), 3.06 (s, 3H), 3.00 (d, J = 11.9 Hz, 1H).

[0346] MS m / z (ES-API+): 485.10 [M+H] +

[0347] Example 19: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-(2-(1-methyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-4-yl)pyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (compound 19)

[0348]

[0349] Step 1: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-(6-(1-methyl-1H-pyrazol-4-yl)-3,6-dihydro-2H-pyran-4-yl)pyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (19-1)

[0350] Pd(dppf)Cl2 (61 mg, 0.08 mmol) and K3PO4 (270 mg, 1.26 mmol) were added to a solution of 1,4-dioxane (150 mg, 0.42 mmol) and 6-3 (240 mg, 0.84 mmol) in water (4 mL: 1 mL). The reaction was carried out at 80 °C for 2 hours under N2 protection. The reaction was quenched with water, extracted three times with EtOAc, and the combined organic layers were dried over anhydrous Na2SO4 and concentrated under vacuum. The mixture was purified on a silica gel column and eluted with PE:EA = 1:1 solution to give 19-1 (150 mg).

[0351] Step 2: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-(2-(1-methyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-4-yl)pyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (compound 19)

[0352] Add 50 mg of PtO2 to 5 ml of an ethanol solution of 19-1 (100 mg, 0.21 mmol) and stir for 1.5 hours under a hydrogen atmosphere. Dry the organic layer and concentrate it under vacuum. Purify it on a silica gel column and elute with PE:EA = 1:1 solution to give compound 19 (30 mg).

[0353] MS m / z (ES-API+): 484.10 [M+H] +

[0354] Example 20: Synthesis of 8-(2-fluoro-4-(trifluoromethyl)phenyl)-1,3-dimethyl-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholine)pyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (compound 20)

[0355]

[0356] Step 1: Synthesis of 6-chloro-8-(2-fluoro-4-(trifluoromethyl)phenyl)-3-methylpyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (20-1)

[0357] To a solution of 1,4-dioxane (1.5 g, 6.1 mmol) and 11-1 (1.07 g, 6.1 mmol) in water (20 mL: 5 mL), Pd(dppf)Cl2 (895 mg, 1.22 mmol) and K3PO4 (3.8 g, 18 mmol) were added, and the mixture was stirred at 55 °C for 2 hours under N2 protection. The resulting mixture was diluted with water and EtOAc and extracted three times with EtOAc. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. Purification was performed on a silica gel column, eluting with 0%–300% PE / EA solution to give 20-1 (1.4 g).

[0358] 1 H NMR (300 MHz, Chloroform-d) δ 8.97 (s, 1H), 8.10 (d, J = 0.6 Hz,1H), 7.77 (t, J = 7.5 Hz, 1H), 7.68 (d, J = 8.8 Hz, 1H), 7.57 (d, J = 9.9 Hz,1H), 3.29 (s, 3H).

[0359] Step 2: Synthesis of 6-chloro-8-(2-fluoro-4-(trifluoromethyl)phenyl)-1,3-dimethylpyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (20-2)

[0360] NaH (198 mg, 8.25 mmol) was added to anhydrous DMF solution of 20⁻¹ (1.4 g, 4.1 mmol) and stirred for 5 minutes under ice-water bath conditions. CH₃I (645 mg, 4.54 mmol) was then added, and the reaction was carried out at room temperature for 2 hours. Water was added, and a solid precipitated. The solid was then filtered to obtain 20⁻² (880 mg).

[0361] 1 H NMR (300 MHz, Chloroform-d) δ 8.15 (s, 1H), 7.85 (t, J = 7.5 Hz,1H), 7.64 (dd, J = 7.9, 1.4 Hz, 1H), 7.48 (dd, J = 9.4, 1.6 Hz, 1H), 3.54 (s,3H), 3.13 (d, J = 0.7 Hz, 3H).

[0362] Step 3: Synthesis of 8-(2-fluoro-4-(trifluoromethyl)phenyl)-1,3-dimethyl-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholine)pyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (compound 20)

[0363] 20-2 (150 mg, 0.42 mmol), 1-8 (59 mg, 0.51 mmol), XphosPd G4 (36 mg, 0.04 mmol), and t-BuONa (41 mg, 0.42 mmol) were added to a two-necked flask. Anhydrous 1,4-dioxane (5 mL) was added under N2 protection, and the reaction was carried out at 100 °C for 5 hours. The reaction was quenched with water, extracted three times with EtOAc, and the combined organic layers were dried over anhydrous Na2SO4 and concentrated under vacuum. The mixture was purified on a silica gel column and eluted with PE:EA = 1:5 solution to give compound 20 (50 mg).

[0364] 1 H NMR (300 MHz, Chloroform-d) δ 7.77 (t, J = 7.5 Hz, 1H), 7.58 (dd,J = 8.0, 1.6 Hz, 1H), 7.55 (d, J = 0.7 Hz, 1H), 7.49 – 7.41 (m, 3H), 4.63 (d,J = 10.3 Hz, 1H), 4.26 (t, J = 12.3 Hz, 1H), 4.17 – 4.09 (m, 1H), 4.05 (d, J= 14.9 Hz, 1H), 3.92 (s, 3H), 3.85 (t, J = 11.5 Hz, 1H), 3.51 (s, 3H), 3.23 –2.97 (m, 5H).

[0365] MS m / z (ES-API+): 519.20 [M+H] +

[0366] Example 21: Synthesis of 8-(2-fluoro-4-(trifluoromethyl)phenyl)-1,3-dimethyl-6-(2-(1-methyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-4-yl)-6,7-dihydropyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 21)

[0367]

[0368] Step 1: Synthesis of 8-(2-fluoro-4-(trifluoromethyl)phenyl)-1,3-dimethyl-6-(6-(1-methyl-1H-pyrazol-4-yl)-3,6-dihydro-2H-pyran-4-yl)pyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (21-1)

[0369] To a mixed solution of 14-3 (150 mg, 0.38 mmol) and 6-3 (134 mg, 0.46 mmol) in 8 mL of 1,4-dioxane and 2 mL of H2O, Pd(dppf)2Cl2·DCM (26 mg, 0.03 mmol) and KOAc (161 mg, 1.16 mmol) were added. The mixture was stirred at 80 °C for 5 hours under N2 protection. The resulting mixture was diluted with water, extracted three times with EA, the organic layer was dried over MgSO4, filtered and concentrated, and purified by silica gel column chromatography (PE / EA = 800%–300%) to obtain 21-1 (90 mg).

[0370] Step 2: Synthesis of 8-(2-fluoro-4-(trifluoromethyl)phenyl)-1,3-dimethyl-6-(2-(1-methyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-4-yl)-6,7-dihydropyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (21-2)

[0371] Ethanol (5 mL) was added to 21-1 (90 mg, 0.17 mmol) and PtO2 (37 mg, 0.16 mmol), and the mixture was reacted at room temperature for 3 hours under a hydrogen atmosphere. After the reaction was completed, the mixture was filtered, and the residue was washed with a small amount of ethanol. The filtrate was concentrated and purified by silica gel column chromatography (PE / EA = 400% - 10%) to obtain 21-2 (36 mg).

[0372] Step 3: Synthesis of 8-(2-fluoro-4-(trifluoromethyl)phenyl)-1,3-dimethyl-6-(2-(1-methyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-4-yl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 21)

[0373] Compound 21-2 (30 mg, 0.06 mmol) and MnO2 (200 mg, 2.3 mmol) were dissolved in DCE (5 mL) and reacted at room temperature for 3 hours. After the reaction was complete, the mixture was filtered, and the residue was washed with a small amount of DCE. The filtrate was concentrated and purified by silica gel column chromatography (PE / EA = 400% - 10%) to obtain compound 21 (5.17 mg).

[0374] 1H NMR (300 MHz, Chloroform-d) δ 8.20 (t, J = 7.5 Hz, 1H), 7.96 (d, J= 8.2 Hz, 1H), 7.76 (s, 1H), 7.66 (d, J = 6.8 Hz, 2H), 4.82 (d, J = 11.4 Hz,1H), 4.53 (d, J = 11.8 Hz, 1H), 4.25 (d, J = 13.6 Hz, 1H), 4.16 (s, 3H), 3.87(s, 3H), 3.80 (d, J = 11.5 Hz, 1H), 3.42 (s, 3H), 1.66 – 1.46 (m, 4H).

[0375] MS m / z (ES-API+): 519.20 [M+H] +

[0376] Example 22: Synthesis of 8-(4-chloro-2-fluorophenyl)-6-(4,4-difluoro-3-(2-methylpyridin-4-yl)piperidin-1-yl)-1,3-dimethylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 22)

[0377]

[0378] Step 1: Synthesis of tert-butyl 3-(2-methylpyridin-4-yl)-4-oxoperpiperidine-1-carboxylic acid (22-2)

[0379] Under a nitrogen atmosphere, 22-1 (5.0 g, 25.1 mmol), 4-bromo-2-methylpyridine (4.317 g, 25.1 mmol), palladium acetate (563 mg, 2.51 mmol), tri-tert-butylphosphine tetrafluoroborate (728 mg, 2.51 mmol), and sodium tert-butoxide (2628 mg, 27.3 mmol) were dissolved in anhydrous tetrahydrofuran (100 ml). The reaction mixture was stirred at 60ºC for 12 hours. After the reaction was completed, the reaction mixture was concentrated and purified by column chromatography (PE:EA = 1:1) to obtain 22-2 (1.5 g).

[0380] 1H NMR (300 MHz, Chloroform-d) δ 8.48 (d, J = 5.4 Hz, 1H), 7.12 –6.91 (m, 2H), 4.12 – 4.05 (m, 1H), 3.57 (dddd, J = 22.2, 13.8, 10.0, 5.1 Hz,4H), 2.79 – 2.66 (m, 2H), 2.57 (s, 3H), 1.53 – 1.51 (m, 9H).

[0381] Step 2: Synthesis of tert-butyl 4,4-difluoro-3-(2-methylpyridin-4-yl)piperidine-1-carboxylate (22-3)

[0382] Under a nitrogen atmosphere, 22-2 (1.5 g, 5.17 mmol) was dissolved in anhydrous dichloromethane (30 ml), and DAST (8.33 g, 51.7 mmol) was slowly added dropwise in an ice-water bath for 4 hours. After the reaction was complete, the reaction was quenched dropwise with saturated sodium bicarbonate aqueous solution, and the mixture was extracted three times with dichloromethane. The organic phase was concentrated and purified by silica gel column chromatography (PE:EA = 2:1) to obtain 22-3 (450 mg).

[0383] 1 H NMR (300 MHz, Chloroform-d) δ 8.48 (d, J = 5.1 Hz, 1H), 7.13 (d, J= 1.8 Hz, 1H), 7.08 (d, J = 5.3 Hz, 1H), 4.23 (s, 2H), 3.41 – 2.91 (m, 3H), 2.58 (s, 3H), 2.17 (dq, J = 10.9, 5.0, 4.0 Hz, 1H), 2.09 – 1.89 (m, 1H), 1.47 (d, J = 1.8 Hz, 9H).

[0384] Step 3: Synthesis of 4-(4,4-difluoropiperidin-3-yl)-2-methylpyridine (22-4)

[0385] Dissolve 22-3 (100 mg, 0.32 mmol) in dichloromethane (5 ml), add 1 ml of dioxane hydrochloride solution, stir for 1 hour, adjust the pH of the solution to neutral after the reaction is complete, extract three times with EA, combine the organic phases, dry with anhydrous sodium sulfate, filter, concentrate the organic phase to obtain crude 22-4 (70 mg), which can be used directly in the next step without purification.

[0386] Step 4: Synthesis of 8-(4-chloro-2-fluorophenyl)-6-(4,4-difluoro-3-(2-methylpyridin-4-yl)piperidin-1-yl)-1,3-dimethylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 22)

[0387] Under N2 protection, DIPEA (93 mg, 0.72 mmol) was added to anhydrous 1,4-dioxane (10 mL) solutions of 22-4 (70 mg, 0.33 mmol) and 1-7 (85 mg, 0.24 mmol), and the reaction was carried out at 100 ºC for 3 h. After the reaction was complete, the mixture was separated by silica gel column chromatography (PE:EA = 1:1) to obtain compound 22 (13 mg).

[0388] 1 H NMR (300 MHz, Chloroform-d) δ 8.47 (d, J = 5.3 Hz, 1H), 7.54 (t, J= 7.9 Hz, 1H), 7.31 (dd, J = 8.5, 1.9 Hz, 1H), 7.24 – 6.94 (m, 3H), 4.96 (t,J = 14.9 Hz, 2H), 3.50 (s, 4H), 3.38 (t, J = 13.0 Hz, 1H), 3.15 (s, 1H), 3.04(s, 3H), 2.60 (s, 3H), 2.27 (s, 2H).

[0389] MS m / z (ES-API+): 531.10 [M+H] +

[0390] Example 23: Synthesis of 8-(2,4-difluorophenyl)-1,3-dimethyl-6-(2-methyl-6-(2-methylpyridin-4-yl)morpholinyl)pyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (compound 23)

[0391]

[0392] Step 1: Synthesis of 2-bromo-1-(2-methylpyridin-4-yl)ethyl-1-one (23-2)

[0393] 23-1 (100 mg, 0.74 mmol) and bromine (18.9 mg, 0.74 mmol) were dissolved in AcOH (containing 33% HBr, 5 mL), and reacted at 40 °C for 1 h, then the temperature was increased to 80 °C and reacted for another 1 h. After the reaction was complete, the mixture was cooled to room temperature, and anhydrous diethyl ether (10 mL) was added. The mixture was stirred for 30 minutes, filtered, and washed with a small amount of anhydrous diethyl ether. The filter cake was collected to obtain crude 23-2 (215 mg), which did not require purification and was directly used for the next step.

[0394] Step 2: Synthesis of 2-(benzyl(2-hydroxypropyl)amino)-1-(2-methylpyridin-4-yl)ethane-1-one (23-3)

[0395] Crude product 23-2 (100 mg, 0.33 mmol) and 1-(benzylamino)prop-2-ol (19.9 mg, 0.73 mmol) were dissolved in THF (8 mL), and DIPEA (225 μL, 1.46 mmol) was added at 0 °C. The mixture was then transferred to room temperature and stirred overnight. After the reaction was complete, the reaction was quenched with water, extracted with ethyl acetate, and the organic layer was dried over anhydrous MgSO4, filtered, and concentrated. Purification by silica gel column chromatography (PE / EA = 600%–100%) yielded 23-3 (140 mg).

[0396] 1 H NMR (300 MHz, Chloroform-d) δ 8.47 (d, J = 5.2 Hz, 1H), 7.40 –7.27 (m, 7H), 4.91 (s, 1H), 4.27 (dtd, J = 14.2, 6.5, 3.9 Hz, 1H), 3.57 (s,2H), 2.84 (dt, J = 11.3, 2.3 Hz, 1H), 2.77 (dd, J = 10.9, 1.8 Hz, 1H), 2.56(s, 3H), 2.17 (d, J = 10.9 Hz, 1H), 1.22 (d, J = 6.3 Hz, 3H).

[0397] Step 3: Synthesis of 4-benzyl-2-methyl-6-(2-methylpyridin-4-yl)-3,4-dihydro-2H-1,4-oxazine (23-4)

[0398] 23-3 (100 mg, 0.73 mmol) was dissolved in anhydrous DCM (5 mL), and TMSOTf (183 μL, 0.99 mmol) was added at 0 °C. The mixture was then transferred to 80 °C and stirred for 16 h. After the reaction was complete, the reaction was quenched with water, extracted with dichloromethane, and the organic layer was dried over anhydrous MgSO4, filtered, and concentrated. 23-4 (59 mg) was purified by silica gel column chromatography (PE / EA = 600%–100%).

[0399] 1 H NMR (300 MHz, Chloroform-d) δ 8.22 (d, J = 5.5 Hz, 1H), 7.37 –7.28 (m, 5H), 7.10 (d, J = 1.6 Hz, 1H), 7.05 – 6.99 (m, 1H), 6.57 (s, 1H),4.19 (t, J = 4.9 Hz, 2H), 4.01 (ddt, J = 9.9, 6.0, 3.0 Hz, 1H), 3.08 (dd, J =12.0, 2.3 Hz, 1H), 2.86 – 2.79 (m, 1H), 2.49 (s, 3H), 1.33 (d, J = 6.4 Hz, 3H).

