TW202519219A - Preparation and application of aromatic KRAS mutant protein inhibitors - Google Patents

Abstract

The compound of the present invention has good activity in inhibiting tumor growth and good safety. The present invention discloses the preparation and application of Kras inhibitors. Specifically, the present invention provides a compound of formula (I) and a compound represented by formula (II) and a pharmaceutically acceptable salt thereof, a pharmaceutical composition containing the compound or its pharmaceutically acceptable salt, and a drug for treating or preventing Kras kinase-related diseases, especially tumors, using the compound or its pharmaceutically acceptable salt. This is a class of heterocyclic compounds. At the same time, the preparation method of the pharmaceutical composition of the compound or its pharmaceutically acceptable salt is disclosed. The substituents of the general formula (I) and formula (II) are the same as those defined in the specification.

Description

Preparation and application of aromatic KRAS mutant protein inhibitors

The present invention belongs to the field of drug synthesis, and specifically relates to a novel KRAS inhibitor and a preparation method and use thereof.

The present invention generally relates to novel compounds and methods for their preparation and use as KRAS inhibitors, e.g., for the treatment of cancer.

RAS represents a group of tightly associated monomeric globular proteins of 189 amino acids (molecular weight 21 kDa) that are associated with the plasma membrane and bind either GDP or GTP. RAS acts as a molecular switch. When RAS contains bound GDP, it is in a quiescent or closed state and is in an "inactive state". In response to exposure of the cell to certain growth-promoting stimuli, RAS is induced to convert its bound GDP to GTP. After binding to GTP, RAS is "switched on" and is able to interact with and activate other proteins (its "downstream targets"). The RAS proteins themselves have a very low intrinsic ability to hydrolyze GTP back to GDP, thus placing themselves in a closed state. Turning RAS off requires extrinsic proteins called GTPase activating proteins (GAPs), which interact with RAS and greatly speed up the conversion of GTP to GDP. Any mutation in RAS that affects its ability to interact with GAP or convert GTP back to GDP will result in a prolonged activation of the protein, leading to prolonged cell signaling that allows it to continue growing and dividing. Because these signals cause cells to grow and divide, overactive RAS signaling may ultimately lead to cancer.

Structurally, the RAS protein contains a G domain, which is responsible for the enzymatic activity of RAS - nucleotide binding and hydrolysis (GTPase reaction). It also contains a C-terminal extension called the CAAX box, which allows post-translational modification and is responsible for targeting the protein to the membrane. The size of the G domain is about 21-25 kDa, and it contains a phosphate binding ring (P-ring). The P-ring is the pocket where nucleotides bind in the protein. This is a rigid part of the domain with conserved amino acid residues (glycine 12, threonine 26 and lysine 16), which is essential for nucleotide binding and hydrolysis. The G domain also contains the so-called Switch I (residues 30-40) and Switch II (residues 60-76) regions, both of which are dynamic parts of the protein and are often referred to as a "spring-loaded" mechanism due to their ability to switch between the resting and loaded states. The key interaction is the hydrogen bond formed by Threonine 35 and Glycine 60, with the γ-phosphate of GTP, which holds the Switch1 and Switch2 regions, respectively, in their active conformations. After GTP hydrolysis and release of the phosphate, the two relax to the inactive GDP conformation.

The most well-known members of the RAS subfamily are HRAS, KRAS, and NRAS, mainly because they are associated with multiple types of cancer. Mutations in any of the three major isoforms of RAS (HRAS, NRAS, or KRAS) genes are the most common in human tumorigenesis. Approximately 30% of human tumors are found to carry mutations in the RAS gene. Of note, KRAS mutations are detected in 25-30% of tumors. In contrast, the incidence of oncogenic mutations in NRAS and HRAS family members is much lower (8% and 3%, respectively). The most common KRAS mutations are found at residues G12 and G13 of the P-loop and at residue Q61. G12C is a frequent mutation of the KRAS gene (glycine 12 mutates to cysteine). This mutation has been found in approximately 13% of cancer occurrences, approximately 43% of lung cancer occurrences, and approximately 100% of MYH-associated polyposis (familial colorectal cancer syndrome).

As a cutting-edge target, KRAS mutant protein has received widespread attention. Among them, AMG-510 developed by Amgen was approved for marketing by the FDA last year. In recent years, other companies have applied for a number of patents on KRAS inhibitors, such as W02016164675, W02016168540, WO2021141628, WO2022098625, WO2022087371, WO2020101736, WO2022109485, WO2022109487 and WO2020146613. Therefore, despite the progress made in this field, there is still a need for improved compounds and methods for treating cancer in this field, such as treating cancer by inhibiting KRAS, HRAS or NRAS. The present invention satisfies this need and provides other related advantages.

In short, the present invention provides compounds capable of inhibiting KRAS mutation, including stereoisomers, pharmaceutically acceptable salts, tautomers and prodrugs thereof.

A compound represented by general formula (I), its stereoisomers, pharmaceutically acceptable salts, or isomers, wherein the compound represented by general formula (I) has the following structure: (I) each L ₂ is independently selected at each occurrence from a bond, OC _0-6 alkyl, NHC _0-6 alkyl, C _1-6 alkyl, COC _0-6 alkyl or SC _0-6 alkyl; each R ₁ is independently selected at each occurrence from H, D, halogen, C _1-6 alkyl, -C _2-6 alkenyl, -C _2-6 alkynyl, CN, OC _1-6 alkyl; each R ₁ is independently optionally substituted or unsubstituted with 1, 2, 3, 4, 5 or 6 substituents selected from deuterium, halogen, -C _1-6 alkyl; n is 1-3; each X ₁ , X ₂ , X ₃ is independently selected at each occurrence from N, CR ₃ ; each R ₃ is independently selected from H, D, cyano, halogen, C _1-6 alkyl, CN; each R _{R 4} is independently selected from H, D, C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, CN, C _3-6 carbocyclyl, 3-10 membered heterocyclic ring, 4-10 membered heterofused ring; 3-8 membered heterocyclic ring contains 1, 2, 3 or 4 heteroatoms selected from N, O or S at each occurrence; each R ₄ is independently optionally substituted with 1, 2, 3, 4, 5 or 6 substituents selected from deuterium, halogen, C _1-6 alkyl, -C _1-6 alkoxy, oxo, OC _1-6 alkyl, C _3-6 carbocyclyl, 3-10 membered heterocyclic ring or unsubstituted; U is selected from 3-8 membered cycloalkyl, 3-8 membered heterocycloalkyl, 5-12 membered fused alkyl, 5-12 membered fused heterocyclo, 5-12 membered spirocyclo, 5-12 membered spiroheterocyclo, aromatic or heteroaromatic, each heterocycloalkyl, fused heterocyclo, spiroheterocyclo and heteroaromatic independently containing 1, 2, 3 or 4 heteroatoms selected from N, O, S or P at each occurrence, wherein the cycloalkyl, heterocycloalkyl, spirocyclo, fused cyclo, fused heterocyclo, spiroheterocyclo, aromatic or heteroaromatic is optionally substituted by one or more ^G1 ; ^G1 is independently selected from deuterium, cyano, halogen, _C1-6 alkyl, _C2-6 alkenyl, C _-OR5 , -OC(O)NR5R6, -C(O ₎ OR5, -C(O) _NR5R6 , -C(O _)R5 , -NR5R6, _-NR5C (O) _R6 , _-NR5C (O _{) NR6R7} , _-S ( _O ) _iR5 , _{or -NR5S} (O) _iR6 , wherein _the alkyl, _alkenyl , _alkynyl , _{cycloalkyl ,} heterocycloalkyl, _{aryl ,} heteroaryl is optionally substituted with one or more deuterium, cyano, halogen, _C1-7 alkyl, _C2-7 alkenyl, _C2-7 alkynyl, C R5, R6, R7, R8 _, R9 and R10 are _each independently _selected from hydrogen _, deuterium _, cyano, halogen, _C1-6 alkyl, _C2-6 alkyl, C3-7 alkyl, _C4-7 alkyl, C5-8 alkyl, _C6-7 alkyl, _{C7-10 aryl} , C8-12 aryl _{, C9-10} aryl, C1-6 alkyl, C1-6 alkyl, _{C2-7 alkyl} , C3-7 _alkyl , _C4-7 alkyl, _C5-8 alkyl, _C6-7 alkyl, C7-10 aryl, C8-12 aryl, _C9-10 aryl, _C8-12 alkyl, _C1-6 alkyl _{, C2-7} alkyl, _C3-7 alkyl, C4-7 alkyl, C5-8 alkyl, _C6-7 alkyl, C7-10 _aryl , C8-12 _aryl , _3-8 membered cycloalkyl or 3-8 membered monocyclic heterocyclic group, monocyclic heteroaryl group or phenyl group; and i is 1 or 2.

A compound represented by general formula (II), its stereoisomers, pharmaceutically acceptable salts, or isomers, wherein the compound represented by general formula (I) has the following structure: (II) each L ₂ is independently selected at each occurrence from a bond, OC _0-6 alkyl, NHC _0-6 alkyl, C _1-6 alkyl, COC _0-6 alkyl or SC _0-6 alkyl; each R ₁ is independently selected at each occurrence from H, D, halogen, C _1-6 alkyl, -C _2-6 alkenyl, -C _2-6 alkynyl, CN, OC _1-6 alkyl; each R ₁ is independently optionally substituted with 1, 2, 3, 4, 5 or 6 substituents selected from deuterium, halogen, -C _1-6 alkyl or unsubstituted; n is 1-3; each X ₁ is independently selected at each occurrence from N, CR ₃ ; each R ₃ is independently selected from H, D, cyano, halogen, C _1-6 alkyl, CN; Y ₁ , Y ₂ , Y ₃ , Y ₄ at most one selected from the heteroatom N, O, S, the others selected from CR ₃ ; each R ₃ is independently selected from H, D, cyano, halogen, C _1-6 alkyl, CN; each R ₄ is independently selected from H, D, C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, CN, C _3-6 carbocyclic group, 3-10 membered heterocyclic ring, 4-10 membered heterofused ring; 3-8 membered heterocyclic ring contains 1, 2, 3 or 4 heteroatoms selected from N, O, or S at each occurrence; each R ₄ is independently optionally substituted by 1, 2, 3, 4, 5 or 6 deuterium, halogen, C _1-6 alkyl, -C _1-6 alkoxy, oxo, OC _1-6 alkyl, C G is selected from _3-8 membered cycloalkyl, 3-8 membered heterocycloalkyl, 5-12 membered fused alkyl, 5-12 membered fused heterocyclo, 5-12 membered spirocyclo, 5-12 membered spiroheterocyclo, aromatic or heteroaromatic, each heterocycloalkyl, fused heterocyclo, spiroheterocyclo and heteroaromatic independently containing 1, 2, 3 or 4 heteroatoms selected from N, O, S or P at each occurrence, wherein the cycloalkyl, heterocycloalkyl, spirocyclo, fused heterocyclo, fused heterocyclo, spiroheterocyclo, aromatic or heteroaromatic is optionally substituted by one or more G1; G is selected from 3-8 membered cycloalkyl, 3-8 membered heterocycloalkyl, 5-12 membered fused heterocyclo, 5-12 membered spirocyclo, 5-12 membered spiroheterocyclo, aromatic or heteroaromatic, each heterocycloalkyl, fused heterocyclo, spiroheterocyclo and heteroaromatic independently containing 1, 2, 3 or 4 heteroatoms selected from N, O, S or P at each occurrence, wherein the cycloalkyl, heterocycloalkyl, spirocyclo, fused heterocyclo, fused heterocyclo, spiroheterocyclo, aromatic or heteroaromatic is optionally substituted by one or more ^G1 ; ¹ are each independently selected from deuterium, cyano, halogen, C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-8 cycloalkyl or 3-8 membered heterocyclic group, C _6-10 aryl, 5-10 membered heteroaryl, -OR ₅ , -OC(O)NR ₅ R ₆ , -C(O)OR ₅ , -C(O)NR ₅ R ₆ , -C(O)R ₅ , -NR ₅ R ₆ , -NR ₅ C(O)R ₆ , -NR ₅ C(O)NR ₆ R ₇ , -S(O) _i R ₅ or -NR ₅ S(O) _i R ₆ wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl is optionally substituted with one or more deuterium, cyano, halogen, _C1-7 alkyl, _C2-7 alkenyl, _C2-7 alkynyl, _C3-9 cycloalkyl or 3-9 membered heterocyclo, _C7-10 aryl, 6-10 membered heteroaryl, _-OR8 , -OC(O) _NR8R9 , -C(O) _OR8 , _-C (O) _NR8R9 , -C(O) _R8 , _-NR8R9 , _-NR8C (O _{)R9 , -NR8C} (O) _{NR9R10 , -S} (O _{)iR8 or -NR8S} (O) _iR9 substituents _{; R5} , _R6 R ₇ , R ₈ , R ₉ and R ₁₀ are each independently selected from hydrogen, deuterium, cyano, halogen, C _1-6 alkyl, C _3-8 cycloalkyl or 3-8 membered monocyclic heterocyclic group, monocyclic heteroaromatic group or phenyl; and i is 1 or 2.

In some embodiments, the compounds of formula (I) and formula (II) or their isomers, solvates or precursors, or their pharmaceutically acceptable salts are selected from the following compounds, their isomers, solvates or precursors, or their pharmaceutically acceptable salts: .

On the other hand, the present invention also provides a pharmaceutical composition comprising a compound represented by formula (I) and formula (II) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

On the other hand, the present invention relates to a method for treating a disease associated with KRas, KRas G12A, KRas G12C, KRas G12D, KRas G12R, KRas G12S, KRas G12V, KRas G13D or KRas Q61H in a mammal, comprising administering a therapeutically effective amount of a compound represented by formula (I) and formula (II) or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof to a mammal, preferably a human, in need of such treatment.

On the other hand, the present invention relates to the use of the compounds represented by formula (I) and formula (II) or pharmaceutically acceptable salts thereof in drugs for preventing or treating KRas, KRas G12A, KRas G12C, KRas G12D, KRas G12R, KRas G12S, KRas G12V, KRas G13D or KRas Q61H related diseases.

On the other hand, the present invention relates to compounds represented by formula (I) and formula (II) or their pharmaceutically acceptable salts, or pharmaceutical compositions thereof, for preventing or treating diseases related to KRas, KRas G12A, KRas G12C, KRas G12D, KRas G12R, KRas G12S, KRas G12V, KRas G13D or KRas Q61H.

Some chemical terms

Unless otherwise stated, the following terms are used in the specification and patent application.

The expression "C _xy " used in this article has the following meanings and represents the range of carbon number, wherein x and y are both integers, for example, C _3-8 cycloalkyl represents a cycloalkyl having 3-8 carbon atoms, i.e., a cycloalkyl having 3, 4, 5, 6, 7 or 8 carbon atoms. It should also be understood that "C _3-8 " also includes any sub-range therein, such as C _3-7 , C _3-6 , C _4-7 , C _4-6 , C _5-6 , etc.

"Alkyl" refers to a straight or branched alkyl group containing 1 to 20 carbon atoms, such as 1 to 18 carbon atoms, 1 to 12 carbon atoms, 1 to 8 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms. Non-limiting examples of alkyl include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2 dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl and 2-ethylbutyl. The alkyl may be substituted or unsubstituted.

"Alkenyl" refers to a straight or branched alkyl group containing at least one carbon-carbon double bond and typically 2 to 20 carbon atoms, such as 2 to 8 carbon atoms, 2 to 6 carbon atoms, or 2 to 4 carbon atoms. Non-limiting examples of alkenyl include vinyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 2-methyl-2-propenyl, 1,4-pentadienyl, and 1,4-butadienyl. The alkenyl group may be substituted or unsubstituted.

"Alkynyl" refers to a straight or branched alkyl group containing at least one carbon-carbon triple bond and typically 2 to 20 carbon atoms, such as 2 to 8 carbon atoms, 2 to 6 carbon atoms, or 2 to 4 carbon atoms. Non-limiting examples of alkynyl include ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, and 3-butynyl. The alkynyl may be substituted or unsubstituted.

"Cycloalkyl" refers to a saturated cyclic alkyl substituent containing 3 to 14 carbon ring atoms. A cycloalkyl can be a single carbon ring, typically containing 3 to 7 carbon ring atoms. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl. A cycloalkyl can alternatively be a di- or tri-ring fused together, such as decahydronaphthyl, and the cycloalkyl can be substituted or unsubstituted.

"Heterocyclic", "heterocycloalkyl", "heterocycle" refers to a stable 3-18 membered monovalent non-aromatic ring, including 2-12 carbon atoms, 1-6 heteroatoms selected from nitrogen, oxygen and sulfur. Unless otherwise specified, the heterocyclic group can be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may contain fused rings, spirocyclic or bridged ring systems, and the nitrogen, carbon or sulfur on the heterocyclic group can be optionally oxidized, the nitrogen atom can be optionally quaternized, and the heterocyclic group can be partially or fully saturated. The heterocyclic group can be connected to the rest of the molecule through a single bond through a carbon atom or heteroatom on the ring. The heterocyclic group containing fused rings may contain one or more aromatic rings or heteroaromatic rings, as long as the atoms on the non-aromatic rings are connected to the rest of the molecule. For the purpose of the present invention, the heterocyclic group is preferably a stable 4-11-membered monovalent non-aromatic monocyclic or bicyclic ring containing 1-3 heteroatoms selected from nitrogen, oxygen and sulfur, and more preferably a stable 4-8-membered monovalent non-aromatic monocyclic ring containing 1-3 heteroatoms selected from nitrogen, oxygen and sulfur. Non-limiting examples of heterocyclic groups include azacycloheptanyl, azacyclobutyl, decahydroisoquinolinyl, dihydrofuranyl, dihydroindolyl, dioxolanyl, 1,1-dihydro-thiofurinyl, imidazolidinyl, imidazolinyl, isothiazolidinyl, isoxazolidinyl, oxolinyl, octahydroindolyl, octahydroisoindolyl, oxazinyl, piperazinyl, piperidinyl, 4-piperidinonyl, pyranyl, pyrazolidinyl, pyrrolidinyl, quinolizinyl, quininyl, tetrahydrofuranyl, tetrahydropyranyl, and the like.

"Spiroheterocyclic group" refers to a polycyclic heterocyclic group of 5 to 20 members, in which one atom (called spiro atom) is shared between monocyclic rings, wherein one or more ring atoms are heteroatoms selected from nitrogen, oxygen or S(O) _m (wherein m is an integer from 0 to 2), and the remaining ring atoms are carbon. These may contain one or more double bonds, but none of the rings has a completely conjugated electronic system, preferably 6 to 14 members, more preferably 7 to 10 members. Spirocycloalkyl groups are divided into monospiroheterocyclic groups, bispiroheterocyclic groups or polyspiroheterocyclic groups according to the number of spiro atoms shared between rings, preferably monospirocycloalkyl groups and bispirocycloalkyl groups. More preferably, it is a 4-membered/4-membered, 4-membered/5-membered, 4-membered/6-membered, 5-membered/5-membered or 5-membered/6-membered monospirocyclic group. Non-limiting examples of spiroheterocyclic groups include:

"Fused heterocyclic group" refers to a polycyclic heterocyclic group of 5 to 20 members, each ring in the system shares a pair of adjacent atoms with other rings in the system, one or more rings may contain one or more double bonds, but no ring has a completely conjugated π electron system, wherein one or more ring atoms are heteroatoms selected from nitrogen, oxygen or S(O) _m (wherein m is an integer from 0 to 2), and the remaining ring atoms are carbon. Preferably, it is 6 to 14 members, and more preferably, it is 7 to 10 members. According to the number of constituent rings, it can be classified into bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic alkyl groups, preferably bicyclic or tricyclic, more preferably 5-membered/5-membered or 5-membered/6-membered bicyclic fused heterocyclic groups. Non-limiting examples of fused heterocyclic groups include:

"Aryl" or "aromatic group" refers to an aromatic monocyclic or fused polycyclic group containing 6 to 14 carbon atoms, preferably 6 to 10 members, such as phenyl and naphthyl, more preferably phenyl. The aryl ring can be fused to a heteroaryl, heterocyclic or cycloalkyl ring, wherein the ring connected to the parent structure is the aryl ring.

"Heteroaryl" or "heteroaromatic group" refers to a 5-16 membered ring system containing 1-15 carbon atoms, preferably 1-10 carbon atoms, 1-4 heteroatoms selected from nitrogen, oxygen and sulfur, and at least one aromatic ring. Unless otherwise specified, the heteroaryl group can be a monocyclic, bicyclic, tricyclic or tetracyclic ring system, which may contain fused or bridged ring systems, as long as the point of attachment to the rest of the molecule is an aromatic ring atom. The nitrogen atoms, carbon atoms and sulfur atoms on the heteroaromatic ring can be selectively oxidized, and the nitrogen atoms can be selectively quaternized. For the purpose of the present invention, the heteroaryl group is preferably a stable 4-11 membered monoaromatic ring containing 1-3 heteroatoms selected from nitrogen, oxygen and sulfur, and more preferably a stable 5-8 membered monoaromatic ring containing 1-3 heteroatoms selected from nitrogen, oxygen and sulfur. Non-limiting examples of heteroaryl groups include acridinyl, azazepinyl, benzimidazolyl, benzindolyl, benzodioxinyl, benzodioxolyl, benzofuranonyl, benzofuranyl, benzonaphthofuranyl, benzopyroneonyl, benzopyranyl, benzopyrazolyl, benzothiadiazolyl, benzothiazolyl, benzotriazolyl, furanyl, imidazolyl, indazolyl, indolyl, oxazolyl, purinyl, pyrazinyl, pyrazolyl, oxazinyl, pyridinyl, pyrimidinyl, pyrrolyl, quinazolinyl, quinolinyl, quininyl, tetrazolyl, thiadiazolyl, thiazolyl, thienyl, triazinyl, triazolyl, and the like. In the present invention, the heteroaryl group is preferably a 5-8 membered heteroaryl group containing 1-3 heteroatoms selected from nitrogen, oxygen and sulfur, more preferably a pyridyl group, a pyrimidyl group, a thiazolyl group. The heteroaryl group may be substituted or unsubstituted.