[0400] Step 4: Synthesis of 2-methyl-6-(2-methylpyridin-4-yl)morpholine (23-5)

[0401] 23-4 (200 mg, 0.71 mmol) was dissolved in methanol (5 mL), and Pd(OH)₂ (120 mg, 0.85 mmol) and ammonium formate (270 mg, 4.26 mmol) were added. The mixture was then stirred at 80 °C for 4 h. After the reaction was complete, the reaction was quenched with saturated sodium bicarbonate aqueous solution, extracted with ethyl acetate, and the organic layer was dried over anhydrous MgSO₄, filtered, and concentrated. 23-5 (69 mg) was purified by silica gel column chromatography (PE / EA = 500%–100%).

[0402] Step 5: Synthesis of 8-(2,4-difluorophenyl)-1,3-dimethyl-6-(2-methyl-6-(2-methylpyridin-4-yl)morpholinyl)pyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (compound 23)

[0403] 16-5 (148 mg, 0.42 mmol), 23-5 (88 mg, 0.45 mmol), XphosPd G4 (32 mg, 0.04 mmol), and t-BuONa (34 mg, 0.35 mmol) were added to a two-necked flask. Anhydrous 1,4-dioxane (8 mL) was added under N2 protection, and the reaction was carried out at 100 °C for 5 hours. After the reaction was completed, the reaction was quenched with water, extracted three times with EA, and the organic layer was dried over anhydrous Na2SO4 and concentrated. The purified compound 23 (20 mg) was obtained by silica gel column chromatography (PE / EA = 800%-300%).

[0404] 1 H NMR (400 MHz, CDCl3) δ 8.55 (d, J = 5.3 Hz, 1H), 7.57 (d, J = 8.4Hz, 1H), 7.42 (s, 1H), 7.39 (s, 1H), 7.34 (d, J = 5.2 Hz, 1H), 7.06 (s, 1H), 6.94 (s, 1H), 4.70 (d, J = 9.9 Hz, 1H), 4.39 (dd, J = 12.4, 2.6 Hz, 1H), 4.18 (d, J = 11.1 Hz, 1H), 3.93 (s, 1H), 3.51 (s, 3H), 3.08 (s, 3H), 2.72 (s, 5H),1.41 (d, J = 6.3 Hz, 3H).

[0405] MS m / z (ES-API+): 494.20 [M+H] +

[0406] Example 24: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-(2-methyl-6-(2-methylpyridin-4-yl)morpholinyl)pyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (compound 24)

[0407]

[0408] 18-2 (460 mg, 1.30 mmol), 23-5 (300 mg, 1.56 mmol), XphosPd G4 (110 mg, 0.13 mmol), and t-BuONa (115 mg, 1.20 mmol) were added to a two-necked flask. Anhydrous 1,4-dioxane (10 mL) was added under N2 protection, and the reaction was carried out at 100 °C for 5 hours. After the reaction was completed, the reaction was quenched with water, extracted three times with EA, and the organic layer was dried over anhydrous Na2SO4 and concentrated under vacuum. The purified compound 24 (15 mg) was obtained by silica gel column chromatography (PE / EA = 600%–300%).

[0409] 1 H NMR (400 MHz, CDCl3) δ 8.53 (d, J = 5.2 Hz, 1H), 7.55 (q, J = 9.0Hz, 1H), 7.42 (s, 1H), 7.35 – 7.29 (m, 2H), 7.23 (d, J = 4.8 Hz, 2H), 4.66(d, J = 10.7 Hz, 1H), 4.34 (d, J = 13.0 Hz, 1H), 4.17 (t, J = 10.8 Hz, 1H), 3.92 (s, 1H), 3.51 (s, 3H), 3.09 (s, 3H), 2.73 (t, J = 11.8 Hz, 2H), 2.64 (s, 3H), 1.40 (d, J = 6.2 Hz, 3H).

[0410] MS m / z (ES-API+): 510.20 [M+H] +

[0411] Example 25: Synthesis of 6-(2-(1-cyclopropyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-4-yl)-8-(2,4-difluorophenyl)-1,3-dimethylpyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (compound 25)

[0412]

[0413] Step 1: Synthesis of 6-(6-(1-cyclopropyl-1H-pyrazol-4-yl)-3,6-dihydro-2H-pyran-4-yl)-8-(2,4-difluorophenyl)-1,3-dimethylpyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (25-1)

[0414] To a mixed solution of 1,4-dioxane (10 mL) and water (2 mL) containing 16-5 (300 mg, 0.89 mmol) and 2-3 (365 mg, 1.15 mmol), Pd(dppf)₂Cl₂·DCM (58 mg, 0.07 mmol) and K₂CO₃ (368 mg, 2.67 mmol) were added. The mixture was then stirred at 80 °C for 5 hours under N₂ protection. After the reaction was complete, the resulting mixture was diluted with water and EA, and extracted three times with EA. The organic layer was dried over anhydrous MgSO₄, filtered, concentrated, and purified by silica gel column chromatography (PE / EA = 600%–300%) to obtain 25-1 (100 mg).

[0415] Step 2: Synthesis of 6-(2-(1-cyclopropyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-4-yl)-8-(2,4-difluorophenyl)-1,3-dimethylpyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (compound 25)

[0416] 25-1 (85 mg, 0.17 mmol) and PtO2 (39 mg, 0.17 mmol) were dissolved in ethanol (6 mL) and reacted at room temperature for 1 hour under a hydrogen atmosphere. After the reaction was complete, the mixture was filtered, and the residue was washed with a small amount of ethanol. The filtrate was concentrated and purified by silica gel column chromatography (PE / EA = 400% - 10%) to obtain 25-1 (40 mg).

[0417] 1H NMR (400 MHz, Chloroform-d) δ 7.96 (s, 1H), 7.66 (q, J = 7.4 Hz,1H), 7.49 (s, 2H), 7.09 (td, J = 9.1, 2.4 Hz, 1H), 7.01–6.89 (m, 1H), 4.52(d, J = 11.0 Hz, 1H), 4.29–4.20 (m, 1H), 3.83–3.71 (m, 1H), 3.58 (s, 1H), 3.53 (s, 3H), 3.30–3.19 (m, 1H), 3.13 (s, 3H), 2.25 (d, J = 12.6 Hz, 1H), 2.03–1.86 (m, 3H), 1.11 (s, 2H), 1.01 (d, J = 6.7 Hz, 2H).

[0418] MS m / z (ES-API+): 494.20 [M+H] +

[0419] Example 26: Synthesis of 8-(4-chloro-2-fluorophenyl)-6-(2-(1-cyclopropyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-4-yl)-1,3-dimethylpyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (compound 26)

[0420]

[0421] Step 1: Synthesis of 8-(4-chloro-2-fluorophenyl)-6-(6-(1-cyclopropyl-1H-pyrazol-4-yl)-3,6-dihydro-2H-pyran-4-yl)-1,3-dimethylpyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (26-1)

[0422] To a mixed solution of 1,4-dioxane (10 mL) and water (2 mL) containing 18-2 (250 mg, 0.71 mmol) and 2-3 (269 mg, 1.70 mmol), Pd(dppf)₂Cl₂·DCM (46 mg, 0.06 mmol) and K₂CO₃ (294 mg, 2.12 mmol) were added. The mixture was then stirred at 80 °C for 5 hours under N₂ protection. After the reaction was complete, the resulting mixture was diluted with water and EA, and extracted three times with EA. The organic layer was dried over anhydrous MgSO₄, filtered, concentrated, and purified by silica gel column chromatography (PE / EA = 600%–300%) to obtain 26-1 (70 mg).

[0423] MS m / z (ES-API+): 508.20 [M+H] + .

[0424] Step 2: Synthesis of 8-(4-chloro-2-fluorophenyl)-6-(2-(1-cyclopropyl-1H-pyrazol-4-yl)tetrahydro-2H-pyran-4-yl)-1,3-dimethylpyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (compound 26)

[0425] Compound 26-1 (65 mg, 0.13 mmol) and PtO2 (29 mg, 0.13 mmol) were dissolved in ethanol (6 mL) and reacted at room temperature for 1 hour under a hydrogen atmosphere. After the reaction was complete, the mixture was filtered, and the residue was washed with a small amount of ethanol. The filtrate was concentrated and purified by silica gel column chromatography (PE / EA = 400% - 10%) to obtain compound 26 (32 mg).

[0426] 1H NMR (400 MHz, Chloroform-d) δ 7.97 (s, 1H), 7.66–7.59 (m, 1H), 7.48 (s, 2H), 7.35 (dd, J = 8.3, 1.9 Hz, 1H), 7.23 (dd, J = 9.5, 2.0 Hz, 1H), 4.52 (d, J = 10.6 Hz, 1H), 4.24 (d, J = 12.5 Hz, 1H), 3.82–3.73 (m, 1H), 3.57 (dt, J = 7.1, 3.5 Hz, 1H), 3.53 (s, 3H), 3.23 (td, J = 11.4, 5.6 Hz, 1H),3.14 (s, 3H), 2.24 (d, J = 13.1 Hz, 1H), 2.02–1.87 (m, 3H), 1.11 (s, 2H), 1.00 (d, J = 7.0 Hz, 2H).

[0427] MS m / z (ES-API+): 510.20 [M+H] +

[0428] Example 27: Synthesis of 1-ethyl-8-(2-fluoro-4-(trifluoromethyl)phenyl)-3-methyl-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholino)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 27)

[0429]

[0430] Step 1: Synthesis of 6-chloro-1-ethyl-8-(2-fluoro-4-(trifluoromethyl)phenyl)-3-methylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (27-1)

[0431] 14-2 (300 mg, 0.80 mmol) was dissolved in anhydrous DMF (7 mL), and K2CO3 (332 mg, 2.4 mmol) was added with stirring. Iodoethane (250 mg, 1.60 mmol) was then added dropwise, and the reaction was carried out at 80 °C for 4 hours. After the reaction was complete, the reaction was quenched with water, and the mixture was extracted three times with ethyl acetate. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. Purification by silica gel column chromatography (PE / EA = 800%–400%) yielded 27-1 (73 mg).

[0432] 1H NMR (300 MHz, CDCl3) δ 7.91 (t, J = 7.5 Hz, 1H), 7.69 (d, J = 8.0Hz, 1H), 7.57 (d, J = 9.7 Hz, 1H), 4.51 (q, J = 7.1 Hz, 2H), 3.68 (d, J = 2.3Hz, 3H), 1.51 (s, 3H).

[0433] Step 2: Synthesis of 1-ethyl-8-(2-fluoro-4-(trifluoromethyl)phenyl)-3-methyl-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholino)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 27)

[0434] Compound 27-1 (73 mg, 0.18 mmol) was dissolved in DMSO (5 mL), and DIPEA (117 mg, 0.91 mmol) and 1-8 (39 mg, 0.24 mmol) were added. The mixture was reacted at 100 °C for 2 hours. After the reaction was complete, the reaction was quenched with water, and the mixture was extracted three times with ethyl acetate. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. The solution was purified by silica gel column chromatography (PE / EA = 800%–400%) to give compound 27 (51 mg).

[0435] 1H NMR (300 MHz, CDCl3) δ 7.93 (t, J = 7.4 Hz, 1H), 7.72 (d, J = 10.5Hz, 2H), 7.61 (d, J = 6.7 Hz, 2H), 4.98 (d, J = 13.4 Hz, 1H), 4.83 (d, J =13.5 Hz, 1H), 4.75 (dd, J = 10.5, 2.8 Hz, 1H), 4.48 (q, J = 7.1 Hz, 2H), 4.29– 4.18 (m, 1H), 4.09 (s, 3H), 4.03–3.89 (m, 1H), 3.70 (s, 3H), 3.47 (t, J =12.0 Hz, 1H), 3.38 (dd, J = 13.4, 10.2 Hz, 1H), 1.54 (s, 3H).

[0436] MS m / z (ES-API+): 534.20 [M+H] +

[0437] Example 28: Synthesis of 8-(2-fluoro-4-(trifluoromethyl)phenyl)-3-methyl-1-(methyl-D3)-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholino)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 28)

[0438]

[0439] Step 1: Synthesis of 6-chloro-8-(2-fluoro-4-(trifluoromethyl)phenyl)-3-methyl-1-(methyl-d3)pyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (28-1)

[0440] 14-2 (2.5 g, 6.68 mmol) was dissolved in anhydrous DMF solution (15 mL), and NaH (240 mg, 10.0 mmol) was added at 0 °C. The mixture was then heated to room temperature and reacted for 20 min. CD3I (1.06 g, 7.35 mmol) was then added, and the mixture was reacted at room temperature for 3 h. After the reaction was complete, the reaction was quenched with water, and the mixture was extracted three times with ethyl acetate. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. The solution was purified by silica gel column chromatography (PE / EA = 800%–400%) to obtain 28-1 (1.2 g).

[0441] Step 2: Synthesis of 8-(2-fluoro-4-(trifluoromethyl)phenyl)-3-methyl-1-(methyl-D3)-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholino)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 28)

[0442] Compound 28-1 (101.8 mg, 0.26 mmol) was dissolved in DMSO (3 mL), and DIPEA (103 mg, 0.76 mmol) and 1-8 (45 mg, 0.26 mmol) were added. The mixture was reacted at 80 °C for 3 hours. After the reaction was complete, the reaction was quenched with water, and the mixture was extracted three times with ethyl acetate. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. The solution was purified by silica gel column chromatography (PE / EA = 800%–400%) to give compound 28 (50 mg).

[0443] 1H NMR (400 MHz, Chloroform-d) δ 7.77 (t, J = 7.4 Hz, 1H), 7.63 –7.58 (m, 1H), 7.52 (s, 1H), 7.47 (dd, J = 9.5, 1.6 Hz, 1H), 7.42 (s, 1H),4.71 (d, J = 13.5 Hz, 1H), 4.56 (t, J = 11.8 Hz, 2H), 4.09 – 4.03 (m, 1H), 3.89 (s, 3H), 3.76 (t, J = 11.4 Hz, 1H), 3.51 (s, 3H), 3.35 – 3.13 (m, 2H).

[0444] MS m / z (ES-API+): 523.20 [M+H] +

[0445] Example 29: Synthesis of 8-(2-fluoro-4-(trifluoromethyl)phenyl)-1-methyl-3-(methyl-D3)-6-((2R)-2-methyl-6-(2-methylpyridin-4-yl)morpholino)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 29)

[0446]

[0447] Step 1: Synthesis of (S)-2-(benzyl(2-hydroxypropyl)amino)-1-(2-methylpyridin-4-yl)ethyl-1-one (29-1)

[0448] Crude product 23-2 (100 mg, 0.33 mmol) and (S)-1-(benzylamino)prop-2-ol (19.9 mg, 0.73 mmol) were dissolved in THF (8 mL), and DIPEA (225 μL, 1.46 mmol) was added at 0 °C. The mixture was then transferred to room temperature and stirred overnight. After the reaction was complete, the reaction was quenched with water, extracted with ethyl acetate, and the organic layer was dried over anhydrous MgSO4, filtered, and concentrated. Purification by silica gel column chromatography (PE / EA = 600%–100%) yielded 29-1 (140 mg).

[0449] 1H NMR (300 MHz, Chloroform-d) δ 8.47 (d, J = 5.2 Hz, 1H), 7.40 –7.27 (m, 7H), 4.91 (s, 1H), 4.27 (dtd, J = 14.2, 6.5, 3.9 Hz, 1H), 3.57 (s,2H), 2.84 (dt, J = 11.3, 2.3 Hz, 1H), 2.77 (dd, J = 10.9, 1.8 Hz, 1H), 2.56(s, 3H), 2.17 (d, J = 10.9 Hz, 1H), 1.22 (d, J = 6.3 Hz, 3H).

[0450] Step 2: Synthesis of (R)-4-benzyl-2-methyl-6-(2-methylpyridin-4-yl)-3,4-dihydro-2H-1,4-oxazine (29-2)

[0451] 29-1 (100 mg, 0.73 mmol) was dissolved in anhydrous DCM (5 mL), and TMSOTf (183 μL, 0.99 mmol) was added at 0 °C. The mixture was then transferred to 80 °C and stirred for 16 h. After the reaction was complete, the reaction was quenched with water, extracted with dichloromethane, and the organic layer was dried over anhydrous MgSO4, filtered, and concentrated. 29-2 (59 mg) was purified by silica gel column chromatography (PE / EA = 600%–100%).

[0452] 1 H NMR (300 MHz, Chloroform-d) δ 8.22 (d, J = 5.5 Hz, 1H), 7.37–7.28(m, 5H), 7.10 (d, J = 1.6 Hz, 1H), 7.05 – 6.99 (m, 1H), 6.57 (s, 1H), 4.19(t, J = 4.9 Hz, 2H), 4.01 (ddt, J = 9.9, 6.0, 3.0 Hz, 1H), 3.08 (dd, J =12.0, 2.3 Hz, 1H), 2.86 – 2.79 (m, 1H), 2.49 (s, 3H), 1.33 (d, J = 6.4 Hz, 3H).