"Halogen" refers to fluorine, chlorine, bromine or iodine.

"Hydroxy" refers to -OH, "amino" refers to -NH ₂ , "amido" refers to -NHCO-, "cyano" refers to -CN, "nitro" refers to -CN, "isocyano" refers to -NC, and "trifluoromethyl" refers to -CF ₃ .

The term "impurity atom" or "impurity" as used herein alone or as part of other components refers to atoms other than carbon and hydrogen, and the impurity atoms are independently selected from oxygen, nitrogen, sulfur, phosphorus, silicon, selenium and tin, but are not limited to these atoms. In embodiments where two or more impurity atoms are present, the two or more impurity atoms may be the same as each other, or some or all of the two or more impurity atoms may be different.

The terms "fused" or "fused ring" as used herein, alone or in combination, refer to a cyclic structure in which two or more rings share one or more bonds.

[0046] The term "spiro" or "spirocyclic," as used herein, alone or in combination, refers to a ring structure in which two or more rings share one or more atoms.

"Optional" or "optionally" means that the subsequently described event or circumstances may but need not occur, and that the description includes instances where the event or circumstances occur and where they do not occur. For example, "a heterocyclic group optionally substituted with an alkyl group" means that the alkyl group may but need not be present, and that the description includes instances where the heterocyclic group is substituted with an alkyl group and instances where the heterocyclic group is not substituted with an alkyl group.

"Substituted" means that one or more atoms, preferably 5, more preferably 1 to 3 atoms, in a group are independently substituted by a corresponding number of substituents. It goes without saying that the substituents are in their possible chemical positions and the skilled person can determine (by experiment or theory) possible or impossible substitutions without undue effort. For example, a free amine or hydroxyl group may be unstable when combined with a carbon atom with an unsaturated (such as olefin) bond. The substituents include but are not limited to hydroxyl, amine, halogen, cyano, C _1-6 alkyl, C _1-6 alkoxy, C _2-6 alkenyl, C _2-6 alkynyl, C _3-8 cycloalkyl, etc.

"Pharmaceutical composition" refers to a composition containing one or more compounds described herein or their pharmaceutically acceptable salts or prodrugs and other components such as pharmaceutically acceptable carriers and excipients. The purpose of the pharmaceutical composition is to facilitate administration to an organism, facilitate the absorption of the active ingredient, and thereby exert biological activity.

"Isomers" refer to compounds with the same molecular formula but different properties or orders of atomic bonding or spatial arrangements of their atoms, and isomers with different atomic spatial arrangements are called "stereoisomers". Stereoisomers include optical isomers, geometric isomers and conformational isomers. The compounds of the present invention may exist in the form of optical isomers. Depending on the configuration of substituents around the chiral carbon atom, these optical isomers are "R" or "S" configurations. Optical isomers include enantiomers and diastereomers, and methods for preparing and separating optical isomers are known in the art.

The compounds of the present invention may also exist in geometric isomers. The present invention contemplates various geometric isomers and mixtures thereof resulting from the distribution of substituents around carbon-carbon double bonds, carbon-nitrogen double bonds, cycloalkyl or heterocyclic groups. Substituents around carbon-carbon double bonds or carbon-nitrogen bonds are designated as Z or E configurations, and substituents around cycloalkyl or heterocyclic groups are designated as cis or trans configurations.

The compounds of the present invention may also exhibit tautomerism, such as keto-enol tautomerism.

It should be understood that the present invention includes any tautomeric or stereoisomeric forms and mixtures thereof, and is not limited to any one tautomeric or stereoisomeric form used in the naming or chemical formula of the compound.

"Isotopes" are all isotopes of atoms that occur in the compounds of the present invention. Isotopes include those atoms with the same atomic number but different mass numbers. Examples of isotopes suitable for incorporation into the compounds of the present invention are hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, such as but not limited to ^2H , ^3H , ^13C , ^14C , ^15N , ^18O , ^31P , ^32P , ^35S , ^18F and ^36Cl , respectively. Isotope-labeled compounds of the present invention can generally be prepared by conventional techniques known to those skilled in the art or by methods similar to those described in the attached Examples using appropriate isotope-labeled reagents in place of non-isotope-labeled reagents. Such compounds have a variety of potential uses, such as as standards and reagents in determining biological activity. In the case of stable isotopes, such compounds have the potential to favorably alter biological, pharmacological, or pharmacokinetic properties.

"Prodrug" means that the compounds of the present invention can be administered in the form of prodrugs. Prodrugs are derivatives that are converted into the biologically active compounds of the present invention under physiological conditions in vivo, such as by oxidation, reduction, hydrolysis, etc. (each of which is carried out with or without enzyme participation). Examples of prodrugs are compounds wherein an amine group in the compounds of the present invention is acylated, alkylated or phosphorylated, such as an eicosylamino group, a propylamino group, a neopentanoyloxymethylamino group, or wherein a hydroxyl group is acylated, alkylated, phosphorylated or converted to a borate, such as an acetyloxy group, a palmityloxy group, a neopentanoyloxy group, a succinyloxy group, a fumaroyloxy group, a propylamino group, or wherein a carboxyl group is esterified or amidated, or wherein a hydroxyl group forms a disulfide bridge with a carrier molecule, such as a peptide, which selectively delivers a drug to a target and/or to the cytosol of a cell. These compounds can be prepared from the compounds of the present invention according to known methods.

"Pharmaceutically acceptable salt" or "pharmaceutically acceptable" refers to a compound made from a pharmaceutically acceptable base or acid, including an inorganic base or acid and an organic base or acid. In the case where the compounds of the present invention contain one or more acidic or alkaline groups, the present invention also includes their corresponding pharmaceutically acceptable salts. Thus, the compounds of the present invention containing an acidic group may exist in the form of a salt and may be used according to the present invention, for example as an alkali metal salt, an alkaline earth metal salt or as an ammonium salt. More specific examples of such salts include sodium salts, potassium salts, calcium salts, magnesium salts or with amines or organic amines, such as primary amines, secondary amines, tertiary amines, cyclic amines, etc., such as ammonia, isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, diethanolamine, ethanolamine, dicyclohexylamine, ethylenediamine, purine, piperazine, piperidine, choline and caffeine. Particularly preferred organic bases are salts of isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline and caffeine. The compounds of the present invention containing basic groups can exist in the form of salts and can be used according to the present invention in the form of additions thereof with inorganic or organic acids. Examples of suitable acids include hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, phosphoric acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalene disulfonic acid, oxalic acid, acetic acid, tartaric acid, lactic acid, salicylic acid, benzoic acid, formic acid, propionic acid, pivalic acid, malonic acid, succinic acid, pimelic acid, fumaric acid, maleic acid, apple acid, aminosulfonic acid, phenylpropionic acid, gluconic acid, ascorbic acid, isonicotinic acid, citric acid, adipic acid and other acids known to those skilled in the art. If the compounds of the present invention contain both acidic and basic groups in the molecule, the present invention also includes internal salts or internal ammonium salts in addition to the salt forms mentioned. The respective salts are obtained by customary methods known to the person skilled in the art, for example by contacting these with organic or inorganic acids or bases in a solvent or dispersion or by anion exchange or cation exchange with other salts.

Therefore, when referring to "compound", "compound of the present invention" or "compound of the present invention" in the present invention, all forms of the compound, such as prodrugs, stable isotopic derivatives, pharmaceutically acceptable salts, isomers, meso- and racemates, enantiomers, diastereomers and mixtures thereof are included.

As used herein, the term "tumor" includes benign tumors and malignant tumors (eg, cancer).

In this article, the term "cancer" includes various malignant tumors in which KRAS is involved, including but not limited to pancreatic cancer, non-small cell lung cancer, small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, colon cancer, colorectal cancer, thyroid cancer, embryonal rhabdomyosarcoma, cutaneous granuloma, melanoma, liver cancer, rectal cancer, bladder cancer, throat cancer, breast cancer, Cancer, prostate cancer, neuroglioma, ovarian cancer, head and neck squamous cell cancer, cervical cancer, esophageal cancer, kidney cancer, skin cancer, lymphoma, gastric cancer, acute myeloid leukemia, myelofibrosis, B-cell lymphoma, monocytic leukemia, splenomegaly, eosinophilic syndrome, myeloma and other solid tumors and blood tumors.

As used herein, the term "effective amount", "therapeutically effective amount" or "pharmaceutically effective amount" refers to an amount of at least one agent or compound sufficient to relieve to some extent one or more symptoms of the disease or condition being treated after administration. The result can be the elimination and/or alleviation of signs, symptoms or causes of disease or any other desired change in a biological system. For example, an "effective amount" for treatment is the amount of a composition comprising a compound disclosed herein that is required to provide a significant symptom-relieving effect clinically. Techniques such as dose escalation trials can be used to determine the effective amount appropriate for any individual case.

The term "polymorph" or "polymorphic form (phenomenon)" used in the present invention means that the compounds of the present invention have multiple crystal lattice forms. Some compounds of the present invention may have more than one crystal form. The present invention covers all multiple forms or mixtures thereof.

The intermediate compounds of the compounds of the present invention and their polymorphs are also within the scope of the present invention.

Crystallization often produces a solvate of the compound of the present invention. The term "solvate" as used herein refers to a complex composed of one or more molecules of the compound of the present invention and one or more solvent molecules.

The solvent may be water, in which case the solvate is a hydrate. It may also be an organic solvent. Therefore, the compounds of the present invention may exist as hydrates, including monohydrates, dihydrates, hemihydrates, trihydrates, tetrahydrates, etc., and corresponding solvated forms. The compounds of the present invention may be true solvates, but in some other cases, the compounds of the present invention may also only accidentally retain water or a mixture of water and some other solvents. The compounds of the present invention may react in a solvent or precipitate or crystallize in a solvent. Solvates of the compounds of the present invention are also included in the scope of the present invention.

As used herein, the term "acceptable" with respect to a formulation, composition or ingredient means having no persistent detrimental effect on the general health of the treated subject.

The term "pharmaceutically acceptable" as used herein refers to a substance (such as a carrier or diluent) that does not affect the biological activity or properties of the compounds of the present invention and is relatively non-toxic, i.e., the substance can be administered to a subject without causing adverse biological reactions or interacting in an adverse manner with any components contained in the composition.

"Pharmaceutically acceptable carriers" include but are not limited to adjuvants, carriers, excipients, auxiliary agents, deodorants, diluents, preservatives, dyes/colorants, flavor enhancers, surfactants and wetting agents, dispersants, suspending agents, stabilizers and other penetrants, solvents, or emulsifiers that have been approved by relevant government administrative departments for use in humans and domesticated animals.

The terms "subject", "patient", "subject" or "individual" as used herein refer to individuals suffering from a disease, disorder or condition, etc., including mammals and non-mammals. Examples of mammals include, but are not limited to, any member of the class Mammalia: humans, non-human primates (e.g., chimpanzees and other apes and monkeys); livestock, such as cattle, horses, sheep, goats, pigs; domestic animals, such as rabbits, dogs and cats; laboratory animals, including rodents, such as rats, mice and guinea pigs, etc. Examples of non-human mammals include, but are not limited to, birds and fish, etc. In one embodiment of the methods and compositions provided herein, the mammal is a human.

The term "treatment" as used herein refers to the treatment of relevant diseases and conditions in mammals, especially humans, including: (i) preventing mammals, especially mammals that have been previously exposed to a disease or condition but have not yet been diagnosed with the disease or condition, from developing the corresponding disease or condition; (ii) inhibiting the disease or condition, that is, controlling its development; (iii) alleviating the disease or condition, that is, causing the disease or condition to regress and slow down; (iv) alleviating the symptoms caused by the disease or condition.

As used herein, the terms "disease" and "symptom" may be used interchangeably or may have different meanings because certain specific diseases or symptoms do not yet have a known causative agent (so the cause of the disease is still unclear) and therefore cannot be considered a disease but can only be considered an unwanted condition or syndrome, of which more or less specific symptoms have been confirmed by clinical researchers.

The terms "administering", "applying", "administering" and the like as used herein refer to methods that can deliver a compound or composition to a desired site for biological action. These methods include but are not limited to oral routes, transduodenal routes, parenteral injection (including intravenous, subcutaneous, intraperitoneal, intramuscular, intraarterial injection or infusion), local administration and rectal administration. In a preferred embodiment, the compounds and compositions discussed herein are administered orally.

The present invention also provides a method for preparing the compound. The preparation of the compound described in the general formula (I) of the present invention can be completed by the following exemplary methods and examples, but these methods and examples should not be considered as limiting the scope of the present invention in any way. The compound described in the present invention can also be synthesized by synthetic techniques known to those skilled in the art, or a combination of methods known in the art and the methods described in the present invention. The product obtained in each step is obtained by separation techniques known in the art, including but not limited to extraction, filtration, distillation, crystallization, chromatographic separation, etc. The starting materials and chemical reagents required for the synthesis can be conventionally synthesized or purchased according to the literature (reaxys).

Unless otherwise stated, temperatures are in degrees Celsius. Reagents were purchased from commercial suppliers such as Chem blocks Inc and 3wpharm and were used directly without further purification unless otherwise stated.

Unless otherwise stated, the following reactions were performed at room temperature, in anhydrous solvents, under a positive pressure of nitrogen or gas, or using a desiccator; glassware was oven-dried and/or heat-dried.

Unless otherwise stated, column chromatography purification used 200-300 mesh silica gel from Qingdao Ocean Chemical Plant; thin layer chromatography silica gel prefabricated plates (HSGF254) produced by Yantai Institute of Chemical Industry were used for thin layer chromatography; MS was determined using a Therno LCD Fleet (ESI) liquid chromatography-mass spectrometer.

The nuclear magnetic resonance data (1H NMR) were obtained using a Bruker Avance-400MHz or Varian Oxford-400Hz NMR instrument. The solvents used for the nuclear magnetic resonance data included CDCl3, CD3OD, D2O, DMS-d6, etc., with tetramethylsilane (0.000ppm) as the reference or residual solvent as the reference (CDCl3: 7.26ppm; CD3OD: 3.31ppm; D2O: 4.79ppm; d6-DMSO: 2.50ppm). When peak shape diversity is indicated, the following abbreviations are used to represent different peak shapes: s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet), br (broad peak), dd (double of doublets), dt (double of triplets). If coupling constants are given, they are in Hertz (Hz).

Preparation of intermediates

3- Methoxyisoquinoline -1- pinacol borate

Step A: To a solution of 3-methoxy-2H-isoquinolin-l-one (6.16 g, 20 mmol), DIEA (7.7 g, 60 mmol) in DCM (120 mL) was added Tf ₂ O (8.41 g, 30 mmol), and the mixture was stirred at -40 °C for 0.5 h. The reaction mixture was diluted with ice water (50 mL), and then extracted with DCM (30 mL). The combined organic phases were dried over Na ₂ SO ₄ and concentrated to dryness. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate = 1/0 to 60/1) to give 1-trifluoromethanesulfonyloxy-3-methoxyisoquinoline (5.89 g, 96% yield). LC-MS(ESI): m/z 308[M+H] ⁺ .

Step B: To a mixture of 1-trifluoromethanesulfonyloxy-3-methoxyisoquinoline (5.83 g, 19 mmol), 4,4,5,5-tetramethyl-2-(4,4,1,5,5-tetramethyl-1,3,2-dioxoborolan-2-yl)-1,2,2-dioxyborane (10 g, 20 mmol), AcOK (5.78 g, 60 mmol) in toluene (110 mL) was added Pd(dppf)Cl ₂ (1.44 g, 2 mmol). The mixture was degassed and stirred at 130 °C for 3 hours. The reaction mixture was filtered and concentrated to obtain a residue. To the residue was added EtOAc (100 mL) and water (80 mL). The organic phase was washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give a residue. The residue was purified by column chromatography (SiO2) and eluted with 30-40% ethyl acetate in petroleum ether to give 3-methoxyisoquinoline-1-pinacol borate (3.68 g, 68% yield). LC-MS (ESI): m/z 286 [M+H] ⁺ .

(2- Fluoro -6- methoxy -8-(4,4,5,5 -tetramethyl -1,3,2- dioxoboran -2- yl ) isoquinolin -1- yl ) ethynyl ) triisopropylsilane

Step A: To a stirred solution of 5-fluoro-2-methylbenzoic acid (38.4 g) in DMF (450 ml) were added NIS (61.7 g) and sodium (II) acetate (5.6 g) at room temperature. The resulting reaction mixture was heated at 110°C for 15 h. The reaction mixture was cooled to room temperature and poured into water (1000 ml). The resulting mixture was extracted with EtOAc (2 x 400 ml). The combined organic phases were washed with brine (300 _ml ), dried over _Na2SO4 , filtered and concentrated under reduced pressure to give 3-fluoro-2-iodo-6-methylbenzoic acid (49.6 g). The material was used directly in the next step without further purification.

Step B: Heat a solution of 3-fluoro-2-iodo-6-methylbenzoic acid (42 g, 0.15 mol), _SOCl2 (120 mL) and DMF (6 mL) until the mixture becomes homogeneous (15 min). Keep the solution at 23 °C for another 30 min and then concentrate the solution. Add MeOH (240 mL) to the crude residue and keep the solution at 23 °C for 30 min. Concentrate the solution to give methyl 3-fluoro-2-iodo-6-methylbenzoate (43 g). This material was used directly in the next step without further purification.

Step C: To a solution of methyl 3-fluoro-2-iodo-6-methylbenzoate (22 g, 75 mmol) in CCl ₄ (130 mL) was added N-bromosuccinimide (22 g, 123 mmol) and benzoyl peroxide (1.46 g, 7.8 mmol). The reaction mixture was stirred at 85°C under nitrogen atmosphere for 3 hours. The crude product was filtered and washed with toluene. The volatiles were evaporated, the residue was dissolved in DMF (150 mL), and sodium cyanide (7.50 g) was added. The resulting mixture was heated at 60°C for 5 hours. The reaction mixture was cooled to room temperature, treated with water (50 mL), and extracted with ethyl acetate (3 x 50 mL). The combined organics were washed with saline solution (50 mL) and dried over anhydrous sodium sulfate. The solution was filtered and concentrated under reduced pressure to give the crude product. Elution with 30-40% ethyl acetate in petroleum ether gave methyl 3-fluoro-2-iodo-6-(cyanomethyl)benzoate (14.2 g). LC-MS (ESI): m/z 320 [M+H] ⁺ .

Step D: To a solution of methyl 3-fluoro-2-iodo-6-(cyanomethyl)benzoate (10.24 g, 32 mmol) in methanol (95 mL) was added sodium methoxide (25% in methanol, 34.6 g, 35.7 mL). The resulting mixture was heated at 70 °C for 4 h. The reaction mixture was cooled to room temperature and acidified using 1N HCl. The precipitated solid was filtered, washed with water and dried under vacuum to give 7-fluoro-8-iodo-3-methoxy-2H-isoquinolin-l-one (10.55 g, 103% yield) as a yellow solid. LC-MS (ESI): m/z 320 [M+H] ⁺ .

Step E: Triisopropylsilyl acetylene (5.47 g, 0.03 mol) and triethylamine (14.4 ml), then copper iodide (0.2 g) and bis-(triphenylphosphine)-palladium dichloride (0.73 g) were added to an oxygen-free solution of 7-fluoro-8-iodo-3-methoxy-2H-isoquinolin-l-one (9.6 g, 0.03 mol) in 150 ml of dry THF under argon. The solution was stirred at ambient temperature for 16 hours, then filtered through celite and evaporated. The residue was purified by flash chromatography (SiO ₂ ) eluting with 30-40% ethyl acetate in petroleum ether to give 7-fluoro-8-((triisopropylsilyl)ethynyl)-3-methoxy-2H-isoquinolin-1-one (8.7 g, 78% yield). LC-MS (ESI): m/z 374 [M+H] ⁺ .

Step F: To a solution of 7-fluoro-8-((triisopropylsilyl)ethynyl)-3-methoxy-2H-isoquinolin-l-one (7.4 g, 20 mmol), DIEA (7.7 g, 60 mmol) in DCM (120 mL) was added Tf ₂ O (8.41 g, 30 mmol) and the mixture was stirred at -40°C for 0.5 h. The reaction mixture was diluted with ice water (50 mL) and then extracted with DCM (30 mL). The combined organic phases were dried over Na ₂ SO ₄ and concentrated to dryness. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate = 1/0 to 60/1) to give 1-trifluoromethanesulfonyl-7-fluoro-3-methoxy-8-((triisopropylsilyl)ethynyl)isoquinoline (9.9 g, 98% yield). LC-MS (ESI): m/z 506 [M+H] ⁺ .

Step G: To a mixture of 1-trifluoromethanesulfonyl-7-fluoro-3-methoxy-8-((triisopropylsilyl)ethynyl)isoquinoline (9.6 g, 19 mmol), 4,4,5,5-tetramethyl-2-(4,4,1,5,5-tetramethyl-1,3,2-dioxoboran-2-yl)-1,2,2-dioxyborane (10 g, 20 mmol), AcOK (5.78 g, 60 mmol) in toluene (110 mL) was added Pd(dppf)Cl ₂ (1.44 g, 2 mmol). The mixture was degassed and stirred at 130 °C for 3 hours. The reaction mixture was filtered and concentrated to give a residue. To the residue was added EtOAc (100 mL) and water (80 mL). The organic phase was washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give a residue. The residue was purified by column chromatography (SiO ₂ ) and eluted with 30-40% ethyl acetate in petroleum ether to give (2-fluoro-6-methoxy-8-(4,4,5,5-tetramethyl-1,3,2-dioxoboran-2-yl)isoquinolin-1-yl)ethynyl)triisopropylsilane (4.4 g, 48% yield). LC-MS (ESI): m/z 484 [M+H] ⁺ .

8- Fluoro -3- methoxyisoquinoline -1- pinacol borate

Step A: To a solution of methyl 2-fluoro-6-methylbenzoate (16.8 g, 100 mmol) in CCl ₄ (130 mL) was added NBS (26.7 g, 150 mmol) and benzoyl peroxide (1.46 g, 7.8 mmol). The reaction mixture was stirred at 85°C under nitrogen atmosphere for 3 hours. The crude product was filtered and washed with toluene. The volatiles were evaporated, the residue was dissolved in DMF (150 mL), and sodium cyanide (9.80 g) was added. The resulting mixture was heated at 60°C for 5 hours. The reaction mixture was cooled to room temperature, treated with water (50 mL), and extracted with ethyl acetate (3 x 50 mL). The combined organics were washed with aqueous saline solution (50 mL) and dried over anhydrous sodium sulfate. The solution was filtered and concentrated under reduced pressure to give the crude product. Elution with 30-40% ethyl acetate in petroleum ether gave methyl 2-fluoro-6-(cyanomethyl)benzoate (13.1 g) [M+H] ⁺ . LC-MS (ESI): m/z 194.

Step B: To a solution of methyl 2-fluoro-6-(cyanomethyl)benzoate (9.7 g, 50 mmol) in methanol (140 mL) was added sodium methoxide (25% in methanol, 50 mL). The resulting mixture was heated at 70 °C for 4 h. The reaction mixture was cooled to room temperature and acidified using 1N HCl. The precipitated solid was filtered, washed with water and dried under vacuum to give 8-fluoro-3-methoxy-2H-isoquinolin-l-one (9.74 g, 101% yield). LC-MS (ESI): m/z 194 [M+H] ⁺ .

Step C: To a solution of 8-fluoro-3-methoxy-2H-isoquinolin-l-one (3.86 g, 20 mmol), DIEA (7.7 g, 60 mmol) in DCM (120 mL) was added Tf ₂ O (8.41 g, 30 mmol), and the mixture was stirred at -40 °C for 0.5 h. The reaction mixture was diluted with ice water (50 mL), and then extracted with DCM (30 mL). The combined organic phases were dried over Na ₂ SO ₄ and concentrated to dryness. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate = 1/0 to 60/1) to give 1-trifluoromethanesulfonyloxy-8-fluoro-3-methoxyisoquinoline (6.24 g, 96% yield). LC-MS(ESI): m/z 326[M+H] ⁺ .

Step D: To a mixture of 1-trifluoromethanesulfonyloxy-8-fluoro-3-methoxyisoquinoline (6.19 g, 19 mmol), 4,4,5,5-tetramethyl-2-(4,4,1,5,5-tetramethyl-1,3,2-dioxoboran-2-yl)-1,2,2-dioxyborane (10 g, 20 mmol), AcOK (5.78 g, 60 mmol) in toluene (110 mL) was added Pd(dppf)Cl ₂ (1.44 g, 2 mmol). The mixture was degassed and stirred at 130 °C for 3 hours. The reaction mixture was filtered and concentrated to obtain a residue. To the residue was added EtOAc (100 mL) and water (80 mL). The organic phase was washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give a residue. The residue was purified by column chromatography (SiO ₂ ) eluting with 30-40% ethyl acetate in petroleum ether to give 8-fluoro-3-methoxyisoquinoline-1-pinacol borate (3.57 g, 62% yield) [M+H] ⁺ . LC-MS (ESI): m/z 304.

8- Chloro -3- methoxyisoquinoline -1- pinacol borate

Step A: To a solution of methyl 2-chloro-6-methylbenzoate (18.4 g, 100 mmol) in CCl ₄ (130 mL) was added NBS (26.7 g, 150 mmol) and benzoyl peroxide (1.46 g, 7.8 mmol). The reaction mixture was stirred at 85°C under nitrogen atmosphere for 3 hours. The crude product was filtered and washed with toluene. The volatiles were evaporated, the residue was dissolved in DMF (150 mL), and sodium cyanide (9.80 g) was added. The resulting mixture was heated at 60°C for 5 hours. The reaction mixture was cooled to room temperature, treated with water (50 mL), and extracted with ethyl acetate (3 x 50 mL). The combined organics were washed with saline solution (50 mL) and dried over anhydrous sodium sulfate. The solution was filtered and concentrated under reduced pressure to give the crude product. Elution with 30-40% ethyl acetate in petroleum ether gave methyl 2-chloro-6-(cyanomethyl)benzoate (14.0 g). LC-MS (ESI): m/z 210 [M+H] ⁺ .

Step B: To a solution of methyl 2-chloro-6-(cyanomethyl)benzoate (9.7 g, 50 mmol) in methanol (140 mL) was added sodium methoxide (25% in methanol, 50 mL). The resulting mixture was heated at 70 °C for 4 h. The reaction mixture was cooled to room temperature and acidified using 1N HCl. The precipitated solid was filtered, washed with water and dried under vacuum to give 8-chloro-3-methoxy-2H-isoquinolin-l-one (10.76 g, 103% yield). LC-MS (ESI): m/z 210 [M+H] ⁺ .

Step C: To a solution of 8-chloro-3-methoxy-2H-isoquinolin-l-one (4.18 g, 20 mmol), DIEA (7.7 g, 60 mmol) in DCM (120 mL) was added Tf ₂ O (8.41 g, 30 mmol), and the mixture was stirred at -40 °C for 0.5 h. The reaction mixture was diluted with ice water (50 mL), and then extracted with DCM (30 mL). The combined organic phases were dried over Na ₂ SO ₄ and concentrated to dryness. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate = 1/0 to 60/1) to give 1-trifluoromethanesulfonyloxy-8-chloro-3-methoxyisoquinoline (6.61 g, 97% yield). LC-MS(ESI): m/z 342[M+H] ⁺ .

Step D: To a mixture of 1-trifluoromethanesulfonyloxy-8-chloro-3-methoxyisoquinoline (6.48 g, 19 mmol), 4,4,5,5-tetramethyl-2-(4,4,1,5,5-tetramethyl-1,3,2-dioxoboran-2-yl)-1,2,2-dioxyborane (10 g, 20 mmol), AcOK (5.78 g, 60 mmol) in toluene (110 mL) was added Pd(dppf)Cl ₂ (1.44 g, 2 mmol). The mixture was degassed and stirred at 130 °C for 3 hours. The reaction mixture was filtered and concentrated to obtain a residue. To the residue was added EtOAc (100 mL) and water (80 mL). The organic phase was washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give a residue. The residue was purified by column chromatography (SiO ₂ ) and eluted with 30-40% ethyl acetate in petroleum ether to give 8-chloro-3-methoxyisoquinoline-1-pinacol borate (4.6 g, 76% yield). LC-MS (ESI): m/z 320 [M+H] ⁺ .

7- Fluoro -8- chloro -3- methoxyisoquinoline -1- pinacol borate

Step A: To a solution of methyl 2-chloro-6-methylbenzoate (20.2 g, 100 mmol) in CCl ₄ (130 mL) was added NBS (26.7 g, 150 mmol) and benzoyl peroxide (1.46 g, 7.8 mmol). The reaction mixture was stirred at 85°C under nitrogen atmosphere for 3 hours. The crude product was filtered and washed with toluene. The volatiles were evaporated, the residue was dissolved in DMF (150 mL), and sodium cyanide (9.80 g) was added. The resulting mixture was heated at 60°C for 5 hours. The reaction mixture was cooled to room temperature, treated with water (50 mL), and extracted with ethyl acetate (3 x 50 mL). The combined organics were washed with saline solution (50 mL) and dried over anhydrous sodium sulfate. The solution was filtered and concentrated under reduced pressure to give the crude product. Elution with 30-40% ethyl acetate in petroleum ether gave methyl 2-chloro-6-(cyanomethyl)benzoate (14.75 g). LC-MS (ESI): m/z 228 [M+H] ⁺ .

Step B: To a solution of methyl 2-chloro-6-(cyanomethyl)benzoate (11.35 g, 50 mmol) in methanol (140 mL) was added sodium methoxide (25% in methanol, 50 mL). The resulting mixture was heated at 70 °C for 4 h. The reaction mixture was cooled to room temperature and acidified using 1N HCl. The precipitated solid was filtered, washed with water and dried under vacuum to give 8-chloro-3-methoxy-2H-isoquinolin-l-one (11.58 g, 102% yield). LC-MS (ESI): m/z 228 [M+H] ⁺ .

Step C: To a solution of 8-chloro-7-fluoro-3-methoxy-2H-isoquinolin-l-one (4.54 g, 20 mmol), DIEA (7.7 g, 60 mmol) in DCM (120 mL) was added Tf ₂ O (8.41 g, 30 mmol), and the mixture was stirred at -40 °C for 0.5 h. The reaction mixture was diluted with ice water (50 mL), and then extracted with DCM (30 mL). The combined organic phases were dried over Na ₂ SO ₄ and concentrated to dryness. The residue was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate = 1/0 to 60/1) to give 1-trifluoromethanesulfonyloxy-8-chloro-7-fluoro-3-methoxyisoquinoline (6.82 g, 95% yield). LC-MS(ESI): m/z 360[M+H] ⁺ .

Step D: To a mixture of 1-trifluoromethanesulfonyloxy-8-chloro-7-fluoro-3-methoxyisoquinoline (6.82 g, 19 mmol), 4,4,5,5-tetramethyl-2-(4,4,1,5,5-tetramethyl-1,3,2-dioxoborolan-2-yl)-1,2,2-dioxyborane (10 g, 20 mmol), AcOK (5.78 g, 60 mmol) in toluene (110 mL) was added Pd(dppf)Cl ₂ (1.44 g, 2 mmol). The mixture was degassed and stirred at 130 °C for 3 hours. The reaction mixture was filtered and concentrated to obtain a residue. To the residue was added EtOAc (100 mL) and water (80 mL). The organic phase was washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give a residue. The residue was purified by column chromatography (SiO ₂ ) and eluted with 30-40% ethyl acetate in petroleum ether to give 8-chloro-7-fluoro-3-methoxyisoquinoline-1-pinacol borate (4.74 g, 74% yield). LC-MS (ESI): m/z 338 [M+H] ⁺ .

8- Methyl -3- methoxyisoquinoline -1- pinacol borate

Step A: To a solution of methyl 2,6-methylbenzoate (16.4 g, 100 mmol) in CCl ₄ (130 mL) was added NBS (26.7 g, 150 mmol) and benzoyl peroxide (1.46 g, 7.8 mmol). The reaction mixture was stirred at 85°C under nitrogen atmosphere for 3 hours. The crude product was filtered and washed with toluene. The volatiles were evaporated, the residue was dissolved in DMF (150 mL), and sodium cyanide (9.80 g) was added. The resulting mixture was heated at 60°C for 5 hours. The reaction mixture was cooled to room temperature, treated with water (50 mL), and extracted with ethyl acetate (3 x 50 mL). The combined organics were washed with brine solution (50 mL) and dried over anhydrous sodium sulfate. The solution was filtered and concentrated under reduced pressure to give the crude product. Elution with 30-40% ethyl acetate in petroleum ether gave methyl 2-methyl-6-(cyanomethyl)benzoate (10.96 g). LC-MS (ESI): m/z 190 [M+H] ⁺ .

Step B: To a solution of methyl 2-methyl-6-(cyanomethyl)benzoate (9.45 g, 50 mmol) in methanol (140 mL) was added sodium methoxide (25% in methanol, 50 mL). The resulting mixture was heated at 70 °C for 4 h. The reaction mixture was cooled to room temperature and acidified using 1N HCl. The precipitated solid was filtered, washed with water and dried under vacuum to give 8-chloro-3-methoxy-2H-isoquinolin-l-one (9.83 g, 104% yield). LC-MS (ESI): m/z 190 [M+H] ⁺ .

Step C: To a solution of 8-methyl-3-methoxy-2H-isoquinolin-l-one (3.78 g, 20 mmol), DIEA (7.7 g, 60 mmol) in DCM (120 mL) was added Tf ₂ O (8.41 g, 30 mmol), and the mixture was stirred at -40 °C for 0.5 h. The reaction mixture was diluted with ice water (50 mL), and then extracted with DCM (30 mL). The combined organic phases were dried over Na ₂ SO ₄ and concentrated to dryness to give 1-trifluoromethanesulfonyloxy-8-fluoro-3-methoxyisoquinoline (6.16 g, 96% yield). LC-MS (ESI): m/z 322 [M+H] ⁺ .

Step D: To a mixture of 1-trifluoromethanesulfonyloxy-8-methyl-3-methoxyisoquinoline (6.06 g, 19 mmol), 4,4,5,5-tetramethyl-2-(4,4,1,5,5-tetramethyl-1,3,2-dioxoborolan-2-yl)-1,2,2-dioxyborane (10 g, 20 mmol), AcOK (5.78 g, 60 mmol) in toluene (110 mL) was added Pd(dppf)Cl ₂ (1.44 g, 2 mmol). The mixture was degassed and stirred at 130 °C for 3 hours. The reaction mixture was filtered and concentrated to obtain a residue. To the residue was added EtOAc (100 mL) and water (80 mL). The organic phase was washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give a residue. The residue was purified by column chromatography (SiO ₂ ) and eluted with 30-40% ethyl acetate in petroleum ether to give 8-methyl-3-methoxyisoquinoline-1-pinacol borate (4.2 g, 74% yield). LC-MS (ESI): m/z 300 [M+H] ⁺ .

7- Fluoro -8 - ethyl -3- methoxyisoquinoline -1- pinacol borate

Step A: To a stirred solution of methyl 2-iodo-3-fluoro-6-(cyanomethyl)benzoate (44.5 g, 139.5 mmol) in toluene (240 mL) and water (60 mL) was added ethylboric acid (15.4 g, 209 mmol), potassium phosphate (59.2 g, 279 mmol) and tricyclohexylphosphine (3.9 g, 13.9 mmol) and the reaction mixture was stirred at 100 °C for 3 hours. After the reaction was completed, the reaction was cooled to room temperature, quenched with water and extracted with ethyl acetate (2*500 mL). The organics were combined and washed with brine solution (300 mL), dried over Na ₂ SO ₄ , filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate = 1/0 to 60/1) to give methyl 2-ethyl-3-fluoro-6-(cyanomethyl)benzoate (20.79 g, 65%). LC-MS (ESI): m/z 221 [M+H] ⁺ .

Step B: To a solution of methyl 2-ethyl-3-fluoro-6-(cyanomethyl)benzoate (11.05 g, 50 mmol) in methanol (140 mL) was added sodium methoxide (25% in methanol, 50 mL). The resulting mixture was heated at 70 °C for 4 h. The reaction mixture was cooled to room temperature and acidified using 1N HCl. The precipitated solid was filtered, washed with water and dried under vacuum to give 8-ethyl-7-fluoro-3-methoxy-2H-isoquinolin-l-one (11.27 g, 102% yield). LC-MS (ESI): m/z 221 [M+H] ⁺ .

Step C: To a solution of 8-ethyl-7-fluoro-3-methoxy-2H-isoquinolin-l-one (4.4 g, 20 mmol), DIEA (7.7 g, 60 mmol) in DCM (120 mL) was added Tf ₂ O (8.41 g, 30 mmol), and the mixture was stirred at -40 °C for 0.5 h. The reaction mixture was diluted with ice water (50 mL), and then extracted with DCM (30 mL). The combined organic phases were dried over Na ₂ SO ₄ and concentrated to dryness to give 1-trifluoromethanesulfonyloxy-8-ethyl-7-fluoro-3-methoxyisoquinoline (6.84 g, 97%). LC-MS (ESI): m/z 354 [M+H] ⁺ .

Step D: To a mixture of 1-trifluoromethanesulfonyloxy-8-ethyl-7-fluoro-3-methoxyisoquinoline (6.7 g, 19 mmol), 4,4,5,5-tetramethyl-2-(4,4,1,5,5-tetramethyl-1,3,2-dioxoborolan-2-yl)-1,2,2-dioxyborane (10 g, 20 mmol), AcOK (5.78 g, 60 mmol) in toluene (110 mL) was added Pd(dppf)Cl ₂ (1.44 g, 2 mmol). The mixture was degassed and stirred at 130 °C for 3 hours. The reaction mixture was filtered and concentrated to obtain a residue. To the residue was added EtOAc (100 mL) and water (80 mL). The organic phase was washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give a residue. The residue was purified by column chromatography (SiO ₂ ) and eluted with 30-40% ethyl acetate in petroleum ether to give 7-fluoro-8-ethyl-3-methoxyisoquinoline-1-pinacol borate (3.52 g, 56% yield). LC-MS (ESI): m/z 332 [M+H] ⁺ .

7- Fluoro -8- isopropyl -3- methoxyisoquinoline -1- pinacol borate

Step A: To a stirred solution of methyl 2-iodo-3-fluoro-6-(cyanomethyl)benzoate (44.5 g, 139.5 mmol) in toluene (240 mL) and water (60 mL) was added isopropylboric acid (18.3 g, 209 mmol), potassium phosphate (59.2 g, 279 mmol) and tricyclohexylphosphine (3.9 g, 13.9 mmol) and the reaction mixture was stirred at 100 °C for 3 hours. After the reaction was completed, the reaction was cooled to room temperature, quenched with water and extracted with ethyl acetate (2*500 mL). The organics were combined and washed with brine solution (300 mL), dried over Na ₂ SO ₄ , filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate = 1/0 to 60/1) to give methyl 2-isopropyl-3-fluoro-6-(cyanomethyl)benzoate (24.01 g, 52%). LC-MS (ESI): m/z 236 [M+H] ⁺ .

Step B: To a solution of methyl 2-isopropyl-3-fluoro-6-(cyanomethyl)benzoate (11.75 g, 50 mmol) in methanol (140 mL) was added sodium methoxide (25% in methanol, 50 mL). The resulting mixture was heated at 70 °C for 4 h. The reaction mixture was cooled to room temperature and acidified using 1N HCl. The precipitated solid was filtered, washed with water and dried under vacuum to give 8-isopropyl-7-fluoro-3-methoxy-2H-isoquinolin-l-one (11.98 g, 102% yield). LC-MS (ESI): m/z 320 [M+H] ⁺ .

Step C: To a solution of 8-isopropyl-7-fluoro-3-methoxy-2H-isoquinolin-l-one (7.4 g, 20 mmol), DIEA (7.7 g, 60 mmol) in DCM (120 mL) was added Tf ₂ O (8.41 g, 30 mmol), and the mixture was stirred at -40 °C for 0.5 h. The reaction mixture was diluted with ice water (50 mL), and then extracted with DCM (30 mL). The combined organic phases were dried over Na ₂ SO ₄ and concentrated to dryness to give 1-trifluoromethanesulfonyloxy-8-isopropyl-7-fluoro-3-methoxyisoquinoline (6.32 g, 99% yield). LC-MS (ESI): m/z 320 [M+H] ⁺ .

Step D: To a mixture of 1-trifluoromethanesulfonyloxy-8-isopropyl-7-fluoro-3-methoxyisoquinoline (6.06 g, 19 mmol), 4,4,5,5-tetramethyl-2-(4,4,1,5,5-tetramethyl-1,3,2-dioxoborolan-2-yl)-1,2,2-dioxyborane (10 g, 20 mmol), AcOK (5.78 g, 60 mmol) in toluene (110 mL) was added Pd(dppf)Cl ₂ (1.44 g, 2 mmol). The mixture was degassed and stirred at 130 °C for 3 hours. The reaction mixture was filtered and concentrated to obtain a residue. To the residue was added EtOAc (100 mL) and water (80 mL). The organic phase was washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give a residue. The residue was purified by column chromatography (SiO ₂ ) and eluted with 30-40% ethyl acetate in petroleum ether to give 7-fluoro-8-isopropyl-3-methoxyisoquinoline-1-pinacol borate (3.14 g, 48% yield). LC-MS (ESI): m/z 346 [M+H] ⁺ .

7- Fluoro -8- cyclopropyl -3- methoxyisoquinoline -1- pinacol borate

Step A: To a stirred solution of methyl 2-iodo-3-fluoro-6-(cyanomethyl)benzoate (44.5 g, 139.5 mmol) in toluene (240 mL) and water (60 mL) was added isopropylboric acid (18.3 g, 209 mmol), potassium phosphate (59.2 g, 279 mmol) and tricyclohexylphosphine (3.9 g, 13.9 mmol) and the reaction mixture was stirred at 100 °C for 3 hours. After the reaction was completed, the reaction was cooled to room temperature, quenched with water and extracted with ethyl acetate (2*500 mL). The organics were combined and washed with brine solution (300 mL), dried over Na ₂ SO ₄ , filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (SiO ₂ , petroleum ether/ethyl acetate = 1/0 to 60/1) to give methyl 2-cyclopropyl-3-fluoro-6-(cyanomethyl)benzoate (14.95 g, 46%). LC-MS (ESI): m/z 234 [M+H] ⁺ .

Step B: To a solution of methyl 2-cyclopropyl-3-fluoro-6-(cyanomethyl)benzoate (11.65 g, 50 mmol) in methanol (140 mL) was added sodium methoxide (25% in methanol, 50 mL). The resulting mixture was heated at 70 °C for 4 h. The reaction mixture was cooled to room temperature and acidified using 1N HCl. The precipitated solid was filtered, washed with water and dried under vacuum to give 8-cyclopropyl-7-fluoro-3-methoxy-2H-isoquinolin-l-one ((11.76 g, 101% yield). LC-MS (ESI): m/z 234 [M+H] ⁺ .

Step C: To a solution of 8-cyclopropyl-7-fluoro-3-methoxy-2H-isoquinolin-l-one (4.66 g, 20 mmol), DIEA (7.7 g, 60 mmol) in DCM (120 mL) was added Tf ₂ O (8.41 g, 30 mmol), and the mixture was stirred at -40 °C for 0.5 h. The reaction mixture was diluted with ice water (50 mL), and then extracted with DCM (30 mL). The combined organic phases were dried over Na ₂ SO ₄ and concentrated to dryness to give 1-trifluoromethanesulfonyloxy-8-cyclopropyl-7-fluoro-3-methoxyisoquinoline (7.08 g, 97% yield). LC-MS (ESI): m/z 366[M+H] ⁺ .