[0453] Step 3: Synthesis of (2R)-2-methyl-6-(2-methylpyridin-4-yl)morpholine (29-3)

[0454] Compound 29-2 (200 mg, 0.71 mmol) was dissolved in methanol (5 mL), and Pd(OH)₂ (120 mg) and ammonium formate (270 mg, 4.26 mmol) were added. The mixture was then stirred at 80 °C for 4 h. After the reaction was complete, the reaction was quenched with saturated sodium bicarbonate aqueous solution, and the mixture was extracted three times with ethyl acetate. The organic layer was dried over anhydrous MgSO₄, filtered, and concentrated. Compound 29-3 (69 mg) was purified by silica gel column chromatography (PE / EA = 500%–100%).

[0455] Step 4: Synthesis of 8-(2-fluoro-4-(trifluoromethyl)phenyl)-1-methyl-3-(methyl-D3)-6-((2R)-2-methyl-6-(2-methylpyridin-4-yl)morpholino)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 29)

[0456] 13-3 (47 mg, 0.12 mmol) was dissolved in DMSO (10 mL), and DIPEA (0.5 mL) and 29-3 (30 mg, 0.16 mmol) were added. The mixture was reacted at 80 °C for 3 hours. After the reaction was complete, the reaction was quenched with water, and the mixture was extracted three times with ethyl acetate. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. The solution was purified by silica gel column chromatography (PE / EA = 800%–400%) to give compound 29 (22 mg).

[0457] 1 H NMR (300 MHz, Chloroform-d) δ 8.50 (d, J = 5.2 Hz, 1H), 7.77 (t, J= 7.4 Hz, 1H), 7.62 (d, J = 8.1 Hz, 1H), 7.48 (d, J = 9.5 Hz, 1H), 7.28 (s,1H), 7.20 (d, J = 5.3 Hz, 1H), 4.76 (dd, J = 26.4, 13.3 Hz, 2H), 4.57 (d, J =10.5 Hz, 1H), 3.83 (s, 1H), 3.03 (s, 3H), 2.83 (td, J = 13.9, 10.6 Hz, 2H),2.60 (s, 3H), 1.37 (d, J = 6.1 Hz, 3H).

[0458] MS m / z (ES-API+): 548.20 [M+H] +

[0459] Example 30: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-(2-(2-methylpyridin-4-yl)morpholino)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 30)

[0460]

[0461] Step 1: Synthesis of 4-(tert-butyl)-2-(1,3-dioxoisoindoline-2-yl)morpholine-2,4-dicarboxylic acid ester (30-2)

[0462] 30-1 (24.3 g, 148.9 mmol) and 4-(tert-butyloxycarbonyl)morpholine-2-carboxylic acid (35 g, 148.9 mmol) were added to DCM (200 mL), followed by DMAP (1.85 g, 14.9 mmol) and DIC (20.1 g, 159.5 mmol). The reaction mixture was stirred at room temperature for 3 h. Subsequently, 1000 mL of water was added to quench the reaction, followed by extraction with DCM (200 mL * 3), washing with saturated brine, drying over anhydrous sodium sulfate, filtering, concentrating the filtrate under reduced pressure, and purifying by column chromatography (ethyl acetate / petroleum ether = 1 / 4) to obtain 30-2 (15 g).

[0463] Step 2: Synthesis of [(bipy)2Ni2(μ-Cl)2Cl2(H2O)2] (30-4)

[0464] 30-3 (5 g, 32.01 mmol) and NiCl2·6H2O (7.59 g, 32.01 mmol) were added to EtOH (50 mL). The reaction mixture was stirred at room temperature for 18 h. The reaction mixture was then filtered, washed with ethanol (40 mL * 3), and the filter cake was dried to obtain 30-4 (4 g).

[0465] Step 3: Synthesis of tert-butyl 2-(2-methylpyridin-4-yl)morpholine-4-carboxylic acid (30-5)

[0466] 4-Iodo-2-methylpyridine (10 g, 45.66 mmol), 30-2 (25.78 g, 68.49 mmol), 30-4 (1.73 g, 4.57 mmol), and Zn powder (23.38 g, 365.26 mmol) were added to a 500 mL three-necked flask. The mixture was purged with nitrogen three times, and DMA (100 mL) was added at -5 °C, followed by the dropwise addition of TMSCl (14.79 g, 136.97 mmol). The reaction mixture was stirred in an ice-water bath for 2 h. After the reaction was complete, the mixture was quenched with deionized water (20 mL), diluted with H2O (60 mL), extracted three times with organic solvents DCM and MeOH (DCM:MeOH = 10:1, 200 mL), washed with saturated brine (300 mL * 3), and dried over anhydrous sodium sulfate. The filtrate was filtered, concentrated under reduced pressure, and purified by silica gel column chromatography (ethyl acetate / petroleum ether = 1 / 5) to obtain 30-5 (7 g).

[0467] Step 4: Synthesis of 2-(2-methylpyridin-4-yl)morpholine (30-6)

[0468] Trifluoroacetic acid (1 mL) was added to a solution of 30-5 (700 mg, 2.51 mmol) in dichloromethane (4 mL), and the mixture was reacted at room temperature for 2 h. After the reaction was complete, the mixture was diluted with sodium bicarbonate aqueous solution (5 mL), extracted with dichloromethane (5 mL x 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated, and purified by silica gel column chromatography (PE:EA = 1:1) to obtain 30-6 (400 mg).

[0469] Step 5: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-(2-(2-methylpyridin-4-yl)morpholino)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 30)

[0470] Dissolve compound 1-7 (71 mg, 0.2 mmol) in 6 mL of DMSO solution. Add DIPEA (0.3 mL) and 30-6 (43 mg, 0.24 mmol), and react at 80 °C for 3 hours. Quench the reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, elute with 800%–400% PE / EA solution, and separate to obtain compound 30 (50 mg).

[0471] 1H NMR (300 MHz, CDCl3) δ 8.51 (s, 1H), 7.56 (d, J = 9.1 Hz, 1H), 7.35 (s, 4H), 4.74 (dd, J = 41.8, 13.4 Hz, 2H), 4.55 (s, 1H), 3.80 (s, 2H), 3.52(s, 3H), 3.26 (s, 2H), 3.05 (s, 3H), 2.62 (s, 3H).

[0472] MS m / z (ES-API+): 497.10 [M+H] +

[0473] Example 31: Synthesis of 8-(2-fluoro-4-(trifluoromethyl)phenyl)-1,3-dimethyl-6-((2R)-2-methyl-6-(2-methylpyridin-4-yl)morpholinyl)pyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (compound 31)

[0474]

[0475] 20-2 (168 mg, 0.50 mmol), 29-3 (100 mg, 0.60 mmol), XphosPd G4 (37 mg, 0.05 mmol), and t-BuONa (38 mg, 0.45 mmol) were added to a two-necked flask. Anhydrous 1,4-dioxane (8 mL) was added under N2 protection, and the reaction was carried out at 100 °C for 9 hours. The reaction was quenched with water, extracted three times with EtOAc, and the combined organic layers were dried over anhydrous Na2SO4 and concentrated under vacuum. The mixture was purified on a silica gel column and eluted with 800%–300% PE / EA solution to give compound 31 (20 mg).

[0476] 1H NMR (300 MHz, CDCl3) δ 8.43 (d, J = 5.1 Hz, 1H), 7.67 (d, J = 7.7Hz, 1H), 7.50 (s, 1H), 7.38 (s, 1H), 7.24 – 6.96 (m, 3H), 4.55 (d, J = 10.5Hz, 1H), 4.23 (d, J = 12.9 Hz, 1H), 4.08 (s, 1H), 3.82 (s, 1H), 3.42 (s, 3H), 2.98 (s, 3H), 2.66 (q, J = 9.5 Hz, 2H), 2.51 (s, 3H), 1.31 (d, J = 6.1 Hz,3H).

[0477] MS m / z (ES-API+): 544.10 [M+H] +

[0478] Example 32: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-((2R)-2-methyl-6-(2-methylpyridin-4-yl)morpholinyl)pyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (compound 32)

[0479]

[0480] 18-2 (153 mg, 0.43 mmol), 29-3 (100 mg, 0.52 mmol), XphosPd G4 (37 mg, 0.05 mmol), and t-BuONa (38 mg, 0.45 mmol) were added to a two-necked flask. Anhydrous 1,4-dioxane (8 mL) was added under N2 protection, and the reaction was carried out at 100 °C for 9 hours. The reaction was quenched with water, extracted three times with EtOAc, and the combined organic layers were dried over anhydrous Na2SO4 and concentrated under vacuum. The mixture was purified on a silica gel column and eluted with 800%–300% PE / EA solution to give compound 32 (23 mg).

[0481] 1H NMR (400 MHz, CDCl3) δ 8.53 (d, J = 5.2 Hz, 1H), 7.55 (q, J = 9.0Hz, 1H), 7.42 (s, 1H), 7.35 – 7.29 (m, 2H), 7.23 (d, J = 4.8 Hz, 2H), 4.66(d, J = 10.7 Hz, 1H), 4.34 (d, J = 13.0 Hz, 1H), 4.17 (t, J = 10.8 Hz, 1H), 3.92 (s, 1H), 3.51 (s, 3H), 3.09 (s, 3H), 2.73 (t, J = 11.8 Hz, 2H), 2.64 (s, 3H), 1.40 (d, J = 6.2 Hz, 3H).

[0482] MS m / z (ES-API+): 510.20 [M+H] +

[0483] Example 33: Synthesis of 8-(2,4-difluorophenyl)-1,3-dimethyl-6-((2R)-2-methyl-6-(1-methyl-1H-pyrazol-4-yl)morpholinyl)pyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (compound 33)

[0484]

[0485] 16-5 (155 mg, 0.46 mmol), 3-7 (100 mg, 0.55 mmol), XphosPd G4 (40 mg, 0.05 mmol), and t-BuONa (40 mg, 0.42 mmol) were added to a two-necked flask. Anhydrous 1,4-dioxane (8 mL) was added under N2 protection, and the reaction was carried out at 100 °C for 9 hours. The reaction was quenched with water, extracted three times with EtOAc, and the combined organic layers were dried over anhydrous Na2SO4 and concentrated under vacuum. The mixture was purified on a silica gel column and eluted with 800%–300% PE / EA solution to give compound 33 (70 mg).

[0486] 1H NMR (300 MHz, CDCl3) δ 7.60 (d, J = 10.4 Hz, 2H), 7.48 (s, 1H), 7.39 (s, 1H), 7.05 (t, J = 8.6 Hz, 1H), 6.92 (td, J = 9.5, 2.4 Hz, 1H), 4.67(d, J = 9.7 Hz, 1H), 4.32 (t, J = 12.8 Hz, 1H), 4.13 (t, J = 12.8 Hz, 1H), 3.94 (s, 3H), 3.88 (s, 1H), 3.51 (s, 3H), 3.08 (s, 3H), 2.90 (t, J = 11.7 Hz,1H), 2.70 (dd, J = 12.6, 10.5 Hz, 1H), 1.34 (d, J = 6.2 Hz, 3H).

[0487] MS m / z (ES-API+): 483.10 [M+H] +

[0488] Example 34: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-((2R)-2-methyl-6-(1-methyl-1H-pyrazol-4-yl)morpholinyl)pyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione (compound 34)

[0489]

[0490] Step 1: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-((2R)-2-methyl-6-(1-methyl-1H-pyrazol-4-yl)morpholinyl)pyrido[3,4-d]pyrimidine-2,4(1H,3H)-dione

[0491] 18-2 (81 mg, 0.23 mmol), 3-7 (50 mg, 0.28 mmol), Xphos PdG4 (20 mg, 0.02 mmol), and t-BuONa (20 mg, 0.21 mmol) were added to a two-necked flask. Anhydrous 1,4-dioxane (8 mL) was added under N2 protection, and the reaction was carried out at 100 °C for 9 hours. The reaction was quenched with water, extracted three times with EtOAc, and the combined organic layers were dried over anhydrous Na2SO4 and concentrated under vacuum. The mixture was purified on a silica gel column and eluted with 800%–300% PE / EA solution to give compound 34 (20 mg).

[0492] 1H NMR (400 MHz, CDCl3) δ 7.53 (s, 1H), 7.44 (s, 1H), 7.37 (s, 1H), 7.29 (d, J = 2.0 Hz, 1H), 7.26 (s, 1H), 7.18 (dd, J = 9.5, 2.0 Hz, 1H), 4.64(d, J = 10.6 Hz, 1H), 4.35 – 4.22 (m, 1H), 4.11 (t, J = 16.0 Hz, 1H), 3.89(s, 3H), 3.86 (s, 1H), 3.49 (s, 3H), 3.06 (s, 3H), 2.88 (t, J = 12.0 Hz, 1H),2.67 (dd, J = 12.6, 10.5 Hz, 1H), 1.32 (d, J = 6.2 Hz, 3H).

[0493] MS m / z (ES-API+): 499.10 [M+H] +

[0494] Example 35: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-(5-(2-methylpyridin-4-yl)-4-oxa-7-azaspiro[2.5]octane-7-yl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 35)

[0495]

[0496] Step 1: Synthesis of 2-(benzyl((1-hydroxycyclopropyl)methyl)amino)-1-(2-methylpyridin-4-yl)ethane-1-one (35-1)

[0497] Crude product 23-2 (2.4 g, 8.76 mmol) and 1-((benzylamino)methyl)cyclopropane-1-ol (2.3 g, 13.14 mmol) were dissolved in THF (25 mL). DIPEA (3.1 mL, 26.28 mmol) was added at 0 °C. After the addition was complete, the mixture was transferred to room temperature and stirred overnight. After the reaction was complete, the reaction was quenched with water, and the mixture was extracted three times with ethyl acetate. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. Purification was performed on a silica gel column, eluting with 600%–100% PE / EA solution to obtain 35-1 (1.24 g).

[0498] Step 2: Synthesis of 7-benzyl-5-(2-methylpyridin-4-yl)-4-oxa-7-azaspiro[2.5]oct-5-ene (35-2)

[0499] 35-1 (1.24 g, 3.99 mmol) was dissolved in anhydrous DCM (20 mL), and TMSOTf (2.17 mL, 11.99 mmol) was added at 0 °C. After the addition was complete, the mixture was transferred to 80 °C and stirred for 16 h. After the reaction was complete, the reaction was quenched with water, and the mixture was extracted three times with dichloromethane. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. The solution was purified on a silica gel column and eluted with 600%–100% PE / EA solution to obtain 35-2 (220 mg).

[0500] Step 3: Synthesis of 5-(2-methylpyridin-4-yl)-4-oxa-7-azaspiro[2.5]octane (35-3)

[0501] 35-2 (100 mg, 0.35 mmol) was dissolved in methanol (3 mL), and 5% palladium on carbon (40 mg), 10% palladium hydroxide on carbon (50 mg), and ammonium formate (200 mg, 3.17 mmol) were added at room temperature. The mixture was then transferred to 80 °C and stirred for 4 h. After the reaction was complete, the reaction was quenched with saturated sodium bicarbonate aqueous solution, and the mixture was extracted three times with ethyl acetate. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. The solution was purified on a silica gel column and eluted with 500%–100% PE / EA solution to obtain 35-3 (22.4 mg).

[0502] Step 4: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-(5-(2-methylpyridin-4-yl)-4-oxa-7-azaspiro[2.5]octane-7-yl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 35)

[0503] Compound 1-7 (40.0 mg, 0.11 mmol) was dissolved in DMSO (3 mL), and DIPEA (42.6 mg, 0.33 mmol) and 5-(2-methylpyridin-4-yl)-4-oxa-7-azaspiro[2.5]octane (22.4 mg, 0.11 mmol) were added. The mixture was reacted at 100 °C for 2 hours. The reaction was quenched with water, and the mixture was extracted three times with ethyl acetate. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. The mixture was purified on a silica gel column and eluted with 800%–400% PE / EA solution to obtain compound 35 (21 mg).

[0504] 1H NMR (300 MHz, CDCl3) δ 8.51 (d, J = 5.1 Hz, 1H), 7.57 (t, J = 8.1Hz, 1H), 7.36 (dd, J = 8.3, 2.0 Hz, 1H), 7.25 (d, J = 1.8 Hz, 2H), 7.17 (d, J= 5.2 Hz, 1H), 4.91 (d, J = 13.5 Hz, 1H), 4.71 (d, J = 9.1 Hz, 1H), 4.22 –4.10 (m, 1H), 3.78 – 3.69 (m, 1H), 3.54 (s, 3H), 3.07 (s, 4H), 2.60 (s, 3H),1.20 – 1.02 (m, 2H), 0.99 – 0.86 (m, 2H).