Step D: To a mixture of 1-trifluoromethanesulfonyloxy-8-cyclopropyl-7-fluoro-3-methoxyisoquinoline (6.93 g, 19 mmol), 4,4,5,5-tetramethyl-2-(4,4,1,5,5-tetramethyl-1,3,2-dioxoboran-2-yl)-1,2,2-dioxoborane (10 g, 20 mmol), AcOK (5.78 g, 60 mmol) in toluene (110 mL) was added Pd(dppf)Cl ₂ (1.44 g, 2 mmol). The mixture was degassed and stirred at 130 °C for 3 hours. The reaction mixture was filtered and concentrated to obtain a residue. To the residue was added EtOAc (100 mL) and water (80 mL). The organic phase was washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated to give a residue. The residue was purified by column chromatography (SiO2) and eluted with 30-40% ethyl acetate in petroleum ether to give 7-fluoro-8-cyclopropyl-3-methoxyisoquinoline-1-pinacol borate (2.87 g, 44% yield). LC-MS (ESI): m/z 344 [M+H] ⁺ .

1,3,7- Triazaspiro [4.5] decan -2- one

Step A: To a solution of 7-Cbz-2,4-dioxo-1,3,7-triazaspiro[4.5]decane (2.00 g 6.6 mmol) in THF (40 mL) was added BH ₃ .THF (1 M, 39.6 mL) at 0°C and the reaction was stirred at 80°C for 2 hours. MeOH (40 mL) was then slowly added at 0°C and the mixture was stirred at 20°C for 0.5 hours and then concentrated. The residue was purified by reverse phase flash chromatography to give 7-Cbz-2-oxo-1,3,7-triazaspiro[4.5]decane (1.57 g, 80%) as a yellow solid. LC-MS(ESI): m/z 290[M+H] ⁺ .

Step B: A mixture of 7-Cbz-2-oxo-1,3,7-triazaspiro[4.5]decane (1.63 g, 5 mmol) and Pd/C (330 mg, 10% purity) in MeOH (100 mL) was added and stirred at 25°C under H ₂ (15 psi) for 1 hour. The reaction was then filtered and concentrated under reduced pressure and purified by column chromatography (SiO ₂ ) to give 2-oxo-1,3,7-triazaspiro[4.5]decane (0.728 g, 94%). LC-MS (ESI): m/z 156[M+H] ⁺ .

2- Thia -1,3,7- triazaspiro [4.5] decan -2,2- one

Step A: A solution of sulfonamide (4.2 g, 44 mmol) in pyridine (100 mL) was stirred at 120° C. for 10 min, then N-Cbz-3-amino-3-(aminomethyl)piperidine (5 g, 22 mmol) was added to the solution, and the mixture was stirred at 120° C. for 16 h. The mixture was concentrated in vacuo, and then poured into 200 mL of H ₂ O and extracted with EA (100 mL×3). The combined organic layers were washed with water (80 mL×2) and brine (80 mL x 2), dried over Na ₂ SO ₄ and concentrated in vacuo. Purification by column chromatography (SiO ₂ ) gave yellow solid 7-Cbz-2-thia-1,3,7-triazaspiro[4.5]decane-2,2-one (3.21 g, 45%). LC-MS (ESI): m/z 326[M+H] ⁺ .

Step B: A mixture of 7-Cbz-2-thia-1,3,7-triazaspiro[4.5]decane-2,2-one (1.63 g, 5 mmol) and Pd/C (330 mg, 10% purity) in MeOH (100 mL) was added and stirred at 25°C under H ₂ (15 psi) for 1 hour. The reaction was then filtered and concentrated under reduced pressure and purified by column chromatography (SiO ₂ ) to give 2-thia-1,3,7-triazaspiro[4.5]decane-2,2-one (0.908 g, 95%). LC-MS (ESI): m/z 192[M+H] ⁺ .

(R)-2- Thia -1,3,7- triazaspiro [4.5] decan -2,2- one

Step A: A solution of sulfonamide (4.2 g, 44 mmol) in pyridine (100 mL) was stirred at 120° C. for 10 min, then (3R)-N-Cbz-3-amino-3-(aminomethyl)piperidine (5 g, 22 mmol) was added to the solution, and the mixture was stirred at 120° C. for 16 h. The mixture was concentrated in vacuo, and then poured into 200 mL of H ₂ O and extracted with EA (100 mL×3). The combined organic layers were washed with water (80 mL×2) and brine (80 mL x 2), dried over Na ₂ SO ₄ and concentrated in vacuo. Purification by column chromatography (SiO ₂ ) gave a yellow solid (R)-7-Cbz-2-thia-1,3,7-triazaspiro[4.5]decane-2,2-one (3.0 g, 42%). LC-MS (ESI): m/z 326[M+H] ⁺ .

Step B: Add a mixture of (R)-7-Cbz-2-thia-1,3,7-triazaspiro[4.5]decane-2,2-one (1.63 g, 5 mmol) in MeOH (100 mL) and Pd/C (330 mg, 10% purity) and stir at 25°C under H ₂ (15 psi) for 1 hour. The reaction was then filtered and concentrated under reduced pressure and purified by column chromatography (SiO ₂ ) to give (R)-2-thia-1,3,7-triazaspiro[4.5]decane-2,2-one (0.888 g, 93%). LC-MS (ESI): m/z 192[M+H] ⁺ .

(5R)-2- Methyl -1,3,7- triazaspiro -2- phosphaspiro [4.5] decan -2- one

Step A: A solution of methylphosphonodiamine (4.14 g, 44 mmol) in bromobenzene (100 mL) was stirred at 120° C. for 10 min, then (3R)-N-Cbz-3-amino-3-(aminomethyl)piperidine (5 g, 22 mmol) was added to the solution, and the mixture was stirred at 120° C. for 16 h. The mixture was concentrated in vacuo, and then the mixture was poured into 200 mL of H ₂ O and extracted with EA (100 mL×3). The combined organic layers were washed with water (80 mL×2) and brine (80 mL x 2), dried over Na ₂ SO ₄ and concentrated in vacuo, and purified by column chromatography (SiO ₂ ) to give a yellow solid 1-Cbz-(5R)-2-methyl-1,3,7-triazaspiro-2-phosphaspiro[4.5]decan-2-one (2.7 g, 38%). LC-MS (ESI): m/z 324[M+H] ⁺ .

Step B: Add 1-Cbz-(5R)-2-methyl-1,3,7-triazaspiro-2-phosphaspiro[4.5]decan2-one (1.63 g, 5 mmol) in MeOH (100 mL) and a mixture of Pd/C (330 mg, 10% purity) and stir at 25°C under H ₂ (15 psi) for 1 hour. The reaction was then filtered and concentrated under reduced pressure and purified by column chromatography (SiO ₂ ) to give (5R)-2-methyl-1,3,7-triazaspiro-2-phosphaspiro[4.5]decan2-one (0.917 g, 96%). LC-MS (ESI): m/z 192[M+H] ⁺ .

N,N-Dimethyl-5,6,7,8-tetrahydro-4H-pyrazolo[1,5-a][1,4]diazepine-2-carboxamide To a solution of 5-(tert-butyloxycarbonyl)-5,6,7,8-tetrahydro-4H-pyrazolo[1,5-a][1,4]diazepine-2-carboxylic acid (10 g) and dimethylamine (35.5 mL) in DCM (100 mL) were added HATU (20.3 g) and N,N-diisopropylethylamine (13.8 g, 18.6 mL). The mixture was stirred at 20 °C for 1 hour. The mixture was concentrated under vacuum. The residue was recrystallized from ethanol to give N-Boc-N,N-dimethyl-5,6,7,8-tetrahydro-4H-pyrazolo[1,5-a][1,4]diazepine-2-carboxamide (9.17 g). LC-MS (ESI): m/z 309 [M+H] ⁺ .

A mixture of N-Boc-N,N-dimethyl-5,6,7,8-tetrahydro-4H-pyrazolo[1,5-a][1,4]diazepine-2-carboxamide (9 g) and HCl/dioxane (95 mL) in MeCN (95 mL) was stirred at 0°C for 1 hour. The mixture was concentrated under vacuum. The pH of the residue was adjusted to 8 with saturated sodium bicarbonate (4 mL). Filtration gave N,N-dimethyl-5,6,7,8-tetrahydro-4H-pyrazolo[1,5-a][1,4]diazepine-2-carboxamide (5.13 g). LC-MS (ESI): m/z 209 [M+H] ⁺ .

Embodiment 1

Step A: Mix 2,4,7-trichloropyrido[2,3-d]pyrimidine (2.52 g, 10 mmol), 2-oxo-1,3,7-triazaspiro[4.5]decane (1.70 g, 11 mmol), potassium carbonate (2.07 g, 15 mmol), catalytic amount of potassium iodide and DMF (60 mL), heat to 120°C, and stir for 4 hours. Cool to room temperature, evaporate under reduced pressure to obtain yellow solid 2,7-dichloro-4-(2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)pyrido[2,3-d]pyrimidine (5.50 g, 78%), LC/MS(ESI): m/z =354[M+H] ⁺ .

Step B: Mix 2,7-dichloro-4-(2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)pyrido[2,3-d]pyrimidine (353 mg, 1 mmol), 1,2,3,5,6,7-hexahydropyrrolizin-7-yl]methanol (155 mg, 1.1 mmol), potassium carbonate (0.12 mg, 1.4 mmol), a catalytic amount of potassium iodide and DMF (8 mL), heat to 120°C, and stir for 4 hours. The mixture was cooled to room temperature and evaporated under reduced pressure to give yellow solid 7-chloro-2-((tetrahydro-1H-pyrinyl-7a(5H)-yl)methoxy)-4-(2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)pyridin[2,3-d]pyrimidine (0.375 g, 82%). LC/MS(ESI): m/z =458[M+H] ⁺ .

Step C: 7-chloro-2((tetrahydro-1H-pyrinylcyclo-7a(5H)-yl)methoxy)-4-(2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)pyridin[2,3-d]pyrimidine (92 mg, 0.2 mmol), 3-methoxy-8-ethyl-7-fluoroisoquinolinylboronic acid pinacol ester (66.4 mg, 0.2 mmol), [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (16 mg, 0.022 mmol), K ₃ PO ₄ , THF (5 mL) and water (1 mL) were mixed, then heated to 60° C. under reflux and stirred for 16 hours. The reaction was cooled to room temperature and stirred overnight to give a light yellow precipitate. The reaction mixture was diluted with water (2 mL) and the solid was collected by filtration. The crude product was column chromatographed to give 7-(3-methoxy-8-ethyl-7-fluoroisoquinolin-1-yl)-4-(2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)-2-((2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[2,3-d]pyrimidine (72 mg, 58% yield). LC/MS(ESI): m/z =627.3[M+H] ⁺ .

Step D: 7-(3-methoxy-8-ethyl-7-fluoroisoquinolin-1-yl)-4-(2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)-2-((tetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[2,3-d]pyrimidine (63 mg, 0.1 mmol) was dissolved in DCM (2 mL) at -78°C, BBr ₃ ((0.4 mL, 1M in DCM , 0.4 mmol) was added, and then the mixture was heated to room temperature and stirred for 5 h, then cooled to -78°C, adjusted to Ph = 7 with 1N NaOH, extracted with EA, and then concentrated by filtration under reduced pressure and purified by preparative HPLC to give compound 1 (32 mg, yield 56%). LC/MS(ESI): m/z =613.3[M+H] ⁺ .

Embodiment 2

Step A: Mix 2,7-dichloro-4-(2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)pyridin[2,3-d]pyrimidine (250 mg, 1 mmol), N-methyl-L-prolinol (127 mg, 1.1 mmol), potassium carbonate (0.12 mg, 1.4 mmol), catalytic amount of potassium iodide and DMF (8 mL), heat to 120°C, and stir for 4 hours. Cool to room temperature, and evaporate the yellow solid 7-chloro-2(N-methylpyrrolidin-2-methoxy)-4-(2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)pyridin[2,3-d]pyrimidine (367 mg) under reduced pressure. LC/MS(ESI): m/z =433.2[M+H] ⁺ .

Step B: 7-chloro-2(N-methylpyrrolidin-2-methoxy)-4-(2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)pyridin[2,3-d]pyrimidine (87 mg, 0.2 mmol), 3-methoxy-8-ethyl-7-fluoroisoquinolinylboronic acid pinacol ester (66.4 mg, 0.2 mmol), [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (16 mg, 0.022 mmol), K ₃ PO ₄ , THF (5 mL) and water (1 mL) were mixed, then heated to 60° C. under reflux and stirred for 16 hours. The reaction was cooled to room temperature and stirred overnight to give a light yellow precipitate. The reaction mixture was diluted with water (2 mL) and the solid was collected by filtration. The crude product was purified by column chromatography to give 7-(3-methoxy-8-ethyl-7-fluoroisoquinolin-1-yl)-4-(2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)-2-((N-methylpyrrolidin-2-methoxy)pyridin[2,3-d]pyrimidine (89 mg). LC/MS(ESI): m/z =600.3[M+H] ⁺ .

Step C: 7-(3-methoxy-8-ethyl-7-fluoroisoquinolin-1-yl)-4-(2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)-2-((N-methylpyrrolidin-2-methoxy)pyridin[2,3-d]pyrimidine (60 mg, 0.1 mmol) was dissolved in DCM (2 mL) at -78°C, BBr ₃ ((0.4 mL, 1M in DCM, 0.4 mmol) was added, and then the mixture was heated to room temperature and stirred for 5 h, then cooled to -78°C, adjusted to Ph = 7 with 1N NaOH, extracted with EA, and then filtered under reduced pressure and concentrated and purified by preparative HPLC to give compound 2 (39 mg, yield 67%). LC/MS(ESI): m/z =587.3[M+H] ⁺ .

Embodiment 3

Step A: 2,7-dichloro-4-(2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)pyridinium[2,3-d]pyrimidine (250 mg, 1 mmol), 1-(((dimethylamino)cyclopropyl)methanol (148 mg, 1.1 mmol), potassium carbonate (0.12 mg, 1.4 mmol), catalytic amount of potassium iodide and DMF (8 mL) were mixed, heated to 120°C, and stirred for 4 hours. The mixture was cooled to room temperature and evaporated under reduced pressure to obtain 7-chloro-2(1-(((dimethylamino)cyclopropyl)methoxy)-4-(2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)pyridinium[2,3-d]pyrimidine (337 mg) as a yellow solid. LC/MS(ESI): m/z =433.2[M+H] ⁺ .

Step B: 7-chloro-2-(1-((dimethylamino)cyclopropyl)methoxy)-4-(2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)pyridin[2,3-d]pyrimidine (87 mg, 0.2 mmol), 3-methoxy-8-ethyl-7-fluoroisoquinolinylboronic acid pinacol ester (66.4 mg, 0.2 mmol), [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (16 mg, 0.022 mmol), K ₃ PO ₄ , THF (5 mL) and water (1 mL) were mixed, then heated to 60° C. under reflux and stirred for 16 hours. The reaction was cooled to room temperature and stirred overnight to give a light yellow precipitate. The reaction mixture was diluted with water (2 mL) and the solid was collected by filtration. The crude product was chromatographed on a methanol column to give 7-(3-methoxy-8-ethyl-7-fluoroisoquinolin-1-yl)-4-(2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)-2-(1-((dimethylamino)cyclopropyl)methoxy)pyridin[2,3-d]pyrimidine (69 mg, yield 58%). LC/MS(ESI): m/z =600.3[M+H] ⁺ .

Step C: 7-(3-methoxy-8-ethyl-7-fluoroisoquinolin-1-yl)-4-(2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)-2-(1-((dimethylamino)cyclopropyl)methoxy)pyridin[2,3-d]pyrimidine (60 mg, 0.1 mmol) was dissolved in DCM (2 mL) at -78°C, BBr ₃ ((0.4 mL, 1M in DCM, 0.4 mmol) was added, and then the mixture was heated to room temperature and stirred for 5 h, then cooled to -78°C, adjusted to Ph = 7 with 1N NaOH, extracted with EA, and then concentrated by filtration under reduced pressure and purified by preparative HPLC to give compound 3 (34 mg, yield 58%). LC/MS(ESI): m/z =587.2[M+H] ⁺ .

Embodiment 4

Step A: Mix 2,7-dichloro-4-(2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)pyridin[2,3-d]pyrimidine (250 mg, 1 mmol), 1-(pyrrol-1-yl)cyclopropyl)methanol (155 mg, 1.1 mmol), potassium carbonate (0.12 mg, 1.4 mmol), catalytic amount of potassium iodide and DMF (8 mL), heat to 120°C, and stir for 4 hours. Cool to room temperature, evaporate under reduced pressure to obtain yellow solid 7-chloro-2(1-(pyrrol-1-yl)cyclopropyl)methoxy)-4-(2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)pyridin[2,3-d]pyrimidine (357 mg). LC/MS(ESI): m/z =459[M+H] ⁺ .

Step B: 7-chloro-2-(1-(pyrrol-1-yl)cyclopropyl)methoxy)-4-(2-oxo-1,3,7-triazaspiro[4.5]decan-7-yl)pyridin[2,3-d]pyrimidine (91.8 mg, 0.2 mmol), 3-methoxy-8-ethyl-7-fluoroisoquinolinylboronic acid pinacol ester (66.4 mg, 0.2 mmol), [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (16 mg, 0.022 mmol), K ₃ PO ₄ , THF (5 mL) and water (1 mL) were mixed, then heated to 60° C. under reflux and stirred for 16 hours. The reaction was cooled to room temperature and stirred overnight to give a light yellow precipitate. The reaction mixture was diluted with water (2 mL) and the solid was collected by filtration. The crude product was chromatographed on a methanol column to give 7-(3-methoxy-8-ethyl-7-fluoroisoquinolin-1-yl)-4-(2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)-2-(1-(pyrrol-1-yl)cyclopropyl)methoxy)pyridin[2,3-d]pyrimidine (74 mg, yield 59%). LC/MS(ESI): m/z =627[M+H] ⁺ .

Step C: 7-(3-methoxy-8-ethyl-7-fluoroisoquinolin-1-yl)-4-(2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)-2-(1-(pyrrol-1-yl)cyclopropyl)methoxy)pyridin[2,3-d]pyrimidine (62.7 mg, 0.1 mmol) was dissolved in DCM (2 mL) at -78°C, BBr ₃ ((0.4 mL, 1M in DCM, 0.4 mmol) was added, and then the mixture was heated to room temperature and stirred for 5 h, then cooled to -78°C, adjusted to Ph = 7 with 1N NaOH, extracted with EA, and then concentrated by filtration under reduced pressure and purified by preparative HPLC to give compound 4 (34 mg, yield 55%). LC/MS(ESI): m/z =613[M+H] ⁺ .

Embodiment 5

Step A: Mix 2,7-dichloro-4-(2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)pyridin[2,3-d]pyrimidine (250 mg, 1 mmol), (2R,8S)-2-fluoro-1,2,3,5,6,7-hexahydropyrrolizin-7-yl]methanol (148 mg, 1.1 mmol), potassium carbonate (0.12 mg, 1.4 mmol), a catalytic amount of potassium iodide and DMF (8 mL), heat to 120°C, and stir for 4 hours. The mixture was cooled to room temperature and evaporated under reduced pressure to give yellow solid 7-chloro-2(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-(2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)pyridin[2,3-d]pyrimidine (0.387 g, 72%). LC/MS(ESI): m/z =538.2[M+H] ⁺ .

Step B: 7-chloro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-(2-oxo-1,3,7-triazaspiro[4.5]dec-7-yl)pyridin[2,3-d]pyrimidine (107 mg, 0.2 mmol), 3-methoxy-8-chloro-7-fluoroisoquinolinylboronic acid pinacol ester (67.6 mg, 0.2 mmol), [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (16 mg, 0.022 mmol), _K3PO4 , THF (5 mL) and water (1 mL) were _added . mL) and then refluxed to 60 ° C and stirred for 16 hours. The reaction was cooled to room temperature and stirred overnight to obtain a light yellow precipitate. The reaction mixture was diluted with water (2 mL) and the solid was collected by filtration. The crude product was column chromatographed to obtain a beige solid 7-(3-methoxy-8-chloro-7-fluoroisoquinolin-1-yl)-4-(2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[2,3-d]pyrimidine (94 mg, yield 58%). LC/MS(ESI): m/z =809.3[M+H] ⁺ .

Step C: 7-(3-methoxy-8-chloro-7-fluoroisoquinolin-1-yl)-4-(2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[2,3-d]pyrimidine (81 mg, 0.1 mmol) was dissolved in DCM (2 mL) at -78°C, BBr ₃ ((0.4 mL, 1M in DCM, 0.4 mmol) was added, and then the mixture was heated to room temperature and stirred for 5 h, then cooled to -78°C and heated with 1N The solution was adjusted to Ph = 7 with NaOH, extracted with EA, and then filtered, concentrated under reduced pressure, and purified by preparative HPLC to obtain a light yellow solid compound 5 (32 mg, yield 56%). LC/MS (ESI): m/z = 579.2 [M + H] ⁺ .

Embodiment 6

Step A: Mix 2,4,7-trichloropyrido[2,3-d]pyrimidine (2.52 g, 10 mmol), 2-thia-1,3,7-triazaspiro[4.5]decane-2,2-one (2.37 g, 11 mmol), potassium carbonate (2.07 g, 15 mmol), catalytic amount of potassium iodide and DMF (60 mL), heat to 120°C, and stir for 4 hours. Cool to room temperature, evaporate under reduced pressure to obtain yellow solid 2,7-dichloro-4-(2-thia-1,3,7-triazaspiro[4.5]decane-2,2-one-7-yl)pyrido[2,3-d]pyrimidine (3.61 g). LC/MS(ESI): m/z =390[M+H] ⁺ .

Step B: Mix 2,7-dichloro-4-2-thia-1,3,7-triazaspiro[4.5]decane-2,2-one-7-yl)pyridin[2,3-d]pyrimidine (390 mg, 1 mmol), (2R,8S)-2-fluoro-1,2,3,5,6,7-hexahydropyrrolizin-7-yl]methanol (175 mg, 1.1 mmol), potassium carbonate (0.18 mg, 2.1 mmol) a catalytic amount of potassium iodide and DMF (8 mL), heat to 120°C, and stir for 4 hours. The mixture was cooled to room temperature and evaporated under reduced pressure to give yellow solid 7-chloro-2(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-(2-thia-1,3,7-triazaspiro[4.5]decane-2,2-one-7-yl)pyridin[2,3-d]pyrimidine (0.417 g). LC/MS(ESI): m/z =513[M+H] ⁺ .

Step C: 7-chloro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-(2-thia-1,3,7-triazaspiro[4.5]decan-2,2-one-7-yl)pyridin[2,3-d]pyrimidine (126 mg, 0.2 mmol), 3-methoxy-8-chloro-7-fluoroisoquinolinylboronic acid pinacol ester (67.6 mg, 0.2 mmol), [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (16 mg, 0.022 mmol), K ₃ PO ₄ , THF (5 mL) and water (1 mL) were added to a flask. mL) and then heated to 60°C under reflux and stirred for 16 hours. The reaction mixture was cooled to room temperature and stirred overnight to obtain a light yellow precipitate. The reaction mixture was diluted with water (2 mL) and the solid was collected by filtration. The crude product was column chromatographed to obtain a beige solid 7-(3-methoxy-8-chloro-7-fluoroisoquinolin-1-yl)-4-(2-thia-1,3,7-triazaspiro[4.5]decane-2,2-one-7-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[2,3-d]pyrimidine (98 mg). LC/MS(ESI): m/z =688[M+H] ⁺ .

Step D: 7-(3-methoxy-8-chloro-7-fluoroisoquinolin-1-yl)-4-(2-thia-1,3,7-triazaspiro[4.5]decan-2,2-one-7-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[2,3-d]pyrimidine (69 mg, 0.1 mmol) was dissolved in DCM (2 mL) at -78°C, BBr ₃ ((0.4 mL, 1M in DCM, 0.4 mmol) was added, and the mixture was heated to room temperature and stirred for 5 h, then cooled to -78°C and heated with 1N The solution was adjusted to Ph = 7 with NaOH, extracted with EA, and then filtered, concentrated under reduced pressure, and purified by preparative HPLC to obtain a light yellow solid compound 6 (29 mg). LC/MS (ESI): m/z = 674 [M + H] ⁺ .

Embodiment 7

Step A: 7-chloro-2(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-(2-oxo-1,3,7-triazaspiro[4.5]dec-7-yl)pyridin[2,3-d]pyrimidine (126 mg, 0.2 mmol), 3-methoxy-8-ethyl-7-fluoroisoquinolinylboronic acid pinacol ester (66.4 mg, 0.2 mmol), [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (16 mg, 0.022 mmol), K ₃ PO ₄ , THF (5 mL) and water (1 mL) were mixed, then heated to 60° C. under reflux and stirred for 16 hours. The reaction was cooled to room temperature and stirred overnight to give a light yellow precipitate. The reaction mixture was diluted with water (2 mL) and the solid was collected by filtration. The crude product was column chromatographed to give a beige solid 7-(3-methoxy-8-ethyl-7-fluoroisoquinolin-1-yl)-4-(2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[2,3-d]pyrimidine (92 mg). LC/MS(ESI): m/z =645[M+H] ⁺ .

Step B: 7-(3-methoxy-8-ethyl-7-fluoroisoquinolin-1-yl)-4-(2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[2,3-d]pyrimidine (64.5 mg, 0.1 mmol) was dissolved in DCM (2 mL) at -78°C, BBr ₃ ((0.4 mL, 1M in DCM, 0.4 mmol) was added, and then the mixture was heated to room temperature and stirred for 5 h, then cooled to -78°C and heated with 1N The solution was adjusted to Ph = 7 with NaOH, extracted with EA, and then filtered, concentrated under reduced pressure, and purified by preparative HPLC to obtain a light yellow solid compound 7 (28 mg). LC/MS (ESI): m/z = 631 [M + H] ⁺ .

Embodiment 8

Step A: 7-chloro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-(2-thia-1,3,7-triazaspiro[4.5]decan-2,2-one-7-yl)pyridin[2,3-d]pyrimidine (102 mg, 0.2 mmol), 3-methoxy-8-ethyl-7-fluoroisoquinolinylboronic acid pinacol ester (66.4 mg, 0.2 mmol), [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (16 mg, 0.022 mmol), K ₃ PO ₄ , THF (5 mL) and water (1 mL) were added to a flask. mL) and then heated to 60°C under reflux and stirred for 16 hours. The reaction mixture was cooled to room temperature and stirred overnight to obtain a light yellow precipitate. The reaction mixture was diluted with water (2 mL) and the solid was collected by filtration. The crude product was column chromatographed to obtain a beige solid 7-(3-methoxy-8-ethyl-7-fluoroisoquinolin-1-yl)-4-(2-thia-1,3,7-triazaspiro[4.5]decane-2,2-one-7-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[2,3-d]pyrimidine (94 mg). LC/MS(ESI): m/z =681[M+H] ⁺ .

Step B: 7-(3-methoxy-8-ethyl-7-fluoroisoquinolin-1-yl)-4-(2-thia-1,3,7-triazaspiro[4.5]decan-2,2-one-7-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[2,3-d]pyrimidine (68 mg, 0.1 mmol) was dissolved in DCM (2 mL) at -78°C, BBr ₃ ((0.4 mL, 1M in DCM, 0.4 mmol) was added, and then the mixture was heated to room temperature and stirred for 5 h, then cooled to -78°C and heated with 1N The solution was adjusted to Ph = 7 with NaOH, extracted with EA, and then concentrated by filtration under reduced pressure and purified by preparative HPLC to obtain a light yellow solid compound 7 (32 mg). LC/MS (ESI): m/z = 667 [M + H] ⁺ .

Embodiment 9

Step A: Mix 2,4,7-trichloropyrido[2,3-d]pyrimidine (2.52 g, 10 mmol), (3R)-3-methylpiperidin-3-ol hydrochloride (1.66 g, 11 mmol), potassium carbonate (2.07 g, 15 mmol), catalytic amount of potassium iodide and DMF (60 mL), heat to 120°C, and stir for 4 hours. Cool to room temperature, evaporate under reduced pressure to obtain yellow solid 2,7-dichloro-4-((3R)-3-methyl-3-hydroxypiperidinyl)pyrido[2,3-d]pyrimidine (2.81 g). LC/MS(ESI): m/z =314[M+H] ⁺ .

Step B: 2,7-Dichloro-4-((3R)-3-methyl-3-hydroxypiperidinyl)pyridin[2,3-d]pyrimidine (314 mg, 1 mmol), (2R,8S)-2-fluoro-1,2,3,5,6,7-hexahydropyrrolizin-7-yl]methanol (175 mg, 1.1 mmol), potassium carbonate (0.18 mg, 2.1 mmol) a catalytic amount of potassium iodide and DMF (8 mL) were mixed, heated to 120°C, and stirred for 4 hours. The mixture was cooled to room temperature and evaporated under reduced pressure to give yellow solid 7-chloro-2(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-((3R)-3-methyl-3-hydroxypiperidinyl)pyridin[2,3-d]pyrimidine (0.289 g). LC/MS(ESI): m/z =436[M+H] ⁺ .

Step C: 7-chloro-2(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-((3R)-3-methyl-3-hydroxypiperidinyl)pyridin[2,3-d]pyrimidine (87 mg, 0.2 mmol), 3-methoxy-8-methylisoquinolinylboronic acid pinacol ester (60 mg, 0.2 mmol), [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (16 mg, 0.022 mmol), K ₃ PO ₄ , THF (5 mL) and water (1 mL) were mixed, then heated to 60° C. under reflux and stirred for 16 hours. The reaction was cooled to room temperature and stirred overnight to give a light yellow precipitate. The reaction mixture was diluted with water (2 mL) and the solid was collected by filtration. The crude product was column chromatographed to give a beige solid 7-(3-methoxy-8-methylisoquinolin-1-yl)-4-((3R)-3-methyl-3-hydroxypiperidinyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[2,3-d]pyrimidine (96 mg). LC/MS(ESI): m/z =573[M+H] ⁺ .

Step D: 7-(3-methoxy-8-methylisoquinolin-1-yl)-4-((3R)-3-methyl-3-hydroxypiperidinyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[2,3-d]pyrimidine (57.3 mg, 0.1 mmol) was dissolved in DCM (2 mL) at -78°C, BBr ₃ ((0.4 mL, 1M in DCM, 0.4 mmol) was added, and then the mixture was heated to room temperature and stirred for 5 h, then cooled to -78°C, adjusted to Ph = 7 with 1N NaOH, extracted with EA, and then concentrated by filtration under reduced pressure and purified by preparative HPLC to give a light yellow solid compound 9 (31 mg). LC/MS(ESI): m/z =559[M+H] ⁺ .

Embodiment 10

Step A: 7-chloro-2(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-((3R)-3-methyl-3-hydroxypiperidinyl)pyridin[2,3-d]pyrimidine (87 mg, 0.2 mmol), 3-methoxy-8-fluoroisoquinolinylboronic acid pinacol ester (60.8 mg, 0.2 mmol), methanesulfonic acid [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) (16 mg, 0.022 mmol), K ₃ PO ₄ , THF (5 mL) and water (1 mL) were mixed, then heated to 60° C. under reflux and stirred for 16 hours. The reaction was cooled to room temperature and stirred overnight to give a light yellow precipitate. The reaction mixture was diluted with water (2 mL) and the solid was collected by filtration. The crude product was column chromatographed to give a beige solid 7-(3-methoxy-8-fluoroisoquinolin-1-yl)-4-((3R)-3-methyl-3-hydroxypiperidinyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[2,3-d]pyrimidine (98 mg). LC/MS(ESI): m/z =577[M+H] ⁺ .

Step B: 7-(3-methoxy-8-fluoroisoquinolin-1-yl)-4-((3R)-3-methyl-3-hydroxypiperidinyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[2,3-d]pyrimidine (57.3 mg, 0.1 mmol) was dissolved in DCM (2 mL) at -78°C, BBr ₃ ((0.4 mL, 1M in DCM, 0.4 mmol) was added, and then the mixture was heated to room temperature and stirred for 5 h, then cooled to -78°C, adjusted to Ph = 7 with 1N NaOH, extracted with EA, and then concentrated by filtration under reduced pressure and purified by preparative HPLC to give a light yellow solid compound 10 (30 mg). LC/MS(ESI): m/z =563[M+H] ⁺ .

Embodiment 11

Step A: 7-chloro-2(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-((3R)-3-methyl-3-hydroxypiperidinyl)pyridin[2,3-d]pyrimidine (87 mg, 0.2 mmol), 3-methoxyisoquinolinylboronic acid pinacol ester (57 mg, 0.2 mmol), methanesulfonic acid [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) (16 mg, 0.022 mmol), K ₃ PO ₄ , THF (5 mL) and water (1 mL) were mixed, then heated to 60° C. under reflux and stirred for 16 hours. The reaction was cooled to room temperature and stirred overnight to give a light yellow precipitate. The reaction mixture was diluted with water (2 mL) and the solid was collected by filtration. The crude product was column chromatographed to give 7-(3-methoxyisoquinolin-1-yl)-4-((3R)-3-methyl-3-hydroxypiperidinyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[2,3-d]pyrimidine (94 mg) as a beige solid. LC/MS(ESI): m/z =559[M+H] ⁺ .

Step B: 7-(3-methoxyisoquinolin-1-yl)-4-((3R)-3-methyl-3-hydroxypiperidinyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[2,3-d]pyrimidine (55.9 mg, 0.1 mmol) was dissolved in DCM (2 mL) at -78°C, BBr ₃ ((0.4 mL, 1M in DCM, 0.4 mmol) was added, and then the mixture was heated to room temperature and stirred for 5 h, then cooled to -78°C, adjusted to Ph = 7 with 1N NaOH, extracted with EA, and then concentrated by filtration under reduced pressure and purified by preparative HPLC to give a light yellow solid compound 11 (32 mg). LC/MS(ESI): m/z =544[M+H] ⁺ .

Embodiment 12

Step A: Mix 2,4,7-trichloro-6-fluoropyrido[2,3-d]pyrimidine (2.52 g, 10 mmol), (3R)-3-methylpiperidin-3-ol hydrochloride (1.66 g, 11 mmol), potassium carbonate (2.07 g, 15 mmol), catalytic amount of potassium iodide and DMF (60 mL), heat to 120°C, and stir for 4 hours. Cool to room temperature, evaporate under reduced pressure to obtain yellow solid 2,7-dichloro-4-((3R)-3-methyl-3-hydroxypiperidinyl)-6-fluoropyrido[2,3-d]pyrimidine (2.88 g). LC/MS(ESI): m/z =332[M+H] ⁺ .

Step B: 2,7-Dichloro-4-((3R)-3-methyl-3-hydroxypiperidinyl)-6-fluoropyrido[2,3-d]pyrimidine (332 mg, 1 mmol), (2R,8S)-2-fluoro-1,2,3,5,6,7-hexahydropyrrolizin-7-yl]methanol (175 mg, 1.1 mmol), potassium carbonate (0.18 mg, 2.1 mmol) a catalytic amount of potassium iodide and DMF (8 mL) were mixed, heated to 120°C, and stirred for 4 hours. The mixture was cooled to room temperature and evaporated under reduced pressure to give yellow solid 7-chloro-6-fluoro-2(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-((3R)-3-methyl-3-hydroxypiperidinyl)pyridin[2,3-d]pyrimidine (0.369 g). LC/MS(ESI): m/z =453[M+H] ⁺ .

Step C: 7-Chloro-6-fluoro-2(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-((3R)-3-methyl-3-hydroxypiperidinyl)pyridin[2,3-d]pyrimidine (87 mg, 0.2 mmol), 3-methoxy-8-chloroisoquinolinylboronic acid pinacol ester (64 mg, 0.2 mmol), [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (16 mg, 0.022 mmol), K ₃ PO ₄ , THF (5 mL) and water (1 mL) were mixed, then heated to 60° C. under reflux and stirred for 16 hours. The reaction was cooled to room temperature and stirred overnight to give a light yellow precipitate. The reaction mixture was diluted with water (2 mL) and the solid was collected by filtration. The crude product was column chromatographed to give a beige solid 6-fluoro-7-(3-methoxy-8-chloroisoquinolin-1-yl)-4-((3R)-3-methyl-3-hydroxypiperidinyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[2,3-d]pyrimidine (92 mg). LC/MS(ESI): m/z =612[M+H] ⁺ .

Step D: At -78°C, 6-fluoro-7-(3-methoxy-8-chloroisoquinolin-1-yl)-4-((3R)-3-methyl-3-hydroxypiperidinyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[2,3-d]pyrimidine (61.2 mg, 0.1 mmol) was dissolved in DCM (2 mL), and BBr ₃ ((0.4 mL, 1M in DCM, 0.4 mmol) was added, and then the mixture was heated to room temperature and stirred for 5 h, then cooled to -78°C, adjusted to Ph = 7 with 1N NaOH, extracted with EA, and then filtered under reduced pressure and concentrated and purified by preparative HPLC to obtain a light yellow solid compound 12. (31 mg). LC/MS(ESI): m/z =598[M+H] ⁺ .

Embodiment 13

Step A: 7-chloro-6-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-((3R)-3-methyl-3-hydroxypiperidinyl)pyridin[2,3-d]pyrimidine (87 mg, 0.2 mmol), 3-methoxy-7-fluoro-8-((triisopropylsilyl)ethynyl)isoquinolinylboronic acid pinacol ester (96.6 mg, 0.2 mmol), [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (16 mg, 0.022 mmol), K ₃ PO ₄ , THF (5 mL) and water (1 mL) were added to a flask. mL) and then refluxed to 60 ° C and stirred for 16 hours. The reaction was cooled to room temperature and stirred overnight to obtain a light yellow precipitate. The reaction mixture was diluted with water (2 mL) and the solid was collected by filtration. The crude product was column chromatographed to obtain a beige solid 6-fluoro-7-(3-methoxy 8-((triisopropylmethylsilyl)ethynyl)isoquinolin-1-yl)-4-((3R)-3-methyl-3-hydroxypiperidinyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[2,3-d]pyrimidine (122 mg). LC/MS(ESI): m/z =776[M+H] ⁺ .

Step B: 6-Fluoro-7-(3-methoxy-8-((triisopropylsilyl)ethynyl)isoquinolin-1-yl)-4-((3R)-3-methyl-3-hydroxypiperidinyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[2,3-d]pyrimidine (77.6 mg, 0.1 mmol) was dissolved in DMF (5 mL) at -78°C. CsF (152 mg, 1 mmol) was added under N2 _- protection. The mixture was stirred at 20°C for 1 hour, then poured into water, extracted with EA, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was dissolved in DCM (2 mL), and BBr ₃ ((0.4 mL, 1M in DCM , 0.4 mmol) was added, then the mixture was heated to room temperature and stirred for 5 h, then cooled to -78°C, adjusted to Ph = 7 with 1N NaOH, extracted with EA, then filtered and concentrated under reduced pressure and purified by preparative HPLC to obtain light yellow solid compound 12 (27 mg). LC/MS (ESI): m/z = 605 [M + H] ⁺ .

Embodiment 14

Step A: 7-chloro-6-fluoro-2(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-((3R)-3-methyl-3-hydroxypiperidinyl)pyridin[2,3-d]pyrimidine (87 mg, 0.2 mmol), 3-methoxy-8-ethyl-7-fluoroisoquinolinylboronic acid pinacol ester (66.2 mg, 0.2 mmol), methanesulfonic acid [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) (16 mg, 0.022 mmol), K ₃ PO ₄ , THF (5 mL) and water (1 mL) were mixed, then heated to 60° C. under reflux and stirred for 16 hours. The reaction was cooled to room temperature and stirred overnight to give a light yellow precipitate. The reaction mixture was diluted with water (2 mL) and the solid was collected by filtration. The crude product was column chromatographed to give 6-fluoro-7-(3-methoxy-8-ethyl-7-fluoroisoquinolin-1-yl)-4-((3R)-3-methyl-3-hydroxypiperidinyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[2,3-d]pyrimidine (94 mg) as a beige solid. LC/MS(ESI): m/z =623[M+H] ⁺ .

Step B: At -78°C, 6-fluoro-7-(3-methoxy-8-ethyl-7-fluoroisoquinolin-1-yl)-4-((3R)-3-methyl-3-hydroxypiperidinyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[2,3-d]pyrimidine (62.3 mg, 0.1 mmol) was dissolved in DCM (2 mL), BBr ₃ ((0.4 mL, 1M in DCM, 0.4 mmol) was added, and then the mixture was heated to room temperature and stirred for 5 h, then cooled to -78°C and heated with 1N The solution was adjusted to Ph = 7 with NaOH, extracted with EA, and then filtered, concentrated under reduced pressure, and purified by preparative HPLC to obtain a light yellow solid compound 14 (32 mg). LC/MS (ESI): m/z = 609 [M + H] ⁺ .

Embodiment 14

Step A: 7-chloro-2(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-((3R)-3-methyl-3-hydroxypiperidinyl)pyridin[2,3-d]pyrimidine (87 mg, 0.2 mmol), 3-methoxy-8-ethyl-7-fluoroisoquinolinylboronic acid pinacol ester (96.3 mg, 0.2 mmol), [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (16 mg, 0.022 mmol), K ₃ PO ₄ , THF (5 mL) and water (1 mL) were mixed, then heated to 60° C. under reflux and stirred for 16 hours. The reaction was cooled to room temperature and stirred overnight to give a light yellow precipitate. The reaction mixture was diluted with water (2 mL) and the solid was collected by filtration. The crude product was column chromatographed to give 7-(3-methoxy-8-ethyl-7-fluoroisoquinolin-1-yl)-4-((3R)-3-methyl-3-hydroxypiperidinyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[2,3-d]pyrimidine (112 mg) as a beige solid. LC/MS(ESI): m/z =758[M+H] ⁺ .

Step B: 7-(3-methoxy-8-ethyl-7-fluoroisoquinolin-1-yl)-4-((3R)-3-methyl-3-hydroxypiperidinyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[2,3-d]pyrimidine (75.8 mg, 0.1 mmol) was dissolved in DMF (5 mL) at -78°C. CsF (152 mg, 1 mmol) was added under N2 _- protection. The mixture was stirred at 20°C for 1 hour, then poured into water, extracted with EA, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was dissolved in DCM (2 mL), and BBr ₃ ((0.4 mL, 1M in DCM , 0.4 mmol) was added, then the mixture was heated to room temperature and stirred for 5 h, then cooled to -78°C, adjusted to Ph = 7 with 1N NaOH, extracted with EA, then filtered and concentrated under reduced pressure and purified by preparative HPLC to obtain light yellow solid compound 15 (24 mg). LC/MS (ESI): m/z = 587 [M + H] ⁺ .

Embodiment 16

Step A: 7-chloro-2(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-((3R)-3-methyl-3-hydroxypiperidinyl)pyridin[2,3-d]pyrimidine (87 mg, 0.2 mmol), 3-methoxy-8-ethyl-7-fluoroisoquinolinylboronic acid pinacol ester (66.2 mg, 0.2 mmol), methanesulfonic acid [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) (16 mg, 0.022 mmol), K ₃ PO ₄ , THF (5 mL) and water (1 mL) were mixed, then heated to 60° C. under reflux and stirred for 16 hours. The reaction was cooled to room temperature and stirred overnight to give a light yellow precipitate. The reaction mixture was diluted with water (2 mL) and the solid was collected by filtration. The crude product was column chromatographed to give a beige solid 7-(3-methoxy-8-ethyl-7-fluoroisoquinolin-1-yl)-4-((3R)-3-methyl-3-hydroxypiperidinyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[2,3-d]pyrimidine (91 mg). LC/MS(ESI): m/z =605[M+H] ⁺ .