[0505] MS m / z (ES-API+): 523.10 [M+H] +

[0506] Example 36: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-((2R)-2-methyl-6-(2-methylpyridin-4-yl)morpholino)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 36)

[0507]

[0508] Step 1: Synthesis of (R)-2-(benzyl(2-hydroxypropyl)amino)-1-(2-methylpyridin-4-yl)ethyl-1-one (36-1)

[0509] Crude product 23-2 (100 mg, 0.33 mmol) and (R)-1-(benzylamino)prop-2-ol (19.9 mg, 0.73 mmol) were dissolved in THF (8 mL). DIPEA (225 μL, 1.46 mmol) was added at 0 °C. After the addition was complete, the mixture was transferred to room temperature and stirred overnight. After the reaction was complete, the reaction was quenched with water, and the mixture was extracted three times with ethyl acetate. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. Purification was performed on a silica gel column, eluting with 600%–100% PE / EA solution to obtain 36-1 (140 mg).

[0510] Step 2: Synthesis of (R)-4-benzyl-2-methyl-6-(2-methylpyridin-4-yl)-3,4-dihydro-2H-1,4-oxazine (36-2)

[0511] 36-1 (100 mg, 0.73 mmol) was dissolved in anhydrous DCM (5 mL), and TMSOTf (183 μL, 0.99 mmol) was added at 0 °C. After the addition was complete, the mixture was transferred to 80 °C and stirred for 16 h. After the reaction was complete, the reaction was quenched with water, and the mixture was extracted three times with dichloromethane. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. The solution was purified on a silica gel column and eluted with 600%–100% PE / EA solution to obtain 36-2 (59 mg).

[0512] Step 3: Synthesis of (R)-2-methyl-6-(2-methylpyridin-4-yl)morpholine (36-3)

[0513] 36-2 (200 mg, 0.71 mmol) was dissolved in methanol (5 mL), and 10% palladium hydroxide on carbon (120 mg) and ammonium formate (270 mg, 4.26 mmol) were added at room temperature. The mixture was then stirred at 80 °C for 4 h. After the reaction was complete, the reaction was quenched with saturated sodium bicarbonate aqueous solution, and the mixture was extracted three times with ethyl acetate. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. The solution was purified on a silica gel column and eluted with 500%–100% PE / EA solution to obtain 36-3 (69 mg).

[0514] Step 4: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-((2R)-2-methyl-6-(2-methylpyridin-4-yl)morpholinyl)pyrimidino-5,4−dpyrimidin-2,4(1H,3H)-dione (36-4)

[0515] Dissolve compound 1-7 (158 mg, 0.44 mmol) in 5 mL of DMSO solution. Add DIPEA (173 mg, 1.3 mmol) and 36-3 (103 mg, 0.54 mmol), and react at 100 °C for 2 h. Quench the reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 800%–400% PE / EA solution to obtain compound 36 (158 mg).

[0516] 1H NMR (300 MHz, Chloroform-d) δ 8.49 (d, J = 5.2 Hz, 1H), 7.58 (t, J= 8.0 Hz, 1H), 7.41 –7.28 (m, 2H), 7.26 – 7.15 (m, 2H), 4.76 (dd, J = 25.7,13.4 Hz, 2H), 4.60 – 4.49 (m, 1H), 3.88 – 3.77 (m, 1H), 3.51 (s, 3H), 3.05(s, 3H), 2.90 – 2.73 (m, 2H), 2.59 (s, 3H), 1.37 (d, J = 6.2 Hz, 3H).

[0517] MS m / z (ES-API+): 511.10 [M+H] +

[0518] Example 37: Synthesis of 8-(2,4-difluorophenyl)-1,3-dimethyl-6-(6-(1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-3,6-dihydro-2H-pyran-4-yl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 37)

[0519]

[0520] Add 37-1 (190 mg, 0.60 mmol), 8-6 (140 mg, 0.40 mmol), Pd(dppf)Cl2 (30 mg, 0.04 mmol), and potassium carbonate (166 mg, 1.20 mmol) to a 50 mL two-necked flask. Under nitrogen atmosphere, add 10 mL of 1,4-dioxane and 2 mL of water to dissolve. o The reaction was heated at C for 4 hours. After the reaction was complete, the reaction mixture was diluted with aqueous solution, extracted three times with ethyl acetate, and the organic layer was dried over anhydrous MgSO4, filtered, and concentrated. The mixture was purified on a silica gel column and eluted with 500%–50% PE / EA solution to give compound 37 (80 mg).

[0521] 1H NMR (300 MHz, CDCl3) δ 7.79 (s, 1H), 7.44 (d, J = 18.9 Hz, 2H), 7.22 – 6.92 (m, 2H), 6.76 (s, 1H), 5.22 (s, 1H), 4.18 (s, 1H), 3.92 (s, 1H), 3.71 – 3.48 (m, 6H), 3.17 (s, 3H), 2.90 (s, 2H), 1.30 (s, 1H).

[0522] MS m / z (ES-API+): 494.10 [M+H]+

[0523] Example 38: Synthesis of 1-ethyl-8-(2-fluoro-4-(trifluoromethyl)phenyl)-3-methyl-6-((2R)-2-methyl-6-(1-methyl-1H-pyrazol-4-yl)morpholinyl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 38)

[0524]

[0525] Dissolve 27-1 (100 mg, 0.25 mmol) in DMSO (4 mL). Add DIPEA (0.2 mL) and 3-7 (67 mg, 0.37 mmol), and react at 100 °C for 5 h. Quench the reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 200%–40% PE / EA solution to obtain compound 38 (90 mg).

[0526] 1H NMR (400 MHz, CDCl3) δ 7.79 (t, J = 7.3 Hz, 1H), 7.64 (s, 1H), 7.57(d, J = 8.0 Hz, 1H), 7.52 (s, 1H), 7.50 – 7.44 (m, 1H), 4.93 (d, J = 13.2 Hz,1H), 4.81 (d, J = 13.2 Hz, 1H), 4.71 – 4.61 (m, 1H), 4.34 (p, J = 6.9 Hz,2H), 3.99 (d, J = 2.8 Hz, 3H), 3.85 (d, J = 7.5 Hz, 1H), 3.55 (d, J = 2.4 Hz,3H), 3.08 – 2.97 (m, 1H), 2.81 (dd, J = 13.4, 10.5 Hz, 1H), 1.40 (td, J =7.1, 2.5 Hz, 3H), 1.34 (d, J = 6.2 Hz, 3H).

[0527] MS m / z (ES-API+): 548.10 [M+H]+

[0528] Example 39: Synthesis of 8-(4-chloro-2-fluorophenyl)-1-ethyl-3-methyl-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholinyl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 39)

[0529]

[0530] Step 1: Synthesis of 6-chloro-8-(4-chloro-2-fluorophenyl)-1-ethyl-3-methylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (39-1)

[0531] 27-1 (300 mg, 0.88 mmol) was dissolved in anhydrous DMF (7 mL), and K2CO3 (366 mg, 2.65 mmol) was added with stirring. Then, iodoethane (274 mg, 1.76 mmol) was added dropwise, and the reaction was carried out at 80 °C for 4 hours. After the reaction was complete, the reaction was quenched with water, and the mixture was extracted three times with ethyl acetate. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. Purification by silica gel column chromatography (PE / EA = 800%–400%) yielded 39-1 (86 mg).

[0532] Step 2: Synthesis of 8-(4-chloro-2-fluorophenyl)-1-ethyl-3-methyl-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholinyl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 39)

[0533] Dissolve 39-1 (100 mg, 0.27 mmol) in DMSO (5 mL). Add DIPEA (0.2 mL) and 1-8 (55 mg, 0.405 mmol), and react at 100 °C for 5 hours. Quench the reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 200%–50% PE / EA solution to obtain compound 39 (85 mg).

[0534] 1 H NMR (300 MHz, CDCl3) δ 7.61 (d, J = 8.3 Hz, 2H), 7.51 (s, 1H), 7.21 (d, J = 9.8 Hz, 2H), 4.82 (d, J = 13.2 Hz, 1H), 4.62 (t, J = 12.9 Hz, 2H), 4.32 (q, J = 7.1 Hz, 2H), 4.07 (d, J = 11.6 Hz, 1H), 3.97 (s, 3H), 3.78 (t, J= 11.4 Hz, 1H), 3.53 (s, 3H), 3.26 (dt, J = 24.5, 11.5 Hz, 2H), 1.37 (t, J =7.1 Hz, 3H).

[0535] MS m / z (ES-API+): 500.10 [M+H]+

[0536] Example 40: Synthesis of 8-(4-chloro-2-fluorophenyl)-1-ethyl-3-methyl-6-((2R)-2-methyl-6-(1-methyl-1H-pyrazol-4-yl)morpholinyl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 40)

[0537]

[0538] Dissolve 39-1 (100 mg, 0.27 mmol) in DMSO (5 mL). Add DIPEA (0.2 mL) and 3-7 (73 mg, 0.405 mmol), and react at 100 °C for 5 h. Quench the reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 200%–50% PE / EA solution to obtain compound 40 (81 mg).

[0539] 1 H NMR (400 MHz, CDCl3) δ 7.63 – 7.54 (m, 2H), 7.46 (s, 1H), 7.28 (d,J = 2.0 Hz, 1H), 7.21 (dd, J = 9.7, 2.0 Hz, 1H), 4.89 (d, J = 13.2 Hz, 1H), 4.79 (d, J = 13.1 Hz, 1H), 4.60 (dd, J = 10.9, 2.7 Hz, 1H), 4.32 (q, J = 7.1Hz, 2H), 3.91 (d, J = 1.9 Hz, 3H), 3.81 (ddd, J = 10.6, 6.3, 2.6 Hz, 1H),3.52 (s, 3H), 3.01 (dd, J = 13.2, 10.9 Hz, 1H), 2.84 – 2.70 (m, 1H), 1.37 (t,J = 7.1 Hz, 3H), 1.31 (d, J = 6.2 Hz, 3H).

[0540] MS m / z (ES-API+): 514.10 [M+H]+

[0541] Example 41: Synthesis of (S)-8-(2,4-difluorophenyl)-1-ethyl-3-methyl-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholinyl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 41-P1 or compound 41-P2) and (R)-8-(2,4-difluorophenyl)-1-ethyl-3-methyl-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholinyl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 41-P1 or compound 41-P2)

[0542]

[0543]

[0544] Step 1: Synthesis of (S)-2-(1-methyl-1H-pyrazol-4-yl)morpholine (1-8-P1 or 1-8-P2) and (R)-2-(1-methyl-1H-pyrazol-4-yl)morpholine (1-8-P1 or 1-8-P2)

[0545] Commercially available 1-8 (5.0 g) was purified and separated by supercritical fluid chromatography (SCLC) (Column: ChiralPak AD, column length 250 mm, inner diameter 50 mm, particle size 10 μm; Mobile phase A: supercritical carbon dioxide, Mobile phase B: methanol (containing 0.1% ammonia); Gradient: Mobile phase B maintained at 35%; Flow rate: 150 mL / min; Column temperature: 25℃; Back pressure: 100 bar) to obtain 1-8-P1 (2.27 g) and 1-8-P2 (2.25 g). The two title products were then further analyzed using the following chiral SFC method.

[0546] Chiral SFC Analysis Method:

[0547] Chromatographic column: ChiralPak AD, column length 100 mm, inner diameter 4.6 mm, particle size 3.0 μm; mobile phase A: supercritical carbon dioxide, mobile phase B: methanol (containing 0.1% ammonia); gradient: mobile phase B from 5% to 40%; flow rate: 2.0 mL / min; column temperature: 40℃; back pressure: 100 bar; detection wavelength: 210 nm.

[0548] 1-8-P1 (enantiomer peak 1): Chiral SFC elution time was 2.648 min; MS m / z (ES-API+): 168.04 [M+H] + ;

[0549] 1-8-P2 (enantiomer peak 2): Chiral SFC elution time was 3.035 min; MS m / z (ES-API+): 168.04 [M+H] + .

[0550] Step 2: Synthesis of 6-chloro-8-(2,4-difluorophenyl)-1-ethyl-3-methylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (41-1)

[0551] 8-5 (300 mg, 0.92 mmol) was dissolved in a mixture of anhydrous DMF (5 mL) and acetonitrile (5 mL) (7 mL), and K2CO3 (383 mg, 2.78 mmol) was added with stirring. Iodoethane (287 mg, 1.84 mmol) was then added dropwise, and the reaction was carried out at 80 °C for 4 hours. After the reaction was complete, the reaction was quenched with water, and the mixture was extracted three times with ethyl acetate. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. Purification by silica gel column chromatography (PE / EA = 800%–400%) yielded 41-1 (80 mg).

[0552] Step 3: Synthesis of (S)-8-(2,4-difluorophenyl)-1-ethyl-3-methyl-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholinyl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 41-P1 or compound 41-P2) and (R)-8-(2,4-difluorophenyl)-1-ethyl-3-methyl-6-(2-(1-methyl-1H-pyrazol-4-yl)morpholinyl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 41-P1 or compound 41-P2).

[0553] Dissolve 41-1 (100 mg, 0.28 mmol) in DMSO (5 mL). Add DIPEA (0.2 mL) and 1-8-P1 (72 mg, 0.42 mmol), and react at 100 °C for 5 hours. Quench the reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 200%–50% PE / EA solution to obtain compound 41-P1 (80 mg).

[0554] 1H NMR (300 MHz, CDCl3) δ 7.70 – 7.60 (m, 1H), 7.56 (s, 1H), 7.45 (s,1H), 7.05 – 6.88 (m, 2H), 4.83 (d, J = 13.3 Hz, 1H), 4.67 (d, J = 13.4 Hz,1H), 4.58 (dd, J = 10.2, 2.7 Hz, 1H), 4.32 (q, J = 7.1 Hz, 2H), 4.07 (d, J =11.4 Hz, 1H), 3.91 (s, 3H), 3.78 (td, J = 11.5, 2.9 Hz, 1H), 3.52 (s, 3H),3.36 – 3.26 (m, 1H), 3.24 – 3.12 (m, 1H), 1.37 (t, J = 7.1 Hz, 3H).

[0555] MS m / z (ES-API+): 484.10 [M+H]+

[0556] Dissolve 41-1 (100 mg, 0.28 mmol) in DMSO (5 mL). Add DIPEA (0.2 mL) and 1-8-P2 (72 mg, 0.42 mmol), and react at 100 °C for 5 hours. Quench the reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, elute with 200%–50% PE / EA solution, and separate compound 41-P2 (70 mg).

[0557] 1H NMR (300 MHz, CDCl3) δ 7.72–7.60 (m, 1H), 7.56 (s, 1H), 7.45 (s,1H), 7.05 – 6.96 (m, 1H), 6.96 – 6.87 (m, 1H), 4.83 (d, J = 13.3 Hz, 1H), 4.67 (d, J = 13.3 Hz, 1H), 4.58 (dd, J = 10.1, 2.7 Hz, 1H), 4.32 (q, J = 7.1Hz, 2H), 4.21 (t, J = 6.7 Hz, 2H), 3.91 (s, 3H), 3.79 (td, J = 11.5, 2.8 Hz, 1H), 3.53 (s, 3H), 3.34–3.27 (m, 1H), 1.34 (d, J = 7.4 Hz, 3H).

[0558] MS m / z (ES-API+): 484.10 [M+H]+

[0559] Example 42: Synthesis of 8-(4-chloro-2-fluorophenyl)-6-((6R)-2-(1-cyclopropyl-1H-pyrazol-4-yl)-6-methylmorpholinyl)-1-ethyl-3-methylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 42)

[0560]

[0561] Dissolve 39-1 (100 mg, 0.27 mmol) in DMSO (5 mL). Add DIPEA (0.2 mL) and 42-1 (84 mg, 0.41 mmol), and react at 100 °C for 5 h. Quench the reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 200%–50% PE / EA solution to obtain compound 42 (74 mg).

[0562] 1H NMR (400 MHz, CDCl3) δ 7.62 – 7.56 (m, 1H), 7.53 (s, 2H), 7.25 (s,1H), 7.20 (dd, J = 9.7, 1.9 Hz, 1H), 4.87 (d, J = 13.2 Hz, 1H), 4.77 (d, J =13.2 Hz, 1H), 4.57 (dd, J = 10.9, 2.7 Hz, 1H), 4.31 (q, J = 7.1 Hz, 2H), 3.80 (dtd, J = 12.3, 6.2, 2.6 Hz, 1H), 3.56 (dq, J = 7.3, 3.7 Hz, 1H), 3.52 (s, 3H), 3.00 (dd, J = 13.3, 10.9 Hz, 1H), 2.77 (dd, J = 13.3, 10.6 Hz, 1H), 1.37(t, J = 7.1 Hz, 3H), 1.31 (d, J = 6.2 Hz, 3H), 1.11 (ddt, J = 7.3, 4.9, 2.9Hz, 2H), 1.03 – 0.96 (m, 2H).