Step B: 7-(3-methoxy-8-ethyl-7-fluoroisoquinolin-1-yl)-4-((3R)-3-methyl-3-hydroxypiperidinyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[2,3-d]pyrimidine (60.4 mg, 0.1 mmol) was dissolved in DCM (2 mL) at -78°C, BBr ₃ ((0.4 mL, 1M in DCM, 0.4 mmol) was added, and then the mixture was heated to room temperature and stirred for 5 h, then cooled to -78°C, adjusted to Ph = 7 with 1N NaOH, extracted with EA, and then filtered under reduced pressure and concentrated and purified by preparative HPLC to obtain a light yellow solid compound 16. (34 mg). LC/MS(ESI): m/z =591[M+H] ⁺ .

Embodiment 17

Step A: Mix 2,4,7-trichloro-8-fluoropyrido[4,3-d]pyrimidine (2.52 g, 10 mmol), 8-boc-3,8-diazabicyclo[3.2.1]octane (2.33 g, 11 mmol), potassium carbonate (2.07 g, 15 mmol), catalytic amount of potassium iodide and DMF (60 mL), heat to 120°C, and stir for 4 hours. Cool to room temperature, evaporate under reduced pressure to obtain yellow solid 2,7-dichloro-4-((3R)-3-methyl-3-hydroxypiperidinyl)-8-fluoropyrido[4,3-d]pyrimidine (3.71 g). LC/MS(ESI): m/z =428[M+H] ⁺ .

Step B: 2,7-Dichloro-4-((3R)-3-methyl-3-hydroxypiperidinyl)pyridin[2,3-d]pyrimidine (428 mg, 1 mmol), (2R,8S)-2-fluoro-1,2,3,5,6,7-hexahydropyrrolizin-7-yl]methanol (175 mg, 1.1 mmol), potassium carbonate (0.18 mg, 2.1 mmol) a catalytic amount of potassium iodide and DMF (8 mL) were mixed, heated to 120°C, and stirred for 4 hours. The mixture was cooled to room temperature and evaporated under reduced pressure to give yellow solid 7-chloro-2(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-((3R)-3-methyl-3-hydroxypiperidinyl)-8-fluoropyrido[4,3-d]pyrimidine (469 mg). LC/MS(ESI): m/z =552[M+H] ⁺ .

Step C: 7-chloro-2(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-((3R)-3-methyl-3-hydroxypiperidinyl)-8-fluoropyrido[4,3-d]pyrimidine (110.4 mg, 0.2 mmol), 3-methoxy-8-ethylisoquinolinylboronic acid pinacol ester (66.4 mg, 0.2 mmol), [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (16 mg, 0.022 mmol), K ₃ PO ₄ , THF (5 mL) and water (1 mL) were mixed, then heated to 60° C. under reflux and stirred for 16 hours. The reaction was cooled to room temperature and stirred overnight to give a light yellow precipitate. The reaction mixture was diluted with water (2 mL) and the solid was collected by filtration. The crude product was column chromatographed to give 7-(3-methoxy-8-ethylisoquinolin-1-yl)-4-((3R)-3-methyl-3-hydroxypiperidinyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-8-fluoropyrido[4,3-d]pyrimidine (98 mg) as a beige solid. LC/MS(ESI): m/z =720[M+H] ⁺ .

Step D: 7-(3-methoxy-8-methylisoquinolin-1-yl)-4-((3R)-3-methyl-3-hydroxypiperidinyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-8-fluoropyrido[4,3-d]pyrimidine (72.0 mg, 0.1 mmol) was dissolved in DCM (2 mL) at -78°C, BBr ₃ ((0.4 mL, 1M in DCM, 0.4 mmol) was added, and then the mixture was heated to room temperature and stirred for 5 h, then cooled to -78°C, adjusted to Ph = 7 with 1N NaOH, extracted with EA, and then filtered under reduced pressure and concentrated and purified by preparative HPLC to obtain a light yellow solid compound 17. (29 mg). LC/MS(ESI): m/z =606[M+H] ⁺ .

Embodiment 18 Compound 18 (28 mg) was obtained by a method similar to Example 17. LC/MS (ESI): m/z = 613 [M+H] ⁺ .

Embodiment 19

Step A: Mix 2,4,7-trichloro-8-fluoropyrido[4,3-d]pyrimidine (2.52 g, 10 mmol), (3R)-3-methylpiperidin-3-ol hydrochloride (1.66 g, 11 mmol), potassium carbonate (2.07 g, 15 mmol), catalytic amount of potassium iodide and DMF (60 mL), heat to 120°C, and stir for 4 hours. Cool to room temperature, evaporate under reduced pressure to obtain yellow solid 2,7-dichloro-4-((3R)-3-methyl-3-hydroxypiperidinyl)-8-fluoropyrido[4,3-d]pyrimidine (2.82 g). LC/MS(ESI): m/z =332[M+H] ⁺ .

Step B: 2,7-Dichloro-4-((3R)-3-methyl-3-hydroxypiperidinyl)-8-fluoropyrido[4,3-d]pyrimidine (332 mg, 1 mmol), (2R,8S)-2-fluoro-1,2,3,5,6,7-hexahydropyrrolizin-7-yl]methanol (175 mg, 1.1 mmol), potassium carbonate (0.18 mg, 2.1 mmol), a catalytic amount of potassium iodide and DMF (8 mL) were mixed, heated to 120°C, and stirred for 4 hours. The mixture was cooled to room temperature and evaporated under reduced pressure to give yellow solid 7-chloro-8-fluoro-2(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-((3R)-3-methyl-3-hydroxypiperidinyl)pyrido[4,3-d]pyrimidine (0.368 g). LC/MS(ESI): m/z =453[M+H] ⁺ .

Step C: 7-chloro-8-fluoro-2(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-((3R)-3-methyl-3-hydroxypiperidinyl)pyridin[4,3-d]pyrimidine (87 mg, 0.2 mmol), 3-methoxy-8-ethylisoquinolinylboronic acid pinacol ester (66.4 mg, 0.2 mmol), methanesulfonic acid [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) (16 mg, 0.022 mmol), K ₃ PO ₄ , THF (5 mL) and water (1 mL) were mixed, then heated to 60° C. under reflux and stirred for 16 hours. The reaction was cooled to room temperature and stirred overnight to give a light yellow precipitate. The reaction mixture was diluted with water (2 mL) and the solid was collected by filtration. The crude product was column chromatographed to give 8-fluoro-7-(3-methoxy-8-ethylisoquinolin-1-yl)-4-((3R)-3-methyl-3-hydroxypiperidinyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[4,3-d]pyrimidine (92 mg) as a beige solid. LC/MS(ESI): m/z =623[M+H] ⁺ .

Step D: 8-Fluoro-7-(3-methoxy-8-ethylisoquinolin-1-yl)-4-((3R)-3-methyl-3-hydroxypiperidinyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[4,3-d]pyrimidine (61.2 mg, 0.1 mmol) was dissolved in DCM (2 mL) at -78°C, BBr ₃ ((0.4 mL, 1M in DCM, 0.4 mmol) was added, and then the mixture was heated to room temperature and stirred for 5 h, then cooled to -78°C, adjusted to Ph = 7 with 1N NaOH, extracted with EA, and then filtered under reduced pressure and concentrated and purified by preparative HPLC to obtain a light yellow solid compound 19. (30 mg). LC/MS(ESI): m/z =609[M+H] ⁺ .

Embodiment 20

Step A: Mix 2,4,7-trichloro-8-fluoropyrido[4,3-d]pyrimidine (2.52 g, 10 mmol), (3S)-3-methylpiperidin-3-ol hydrochloride (1.66 g, 11 mmol), potassium carbonate (2.07 g, 15 mmol), catalytic amount of potassium iodide and DMF (60 mL), heat to 120°C, and stir for 4 hours. Cool to room temperature, evaporate under reduced pressure to obtain yellow solid 2,7-dichloro-4-((3S)-3-methyl-3-hydroxypiperidinyl)-8-fluoropyrido[4,3-d]pyrimidine (2.88 g). LC/MS(ESI): m/z =332[M+H] ⁺ .

Step B: 2,7-Dichloro-4-((3S)-3-methyl-3-hydroxypiperidinyl)-8-fluoropyrido[4,3-d]pyrimidine (332 mg, 1 mmol), (2R,8S)-2-fluoro-1,2,3,5,6,7-hexahydropyrrolizin-7-yl]methanol (175 mg, 1.1 mmol), potassium carbonate (0.18 mg, 2.1 mmol), a catalytic amount of potassium iodide and DMF (8 mL) were mixed, heated to 120°C, and stirred for 4 hours. The mixture was cooled to room temperature and evaporated under reduced pressure to give yellow solid 7-chloro-8-fluoro-2(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-((3S)-3-methyl-3-hydroxypiperidinyl)pyrido[4,3-d]pyrimidine (0.378 g). LC/MS(ESI): m/z =453[M+H] ⁺ .

Step C: 7-chloro-8-fluoro-2(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-((3S)-3-methyl-3-hydroxypiperidinyl)pyridin[4,3-d]pyrimidine (87 mg, 0.2 mmol), 3-methoxy-8-ethylisoquinolinylboronic acid pinacol ester (66.4 mg, 0.2 mmol), [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (16 mg, 0.022 mmol), K ₃ PO ₄ , THF (5 mL) and water (1 mL) were mixed, then heated to 60° C. under reflux and stirred for 16 hours. The reaction was cooled to room temperature and stirred overnight to give a light yellow precipitate. The reaction mixture was diluted with water (2 mL) and the solid was collected by filtration. The crude product was column chromatographed to give 8-fluoro-7-(3-methoxy-8-ethylisoquinolin-1-yl)-4-((3S)-3-methyl-3-hydroxypiperidinyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[4,3-d]pyrimidine (90 mg) as a beige solid. LC/MS(ESI): m/z =623[M+H] ⁺ .

Step D: 8-Fluoro-7-(3-methoxy-8-ethylisoquinolin-1-yl)-4-((3S)-3-methyl-3-hydroxypiperidinyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[4,3-d]pyrimidine (61.2 mg, 0.1 mmol) was dissolved in DCM (2 mL) at -78°C, BBr ₃ ((0.4 mL, 1M in DCM, 0.4 mmol) was added, and then the mixture was heated to room temperature and stirred for 5 h, then cooled to -78°C, adjusted to Ph = 7 with 1N NaOH, extracted with EA, and then filtered under reduced pressure and concentrated and purified by preparative HPLC to obtain a light yellow solid compound 20. (26 mg). LC/MS(ESI): m/z =609[M+H] ⁺ .

Embodiment 21 Compound 21 (28 mg) was obtained by a method similar to Example 19. LC/MS (ESI): m/z = 605 [M+H] ⁺ .

Embodiment 22 Compound 22 (29 mg) was obtained by a method similar to Example 20. LC/MS (ESI): m/z = 605 [M+H] ⁺ .

Embodiment 23 Compound 23 (32 mg) was obtained by a method similar to Example 19. LC/MS (ESI): m/z = 609 [M+H] ⁺ .

Embodiment 24 Compound 24 (28 mg) was obtained by a method similar to Example 20. LC/MS (ESI): m/z = 605 [M+H] ⁺ .

Embodiment 25 Compound 25 (29 mg) was obtained by a method similar to Example 19. LC/MS (ESI): m/z = 604 [M+H] ⁺ .

Embodiment 26 Compound 26 (29 mg) was obtained by a method similar to Example 20. LC/MS (ESI): m/z = 604 [M+H] ⁺ .

Embodiment 27 Compound 27 (24 mg) was obtained by a method similar to Example 20. LC/MS (ESI): m/z = 615 [M+H] ⁺ .

Embodiment 28 Compound 28 (31 mg) was obtained by a method similar to Example 19. LC/MS (ESI): m/z = 615 [M+H] ⁺ .

Embodiment 29 Compound 29 (32 mg) was obtained by a method similar to Example 17. LC/MS (ESI): m/z = 605 [M+H] ⁺ .

Embodiment 30 Compound 30 (27 mg) was obtained by a method similar to Example 18. LC/MS (ESI): m/z = 612 [M+H] ⁺ .

Embodiment 31 Compound 31 (38 mg) was obtained by a method similar to Example 18. LC/MS (ESI): m/z = 608 [M+H] ⁺ .

Embodiment 32 Compound 32 (31 mg) was obtained by a method similar to Example 20. LC/MS (ESI): m/z = 608 [M+H] ⁺ .

Embodiment 33 Compound 33 (24 mg) was obtained by a method similar to Example 14. LC/MS (ESI): m/z = 616 [M+H] ⁺ .

Embodiment 34 Compound 34 (31 mg) was obtained by a method similar to Example 9. LC/MS (ESI): m/z = 598 [M+H] ⁺ .

Embodiment 35

Step A: 7-bromo-2(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-((3R)-3-methyl-3-hydroxypiperidinyl)-6, 8-difluoroquinazoline (100.2 mg, 0.2 mmol), 3-methoxy-8-ethylisoquinolinylboronic acid pinacol ester (67.4 mg, 0.2 mmol), [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (16 mg, 0.022 mmol), K ₃ PO ₄ , THF (5 mL) and water (1 mL) were mixed, then heated to 60° C. under reflux and stirred for 16 hours. The reaction was cooled to room temperature and stirred overnight to give a light yellow precipitate. The reaction mixture was diluted with water (2 mL) and the solid was collected by filtration. The crude product was column chromatographed to give a beige solid 7-(3-methoxy-8-ethylisoquinolin-1-yl)-4-((3R)-3-methyl-3-hydroxypiperidinyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrinyl-7a(5H)-yl)methoxy)-6, 8-difluoroquinazoline (92 mg). LC/MS(ESI): m/z =647[M+H] ⁺ .

Step B: 7-(3-methoxy-8-methylisoquinolin-1-yl)-4-((3R)-3-methyl-3-hydroxypiperidinyl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrinyl-7a(5H)-yl)methoxy)-6, 8-difluoroquinazoline (64.7 mg, 0.1 mmol) was dissolved in DCM (2 mL) at -78°C, BBr ₃ ((0.4 mL, 1M in DCM, 0.4 mmol) was added, and then the mixture was heated to room temperature and stirred for 5 h, then cooled to -78°C, adjusted to Ph = 7 with 1N NaOH, extracted with EA, and then filtered, concentrated under reduced pressure and purified by preparative HPLC to obtain a light yellow solid compound 17 (29 mg). Compound 35 (34 mg) was obtained. LC/MS (ESI): m/z = 633 [M+H] ⁺ .

Embodiment 36 Compound 36 (27 mg) was obtained by a method similar to Example 36. LC/MS (ESI): m/z = 649 [M+H] ⁺ .

Embodiment 37 Compound 37 (29 mg) was obtained by a method similar to Example 36. LC/MS (ESI): m/z = 626 [M+H] ⁺ .

Embodiment 38 Compound 38 (28 mg) was obtained by a method similar to Example 36. LC/MS (ESI): m/z = 643 [M+H] ⁺ .

Embodiment 39 Compound 39 (28 mg) was obtained by a method similar to Example 36. LC/MS (ESI): m/z = 622 [M+H] ⁺ .

Embodiment 40 Compound 40 (32 mg) was obtained by a method similar to Example 36. LC/MS (ESI): m/z = 639 [M+H] ⁺ .

Embodiment 41

Step A: 2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrinylcyclo-7a(5H)-yl)methoxy)-4-(8-boc-3,8-diazabicyclo[3.2.1]octan-3-yl)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidine (100 mg, 0.2 mmol), 1-trifluoromethanesulfonyl-7-fluoro-3-methoxy-8-((triisopropylsilyl)ethynyl)isoquinoline (100.8 mg, 0.2 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (16 mg, 0.022 mmol), Cs ₂ CO ₃ , dioxane (5 mL) and water (1 mL) were added to a molten-crystal. mL) and then heated to 60 ° C under reflux and stirred for 16 hours. The reaction was cooled to room temperature and stirred overnight to obtain a light yellow precipitate. The reaction mixture was diluted with water (2 mL) and the solid was collected by filtration. The crude product was column chromatographed to obtain 7-(3-methoxy 8-((triisopropylmethylsilyl)ethynyl)isoquinolin-1-yl)-4-(8-boc-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-5,6,7,8-tetrahydropyrido[3,4-d]pyrimidine (132 mg). LC/MS(ESI): m/z =845[M+H] ⁺ .

Step B: 7-(3-methoxy-8-((triisopropylsilyl)ethynyl)isoquinolin-1-yl)-4-(8-boc-3,8-diazabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)5,6,7,8-tetrahydropyrido[3,4-d]pyrimidine (126.6 mg, 0.15 mmol) was dissolved in DMF (5 mL) at _-78 °C, and CsF (152 mg, 1 mmol), stirred at 20°C for 1 hour, then poured into water, extracted with EA, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was dissolved in DCM (2 mL), BBr ₃ ((0.4 mL, 1M in DCM , 0.4 mmol) was added, then warmed to room temperature and stirred for 5 hours, then cooled to -78°C, adjusted to Ph = 7 with 1N NaOH, extracted with EA, then filtered and concentrated under reduced pressure and purified by preparative HPLC to obtain a light yellow solid compound 41 (48 mg). LC/MS(ESI): m/z =605[M+H] ⁺ .

Embodiment 42 Compound 42 (43 mg) was obtained in a similar manner to Example 41. LC/MS (ESI): m/z = 591 [M+H] ⁺ .

Embodiment 43 Compound 43 (41 mg) was obtained in a similar manner to Example 41. LC/MS (ESI): m/z = 602 [M+H] ⁺ .

Embodiment 44 Compound 44 (47 mg) was obtained in a similar manner to Example 41. LC/MS (ESI): m/z = 595 [M+H] ⁺ .

Embodiment 45

Step A: Mix 2,4,7-trichloro-6-fluoropyrido[2,3-d]pyrimidine (2.52 g, 10 mmol), 1,3,7-triazaspiro[4.5]decan-2-one (1.69 g, 11 mmol), potassium carbonate (2.07 g, 15 mmol), catalytic amount of potassium iodide and DMF (60 mL), heat to 120°C, and stir for 4 hours. Cool to room temperature, evaporate under reduced pressure to obtain yellow solid 2,7-dichloro-4-((2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)-6-fluoropyrido[2,3-d]pyrimidine (3.41 g). LC/MS(ESI): m/z =372[M+H] ⁺ .

Step B: 2,7-Dichloro-4-((2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)-6-fluoropyrido[2,3-d]pyrimidine (371 mg, 1 mmol), (2R,8S)-2-fluoro-1,2,3,5,6,7-hexahydropyrrolizin-7-yl]methanol (175 mg, 1.1 mmol), potassium carbonate (0.18 mg, 2.1 mmol) and a catalytic amount of potassium iodide and DMF (8 mL) and heated to 120°C, stirred and reacted for 4 hours. The mixture was cooled to room temperature and evaporated under reduced pressure to obtain yellow solid 7-chloro-2(((2R,7aS)-2-fluorotetrahydro-1H-pyridine-7a(5H)-yl)methoxy)-4-((2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)-6-fluoropyrido[2,3-d]pyrimidine (0.387 g). LC/MS(ESI): m/z =493[M+H] ⁺ .

Step C: 7-chloro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-(2-thia-1,3,7-triazaspiro[4.5]decan-2,2-one-7-yl)-6-fluoropyrido[2,3-d]pyrimidine (98 mg, 0.2 mmol), 3-methoxy-8-chloro-7-fluoroisoquinolinylboronic acid pinacol ester (67.6 mg, 0.2 mmol), [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (16 mg, 0.022 mmol), K ₃ PO ₄ , THF (5 mL) and water (1 mL) were added to a flask. mL) and then refluxed to 60°C and stirred for 16 hours. The reaction mixture was cooled to room temperature and stirred overnight to obtain a light yellow precipitate. The reaction mixture was diluted with water (2 mL) and the solid was collected by filtration. The crude product was column chromatographed to obtain a beige solid 7-(3-methoxy-8-chloro-7-fluoroisoquinolin-1-yl)-4-((2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-6-fluoropyridin[2,3-d]pyrimidine (92 mg). LC/MS(ESI): m/z =670[M+H] ⁺ .

Step D: 7-(3-methoxy-8-chloro-7-fluoroisoquinolin-1-yl)-4-((2-oxo-1,3,7-triazaspiro[4.5]decane-7-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-6-fluoropyrido[2,3-d]pyrimidine (67 mg, 0.1 mmol) was dissolved in DCM (2 mL) at -78°C, BBr ₃ ((0.4 mL, 1M in DCM, 0.4 mmol) was added, and then the mixture was heated to room temperature and stirred for 5 h, then cooled to -78°C and heated with 1N The pH was adjusted to 7 with NaOH, extracted with EA, and then concentrated by filtration under reduced pressure and purified by preparative HPLC to obtain a light yellow solid compound 45 (23 mg). LC/MS (ESI): m/z = 656 [M+H] ⁺ .

Embodiment 46 Compound 46 (32 mg) was obtained in a similar manner to Example 45. LC/MS (ESI): m/z = 692 [M+H] ⁺ .

Embodiment 47 Compound 47 (25 mg) was obtained in a similar manner to Example 45. LC/MS (ESI): m/z = 685 [M+H] ⁺ .

Embodiment 48 Compound 48 (22 mg) was obtained in a similar manner to Example 45. LC/MS (ESI): m/z = 683 [M+H] ⁺ .

Embodiment 49 Compound 49 (21 mg) was obtained in a similar manner to Example 45. LC/MS (ESI): m/z = 702 [M+H] ⁺ .

Embodiment 50 Compound 50 (16 mg) was obtained in a similar manner to Example 45. LC/MS (ESI): m/z = 715 [M+H] ⁺ .