[0563] MS m / z (ES-API+): 540.10 [M+H]+

[0564] Example 43: Synthesis of 6-(2-(1-(difluoromethyl)-1H-pyrazol-4-yl)-6-methylmorpholinyl)-8-(2,4-difluorophenyl)-1-ethyl-3-methylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 43)

[0565]

[0566] Step 1: Synthesis of 1-(1-(difluoromethyl)-1H-pyrazol-4-yl)ethyl-1-one (43-2)

[0567] 43-1 (2 g, 18.18 mmol) and potassium fluoride (2.1 g, 36.36 mmol) were placed in a 100 mL double-necked flask and dissolved in 30 mL of anhydrous acetonitrile under nitrogen atmosphere. Diethyl bromodifluoromethyl phosphate (4.85 g, 18.18 mmol) was then added with stirring, and the reaction was carried out at room temperature for 12 hours. After the reaction was complete, the reaction mixture was diluted with an aqueous sodium sulfite solution, extracted three times with ethyl acetate, and the organic layer was dried over anhydrous MgSO4, filtered, and concentrated. Purification was performed on a silica gel column, eluting with 1500%–50% PE / EA solution to obtain 43-2 (2.5 g).

[0568] Step 2: Synthesis of 2-bromo-1-(1-(difluoromethyl)-1H-pyrazol-4-yl)ethyl-1-one (43-3)

[0569] 43-2 (100 mg, 0.625 mmol) and pyridinium tribromide (200 mg, 0.625 mmol) were dissolved in 5 mL of a mixed solvent (DCM:EtOH = 4:1) and reacted at room temperature for 5 hours. After the reaction was complete, the reaction mixture was diluted with an aqueous sodium sulfite solution, extracted three times with ethyl acetate, and the organic layer was dried over anhydrous MgSO4, filtered, and concentrated. Purification was performed on a silica gel column, eluting with 1000%–500% PE / EA solution to give 43-3 (170 mg).

[0570] Step 3: Synthesis of 2-(benzyl(2-hydroxypropyl)amino)-1-(1-(difluoromethyl)-1H-pyrazol-4-yl)ethyl-1-one (43-4)

[0571] DMF solution (5 mL), 1-(benzylamino)prop-2-ol (117 mg, 0.71 mmol), and N,N-diisopropylethylamine (275 mg, 2.13 mmol) were added to 43-3 (170 mg, 0.71 mmol), and the mixture was reacted at room temperature for 16 hours. After the reaction was complete, the mixture was extracted three times with ethyl acetate. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. The solution was purified on a silica gel column and eluted with 300%–50% PE / EA solution to give 43-4 (100 mg).

[0572] Step 4: Synthesis of 4-benzyl-6-(1-(difluoromethyl)-1H-pyrazol-4-yl)-2-methyl-3,4-dihydro-2H-1,4-oxazine (43-5)

[0573] Under nitrogen atmosphere and at 0°C, TES (272 mg, 1.54 mmol) and TMSOTf (343 mg, 1.54 mmol) were added to a 5 mL solution of DCM containing 43-4 (100 mg, 0.31 mmol). After the addition was complete, the mixture was heated to room temperature and reacted for 12 hours. After the reaction was complete, the reaction was quenched with water, and the mixture was extracted three times with dichloromethane. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. The solution was purified on a silica gel column and eluted with 500%–300% PE / EA solution to obtain 43-5 (50 mg).

[0574] Step 5: Synthesis of 2-(1-(difluoromethyl)-1H-pyrazol-4-yl)-6-methylmorpholine (43-6)

[0575] Dissolve 43-5 (750 mg, 2.46 mmol) in methanol (10 mL). Add 10% palladium on carbon (260 mg, 2.46 mmol) and react at 50 °C for 16 hours under a hydrogen atmosphere. After the reaction is complete, dilute with methanol, filter through diatomaceous earth, dry with anhydrous MgSO4, and concentrate to obtain 43-6 (500 mg). No further purification is required; it can be used directly in the next step.

[0576] Step 6: Synthesis of 6-(2-(1-(difluoromethyl)-1H-pyrazol-4-yl)-6-methylmorpholinyl)-8-(2,4-difluorophenyl)-1-ethyl-3-methylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 43)

[0577] Dissolve 43-6(1H,3H)-dione (100 mg, 0.28 mmol) in DMSO (5 mL). Add DIPEA (0.2 mL) and (2-(1-(difluoromethyl)-1H-pyrazol-4-yl)-6-methylmorpholine (91 mg, 0.42 mmol), and react at 100 °C for 5 h. Quench the reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, elute with 200%–20% PE / EA solution, and obtain compound 43 (54 mg).

[0578] 1 H NMR (300 MHz, CDCl3) δ 7.90 (s, 1H), 7.74 (s, 1H), 7.71 – 7.59 (m,1H), 7.07 – 6.86 (m, 3H), 4.95 (d, J = 13.3 Hz, 1H), 4.81 (dd, J = 13.3, 2.4Hz, 1H), 4.65 (dd, J = 10.9, 2.7 Hz, 1H), 4.32 (q, J = 7.1 Hz, 2H), 3.89 –3.75 (m, 1H), 3.53 (s, 3H), 2.99 (dd, J = 13.2, 10.9 Hz, 1H), 2.79 (dd, J =13.3, 10.6 Hz, 1H), 1.37 (t, J = 7.1 Hz, 3H), 1.33 (d, J = 6.2 Hz, 3H).

[0579] MS m / z (ES-API+): 534.10 [M+H]+

[0580] Example 44: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-(5-(1-methyl-1H-pyrazol-4-yl)-4-oxa-7-azaspiro[2.5]octane-7-yl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 44)

[0581]

[0582] Dissolve compound 1-7 (100 mg, 0.28 mmol) in DMSO (5 mL). Add DIPEA (0.3 mL) and 44-1 (81 mg, 0.42 mmol), and react at 100 °C for 5 hours. Quench the reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 200%–50% PE / EA solution to obtain compound 44 (64 mg).

[0583] 1 H NMR (400 MHz, CDCl3) δ 7.55 (d, J = 9.0 Hz, 1H), 7.51 (s, 1H), 7.43 (d, J = 10.0 Hz, 1H), 7.32 (dd, J = 8.3, 2.0 Hz, 1H), 7.22 (dd, J = 9.5, 1.9Hz, 1H), 4.83 (d, J = 12.2 Hz, 1H), 4.71 (dd, J = 10.9, 2.7 Hz, 1H), 4.11 (td, J = 13.6, 6.8 Hz, 1H), 3.89 (d, J = 6.9 Hz, 3H), 3.68 (d, J = 13.5 Hz,1H), 3.50 (d, J = 2.2 Hz, 3H), 3.17 (dd, J = 13.4, 10.9 Hz, 1H), 3.04 (s,3H), 1.00 (dt, J = 24.4, 7.4 Hz, 2H), 0.90 – 0.77 (m, 2H).

[0584] MS m / z (ES-API+): 512.15 [M+H]+

[0585] Example 45: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-(6-methyl-2,6-diazaspiro[3,4]octane-2-yl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 45)

[0586]

[0587] Dissolve compound 1-7 (187 mg, 0.53 mmol) in DMSO (10 mL). Add DIPEA (0.6 mL) and 45-1 (100 mg, 0.79 mmol), and react at 100 °C for 5 hours. Quench the reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 100%–20% PE / EA solution to obtain compound 45 (80 mg).

[0588] 1 H NMR (300 MHz, CDCl3) δ 7.57 (t, J = 8.0 Hz, 1H), 7.31 (dd, J = 8.4,2.0 Hz, 1H), 7.20 (dd, J = 9.6, 2.0 Hz, 1H), 4.15 (q, J = 9.0 Hz, 4H), 3.48(s, 3H), 3.01 (s, 3H), 2.76 (s, 2H), 2.62 – 2.53 (m, 2H), 2.34 (s, 3H), 2.13(t, J = 7.0 Hz, 2H).

[0589] MS m / z (ES-API+): 445.15 [M+H]+

[0590] Example 46: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-(6-methyl-2,6-diazaspiro[3,4]octane-2-yl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 46)

[0591]

[0592] Dissolve compound 1-7 (187 mg, 0.53 mmol) in DMSO (10 mL). Add DIPEA (0.6 mL) and 46-1 (100 mg, 0.79 mmol), and react at 100 °C for 5 h. Quench the reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 100%–20% PE / EA solution to obtain compound 46 (79 mg).

[0593] 1H NMR (300 MHz, CDCl3) δ 7.58 (t, J = 8.0 Hz, 1H), 7.31–7.25 (m, 1H), 7.17 (dd, J = 9.6, 2.0 Hz, 1H), 3.67 (d, J = 26.1 Hz, 2H), 3.44 (s, 3H), 3.28–3.18 (m, 2H), 3.13 (d, J = 7.1 Hz, 2H), 2.98 (s, 3H), 2.57 (s, 2H), 2.29(s, 3H), 2.15 (t, J = 6.9 Hz, 2H).

[0594] MS m / z (ES-API+): 445.15 [M+H]+

[0595] Example 47: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-(8-methyl-2,3,4a,5,6,7,8,10b-octahydro-4H-pyrazolo[3',4':6,7]cycloheptano[1,2-b][1,4]oxazin-4-yl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 47)

[0596]

[0597] Step 1: Synthesis of 1-methyl-5,6,7,8-tetrahydrocycloheptan[c]pyrazole-4(1H)-one (47-2)

[0598] 47-1 (1 g, 7.94 mmol) was dissolved in 10 mL of DMF-DMA and reacted at 100 °C for 1 hour. After the reaction was complete, the solution was concentrated under reduced pressure to remove excess DMF-DMA. 20 mL of methanol was added to the flask, and methylhydrazine dihydrochloride (1.04 g, 8.73 mmol) was added at 0 °C. The reaction was carried out at room temperature for 1.5 hours. After the reaction was complete, 50 mL of water was added to quench the reaction, and the mixture was extracted three times with dichloromethane. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. The solution was purified on a silica gel column and eluted with 500%–100% PE / EA solution to obtain 47-2 (770 mg).

[0599] Step 2: Synthesis of 5-bromo-1-methyl-5,6,7,8-tetrahydrocycloheptan[c]pyrazole-4(1H)-one (47-3)

[0600] 47-2 (770 mg, 4.70 mmol) was dissolved in 20 mL of dichloromethane and 5 mL of ethanol, and pyridine tribromide (1.5 g, 4.70 mmol) was added. The mixture was reacted at room temperature for 12 hours. After the reaction was complete, the reaction was quenched with an aqueous sodium thiosulfate solution, and the mixture was extracted three times with dichloromethane. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated to obtain 47-3 (1.1 g), which could be used directly in the next step without further purification.

[0601] Step 3: Synthesis of 5-(benzyl(2-hydroxyethyl)amino)-1-methyl-5,6,7,8-tetrahydrocyclohepta[c]pyrazole-4(1H)-one (47-4)

[0602] 47-3 (1.1 g, 4.53 mmol), 2-(benzylamino)ethanol (684 mg, 4.53 mmol), and N,N-diisopropylethylamine (1.75 g, 13.58 mmol) were dissolved in 10 mL of DMF and reacted at room temperature for 12 hours. After the reaction was complete, the mixture was diluted with water, extracted three times with ethyl acetate, and the organic layer was dried over anhydrous MgSO4, filtered, and concentrated. Purification was performed on a silica gel column, eluting with 300%–50% PE / EA solution to give 47-4 (920 mg).

[0603] Step 4: Synthesis of 5-(benzyl(2-hydroxyethyl)amino)-1-methyl-1,4,5,6,7,8-hexahydrocycloheptan[c]pyrazole-4-ol (47-5)

[0604] Add 5 mL of methanol solution to 47-4 (200 mg, 0.64 mmol), then add sodium borohydride (73 mg, 1.91 mmol) in an ice bath and react at 0°C for 1 hour. After the reaction is complete, quench the reaction with saturated ammonium chloride solution, extract three times with dichloromethane, dry the organic layer with anhydrous MgSO4, filter and concentrate to obtain 47-5 (200 mg), which can be used directly in the next step without purification.

[0605] Step 5: Synthesis of 4-benzyl-8-methyl-3,4,4a,5,6,7,8,10b-octahydro-2H-pyrazolo[3',4':6,7]cycloheptano[1,2-b][1,4]oxazine (47-6)

[0606] 10 mL of 1,4-dioxane was added to 47-5 (200 mg, 0.63 mmol) to dissolve it, followed by the slow addition of 0.45 mL of concentrated hydrochloric acid and 0.45 mL of water. The reaction was carried out at 110 °C for 2 hours. After the reaction was complete, the reaction was quenched with sodium carbonate solution, and the mixture was extracted three times with ethyl acetate. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. The solution was purified on a silica gel column and eluted with 300%–20% PE / EA solution to obtain 47-6 (164 mg).

[0607] Step 6: Synthesis of 8-methyl-3,4,4a,5,6,7,8,10b-octahydro-2H-pyrazolo[3',4':6,7]cycloheptano[1,2-b][1,4]oxazine (47-7)

[0608] Dissolve 47-6 (164 mg, 0.55 mmol) in methanol (3 mL). Add 10% palladium on carbon (60 mg, 0.55 mmol) and react at 50 °C for 24 hours under a hydrogen atmosphere. After the reaction is complete, dilute with methanol, filter through diatomaceous earth, dry with anhydrous MgSO4, and concentrate to obtain 47-7 (110 mg), which can be used directly for the next step without further purification.

[0609] Step 7: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-(8-methyl-2,3,4a,5,6,7,8,10b-octahydro-4H-pyrazolo[3',4':6,7]cycloheptano[1,2-b][1,4]oxazin-4-yl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 47)

[0610] Dissolve compound 1-7 (94 mg, 0.27 mmol) in DMSO (5 mL). Add DIPEA (136 μL, 0.79 mmol) and 47-7 (110 mg, 0.53 mmol), and react at 100 °C for 5 hours. Quench the crude reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 300%–20% PE / EA solution to obtain compound 47 (108 mg).

[0611] 1H NMR (300 MHz, CDCl3) δ 7.66 – 7.48 (m, 1H), 7.40 (s, 1H), 7.30 (d,J = 9.2 Hz, 1H), 7.21 (dd, J = 9.7, 1.9 Hz, 1H), 4.70 (s, 1H), 4.56 (s, 1H), 4.09 (d, J = 10.5 Hz, 1H), 3.79 (d, J = 8.2 Hz, 4H), 3.50 (s, 3H), 3.40 (s,1H), 3.04 (s, 3H), 2.87 (d, J = 16.2 Hz, 1H), 2.79 – 2.64 (m, 1H), 2.55 (q, J= 12.2 Hz, 1H), 2.10 (s, 1H), 1.96 – 1.60 (m, 3H).

[0612] MS m / z (ES-API+): 526.20 [M+H]+

[0613] Example 48: Synthesis of 6-(5-(1-cyclopropyl-1H-pyrazol-4-yl)-4-oxa-7-azaspiro[2.5]octane-7-yl)-8-(2,4-difluorophenyl)-1,3-dimethylpyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 48)

[0614]

[0615] Step 1: Synthesis of 2-(benzyl((1-hydroxycyclopropyl)methyl)amino)-1-(1-cyclopropyl-1H-pyrazol-4-yl)acetone (48-2)

[0616] 1-[(benzylamino)methyl]cyclopropanol (2 g, 11.3 mmol) and 48-1 (2 g, 8.8 mmol) were dissolved in CH3CN (50 mL). DIPEA (3.9 mL, 22.6 mmol) was added, and the mixture was heated and stirred at 70 °C for 4 h. After the reaction was complete, the mixture was extracted three times with EA. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. Purification was performed on a silica gel column, eluting with 500%–100% PE / EA solution to give 48-2 (2.5 g).

[0617] Step 2: Synthesis of 7-benzyl-5-(1-cyclopropyl-1H-pyrazol-4-yl)-4-oxa-7-azaspiro[2.5]octane (48-3)

[0618] 48-2 (2.5 g, 7.7 mmol) was dissolved in anhydrous DCM and purged with nitrogen. Trimethylsilane (11.7 mL, 76.9 mmol) was added under ice bath conditions and the mixture was stirred for 15 min. Triethylsiloxane trifluoromethanesulfonate (20 mL, 115.3 mmol) was slowly added under ice bath conditions. The reaction was allowed to proceed at room temperature for 10 h. After the reaction was complete, the reaction solution was quenched with NaHCO3. The mixture was extracted three times with DCM, and the organic layer was dried over anhydrous MgSO4, filtered, and concentrated to obtain 48-3 (2.3 g).

[0619] Step 3: Synthesis of 5-(1-cyclopropyl-1H-pyrazol-4-yl)-4-oxa-7-azaspiro[2.5]octane (48-4)

[0620] 48-3 (2.3 g, 7.4 mmol) was dissolved in MeOH, and Pd / C (1.1 g) was added. The H2 in the reaction solution was removed by vacuum, and the reaction was carried out at 80 °C for 10 h. 48-4 (2 g) was obtained.

[0621] Step 4: Synthesis of 6-(5-(1-cyclopropyl-1H-pyrazol-4-yl)-4-oxa-7-azaspiro[2.5]octane-7-yl)-8-(2,4-difluorophenyl)-1,3-dimethylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 48)

[0622] Dissolve 8-5 (70 mg, 0.21 mmol) in DMSO (5 mL). Add DIPEA (0.1 mL) and 48-4 (100 mg, 0.46 mmol), and react at 100 °C for 5 h. Quench the crude reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 100%–20% PE / EA solution to obtain compound 48 (60 mg).

[0623] 1H NMR (300 MHz, CDCl3) δ 7.66 – 7.58 (m, 1H), 7.54 (d, J = 5.8 Hz, 2H), 7.07 (t, J = 8.5 Hz, 1H), 6.94 (ddd, J = 10.5, 8.6, 2.3 Hz, 1H), 4.70(d, J = 10.5 Hz, 1H), 4.17 – 4.04 (m, 2H), 3.68 (d, J = 13.4 Hz, 1H), 3.64 –3.55 (m, 1H), 3.51 (s, 3H), 3.22 – 3.10 (m, 1H), 3.03 (s, 3H), 1.29 – 1.21(m, 2H), 1.12 (d, J = 3.8 Hz, 2H), 1.04 (q, J = 5.5 Hz, 2H), 0.79 (d, J = 5.8Hz, 2H).

[0624] MS m / z (ES-API+): 522.10 [M+H] +

[0625] Example 49: Synthesis of 6-(2-(1-cyclopropyl-1H-pyrazol-4-yl)morpholinyl)-8-(2-fluoro-4-(trifluoromethyl)phenyl)-1,3-dimethylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 49)

[0626]

[0627] Step 1: Synthesis of 2-(benzyl(2-hydroxyethyl)amino)-1-(1-cyclopropyl-1H-pyrazol-4-yl)acetone (49-1)

[0628] 2-(benzylamino)ethane-1-ol (1.5 g, 9.9 mmol) and 48-1 (2 g, 8.8 mmol) were dissolved in CH3CN (50 mL). DIPEA (3.9 mL, 22.6 mmol) was added, and the mixture was heated and stirred at 70 °C for 4 h. After the reaction was complete, the mixture was extracted three times with EA. The organic layer was dried over anhydrous MgSO4, filtered, and concentrated. Purification was performed on a silica gel column, eluting with 500%–100% PE / EA solution to give 49-1 (2.5 g).

[0629] Step 2: Synthesis of 4-benzyl-2-(1-cyclopropyl-1H-pyrazol-4-yl)morpholine (49-2)

[0630] 49-1 (2.5 g, 8.4 mmol) was dissolved in anhydrous DCM and evacuated under nitrogen. Trimethylsilane (11.7 mL, 76.9 mmol) was added under ice bath conditions and the mixture was stirred for 15 min. Triethylsiloxane trifluoromethanesulfonate (20 mL, 115.3 mmol) was slowly added under ice bath conditions. The reaction was allowed to proceed at room temperature for 10 h. After the reaction was complete, the reaction solution was quenched with NaHCO3. The mixture was extracted three times with DCM, and the organic layer was dried over anhydrous MgSO4, filtered, and concentrated to obtain 49-2 (2.3 g).

[0631] Step 3: Synthesis of 2-(1-cyclopropyl-1H-pyrazol-4-yl)morpholine (49-3)

[0632] 49-2 (2.3 g, 9.7 mmol) was dissolved in MeOH, and Pd / C (1.1 g) was added. The H2 in the reaction solution was removed by vacuum, and the reaction was carried out at 80 °C for 10 h. 49-3 (2 g) was obtained.

[0633] Step 4: Synthesis of 6-(2-(1-cyclopropyl-1H-pyrazol-4-yl)morpholino)-8-(2-fluoro-4-(trifluoromethyl)phenyl)-1,3-dimethylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 49)

[0634] Dissolve compound 49-3 (100 mg, 0.26 mmol) in DMSO (5 mL). Add DIPEA (0.1 mL) and 2-(1-cyclopropyl-1H-pyrazol-4-yl)morpholine (100 mg, 0.46 mmol), and react at 100 °C for 5 h. Quench the crude reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, elute with 100%–20% PE / EA solution, and obtain compound 49 (70 mg).

[0635] 1H NMR (300 MHz, CDCl3) δ 7.77 (t, J = 7.4 Hz, 1H), 7.60 (d, J = 7.8Hz, 1H), 7.49 (d, J = 18.3 Hz, 3H), 4.71 (d, J = 13.1 Hz, 1H), 4.55 (t, J =14.6 Hz, 2H), 4.14 – 4.00 (m, 1H), 3.76 (t, J = 11.1 Hz, 1H), 3.57 (s, 1H), 3.52 (s, 3H), 3.22 (dt, J = 23.8, 11.8 Hz, 2H), 3.03 (s, 3H), 1.10 (s, 2H),1.01 (d, J = 7.1 Hz, 2H).

[0636] MS m / z (ES-API+): 546.08 [M+H]

[0637] Example 50: Synthesis of 6-(2-(1-cyclopropyl-1H-pyrazol-4-yl)-6-methylmorpholinyl)-8-(2-fluoro-4-(trifluoromethyl)phenyl)-1,3-dimethylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (Compound 50)

[0638]

[0639] Dissolve 14-3 (100 mg, 0.26 mmol) in DMSO (5 mL). Add DIPEA (0.1 mL) and 2-(1-cyclopropyl-1H-pyrazol-4-yl)-6-methylmorpholine (106 mg, 0.52 mmol), and react at 100 °C for 5 h. Quench the crude reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, elute with 100%–20% PE / EA solution, and separate to obtain compound 50 (70 mg).

[0640] 1H NMR (400 MHz, CDCl3) δ 7.77 (s, 1H), 7.61 (d, J = 7.0 Hz, 1H), 7.54(s, 2H), 7.47 (d, J = 9.4 Hz, 1H), 4.75 (d, J = 11.9 Hz, 1H), 4.67 (d, J =12.7 Hz, 1H), 4.56 (d, J = 10.5 Hz, 1H), 3.75 (dd, J = 15.2, 9.0 Hz, 2H), 3.57 (s, 1H), 3.52 (s, 3H), 3.02 (s, 3H), 2.78 (t, J = 11.9 Hz, 1H), 1.31 (d,J = 6.0 Hz, 3H), 1.12 (s, 2H), 1.02 (d, J = 6.4 Hz, 2H).

[0641] MS m / z (ES-API+): 560.10 [M+H]

[0642] Example 51: Synthesis of 6-(2-(1-cyclopropyl-1H-pyrazol-4-yl)-6-methylmorpholinyl)-8-(2,4-difluorophenyl)-1-isopropyl-3-methylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 51)

[0643]

[0644] Step 1: Synthesis of 6-chloro-8-(2,4-difluorophenyl)-1-isopropyl-3-methylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (51-1)

[0645] Dissolve 8-5 (130 mg, 0.40 mmol) in 10 mL of anhydrous DMF solution. Add iodoisopropane (0.1 mL, 1.0 mmol) and K₂CO₃ (166 mg, 1.2 mmol), and react at 80 °C for 3 hours. After the reaction is complete, quench the reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO₄, filter and concentrate. Purify on a silica gel column, elute with 800%–400% PE / EA solution, and separate to obtain 51-1 (50 mg).

[0646] Step 2: Synthesis of 6-(2-(1-cyclopropyl-1H-pyrazol-4-yl)-6-methylmorpholinyl)-8-(2,4-difluorophenyl)-1-isopropyl-3-methylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 51)

[0647] Dissolve 51-1 (50 mg, 0.14 mmol) in DMSO (5 mL). Add DIPEA (0.1 mL) and 50-1 (102 mg, 0.52 mmol), and react at 100 °C for 5 h. Quench the crude reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 100%–20% PE / EA solution to obtain compound 51 (20 mg).

[0648] 1 H NMR (300 MHz, CDCl3) δ 7.68 – 7.57 (m, 1H), 7.54 (s, 2H), 7.00 (td,J = 8.3, 2.6 Hz, 1H), 6.91 (ddd, J = 10.0, 9.0, 2.5 Hz, 1H), 5.10 (dt, J =12.5, 6.2 Hz, 1H), 4.88 (d, J = 13.1 Hz, 1H), 4.78 (d, J = 13.2 Hz, 1H), 4.58(dd, J = 10.9, 2.7 Hz, 1H), 3.81 (ddd, J = 10.6, 6.2, 2.6 Hz, 1H), 3.56 (tt,J = 7.3, 3.8 Hz, 1H), 3.50 (s, 3H), 3.00 (dd, J = 13.3, 10.9 Hz, 1H), 2.77(dd, J = 13.3, 10.6 Hz, 1H), 1.34 (s, 3H), 1.32 (s, 3H), 1.24 (d, J = 7.3 Hz, 3H), 1.11 (qd, J = 5.0, 3.3 Hz, 2H), 1.06 – 0.96 (m, 2H).

[0649] MS m / z (ES-API+): 538.13 [M+H]

[0650] Example 52: Synthesis of 6-(2-(1-cyclopropyl-1H-pyrazol-4-yl)-6-methylmorpholinyl)-1-ethyl-8-(2-fluoro-4-(trifluoromethyl)phenyl)-3-methylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 52)

[0651]

[0652] Dissolve 27-1 (130 mg, 0.32 mmol) in DMSO (5 mL). Add DIPEA (0.1 mL) and 50-1 (130 mg, 0.63 mmol), and react at 100 °C for 5 h. Quench the crude reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, elute with 100%–20% PE / EA solution, and separate to obtain compound 52 (40 mg).

[0653] 1 H NMR (300 MHz, CDCl3) δ 7.75 (t, J = 7.4 Hz, 1H), 7.55 (d, J = 1.5Hz, 1H), 7.53 (s, 2H), 7.49 – 7.41 (m, 1H), 4.87 (d, J = 13.0 Hz, 1H), 4.77(d, J = 13.2 Hz, 1H), 4.57 (dd, J = 10.9, 2.7 Hz, 1H), 4.30 (q, J = 7.1 Hz,2H), 3.80 (ddt, J = 12.5, 6.3, 3.2 Hz, 1H), 3.57 (dt, J = 7.3, 3.8 Hz, 1H),3.52 (s, 3H), 3.01 (dd, J = 13.3, 10.9 Hz, 1H), 2.78 (dd, J = 13.3, 10.6 Hz,1H), 1.36 (t, J = 7.1 Hz, 3H), 1.31 (d, J = 6.2 Hz, 3H), 1.14 – 1.06 (m, 2H),1.04 – 0.94 (m, 2H).

[0654] MS m / z (ES-API+): 538.11 [M+H] +

[0655] Example 53: Synthesis of 6-(2-(1-cyclopropyl-1H-pyrazol-4-yl)morpholino)-8-(2,4-difluorophenyl)-1-ethyl-3-methylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 53)

[0656]

[0657] Dissolve 41-1 (160 mg, 0.45 mmol) in DMSO (5 mL). Add DIPEA (0.1 mL) and 49-3 (175 mg, 0.91 mmol), and react at 100 °C for 5 h. Quench the crude reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, elute with 100%–20% PE / EA solution, and obtain compound 53 (50 mg).

[0658] 1 H NMR (300 MHz, CDCl3) δ 7.64 (td, J = 8.3, 6.6 Hz, 1H), 7.52 (d, J =2.6 Hz, 2H), 7.05 – 6.96 (m, 1H), 6.92 (ddd, J = 10.1, 8.9, 2.4 Hz, 1H), 4.88– 4.77 (m, 1H), 4.68 (d, J = 13.6 Hz, 1H), 4.55 (dd, J = 10.3, 2.8 Hz, 1H), 4.32 (q, J = 7.1 Hz, 2H), 4.07 (ddd, J = 11.6, 3.6, 1.7 Hz, 1H), 3.77 (td, J= 11.5, 2.8 Hz, 1H), 3.57 (dq, J = 7.3, 3.7 Hz, 1H), 3.52 (s, 3H), 3.33 –3.22 (m, 1H), 3.17 (dd, J = 13.4, 10.4 Hz, 1H), 1.36 (t, J = 7.1 Hz, 3H), 1.15 – 1.07 (m, 2H), 1.06 – 0.95 (m, 2H).

[0659] MS m / z (ES-API+): 510.10 [M+H]

[0660] Example 54: Synthesis of 1-ethyl-8-(2-fluoro-4-(trifluoromethyl)phenyl)-3-methyl-6-(5-(1-methyl-1H-pyrazol-4-yl)-4-oxa-7-azaspiro[2.5]octane-7-yl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 54)

[0661]

[0662] Dissolve 27-1 (100 mg, 0.25 mmol) in DMSO (5 mL). Add DIPEA (0.1 mL) and 44-1 (100 mg, 0.52 mmol), and react at 100 °C for 5 h. Quench the crude reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 100%–20% PE / EA solution to obtain compound 54 (40 mg).

[0663] 1 H NMR (400 MHz, CDCl3) δ 7.74 (t, J = 7.4 Hz, 1H), 7.55 (s, 1H), 7.52(s, 1H), 7.45 (s, 1H), 7.43 (d, J = 2.5 Hz, 1H), 4.95 (d, J = 13.4 Hz, 1H), 4.74 (dd, J = 10.9, 2.7 Hz, 1H), 4.30 (q, J = 7.1 Hz, 2H), 4.19 (d, J = 13.3Hz, 1H), 3.88 (s, 3H), 3.69 (d, J = 13.3 Hz, 1H), 3.52 (d, J = 2.2 Hz, 3H),3.19 (dd, J = 13.4, 10.8 Hz, 1H), 1.36 (t, J = 7.0 Hz, 3H), 1.09 – 0.91 (m,2H), 0.78 (p, J = 7.5 Hz, 2H).

[0664] MS m / z (ES-API+): 560.10 [M+H]

[0665] Example 55: Synthesis of 6-(2-(1-(difluoromethyl)-1H-pyrazol-4-yl)-6-methylmorpholinyl)-1-ethyl-8-(2-fluoro-4-(trifluoromethyl)phenyl)-3-methylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 55)

[0666]

[0667] Dissolve 27-1 (100 mg, 0.25 mmol) in DMSO (5 mL). Add DIPEA (0.1 mL) and 43-6 (110 mg, 0.51 mmol), and react at 100 °C for 5 h. Quench the crude reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 100%–20% PE / EA solution to obtain compound 55 (30 mg).

[0668] 1 H NMR (300 MHz, CDCl3) δ 7.90 (s, 1H), 7.77 (d, J = 7.4 Hz, 1H), 7.74(s, 1H), 7.59 – 7.51 (m, 1H), 7.45 (d, J = 9.7 Hz, 1H), 4.94 (d, J = 13.2 Hz, 1H), 4.80 (d, J = 13.1 Hz, 1H), 4.65 (d, J = 10.1 Hz, 1H), 4.31 (q, J = 7.1Hz, 2H), 3.83 (t, J = 8.6 Hz, 1H), 3.53 (s, 3H), 3.00 (dd, J = 13.1, 10.9 Hz, 1H), 2.80 (dd, J = 13.2, 10.6 Hz, 1H), 1.35 – 1.31 (m, 3H), 1.26 (tt, J =5.3, 2.5 Hz, 3H).

[0669] MS m / z (ES-API+): 584.18 [M+H]

[0670] Example 56: Synthesis of 6-((6R)-2-(1-cyclopropyl-1H-pyrazol-4-yl)-6-methylmorpholinyl)-8-(2,4-difluorophenyl)-1-ethyl-3-methylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 56)

[0671]

[0672] Dissolve 41-1 (100 mg, 0.28 mmol) in DMSO (5 mL). Add DIPEA (0.1 mL) and 42-1 (120 mg, 0.58 mmol), and react at 100 °C for 5 h. Quench the crude reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, elute with 100%–20% PE / EA solution, to obtain compound 56 (40 mg).