Embodiment 51 Compound 51 (22 mg) was obtained in a similar manner to Example 32. LC/MS (ESI): m/z = 620 [M+H] ⁺ .

Embodiment 52 Compound 52 (21 mg) was obtained in a similar manner to Example 32. LC/MS (ESI): m/z = 620 [M+H] ⁺ .

Embodiment 53 Compound 53 (21 mg) was obtained in a similar manner to Example 32. LC/MS (ESI): m/z = 619 [M+H] ⁺ .

Embodiment 54 Compound 54 (18 mg) was obtained in a similar manner to Example 32. LC/MS (ESI): m/z = 637 [M+H] ⁺ .

Embodiment 55 Compound 55 (16 mg) was obtained in a similar manner to Example 32. LC/MS (ESI): m/z = 640 [M+H] ⁺ .

Embodiment 56 Compound 56 (27 mg) was obtained in a similar manner to Example 32. LC/MS (ESI): m/z = 623 [M+H] ⁺ .

Embodiment 57 Compound 57 (29 mg) was obtained in a similar manner to Example 32. LC/MS (ESI): m/z = 623 [M+H] ⁺ .

Embodiment 58

Step A: 2,4,7-trichloro-8-fluoropyrido[4,3-d]pyrimidine (2.52 g, 10 mmol), (5R)-2-methyl-1,3,7-triazaspiro-2-phosphaspiro[4.5]decane-2-one (2.08 g, 11 mmol), potassium carbonate (2.07 g, 15 mmol), catalytic amount of potassium iodide and DMF (60 mL) were mixed, heated to 120°C, and stirred for 4 hours. The mixture was cooled to room temperature and evaporated under reduced pressure to obtain yellow solid 2,7-dichloro-4-((5R)-2-methyl-2-oxo-1,3,7-triazaspiro-2-phosphaspiro[4.5]decane-7-yl)-8-fluoropyrido[4,3-d]pyrimidine (3.42 g). LC/MS(ESI): m/z =406[M+H] ⁺ .

Step B: 2,7-Dichloro-4-((5R)-2-methyl-2-oxo-1,3,7-triazaspiro-2-phosphaspiro[4.5]decane-7-yl)-8-fluoropyrido[4,3-d]pyrimidine (406 mg, 1 mmol), (2R,8S)-2-fluoro-1,2,3,5,6,7-hexahydropyrrolizin-7-yl]methanol (175 mg, 1.1 mmol), potassium carbonate (0.18 mg, 2.1 mmol) a catalytic amount of potassium iodide and DMF (8 mL) were mixed, heated to 120°C, and stirred for 4 hours. The mixture was cooled to room temperature and evaporated under reduced pressure to give yellow solid 7-chloro-8-fluoro-2(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-((5R)-2-methyl-2-oxo-1,3,7-triazaspiro-2-phosphaspiro[4.5]decane-7-yl)pyrido[4,3-d]pyrimidine (387 mg). LC/MS(ESI): m/z =528[M+H] ⁺ .

Step C: 7-chloro-8-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-((5R)-2-methyl-2-oxo-1,3,7-triazaspiro-2-phosphaspiro[4.5]dec-7-yl)pyridin[4,3-d]pyrimidine (105.4 mg, 0.2 mmol), 3-methoxy-7-fluoro-8-ethylisoquinolinylboronic acid pinacol ester (66.4 mg, 0.2 mmol), [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (16 mg, 0.022 mmol), K ₃ PO ₄ , THF (5 mL) and water (1 mL) were added. mL) and then heated to 60°C under reflux and stirred for 16 hours. The reaction mixture was cooled to room temperature and stirred overnight to obtain a light yellow precipitate. The reaction mixture was diluted with water (2 mL) and the solid was collected by filtration. The crude product was column chromatographed to obtain 8-fluoro-7-(3-methoxy-7-fluoro-8-ethylisoquinolin-1-yl)-4-((5R)-2-methyl-2-oxo-1,3,7-triazaspiro-2-phosphaspiro[4.5]decane-7-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[4,3-d]pyrimidine (91 mg). LC/MS(ESI): m/z =697[M+H] ⁺ .

Step D: 8-Fluoro-7-(3-methoxy-7-fluoro-8-ethylisoquinolin-1-yl)-4-((5R)-2-methyl-2-oxo-1,3,7-triazaspiro-2-phosphaspiro[4.5]decan-7-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[4,3-d]pyrimidine (69.7 mg, 0.1 mmol) was dissolved in DCM (2 mL) at -78°C, BBr ₃ ((0.4 mL, 1M in DCM, 0.4 mmol) was added, and the mixture was heated to room temperature and stirred for 5 h, then cooled to -78°C and heated with 1N The solution was adjusted to Ph = 7 with NaOH, extracted with EA, and then concentrated by filtration under reduced pressure and purified by preparative HPLC to obtain a light yellow solid compound 58 (28 mg). LC/MS (ESI): m/z = 683 [M + H] ⁺ .

Embodiment 59 Compound 59 (24 mg) was obtained in a similar manner to Example 58. LC/MS (ESI): m/z = 690 [M+H] ⁺ .

Embodiment 60 Compound 60 (22 mg) was obtained in a similar manner to Example 58. LC/MS (ESI): m/z = 697 [M+H] ⁺ .

Embodiment 61 Compound 61 (20 mg) was obtained in a similar manner to Example 58. LC/MS (ESI): m/z = 695 [M+H] ⁺ .

Embodiment 62

Step A: 7-bromo-2,4-dichloro-6,8-difluoroquinazoline (3.13 g, 10 mmol), (5R)-2-methyl-1,3,7-triazaspiro-2-phosphaspiro[4.5]decane-2-one (2.08 g, 11 mmol), potassium carbonate (2.07 g, 15 mmol), catalytic amount of potassium iodide and DMF (60 mL) were mixed, heated to 120°C, and stirred for 4 hours. The mixture was cooled to room temperature and evaporated under reduced pressure to obtain yellow solid 7-bromo-2-chloro-4-((5R)-2-methyl-2-oxo-1,3,7-triazaspiro-2-phosphaspiro[4.5]decane-7-yl)-6,8-difluoroquinazoline (3.52 g). LC/MS(ESI): m/z =467[M+H] ⁺ .

Step B: 7-Bromo-2-chloro-4-((5R)-2-methyl-2-oxo-1,3,7-triazaspiro-2-phosphaspiro[4.5]decane-7-yl)-6,8-difluoroquinazoline (467 mg, 1 mmol), (2R,8S)-2-fluoro-1,2,3,5,6,7-hexahydropyrrolizin-7-yl]methanol (175 mg, 1.1 mmol), potassium carbonate (0.18 mg, 2.1 mmol) a catalytic amount of potassium iodide and DMF (8 mL) were mixed, heated to 120°C, and stirred for 4 hours. The mixture was cooled to room temperature and evaporated under reduced pressure to give yellow solid 7-bromo-6,8-difluoro-2(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-((5R)-2-methyl-2-oxo-1,3,7-triazaspiro-2-phosphaspiro[4.5]decane-7-yl)quinazoline (384 mg). LC/MS(ESI): m/z =590[M+H] ⁺ .

Step C: 7-Bromo-6,8-difluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-((5R)-2-methyl-2-oxo-1,3,7-triazaspiro-2-phosphaspiro[4.5]dec-7-yl)quinazoline (118 mg, 0.2 mmol), 3-methoxy-7-fluoro-8-ethylisoquinolinylboronic acid pinacol ester (66.4 mg, 0.2 mmol), [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (16 mg, 0.022 mmol), K ₃ PO ₄ , THF (5 mL) and water (1 mL) were added to a flask. mL) and then heated to 60°C under reflux and stirred for 16 hours. The reaction mixture was cooled to room temperature and stirred overnight to obtain a light yellow precipitate. The reaction mixture was diluted with water (2 mL) and the solid was collected by filtration. The crude product was column chromatographed to obtain 6,8-difluoro-7-(3-methoxy-7-fluoro-8-ethylisoquinolin-1-yl)-4-((5R)-2-methyl-2-oxo-1,3,7-triazaspiro-2-phosphaspiro[4.5]decane-7-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)quinazoline (78 mg). LC/MS(ESI): m/z =714[M+H] ⁺ .

Step D: 6,8-difluoro-7-(3-methoxy-7-fluoro-8-ethylisoquinolin-1-yl)-4-((5R)-2-methyl-2-oxo-1,3,7-triazaspiro-2-phosphaspiro[4.5]decane-7-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)quinazoline (71.4 mg, 0.1 mmol) was dissolved in DCM (2 mL) at -78°C, BBr ₃ ((0.4 mL, 1M in DCM, 0.4 mmol) was added, and the mixture was heated to room temperature and stirred for 5 h, then cooled to -78°C and heated with 1N The pH was adjusted to 7 with NaOH, extracted with EA, and then concentrated by filtration under reduced pressure and purified by preparative HPLC to obtain a light yellow solid compound 62 (35 mg). LC/MS (ESI): m/z =700[M+H] ⁺ .

Embodiment 63 Compound 63 (24 mg) was obtained in a similar manner to Example 62. LC/MS (ESI): m/z = 707 [M+H] ⁺ .

Embodiment 64 Compound 64 (14 mg) was obtained in a similar manner to Example 62. LC/MS (ESI): m/z = 714 [M+H] ⁺ .

Embodiment 65 Compound 65 (12 mg) was obtained in a similar manner to Example 62. LC/MS (ESI): m/z = 712 [M+H] ⁺ .

Embodiment 66

Step A: 2,4,7-trichloro-8-fluoropyrido[4,3-d]pyrimidine (2.52 g, 10 mmol), (5R)-2-methyl-1,3,7-triazaspiro-2-phosphaspiro[4.5]decane-2-one (2.08 g, 11 mmol), potassium carbonate (2.07 g, 15 mmol), catalytic amount of potassium iodide and DMF (60 mL) were mixed, heated to 120°C, and stirred for 4 hours. The mixture was cooled to room temperature and evaporated under reduced pressure to obtain yellow solid 2,7-dichloro-4-((5R)-2-methyl-2-oxo-1,3,7-triazaspiro-2-phosphaspiro[4.5]decane-7-yl)-6-fluoropyrido[4,3-d]pyrimidine (3.42 g). LC/MS(ESI): m/z =406[M+H] ⁺ .

Step B: 2,7-Dichloro-4-((5R)-2-methyl-2-oxo-1,3,7-triazaspiro-2-phosphaspiro[4.5]decane-7-yl)-6-fluoropyrido[4,3-d]pyrimidine (406 mg, 1 mmol), (2R,8S)-2-fluoro-1,2,3,5,6,7-hexahydropyrrolizin-7-yl]methanol (175 mg, 1.1 mmol), potassium carbonate (0.18 mg, 2.1 mmol) a catalytic amount of potassium iodide and DMF (8 mL) were mixed, heated to 120°C, and stirred for 4 hours. The mixture was cooled to room temperature and evaporated under reduced pressure to give yellow solid 7-chloro-6-fluoro-2(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-((5R)-2-methyl-2-oxo-1,3,7-triazaspiro-2-phosphaspiro[4.5]decane-7-yl)pyrido[4,3-d]pyrimidine (387 mg). LC/MS(ESI): m/z =528[M+H] ⁺ .

Step C: 7-chloro-6-fluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-((5R)-2-methyl-2-oxo-1,3,7-triazaspiro-2-phosphaspiro[4.5]dec-7-yl)pyridin[4,3-d]pyrimidine (105.4 mg, 0.2 mmol), 3-methoxy-7-fluoro-8-ethylisoquinolinylboronic acid pinacol ester (66.4 mg, 0.2 mmol), [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (16 mg, 0.022 mmol), K ₃ PO ₄ , THF (5 mL) and water (1 mL) were added. mL) and then heated to 60°C under reflux and stirred for 16 hours. The reaction mixture was cooled to room temperature and stirred overnight to obtain a light yellow precipitate. The reaction mixture was diluted with water (2 mL) and the solid was collected by filtration. The crude product was column chromatographed to obtain 6-fluoro-7-(3-methoxy-7-fluoro-8-ethylisoquinolin-1-yl)-4-((5R)-2-methyl-2-oxo-1,3,7-triazaspiro-2-phosphaspiro[4.5]decane-7-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[4,3-d]pyrimidine (91 mg). LC/MS(ESI): m/z =697[M+H] ⁺ .

Step D: 6-Fluoro-7-(3-methoxy-7-fluoro-8-ethylisoquinolin-1-yl)-4-((5R)-2-methyl-2-oxo-1,3,7-triazaspiro-2-phosphaspiro[4.5]decan-7-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)pyridin[4,3-d]pyrimidine (69.7 mg, 0.1 mmol) was dissolved in DCM (2 mL) at -78°C, BBr ₃ ((0.4 mL, 1M in DCM, 0.4 mmol) was added, and the mixture was heated to room temperature and stirred for 5 h, then cooled to -78°C and heated with 1N The solution was adjusted to Ph = 7 with NaOH, extracted with EA, and then concentrated by filtration under reduced pressure and purified by preparative HPLC to obtain a light yellow solid compound 66 (28 mg). LC/MS (ESI): m/z = 683 [M + H] ⁺ .

Embodiment 67 Compound 67 (28 mg) was obtained in a similar manner to Example 66. LC/MS (ESI): m/z = 690 [M+H] ⁺ .

Embodiment 68 Compound 68 (32 mg) was obtained by a method similar to Example 66. LC/MS (ESI): m/z = 697 [M+H] ⁺ .

Embodiment 69 Compound 69 (14 mg) was obtained in a similar manner to Example 66. LC/MS (ESI): m/z = 695 [M+H] ⁺ .

Embodiment 70

Step A: Mix 2,4,7-trichloro-8-fluoropyrido[4,3-d]pyrimidine (2.52 g, 10 mmol), N,N-dimethyl-5,6,7,8-tetrahydro-4H-pyrazolo[1,5-a][1,4]diazepine-2-carboxamide (2.29 g, 11 mmol), potassium carbonate (2.07 g, 15 mmol), a catalytic amount of potassium iodide and DMF (60 mL), heat to 120°C, and stir for 4 hours. The mixture was cooled to room temperature and evaporated under reduced pressure to obtain yellow solid 5-(2,7-dichloro-6-fluoropyrido[4,3-d]pyrimidin-4-yl)-N,N-dimethyl-5,6,7,8-tetrahydro-4H-pyrazolo[1,5-a][1,4]diazepine-2-carboxamide (3.61 g). LC/MS(ESI): m/z =424[M+H] ⁺ .

Step B: 5-(2,7-dichloro-6-fluoropyrido[4,3-d]pyrimidin-4-yl)-N,N-dimethyl-5,6,7,8-tetrahydro-4H-pyrazolo[1,5-a][1,4]diazepine-2-carboxamide (424 mg, 1 mmol), (2R,8S)-2-fluoro-1,2,3,5,6,7-hexahydropyrrolizin-7-yl]methanol (175 mg, 1.1 mmol), potassium carbonate (0.18 mg, 2.1 mmol) a catalytic amount of potassium iodide and DMF (8 mL) were mixed, heated to 120°C, and stirred for 4 hours. The mixture was cooled to room temperature and evaporated under reduced pressure to give yellow solid 5-(2(((2R,7aS)-2-fluorotetrahydro-1H-pyridine-7a(5H)-yl)methoxy)-7-chloro-6-fluoropyrido[4,3-d]pyrimidin-4-yl)-N,N-dimethyl-5,6,7,8-tetrahydro-4H-pyrazolo[1,5-a][1,4]diazepine-2-carboxamide (437 mg). LC/MS(ESI): m/z =548[M+H] ⁺ .

Step C: 5-(2(((2R,7aS)-2-fluorotetrahydro-1H-pyrinylcyclo-7a(5H)-yl)methoxy)-7-chloro-6-fluoropyrido[4,3-d]pyrimidin-4-yl)-N,N-dimethyl-5,6,7,8-tetrahydro-4H-pyrazolo[1,5-a][1,4]diazide-2-carboxamide (109.6 mg, 0.2 mmol), 3-methoxy-7-fluoro-8-ethylisoquinolinylboronic acid pinacol ester (66.4 mg, 0.2 mmol), methanesulfonic acid [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) (16 mg, 0.022 mmol), K ₃ PO ₄ , THF (5 mL) and water (1 mL) were mixed and then heated to 60°C under reflux and stirred for 16 hours. The reaction was cooled to room temperature and stirred overnight to obtain a light yellow precipitate. The reaction mixture was diluted with water (2 mL) and the solid was collected by filtration. The crude product was purified by column chromatography to obtain 5-(2(((2R,7aS)-2-fluorotetrahydro-1H-pyridine-7a(5H)-yl)methoxy)-7-(3-methoxy-7-fluoro-8-ethylisoquinolin-1-yl)-6-fluoropyrido[4,3-d]pyrimidin-4-yl)-N,N-dimethyl-5,6,7,8-tetrahydro-4H-pyrazolo[1,5-a][1,4]diazepine-2-carboxamide (94 mg). LC/MS(ESI): m/z =716[M+H] ⁺ .

Step D: 5-(2(((2R,7aS)-2-fluorotetrahydro-1H-pyridine-7a(5H)-yl)methoxy)-7-(3-methoxy-7-fluoro-8-ethylisoquinolin-1-yl)-6-fluoropyrido[4,3-d]pyrimidin-4-yl)-N,N-dimethyl-5,6,7,8-tetrahydro-4H-pyrazolo[1,5-a][1,4]diazepine-2-carboxamide (71.6 mg, 0.1 mmol) was dissolved in DCM (2 mL) at -78°C, BBr ₃ ((0.4 mL, 1M in DCM, 0.4 mmol) was added, and the mixture was heated to room temperature and stirred for 5 h, then cooled to -78°C and heated with 1N The solution was adjusted to Ph = 7 with NaOH, extracted with EA, and then concentrated by filtration under reduced pressure and purified by preparative HPLC to obtain a light yellow solid compound 66 (32 mg). LC/MS (ESI): m/z = 702 [M + H] ⁺ .

Embodiment 71 Compound 71 (25 mg) was obtained in a similar manner to Example 70. LC/MS (ESI): m/z = 709 [M+H] ⁺ .

Embodiment 72 Compound 72 (29 mg) was obtained in a similar manner to Example 70. LC/MS (ESI): m/z = 684 [M+H] ⁺ .

Embodiment 73 Compound 73 (34 mg) was obtained in a similar manner to Example 70. LC/MS (ESI): m/z = 691 [M+H] ⁺ .

Embodiment 74 Compound 74 (28 mg) was obtained in a similar manner to Example 70. LC/MS (ESI): m/z = 702 [M+H] ⁺ .

Embodiment 75 Compound 75 (24 mg) was obtained in a similar manner to Example 70. LC/MS (ESI): m/z = 709 [M+H] ⁺ .

Embodiment 76 Compound 71 (15 mg) was obtained in a similar manner to Example 70. LC/MS (ESI): m/z = 719 [M+H] ⁺ .

Embodiment 75 Compound 75 (18 mg) was obtained in a similar manner to Example 70. LC/MS (ESI): m/z = 726 [M+H] ⁺ .

Embodiment 76 Compound 76 (16 mg) was obtained in a similar manner to Example 70. LC/MS (ESI): m/z = 731 [M+H] ⁺ .

Embodiment 77 Compound 77 (13 mg) was obtained in a similar manner to Example 70. LC/MS (ESI): m/z = 733 [M+H] ⁺ .

Embodiment 78 Compound 78 (24 mg) was obtained in a similar manner to Example 70. LC/MS (ESI): m/z = 714 [M+H] ⁺ .

Embodiment 79 Compound 71 (19 mg) was obtained in a similar manner to Example 70. LC/MS (ESI): m/z = 716 [M+H] ⁺ .

Embodiment 80 Compound 80 (15 mg) was obtained in a similar manner to Example 70. LC/MS (ESI): m/z = 696 [M+H] ⁺ .

Embodiment 81 Compound 81 (14 mg) was obtained in a similar manner to Example 70. LC/MS (ESI): m/z = 698 [M+H] ⁺ .

Embodiment 82 Compound 82 (18 mg) was obtained in a similar manner to Example 70. LC/MS (ESI): m/z = 714 [M+H] ⁺ .

Embodiment 83 Compound 83 (19 mg) was obtained in a similar manner to Example 70. LC/MS (ESI): m/z = 716 [M+H] ⁺ .

Embodiment 84

Step A: Mix 7-bromo-2,4-dichloro-6,8-difluoroquinazoline (3.13 g, 10 mmol), 3-azabicyclo[3.2.1]octane-6-ol hydrochloride (1.79 g, 11 mmol), potassium carbonate (2.07 g, 15 mmol), catalytic amount of potassium iodide and DMF (60 mL), heat to 120°C, and stir for 4 hours. Cool to room temperature, evaporate under reduced pressure to obtain yellow solid 7-bromo-2-chloro-4-(6-hydroxy-3-azabicyclo[3.2.1]octane-3-yl)-6,8-difluoroquinazoline (2.82 g). LC/MS(ESI): m/z =343[M+H] ⁺ .

Step B: 7-Bromo-2-chloro-4-(6-hydroxy-3-azabicyclo[3.2.1]octan-3-yl)-6,8-difluoroquinazoline (467 mg, 1 mmol), (2R,8S)-2-fluoro-1,2,3,5,6,7-hexahydropyrrolizin-7-yl]methanol (175 mg, 1.1 mmol), potassium carbonate (0.18 mg, 2.1 mmol) a catalytic amount of potassium iodide and DMF (8 mL) were mixed, heated to 120°C, and stirred for 4 hours. The mixture was cooled to room temperature and evaporated under reduced pressure to give yellow solid 7-bromo-6,8-difluoro-2(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-(6-hydroxy-3-azabicyclo[3.2.1]octan-3-yl)quinazoline (342 mg). LC/MS(ESI): m/z =466[M+H] ⁺ .

Step C: 7-Bromo-6,8-difluoro-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)-4-(6-hydroxy-3-azabicyclo[3.2.1]octan-3-yl)quinazoline (93 mg, 0.2 mmol), 3-methoxy-7-fluoro-8-ethylisoquinolinylboronic acid pinacol ester (66.4 mg, 0.2 mmol), [n-butyldi(1-adamantyl)phosphine](2-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (16 mg, 0.022 mmol), K ₃ PO ₄ , THF (5 mL) and water (1 mL) were added to the mixture. mL) and then refluxed to 60°C and stirred for 16 hours. The reaction mixture was cooled to room temperature and stirred overnight to obtain a light yellow precipitate. The reaction mixture was diluted with water (2 mL) and the solid was collected by filtration. The crude product was column chromatographed to obtain 6,8-difluoro-7-(3-methoxy-7-fluoro-8-ethylisoquinolin-1-yl)-4-(6-hydroxy-3-azabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)quinazoline (88 mg). LC/MS(ESI): m/z =635[M+H] ⁺ .

Step D: 6,8-difluoro-7-(3-methoxy-7-fluoro-8-ethylisoquinolin-1-yl)-4-(6-hydroxy-3-azabicyclo[3.2.1]octan-3-yl)-2-(((2R,7aS)-2-fluorotetrahydro-1H-pyrin-7a(5H)-yl)methoxy)quinazoline (63.5 mg, 0.1 mmol) was dissolved in DCM (2 mL) at -78°C, BBr ₃ ((0.4 mL, 1M in DCM, 0.4 mmol) was added, and then the mixture was heated to room temperature and stirred for 5 h, then cooled to -78°C and heated with 1N The pH was adjusted to 7 with NaOH, extracted with EA, and then concentrated by filtration under reduced pressure and purified by preparative HPLC to obtain a light yellow solid compound 84 (24 mg). LC/MS (ESI): m/z = 621 [M+H] ⁺ .

Embodiment 85 Compound 85 (24 mg) was obtained in a similar manner to Example 84. LC/MS (ESI): m/z = 621 [M+H] ⁺ .

Embodiment 86 Compound 86 (22 mg) was obtained by a method similar to Example 84. LC/MS (ESI): m/z = 628 [M+H] ⁺ .

Embodiment 87 Compound 87 (19 mg) was obtained in a similar manner to Example 84. LC/MS (ESI): m/z = 628 [M+H] ⁺ .

Embodiment 88 Compound 88 (22 mg) was obtained in a similar manner to Example 17. LC/MS (ESI): m/z = 584 [M+H] ⁺ .

Embodiment 89 Compound 86 (22 mg) was obtained in a similar manner to Example 25. LC/MS (ESI): m/z = 583 [M+H] ⁺ .

Embodiment 90 Compound 86 (22 mg) was obtained in a similar manner to Example 17. LC/MS (ESI): m/z = 602 [M+H] ⁺ .

Embodiment 91 Compound 86 (22 mg) was obtained in a similar manner to Example 25. LC/MS (ESI): m/z = 601 [M+H] ⁺ .

Embodiment 92 Compound 86 (22 mg) was obtained in a similar manner to Example 13. LC/MS (ESI): m/z = 602 [M+H] ⁺ .

Embodiment 93 Compound 86 (22 mg) was obtained in a similar manner to Example 1. LC/MS (ESI): m/z = 584 [M+H] ⁺ .

Embodiment 94 Compound 86 (22 mg) was obtained in a manner similar to Example 13. LC/MS (ESI): m/z = 601 [M+H] ⁺ .

Embodiment 95 Compound 86 (22 mg) was obtained by a method similar to Example 13. LC/MS (ESI): m/z = 683 [M+H] ⁺ . The following compounds were obtained by synthesizing in a manner similar to Examples 88-95 :

Embodiment 96 Biological activity test

Experimental example 1. KRAS Inhibitory activity test

1. Experimental purpose:

Using the TR-FRET method, we screened out compounds that can effectively inhibit the binding of KRas<WT>, KRas<G12A>, KRas<G12C>, KRas<G12D>, KRas<G12R>, KRas<G12S>, KRas<G12V>, KRas<G13D>, and KRas<Q61H> to GTP.

2. Reagent preparation:

a. Storage of reagents:

1) KRAS nucleotide exchange buffer Take 20mL 1000mM HEPES, 20mL 500mM EDTA, 10mL 5M sodium chloride, 100% 0.1mL Tween 20, 949.9mL water, prepare 1L solution, sterilize by filtration, and store at 4°C.

2) KRAS experimental buffer Take 20mL 1000mM HEPES, 10mL 1000mM magnesium chloride, 30mL 5M sodium chloride, 100% 0.05mL Tween 20, 939.95mL water, prepare 1L solution, sterilize by filtration, and store at 4°C.

3) KRAS/Bodipy GDP/Tb-SA mixed solution Take 9.5μL 95μM KRAS＜G12D＞ protein and mix with 440.5μL KRAS nucleotide exchange buffer. Incubate at room temperature for 1 hour, then mix with 8.4μL 17.9μM Tb-SA, 1.8μL 5mM Bodipy GDP, and 9539.8μL KRAS experimental buffer to make a 1L solution. After mixing, let stand at room temperature for 6 hours and store at -80°C.

b. Experimental reagents:

1) KRAS enzyme solution Take 73.3μL KRAS/Bodipy GDP/Tb-SA mixed solution and 2126.7μL KRAS assay buffer to prepare a 2200μL solution.

2) SOS/GTP mixture

c. Experimental process: Take 1.59μL 166μM SOS protein, 198μL 100mM GTP, 2000.41μL KRAS experimental buffer and prepare a 2200μL solution. The concentration of the control compound stock solution is 1mM, and the concentration of the test compound stock solution is 10mM. Transfer 9 μL of control compound and test compound to 384-LDV plate; use Bravo to dilute the compound on LDV plate 10 times; use ECHO to transfer 9 nL of compound on LDV plate to experimental plate; use Dragonfly automatic sampler to add 3 μL of 3nM Kras/0.5nM TB-SA/30nM BodipyGDP mixture and 3 μL of Ras buffer to each well of the experimental plate in sequence, centrifuge the experimental plate at 1000 rpm/min for 1 minute; incubate the experimental plate at room temperature for 1 hour; use Dragonfly automatic sampler to add 3 μL of 120nM SOS/9mM GTP mixture, centrifuge the test plate at 1000rpm/min for 1 minute; incubate the test plate at room temperature for 1 hour; use Envision to read the plate and record the data; use Excel and Xlfit to analyze the data and calculate the _IC50 of the test compound. "++++" means _IC50 ≤5 nM; "+++" means 5nM＜ _IC50 ≤50 nM; "++" means 50nM＜ _IC50 ≤2000 nM; "+" means 2000nM＜ _IC50 .

Table 1 IC50 values of compounds for KRAS enzyme inhibition. No. WT G12A G12C G12D G12R G12S G12V G13D Q61H 1 ++++ ++++ ++++ +++ ++++ ++++ ++++ ++++ ++++ 2 +++ 3 +++ 4 +++ 5 ++++ ++++ 6 ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ 7 ++++ ++++ 8 ++++ ++++ 9 ++++ ++++ 10 ++++ ++++ 11 ++++ ++++ 12 ++++ ++++ 13 ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ 14 ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ 15 ++++ ++++ 16 ++++ ++++ 17 ++++ ++++ 18 ++++ ++++ 19 ++++ ++++ twenty one ++++ ++++ twenty three ++++ ++++ twenty four ++++ ++++ 33 ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ 34 ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ 35 ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ 36 ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ 37 ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ 38 ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ 39 ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ 40 ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ 51 ++++ ++++ 52 ++++ ++++ 53 ++++ ++++ 54 ++++ ++++ 55 ++++ ++++ 56 ++++ ++++ 57 ++++ ++++ 58 ++++ ++++ 59 ++++ ++++ 60 ++++ ++++ 61 ++++ ++++ 62 ++++ ++++ 63 ++++ ++++ 64 ++++ ++++ 65 ++++ ++++ 66 ++++ ++++ 70 ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ 71 ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ 72 ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ 73 ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ 74 ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ 75 ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ 76 ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ 84 ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ 85 ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ 86 ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ 87 ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++++ 90 ++++ 92 ++++ 93 ++++

Experimental example 2. Cells p-ERK Inhibition test

Using the HTRF method, we screened out compounds that can effectively inhibit p-ERK in AsPC-1 (G12D), A549 (G12S), HCT116 (G13D), NCI-H358 (G12C), NCI-H460 (Q61H), NCI-H727 (G12V), MKN1 (WTdep), and PSN-1 (G12R) cells.

Cells were seeded in a transparent 96-well cell culture plate, with 80 μL of cell suspension per well, and each well contained 8,000 cells. The cell plate was placed in a carbon dioxide incubator and incubated overnight at 37 degrees. 2 μL of compound was added to 78 μL of cell culture medium, and after mixing, 20 μL of compound solution was added to the corresponding well of the cell plate, and the cell plate was returned to the carbon dioxide incubator and incubated for 1 hour. After the incubation, the cell supernatant was discarded and 50 μL of 1X cell lysis buffer was added to each well, and incubated at room temperature for 30 minutes. Phospho-ERK1/2 Eu Cryptate antibody and Phospho-ERK1/2 d2 were added using detection buffer. Dilute the antibody 20 times and take 16 μL of cell lysate supernatant to each well in a new 384-well white microplate. Then add 2 μL of Phospho-ERK1/2 Eu Cryptate antibody dilution and 2 μL of Phospho-ERK1/2 d2 antibody dilution. Incubate at room temperature for at least 4 hours. After the incubation, use a multi-label analyzer to read HTRF excitation: 320nm, emission: 615nm, 665nm;

Calculate the IC ₅₀ of the test compound. "++++" indicates IC ₅₀ ≤10 nM; "+++" indicates 10nM＜IC ₅₀ ≤100 nM; "++" indicates 100nM＜IC ₅₀ ≤2000 nM; "+" indicates 2000nM＜IC ₅₀ .

Table 2. IC ₅₀ (nM) of compounds on inhibition of p-ERK in tumor cells. No. AsPC-1 A549 HCT116 NCI-H358 NCI-H460 NCI-H727 MKN1 PSN-1 1 ++ + 2 ++ + 3 ++ + 4 ++ ++ + 19 ++++ ++++ +++ ++++ ++++ ++++ ++++ ++ twenty three ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++ 31 ++++ ++++ +++ ++++ ++++ ++++ ++++ ++ 33 ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++ 34 ++++ ++++ +++ ++++ ++++ ++++ ++++ ++ 35 ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++ 36 ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++ 37 ++++ ++++ +++ ++++ ++++ ++++ ++++ ++ 38 ++++ ++++ ++++ ++++ ++++ ++++ ++++ ++ 56 ++++ +++ ++++ ++++ ++++ ++++ ++++ ++ 57 ++++ +++ ++++ ++++ ++++ ++++ ++++ ++ 70 ++++ ++ ++ +++ ++ +++ +++ + 71 ++++ ++ ++ +++ ++ +++ +++ + 72 ++++ ++ ++ +++ ++ +++ +++ + 73 ++++ ++ ++ +++ ++ +++ +++ + 74 ++++ ++ ++ +++ ++ +++ +++ + 75 ++++ ++ ++ +++ ++ +++ +++ + 84 ++++ ++++ ++++ ++++ ++++ ++++ +++ 85 ++++ ++++ ++++ ++++ ++++ ++++ +++ 86 ++++ ++++ ++++ ++++ ++++ ++++ +++ 87 ++++ ++++ ++++ ++++ ++++ ++++ +++

Although the present invention has been described in detail above, it is understood by those skilled in the art that various modifications and changes can be made to the present invention without departing from the spirit and scope of the present invention. The scope of the present invention is not limited to the detailed description above, but should belong to the scope of the patent application.

without

without.

without.

Claims (7)

A compound represented by general formula (I), a stereoisomer or a pharmaceutically acceptable salt thereof, (I) each L ₂ is independently selected at each occurrence from a bond, OC _0-6 alkyl, NHC _0-6 alkyl, C _1-6 alkyl, COC _0-6 alkyl or SC _0-6 alkyl; each R ₁ is independently selected at each occurrence from H, D, halogen, C _1-6 alkyl, -C _2-6 alkenyl, -C _2-6 alkynyl, CN, OC _1-6 alkyl; each R ₁ is independently optionally substituted or unsubstituted with 1, 2, 3, 4, 5 or 6 substituents selected from deuterium, halogen, -C _1-6 alkyl; n is 1-3; each X ₁ , X ₂ , X ₃ is independently selected at each occurrence from N, CR ₃ ; each R ₃ is independently selected from H, D, cyano, halogen, C _1-6 alkyl, CN; each R _{R 4} is independently selected from H, D, C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, CN, C _3-6 carbocyclyl, 3-10 membered heterocyclic ring, 4-10 membered heterofused ring; 3-8 membered heterocyclic ring contains 1, 2, 3 or 4 heteroatoms selected from N, O or S at each occurrence; each R ₄ is independently optionally substituted with 1, 2, 3, 4, 5 or 6 substituents selected from deuterium, halogen, C _1-6 alkyl, -C _1-6 alkoxy, oxo, OC _1-6 alkyl, C _3-6 carbocyclyl, 3-10 membered heterocyclic ring or unsubstituted; U is selected from 3-8 membered cycloalkyl, 3-8 membered heterocycloalkyl, 5-12 membered fused alkyl, 5-12 membered fused heterocyclo, 5-12 membered spirocyclo, 5-12 membered spiroheterocyclo, aromatic or heteroaromatic, each heterocycloalkyl, fused heterocyclo, spiroheterocyclo and heteroaromatic independently containing 1, 2, 3 or 4 heteroatoms selected from N, O or S at each occurrence, wherein the cycloalkyl, heterocycloalkyl, spirocyclo, fused cyclo, fused heterocyclo, spiroheterocyclo, aromatic or heteroaromatic is optionally substituted by one or more ^G1 ; ^G1 is independently selected from deuterium, cyano, halogen, _C1-6 alkyl, _C2-6 alkenyl, C _-OR5 , -OC(O)NR5R6, -C(O ₎ OR5, -C(O) _NR5R6 , -C(O _)R5 , -NR5R6, _-NR5C (O) _R6 , _-NR5C (O _{) NR6R7} , _-S ( _O ) _iR5 , _{or -NR5S} (O) _iR6 , wherein _the alkyl, _alkenyl , _alkynyl , _{cycloalkyl ,} heterocycloalkyl, _{aryl ,} heteroaryl is optionally substituted with one or more deuterium, cyano, halogen, _C1-7 alkyl, _C2-7 alkenyl, _C2-7 alkynyl, C R5, R6, R7, R8 _, R9 and R10 are _each independently _selected from hydrogen _, deuterium _, cyano, halogen, _C1-6 alkyl, _C2-6 alkyl, C3-7 alkyl, _C4-7 alkyl, C5-8 alkyl, _C6-7 alkyl, _{C7-10 aryl} , C8-12 aryl _{, C9-10} aryl, C1-6 alkyl, C1-6 alkyl, _{C2-7 alkyl} , C3-7 _alkyl , _C4-7 alkyl, _C5-8 alkyl, _C6-7 alkyl, C7-10 aryl, C8-12 aryl, _C9-10 aryl, _C8-12 alkyl, _C1-6 alkyl _{, C2-7} alkyl, _C3-7 alkyl, C4-7 alkyl, C5-8 alkyl, _C6-7 alkyl, C7-10 _aryl , C8-12 _aryl , _3-8 membered cycloalkyl or 3-8 membered monocyclic heterocyclic group, monocyclic heteroaryl group or phenyl group; and i is 1 or 2. A compound represented by general formula (II), a stereoisomer or a pharmaceutically acceptable salt thereof: (II) each L ₂ is independently selected at each occurrence from a bond, OC _0-6 alkyl, NHC _0-6 alkyl, C _1-6 alkyl, COC _0-6 alkyl or SC _0-6 alkyl; each R ₁ is independently selected at each occurrence from H, D, halogen, C _1-6 alkyl, -C _2-6 alkenyl, -C _2-6 alkynyl, CN, OC _1-6 alkyl; each R ₁ is independently optionally substituted with 1, 2, 3, 4, 5 or 6 substituents selected from deuterium, halogen, -C _1-6 alkyl or unsubstituted; n is 1-3; each X ₁ is independently selected at each occurrence from N, CR ₃ ; each R ₃ is independently selected from H, D, cyano, halogen, C _1-6 alkyl, CN; Y ₁ , Y ₂ , Y ₃ , Y ₄ at most one selected from the heteroatom N, O, S, the others selected from CR ₃ ; each R ₃ is independently selected from H, D, cyano, halogen, C _1-6 alkyl, CN; each R ₄ is independently selected from H, D, C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, CN, C _3-6 carbocyclic group, 3-10 membered heterocyclic ring, 4-10 membered heterofused ring; 3-8 membered heterocyclic ring contains 1, 2, 3 or 4 heteroatoms selected from N, O, or S at each occurrence; each R ₄ is independently optionally substituted by 1, 2, 3, 4, 5 or 6 deuterium, halogen, C _1-6 alkyl, -C _1-6 alkoxy, oxo, OC _1-6 alkyl, C G is selected from _3-8 membered cycloalkyl, 3-8 membered heterocycloalkyl, 5-12 membered fused alkyl, 5-12 membered fused heterocyclo, 5-12 membered spirocyclo, 5-12 membered spiroheterocyclo, aromatic or heteroaromatic, each heterocycloalkyl, fused heterocyclo, spiroheterocyclo and heteroaromatic independently containing 1, 2, 3 or 4 heteroatoms selected from N, O or S at each occurrence, wherein the cycloalkyl, heterocycloalkyl, spirocyclo, fused heterocyclo, fused heterocyclo, spiroheterocyclo, aromatic or heteroaromatic is optionally substituted by one or more G1; G is selected from 3-8 membered cycloalkyl, 3-8 membered heterocycloalkyl, 5-12 membered fused heterocyclo, 5-12 membered spirocyclo, 5-12 membered spiroheterocyclo, aromatic or heteroaromatic, each heterocycloalkyl, fused heterocyclo, spiroheterocyclo and heteroaromatic independently containing 1, 2, 3 or 4 heteroatoms selected from N, O or S at each occurrence, wherein the cycloalkyl, heterocycloalkyl, spirocyclo, fused heterocyclo, fused heterocyclo, spiroheterocyclo, aromatic or heteroaromatic is optionally substituted by one or more ^G1 ; ¹ are each independently selected from deuterium, cyano, halogen, C _1-6 alkyl, C _2-6 alkenyl, C _2-6 alkynyl, C _3-8 cycloalkyl or 3-8 membered heterocyclic group, C _6-10 aryl, 5-10 membered heteroaryl, -OR ₅ , -OC(O)NR ₅ R ₆ , -C(O)OR ₅ , -C(O)NR ₅ R ₆ , -C(O)R ₅ , -NR ₅ R ₆ , -NR ₅ C(O)R ₆ , -NR ₅ C(O)NR ₆ R ₇ , -S(O) _i R ₅ or -NR ₅ S(O) _i R ₆ wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl is optionally substituted with one or more deuterium, cyano, halogen, _C1-7 alkyl, _C2-7 alkenyl, _C2-7 alkynyl, _C3-9 cycloalkyl or 3-9 membered heterocyclo, _C7-10 aryl, 6-10 membered heteroaryl, _-OR8 , -OC(O) _NR8R9 , -C(O) _OR8 , _-C (O) _NR8R9 , -C(O) _R8 , _-NR8R9 , _-NR8C (O _{)R9 , -NR8C} (O) _{NR9R10 , -S} (O _{)iR8 or -NR8S} (O) _iR9 substituents _{; R5} , _R6 R ₇ , R ₈ , R ₉ and R ₁₀ are each independently selected from hydrogen, deuterium, cyano, halogen, C _1-6 alkyl, C _3-8 cycloalkyl or 3-8 membered monocyclic heterocyclic group, monocyclic heteroaromatic group or phenyl; and i is 1 or 2. The compound of general formula (I) and (II) as described in claim 1 or 2, a pharmaceutically acceptable salt thereof or a stereoisomer thereof, which is selected from: . A compound as described in any one of claims 1 to 3, or an optical isomer thereof, a pharmaceutically acceptable salt, a prodrug, a deuterated derivative, a hydrate, a solvate, wherein the pharmaceutically acceptable salt is selected from the following group: potassium salt, sodium salt, magnesium salt, calcium salt, sulfate, hydrochloride, phosphate, sulfonate, or carbonate. A pharmaceutical composition, characterized in that it comprises the compound described in any one of claims 1 to 4, or its optical isomer, a pharmaceutically acceptable salt, a prodrug, a deuterated derivative, a hydrate, a solvate, and a pharmaceutically acceptable carrier. A use of a compound of any one of claims 1 to 4, or an optical isomer, pharmaceutically acceptable salt, prodrug, deuterated derivative, hydrate, or solvate thereof, characterized in that it is used to prepare a pharmaceutical composition for treating a disease, disorder, or condition associated with the activity or expression of KRas, KRas G12A, KRas G12C, KRas G12D, KRas G12R, KRas G12S, KRas G12V, KRas G13D, or KRas Q61H. The use as claimed in claim 6, wherein the disease, disorder or condition is selected from the group consisting of pancreatic cancer, non-small cell lung cancer, small cell lung cancer, lung adenocarcinoma, lung squamous cell carcinoma, colon cancer, colorectal cancer, thyroid cancer, embryonal rhabdomyosarcoma, dermatomyeloma, melanoma, liver cancer, rectal cancer, bladder cancer, pharyngeal cancer, breast cancer, prostate cancer, Prostate cancer, neuroglioma, ovarian cancer, head and neck squamous cell carcinoma, cervical cancer, esophageal cancer, kidney cancer, skin cancer, lymphoma, gastric cancer, acute myeloid leukemia, myelofibrosis, B-cell lymphoma, monocytic leukemia, splenomegaly, multiple eosinophilia syndrome, bone marrow cancer and other solid tumors and blood tumors.