[0673] 1 H NMR (400 MHz, CDCl3) δ 7.63 (tt, J = 8.4, 6.2 Hz, 1H), 7.53 (d, J =2.4 Hz, 2H), 7.05 – 6.96 (m, 1H), 6.95 – 6.87 (m, 1H), 4.88 (d, J = 13.5 Hz, 1H), 4.77 (d, J = 13.2 Hz, 1H), 4.57 (dd, J = 10.9, 2.7 Hz, 1H), 4.31 (q, J =7.1 Hz, 2H), 3.79 (dtd, J = 12.5, 6.3, 2.6 Hz, 1H), 3.55 (dq, J = 7.3, 3.7Hz, 1H), 3.51 (s, 3H), 2.99 (dd, J = 13.3, 10.9 Hz, 1H), 2.76 (dd, J = 13.3,10.6 Hz, 1H), 1.36 (td, J = 7.1, 2.1 Hz, 3H), 1.30 (d, J = 6.3 Hz, 3H), 1.13– 1.05 (m, 2H), 1.02 – 0.96 (m, 2H).

[0674] MS m / z (ES-API+): 524.11 [M+H]

[0675] Example 57: Synthesis of 8-(4-chloro-2-fluorophenyl)-6-(2-(1-cyclopropyl-1H-pyrazol-4-yl)morpholinyl)-1-ethyl-3-methylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 57)

[0676]

[0677] Dissolve 39-1 (100 mg, 0.27 mmol) in DMSO (5 mL). Add DIPEA (0.1 mL) and 49-3 (100 mg, 0.52 mmol), and react at 100 °C for 5 h. Quench the crude reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 100%–20% PE / EA solution to obtain compound 57 (40 mg).

[0678] 1 H NMR (400 MHz, CDCl3) δ 7.59 (t, J = 7.8 Hz, 1H), 7.52 (d, J = 3.0Hz, 2H), 7.27 (d, J = 2.0 Hz, 1H), 7.20 (dd, J = 9.7, 2.0 Hz, 1H), 4.82 (d, J= 13.3 Hz, 1H), 4.67 (d, J = 13.3 Hz, 1H), 4.54 (dd, J = 10.4, 2.8 Hz, 1H), 4.31 (q, J = 7.1 Hz, 2H), 4.06 (ddd, J = 11.5, 3.6, 1.7 Hz, 1H), 3.77 (td, J= 11.5, 2.8 Hz, 1H), 3.57 (dq, J = 7.3, 3.7 Hz, 1H), 3.52 (s, 3H), 3.32 –3.21 (m, 1H), 3.17 (dd, J = 13.4, 10.4 Hz, 1H), 1.37 (t, J = 7.1 Hz, 3H), 1.12 – 1.06 (m, 2H), 1.00 (ddd, J = 7.4, 4.5, 2.8 Hz, 2H).

[0679] MS m / z (ES-API+): 526.07 [M+H]

[0680] Example 58: Synthesis of 8-(2,4-difluorophenyl)-1,3-dimethyl-6-((2R)-2-methyl-6-(1-methyl-1H-pyrazol-4-yl)morpholinyl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 58)

[0681]

[0682] Dissolve compound 8-6 (100 mg, 0.30 mmol) in DMSO (3 mL). Add DIPEA (0.15 mL) and 3-7 (85 mg, 0.44 mmol), and react at 100 °C for 5 hours. Quench the crude reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 100%–20% PE / EA solution to obtain compound 58 (85 mg).

[0683] 1H NMR (400 MHz, CDCl3) δ 7.63 (td, J = 8.3, 6.2 Hz, 1H), 7.55 (s,1H), 7.46 (s, 1H), 7.07 (td, J = 8.3, 2.5 Hz, 1H), 6.94 (ddd, J = 10.9, 8.7,2.5 Hz, 1H), 4.73 (dd, J = 35.5, 13.2 Hz, 2H), 4.58 (dd, J = 10.9, 2.6 Hz,1H), 3.91 (s, 3H), 3.79 (s, 1H), 3.51 (s, 3H), 3.03 (s, 3H), 3.03 – 2.95 (m,1H), 2.78 (dd, J = 13.2, 10.6 Hz, 1H), 1.31 (d, J = 6.2 Hz, 3H).

[0684] MS m / z (ES-API+): 484.20 [M+H]+

[0685] Example 59: Synthesis of 6-(2-(1-cyclopropyl-1H-pyrazol-4-yl)morpholino)-8-(2,4-difluorophenyl)-1,3-dimethylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 59)

[0686]

[0687] Dissolve 8-6 (70 mg, 0.21 mmol) in DMSO (3 mL). Add DIPEA (0.1 mL) and 49-3 (100 mg, 0.51 mmol), and react at 100 °C for 5 hours. Quench the crude reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 100%–20% PE / EA solution to obtain compound 59 (28 mg).

[0688] 1H NMR (300 MHz, CDCl3) δ 7.63 (d, J = 7.6 Hz, 1H), 7.51 (s, 2H), 7.07 (t, J = 8.3 Hz, 1H), 6.94 (t, J = 9.2 Hz, 1H), 4.78 – 4.57 (m, 1H), 4.52(d, J = 12.0 Hz, 1H), 4.07 (d, J = 11.8 Hz, 1H), 3.75 (t, J = 11.6 Hz, 1H), 3.58 (d, J = 11.9 Hz, 1H), 3.55 – 3.44 (m, 3H), 3.22 (dt, J = 22.6, 10.9 Hz, 2H), 3.04 (s, 3H), 1.27 (d, J = 8.8 Hz, 1H), 1.10 (s, 2H), 1.00 (d, J = 7.8Hz, 2H).

[0689] MS m / z (ES-API+): 496.20 [M+H]+

[0690] Example 60: Synthesis of 6-(2-(1-(difluoromethyl)-1H-pyrazol-4-yl)-6-methylmorpholinyl)-8-(2-fluoro-4-(trifluoromethyl)phenyl)-1,3-dimethylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 60)

[0691]

[0692] Dissolve 14-3 (100 mg, 0.26 mmol) in DMSO (5 mL). Add DIPEA (0.14 mL) and 43-6 (84 mg, 0.39 mmol), and react at 100 °C for 5 hours. Quench the crude reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, elute with 500%–100% PE / EA solution, and separate to give compound 60 (160 mg).

[0693] 1H NMR (300 MHz, CDCl3) δ 7.89 (s, 1H), 7.78 (t, J = 7.4 Hz, 1H), 7.72 (s, 1H), 7.68 – 7.58 (m, 2H), 7.48 (dd, J = 9.6, 1.5 Hz, 1H), 4.81 (d, J= 13.3 Hz, 1H), 4.70 (d, J = 13.4 Hz, 1H), 4.63 (d, J = 10.8 Hz, 1H), 3.82(s, 1H), 3.52 (s, 3H), 3.03 (s, 3H), 3.02 – 2.94 (m, 1H), 2.79 (dd, J = 13.3,10.6 Hz, 1H), 1.33 (d, J = 6.2 Hz, 3H).

[0694] MS m / z (ES-API+): 570.20 [M+H]+

[0695] Example 61: Synthesis of 8-(4-chloro-2-fluorophenyl)-6-(2-(1-(difluoromethyl)-1H-pyrazol-4-yl)-6-methylmorpholinyl)-1,3-dimethylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 61)

[0696]

[0697] Dissolve compound 1-7 (100 mg, 0.28 mmol) in DMSO (5 mL). Add DIPEA (0.14 mL) and 43-6 (122 mg, 0.56 mmol), and react at 100 °C for 5 hours. Quench the crude reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, elute with 500%–100% PE / EA solution, and separate to obtain compound 61 (120 mg).

[0698] 1H NMR (300 MHz, CDCl3) δ 7.89 (s, 1H), 7.72 (s, 1H), 7.58 (t, J =8.0 Hz, 1H), 7.42 – 7.29 (m, 2H), 7.24 (dd, J = 9.5, 1.9 Hz, 1H), 4.76 (dd, J= 34.9, 13.2 Hz, 2H), 4.66 – 4.58 (m, 1H), 3.80 (d, J = 7.5 Hz, 1H), 3.51 (s,3H), 3.05 (s, 3H), 2.99 (dd, J = 13.2, 10.9 Hz, 1H), 2.79 (dd, J = 13.3, 10.6Hz, 1H), 1.33 (d, J = 6.2 Hz, 3H).

[0699] MS m / z (ES-API+): 536.20 [M+H]+

[0700] Example 62: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-(2-methyl-6-(1-(oxecyclobutan-3-yl)-1H-pyrazol-4-yl)morpholinyl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 62)

[0701]

[0702] Dissolve compound 1-7 (100 mg, 0.26 mmol) in DMSO (5 mL). Add DIPEA (0.14 mL) and 62-1 (120 mg, 0.54 mmol), and react at 100 °C for 5 hours. Quench the crude reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 300%–50% PE / EA solution to obtain compound 62 (130 mg).

[0703] 1H NMR (300 MHz, CDCl3) δ 7.68 (s, 1H), 7.65 (s, 1H), 7.58 (t, J =7.8 Hz, 2H), 7.33 (d, J = 8.0 Hz, 1H), 5.43 (q, J = 6.8 Hz, 1H), 5.04 (d, J =7.1 Hz, 4H), 4.98 (d, J = 5.6 Hz, 1H), 4.74 (dd, J = 32.4, 13.2 Hz, 2H), 4.60 (d, J = 10.3 Hz, 1H), 3.79 (s, 1H), 3.51 (s, 3H), 3.04 (s, 3H), 2.85 – 2.71(m, 1H), 1.32 (d, J = 6.1 Hz, 3H).

[0704] MS m / z (ES-API+): 542.20 [M+H]+

[0705] Example 63: Synthesis of 8-(4-chloro-2-fluorophenyl)-6-(2-(1-cyclopropyl-1H-pyrazol-4-yl)-6-methylmorpholinyl)-1,3-dimethylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 63)

[0706]

[0707] Dissolve compound 1-7 (114 mg, 0.32 mmol) in DMSO (4 mL). Add DIPEA (0.17 mL) and 50-1 (100 mg, 0.48 mmol), and react at 100 °C for 5 hours. Quench the crude reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 300%–30% PE / EA solution to obtain compound 63 (100 mg).

[0708] 1H NMR (300 MHz, CDCl3) δ 7.58 (t, J = 8.0 Hz, 1H), 7.52 (s, 2H), 7.33 (dd, J = 8.4, 2.0 Hz, 1H), 7.22 (dd, J = 9.6, 1.9 Hz, 1H), 4.71 (dd, J =24.4, 13.2 Hz, 2H), 4.55 (dd, J = 10.9, 2.7 Hz, 1H), 3.76 (d, J = 8.3 Hz,1H), 3.57 (dq, J = 7.4, 3.6 Hz, 1H), 3.50 (s, 3H), 3.04 (s, 3H), 3.03 – 2.94(m, 1H), 2.77 (dd, J = 13.2, 10.6 Hz, 1H), 1.31 (d, J = 6.2 Hz, 3H), 1.15 –1.05 (m, 2H), 1.01 (dtd, J = 7.5, 5.6, 3.6 Hz, 2H).

[0709] MS m / z (ES-API+): 526.20 [M+H]+

[0710] Example 64: Synthesis of 8-(4-chloro-2-fluorophenyl)-6-(2-(1-cyclopropyl-1H-pyrazol-4-yl)morpholinyl)-1,3-dimethylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 64)

[0711]

[0712] Dissolve compound 1-7 (100 mg, 0.28 mmol) in DMSO (4 mL). Add DIPEA (0.14 mL) and 49-3 (108 mg, 0.56 mmol), and react at 100 °C for 5 hours. Quench the crude reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 300%–40% PE / EA solution to obtain compound 64 (90 mg).

[0713] 1H NMR (300 MHz, CDCl3) δ 7.58 (t, J = 8.0 Hz, 1H), 7.51 (s, 2H), 7.32 (ddd, J = 8.3, 2.0, 0.6 Hz, 1H), 7.22 (dd, J = 9.6, 2.0 Hz, 1H), 4.71(d, J = 13.4 Hz, 1H), 4.64 – 4.44 (m, 2H), 4.12 – 4.01 (m, 1H), 3.75 (td, J =11.6, 2.8 Hz, 1H), 3.57 (tt, J = 7.3, 3.9 Hz, 1H), 3.50 (s, 3H), 3.33 – 3.10(m, 2H), 3.04 (s, 3H), 1.16 – 1.05 (m, 2H), 1.06 – 0.96 (m, 2H).

[0714] MS m / z (ES-API+): 512.20 [M+H]+

[0715] Example 65: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-(2-methyl-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)morpholino)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 65)

[0716]

[0717] Step 1: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-(2-methyl-6-(1H-pyrazol-4-yl)morpholinyl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (65-2)

[0718] Dissolve 1-7 (460 mg, 1.46 mmol) in DMSO (10 mL). Add DIPEA (0.82 mL) and 65-1 (368 mg, 2.19 mmol), and react at 100 °C for 5 hours. Quench the crude reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, elute with 300%–20% PE / EA solution, and separate to obtain 65-2 (600 mg).

[0719] Step 1: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-(2-methyl-6-(1-(2,2,2-trifluoroethyl)-1H-pyrazol-4-yl)morpholino)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 65)

[0720] Dissolve 65-2 (100 mg, 0.21 mmol) in 5 mL of DMF. Add cesium carbonate (135 mg, 0.41 mmol) and 2,2,2-trifluoroethyltrifluoromethanesulfonate (60 mg, 0.26 mmol), and react at room temperature for 5 hours. Quench the crude reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 300%–100% PE / EA solution to obtain compound 65 (116 mg).

[0721] 1 H NMR (300 MHz, CDCl3) δ 7.60 (ddd, J = 19.3, 12.8, 7.1 Hz, 3H), 7.39– 7.29 (m, 1H), 7.22 (s, 1H), 4.69 (ddt, J = 24.8, 19.3, 11.8 Hz, 4H), 3.80(s, 2H), 3.51 (d, J = 2.7 Hz, 3H), 3.05 (d, J = 2.9 Hz, 4H), 2.78 (t, J =12.0 Hz, 1H), 1.32 (dd, J = 6.4, 2.6 Hz, 2H), 1.24 (dd, J = 6.6, 2.6 Hz, 1H).

[0722] MS m / z (ES-API+): 568.20 [M+H]+

[0723] Example 66: Synthesis of 8-(4-chloro-2-fluorophenyl)-1,3-dimethyl-6-(2-methyl-6-(1-(tetrahydrofuran-3-yl)-1H-pyrazol-4-yl)morpholinyl)pyrimido[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 66)

[0724]

[0725] Dissolve 65-2 (100 mg, 0.21 mmol) in 5 mL of DMF. Add cesium carbonate (135 mg, 0.41 mmol) and 3-iodotetrahydrofuran (60 mg, 0.31 mmol), and react at 100 °C for 4 hours. Quench the crude reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 300%–10% PE / EA solution to obtain compound 66 (96 mg).

[0726] 1 H NMR (400 MHz, CDCl3) δ 7.64 – 7.52 (m, 2H), 7.47 (d, J = 3.3 Hz, 1H), 7.33 (dt, J = 8.4, 2.6 Hz, 1H), 7.23 (dt, J = 9.5, 2.4 Hz, 1H), 5.07 –4.91 (m, 1H), 4.76 (d, J = 13.0 Hz, 1H), 4.67 (d, J = 13.6 Hz, 1H), 4.58 (dt,J = 10.9, 6.2 Hz, 1H), 4.18 – 4.09 (m, 1H), 4.07 (t, J = 3.2 Hz, 1H), 4.03(d, J = 9.7 Hz, 1H), 3.99 – 3.85 (m, 2H), 3.79 (d, J = 8.2 Hz, 1H), 3.50 (s,3H), 3.04 (s, 3H), 2.78 (dd, J = 13.2, 10.7 Hz, 1H), 2.45 (dq, J = 13.1, 7.9Hz, 1H), 2.38 – 2.27 (m, 1H), 1.31 (d, J = 6.2 Hz, 3H).

[0727] MS m / z (ES-API+): 556.20 [M+H]+

[0728] Example 67: Synthesis of 8-(4-chloro-2-fluorophenyl)-6-(2-(1-isopropyl-1H-pyrazol-4-yl)-6-methylmorpholinyl)-1,3-dimethylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 67)

[0729]

[0730] Dissolve 65-2 (100 mg, 0.21 mmol) in 5 mL of DMF. Add cesium carbonate (135 mg, 0.41 mmol) and 2-iodopropane (69 mg, 0.42 mmol), and react at 80 °C for 12 hours in the dark. Quench the crude reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 300%–20% PE / EA solution to obtain compound 67 (110 mg).

[0731] 1H NMR (300 MHz, CDCl3) δ7.65–7.41 (m, 3H), 7.33 (d, J = 8.4 Hz, 1H), 7.23 (d, J = 9.2 Hz, 1H), 4.72 (dd, J = 27.4, 13.2 Hz, 1H), 4.62–4.54 (m,1H), 4.48 (p, J = 6.7 Hz, 1H), 4.40 (s, 1H), 3.78 (s, 1H), 3.51 (s, 3H), 3.04(s, 4H), 2.79 (t, J = 11.9 Hz, 1H), 1.50 (d, J = 6.7 Hz, 3H), 1.29 (dd, J =18.5, 4.7 Hz, 6H).

[0732] MS m / z (ES-API+): 528.20 [M+H]+

[0733] Example 68: Synthesis of 8-(4-chloro-2-fluorophenyl)-6-(2-(1-cyclobutyl-1H-pyrazol-4-yl)-6-methylmorpholinyl)-1,3-dimethylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 68)

[0734]

[0735] Dissolve 65-2 (100 mg, 0.21 mmol) in 5 mL of DMF. Add cesium carbonate (135 mg, 0.41 mmol) and bromocyclobutane (49 mg, 0.32 mmol), and react at 90 °C for 16 hours in the dark. Quench the crude reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 300%–50% PE / EA solution to obtain compound 68 (65 mg).

[0736] 1H NMR (300 MHz, CDCl3) δ 7.63 – 7.44 (m, 3H), 7.32 (ddt, J = 8.3,1.8, 0.9 Hz, 1H), 7.23 (ddd, J = 9.6, 3.7, 1.8 Hz, 1H), 4.83 – 4.62 (m, 2H), 4.57 (dd, J = 10.9, 2.7 Hz, 1H), 4.30 (t, J = 6.7 Hz, 1H), 3.84 – 3.67 (m,1H), 3.50 (s, 3H), 3.04 (s, 4H), 2.78 (dd, J = 13.2, 10.6 Hz, 1H), 2.62 –2.34 (m, 4H), 1.82 (s, 2H), 1.31 (d, J = 6.2 Hz, 2H), 1.27 – 1.23 (m, 1H).

[0737] MS m / z (ES-API+): 540.20 [M+H]+

[0738] Example 69: Synthesis of 8-(4-chloro-2-fluorophenyl)-6-(2-(1-(2-methoxyethyl)-1H-pyrazol-4-yl)-6-methylmorpholinyl)-1,3-dimethylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 69)

[0739]

[0740] Dissolve 65-2 (100 mg, 0.21 mmol) in 5 mL of DMF. Add cesium carbonate (135 mg, 0.41 mmol) and 1-iodo-2-methoxyethane (77 mg, 0.42 mmol), and react at 100 °C for 5 hours in the dark. Quench the crude reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 300%–20% PE / EA solution to obtain compound 69 (80 mg).

[0741] 1H NMR (300 MHz, CDCl3) δ 7.66 – 7.41 (m, 3H), 7.32 (dt, J = 8.3, 2.5Hz, 1H), 7.23 (d, J = 9.0 Hz, 1H), 4.73 (dd, J = 28.5, 13.4 Hz, 1H), 4.57 (d,J = 10.3 Hz, 1H), 4.26 (t, J = 5.3 Hz, 2H), 4.19 (s, 1H), 3.74 (t, J = 5.2Hz, 2H), 3.69 (d, J = 5.6 Hz, 1H), 3.51 (d, J = 2.0 Hz, 3H), 3.34 (d, J = 2.0Hz, 2H), 3.26 (s, 1H), 3.04 (d, J = 2.0 Hz, 4H), 2.78 (t, J = 12.0 Hz, 1H), 1.31 (d, J = 6.1 Hz, 3H)

[0742] MS m / z (ES-API+): 544.20 [M+H]+

[0743] Example 70: Synthesis of 8-(4-chloro-2-fluorophenyl)-6-(2-(1-(3-fluorocyclobut-2-en-1-yl)-1H-pyrazol-4-yl)-6-methylmorpholinyl)-1,3-dimethylpyrimidino[5,4-d]pyrimidin-2,4(1H,3H)-dione (compound 70)

[0744]

[0745] Dissolve 65-2 (100 mg, 0.21 mmol) in 5 mL of DMF. Add cesium carbonate (135 mg, 0.41 mmol) and 3-bromo-1,1-difluorocyclobutane (53 mg, 0.32 mmol), and react at 90 °C for 5 hours in the dark. Quench the crude reaction with water, extract three times with ethyl acetate, dry the organic layer with anhydrous MgSO4, filter and concentrate. Purify on a silica gel column, eluting with 300%–20% PE / EA solution to obtain compound 70 (50 mg).

[0746] 1H NMR (300 MHz, CDCl3) δ 7.70 (s, 1H), 7.63 (s, 1H), 7.59 (t, J = 7.6Hz, 1H), 7.39 – 7.29 (m, 1H), 7.25 – 7.18 (m, 1H), 6.57 (dd, J = 25.9, 14.0Hz, 1H), 4.89 – 4.55 (m, 4H), 4.52 – 4.34 (m, 1H), 3.81 (s, 1H), 3.57 – 3.44(m, 4H), 3.05 (d, J = 1.5 Hz, 3H), 2.97 (d, J = 12.2 Hz, 1H), 2.79 (t, J =11.9 Hz, 1H), 1.40 – 1.28 (m, 2H), 1.23 (d, J = 6.7 Hz, 1H).

[0747] MS m / z (ES-API+): 556.20 [M+H]+

[0748] Biological activity and related property test examples

[0749] Test Example 1: Detection of phosphorylation (pSyk) of human spleen tyrosine kinase in downstream cells:

[0750] Experimental principle:

[0751] The transmembrane glycoprotein TREM2 is activated upon binding to its extracellular ligand, thereby interacting with intracellular DAP12 and mediating the phosphorylation of the spleen tyrosine kinase Syk. Therefore, this study monitored TREM2 signaling via DAP12 by measuring Syk phosphorylation levels in the HEK-293T cell line overexpressing hTREM2 / DAP12.

[0752] Experimental materials:

[0753] Material Item number brand Trypsin-EDTA (0.25%), phenol red 25200056 Gibco 384 opti-plate 6007290 PerkinElmer 96-well plate 3599 corning DMEM 11995073 Thermofisher Fetal Bovine Serum (FBS) 10099141 Gibco Puromycin A11138-03 Gibco AlphaLISA SureFire Ultra p-SYK (Tyr525 / 526) Assay Kit ALSU-PSYK-A500 / 500 Assay Points PerkinElmer

[0754] Experimental methods:

[0755] HEK-293T cells overexpressing hTREM2 / DAP12 were routinely cultured in DMEM (containing 10% FBS and 7 μg / mL Luromycin). Cells were digested with Trypsin-EDTA, centrifuged, the supernatant was discarded, and the cells were resuspended in culture medium. The overexpressing cell density was adjusted to 50,000 cells / mL, and 100 μL was aliquoted into 96-well plates and incubated overnight at 37°C in a 5% CO2 incubator.

[0756] Aspirate the supernatant and add serum-free DMEM (containing 7 μg / mL Puromycin) medium. Starve the cells at 37°C and 5% CO2 for 2–3 h. Add the test compounds (concentrations of 30 pM, 1 nM, 30 nM, 1 μM, and 30 μM) prepared using serum-free DMEM (containing 7 μg / mL Puromycin) medium, and incubate at 37°C and 5% CO2 for 15 min. Remove the supernatant and prepare and add the corresponding reagents according to the AlphaLISA SureFire Ultra p-SYK kit instructions: 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 lysis buffer to a new 384-well plate, add 5 μL of acceptor mix to each well, vortex to mix, and incubate in the dark for 1 h. Add 5 μL of donor mix to each well, vortex to mix, and incubate overnight in the dark. Centrifuge at 1,000 rpm for 1 min, and use an αLISA program (E) on a microplate reader (brand: SpectraMax). x 680 nm, E m Read the readings for each well at 570 nm.

[0757] The test was performed in triplicate. The readings from the control group (containing no compound) were used as baseline 1 to calculate the relative readings (pSyk signal window, relative to medium only) for each concentration of the test compound. The relative readings were plotted against concentration, and the EC50 of the test compound was calculated using GraphPad Prism software with a four-parameter nonlinear fitting. 50 .

[0758] Test results:

[0759] “A” indicates EC 50 ≤50nM, "B" indicates 50nM < EC 50 ≤500nM, where “C” indicates 500nM < EC 50 ≤3000nM, “D” indicates EC 50 >3000nM.

[0760] Table 1: Effects of tested compounds on SYK phosphorylation in hTREM2 / DAP12-overexpressing HEK-293T cells

[0761] compound <![CDATA[EC 50 ]]> 1 B 2 B 3 A 5 B 6 B 7 A 8 C 9 B 10 C 12 A 13 A 14 B 15 B 16 B 17 B 18 B 19 B 20 B 21 B 22 A 23 A 24 A 25 A 26 A 27 A 28 B 30 A 31 A 32 A 33 A 34 A 35 B 36 A 37 B 38 A 39 A 40 A 41-P1 A 41-P2 B 42 A 43 A 44 B 45 D 46 D 47 D 48 B 49 A 50 A 51 A 52 A 53 B 54 A 55 A 56 A 57 A 58 B 59 B 60 B 61 A 62 A 63 A 64 A 65 A 66 A 67 A 68 B 69 B 70 B

Claims

1. A compound of formula (I) or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, （I） in, X1 is selected from CR 5 Or N; X2 is selected from CR 6 Or N; R 1 Selected from phenyl or 5-6-membered heteroaryl, wherein the phenyl or 5-6-membered heteroaryl group is optionally R 1a replace; R 1a Selected from halogens, OH, NH2, CN, C1-C6 alkyl, C3-C6 cycloalkyl, or 4-7 membered heterocyclic groups, 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 Selected from halogens, OH, NH2, CN, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy or C1-C6 haloalkoxy; R 3 and R 4 Independently selected from H, C1-C6 alkyl, C3-C6 cycloalkyl, or 4-7 membered heterocyclic groups, wherein the C1-C6 alkyl, C3-C6 cycloalkyl, or 4-7 membered heterocyclic group is optionally replaced by R. 3a replace; R 5 and R 6 Independently selected from H, halogen, OH, NH2, CN, 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. 5a replace; R 3a and R 5a It is independently selected from D, halogen, OH, NH2, CN, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy or C1-C6 haloalkoxy; R 2 Selected from or ; Y1 is selected from CR 8 Or N; Y2 is selected from C(R) 9 (R) 10 ) or C(O); Y3 is selected from C(R) 11 (R) 12 ); Y4 is selected from C(R) 13 (R) 14 ), NR 15 O or S; Y5 is selected from CR 16 Or N; Y6 is selected from C(R) 17 (R) 18 ); Y7 is selected from C(R) 19 2. NR 20 O, S, or S=O; Y9 is selected from key, C(R) 21 )2 or C=O; Y 13 Selected from bond, O or C(R) 22 )2; Y8, Y 10 Y 11 Y 12 Selected from C(R) 23 )2; Y 14 Selected from CR 24 Or N; m and n are independently selected from 0, 1, or 2; R 7 Selected from C1-C6 alkyl, C1-C6 alkylamino, C1-C6 alkoxy, C1-C6 haloalkyl, -C(O)O-(C1-C6 alkyl), C3-C6 cycloalkyl, 4-7 membered heterocyclic, phenyl, -O-phenyl, 5-6 membered heteroaryl, or -O-(5-6 membered heteroaryl), wherein the C1-C6 alkyl, C1-C6 alkylamino, C1-C6 alkoxy, C1-C6 haloalkyl, -C(O)O-(C1-C6 alkyl), C3-C6 cycloalkyl, 4-7 membered heterocyclic, phenyl, -O-phenyl, 5-6 membered heteroaryl, or -O-(5-6 membered heteroaryl) is optionally R 7a replace; R 7a Selected from O, halogen, CN, C1-C6 alkyl, C2-C6 alkenyl, C1-C6 alkoxy, C1-C6 alkylamino, C1-C6 haloalkyl, C3-C6 cycloalkyl, or 4-7 membered heterocyclic groups, wherein the C1-C6 alkyl, C2-C6 alkenyl, C1-C6 alkoxy, C1-C6 alkylamino, C1-C6 haloalkyl, C3-C6 cycloalkyl, or 4-7 membered heterocyclic group is optionally R 7b replace; R 7b Selected from OH, halogen, C1-C6 alkyl, C1-C6 alkoxy or -C(O)O-(C1-C6 alkyl); R 8 R 9 R 10 R 11 R 12 R 13 R 14 R 15 R 16 R 17 R 18 R 19 R 20 R 21 R 22 R 23 and R 24 Independently selected from H, D, halogen, OH, NH2, CN, C1-C6 alkyl, C3-C6 cycloalkyl, or 4-7 membered heterocyclic groups, wherein the OH, NH2, C1-C6 alkyl, C3-C6 cycloalkyl, or 4-7 membered heterocyclic group is optionally replaced by R. 8a Replace; or, R 9 and R 10 R 11 and R 12 R 13 and R 14 Or R 17 and R 18 The atoms bonded to it together form a C3-C6 cycloalkyl or a 4-7 membered heterocyclic group, wherein the C3-C6 cycloalkyl or 4-7 membered heterocyclic group is optionally R 8b replace; R 8a R 8b It is independently selected from D, halogen, OH, NH2, CN, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy or C1-C6 haloalkoxy.

2. The compound of formula (I) according to claim 1, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, X1 is selected from CH or N, or X1 is selected from N.

3. The compound of formula (I) according to claim 1 or 2, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, X2 is selected from CH or N.

4. The compound of formula (I) according to any one of claims 1 to 3, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R 1 Selected from phenyl, said phenyl optionally being R 1a replace.

5. The compound of formula (I) according to any one of claims 1 to 4, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R 1a Selected from halogens or C1-C6 alkyl groups, wherein the C1-C6 alkyl group is optionally R 1b replace.

6. The compound of formula (I) according to any one of claims 1 to 5, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R 1b Selected from halogens.

7. The compound of formula (I) according to any one of claims 1 to 6, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R 1 Selected from , or .

8. The compound of formula (I) according to any one of claims 1 to 7, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R 3 Selected from H or C1-C6 alkyl groups, wherein the C1-C6 alkyl group is optionally converted to R 3a replace.

9. The compound of formula (I) according to any one of claims 1 to 8, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R 4 Selected from H, C1-C6 alkyl or C3-C6 cycloalkyl, wherein the C1-C6 alkyl or C3-C6 cycloalkyl is optionally replaced by R. 3a replace.

10. The compound of formula (I) according to any one of claims 1 to 9, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R 2 Selected from , or , or R 2 Selected from , , , or , or R 2 Selected from , , , , , , , or .

11. The compound of formula (I) according to any one of claims 1 to 10, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R 7 Selected from phenyl or 5-6-membered heteroaryl, wherein the phenyl or 5-6-membered heteroaryl group is optionally R 7a Replace, or R 7 The group is selected from pyrazolyl, pyridyl, or pyrimidinyl, wherein the pyrazolyl, pyridyl, or pyrimidinyl group is optionally replaced by R. 7a Replace, or R 7 Selected from , or The , or Optionally R 7a Replace, or R 7 Selected from , , , , , , , , , , or .

12. The compound of formula (I) according to any one of claims 1 to 11, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, Compound (I) is selected from compound (II) or its stereoisomers or pharmaceutically acceptable salts. (II) Among them, R 1 R 2 R 3 and R 4 As defined in claim 1.

13. The compound of formula (I) according to any one of claims 1 to 11, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, Compound (I) is selected from compound (III) or its stereoisomers or pharmaceutically acceptable salts. (III) Among them, R 1 R 2 R 3 and R 4 As defined in claim 1.

14. The compound of formula (I) according to any one of claims 1 to 11, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, The compounds of formula (I) are selected from the following compounds or their stereoisomers or pharmaceutically acceptable salts, or .

15. A pharmaceutical composition, wherein, The pharmaceutical composition comprises a compound of formula (I) according to any one of claims 1 to 14, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

16. A method for treating a disease or condition related to loss of TREM2 function in mammals, comprising administering to the mammal in need, preferably a human, a therapeutically effective amount of the compound of formula (I) of any one of claims 1 to 14, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 15.

17. Use of the compound of formula (I) of any one of claims 1-14, or a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of claim 15, in the preparation of a medicament for the treatment or prevention of diseases or conditions associated with loss of TREM2 function in mammals, preferably humans.

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